A PI3Kδ inhibitor and its uses

Abstract

The present invention relates to a novel PI3Kδ inhibitor, comprising a compound of formula (I) or a pharmaceutically acceptable salt, solvate, polymorph, ester, acid, isomer, metabolite or prodrug thereof. The present invention also provides the use of a PI3Kδ inhibitor in the preparation of a medicament for inhibiting PI3Kδ kinase activity or treating a disease or disorder associated with PI3Kδ kinase activity.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to compounds as selective PI3Kδ inhibitors, and methods and uses for using such compounds to inhibit PI3Kδ kinase activity and treat diseases or disorders related to the inhibition of PI3Kδ kinase activity. Background Art

[0002] PI3K is called phosphatidylinositol-3-kinase, and can be divided into three categories according to structural and substrate differences: class I, class II, and class III. Class I PI3K utilizes phosphatidylinositol 4,5-bisphosphate to generate phosphatidylinositol 3,4,5-trisphosphate (PIP3), and it consists of a catalytic subunit and a regulatory subunit, and can be further divided into two families, namely IA and IB. Among them, IB PI3K is formed by coupling the catalytic subunit P110γ with the p101 or p84 regulatory subunit. IA PI3K can be further divided into three subtypes: PI3Kα, PI3Kβ, and PI3Kδ, and their catalytic subunits are p110α, p110β, and p110δ respectively, and they are coupled with one of the five regulatory subunits p85α, p55α, p50α, p85β, and p55γ to form IA PI3K. PI3Kα and PI3Kβ are widely expressed in various cells, while PI3Kδ is mainly expressed in hematopoietic cells and immune cells, and is a key mediator of B cell receptor signaling, and is closely related to the survival, migration, and activation of B cells. As a key signaling molecule in the occurrence and development of B cell-related autoimmune diseases and malignant hematological tumors, it has thus become a potentially effective target for treating these diseases.

[0003] Idelalisib, Copanlisib, and Duvelisib are three approved PI3Kδ inhibitors. Among them, the PI3Kδ inhibitor Idelalisib developed by Gilead has extremely poor tolerance, has 4 black box warnings, and side effects are almost all over the body, such as the liver (elevated transaminases), digestive tract (diarrhea, colitis, intestinal perforation), immune system (pneumonia, infection), skin (rash), and blood (neutropenia), and in clinical practice, there is even a treatment discontinuation rate of more than 50%, and it finally withdrew from the market. Duvelisib has 1 black box warning.

[0004] Although the emergence of PI3Kδ inhibitors has opened up a new situation in the treatment of malignant hematological tumors, due to the relatively serious side effects of existing PI3Kδ inhibitors, there is an urgent need to develop new PI3Kδ inhibitors with high potency, high selectivity, and low toxicity. Summary of the Invention

[0005] One object of the present invention is to provide a class of PI3Kδ inhibitors, including compounds of formula (I) or pharmaceutically acceptable salts, solvates, polymorphs, esters, acids, isomers, metabolites or prodrugs thereof:

[0006]

[0007] Wherein,

[0008] Each R1 is independently selected from halogen and C1-C4 alkyl; m is 1 or 2;

[0009] R2 is selected from C3-C6 cycloalkyl, C3-C6 cycloalkyl C1-C4 alkyl, and phenyl or pyrazolyl optionally substituted by halogen or methyl;

[0010] R3 and R4 are each independently selected from H and C1-C4 alkyl;

[0011] R5 is selected from C3-C8 branched alkyl, C1-C8 haloalkyl, C1-C4 alkoxy C1-C4 alkyl, C2-C6 hydroxyalkyl, C3-C6 cycloalkyl C1-C4 alkyl, 4-6 membered heterocycloalkyl, and 4-6 membered heterocycloalkyl C1-C4 alkyl.

[0012] In a preferred embodiment, R5 is selected from isopropyl, isobutyl, pentan-3-yl, monohaloalkyl (preferably haloethyl), dihaloalkyl (preferably dihalomethyl), 1-ethoxy-ethyl, hydroxyethyl, cyclopropylmethyl, oxetanyl, tetrahydropyranyl and morpholinylethyl. Particularly preferably, R5 is difluoromethyl.

[0013] In another preferred embodiment, R1 is selected from fluorine and methyl.

[0014] In yet another preferred embodiment, R2 is selected from cyclopropyl, cyclopropylmethyl, fluorophenyl, and N-methyl-pyrazolyl.

[0015] In other preferred embodiments, one of R3 and R4 is H and the other is methyl or ethyl.

[0016] On the other hand, the present application also provides a pharmaceutical composition, which comprises a compound of formula (I) or a pharmaceutically acceptable salt, solvate, polymorph, ester, acid, isomer, metabolite or prodrug thereof, and a pharmaceutically acceptable diluent or carrier, and optionally other active ingredients.

[0017] In other aspects, the present application also relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, polymorph, ester, acid, isomer, metabolite or prodrug thereof in the preparation of a drug for inhibiting PI3Kδ kinase activity or treating a disease or disorder associated with PI3Kδ kinase activity.

[0018] The disease or disorder is selected from chronic lymphocytic leukemia, non-Hodgkin lymphoma, Hodgkin lymphoma, mantle cell lymphoma, chronic obstructive pulmonary disease, rheumatoid arthritis, systemic lupus erythematosus, and asthma.

[0019] The present invention also relates to a method for inhibiting PI3Kδ kinase activity or treating a disease or disorder related to PI3Kδ kinase activity, which comprises administering to a patient a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, polymorph, ester, acid, isomer, metabolite, or prodrug thereof, or a pharmaceutical composition comprising the same. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 Shows the effects of Compound 12, Compound 24, Linperlisib of the present invention, and vehicle administration on the body weight of mice in a DOHH2 mouse xenograft tumor model.

[0021] Figure 2 Shows the effects of Compound 12, Compound 24, Linperlisib of the present invention, and vehicle administration on the tumor size of a DOHH2 mouse xenograft tumor model.

[0022] Figure 3 Shows the effects of Compound 24 of the present invention, Comparative Compound 1, and vehicle administration on the body weight of mice in an MC38 mouse xenograft tumor model.

[0023] Figure 4 Shows the effects of Compound 24 of the present invention, Comparative Compound 1, and vehicle administration on the tumor size of an MC38 mouse xenograft tumor model.

[0024] Figure 5 Shows the effects of Compound 24 of the present invention, Comparative Compound 1, and vehicle administration on the tumor weight of an MC38 mouse xenograft tumor model. DETAILED DESCRIPTION OF THE INVENTION

[0025] Term

[0026] Unless otherwise defined, all scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art to which the claimed subject matter pertains.

[0027] Unless otherwise specified, the present invention employs conventional methods such as mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA technology, and pharmacology within the scope of those skilled in the art. Unless specific definitions are provided, chemical nomenclature, laboratory operations, and techniques related to analytical chemistry, synthetic organic chemistry, and medical and pharmaceutical chemistry described herein are known to those skilled in the art. Generally, the aforementioned techniques and procedures can be implemented by conventional methods well-known in the art and described in various general and more specific documents, which are cited and discussed in this specification.

[0028] The term "alkyl" refers to an aliphatic hydrocarbon group, which can be a branched or straight-chain alkyl. Depending on the structure, an alkyl can be a monovalent group or a divalent group (i.e., an alkylene). In the present invention, the alkyl is preferably an alkyl having 1-8 carbon atoms, more preferably a "lower alkyl" having 1-6 carbon atoms, and even more preferably an alkyl having 1-4 carbon atoms. Typical alkyls include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, etc. It should be understood that the "alkyl" mentioned herein includes all possible configurations and conformations of the alkyl, for example, the "propyl" mentioned herein includes n-propyl and isopropyl, the "butyl" includes n-butyl, isobutyl, and tert-butyl, and the "pentyl" includes n-pentyl, isopentyl, neopentyl, tert-pentyl, and pent-3-yl, etc.

[0029] The term "alkoxy" refers to -O-alkyl, where the alkyl is as defined herein. Typical alkoxys include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, etc.

[0030] The term "cycloalkyl" refers to a monocyclic or polycyclic group that contains only carbon and hydrogen. Cycloalkyls include groups having 3-12 ring atoms. Depending on the structure, a cycloalkyl can be a monovalent group or a divalent group (e.g., a cycloalkylene). In the present invention, the cycloalkyl is preferably a cycloalkyl having 3-8 carbon atoms, more preferably a "lower cycloalkyl" having 3-6 carbon atoms. Examples of cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, and adamantyl.

[0031] The term "aryl" refers to a planar ring having a delocalized π-electron system and containing 4n + 2 π electrons, where n is an integer. The aryl ring can be composed of five, six, seven, eight, nine, or more than nine atoms. The aryl can be optionally substituted. The term "aryl" includes carbocyclic aryl (e.g., phenyl) and heteroaryl (or "heteroaromatic" or "heteroaromatic") groups (e.g., pyridine). The term includes monocyclic or fused polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) groups.

[0032] As used herein, the term "aryl" refers to an aromatic ring in which each ring-forming atom is a carbon atom. The aryl ring can be composed of five, six, seven, eight, nine or more than nine atoms. The aryl can be optionally substituted. Examples of aryl include, but are not limited to, phenyl, naphthyl, phenanthryl, anthryl, fluorenyl, and indenyl. Depending on the structure, the aryl can be a monovalent group or a divalent group (i.e., arylene).

[0033] The term "aryloxy" refers to -O-aryl, where aryl is as defined herein.

[0034] The term "heteroaryl" refers to an aryl that includes one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur. The N-containing "heteroaryl" moiety refers to an aromatic ring in which at least one backbone atom is a nitrogen atom. Depending on the structure, the heteroaryl can be a monovalent group or a divalent group (i.e., heteroarylene). Examples of heteroaryl include, but are not limited to, pyridyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, indazolyl, indazinyl, phthalazinyl, pyridazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl, naphthyridinyl, and furanopyridyl, etc.

[0035] As used herein, the term "heteroalkyl" refers to an alkyl as defined herein in which one or more backbone chain atoms are heteroatoms, such as oxygen, nitrogen, sulfur, silicon, phosphorus, or a combination thereof. The heteroatom(s) can be located at any position within the heteroalkyl or at the position where the heteroalkyl is attached to the rest of the molecule.

[0036] As used herein, the term "heterocycloalkyl" or "heterocyclic group" means a non-aromatic ring in which one or more of the atoms constituting the ring are heteroatoms selected from nitrogen, oxygen, and sulfur. The heterocycloalkyl ring can be composed of three, four, five, six, seven, eight, nine, or more than nine atoms. The heterocycloalkyl ring can be optionally substituted. Examples of heterocycloalkyl include, but are not limited to, lactam, lactone, cyclic imine, cyclic thioimine, cyclic carbamate, tetrahydrothiopyran, 4H-pyran, tetrahydropyran, piperidine, 1,3-dioxin, 1,3-dioxane, 1,4-dioxin, 1,4-dioxane, piperazine, 1,3-oxathiane, 1,4-oxathiadiene, 1,4-oxathiane, tetrahydro-1,4-thiazine, 2H-1,2-oxazine, maleimide, succinimide, barbituric acid, thiobarbituric acid, dioxopiperazine, hydantoin, dihydrouracil, morpholine, trioxane, hexahydro-1,3,5-triazine, tetrahydrothiophene, tetrahydrofuran, pyrroline, pyrrolidine, imidazolidine, pyrrolidone, pyrazoline, pyrazolidine, imidazoline, imidazolidine, 1,3-dioxolene, 1,3-dioxolane, 1,3-dithiolene, 1,3-dithiolane, isoxazoline, isoxazolidine, oxazoline, oxazolidine, oxazolidinone, thiazoline, thiazolidine, and 1,3-oxathiolane. Depending on the structure, heterocycloalkyl can be a monovalent group or a divalent group (i.e., heterocycloalkylidene).

[0037] The term "halo" or "halogen" means fluorine, chlorine, bromine, and iodine.

[0038] The terms "haloalkyl", "haloalkoxy", and "haloheteroalkyl" include structures of alkyl, alkoxy, or heteroalkyl in which at least one hydrogen is replaced by a halogen atom. In certain embodiments, if two or more hydrogen atoms are replaced by halogen atoms, the halogen atoms are the same or different from each other.

[0039] The term "amino" means the -NH2 group.

[0040] The term "hydroxy" means the -OH group.

[0041] The term "cyano" means the -CN group.

[0042] The term "ester group" means a chemical moiety having the formula -COOR, where R is selected from alkyl, cycloalkyl, aryl, heteroaryl (attached through a ring carbon), and heterocyclic group (attached through a ring carbon).

[0043] The term "amide group" or "acylamino" means -NR-CO-R', where R and R' are each independently hydrogen or alkyl.

[0044] The term "aminoacyl" or "amineacyl" means the -CO-NH2 group.

[0045] The term "alkylaminoacyl" or "alkylamido" refers to a -CO-NH-R group, where R is an alkyl group as defined herein.

[0046] The term "optionally" means that one or more of the events described hereinafter may or may not occur, and includes both the events that occur and the events that do not occur. The term "optionally substituted" or "substituted" means that the group mentioned may be substituted by one or more additional groups, and the additional groups are each independently selected from alkyl, cycloalkyl, aryl, heteroaryl, heterocyclic, hydroxy, alkoxy, cyano, halogen, amido, nitro, haloalkyl, amino, mesyl, alkylcarbonyl, alkoxycarbonyl, heteroarylalkyl, heterocycloalkylalkyl, aminoacyl, amino protecting group, etc. Among them, the amino protecting group is preferably selected from pivaloyl, tert-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyl, p-methoxybenzyl, allyloxycarbonyl, trifluoroacetyl, etc.

[0047] The term "tyrosine protein kinase (TPK)" used herein is a class of kinases that catalyze the transfer of the γ-phosphate on ATP to the tyrosine residue of a protein, and can catalyze the phosphorylation of tyrosine residues of a variety of substrate proteins, and plays an important role in cell growth, proliferation and differentiation.

[0048] The terms "inhibition", "inhibited" or "inhibitor" of a kinase used herein mean that the phosphotransferase activity is inhibited.

[0049] The "metabolite" of a compound disclosed herein is a derivative of a compound formed when the compound is metabolized. The term "active metabolite" refers to a bioactive derivative of a compound formed when the compound is metabolized. As used herein, the term "being metabolized" refers to the sum total of processes by which a particular substance is altered by an organism (including but not limited to hydrolysis reactions and enzyme-catalyzed reactions such as oxidation reactions). Thus, enzymes can effect specific structural transformations to compounds. For example, cytochrome P450 catalyzes various oxidation and reduction reactions, while uridine diphosphate glucuronosyltransferase catalyzes the conversion of an activated glucuronic acid molecule to an aromatic alcohol, an aliphatic alcohol, a carboxylic acid, an amine, and a free sulfhydryl group. Further information on metabolism can be obtained from 《The Pharmacological Basis of Therapeutics》, Ninth Edition, McGraw-Hill (1996). The metabolites of a compound disclosed herein can be identified by administering the compound to a host and analyzing tissue samples from the host, or by incubating the compound with hepatocytes in vitro and analyzing the resulting compounds. Both methods are known in the art. In some embodiments, the metabolite of a compound is formed through an oxidation process and corresponds to the corresponding hydroxy-containing compound. In some embodiments, the compound is metabolized to a pharmaceutically active metabolite. As used herein, the term "regulate" means to interact directly or indirectly with a target to change the activity of the target, and by way of example only, includes enhancing the activity of the target, inhibiting the activity of the target, restricting the activity of the target, or prolonging the activity of the target.

[0050] The IC used herein 50 refers to the amount, concentration, or dose of a particular test compound that produces 50% inhibition of the maximal effect in an assay measuring such an effect.

[0051] The EC used herein 50 refers to the dose, concentration, or amount of a compound that produces a dose-dependent response that is 50% of the maximal expression of a particular response induced, stimulated, or enhanced by the particular test compound.

[0052] The GI used herein 50 refers to the drug concentration required to inhibit 50% of cell growth, i.e., the drug concentration at which the growth of 50% of cells (such as cancer cells) is inhibited or controlled.

[0053] Novel kinase inhibitor of the present invention

[0054] The present invention provides a PI3Kδ inhibitor, comprising a compound of formula (I) or a pharmaceutically acceptable salt, solvate, polymorph, ester, acid, isomer, metabolite, or prodrug thereof:

[0055]

[0056] Among them,

[0057] each R1 is independently selected from halogen and C1-C4 alkyl; m is 1 or 2;

[0058] R2 is selected from C3-C6 cycloalkyl, C3-C6 cycloalkyl C1-C4 alkyl, and phenyl or pyrazolyl optionally substituted by halogen or methyl;

[0059] each of R3 and R4 is independently selected from H and C1-C4 alkyl;

[0060] R5 is selected from C3-C8 branched alkyl, C1-C8 haloalkyl, C1-C4 alkoxy C1-C4 alkyl, C2-C6 hydroxyalkyl, C3-C6 cycloalkyl C1-C4 alkyl, 4-6 membered heterocycloalkyl, and 4-6 membered heterocycloalkyl C1-C4 alkyl.

[0061] In a preferred embodiment, R5 is selected from isopropyl, isobutyl, pent-3-yl, monohaloalkyl (preferably haloethyl), dihaloalkyl (preferably dihalomethyl), 1-ethoxy-ethyl, hydroxyethyl, cyclopropylmethyl, oxetanyl, tetrahydropyranyl and morpholinylethyl. Particularly preferably, R5 is difluoromethyl.

[0062] In another preferred embodiment, R1 is selected from fluorine and methyl.

[0063] In yet another preferred embodiment, R2 is selected from cyclopropyl, cyclopropylmethyl, fluorophenyl and N-methyl-pyrazolyl.

[0064] In other preferred embodiments, one of R3 and R4 is H and the other is methyl or ethyl.

[0065] For each variable, any combination of the above groups is also contemplated herein. It is understood that: the substituents and substitution patterns on the compounds provided herein can be selected by those skilled in the art so as to provide compounds that are chemically stable and can be synthesized using techniques known in the art as well as the techniques described herein.

[0066] The pharmaceutically acceptable salts, solvates, polymorphs, esters, acids, isomers, metabolites or prodrugs of this compound are also described herein.

[0067] In a preferred embodiment, the present invention relates to the compounds in Table 1 below or their pharmaceutically acceptable salts, solvates, polymorphs, esters, acids, isomers, metabolites or prodrugs.

[0068] Table 1

[0069]

[0070]

[0071]

[0072] In additional or further embodiments, the compounds described herein are metabolized in vivo in an organism in need thereof to produce metabolites, and the metabolites so produced are then used to produce a desired effect, including a desired therapeutic effect.

[0073] The compounds described herein can be made and / or used as pharmaceutically acceptable salts. Types of pharmaceutically acceptable salts include, but are not limited to: (1) acid addition salts, formed by reacting the free base form of the compound with a pharmaceutically acceptable inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, metaphosphoric acid, etc.; or with an organic acid, such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, malic acid, citric acid, succinic acid, maleic acid, tartaric acid, fumaric acid, trifluoroacetic acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, 4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, 2-naphthalenesulfonic acid, tert-butylacetic acid, glucoheptonic acid, 4,4'-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, dodecylsulfuric acid, gluconic acid, glutamic acid, salicylic acid, hydroxy-naphthoic acid, stearic acid, mucic acid, etc.; (2) base addition salts, formed when an acidic proton in the parent compound is replaced by a metal ion, such as an alkali metal ion (e.g., lithium, sodium, potassium), an alkaline earth metal ion (e.g., magnesium or calcium), or an aluminum ion; or coordinated with an organic or inorganic base, acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, trimethylamine, N-methylglucosamine, etc.; acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, etc.

[0074] The corresponding counterions of the pharmaceutically acceptable salts can be analyzed and identified using various methods, including but not limited to ion exchange chromatography, ion chromatography, capillary electrophoresis, inductively coupled plasma, atomic absorption spectroscopy, mass spectrometry, or any combination thereof.

[0075] The salt is recovered using at least one of the following techniques: filtration, precipitation with a non-solvent followed by filtration, evaporation of the solvent, or freeze-drying in the case of an aqueous solution.

[0076] The screening and characterization of pharmaceutically acceptable salts, solvates, and / or polymorphs can be accomplished using a variety of techniques, including but not limited to thermal analysis, X-ray diffraction, spectroscopy, microscopic methods, and elemental analysis. The various spectroscopic techniques used include but are not limited to Raman, FTIR, UVIS, and NMR (liquid and solid states). The various microscopic techniques include but are not limited to IR microscopy and Raman microscopy.

[0077] For the PI3Kδ inhibitors provided by the present invention, when the R5 substituent on the pyrazole group is a bulky group (such as haloalkyl or other branched alkyl, cycloalkylalkyl, heterocycloalkyl, etc.), compared with the compounds of the prior art, the inhibition of PI3Kδ phosphorylation is more significant both at the protein level and at the cellular level.

[0078] Pharmaceutical use of the present invention

[0079] The compound of formula (I) of the present invention, or its pharmaceutically acceptable salt, solvate, polymorph, ester, acid, isomer, metabolite, or prodrug, can inhibit the PI3Kδ kinase activity, so as to achieve the purpose of treating diseases or disorders related to PI3Kδ kinase activity.

[0080] Therefore, the present application relates to the use of the compound of formula (I) or its pharmaceutically acceptable salt, solvate, polymorph, ester, acid, isomer, metabolite, or prodrug in the preparation of a drug for inhibiting PI3Kδ kinase activity, or treating a disease or disorder related to PI3Kδ kinase activity.

[0081] The diseases or disorders are selected from chronic lymphocytic leukemia, non-Hodgkin lymphoma, Hodgkin lymphoma, mantle cell lymphoma, chronic obstructive pulmonary disease, rheumatoid arthritis, systemic lupus erythematosus, and asthma.

[0082] In an embodiment of the present invention, a medicament comprising a compound of the present invention can be administered to a patient by at least one of injection, oral administration, inhalation, rectal and transdermal administration. When treating a patient according to the present invention, the amount of the given medicament depends on many factors, such as the specific dosing regimen, the type and severity of the disease or disorder, and the uniqueness of the subject or host to be treated (such as body weight). However, depending on the specific circumstances, including for example the specific medicament employed, the route of administration, the disorder being treated, and the subject or host being treated, the dosage can be routinely determined by methods known in the art. Generally, for adult therapeutic use, the dosage is typically in the range of 0.02 - 5000 mg / day, for example about 1 - 1500 mg / day. The required dosage can conveniently be presented as a single dose, or as divided doses administered simultaneously (or within a short period of time) or at appropriate intervals, such as two, three, four or more divided doses per day. Those skilled in the art will understand that although the above dosage ranges are given, the specific effective amount can be appropriately adjusted according to the circumstances of the patient and in combination with the physician's diagnosis.

[0083] Preparation of the compound

[0084] The compounds of formula (I) can be synthesized using standard synthetic techniques known to those skilled in the art or using methods known in the art in combination with the methods described herein. Additionally, the solvents, temperatures and other reaction conditions given herein can be varied according to the art. As further guidance, the following synthetic methods can also be utilized.

[0085] The reactions can be used sequentially to provide the compounds described herein; or they can be used to synthesize fragments which are subsequently incorporated by the methods described herein and / or methods known in the art.

[0086] In certain embodiments, provided herein are methods for preparing and using the PI3Kδ inhibitor compounds described herein. In certain embodiments, the compounds described herein can be synthesized using the following synthetic schemes. Compounds can be synthesized using methods similar to those described below, by using appropriate alternative starting materials.

[0087] The starting materials for synthesizing the compounds described herein can be synthesized or can be obtained from commercial sources. The compounds described herein and other related compounds having different substituents can be synthesized using techniques and starting materials known to those skilled in the art. The general methods for preparing the compounds disclosed herein can be derived from reactions known in the art and the reaction can be modified by reagents and conditions considered appropriate by those skilled in the art to introduce the various moieties in the molecules provided herein.

[0088] If desired, the reaction products can be separated and purified using conventional techniques, including but not limited to filtration, distillation, crystallization, chromatography, and other methods. These products can be characterized using conventional methods, including physical constants and spectral data.

[0089] Preparation of Intermediates

[0090] Intermediate 1: Synthesis of 2-(1-chloropropyl)-3-cyclopropyl-5-fluoroquinazolin-4(3H)-one

[0091]

[0092] Step 1: To a solution of 2-amino-6-fluorobenzoic acid (5.0 g, 32.2 mmol, 1.0 eq) in tetrahydrofuran (50 mL) was added cyclopropylamine (2.21 g, 38.7 mmol, 1.2 eq), HATU (14.71 g, 38.7 mmol, 1.2 eq), and DIPEA (8.33 g, 64.5 mmol, 2.0 eq). The reaction was stirred at room temperature for 16 h. The solvent was removed under reduced pressure. The residue was diluted with ethyl acetate and the organic layer was washed successively with saturated brine and water, dried over anhydrous Na2SO4 for 30 min, filtered, concentrated, and triturated with petroleum ether and filtered to obtain an off-white solid. LC-MS (ESI): 195.1 (M+H) + 。

[0093] Step 2: At 0 °C, oxalyl chloride (COCl2) (5.40 g, 42.5 mmol, 1.2 eq) and a catalytic amount of DMF (0.06 mL) were added dropwise to a solution of 2-chlorobutyric acid (3.84 g, 35.42 mmol, 1.0 eq) in anhydrous tetrahydrofuran (40 mL). After the addition was complete, the reaction mixture was warmed to room temperature (25 °C) and stirred for 2 h, then used directly in the next step.

[0094] Step 3: At 0 °C, the intermediate obtained in Step 1 (32.2 mmol, 1.0 eq) was dissolved in anhydrous tetrahydrofuran (60 mL). DIPEA (12.49 g, 96.6 mmol, 3.0 eq) and the acyl chloride obtained in Step 2 (35.42 mmol, 1.1 eq) were added thereto. The reaction was stirred for 2 h. The reaction mixture was concentrated to dryness. The residue was diluted with dichloromethane and the organic phase was washed successively with saturated brine and water, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a solid compound. LC-MS (ESI): 285 (M+H) + 。

[0095] Step 4: Dissolve the intermediate obtained in Step 3 (32.2 mmol, 1.0 eq) in acetonitrile, add HMDS (25.82 g, 160 mmol, 5.0 eq)) and zinc chloride (21.81 g, 160 mmol, 5.0 eq) thereto. After stirring and reacting at 90 °C for 16 hours, evaporate the solvent under reduced pressure. Dilute the residue with dichloromethane, filter, concentrate the filtrate to dryness, and perform column chromatography (petroleum ether mixed solution of 3 - 6% ethyl acetate) to obtain 5.4 g of white solid compound Intermediate 1.

[0096] LC-MS(ESI): 281.7(M + H) + 。

[0097] The intermediates listed in Table 2 were synthesized by the above method:

[0098] Table 2

[0099]

[0100]

[0101]

[0102] Synthesis of Intermediate 14: 3-(1-(1-Ethoxyethyl)-1H-pyrazol-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine

[0103]

[0104] Step 1: Add a mixed solution of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (5.0 g, 19.2 mmol, 1.0 eq) to 1,4-dioxane / water (50 mL / 16 mL). Add 1-(1-ethoxyethyl)-4-pyrazolylboronic acid pinacol ester (7.01 g, 28.7 mmol, 1.5 eq) thereto. Replace the gas with nitrogen twice, add Pd(PPh3)4 (3.32 g, 2.87 mmol, 0.15 eq) and K2CO3 (5.29 g, 38.3 mmol, 2.0 eq) under a nitrogen atmosphere. After adding, replace the gas with nitrogen 5 times, and then raise the temperature to 135 °C and react for 24 hours. Concentrate and remove the solvent under reduced pressure. Add water (200 mL) to the residue, stir for 2 hours, filter, and wash the filter cake with dichloromethane twice to obtain an off-white solid compound Intermediate 14. LC-MS(ESI): 274.3(M + H) + 。

[0105] The intermediates listed in Table 3 were synthesized by the above method:

[0106] Table 3

[0107]

[0108]

[0109] Example 1.

[0110]

[0111] Dissolve Intermediate 1 (2 g, 7.5 mmol, 1.0 eq) in DMF (20 mL), add Intermediate 14 (2.26 g, 9.0 mmol, 1.2 eq) and K2CO3 (2.07 g, 15.0 mmol, 2.0 eq) thereto, and then raise the temperature to 60 °C and stir the reaction for 6 hours. Add water to the reaction solution, and a large amount of solid precipitates. After stirring for 15 minutes, filter, wash the filter cake 3 times with water, collect the filter cake, and separate by column chromatography (a mixed solution of dichloromethane containing 0 - 8% MeOH) to obtain 2.1 g of white solid Compound 1 in a yield of 57%. 1 H NMR (500 MHz, DMSO-d6) δ 8.28 - 8.22 (m, 2H), 7.80 - 7.75 (m, 2H), 7.45 (dd, J = 8.2, 1.0 Hz, 1H), 7.27 (ddt, J = 10.9, 8.2, 1.3 Hz, 1H), 6.49 (dd, J = 9.1, 5.0 Hz, 1H), 5.59 (q, J = 5.9 Hz, 1H), 3.49 - 3.39 (m, 1H), 3.27 (ddd, J = 7.1, 6.0, 3.5 Hz, 1H), 2.49 - 2.34 (m, 2H), 2.17 - 2.09 (m, 1H), 1.62 (d, J = 6.0 Hz, 3H), 1.21 - 1.08 (m, 3H), 1.04 (td, J = 7.0, 1.0 Hz, 3H), 0.89 (td, J = 7.3, 1.3 Hz, 3H), 0.81 - 0.74 (m, 1H).

[0112] Example 2.

[0113]

[0114] Referring to the synthesis method of Example 1, Compound 2 was synthesized from Intermediate 13 (2 g, 6.7 mmol, 1.0 eq) and Intermediate 21 (2.02 g, 8.03 mmol, 1.2 eq) to obtain 1.8 g of a white solid in a yield of 53%. 11H NMR (500 MHz, DMSO-d6) δ 8.51 (s, 1H), 8.26 (s, 1H), 8.00 (s, 1H), 7.95 - 7.57 (m, 3H), 7.10 (s, 2H), 6.52 (dd, J = 9.1, 5.0 Hz, 1H), 2.39 (ddd, J = 14.0, 9.0, 7.2 Hz, 1H), 2.16 (td, J = 6.9, 3.5 Hz, 1H), 1.26 - 1.07 (m, 3H), 0.91 (t, J = 7.3 Hz, 3H), 0.81 (tt, J = 9.9, 5.2 Hz, 1H). LC-MS (ESI): 513.2 (M + H) + .

[0115] Example 3.

[0116]

[0117] Referring to the synthesis method of Reference Example 1, Compound 3 was synthesized from Intermediate 13 (2 g, 6.7 mmol, 1.0 eq) and Intermediate 18 (1.95 g, 8.03 mmol, 1.2 eq) to obtain 1.5 g of a white solid with a yield of 44%. 1 1H NMR (500 MHz, DMSO-d6) δ 8.24 (s, 1H), 8.06 (s, 1H), 7.83 (ddd, J = 10.2, 9.0, 2.9 Hz, 1H), 7.68 (s, 1H), 7.65 - 7.53 (m, 1H), 6.50 (dd, J = 9.0, 5.0 Hz, 1H), 4.54 (p, J = 6.6 Hz, 1H), 2.37 (ddd, J = 13.9, 9.0, 7.2 Hz, 1H), 2.11 (dq, J = 9.6, 3.5, 2.7 Hz, 1H), 1.44 (dd, J = 6.6, 1.3 Hz, 6H), 1.26 - 1.06 (m, 4H), 0.89 (t, J = 7.3 Hz, 3H), 0.78 (ddd, J = 12.3, 6.3, 3.8 Hz, 1H). LC-MS (ESI): 505.2 (M + H) + .

[0118] Example 4.

[0119]

[0120] Referring to the synthesis method of Reference Example 1, Compound 4 was synthesized from Intermediate 1 (0.2 g, 0.71 mmol) and Intermediate 17 (0.22 g, 0.78 mmol) to obtain 79 mg of a white solid with a yield of 21%, 11H NMR (500 MHz, DMSO-d6) δ 8.24 (s, 1H), 8.11 (d, J = 0.7 Hz, 1H), 7.78 (td, J = 8.2, 5.4 Hz, 1H), 7.72 (d, J = 0.7 Hz, 1H), 7.45 (dd, J = 8.1, 1.0 Hz, 1H), 7.27 (ddd, J = 11.0, 8.2, 1.0 Hz, 1H), 6.47 (dd, J = 9.1, 5.0 Hz, 1H), 4.44 (td, J = 10.3, 5.0 Hz, 1H), 3.99 - 3.92 (m, 2H), 3.48 - 3.40 (m, 2H), 2.48 - 2.34 (m, 2H), 2.08 (td, J = 6.9, 3.4 Hz, 1H), 2.04 - 1.97 (m, 4H), 1.21 - 1.08 (m, 3H), 0.89 (t, J = 7.4 Hz, 3H), 0.80 - 0.73 (m, 1H).

[0121] Example 5.

[0122]

[0123] Referring to the synthesis method of Reference Example 1, compound 5 was synthesized from intermediate 1 (0.2 g, 0.71 mmol) and intermediate 18 (0.21 g, 0.85 mmol) to obtain 0.13 g of a white solid with a yield of 37%. 1 1H NMR (500 MHz, DMSO-d6) δ 8.24 (s, 1H), 8.07 (d, J = 0.8 Hz, 1H), 7.78 (td, J = 8.2, 5.5 Hz, 1H), 7.69 (d, J = 0.7 Hz, 1H), 7.45 (dd, J = 8.2, 1.0 Hz, 1H), 7.27 (ddd, J = 11.0, 8.2, 1.1 Hz, 1H), 6.47 (dd, J = 9.2, 5.0 Hz, 1H), 4.55 (p, J = 6.6 Hz, 1H), 2.47 - 2.35 (m, 2H), 2.12 - 2.06 (m, 1H), 1.44 (dd, J = 6.7, 1.4 Hz, 6H), 1.20 - 1.08 (m, 3H), 0.88 (t, J = 7.4 Hz, 3H), 0.77 (dp, J = 8.0, 5.2, 4.5 Hz, 1H).

[0124] Example 6.

[0125]

[0126] Referring to the synthesis method of Reference Example 1, compound 6 was synthesized from intermediate 1 (0.2 g, 0.71 mmol) and intermediate 19 (0.27 g, 0.85 mmol) as a white solid of 0.21 g with a yield of 50%. 1 H NMR (500 MHz, DMSO-d6) δ 8.25 (s, 1H), 8.12 (s, 1H), 7.78 (td, J = 8.2, 5.4 Hz, 1H), 7.67 (s, 1H), 7.45 (dd, J = 8.2, 1.0 Hz, 1H), 7.28 (dd, J = 10.9, 8.2 Hz, 1H), 6.47 (dd, J = 9.1, 5.0 Hz, 1H), 4.27 (t, J = 6.5 Hz, 2H), 3.51 (t, J = 4.6 Hz, 4H), 2.72 (t, J = 6.5 Hz, 2H), 2.48 - 2.32 (m, 6H), 2.06 (td, J = 6.9, 3.4 Hz, 1H), 1.13 (tdd, J = 17.0, 8.2, 5.5 Hz, 3H), 0.90 (t, J = 7.4 Hz, 3H), 0.77 (td, J = 8.4, 7.8, 4.2 Hz, 1H).

[0127] Example 7.

[0128]

[0129] Referring to the synthesis method of Reference Example 1, compound 7 was synthesized from intermediate 1 (0.2 g, 0.71 mmol) and intermediate 20 (0.21 g, 0.85 mmol) as a white solid of 89 mg with a yield of 25.6%. 1 H NMR (500 MHz, DMSO-d6) δ 8.25 (s, 1H), 8.12 (s, 1H), 7.78 (td, J = 8.2, 5.4 Hz, 1H), 7.67 (s, 1H), 7.45 (dd, J = 8.2, 1.0 Hz, 1H), 7.28 (dd, J = 10.9, 8.2 Hz, 1H), 6.47 (dd, J = 9.1, 5.0 Hz, 1H), 4.27 (t, J = 6.5 Hz, 2H), 3.51 (t, J = 4.6 Hz, 4H), 2.72 (t, J = 6.5 Hz, 2H), 2.48 - 2.32 (m, 6H), 2.06 (td, J = 6.9, 3.4 Hz, 1H), 1.13 (tdd, J = 17.0, 8.2, 5.5 Hz, 3H), 0.90 (t, J = 7.4 Hz, 3H), 0.77 (td, J = 8.4, 7.8, 4.2 Hz, 1H).

[0130] Example 8.

[0131]

[0132] Referring to the synthesis method of Reference Example 1, Compound 8 was synthesized from Intermediate 1 (0.2 g, 0.71 mmol) and Intermediate 21 (0.21 g, 0.85 mmol) to give 0.11 g of a white solid, with a yield of 32%. 1 H NMR (500 MHz, DMSO-d6) δ 8.52 (s, 1H), 8.26 (d, J = 5.6 Hz, 1H), 8.02 (s, 1H), 7.95 - 7.66 (m, 2H), 7.43 (d, J = 8.2 Hz, 1H), 7.27 (dd, J = 10.4, 8.3 Hz, 1H), 7.10 (s, 1H), 6.49 (dd, J = 9.1, 5.0 Hz, 1H), 2.49 - 2.43 (m, 1H), 2.43 - 2.33 (m, 1H), 2.20 - 2.10 (m, 1H), 1.22 - 1.16 (m, 1H), 1.16 - 1.06 (m, 2H), 0.90 (t, J = 7.3 Hz, 3H), 0.83 - 0.75 (m, 1H).

[0133] Example 9.

[0134]

[0135] Referring to the synthesis method of Reference Example 1, Compound 9 was synthesized from Intermediate 3 (0.2 g, 0.71 mmol) and Intermediate 18 (0.21 g, 0.85 mmol) to give 0.12 g of a white solid, with a yield of 36%. 1 H NMR (500 MHz, DMSO-d6) δ 8.24 (s, 1H), 8.06 (s, 1H), 7.80 - 7.62 (m, 4H), 6.98 (s, 1H), 6.50 (dd, J = 9.1, 5.0 Hz, 1H), 4.60 - 4.46 (m, 1H), 2.48 - 2.42 (m, 1H), 2.42 - 2.32 (m, 1H), 2.20 - 2.09 (m, 1H), 1.43 (dd, J = 6.6, 1.3 Hz, 6H), 1.26 - 1.07 (m, 3H), 0.88 (t, J = 7.3 Hz, 3H), 0.84 - 0.75 (m, 1H).

[0136] Example 10.

[0137]

[0138] Referring to the synthesis method of Reference Example 1, compound 10 was synthesized from intermediate 4 (0.2 g, 0.74 mmol) and intermediate 18 (0.22 g, 0.90 mmol) to give 0.14 g of a white solid with a yield of 42%. 1 H NMR (500 MHz, DMSO) δ 8.24 (s, 1H), 8.07 (s, 1H), 7.80 - 7.65 (m, 4H), 6.73 (s, 2H), 6.71 (t, J = 6.6 Hz, 1H), 4.61 - 4.49 (m, 1H), 2.19 - 2.09 (m, 1H), 1.84 (d, J = 6.6 Hz, 3H), 1.44 (d, J = 6.5 Hz, 6H), 1.26 - 1.15 (m, 1H), 1.14 - 1.03 (m, 2H), 0.80 (dd, J = 8.3, 3.4 Hz, 1H).

[0139] Example 11.

[0140]

[0141] Referring to the synthesis method of Reference Example 1, compound 11 was synthesized from intermediate 5 (0.2 g, 0.0.68 mmol) and intermediate 18 (0.2 g, 0.81 mmol) to give 0.16 g of a white solid with a yield of 48%. 1 H NMR (500 MHz, DMSO - d6) δ 8.31 (s, 1H), 8.09 (s, 1H), 7.83 (td, J = 8.2, 5.5 Hz, 1H), 7.70 (s, 1H), 7.55 (d, J = 8.1 Hz, 1H), 7.32 (dd, J = 10.6, 8.4 Hz, 1H), 7.00 (s, 1H), 6.10 (dd, J = 9.2, 5.1 Hz, 1H), 4.55 (dq, J = 13.3, 6.6 Hz, 1H), 4.08 (dd, J = 14.5, 5.6 Hz, 1H), 3.77 (dd, J = 14.5, 7.6 Hz, 1H), 2.46 (d, J = 8.9 Hz, 1H), 2.42 - 2.34 (m, 1H), 1.43 (dd, J = 6.6, 1.5 Hz, 6H), 0.75 (t, J = 7.3 Hz, 3H), 0.53 (s, 1H), 0.36 (tt, J = 10.3, 5.1 Hz, 2H), 0.18 - 0.06 (m, 1H), 0.02 (d, J = 4.9 Hz, 1H).

[0142] Example 12.

[0143]

[0144] Referring to the synthesis method of Reference Example 1, compound 12 was synthesized from intermediate 4 (0.2 g, 0.75 mmol) and intermediate 21 (0.23 g, 0.90 mmol), which was a white solid of 0.23 g with a yield of 64%. 1 H NMR (500 MHz, DMSO) δ 8.53 (s, 1H), 8.26 (s, 1H), 8.02 (s, 1H), 7.88 (d, J = 58.8 Hz, 1H), 7.78 - 7.68 (m, 3H), 7.14 (s, 2H), 6.74 (q, J = 6.6 Hz, 1H), 2.19 (t, J = 6.7 Hz, 1H), 1.87 (d, J = 6.6 Hz, 3H), 1.20 (dd, J = 9.3, 3.7 Hz, 1H), 1.10 (d, J = 5.4 Hz, 2H), 0.83 (dd, J = 8.4, 3.4 Hz, 1H).

[0145] Example 13.

[0146]

[0147] Referring to the synthesis method of Reference Example 1, compound 13 was synthesized from intermediate 3 (0.2 g, 0.71 mmol) and intermediate 21 (0.21 g, 0.85 mmol), which was a white solid of 0.13 g with a yield of 37%. 1 H NMR (500 MHz, DMSO) δ 8.52 (s, 1H), 8.26 (s, 1H), 8.01 (s, 1H), 7.87 (d, J = 58.8 Hz, 1H), 7.77 - 7.67 (m, 3H), 7.13 (s, 2H), 6.53 (d, J = 14.1 Hz, 1H), 2.48 (dd, J = 10.0, 4.8 Hz, 1H), 2.44 - 2.37 (m, 1H), 2.25 - 2.19 (m, 1H), 1.22 (dd, J = 11.5, 7.1 Hz, 1H), 1.19 - 1.10 (m, 2H), 0.91 (t, J = 7.3 Hz, 3H), 0.87 - 0.80 (m, 1H).

[0148] Example 14.

[0149]

[0150] Referring to the synthesis method of Reference Example 1, compound 14 was synthesized from intermediate 4 (0.2 g, 0.75 mmol) and intermediate 20 (0.22 g, 0.89 mmol), which was a white solid of 0.11 g with a yield of 32%. 11H NMR (500 MHz, DMSO-d6) δ 8.23 (s, 1H), 8.01 (s, 1H), 7.79 - 7.65 (m, 4H), 6.96 (s, 1H), 6.70 (q, J = 6.6 Hz, 1H), 4.89 (t, J = 5.6 Hz, 1H), 4.18 (t, J = 5.5 Hz, 2H), 3.74 (q, J = 5.6 Hz, 2H), 2.11 (tt, J = 7.2, 4.2 Hz, 1H), 1.84 (d, J = 6.6 Hz, 3H), 1.21 - 1.13 (m, 1H), 1.08 (q, J = 6.5 Hz, 2H), 0.79 (dt, J = 9.5, 4.1 Hz, 1H).

[0151] Example 15.

[0152]

[0153] Referring to the synthesis method of Example 1, compound 15 was synthesized from intermediate 3 (0.2 g, 0.71 mmol) and intermediate 20 (0.21 g, 0.85 mmol) to obtain 0.2 g of a white solid with a yield of 58%. 1 1H NMR (500 MHz, DMSO-d6) δ 8.24 (s, 1H), 8.01 (s, 1H), 7.79 - 7.62 (m, 4H), 6.99 (s, 1H), 6.50 (dd, J = 9.1, 5.0 Hz, 1H), 4.88 (t, J = 5.6 Hz, 1H), 4.18 (t, J = 5.5 Hz, 2H), 3.74 (q, J = 5.6 Hz, 2H), 2.48 - 2.43 (m, 1H), 2.38 (dt, J = 21.2, 7.3 Hz, 1H), 2.20 - 2.11 (m, 1H), 1.15 (dddd, J = 20.4, 16.7, 8.0, 4.9 Hz, 3H), 0.89 (t, J = 7.3 Hz, 3H), 0.82 - 0.75 (m, 1H).

[0154] Example 16.

[0155]

[0156] Referring to the synthesis method of Example 1, compound 16 was synthesized from intermediate 11 (0.2 g, 0.59 mmol) and intermediate 21 (0.18 g, 0.71 mmol) to obtain 0.18 g of a white solid with a yield of 33%. 11H NMR (500 MHz, DMSO-d6) δ 8.57 (d, J = 12.1 Hz, 1H), 8.06 (s, 1H), 8.03 - 7.90 (m, 2H), 7.86 - 7.57 (m, 2H), 7.53 - 7.37 (m, 1H), 7.28 - 6.97 (m, 2H), 6.88 (ddt, J = 22.0, 15.2, 8.3 Hz, 2H), 6.28 - 6.15 (m, 1H), 6.02 (dq, J = 33.9, 6.6 Hz, 1H), 1.74 (d, J = 6.5 Hz, 3H).

[0157] Example 17.

[0158]

[0159] Referring to the synthesis method of Example 1, compound 17 was synthesized from intermediate 10 (0.2 g, 0.76 mmol) and intermediate 21 (0.23 g, 0.91 mmol) to give 0.17 g of a white solid with a yield of 47%. 1 1H NMR (500 MHz, DMSO-d6) δ 8.52 (s, 1H), 8.25 (s, 1H), 8.01 (s, 1H), 7.81 (t, J = 58.7 Hz, 1H), 7.61 (t, J = 7.7 Hz, 1H), 7.42 (d, J = 8.0 Hz, 1H), 7.27 (d, J = 7.4 Hz, 1H), 7.12 (s, 1H), 6.69 (q, J = 6.6 Hz, 1H), 2.72 (s, 3H), 2.15 - 2.03 (m, 1H), 1.84 (d, J = 6.7 Hz, 3H), 1.17 - 1.11 (m, 1H), 1.10 - 1.01 (m, 2H), 0.74 (dd, J = 7.3, 3.2 Hz, 1H).

[0160] Example 18.

[0161]

[0162] Referring to the synthesis method of Example 1, compound 18 was synthesized from intermediate 10 (0.2 g, 0.76 mmol) and intermediate 18 (0.22 g, 0.91 mmol) to give 0.15 g of a white solid with a yield of 42%. 11H NMR (500 MHz, DMSO-d6) δ 8.24 (s, 1H), 8.07 (s, 1H), 7.68 (s, 1H), 7.62 (t, J = 7.8 Hz, 1H), 7.44 (d, J = 8.0 Hz, 1H), 7.27 (d, J = 7.4 Hz, 1H), 6.99 (s, 1H), 6.67 (q, J = 6.6 Hz, 1H), 4.54 (dt, J = 13.3, 6.7 Hz, 1H), 2.72 (s, 3H), 2.05 - 1.98 (m, 1H), 1.82 (d, J = 6.7 Hz, 3H), 1.44 (dd, J = 6.6, 1.2 Hz, 6H), 1.14 (dd, J = 8.6, 3.9 Hz, 1H), 1.09 - 0.99 (m, 2H), 0.75 - 0.64 (m, 1H).

[0163] Example 19.

[0164]

[0165] Referring to the synthesis method of Example 1, compound 19 was synthesized from intermediate 5 (0.2 g, 0.68 mmol) and intermediate 21 (0.2 g, 0.81 mmol) to give 0.15 g of a white solid, with a yield of 45%. 1 1H NMR (500 MHz, DMSO-d6) δ 8.54 (s, 1H), 8.33 (s, 1H), 8.03 (s, 1H), 7.93 - 7.66 (m, 2H), 7.54 (d, J = 8.2 Hz, 1H), 7.32 (dd, J = 10.9, 8.2 Hz, 1H), 7.26 - 7.05 (m, 1H), 6.13 (dd, J = 9.2, 5.2 Hz, 1H), 4.09 (dd, J = 14.5, 5.7 Hz, 1H), 3.81 (dd, J = 14.6, 7.5 Hz, 1H), 2.49 - 2.45 (m, 1H), 2.43 - 2.36 (m, 1H), 0.79 (t, J = 7.3 Hz, 3H), 0.59 (d, J = 7.2 Hz, 1H), 0.43 - 0.34 (m, 2H), 0.20 - 0.11 (m, 1H), 0.09 - 0.04 (m, 1H).

[0166] Example 20.

[0167]

[0168] Referring to the synthesis method of Example 1, compound 20 was synthesized from intermediate 5 (0.2 g, 0.68 mmol) and intermediate 20 (0.2 g, 0.81 mmol) to give 0.11 g of a white solid, with a yield of 33%. 11H NMR (500 MHz, DMSO-d6) δ 8.31 (s, 1H), 8.04 (d, J = 0.8 Hz, 1H), 7.83 (td, J = 8.2, 5.4 Hz, 1H), 7.70 (d, J = 0.8 Hz, 1H), 7.55 (dd, J = 8.2, 1.0 Hz, 1H), 7.36 - 7.28 (m, 1H), 6.10 (dd, J = 9.2, 5.2 Hz, 1H), 4.88 (t, J = 5.6 Hz, 1H), 4.18 (t, J = 5.5 Hz, 2H), 4.08 (dd, J = 14.6, 5.7 Hz, 1H), 3.84 - 3.77 (m, 1H), 3.74 (q, J = 5.6 Hz, 2H), 2.48 - 2.44 (m, 1H), 2.39 (ddd, J = 15.2, 7.9, 5.9 Hz, 1H), 0.77 (t, J = 7.3 Hz, 3H), 0.56 (d, J = 7.0 Hz, 1H), 0.41 - 0.33 (m, 2H), 0.13 (dq, J = 8.4, 2.7, 1.8 Hz, 1H), 0.06 - 0.00 (m, 1H).

[0169] Example 21.

[0170]

[0171] Referring to the synthesis method of Reference Example 1, compound 21 was synthesized from intermediate 6 (0.2 g, 0.68 mmol) and intermediate 18 (0.2 g, 0.81 mmol) to obtain 0.12 g of a white solid with a yield of 34%. 1 1H NMR (500 MHz, DMSO-d6) δ 8.32 (s, 1H), 8.08 (s, 1H), 7.86 - 7.80 (m, 2H), 7.75 (td, J = 8.7, 3.1 Hz, 1H), 7.69 (s, 1H), 6.13 (dd, J = 9.2, 5.2 Hz, 1H), 4.54 (p, J = 6.7 Hz, 1H), 4.14 (dd, J = 14.5, 5.7 Hz, 1H), 3.81 (dd, J = 14.5, 7.6 Hz, 1H), 1.43 (dd, J = 6.6, 2.0 Hz, 6H), 0.76 (t, J = 7.3 Hz, 3H), 0.55 (d, J = 7.6 Hz, 1H), 0.38 (ddd, J = 20.1, 9.4, 4.6 Hz, 2H), 0.13 (dq, J = 10.0, 4.6 Hz, 1H), 0.03 (dq, J = 10.0, 4.9 Hz, 1H).

[0172] Example 22.

[0173]

[0174] Referring to the synthesis method of Reference Example 1, Compound 22 was synthesized from Intermediate 12 (0.2 g, 0.65 mmol) and Intermediate 21 (0.2 g, 0.78 mmol) to give 0.10 g of a white solid with a yield of 31%. 1 H NMR (500 MHz, DMSO) δ 8.52 (s, 1H), 8.13 (s, 1H), 8.00 (s, 1H), 7.92 (dd, J = 9.3, 5.4 Hz, 1H), 7.86 - 7.62 (m, 3H), 7.31 (d, J = 1.9 Hz, 1H), 7.03 (s, 2H), 6.19 (q, J = 6.7 Hz, 1H), 5.71 (s, 1H), 3.59 (s, 3H), 1.74 (d, J = 6.7 Hz, 3H).

[0175] Example 23.

[0176]

[0177] Referring to the synthesis method of Reference Example 1, Compound 23 was synthesized from Intermediate 9 (0.2 g, 0.65 mmol) and Intermediate 21 (0.2 g, 0.78 mmol) to give 0.12 g of a white solid with a yield of 35%. 1 H NMR (500 MHz, DMSO) δ 8.54 (s, 1H), 8.11 (s, 1H), 8.02 (s, 1H), 7.92 - 7.86 (m, 1H), 7.84 - 7.66 (m, 3H), 7.49 (s, 1H), 7.00 (s, 3H), 6.11 (q, J = 6.6 Hz, 1H), 3.54 (s, 3H), 1.74 (d, J = 6.7 Hz, 3H).

[0178] Example 24.

[0179]

[0180] Referring to the synthesis method of Reference Example 1, Compound 24 was synthesized from Intermediate 8 (0.2 g, 0.70 mmol) and Intermediate 21 (0.21 g, 0.84 mmol) to give 0.14 g of a white solid with a yield of 40%. 11H NMR (500 MHz, DMSO) δ 8.52 (s, 1H), 8.26 (d, J = 5.5 Hz, 1H), 8.01 (s, 1H), 7.96 - 7.68 (m, 2H), 7.63 (dt, J = 16.2, 8.0 Hz, 1H), 7.13 (s, 2H), 6.73 (q, J = 6.6 Hz, 1H), 2.22 - 2.06 (m, 1H), 1.86 (d, J = 6.7 Hz, 3H), 1.28 - 1.18 (m, 1H), 1.13 - 1.04 (m, 2H), 0.81 (tdd, J = 16.9, 9.0, 3.5 Hz, 1H).

[0181] Example 25.

[0182]

[0183] Referring to the synthesis method of Reference Example 1, compound 25 was synthesized from intermediate 7 (0.2 g, 0.74 mmol) and intermediate 21 (0.22 g, 0.88 mmol) to give 0.17 g of a white solid with a yield of 52%. 1 1H NMR (500 MHz, DMSO-d6) δ 8.52 (s, 1H), 8.25 (s, 1H), 8.06 - 7.98 (m, 2H), 7.95 - 7.63 (m, 3H), 6.73 (q, J = 6.7 Hz, 1H), 2.18 (dq, J = 7.0, 3.5, 3.0 Hz, 1H), 1.86 (d, J = 6.6 Hz, 3H), 1.22 - 1.17 (m, 1H), 1.09 (d, J = 6.8 Hz, 2H), 0.83 (dt, J = 9.9, 4.7 Hz, 1H).

[0184] Example 26.

[0185]

[0186] Referring to the synthesis method of Reference Example 1, compound 26 was synthesized from intermediate 2 (1.0 g, 3.56 mmol) and intermediate 16 (1.13 g, 4.27 mmol) to give 1.2 g of a white solid with a yield of 66%. MS (ESI): [M+H] 508.9511.

[0187] Example 27.

[0188]

[0189] Referring to the synthesis method of Reference Example 1, compound 27 was synthesized from intermediate 6 (0.2 g, 0.68 mmol) and intermediate 22 (0.22 g, 0.82 mmol) to give 0.13 g of a white solid with a yield of 38%.1 1H NMR (500 MHz, CDCl3) δ 8.40 (s, 1H), 7.89 (dd, J = 8.5, 2.9 Hz, 1H), 7.84 - 7.77 (m, 2H), 7.70 (s, 1H), 7.46 (td, J = 8.7, 2.9 Hz, 1H), 6.13 (dd, J = 8.6, 6.1 Hz, 1H), 5.60 (s, 2H), 4.29 (dd, J = 14.5, 6.1 Hz, 1H), 4.00 - 3.84 (m, 2H), 2.73 - 2.46 (m, 2H), 1.97 - 1.79 (m, 4H), 1.09 - 0.99 (m, 1H), 0.94 (t, J = 7.3 Hz, 3H), 0.80 (dd, J = 12.8, 7.3 Hz, 6H), 0.67 - 0.53 (m, 2H), 0.49 - 0.34 (m, 2H).

[0190] Example 28.

[0191]

[0192] Referring to the synthesis method of Reference Example 1, compound 28 was synthesized from intermediate 6 (0.2 g, 0.68 mmol) and intermediate 15 (0.2 g, 0.8 mmol) to give 0.12 g of a white solid, with a yield of 34%. 1 1H NMR (500 MHz, CDCl3) δ 8.42 (s, 1H), 7.94 - 7.85 (m, 3H), 7.79 (dd, J = 8.9, 4.9 Hz, 1H), 7.46 (td, J = 8.6, 3.0 Hz, 1H), 6.13 (dd, J = 8.6, 6.1 Hz, 1H), 5.59 (s, 2H), 5.51 - 5.44 (m, 1H), 5.12 - 5.02 (m, 4H), 4.29 (dd, J = 14.5, 6.1 Hz, 1H), 3.90 (dd, J = 14.6, 7.3 Hz, 1H), 2.58 (ddt, J = 35.8, 13.9, 7.0 Hz, 2H), 1.06 - 0.98 (m, 1H), 0.94 (t, J = 7.3 Hz, 3H), 0.66 - 0.52 (m, 2H), 0.49 - 0.36 (m, 2H).

[0193] Example 29.

[0194]

[0195] Referring to the synthesis method of Reference Example 1, compound 29 was synthesized from intermediate 6 (0.2 g, 0.68 mmol) and intermediate 24 (0.21 g, 0.82 mmol) to give 0.17 g of a white solid, with a yield of 50%.1 1H NMR (500 MHz, CDCl3) δ 8.40 (s, 1H), 7.88 (dd, J = 8.5, 2.8 Hz, 1H), 7.83 (s, 1H), 7.79 (dd, J = 9.0, 4.9 Hz, 1H), 7.77 (s, 1H), 7.45 (td, J = 8.7, 2.9 Hz, 1H), 6.12 (dd, J = 8.5, 6.1 Hz, 1H), 5.78 (s, 2H), 4.29 (dd, J = 14.5, 6.0 Hz, 1H), 4.01 (d, J = 7.0 Hz, 2H), 3.88 (dd, J = 14.6, 7.3 Hz, 1H), 2.67 - 2.49 (m, 2H), 1.34 - 1.28 (m, 1H), 1.05 - 0.97 (m, 1H), 0.94 (t, J = 7.3 Hz, 3H), 0.67 (d, J = 7.3 Hz, 2H), 0.64 - 0.53 (m, 2H), 0.47 - 0.42 (m, 1H), 0.41 - 0.35 (m, 3H).

[0196] Example 30.

[0197]

[0198] Referring to the synthesis method of Reference Example 1, compound 30 was synthesized from intermediate 5 (0.2 g, 0.68 mmol) and intermediate 15 (0.21 g, 0.81 mmol) to obtain 0.12 g of a white solid with a yield of 34%. 1 1H NMR (500 MHz, CDCl3) δ 8.41 (s, 1H), 7.92 (s, 1H), 7.88 (s, 1H), 7.65 (dd, J = 14.8, 6.8 Hz, 1H), 7.57 (d, J = 8.1 Hz, 1H), 7.16 - 7.07 (m, 1H), 6.11 (dd, J = 8.6, 6.0 Hz, 1H), 5.63 (s, 2H), 5.52 - 5.45 (m, 1H), 5.13 - 5.02 (m, 4H), 4.26 (dd, J = 14.6, 6.1 Hz, 1H), 3.85 (dd, J = 14.6, 7.3 Hz, 1H), 2.71 - 2.45 (m, 2H), 1.06 - 0.98 (m, 1H), 0.94 (t, J = 7.3 Hz, 3H), 0.64 - 0.53 (m, 2H), 0.42 (dtt, J = 15.4, 10.4, 5.2 Hz, 2H).

[0199] Example 31.

[0200]

[0201] Referring to the synthesis method of Reference Example 1, compound 31 was synthesized from intermediate 5 (0.2 g, 0.68 mmol) and intermediate 22 (0.22 g, 0.82 mmol) as a white solid (0.13 g) with a yield of 39%. 1 H NMR (500 MHz, CDCl3) δ 8.40 (s, 1H), 7.80 (s, 1H), 7.71 (s, 1H), 7.66 (td, J = 8.1, 5.3 Hz, 1H), 7.59 (d, J = 7.9 Hz, 1H), 7.11 (dd, J = 10.0, 8.6 Hz, 1H), 6.10 (dd, J = 8.7, 6.0 Hz, 1H), 5.62 (s, 2H), 4.26 (dd, J = 14.6, 6.1 Hz, 1H), 3.99 - 3.90 (m, 1H), 3.84 (dd, J = 14.6, 7.4 Hz, 1H), 2.72 - 2.46 (m, 2H), 1.96 - 1.81 (m, 4H), 1.08 - 0.98 (m, 1H), 0.95 (t, J = 7.4 Hz, 3H), 0.81 (td, J = 7.3, 4.8 Hz, 6H), 0.64 - 0.53 (m, 2H), 0.47 - 0.36 (m, 2H).

[0202] Example 32.

[0203]

[0204] Referring to the synthesis method of Reference Example 1, compound 32 was synthesized from intermediate 5 (0.2 g, 0.68 mmol) and intermediate 23 (0.21 g, 0.82 mmol) as a white solid (0.13 g) with a yield of 38%. 1 H NMR (500 MHz, CDCl3) δ 8.39 (s, 1H), 7.78 (s, 1H), 7.70 (s, 1H), 7.65 (td, J = 8.1, 5.4 Hz, 1H), 7.57 (d, J = 8.0 Hz, 1H), 7.11 (dd, J = 9.8, 8.4 Hz, 1H), 6.09 (dd, J = 8.6, 6.0 Hz, 1H), 5.71 (s, 2H), 4.25 (dd, J = 14.6, 6.1 Hz, 1H), 4.03 - 3.88 (m, 2H), 3.84 (dd, J = 14.6, 7.3 Hz, 1H), 2.70 - 2.47 (m, 2H), 2.24 (dp, J = 13.6, 6.8 Hz, 1H), 1.05 - 0.97 (m, 1H), 0.96 - 0.88 (m, 9H), 0.63 - 0.52 (m, 2H), 0.47 - 0.35 (m, 2H).

[0205] Example 33.

[0206]

[0207] Referring to the synthesis method of Reference Example 1, compound 33 was synthesized from intermediate 6 (0.2 g, 0.68 mmol) and intermediate 23 (0.21 g, 0.82 mmol) to give 0.15 g of a white solid with a yield of 50%. 1 H NMR (500 MHz, CDCl3) δ 8.40 (s, 1H), 7.88 (dd, J = 8.5, 2.8 Hz, 1H), 7.80 (dd, J = 8.9, 4.9 Hz, 1H), 7.77 (s, 1H), 7.69 (s, 1H), 7.45 (td, J = 8.7, 2.9 Hz, 1H), 6.12 (dd, J = 8.5, 6.1 Hz, 1H), 5.69 (s, 2H), 4.29 (dd, J = 14.5, 6.1 Hz, 1H), 3.94 (dd, J = 7.2, 4.2 Hz, 2H), 3.89 (dd, J = 14.6, 7.3 Hz, 1H), 2.70 - 2.47 (m, 2H), 2.23 (dp, J = 13.6, 6.8 Hz, 1H), 1.07 - 0.97 (m, 1H), 0.97 - 0.88 (m, 9H), 0.64 - 0.53 (m, 2H), 0.41 (ddq, J = 25.4, 10.2, 4.9 Hz, 2H).

[0208] Example 34.

[0209]

[0210] Referring to the synthesis method of Reference Example 1, compound 34 was synthesized from intermediate 3 (0.2 g, 0.71 mmol) and intermediate 15 (0.22 g, 0.85 mmol) to give 0.14 g of a white solid with a yield of 36%. 1 H NMR (500 MHz, CDCl3) δ 8.36 (s, 1H), 7.87 (d, J = 1.1 Hz, 2H), 7.82 (dd, J = 8.4, 2.8 Hz, 1H), 7.70 (dd, J = 8.9, 4.9 Hz, 1H), 7.42 (td, J = 8.7, 2.9 Hz, 1H), 6.62 (dd, J = 8.4, 6.2 Hz, 1H), 5.72 (s, 2H), 5.54 - 5.41 (m, 1H), 5.16 - 4.99 (m, 4H), 2.65 - 2.46 (m, 2H), 2.45 - 2.34 (m, 1H), 1.52 - 1.41 (m, 1H), 1.34 - 1.22 (m, 2H), 1.01 (t, J = 7.3 Hz, 3H), 0.92 (ddd, J = 21.3, 13.2, 5.5 Hz, 1H).

[0211] Example 35.

[0212]

[0213] Referring to the synthesis method of Reference Example 1, Compound 35 was synthesized from Intermediate 3 (0.2 g, 0.71 mmol) and Intermediate 23 (0.22 g, 0.85 mmol) to give 0.150 g of a white solid, with a yield of 47%. 1 H NMR (500 MHz, DMSO-d6) δ 8.24 (s, 1H), 8.06 (s, 1H), 7.80 - 7.63 (m, 4H), 6.98 (s, 1H), 6.50 (dd, J = 9.1, 5.0 Hz, 1H), 3.95 (d, J = 7.2 Hz, 2H), 2.46 (dd, J = 9.9, 4.7 Hz, 1H), 2.42 - 2.34 (m, 1H), 2.19 - 2.09 (m, 2H), 1.22 - 1.06 (m, 3H), 0.89 (t, J = 7.4 Hz, 3H), 0.85 (d, J = 6.7 Hz, 6H), 0.79 (dt, J = 9.6, 3.9 Hz, 1H).

[0214] Example 36.

[0215]

[0216] Referring to the synthesis method of Reference Example 1, Compound 36 was synthesized from Intermediate 4 (0.2 g, 0.74 mmol) and Intermediate 23 (0.23 g, 0.88 mmol) to give 0.14 g of a white solid, with a yield of 42%. 1 H NMR (500 MHz, DMSO-d6) δ 8.23 (s, 1H), 8.06 (s, 1H), 7.88 - 7.60 (m, 4H), 6.98 (s, 1H), 6.70 (q, J = 6.6 Hz, 1H), 3.95 (d, J = 7.2 Hz, 2H), 2.19 - 2.06 (m, 2H), 1.84 (d, J = 6.7 Hz, 3H), 1.21 - 1.14 (m, 1H), 1.13 - 1.03 (m, 2H), 0.85 (d, J = 6.7 Hz, 6H), 0.82 - 0.75 (m, 1H).

[0217] Example 37.

[0218]

[0219] Referring to the synthesis method of Reference Example 1, compound 37 was synthesized from intermediate 4 (0.2 g, 0.74 mmol) and intermediate 22 (0.24 g, 0.88 mmol), which was a white solid of 0.12 g with a yield of 35%. 1 H NMR (500 MHz, DMSO-d6) δ 8.24 (s, 1H), 8.12 (s, 1H), 7.79 - 7.64 (m, 4H), 6.71 (q, J = 6.6 Hz, 1H), 4.16 - 3.94 (m, 1H), 2.17 (qd, J = 7.4, 3.7 Hz, 1H), 1.84 (d, J = 6.6 Hz, 3H), 1.83 - 1.71 (m, 4H), 1.22 - 1.14 (m, 1H), 1.13 - 1.05 (m, 2H), 0.84 - 0.78 (m, 1H), 0.70 (t, J = 7.3 Hz, 6H).

[0220] Example 38.

[0221]

[0222] Referring to the synthesis method of Reference Example 1, compound 38 was synthesized from intermediate 3 (0.2 g, 0.71 mmol) and intermediate 22 (0.23 g, 0.88 mmol), which was a white solid of 0.14 g with a yield of 40%. 1 H NMR (500 MHz, DMSO-d6) δ 8.25 (s, 1H), 8.12 (s, 1H), 7.86 - 7.58 (m, 4H), 6.88 (s, 1H), 6.51 (dd, J = 9.1, 5.0 Hz, 1H), 4.16 - 3.96 (m, 1H), 2.48 - 2.43 (m, 1H), 2.38 (dt, J = 21.2, 7.4 Hz, 1H), 2.19 (ddd, J = 11.5, 6.9, 4.4 Hz, 1H), 1.89 - 1.70 (m, 4H), 1.22 - 1.18 (m, 1H), 1.18 - 1.09 (m, 2H), 0.88 (t, J = 7.3 Hz, 3H), 0.83 - 0.77 (m, 1H), 0.70 (t, J = 7.3 Hz, 6H).

[0223] Example 39.

[0224]

[0225] Referring to the synthesis method of Reference Example 1, compound 39 was synthesized from intermediate 4 (0.2 g, 0.74 mmol) and intermediate 24 (0.22 g, 0.88 mmol), which was a white solid of 0.15 g with a yield of 44%. 11H NMR (500 MHz, DMSO-d6) δ 8.24 (s, 1H), 8.10 (s, 1H), 7.80 - 7.64 (m, 4H), 7.07 (dd, J = 106.0, 54.9 Hz, 1H), 6.71 (q, J = 6.6 Hz, 1H), 4.00 (d, J = 7.2 Hz, 2H), 2.16 - 2.08 (m, 1H), 1.84 (d, J = 6.7 Hz, 3H), 1.31 - 1.26 (m, 1H), 1.19 (dd, J = 10.4, 4.3 Hz, 1H), 1.12 - 1.03 (m, 2H), 0.82 - 0.73 (m, 1H), 0.56 - 0.47 (m, 2H), 0.38 (q, J = 4.9 Hz, 2H).

[0226] Example 40.

[0227]

[0228] Referring to the synthesis method of Reference Example 1, Compound 40 was synthesized from Intermediate 4 (0.2 g, 0.75 mmol) and Intermediate 15 (0.23 g, 0.9 mmol) to obtain 0.14 g of a white solid with a yield of 38%. 1 1H NMR (500 MHz, DMSO-d6) δ 8.24 (s, 1H), 8.10 (s, 1H), 7.80 - 7.64 (m, 4H), 7.07 (dd, J = 106.0, 54.9 Hz, 1H), 6.71 (q, J = 6.6 Hz, 1H), 4.00 (d, J = 7.2 Hz, 2H), 2.16 - 2.08 (m, 1H), 1.84 (d, J = 6.7 Hz, 3H), 1.31 - 1.26 (m, 1H), 1.19 (dd, J = 10.4, 4.3 Hz, 1H), 1.12 - 1.03 (m, 2H), 0.82 - 0.73 (m, 1H), 0.56 - 0.47 (m, 2H), 0.38 (q, J = 4.9 Hz, 2H).

[0229] Comparative Example Compound 1.

[0230]

[0231] Comparative Example Compound 1 was prepared by referring to the synthesis method of Reference Example 1.

[0232] Example 41. In vitro inhibitory activity (enzyme activity) assay

[0233] The in vitro enzyme activity experiment was used to determine the IC of the compound against the type I kinase of the PI3K family (PI3Kδ, stock solution concentration 5.9 mM) 50Values. Protein kinase PI3Kδ was purchased from Invitrogen (USA); substrates PIP2:3PS, PI, and ADP-Glo kit were purchased from Promega (USA). 5.4 μL of protein kinase PI3Kδ diluted to a certain concentration (final concentration 1 ng / μL) with the protein diluent in the kit was taken and mixed with 1 μL of each compound diluted in gradient (diluted with the reaction buffer in the kit as the solvent). Incubate at room temperature for 1 hour (final concentrations of the compounds were 10 μM, 1 μM, 0.3 μM, 0.1 μM, 0.03 μM, 0.01 μM, 0.003 μM, 0.001 μM); add the substrate (final concentration of the substrate 0.01 mg / mL) and mix. React the mixed system at room temperature for 1 hour; finally, add 8 μL of the detection reagent, incubate at 25 °C for 40 minutes, and read the fluorescence value using an MD SpectraMax I3X microplate reader (Molecular Devices, USA). Based on the read fluorescence values, plot the graph using Prism 8.0 (GraphPad Software, San Diego, CA), calculate the IC50 values of the compounds 12 and 24 of the present invention, comparative example compound 1, and TGR-1202 (purchased from MCE, China) for the tested protein kinase PI3Kδ, as shown in Table 4 below.

[0234] Table 4

[0235] <![CDATA[IC 50 :nM]]> <![CDATA[IC 50 :nM]]> <![CDATA[IC 50 :nM]]> TGR-1202 48.4 Compound 10 6.85 Compound 19 18.1 Compound 2 2.15 Compound 11 10.99 Compound 20 22.6 Compound 3 3.15 Compound 12 2.59 Compound 21 15.4 Compound 5 14.5 Compound 13 14.1 Compound 24 2.12 Compound 7 15.4 Compound 16 3.96 Compound 39 24 Compound 8 16.8 Compound 17 3.72 Compound 40 23.7 Compound 9 13.2 Compound 18 3.63 Comparative example compound 1 34

[0236] Experiments show that the compounds of the present invention have strong inhibitory activity against PI3Kδ. Especially when the substituents on the pyrazole ring are replaced with larger steric hindrance groups (such as haloalkyl or other branched alkyl groups, etc.), their inhibitory activity against PI3Kδ is significantly improved.

[0237] Example 42. Detection of PI3Kδ inhibitory activity in cells

[0238] This example was carried out using the human lymphoma cell line Jeko-1 cells with high expression of PI3Kδ (purchased from Nanjing Kebai Biotechnology Co., Ltd.). Specifically: Seed the cells in a 6-well plate, 5×10 5Cells were plated in 6-well plates at a density of 5×10^5 cells per well. Different concentrations of the compound of the present invention (dissolved in DMSO) were added to the 6-well plates, and the final concentrations of the compound in the reaction system were 0.03 μM, 0.1 μM, 0.3 μM, 1 μM, 3 μM, and 10 μM, respectively. Then, the cells were incubated in an incubator for 2 hours. The cells were collected, proteins were extracted, and the phosphorylation level of the T473 site of AKT (a direct downstream protein of PI3Kδ) in each group was detected by western blot (the antibody used was purchased from CST, Cat: 4060S, USA). GAPDH was used as an internal reference, and then quantification was performed using Image J software, and plotting was done using Graphad 8.0 to calculate the EC 50 value, as shown in Table 5.

[0239] Table 5

[0240] Compound <![CDATA[EC 50 :nM]]> Compound <![CDATA[EC 50 :nM]]> Compound <![CDATA[EC 50 :nM]]> Comparative example compound 1 460 Compound 9 80 Compound 13 100 Compound 5 83 Compound 10 80 Compound 14 150 Compound 7 85 Compound 11 97 Compound 15 128 Compound 8 85 Compound 12 80 Compound 24 46

[0241] The experimental results showed that the compound of the present application had a more significant inhibitory effect on PI3Kδ phosphorylation compared to Comparative Example Compound 1.

[0242] Example 43. Detection of Inhibitory Activity of PI3Kα, PI3Kβ, and PI3Kγ in Cells

[0243] In this example, human breast cancer cell line SK-Br-3 cells with high expression of PI3Kα (purchased from Nanjing Kebai Biotechnology Co., Ltd.), human breast cancer cells MDA-MB-468 with high expression of PI3Kβ (purchased from Nanjing Kebai Biotechnology Co., Ltd.), and mouse lymphoma cells RAW264.4 with high expression of PI3Kγ (purchased from Nanjing Kebai Biotechnology Co., Ltd.) were used. Specifically: Cells were plated in 6-well plates at a density of 5×10 5 ^5 cells per well. Different concentrations of the compound of the present invention (dissolved in DMSO) were added to the 6-well plates, and the final concentrations of the compound in the reaction system were 0.03 μM, 0.1 μM, 0.3 μM, 1 μM, 3 μM, and 10 μM, respectively. Then, the cells were incubated in an incubator for 2 hours. The cells were collected, proteins were extracted, and the phosphorylation level of the T473 site of AKT (a direct downstream protein of different subtypes of PI3K) in each group was detected by western blot (the antibody used was purchased from CST, Cat: 4060S, USA). GAPDH was used as an internal reference, and then quantification was performed using Image J software, and plotting was done using Graphad 8.0 to calculate the EC 50 value, as shown in Table 6.

[0244] Table 6

[0245]

[0246] The experimental results show that the EC50 values of the compounds of this application for the phosphorylation of other PI3K subtypes such as PI3Kα, PI3Kβ, and PI3Kγ are all > 1000 nM, indicating that the compounds of this application have no inhibitory effect on the phosphorylation of other PI3K subtypes, while having a strong inhibitory effect on the phosphorylation of PI3Kδ, proving that the compounds of this invention can selectively inhibit the activity of PI3Kδ.

[0247] Example 44. In vivo pharmacodynamic detection on a mouse xenograft tumor model of DOHH2 cells

[0248] In this example, the experimental results of Compound 12, Compound 24, and the control compound Linperlisib (purchased from MedChemExpress, China) in a mouse xenograft tumor model of DOHH2 cells (purchased from ATCC) were respectively tested.

[0249] The experimental steps are as follows:

[0250] (1) Six-week-old male C57BL / 6J mice were purchased from Jiangsu Jicui Yakang Biotech Co., Ltd. and raised in an SPF-class laboratory. The drinking water and bedding were both sterilized by high-pressure steam. All operations related to the mice were carried out under sterile conditions.

[0251] (2) On day 0, approximately 1×10 6 DOHH2 cells (purchased from ATCC) were subcutaneously injected into the left back of all mice.

[0252] (3) Starting from day 6, the mice in each group were orally administered daily: a vehicle containing 5% (v / v) DMSO, 10% (v / v) propylene glycol (purchased from Xi'an Tianzheng Pharmaceutical Excipients, China), and 10% (v / v) HS-15 (polyethylene glycol-15-hydroxystearate, purchased from BASF, Germany) once a day (5 mice); Compound 12 at a dose of 50 mg / kg body weight once a day (5 mice each); Compound 24 at a dose of 50 mg / kg body weight once a day (5 mice each); Linperlisib at a dose of 50 mg / kg body weight once a day (5 mice). The oral administration volume of each group was equal.

[0253] (4) Starting from day 0, the length / width of the subcutaneous tumor was measured daily with a vernier caliper, and the body weight of the mice was recorded daily to determine the effects of Compound 12, Compound 24, and the control compound Linperlisib on the body weight of the mice.

[0254] (5) The growth trend of the subcutaneous tumor was statistically analyzed. The tumor volume was calculated as: length × width × width / 2 mm 3 .

[0255] The experimental results are as Figure 1and 2 As shown. Compounds 12, 24 and the control compound Linperlisib showed good effects in inhibiting tumors in a mouse xenograft tumor model. Moreover, with the increase in the number of days of drug administration, the inhibitory effects of Compounds 12 and 24 on mouse tumors became more significant, and were superior to those of the control compound Linperlisib.

[0256] Example 45. In Vivo Efficacy Detection on a Mouse Xenograft Tumor Model of MC38 Cells

[0257] In this example, the efficacy of Comparative Compounds 1 and 24 was tested and compared in a mouse model of MC38 cells.

[0258] The experimental steps are as follows:

[0259] (1) C57BL / 6J male mice at 6 weeks old for use in a MC38 cell xenograft tumor model were purchased from Jiangsu Genscript Biotech Corporation. All the above mice were housed in an SPF-class laboratory, and the drinking water and bedding were sterilized by high-pressure steaming. All operations related to the mice were carried out under sterile conditions.

[0260] (2) On day 0, approximately 1×10 6 MC38 cells (purchased from ATCC) were subcutaneously injected into the left dorsal side of C57BL / 6J mice.

[0261] (3) Starting from day 0, the mice were randomly divided into three groups of 5 mice each and began to be administered drugs for 22 days. Among them, the first group of mice was orally administered a solvent containing only 10% (v / v) HS-15 with the same volume as the drug administration group; Comparative Compounds 1 and 24 with a dose of 30 mg / kg body weight were respectively administered to the second and third groups of mice once a day.

[0262] (4) Starting from day 0, the body weights of the mice were recorded daily to determine the effects of Compound 24 and Comparative Compound 1 on the body weights of the mice. The results are shown in Figure 3 .

[0263] (5) Starting from day 0, the length and width of the subcutaneous tumors were measured daily with vernier calipers, and the growth trends of subcutaneous tumors in each group of mice were statistically analyzed. The calculation method for tumor volume: length × width × width / 2 mm 3 , and the results are shown in Figure 4 .

[0264] (6) After 22 days of drug administration, the mice were euthanized, and the tumors were removed and weighed. The results are shown in Figure 5 .

[0265] The results of this example are shown in Figure 3 - Figure 5 . Among them, Figure 3Shows the change in the average body weight of mice in different treatment groups (shown as relative body weight in the figure: percentage calculated based on the weight of the mice at the start of drug administration) over time in the MC38 cell mouse xenograft tumor model; Figure 4 Shows the change in the average size of tumors (shown as the absolute size of the tumor in the figure) in different treatment groups over time in the MC38 cell mouse xenograft tumor model; Figure 5 Shows the average tumor weight and the calculated tumor growth inhibition rate (TGI) of mice in different treatment groups on the 14th day after drug administration in the MC38 cell mouse xenograft tumor model.

[0266] Figure 4 The experimental results show that in the MC38 cell mouse xenograft tumor model, the group of compound 24 showed a very good effect of inhibiting mouse tumors. Figure 5 The experimental results show that for the group of compound 24, the tumor growth inhibition rate on the 22nd day after drug administration in the MC38 cell mouse xenograft tumor model was as high as 86.7% (see Figure 5 ), where the tumor growth inhibition rate (TGI) = (weight of tumors in the control group - weight of tumors in the experimental group) / weight of tumors in the control group. This shows that the compound 24 of the present invention can significantly inhibit the growth of tumors in the MC38 cell animal model. Additionally, Figure 3 the results also show that compound 24 not only effectively inhibits the growth of mouse tumors, but also has basically no effect on the body weight of mice, indicating that compound 24 is suitable for animal administration.

[0267] The present invention provides a PI3Kδ inhibitor, which can be used to treat subjects with diseases related to the activity of PI3Kδ. Thus, the present invention is suitable for industrial applications.

[0268] Although the present invention has been described in detail herein, the present invention is not limited thereto, and those skilled in the art of this technology can make modifications based on the principles of the present invention. Therefore, all modifications made in accordance with the principles of the present invention should be understood to fall within the protection scope of the present invention.

Claims

1. A PI3Kδ inhibitor, comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof: Wherein, R1 are each independently selected from halogen and C1-C4 alkyl, and m is 1 or 2; R2 is selected from C3-C6 cycloalkyl, C3-C6 cycloalkyl C1-C4 alkyl, and phenyl or pyrazolyl optionally substituted by halogen or methyl; R3 and R4 are each independently selected from H and C1-C4 alkyl; R5 is selected from C3-C8 branched alkyl, C1-C8 haloalkyl, C1-C4 alkoxy C1-C4 alkyl, C2-C6 hydroxyalkyl, C3-C6 cycloalkyl C1-C4 alkyl, 4-6 membered heterocycloalkyl, and 4-6 membered heterocycloalkyl C1-C4 alkyl.

2. The PI3Kδ inhibitor according to claim 1, wherein, R5 is selected from isopropyl, isobutyl, pent-3-yl, monohaloalkyl, dihaloalkyl, 1-ethoxy-ethyl, hydroxyethyl, cyclopropylmethyl, oxetanyl, tetrahydropyranyl or morpholinylethyl.

3. The PI3Kδ inhibitor according to claim 1, wherein, R5 is haloethyl or dihalomethyl.

4. The PI3Kδ inhibitor according to claim 1, wherein, R5 is difluoromethyl.

5. The PI3Kδ inhibitor according to any one of claims 1-4, wherein, R1 is selected from fluorine and methyl.

6. The PI3Kδ inhibitor according to any one of claims 1-4, wherein R2 is selected from cyclopropyl, cyclopropylmethyl, fluorophenyl, and N-methyl-pyrazolyl.

7. The PI3Kδ inhibitor according to any one of claims 1-4, wherein, One of R3 and R4 is H, and the other is methyl or ethyl.

8. The PI3Kδ inhibitor according to any one of claims 1-4, which comprises the following compound or a pharmaceutically acceptable salt thereof:

9. A pharmaceutical composition, which comprises the PI3Kδ inhibitor according to any one of claims 1-8, and a pharmaceutically acceptable carrier or excipient, and optionally other active ingredients.

10. Use of the PI3Kδ inhibitor according to any one of claims 1-8 in the preparation of a drug for inhibiting PI3Kδ kinase activity or treating a disease or disorder associated with PI3Kδ kinase activity.

11. The use according to claim 10, wherein, The disease or disorder is selected from chronic lymphocytic leukemia, non-Hodgkin lymphoma, Hodgkin lymphoma, mantle cell lymphoma, chronic obstructive pulmonary disease, rheumatoid arthritis, systemic lupus erythematosus and asthma.

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