WO2018082444A1 - Pyrazolopyrimidine compound as pi3k inhibitor and use thereof - Google Patents

Abstract

The present invention relates to the pyrazolopyrimidine compound of formula (I) and a pharmaceutically-acceptable salt thereof. Such compounds can be used to inhibit the activity of a fat kinase PI3K, and can also be used to treat diseases mediated by PI3K, such as cancers, inflammation and autoimmune diseases. The present invention also relates to a pharmaceutical composition containing such compounds, a method for preparing such compounds and the use of such compounds or pharmaceutical composition in preparing drugs for treating cancers, inflammation and autoimmune diseases mediated by PI3K.

Description

Pyrazolopyrimidine compound as PI3K inhibitor and its application Technical field

The invention belongs to the field of chemical pharmacy, and particularly relates to a class of PI3K inhibitors and pharmaceutical compositions thereof, and a preparation method and application thereof.

Background technique

Phosphatidylinositol 3-kinases (PI3Ks) are ubiquitous adipokines that act both as signal transducers downstream of cell surface receptors and as signal transducers in the constituent intracellular membrane and protein trafficking pathways (Vanhaesebroeck, B. et al., Nature Rev. Mol. Cell Biol., 2010, 11, 329-341). The PI3K family of adipokines can be divided into three groups according to the specificity of their physiological substrates: Type I, Type II and Type III. Among these three types, Type I has been extensively studied. Type I PI3K is a heterodimer composed of a p110 catalyzed subunit and a regulated subunit. Type I PI3Ks are further divided into Type Ia and Type Ib enzymes. Type Ia enzymes are composed of three different catalytic subunits (p110, p110 and p110) dimerized with five different regulatory subunits (p85, p55, p50, p85 and p55), all of which are catalytic subunits It is capable of interacting with all regulatory subunits to form a variety of heterodimers known as PI3K, PI3K and PI3K. P110 and p110 are expressed essentially in all cell types, while p110 is predominantly expressed in leukocytes. A single type of Ib enzyme consists of a p110 catalytic subunit that interacts with the p101 regulatory subunit and is referred to as PI3K. Like p110, the type Ib enzyme is mainly expressed in white blood cells.

PI3Ks play a role in tumorigenesis in many types of cancer, which is caused by dysregulation or overactivation of the PI3K/AKT pathway (Vivanco and Sawyers, Nature Rev. Cancer, 2002, 2, 489-501). Among the four subtypes, PI3K controls B- in certain B-cell cancers. The survival of the cells plays a role. For example, PI3K is important for the survival of non-Hodgkin's lymphoma (NHL) and chronic lymphocytic leukemia (CLL). The PI3K inhibitor idelalisib has been clinically proven to treat CLL (Furman, RR, et al., The New Englang Jounal of Medicine, 2014, 370, 997-1007; O'Brien, S., et al., Blood, 2015, 126, 2686-2694), and approved by the US FDA for the treatment of these diseases. This fully demonstrates that inhibition of PI3K activity can treat B-cell lymphoma and leukemia, including NHL and CLL.

In addition, inhibition of PI3K can disrupt regulatory T cell-mediated immune tolerance to tumors and increase immune response, leading to tumor regression in animal models (Ali, K. et al., Nature, 2014, 510, 407). -411). These findings suggest that inhibition of PI3K sum is valuable for the treatment of tumors, especially those with insufficient immune response, such as breast cancer, lung cancer (including small cell lung cancer, non-small cell lung cancer, bronchoalveolar carcinoma), prostate cancer. , cholangiocarcinoma, bone cancer, bladder cancer, head and neck cancer, kidney cancer, liver cancer, gastrointestinal cancer, esophageal cancer, ovarian cancer, pancreatic cancer, skin cancer, testicular cancer, thyroid cancer, uterine cancer, cervical cancer and vagina Cancer, leukemia, multiple myeloma and lymphoma.

In addition to tumors, there is evidence that PI3K also plays an important role in inflammation and autoimmune diseases (Puri, KD et al., J. Immunol., 2009, 182 (Supp 1.50), 14; Maxwell, MJ Et al., J. Autoimmunity, 2012, 38, 381-391; Suarez-Fueyo, A. et al., J. Immunol., 2011, 187, 2376-2385). Thus, inhibition of PI3K can be used to treat inflammatory and autoimmune diseases including, but not limited to, skin inflammation, rheumatoid arthritis, allergic rhinitis, asthma, Crohn's disease, chronic obstructive pulmonary disease (COPD), systems Lupus erythematosus, psoriasis, multiple sclerosis, activated PI3K syndrome, and Sjogren syndrome.

The present invention relates to a new generation of PI3K inhibitors useful for the treatment of cancer, inflammatory and autoimmune diseases.

Summary of the invention

In one aspect of the invention, there is provided a compound of formula (I):

Or a pharmaceutically acceptable salt thereof, wherein:

R ^{1 is} selected from H, halogen, cyano, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl, wherein said alkyl, alkenyl, alkynyl, An aryl, heteroaryl, cycloalkyl, or heterocycloalkyl group is optionally substituted with from 1 to 5 R ^1a ;

R ^{1a is} selected from the group consisting of H, anthracene, halogen, cyano, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl, wherein said alkyl, alkenyl, alkyne Alkyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl optionally substituted by halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , alkyl, alkenyl, alkynyl, cycloalkyl , or a heterocycloalkyl group;

R ^{2 is} selected from aryl, heteroaryl, cycloalkyl, or heterocycloalkyl, wherein R ^{2 is} optionally substituted with from 1 to 5 R ^2a ;

R ^{2a is} selected from the group consisting of H, halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , NR ^b C(O)R ^d , NR ^b C(O)OR ^d , NR ^b C(O)NR ^b R ^c , NR ^b S(O) ₂ R ^d , NR ^b S(O) ₂ NR ^b R ^c , C(O)NR ^b R ^c , C(O)OR ^d , S(O) ₂ NR ^b R ^c , C (O) R ^d , S(O) ₂ R ^d , P(O)R ^b R ^c , alkyl, alkenyl, or alkynyl;

R ^{3 is} selected from the group consisting of H, anthracene, alkyl, alkenyl, alkynyl, haloalkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cyanoalkyl, cycloalkyl, or cycloalkylalkyl ;

R ^{4 is} selected from the group consisting of H, alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, cyanoalkyl, cycloalkyl, or cycloalkylalkyl;

R ^{5 is} selected from H, halogen, cyano, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl, wherein said alkyl, alkenyl, alkynyl, An aryl group, a heteroaryl group, a cycloalkyl group, or a heterocycloalkyl group may be substituted with from 1 to 3 R ^5a ;

R ^{5a is} selected from the group consisting of H, hydrazine, halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , NR ^b C(O)R ^d , NR ^b C(O)OR ^d , NR ^b C(O)NR ^b R ^c , NR ^b S(O) ₂ R ^d , NR ^b S(O) ₂ NR ^b R ^c , C(O)NR ^b R ^c , C(O)OR ^d , S(O) ₂ NR ^b R ^c , C(O)R ^d , S(O) ₂ R ^d , P(O)R ^b R ^c , alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl ;

R ⁶ and R ^{7 are} each selected from H, an amino group, or an alkyl group;

R ^a , R ^b , R ^c , and R ^{d are} independently selected from the group consisting of H, alkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cyanoalkyl, alkenyl, alkynyl, cycloalkyl, heterocycle An alkyl group, an aryl group, or a heteroaryl group;

Or R ^b and R ^c together with the nitrogen atom to which they are attached form a 4 to 7 membered heterocycloalkyl group, optionally substituted with 1 to 3 R ^e ;

R ^{e is} selected from the group consisting of H, halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , NR ^b C(O)R ^d , NR ^b C(O)OR ^d , NR ^b C(O)NR ^b R ^c , NR ^b S(O) ₂ R ^d , NR ^b S(O) ₂ NR ^b R ^c , C(O)NR ^b R ^c , C(O)OR ^d , S(O) ₂ NR ^b R ^c , C (O) R ^d , S(O) ₂ R ^d , P(O)R ^b R ^c , alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl.

Preferably, the invention provides a compound of formula (II):

Or a pharmaceutically acceptable salt thereof, wherein:

R ^{1 is} selected from H, halogen, cyano, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl, wherein said alkyl, alkenyl, alkynyl, An aryl, heteroaryl, cycloalkyl, or heterocycloalkyl group is optionally substituted with from 1 to 5 R ^1a ;

R ^{1a is} selected from the group consisting of H, anthracene, halogen, cyano, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl, wherein said alkyl, alkenyl, alkyne Alkyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl optionally substituted by halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , alkyl, alkenyl, alkynyl, cycloalkyl , or a heterocycloalkyl group;

R ^{2a is} selected from the group consisting of H, halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , NR ^b C(O)R ^d , NR ^b C(O)OR ^d , NR ^b C(O)NR ^b R ^c , NR ^b S(O) ₂ R ^d , NR ^b S(O) ₂ NR ^b R ^c , C(O)NR ^b R ^c , C(O)OR ^d , S(O) ₂ NR ^b R ^c , C (O) R ^d , S(O) ₂ R ^d , P(O)R ^b R ^c , alkyl, alkenyl, or alkynyl;

R ^{3 is} selected from the group consisting of H, anthracene, alkyl, alkenyl, alkynyl, haloalkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cyanoalkyl, cycloalkyl, or cycloalkylalkyl ;

R ⁶ and R ^{7 are} each selected from H, an amino group, or an alkyl group;

R ^a , R ^b , R ^c , and R ^{d are} independently selected from the group consisting of H, alkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cyanoalkyl, alkenyl, alkynyl, cycloalkyl, heterocycle An alkyl group, an aryl group, or a heteroaryl group;

Or R ^b and R ^c together with the nitrogen atom to which they are attached form a 4 to 7 membered heterocycloalkyl group, optionally substituted with 1 to 3 R ^e ;

R ^{e is} selected from the group consisting of H, halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , NR ^b C(O)R ^d , NR ^b C(O)OR ^d , NR ^b C(O)NR ^b R ^c , NR ^b S(O) ₂ R ^d , NR ^b S(O) ₂ NR ^b R ^c , C(O)NR ^b R ^c , C(O)OR ^d , S(O) ₂ NR ^b R ^c , C (O) R ^d , S(O) ₂ R ^d , P(O)R ^b R ^c , alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;

n is 1, 2, or 3.

In a preferred embodiment, R ^{1 is} selected from the group consisting of halogen, cyano, alkyl, haloalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl.

In a more preferred embodiment, R ^{1 is} selected from halogen.

In a preferred embodiment, R ^{2a is} selected from the group consisting of H and halogen.

In a more preferred embodiment, R ^{2a is} selected from the group consisting of H and fluorine.

In a preferred embodiment, R ^{3 is} selected from the group consisting of alkyl, haloalkyl, and cycloalkyl.

In a more preferred embodiment, R ^{3 is} selected from an alkyl group.

In a preferred embodiment, R ^{4 is} selected from the group consisting of H and alkyl.

In a more preferred embodiment, R ^{4 is} selected from H.

In a preferred embodiment, R ^{5 is} selected from the group consisting of halogen, cyano, alkynyl, cycloalkylalkynyl, arylalkynyl, heteroarylalkynyl, aryl, and heteroaryl.

In a more preferred embodiment, R ^{5 is} selected from cyano.

In a preferred embodiment, R ^{6 is} selected from the group consisting of H, amino, and alkyl.

In a preferred embodiment, R ^{7 is} selected from the group consisting of H and amino.

The term "halo" or "halogen" includes fluoro, chloro, bromo, and iodo.

The term "alkyl" refers to a straight or branched saturated hydrocarbon group. Examples of the alkyl group include methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, tert-butyl), pentyl ( For example, n-pentyl, isopentyl, neopentyl), hexyl (eg n-hexyl, 2-hexyl, 3-hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl) , 3-ethylpentyl-1, etc., heptyl (eg n-heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 2-methylhexyl, 3-methylhexyl, 2, 2-dimethylpentyl, 3,3-dimethylpentyl, 3-ethylpentyl-1, etc.), octyl (eg 1-octyl, 2-octyl, 2-ethylhexyl, etc.) , fluorenyl (such as 1-indenyl), fluorenyl (such as n-decyl), and the like. In particular, it is a linear or branched alkyl group having 1 to 20 carbon atoms, more specifically a linear or branched alkyl group having 1 to 10 atoms, and more preferably 1 to 6 atoms. A linear or branched alkyl group. Unless otherwise defined, all radical definitions in the invention are as defined herein.

The term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein the alkyl group is as defined above.

The term "haloalkyl" refers to an alkyl group having one or more halo substituents. The alkyl group and the halo or halogen are as defined above. Examples of haloalkyl groups include CH ₂ F, CHF ₂ , CF ₃ , C ₂ F ₅ , CCl ₃ , and the like.

The term "cyanoalkyl" refers to an alkyl group substituted with a cyano group (-CN).

The term "alkenyl" refers to a hydrocarbyl group having one or more C=C double bonds. Examples of alkenyl groups include ethenyl, propenyl, allyl, 1-butenyl, 2-butenyl, 1,3-butadienyl, 1-pentenyl, 2-pentenyl, 1,3-pentadienyl, 1-hexenyl, 2-hexenyl and the like, and the like.

The term "alkynyl" refers to a hydrocarbyl group having one or more C≡C triple bonds. Examples of alkynyl groups include ethynyl, propynyl, propargyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 1-hexynyl, 2- An alkynyl group, etc., and the like.

The term "cycloalkyl" refers to a non-aromatic carbocyclic ring including a cyclized alkyl group, a cyclized alkenyl group, and a cyclized alkynyl group. The cycloalkyl group can be a monocyclic or polycyclic ring system (e.g., having 2, 3 or 4 fused rings), including spiro rings. In certain embodiments, a cycloalkyl group can have from 3 to 20 carbon atoms. The cycloalkyl group may further have 0, 1, 2 or 3 C=C double bonds and/or 0, 1 or 2 C≡C triple bonds. Also included in the definition of cycloalkyl are those having one or more aromatic rings fused to a cycloalkyl ring (for example, having a common bond), such as pentane, pentene, hexane, Benzo derivatives of alkenes, and the like, and similar compounds. The cycloalkyl group having one or more fused aromatic rings may be bonded through an aromatic moiety or a non-aromatic moiety. Examples of the cycloalkyl group include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, a cyclopentenyl group, a cyclohexenyl group, a cyclohexadienyl group, a cycloheptenyl group, and a cycloheptyl group. Dienyl, adamantyl, and the like.

The term "heterocycloalkyl" refers to a non-aromatic heterocyclic ring wherein one or more of the atoms forming the ring are heteroatoms such as O, N, S or phosphorus. The heterocyclyl group may include a monocyclic or polycyclic ring (such as a ring system having 2, 3 or 4 fused rings) and a spiro ring. Examples of preferred "heterocycloalkyl" groups include, but are not limited to, 吖: aziridinyl (aziridine), azetidinyl (azetidinyl), tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, oxazolidinyl, thiazolidinyl, imidazolidinyl, Isoxazolylalkyl, isothiazolidinyl, pyrazolidinyl, morpholinyl, thiomorpholinyl, piperazinyl, piperidinyl, and the like. Also included in the definition of heterocycloalkyl are those having one or more aromatic rings fused to a non-aromatic heterocycloalkyl ring (for example, having a shared bond), such as 2,3-dihydrobenzene. And furyl, 1,3-benzodioxolyl, benzo-1,4-dioxanyl, phthalimido, naphthalimide, and the like Group. Heterocycloalkyl groups having one or more fused aromatic rings may be attached through an aromatic moiety or a non-aromatic moiety. The nitrogen and sulfur atoms in the heterocycloalkyl group may exist in an oxidized form.

The term "aryl" refers to a monocyclic or polycyclic (eg, having 2, 3 or 4 fused rings) aromatic hydrocarbons such as phenyl, naphthyl, anthracenyl, phenanthryl, anthracenyl, and the like.

The term "heteroaryl" refers to an aromatic heterocycle having at least one heteroatom ring member such as O, N or S. Heteroaryl groups include monocyclic or polycyclic (eg, having 2, 3 or 4 fused rings) ring systems. Any N atom ringd in the heterocyclic group can also be oxidized to form an N-oxide. Examples of preferred "heteroaryl" groups include, but are not limited to, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, furyl, thienyl, imidazolyl, triazolyl, tetrazole , thiazolyl, isothiazolyl, 1,2,4-thiadiazolyl, pyrrolyl, pyrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, benzofuranyl, benzothienyl ,benzothiazolyl, indenyl, oxazolyl, quinolyl, isoquinolyl, fluorenyl, oxazolyl, benzimidazolyl, pyrrolopyridyl, pyrrolopyrimidinyl, pyrazolopyridinyl , pyrazolopyrimidinyl, and the like.

The term "compound", as used herein, is meant to include all stereoisomers, geometric isomers, tautomers, isotopes.

The compounds of the invention may be asymmetric, for example having one or more stereocenters. Unless otherwise defined, all stereoisomers are, for example, enantiomers and diastereomers. Compounds of the invention containing asymmetrically substituted carbon atoms can be separated into optically pure or racemic forms. Optically pure forms can be prepared by resolution of the racemate or by the use of chiral synthons or chiral reagents.

The compounds of the invention may also include tautomeric forms. The new form of tautomer is produced by the interchange of a single bond and an adjacent double bond with proton transfer.

The compounds of the invention may also include all isotopic forms of the atoms present in the intermediate or final compound. Isotopes include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include deuterium and tritium.

The invention also includes pharmaceutically acceptable salts of the compounds of formula (I). By pharmaceutically acceptable salt is meant a derivative of a compound wherein the parent compound is modified by conversion of the base moiety present to its salt form, or wherein the parent compound is modified by conversion of the acid moiety present to its salt form. a derivative of the compound. Examples of pharmaceutically acceptable salts include, but are not limited to, salts of inorganic or organic acids of basic groups such as amines, or salts of inorganic or organic bases of acidic groups such as carboxylic acids. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound of formula I by reacting the free base form of these compounds with from 1 to 4 equivalents of the appropriate acid in a solvent system. Suitable salts are in ^{Remington's Pharmaceutical Sciences, 17 th ed} ., Mack Publishing Company, Easton, Pa., 1985, p.1418 , and Journal of Pharmaceutical Science, are listed in (1977) 66, 2.

The compounds of the present invention, as well as pharmaceutically acceptable salts thereof, also include solvated or hydrated forms. In general, solvated or hydrated forms are equivalent to the unsolvated or non-hydrated forms and are included within the scope of the invention. Some of the compounds of the invention may exist in a variety of crystalline forms or in amorphous form. In general, all physical forms of the compounds are included within the scope of the invention.

The invention also includes prodrugs of the compounds of formula (I). A prodrug is a pharmacological substance (ie, a drug) derived from a parent drug that is metabolized into a parent drug in the body once administered. Prodrugs can be prepared by substituting one or more functional groups present in the compound, wherein the substitution in the prodrug The base is removed in vivo in such a way as to convert to the parent compound. The preparation and use of prodrugs can be found in T. Higuchi and V. Stella, "Pro-drugs as Novel Delivery Systems," Vol. 14 of the ACSSymposium Series and Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American. Found in the Pharmaceutical Association and Pergamon Press, 1987. .

The invention also provides a composition comprising a compound of formula (I) together with a pharmaceutically acceptable carrier or excipient. The composition of the present invention can be administered orally, parenterally (injectable), spray inhalation, topical administration, rectal administration, nasal administration, vaginal administration, intraperitoneal administration or via implantation. Into the reservoir for administration.

In another aspect of the invention, the invention provides a method of modulating kinase activity using a compound of formula (I). The term "modulating kinase activity" as used herein, refers to reduced kinase activity when contacted with a compound of formula (I) of the invention, relative to kinase activity in the absence of a compound of formula (I). Accordingly, the present application provides methods of modulating kinase activity by contacting a kinase with a compound of formula (I) of the invention.

In some embodiments, the kinase is a lipokinase, such as PI3K, PI3K, PI3K, or PI3K.

In some embodiments, the kinase is PI3K.

In another aspect of the invention, the invention provides a method of the compounds of the invention for use in the treatment of a disease mediated by PI3K. Diseases associated with PI3K include cancer, inflammation and autoimmune diseases.

In some embodiments, the cancer of the invention is a solid tumor, leukemia, lymphoma, and multiple myeloma.

In some embodiments, the leukemia of the invention is selected from the group consisting of acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), and chronic myelogenous leukemia (CML); Lymphoma is selected from Hodgkin's lymphoma, non-Hodgkin's lymphoma (NHL), and thin Lymphoma (MCL), follicular lymphoma, B-cell lymphoma, T-cell lymphoma, and diffuse large B-cell lymphoma.

In some embodiments, the autoimmune disease and inflammatory disease of the present invention are selected from the group consisting of skin inflammation, rheumatoid arthritis, allergic rhinitis, asthma, Crohn's disease, chronic obstructive pulmonary disease (COPD), system Lupus erythematosus, psoriasis, multiple sclerosis, activated PI3K syndrome, and Sjogren syndrome.

In another aspect of the invention, the invention provides the use of a compound of the invention, or a pharmaceutically acceptable salt or pharmaceutical composition thereof, in combination with one or more drugs for the treatment of cancer, inflammation and autoimmune diseases.

In some embodiments, the combination medicaments referred to in the present invention include small molecule compounds and macromolecular antibody drugs. Small molecule compounds are selected from, but are not limited to, various kinase inhibitors such as BTK inhibitors, as well as other small molecule non-kinase inhibitors, etc.; macromolecular drugs include anti-CD20, anti-CTLA4, anti-PD-1, anti-PD- L1 antibody and the like.

In another aspect of the invention, the invention provides a process for the preparation of a compound of formula (I). The compounds of the present invention can be prepared by the reaction procedures and methods described below.

Reaction process 1

Compounds of formula (I) can be prepared using the synthetic routes outlined in Reaction Scheme 1. Starting material I-1 and N,N-dimethylformamide dimethyl acetal were refluxed in toluene to give I-2. Compound I-2 is reacted with 3-aminopyrazole in acetic acid to give the ring-closing product pyrazolopyrimidine I-3. The chlorination of I-3 with NCS gave the chlorinated product I-4. After hydrolysis of the ester group in I-4, the formed carboxylic acid I-5 is condensed with N,O-dimethylhydroxylamine to give the amide compound I-6. After the addition of the amide bond in the format reagent R ³ MgCl and I-6, the formed ketone I-7 is condensed with the (R)-sulfinamide to give I-8. The imino group in I-8 is reduced with lithium tri-sec-ylborohydride to give the sulfinamide compound I-9 in the S configuration. After the sulfinyl group is removed with hydrochloric acid, the free amino group I-10 is reacted with YL (Y is a R ⁵ , R ⁶ , and R ⁷ -substituted pyrimidine, and L is a leaving group) to give the final product (I).

Specific embodiment

The following examples are intended to illustrate preferred embodiments of the invention. It will be apparent to those skilled in the art that the technology disclosed in the following examples represents a technique discovered by the inventors and which functions well in the practice of the invention, and thus may be considered as a preferred form of its implementation. However, it will be apparent to those skilled in the <Desc/Clms Page number .

Example 1.

(S)-4-Amino-6-(1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethylamine)pyrimidine-5-carbonitrile

Step 1. 5-Phenylpyrazole [1,5-a]pyrimidine-6-carboxylic acid ethyl ester

In a 250 mL round bottom flask, ethyl benzoylformate (10 g, 56.2 mmol), N,N-dimethylformamide dimethyl acetal (6.7 g, 56.2 mmol), toluene 120 mL, refluxed for 2 hours. . After the reaction was completed, toluene was distilled off under reduced pressure. The obtained residue was dissolved in 200 mL of ethanol, and 3-aminopyrazole (4.7 g, 56.6 mmol) and 40 mL of glacial acetic acid were added under stirring, and the mixture was reacted at room temperature for 8 hours. After completion of the reaction, ethanol was evaporated under reduced pressure, and the residue was diluted with H ₂ O, and extracted with CH ₂ Cl ₂ . The obtained organic phase was washed successively with a saturated Na ₂ CO ₃ solution, saturated NaCI solution, dried over anhydrous Na ₂ SO ₄ , filtered, and then evaporated. LCMS (ESI): m / z = 268 (M + H) +.

Step 2. Ethyl 3-chloro-5-phenylpyrazole [1,5-a]pyrimidine-6-carboxylate

Ethyl 5-phenylpyrazole [1,5-a]pyrimidine-6-carboxylate (14.2 g, 53.1 mmol) was dissolved in 150 mL of CHCl ₃ and NCS (7.1 g, 53.2 mmol) was added and the mixture was warmed to 60 ° C 6 hours. After completion of the reaction, it was diluted with CH ₂ Cl ₂ and washed successively with 0.1N aqueous HCl and saturated NaCI. The organic phase was dried over anhydrous Na ₂ SO _4, filtered and concentrated to give 15.5 g of a yellow solid, was used directly in the next step, 97% yield. LCMS (ESI): m / z = 302 (M + H) +.

Step 3. 3-Chloro-5-phenylpyrazole [1,5-a]pyrimidine-6-carboxylic acid

In a 250 mL round bottom flask, 3-chloro-5-phenylpyrazole [1,5-a]pyrimidine-6-carboxylic acid ethyl ester (15.5 g, 51.5 mmol), methanol 50 mL, 2N NaOH aqueous solution 100 mL, room temperature Reaction for 5 hours. After completion of the reaction, methanol was distilled off under reduced pressure, filtered, and the residue was washed with H ₂ O. The obtained filtrate was adjusted to pH = 3 with 4N aqueous HCl, and solid was precipitated and filtered to give 8.2 g of a solid. LCMS (ESI): m / z = 274 (M + H) +.

Step 4. 3-Chloro-N-methoxy-N-methyl-5-phenylpyrazole [1,5-a]pyrimidine-6-amide

3-Chloro-5-phenylpyrazole [1,5-a]pyrimidine-6-carboxylic acid (8.2 g, 30 mmol), DIEA (11.6 g, 90 mmol) was dissolved in 100 mL CH ₂ Cl ₂ and stirred. N,O-Dimethylhydroxylamine hydrochloride (3.2 g, 33 mmol), HATU (12.5 g, 33 mmol). After completion of the reaction, diluted with CH ₂ Cl _2, the organic phase was washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered and concentrated, residue was purified by column obtained (PE:EtOAc (v / v) = 2:1) A solid of 8.5 g was obtained with a yield of 90%. LCMS (ESI): m / z = 317 (M + H) +.

Step 5. 1-(3-Chloro-5-phenylpyrazole [1,5-a]pyrimidin-6-yl)ethanone

3-Chloro-N-methoxy-N-methyl-5-phenylpyrazole [1,5-a]pyrimidin-6-amide (8.5 g, 26.9 mmol) was dissolved in 80 mL of THF under N2. Cool in an ice bath. The solution was slowly added dropwise to a solution of methylmagnesium chloride in THF (3M, 14 mL). The temperature was maintained for 1 hour after the completion of the dropping. After completion of the reaction, the reaction was quenched by carefully adding a saturated NH ₄ Cl solution, and THF was evaporated under reduced pressure. The aqueous phase was extracted with EtOAc, the combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, and concentrated to give 6.5 g of a solid, 89% yield after filtration. LCMS (ESI): m / z = 272 (M + H) +.

Step 6. (R,E)-N-(1-(3-Chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethylidene)-2-methylpropane-2 - sulfenamide

(R)-tert-butylsulfinamide (3 g, 24.8 mmol) was dissolved in 30 mL of THF, and 1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl was added dropwise. Ethyl ketone (6.5 g, 24 mmol) and tetraethyl titanate (11 g, 48.2 mmol) in THF (50 mL). Then, the temperature was refluxed for 4 hours. After completion of the reaction, it was cooled to room temperature, a saturated NH ₄ Cl solution was added, and THF was evaporated under reduced pressure. The aqueous phase was extracted with EtOAc, the combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered and concentrated, the resulting residue was purified by column chromatography (PE:EtOAc (v / v) = 1:1) to give a solid 4.2 g, yield 45%. LCMS (ESI): m / z = 375 (M + H) +.

Step 7. (R)-N-((S)-1-(3-Chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethyl)-2-methylpropane- 2-sulfinamide

(R,E)-N-(1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethylidene)-2-methylpropane under nitrogen The -2-sulfinamide (4.2 g, 11.2 mmol) was dissolved in 40 mL of THF and cooled to -40. While maintaining the temperature, a solution of lithium tri-sec-butylborohydride in THF (1 M, 22 mL) was slowly added dropwise. The temperature was maintained for 1 hour after the completion of the dropping. After completion of the reaction, saturated NH ₄ Cl solution, stirred for 5min, warmed to room temperature, evaporated under reduced pressure to remove THF, water phase was extracted with EtOAc, the combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered Concentration, the residue was purified by EtOAc EtOAcjjjjjjj LCMS (ESI): m / z = 377 (M + H) +.

Step 8. (S)-1-(3-Chloro-5-phenylpyrazole [1,5-a]pyrimidin-6-yl)ethylamine

(R)-N-((S)-1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethyl)-2-methylpropan-2- The sulfinamide (2 g, 5.3 mmol) was dissolved in 10 mL of MeOH, and concentrated hydrochloric acid (4 mL). After completion of the reaction, it was cooled in an ice bath and adjusted to pH = 9 with a saturated Na ₂ CO ₃ solution. Extracted with EtOAc phase was distilled under reduced pressure to remove methanol, water, and the combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, and concentrated to give a solid 1.2g, 83% yield after filtration. LCMS (ESI): m / z = 273 (M + H) +.

Step 9. (S)-4-Amino-6-(1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethylamine)pyrimidine-5-carbonitrile

In a 25 mL round bottom flask, (S)-1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethylamine (150 mg, 0.55 mmol), 4-amino -5-Nitrile-6-chloropyrimidine (85 mg, 0.55 mmol), KF (64 mg, 1.1 mmol), DIEA (355 mg, 2.75 mmol), DMSO (5 mL), and warmed to 90 ° C for 2 hours. After completion of the reaction, diluted with H ₂ O, EtOAc. The combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered and concentrated. The residue was purified by column chromatography (EtOAc:EtOAc:EtOAc LCMS (ESI): m / z = 391 (M + H) +. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.69 (s, 1H), 8.03 (s, 1H), 8. s (s, 1H), 7.84-7.69 (m, 1H), 7.66-7.55 (m, 3H), 7.52-7.32 (m, 1H), 5.59 (d, J = 6.2 Hz, 1H), 5.51 (s, 2H), 5.26-5.14 (m, 1H), 1.57 (d, J = 7.1 Hz, 3H).

Example 2.

(S)-2,4-diamino-6-(1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethylamine)pyrimidine-5-carbonitrile

Step 1. 2,4,6-Trichloropyrimidine-5-formaldehyde

In 150mL round bottom flask, was added POCl ₃ (76.5g, 0.5mmol), cooled in an ice bath, was slowly added DMF (5.7g, 78mmol), maintaining the temperature of the reaction 20min. To the solution was added barbituric acid (10 g, 78 mmol), the ice bath was removed, and the reaction was heated to reflux for 12 hours. After completion of the reaction, distillation under reduced pressure to remove excess POCl _3, the residue poured onto crushed ice, extracted with EtOAc, the combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, and concentrated to give 10g oil filtered , used directly in the next step of the reaction. The yield was 61%.

Step 2. 2,4,6-Trichloropyrimidine-5-carbonitrile

In 150mL round bottom flask was added 2,4,6-trichloro-pyrimidine-5-carbaldehyde (10g, 47.4mmol), hydroxylamine hydrochloride (3.3g, 47.5mmol), glacial acetic acid (100 mL) and H ₂ O (5mL) Heat to 60 ° C for 1 hour. After completion of the reaction, the reaction solution was poured into crushed ice, extracted in CH ₂ Cl _2, the combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered and concentrated, the resultant was dissolved in SOCl ₂ (60mL ), reacted at room temperature for 10 min, and then heated to reflux for 2 hours. After completion of the reaction, distillation under reduced pressure to remove SOCl _2, residue was dissolved with EtOAc, washed with H ₂ O, the organic phase was dried over anhydrous Na ₂ SO _4, and concentrated to give an oil which was filtered 9g. The yield was 92%.

Step 3. 2,4-Diamino-6-chloropyrimidine-5-carbonitrile

In a 150 mL round bottom flask, 2,4,6-trichloropyrimidine-5-carbonitrile (9 g, 43.5 mmol), dioxane (40 mL) and aqueous ammonia (36%, 40 mL) were added and heated to 50 ° C. 1 hour. After completion of the reaction, it was cooled to room temperature, and 50 mL of ice water was added thereto, and the mixture was cooled in an ice water bath, and stirred for 1.5 hours. The resulting mixture was then filtered, and the residue was washed with H ₂ O to afford 3.8 g of pale yellow solid. LCMS (ESI): m / z = 170 (M + H) +.

Step 4. (S)-2,4-Diamino-6-(1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethylamine)pyrimidine-5- Nitrile

In a 25 mL round bottom flask, (S)-1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethylamine (150 mg, 0.55 mmol), 2, 4 -Diamino-6-chloropyrimidine-5-carbonitrile (93 mg, 0.55 mmol), KF (64 mg, 1.1 mmol), DIEA (355 mg, 2.75 mmol), DMSO (5 mL), and warmed to 90 ° C for 2 hours. After completion of the reaction, diluted with H ₂ O, EtOAc. The combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered and concentrated. The residue was purified by column chromatography (EtOAc:EtOAc:EtOAc LCMS (ESI): m / z = 406 (M + H) +. _{^{1 H NMR (400MHz, CDCl 3}} ) δ8.71 (s, 1H), 8.02 (s, 1H), 7.79-7.69 (m, 1H), 7.69-7.56 (m, 3H), 7.50-7.38 (m, 1H ), 5.47 (d, J = 6.7 Hz, 1H), 5.31-5.08 (m, 3H), 4.80 (s, 2H), 1.57 (d, J = 7.1 Hz, 3H).

Example 3.

(S)-2,4-diamino-6-(1-(3-chloro-5-(3-fluorophenyl)pyrazole[1,5-a]pyrimidin-6-yl)ethylamine)pyrimidine- 5-carbonitrile

Step 1. Ethyl 3-(3-fluorophenyl)-3-carbonylpropanoate

Potassium tert-butoxide (20 g, 178.6 mmol) was dissolved in 200 mL of THF, cooled to -20 ° C, and ethyl 3-fluorobenzoate (10 g, 59.5 mmol) was added dropwise, and the reaction was maintained for 30 min. Then, ethyl acetate (15.7 g, 178.4 mmol) was slowly added dropwise, and the temperature was maintained for 1 hour after the completion of the dropwise addition. After completion of reaction, 3N aqueous HCl (75mL), remove THF, water phase was extracted with EtOAc distillation under reduced pressure, and the combined organic phase was successively washed with saturated Na ₂ CO ₃ and saturated NaCl solutions, dried over anhydrous Na ₂ SO _4, filtered After concentration, 5 g of an oil was obtained, which was directly used for the next reaction, yield 40%. LCMS (ESI): m / z = 211 (M + H) +.

Step 2. 5-(3-Fluorophenyl)pyrazole [1,5-a]pyrimidine-6-carboxylic acid ethyl ester

In a 250 mL round bottom flask, ethyl 3-(3-fluorophenyl)-3-carbonylpropanoate (5 g, 23.8 mmol), N,N-dimethylformamide dimethyl acetal (2.9 g, 24.4 mmol), 50 mL of toluene, and refluxed for 2 hours. After the reaction was completed, toluene was distilled off under reduced pressure. The obtained residue was dissolved in 100 mL of ethanol, and 3-aminopyrazole (2 g, 24 mmol) and glacial acetic acid (20 mL) were added under stirring, and the mixture was reacted at room temperature for 8 hours. After completion of the reaction, ethanol was evaporated under reduced pressure, and the residue was diluted with H ₂ O, and extracted with CH ₂ Cl ₂ . The obtained organic phase was washed successively with a saturated Na ₂ CO ₃ solution, saturated NaCI solution, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to give a white solid 4.3 g, which was directly used for the next step, yield 63%. LCMS (ESI): m / z = 286 (M + H) +.

Step 3. 3-Chloro-5-(3-fluorophenyl)pyrazole [1,5-a]pyrimidine-6-carboxylic acid ethyl ester

5- (3-fluorophenyl) pyrazolo [1,5-a] pyrimidine-6-carboxylate (4.3g, 15.1mmol) was dissolved in 50 mL CHCl _3, was added NCS (2.1g, 15.7mmol) The temperature was raised to 60 ° C for 6 hours. After completion of the reaction, it was diluted with CH ₂ Cl ₂ and washed successively with 0.1N aqueous HCl and saturated NaCI. The organic phase was dried over anhydrous Na ₂ SO _4, filtered and concentrated to give 4.6 g of a yellow solid, was used directly in the next step, 95% yield. LCMS (ESI): m / z = 320 (M + H) +.

Step 4. 3-Chloro-5-(3-fluorophenyl)pyrazole [1,5-a]pyrimidine-6-carboxylic acid

In a 100 mL round bottom flask, ethyl 3-chloro-5-(3-fluorophenyl)pyrazole [1,5-a]pyrimidine-6-carboxylate (4.6 g, 14.4 mmol), methanol 20 mL, 2N 60 mL of an aqueous NaOH solution was reacted at room temperature for 5 hours. After completion of the reaction, methanol was distilled off under reduced pressure, filtered, and the residue was washed with H ₂ O. The obtained filtrate was adjusted to pH = 3 with 4N HCl solution, and a solid was precipitated, which was filtered to give a solid. LCMS (ESI): m / z = 292 (M + H) +.

Step 5. 3-Chloro-5-(3-fluorophenyl)-N-methoxy-N-methylpyrazole [1,5-a]pyrimidine-6-amide

3-Chloro-5-(3-fluorophenyl)pyrazole [1,5-a]pyrimidine-6-carboxylic acid (2.3 g, 7.9 mmol), DIEA (3 g, 23.3 mmol) dissolved in 20 mL CH ₂ Cl ₂ , N,O-dimethylhydroxylamine hydrochloride (850 mg, 8.7 mmol), HATU (3.3 g, 8.7 mmol) was added under stirring, and the mixture was reacted at room temperature for 2 hours. After completion of the reaction, diluted with CH ₂ Cl _2, the organic phase was washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered and concentrated, residue was purified by column obtained (PE:EtOAc (v / v) = 2:1) A solid of 2 g was obtained with a yield of 76%. LCMS (ESI): m / z = 335 (M + H) +.

Step 6. 1-(3-Chloro-5-(3-fluorophenyl)pyrazole [1,5-a]pyrimidin-6-yl)ethanone

Dissolving 3-chloro-5-(3-fluorophenyl)-N-methoxy-N-methylpyrazole [1,5-a]pyrimidin-6-amide (2 g, 6 mmol) under N2 20 mL of THF was cooled in an ice bath. The solution was slowly added dropwise to a solution of methylmagnesium chloride in THF (3M, 3 mL). The temperature was maintained for 1 hour after the completion of the dropping. After completion of the reaction, the reaction was quenched by carefully adding a saturated NH ₄ Cl solution, and THF was evaporated under reduced pressure. The aqueous phase was extracted with EtOAc, the combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, and concentrated to give a solid 1.5g, 87% yield after filtration. LCMS (ESI): m / z = 290 (M + H) +.

Step 7. (R,E)-N-(1-(3-Chloro-5-(3-fluorophenyl)pyrazole [1,5-a]pyrimidin-6-yl)ethylidene)-2- Methylpropane-2-sulfinamide

(R)-tert-butylsulfinamide (692 mg, 5.7 mmol) was dissolved in 10 mL of THF, and 1-(3-chloro-5-(3-fluorophenyl)pyrazole [1,5-a] was added dropwise. Pyrimidine-6-yl)ethanone (1.5 g, 5.2 mmol) and tetraethyl titanate (2.4 g, 10.5 mmol) in THF (20 mL). Then, the temperature was refluxed for 4 hours. After completion of the reaction, it was cooled to room temperature, a saturated NH ₄ Cl solution was added, and THF was evaporated under reduced pressure. The aqueous phase was extracted with EtOAc, the combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered and concentrated, the resulting residue was purified by column chromatography (PE:EtOAc (v / v) = 1:1) to give a solid 840mg The yield was 41%. LCMS (ESI): m / z = 393 (M + H) +.

Step 8. (R)-N-((S)-1-(3-Chloro-5-(3-fluorophenyl)pyrazole [1,5-a]pyrimidin-6-yl)ethyl)-2 -methylpropane-2-sulfinamide

(R,E)-N-(1-(3-chloro-5-(3-fluorophenyl)pyrazole[1,5-a]pyrimidin-6-yl)ethylene)-protected under nitrogen 2-Methylpropane-2-sulfinamide (840 mg, 2.14 mmol) was dissolved in 10 mL THF and cooled to -40. While maintaining the temperature, a solution of lithium tri-sec-butylborohydride in THF (1 M, 4.3 mL) was slowly added dropwise. The temperature was maintained for 1 hour after the completion of the dropping. After completion of the reaction, saturated NH ₄ Cl solution, stirred for 5min, warmed to room temperature, evaporated under reduced pressure to remove THF, water phase was extracted with EtOAc, the combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered Concentration and purification of the residue (EtOAc: m. LCMS (ESI): m / z = 395 (M + H) +.

Step 9. (S)-1-(3-Chloro-5-(3-fluorophenyl)pyrazole [1,5-a]pyrimidin-6-yl)ethylamine

(R)-N-((S)-1-(3-chloro-5-(3-fluorophenyl)pyrazole [1,5-a]pyrimidin-6-yl)ethyl)-2-methyl The propane-2-sulfinamide (600 mg, 1.52 mmol) was dissolved in 5 mL of MeOH, and concentrated hydrochloric acid (2 mL). After completion of the reaction, it was cooled in an ice bath and adjusted to pH = 9 with a saturated Na ₂ CO ₃ solution. Extracted with EtOAc phase was distilled under reduced pressure to remove methanol, water, and the combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered and concentrated to give a solid 400mg, 91% yield. LCMS (ESI): m / z = 291 (M + H) +.

Step 10. (S)-2,4-Diamino-6-(1-(3-chloro-5-(3-fluorophenyl)pyrazole[1,5-a]pyrimidin-6-yl)ethylamine Pyrimidine-5-carbonitrile

In a 25 mL round bottom flask, (S)-1-(3-chloro-5-(3-fluorophenyl)pyrazole [1,5-a]pyrimidin-6-yl)ethylamine (100 mg, 0.34 mmol) was added. , 2,4-diamino-5-cyano-6-chloropyrimidine (58 mg, 0.34 mmol), KF (40 mg, 0.69 mmol), DIEA (219 mg, 1.7 mmol), DMSO (5 mL), warmed to 90 ° C Reaction for 2 hours. After completion of the reaction, diluted with H ₂ O, EtOAc. The combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered and concentrated. The residue was purified by column chromatography (EtOAc:EtOAc:EtOAc LCMS (ESI): m / z = 406 (M + H) +. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.70 (s, 1H), 8.03 (s, 1H), 7.72-7.52 (m, 2H), 7.45-7.28 (m, 2H), 5.44-5.29 (m, 1H) ), 5.26-5.13 (m, 1H), 5.09 (s, 2H), 4.78 (s, 2H), 1.55 (d, J = 7.0 Hz, 3H).

Example 4.

(S)-4-amino-6-(1-(3-chloro-5-(3-fluorophenyl)pyrazole[1,5-a]pyrimidin-6-yl)ethylamine)pyrimidine-5- Nitrile

In a 25 mL round bottom flask, (S)-1-(3-chloro-5-(3-fluorophenyl)pyrazole [1,5-a]pyrimidin-6-yl)ethylamine (100 mg, 0.34 mmol) was added. , 4-amino-5-cyano-6-chloropyrimidine (53 mg, 0.34 mmol), KF (40 mg, 0.69 mmol), DIEA (219 mg, 1.7 mmol), DMSO (5 mL), warmed to 90 ° C for 2 hours . After completion of the reaction, diluted with H ₂ O, EtOAc. The combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered and concentrated. The residue was purified by column chromatography (EtOAc:EtOAc:EtOAc) LCMS (ESI): m / z = 409 (M + H) +. _{^{1 H NMR (400MHz, CDCl 3}} ) δ8.67 (s, 1H), 8.10-7.98 (m, 2H), 7.68-7.54 (m, 2H), 7.42-7.28 (m, 2H), 5.54 (d, J =6.0 Hz, 1H), 5.41 (s, 2H), 5.22-5.10 (m, 1H), 1.57 (d, J = 7.1 Hz, 3H).

Example 5.

(S)-2-Amino-4-((1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethyl)amino)-6-methylpyrimidine- 5-carbonitrile

Step 1. 4-Chloro-5-iodo-6-methylpyrimidin-2-amine

4-Chloro-6-methylpyrimidin-2-amine (3 g, 21.0 mmol) was dissolved in 40 mL of glacial acetic acid in a 100 mL round bottom flask and cooled to below 10 ° C in an ice bath. NIS (2.4 g, 10.6 mmol) was added to the solution. After 1 hour, NIS (2.4 g, 10.6 mmol) was added. After 1 hour, the reaction was allowed to naturally rise to room temperature for 8 hours. After the reaction was completed, the reaction solution was poured into ice water and extracted with EtOAc. The organic phase was washed with 5% Na ₂ SO ₃ solution, 10% NaHCO ₃ solution and saturated NaCl solution, dried over anhydrous Na ₂ SO ₄ solvent was removed under reduced pressure and concentrated to give 4.9g solid, i.e. the title compound, yield 87%, LCMS (ESI): m / z = 270 (m + H) +.

Step 2. 2-Amino-4-chloro-6-methylpyrimidine-5-carbonitrile

4-Chloro-5-iodo-6-methylpyrimidin-2-amine (4.9 g, 18.2 mmol), Zn(CN) ₂ (4.3 g, 36.6 mmol) in a 150 mL round bottom flask under nitrogen. , Pd(PPh ₃ ) ₄ (2.1 g, 1.8 mmol), CuI (1.7 g, 8.9 mmol) and DMF (60 mL), heated to 80 ° C for 16 hours. After the reaction was completed, diluted with EtOAc EtOAc. The resulting filtrate was added H ₂ O, extracted with EtOAc, the organic phase was washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered and concentrated, residue was purified by column obtained (PE:EtOAc (v / v) = 5:1) A solid of 1.1 g was obtained with a yield of 36%. LCMS (ESI): m / z = 169 (M + H) +.

Step 3. (S)-2-Amino-4-((1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethyl)amino)-6-A Pyrimidine-5-carbonitrile

In a 25 mL round bottom flask, (S)-1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethylamine (100 mg, 0.37 mmol), 2-amino 4-chloro-6-methylpyrimidine-5-carbonitrile (62 mg, 0.37 mmol), KF (43 mg, 0.74 mmol), DIEA (232 mg, 1.8 mmol), DMSO (5 mL), warmed to 90 ° C for 2 hours . After completion of the reaction, diluted with H ₂ O, EtOAc. The combined organic phases were washed with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, filtered and concentrated. The residue was purified by column chromatography (EtOAc:EtOAc) LCMS (ESI): m / z = 405 (M + H) +. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.70 (s, 1H), 8.03 (s, 1H), 7.79-7.31 (m, 5H), 5.76 (d, J = 6.7 Hz, 1H), 5.66-4.71 ( m, 3H), 2.39 (s, 3H), 1.63 (d, J = 7.1 Hz, 3H).

The compounds of the invention were used to modulate kinase activity and inhibit cell proliferation by experiments as described below.

Example A: Determination of PI3K kinase activity

The kinase activity of PI3K was tested by the ADP-Glo method. ADP-Glo is from Promega (#V9101). P110/p85 was from Millipore (#14-604-K). Compound IC ₅₀ of each compound was assessed by determining the concentration of point 10, which is a starting concentration of 1M, and then 3-fold serial dilutions. The test buffer was 50 mM HEPES pH 7.5, 3 mM MgCl2, 1 mM EGTA, 100 mM NaCl, 0.03% CHAPS, 2 mM DTT. The final kinase concentration was PI3K 17 nM with a final PIP2 concentration of 50 M and a final ATP concentration of 25M.

1. Preparation of 10 L of kinase reaction solution: To a reaction well of a 384-well plate (Corning #4512), 2.5 l of a test compound solution, 2.5 l of a kinase solution, and 5 l of a substrate solution were added.

2. Cover the 384-well plate and incubate for 60 minutes at room temperature.

3. Transfer 5 l of reaction solution from each well to a new 384-well plate well.

4. Add 5 l of ADP-Glo reagent to the 384-well plate reaction well to stop the reaction.

5. Gently shake for 40 minutes on the shaker.

6. Add 10l of kinase assay reagent to each well, shake for 1 minute, and let stand for 30 minutes at room temperature.

8. Read the sample luminescence value with Synegy.

9. A data processing Excel, for curve fitting using XLFit software obtained IC ₅₀ values for each compound (Table 1).

Example B. Detection of Cellular Inhibitory Activity of Compounds

Compounds were tested for inhibition of proliferation of lymphoma cell line SU-DHL-6 (ATCC, Cat. No.: CRL-2959) by the CellTiter-Glo method. The cell culture fluid used in the experiment was RPMI 1640 (Invitrogen, catalog number: 11875-093). The experiment used 10% fetal bovine serum (Invitrogen, catalog number: 10099-141).

One hundred microliters of the culture solution containing 15,000 cells was dispensed into wells of a 96-well culture plate (Corning #3903), and placed in a carbon dioxide incubator for overnight culture. On the next day, 0.5 μl of test compound (8 consecutive concentration gradients in DMSO) was added to each well, and two replicates were set for each concentration, and cell-free wells (blank control) and DMSO wells (solvent control) were set. . After the administration, the cells were further cultured for 72 hours under conditions of 37 ° C and 5% carbon dioxide. Finally, 100 μl of CellTiter-Glo reagent (Promega, catalog number: G7571) was added to each well, and the luminescence signal was detected with Flex Station 3 (Molecular Devices), and the IC ₅₀ value of the compound for inhibition of cell proliferation was calculated using XLfit software (Table 1).

Table 1. Kinase activity and cell viability of compounds

A: <20nM; B: 20-100nM; C:>100-1000nM

As can be seen from the results in Table 1, the compounds of the present invention have a good kinase inhibitory activity against PI3K. The compounds of the invention are also effective in inhibiting the growth of lymphoma cells.

Claims (10)

1. A compound of formula (I):

   

   Or a pharmaceutically acceptable salt thereof, wherein:

   R ^{1 is} selected from H, halogen, cyano, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl, wherein said alkyl, alkenyl, alkynyl, An aryl, heteroaryl, cycloalkyl, or heterocycloalkyl group is optionally substituted with from 1 to 5 R ^1a ;

   R ^{1a is} selected from the group consisting of H, anthracene, halogen, cyano, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl, wherein said alkyl, alkenyl, alkyne Alkyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl optionally substituted by halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , alkyl, alkenyl, alkynyl, cycloalkyl , or a heterocycloalkyl group;

   R ^{2 is} selected from aryl, heteroaryl, cycloalkyl, or heterocycloalkyl, wherein R ^{2 is} optionally substituted with from 1 to 5 R ^2a ;

   R ^{2a is} selected from the group consisting of H, halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , NR ^b C(O)R ^d , NR ^b C(O)OR ^d , NR ^b C(O)NR ^b R ^c , NR ^b S(O) ₂ R ^d , NR ^b S(O) ₂ NR ^b R ^c , C(O)NR ^b R ^c , C(O)OR ^d , S(O) ₂ NR ^b R ^c , C (O) R ^d , S(O) ₂ R ^d , P(O)R ^b R ^c , alkyl, alkenyl, or alkynyl;

   R ^{3 is} selected from the group consisting of H, anthracene, alkyl, alkenyl, alkynyl, haloalkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cyanoalkyl, cycloalkyl, or cycloalkylalkyl ;

   R ^{4 is} selected from the group consisting of H, alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, cyanoalkyl, cycloalkyl, or cycloalkylalkyl;

   R ^{5 is} selected from H, halogen, cyano, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl, wherein said alkyl, alkenyl, alkynyl, An aryl group, a heteroaryl group, a cycloalkyl group, or a heterocycloalkyl group may be substituted with from 1 to 3 R ^5a ;

   R ^{5a is} selected from the group consisting of H, hydrazine, halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , NR ^b C(O)R ^d , NR ^b C(O)OR ^d , NR ^b C(O)NR ^b R ^c , NR ^b S(O) ₂ R ^d , NR ^b S(O) ₂ NR ^b R ^c , C(O)NR ^b R ^c , C(O)OR ^d , S(O) ₂ NR ^b R ^c , C(O)R ^d , S(O) ₂ R ^d , P(O)R ^b R ^c , alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl ;

   R ⁶ and R ^{7 are} each selected from H, an amino group, or an alkyl group;

   R ^a , R ^b , R ^c , and R ^{d are} independently selected from the group consisting of H, alkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cyanoalkyl, alkenyl, alkynyl, cycloalkyl, heterocycle An alkyl group, an aryl group, or a heteroaryl group;

   Or R ^b and R ^c together with the nitrogen atom to which they are attached form a 4 to 7 membered heterocycloalkyl group, optionally substituted with 1 to 3 R ^e ;

   R ^{e is} selected from the group consisting of H, halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , NR ^b C(O)R ^d , NR ^b C(O)OR ^d , NR ^b C(O)NR ^b R ^c , NR ^b S(O) ₂ R ^d , NR ^b S(O) ₂ NR ^b R ^c , C(O)NR ^b R ^c , C(O)OR ^d , S(O) ₂ NR ^b R ^c , C (O) R ^d , S(O) ₂ R ^d , P(O)R ^b R ^c , alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl.

   Wherein any of the hydrogen atoms in the compound of formula (I) may be deuterium.
2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which has the structure of formula (II),

   

   among them:

   R ^{1 is} selected from H, halogen, cyano, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl, wherein said alkyl, alkenyl, alkynyl, An aryl, heteroaryl, cycloalkyl, or heterocycloalkyl group is optionally substituted with from 1 to 5 R ^1a ;

   R ^{1a is} selected from the group consisting of H, anthracene, halogen, cyano, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl, wherein said alkyl, alkenyl, alkyne Alkyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl optionally substituted by halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , alkyl, alkenyl, alkynyl, cycloalkyl , or a heterocycloalkyl group;

   R ^{2a is} selected from the group consisting of H, halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , NR ^b C(O)R ^d , NR ^b C(O)OR ^d , NR ^b C(O)NR ^b R ^c , NR ^b S(O) ₂ R ^d , NR ^b S(O) ₂ NR ^b R ^c , C(O)NR ^b R ^c , C(O)OR ^d , S(O) ₂ NR ^b R ^c , C (O) R ^d , S(O) ₂ R ^d , P(O)R ^b R ^c , alkyl, alkenyl, or alkynyl;

   R ^{3 is} selected from the group consisting of H, anthracene, alkyl, alkenyl, alkynyl, haloalkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cyanoalkyl, cycloalkyl, or cycloalkylalkyl ;

   R ⁶ and R ^{7 are} each selected from H, an amino group, or an alkyl group;

   R ^a , R ^b , R ^c , and R ^{d are} independently selected from the group consisting of H, alkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, cyanoalkyl, alkenyl, alkynyl, cycloalkyl, heterocycle An alkyl group, an aryl group, or a heteroaryl group;

   Or R ^b and R ^c together with the nitrogen atom to which they are attached form a 4 to 7 membered heterocycloalkyl group, optionally substituted with 1 to 3 R ^e ;

   R ^{e is} selected from the group consisting of H, halogen, cyano, OR ^a , SR ^a , NR ^b R ^c , NR ^b C(O)R ^d , NR ^b C(O)OR ^d , NR ^b C(O)NR ^b R ^c , NR ^b S(O) ₂ R ^d , NR ^b S(O) ₂ NR ^b R ^c , C(O)NR ^b R ^c , C(O)OR ^d , S(O) ₂ NR ^b R ^c , C (O) R ^d , S(O) ₂ R ^d , P(O)R ^b R ^c , alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;

   n is 1, 2, or 3.
3. A compound according to any one of claims 1 to 2, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

   (S)-4-Amino-6-(1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethylamine)pyrimidine-5-carbonitrile

   (S)-2,4-diamino-6-(1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethylamine)pyrimidine-5-carbonitrile

   (S)-2,4-diamino-6-(1-(3-chloro-5-(3-fluorophenyl)pyrazole[1,5-a]pyrimidin-6-yl)ethylamine)pyrimidine- 5-carbonitrile

   (S)-4-amino-6-(1-(3-chloro-5-(3-fluorophenyl)pyrazole[1,5-a]pyrimidin-6-yl)ethylamine)pyrimidine-5- Nitrile

   (S)-2-Amino-4-((1-(3-chloro-5-phenylpyrazole[1,5-a]pyrimidin-6-yl)ethyl)amino)-6-methylpyrimidine- 5-carbonitrile
4. A pharmaceutical composition comprising a compound of any one of claims 1-3, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient.
5. Use of a compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 4, for the preparation of a medicament for the treatment of an effective dose of a disease associated with PI3K.
6. The use according to claim 5, wherein the PI3K-related disease is selected from the group consisting of cancer, Inflammation, and autoimmune diseases.
7. The use according to claim 6, wherein said cancer is selected from the group consisting of acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, Cellular lymphoma, follicular lymphoma, B-cell lymphoma, T-cell lymphoma, and diffuse large B-cell lymphoma.
8. The use according to claim 6, wherein said autoimmune disease and inflammatory disease are selected from the group consisting of skin inflammation, rheumatoid arthritis, allergic rhinitis, asthma, Kline disease, chronic obstructive pulmonary disease, systemic Lupus, psoriasis, multiple sclerosis, activated PI3K syndrome, and Sjogren syndrome.
9. Use of a compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 4 in combination with one or more drugs for the treatment of cancer, inflammation and autoimmunity Sexual disease. The combination drug described therein includes a small molecule compound drug and a macromolecular antibody drug. Small molecule compounds are selected from various kinase inhibitors such as BTK inhibitors and other non-kinase small molecule inhibitors; and macromolecular drugs include anti-CD20, anti-CTLA4, anti-PD-1, anti-PD-L1 antibodies and the like.
10. A method of preparing a compound of any of claims 1-3 consisting of the following steps:

    1). Starting material I-1 and N,N-dimethylformamide dimethyl acetal are refluxed in toluene to give I-2.

    

    2). Compound I-2 is reacted with 3-aminopyrazole in acetic acid to obtain the pyrimidine pyrimidine I-3.

    

    3). Chlorination of I-3 with NCS gives chlorinated product I-4.

    

    4) The ester group in .I-4 is hydrolyzed with a base to form a carboxylic acid I-5.

    

    5). I-5 is condensed with N,O-dimethylhydroxyammonia to give amide compound I-6.

    

    6) Formatting reagent R ³ MgCl is added to the amide bond in I-6 to form ketone I-7.

    

    7). Condensation of ketone I-7 with (R)-sulfinamide affords I-8.

    

    8) The imino group in .I-8 is reduced with lithium tris(hydroxy)borohydride to give the sulfinamide compound I-9 in the S configuration.

    

    9) The sulfinyl group in I-9 was removed with hydrochloric acid to give the free amino group I-10.

    

    10). I-10 is reacted with YL (Y is a R ⁵ , R ⁶ , and R ⁷ substituted pyrimidine, and L is a leaving group) to give the final product (I).