Heterocyclic fused ring compounds, pharmaceutical compositions thereof, and uses thereof

Abstract

The present invention belongs to the field of pharmaceuticals and relates to a class of heterocyclic fused ring compounds, a pharmaceutical composition thereof, and a use thereof. Specifically, the present invention relates to a compound represented by formula (I), or a stereoisomer, tautomer, nitric oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of the compound represented by formula (I). The compounds and a pharmaceutical composition thereof of the present invention, as PKR agonists, can be used for the preparation of a medicament for preventing or treating PKR-related diseases, and in particular, can be used for the preparation of a medicament for treating anemia.

Description

Heterocyclic ring compounds, pharmaceutical compositions thereof and uses thereof TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, in particular, the present application relates to a new class of heterocyclic ring compounds as PKR agonists, pharmaceutical compositions thereof, and the use of the compounds and pharmaceutical compositions thereof in the preparation of drugs for the treatment of PKR related diseases. BACKGROUND

[0002] Pyruvate kinase (PK, EC 2.7.1.40) catalyzes the conversion of phosphoenolpyruvate to pyruvate in the glycolytic pathway, red blood cells lack mitochondria, and glycolysis is the only way to provide energy, PK deficiency causes damage leading to red blood cell destruction, which is the most common cause of hereditary hemolytic anemia. Human PK includes four isozymes (L, R, M1, M2), PKR in mature red blood cells, PKM1 in brain and heart; PKL in kidney and liver and PKM2 in intestinal cells. PKR is the only isozyme found in normal mature red blood cells; PKM2 exists in erythroid precursor cells, and as the precursor cells differentiate, the enzyme in the cell changes from PKM2 to PKR (precursor red blood cells mainly PKM2, mature red blood cells mainly PKR).

[0003] Thalassemias (also known as dysglobulinemia) is a group of recessive genetic diseases characterized by reduced or no hemoglobin production and varying degrees of chronic anemia. According to the classification and clinical characteristics of the International Thalassemia Federation, thalassemia can be divided into transfusion-dependent thalassemia (TDT) and non-transfusion-dependent thalassemia (NTDT), and patients with transfusion-dependent thalassemia need to rely on blood transfusion for life to maintain life, and if not transfused, will have serious complications and early death. Thalassemia is due to mutations in genes encoding alpha and / or beta globin, imbalance in globin chain production, leading to excessive precipitation of alpha or beta globin, ineffective erythropoiesis and hemolysis; the removal of excess globin chains increases the demand for adenosine triphosphate (ATP) driven mechanisms, and efficient ATP production is essential for maintaining red blood cell function and membrane integrity, and ATP supply is often insufficient to meet the increased metabolic needs of thalassemia red blood cells, leading to premature cell death; glycolysis is the main pathway for red blood cells to produce energy, and PKR is a key enzyme for regulating ATP production through glycolysis, and increasing ATP synthesis through PKR activation may improve thalassemia red blood cell adaptability and survival, thereby reducing anemia and other clinical sequelae.

[0004] The research literature and patents of small molecule drugs targeting PKR have been disclosed, wherein FORMA Therapeutics Inc. discloses a pyrrolopyrrole compound as a PKR agonist in WO2018175474 and WO2020061255. Global Blood Therapeutics Inc. discloses a pyrrolidinopyrazole compound as a pyruvate kinase activator in WO2021202796. Agios Pharmaceuticals Inc. discloses azetidine aryl sulfonamide derivative compounds as pyruvate kinase modulators in WO2012083246, WO2011002817, WO2012092442, WO2014139325 and the like.

[0005] There are few pyruvate kinase agonists in clinical development at present, wherein Mitapivat developed by Agios Therapeutics for treating thalassemia is in clinical phase III research stage; Etavopivat developed by Forma Therapeutics is in clinical phase II research stage. Mitapivat has a short half-life, and the clinical use of twice-a-day administration greatly reduces the patient compliance; although Etavopivat is administered once a day, the high clinical administration dose is caused by poor in-vitro activity. Based on the prospect of PKR agonists in treating anemia and other diseases, it is urgent to develop a new type of PKR agonist with better drugability and stronger efficacy. SUMMARY

[0006] The present application provides a compound, or a pharmaceutical composition thereof, which can act as a PKR agonist. The present application also relates to the use of the compound or the pharmaceutical composition thereof for the preparation of a medicament for the treatment of a disease and / or a condition by activating the activity of PKR, in particular thalassemia.

[0007] In one aspect, the present application provides a compound represented by formula (I), or a stereoisomer, a tautomer, a nitroso, a hydrate, a solvate, a metabolite, a pharmaceutically acceptable salt or a prodrug of the compound represented by formula (I),

[0008] wherein,

[0009] R 1 is C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6hydroxyalkyl, C 1-6 alkoxy C 1-6 alkyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, C 6-10 aryl or 5-12 membered heteroaryl, wherein the C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 1-6 alkoxy C 1-6 alkyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, C 6-10 aryl and 5-12 membered heteroaryl are each independently optionally substituted with 1, 2, 3, 4, or 5 substituents selected from the group consisting of D, -OH, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 deuteroalkoxy, C 1-6 haloalkoxy, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl, 5-12 membered heteroaryl, -OR a , -C(=O)R a , -C(=O)OR a , -NR b R c , -C(=O)NR b R c , -NR b C(=O)R c , -S(O)R a , -S(O)2R a , -S(O)2NR b R c , and -NR b S(O)2R c ;

[0010] R 2 is H, D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 1-6 alkoxyC 1-6 alkyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, C 6-10 aryl or 5-12 membered heteroaryl, wherein the C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 1-6 alkoxyC 1-6 alkyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, C 6-10 aryl and 5-12 membered heteroaryl are each independently optionally substituted with 1, 2, 3, 4, or 5 substituents selected from the group consisting of D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxyC 1-6 alkoxy, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, -OR d , -C(=O)R d , -C(=O)OR d , -NR e R f , -C(=O)NR e R f , -NR e C(=O)R f , -S(O)R d , -S(O)2R d , -S(O)2NR e R f , and -NR e S(O)2R f ;

[0011] R 3 and R 4 are each independently H, D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 1-6 alkylamino, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl or 3-8 membered heterocyclyl, wherein said C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 1-6 alkylamino, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl and 3-8 membered heterocyclyl are each independently optionally substituted with 1, 2, 3, or 4 substituents selected from the group consisting of D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, oxo, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl and 3-6 membered heterocyclyl;

[0012] or, R 3 , R 4 and the carbon atom to which they are attached together form a C 3-8 cycloalkyl or 3-8 membered heterocyclyl, wherein said C 3-8 cycloalkyl and 3-8 membered heterocyclyl are each independently optionally substituted with 1, 2, or 3 substituents selected from the group consisting of D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, oxo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy and C 1-6 haloalkoxy;

[0013] R a , R b , R c , R d , R e and R f are each independently H, D, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10aryl or 5-12 membered heteroaryl, wherein the C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl and 5-12 membered heteroaryl are each independently optionally substituted with 1, 2, 3, or 4 substituents selected from the group consisting of D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy and C 1-6 haloalkoxy.

[0014] In some embodiments, R 1 is C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 haloalkyl, C 1-4 aminoalkyl, C 1-4 cyanoalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxy C 1-4 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl or 5-10 membered heteroaryl, wherein the C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 haloalkyl, C 1-4 aminoalkyl, C 1-4 cyanoalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxy C 1-4 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl and 5-10 membered heteroaryl are each independently optionally substituted with 1, 2, 3, 4, or 5 substituents selected from the group consisting of D, -OH, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 deuteroalkoxy, C 1-4 haloalkoxy, C 3-6 cycloalkyl, 3-6 membered heterocyclyl, C 6-10 aryl, 5-10 membered heteroaryl, -OR a , -C(=O)R a , -C(=O)OR a , -NRb R c , -C(=O)NR b R c , -NR b C(=O)R c , -S(O)R a , -S(O)2R a , -S(O)2NR b R c and -NR b S(O)2R c substituted.

[0015] In some embodiments, R 1 is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, ethenyl, ethynyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylen, ethoxymethylen, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, wherein said methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, ethenyl, ethynyl, -CH2F, -CHF2, -CH2CF2, -CH2CF3, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylen, ethoxymethylen, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, each independently optionally substituted with 1, 2, 3, 4, or 5 substituents selected from D, -OH, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, i-propoxy, deuterated methoxy, deuterated ethoxy, difluoromethoxy, trifluoromethoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, oxopyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, indenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, -OR a a a b c b c b c a a b c b c each independently optionally substituted with 1, 2, 3, 4, or 5 substituents selected from D, -OH, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, i-propoxy, deuterated methoxy, deuterated ethoxy, difluoromethoxy, trifluoromethoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, oxopyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, indenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, -OR

[0016] In some embodiments, R 2 is H, D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 aminoalkyl, C 1-4 cyanoalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxyC 1-4 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl, or 5-10 membered heteroaryl, wherein said C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 haloalkyl, C 1-4 alkoxy, C​​​​​​​​​​​​​​1-4 Halogenated alkoxy groups, C 1-4 aminoalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy C 1-4 Alkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic groups, C 6-10 The aryl group and the 5-10 heteroaryl group are each independently and optionally surrounded by 1, 2, 3, 4 or 5 groups selected from D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, hydroxyl C 1-4 Alkoxy, C 3-6 cycloalkyl, 3-6 membered heterocyclic groups, -OR d -C(=O)R d -C(=O)OR d -NR e R f -C(=O)NR e R f -NR e C(=O)R f -S(O)R d -S(O)2R d -S(O)2NR e R f and -NR e S(O)2R f The substituents are replaced by the substituents.

[0017] In some implementation schemes, R 2 The following are not part of the given sequence: H, D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, vinyl, ethynyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, isopropoxy, difluoromethoxy, trifluoromethoxy, aminomethyl, aminoethyl, cyanomethyl Cyclopropyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl, azacyclobutyl, pyrrolyl, tetrahydrofuranyl, piperidinyl, piperazine, morpholinyl, phenyl, indole, naphthyl, pyrrolyl, pyrazolyl, imidazoleyl, triazolyl, tetraazolyl, furanyl, thiophenyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyridazinyl The terms methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, vinyl, ethynyl, -CH2F, -CHF2, -CH2CF2, -CH2CF3, methoxy, ethoxy, isopropoxy, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl, azacyclobutyl, pyrrolyl, tetrahydrofuranyl, piperidinyl, piperazine, morpholinyl, phenyl, indole, naphthyl, pyrrolyl, pyrazolyl, imidazoleyl, triazolyl, tetraazolyl, furanyl, thiophenyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyrazinyl Each of the following is independently and optionally selected from 1, 2, 3, 4, or 5 groups: D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, isopropoxy, difluoromethoxy, trifluoromethoxy, hydroxymethoxy, hydroxyethoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetyl, aziroxy, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, -OR d -C(=O)R d -C(=O)OR d -NR e R f -C(=O)NR e R f -NR e C(=O)R f -S(O)R d -S(O)2R d -S(O)2NR e R f and -NR e S(O)2R f The substituents are replaced by the substituents.

[0018] In some implementation schemes, R 3 and R 4 Each of the following can be independently identified as H, D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy groups, C 1-4 Alkylamino, C1-4 aminoalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 3-6 Cycloalkyl or 3-6 membered heterocyclic group, wherein the C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy groups, C 1-4 Alkylamino, C 1-4 aminoalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 3-6 The cycloalkyl group and the 3-6 membered heterocyclic group are each independently and optionally surrounded by 1, 2, 3 or 4 groups selected from D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, oxo, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy groups, C 3-6 Substituents of cycloalkyl and 3-6 membered heterocyclic groups;

[0019] Or, R 3 R 4 Together with the carbon atoms attached to them, they form C 3-6 Cycloalkyl or heterocyclic group consisting of 3-6 atoms, wherein the C 3-6 The cycloalkyl group and the heterocyclic group consisting of 3-6 atoms are each independently and optionally surrounded by 1, 2 or 3 atoms selected from D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, oxo, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy and C 1-4 The substituents of the haloalkoxy group are replaced.

[0020] In some implementation schemes, R 3 and R 4each independently H, D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, vinyl, ethynyl, methoxy, ethoxy, i-propoxy, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, difluoromethoxy, trifluoromethoxy, N-methylamino, N-ethylamino, N,N-dimethylamino, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, or morpholinyl, wherein each of said methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, vinyl, ethynyl, methoxy, ethoxy, i-propoxy, -CH2F, -CHF2, -CH2CF2, -CH2CF3, N-methylamino, N-ethylamino, N,N-dimethylamino, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, and morpholinyl is independently optionally substituted with 1, 2, 3, or 4 substituents selected from D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, oxo, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, methoxy, ethoxy, i-propoxy, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, difluoromethoxy, trifluoromethoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, and morpholinyl;

[0021] or, R 3 , R 4 and the carbon atom to which they are attached together form a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, or morpholinyl, wherein each of said cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, and morpholinyl is independently optionally substituted with 1, 2, or 3 substituents selected from D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, oxo, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, i-propoxy, difluoromethoxy, and trifluoromethoxy.

[0022] In some implementation schemes, R a R b R c R d R e and R f Each is independently H, D, C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 cycloalkyl, 3-6 membered heterocyclic, C 6-10 aryl or 5-10 heteroaryl, wherein the C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 cycloalkyl, 3-6 membered heterocyclic, C 6-10 The aryl group and the 5-10 heteroaryl group are each independently and optionally surrounded by 1, 2, 3 or 4 groups selected from D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy and C 1-4 The substituents of the haloalkoxy group are replaced.

[0023] In some implementation schemes, R a R b R c R d R e and R feach independently H, D, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, indenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, or pyridazinyl, wherein each of said methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CH2CF2, -CH2CF3, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, indenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, and pyridazinyl is independently optionally substituted with 1, 2, 3, or 4 substituents selected from the group consisting of D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, i-propoxy, difluoromethoxy, and trifluoromethoxy.

[0024] In some embodiments, the compound of the present application is a compound of Formula (I-a) or (I-b), or a stereoisomer, a tautomer, a N-oxide, a hydrate, a solvate, a metabolite, a prodrug, a pharmaceutically acceptable salt or a prodrug of a compound of Formula (I-a) or (I-b),

[0025] wherein R 1 , R 2 , R 3 and R 4 each have the definitions as described herein.

[0026] In another aspect, the present application provides a pharmaceutical composition comprising a compound of the present application.

[0027] In some embodiments, the pharmaceutical composition of the present application further comprises a pharmaceutically acceptable adjuvant.

[0028] In some embodiments, the adjuvant of the present application includes, but is not limited to, a carrier, an excipient, a diluent, a vehicle, or a combination thereof. In some embodiments, the pharmaceutical composition can be in a liquid, solid, semi-solid, gel or spray form.

[0029] In another aspect, the present application provides use of the pharmaceutical composition described herein for the preparation of a medicament for preventing, treating or alleviating a PKR-associated disease.

[0030] In some embodiments, the PKR-associated disease described herein is thalassemia, pyruvate deficiency anemia, anemia associated with myelodysplastic syndrome (MDS), sickle cell anemia, megaloblastic anemia, aplastic anemia, iron deficiency anemia, hemolytic anemia, sickle cell disease, hereditary non-spherocytic hemolytic anemia, hereditary spherocytosis, hereditary elliptocytosis, abetalipoproteinemia, paroxysmal nocturnal hemoglobinuria, acquired hemolysis, tumor-related anemia, or chronic disease-related anemia.

[0031] In another aspect, the present application also provides a method for preventing or treating a PKR-associated disease, the method comprising administering to a patient a therapeutically effective amount of a compound described herein or a pharmaceutical composition thereof.

[0032] In another aspect, the present application relates to methods for preparing, isolating and purifying compounds of formula (I), (I-a) or (I-b).

[0033] Unless otherwise indicated, all stereoisomers, tautomers, nitroso isomers, hydrates, solvates, metabolites, pharmaceutically acceptable salts, and prodrugs of the compounds of the present application are within the scope of the application.

[0034] The term "pharmaceutically acceptable" includes that the substance or composition must be suitable chemical or toxicological, with other components of the formulation and for the mammal being treated.

[0035] Salts of the compounds of the present application also include salts of intermediates used in preparing or purifying compounds of formula (I), (I-a) or (I-b) or isolated enantiomers of compounds of formula (I), (I-a) or (I-b), but are not necessarily pharmaceutically acceptable salts.

[0036] The foregoing outlines some aspects of the present application, but is not limited to these aspects. Additional aspects will be set forth in more detail in the description that follows.

[0037] Detailed description of the application

[0038] Definitions and general terminology

[0039] Certain embodiments of the application are now described in detail by referring principally to and illustrating the described embodiments with the attached figures and chemical structures. The application intends to encompass all alternatives, modifications and equivalents that can be included within the scope of the application as defined by the claims. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many methods and materials similar or equivalent to those described herein. The application is not intended to be limited to the methods and materials described but instead includes any alternatives, modifications and equivalents falling within the scope of the claims. In the event that one or more of the attached claims is determined to be invalid by a court or other authoritative body, the inventor(s) hereby state that they will endeavor for the life of the patent(s) to endeavor to amend additional claims to substitute for any claims that are invalid to the extent of such invalidity, and to surrender a term of the patent life corresponding to the period that is deemed invalid.

[0040] It should further be recognized that certain of the features of the application, described in detail herein, are described in the context of a number of separate embodiments, but can also be provided in combination in a single embodiment. Conversely, various features of the application, described in the context of a single embodiment, can also be provided separately or in any appropriate subcombination.

[0041] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as those commonly understood by one of ordinary skill in the art to which this application belongs. All patents and publications referred to in this application are incorporated herein by reference in their entirety.

[0042] The term "subject" as used herein refers to an animal. Typically the animal is a mammal. Subject also refers to, for example, primates (e.g., humans, male or female), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds, and the like. In certain embodiments, the subject is a primate. In other embodiments, the subject is a human.

[0043] The term "patient" as used herein refers to a human (including adults and children) or other animal. In some embodiments, "patient" refers to a human.

[0044] The term "comprising" is used in the inclusive sense of "including" and not the exclusive sense (that is, "consisting of").

[0045] "Stereoisomers" refer to compounds which have the same chemical constitution, but differ in the arrangement of atoms or groups in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotamers), geometric isomers (cis / trans isomers), atropisomers, and the like. Unless otherwise specified, all stereoisomers and mixtures thereof of the structures depicted are intended to be covered herein by the description of the structure. In addition, unless otherwise specified, the structures depicted contain one or more double bonds, which are enantiomeric, diastereomeric, geometric isomers, or mixtures thereof.

[0046] The stereochemical definitions and rules generally follow S. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994.

[0047] Any mixture of stereoisomers resulting can be separated into the individual isomers by conventional separation techniques, or any mixture of stereoisomers can be converted into pure isomers, for example, by chromatographic and / or fractional crystallization techniques.

[0048] The term "tautomer" or "tautomerism" refers to structural isomers that exist in equilibrium with one another through a low energy barrier. If tautomerism is possible (as in solution), a chemical equilibrium of the tautomers can be reached. For example, protontautomer (also known as prototropic tautomer) includes interconversion by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomer includes interconversion by reorganization of some bonding electrons. A specific example of keto-enol tautomerism is the interconversion of pentane-2,4-dione and 4-hydroxypent-3-en-2-one tautomers. Another example of tautomerism is phenol-keto tautomerism. A specific example of phenol-keto tautomerism is the interconversion of pyridin-4-ol and pyridin-4(lH)-one tautomers. Unless otherwise indicated, all tautomeric forms of the compounds of the application are within the scope of the application.

[0049] As described herein, the compounds of the application can be independently optionally substituted with one or more substituents, such as the substituents of the general formulae above, or as in the specific examples, subgeneric classes, and generic classes of compounds embraced by the application. It will be appreciated that the terms "independently optionally substituted" or "optionally substituted" are used interchangeably with the term "substituted or unsubstituted." In general, the term "substituted" means that one or more hydrogens of the designated moiety are replaced by a non-hydrogen moiety. Unless otherwise indicated, an optionally substituted moiety can be substituted at each substitutable position of the moiety. When a structure is presented wherein more than one position of the structure can be substituted with one or more substituents selected from a particular group, the substituents can be the same or different at each occurrence.

[0050] Also, it is to be understood that the description and examples in this application use the description "each occurrence" and "independently" interchangeably, and that these terms are to be interpreted in their broadest context unless otherwise specified.

[0051] Throughout various portions of the specification, the substituents of the compounds disclosed herein are disclosed by group or range. It is specifically intended that the application include each and every independent subcombination of the members of the groups and ranges. For example, the term "C 1-6 "alkyl" specifically refers to the individually disclosed methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl groups.

[0052] Throughout various portions of the specification, linking substituents are described. When the structure clearly requires a linking group, the Markush group recited for that group is to be construed as the linking group. For example, if the structure requires a linking group and the Markush group recited for that group recites "alkyl" or "aryl," then it is to be understood that the "alkyl" or "aryl" represents the linking alkylene or arylene group, respectively.

[0053] The term "alkyl" denotes a straight or branched chain monovalent hydrocarbon group containing from 1 to 20 carbon atoms, wherein the alkyl group can be optionally substituted with one or more substituents as described herein. In one embodiment, the alkyl group contains 1 to 6 carbon atoms, denoted as C 1-6 alkyl; in yet another embodiment, the alkyl group contains 1 to 4 carbon atoms, denoted as C 1-4 alkyl; in yet another embodiment, the alkyl group contains 1 to 4 carbon atoms, denoted as C 1-3Alkyl. Examples of alkyl groups include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), n-propyl (n-Pr, -CH2CH2CH3), i-propyl (i-Pr, -CH(CH3)2), n-butyl (n-Bu, -CH2CH2CH2CH3), i-butyl (i-Bu, -CH2CH(CH3)2), s-butyl (s-Bu, -CH(CH3)CH2CH3), t-butyl (t-Bu, -C(CH3)3), n-pentyl (-CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-l-butyl (-CH2CH2CH(CH3)2), 2-methyl-l-butyl (-CH2CH(CH3)CH2CH3), n-hexyl (-CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3), n-heptyl, n-octyl, and the like.

[0054] The term "alkylene" denotes a saturated divalent hydrocarbyl radical resulting from the removal of two hydrogen atoms from a saturated straight chain or branched chain hydrocarbon. In some embodiments, the alkylene group contains 1 to 6 carbon atoms, denoted as C 1-6 alkylene; in other embodiments, the alkylene group contains 1 to 4 carbon atoms, denoted as C 1-4 alkylene; in other embodiments, the alkylene group contains 1 to 3 carbon atoms, denoted as C 1-3 alkylene; in other embodiments, the alkylene group contains 1 to 2 carbon atoms, denoted as C 1-2 alkylene. Examples of alkylene groups include, but are not limited to: -CH2-, -CH2CH2-, -CH(CH3)CH2-, and the like.

[0055] The term "alkenyl" refers to a straight-chain or branched monovalent hydrocarbon group containing 2-12 carbon atoms, with at least one unsaturated site, i.e., one carbon-carbon sp. 2 The double bond, wherein the alkenyl group may optionally be substituted by one or more substituents described in this invention, including the orientation of "cis" and "trans", or the orientation of "E" and "Z". In one embodiment, the alkenyl group comprises 2-6 carbon atoms, denoted as C 2-6 Alkenyl group; in yet another embodiment, the alkenyl group comprises 2-4 carbon atoms, denoted as C1 2-4 Alkenyl groups. Examples of alkenyl groups include, but are not limited to, vinyl (-CH=CH2), allyl (-CH2CH=CH2), 1-propenyl (i.e., propenyl, -CH=CH-CH3), etc.

[0056] The term "alkynyl" refers to a straight-chain or branched monovalent hydrocarbon group containing 2-12 carbon atoms, wherein there is at least one unsaturated site, i.e., one carbon-carbon sp triple bond, wherein the alkynyl group may optionally be substituted by one or more substituents described in this invention. In one embodiment, the alkynyl group comprises 2-6 carbon atoms, denoted as C1. 2-6 Alkynyl group; in yet another embodiment, the alkynyl group comprises 2-4 carbon atoms, denoted as C0. 2-4 Alkynyl. Examples of alkynyl groups include, but are not limited to, ethynyl (-C≡CH), propynyl (-CH2C≡CH), 1-propynyl (-C≡C-CH3), etc.

[0057] The term "cyanoalkyl" refers to an alkyl group substituted with one or more cyano groups, wherein the cyano and alkyl groups have the definitions described herein. In some embodiments, "cyanoalkyl" refers to an alkyl group substituted with one cyano group. In some embodiments, "cyanoalkyl" is C10. 1-6 Cyanoalkyl, i.e., C64 substituted with one or more cyano groups. 1-6 Alkyl group. In some preferred embodiments, C 1-6 Cyanoalkyl is a C group substituted with one cyano group. 1-6 Alkyl. In other embodiments, "cyanoalkyl" is C1 1-4 Cyanoalkyl, i.e., C4 groups substituted with one or more cyano groups. 1-4 Alkyl groups. Examples of cyanoalkyl groups include, but are not limited to, -CH2CN, -CH2CH2CH2CH2CN, -CH2CH2CN, -CH2CH(CN)CH2CH2CN, -CH2CH(CN)CH2CH(CH3)CN, etc.

[0058] The term "aminoalkyl" denotes an alkyl group substituted with one or more amino groups, wherein the amino and alkyl groups have the definitions as set forth herein. In some embodiments, "aminoalkyl" denotes an alkyl group substituted with one amino group. In some preferred embodiments, the C 1-6 aminoalkyl group is a C 1-6 alkyl group substituted with one or more amino groups. In some preferred embodiments, the C 1-6 aminoalkyl group is a C 1-6 alkyl group substituted with one amino group. In some embodiments, "aminoalkyl" denotes a C 1-4 aminoalkyl group is a C 1-4 alkyl group substituted with one or more amino groups. In some preferred embodiments, the C

[0059] The term "hydroxyalkyl" denotes an alkyl group substituted with one or more hydroxyl groups, wherein the alkyl and hydroxyl groups have the definitions as set forth herein. In some embodiments, hydroxyalkyl denotes an alkyl group substituted with 1, 2, 3, or 4 hydroxyl groups. In some embodiments, hydroxyalkyl denotes an alkyl group substituted with one or two hydroxyl groups. In some embodiments, hydroxyalkyl denotes a C 1-6 hydroxyalkyl group is a C 1-6 alkyl group substituted with one or more hydroxyl groups, preferably a C 1-6 hydroxyalkyl group denotes a C 1-6 alkyl group substituted with one hydroxyl group. In some embodiments, hydroxyalkyl denotes a C 1-4 hydroxyalkyl group. In some embodiments, hydroxyalkyl denotes a C 1-3 hydroxyalkyl group. Examples of hydroxyalkyl groups include, but are not limited to, -CH2OH, -CH2CH2OH, -CH2CH2CH2OH, -CH2CH2CH2CH2OH, -CH2CH(OH)CH2CH2OH, -CH2CH(OH)CH2CH(CH3)OH, and the like.

[0060] The term "hydroxyalkoxy" denotes an alkoxy group substituted with one or more hydroxyl groups, wherein the alkoxy and hydroxyl groups have the definitions as set forth herein. In some embodiments, hydroxyalkoxy denotes an alkoxy group substituted with 1, 2, 3, or 4 hydroxyl groups. In some embodiments, hydroxyalkyl denotes an alkoxy group substituted with one or two hydroxyl groups. In some embodiments, hydroxyalkoxy denotes a hydroxyl 1-6 alkoxy group, i.e., a C 1-6 alkoxy group substituted with one or more hydroxyl groups, preferably a hydroxyl1-6 Alkoxy represents C 1-6 Alkoxy represents C Examples of hydroxyalkyl include, but are not limited to, -OCH2OH (hydroxymethoxy), -OCH2CH2OH (hydroxyethoxy), -OCH2CH2CH2OH (hydroxypropoxy), and the like.

[0061] The term "haloalkyl" represents an alkyl group substituted with one or more halogen atoms, wherein alkyl and halogen have the meaning as described herein. In some embodiments, haloalkyl is C 1-6 Haloalkyl represents C 1-6 Haloalkyl represents C 1-4 Haloalkyl represents C 1-4 Haloalkyl represents C 1-3 Haloalkyl represents C 1-3 Haloalkyl represents C Examples include, but are not limited to, monofluoromethyl, difluoromethyl, trifluoromethyl, 1-fluoroethyl, 2-fluoroethyl, 1,2-difluoroethyl, 1,1-difluoroethyl, 2,2-difluoroethyl, monochloromethyl, dichloromethyl, trichloromethyl, 2-chloroethyl, 1-chloroethyl, 1,2-dichloroethyl, 1,1-dichloroethyl, 2,2-dichloroethyl, 1,1-dibromoethyl, and the like.

[0062] The term "alkoxy" represents an alkyl group attached to the remainder of the molecule through an oxygen atom, wherein alkyl group has the meaning as described herein. Unless otherwise specifically indicated, the alkoxy group contains 1 to 12 carbon atoms. In one embodiment, the alkoxy group contains 1 to 6 carbon atoms, representing C 1-6 Alkoxy represents C 1-4 Alkoxy represents C 1-3Alkoxy. The alkoxy group can optionally be substituted with one or more substituents described herein. Examples of alkoxy groups include, but are not limited to, methoxy (MeO, -OCH3), ethoxy (EtO, -OCH2CH3), 1-propoxy (n-PrO, n-propoxy, -OCH2CH2CH3), 2-propoxy (i-PrO, i-propoxy, -OCH(CH3)2), 1-butoxy (n-BuO, n-butoxy, -OCH2CH2CH2CH3), 2-methyl-l-propoxy (i-BuO, i-butoxy, -OCH2CH(CH3)2), 2-butoxy (s-BuO, s-butoxy, -OCH(CH3)CH2CH3), 2-methyl-2-propoxy (t-BuO, t-butoxy, -OC(CH3)3), 1-pentoxy (-OCH2CH2CH2CH2CH3), 2-pentoxy (-OCH(CH3)CH2CH2CH3), 3-pentoxy (-OCH(CH2CH3)2), 2-methyl-2-butoxy (-OC(CH3)2CH2CH3), 3-methyl-2-butoxy (-OCH(CH3)CH(CH3)2), 3-methyl-l-butoxy (-OCH2CH2CH(CH3)2), 2-methyl-l-butoxy (-OCH2CH(CH3)CH2CH3), and the like.

[0063] The term "haloalkoxy" denotes an alkoxy group substituted with one or more halogen, wherein alkoxy and halogen have the definitions as given herein. In some embodiments, haloalkoxy denotes haloalkoxy containing 1 to 6 carbon atoms, i.e. C 1-6 haloalkoxy; in other embodiments, haloalkoxy denotes haloalkoxy containing 1 to 4 carbon atoms, i.e. C 1-4 haloalkoxy; in other embodiments, haloalkoxy denotes haloalkoxy containing 1 to 3 carbon atoms, i.e. C 1-3 haloalkoxy. Examples of haloalkoxy include, but are not limited to, monofluoromethoxy (-OCH2F), difluoromethoxy (-OCF2), trifluoromethoxy (-OCF3), 2-fluoroethoxy (-OCH2CH2F), and the like.

[0064] The terms "alkylamino" and "alkylamino" are used interchangeably and include "N-alkylamino" and "N,N-dialkylamino", wherein the amino group is substituted with one or two alkyl substituents, respectively. In some embodiments, the alkylamino group is one or two C 1-6 alkyl groups attached to a nitrogen atom forming a lower alkylamino group. In other embodiments, the alkylamino group is one or two C 1-4lower alkylamino group formed by the attachment of an alkyl group to a nitrogen atom. In still other embodiments, alkylamino is one or two C 1-3 lower alkylamino group formed by the attachment of an alkyl group to a nitrogen atom. Suitable alkylamino groups can be monoalkylamino or dialkylamino, examples of alkylamino groups include, but are not limited to, N-methylamino, N-ethylamino, N,N-dimethylamino, N,N-diethylamino, and the like.

[0065] The term "cycloalkyl" denotes a monovalent saturated monocyclic or bicyclic carbon ring system of 3 to 12 carbon atoms, wherein a -CH2- group in the carbon ring can optionally be replaced by -C(=O)- (or -(CO)-). In one embodiment, cycloalkyl comprises 3 to 10 carbon atoms, i.e., C 3-10 cycloalkyl; in another embodiment, cycloalkyl comprises 3 to 7 carbon atoms, i.e., C 3-7 cycloalkyl; in another embodiment, cycloalkyl comprises 3 to 6 carbon atoms, i.e., C 3-6 cycloalkyl; in another embodiment, cycloalkyl comprises 3 to 5 carbon atoms, i.e., C 3-5 cycloalkyl; in another embodiment, cycloalkyl is a monocyclic cycloalkyl group comprising 3 to 7 carbon atoms, i.e., C 3-7 monocyclic cycloalkyl. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, octahydro-lH-indenyl, octahydro- pentalenyl, and the like. Examples of -CH2- groups in the carbon ring that can be replaced by -C(=O)- include, but are not limited to, cyclopentanone, cyclobutanone, and the like.

[0066] The term "heterocycle" or "heterocyclyl" denotes a saturated or partially unsaturated monocyclic, bicyclic or tricyclic ring system comprising 3-12 ring atoms, wherein at least one ring atom is selected from the group consisting of nitrogen, sulfur and oxygen atoms; wherein the heterocycle or heterocyclyl is non-aromatic and does not contain any aromatic rings. When a heterocycle is attached to the remainder of the molecule through a single bond, the heterocycle is designated as a monovalent heterocyclyl group. Unless indicated otherwise, the heterocycle group can be carbon-based or nitrogen-based and the -CH2- group can optionally be replaced by -C(=O)-. A sulfur atom of the ring can optionally be oxidized to the S-oxide or the S-oxide. A nitrogen atom of the ring can optionally be oxidized to the N-oxide. In some embodiments, the heterocycle or heterocyclyl group consists of 3-10 atoms, designated as 3-10 membered heterocycle or 3-10 membered heterocyclyl; in other embodiments, the heterocycle or heterocyclyl group consists of 3-9 atoms, designated as 3-9 membered heterocycle or 3-9 membered heterocyclyl; in other embodiments, the heterocycle or heterocyclyl group consists of 5-9 atoms, designated as 5-9 membered heterocycle or 5-9 membered heterocyclyl; in other embodiments, the heterocycle or heterocyclyl group consists of 3-6 atoms, designated as 3-6 membered heterocycle or 3-6 membered heterocyclyl; in other embodiments, the heterocycle or heterocyclyl group consists of 5-6 atoms, designated as 5-6 membered heterocycle or 5-6 membered heterocyclyl. In other embodiments, the heterocycle or heterocyclyl group consists of a monocyclic ring of 3-7 atoms, designated as 3-7 membered monocyclic heterocycle or 3-7 membered monocyclic heterocyclyl. Examples of the heterocycle include, but are not limited to, oxirane, aziridine, azetidine, oxetane, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, thiazolidine, pyrazolidine, pyrazoline, oxazolidine, imidazolidine, piperidine, piperazine, morpholine, 3,8-diazabicyclo[3.2.1]octane, 3,6-diazabicyclo[3.1.1]heptane, 2,5-diazabicyclo[2.2.2]octane. The heterocyclyl group includes, but is not limited to, oxiranyl, aziridinyl, azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, thiazolidinyl, pyrazolidinyl, pyrazolinyl, oxazolidinyl, imidazolidinyl, piperidinyl, piperazinyl, morpholinyl, and the like.

[0067] The term "aryl" denotes a monovalent, monocyclic, bicyclic and tricyclic carbon ring system containing 6-14 ring atoms, or 6-12 ring atoms, or 6-10 ring atoms, wherein at least one ring system is aromatic, wherein each ring system contains 3-7 atoms to form a ring. In some embodiments, the aryl group contains 6-12 ring atoms, designated as C 6-12 aryl or 6-12 membered aryl. In some embodiments, the aryl group contains 6-10 ring atoms, designated as C 6-10 aryl or 6-10 membered aryl. Examples of aryl groups can include phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl and anthracene.

[0068] The term "heteroaryl" or "heteroaromatic" refers to a monovalent monocyclic, bicyclic, or tricyclic ring system containing 5-14 ring atoms, or 5-12 ring atoms, or 5-10 ring atoms, or 5-6 ring atoms, at least one of which is aromatic, and at least one of which contains one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur. The heteroaryl group is generally, but not necessarily, attached to the parent molecule through an aromatic ring of the heteroaryl group. When a -CH2- group is present in a heteroaryl group, the -CH2- group can optionally be replaced with -C(=O)-. Unless otherwise indicated, the heteroaryl group can be attached to the remainder of the molecule (e.g., the main structure in a general formula) through any reasonable site (which can be C or N). The term "heteroaryl" can be used interchangeably with the term "heteroaromatic" or "heteroaromatic compound". In some embodiments, the heteroaryl is a heteroaryl containing 5-12 ring atoms, denoted as 5-12 membered heteroaryl; in other embodiments, the heteroaryl is a heteroaryl containing 5-10 ring atoms, denoted as 5-10 membered heteroaryl; in other embodiments, the heteroaryl is a heteroaryl containing 5-6 ring atoms, denoted as 5-6 membered heteroaryl. Examples of heteroaryl groups include, but are not limited to, furanyl, imidazolyl, isoxazolyl, oxazolyl, pyrrolyl, pyrazolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, thiophenyl, thiazolyl, triazolyl, tetrazolyl, benzofuranyl, benzothiazolyl, benzopyridyl, benzimidazolyl, benzopyrrolyl, benzopyrazolyl, benzopyrrolidinyl, 2,3-dihydrobenzofuranyl, benzo[2,3-b][l,4]dioxine, pyrido[2,3-b][l,4]dioxine, benzo[2,3-b][l,4]oxazine, pyrido[2,3-b][l,4]oxazine, and the like.

[0069] The term "halogen" or "halo" means F (fluorine), Cl (chlorine), Br (bromine), or I (iodine).

[0070] The term "oxo" means =O.

[0071] The term "cyano" means -CN or -C≡N.

[0072] The term "mercapto" means -SH.

[0073] The term "hydroxy" means -OH.

[0074] The term "carboxy" means -C(=O)OH.

[0075] The term "amino" means -NH2.

[0076] The term "consisting of" or "consisting" means "including and limited to." The term "j-k atoms" or "j-k membered" means that the cyclic group consists of j-k ring atoms, including carbon atoms and / or O, N, S, P, and the like heteroatoms; j and k are each independently any non-zero natural number, and k > j; "j-k" includes j, k, and any natural number in between. For example, "3-8 atoms" or "3-8 membered," "3-6 atoms" or "3-6 membered," "5-10 atoms" or "5-10 membered," or "5-6 atoms" or "5-6 membered," means that the cyclic group consists of 3-8 (i.e., 3, 4, 5, 6, 7, or 8), 3-6 (i.e., 3, 4, 5, or 6), 5-10 (i.e., 5, 6, 7, 8, 9, or 10), or 5-6 (i.e., 5 or 6) ring atoms, including carbon atoms and / or O, N, S, P, and the like heteroatoms.

[0077] The term "prodrug" as used herein refers to a compound that is converted into a compound of Formula (I), (I-a), or (I-b) in vivo. Such conversion is affected by the hydrolysis of the prodrug in the blood or by enzymatic conversion of the prodrug in the blood or tissues to the parent structure. The prodrug form of the compounds of the present application can be an ester, and in the present application the ester can be a benzoate, an aliphatic (C 1-24 ) ester, an acyloxymethyl ester, a carbonate, a carbamate, and an amino acid ester. For example, a compound of the present application containing a hydroxyl group can be acylated to provide a prodrug form of the compound. Other prodrug forms include phosphate esters, such as those formed by phosphorylation of a hydroxyl group on the parent.

[0078] "Metabolite" refers to a product produced through metabolism of a specified compound or salt thereof in vivo. Metabolites of a compound can be identified using techniques known in the art, and have activities generally inherent to the compound (for example, inhibiting the activity of a target enzyme). Such products can be identified using standard analytical techniques, such as those described herein. The present application includes metabolites of the compounds of the present application, including those produced upon administration of the compound to a mammal for a period of time sufficient to yield a metabolic product.

[0079] As used herein, "pharmaceutically acceptable salt" refers to organic and inorganic salts of the compounds of the application. Pharmaceutically acceptable salts are well known in the art, for examples, see S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66: 1-19. Pharmaceutically acceptable, non-toxic acid addition salts include those derived from inorganic acids, including, but not limited to, hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, sulfuric, hydroxamic, and the like, as well as salts derived from relatively nontoxic organic acids such as, but not limited to, citric, mallic, tartaric, lactic, benzoic, ascorbic, succinic, malonic, and the like. The present application also contemplates the quaternization of any basic nitrogen-containing groups of the compounds described herein. Water or oil-soluble or dispersable products can be obtained by quaternization. Pharmaceutically acceptable salts further include appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed by the addition of inorganic or organic acids to a basic nitrogen-containing group.

[0080] As used herein, "solvate" refers to an association or complex of one or more solvent molecules and a compound of the present application. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, ethanolamine, or mixtures thereof. The term "hydrate" refers to the complex that contains water as the solvent.

[0081] The term "hydrate" can be used when the solvent is water. In one embodiment, one molecule of a compound of the present application can be associated with one molecule of water, such as a monohydrate; in another embodiment, one molecule of a compound of the present application can be associated with more than one molecule of water, such as a dihydrate; in yet another embodiment, one molecule of a compound of the present application can be associated with less than one molecule of water, such as a hemihydrate. It is noted that the hydrates of the present application retain the biological effectiveness of the non-hydrated form of the compounds. The term "treatment" or "treating" any disease or disorder, as used herein, in some embodiments means ameliorating the disease or disorder (i.e., causing regression or halting or reducing the progression of the disease or at least one clinical symptom thereof). In other embodiments, "treatment" or "treating" means alleviating or improving at least one physical parameter, including parameters that can not be perceptible to the subject. In other embodiments, "treatment" or "treating" means modulating the disease or disorder physically (e.g., stabilizing a perceptible symptom), physiologically (e.g., stabilizing a physical parameter), or both. In other embodiments, "treatment" or "treating" means preventing or delaying the onset, development, or worsening of a disease or disorder.

[0082] The terms "prevent" or "preventing" mean a reduction in risk of acquiring a disease or disorder (i.e., causing at least one of the clinical symptoms of the disease not to develop in a subject that can be, or predisposed to be, exposed to the disease but does not yet experience or exhibit symptoms of the disease).

[0083] The term "therapeutically effective amount" means an amount of a compound that, when administered to a subject for treating a disease, is sufficient to effect such treatment for the disease. The "therapeutically effective amount" will vary depending on the compound, the disease and its severity, and the condition, age, body weight, sex, of the subject to be treated.

[0084] Unless otherwise stated, all isotopic variations, stereoisomers, tautomers, hydrates, solvates, metabolites, pharmaceutically acceptable salts, and prodrugs of the compounds of the present application are intended to be covered herein by the scope of the present application.

[0085] In the structures disclosed herein, when the stereochemistry of any particular chiral atom is not indicated, then all stereoisomers of the structure are intended to be covered herein and are included as compounds of the present application. When stereochemistry is indicated by a solid wedge or dashed wedge, then the stereoisomer is specifically intended and defined.

[0086] Nitrooxides of the compounds of the present application are also within the scope of the present application. Nitrooxides of the compounds of the present application can be prepared by oxidation of the corresponding nitrogen-containing base using a common oxidizing agent (e.g., hydrogen peroxide) at elevated temperature in the presence of an acid such as acetic acid, or by reaction with a peracid in a suitable solvent such as peracetic acid in dichloromethane, ethyl acetate or methyl acetate, or 3-chloroperoxybenzoic acid in chloroform or dichloromethane.

[0087] The compounds of Formula (I), (I-a), or (I-b) can exist in a salt form.

[0088] Any formula given herein is also intended to represent hydrates, solvates, and polymorphs of the compounds. Any formula given herein is also intended to represent isotopically enriched forms of the compounds. Isotopically enriched compounds have the structure depicted by the general formula given herein except that one or more atoms are replaced by an atom having the selected atomic mass or mass number. Exemplary isotopes include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, chlorine, and iodine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, 17 O, 18 O, 18 F, 31 P, 32 P, 35 S, 36 Cl, and 125 I.

[0089] Description of the Compounds of the Invention

[0090] The present application provides a compound, or a pharmaceutical composition thereof, which can act as an agonist of PKR.

[0091] The present application further relates to the use of the compound, or a pharmaceutical composition thereof, for the preparation of a medicament for the treatment of a disease and / or a condition by activating PKR activity by the compound.

[0092] The superior properties of the compounds of the present application, such as half-life, clearance, selectivity, bioavailability, chemical stability, metabolic stability, membrane permeability, solubility, etc., can result in a reduction of side effects, an enlargement of the therapeutic index, or an improvement of tolerability, etc.

[0093] In one aspect, the present application provides a compound represented by Formula (I), or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, a metabolite, a prodrug, or a pharmaceutically acceptable salt of the compound represented by Formula (I),

[0094] wherein R 1 , R 2 , R 3 , and R 4 each has the definition as described in the present application.

[0095] In some embodiments, R 1 is H, D, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 1-6 alkoxy C 1-6 alkyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, C 6-10 aryl, or 5-12 membered heteroaryl, wherein said C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 1-6 alkoxy C 1-6 alkyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, C 6-10The aryl group and the 5-12 heteroaryl group are each independently and optionally surrounded by 1, 2, 3, 4 or 5 groups selected from D, -OH, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Deuterated alkoxy, C 1-6 Halogenated alkoxy groups, C 3-8 Cycloalkyl, 3-8 membered heterocyclic groups, C 6-10 Aryl, 5-12 heteroaryl, -OR a -C(=O)R a -C(=O)OR a -NR b R c -C(=O)NR b R c -NR b C(=O)R c -S(O)R a -S(O)2R a -S(O)2NR b R c and -NR b S(O)2R c Substituents are replaced by R; a R b and R c Each has the definition as described in this invention.

[0096] In some implementation schemes, R 1 For H, D, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Haloalkyl, C 1-4 aminoalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy C 1-4 Alkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic groups, C 6-10 aryl or 5-10 heteroaryl, wherein the C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Haloalkyl, C 1-4 aminoalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy C 1-4 Alkyl, C3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl and 5-10 membered heteroaryl are each independently optionally substituted with 1, 2, 3, 4, or 5 substituents selected from the group consisting of D, -OH, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 deuteroalkoxy, C 1-4 haloalkoxy, C 3-6 cycloalkyl, 3-6 membered heterocyclyl, C 6-10 aryl, 5-10 membered heteroaryl, -OR a , -C(=O)R a , -C(=O)OR a , -NR b R c , -C(=O)NR b R c , -NR b C(=O)R c , -S(O)R a , -S(O)2R a , -S(O)2NR b R c and -NR b S(O)2R c ; R a , R b and R c each have the definition as described in the present application.

[0097] In other embodiments, R 1 is H, D, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, ethenyl, ethynyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, The terms methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, vinyl, ethynyl, -CH2F, -CHF2, -CH2CF2, -CH2CF3, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl, aziroxybutyl, pyrrolyl, tetrahydrofuranyl, piperidinyl, piperazine, morpholinyl, phenyl, naphthyl, pyrrolyl, pyrazolyl, imidazoleyl, triazolyl, tetraazolyl, furanyl, thiophenyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyrazinyl Each of the following can be independently and optionally selected from 1, 2, 3, 4, or 5 groups: D, -OH, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, isopropoxy, deuterated methyl Oxygen, deuterated ethoxy, difluoromethoxy, trifluoromethoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl, aziridine, pyrrolyl, oxopyrrolyl, tetrahydrofuranyl, piperidinyl, piperazine, morpholinyl, phenyl, indole, naphthyl, pyrrolyl, pyrazolyl, imidazoleyl, triazolyl, tetrazolyl, furanyl, thiophenyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, -OR a -C(=O)R a -C(=O)OR a -NR b R c -C(=O)NR b R c -NR b C(=O)R c -S(O)R a -S(O)2R a -S(O)2NR b R c and -NR b S(O)2R c Substituents are replaced by R; a R b and R c Each has the definition as described in this invention.

[0098] In some implementation schemes, R 2 is H, D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, C1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 1-6 alkoxyC 1-6 alkyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, C 6-10 aryl or 5-12 membered heteroaryl, wherein the C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 1-6 alkoxyC 1-6 alkyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, C 6-10 aryl and 5-12 membered heteroaryl are each independently optionally substituted with 1, 2, 3, 4, or 5 substituents selected from the group consisting of D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxyC 1-6 alkoxy, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, -OR d , -C(=O)R d , -C(=O)OR d , -NR e R f , -C(=O)NR e R f , -NR e C(=O)R f , -S(O)R d , -S(O)2R d , -S(O)2NR e R f , and -NR e S(O)2R f ; R d , R e , and Rf each having the definition as described in the present application.

[0099] In some embodiments, R 2 H, D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 aminoalkyl, C 1-4 cyanoalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxyC 1-4 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl or 5-10 membered heteroaryl, wherein said C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 aminoalkyl, C 1-4 cyanoalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxyC 1-4 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl and 5-10 membered heteroaryl are each independently optionally substituted with 1, 2, 3, 4, or 5 groups selected from D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, hydroxyC 1-4 alkoxy, C 3-6 cycloalkyl, 3-6 membered heterocyclyl, -OR d , -C(=O)R d , -C(=O)OR d , -NR e R f , -C(=O)NR e R f , -NR e C(=O)R f , -S(O)R d , -S(O)2R d-S(O)2NR e R f and -NR e S(O)2R f ; R d , R e , and R f each have the definitions as described herein.

[0100] In other embodiments, R 2 is H, D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, ethenyl, ethynyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, i-propoxy, difluoromethoxy, trifluoromethoxy, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, indenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, wherein said methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, ethenyl, ethynyl, -CH2F, -CHF2, -CH2CF2, -CH2CF3, methoxy, ethoxy, i-propoxy, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, indenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, each independently optionally substituted with 1, 2, 3, 4, or 5 substituents selected from the group consisting of D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, i-propoxy, difluoromethoxy, trifluoromethoxy, hydroxymethoxy, hydroxyethoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, -OR d , -C(=O)Rd -C(=O)OR d -NR e R f -C(=O)NR e R f -NR e C(=O)R f -S(O)R d -S(O)2R d -S(O)2NR e R f and -NR e S(O)2R f Substituents are replaced by R; d R e and R f Each has the definition as described in this invention.

[0101] In some implementation schemes, R 3 and R 4 Each of the following can be independently identified as H, D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy groups, C 1-6 Alkylamino, C 1-6 aminoalkyl, C 1-6 Cyanoalkyl, C 1-6 Hydroxyalkyl, C 3-8 Cycloalkyl or 3-8 membered heterocyclic group, wherein the C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy groups, C 1-6 Alkylamino, C 1-6 aminoalkyl, C 1-6 Cyanoalkyl, C 1-6 Hydroxyalkyl, C 3-8 The cycloalkyl group and the 3-8 membered heterocyclic group are each independently and optionally surrounded by 1, 2, 3 or 4 groups selected from D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, oxo, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy groups, C 3-6Substituents of cycloalkyl and 3-6 membered heterocyclic groups;

[0102] Or, R 3 R 4 Together with the carbon atoms attached to them, they form C 3-8 Cycloalkyl or heterocyclic group consisting of 3-8 atoms, wherein the C 3-8 The cycloalkyl group and the heterocyclic group consisting of 3-8 atoms are each independently and optionally surrounded by 1, 2 or 3 atoms selected from D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, oxo, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy and C 1-6 The substituents of the haloalkoxy group are replaced.

[0103] In some implementation schemes, R 3 and R 4 Each of the following can be independently identified as H, D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy groups, C 1-4 Alkylamino, C 1-4 aminoalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 3-6 Cycloalkyl or 3-6 membered heterocyclic group, wherein the C 1-4 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy groups, C 1-4 Alkylamino, C 1-4 aminoalkyl, C 1-4 Cyanoalkyl, C 1-4 Hydroxyalkyl, C 3-6 The cycloalkyl group and the 3-6 membered heterocyclic group are each independently and optionally surrounded by 1, 2, 3 or 4 groups selected from D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, oxo, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy groups, C 3-6 Substituents of cycloalkyl and 3-6 membered heterocyclic groups;

[0104] or R 3 or R 4 and the carbon atom to which they are attached together form a C 3-6 cycloalkyl or 3-6 membered heterocyclyl, wherein said C 3-6 cycloalkyl and 3-6 membered heterocyclyl are each independently optionally substituted with 1, 2, or 3 substituents selected from the group consisting of D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, oxo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, and C 1-4 haloalkoxy.

[0105] In other embodiments, R 3 and R 4 are each independently H, D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, vinyl, ethynyl, methoxy, ethoxy, i-propoxy, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, difluoromethoxy, trifluoromethoxy, N-methylamino, N-ethylamino, N,N-dimethylamino, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, or morpholinyl, wherein said methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, vinyl, ethynyl, methoxy, ethoxy, i-propoxy, -CH2F, -CHF2, -CH2CF2, -CH2CF3, N-methylamino, N-ethylamino, N,N-dimethylamino, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, and morpholinyl are each independently optionally substituted with 1, 2, 3, or 4 substituents selected from the group consisting of D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, oxo, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, methoxy, ethoxy, i-propoxy, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, difluoromethoxy, trifluoromethoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, and morpholinyl;

[0106] or R 3 , R 4 and the carbon atom to which they are attached together form a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, or morpholinyl, wherein each cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, and morpholinyl is independently optionally substituted with 1, 2, or 3 substituents selected from D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, oxo, methyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, i-propoxy, difluoromethoxy, and trifluoromethoxy.

[0107] In some embodiments, R a , R b , R c , R d , R e , and R f are each independently H, D, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl, or 5-12 membered heteroaryl, wherein each of said C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl, and 5-12 membered heteroaryl is independently optionally substituted with 1, 2, 3, or 4 substituents selected from D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, and C 1-6 haloalkoxy.

[0108] In some embodiments, R a , R b , R c , R d , R e , and R f are each independently H, D, C 1-4 alkyl, C 1-4 haloalkyl, C 3-6 cycloalkyl, 3-6 membered heterocyclyl, C 6-10 aryl, or 5-10 membered heteroaryl, wherein each of said C 1-4 alkyl, C1-4 Haloalkyl, C 3-6 cycloalkyl, 3-6 membered heterocyclic, C 6-10 The aryl group and the 5-10 heteroaryl group are each independently and optionally surrounded by 1, 2, 3 or 4 groups selected from D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy and C 1-4 The substituents of the haloalkoxy group are replaced.

[0109] In other implementations, R a R b R c R d R e and R f Each of the following is independently H, D, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl, aziridine, pyrrolyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, indole, naphthyl, pyrrolyl, pyrazolyl, imidazoleyl, triazolyl, tetrazolyl, furanyl, thiophenyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, or pyridazinyl, wherein the methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CH2F, -CHF2, -CH2CF2, -CH2CF3, cyclopropyl, cyclobutyl, cyclo... The pentyl, cyclohexyl, oxacyclobutyl, aziridine, pyrrolyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, indole, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetraazolyl, furanyl, thiophenyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, and pyridazinyl groups are each independently and optionally substituted by 1, 2, 3, or 4 substituents selected from D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, isopropoxy, difluoromethoxy, and trifluoromethoxy groups.

[0110] In some embodiments, the compound of the present invention is a compound of formula (Ia), or a stereoisomer, tautomer, nitride, hydrate, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of a compound of formula (Ia).

[0111] wherein R 1 , R 2 , R 3 and R 4 each has the definition as described in the present application.

[0112] In some embodiments, the compound described in the present application is a compound as shown in formula (I-b), or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, a metabolite, a prodrug, a pharmaceutically acceptable salt of the compound shown in (I-b),

[0113] wherein R 1 , R 2 , R 3 and R 4 each has the definition as described in the present application.

[0114] In some embodiments, the compound described in the present application has the structure shown in formula (I-b), or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate, a metabolite, a prodrug, a pharmaceutically acceptable salt of the compound shown in (I-b),

[0115] In another aspect, the present application provides a pharmaceutical composition comprising a compound described in the present application.

[0116] In some embodiments, the pharmaceutical composition described in the present application further comprises a pharmaceutically acceptable adjuvant.

[0117] In some embodiments, the adjuvant described in the present application includes, but is not limited to, a carrier, an excipient, a diluent, a vehicle, or a combination thereof. In some embodiments, the pharmaceutical composition can be in a liquid, a solid, a semi-solid, a gel or a spray form.

[0118] In another aspect, the present application provides use of a pharmaceutical composition described in the present application in the manufacture of a medicament for preventing, treating or alleviating a PKR-associated disease.

[0119] In some embodiments, the PKR-related disease is thalassemia, pyruvate deficiency anemia, anemia associated with myelodysplastic syndrome (MDS), sickle cell anemia, megaloblastic anemia, aplastic anemia, iron deficiency anemia, hemolytic anemia, sickle cell disease, hereditary non-spherocytic hemolytic anemia, hereditary spherocytosis, hereditary elliptocytosis, abetalipoproteinemia, paroxysmal nocturnal hemoglobinuria, acquired hemolysis, tumor-related anemia, or chronic disease-related anemia.

[0120] In another aspect, the present application also provides a method for preventing or treating a PKR-related disease, the method comprising administering to a patient a therapeutically effective amount of a compound of the present application or a pharmaceutical composition thereof.

[0121] In another aspect, the present application relates to a method for preparing, isolating and purifying a compound of Formula (I), (I-a) or (I-b).

[0122] Unless otherwise indicated, all stereoisomers, tautomers, nitroso isomers, hydrates, solvates, metabolites, pharmaceutically acceptable salts, and prodrugs of the compounds of the present application are within the scope of the application.

[0123] The term "pharmaceutically acceptable" includes that the substance or composition must be suitable chemically and toxicologically, with respect to the other components of the formulation and the mammal being treated therewith.

[0124] Salts of the compounds of the present application also include salts of intermediates used in making or purifying a compound of Formula (I), (I-a) or (I-b), or isolated enantiomers of a compound of Formula (I), (I-a) or (I-b), but are not necessarily pharmaceutically acceptable salts.

[0125] Pharmaceutical compositions, formulations, administration and uses of the compounds of the present application

[0126] The pharmaceutical compositions of the present application feature a compound of Formula (I), (I-a) or (I-b), a compound listed in the present application, or a compound of the Examples, and a pharmaceutically acceptable carrier. The amount of compound in the pharmaceutical compositions of the present application is effective to treat or alleviate a PKR-mediated disease in a patient.

[0127] The compounds of the present application exist in free form or in a suitable, pharmaceutically acceptable derivative. According to the present application, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable prodrugs, salts, esters, salts of esters, or any other adduct or derivative which upon administration to a patient is capable of providing (directly or indirectly) a compound described in other aspects of the present application, a metabolite or residue thereof.

[0128] As described herein, the pharmaceutically acceptable compositions of the present application further comprise a pharmaceutically acceptable carrier, which as used herein includes any and all solvents, diluents, or other liquid vehicles, suspending agents, or dispersing agents, surfactants, isotonic agents, thickening agents, emulsifying agents, preservatives, solid binders or lubricants, and the like, as suited to the particular dosage form desired. General considerations relating to various pharmaceutically acceptable carriers can also be found in Remington: The Science and Practice of Pharmacy, 21st edition, 2005, ed. D.B. Troy, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J.C. Boylan, 1988-1999, Marcel Dekker, New York, the contents of which are incorporated herein by reference. Except insofar as any conventional carrier is incompatible with the compounds of the application, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutically acceptable composition, its use is contemplated to be within the scope of this application.

[0129] Suitable excipients for the pharmaceutical compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene- block polymers, wool fat, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, phosphate buffer solutions, and other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, coloring agents, releasing agents, coating agents, sweetening, flavoring, and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator.

[0130] In making the pharmaceutical compositions of the present application, the active ingredients will generally be mixed with an excipient, diluted by an excipient or enclosed within such a carrier in the form of, e.g., a capsule, sachet, paper, or other container. If the excipient serves as a diluent, it can be a solid, semi-solid, or liquid material, which acts as a vehicle, carrier or medium for the active ingredient. Examples of suitable carriers include but are not limited to magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, gelatin, acacia, methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoa butter, and the like. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, and capsules, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solid forms or in liquid medium), ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders. In one embodiment, the composition is formulated for oral administration. In one embodiment, the composition is formulated as a tablet or capsule.

[0131] The compounds or pharmaceutical compositions of this application can be administered in a form of oral dosage such as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions. They can also be administered in intravenous (bolus injection or infusion), intraperitoneal, subcutaneous or intramuscular form, all using dosage forms well known in the pharmaceutical art. They can be administered alone, but generally will be administered in admixture with a pharmaceutical carrier selected with due regard to the intended route of administration and standard pharmaceutical practice.

[0132] The compounds or pharmaceutical compositions of this application can be administered in a form of oral dosage such as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions. They can also be administered in intravenous (bolus injection or infusion), intraperitoneal, subcutaneous or intramuscular form, all using dosage forms well known in the pharmaceutical art. They can be administered alone, but generally will be administered in admixture with a pharmaceutical carrier selected with due regard to the intended route of administration and standard pharmaceutical practice.

[0133] The compounds or pharmaceutical compositions of this application can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.

[0134] The compounds or pharmaceutical compositions of the present application can also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyethylene glycol, pyran co-polymers, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspartamide-phenol, or polyethylene oxide-polylysine substituted with palmitoyl residues. Furthermore, the compounds of the present application can be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyglycolic acid, copolymer of poly-lactic and poly-glycolic acid, polyepsilon caprolactone, polyhydroxybutyric acid, polyortho esters, polyacetals, polydihydropyrans, polycyanoacrylates, and cross-linked or amphipathic block co-polymers of hydrogels.

[0135] The dosage regimen for compounds or pharmaceutical compositions of the present application will vary according to various factors, such as the pharmacokinetic characteristics of the particular agent and its mode of administration; the species, age, sex, health, medical condition, and body weight of the recipient; the nature and extent of the symptoms; the kind of concurrent treatment(s); the frequency with which treatment is to be effected; the route of administration, the renal and hepatic function of the patient, and the effect desired. A physician or veterinarian will determine and prescribe the effective amount of the drug that is appropriate for a particular set of circumstances. It is expected that the dosage regimen will include 1 to 4 doses per day.

[0136] According to general principles, to achieve the desired effects, the dosage of each active ingredient used will range from about 0.001 to 1000 mg / kg body weight per day, orally, preferably from about 0.01 to 100 mg / kg body weight per day. The compounds of the present application can be administered once a day, or can be administered in two, three or four doses per day.

[0137] Each unit dose of the dosage form (pharmaceutical composition) suitable for administration can contain from about 1 mg to about 100 mg of the active ingredient. In these pharmaceutical compositions, the weight of active ingredient generally will comprise from about 0.5 to 95 percent of the total weight of the composition.

[0138] The compounds and compositions of the present application can be administered alone or in combination with other compounds or other therapeutic agents. The compounds or compositions of the present application can be administered simultaneously or sequentially with the other therapeutic agent(s), by the same or different routes of administration. The compounds of the present application can be included in a single formulation with the other therapeutic agent(s) or in separate formulations.

[0139] When the compounds of the present application are administered with other therapeutic agents, generally the amount of each component in the typical daily dose and in the typical dosage form can be reduced relative to the amount of each component that would be used if the components were administered alone, taking into account the additive or synergistic effects of the combination.

[0140] The compounds, or pharmaceutically acceptable salts, or hydrates thereof, or pharmaceutical compositions thereof, according to the present application are effective for preventing, treating or alleviating a PKR-mediated disease in a patient, in particular anemia.

[0141] In some embodiments, the compounds, or pharmaceutical compositions thereof, according to the present application are effective for preventing, treating or alleviating an anemia condition in a patient including, but not limited to, thalassemia, pyruvate deficiency anemia, anemia associated with myelodysplastic syndrome (MDS), sickle cell anemia, megaloblastic anemia, aplastic anemia, iron deficiency anemia, hemolytic anemia, sickle cell disease, hereditary non-spherocytic hemolytic anemia, hereditary spherocytosis, hereditary elliptocytosis, abetalipoproteinemia, paroxysmal nocturnal hemoglobinuria, acquired hemolysis, tumor-related anemia, or chronic disease-related anemia.

[0142] General synthetic procedures

[0143] For the purpose of illustrating the inventive technical solutions, the present application will be further described with the following examples. The following examples are only used to illustrate the specific implementation methods of the present application, so that those skilled in the art can understand the present application, but not used to limit the protection scope of the present application. In the specific implementation methods of the present application, the technical means or methods not specifically described are the conventional technical means or methods in the art.

[0144] Unless otherwise indicated, the definitions of substituents recited hereinabove apply equally to the following reaction schemes and examples, which serve to further illustrate the inventive technical solutions.

[0145] Those skilled in the art will appreciate that the chemical reactions described herein can be used to prepare other compounds of the present application according to synthetic routes known in the art and described herein. Additionally, other methods for preparing the compounds of the present application are set forth in the Examples below and / or can be apparent to those skilled in the art. For example, those non-exemplified compounds according to the present application can be successfully synthesized by modifying the methods described herein, such as by appropriately protecting interfering groups, by utilizing other known reagents and reactions, or by making routine modifications.

[0146] Unless otherwise indicated, all temperatures are set forth in degrees Celsius (C). Room temperature, unless otherwise indicated, means 15 °C to 30 °C; in some embodiments, room temperature is 20 °C to 30 °C. Reagents were purchased from commercial suppliers such as Aldrich Chemical Company, Arco Chemical Company and Alfa Chemical Company, and used without further purification. Unless otherwise indicated, general reagents were purchased from Shantou Xilong Chemical Factory, Guangdong Guanghua Chemical Reagent Factory, Guangzhou Chemical Reagent Factory, Tianjin Haoyu Chemicals Co., Ltd., Tianjin Fumian Chemical Reagent Factory, Wuhan Xinhua Yuan Science and Technology Development Co., Ltd., Qingdao Tenglong Chemical Reagent Co., Ltd., and Qingdao Haoyang Chemical Factory.

[0147] Anhydrous tetrahydrofuran, dioxane, toluene, diethyl ether were dried over sodium metal. Anhydrous dichloromethane and chloroform were dried over calcium hydride. Ethyl acetate, petroleum ether, n-hexane, N,N-dimethylacetamide and N,N-dimethylformamide were used as received.

[0148] The following reactions were generally performed under an atmosphere of nitrogen or argon at positive pressure or under anhydrous conditions in a dry box (unless otherwise indicated), reaction vessels were fitted with a septum and substrates were introduced via syringe. Glassware was oven- or flame-dried.

[0149] Chromatography columns were packed with silica gel. Silica gel (300-400 mesh) was purchased from Qingdao Haoyang Chemical Factory.

[0150] 1 H NMR spectra were recorded on a Bruker 400 MHz, 600 MHz or 599 MHz NMR spectrometer. 1 H NMR spectra were recorded in CDC13, DMSO-d6, CD3OD or acetone-d6 as solvent (in ppm) with TMS (0 ppm) or chloroform (7.26 ppm) as reference standard. When multiple peaks occur, the following abbreviations will be used: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broadened), br s (broadened singlet), dd (doublet of doublets), dt (doublet of triplets), qt (quartet of triplets). Coupling constants, J, are reported in Hertz (Hz).

[0151] Low resolution mass spectrometry (MS) data were acquired on an Agilent 6120 quadrupole HPLC-MS (chromatographic column type: Zorbax SB-C18, 2.1 x 30 mm, 3.5 micron, 6 min, flow rate 0.6 mL / min. Mobile phase: 5-95% (CH3CN with 0.1% formic acid) in (H2O with 0.1% formic acid), using electrospray ionization (ESI), with UV detection at 210 nm / 254 nm.

[0152] Pure compounds were analyzed using an Agilent 1260 pre-HPLC or Calesep pump 250 pre-HPLC (chromatographic column type: NOVASEP 50 / 80 mm DAC), with UV detection at 210 nm / 254 nm.

[0153] The following abbreviations or English words are used throughout the present application: EtOAc, EA ethyl acetate h hours DCM dichloromethane min minutes PE petroleum ether °C degrees Celsius MeOH methanol g grams DMSO dimethylsulfoxide mmol millimoles DMSO-d6 deuterated dimethylsulfoxide M, mol / L moles per liter mL milliliters DMF N,N-dimethylformamide DEAN, N-diethyl aniline CO2 carbon dioxide DAST diethylaminosulfur trifluoride EDCI 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride μM micromoles HOBt 1-hydroxybenzotriazole DIPEA N,N-diisopropylethylamine rt room temperature TEA triethylamine HCl hydrochloric acid BSA bovine serum albumin DTT dithiothreitol MgCl2 magnesium chloride KCl potassium chloride DIAD diisopropyl azodicarboxylate NCS N-chlorosuccinimide xantphos, Xantphos, Xant-Phos 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene Pd2(dba)3 tris(dibenzylideneacetone)dipalladium(0) NBS N-bromosuccinimide

[0154] Synthesis Scheme 1

[0155] The intermediate compound (I-1) can be synthesized according to the method of Synthesis Scheme 1. Wherein R 1with the definition as described in the application, X is halogen. Compound (1a) is reacted with sulfuryl chloride in a suitable solvent (such as tetrahydrofuran) to obtain compound (1b); compound (1b) is reacted with compound (1c) under suitable conditions (such as under the action of TEA) to obtain compound (1d); compound (1d) is reacted under acidic conditions (such as under the action of HCl) to obtain intermediate compound (I-1).

[0156] Synthetic scheme 2

[0157] Intermediate compound (I-2) can be synthesized according to the method of synthetic scheme 2. Wherein R 2 with the definition as described in the application, W is C 1-6 alkyl. Compound (2a) is reacted under the action of a fluorinating agent (such as DAST) to obtain compound (2b); compound (2b) is reacted with paraformaldehyde under suitable conditions (such as sodium methoxide) to obtain compound (2c); compound (2c) is subjected to ester hydrolysis under suitable conditions (such as LiOH) to obtain compound (I-2).

[0158] Synthetic scheme 3

[0159] Compound (I) can be synthesized according to the method of synthetic scheme 3. Wherein R 1 and R 2 with the definition as described in the application. Compound (I-1) is reacted with compound (I-2) under suitable conditions (such as under the action of EDCI, HOBt and DIPEA) to obtain compound (I).

[0160] The compounds, pharmaceutical compositions and applications thereof provided by the application are further illustrated in conjunction with the following examples. Examples

[0161] Intermediate 1-1: 2,3-dihydro-[1,4]dioxino[2,3-b]pyridine-7-sulfonyl chloride

[0162] Into a reaction flask was placed n-butyllithium (0.8 g, 12.50 mmol) under nitrogen protection, after the addition of n-butyllmagnesium (1.7 g, 12.26 mmol) and stirred for 10 minutes. The reaction solution was cooled to -10 °C, then a solution of 7-bromo-2,3-dihydro-[l,4]dioxino[2,3-b]pyridine (5.0 g, 23.14 mmol) in tetrahydrofuran (40 mL) was slowly added dropwise, and the stirring was continued at -10 °C for 1 h after the dropwise addition was completed. To the reaction solution was slowly added dropwise sulfuryl chloride (46.8 g, 347.1 mmol), and the stirring was continued at -10 °C for 30 min after the dropwise addition was completed. To the reaction solution was slowly added dropwise saturated ammonium chloride solution to quench the reaction, and the extraction was performed with dichloromethane (30 mL x 2), and the organic phase was dried over anhydrous sodium sulfate, then filtered, and the filtrate was concentrated under reduced pressure. The residue obtained was purified by column chromatography (PE:EtOAc (v:v) = 4: 1) to obtain a white solid product 4.4 g, with a yield of 80.6%.

[0163] The following intermediates were obtained according to the synthetic route of Intermediate 1-1

[0164] Intermediate 1-6: 5,6-dihydro-8H-imidazo[2,l-c][l,4]oxazine-2-sulfonyl chloride

[0165] First step: synthesis of ethyl 2-(3-oxomorpholinyl)acetate

[0166] Into a reaction flask was sequentially added morpholin-3-one (5.0 g, 49.46 mmol), cesium carbonate (32.23 g, 98.92 mmol), ethyl 2-bromoacetate (9.09 g, 54.41 mmol) and acetonitrile (150 mL), and the reaction was stirred at 60 °C for 16 hours. The extraction was performed under reduced pressure, the filtrate was rotary evaporated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 3 / 1) to obtain a colorless liquid title compound (6.7 g, 72.37%).

[0167] MS (ESI, pos.ion) m / z: 188.2 [M+H] + .

[0168] Second step: synthesis of 2-(3-oxomorpholinyl)acetamide

[0169] Ethyl 2-(3-oxomorpholinyl)acetate (6.7 g, 35.79 mmol) was dissolved in a 7M solution of ammonium methylate (51.13 mL, 357.9 mmol), and the reaction was performed at 60 °C for 14.5 hours. The rotary evaporation was performed under reduced pressure to obtain a white solid title compound (5.44 g, 96.10%).

[0170] MS (ESI, pos.ion) m / z: 159.2 [M+H]+ .

[0171] Step 3: Synthesis of 2-bromo-5,6-dihydro-8H-imidazo[2,1-c][1,4]oxazine

[0172] To a solution of 2-(3-oxomorpholine)acetamide (0.39 g, 2.47 mmol) in acetonitrile (5 mL) was added phosphorus tribromide (1.42 g, 4.94 mmol) and stirred at 90 °C for 3.5 h. The reaction was dropped into ice saturated sodium carbonate solution, extracted with ethyl acetate (30 mL x 2), washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 2 / 1) to give the title compound (0.19 g, 37.95%) as a yellow solid. MS (ESI, pos.ion) m / z: 203.1 [M+H] + ;

[0173] 1 H NMR (600 MHz, CDCl3) δ 6.84 (s, 1H), 4.79 (s, 2H), 4.06-4.04 (m, 2H), 4.01-3.98 (m, 2H).

[0174] Step 4: Synthesis of 5,6-dihydro-8H-imidazo[2,1-c][1,4]oxazine-2-sulfonyl chloride

[0175] Intermediate 1-6 was obtained by referring to the synthetic method of Intermediate 1-1, using 2-bromo-5,6-dihydro-8H-imidazo[2,1-c][1,4]oxazine as the substrate. Intermediate 1-7: 5-methoxypyridine-2-sulfonyl chloride

[0176] Step 1: Synthesis of 2-(benzylthio)-5-methoxypyridine

[0177] To a flask of 2-bromo-5-methoxypyridine (1.0 g, 5.32 mmol) in 1,4-dioxane (10 mL) was added DIPEA (1.38 g, 10.64 mmol) and stirred to dissolve, then added benzyl mercaptan (0.66 g, 5.32 mmol) and xantphos (0.31 g, 0.53 mmol), replaced by nitrogen, finally added Pd2dba3(0.24 g, 0.27 mmol), and warmed to 110 °C to react. TLC monitoring, the raw material was completely reacted, the reaction was stopped, the organic solvent was removed by reduced pressure concentration, extracted with ethyl acetate (20 mL x 3), separated, the organic phase was dried over anhydrous sodium sulfate, filtered, and dried, and then loaded onto a silica gel column for column chromatography (PE:EA (v:v) = 10:1) to give the title compound (1.15 g, 93.48%) as an orange oil.

[0178] MS (ESI, pos.ion) m / z: 232.0 [M+H] + ;

[0179] 1 H NMR (400 MHz, CDCl3) δ 8.24 (d, J = 2.6 Hz, 1H), 7.40 (d, J = 7.3 Hz, 2H), 7.30 (t, J = 7.4 Hz, 2H), 7.24 (t, J = 7.2 Hz, 1H), 7.11-7.07 (m, 2H), 4.40 (s, 2H), 3.84 (s, 3H).

[0180] Second step: synthesis of 5-methoxypyridine-2-sulfonyl chloride

[0181] To a flask of DCM (3 mL) was added 2-benzylsulfanyl-5-methoxypyridine (0.70 g, 3.03 mmol), NCS (2.02 g, 15.15 mmol) and benzyl alcohol (2.29 g, 21.21 mmol), and the reaction was continued at room temperature. TLC monitoring, the raw material was completely reacted, the reaction was stopped, concentrated by reduced pressure, dried and loaded onto a silica gel column for column chromatography (PE:EA (v:v) = 5:1) to give the title compound (0.45 g, 71.62%) as a yellow liquid.

[0182] Intermediate 2-1: 2-fluoro-3-hydroxy-2-phenylpropanoic acid

[0183] First step: synthesis of methyl 2-hydroxy-2-phenylacetate

[0184] To a solution of 2-hydroxy-2-phenylacetic acid (2.0 g, 13.15 mmol) in methanol (50 mL) was added p-toluenesulfonic acid monohydrate (0.22 g, 1.31 mmol) and stirred at 65 °C. The solvent was removed by rotary evaporation under reduced pressure, and the residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 20 / 1) to give the title compound (2.00 g, 91.56%) as a colorless liquid.

[0185] Second step: synthesis of methyl 2-fluoro-2-phenylacetate

[0186] Methyl 2-hydroxy-2-phenylacetate (2 g, 12.04 mmol) was dissolved in dichloromethane (50 mL) and cooled to 0 °C, then DAST (3.88 g, 24.08 mmol) was added dropwise. After the addition was completed, the reaction solution was allowed to warm to room temperature and stirred for 1 h. The reaction solution was quenched with ice water (30 mL) and extracted with dichloromethane (30 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (PE:EA (v:v) = 10:1) to give a yellowish oil (1.4 g, yield 69.1%).

[0187] 1 H NMR (400 MHz, CDC13) δ 7.55 - 7.35 (m, 5H), 5.82 (d, J = 47.6 Hz, 1H), 3.80 (s, 3H).

[0188] 19 F NMR (376 MHz, CDC13) δ -179.88 (s).

[0189] Third step: synthesis of methyl 2-fluoro-3-hydroxy-2-phenylpropanoate

[0190] Methyl 2-fluoro-2-phenylacetate (0.5 g, 2.97 mmol) was dissolved in dimethyl sulfoxide (10 mL), and sodium methoxide (0.16 g, 2.97 mmol) and paraformaldehyde (0.27 g, 8.91 mmol) were added sequentially at room temperature. After the addition was completed, the reaction solution was stirred at room temperature overnight. The reaction solution was diluted with ethyl acetate (20 mL) and washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (PE:EA (v:v) = 4:1) to give the product (0.2 g, yield 33.94%) as a white solid.

[0191] 1H NMR (400 MHz, CDC13) δ 7.53 (dt, J = 4.3, 2.4 Hz, 2H), 7.46 - 7.39 (m, 3H), 4.35 (dd, J = 30.0, 12.1 Hz, 1H), 4.04 (dd, J = 15.7, 12.7 Hz, 1H), 3.86 (s, 3H).

[0192] Fourth Step: Synthesis of 2-fluoro-3-hydroxy-2-phenylpropionic acid

[0193] To a solution of methyl 2-fluoro-3-hydroxy-2-phenylpropionate (0.8 g, 4.04 mmol) in water (10 mL) and methanol (20 mL) was added lithium hydroxide monohydrate (0.34 g, 8.08 mmol) and the reaction was stirred at room temperature overnight. The reaction was concentrated under reduced pressure and the residue was stirred in ethyl acetate (10 mL) for 10 min and then concentrated under reduced pressure. The crude product was dissolved in ethyl acetate (20 mL) and washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give the product as a white solid (0.6 g, 76.7% yield).

[0194] The following intermediates were synthesized according to the synthetic route of Intermediate 2-1.

[0195] Intermediate 2-11 2-fluoro-2-(3-fluorophenyl)-3-hydroxypropanoic acid

[0196] First Step: Synthesis of 3-fluoromandelic acid

[0197] A solution of 3-fluoroacetophenone (2 g, 14.48 mmol), ytterbium triflate (0.90 g, 1.45 mmol), selenium dioxide (3.21 g, 28.96 mmol) in 1,4-dioxane (48 mL) and water (16 mL) was heated to 90 °C and stirred overnight. After cooling to room temperature, the mixture was filtered through celite and the filter cake was washed with dichloromethane. Most of the dichloromethane was removed under reduced pressure and the remaining aqueous phase was adjusted to pH = 1 with 2% sodium hydroxide solution (20 mL). The mixture was stirred for 30 min and then extracted with dichloromethane (20 mL x 2). The aqueous phase was slowly adjusted to pH = 1 with 2 M dilute hydrochloric acid and then extracted with ethyl acetate (20 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give the title compound as a yellow solid (2 g, 81.19% yield).

[0198] MS (ESI, neg. ion) m / z: 168.8 [M-H] - ; 1H NMR (599 MHz, DMSO-d6) δ 12.78 (s, 1H), 7.40 (dd, J = 14.1, 7.8 Hz, 1H), 7.28 (d, J = 7.6 Hz, 1H), 7.23 (d, J = 10.0 Hz, 1H), 7.13 (td, J = 8.7, 2.0 Hz, 1H), 5.09 (s, 1H).

[0199] Synthesis of 2-fluoro-2-(3-fluorophenyl)-3-hydroxypropanoic acid

[0200] The target intermediate 2-11 was obtained by referring to the synthetic route of Example 2-1 using 3-fluoromandelic acid as the reactant.

[0201] Intermediate 2-12: 2-(4-chloro-2-fluorophenyl)-2-fluoro-3-hydroxypropanoic acid

[0202] First step: Synthesis of ethyl 2-(4-chloro-2-fluorophenyl)-2-fluoroacetate

[0203] Ethyl bromofluoroacetate (1.0 g, 5.41 mmol) was dissolved in 1,4-dioxane (20 mL), and (4-chloro-2-fluorophenyl)boronic acid (1.89 g, 10.82 mmol), triphenylphosphine (0.14 g, 0.54 mmol), nickel(II) trifluoromethanesulfonate (0.10 g, 0.27 mmol), 4,4'-di-tert-butyl-2,2'-bipyridine (0.07 g, 0.27 mmol), and potassium carbonate (0.24 g, 16.23 mmol) were added. The reaction was stirred at 80°C under nitrogen protection until completion. After the reaction was completed, the reaction was quenched by adding an appropriate amount of water, extracted with ethyl acetate (3 x 25 mL), and the organic layer was collected and washed with saturated brine (1 x 30 mL) and dried over anhydrous sodium sulfate. The residue was concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 4 / 1) to obtain the title compound (1.15 g, 90.67%) as a transparent oil.

[0204] Synthesis of 2-(4-chloro-2-fluorophenyl)-2-fluoro-3-hydroxypropanoic acid

[0205] The intermediate 2-12 was obtained by referring to the synthetic method of Intermediate 2-1 using ethyl 2-(4-chloro-2-fluorophenyl)-2-fluoroacetate as the reactant.

[0206] The following intermediates were synthesized by referring to the synthetic route of Intermediate 2-12.

[0207] Intermediate 2-19 2-Fluoro-3-hydroxy-2-(4-(methylsulfonyl)phenyl)propanoic acid

[0208] First Step: Synthesis of ethyl 2-fluoro-2-(4-(methylthio)phenyl)acetate

[0209] Ethyl 2-fluoro-2-(4-(methylthio)phenyl)acetate was synthesized by referring to the first step of the synthesis of Intermediate 2-12, using (4-(methylthio)phenyl)boronic acid as the starting material. 1 H NMR (400 MHz, CDC13) δ 7.39 (dd, J = 8.3, 1.4 Hz, 2H), 7.28 (d, J = 8.2 Hz, 2H), 5.74 (d, J = 47.7 Hz, 1H), 4.32 - 4.20 (m, 2H), 2.51 (s, 3H), 1.28 (s, 3H). 19 F NMR (376 MHz, CDC13) δ -178.48.

[0210] Second Step: Synthesis of ethyl 2-fluoro-2-(4-(methylsulfonyl)phenyl)acetate

[0211] Ethyl 2-fluoro-2-(4-(methylsulfonyl)phenyl)acetate was synthesized by referring to the first step of the synthesis of Intermediate 2-12, using (4-(methylthio)phenyl)boronic acid as the starting material.

[0212] MS (ESI, pos.ion) m / z: 261.1 [M+H] + .

[0213] Third and Fourth Steps: Synthesis of 2-fluoro-3-hydroxy-2-(4-(methylsulfonyl)phenyl)propanoic acid

[0214] Intermediate 2-19 2-Fluoro-3-hydroxy-2-(4-(methylsulfonyl)phenyl)propanoic acid

[0215] Intermediate 2-20 2-(2,3-Dihydro-1H-inden-5-yl)-2-fluoro-3-hydroxypropanoic acid

[0216] First step: synthesis of 2-(2,3-dihydro-1H-inden-5-yl)-2-((trimethylsilyl)oxy)acetonitrile

[0217] To a solution of indan-5-carbaldehyde (2 g, 13.68 mmol) in dichloromethane (50 mL) was added zinc iodide (0.87 g, 2.74 mmol), and trimethylsilyl cyanide (2.57 mL, 20.52 mmol) was added dropwise under ice-bath. The reaction was continued under ice-bath for 3.5 hours. Water (50 mL) was added to the reaction, and stirred at room temperature for 50 minutes. The organic phase was collected after standing and partitioning. The aqueous phase was further extracted with dichloromethane (30 mL). The combined organic phase was washed with water (40 mL), saturated brine (40 mL) successively, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10 / 1) to give the title compound (2.7 g, 80.43%) as yellow oil.

[0218] Second step: synthesis of methyl 2-(2,3-dihydro-1H-inden-5-yl)-2-hydroxyacetate

[0219] To a solution of 2-(2,3-dihydro-1H-inden-5-yl)-2-((trimethylsilyl)oxy)acetonitrile (2.67 g, 10.88 mmol) in methanol (10 mL) was added dropwise a solution of hydrogen chloride (8.16 mL, 32.64 mmol) in 1,4-dioxane. The reaction was stirred at 65 °C for 5 hours. TLC monitoring showed that the starting material had been completely reacted, and the reaction was stopped. The solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10 / 1) to give the title compound (1.80 g, 80.21%) as light yellow oil.

[0220] 1 H NMR (400 MHz, CDCl3) δ 7.26 (s, 1H), 7.21 (d, J = 7.7 Hz, 1H), 7.16 (dd, J = 7.6, 1.7 Hz, 1H), 5.14 (d, J = 5.7 Hz, 1H), 3.76 (s, 3H), 3.37 (d, J = 5.7 Hz, 1H), 2.93-2.87 (m, 4H), 2.09-2.05 (m, 2H).

[0221] Third to fifth steps: synthesis of 2-(2,3-dihydro-1H-inden-5-yl)-2-fluoro-3- hydroxypropanoic acid

[0222] The intermediate compound 2-20 was obtained according to the synthesis method of the intermediate 2-1.

[0223] Intermediate 2-21 2-fluoro-3-hydroxy-2-(1-methyl-1H-indazol-5-yl)propanoic acid

[0224] Intermediate 2-21 was obtained by using 1-methyl-1H-indazole-5-carboxaldehyde as raw material, referring to the synthetic method of intermediate 2-20.

[0225] Intermediate 2-22 (R)-2-fluoro-3-hydroxy-2-phenylpropan-1-one

[0226] Intermediate 2-1 as raw material, referring to the synthetic method of example 2 of patent CN117603204A.

[0227] 1 HNMR (400 MHz, DMSO-d6) δ 7.50-7.36 (m, 5H), 4.14 (dd, J = 31.6, 12.4 Hz, 1H), 3.80 (dd, J = 17.3, 12.4 Hz, 1H).

[0228] Example 1 and Example 2: (R)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7- yl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3-hydroxy-2- phenylpropan-1-one and (S)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl)sulfonyl)- 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3-hydroxy-2-phenylpropan-1- one

[0229] First step: synthesis of tert-butyl 5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl)sulfonyl)- 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate

[0230] Intermediate 2-1 as raw material, referring to the synthetic method of example 2 of patent CN117603204A. +.

[0231] Step 2: Synthesis of 7-((3,4,5,6-tetrahydro pyrrolo[3,4-c]pyrrol-2(lH)-yl) sulfonyl)- 2,3-dihydro-[l,4]dioxino[2,3-b]pyridine

[0232] Tert-butyl 5-((2,3-dihydro-[l,4]dioxino[2,3-b]pyridin-7-yl)sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrole-2(lH)-carboxylate (1.9 g, 4.64 mmol) was dissolved in dichloromethane (50 mL) at room temperature, and trifluoroacetic acid (18.4 g, 161.55 mmol) was added. The reaction was stirred at room temperature overnight. The reaction was concentrated under reduced pressure, and most of the organic solvent was removed. The residue was diluted with water and adjusted to weak alkaline with saturated sodium bicarbonate solution. The product was extracted with dichloromethane (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (DCM:MeOH (v:v) = 10:1) to give the product as a white solid, 1.5 g, in 99% yield.

[0233] Step 3: Synthesis of l-(5-((2,3-dihydro-[l,4]dioxino[2,3-b]pyridin-7-yl)sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)-2-fluoro-3-hydroxy-2-phenylpropan-l-one

[0234] To the reaction flask, 2-fluoro-3-hydroxy-2-phenylpropanoic acid (0.33 g, 1.79 mmol), 7-((3,4,5,6-tetrahydro pyrrolo[3,4-c]pyrrol-2(lH)-yl)sulfonyl)-2,3-dihydro- [l,4]dioxino[2,3-b]pyridine (0.68 g, 1.97 mmol), EDCI (0.45 g, 2.33 mmol), and HOBt (0.36 g, 2.69 mmol) were added successively. DMF (15 mL) and DIPEA (0.93 g, 7.16 mmol) were added successively under nitrogen protection. The reaction was stirred at room temperature overnight. The reaction was quenched with water and extracted with dichloromethane (20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (DCM:MeOH (v:v) = 20:1) to give the product as a yellow solid, 0.4 g, in 47.0% yield.

[0235] Step 4: Chromatographic preparation of resolution

[0236] The third step product was separated by chiral-preparative-HPLC (Chiral column: Chiralpak OD, 10 mm x 250 mm, 5 um; mobile phase A: 30% CO2; mobile phase B: 70% MeOH + 0.1% DEA; flow rate: 5 mL / min; detector UV wavelength 210 nm, 254 nm) to give two enantiomers as a light yellow solid (S)-isomer (Example 2) (60 mg, yield 15%, retention time RT (min) = 10.5 min) and a light yellow solid (R)-isomer (Example 1) (52 mg, yield 13%, retention time RT (min) = 12.5 min), respectively.

[0237] Example 1: 1 H NMR (400 MHz, DMSO-d6) δ 8.14 (d, J = 2.2 Hz, 1H), 7.61 (d, J = 2.2 Hz, 1H), 7.44 - 7.35 (m, 5H), 5.30 (t, J = 5.8 Hz, 1H), 4.50 (dd, J = 4.9, 3.0 Hz, 2H), 4.43 (d, J = 15.5 Hz, 1H), 4.32 (dd, J = 4.8, 3.0 Hz, 2H), 4.18 - 3.96 (m, 7H), 3.94 (d, J = 16.6 Hz, 1H), 3.73 - 3.65 (m, 1H).

[0238] 19 F NMR (376 MHz, DMSO) δ -170.42 (s). MS (ESI, pos.ion) m / z: 476.1 [M+H] + .

[0239] Example 2: 1 H NMR (400 MHz, DMSO-d6) δ 8.14 (d, J = 2.1 Hz, 1H), 7.61 (d, J = 2.2 Hz, 1H), 7.48 - 7.33 (m, 5H), 5.30 (t, J = 5.8 Hz, 1H), 4.50 (dd, J = 4.9, 2.9 Hz, 2H), 4.43 (d, J = 15.3 Hz, 1H), 4.32 (dd, J = 4.7, 3.0 Hz, 2H), 4.18 - 3.99 (m, 7H), 3.95 (d, J = 15.9 Hz, 1H), 3.75 - 3.62 (m, 1H).

[0240] 19 F NMR (376 MHz, DMSO) δ -170.43 (s). MS (ESI, pos.ion) m / z: 476.1 [M+H] + .

[0241] Example 3 (R)-2-fluoro-1-(5-((2-fluorophenyl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4- c]pyrrol-2(1H)-yl)-3-hydroxy-2-phenylpropan-1-one

[0242] First Step: Synthesis of tert-butyl 5-((2-fluorophenyl)sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate

[0243] To a solution of 2-fluorobenzenesulfonyl chloride (0.1 g, 0.51 mmol) and tert-butyl 2,3,4,6-tetrahydro-1H-pyrrolo[3,4-c]pyrrole-5-carboxylate 4-methylbenzenesulfonate (0.20 g, 0.51 mmol) in dichloromethane (10 mL), triethylamine (0.15 g, 1.53 mmol) was added, and the mixture was stirred at room temperature overnight. The reaction was quenched with water (20 mL), and the mixture was separated. The aqueous phase was extracted with dichloromethane (20 mL x 2), and the combined organic phase was washed with saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (PE:EA (v:v) = 5:1) to give the product as a white solid (0.13 g, 68.67% yield). MS (ESI, pos.ion) m / z: 391.2 [M+Na] + .

[0244] Second Step: Synthesis of 2-((2-fluorophenyl)sulfonyl)-1,2,3,4,5,6-hexahydropyrrolo[3,4- c]pyrrole hydrochloride

[0245] To a solution of tert-butyl 5-((2-fluorophenyl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4- c]pyrrole-2(1H)-carboxylate (0.13 g, 0.35 mmol) in ethyl acetate (10 mL), a solution of hydrochloric acid in ethyl acetate (0.13 g, 3.5 mmol) was added, and the mixture was stirred at room temperature overnight. The reaction was concentrated under reduced pressure to give the product as a white solid (95 mg, 88.34% yield).

[0246] Third Step: Synthesis of (R)-2-fluoro-1-(5-((2-fluorophenyl)sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-3-hydroxy-2-phenylpropan-1-one

[0247] To a reaction flask was added 2-((2-fluorophenyl)sulfonyl)-1,2,3,4,5,6- hexahydropyrrolo[3,4-c]pyrrole hydrochloride (0.095 g, 0.31 mmol), (R)-2-fluoro-3- hydroxy-2-phenylpropanoic acid (0.057 g, 0.31 mmol), EDCI (0.071 g, 0.37 mmol) and HOBt (0.050 g, 0.37 mmol), followed by DMF (5 mL) and DIPEA (0.16 g, 1.24 mmol) under nitrogen protection. After addition, the reaction mixture was stirred at room temperature overnight. The reaction mixture was quenched with water and extracted with dichloromethane (20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (PE:EtOAc (v:v) = 2:1) to give the product as a white solid (35 mg, 25.8% yield).

[0248] MS (ESI, pos.ion) m / z: 434.9 [M+H] + ;

[0249] 1 H NMR (600 MHz, DMSO-d6) δ 7.80 (t, J = 6.8 Hz, 1H), 7.75 - 7.72 (m, 1H), 7.49 - 7.45 (m, 1H), 7.44 - 7.39 (m, 5H), 7.38 - 7.36 (m, 1H), 5.29 (t, J = 6.1 Hz, 1H), 4.46 (d, J = 14.6 Hz, 1H), 4.21 - 4.02 (m, 7H), 3.98 (d, J = 14.8 Hz, 1H), 3.69 ((ddd, J = 19.9, 12.3, 6.0 Hz, 1H). 19 F NMR (564 MHz, DMSO-d6) δ -108.61 (s), -170.33 (s).

[0250] Example 68 (R)-2-fluoro-3-hydroxy-2-phenyl-1-(5-(pyridin-2-ylsulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)propan-1-one

[0251] First Step: Synthesis of tert-butyl 5-(pyridin-2-ylsulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate

[0252] To a reaction flask was added tert-butyl 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-2(lH)- carboxylate p-toluenesulfonic acid salt (0.4 g, 1.05 mmol), dichloromethane (5 mL), triethylamine (0.27 g, 2.63 mmol) and pyridine-2-sulfonyl chloride (0.20 g, 1.10 mmol) sequentially, stirred at room temperature for 4 h; concentrated under reduced pressure, the residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 3 / 1) to give the title compound (0.17 g, 46.26%) as a white solid. MS (ESI, pos.ion) m / z: 352.2 [M+H] + .

[0253] Second Step: Synthesis of 2-(pyridin-2-ylsulfonyl)-l,2,3,4,5,6-hexahydropyrrolo[3,4- c]pyrrole dihydrochloride

[0254] To a solution of tert-butyl 5-(pyridin-2-ylsulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4- c]pyrrole-2(lH)-carboxylate (0.16 g, 0.46 mmol) in 1,4-dioxane was added a solution of HC1 (2.30 mL, 9.2 mmol, 4.0 M) in 1,4-dioxane, stirred at room temperature for 11.5 h. Rotated to dryness under reduced pressure to give the title compound (0.15 g, 100%) as a pink solid, which was directly used in the next step.

[0255] Third Step: Synthesis of (R)-2-fluoro-3-hydroxy-2-phenyl-l-(5-(pyridin-2-ylsulfonyl)- 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)propan-l-one

[0256] To a reaction flask was added (R)-2-fluoro-3-hydroxy-2-phenylpropanoic acid (0.080 g, 0.43 mmol), 2-(pyridin-2-ylsulfonyl)-l,2,3,4,5,6-hexahydropyrrolo[3,4-c]pyrrole dihydrochloride (0.15 g, 0.45 mmol), EDCI (0.11 g, 0.56 mmol) and HOBt (0.087 g, 0.65 mmol) sequentially, followed by DMF (5 mL) and DIPEA (0.22 g, 1.72 mmol), stirred at room temperature for 22.5 h. Added water (20 mL) to the reaction system, extracted with ethyl acetate (20 mL x 2), collected the organic phase, washed with water (20 mL), saturated brine (20 mL) sequentially, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, the residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 1 / 1) to give the title compound (0.092 g, 50.73%) as a white solid. MS (ESI, pos.ion) m / z: 418.2 [M+H]+ ;

[0257] 1 H NMR (600 MHz, DMSO-d6) δ 8.72 - 8.71 (m, 1H), 8.11 - 8.08 (m, 1H), 7.95 - 7.93 (m, 1H), 7.69 - 7.67 (m, 1H), 7.46 - 7.35 (m, 5H), 5.30 (t, J = 6.1 Hz, 1H), 4.44 (d, J = 15.0 Hz, 1H), 4.26 - 4.07 (m, 7H), 3.97 (d, J = 15.1 Hz, 1H), 3.69 (ddd, J = 20.1, 12.3, 6.0 Hz, 1H). 19 F NMR (564 MHz, DMSO-d6) δ -170.26.

[0258] Example 83 2-(4-chloro-2-fluorophenyl)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7- yl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3- hydroxypropan-1-one

[0259] Example 83 2-(4-chloro-2-fluorophenyl)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7- yl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3- hydroxypropan-1-one + ;

[0260] 1H NMR (599 MHz, DMSO-d6) δ 8.17 (d, J = 2.2 Hz, 1H), 7.63 (d, J = 2.2 Hz, 1H), 7.51 - 7.47 (m, 2H), 7.35 (dd, J = 8.4, 2.1 Hz, 1H), 5.42 (t, J = 6.1 Hz, 1H), 4.54 - 4.51 (m, 2H), 4.41 (d, J = 15.1 Hz, 1H), 4.36 - 4.33 (m, 2H), 4.15 - 4.02 (m, 9H).

[0261] The corresponding sulfonyl chloride starting material and (R)-2-fluoro-3-hydroxy-2- phenylpropanoic acid were used as starting materials to give the corresponding end products according to the synthetic procedure described in Example 3. See the table below for specific information.

[0262] The corresponding sulfonyl chloride starting material and (R)-2-fluoro-3-hydroxy-2- phenylpropanoic acid were used as starting materials to give the corresponding end products according to the synthetic procedure described in Example 3. See the table below for specific information.

[0263] Example 81 (S)-2-(2,6-difluorophenyl)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7- yl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3- hydroxypropan-1-one and

[0264] Example 82 (R)-2-(2,6-difluorophenyl)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7- yl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3-hydroxypropan-1- one

[0265] Chromatographic preparative resolution of sample of example 105

[0266] Chromatographic column: Chiralcel OD-H column 5um*10mm*250mm (waters supercritical fluid chromatography), sample dissolved in methanol (with a small amount of formic acid to aid dissolution), 8 mL / min column temperature 35 °C back pressure 100 bar; isocratic 45% methanol + 55% carbon dioxide, fraction I (7-8.05 min), fraction II (8.25-9.25 min);

[0267] Example 81 fraction I

[0268] MS (ESI, pos.ion) m / z: 512.2 [M+H] + ;

[0269] 1 H NMR (400 MHz, CDC13) δ 8.28 (d, J = 2.2 Hz, 1H), 7.58 (d, J = 2.2 Hz, 1H), 7.39 - 7.35 (m, 1H), 6.95 (t, J = 9.2 Hz, 2H), 4.59 - 4.52 (m, 2H), 4.49 (d, J = 14.8 Hz, 1H), 4.36 - 4.32 (m, 2H), 4.32 - 4.27 (m, 3H), 4.25 (s, 1H), 4.16 (s, 2H), 4.08 (s, 2H), 4.00 (d, J = 15.1 Hz, 1H). 19 F NMR (376 MHz, CDC13) δ -109.36, -162.91.

[0270] ee value: 100%

[0271] Example 82 fraction II

[0272] MS (ESI, pos.ion) m / z: 512.2 [M+H] + ;

[0273] 1H NMR (400 MHz, CDC13) δ 8.28 (d, J = 2.1 Hz, 1H), 7.58 (d, J = 2.2 Hz, 1H), 7.42 - 7.34 (m, 1H), 6.95 (t, J = 9.2 Hz, 2H), 4.57 - 4.51 (m, 2H), 4.49 (d, J = 13.9 Hz, 1H), 4.35 - 4.32 (m, 2H), 4.32 - 4.27 (m, 3H), 4.25 (s, 1H), 4.16 (s, 2H), 4.08 (s, 2H), 4.00 (d, J = 15.2 Hz, 1H). 19 F NMR (376 MHz, CDC13) δ -109.30, -162.71.

[0274] ee value: 98.44%

[0275] Example 89 (S)-2-(2,3-fluorophenyl)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7- yl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3- hydroxypropan-1-one and

[0276] Example 90 (R)-2-(2,3-fluorophenyl)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7- yl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3- hydroxypropan-1-one

[0277] Chromatographic preparation of the sample of Example 106 for separation

[0278] Chromatographic column: Chiralpak IC column 5um*10mm*250mm (waters supercritical fluid chromatography), 8 mL / min column temperature 35 °C back pressure 100 bar; isocratic 85% methanol + 15% carbon dioxide, collect fraction I (11.45 min - 12.55 min) and peak fraction II (12.65 min - 14.4 min).

[0279] Example 89 fraction I

[0280] MS (ESI, pos.ion) m / z: 512.0 [M+H] + ;

[0281] 1H NMR (400 MHz, Chloroform-d) δ 8.28 (s, 1H), 7.58 (s, 1H), 7.24 (dd, J = 26.7, 11.3 Hz, 3H), 4.54 (s, 2H), 4.41 (d, J = 14.6 Hz, 1H), 4.37 - 4.27 (m, 4H), 4.19 - 3.99 (m, 5H), 3.73 (d, J = 14.9 Hz, 1H), 3.25 (d, J = 16.3 Hz, 1H). 19 F NMR (376 MHz, Chloroform-d) δ -136.07, -138.48, -168.38.

[0282] ee value: 100%

[0283] Example 90 Component II

[0284] MS (ESI, pos.ion) m / z: 511.9 [M+H] + ;

[0285] 1 H NMR (400 MHz, Chloroform-d) δ 8.28 (s, 1H), 7.58 (s, 1H), 7.23 (q, J = 8.3 Hz, 3H), 4.54 (s, 2H), 4.41 (d, J = 13.9 Hz, 1H), 4.36 - 4.26 (m, 4H), 4.20 - 4.00 (m, 5H), 3.72 (d, J = 14.8 Hz, 1H), 3.23 (s, 1H).

[0286] 19 F NMR (376 MHz, Chloroform-d) δ -136.04 -138.46, -168.43.

[0287] ee value: 93.27%

[0288] Example 92 (S)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl)sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3-hydroxy-2-(p-tolyl)propan-1-one and

[0289] Example 93 (R)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl)sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3-hydroxy-2-(p-tolyl)propan-1-one

[0290] Chromatographic preparative resolution was performed on the sample of Example 107

[0291] Chromatographic column: Chiralcel OJ column 5um*10mm*250mm (waters supercritical fluid chromatography), 5 mL / min Column temperature 35 °C Back pressure 100 bar; Isocratic 80% methanol + 20% carbon dioxide, collect fraction I (15 min - 17.5 min) and fraction II (20.5 min - 23.7 min).

[0292] Example 92 Fraction I

[0293] MS (ESI, pos.ion) m / z: 490.0 [M+H] + ;

[0294] 1 H NMR (400 MHz, Chloroform-d) δ 8.28 (d, J = 2.2 Hz, 1H), 7.57 (d, J = 2.2 Hz, 1H), 7.31 (d, J = 7.9 Hz, 2H), 7.23 (d, J = 7.9 Hz, 2H), 4.56 - 4.52 (m, 2H), 4.44 (d, J = 15.5 Hz, 1H), 4.36 - 4.30 (m, 3H), 4.24 - 4.00 (m, 6H), 3.77 (t, J = 17.4 Hz, 2H), 2.38 (s, 3H). 19 F NMR (376 MHz, Chloroform-d) δ -172.73. Enantiomeric excess: 100%

[0295] Example 93 Fraction II

[0296] MS (ESI, pos.ion) m / z: 490.0 [M+H] + ;

[0297] 1 H NMR (400 MHz, Chloroform-d) δ 8.28 (d, J = 2.1 Hz, 1H), 7.57 (d, J = 2.2 Hz, 1H), 7.31 (d, J = 8.1 Hz, 2H), 7.23 (d, J = 8.0 Hz, 2H), 4.54 (dd, J = 5.4, 2.9 Hz, 2H), 4.44 (d, J = 15.2 Hz, 1H), 4.34 (t, J = 4.2 Hz, 3H), 4.24 - 4.01 (m, 6H), 3.79 (dd, J = 30.9, 13.3 Hz, 2H), 2.38 (s, 3H).

[0298] 19F NMR (376 MHz, Chloroform-d) δ -172.73. ee value: 100%

[0299] Example 97 (S)-2-(2,5-difluorophenyl)-1-(5-((2,3-dihydroxy-[1,4]dioxino[2,3-b]pyridin-7- yl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3-hydroxypropan-1- one and

[0300] Example 98 (R)-2-(2,5-difluorophenyl)-1-(5-((2,3-dihydroxy-[1,4]dioxino[2,3-b]pyridin-7- yl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3-hydroxypropan-1- one

[0301] Chromatographic preparation of the sample as in example 108 for the separation

[0302] Chromatographic column: Chiral IC column 5um*10mm*250mm (waters supercritical fluid chromatography), sample dissolved in DMSO, 8 mL / min column temperature 35°C back pressure 100 bar; isocratic 85% methanol + 15% carbon dioxide component I (12.3 min - 13.6 min), component II (14 min - 15.8 min).

[0303] Example 97 component I

[0304] 1 H NMR (400 MHz, CDCl3) δ 8.26 (d, J = 2.0 Hz, 1H), 7.56 (d, J = 2.0 Hz, 1H), 7.19 (t, J = 6.5 Hz, 1H), 7.12 - 7.02 (m, 2H), 4.58 - 4.47 (m, 2H), 4.45 - 3.90 (m, 11H), 3.67 (d, J = 20.9 Hz, 1H). ee value: 91.24%

[0305] Example 98 component II

[0306] 1H NMR (400 MHz, CDC13) δ 8.26 (d, J = 2.1 Hz, 1H), 7.56 (d, J = 2.1 Hz, 1H), 7.19 (t, J = 6.5 Hz, 1H), 7.08 (dd, J = 6.9, 5.5 Hz, 2H), 4.52 (dd, J = 5.0, 3.1 Hz, 2H), 4.44 - 3.90 (m, 11H), 3.68 (d, J = 14.6 Hz, 1H). ee value: 90.62%.

[0307] Example 109: (R)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl)sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3-hydroxy-2-(phenyl-d5)propan-1-one

[0308] First Step: Synthesis of ethyl 2-fluoro-2-(phenyl-d5)acetate

[0309] Ethyl 2-fluoro-2-(phenyl-d5)acetate (0.32 ml, 2.70 mmol) was dissolved in 1,4-dioxane (10 mL), added (phenyl-d5)boronic acid (0.34 g, 2.70 mmol), triphenylphosphine (0.07 g, 0.27 mmol), nickel(II) trifluoromethanesulfonate (0.05 g, 0.14 mmol), 4,4'-di-tert-butyl-2,2'-bipyridine (0.04 g, 0.14 mmol), potassium carbonate (1.12 g, 8.10 mmol), and stirred under nitrogen protection at 100 °C until the reaction was complete. The reaction was quenched with water (20 mL) and extracted with ethyl acetate (3 x 20 mL), and the organic layer was collected and washed with saturated brine (20 mL) and dried over anhydrous sodium sulfate. The residue was concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 15 / 1) to give the title compound (0.4 g, 79.05%) as a transparent oil.

[0310] 1 H NMR (400 MHz, CDC13) δ 5.80 (d, J = 47.8 Hz, 1H), 4.36 - 4.18 (m, 2H), 1.29 (t, J = 7.1 Hz, 3H).

[0311] 19 F NMR (376 MHz, CDC13) δ -179.97.

[0312] Second Step: Synthesis of ethyl 2-fluoro-3-hydroxy-2-(phenyl-d5)propanoate

[0313] Ethyl 2-fluoro-2-(phenyl-d5)acetate (3.40 g, 18.16 mmol) was dissolved in DMF (20 mL), and paraformaldehyde (1.64 g, 54.48 mmol), sodium ethoxide (1.07 mL, 2.72 mmol) were added. The reaction was stirred at room temperature. After the reaction was completed, water (40 mL) was added to quench the reaction, and then extracted with ethyl acetate (3 x 30 mL). The organic layer was washed with saturated brine (30 mL) and dried over anhydrous sodium sulfate. After filtration, the organic solvent was removed under reduced pressure. The residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 4 / 1) to give the title compound (2.9 g, 73.50%) as a transparent oil.

[0314] 1 H NMR (400 MHz, CDC13) δ 4.41 - 4.27 (m, 3H), 4.03 (ddd, J = 15.8, 12.6, 5.3 Hz, 1H), 2.33 - 2.26 (m, 1H), 1.32 (t, J = 7.1 Hz, 3H). 19 F NMR (376 MHz, CDC13) δ -171.67.

[0315] Third Step: Synthesis of 2-fluoro-3-hydroxy-2-(phenyl-d5)propanoic acid

[0316] Ethyl 2-fluoro-3-hydroxy-2-(phenyl-d5)propanoate (2.60 g, 11.97 mmol) was dissolved in THF (20 mL), and lithium hydroxide monohydrate (0.72 g, 29.93 mmol) in water (4 mL) was added. The reaction was stirred at room temperature until the reaction was completed. After the reaction was completed, the organic solvent was removed under reduced pressure. The residue was dissolved in water (10 mL), and the pH was adjusted to 1-2 with dilute hydrochloric acid (1 M). The reaction was extracted with ethyl acetate (3 x 30 mL), and then washed with saturated brine (20 mL) and dried over anhydrous sodium sulfate. The white solid of the title compound was obtained after concentration under reduced pressure (1.45 g, 64.04%). MS (ESI, neg. ion) m / z: 188.1 [M-H] - .

[0317] Fourth Step: Synthesis of (R)-2-fluoro-3-hydroxy-2-(phenyl-d5)propanoic acid

[0318] Dissolve 2-fluoro-3-hydroxy-2-(phenyl-d5)propanoic acid (1.38 g, 7.29 mmol) in ethanol (15 mL), stir at 80 °C until dissolved, then add (1R,2R)-2-amino-1-(4-nitrophenyl)propane-1,3-diol (0.70 g, 3.28 mmol), stir at 80 °C until the reaction is complete. After the reaction is cooled to room temperature naturally, continue to cool in an ice bath, and a white solid is precipitated. Filter, wash the filter cake with ethanol, collect the filter cake, add an appropriate amount of water to dissolve and dilute, then adjust the pH to 1 with dilute hydrochloric acid (1 M), stir for 10 minutes, then extract with ethyl acetate (3 x 25 mL), and dry the combined organic layers over anhydrous sodium sulfate. Concentrate under reduced pressure to obtain the title compound (0.63 g, 45.6%) as a white solid. MS (ESI, neg. ion) m / z: 188.1 [M-H] - .

[0319] Fifth step: synthesis of (R)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3-hydroxy-2-(phenyl-d5)propan-1-one

[0320] Dissolve 7-((3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)sulfonyl)-2,3-dihydro-[1,4]dioxino-[2,3-b]pyridine hydrochloride (0.30 g, 0.78 mmol) in THF (10 mL), add DIPEA (0.39 mL, 2.34 mmol), HOBt (0.16 g, 1.17 mmol), EDCI (0.22 g, 1.17 mmol), (R)-2-fluoro-3-hydroxy-2-(phenyl-d5)propanoic acid (0.15 g, 0.78 mmol), protect with nitrogen, and stir at room temperature until the reaction is complete. After the reaction is complete, quench the reaction with an appropriate amount of water, extract with ethyl acetate (3 x 25 mL), take the organic layer, wash with saturated brine (25 mL), and dry over anhydrous sodium sulfate. Filter, concentrate under reduced pressure, and purify the residue by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1 / 4) to obtain the title compound (0.26 g, 68.96%) as a white solid. MS (ESI, pos. ion) m / z: 481.0 [M+H] + ;

[0321] 1H NMR (400MHz, DMSO-d6) δ8.12(s,1H),7.58(s,1H),5.27(t,J=6.4Hz,1H),4.47(s,2H),4.41( d,J=14.9Hz,1H),4.30(s,2H),4.06(t,J=14.3Hz,8H),3.66(ddd,J=19.4,12.4,5.8Hz,1H).

[0322] 19 F NMR(376MHz,DMSO-d6)δ-170.42.

[0323] Example 110: (R)-2-fluoro-3-hydroxy-2-(phenyl-d5)-1-(5-(pyridin-2-ylsulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrolo-2(1H)-yl)propane-1-one

[0324] 2-(pyridine-2-sulfonyl)-1,2,3,4,5,6-hexahydropyrrolo[3,4-c]pyrrole dihydrochloride (0.30 g, 0.93 mmol) was dissolved in tetrahydrofuran (10 mL), and DIPEA (0.46 mL, 2.79 mmol), HOBt (0.19 g, 1.40 mmol), EDCI (0.27 g, 1.40 mmol), and (R)-2-fluoro-3-hydroxy-2-(phenyl-d5)propionic acid (0.18 g, 0.93 mmol) were added. Under nitrogen protection, the mixture was stirred at room temperature until the reaction was complete. After the reaction was complete, an appropriate amount of water was added to quench the reaction, and the mixture was extracted with ethyl acetate (3 × 25 mL). The organic layer was collected, washed with saturated brine (25 mL), and dried over anhydrous sodium sulfate. The residue was filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1 / 3) to give the title compound as a white solid (0.25 g, 63.97%). MS (ESI, pos.ion) m / z: 423.0 [M+H] + ;

[0325] 1 H NMR (400MHz, DMSO-d6) δ8.68(d,J=4.6Hz,1H),8.07(t,J=8.0Hz,1H),7.91(d,J=8.0Hz,1H),7.65(t,J=6.3Hz,1H),5.29( t,J=6.2Hz,1H),4.42(d,J=15.4Hz,1H),4.21–4.05(m,7H),3.94(d,J=15.7Hz,1H),3.67(ddd,J=19.2,12.4,5.8Hz,1H). 19F NMR (376 MHz, DMSO-d6) δ -170.35.

[0326] Example 111: (R)-2-fluoro-3-hydroxy-2-phenyl-1-(5-(pyridin-2-ylsulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)propan-1-one-3,3-d2

[0327] First Step: Synthesis of ethyl 2-fluoro-3-hydroxy-2-phenylpropanoate-3,3-d2

[0328] To ethyl 2-fluoro-2-phenylacetate (0.5 g, 2.74 mmol) in DMF (10 mL) was added paraformaldehyde-D2 (0.26 g, 2.74 mmol) and sodium ethoxide (0.14 g, 0.41 mmol) at room temperature and stirred at room temperature. After the reaction was quenched with water (20 mL), extracted with ethyl acetate (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 4 / 1) to give the title compound (0.48 g, 81.64%) as a light yellow oil.

[0329] 1 H NMR (400 MHz, CDCl3) δ 7.54 - 7.52 (m, 2H), 7.41 (q, J = 5.8 Hz, 3H), 4.33 - 4.28 (m, 2H), 1.31 (t, J = 7.1 Hz, 3H). 19 F NMR (376 MHz, CDCl3) δ -171.85 (s).

[0330] Second Step: Synthesis of 2-fluoro-3-hydroxy-2-phenylpropanoic acid-3,3-d2

[0331] To a solution of ethyl 2-fluoro-3-hydroxy-2-phenylpropanoate-3,3-d2 (0.5 g, 2.33 mmol) in water (5 mL) and methanol (10 mL) was added lithium hydroxide monohydrate (0.20 g, 4.66 mmol) at room temperature and stirred at room temperature until the reaction was completed. Concentrated under reduced pressure, the residue was adjusted to pH = 1 with dilute hydrochloric acid (1 M), stirred for 10 minutes, and then concentrated under reduced pressure. The most of the organic phase was removed, and then the system was diluted with ethyl acetate (20 mL) and washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give the target compound (0.4 g, 92.05%) as a white solid.

[0332] 1H NMR (400 MHz, DMSO-d6) δ 7.47 (d, J = 7.1 Hz, 2H), 7.44 - 7.34 (m, 3H).

[0333] 19 F NMR (376 MHz, DMSO-d6) δ -167.09 (s).

[0334] Third Step: Synthesis of (R)-2-fluoro-3-hydroxy-2-phenylpropanoic acid-3,3-d2

[0335] To a solution of 2-fluoro-3-hydroxy-2-phenylpropanoic acid-3,3-d2 (1.8 g, 9.67 mmol) in ethanol (18 mL), (1R,2R)-2-amino-1-(4-nitrophenyl)propane-1,3-diol (0.98 g, 4.64 mmol) was added, heated to reflux and stirred. After the reaction was completely dissolved, it was naturally cooled to room temperature, cooled with an ice bath, then filtered, the filter cake was washed with cooled ethanol, then the filter cake was collected, ethanol (18 mL) was added, heated to reflux for 3 hours, naturally cooled to room temperature, cooled with an ice bath, filtered, the filter cake was washed with cold ethanol, dried under reduced pressure, the solid was diluted with water, then slowly adjusted to pH = 1 with 1M dilute hydrochloric acid, continued to stir for 10 minutes, extracted with ethyl acetate (30 mL x 5), the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated to give the target compound (0.65 g, 36.11%) as a white solid.

[0336] 1 H NMR (599 MHz, DMSO-d6) δ 7.48 (d, J = 7.4 Hz, 2H), 7.41 - 7.37 (m, 3H).

[0337] Fourth Step: Synthesis of (R)-2-fluoro-3-hydroxy-2-phenyl-1-(5-(pyridin-2-ylsulfonyl)- 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)propan-1-one-3,3-d2

[0338] To a solution of 2-((lH-pyrazol-4-yl)sulfonyl)-l,2,3,4,5,6-hexahydropyrrolo[3,4- c]pyrrole dihydrochloride (0.36 g, 1.12 mmol), (R)-2-fluoro-3-hydroxy-2-phenylpropanoic acid- 3,3-d2 (0.2 g, 1.07 mmol) in DMF (10 mL) was added HOBt (0.22 g, 1.60 mmol) and DIPEA (0.71 mL, 4.28 mmol) at room temperature, stirred for 5 min, then EDCI (0.27 g, 1.39 mmol) was added, after addition, stirred at room temperature overnight. The reaction solution was quenched with water (10 mL), extracted with ethyl acetate (10 mL x 2), the organic phase was washed with saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1 / 1) to give the title compound (0.23 g, 51.04%) as a white solid. MS (ESI, pos.ion) m / z: 420.1 [M+H] + ;

[0339] 1 H NMR (599 MHz, DMSO-d6) d 8.71 (d, J = 4.6 Hz, 1H), 8.10 - 8.09 (m, 1H), 7.95 (t, J = 7.8 Hz, 1H), 7.68 (dd, J = 7.6, 4.7 Hz, 1H), 7.45 - 7.35 (m, 5H), 5.26 (s, 1H), 4.44 (d, J = 15.4 Hz, 1H), 4.29 - 4.12 (m, 5H), 4.09 (d, J = 15.6 Hz, 1H), 3.97 (d, J = 14.4 Hz, 1H).

[0340] Example 112: (R)-2-fluoro-3-hydroxy-2-(phenyl-d5)-l-(5-(pyridin-2-ylsulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)propan-l-one-3,3-d2

[0341] First Step: Synthesis of 2-fluoro-3-hydroxy-2-(phenyl-d5)ethyl propanoate-3,3-d2

[0342] Ethyl 2-fluoro-2-(phenyl-d5)acetate (2.0 g, 10.68 mmol) was dissolved in DMF (15 mL), deuterated paraformaldehyde (1.03 g, 10.68 mmol), sodium ethoxide (0.84 mL, 2.14 mmol) were added and the reaction was stirred at room temperature. After quenching with water (40 mL), the reaction was extracted with ethyl acetate (3 x 30 mL), the organic phases were combined, washed with saturated brine (30 mL) and dried over anhydrous sodium sulfate. After filtration, the solvent was removed under reduced pressure and the residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 6 / 1) to give the title compound as a yellowish oil (1.40 g, 59.77%).

[0343] MS (ESI, pos.ion) m / z: 220.0 [M+H] + ; 1 H NMR (400 MHz, CDC13) δ 4.35 - 4.28 (m, 2H), 1.32 (t, J = 7.1 Hz, 3H). 19 F NMR (376 MHz, CDC13) δ -172.04.

[0344] Second step: synthesis of ethyl 2-fluoro-3-hydroxy-2-(phenyl-d5)propanoate-3,3-d2

[0345] Ethyl 2-fluoro-3-hydroxy-2-(phenyl-d5)propanoate-3,3-d2 (1.40 g, 6.39 mmol) was dissolved in tetrahydrofuran (20 mL), lithium hydroxide monohydrate (0.38 g, 15.97 mmol) dissolved in water (4 mL) was added and the reaction was stirred at room temperature until completion. The organic solvent was removed by rotary evaporation and the residue was dissolved in water (10 mL) and the pH was adjusted to 1 with dilute hydrochloric acid (1 M). The reaction was extracted with ethyl acetate (5 x 25 mL), washed with saturated brine (20 mL) and dried over anhydrous sodium sulfate. The title compound was obtained as a white solid after concentration under reduced pressure (0.58 g, 47.51%). MS (ESI, neg.ion) m / z: 190.1 [M-H] - ;

[0346] Third step: synthesis of (R)-2-fluoro-3-hydroxy-2-(phenyl-d5)propanoic acid-3,3-d2

[0347] To a solution of 2-fluoro-3-hydroxy-2-(phenyl-d5)propanoic acid-3,3-d2 (0.56 g, 2.93 mmol) in ethanol (10 mL), (1R,2R)-2-amino-1-(4-nitrophenyl)propane-1,3-diol (0.28 g, 1.32 mmol) was added and stirred at reflux. After the reaction was completely dissolved, it was naturally cooled to room temperature, further cooled with an ice bath, then filtered, the filter cake was washed with cooled ethanol, then the filter cake was collected, ethanol (10 mL) was added, heated to reflux for 3 hours, naturally cooled to room temperature, cooled with an ice bath, filtered, the filter cake was washed with cold ethanol, and dried under reduced pressure. The solid was diluted with water, then slowly adjusted to pH = 1 with 1M dilute hydrochloric acid, stirred for 10 minutes, extracted with ethyl acetate (25 mL x 5), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the white solid target compound (0.22 g, 78.57%).

[0348] MS (ESI, neg. ion) m / z: 190.1 [M-H] - .

[0349] Fourth step: synthesis of (R)-2-fluoro-3-hydroxy-2-(phenyl-d5)-1-(5-(pyridin-2- ylsulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)propan-1-one-3,3-d2

[0350] 2-(Pyridin-2-sulfonyl)-1,2,3,4,5,6-hexahydropyrrolo[3,4-c]pyrrole dihydrochloride (0.33 g, 1.02 mmol) was dissolved in tetrahydrofuran (10 mL), DIPEA (0.51 mL, 3.06 mmol), HOBt (0.21 g, 1.53 mmol), EDCI (0.29 g, 1.53 mmol), (R)-2-fluoro-3-hydroxy-2-(phenyl-d5)propanoic acid-3,3-d2 (0.20 g, 1.02 mmol) were added, and stirred at room temperature under nitrogen protection until the reaction was complete. After the reaction was completed, the reaction was quenched by adding an appropriate amount of water, extracted with ethyl acetate (3 x 25 mL), and the organic layer was taken, washed with saturated brine (25 mL), and dried over anhydrous sodium sulfate. After filtration, the residue was concentrated under reduced pressure, and the white solid title compound was obtained by silica gel column chromatography separation and purification (petroleum ether / ethyl acetate (v / v) = 1 / 3) (0.12 g, 27.28%). MS (ESI, pos. ion) m / z: 425.1 [M+H] + ; 1H NMR (599 MHz, DMSO-d6) δ 8.71 (dd, J = 4.8, 1.6 Hz, 1H), 8.11 - 8.08 (m, 1H), 7.94 (d, J = 7.8 Hz, 1H), 7.68 (dd, J = 7.7, 4.6 Hz, 1H), 5.27 (s, 1H), 4.45 (dd, J = 14.6, 4.9 Hz, 1H), 4.25 - 4.13 (m, 5H), 4.11 - 4.09 (m, 1H), 3.99-3.96 (m, 1H).

[0351] Example 113: (R)-2-fluoro-3-hydroxy-2-phenyl-1-(5-((pyridin-2-yl-d4)sulfonyl)- 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)propan-1-one

[0352] First Step: Synthesis of pyridine-2-sulfonyl chloride-d4

[0353] Under nitrogen protection, slowly drop butylmagnesium chloride (0.36 g, 2.62 mmol) into n-butyllithium (0.16 g, 2.47 mmol) at -10 °C, then slowly drop 2-bromopyridine-d4 (0.8 g, 4.94 mmol) in THF (30 mL) solution, continue to react for 1 h, then drop sulfuryl chloride (6.67 g, 49.40 mmol) continue to react. TLC monitor the reaction is complete, stop the reaction, remove the organic solvent under reduced pressure, extract with ethyl acetate (30 mL x 3), separate, dry over anhydrous sodium sulfate, filter, concentrate under reduced pressure, the residue is separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10 / 1), to get the title compound (0.12 g, 13.38%) as orange oil.

[0354] Second Step: Synthesis of tert-butyl 5-((pyridin-2-yl-d4)sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate

[0355] In the solution of pyridine-2-sulfonyl chloride-d4 (0.12 g, 0.66 mmol) in dichloromethane (10 mL), add tert-butyl 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-2(1H)- carboxylate p-toluenesulfonic acid salt (0.25 g, 0.66 mmol) and triethylamine (0.20 g, 1.98 mmol), continue to react at room temperature. Concentrate under reduced pressure, the residue is separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 5 / 1), to get the title compound (0.13 g, 55.36%) as white solid.

[0356] Step 3: Synthesis of 2-((pyridin-2-yl-d4)sulfonyl)-l,2,3,4,5,6-hexahydropyrrolo[3,4- c]pyrrole dihydrochloride

[0357] To a solution of tert-butyl 5-((pyridin-2-yl-d4)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4- c]pyrrole-2(lH)-carboxylate (0.13 g, 0.37 mmol) in ethyl acetate (20 mL) was added hydrochloric acid (0.13 g, 3.7 mmol) in ethyl acetate solution, and the reaction was carried out at room temperature. TLC monitoring, concentrated under reduced pressure, to obtain the target compound (0.11 g, 98.39%) as a red-brown solid.

[0358] Step 4: Synthesis of (R)-2-fluoro-3-hydroxy-2-phenyl-l-(5-((pyridin-2-yl-d4)sulfonyl)- 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)propan-l-one

[0359] To a solution of 2-((pyridin-2-yl-d4)sulfonyl)-l,2,3,4,5,6-hexahydropyrrolo[3,4- c]pyrrole dihydrochloride (120 mg, 0.41 mmol) in DMF (10 mL) was added (R)-2-fluoro-3- hydroxy-2-phenylpropanoic acid (60 mg, 0.33 mmol), EDCI (76 mg, 0.40 mmol) and HOBt (53.51 mg, 0.40 mmol) successively, stirred and dissolved, and finally DIPEA (170.60 mg, 1.32 mmol) was added, and the reaction was continued at room temperature. TLC monitoring showed that the reaction was complete, the reaction was stopped, water (20 ml) was added, extracted with ethyl acetate (10 mL x 3), separated, the organic phase was washed with saturated sodium chloride solution (20 mL x 2), separated, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 2 / 1) to obtain the title compound (25 mg, 18.21%) as a yellow solid. MS (ESI, pos.ion) m / z: 422.15 [M+H] + ; 1 H NMR (599 MHz, DMSO-d6) δ 7.51 - 7.33 (m, 5H), 5.29 (t, J = 6.1 Hz, 1H), 4.43 (d, J = 15.4 Hz, 1H), 4.27 - 4.06 (m, 7H), 3.96 (d, J = 15.3 Hz, 1H), 3.73 - 3.62 (m, 1H).

[0360] Example 114: (R)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-yl-2,2,3,3-d4) sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3-hydroxy-2- phenylpropan-1-one

[0361] First Step: Synthesis of 2-((5-bromo-2-chloropyridin-3-yl)oxy)ethane-1,1,2,2-d4-1-ol

[0362] Triphenylphosphine (5.66 g, 21.59 mmol) was dissolved in tetrahydrofuran (40 ml), DIAD (4.25 ml, 21.59 mmol) was added dropwise at 0 °C, after the addition was completed, stirred at room temperature for 30 min, then added 5-bromo-2-chloropyridin-3-ol (3.0 g, 14.39 mmol), ethane-d4-1,2-diol (0.88 mL, 14.39 mmol), and the reaction was completed after stirring at room temperature. The reaction solution was poured into water (20 mL), extracted with ethyl acetate (3 x 20 mL), the organic layers were combined and washed with saturated brine (20 mL) and dried over anhydrous sodium sulfate. Filtration, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 4 / 1) to give the title compound (2.47 g, 66.90%) as a white solid.

[0363] MS (ESI, pos.ion) m / z: 258.0 [M+H] + .

[0364] Second Step: Synthesis of 7-bromo-2,3-dihydro-[1,4]dioxino[2,3-b]pyridine-2,2,3,3-d4

[0365] 2-((5-bromo-2-chloropyridin-3-yl)oxy)ethane-1,1,2,2-d4-1-ol (2.45 g, 9.55 mmol) was dissolved in toluene (40 ml), potassium hydroxide (0.80 g, 14.33 mmol), 18-crown-6 ether (0.33 g, 1.24 mmol) was added, and the reaction was completed after stirring at 110 °C. Diluted with water (35 mL), extracted with ethyl acetate (3 x 35 mL), and the organic layer was obtained, washed with saturated brine (35 mL) and dried over anhydrous sodium sulfate. Filtration, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 4 / 1) to give the title compound (0.79 g, 37.59%) as a white solid.

[0366] MS (ESI, pos.ion) m / z: 221.0 [M+H] + .

[0367] Step 3: Synthesis of 7-(benzylthio)-2,3-dihydro-[l,4]dioxino[2,3-b]pyridine-2,2,3,3-d4

[0368] 7-bromo-2,3-dihydro-[l,4]dioxino[2,3-b]pyridine-2,2,3,3-d4 (0.85 g, 3.86 mmol) was dissolved in 1,4-dioxane (15 mL), benzyl mercaptan (0.45 mL, 3.86 mmol), DIPEA (1.91 mL, 11.58 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.45 g, 0.77 mmol), tris(dibenzylideneacetone)dipalladium (0.35 g, 0.39 mmol) were added, and the reaction was stirred at 100 °C under nitrogen until the reaction was completed. After the reaction was completed, the reaction solution was cooled to room temperature, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 4 / 1) to obtain the title compound (1.0 g, 98.31%) as a yellow oil. MS (ESI, pos.ion) m / z: 264.1 [M+H] + .

[0369] Step 4: Synthesis of 2,3-dihydro-[l,4]dioxino[2,3-b]pyridine-7-sulfonyl chloride-2,2,3,3-d4

[0370] 7-(benzylthio)-2,3-dihydro-[l,4]dioxino[2,3-b]pyridine-2,2,3,3-d4 (1.0 g, 3.80 mmol) was dissolved in acetonitrile (10 mL), NCS (1.78 g, 13.30 mmol), acetic acid (1.0 mL), and water (0.5 mL) were added, and the reaction was stirred at room temperature until the reaction was completed. The reaction was quenched by adding saturated sodium bisulfite aqueous solution (30 mL), extracted with dichloromethane (3 x 25 mL), and the organic layer was obtained, washed with saturated brine (25 mL), and dried over anhydrous sodium sulfate. Concentration under reduced pressure gave the title compound (0.90 g, 98.89%) as a light yellow oil. MS (ESI, pos.ion) m / z: 240.0 [M+H] + .

[0371] Step 5: Synthesis of tert-butyl 5-((2,3-dihydro-[l,4]dioxino[2,3-b]pyridin-7-yl-2,2,3,3-d4) sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-2(lH)-carboxylate

[0372] tert-Butyl 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-2(lH)-carboxylate p-toluenesulfonic acid salt (1.40 g, 3.66 mmol) was dissolved in dichloromethane (20 mL), triethylamine (1.27 mL, 9.15 mmol), 2,3-dihydro-[l,4]dioxino[2,3-b]pyridine-6-sulfonyl chloride (0.88 g, 3.66 mmol) were added, and stirring was performed at room temperature until the reaction was completed. Water (20 mL) was added to quench the reaction, and extraction was performed with dichloromethane (3 x 30 mL). The combined organic layers were washed with saturated brine (30 mL) and dried over anhydrous sodium sulfate. Concentration under reduced pressure was performed, and the residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 1 / 1) to obtain the title compound as a light yellow solid (0.65 g, 42.95%). + .

[0373] Step 6: Synthesis of 7-((3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)sulfonyl)- 2,3-dihydro-[l,4]dioxino[2,3-b]pyridine-2,2,3,3-d4 hydrochloride

[0374] tert-Butyl 5-((2,3-dihydro-[l,4]dioxino[2,3-b]pyridin-7-yl-2,2,3,3-d4)sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrole-2(lH)-carboxylate (0.63 g, 1.52 mmol) was dissolved in ethyl acetate (10 mL), and a solution of HC1 (2.30 mL, 9.2 mmol, 4.0 M) in 1,4-dioxane was added. Stirring was performed at room temperature. After the reaction was completed, rotary evaporation was performed under reduced pressure to obtain the target compound as a light pink solid (0.49 g, 91.92%).

[0375] MS (ESI, pos.ion) m / z: 314.1 [M+H] + .

[0376] Step 7: Synthesis of (R)-l-(5-((2,3-dihydro-[l,4]dioxino[2,3-b]pyridin-7-yl-2,2,3,3-d4)sulfonyl)- 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)-2-fluoro-3-hydroxy-2-phenylpropan-l-one

[0377] To a solution of 7-((3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)sulfonyl)-2,3- dihydro-[l,4]dioxino[2,3-b]pyridine-2,2,3,3-d4 hydrochloride (0.49 g, 1.40 mmol) in tetrahydrofuran (10 mL) was added DIPEA (0.69 mL, 4.20 mmol), HOBt (0.28 g, 2.10 mmol), EDCI (0.40 g, 2.10 mmol), (R)-2-fluoro-3-hydroxy-2-(4-chloromethylphenyl)propanoic acid (0.26 g, 1.40 mmol) under nitrogen protection. The mixture was stirred at room temperature until the reaction was completed. After the reaction was completed, the reaction was quenched by water (25 mL), extracted with ethyl acetate (3 x 25 mL), the organic layer was collected, washed with saturated brine (25 mL), and dried over anhydrous sodium sulfate. The residue was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1 / 4) to give the title compound (0.42 g, 62.54%) as a white solid. MS (ESI, pos.ion) m / z: 480.1 [M+H] + ;

[0378] 1 H NMR (599 MHz, DMSO-d6) d 8.14 (d, J = 2.2 Hz, 1H), 7.61 (d, J = 2.2 Hz, 1H), 7.43 - 7.35 (m, 5H), 5.30 (t, J = 6.1 Hz, 1H), 4.46 - 4.40 (m, 1H), 4.17 - 4.01 (m, 7H), 3.97 - 3.92 (m, 1H), 3.69 (ddd, J = 20.1, 12.3, 5.9 Hz, 1H).

[0379] Example 115: (R)-2-fluoro-3-hydroxy-l-(5-((5-(methoxy-d3)pyridin-2-yl)sulfonyl)- 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)-2-phenylpropan-l-one

[0380] First Step: Synthesis of 2-bromo-5-(methoxy-d3)pyridine

[0381] To a solution of 6-bromopyridin-3-ol (3 g, 17.24 mmol) in DMF (20 mL), cesium carbonate (8.43 g, 25.86 mmol) was added, the reaction was stirred at room temperature for 15 min, then deuterated methyl iodide (2.70 g, 18.62 mmol) was added, the reaction was continued to stir at 45 °C until the reaction was completed. Then quenched with water (50 mL), extracted with ethyl acetate (25 mL x 3), separated, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 4 / 1) to give the title compound (3.12 g, 94.72%) as a yellow oil.

[0382] MS (ESI, pos.ion) m / z: 191.10 [M+H] + .

[0383] Second step: synthesis of 2-(benzylthio)-5-(methoxy-d3)pyridine

[0384] To a solution of 2-bromo-5-(methoxy-d3)pyridine (3.0 g, 15.70 mmol) in 1,4-dioxane (30 mL), Pd2(dba)3(0.72 g, 0.79 mmol) was added and stirred to dissolve, then Xantphos (0.91 g, 1.57 mmol) and DIPEA (4.06 g, 31.4 mmol) were added, replaced with nitrogen, and finally benzyl mercaptan (1.95 g, 15.7 mmol) was added and the reaction was heated to 100 °C. TLC showed that the reaction was complete, and after natural cooling to room temperature, the organic solvent was removed by concentration under reduced pressure, diluted with water (45 mL), extracted with ethyl acetate (20 mL x 3), separated, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10 / 1) to give the title compound (3.2 g, 86.96%) as a yellow oil.

[0385] MS (ESI, pos.ion) m / z: 235.1 [M+H] + ;

[0386] 1 H NMR (599 MHz, CDCl3) δ 8.20 (d, J = 2.9 Hz, 1H), 7.36 (d, J = 7.3 Hz, 2H), 7.26 (dd, J = 13.9, 6.2 Hz, 2H), 7.21 (t, J = 7.3 Hz, 1H), 7.09 (d, J = 8.7 Hz, 1H), 7.05 (dd, J = 8.7, 2.9 Hz, 1H), 4.37 (s, 2H).

[0387] Third step: synthesis of 5-(methoxy-d3)pyridine-2-sulfonyl chloride

[0388] To a solution of 2-(benzylsulfanyl)-5-(methoxy-d3)pyridine (0.4 g, 1.71 mmol) in dichloromethane (10 mL), NCS (1.14 g, 8.55 mmol) and benzyl alcohol (1.29 g, 11.97 mmol) were added, and the reaction was continued at room temperature. TLC monitoring showed that the reaction was complete, and the reaction was stopped. The residue was concentrated under reduced pressure and purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 5 / 1) to give the title compound (0.19 g, 52.84%) as a transparent oil.

[0389] Fourth step: synthesis of tert-butyl 5-((5-(methoxy-d3)pyridin-2-yl)sulfonyl)- 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-2(lH)-carboxylate

[0390] To a solution of 5-(methoxy-d3)pyridine-2-sulfonyl chloride (0.8 g, 3.80 mmol) in dichloromethane (30 mL), tert-butyl 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-2(lH)- carboxylate p-toluenesulfonate (1.45 g, 3.8 mmol) was added and stirred to dissolve, then triethylamine (1.15 g, 11.40 mmol) was added, and the reaction was continued at room temperature. TLC monitoring showed that the reaction was complete, and the reaction was stopped. The residue was concentrated under reduced pressure and purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 5 / 1) to give the title compound (0.95 g, 65.06%) as a white solid.

[0391] 1 H NMR (599 MHz, CDC13) δ 8.33 (d, J = 2.7 Hz, 1H), 7.92 (d, J = 8.6 Hz, 1H), 7.29 (dd, J = 8.7, 2.8 Hz, 1H), 4.28 (d, J = 16.2 Hz, 4H), 4.03 (d, J = 7.7 Hz, 4H), 1.45 (s, 9H).

[0392] Fifth step: synthesis of 2-((5-(methoxy-d3)pyridin-2-yl)sulfonyl)-l,2,3,4,5,6- hexahydropyrrolo[3,4-c]pyrrole hydrochloride

[0393] To a solution of tert-butyl 5-((5-(methoxy-d3)pyridin-2-yl)sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrole-2(lH)-carboxylate (0.95 g, 2.47 mmol) in ethyl acetate (20 mL) was added HCl (6.1 mL, 24.70 mmol, 4 M) in ethyl acetate and the reaction was allowed to proceed at room temperature. The reaction was monitored by TLC and upon completion, the reaction was stopped and concentrated under reduced pressure to obtain the title compound as a red-brown solid (0.80 g, 99%).

[0394] Sixth Step: Synthesis of (R)-2-fluoro-3-hydroxy-l-(5-((5-(methoxy-d3)pyridin-2- yl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)-2-phenylpropan-l-one

[0395] To a solution of 2-((5-(methoxy-d3)pyridin-2-yl)sulfonyl)-l,2,3,4,5,6- hexahydropyrrolo[3,4-c]pyrrole hydrochloride (0.84 g, 2.60 mmol) in DMF (20 mL) was added (R)-2-fluoro-3-hydroxy-2-(4-chloromethylphenyl)propanoic acid (0.40 g, 2.17 mmol), EDCI (0.50 g, 2.60 mmol) and HOBt (0.35 g, 2.60 mmol) successively, stirred to dissolve and finally DIPEA (1.12 g, 8.68 mmol) was added and the reaction was allowed to proceed at room temperature. The reaction was monitored by TLC and upon completion, the reaction was stopped, water (60 mL) was added and the reaction was extracted with ethyl acetate (30 mL x 3), the organic layers were combined, washed with saturated sodium chloride solution (30 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 5 / 1) to obtain the title compound as a white solid (0.53 g, 54.16%).

[0396] MS (ESI, pos.ion) m / z: 451.1 [M+H] + ;

[0397] 1 H NMR (599 MHz, DMSO-d6) δ 8.39 (d, J = 2.9 Hz, 1H), 7.89 (d, J = 8.7 Hz, 1H), 7.58 (dd, J = 8.7, 2.9 Hz, 1H), 7.44 - 7.34 (m, 5H), 5.30 (t, J = 6.1 Hz, 1H), 4.43 (d, J = 15.5 Hz, 1H), 4.23 - 4.05 (m, 7H), 3.95 (d, J = 15.0 Hz, 1H), 3.68 (ddd, J = 19.9, 12.3, 6.0 Hz, 1H).

[0398] Example 116 (R)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl)sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3-hydroxy-2-phenylpropan-1-one

[0399] First Step: Synthesis of 6-bromo-2-chloropyridin-3-ol

[0400] 2-chloropyridin-3-ol (3.0 g, 23.16 mmol) was dissolved in DMF (20 mL), a solution of NBS (3.71 g, 20.84 mmol) in DMF (20 mL) was added dropwise at -10 °C, after the addition was completed, the reaction was continued to end. Add saturated sodium bisulfite (20 mL) to quench the reaction, extract with dichloromethane (3 x 35 mL), wash with saturated brine (30 mL), dry over anhydrous sodium sulfate. Filter, concentrate under reduced pressure, and purify the residue by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10 / 1) to give the title compound as a white solid (0.95 g, 19.68%).

[0401] MS (ESI, neg. ion) m / z: 207.8 [M-H] - .

[0402] Second Step: Synthesis of 2-((6-bromo-2-chloropyridin-3-yl)oxy)ethan-1-ol

[0403] 6-bromo-2-chloropyridin-3-ol (0.53 g, 2.54 mmol) was dissolved in DMF (5 mL), 2-bromoethan-1-ol (0.54 mL, 7.62 mmol), potassium carbonate (0.88 mL, 6.35 mmol) were added, and stirred at 120 °C until the reaction was completed. Dilute with water (25 mL), extract with ethyl acetate (3 x 25 mL), take the organic layer, wash with saturated brine (25 mL), dry over anhydrous sodium sulfate. Concentrate under reduced pressure, and purify the residue by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 2 / 1) to give the title compound as a light yellow oil (0.51 g, 79.44%).

[0404] MS (ESI, pos. ion) m / z: 253.9 [M+H] + .

[0405] Third Step: Synthesis of 6-bromo-2,3-dihydro-[1,4]dioxino[2,3-b]pyridine

[0406] Dissolve 2-((6-bromo-2-chloropyridin-3-yl)oxy)ethan-1-ol (0.49 g, 1.94 mmol) in toluene (10 mL), add potassium hydroxide (0.16 g, 2.91 mmol), 18-crown-6 (0.07 g, 0.25 mmol), stir the reaction at 110 °C until the reaction is completed. Dilute with water (25 mL), extract with ethyl acetate (3 x 25 mL), take the organic layer, wash with saturated brine (25 mL), dry over anhydrous sodium sulfate. Filter, concentrate under reduced pressure, purify the residue by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 6 / 1), to give the title compound (0.27 g, 64.4%) as a white solid.

[0407] MS (ESI, pos.ion) m / z: 217.9 [M+H] + .

[0408] Fourth step: synthesis of 6-(benzylthio)-2,3-dihydro-[1,4]dioxino[2,3-b]pyridine

[0409] Dissolve 6-bromo-2,3-dihydro-[1,4]dioxino[2,3-b]pyridine (0.73 g, 3.38 mmol) in 1,4-dioxane (10 mL), add benzyl mercaptan (0.60 mL, 5.07 mmol), DIPEA (1.68 mL, 10.14 mmol), Xant-Phos (0.39 g, 0.68 mmol), tris(dibenzylideneacetone)dipalladium (0.31 g, 0.34 mmol), protect with nitrogen, stir at 100 °C until the reaction is completed. Cool the reaction to room temperature, concentrate under reduced pressure, purify the residue by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10 / 1), to give the title compound (0.80 g, 91.29%) as a yellow oil.

[0410] MS (ESI, pos.ion) m / z: 260.0 [M+H] + .

[0411] Fifth step: synthesis of 2,3-dihydro-[1,4]dioxino[2,3-b]pyridine-6-sulfonyl chloride

[0412] To a solution of 6-(benzylsulfanyl)-2,3-dihydro-[l,4]dioxino[2,3-b]pyridine (0.80 g, 3.08 mmol) in acetonitrile (10 mL) was added NCS (1.44 g, 10.78 mmol), acetic acid (1.0 mL), water (0.5 mL) and stirred at room temperature until the reaction was completed. The reaction was quenched by adding saturated sodium bisulfite (20 mL) aqueous solution and extracted with dichloromethane (3 x 20 mL). The organic layer was collected, washed with saturated brine (20 mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, the residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 6 / 1) to give the title compound (0.66 g, 90.79%) as a light yellow oil. MS (ESI, pos.ion) m / z: 235.9 [M+H] + .

[0413] Step 6: Synthesis of tert-butyl 5-((2,3-dihydro-[l,4]dioxino[2,3-b]pyridin-6- yl)sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-2(lH)-carboxylate

[0414] To a solution of tert-butyl 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-2(lH)- carboxylate p-toluenesulfonate (1.0 g, 2.61 mmol) in dichloromethane (15 ml) was added triethylamine (0.91 mL, 6.52 mmol), 2,3-dihydro-[l,4]dioxino[2,3-b]pyridine-6-sulfonyl chloride (0.62 g, 2.61 mmol) and stirred at room temperature until the reaction was completed. The reaction was quenched by adding water (20 mL) and extracted with dichloromethane (3 x 25 mL). The organic layer was collected, washed with saturated brine (25 mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, the residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 1 / 1) to give the title compound (0.81 g, 75.66%) as a light yellow solid. MS (ESI, pos.ion) m / z: 410.0 [M+H] + .

[0415] Step 7: Synthesis of 6-((3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)sulfonyl)- 2,3-dihydro-[l,4]dioxino[2,3-b]pyridine hydrochloride

[0416] Tert-butyl 5-((2,3-dihydro-[l,4]dioxino[2,3-b]pyridin-6-yl)sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrole-2(lH)-carboxylate (0.81 g, 1.98 mmol) was dissolved in HCl in 1,4-dioxane (1.5 mL, 5.94 mmol, 4 M) and stirred at room temperature until the reaction was complete. Concentrated under reduced pressure to obtain the title compound (0.51 g, 67.43%) as a light pink solid. MS (ESI, pos.ion) m / z: 310.0 [M+H] + .

[0417] Step 8: (R)-l-(5-((2,3-dihydro-[l,4]dioxino[2,3-b]pyridin-6-yl)sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)-2-fluoro-3-hydroxy-2-phenylpropan-l-one

[0418] 6-((3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)sulfonyl)-2,3-dihydro- [l,4]dioxino[2,3-b]pyridine hydrochloride (0.51 g, 1.65 mmol) was dissolved in tetrahydrofuran (10 mL), DIPEA (0.82 ml, 4.95 mmol), HOBt (0.33 g, 2.47 mmol), EDCI (0.47 g, 2.47 mmol), (R)-2-fluoro-3-hydroxy-2-(4-chloromethylphenyl)propanoic acid (0.30 g, 1.65 mmol) were added, and stirred at room temperature under nitrogen until the reaction was complete. After the reaction was completed, water (20 mL) was added to quench, and extracted with ethyl acetate (3 x 25 mL), and the organic layer was washed with saturated brine (30 mL) and dried over anhydrous sodium sulfate. Filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1 / 4) to obtain the title compound (0.43 g, 54.85%) as a white solid. MS (ESI, pos.ion) m / z: 476.0 [M+H] + ;

[0419] 1 H NMR (400 MHz, DMSO-d6) d 7.46 (s, 2H), 7.42 - 7.33 (m, 5H), 5.31 (t, J = 6.1 Hz, 1H), 4.48 - 4.39 (m, 3H), 4.35 - 4.28 (m, 2H), 4.17-4.05 (m, 7H), 3.99 - 3.92 (m, 1H), 3.68 (ddd, J = 19.2, 12.4, 6.1 Hz, 1H).

[0420] 19F NMR (376 MHz, DMSO-d6) δ -170.39.

[0421] Example 117 (R)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl) sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3-hydroxy-2-(phenyl-d5)propan-1-one

[0422] To the reaction flask was added (R)-2-fluoro-3-hydroxy-2-(phenyl-d5)propanoic acid (0.54 g, 2.85 mmol), 6-((3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)sulfonyl)-2,3-dihydro- [1,4]dioxino[2,3-b]pyridine hydrochloride (1.14 g, 2.99 mmol), HOBt (0.58 g, 4.28 mmol), DMF (10 mL) and DIPEA (1.47 g, 11.4 mmol) successively, after stirring for 5 minutes, EDCI (0.71 g, 3.71 mmol) was added, the reaction was stirred at room temperature for 5 hours. After the reaction was completed, water (60 mL) was added for dilution, extracted with ethyl acetate (60 mL x 2), the organic phase was collected, washed with water (100 mL x 2), saturated brine (100 mL) successively, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 3 / 7) to give the title compound (0.73 g, 53.23%) as a white solid. MS (ESI, pos.ion) m / z: 481.2 [M+H] + ;

[0423] 1 H NMR (600 MHz, DMSO-d6) δ 7.50 - 7.46 (m, 2H), 5.31 (t, J = 6.1 Hz, 1H), 4.49 - 4.43 (m, 3H), 4.36 - 4.31 (m, 2H), 4.21 - 4.06 (m, 7H), 4.01 - 3.95 (m, 1H), 3.69 (ddd, J = 20.0, 12.3, 5.9 Hz, 1H).

[0424] Example 118 (R)-1-(5-((2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-6-yl-2,2,3,3-d4) sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2-fluoro-3-hydroxy-2-phenylpropan-1- one

[0425] Step 1: Synthesis of 2-((6-bromo-2-chloropyridin-3-yl)oxy)ethan-1,1,2,2-d4-1-ol

[0426] Triphenylphosphine (5.63 g, 21.45 mmol) was dissolved in tetrahydrofuran (30 mL), DIAD (4.34 g, 21.45 mmol) was added dropwise slowly under stirring at 0 °C, after the addition was completed, 6-bromo-2-chloropyridin-3-ol (2.98 g, 14.30 mmol) and deuterated ethylene glycol (0.95 g, 14.3 mmol) were added after stirring at room temperature for 30 min, and the reaction was carried out at room temperature overnight. After the reaction was completed, the reaction liquid was poured into water, extracted with ethyl acetate (35 mL x 2), the organic layer was taken, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 3 / 2) to obtain the title compound (3.5 g, 95.44%) as a white solid.

[0427] Step 2: Synthesis of 6-bromo-2,3-dihydro-[1,4]dioxino[2,3-b]pyridine-2,2,3,3-d4

[0428] Into the reaction bottle, 2-((6-bromo-2-chloropyridin-3-yl)oxy)ethan-1,1,2,2-d4-1-ol (3.5 g, 13.64 mmol) was dissolved in toluene (50 mL), potassium hydroxide (1.15 g, 20.46 mmol) and 18-crown-6 ether (0.47 g, 1.747 mmol) were added, and the reaction was carried out at 110 °C for 1.5 hours. After cooling to room temperature, water (30 mL) was added for dilution, extracted with ethyl acetate (35 mL x 3), the organic layer was taken, washed with saturated brine (35 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10 / 1) to obtain the title compound (0.63 g, 20.98%) as a white solid. MS (ESI, pos.ion) m / z: 221.1 [M+H] + .

[0429] Step 3: Synthesis of 6-(benzylthio)-2,3-dihydro-[1,4]dioxino[2,3-b]pyridine-2,2,3,3-d4

[0430] To a reaction flask was added 6-bromo-2,3-dihydro-[l,4]dioxino[2,3-b]pyridine- 2,2,3,3-d4 (0.63 g, 2.86 mmol), tris(dibenzylideneacetone)dipalladium (0.33 g, 0.57 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.26 g, 0.29 mmol), DIPEA (1.11 g, 8.58 mmol) and 1,4-dioxane (15 mL), after replaced with nitrogen, added benzyl mercaptan (0.36 g, 2.86 mmol), replaced with nitrogen again, then heated to 100 °C and stirred overnight. Concentrated under reduced pressure, the residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 10 / 1) to give the title compound (0.66 g, 87.54%) as yellow oil. MS (ESI, pos.ion) m / z: 264.1 [M+H] + .

[0431] Fourth step: synthesis of 2,3-dihydro-[l,4]dioxino[2,3-b]pyridine-6-sulfonyl chloride- 2,2,3,3-d4

[0432] 6-(benzylsulfanyl)-2,3-dihydro-[l,4]dioxino[2,3-b]pyridine-2,2,3,3-d4 (0.66 g, 2.51 mmol) was dissolved in acetonitrile (10 mL), then added NCS (1.17 g, 8.79 mmol), acetic acid (1.0 mL), water (0.5 mL), stirred at room temperature for 1 hour. After monitoring the reaction by TLC, the reaction was quenched by adding a solution of sodium bisulfite (0.65 g) in water (20 mL), extracted with dichloromethane (25 mL x 3), the combined organic phase was washed with saturated brine (25 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated under reduced pressure to give the title compound (0.56 g, 93.23%) as a light yellow oil.

[0433] Fifth step: synthesis of tert-butyl 5-((2,3-dihydro-[l,4]dioxino[2,3-b]pyridin-6-yl- 2,2,3,3-d4) sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-2(lH)-carboxylate

[0434] To a reaction flask was added 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-2(lH)- carboxylic acid tert-butyl ester p-toluenesulfonic acid salt (0.9 g, 2.35 mmol), dichloromethane (10 mL) and triethylamine (0.59 g, 5.88 mmol) successively, 2,3-dihydro- [l,4]dioxino[2,3-b]pyridine-6-sulfonyl chloride-2,2,3,3-d4 (0.56 g, 2.35 mmol) was added with stirring, the reaction was stirred at room temperature for 45 minutes. After the reaction was completed by TLC monitoring, diluted with water (30 mL), extracted with dichloromethane (30 mL x 3), the organic layer was taken, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, the residue was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1 / 1), to give the title compound (0.27 g, 27.75%) as a yellow solid.

[0435] MS (ESI, pos.ion) m / z, 414.1 [M+H] + .

[0436] Step 6: Synthesis of 6-((3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)sulfonyl)- 2,3-dihydro-[l,4]dioxino[2,3-b]pyridine-2,2,3,3-d4 hydrochloride

[0437] To a reaction flask was added 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-2(lH)- carboxylic acid tert-butyl ester p-toluenesulfonic acid salt (0.9 g, 2.35 mmol), dichloromethane (10 mL) and triethylamine (0.59 g, 5.88 mmol) successively, 2,3-dihydro- [l,4]dioxino[2,3-b]pyridine-6-sulfonyl chloride-2,2,3,3-d4 (0.56 g, 2.35 mmol) was added with stirring, the reaction was stirred at room temperature for 45 minutes. After the reaction was completed by TLC monitoring, diluted with water (30 mL), extracted with dichloromethane (30 mL x 3), the organic layer was taken, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, the residue was separated and purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 1 / 1), to give the title compound (0.27 g, 27.75%) as a yellow solid.

[0438] MS (ESI, pos.ion) m / z, 414.1 [M+H] + .

[0439] Step 7: Synthesis of (R)-l-(5-((2,3-dihydro-[l,4]dioxino[2,3-b]pyridin-6-yl-2,2,3,3-d4) sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)-2-fluoro-3-hydroxy-2- phenylpropan-l-one

[0440] To a reaction flask was added (R)-2-fluoro-3-hydroxy-2-phenylpropanoic acid (0.12 g, 0.65 mmol), 6-((3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)sulfonyl)-2,3-dihydro- [l,4]dioxino[2,3-b]pyridine-2,2,3,3-d4 hydrochloride (0.24 g, 0.68 mmol), HOBt (0.13 g, 0.98 mmol), DMF (10 mL) and DIPEA (0.34 g, 2.60 mmol) successively, after stirring for 5 min, EDCI (0.19 g, 0.98 mmol) was added, the reaction was stirred at room temperature for 2.5 h. After the reaction was completed, water (60 mL) was added for dilution, extracted with ethyl acetate (30 mL x 2), the organic phase was collected, washed with water (30 mL x 2), saturated brine (30 mL) successively, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 3 / 7) to give the title compound (0.1 g, 32%) as a light yellow solid.

[0441] MS (ESI, pos.ion) m / z, 480.1 [M+H] + .

[0442] 1 H NMR (400 MHz, DMSO-d6) d 7.47 (s, 2H), 7.45 - 7.35 (m, 5H), 5.32 (t, J = 6.1 Hz, 1H), 4.45 (d, J = 14.8 Hz, 1H), 4.22 - 4.03 (m, 7H), 3.97 (d, J = 14.9 Hz, 1H), 3.69 (ddd, J = 19.2, 12.2, 6.1 Hz, 1H).

[0443] Example 119 2-Fluoro-3-hydroxy-2-(4-methoxyphenyl)-l-(5-(pyridine-2-sulfonyl)-3,4,5,6- tetrahydropyrrolo[3,4-c]pyrrol-2(lH)-yl)propan-l-one

[0444] First Step: Synthesis of ethyl 2-fluoro-2-(4-methoxyphenyl)acetate

[0445] To a reaction flask was added 4-methoxybenzeneboronic acid (8.21 g, 54.03 mmol), nickel(II) trifluoromethanesulfonate (0.68 g, 1.89 mmol), 4,4'-di-tert-butyl-2,2'-bipyridine (0.51 g, 1.89 mmol), triphenylphosphine (0.99 g, 3.78 mmol), potassium carbonate (15.69 g, 113.52 mmol), ethyl bromofluoroacetate (7 g, 37.84 mmol) and 1,4-dioxane (100 mL) successively, and the reaction was stirred at 100 °C for 15 hours. After the reaction was completed, it was filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10 / 1) to obtain the title compound (5.45 g, 67.87%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 7.41 (dd, J = 8.7, 1.6 Hz, 2H), 6.94 (d, J = 8.5 Hz, 2H), 5.73 (d, J = 47.9 Hz, 1H), 4.34 - 4.18 (m, 2H), 3.84 (s, 3H), 1.28 (t, J = 7.1 Hz, 3H); 19 F NMR (376 MHz, CDCl3) δ -174.67.

[0446] Second Step: Synthesis of Ethyl 2-fluoro-3-hydroxy-2-(4-methoxyphenyl)propanoate

[0447] To a solution of ethyl 2-fluoro-2-(4-methoxyphenyl)acetate (5.40 g, 25.45 mmol) in DMF (50 mL) was added paraformaldehyde (2.29 g, 76.35 mmol) and sodium ethoxide in ethanol (0.87 g, 2.54 mmol, 20% wt), and after the reaction was completed, water (150 mL) was added to the reaction solution, which was extracted with ethyl acetate (120 mL x 2), washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 4 / 1) to obtain the title compound (3.83 g, 62.14%) as a colorless oil.

[0448] 1 H NMR (400 MHz, CDCl3) δ 7.43 (d, J = 8.9 Hz, 2H), 6.92 (d, J = 8.7 Hz, 2H), 4.35 - 4.23 (m, 3H), 4.00 (dd, J = 15.6, 12.6 Hz, 1H), 3.81 (s, 3H), 1.30 (t, J = 7.1 Hz, 3H). 19 F NMR (376 MHz, CDCl3) δ -169.24.

[0449] Step 3: Synthesis of 2-fluoro-3-hydroxy-2-(4-methoxyphenyl)propanoic acid

[0450] To a solution of ethyl 2-fluoro-2-(4-methoxyphenyl)-3-hydroxypropanoate (3.5 g, 14.45 mmol) in methanol (24 mL) and water (6 mL) was added potassium carbonate (5.99 g, 43.35 mmol) and the reaction was stirred at room temperature. The reaction was stopped after TLC monitoring showed that the starting material had been completely consumed; the solvents were evaporated under reduced pressure, the residue was diluted with water (100 mL) and extracted with ethyl acetate (50 mL x 2), the organic phase was discarded; the aqueous phase was adjusted to pH = 1 with 2M hydrochloric acid and extracted with ethyl acetate (100 mL x 2), the combined organic phases were washed with saturated brine (80 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a white solid, which was the title compound (2.71 g, 87.57%). MS (ESI, neg. ion) m / z, 213.1 [M-H].

[0451] Step 4: Synthesis of 2-fluoro-3-hydroxy-2-(4-methoxyphenyl)-1-(5-(pyridine-2- sulfonyl)-3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)propan-1-one

[0452] To a reaction flask was added 2-fluoro-3-hydroxy-2-(4-methoxyphenyl)propanoic acid (0.2 g, 0.93 mmol), 2-(pyridin-2-ylsulfonyl)-1,2,3,4,5,6-hexahydropyrrolo[3,4- c]pyrrole dihydrochloride (0.33 g, 1.02 mmol), HOBt (0.19 g, 1.40 mmol), tetrahydrofuran (10 mL), DMF (10 mL) and DIPEA (0.48 g, 3.72 mmol) in sequence, and the reaction was stirred for 10 minutes before EDCI (0.23 g, 1.21 mmol) was added and the reaction was stirred at room temperature for 7 hours. The reaction was evaporated under reduced pressure, water (30 mL) was added to the residue, which was extracted with ethyl acetate (30 mL x 2), the organic phase was collected and washed with water (30 mL), saturated brine (30 mL) in sequence, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure; the residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate (v / v) = 1 / 4) to give the title compound (0.26 g, 62.23%) as a white solid. MS (ESI, pos. ion) m / z, 448.0 [M+H] + ;

[0453] 1H NMR (400 MHz, DMSO-d6) δ 8.71 (d, J = 4.7 Hz, 1H), 8.11 - 8.07 (m, 1H), 7.93 (d, J = 7.8 Hz, 1H), 7.71 - 7.64 (m, 1H), 7.30 (d, J = 8.4 Hz, 2H), 6.95 (d, J = 8.5 Hz, 2H), 5.26 (t, J = 6.1 Hz, 1H), 4.41 (d, J = 15.3 Hz, 1H), 4.27 - 4.02 (m, 7H), 3.96 (d, J = 15.3 Hz, 1H), 3.74 (s, 3H), 3.64 (ddd, J = 18.7, 12.3, 6.0 Hz, 1H).

[0454] Example 120 (S)-2-Fluoro-3-hydroxy-2-(4-methoxyphenyl)-1-(5-(pyridine-2-sulfonyl)- 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)propan-1-one and

[0455] Example 121 (R)-2-Fluoro-3-hydroxy-2-(4-methoxyphenyl)-1-(5-(pyridine-2-sulfonyl)- 3,4,5,6-tetrahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)propan-1-one

[0456] Example 119 was prepared by chromatography on a preparative chiral column: Chiralpak® IC column 5um*10mm*250mm (Waters supercritical fluid chromatography), sample dissolved in acetonitrile, 8 mL / min, column temperature 35°C, back pressure 100 bar; isocratic 70% methanol + 30% carbon dioxide, overlapping injections, fractions I (9.8-11.3 min) and II (11.4-13 min) were collected separately.

[0457] Example 120 Fraction I

[0458] MS (ESI, pos.ion) m / z, 448.1 [M+H] + ;

[0459] 1H NMR (400 MHz, DMSO-d6) δ 8.76 - 8.66 (m, 1H), 8.10 - 8.07 (m, 1H), 7.93 (d, J = 7.8 Hz, 1H), 7.67 (dd, J = 7.7, 4.7 Hz, 1H), 7.30 (d, J = 8.5 Hz, 2H), 6.95 (d, J = 8.4 Hz, 2H), 5.26 (t, J = 6.1 Hz, 1H), 4.40 (d, J = 15.4 Hz, 1H), 4.27 - 4.02 (m, 7H), 3.96 (d, J = 15.3 Hz, 1H), 3.74 (s, 3H), 3.64 (ddd, J = 18.6, 12.2, 5.6 Hz, 1H). 19 F NMR (376 MHz, DMSO-d6) δ -168.08.

[0460] Example 12 Component II

[0461] MS (ESI, pos.ion) m / z, 448.2 [M+H] + ;

[0462] 1 H NMR (400 MHz, DMSO-d6) δ 8.70 (dd, J = 4.8, 1.6 Hz, 1H), 8.11 - 8.07 (m, 1H), 7.93 (d, J = 7.8 Hz, 1H), 7.67 (dd, J = 7.7, 4.7 Hz, 1H), 7.30 (d, J = 8.5 Hz, 2H), 6.95 (d, J = 8.5 Hz, 2H), 5.25 (t, J = 6.1 Hz, 1H), 4.40 (d, J = 15.3 Hz, 1H), 4.27 - 4.03 (m, 7H), 3.96 (d, J = 15.5 Hz, 1H), 3.74 (s, 3H), 3.64 (ddd, J = 18.7, 12.2, 6.0 Hz, 1H). 19 F NMR (376 MHz, DMSO-d6) δ -168.08.

[0463] Biological test

[0464] Example A Pyruvate Kinase (PKR) Activity Test

[0465] 1. Reagents and Consumables

[0466] 2. Instruments

[0467] Centrifuge (Manufacturer: Eppendorf, Model: 5430)

[0468] Microplate Reader (Manufacturer: Perkin Elmer, Model: Envision)

[0469] 3. Experimental procedure

[0470] Prepare 1x Assay buffer: 50mM Tris-HCl pH=7.5, 5mM MgCl2, 100mM KCl, 0.03% BSA, 1mM DTT.

[0471] Dilute the compound in 100% DMSO to 100-fold final concentration working solution (5μL 10mM stock + 45μL 100% DMSO) in 96-well plate, working concentration is 1000μM.

[0472] Prepare DMSO solutions of the compound on 384-LDV plate: 10 concentrations of 1000μM, 333.33μM, 111.11μM, 37.04μM, 12.35μM, 4.12μM, 1.37μM, 0.457μM, 0.152μM and 0.0508μM (the highest concentration is 1000μM, and then diluted according to the 3-fold concentration gradient, 10 dose responses). Dilution method: transfer 15μL 1000μM compound DMSO solution from 96-well plate to 384-LDV echo plate Col1, add 10μL 100% DMSO to Col2 to Col9, transfer 5μL compound from Col1 to Col2, Col2 to Col3... Col8 to Col9 in turn by BROVO, add 10μL 100% DMSO to Col10 for backfill (high control and low control).

[0473] Transfer 50nL of the diluted compound to the 384-well detection plate (Corning; 4512) by ECHO. The assay plate also contains High control and Low control, both of which are filled with 50nL / well DMSO solution.

[0474] The final concentration of DMSO per well is 1%.

[0475] Prepare 2-fold final concentration (Final Conc. 0.5nM PKR2) enzyme solution with 1x Assay buffer, add 2.5μL per well, and replace the enzyme solution in the Low control well with 2.5μL of 1x Assay buffer; centrifuge at 1000rpm for 60 seconds, incubate at room temperature for 15 minutes.

[0476] • Prepare 2x final concentration (PKR: Final Conc. 78 mM PEP + 100 mM ADP / PKL / PKM2: 26 mM PEP + 100 mM ADP) phosphoenolpyruvate + ADP solution in lx Assay buffer, add 2.5 pL per well, centrifuge 1000 rpm for 60 seconds, incubate at room temperature for 15 min.

[0477] • Prepare CellTiter-Glo Luminescent Cell Viability Assay solution, add 10 pL per well, centrifuge 1000 rpm for 60 seconds, incubate at room temperature for the corresponding time.

[0478] • Envision reading.

[0479] • Formula:

[0480] Where: Reading is the signal value of the sample well; Low control: mean of Min control well signal values; High control: mean of Max control well signal values. According to the % increase in agonism fold of each gradient concentration well and its concentration, use the standard four-parameter fitting of ActivityBase XE Runner to draw the gradient curve of agonism fold, and calculate the AC 50 of the compound. The maximum fold % is the highest fold increase % calculated at any compound concentration, and the minimum fold % is the lowest fold increase % calculated at any compound concentration. The AC 50 of the compound is the concentration corresponding to the midpoint between the maximum and minimum values of the four-parameter logarithmic curve fitting, i.e. where the % increase in agonism fold along the four-parameter logarithmic curve fitting is the midpoint between the maximum fold % and the minimum fold % (fold % midpoint). The agonism fold value at 41 nM is the fold increase % at the compound concentration of 41 nM.

[0481] 4. The experimental results are shown in Table 1.

[0482] Table 1

[0483] “N / A” means blank or not tested.

[0484] Comparative compounds 1 and 2 are WO2018175474 Example 1 and Example 51, the specific structures are as follows:

[0485] The test results show that the compounds of the embodiments of the application have excellent agonistic activity on PKR in the in vitro recombinant PKR enzyme activity test. In terms of the agonistic fold on PKR at a concentration of 41 nM, Example 1 is superior to Comparative Compound 1, and Example 68 is superior to Comparative Compound 2, indicating that the compounds of the embodiments of the application have more excellent PKR agonistic activity.

[0486] Example B Red blood cell pyruvate kinase (PKR) activity experiment

[0487] 1. Reagents and consumables

[0488] 2. Instruments

[0489] Centrifuge (manufacturer: Eppendorf, model: 5430)

[0490] Microplate reader (manufacturer: MOLECULAR DEVICES, model: SpectraMax i3x)

[0491] 3. Experimental steps

[0492] • Compound stock solution: Take the compound out of the 4°C refrigerator, centrifuge it at 3000 rpm for 5 minutes using a centrifuge, and centrifuge most of the sample to the bottom of the EP tube. According to the given relative molecular mass, purity, and combined volume of the original EP tube used to store the drug, calculate the volume of DMSO to be used and ultrasonically agitate for 2 minutes to dissolve, so that the final concentration of each compound is 5 mM. The storage solution is stored in a 4°C refrigerator. (Materials: compound, 1.5 mL EP tube, record paper, pen, DMSO)

[0493] • Compound working solution: Take the storage solution and shake it well, then take samples in order to the 96-well plate and add DMSO for dilution, then blow and mix well to obtain 100 μl of working solution with a concentration of 1000 μM.

[0494] • Red blood cell acquisition: Take blood from the orbital venous plexus of male C57 mice, and place the blood sample in a 5 mL centrifuge tube anticoagulated with sodium heparin. Centrifuge at different speeds for different times, and discard the supernatant. Add 10 times the volume of isotonic PBS, centrifuge at the same speed for the same time, and discard the supernatant, repeat multiple times (see grouping conditions for details), until the supernatant is almost transparent, and the lower layer is red blood cells. (Materials: 5 mL heparin anticoagulation tube, 15 mL centrifuge tube, isotonic PBS, 1 mL / 5 mL pipette gun).

[0495] ● Compound dilution: Prepare the DMSO solution of the compound in 1.5 mL EP tubes: 10 concentrations of 1000 μM, 333.33 μM, 111.11 μM, 37.04 μM, 12.35 μM, 4.12 μM, 1.37 μM, 0.457 μM, 0.152 μM and 0.0508 μM (the highest concentration 1000 μM, and then diluted by 3 times concentration gradient, 10 dose responses). Dilution method: add 200 μL 100% DMSO to each EP tube 1-10, transfer 100 μL 1000 μM compound DMSO solution from the original tube to tube 1, add 100 μL compound DMSO solution in tube 1 to tube 2, add 100 μL compound DMSO solution in tube 2 to tube 3, add 100 μL compound DMSO solution in tube 3 to tube 4, add 100 μL compound DMSO solution in tube 4 to tube 5, add 100 μL compound DMSO solution in tube 5 to tube 6, add 100 μL compound DMSO solution in tube 6 to tube 7, add 100 μL compound DMSO solution in tube 7 to tube 8, add 100 μL compound DMSO solution in tube 8 to tube 9, add 100 μL compound DMSO solution in tube 9 to tube 10, and the dilution is completed.

[0496] ● Red blood cell incubation: Take 196 μL red blood cell suspension in a round-bottom 96-well plate, and then add 2 μL of the test compound (add 2 μL 100% DMSO to the control group) and 2 μL PEP in order of compound concentration, mix with a micro-vibration instrument, and incubate overnight at room temperature to allow the drug and cells to fully act. (Materials: round-bottom 96-well plate, 2.5 μL pipette, 100 / 300 μL pipette, sample addition slot, various test solutions, micro-vibration instrument). The final DMSO concentration in each well is 1%.

[0497] ● Red blood cell sample collection: centrifuge at 2500 rpm for 15 min to discard the supernatant, and freeze in a -80°C refrigerator.

[0498] ● Prepare the related substrate and probe solution of Pyruvate Kinase Assay Kit, add 50 μL mixed working solution and 50 μL sample diluent to each well, and incubate at room temperature for the corresponding time.

[0499] ● Enzyme reader reading.

[0500] ● Calculation formula: % Activity = (Reading - Low control) / (Low control) x 100

[0501] Wherein: Reading is the signal value of the sample well; Low control: signal value of the control well.

[0502] 4. The experimental results are shown in Table 2 below.

[0503] Table 2

[0504] The test results show that the compound of the present application has excellent PKR agonist activity in the PKR enzyme activity test of mouse-derived red blood cells.

[0505] Example C Whole Blood ATP Content Detection Experiment

[0506] I. Experimental Steps

[0507] 1. Dispensing Method

[0508] 1.1 Solvent group solvent: 5% DMSO + 5% Tween 80 + 20% PEG-400 + 70% physiological saline.

[0509] 1.2 Preparation of test substances (for example, 10 mL): accurately weigh an appropriate amount of raw drug (which needs to be converted according to the content of the raw drug) into a 50 mL centrifuge tube, add 0.5 mL of DMSO, ultrasonic until there are no obvious large particles, then add 0.5 mL of Tween 80 and blow evenly, then slowly add 2 mL of PEG-400 to make the drug evenly dispersed, blow evenly, ultrasonic, add 7 mL of physiological saline, ultrasonic, the solution is clear, shake or blow evenly before each time the drug is taken. The prepared drug solution is stored at 4°C.

[0510] 2. Clinical observation: observe the mice once a day, including appearance, fur, mental state, activity, trauma, etc.

[0511] 3. Diet and water: animals are allowed to eat and drink freely during the experiment.

[0512] 4. Sample collection: on the morning of D1, the Control group is given solvent, and the drug group mice are given the corresponding drug, which is given by gavage once a day, with a drug volume of 10 mL / kg, the drug amount is calculated according to the body weight, and the drug is given for 1 day. At each time point, 5 animals in the Control group and 5 animals in each group of the drug group are collected about 30 μL of EDTA anticoagulant whole blood, and the blood is collected from different sources. After blood collection, 10 μL of whole blood is immediately aliquoted into a 96-well plate, and the remaining whole blood is quickly frozen in a 1.5 mL EP tube, then transferred to a -80°C refrigerator for storage, and used for ATP detection. The blood collection time points are 0h, 24h, 48h, 72h, 168h.

[0513] II. Sample Preparation

[0514] 1. After the animal whole blood sample is taken out and placed on ice for 5 min, it is placed at room temperature for 5 min, and the sample is observed to see if it is completely thawed.

[0515] 2. Take out the CTG liquid and place it at room temperature for 30 min.

[0516] 3. After the incubation of step 3, the 96-well plate was allowed to stand for 5 min, and 100 uL of CTG solution was aspirated into each well by a multichannel pipette. The plate was mixed by a plate mixer at 400 rpm for 10 to 15 min at room temperature, and then the reading was taken by a microplate reader.

[0517] 4. The ATP standard curve samples were treated as described above.

[0518] III. Sample treatment / incubation

[0519] 1. After the 10 uL whole blood sample was completely thawed, 190 uL of lysis solution was added to make the red blood cells fully lysed and diluted. The sample was mixed by a plate mixer at 300 rpm for 1 to 2 min at room temperature, and then the mixing and lysis were completed.

[0520] 2. The 96-well plate was allowed to stand for 5 min at room temperature, and 150 uL of CTG solution was aspirated into each well by a multichannel pipette. After mixing by shaking, 50 uL of the whole blood diluted sample was aspirated into each well by a multichannel pipette, and then mixed by shaking. The plate was mixed by a plate mixer at 300 rpm for 10 to 15 min at room temperature, and then the reading was taken by a microplate reader.

[0521] 3. The ATP standard curve samples were treated as described above.

[0522] IV. Microplate reader reading and ATP increase calculation

[0523] 1. The corresponding template program was selected for reading.

[0524] 2. The ATP content of each well was calculated according to the corresponding standard curve.

[0525] 3. The calculation formula was: % ATP increase = (Reading - Baseline reading) / (Baseline reading) x 100

[0526] Wherein: Reading is the signal value of the sample well corresponding to the ATP content; Baseline reading1 is the signal value of the initial (baseline) sample well corresponding to the ATP content.

[0527] The results are shown in Tables 3 and 4

[0528] Table 3

[0529] Table 4

[0530] The comparative compounds 1 and 2 are Example 1 and Example 51 of the patent WO2018175474, and the specific structures are as follows:

[0531] Conclusion: The test results show that the embodiment of the present application can significantly increase the whole blood ATP content in mice after single oral administration, indicating that the embodiment of the present application can increase the synthesis of ATP by activating PKR in vivo. The increase of whole blood ATP content of Example 1 at a dose of 100 mg / kg is about twice that of Comparative Compound 1 at a dose of 200 mg / kg, and the duration of drug efficacy is longer, the increase of whole blood ATP content of Example 68 at a dose of 10 mg / kg is about twice that of Comparative Compound 2 at the same dose, and the duration of drug efficacy is longer, indicating that the compound of the embodiment of the present application has more optimal and more persistent in vivo efficacy.

[0532] In the description of the present specification, the description of the terms "one embodiment", "some embodiments", "some embodiments", "example", "specific example" or "some examples" and the like means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, different embodiments, embodiments or features of different embodiments, embodiments or examples described in the present specification can be combined and combined by those skilled in the art without contradiction.

[0533] Although the embodiments of the present application have been shown and described above, it is understood that the above embodiments are exemplary and cannot be construed as limiting the present application, and those skilled in the art can make changes, modifications, replacements and variations to the above embodiments without departing from the principles and spirit of the present application within the scope of the present application, the scope of the present application is defined by the claims and their equivalents.

Claims

A compound that is a compound of Formula (I), or a stereoisomer, tautomer, nitroso, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of a compound of Formula (I), wherein R 1 is C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 1-6 alkoxy, C 1-6 alkyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, C 6-10 aryl or 5-12 membered heteroaryl, wherein the C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 1-6 alkoxy, C 1-6 alkyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, C 6-10 aryl and 5-12 membered heteroaryl are each independently optionally substituted with 1, 2, 3, 4, or 5 substituents selected from the group consisting of D, -OH, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 deuteroalkoxy, C 1-6 haloalkoxy, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl, 5-12 membered heteroaryl, -OR a , -C(=O)R a , -C(=O)OR a , -NR b R c , -C(=O)NR b R c , -NR b C(=O)R c , -S(O)R a , -S(O)2R a , -S(O)2NR b R c , and -NR b S(O)2R c ; R 2 is H, D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 1-6 alkoxyC 1-6 alkyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, C 6-10 aryl or 5-12 membered heteroaryl, wherein said C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 1-6 alkoxyC 1-6 alkyl, C 3-12 cycloalkyl, 3-12 membered heterocyclyl, C 6-10 aryl and 5-12 membered heteroaryl are each independently optionally substituted with 1, 2, 3, 4, or 5 groups selected from D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxyC 1-6 alkoxy, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, -OR d , -C(=O)R d , -C(=O)OR d , -NR e R f , -C(=O)NR e R f , -NR e C(=O)R f , -S(O)R d , -S(O)2R d , -S(O)2NR e R f , and -NR e S(O)2R f substituted by one or more substituents selected from the group consisting of R 3 and R 4 each independently H, D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 1-6 alkylamino, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl, and 3-8 membered heterocyclyl, wherein said C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 1-6 alkylamino, C 1-6 aminoalkyl, C 1-6 cyanoalkyl, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl, and 3-8 membered heterocyclyl are each independently optionally substituted with 1, 2, 3, or 4 substituents selected from D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, oxo, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, and 3-6 membered heterocyclyl; Or, R 3 R 4 Together with the carbon atoms attached to them, they form C 3-8 Cycloalkyl or heterocyclic group consisting of 3-8 atoms, wherein the C 3-8 The cycloalkyl group and the heterocyclic group consisting of 3-8 atoms are each independently and optionally surrounded by 1, 2 or 3 atoms selected from D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, oxo, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy and C 1-6 Substituents of haloalkoxy groups; R a , R b , R c , R d , R e , and R f are each independently H, D, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl, or 5-12 membered heteroaryl, wherein the C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl, and 5-12 membered heteroaryl are each independently optionally substituted with 1, 2, 3, or 4 substituents selected from D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, and C 1-6 haloalkoxy. The compound according to claim 1, wherein R 1 is C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 haloalkyl, C 1-4 aminoalkyl, C 1-4 cyanoalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxyC 1-4 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl or 5-10 membered heteroaryl, wherein said C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 haloalkyl, C 1-4 aminoalkyl, C 1-4 cyanoalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxyC 1-4 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl and 5-10 membered heteroaryl are each independently optionally substituted with 1, 2, 3, 4, or 5 substituents selected from the group consisting of D, -OH, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 deuteroalkoxy, C 1-4 haloalkoxy, C 3-6 cycloalkyl, 3-6 membered heterocyclyl, C 6-10 aryl, 5-10 membered heteroaryl, -OR a , -C(=O)R a , -C(=O)OR a , -NR b R c , -C(=O)NR b R c , -NR b C(=O)R c , -S(O)R a , -S(O)2R a , -S(O)2NR b R c , and -NR b S(O)2R c . The compound according to claim 1 or 2, wherein, R 1 methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, vinyl, ethynyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylen, ethoxymethylen, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, The terms methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, vinyl, ethynyl, -CH2F, -CHF2, -CH2CF2, -CH2CF3, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl, aziroxybutyl, pyrrolyl, tetrahydrofuranyl, piperidinyl, piperazine, morpholinyl, phenyl, naphthyl, pyrrolyl, pyrazolyl, imidazoleyl, triazolyl, tetraazolyl, furanyl, thiophenyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyrazinyl each independently optionally substituted with 1, 2, 3, 4, or 5 substituents selected from D, -OH, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, i-propoxy, deuterated methoxy, deuterated ethoxy, difluoromethoxy, trifluoromethoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, oxopyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, indenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, -OR a , -C(=O)R a , -C(=O)OR a , -NR b R c , -C(=O)NR b R c , -NR b C(=O)R c , -S(O)R a , -S(O)2R a , -S(O)2NR b R c , and -NR b S(O)2R c . The compound according to any one of claims 1-3, wherein R 2 is H, D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, oxo, -SF5, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 aminoalkyl, C 1-4 cyanoalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxyC 1-4 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl or 5-10 membered heteroaryl, wherein said C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 aminoalkyl, C 1-4 cyanoalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxyC 1-4 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl and 5-10 membered heteroaryl are each independently optionally substituted with 1, 2, 3, 4, or 5 groups selected from D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, oxo, -SF5, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, hydroxyC 1-4 alkoxy, C 3-6 cycloalkyl, 3-6 membered heterocyclyl, -OR d , -C(=O)R d , -C(=O)OR d , -NR e R f , -C(=O)NR e R f , -NR e C(=O)R f , -S(O)R d , -S(O)2R d , -S(O)2NR e R f , and -NR e S(O)2R f substituted by one or more substituents selected from the group consisting of The compound according to any one of claims 1-4, wherein R 2 H, D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, ethenyl, ethynyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, i-propoxy, difluoromethoxy, trifluoromethoxy, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetanyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, indenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, The terms methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, vinyl, ethynyl, -CH2F, -CHF2, -CH2CF2, -CH2CF3, methoxy, ethoxy, isopropoxy, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxacyclobutyl, azacyclobutyl, pyrrolyl, tetrahydrofuranyl, piperidinyl, piperazine, morpholinyl, phenyl, indole, naphthyl, pyrrolyl, pyrazolyl, imidazoleyl, triazolyl, tetraazolyl, furanyl, thiophenyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyrazinyl each independently optionally substituted with 1, 2, 3, 4, or 5 substituents selected from the group consisting of D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, oxo, -SF5, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, i-propoxy, difluoromethoxy, trifluoromethoxy, hydroxymethoxy, hydroxyethoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, -OR d , -C(=O)R d , -C(=O)OR d , -NR e R f , -C(=O)NR e R f , -NR e C(=O)R f , -S(O)R d , -S(O)2R d , -S(O)2NR e R f , and -NR e S(O)2R f . The compound according to any one of claims 1-5, wherein R 3 and R 4 each independently H, D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 haloalkyl, C 1-4 haloalkoxy, C 1-4 alkylamino, C 1-4 aminoalkyl, C 1-4 cyanoalkyl, C 1-4 hydroxyalkyl, C 3-6 cycloalkyl, and 3-6 membered heterocyclyl, wherein said C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, C 1-4 haloalkyl, C 1-4 haloalkoxy, C 1-4 alkylamino, C 1-4 aminoalkyl, C 1-4 cyanoalkyl, C 1-4 hydroxyalkyl, C 3-6 cycloalkyl, and 3-6 membered heterocyclyl are each independently optionally substituted with 1, 2, 3, or 4 substituents selected from D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, oxo, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 haloalkyl, C 1-4 haloalkoxy, C 3-6 cycloalkyl, and 3-6 membered heterocyclyl; Or, R 3 R 4 Together with the carbon atoms attached to them, they form C 3-6 Cycloalkyl or heterocyclic group consisting of 3-6 atoms, wherein the C 3-6 The cycloalkyl group and the heterocyclic group consisting of 3-6 atoms are each independently and optionally surrounded by 1, 2 or 3 atoms selected from D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, oxo, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy and C 1-4 The substituents of the haloalkoxy group are replaced. The compound according to any one of claims 1-6, wherein R 3 and R 4 each independently H, D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, ethenyl, ethynyl, methoxy, ethoxy, i-propoxy, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, difluoromethoxy, trifluoromethoxy, N-methylamino, N-ethylamino, N,N-dimethylamino, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, or morpholinyl, wherein each of said methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, ethenyl, ethynyl, methoxy, ethoxy, i-propoxy, -CH2F, -CHF2, -CH2CF2, -CH2CF3, N-methylamino, N-ethylamino, N,N-dimethylamino, aminomethyl, aminoethyl, cyanomethyl, cyanoethyl, hydroxymethyl, hydroxyethyl, methoxymethylene, ethoxymethylene, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, and morpholinyl is independently optionally substituted with 1, 2, 3, or 4 substituents selected from D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, oxo, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, methoxy, ethoxy, i-propoxy, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, difluoromethoxy, trifluoromethoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, and morpholinyl; or R 3 or R 4 and the carbon atom to which they are attached together form a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, or morpholinyl, wherein each of said cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, and morpholinyl is independently optionally substituted with 1, 2, or 3 substituents selected from the group consisting of D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, oxo, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, i-propoxy, difluoromethoxy, and trifluoromethoxy. The compound according to any one of claims 1-7, wherein R a , R b , R c , R d , R e , and R f are each independently H, D, C 1-4 alkyl, C 1-4 haloalkyl, C 3-6 cycloalkyl, 3-6 membered heterocyclyl, C 6-10 aryl, or 5-10 membered heteroaryl, wherein the C 1-4 alkyl, C 1-4 haloalkyl, C 3-6 cycloalkyl, 3-6 membered heterocyclyl, C 6-10 aryl, and 5-10 membered heteroaryl are each independently optionally substituted with 1, 2, 3, or 4 substituents selected from D, -F, -CI, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, and C 1-4 haloalkoxy. The compound according to any one of claims 1-8, wherein R a , R b , R c , R d , R e , and R f are each independently H, D, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, indenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, or pyridazinyl, wherein each of said methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CH2CF2, -CH2CF3, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, indenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, and pyridazinyl is independently optionally substituted with 1, 2, 3, or 4 substituents selected from the group consisting of D, -F, -Cl, -Br, -I, -OH, -NH2, -CN, -SH, -SF5, oxo, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, -CH2F, -CHF2, -CF3, -CH2CF2, -CH2CF3, methoxy, ethoxy, i-propoxy, difluoromethoxy, and trifluoromethoxy. The compound according to any one of claims 1 to 9, which is a compound of formula (I-a) or (I-b), or a stereoisomer, a tautomer, a nitroso, a hydrate, a solvate, a metabolite, a pharmaceutically acceptable salt or a prodrug of a compound of formula (I-a) or (I-b), wherein, R 1 , R 2 , R 3 and R 4 each have the meaning as defined in any one of claims 1 to 9. A compound according to any one of claims 1-10, having the structure: ###0001### or a stereoisomer, tautomer, nitroso, hydrate, solvate, metabolite, pharmaceutically acceptable salt, or prodrug thereof, ​ A pharmaceutical composition comprising a compound according to any one of claims 1-11; optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable adjuvant. Use of a compound according to any one of claims 1-11 or a pharmaceutical composition according to claim 12 in the manufacture of a medicament for preventing, treating or alleviating a PKR-related disease. Use according to claim 13, wherein, The PKR-related disease is thalassemia, pyruvate deficiency anemia, anemia associated with myelodysplastic syndrome (MDS), sickle anemia, megaloblastic anemia, aplastic anemia, iron deficiency anemia, hemolytic anemia, sickle cell disease, hereditary non-spherocytic hemolytic anemia, hereditary spherocytosis, hereditary elliptocytosis, abetalipoproteinemia, paroxysmal nocturnal hemoglobinuria, acquired hemolysis, tumor-related anemia or chronic disease-related anemia.

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