TWI768465B - Tetrahydroindazole derivatives and their preparation - Google Patents

Abstract

The invention discloses a tetrahydroindazole compound represented by formula (I) and its stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides, prodrugs and pharmaceutical compositions thereof, Preparation method, and use in the prevention and treatment of diseases mediated by cannabinoid CB2 receptors.

Description

Tetrahydroindazole derivatives and their preparation

The present invention relates to a tetrahydroindazole compound and its stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides, prodrugs and their pharmaceutical compositions, preparation methods, as well as in the preparation of cannabis Use in the prevention and treatment of CB2 receptor-mediated diseases.

Neuropathic pain results from damage to peripheral or central nerve pathways, resulting in persistent spontaneous pain and hypersensitivity to pain and innocuous stimuli. The underlying etiologies of neuropathic pain vary widely but generally have similar clinical features. Some of the most common pathologies that cause neuropathic pain are diabetic neuropathy, amputation, surgery, and postherpetic neuralgia. Studies have shown that up to 7-8% of adults suffer from neuropathic pain, and the prevalence is increasing and is expected to increase further in the aging population. In addition to personal distress, increased treatment costs due to unemployment and social incapacity are often accompanied by complications such as anxiety and depression.

The main symptoms of Crohn's disease include abdominal pain, diarrhea, and fatigue, weight loss, fever, growth retardation, anemia, and recurrent anal fistulas or other extraintestinal manifestations. Drug therapy includes aminosalicylic acid preparations, glucocorticoids, immunosuppressants, antibiotics and anti-TNF-α monoclonal antibodies. There are currently no medications to manage abdominal pain associated with Crohn's disease.

Pain is one of the most common clinical symptoms and one of the main factors for patients to seek medical services. Currently, the widely used analgesics on the market can cause harmful side effects and unsatisfactory treatment effects. The development of new painkillers is urgent.

Cannabis extracts have been used for pain relief for centuries. In addition to Delta-9-THC, cannabidiol (CBD), there are a variety of endogenous ligand substances, as well as chemically synthesized cannabinoids (cannabinoid), exerts analgesic and euphoric effects by binding to cannabinoid receptors. And cannabinoids are a class of extracellular signaling molecules found in both plants and animals. Signaling from these molecules is mediated in animals through two G protein-coupled receptors, cannabinoid receptor 1 (CBl) and cannabinoid receptor 2 (CB2). CB1 is most abundantly expressed in neurons of the CNS, but is also present at lower concentrations in various peripheral tissues and cells (Matsuda, L.A. et al. (1990) Nature 346:561-564). In contrast, CB2 is predominantly, but not exclusively, expressed in non-neural tissues such as hematopoietic cells, endothelial cells, osteoblasts, osteoclasts, endocrine pancreas, and cancer cell lines (Munro, S. et al. (1993) Nature 365 : 61-65; and Pache, P. et al. (2006) Pharmacol. Rev. 58(3): 389-462). Central CB1 is distributed in the cerebral cortex and limbic system and is responsible for exerting analgesic effects and causing behavioral changes after cannabinoid priming. Although CB1 can mediate potent analgesic effects, it can cause psychotic symptoms such as euphoria, ataxia, and dizziness, and may produce addiction and tolerance, which limits its use in the field of analgesia. Peripheral CB2 is mainly distributed in immune cells and plays a role in pain and inflammatory signaling. In addition, CB2 is also distributed in peripheral nerve fibers and nociceptive nerve endings. After CB2 activation, it exerts analgesic effect by inhibiting the production of toxins and inflammatory mediators by neutrophils and macrophages, and can also block the excitation conduction of injured nerves. CB2 is expressed in the brainstem, cerebral cortex and cerebellum of mice, but the expression is very low, about 3.4% of the spleen.

Peripheral selective CB2 agonists can theoretically avoid CB1R-related central psychiatric side effects and show good analgesic effects in multiple preclinical models. Several CB2 selective agonists were discontinued in the analgesic field due to insufficient analgesic efficacy in clinical trials, including JBT-101, LY2828360, etc. The reasons may be as follows: Although it exhibits a certain selectivity, it still has an agonistic effect on CB1, so it may lead to adverse events at effective doses, and the drug effect is insufficient at a safe dose; the agonistic effect on CB1 may lead to significant drug effects in animal models. Insufficient drug effect after transformation. CB2 agonists will have functional selectivity. The downstream of CB1/2 includes adenosyl cyclase synthesis, ERK signaling pathway activation, ion current changes, internalization, β-arrestin, etc. Different drugs have functional biases for downstream activation. .

The purpose of the present invention is to introduce a new class of cannabinoid CB2 agonists that are effective, safe, highly selective, long-acting, and have good pharmacokinetic characteristics.

The present invention relates to a tetrahydroindazole compound represented by formula (I) and a stereoisomer, pharmaceutically acceptable salt, solvate, hydrate, N-oxide or prodrug thereof:

、

或者

。 Wherein, R ¹ is selected from -L ₁ -L ₂ -L ₃ -L ₄ ; L ₁ is selected from C _1-6 alkylene, C _3-7 cycloalkenylene, C _3-7 cycloalkylene, C _3-6 unsaturated cycloalkylene group, C _3-6 heterocyclic group, C _6-10 arylene group, C _5-10 heteroarylene group, C _4-12 bridged ring, C _4-12 parallel ring, C _5-10 spiro rings or absent, the alkylene, cycloalkenylene, cycloalkylene, unsaturated cycloalkylene, heterocyclylene, arylene, heteroarylene, bridged ring, and The ring and spiro ring are each independently optionally substituted by the following substituents: halogen, cyano, hydroxyl, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, -C _2-6 alkynyl- C _3-6 cycloalkyl, C _1-6 alkoxy, hydroxy-substituted C _1-6 alkyl, halogenated C _1-6 alkyl, benzene ring, -C _1-6 alkyl-O-haloalkyl , -C(O)OC _1-6 alkyl or C _3-6 cycloalkyl; L ₂ is selected from C _1-6 alkylene, C _3-7 cycloalkylene, C _3-6 heterocyclylene , C _5-10 heteroarylene, -C(O)NH-, -C(O)-, -C(O)O- or absent, the alkylene, cycloalkylene, hetero The cyclic group and the heteroarylene group are each independently optionally substituted by the following substituents: C _1-6 alkyl or halogen; L ₃ is selected from C _1-6 alkylene or absent; L ₄ is selected from H, halogen, hydroxyl , amine group, ureido group, cyano group, C _1-6 alkyl group, C _3-7 cycloalkyl group, C _1-6 alkoxy group, C _3-6 heterocyclic group, C _2-6 alkynyl group, C _{6 -10} aryl, C _5-10 heteroaryl, -NHC _1-6 alkyl, -N(C _1-6 alkyl)C _1-6 alkyl, -NHC(O)C _1-6 alkyl, -C(O)NHC _1-6 alkyl, -NH-C _6-10 aryl, -NH-C _3-7 cycloalkyl, -C(O)-C _6-10 aryl, -OC _{6- 10} aryl, -OC _6-10 heteroaryl, -OC _1-6 alkylene-COOH, -C(O)NH ₂ , -COOH, C _4-12 bridged ring, C _4-12 cyclic ring or C _5-10 Spiro, the alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkynyl, bridged, spiro, and parallel are optionally substituted by the following substituents: halogen, cyano, hydroxyl, amino, C _1-6 alkyl, C _1-6 haloalkyl or C _1-6 alkoxy; R ² and R ³ are each independently selected from H, F, Cl, cyano, Hydroxyl or C _1-6 alkyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H, F, Cl, amino, cyano, hydroxyl, C _1-6 alkyl or C _1-6 haloalkane R ⁸ , R ⁹ are independently selected from H, F, Cl, C _1-6 alkyl, and the alkyl is optionally substituted by the following substituents: cyano, F or Cl, or R ⁸ and R ⁹ together to form C _2-6 alkenyl or C _3-6 Cycloalkyl; R ¹⁰ is selected from H or C _1-6 alkyl; R ¹¹ is selected from aryl or heteroaryl, and the aryl or heteroaryl is optionally substituted by the following substituents: F, Cl, C _1-6 alkyl or C _1-6 alkoxy, the heteroaryl group contains heteroatoms selected from N, O, S and their oxidation states; the condition is: when R ¹¹ is selected from aryl and substituted aryl , R ¹ is selected from

,

or

.

The present invention relates to a tetrahydroindazole compound represented by formula (I) and a stereoisomer, pharmaceutically acceptable salt, solvate, hydrate, N-oxide or prodrug thereof:

、

、

或者

。 Wherein, R ¹ is selected from -L ₁ -L ₂ -L ₃ -L ₄ ; L ₁ is selected from C _1-6 alkylene, C _3-7 cycloalkenylene, C _3-7 cycloalkylene, C _3-6 unsaturated cycloalkylene group, C _3-6 heterocyclic group, C _6-10 arylene group, C _5-10 heteroarylene group, C _4-12 bridged ring, C _4-12 parallel ring, C _5-10 spiro rings or absent, the alkylene, cycloalkenylene, cycloalkylene, unsaturated cycloalkylene, heterocyclylene, arylene, heteroarylene, bridged ring, and The ring and spiro ring are each independently optionally substituted with the following substituents: halogen, cyano, hydroxyl, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _2-6 alkynyl-C _3-6 cycloalkyl, C _1-6 alkoxy, hydroxy-substituted C _1-6 alkyl, halogenated C _1-6 alkyl, benzene ring, C _1-6 alkyl-O-haloalkyl, - C(O)OC _1-6 alkyl or C _3-6 cycloalkyl; L ₂ is selected from C _1-6 alkylene, C _3-7 cycloalkylene, C _3-6 heterocyclylene, C _5-10 Heteroarylene, -C(O)NH-, -C(O)-, -C(O)O- or absent, the alkylene, cycloalkylene, heterocyclylene , Heteroarylene are each independently optionally substituted by the following substituents: C _1-6 alkyl or halogen; L ₃ is selected from C _1-6 alkylene or absent; L ₄ is selected from H, halogen, hydroxyl, amine radical, ureido, cyano, C _1-6 alkyl, C _3-7 cycloalkyl, C _1-6 alkoxy, C _3-6 heterocyclyl, C _1-6 haloalkyl, C _2-6 Alkynyl, C _6-10 aryl, C _5-10 heteroaryl, -NHC _1-6 alkyl, -N(C _1-6 alkyl)C _1-6 alkyl, -NHC(O)C _{1 -6} alkyl, -C(O)NHC _1-6 alkyl, -NH-C _6-10 aryl, -NH-C _3-7 cycloalkyl, -C(O)-C _6-10 aryl , -OC _6-10 aryl, -OC _6-10 heteroaryl, -OC _1-6 alkyl-COOH, -C(O)NH ₂ , -COOH, C _4-12 bridged ring, C _4-12 Paracyclic or _C5-10 spiro, the alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkynyl, bridged, spiro, and paracyclic are optionally selected as follows Substituent substitution: halogen, cyano, hydroxyl, amino, C _1-6 alkyl, C _1-6 haloalkyl or C _1-6 alkoxy; R ² , R ³ are each independently selected from H, F, Cl , cyano, hydroxyl or C _1-6 alkyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H, F, Cl, amino, cyano, hydroxyl, C _1-6 alkyl or C _1-6 haloalkyl; R ⁸ , R ⁹ are each independently selected from H, F, Cl, C _1-6 alkyl, and the alkyl is optionally substituted by the following substituents: cyano, F or Cl, or R ⁸ together with R ⁹ form C _2-6 alkenyl Or C _3-6 cycloalkyl; R ¹⁰ is selected from H or C _1-6 alkyl; R ¹¹ is selected from aryl or heteroaryl, and the aryl or heteroaryl is optionally substituted by the following substituents: F, Cl, C _1-6 alkyl or C _1-6 alkoxy, the heteroaryl group contains heteroatoms selected from N, O, S and their oxidation states; the condition is: when R ¹¹ is selected from aryl and substituted aryl, R ¹ is selected from

,

,

or

.

The present invention relates to a tetrahydroindazole compound represented by formula (I) and a stereoisomer, pharmaceutically acceptable salt, solvate, hydrate, N-oxide or prodrug thereof:

、

、

、吲哚基、吡咯基、吡啶基、呱啶基、苯基、環己基、環戊基、吡嗪、金剛烷基、呱嗪、嗎啉基、

或者

；在某些實施方案中，R¹選自

、

、

、吲哚基、吡咯基、吡啶基、呱啶基、苯基、環己基、環戊基、吡嗪、金剛烷基、呱嗪、嗎啉基、

；在某些實施方案中，R¹選自

、

、

、吲哚基、吡咯基、吡啶基、呱啶基、苯基、環己基、環戊基、吡嗪、金剛烷基、呱嗪、嗎啉基、

；在某些實施方案中，R¹選自

、

、

、

；某些實施方案中，R¹選自

；在某些實施方案中，R¹選自

、

、

、金剛烷基、

、

、

；在某些實施方案中，R¹選自

、

、

、

、

、

；在某些實施方案中，R¹選自

、

、

；在某些實施方案中，R¹選自

；在 某些實施方案中，R¹選自

；R²、R³各自獨立選自H、F、Cl、氰基、羥基、C_1-6烷氧基或者C_1-6烷基；在某些實施方案中，R²、R³各自獨立選自H、F、Cl、氰基、羥基或者C_1-6烷基；在某些實施方案中，R²、R³各自獨立選自H、F、Cl、甲基、乙基、異丙基、叔丁基；在某些實施方案中，R²、R³各自獨立選自H、F或者甲基；R⁴、R⁵、R⁶、R⁷各自獨立選自H、F、Cl、胺基、氰基、羥基、C_1-6烷基或者C_1-6鹵代烷基；在某些實施方案中，R⁴、R⁵、R⁶、R⁷各自獨立選自H、F、氰基、羥基或者C_1-6烷基；R⁴、R⁵、R⁶、R⁷各自獨立選自H或者C_1-6烷基；在某些實施方案中，R⁴、R⁵、R⁶、R⁷各自獨立選自H、甲基、乙基、異丙基；R⁸、R⁹各自獨立選自H、F、Cl、氰基、羥基、C_1-6烷氧基或者C_1-6烷基；R⁸、R⁹各自獨立選自H、F、Cl、C_1-6烷基，所述的烷基任選被以下取代基取代：氰基、F或者Cl，或者R⁸與R⁹一起形成C_2-6烯基或者C_3-6環烷基；R¹⁰選自H或者C_1-6烷基；R¹¹選自芳基或雜芳基，所述的芳基或雜芳基任選被以下取代基取代：F、Cl、C_1-6烷基或者C_1-6烷氧基；在某些實施方案中R¹¹選自

或者

；在某些實施方案中，R¹¹選自

；條件是：當R¹¹選自芳基及取代芳基時，R¹選自

、

或者

。 R ¹ is selected from C _1-6 alkyl, C _3-6 cycloalkyl, C _3-6 heterocycloalkyl, C _3-6 unsaturated cyclic hydrocarbon, C _6-10 aryl, C _5-10 heteroaryl base, C _4-12 bridged ring, C _{4-12 unbroken} ring, C _5-10 spiro ring, the alkyl group, cycloalkyl group, unsaturated cyclic hydrocarbon group, heterocycloalkyl group, aryl group, heteroaryl group, The bridged ring, paracyclic or spiro ring is optionally substituted with the following substituents: F, Cl, hydroxy, cyano, C _1-6 alkyl, hydroxy substituted C _1-6 alkyl, C _1-6 alkoxy, Halogenated C _1-6 alkyl, benzene ring, -C(O)OC _1-6 alkyl, -C _1-6 alkyl-OC _1-6 alkyl or C _3-6 cycloalkyl; R ¹ selected From C _1-6 alkyl, C _3-6 cycloalkyl, C _3-6 heterocycloalkyl, C _3-6 unsaturated cyclic hydrocarbon, C _6-10 aryl, C _5-10 heteroaryl, C _4-12 bridged ring, C _4-12 heterocyclic ring, C _5-10 spiro ring, the alkyl group, cycloalkyl group, unsaturated cyclic hydrocarbon group, heterocycloalkyl group, aryl group, heteroaryl group, bridged ring, The acyl or spiro ring is optionally substituted by the following substituents: F, Cl, hydroxy, cyano, C _1-6 alkyl, hydroxy substituted C _1-6 alkyl, C _1-6 alkoxy, benzene ring, -C(O)OC _1-6 alkyl or C _3-6 cycloalkyl; in certain embodiments, R ¹ is selected from

,

,

, indolyl, pyrrolyl, pyridyl, oxidyl, phenyl, cyclohexyl, cyclopentyl, pyrazine, adamantyl, oxazine, morpholinyl,

or

; In certain embodiments, R ¹ is selected from

,

,

, indolyl, pyrrolyl, pyridyl, oxidyl, phenyl, cyclohexyl, cyclopentyl, pyrazine, adamantyl, oxazine, morpholinyl,

; In certain embodiments, R ¹ is selected from

,

,

, indolyl, pyrrolyl, pyridyl, oxidyl, phenyl, cyclohexyl, cyclopentyl, pyrazine, adamantyl, oxazine, morpholinyl,

; In certain embodiments, R ¹ is selected from

,

,

,

; In certain embodiments, R ¹ is selected from

; In certain embodiments, R ¹ is selected from

,

,

, adamantyl,

,

,

; In certain embodiments, R ¹ is selected from

,

,

,

,

,

; In certain embodiments, R ¹ is selected from

,

,

; In certain embodiments, R ¹ is selected from

; In certain embodiments, R ¹ is selected from

; R ² , R ³ are each independently selected from H, F, Cl, cyano, hydroxyl, C _1-6 alkoxy or C _1-6 alkyl; In certain embodiments, R ² , R ³ are each independently is selected from H, F, Cl, cyano, hydroxyl or C _1-6 alkyl; in certain embodiments, R ² , R ³ are each independently selected from H, F, Cl, methyl, ethyl, isopropyl In certain embodiments, R ² , R ³ are each independently selected from H, F or methyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H, F, Cl, amino, cyano, hydroxyl, C _1-6 alkyl or C _1-6 haloalkyl; in certain embodiments, R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H, F, cyano , hydroxy or C _1-6 alkyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H or C _1-6 alkyl; in certain embodiments, R ⁴ , R ⁵ , R ⁶ , R ⁷ is each independently selected from H, methyl, ethyl, and isopropyl; R ⁸ and R ⁹ are each independently selected from H, F, Cl, cyano, hydroxyl, C _1-6 alkoxy or C _1-6 Alkyl; R ⁸ , R ⁹ are each independently selected from H, F, Cl, C _1-6 alkyl, and the alkyl is optionally substituted by the following substituents: cyano, F or Cl, or R ⁸ and R ⁹ together form C _2-6 alkenyl or C _3-6 cycloalkyl; R ¹⁰ is selected from H or C _1-6 alkyl; R ¹¹ is selected from aryl or heteroaryl, said aryl or heteroaryl The group is optionally substituted with the following substituents: F, Cl, C _1-6 alkyl or C _1-6 alkoxy; in certain embodiments R ¹¹ is selected from

or

; In certain embodiments, R ¹¹ is selected from

; Condition is: when R ¹¹ is selected from aryl and substituted aryl, R ¹ is selected from

,

or

.

The present invention relates to a tetrahydroindazole compound represented by formula (I) and a stereoisomer, pharmaceutically acceptable salt, solvate, hydrate, N-oxide or prodrug thereof:

、

、

、吲哚基、吡咯基、吡啶基、呱啶基、苯基、環己基、環戊基、吡嗪、金剛烷基、呱嗪、嗎啉基、

、

、

、

、

或者

；在某些實施方案中，R¹選自

、

、

、吲哚基、吡咯基、吡啶基、呱啶基、苯基、環己基、環戊基、吡嗪、金剛烷基、呱嗪、嗎啉基、

；在某些實施方案中，R¹選自

、

、

、

；在某些實施方案中，R¹選自

、

、

、金剛烷基、

、

、

；在某些實施方案中，R¹選自

、

、

、

、

、

；在某些實施方案中，R¹選自

、

、

、

；在某些實施方案中，R¹選自

、

、

；在某些實施方案中，R¹選自

；在某些實施方案中，R¹選自

；在某些實施方案中，R¹選自

；R²、R³各自獨立選自H、F、Cl、氰基、羥基、C_1-6烷氧基或者C_1-6烷基； 在某些實施方案中，R²、R³各自獨立選自H、F、Cl、氰基、羥基或者C_1-6烷基；在某些實施方案中，R²、R³各自獨立選自H、F、Cl、甲基、乙基、異丙基、叔丁基；在某些實施方案中，R²、R³各自獨立選自H、F或者甲基；R⁴、R⁵、R⁶、R⁷各自獨立選自H、F、Cl、胺基、氰基、羥基、C_1-6烷基或者C_1-6鹵代烷基；在某些實施方案中，R⁴、R⁵、R⁶、R⁷各自獨立選自H、F、氰基、羥基或者C_1-6烷基；R⁴、R⁵、R⁶、R⁷各自獨立選自H或者C_1-6烷基；在某些實施方案中，R⁴、R⁵、R⁶、R⁷各自獨立選自H、甲基、乙基、異丙基；R⁸、R⁹各自獨立選自H、F、Cl、氰基、羥基、C_1-6烷氧基或者C_1-6烷基；R⁸、R⁹各自獨立選自H、F、Cl、C_1-6烷基，所述的烷基任選被以下取代基取代：氰基、F或者Cl，或者R⁸與R⁹一起形成C_2-6烯基或者C_3-6環烷基；R¹⁰選自H或者C_1-6烷基；R¹¹選自芳基或雜芳基，所述的芳基或雜芳基任選被以下取代基取代：F、Cl、C_1-6烷基或者C_1-6烷氧基；在某些實施方案中R¹¹選自

或者

；在某些實施方案中，R¹¹選自

；條件是：當R¹¹選自芳基及取代芳基時，R¹選自

、

、

或者

。 R ¹ is selected from C _1-6 alkyl, C _3-6 cycloalkyl, C _3-6 heterocycloalkyl, C _3-6 unsaturated cyclic hydrocarbon, C _6-10 aryl, C _5-10 heteroaryl base, C _4-12 bridged ring, C _{4-12 unbroken} ring, C _5-10 spiro ring, the alkyl group, cycloalkyl group, unsaturated cyclic hydrocarbon group, heterocycloalkyl group, aryl group, heteroaryl group, The bridged ring, paracyclic or spiro ring is optionally substituted with the following substituents: F, Cl, hydroxy, cyano, C _1-6 alkyl, hydroxy substituted C _1-6 alkyl, C _1-6 alkoxy, C _3-6 heterocycloalkyl, benzene ring, -C(O)OC _1-6 alkyl, -C _1-6 alkyl-OC _1-6 alkyl or C _3-6 cycloalkyl; in some In an embodiment, R ¹ is selected from the group consisting of C _1-6 alkyl, C _3-6 cycloalkyl, C _3-6 heterocycloalkyl, C _3-6 unsaturated cyclic hydrocarbon, C _6-10 aryl, C _{5 -10} heteroaryl, C _4-12 bridged ring, C _{4-12 unbroken} ring, C _5-10 spiro ring, the alkyl group, cycloalkyl group, unsaturated cyclic hydrocarbon group, heterocycloalkyl group, aryl group, Heteroaryl, bridged, paracyclic or spirocyclic optionally substituted with the following substituents: F, Cl, hydroxy, cyano, C _1-6 alkyl, hydroxy substituted C _1-6 alkyl, C _1-6 alkoxy, benzene ring, -C(O)OC _1-6 alkyl, -C _1-6 alkyl-OC _1-6 alkyl, or C _3-6 cycloalkyl; in certain embodiments, R ¹ is selected from C _1-6 alkyl group, C _3-6 cycloalkyl group, C _3-6 heterocycloalkyl group, C _3-6 unsaturated cyclic hydrocarbon group, C _6-10 aryl group, C _5-10 heteroaryl group , C _4-12 bridged ring, C _{4-12 unbroken} ring, C _5-10 spiro ring, the alkyl group, cycloalkyl group, unsaturated cyclic hydrocarbon group, heterocycloalkyl group, aryl group, heteroaryl group, bridge The ring, paracyclic or spiro ring is optionally substituted with the following substituents: F, Cl, hydroxy, cyano, C _1-6 alkyl, hydroxy substituted C _1-6 alkyl, C _1-6 alkoxy, benzene ring, -C(O)OC _1-6 alkyl or C _3-6 cycloalkyl; in certain embodiments, R ¹ is selected from

,

,

, indolyl, pyrrolyl, pyridyl, oxidyl, phenyl, cyclohexyl, cyclopentyl, pyrazine, adamantyl, oxazine, morpholinyl,

,

,

,

,

or

; In certain embodiments, R ¹ is selected from

,

,

, indolyl, pyrrolyl, pyridyl, oxidyl, phenyl, cyclohexyl, cyclopentyl, pyrazine, adamantyl, oxazine, morpholinyl,

; In certain embodiments, R ¹ is selected from

,

,

,

; In certain embodiments, R ¹ is selected from

,

,

, adamantyl,

,

,

; In certain embodiments, R ¹ is selected from

,

,

,

,

,

; In certain embodiments, R ¹ is selected from

,

,

,

; In certain embodiments, R ¹ is selected from

,

,

; In certain embodiments, R ¹ is selected from

; In certain embodiments, R ¹ is selected from

; In certain embodiments, R ¹ is selected from

; R ² and R ³ are each independently selected from H, F, Cl, cyano, hydroxyl, C _1-6 alkoxy or C _1-6 alkyl; In certain embodiments, R ² and R ³ are each independently is selected from H, F, Cl, cyano, hydroxyl or C _1-6 alkyl; in certain embodiments, R ² , R ³ are each independently selected from H, F, Cl, methyl, ethyl, isopropyl In certain embodiments, R ² , R ³ are each independently selected from H, F or methyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H, F, Cl, amino, cyano, hydroxyl, C _1-6 alkyl or C _1-6 haloalkyl; in certain embodiments, R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H, F, cyano , hydroxy or C _1-6 alkyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H or C _1-6 alkyl; in certain embodiments, R ⁴ , R ⁵ , R ⁶ , R ⁷ is each independently selected from H, methyl, ethyl, and isopropyl; R ⁸ and R ⁹ are each independently selected from H, F, Cl, cyano, hydroxyl, C _1-6 alkoxy or C _1-6 Alkyl; R ⁸ , R ⁹ are each independently selected from H, F, Cl, C _1-6 alkyl, and the alkyl is optionally substituted by the following substituents: cyano, F or Cl, or R ⁸ and R ⁹ together form C _2-6 alkenyl or C _3-6 cycloalkyl; R ¹⁰ is selected from H or C _1-6 alkyl; R ¹¹ is selected from aryl or heteroaryl, said aryl or heteroaryl The group is optionally substituted with the following substituents: F, Cl, C _1-6 alkyl or C _1-6 alkoxy; in certain embodiments R ¹¹ is selected from

or

; In certain embodiments, R ¹¹ is selected from

; Condition is: when R ¹¹ is selected from aryl and substituted aryl, R ¹ is selected from

,

,

or

.

、

、

、吲哚基、吡咯基、吡啶基、呱啶基、苯基、環己基、環戊基、吡嗪、金剛烷基、呱嗪、嗎啉基、

、

、

、

、

或者、

。 The tetrahydroindazole compounds of the present invention and their stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs, wherein: R ¹ is selected from

,

,

, indolyl, pyrrolyl, pyridyl, oxidyl, phenyl, cyclohexyl, cyclopentyl, pyrazine, adamantyl, oxazine, morpholinyl,

,

,

,

,

or,

.

、

、

、

、

、

。 The tetrahydroindazole compounds of the present invention and their stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs, wherein: R ¹ is selected from

,

,

,

,

,

.

、

、

、吲哚基、吡咯基、吡啶基、呱啶基、苯基、環己基、環戊基、吡嗪、金剛烷基、呱嗪、嗎啉基、

、

、

、

、

、

或者

。 The tetrahydroindazole compounds of the present invention and their stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs, wherein: R ¹ is selected from

,

,

, indolyl, pyrrolyl, pyridyl, oxidyl, phenyl, cyclohexyl, cyclopentyl, pyrazine, adamantyl, oxazine, morpholinyl,

,

,

,

,

,

or

.

、

、

、吲哚基、吡咯基、吡啶基、呱啶基、 苯基、環己基、環戊基、吡嗪、金剛烷基、呱嗪、嗎啉基、

、

、

、

、

或者

。 The tetrahydroindazole compounds of the present invention and their stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs, wherein: R ¹ is selected from

,

,

, indolyl, pyrrolyl, pyridyl, oxidyl, phenyl, cyclohexyl, cyclopentyl, pyrazine, adamantyl, oxazine, morpholinyl,

,

,

,

,

or

.

或者

；條件是：當R¹¹選自

時，R¹選自

、

、

；在某些實施方案中，R¹¹選自

。 The tetrahydroindazole compounds of the present invention and their stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs, wherein: R ¹¹ is selected from

or

; the condition is: when R ¹¹ is selected from

, R ¹ is selected from

,

,

; In certain embodiments, R ¹¹ is selected from

.

或者

；條件是：當R¹¹選自

時，R¹選自

、

、

、

；在某些實施方案中，R¹¹選自

。 The tetrahydroindazole compounds of the present invention and their stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs, wherein: R ¹¹ is selected from

or

; the condition is: when R ¹¹ is selected from

, R ¹ is selected from

,

,

,

; In certain embodiments, R ¹¹ is selected from

.

或者

；條件是：當R¹¹選自

時，R¹選自

、

、

。 The tetrahydroindazole compounds of the present invention and their stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs, wherein: R ¹ is selected from C _1-6 alkyl Or C _3-6 cycloalkyl, the alkyl or cycloalkyl is optionally substituted by the following substituents: F, Cl, C _1-6 alkyl or hydroxyl; R ² and R ³ are each independently selected from H, F, Cl or C _1-6 alkyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H or C _1-6 alkyl; R ⁸ , R ⁹ are each independently selected from H, F, Cl, C _1-6 alkyl, said alkyl is optionally substituted by the following substituents: cyano, F or Cl, or R ⁸ and R ⁹ together form C _2-6 alkenyl or C _3-6 cycloalkyl; R ¹⁰ is selected from H; R ¹¹ is selected from

or

; the condition is: when R ¹¹ is selected from

, R ¹ is selected from

,

,

.

、

、

或者

；R²、R³各自獨立選自H、F或者甲基；R⁴、R⁵、R⁶、R⁷各自獨立選自H；R⁸、R⁹各自獨立選自H、F、異丙基、

，或者R⁸與R⁹一起形成2-甲基丙烯基或者環丙基；R¹⁰選自H；R¹¹選自

。 The tetrahydroindazole compounds of the present invention and their stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs, wherein: R ¹ is selected from

,

,

or

R ² , R ³ are each independently selected from H, F or methyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H; R ⁸ , R ⁹ are each independently selected from H, F, isopropyl ,

, or R ⁸ and R ⁹ together form 2-methylpropenyl or cyclopropyl; R ¹⁰ is selected from H; R ¹¹ is selected from

.

、

、

或者

； R²、R³各自獨立選自H、F或者甲基；R⁴、R⁵、R⁶、R⁷各自獨立選自H；R⁸、R⁹各自獨立選自H、F、異丙基，或者R⁸與R⁹一起形成2-甲基丙烯基或者環丙基；R¹⁰選自H；R¹¹選自

。 The tetrahydroindazole compounds of the present invention and their stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs, wherein: R ¹ is selected from

,

,

or

; R ² , R ³ are each independently selected from H, F or methyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H; R ⁸ , R ⁹ are each independently selected from H, F, isopropyl , or R ⁸ and R ⁹ together form 2-methylpropenyl or cyclopropyl; R ¹⁰ is selected from H; R ¹¹ is selected from

.

、

或者

；R²、R³各自獨立選自H、F或者甲基；R⁴、R⁵、R⁶、R⁷各自獨立選自H；R⁸、R⁹各自獨立選自H、F、異丙基、

，或者R⁸與R⁹一起形成2-甲基丙烯基或者環丙基；R¹⁰選自H；R¹¹選自

。 The tetrahydroindazole compounds of the present invention and their stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs, wherein: R ¹ is selected from

,

or

R ² , R ³ are each independently selected from H, F or methyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H; R ⁸ , R ⁹ are each independently selected from H, F, isopropyl ,

, or R ⁸ and R ⁹ together form 2-methylpropenyl or cyclopropyl; R ¹⁰ is selected from H; R ¹¹ is selected from

.

、

或者

；R²、R³各自獨立選自H、F或者甲基；R⁴、R⁵、R⁶、R⁷各自獨立選自H；R⁸、R⁹各自獨立選自H、F、異丙基； R¹⁰選自H；R¹¹選自

。 The tetrahydroindazole compounds of the present invention and their stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs, wherein: R ¹ is selected from

,

or

R ² , R ³ are each independently selected from H, F or methyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H; R ⁸ , R ⁹ are each independently selected from H, F, isopropyl ; R ¹⁰ is selected from H; R ¹¹ is selected from

.

The tetrahydroindazole compound of the present invention and its stereoisomer, pharmaceutically acceptable salt, solvate, hydrate, N-oxide or prodrug, wherein the compound structure is as follows:

The tetrahydroindazole compound of the present invention and its stereoisomer, pharmaceutically acceptable salt, solvate, hydrate, N-oxide or prodrug, wherein the compound structure is as follows:

The tetrahydroindazole compound of the present invention and its stereoisomer, pharmaceutically acceptable salt, solvate, hydrate, N-oxide or prodrug, wherein the compound structure is as follows:

The following compounds are preferred:

The present invention also relates to a pharmaceutical composition comprising: an effective dose of any of the compounds of the present invention and their stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or A prodrug, or further comprising one or more other therapeutic agents and a pharmaceutically acceptable carrier or excipient.

The present invention also relates to the tetrahydroindazole compound represented by the general formula (I) and its stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs or the compounds containing the present invention. Use of a composition in the manufacture of a medicament for the treatment of diseases mediated by CB2 receptors.

The present invention relates to tetrahydroindazole compounds represented by the general formula (I) and stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs thereof, or combinations containing the compounds of the present invention Use of a substance in the preparation of a medicament for the treatment of pain.

In some specific embodiments of the invention, the pain may be selected from the group consisting of: bone pain, arthralgia, muscle pain, dental pain, migraine, headache, inflammatory pain, neuropathic pain, Crohn's disease-related abdominal pain, adverse effects due to therapeutic agents Pain due to action and pain associated with a disease selected from the group consisting of osteoarthritis, cancer, multiple sclerosis, allergic response, nephritic syndrome, scleroderma, thyroiditis, diabetic neuropathy, fibromyalgia , HIV-related neuropathy, sciatica, and autoimmune diseases.

The present invention also relates to the tetrahydroindazole compound represented by the general formula (I) and its stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs or the compounds containing the present invention. Use of the composition in the preparation of a medicament for the treatment of neuropathic pain.

The present invention also relates to the tetrahydroindazole compound represented by the general formula (I) and its stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs or the compounds containing the present invention. Use of the composition in preparing a medicament for treating abdominal pain associated with Crohn's disease.

The present invention also relates to the tetrahydroindazole compound represented by the general formula (I) and its stereoisomer, The pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs or compositions containing the compounds of the present invention are useful in methods of treating diseases mediated by CB2 receptors.

The present invention also relates to the tetrahydroindazole compound represented by the general formula (I) and its stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs or the compounds containing the present invention. The composition is used in a method of treating pain.

In some embodiments of the invention, the pain may be selected from the group consisting of: bone pain, arthralgia, muscle pain, dental pain, migraine, headache, inflammatory pain, neuropathic pain, Crohn's disease-related abdominal pain due to adverse effects of therapeutic agents and pain associated with a disease selected from the group consisting of osteoarthritis, cancer, multiple sclerosis, allergic response, nephritic syndrome, scleroderma, thyroiditis, diabetic neuropathy, fibromyalgia, HIV-related neuropathy, sciatica, and autoimmune diseases.

The present invention also relates to the tetrahydroindazole compound represented by the general formula (I) and its stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs or the compounds containing the present invention. Compositions A method of treating neuropathic pain.

The present invention also relates to the tetrahydroindazole compound represented by the general formula (I) and its stereoisomers, pharmaceutically acceptable salts, solvates, hydrates, N-oxides or prodrugs or the compounds containing the present invention. Compositions for methods of treating abdominal pain associated with Crohn's disease.

Unless stated to the contrary, terms used in the specification and claims have the following meanings.

The carbon, hydrogen, oxygen, sulfur, nitrogen or halogen involved in the groups and compounds of the present invention all include their isotopes, and carbon, hydrogen, oxygen, sulfur, The nitrogen or halogen is optionally further replaced by one or more of their corresponding isotopes, wherein isotopes of carbon include12C, ^{13C and14C} , and isotopes of hydrogen include ^protium (H), deuterium (D, also known as deuterium) , tritium (T, also known as super heavy hydrogen), the isotopes of oxygen include ¹⁶ O, ¹⁷ O and ¹⁸ O, the isotopes of sulfur include ³² S, ³³ S, ³⁴ S and ³⁶ S, and the isotopes of nitrogen include ¹⁴ N and ¹⁵ N , the isotope of fluorine is ¹⁹ F, the isotope of chlorine includes ³⁵ Cl and ³⁷ Cl, and the isotope of bromine includes ⁷⁹ Br and ⁸¹ Br.

"Alkyl" refers to straight and branched monovalent saturated hydrocarbon groups, the main chain comprising 1 to 10 carbons atoms, preferably 1 to 8 carbon atoms, more preferably 1 to 6 carbon atoms, more preferably 1 to 4 carbon atoms, straight and branched chain groups, most preferably 1 to 2 carbon atoms , examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-pentyl, 3- pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, n-hexyl, n-heptyl, n-octyl, n-nonyl and n-decyl, etc.; the alkyl group can be further Any substituent is substituted.

"Alkylene" refers to linear and branched divalent saturated hydrocarbon groups, including -(CH ₂ ) _v - (v is an integer from 1 to 10). Examples of alkylene include but are not limited to methylene, methylene Ethyl, propylene and butylene, etc.; the alkylene group may be optionally further substituted by any substituent. When the number of substituents in the alkylene group is 2 or more, the substituents can be fused together to form a cyclic structure.

"Alkoxy" refers to a monovalent group of O-alkyl (-O-alkyl), wherein alkyl is as defined herein, and examples of alkoxy include, but are not limited to, methoxy, ethoxy, 1-propoxy, 2-propoxy, 1-butoxy, 2-methyl-1-propoxy, 2-butoxy, 2-methyl-2-propoxy, 1-pentyloxy yl, 2-pentyloxy, 3-pentyloxy, 2-methyl-2-butoxy, 3-methyl-2-butoxy, 3-methyl-1-butoxy and 2-methyl base-1-butoxy, etc.

"Alkenyl" refers to linear and branched monovalent unsaturated hydrocarbon groups, which have at least 1, usually 1, 2 or 3 carbon-carbon double bonds, and the main chain includes 2 to 10 carbon atoms, more preferably 2 to 6 carbon atoms, more preferably 2 to 4 carbon atoms in the main chain, examples of alkenyl groups include but are not limited to vinyl, allyl, 1-propenyl, 2-propenyl, 1-butene base, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2 -Methyl-1-butenyl, 2-methyl-3-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl , 1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 1-octenyl Alkenyl, 3-octenyl, 1-nonenyl, 3-nonenyl, 1-decenyl, 4-decenyl, 1,3-butadiene, 1,3-pentadiene, 1 ,4-pentadiene and 1,4-hexadiene, etc.; the alkenyl group can be optionally further substituted by any group.

"Alkynyl" refers to linear and branched monovalent unsaturated hydrocarbon groups, which have at least 1, usually 1, 2 or 3 carbon-carbon triple bonds, and the main chain includes 2 to 10 carbon atoms, more preferably 2 to 6 carbon atoms, more preferably 2 to 4 carbon atoms in the main chain, examples of alkynyl groups include but are not limited to ethynyl, 1-propane alkynyl, 2-propynyl, butynyl, 2-butynyl, 3-butynyl, 1-methyl-2-propynyl, 4-pentynyl, 3-pentynyl, 1- Methyl-2-butynyl, 2-hexynyl, 3-hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octynyl, 3-nonynyl and 4-decynyl, etc.; the alkynyl group can be optionally further substituted by any substituent.

"Cycloalkyl" means a monovalent saturated carbocyclic hydrocarbon group, monocyclic, usually having 3 to 10 carbon atoms, non-limiting examples including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl Base et al. The cycloalkyl group may be optionally further substituted with any substituent.

"Unsaturated cyclic hydrocarbon group" refers to a carbocyclic hydrocarbon group containing an unsaturated bond (such as a double bond) without aromaticity, non-limiting examples include cyclopropenyl, cyclobutenyl, 1,4-cyclohexanedi alkene, 1,3-cyclopentadiene, etc. The cycloalkyl group may be optionally further substituted with any substituent.

"Heterocyclyl" refers to a saturated or unsaturated cyclic hydrocarbon group containing at least one heteroatom, a single ring, the heteroatoms are N, O, S, P and their oxidized forms, non-limiting examples include aza Cyclopropyl, oxetanyl, thietanyl, azetidine, oxetanyl, thietanyl, azetidine (aka azetidine), pyrrolyl, Pyrrolinyl, 3-pyrrolinyl, 1-pyrrolinyl, pyrrolidinyl, pyrazolidinyl, 2-pyrazolinyl, imidazolyl, pyrazolyl, tetrahydrofuranyl, tetrahydrothienyl, 1,2 , 4-triazolyl, etc. Said heterocyclic group may be optionally further substituted with any substituent.

和

。螺環可以被任意取代基進一步取代。 "Spirocycle" refers to a 5- to 20-membered polycyclic group of substituted or unsubstituted monocyclic rings sharing one atom (such as a carbon atom, referred to as a spiro atom), which may contain 0 to 5 double bonds, and may Contains 0 to 5 heteroatoms selected from N, O or S(=O)n. Preferably it is 6 to 14 yuan, more preferably 6 to 12 yuan, more preferably 6 to 10 yuan, non-limiting examples thereof include

and

. The spiro ring can be further substituted with any substituent.

和

。並環可以進一步被任意取代基取代。 "Paracyclic" refers to a polycyclic group in which each ring in the system shares an adjacent pair of atoms (such as carbon atoms) with other rings in the system, one or more of which may contain zero or more double bonds , and may be substituted or unsubstituted, and each ring in the ring system may contain 0 to 5 heteroatoms selected from N, S(=O) _n , or O. Preferably it is 5 to 20 yuan, more preferably 5 to 14 yuan, more preferably 5 to 12 yuan, and still more preferably 5 to 10 yuan. Non-limiting examples include

and

. The parallel ring may be further substituted with any substituent.

和金剛烷。橋環可以進一步被任意取代基取代。 "Bridged ring" refers to a polycyclic group in which any two rings share two atoms (such as carbon atoms) that are not directly connected, may contain 0 or more double bonds, and may be substituted or unsubstituted, and Any ring in the ring system may contain 0 to 5 heteroatoms or groups selected from N, S(=O)n or O (wherein n is 1, 1, 2). The ring atoms contain 5 to 20 atoms, preferably 5 to 14 atoms, more preferably 5 to 12 atoms, still more preferably 5 to 10 atoms. Non-limiting examples include

and adamantane. The bridged ring may be further substituted with any substituent.

、

、

、

、

、

和

。芳基可以進一步被任意取代基取代。 "Aryl" refers to a substituted or unsubstituted 6- to 14-membered cyclic aromatic group, including monocyclic aromatic groups and fused-ring aromatic groups. A 6- to 14-membered aromatic ring is preferable, and a 6- to 10-membered aromatic ring is further preferable, and non-limiting examples thereof include phenyl, naphthyl, anthracenyl, and phenanthryl, and the like. The aryl ring can be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring linked to the parent structure is an aryl ring, non-limiting examples include:

,

,

,

,

,

and

. The aryl group may be further substituted with any substituent.

、

、

、

、

、

、

和

。雜芳基可以進一步被任意取代基取代。 "Heteroaryl" refers to a substituted or unsubstituted 5- to 14-membered aromatic ring, and contains 1 to 5 heteroatoms or groups selected from N, O or S(=O) _n , preferably a 5- to 10-membered heteroatom Aromatic ring, more preferably 5 to 6 members. Non-limiting examples of heteroaryl groups include, but are not limited to, pyridyl, furyl, thienyl, pyridyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, Ciridinyl, morpholine, thiomorpholine, 1,3-dithiane, benzimidazole, benzopyridine, pyrrolopyridine, etc. The heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring linked to the parent structure is a heteroaryl ring, non-limiting examples include

,

,

,

,

,

,

and

. Heteroaryl groups may be further substituted with optional substituents.

"Pharmaceutically acceptable salt" means one that retains the biological availability and properties of a free acid or free base that has been treated with a non-toxic inorganic or organic base, or the free base that has been treated with a non-toxic Those salts obtained by the reaction of inorganic or organic acids.

"Carrier" refers to a carrier or diluent that is not appreciably irritating to the organism and that does not abrogate the biological activity and properties of the administered compound.

"Excipient" refers to an inert substance added to a pharmaceutical composition to further depend upon the administration of the compound. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and different types of starch, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulation agents, lubricants, binders, disintegrants, etc.

"Prodrug" refers to a compound that can be converted under physiological conditions or by solvolysis to a biologically active compound of the present invention. The prodrugs of the present invention are prepared by modifying the phenolic group in the compound, which modification can be removed by conventional procedures or in vivo to yield the parent compound. When the prodrugs of the present invention are administered to a mammalian subject, the prodrugs are cleaved to form free hydroxyl groups, respectively. Examples of prodrugs include, but are not limited to, the phenolic hydroxyl and phosphate sodium derivatives of the compounds of the present invention.

"Effective dose" refers to a compound that elicits a physiological or medical response in a tissue, system, or subject The amount sought, including that amount of the compound, when administered to a subject, is sufficient to prevent or reduce to some extent one or more symptoms of the disorder or disorder being treated.

"Solvates" refer to compounds of the present invention or salts thereof, which also include stoichiometric or non-stoichiometric amounts of solvent bound by intermolecular non-covalent forces. When the solvent is water, it is a hydrate.

"Optional" or "optionally" means that the subsequently described event or circumstance can, but need not, occur, and that the description includes instances where the event or circumstance does or does not occur. For example, "alkyl optionally substituted by F" means that the alkyl group may but not necessarily be substituted by F, and the description includes the case where the alkyl group is substituted by F and the case where the alkyl group is not substituted by F.

"Optionally substituted by R" means that it can be substituted by R or not. When substituted, the number of R is not limited, and the principle of chemical bond is satisfied, preferably 1-5, 1-3, 1-2 , 1; when substituted by two or more Rs, R can be independently and freely selected, and can be the same or different, and are independent of each other.

The technical solutions of the present invention are described in detail below with reference to the embodiments, but the protection scope of the present invention includes but is not limited thereto.

The structures of the compounds were determined by nuclear magnetic resonance (NMR) and/or mass spectrometry (MS).

NMR shifts ([delta]) are given in units of 10-6 (ppm).

NMR was measured with a (Bruker ADVANCE III 400) nuclear magnetic instrument, and the solvent was deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD), and the internal standard was Tetramethylsilane (TMS), ¹ HNMR information is listed in the following format: chemical shift (multiplet (s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet), number of protons).

For MS measurement (Agilent 6120B (ESI)).

The HPLC determination was performed using an Agilent 1260DAD high pressure liquid chromatograph (Zorba x SB- C18 100 x 4.6mm).

The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, the size of the silica gel plate used for thin layer chromatography (TLC) is 0.15mm~0.20mm, and the specification used for TLC separation and purification products is 0.4mm ~0.5mm.

Column chromatography generally uses Yantai Huanghai silica gel 200~300 mesh silica gel as the carrier.

There is no special description in the examples, and the solution refers to an aqueous solution.

There is no special description in the examples, and the temperature of the reaction is room temperature, which is 20°C to 30°C.

Short description:

HATU: 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate; DMF: N,N-dimethylformamide; TEA: tris ethylamine; LHMDS: lithium bis(trimethylsilyl)amide; DCM: dichloromethane; m-CPBA: m-chloroperoxybenzoic acid; DIPEA: N,N-diisopropylethylamine; PE: petroleum ether; EA: ethyl acetate; NIS: N-iodosuccinimide.

Intermediate Int-1

2-Amino-4,4,4-trifluoro-3,3-dimethylbutan-1-ol (Intermediate Int-1 ) 2-amino-4,4,4-trifluoro-3,3-dimethylbutan -1-ol

The first step: 3,3,3-trifluoro-N-methoxy-N,2,2-trimethylpropionamide (Int-1-b)

3,3,3-trifluoro-N-methoxy-N,2,2-trimethylpropanamide

Under nitrogen protection, at room temperature, to 3,3,3-trifluoro-2,2-dimethylpropionic acid (25g, 0.16mol), were added DMF (50mL), TEA (32mg, 0.25mmol), HATU successively (77 mg, 0.2 mmol), N-methyl-N-methoxyamine hydrochloride (17.2 g, 0.176 mol), and stirred at room temperature overnight. Saturated aqueous sodium bicarbonate solution (200 mL) was added to the reaction to quench the reaction, extracted with 300 mL of ethyl acetate, left to stand for layers, the aqueous phase was extracted three times with dichloromethane (200 mL×3), and the combined organic phases were extracted with anhydrous sulfuric acid After drying over sodium, the residue was obtained after concentration under reduced pressure, that is, the crude product. The crude product was subjected to column chromatography (eluent PE:EA=10:1) to obtain Int-1-b (22 g, yield 69%), pale yellow oily .

LC-MS (ESI): m/z=200.2 [M+H] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ) δ 3.70 (s, 3H), 3.21 (s, 3H), 1.5 (s, 6H).

The second step: 3,3,3-trifluoro-2,2-dimethylpropanal ( Int-1-c )

3,3,3-trifluoro-2,2-dimethylpropanal

Under nitrogen protection, at room temperature, THF (50 mL) was added to a 250 mL single-neck flask, LiAlH (3.43 g, 90 mmol) was added at 0° C., and compound Int-1-b ₍ 15 g, 75.3 mmol) was slowly added dropwise, and stirred at room temperature. 2h. At 0°C, water (3.5 mL), NaOH (15%, 3.5 mL), and water (3.5 mL) were sequentially added to the reaction to quench the reaction, and the reaction was stirred for 30 min. Then 200 mL of ether was added and vigorously stirred for 30 min, and filtered through celite. , and concentrated under reduced pressure to obtain crude Int-1-c (14 g), a colorless oily compound, which was used in the next reaction without purification.

LC-MS (ESI): m/z=141.2 [M+H] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ) δ 9.69 (s, 1H), 1.30 (s, 6H).

The third step: 2-amino-4,4,4-trifluoro-3,3-dimethylbutyronitrile ( Int-1-d )

2-amino-4,4,4-trifluoro-3,3-dimethylbutanenitrile

Under nitrogen protection, at room temperature, sequentially add Int-1-c (4.5g), NH ₃ /MeOH (10 mL, 70 mmol), NH ₄ Cl (1.62 g, 30 mmol) to the single-necked flask, stir at room temperature for 30 min, and then KCN (1.95 g, 30 mmol) was added and the reaction continued to stir overnight. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was dissolved in 10 mL of ether, and 2N HCl was slowly added at 0 °C. 1,4-dioxane was dissolved and stirred to precipitate a white solid Int-1-d (1.65 g, yield 34.78%).

LC-MS (ESI): m/z=167.1 [M+H] ⁺ .

The fourth step: 2-amino-4,4,4-trifluoro-3,3-dimethylbutyric acid ( Int-1-e )

2-amino-4,4,4-trifluoro-3,3-dimethylbutanoic acid

Under nitrogen protection, at room temperature, Int-1-d (2.05 g, 10.14 mmol) and 6N HCl (20 mL) were successively added to the single-necked flask, and the mixture was stirred under reflux for 36 h. Cool to room temperature, add aqueous sodium hydroxide solution to the reaction to adjust pH to 8, the aqueous phase is washed with dichloromethane (20 mL) to recover 0.4 g of raw materials, the aqueous phase is adjusted to pH 7 again, and concentrated to solid under reduced pressure to obtain a crude white product Solid Int-1-e (1.3 g).

LC-MS (ESI): m/z=186.2 [M+H] ⁺ .

The fifth step: 2-amino-4,4,4-trifluoro-3,3-dimethylbutan-1-ol ( Int-1 )

2-amino-4,4,4-trifluoro-3,3-dimethylbutan-1-ol

Under nitrogen protection, THF (20 mL) was added to a 50 mL single-neck flask at room temperature, LiAlH ₄ (270 mg, 8.43 mmol) was added at 0° C., and compound Int-1-e (1.3 g, 7 mmol) was slowly added dropwise, and stirred for 3 h under reflux. The reaction is complete. At 0 °C, water (0.27 mL), NaOH (15%, 0.27 mL), and water (0.27 mL) were successively added to the reaction to quench the reaction, stirred for 30 min, and then THF (50 mL) was added to vigorously stir for 30 min, and dried over NaSO ₄ , filtered through celite, concentrated under reduced pressure to obtain Int-1 (0.82 g, 68.23%), a pale yellow oily compound, which was directly used in the next reaction without purification.

LC-MS (ESI): m/z=172.2 [M+H] ⁺ .

Example 1

3-((4R)-3-((s)-1-Hydroxy-3,3-dimethylbutan-2-ylaminocarboxy)-7-isopropyl-4-methyl-4, 5,6,7-Tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide (Compound 1)

3-((4R)-3-(((S)-1-hydroxy-3,3-dimethylbutan-2-yl)carbamoyl)-7-isopropyl-4-methyl-4,5,6,7-tetrahydro- 1H-indazol-1-yl)pyrazine 1-oxide

The first step: (2S,5R)-2-isopropyl-5-methylcyclohexanone

(2S,5R)-2-isopropyl-5-methylcyclohexanone ( 1b )

Dichloromethane (100 mL) was added to known compound 1a (5.0 g, 32.0 mmol) at room temperature, Dess Martin oxidizing agent (13.6 g, 32 mmol) was added in batches under water bath cooling, and the mixture was stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate solution (100 mL) was added to the reaction to quench the reaction, and the reaction was stirred at room temperature for 30 minutes. Filter through a pad of diatomaceous earth, leave the mother liquor to stand for stratification, the aqueous phase is back-extracted with dichloromethane (100 mL), the combined organic phases are dried over anhydrous sodium sulfate, and the residue is concentrated under reduced pressure with silica gel column chromatography (eluent). PE:EA=20:1-5:1) was isolated and purified to obtain 1b (3.4g, 69%).

LC-MS (ESI): m/z=155.2 [M+H] ⁺ .

The second step: (4R)-7-isopropyl-4-methyl-3a,4,5,6-tetrahydrobenzofuran-2,3-dione ( 1c )

(4R)-7-isopropyl-4-methyl-3a,4,5,6-tetrahydrobenzofuran-2,3-dione

At room temperature, MeOH (10 mL), dimethyl oxalate (0.92 g, 7.78 mmol), and 30% sodium methoxide solution (5 ml, 89.2 mmol) were sequentially added to compound 1b (1.0 g, 6.48 mmol). The reaction was stirred at 50°C for 18 hours. The pH of the reaction solution was adjusted to 5-6 with 1M hydrochloric acid, diluted with water (100 mL), extracted with ethyl acetate (100 mL), left to stand for layers, the organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was washed with silica gel 1c (0.84g, 62%) was isolated and purified by column chromatography (eluent PE:EA=10:1-5:1).

LC-MS (ESI): m/z=209.2 [M+H] ⁺ .

The third step: 2-((6R)-3-isopropyl-6-methyl-2-oxocyclohexyl)-2-oxoacetic acid ( 1d )

2-((6R)-3-isopropyl-6-methyl-2-oxocyclohexyl)-2-oxoacetic acid

At room temperature, tetrahydrofuran (5 mL), water (5 mL), NaOH (2.0 g, 50 mmol) were sequentially added to compound 1c (1.0 g, 4.8 mmol), stirred for 2 hours, adjusted to pH 5-6 with 1 M HCl, acetic acid Extracted with ethyl ester, the ethyl acetate phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 1d (0.8 g, 70%) as the residue.

LC-MS (ESI): m/z=227.2 [M+H] ⁺ .

The fourth step: (4R)-7-isopropyl-4-methyl-1-(pyrazin-2-yl)-4,5,6,7-tetrahydro-1H-indazole-3-carboxylic acid ( 1e )

(4R)-7-isopropyl-4-methyl-1-(pyrazin-2-yl)-4,5,6,7-tetrahydro-1H-indazole-3-carboxylic acid

At room temperature, MeOH (50 mL), 2-hydrazinopyrazine (0.5 g, 5.0 mmol) were sequentially added to compound 1d (0.8 g, 4.0 mmol), and the pH was adjusted to 1-2 with 6M hydrochloric acid. Stir at 50 °C After 2 hours, diluted with water, extracted with ethyl acetate, the ethyl acetate phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (eluent PE:EA=5: 1-pure EA) was isolated and purified to give 1e (0.2 g, 16.7%).

LC-MS (ESI): m/z=301.2 [M+H] ⁺ .

The fifth step: (4R)-N-((S)-1-hydroxy-3,3-dimethylbutan-2-yl)-7-isopropyl-4-methyl-1-(pyrazine -2-yl)-4,5,6,7-tetrahydro-1H-indazole-3-carboxamide ( 1f )

(4R)-N-((S)-1-hydroxy-3,3-dimethylbutan-2-yl)-7-isopropyl-4-methyl-1-(pyrazin-2-yl)-4,5,6, 7-tetrahydro-1H-indazole-3-carboxamide

At room temperature, to compound 1e (0.2 g, 0.66 mmol) were sequentially added DMF (2 mL), HATU (0.28 g, 0.73 mmol), (S)-2-amino-3,3-dimethylbutanol ( 0.085 g, 0.72 mmol), triethylamine (0.2 g, 1.98 mmol). The mixture was stirred at room temperature for 2 hours, diluted with water, and extracted with ethyl acetate. The ethyl acetate phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (eluent PE:EA). =5:1-2:1) separation and purification to obtain 1f (0.2g, 75.5%).

LC-MS (ESI): m/z=400.3 [M+H] ⁺ .

The sixth step: 3-((4R)-3-(((S)-1-hydroxy-3,3-dimethylbutan-2-yl)amine carboxamide)-7-isopropyl-4- Methyl-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide ( Compound 1 )

3-((4R)-3-(((S)-1-hydroxy-3,3-dimethylbutan-2-yl)carbamoyl)-7-isopropyl-4-methyl-4,5,6,7-tetrahydro- 1H-indazol-1-yl)pyrazine 1-oxide

At room temperature, DCM (1 mL) and m-CPBA (0.065 g, 0.376 mmol) were sequentially added to compound 1f (0.05 g, 0.125 mmol), and the mixture was stirred at room temperature for 18 hours. The reaction was quenched with aqueous sodium sulfite solution, diluted with water, and acetic acid. Extraction with ethyl ester, the ethyl acetate phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (PE:EA=5:1-1:1) to obtain compound 1 (0.01 g, 19.2%).

LC-MS (ESI): m/z=416.3 [M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD) δ 8.99-8.98 (m, 1H), 8.43-8.42 (d, 1H), 8.22-8.21 (m, 1H), 3.97-3.94 (m, 1H), 3.86-3.82 (m,1H),3.69-3.66(m,2H),3.28-3.25(m,1H),2.14-2.06(m,1H),1.86-1.76(m,3H),1.69-1.66(m,1H) , 1.29-1.27(d, 3H), 1.02(s, 9H), 0.94-0.92(d, 3H), 0.70-0.68(d, 3H).

Example 2

(4R)-1-(2,4-Difluorophenyl)-N-((S)-1-hydroxy-3,3-dimethylbutan-2-yl)-7-isopropyl-4 - Methyl-4,5,6,7-tetrahydro-1H-indazole-3-carboxamide (compound 2 ).

(4R)-1-(2,4-difluorophenyl)-N-((S)-1-hydroxy-3,3-dimethylbutan-2-yl)-7-isopropyl-4-methyl-4,5,6, 7-tetrahydro-1H-indazole-3-carboxamide.

The first step: (4R)-1-(2,4-difluorophenyl)-7-isopropyl-4-methyl-4,5,6,7-tetrahydro-1H-indazole-3- Formic acid ( 2a ).

(4R)-1-(2,4-difluorophenyl)-7-isopropyl-4-methyl-4,5,6,7-tetrahydro-1H-indazole-3-carboxylic acid.

At room temperature, methanol (6 mL) and 2,4-difluorophenylhydrazine (0.22 g, 1.6 mmol) were sequentially added to compound 1d (0.3 g, 1.3 mmol), and the pH value was adjusted to 3-4 with hydrochloric acid (6 M), Heat to 50°C and stir the reaction for 2 hours. The reaction solution was poured into (30 mL) water, extracted with ethyl acetate (20 mL×2), the organic phase was concentrated, and separated and purified by column chromatography (eluent PE/EA=10/1-5/1) to obtain compound 2a (0.2 g, 46% yield).

LC-MS (ESI): m/z=335.2 [M+H] ⁺ .

The second step: (4R)-1-(2,4-difluorophenyl)-N-((S)-1-hydroxy-3,3-dimethylbutan-2-yl)-7-iso Propyl-4-methyl-4,5,6,7-tetrahydro-1H-indazole-3-carboxamide ( compound 2 ).

(4R)-1-(2,4-difluorophenyl)-N-((S)-1-hydroxy-3,3-dimethylbutan-2-yl)-7-isopropyl-4-methyl-4,5,6, 7-tetrahydro-1H-indazole-3-carboxamide.

At room temperature, to compound 2a (0.1 g, 0.3 mmol) were sequentially added DMF (5 mL), S-tert-leucinol (39 mg, 0.33 mmol), HATU (125 mg, 0.33 mmol), DIPEA (120 mg, 0.9 mmol) . The reaction was stirred for 1 hour. The reaction solution was poured into (50 mL) water, extracted with ethyl acetate (20 mL×2), the organic phase was concentrated, and separated and purified by silica gel column chromatography (eluent PE/EA=5/1-1/1) to obtain compound 2 (25 mg, 19% yield).

LC-MS (ESI): m/z=434.2 [M+H] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.45-7.39 (m, 1H), 7.03-6.97 (m, 2H), 3.99-3.89 (m, 2H), 3.62-3.57 (m, 1H), 3.41-3.99 ( m, 1H), 2.88-2.83(m, 1H), 1.80-1.68(m, 4H), 1.64-1.56(m, 1H), 1.31-1.29(d, 3H), 1.02(s, 9H), 0.81- 0.79(d, 3H), 0.58-0.56(d, 3H).

Example 3

3-((4R)-3-((1-Hydroxy-2-methylpropanol-2-yl)aminocarboxy)-7-isopropyl-4-methyl-4,5,6,7- Tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide (compound 3 ).

3-((4R)-3-((1-hydroxy-2-methylpropan-2-yl)carbamoyl)-7-isopropyl-4-methyl-4,5,6,7-tetrahydro-1H-indazol-1- yl)pyrazine 1-oxide.

The first step: 3-((4R)-3-carboxy-7-isopropyl-4-methyl-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine-1 - oxide ( 3a ).

3-((4R)-3-carboxy-7-isopropyl-4-methyl-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide.

At room temperature, methanol (8 mL) and pyrazine-2-hydrazine oxide (0.27 g, 2.1 mmol) were sequentially added to compound 1d (0.4 g, 1.8 mmol), and the pH value was adjusted to 3-4 with hydrochloric acid (6 M), Heat to 50°C and stir the reaction for 2 hours. The reaction solution was poured into (30 mL) water, extracted with ethyl acetate (20 mL×2), the organic phase was concentrated, and separated and purified by column chromatography (eluent DCM/MeOH=80/1-30/1) to obtain compound 3a (0.12 g, 21% yield).

LC-MS (ESI): m/z=317.2 [M+H] ⁺ .

The second step: 3-((4R)-3-((1-hydroxy-2-methylpropanol-2-yl)aminocarbamide)-7-isopropyl-4-methyl-4,5, 6,7-Tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide ( compound 3 ).

3-((4R)-3-((1-hydroxy-2-methylpropan-2-yl)carbamoyl)-7-isopropyl-4-methyl-4,5,6,7-tetrahydro-1H-indazol-1- yl)pyrazine 1-oxide.

At room temperature, to compound 3a (120 mg, 0.38 mmol) were sequentially added DMF (5 mL), 2-amino-2-methyl-1-propanol (37 mg, 0.42 mmol), HATU (160 mg, 0.42 mmol), DIPEA (147 mg, 1.14 mmol). The reaction was stirred for 1 hour. The reaction solution was poured into (50 mL) water, extracted with ethyl acetate (20 mL×2), the organic phase was concentrated, and separated and purified by silica gel column chromatography (eluent PE/EA=5/1-1/1) to obtain compound 3 (30 mg, 20% yield).

LC-MS (ESI): m/z=388.2 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ )δ 8.78(s,1H), 8.28-8.27(m,1H), 8.02-8.01(m,1H), 6.98(s,1H), 3.75-3.70(m,2H) ,3.64-3.60(m,1H),3.37-3.31(m,2H),1.80-1.73(m,4H),1.42-1.41(m,6H),1.31-1.29(m,3H),0.92-0.90( m, 3H), 0.65-0.63 (m, 3H).

Example 4:

(4R)-N-(1-(hydroxymethyl)cyclopentyl)-7-isopropyl-4-methyl-1-(pyrazin-2-yl)-4,5,6,7-tetra Hydro-1H-indazole-3-carboxamide-oxide (compound 4)

(4R)-N-(1-(hydroxymethyl)cyclopentyl)-7-isopropyl-4-methyl-1-(pyrazin-2-yl)-4,5,6,7-tetrahydro-1H-indazole-3-carboxamide -oxide

The first step: (4R)-N-(1-(hydroxymethyl)cyclopentyl)-7-isopropyl-4-methyl-1-(pyrazin-2-yl)-4,5,6 ,7-Tetrahydro-1H-indazole-3-carboxamide-oxide ( Compound 4 )

(4R)-N-(1-(hydroxymethyl)cyclopentyl)-7-isopropyl-4-methyl-1-(pyrazin-2-yl)-4,5,6,7-tetrahydro-1H-indazole-3-carboxamide -oxide

At room temperature, to compound 3a (0.1 g, 0.316 mmol) were sequentially added DCM (2 mL), HATU (0.14 g, 0.35 mmol), (1-aminocyclopentyl) methanol (0.045 g, 0.38 mmol), three Ethylamine (0.1 g, 0.95 mmol). The mixture was stirred at room temperature for 2 hours, diluted with water, and extracted with ethyl acetate. The ethyl acetate phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (eluent PE/EA). =5/1-1/1) was isolated and purified to obtain compound 4 (0.06g, 50%).

LC-MS (ESI): m/z=414.3 [M+H] ⁺ .

¹ H NMR (400MHz, CD ₃ OD) δ 8.79-8.72 (m, 1H), 8.26-8.24 (m, 1H), 8.01-7.98 (m, 1H), 7.07-7.06 (m, 1H), 3.80-3.72 (m,2H),3.66-3.59(m,1H),3.39-3.29(m,1H),2.06-1.96(m,3H),1.92-1.64(m,9H),1.31-1.22(m,3H) ,0.92-0.83(m,3H),0.82-0.60(m,3H).

Example 5

(S)-3-(7,7-Difluoro-3-((1-hydroxy-3,3-dimethylbutan-2-yl)aminocarbamoyl)-4,5,6,7- Tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide (Compound 5)

(S)-3-(7,7-difluoro-3-((1-hydroxy-3,3-dimethylbutan-2-yl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1- yl)pyrazine 1-oxide

The first step: 2-ethoxycyclohexane-2-en-1-one ( 5b )

2-ethoxycyclohex-2-en-1-one

Compound 5a (10 g, 89.3 mmol) was dissolved in toluene (100 mL), ethanol (50 mL) and p-toluenesulfonic acid (23.1 g, 133.0 mmol) were sequentially added thereto at room temperature. The temperature was raised to reflux for 24 hours. The reaction solution was spin-dried, mixed with silica gel, and separated by column chromatography (eluent: petroleum ether/ethyl acetate=10:1~5:1) to obtain compound 5b (6.5 g, 52%).

LC-MS (ESI): m/z=141.2 [M+H] ⁺ .

The second step: 2-(3-ethoxy-2-oxocyclohex-3-en-1-yl)-2-oxoethyl acetate ( 5c )

ethyl 2-(3-ethoxy-2-oxocyclohex-3-en-1-yl)-2-oxoacetate

Compound 5b (3.0 g, 21.4 mmol) was dissolved in tetrahydrofuran (30 mL), the temperature was lowered to -78 degrees Celsius, LHMDS (1 M, 23.5 mL) was added to it, and diethyl oxalate (3.4 g) was added to it after half an hour. , 23.5mmol), warmed to room temperature and reacted overnight. The reaction was quenched with dilute hydrochloric acid, the aqueous phase was extracted with ethyl acetate (100 mL×2), dried over anhydrous sodium sulfate, and then added with silica gel for column chromatography (eluent: petroleum ether/ethyl acetate = 10:1~5:1 ) was isolated to give compound 5c (1.9 g, 37.0%).

LC-MS (ESI): m/z=241.2 [M+H] ⁺ .

The third step: (7-oxo-1-(pyrazin-2-yl)-4,5,6,7-tetrahydro-1H-indazole-3-carboxylic acid ethyl ester ( 5d )

ethyl 7-oxo-1-(pyrazin-2-yl)-4,5,6,7-tetrahydro-1H-indazole-3-carboxylate

Compound 5c (2.1 g, 8.8 mmol) was dissolved in a sulfuric acid ethanol solution (1:100 (v/v), 21 mL), to which was added pyrazine-2-hydrazine (1.45 g, 13.2 mmol) at room temperature, The temperature was raised and refluxed for half an hour. The reaction solution was spin-dried, and then silica gel mixed sample column chromatography (eluent: petroleum ether/ethyl acetate=5:1~3:1) was added for separation to obtain compound 5d (1.85g, 73.4%).

¹ H NMR (400MHz, Chloroform-d)δ 9.16(d,1H), 8.64(d,1H), 8.55(dd,1H), 4.45(q,2H), 2.95(t,2H), 2.37-2.20( m, 2H), 2.16-2.03 (m, 2H), 1.43 (t, 3H).

The fourth step: 1-(pyrazin-2-yl)-1,4,5,6-tetrahydrospiro[indazole-7,2'-[1,3]dithiolane]-3-carboxylic acid Ethyl ester ( 5e )

ethyl 1-(pyrazin-2-yl)-1,4,5,6-tetrahydrospiro[indazole-7,2'-[1,3]dithiolane]-3-carboxylate

Compound 5d (0.9 g, 3.15 mmol) was dissolved in anhydrous dichloromethane (10 mL), and boron trifluoride ether solution (2M, 10 mL), ethanedithiol (1.45 g, 13.2 mmol) were sequentially added thereto at room temperature ) and reacted overnight at room temperature. The reaction was quenched with saturated sodium bicarbonate solution, the aqueous phase was extracted with ethyl acetate (70mL×2), dried over anhydrous sodium sulfate, added with silica gel and mixed with the sample, column chromatography (eluent: petroleum ether/ethyl acetate=5:1~ 3:1) was isolated to obtain compound 5e (0.9 g, 78.7%).

LC-MS (ESI): m/z=363.3 [M+H] ⁺ .

Fifth step: 7,7-difluoro-1-(pyrazin-2-yl)-4,5,6,7-tetrahydro-1H-indazole-3-carboxylic acid ethyl ester ( 5f )

ethyl 7,7-difluoro-1-(pyrazin-2-yl)-4,5,6,7-tetrahydro-1H-indazole-3-carboxylate

NIS (0.9 g, 2.49 mmol) was dissolved in anhydrous dichloromethane (10 mL), the temperature was lowered to minus 78 degrees Celsius, a solution of pyridinium hydrofluoride was added thereto, and after ten minutes of reaction, compound 5e (0.9 g, 2.49 mmol) in dichloromethane (10 mL) and reacted at this temperature for twenty minutes. The reaction solution was spin-dried, added with silica gel to mix samples, and separated by column chromatography (eluent: petroleum ether/ethyl acetate=2:1~1:1) to obtain compound 5f (0.48g, 47%).

LC-MS (ESI): m/z=309.1 [M+H] ⁺ .

Step 6: 7,7-Difluoro-1-(pyrazin-2-yl)-4,5,6,7-tetrahydro-1H-indazole-3-carboxylic acid ( 5g )

7,7-difluoro-1-(pyrazin-2-yl)-4,5,6,7-tetrahydro-1H-indazole-3-carboxylic acid

Compound 5f (0.15 g, 0.54 mmol) was dissolved in a mixed solvent of tetrahydrofuran (10 mL) and methanol (10 mL), and thereto was added sodium hydroxide solution (2M, 10 mL) at room temperature. The reaction was carried out at room temperature for two hours. The pH of the reaction solution was adjusted to 6, the aqueous phase was extracted with dichloromethane (30 mL×2), and the compound 5g was obtained by spin drying, which was used for later use without purification.

LC-MS (ESI): m/z=281.2 [M+H] ⁺ .

The seventh step: (S)-7,7-difluoro-N-(1-hydroxy-3,3-dimethylbutan-2-yl)-1-(pyrazin-2-yl)-4,5 ,6,7-Tetrahydro-1H-indazole-3-carboxamide ( 5h )

(S)-7,7-difluoro-N-(1-hydroxy-3,3-dimethylbutan-2-yl)-1-(pyrazin-2-yl)-4,5,6,7-tetrahydro-1H- indazole-3-carboxamide

Compound 5g (5.0g, 32.5mmol) was dissolved in dichloromethane (10mL), HATU (2M, 10mL), TEA (2M, 10mL), (S)-tertiary leucinol (3.97 mL) were added thereto at room temperature g, 39 mmol). The reaction was carried out at room temperature for two hours. Dilute hydrochloric acid was adjusted to neutrality, the aqueous phase was extracted with dichloromethane (100 mL×2), dried over sodium sulfate, and separated by column chromatography (eluent: petroleum ether/ethyl acetate=2:1~1:1) to obtain compound 5h (0.1 g, 52%).

LC-MS (ESI): m/z=380.2 [M+H] ⁺ .

The eighth step: (S)-3-(7,7-difluoro-3-((1-hydroxy-3,3-dimethylbutan-2-yl)aminocarboxy)-4,5, 6,7-Tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide ( Compound 5 )

(S)-3-(7,7-difluoro-3-((1-hydroxy-3,3-dimethylbutan-2-yl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1- yl)pyrazine 1-oxide

The raw material 5h (100mg, 0.35mmol) was dissolved in 5ml of formic acid solution, then 30% hydrogen peroxide solution (0.5ml) was added, the temperature was raised to 64°C and stirred for 3 hours, then 50ml of water was added, and 50ml of ethyl acetate was added. Extract three times, combine the organic phases, dry over anhydrous sodium sulfate, and dissolve the obtained brown oil in a mixed solvent of 3 ml of methanol and 3 ml of tetrahydrofuran after spin-drying, then slowly dropwise add 2 ml (2M) of an aqueous solution of sodium hydroxide, After 15 minutes of stirring reaction at normal temperature, TLC detected, the reaction was complete, 50ml of water was added, extracted three times with 50ml of ethyl acetate, the combined organic phases were dried over anhydrous sodium sulfate, dried and spin-dried, and the final product white powder 32mg ( Compound 5 ), yield 26%.

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. Peak time: 13.8min. LC-MS (ESI): m/z=396.2 [M+H] ⁺ .

¹ H NMR (400MHz, Chloroform-d)δ 8.70(dd,1H), 8.40(dd,1H), 8.08(dd,1H), 7.04(d,1H), 4.06-3.87(m,2H), 3.69( dd, 1H), 3.00 (dd, 2H), 2.38-2.14 (m, 2H), 2.10-1.99 (m, 3H), 1.04 (s, 9H).

Example 6

(S)-3-(3'-((1-Hydroxy-3,3-dimethylbutan-2-yl)aminocarboxy)-5',6'-dihydrospiro[cyclopropyl- 1,7'-Indazol]-1'(4'H)-yl)pyrazine 1-oxide (Compound 6 )

(S)-3-(3'-((1-hydroxy-3,3-dimethylbutan-2-yl)carbamoyl)-5',6'-dihydrospiro[cyclopropane-1,7'-indazol]-1'( 4'H)-yl)pyrazine 1-oxide

The first step: 2-(2-chloroethyl)cyclohexanone ( 6b )

2-(2-chloroethyl)cyclohexanone

At room temperature, acetonitrile (3.3 mL) was added to compound 6a (3.3 g, 23.24 mmol), concentrated hydrochloric acid (16.5 ml) was added at 0 °C, the temperature was raised to 100 °C and reacted for 3 hours, the reaction solution was cooled to room temperature and then washed with acetic acid. After extraction with ethyl ester, the organic phase was concentrated to dryness to obtain compound 6b (3.68 g, yield 99%).

The second step: spiro[2.5]octan-4-one ( 6c ).

spiro[2.5]octan-4-one.

At room temperature, ethanol (16 mL) and potassium hydroxide (1.8 g, 32 mmol) were added to compound 6b (3.65 g, 23 mmol), and the reaction was carried out at room temperature for 5 hours. The reaction solution was poured into (50 mL) water, extracted with ethyl acetate (50 mL×2), the organic phase was concentrated, and the residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate (v/v)=0:1~1 : 8) to obtain compound 6c (1 g, yield 35%).

The third step: methyl 2-oxo-2-(4-oxospiro[2.5]oct-5-yl)acetate ( 6d )

methyl 2-oxo-2-(4-oxospiro[2.5]octan-5-yl)acetate

Compound 6c (500 mg, 4 mmol) was added to tetrahydrofuran (10 mL), bistrimethylsilylamide lithium (5 ml, 5 mmol) was added at -78 °C, stirred at -78 °C for 30 min, and 2-chloro-2-oxo was added Methyl acetate (740 mg, 6.1 mmol), continued to react for 1 h, slowly warmed to room temperature, the reaction solution was quenched with aqueous ammonium chloride solution, poured into water (20 mL), extracted with ethyl acetate (30 mL×3), the organic phase was After concentration, the residue was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate (v/v)=0:1~1:6) to obtain compound 6d (700 mg, yield 83%).

LC-MS (ESI): m/z=211.1 [M+H] ⁺ .

The fourth step: 1'-(pyrazin-2-yl)-1',4',5',6'-tetrahydrospiro[cyclopropane-1,7'-indazole]-3'-carboxylate Esters ( 6e )

methyl 1'-(pyrazin-2-yl)-1',4',5',6'-tetrahydrospiro[cyclopropane-1,7'-indazole]-3'-carboxylate

At room temperature, methanol (6 mL) and 2,4-difluorophenylhydrazine (252 mg, 2.3 mmol) were sequentially added to compound 6d (400 mg, 1.9 mmol), and the pH value was adjusted to 4-5 with hydrochloric acid (6 M), and heated to The reaction was stirred at 50°C for 2 hours. The reaction solution was poured into (30 mL) water, extracted with ethyl acetate (20 mL×2), the organic phase was concentrated, and the residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate (v/v)=0:1~ 1:3) to obtain compound 6e (300 mg, yield 56%).

LC-MS (ESI): m/z=285.1 [M+H] ⁺ .

The fifth step: 1'-(pyrazin-2-yl)-1',4',5',6'-tetrahydrospiro[cyclopropane-1,7'-indazole]-3'-carboxylic acid ( 6f )

1'-(pyrazin-2-yl)-1',4',5',6'-tetrahydrospiro[cyclopropane-1,7'-indazole]-3'-carboxylic acid

At room temperature, methanol (3 ml) and water (1 ml) were added to compound 6e (180 mg, 0.63 mmol), and then sodium hydroxide (120 mg, 3 mmol) was added, and the reaction was stirred for 1 hour. The reaction solution was adjusted to pH 4-5 with dilute hydrochloric acid, extracted with ethyl acetate (20 mL×2), and the organic phase was concentrated to obtain compound 6f (165 mg, yield 97%).

LC-MS (ESI): m/z=271.1 [M+H] ⁺ .

The sixth step: 3-(3'-carboxy-5',6'-dihydrospiro[cyclopropane-1,7'-indazole]-1'(4'H)-yl)pyrazine 1-oxide ( 6g )

3-(3'-carboxy-5',6'-dihydrospiro[cyclopropane-1,7'-indazol]-1'(4'H)-yl)pyrazine 1-oxide

At room temperature, to compound 6f (165 mg, 0.61 mmol), formic acid (5 mL) and 30% hydrogen peroxide (0.5 mL) were sequentially added, and the reaction was heated to 50° C. and stirred for 2 hours. The reaction solution was poured into (20 mL) water, extracted with ethyl acetate (20 mL×2), and the organic phase was concentrated to obtain compound 6g (140 mg, yield 80%).

LC-MS (ESI): m/z=287.1 [M+H] ⁺ .

The seventh step: (S)-3-(3'-((1-hydroxy-3,3-dimethylbutan-2-yl)aminocarboxy)-5',6'-dihydrospiro[ Cyclopropane-1,7'-indazol]-1'(4'H)-yl)pyrazine 1-oxide ( Compound 6 )

(S)-3-(3'-((1-hydroxy-3,3-dimethylbutan-2-yl)carbamoyl)-5',6'-dihydrospiro[cyclopropane-1,7'-indazol]-1'( 4'H)-yl)pyrazine 1-oxide

At room temperature, to compound 6g (140mg, 0.49mmol) were sequentially added DMF (5mL), S-tertiary leucinol (60mg, 0.0.51mmol), HATU (223mg, 0.0.58mmol), DIPEA (160mg, 1.2mmol) ). The reaction was stirred for 1 hour. The reaction solution was poured into (50 mL) water, extracted with ethyl acetate (20 mL×2), the organic phase was concentrated, and separated and purified by preparative HPLC to obtain compound 6 (35 mg, yield 19%).

Preparative HPLC separation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: mobile phase A: acetonitrile; mobile phase B: water (containing 0.1% ammonium acetate); b. gradient elution, mobile phase A content from 5% to 50% ; c. flow rate 12mL/min; d flushing time 20min. Compound 6 residence time: 13.5 min.

LC-MS (ESI): m/z=386.2 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ ) δ 8.49(s, 1H), 8.27(d, 1H), 8.13-8.07(m, 1H), 7.03(d, 1H), 4.03-3.94(m, 2H), 3.7 -3.65(m,1H),2.98(t,2H),2.41(s,1H),1.95-1.88(m,2H),1.64-1.61(m,2H),1.41-1.38(m,2H),1.03 (s, 9H), 0.71-0.68 (m, 2H).

Example 7

3-((R)-3-((S)-1-Hydroxy-3,3-dimethylbutan-2-yl)aminocarboxy-4-methyl-7-(propan-2-ylidene )-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide (Compound 7)

3-((R)-3-(((S)-1-hydroxy-3,3-dimethylbutan-2-yl)carbamoyl)-4-methyl-7-(propan-2-ylidene)-4,5, 6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide.

The first step: methyl 2-((6R)-6-methyl-2-oxo-3-(propane-2-ylidene)cyclohexyl)-2-oxoacetate ( 7b )

methyl 2-((6R)-6-methyl-2-oxo-3-(propan-2-ylidene)cyclohexyl)-2-oxoacetate

Under nitrogen protection, tetrahydrofuran (10 mL) was added to compound 7a (0.5 g, 3.3 mmol), cooled to -70 to -80 °C, lithium bistrimethylsilylamide (4.3 mL, 4.3 mmol) was added to -70 Add dropwise to the reaction solution at a temperature of -80 °C, keep the reaction for half an hour, add oxalyl chloride monomethyl ester (0.5g, 4.3 mmol) dropwise to the reaction solution at -70 to -80 °C, keep the reaction for half an hour . The reaction solution was poured into (50 mL) water, adjusted to pH 5-6 with hydrochloric acid (6M), extracted with ethyl acetate (30 mL×2), the organic phase was concentrated, and subjected to column chromatography (eluent PE/EA=10/1). -3/1) separation and purification to obtain compound 7b (0.3 g, yield 38%).

LC-MS (ESI): m/z=239.1 [M+H] ⁺ .

The second step: (R)-3-(3-(methoxycarbonyl)-4-methyl-7-(propane-2-ylidene)-4,5,6,7-tetrahydro-1H-indole oxazol-1-yl)pyrazine 1-oxide ( 7c )

(R)-3-(3-(methoxycarbonyl)-4-methyl-7-(propan-2-ylidene)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

At room temperature, methanol (5 mL) and pyrazine-2-hydrazine oxide (0.2 g, 1.6 mmol) were sequentially added to compound 7b (0.3 g, 1.3 mmol), and the pH value was adjusted to 3-4 with hydrochloric acid (6 M), Heat to 50°C and stir the reaction for 2 hours. The reaction solution was poured into (30 mL) water, extracted with ethyl acetate (20 mL×2), the organic phase was concentrated, and separated and purified by column chromatography (eluent PE/EA=5/1-1/1) to obtain compound 7c (0.2 g, 50% yield).

LC-MS (ESI): m/z=329.1 [M+H] ⁺ .

The third step: (R)-3-(3-carboxy-4-methyl-7-(propane-2-ylidene)-4,5,6,7-tetrahydro-1H-indazol-1-yl ) pyrazine 1-oxide ( 7d )

(R)-3-(3-carboxy-4-methyl-7-(propan-2-ylidene)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

MeOH (2 mL), water (2 mL), and sodium hydroxide (73.1 mg, 1.8 mmol) were sequentially added to compound 7c (0.2 g, 0.6 mmol) at room temperature, and the reaction was stirred for 1 hour. The reaction solution was poured into (10 mL) water, adjusted to pH 3-4 with hydrochloric acid (6M), extracted with ethyl acetate (10 mL×2), and the organic phase was concentrated to obtain compound 7d (150 mg, yield 80%).

LC-MS (ESI): m/z=315.1 [M+H] ⁺ .

The fourth step: 3-((R)-3-((S)-1-hydroxy-3,3-dimethylbutan-2-yl)amine carboxamide-4-methyl-7-(propane- 2-ylidene)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide ( Compound 7 )

3-((R)-3-(((S)-1-hydroxy-3,3-dimethylbutan-2-yl)carbamoyl)-4-methyl-7-(propan-2-ylidene)-4,5, 6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

At room temperature, to compound 7d (150 mg, 0.48 mmol) were sequentially added DMF (5 mL), S-tert-leucinol (62 mg, 0.53 mmol), HATU (200 mg, 0.53 mmol), DIPEA (185 mg, 1.44 mmol). The reaction was stirred for 1 hour. The reaction solution was poured into (50 mL) water, extracted with ethyl acetate (20 mL×2), the organic phase was concentrated, and separated and purified by silica gel column chromatography (eluent PE/EA=5/1-1/1) to obtain compound 7 (50 mg, 25% yield).

LC-MS (ESI): m/z=414.2 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ )δ 8.60(s, 1H), 8.31-8.30(d, 1H), 8.04-8.02(m, 1H), 7.10-7.08(d, 1H), 4.03-3.95(m, 2H), 3.69-3.65(m, 1H), 3.45-3.41(m, 1H), 2.54-2.51(m, 2H), 2.05-2.00(m, 1H), 1.81(s, 3H), 1.72-1.65( m, 1H), 1.35 (s, 3H), 1.30-1.28 (d, 3H), 1.03 (s, 9H).

Example 8

3-(7,7-Difluoro-3-((1-(hydroxymethyl)cyclopentyl)aminocarbamoyl)-4,5,6,7-tetrahydro-1H-indazole-1- yl)pyrazine 1-oxide (compound 8)

3-(7,7-difluoro-3-((1-(hydroxymethyl)cyclopentyl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

The first step: 7,7-difluoro-N-(1-(hydroxymethyl)cyclopentyl)-1-(pyrazin-2-yl)-4,5,6,7-tetrahydro-1H- Indazole-3-carboxamide ( 8b )

7,7-difluoro-N-(1-(hydroxymethyl)cyclopentyl)-1-(pyrazin-2-yl)-4,5,6,7-tetrahydro-1H-indazole-3-carboxamide

Referring to the seventh step of compound 5 , compound 8b is obtained by reacting compound 5g .

The second step: (S)-3-(7,7-difluoro-3-((1-hydroxy-3,3-dimethylbutan-2-yl)aminocarboxy)-4,5, 6,7-Tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide ( Compound 8 )

(S)-3-(7,7-difluoro-3-((1-hydroxy-3,3-dimethylbutan-2-yl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1- yl)pyrazine 1-oxide

Referring to the eighth step of compound 5 , compound 8 (30 mg, 32%) was obtained by the reaction of compound 8b .

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. Peak time: 12.8min.

LC-MS (ESI): m/z=394.1 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ )δ 8.75(t,1H), 8.43(d,1H), 8.11(dd,1H), 7.10(s,1H), 3.79(s,2H), 3.27(s,1H) ), 2.99(t, 2H), 2.37-2.22(m, 2H), 2.12-1.96(m, 4H), 1.96-1.66(m, 6H).

Example 9

3-(7,7-Difluoro-3-((4,4,4-trifluoro-1-hydroxy-3,3-dimethylbutan-2-yl)aminocarboxy)-4,5, 6,7-Tetrahydro-1H-indazol-1-yl)pyrazine-1-oxide

3-(7,7-difluoro-3-((4,4,4-trifluoro-1-hydroxy-3,3-dimethylbutan-2-yl)carbamoyl)-4,5,6,7-tetrahydro-1H- indazol-1-yl)pyrazine-1-oxide

The first step: 3-(7,7-difluoro-3-((4,4,4-trifluoro-1-hydroxy-3,3-dimethylbutan-2-yl)aminoformamide)- 4,5,6,7-Tetrahydro-1H-indazol-1-yl)oxopyrazine-1-oxide

3-(7,7-difluoro-3-((4,4,4-trifluoro-1-hydroxy-3,3-dimethylbutan-2-yl)carbamoyl)-4,5,6,7-tetrahydro-1H- indazol-1-yl)pyrazine-1-oxide

Under nitrogen protection, to compound 10a (0.05 g, 0.17 mmol), DMF (3 mL), DIPEA (32 mg, 0.25 mmol), HATU (77 mg, 0.2 mmol), 2-amino-4,4 were sequentially added at room temperature , 4-Trifluoro-3,3-dimethylbutan-1-ol (Int-1) (43 mg, 0.25 mmol), and stirred at room temperature for 3 h. Saturated aqueous sodium bicarbonate solution (30 mL) was added to the reaction to quench the reaction, the aqueous phase was extracted with ethyl acetate (20 mL×2), the layers were left to stand, the aqueous phase was extracted with dichloromethane (20 mL), and the combined organic phases were It was dried over anhydrous sodium sulfate, and the residue was concentrated under reduced pressure to obtain a crude product. The crude product was separated by HPLC to obtain compound 9 (30 mg, yield 39.64%) as a white solid.

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. Peak time: 12.5min. LC-MS (ESI): m/z=450.2 [M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ )δ 9.14(s, 1H), 8.53(s, 1H), 8.41(d, J =8.0Hz, 1H), 8.13(d, J =8.0Hz, 1H), 4.81( s,1H),4.34-4.29(m,2H),4.08-3.70(m,1H),3.63-3.51(m,1H),3.31-3.17(m,1H),2.90-2.73(m,2H), 2.34-2.27(m, 2H), 1.24-1.17(s, 6H).

Example 10:

(S)-3-(7,7-Difluoro-3-((2-hydroxy-1-phenethyl)aminocarboxy)-4,5,6,7-tetrahydro-1H-indazole -1-yl)pyrazine 1-oxide (compound 10)

(S)-3-(7,7-difluoro-3-((2-hydroxy-1-phenylethyl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

The first step: 3-(3-carboxy-7,7-difluoro-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide ( 10a )

3-(3-carboxy-7,7-difluoro-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

The intermediate 5g (2g, 7.14mmol) was dissolved in formic acid (20ml), then 30% hydrogen peroxide solution (2ml) was added, the temperature was raised to 64 ° C and stirred for 3 hours, then 50ml of water was added, and cooled to room temperature, reduced Press suction filtration to obtain compound 10a (800 mg, 38%).

LC-MS (ESI): m/z=297.3 [M+H] ⁺ .

The second step: (S)-3-(7,7-difluoro-3-((2-hydroxy-1-phenethyl)aminocarbamoyl)-4,5,6,7-tetrahydro- 1H-Indazol-1-yl)pyrazine 1-oxide ( Compound 10 )

(S)-3-(7,7-difluoro-3-((2-hydroxy-1-phenylethyl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

Compound 10a (200 mg, 0.68 mmol) was dissolved in dichloromethane (10 mL), to which was added HATU (334 mg, 0.88 mL), DIPEA (2.0 mL), (S)-2-amino-2- Phenylethane-1-ol (102 mg, 0.75 mmol). The reaction was carried out at room temperature for two hours. Dilute hydrochloric acid was adjusted to neutrality, the aqueous phase was extracted with dichloromethane (20 mL×2), the organic phase was dried with sodium sulfate, and the organic phase was dried by spin drying and separated and purified by high-performance liquid phase preparation. Compound 10 was obtained (0.11 g, 38%).

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. Peak time: 13.2min.

LC-MS (ESI): m/z=416.4 [M+H] ⁺ .

¹ H NMR (400MHz, Chloroform-d)δ 8.76(dd,1H), 8.38(dd,1H), 8.06(dd,1H), 7.60(d,1H), 7.39(d,3H), 7.35-7.28( m, 1H), 5.25 (t, 1H), 4.01 (d, 2H), 2.97 (tt, 2H), 2.27 (t, 2H), 2.16-1.84 (m, 3H).

Example 11:

3-(7,7-Difluoro-3-((2-phenylpropan-2-yl)aminocarboxy)-4,5,6,7-tetrahydro-1H-indazol-1-yl) Pyrazine 1-oxide (Compound 11)

3-(7,7-difluoro-3-((2-phenylpropan-2-yl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

Compound 10a (200 mg, 0.68 mmol) was dissolved in dichloromethane (10 mL), and thereto were added HATU (334 mg, 0.88 mL), DIPEA (2.0 mL), 2-phenylpropan-2-amine (101 mg) at room temperature , 0.75mmol). The reaction was carried out at room temperature for two hours. Dilute hydrochloric acid was adjusted to neutrality, the aqueous phase was extracted with dichloromethane (20 mL×2), the organic phase was dried with sodium sulfate, and the organic phase was dried by spin drying and separated and purified by high-performance liquid phase preparation. Compound 11 was obtained (0.03 g, 11%).

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. Peak time: 13.8min.

LC-MS (ESI): m/z=414.3 [M+H] ⁺ .

¹ H NMR (400MHz, Chloroform-d) δ 8.72 (dd, 1H), 8.40 (dd, 1H), 8.07 (dd, 1H), 7.49-7.41 (m, 2H), 7.35 (dd, 2H), 7.25- 7.17(m, 1H), 2.93(tt, 2H), 2.25(t, 2H), 2.04-1.95(m, 2H), 1.82(s, 6H), 1.66(s, 1H).

Example 12:

3-(7,7-Difluoro-3-((2-methyl-1-morpholinopropan-2-yl)aminocarbamoyl)-4,5,6,7-tetrahydro-1H- Indazol-1-yl)pyrazine 1-oxide (Compound 12)

3-(7,7-difluoro-3-((2-methyl-1-morpholinopropan-2-yl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

Compound 10a (200 mg, 0.68 mmol) was dissolved in dichloromethane (10 mL), and thereto were added HATU (334 mg, 0.88 mmol), DIPEA (2.0 mL), 2-methyl-1-morpholinopropane at room temperature -2-amine (215 mg, 1.36 mmol). The reaction was carried out at room temperature for two hours. Dilute hydrochloric acid was adjusted to neutrality, the aqueous phase was extracted with dichloromethane (20 mL×2), the organic phase was dried over anhydrous sodium sulfate, the solvent was removed under reduced pressure, and high-performance liquid phase was prepared for separation and purification. Compound 12 was obtained (20 mg, 5%).

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. out of the peak Time: 13.7min.

LC-MS (ESI): m/z=437.3 [M+H] ⁺ .

¹ H NMR (400MHz, Chloroform-d)δ 8.65(s, 1H), 8.38(d, 1H), 8.06(dd, 1H), 7.32(s, 1H), 3.71(s, 4H), 2.98(s, 2H), 2.61 (d, 4H), 2.27 (t, 2H), 2.05 (t, 2H), 1.46 (s, 6H).

Example 13:

3-(7,7-Difluoro-3-((1-methoxy-2-methyl-1-oxopropan-2-yl)aminocarbamoyl)-4,5,6,7-tetra Hydro-1H-indazol-1-yl)pyrazine 1-oxide (Compound 13)

3-(7,7-difluoro-3-((1-methoxy-2-methyl-1-oxopropan-2-yl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl) pyrazine 1-oxide

Compound 10a (100 mg, 0.34 mmol) was dissolved in dichloromethane (5 mL), to which was added HATU (194 mg, 0.51 mmol), DIPEA (0.2 mL), 2-amino-2-methylpropionic acid at room temperature Methyl ester (40 mg, 0.34 mmol). The reaction was carried out at room temperature for two hours. Dilute hydrochloric acid was adjusted to neutrality, the aqueous phase was extracted with dichloromethane ( 20 mL×2), the organic phase was dried over sodium sulfate, and the solvent was removed under reduced pressure.

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. Peak time: 12.9min.

LC-MS (ESI): m/z=396.1 [M+H] ⁺ .

¹ H NMR (400MHz, Chloroform-d)δ 8.73(s,1H), 8.40(d,1H), 8.08(dd,1H), 7.36(s,1H), 3.78(s,3H), 2.97(t, 2H), 2.39-2.13 (m, 2H), 2.11-1.90 (m, 2H), 1.67 (s, 6H).

Example 14:

3-(3-((2-Cyanopropan-2-yl)aminocarbamoyl)-7,7-difluoro-4,5,6,7-tetrahydro-1H-indazol-1-yl ) pyrazine 1-oxide (compound 14)

3-(3-((2-cyanopropan-2-yl)carbamoyl)-7,7-difluoro-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

Compound 10a (100 mg, 0.34 mmol) was dissolved in dichloromethane (5 mL), to which was added HATU (194 mg, 0.51 mmol), DIPEA (0.2 mL), 2-amino-2-methylpropionitrile at room temperature (29 mg, 0.34 mmol). The reaction was carried out at room temperature for two hours. Dilute hydrochloric acid was adjusted to neutrality, the aqueous phase was extracted with dichloromethane (20 mL×2), the organic phase was dried over sodium sulfate, the solvent was removed under reduced pressure, and high-performance liquid phase preparation was separated and purified to obtain compound 14 (10 mg, 7%).

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. Peak time: 12.6min.

LC-MS (ESI): m/z=363.1 [M+H] ⁺ .

¹ H NMR (400MHz, Chloroform-d) δ 8.68(dd, 1H), 8.48-8.35(m, 1H), 8.09(dd, 1H), 6.92(s, 1H), 3.01(t, 2H), 2.42- 2.17(m, 2H), 2.15-1.96(m, 2H), 1.58(s, 6H).

Example 15:

3-(3-((2-Cyanopropan-2-yl)aminocarbamoyl)-7,7-difluoro-4,5,6,7-tetrahydro-1H-indole Azol-1-yl)pyrazine 1-oxide (Compound 15)

3-(3-((2-cyanopropan-2-yl)carbamoyl)-7,7-difluoro-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

Compound 10a (100 mg, 0.34 mmol) was dissolved in dichloromethane (5 mL), to which was added HATU (194 mg, 0.51 mmol), DIPEA (0.2 mL), (tetrahydro-2H-pyran-4-) at room temperature yl)methylamine (39 mg, 0.34 mmol). The reaction was carried out at room temperature for two hours. Dilute hydrochloric acid was adjusted to neutrality, dichloromethane (20 mL×2) was used to extract the aqueous phase, the organic phase was dried with sodium sulfate, the solvent was removed under reduced pressure, and high-performance liquid phase preparation was separated and purified to obtain compound 15 (10 mg, 7%).

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. Peak time: 12.5min.

LC-MS (ESI): m/z=394.1 [M+H] ⁺ .

¹ H NMR (400MHz, Chloroform-d) δ 8.69(s, 1H), 8.40(d, 1H), 8.07(dd, 1H), 6.98(s, 1H), 4.00(dd, 2H), 3.46-3.29( m, 2H), 2.99 (t, 2H), 2.39-2.17 (m, 2H), 2.13-1.95 (m, 2H), 1.68 (d, 8H), 1.39 (t, 2H).

Example 16:

3-(7,7-Difluoro-3-((tetrahydro-2H-pyran-4-yl)aminocarboxy)-4,5,6,7-tetrahydro-1H-indazole-1 -yl)pyrazine 1-oxide (compound 16)

3-(7,7-difluoro-3-((tetrahydro-2H-pyran-4-yl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

Compound 10a (100 mg, 0.34 mmol) was dissolved in dichloromethane (5 mL), to which was added HATU (194 mg, 0.51 mmol), DIPEA (0.2 mL), tetrahydro-2H-pyran-4-amine at room temperature (34 mg, 0.34 mmol). The reaction was carried out at room temperature for two hours. Dilute hydrochloric acid was adjusted to neutrality, dichloromethane (20 mL×2) was used to extract the aqueous phase, the organic phase was dried with sodium sulfate, the solvent was removed under reduced pressure, and high-performance liquid phase preparation was separated and purified to obtain compound 16 (20 mg, 15%).

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. Peak time: 12.3min.

LC-MS (ESI): m/z=380.1 [M+H] ⁺ .

¹ H NMR (400MHz, Chloroform-d)δ 8.72(dd,1H), 8.40(dd,1H), 8.08(dd,1H), 6.80(d,1H), 4.02(dd,2H), 3.53(t, 2H), 3.00 (tq, 2H), 2.36-2.21 (m, 2H), 1.98 (t, 1H), 1.63 (t, 6H).

Example 17:

3-(7,7-Difluoro-3-((2-(hydroxymethyl)oxetan-2-yl)aminocarbamoyl)-4,5,6,7-tetrahydro-1H- Indazol-1-yl)pyrazine 1-oxide (Compound 17)

3-(7,7-difluoro-3-((2-(hydroxymethyl)oxetan-2-yl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

Compound 10a (100 mg, 0.34 mmol) was dissolved in dichloromethane (5 mL), HATU (194 mg, 0.51 mmol), DIPEA (0.2 mL), (2-aminooxetane-2) were added thereto at room temperature -yl) methanol (35 mg, 0.34 mmol). The reaction was carried out at room temperature for two hours. Dilute hydrochloric acid was adjusted to neutrality, the aqueous phase was extracted with dichloromethane (20 mL×2), the organic phase was dried over sodium sulfate, the solvent was removed under reduced pressure, and high-performance liquid phase preparation was separated and purified to obtain compound 17 (34 mg, 22%).

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. Peak time: 12.4min.

LC-MS (ESI): m/z=382.1 [M+H] ⁺ .

¹ H NMR (400MHz, Chloroform-d)δ 8.73(d,1H), 8.41(d,1H), 8.09(t,1H), 7.44(s,1H), 4.88-4.81(m,2H), 4.69( d, 2H), 4.17 (s, 2H), 3.01-2.89 (m, 2H), 2.35-2.16 (m, 2H), 2.13-1.94 (m, 2H).

Example 18:

3-(7,7-Difluoro-3-((1-phenylcyclopropyl)aminocarboxy)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyridine Azine 1-oxide (Compound 18)

3-(7,7-difluoro-3-((1-phenylcyclopropyl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

Compound 10a (100 mg, 0.34 mmol) was dissolved in dichloromethane (5 mL), to which were added HATU (194 mg, 0.51 mmol), DIPEA (0.2 mL), 1-phenylcyclopropan-1-amine ( 45 mg, 0.34 mmol). The reaction was carried out at room temperature for two hours. Dilute hydrochloric acid was adjusted to neutrality, dichloromethane (20 mL×2) was used to extract the aqueous phase, the organic phase was dried over sodium sulfate, the solvent was removed under reduced pressure, and high-performance liquid phase preparation was separated and purified to obtain compound 18 (33 mg, 24%).

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. Peak time: 13.1min.

LC-MS (ESI): m/z=412.1 [M+H] ⁺ .

¹ H NMR (400MHz, Chloroform-d)δ 8.79(d,1H), 8.37(d,1H), 8.05(dd,1H), 7.67(s,1H), 7.37-7.24(m,4H), 7.24- 7.15(m, 1H), 2.98(t, 2H), 2.27(t, 2H), 2.07-1.99(m, 2H), 1.38(t, 2H).

Example 19:

3-(7,7-Difluoro-3-((1-phenylcyclopropyl)aminocarboxy)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyridine Azine 1-oxide (Compound 19)

3-(7,7-difluoro-3-((1-phenylcyclopropyl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

Compound 10a (100 mg, 0.34 mmol) was dissolved in dichloromethane (5 mL), to which was added HATU (194 mg, 0.51 mmol), DIPEA (0.2 mL), (S)-3-aminopropane-1 at room temperature , 2-diol (31 mg, 0.34 mmol). The reaction was carried out at room temperature for two hours. Dilute hydrochloric acid was adjusted to neutrality, the aqueous phase was extracted with dichloromethane (20 mL×2), the organic phase was dried over sodium sulfate, the solvent was removed under reduced pressure, and high-performance liquid phase preparation was separated and purified to obtain compound 19 (42 mg, 36%).

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire @ Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. Peak time: 13.3min.

LC-MS (ESI): m/z=370.1 [M+H] ⁺ .

¹ H NMR (400MHz, Chloroform-d) δ 8.72(s, 1H), 8.40(d, 1H), 8.13-8.04(m, 1H), 7.37(s, 1H), 3.97-3.84(m, 1H), 3.75-3.53(m, 2H), 2.98(s, 2H), 2.36-2.19(m, 2H), 2.12-1.95(m, 2H), 1.26(s, 2H).

Example 20:

3-(7,7-Difluoro-3-((1-phenylcyclopropyl)aminocarboxy)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyridine oxazine 1-oxide (compound 20)

3-(7,7-difluoro-3-((1-phenylcyclopropyl)carbamoyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl)pyrazine 1-oxide

Compound 10a (100 mg, 0.34 mmol) was dissolved in dichloromethane (5 mL), to which was added HATU (194 mg, 0.51 mmol), DIPEA (0.2 mL), (R)-2-amino-4- at room temperature Methylpentan-1-ol (40 mg, 0.34 mmol). The reaction was carried out at room temperature for two hours. Dilute hydrochloric acid was adjusted to neutrality, the aqueous phase was extracted with dichloromethane (20 mL×2), the organic phase was dried over sodium sulfate, the solvent was removed under reduced pressure, and high-performance liquid phase preparation was separated and purified to obtain compound 20 (24 mg, 18%).

Preparation method: 1. Instrument: waters 2767 preparative liquid phase; chromatographic column: SunFire@Prep C18 (19mm×250mm). 2. The sample was dissolved in DMF and filtered with a 0.45 μm filter to prepare a sample solution. 3. Preparative chromatography conditions: a. Composition of mobile phases A and B: Mobile phase A: acetonitrile, mobile phase B: water; b. Gradient elution, mobile phase A content from 20% to 75%; c. Flow rate 12ml/min ; d. The brewing time is 20min. Peak time: 13.5min.

LC-MS (ESI): m/z=396.1 [M+H] ⁺ .

¹ H NMR (400MHz, Chloroform-d)δ 8.74(s, 1H), 8.39(d, 1H), 8.08(dd, 1H), 6.99(d, 1H), 4.24(t, 1H), 3.79(dd, 1H), 3.66(dd, 1H), 2.99(d, 2H), 2.34-2.20(m, 2H), 2.14-1.90(m, 4H), 1.71(d, 1H), 1.51(t, 2H), 0.97 (dd, 6H).

biological test

1. CB2 agonist activity

Test Items: Compounds 1-20.

CHO cells with high expression of human CB2 were suspended in HBSS buffer (containing 20mM HEPES, pH=7.4), and 7.5×10 ³ cells per well (the cell density was 1.5×10 ⁶ cells/mL) were seeded on the added sample (HBSS). , positive control WIN 55212-2 or different concentrations of example compounds) in 384-well white plates. A final concentration of 3 μM adenylate cyclase activator NKH 477 was added to the above 384-well plate. After 10 min incubation at 37°C, cells were lysed and D2-labeled cAMP and anti-cAMP antibody labeled with europium cryptate were added. After 1 hour incubation at room temperature, HTRF detection (λex=337 nm, λem=620 and 665 nm) was performed with a microplate reader (Envison, Perkin Elmer). The signal ratio Ratio (Signal 665/Signal 620*10 ⁴ ) was calculated, and the results were expressed as a percentage value relative to 100 nM WIN 55212-2, and the _EC50 value was fitted using the DoseResp function. The results are shown in Table 1.

APD-371 is compound 699 of patent document WO2011025541.

Conclusion: The compounds of the present invention, especially 1, 4, 5, 6, 7, 8, 19, and 20, showed higher agonistic activity for CB ₂ receptors, and some compounds, especially compound 5 and compound 8, showed high agonistic activity to CB ₂ /CB ₁ receptor showed higher selectivity.

2. Pharmacokinetic test in rats

Experimental purpose: The test substance was administered to SD rats through a single dose of intravenous and intragastric administration, the concentration of the test substance in the rat plasma was determined, and the pharmacokinetic characteristics and bioavailability of the test substance in rats were evaluated.

Test objects: APD-371 and compound 5.

Test animals: male SD rats, about 220 g, 6-8 weeks old, 6 rats/compound. Purchased from Chengdu Dashuo Laboratory Animal Co., Ltd.

Test method: On the test day, 6 SD rats were randomly divided into groups according to their body weight. Fasting for 12-14 hours on the day before administration, and 4 hours after administration. Dosing according to Table 2.

sampling

Before and after administration of isoflurane anesthesia, 0.1 ml of blood was collected from the orbit and placed in an EDTAK2 centrifuge tube. The plasma was collected by centrifugation at 5000 rpm and 4°C for 10 min.

Time points of plasma collection in group G1: 0, 2, 5, 15, 30 min, 1, 2, 4, 6, 8, 24 h.

Time points of plasma collection in group G2: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 24 h.

All samples were stored at -80°C prior to analysis.

sample preparation

Take 30 μL of plasma sample, standard curve and quality control sample, add 200 μL of acetonitrile solution containing internal standard, vortex and mix, and then centrifuge at 12000 rpm for 10 min at 4°C. 170 μL of supernatant was taken into a 96-well plate for LC-MS/MS analysis, and the injection volume was 5 μL.

The main pharmacokinetic parameters were analyzed using WinNonlin 8.0 software non-compartmental model. The experimental results are shown in Table 3.

APD-371 is compound 699 of patent document WO2011025541.

Conclusion: The compounds of the present invention, especially compound 5, have high bioavailability, good pharmacokinetic characteristics and long half-life.

3. Stability test

experimental method:

Solution stability: (injection volume 20ul)

Artificial gastric juice: take 16.4ml of dilute hydrochloric acid, add about 800ml of water and 10g of pepsin, shake well, and dilute to 1000ml with water.

Artificial intestinal juice: potassium dihydrogen phosphate buffer solution: Weigh 1.3961 g of potassium dihydrogen phosphate, dissolve in water, dilute to 100 ml, and adjust the pH to 6.8 with 0.1 mol/L sodium hydroxide solution.

Another take 2.0307g of trypsin, add water to dissolve, mix with potassium dihydrogen phosphate buffer solution, add water to 200ml, and shake well.

Take an appropriate amount of this product APD-371, add artificial gastric juice to dilute it to make a test solution containing about 0.6 mg per ml, place it at room temperature, and take it at 0, 2, 4, 8, 12, 24, 36, and 48 hours respectively. An appropriate amount of the test solution was injected into the liquid chromatograph, and the stability of the test solution under artificial gastric juice was investigated by sample injection analysis. Calculate the sample stability in terms of relative 0h peak area, and pay attention to whether there are new impurities.

Take an appropriate amount of compound 5 of this product, add artificial intestinal juice to dilute to make a test solution containing about 0.6 mg per ml, place it at room temperature, and take the test solution at 0, 2, 4, 8, 12, 24, 36, and 48 hours, respectively. An appropriate amount of the product solution was injected into a liquid chromatograph, and the sample solution was analyzed to investigate the stability of the test solution under artificial intestinal juice. Calculate the sample stability in terms of relative 0h peak area, and pay attention to whether there are new impurities.

Chemical stability: (injection volume 2ul)

Appropriate amounts of APD-371 and compound 5 were taken, respectively, and were placed in a colorless and transparent flat weighing bottle respectively, and sealed. The sealed samples were placed under the conditions of light and 40°C + 75% RH, and were sampled for 7 days and 33 days respectively.

Take an appropriate amount of the sample under each condition, add acetonitrile to make a test solution of about 0.2 mg/ml, inject the sample and analyze it, check the stability of the sample by the percentage of peak area, and pay attention to whether there is any new impurity.

The experimental conditions are shown in Table 4, and the experimental results are shown in Table 5.

4. Rat tissue distribution experiment

4.1. Test animals: male SD rats, about 200 g, 6-8 weeks old, 12 rats/compound. Purchased from Chengdu Dashuo Laboratory Animal Co., Ltd.

4.2 Experimental scheme:

Test method: On the test day, 24 SD rats were randomly divided into groups according to their body weight. Fasting for 12-14 hours on the day before administration, and 4 hours after administration. Dosing according to Table 6.

Before and after administration of isoflurane anesthesia, 0.20 mL of blood was collected from the orbit, placed in an EDTAK2 centrifuge tube, centrifuged at 5000 rpm for 10 min at 4°C, and plasma was collected. Blood collection time point: 10min, 0.5, 1, 4h. All plasma samples were stored at -80°C prior to analytical testing.

Tissue (heart, liver, spleen, lung, kidney, brain, colon, rectum) sample collection time points after administration: 10min, 0.5, 1, 4h. After the animals were bled and euthanized, the tissues were collected, washed with normal saline, homogenized with 50% methanol at a ratio of m/v = 1:3, and stored in a -80° refrigerator for analysis.

4.3 The experimental results are shown in Table 7:

Conclusion: The tissue distribution results of compound 5 synthesized by the technology of the present invention show that the exposure in the brain is lower than that of APD-371.

5. The maximum tolerated dose (MTD) test of single oral gavage in SD rats

Test design:

A total of 50 rats, half male and half, divided into 5 groups, 5 rats/sex in each group, vehicle control group (20% SBE-β-CD), APD-371 low-dose group 300 mg/kg, APD-371 high-dose group 1000 mg/kg, compound 5 low-dose group 300 mg/kg, compound 5 high-dose group 1000 mg/kg, oral gavage, administration volume 10 mL/kg.

Results and conclusions:

APD-371 MTD was 300 mg/kg; Compound 5 MTD was 1000 mg/kg.

Claims (12)

A tetrahydroindazole compound represented by formula (I) and its stereoisomer, pharmaceutically acceptable salt, hydrate or N-oxide:

Wherein, R ¹ is selected from -L ₁ -L ₂ -L ₃ -L ₄ ; L ₁ is selected from C _1-6 alkylene, C _3-7 cycloalkenylene, C _3-7 cycloalkylene, C _3-6 unsaturated cycloalkylene group, C _3-6 heterocyclic group, C _6-10 arylene group, C _5-10 heteroarylene group, C _4-12 bridged ring, C _4-12 parallel ring, C _5-10 spiro rings or absent, the alkylene, cycloalkenylene, cycloalkylene, unsaturated cycloalkylene, heterocyclylene, arylene, heteroarylene, bridged ring, and The ring and spiro ring are each independently optionally substituted by the following substituents: halogen, cyano, hydroxyl, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, -C _2-6 alkynyl- C _3-6 cycloalkyl, C _1-6 alkoxy, hydroxy-substituted C _1-6 alkyl, halogenated C _1-6 alkyl, benzene ring, -C _1-6 alkyl-O-haloalkyl , -C(O)OC _1-6 alkyl or C _3-6 cycloalkyl; L ₂ is selected from C _1-6 alkylene, C _3-7 cycloalkylene, C _3-6 heterocyclylene , C _5-10 heteroarylene, -C(O)NH-, -C(O)-, -C(O)O- or absent, the alkylene, cycloalkylene, hetero The cyclic group and the heteroarylene group are each independently optionally substituted by the following substituents: C _1-6 alkyl or halogen; L ₃ is selected from C _1-6 alkylene or absent; L ₄ is selected from H, halogen, hydroxyl , amine group, ureido group, cyano group, C _1-6 alkyl group, C _3-7 cycloalkyl group, C _1-6 alkoxy group, C _3-6 heterocyclic group, C _2-6 alkynyl group, C _{6 -10} aryl, C _5-10 heteroaryl, -NHC _1-6 alkyl, -N(C _1-6 alkyl)C _1-6 alkyl, -NHC(O)C _1-6 alkyl, -C(O)NHC _1-6 alkyl, -NH-C _6-10 aryl, -NH-C _3-7 cycloalkyl, -C(O)-C _6-10 aryl, -OC _{6- 10} aryl, -OC _6-10 heteroaryl, -OC _1-6 alkylene-COOH, -C(O)NH ₂ , -COOH, C _4-12 bridged ring, C _4-12 cyclic ring or C _5-10 Spiro, the alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkynyl, bridged, spiro, and parallel are optionally substituted by the following substituents: halogen, cyano, hydroxyl, amino, C _1-6 alkyl, C _1-6 haloalkyl or C _1-6 alkoxy; R ² and R ³ are each independently selected from H, F, Cl, cyano, Hydroxyl or C _1-6 alkyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H, F, Cl, amino, cyano, hydroxyl, C _1-6 alkyl or C _1-6 haloalkane R ⁸ and R ⁹ are each independently selected from H, F, Cl, C _1-6 alkyl, and the alkyl is optionally substituted by the following substituents: cyano, F or Cl; or R ⁸ and R ⁹ together to form a C _2-6 alkenyl group or a C _3-6 ring Alkyl; R ¹⁰ is selected from H or C _1-6 alkyl; R ¹¹ is selected from

or

; the condition is: when R ¹¹ is selected from

, R ¹ is selected from

,

,

or

. The tetrahydroindazole compound and its stereoisomers, pharmaceutically acceptable salts, hydrates or N-oxides according to claim 1, wherein: R ¹ is selected from C _1-6 alkyl, C _{3- 6} cycloalkyl, C _3-6 heterocyclic group, C _3-6 unsaturated cyclic hydrocarbon group, C _6-10 aryl group, C _5-10 heteroaryl group, C _4-12 bridged ring, C _4-12 parallel ring , C _5-10 spiro ring, the alkyl, cycloalkyl, unsaturated cyclic hydrocarbon group, heterocyclic group, aryl, heteroaryl, bridged ring, and ring or spiro ring is optionally substituted by the following substituents: F, Cl, hydroxy, cyano, C _1-6 alkyl, hydroxy substituted C _1-6 alkyl, C _1-6 alkoxy, C _3-6 heterocyclyl, benzene ring, -C(O) OC _1-6 alkyl or C _3-6 cycloalkyl. The tetrahydroindazole compound and its stereoisomers, pharmaceutically acceptable salts, hydrates or N-oxides according to claim 2, wherein: R ¹ is selected from

,

,

,

, indolyl, pyrrolyl, pyridyl, oxidyl, phenyl, cyclohexyl, cyclopentyl, pyrazine, adamantyl, oxazine, morpholinyl,

,

,

,

,

or

; or R ¹ is selected from

,

,

,

,

,

. The tetrahydroindazole compound and its stereoisomers, pharmaceutically acceptable salts, hydrates or N-oxides according to claim 1, wherein: R ¹ is selected from C _1-6 alkyl or C _{3- 6} cycloalkyl, the alkyl or cycloalkyl is optionally substituted by the following substituents: F, Cl, C _1-6 alkyl or hydroxyl; R ² and R ³ are each independently selected from H, F, Cl or C _1-6 alkyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H or C _1-6 alkyl; R ⁸ , R ⁹ are each independently selected from H, F, Cl, C _1-6 Alkyl, which is optionally substituted by the following substituents: cyano, F or Cl, or R ⁸ and R ⁹ together form C _2-6 alkenyl or C _3-6 cycloalkyl; R ¹⁰ is selected from H. The tetrahydroindazole compound and its stereoisomers, pharmaceutically acceptable salts, hydrates or N-oxides according to claim 4, wherein: R ¹ is selected from

,

,

or

R ² , R ³ are each independently selected from H, F or methyl; R ⁴ , R ⁵ , R ⁶ , R ⁷ are each independently selected from H; R ⁸ , R ⁹ are each independently selected from H, F, isopropyl , or R ⁸ and R ⁹ together form 2-methylpropenyl or cyclopropyl; R ¹⁰ is selected from H; R ¹¹ is selected from

. The tetrahydroindazole compound and its stereoisomer, pharmaceutically acceptable salt, hydrate or N-oxide according to claim 1, wherein the compound structure is as follows:

The tetrahydroindazole compound and its stereoisomer, pharmaceutically acceptable salt, hydrate or N-oxide according to claim 6, wherein the compound structure is as follows:

A pharmaceutical composition, the composition comprising: an effective dose of the tetrahydroindazole compound represented by the general formula (I) according to any one of claim items 1 to 7 and a stereoisomer thereof, pharmaceutically acceptable The accepted salt, hydrate or N-oxide, or further comprises one or more other therapeutic agents and a pharmaceutically acceptable carrier or excipient. A tetrahydroindazole compound represented by general formula (I) as described in any one of claims 1 to 7 and its stereoisomers, pharmaceutically acceptable salts, hydrates or N-oxides or claims 8. Use of the composition in the preparation of a medicament for treating diseases mediated by CB2 receptors. The use according to claim 9, wherein the disease mediated by CB2 receptors is pain. The use according to claim 10, wherein the pain is selected from the group consisting of: bone pain, arthralgia, muscle pain, dental pain, migraine, headache, inflammatory pain, neuropathic pain, abdominal pain associated with Crohn's disease, adverse effects due to therapeutic agents Pain due to action and pain associated with a disease selected from the group consisting of osteoarthritis, cancer, multiple sclerosis, allergic response, nephritic syndrome, scleroderma, thyroiditis, diabetic neuropathy, fibromyalgia , HIV-related neuropathy, sciatica, and autoimmune diseases. The use according to claim 11, wherein the pain is selected from neuropathic pain, abdominal pain associated with Crohn's disease.