WO2016119707A1

WO2016119707A1 - Novel heteroaryl and heterocycle compounds, compositions and methods

Info

Publication number: WO2016119707A1
Application number: PCT/CN2016/072474
Authority: WO
Inventors: Wei-Guo Su; Guangxiu Dai; Hong Jia; Zhulin ZHANG; Jianyang WENG; Jennifer Diane Venable; Scott Damian BEMBENEK; Wenying Chai; Steven Paul MEDUNA; John Matthew KEITH; Wendy Eccles; Alec Donald Lebsack; William Moore Jones; Russell Christopher SMITH
Original assignee: Hutchison Medipharma Ltd
Current assignee: Hutchmed Ltd
Priority date: 2015-01-29
Filing date: 2016-01-28
Publication date: 2016-08-04
Anticipated expiration: 2017-07-29
Also published as: TW201639845A; AR103566A1

Abstract

The invention relates to novel heteroaryl and heterocycle compounds of formula I and pharmaceutical compositions comprising them, uses and methods thereof for inhibiting the activity of PI₃K and for treating inflammatory and autoimmune diseases and cancer.

Description

NOVEL HETEROARYL AND HETEROCYCLE COMPOUNDS, COMPOSITIONS AND METHODS

FIELD OF THE INVENTION

This invention relates generally to the field of medicine and, more specifically, to novel heteroaryl and heterocycle compounds and pharmaceutical compositions comprising them, uses and methods thereof for inhibiting the activity of PI₃K and for treating inflammatory and autoimmune diseases and cancer.

BACKGROUND OF THE INVENTION

Phosphoinositide 3-kinases (PI 3-kinases or PI₃Ks) are a family of enzymes involved in cellular functions such as cell growth, proliferation, differentiation, motility, survival and intracellular trafficking. After exposure of cells to various biological stimuli, PI₃Ks primarily phosphorylate phosphatidylinositol-4, 5-bisphosphate (PtdIns (4, 5) P2, PIP2) at the 3′-OH position of the inositol ring to generate phosphatidylinositol-3, 4, 5-trisphosphate (PtdIns (3, 4, 5) P3, PIP3) which has an important role as second messengers by working as a docking platform for lipid-binding domains, such as the pleckstrin homology (PH) domains of various cellular proteins. These include kinases (such as 3-phosphoinositide-dependent protein kinase 1 (PDK1) and protein kinase B (PKB) /Akt) that trigger downstream kinase cascades, and guanine-nucleotide exchange factors (such as Vav and P-Rex) that control the activity of small GTPases (T Rückle, M. K. et al. Nature Reviews Drug Discovery, 2006, 5, 903-918) .

Based on sequence homology and lipid substrate specificity, the PI₃K family is divided into three classes: I, II, and III. The most studied and the focus of this invention, the class I PI₃Ks, are heterodimeric proteins, each containing a smaller regulatory domain and a larger 110 kDa catalytic domain which occur in four isoforms differentiated as p110α, p110β, p110γ and p110δ (T.J. Sundstrom. et al Org. Biomol. Chem., 2009, 7, 840–850) . Among them, p110α, p110β and p110δ together, termed as the class IA PI₃K, bind to p85 regulatory subunit and are primarily activated by protein tyrosine kinase-coupled receptors (RTK) and/or Ras proteins, whereas PI₃Kγ as the sole class IB member, binds to one of two noncatalytic subunits, p101 or p87, is activated by G-protein coupled receptors (GPCRs) through direct interaction with G-protein β γ dimers and Ras proteins, which are widely implicated in various aspects of immune function and regulation.

All four class I catalytic PI₃K isoforms show a characteristic expression pattern in vivo. p110α and p110β are ubiquitously expressed, while p110 γ and p110δ are found predominantly in leukocytes, endothelial cells and smooth muscle cells (T.J. Sundstrom. et al Org. Biomol. Chem., 2009, 7, 840–850) . Deletion of the class IA isoform p110α or β induces embryonic lethality (E9.5-E10) (Bi L, Okabe I. et al . J Biol Chem, 1999, 274: 10963–8； Bi L, Okabe I. et al. Mamm Genome. 2002, 13, 169–72) . p110γ-deficient mice develop and reproduce normally, although they have suboptimal immune responses because of defects in T-cell activation as well as in neutrophil and macrophage migration. The loss of p110δ mice are also viable and fertile but exhibit significant defects in T, B cell activation (AGhigo. et al. BioEssays 2010, 32: 185–196) .

Dysregulation and overactivation of the PI₃K/AKT pathway has been firmly established in cancer cells. In principle, modulating PI₃K and thus controlling PIP3 levels should regulate AKT activity and ultimately suppress tumor growth. The expression of PI₃Kδ is generally restricted to hematopoietic cell types. The p110δ isoform is constitutively activated in B cell tumors. Genetic and pharmacologic approaches that specifically inactivate the p110δ isoform have demonstrated its important role for the treatment of B cell malignancy (B.J. Lannutti. et al. Blood. 2011, 117, 591-594) . Previous studies have shown that CAL-101, a potent and selective p110 inhibitor, has broad antitumor activity against cancer cells of hematologic origin. (Lannutti B.J. Am Soc Hematol. 2008； 112. Abstract 16； Flinn I. W. et al. J. Clin. Oncol. 2009； 27 (A3543) )

In addition to cancer, PI₃K has also been suggested as a target for inflammatory and autoimmune disorders. The isoforms p110δ and p110γ are mainly expressed in cells of the immune system and contributes to innate and adaptive immunity. p110δ and p110γregulate diverse immune cell function. For example, inhibition of p110δ leads to suppression of B-cell activation and function, suppression of T-lymphocyte proliferation, T- cell trafficking, and Th1-Th2 differentiation and Treg function. Inhibition of both p110δ and p110γ results in inhibition of neutrophil (leukocyte) chemotaxis, inhibition of mast cell activation, intact macrophage phagocytosis and endothelium activation. Inhibition of p110γ could activate microglial (C. Rommel. et al. Current Topics in Microbiology and Immunology, 2010, 1, 346, 279-299) . So isoform-specific p110δ or p110γ inhibitors are expected to have therapeutic effects on these diseases without interfering with general PI₃K signaling critical to the normal function of other cellular systems. p110δ and p110γ supporting the hypothesis that p110γ alone, p110δ alone, or dual-blockade of both, all present a unique therapeutic opportunity in that pharmacological inhibition, but the two PI₃K isoforms simultaneously may yield more superior clinical results in the treatment of a variety of complex immune-mediated inflammatory diseases. In the case of RA, Phosphoinositide 3-kinases (PI₃Ks) , most notably PI₃Kδ and PI₃Kγ, have crucial and specific roles at all stages of disease progression: in antigen signalling in B and T cells, and in signalling downstream of FcRs, cytokine receptors and chemokine receptors in mast cells, macrophages, neutrophils and synoviocytes (C. Rommel. et al. Nature Reviews Immunology, 2007, 7, 191-201) . Although the pathogenesis of RA is not yet completely understood, chemokines and other chemoattractants have been detected in the inflamed joint and are responsible for the recruitment of leukocytes into the joints. Amongst these, neutrophils constitute the most abundant population and are capable of inducing inflammatory response and tissue damage (T Rückle, M.K. et al. Nature Reviews Drug Discovery, 2006, 5, 903-918) . Blockade of hematopoietic PI₃Kγ and/or PI₃Kδ can potently suppresses neutrophil chemotaxis and, in turn, the progression of joint inflammation and cartilage erosion.

Novel compounds are disclosed which in some instances are inhibitors of PI₃Ks kinase activity including p110δ, p110γ, p110α, and p110β. These compounds therefore have potential therapeutic benefit in the treatment of a variety of diseases associated with inappropriate p110δ, p110γ, p110α, and p110β activity, such as cancer, inflammatory, allergic and autoimmune diseases and leukemia etc, in particular systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) , allergic disorders, respiratory diseases like asthma and chronic obstructive pulmonary disease (COPD) , multiple sclerosis, all pathologic conditions whose onset and/or progression is driven by an inflammatory insult, such as myocardial infarction and cancer. Examples of inhibitors of PI₃Ks have been described in, for example, published application Nos. WO 2014/015523 A1, WO 2014/015675 A1, and WO 2014/015830 A1, the content of each of these publications is incorporated by reference herein in its entirety.

SUMMARY OF THE I NVENTION

The present invention provides a compound of formula I:

and/or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein Z₁, Z₂, Z₃, Z₄, W, m, R₁, R₂, R₃, R₄, and R₅ are as defined in the detailed description of the invention.

Also provided is a pharmaceutical composition, comprising at least one compound of formula I (e.g., any of the compounds described herein) and/or at least one pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient (e.g., a pharmaceutically acceptable carrier) .

Also provided is a method of in vivo or in vitro inhibiting the activity of PI₃K, comprising contacting the PI₃K with an effective amount of at least one compound of formula I (e.g., any of the compounds described herein) and/or at least one pharmaceutically acceptable salt thereof.

Also provided is a method of treating a disease responsive to inhibition of PI₃K in a subject, comprising administering to the subject in need thereof a therapeutically effective amount of at least one compound of formula I (e.g., any of the compounds described herein) and/or at least one pharmaceutically acceptable salt thereof.

Also provided is at least one compound of formula I (e.g., any of the compounds described herein) and/or at least one pharmaceutically acceptable salt thereof described herein for treating a disease responsive to inhibition of PI₃K.

Also provided is a use of at least one compound of formula I (e.g., any of the compounds described herein) and/or at least one pharmaceutically acceptable salt thereof described herein in the manufacture of a medicament for treating a disease responsive to inhibition of PI₃K.

The subject described herein can be human.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

As used in the present specification, the following words, phrases and symbols are generally intended to have the meanings as set forth below, except to the extent that the context in which they are used indicates otherwise. The following abbreviations and terms have the indicated meanings throughout:

A dash ( “- “) that is not between two letters or symbols is used to indicate a point of attachment for a substituent. For example, -CONH₂ is attached through the carbon atom. However, when the point of attachment of a group is apparent to those skilled in the art, e.g., a halo substituent, the "-" sign may be omitted.

The term "alkyl" herein refers to a straight or branched saturated hydrocarbon radical, containing 1-18, preferably 1-12, more preferably 1-6, especially 1-4 carbon atoms. For example, “C_1-6 alkyl” refers to an alkyl containing 1-6 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl ( "Me" ) , ethyl (" Et" ) , n-propyl ( "n-Pr" ) , i-propyl ( "i-Pr" ) , n-butyl ( "n-Bu" ) , i-butyl ( "i-Bu" ) , s-butyl ( "s-Bu" ) and t-butyl ( "t-Bu" ) .

The term “alkenyl” as used herein refers to a straight or branched hydrocarbon radical, containing one or more, for example 1, 2 or 3, C＝C double bonds, and 2-10, preferably 2-6, more preferably 2-4 carbon atoms. For example, “C_2-6 alkenyl” refers to an alkenyl containing 2-6 carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, and 2-butenyl.

The term “alkynyl” as used herein refers to a straight or branched hydrocarbon radical, containing one or more, for example 1, 2 or 3, C≡C triple bonds, and 2-10, preferably 2-6, more preferably 2-4 carbon atoms. For example, “C_2-6 alkynyl” refers to an alkyl containing 2-6 carbon atoms. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, and 2-butynyl.

The term “halo” as used herein includes fluoro, chloro, bromo, and iodo, and the term “halogen” as used herein includes fluorine, chlorine, bromine, and iodine.

The term “haloalkyl” as used herein refers to an alkyl radical, as defined herein, in which one or more, for example 1, 2, 3, 4, or, 5, hydrogen atoms are replaced with halogen atom, and when two or more hydrogen atoms are replaced with halogen atom, the halogen atoms are the same as one another or different from one another. In certain embodiments, the term “haloalkyl” as used herein refers to an alkyl radical, as defined herein, in which two or more, such as 2, 3, 4, or 5 hydrogen atoms are replaced with halogen atoms, the halogen atoms are all the same as one another. In other embodiments, the term “haloalkyl” as used herein refers to an alkyl radical, as defined herein, in which two or more, such as 2, 3, 4, or 5 hydrogen atoms are replaced with halogen atoms, the halogen atoms are not all the same as one another. Examples of haloalkyl groups include, but are not limited to, -CF₃, -CHF₂, -CH₂CF₃, and the like.

The term “alkoxy” as used herein refers to the group –O-alkyl, wherein the alkyl is as defined above. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propyloxy, i-propyloxy, n-butyloxy, i-butyloxy, t-butyloxy, pentyloxy, hexyloxy, including their isomers.

The term “cycloalkyl” as used herein refers to saturated and partially unsaturated cyclic hydrocarbon radical which may have one or more, preferably 1 or 2 rings, having 3 to 12, preferably 3 to 8, more preferably 3 to 6 ring carbon atoms. For example, “C_3-8 cycloalkyl” refers to a cycloalkyl containing 3-8 carbon atoms in the ring. The ring of the cycloalkyl group may be saturated or has one or more, for example, one or two double bonds (i.e. partially unsaturated) , but not fully conjugated, and not aryl as defined herein. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

The term “aryl” as used herein refers to carbocyclic hydrocarbon radical of a monocyclic ring or fused rings containing 6-14 ring carbon atoms, preferably 6-12 ring carbon atoms, wherein at least one ring is aromatic. For example, the aryl includs phenyl or fused bicyclic carbocyclic hydrocarbon radical containing 8-12 ring carbon atoms, wherein at least one ring is aromatic. Examples of aryl groups include, but are not limited to, phenyl, naphthalenyl, 1, 2, 3, 4-tetrahydronaphthalenyl, indenyl, indanyl, azulenyl.

The term “heteroaryl” as used herein refers to:

monocyclic aromatic hydrocarbon radical having 5, 6 or 7 ring atoms, preferably having 6 ring atoms, and containing one or more, for example 1, 2 or 3, preferably 1 or 2 heteroatoms independently selected from N, O, and S (preferably N) in the ring, with the remaining ring atoms being carbon； and

bicyclic aromatic hydrocarbon radical having 8-12 ring atoms, preferably having 9 or 10 ring atoms, and containing one or more, for example, 1, 2, 3 or 4, preferably 1 or 2 heteroatoms independently selected from N, O, and S (preferably N) in the rings, with the remaining ring atoms being carbon, wherein at least one of the rings is aromatic. For example, the bicyclic heteroaryl includes a 5-to 6-membered heterocyclic aromatic ring fused to a 5-to 6-membered cycloalkyl ring.

When the total number of S and O atoms in the heteroaryl group exceeds 1, those heteroatoms are not adjacent to one another. In some embodiments, the total number of S and O atoms in the heteroaryl group is not more than 2. In some embodiments, the total number of S and O atoms in the heteroaryl group is not more than 1.

The heteroaryl group also includes those wherein the N heteroatom occurs as N-oxide, such as pyridyl N-oxides.

Examples of the heteroaryl group include, but are not limited to, pyridyl, pyridyl N-oxide, such as pyrid-2-yl, pyrid-3-yl, pyrid-4-yl or N-oxide thereof； pyrazinyl, such as pyrazin-2-yl, pyrazin-3-yl； pyrimidinyl, such as pyrimidin-2-yl, pyrimidin-4-yl； pyrazolyl, such as pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, pyrazol-5-yl； imidazolyl, imidazol-2-yl, imidazolin-4-yl； oxazolyl； isoxazolyl； thiazolyl； isothiazolyl； thiadiazolyl； tetrazolyl, such as tetrazol-5-yl； triazolyl； thienyl； furyl； pyranyl； pyrrolyl； pyridazinyl； bezodioxolyl, such as benzo [d] [1, 3] dioxolyl； benzoxazolyl, such as benzo [d] oxazolyl； imidazopyridinyl, such as imidazo [1, 2-a] pyridinyl； triazolopyridinyl, such as [1, 2, 4] triazolo [4, 3-a] pyridinyl and [1, 2, 4] triazolo [1, 5-a] pyridinyl； indazolyl； pyrrolopyrimidinyl, such as pyrrolo [1, 5-a] pyrimidinyl； tetrazolopyridinyl, such as tetrazolo [1, 5-a] pyridinyl； pyrazolopyrimidinyl, such as pyrazolo [1, 5-a] pyrimidinyl； benzothienyl； benzofuryl； benzoimidazolinyl； indolyl； indolinyl； purinyl, such as 9H-purinyl and 7H-purinyl； quinolinyl, isoquinolinyl, 1, 2, 3, 4-tetrahydroquinolinyl and 5, 6, 7, 8-tetrahydroisoquinolinyl.

The term “heterocycle” as used herein refers monocyclic, bicyclic or tricyclic saturated or partially unsaturated cyclic hydrocarbon radical having 3-14 ring atoms, preferably having 4-12 ring atoms, and containing one or more, for example 1, 2, 3 or 4, preferably 1 or 2 heteroatoms independently selected from N, O, and S in the rings, with the remaining ring atoms being carbon. The rings of the heterocycle group may be saturated or has one or more, for example, one or two double bonds (i.e. partially unsaturated) , but not fully conjugated. The point of the attachment may be carbon or heteroatom in the heterocycle group. However, any of the rings in the heterocycle group is not aromatic so that the heterocycle group is not a heteroaryl as defined herein.

In some embodiments, “heterocycle” refers to 4-to 8-membered, especially 4-, 5-or 6-membered monocyclic heterocycle group containing 1 or 2 heteroatoms independently selected from N, O, and S in the ring, with the remaining ring atoms being carbon.

The heterocycle group also includes those wherein the N or S heteroatom occurs as oxide thereof.

Examples of heterocycle include, but are not limited to, oxetanyl, such as oxetan-2-yl or oxetan-3-yl； azetidinyl, such as azetidin-2-yl or azetidin-3-yl； pyrrolidinyl, such as pyrrolidin-1-yl, pyrrolidin-2-yl, pyrrolidin-3-yl； tetrahydrofuranyl, such as tetrahydrofuran-2-yl, tetrahydrofuran-3-yl； tetrahydropyranyl, such as tetrahydropyran-2-yl, tetrahydropyran-3-yl, tetrahydropyran-4-yl； dioxolanyl, such as 1, 3-dioxolanyl； dioxanyl, sush as 1, 4-dioxanyl, 1,3-dioxanyl； morpholinyl, morpholinyl-N-oxide, such as morpholin-2-yl, morpholin-3-yl, morpholin-4-yl (morpholino) (numbered wherein the oxygen is assigned priority 1) ； thiomorpholinyl, 1-oxo-thiomorpholin-4-yl, 1, 1-dioxo-thiomorpholin-4-yl； imidazolinyl, such as imidazolidin-2-yl, imidazolidin-4-yl； pyrazolidinyl, such as pyrazolidin-2-yl, pyrazolidin-3-yl； piperidinyl or piperidinyl N-oxide, such as piperidin-1-yl and piperidin-2-yl, piperidin-3-yl, piperidin-4-yl or N-oxide thereof； and piperazinyl, such as piperazin-1-yl, piperazin-2-yl, piperazin-3-yl； octahydropyrrolo [3, 4-b] pyrrolyl.

The term “oxo” as used herein refers to the ＝O radical.

The term “optional” or “optionally” means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances in which it does not. For example, “alkyl optionally substituted with -OH” encompasses both “unsubstituted alkyl” and “alkyl substituted with -OH” as defined herein. It is understood by those skilled in the art, with respect to any group containing one or more substituents, that such groups are not intended to introduce any substitution or substitution patterns that are sterically impractical, chemically incorrect, synthetically non-feasible and/or inherently unstable.

The term “substituted” or “substituted with……” , as used herein, means that one or more hydrogens on the designated atom or group are replaced with one or more selections from the indicated group of substituents, provided that the designated atom's normal valence is not exceeded. When a substituent is oxo (i.e., ＝O) , then 2 hydrogens on a single atom are replaced. Combinations of substituents and/or variables are permissible only if such combinations result in a chemically correct and stable compound. A chemically correct and stable compound is meant to imply a compound that is sufficiently robust to survive isolation from a reaction mixture, and subsequent formulation as an agent having at least practical utility. Unless otherwise specified, substituents are named into the core structure. For example, it is to be understood that when (cycloalkyl) alkyl is listed as a possible substituent, the point of attachment of this substituent to the core structure is in the alkyl portion.

The term “substituted with one or more substitutents” as used herein means that one or more hydrogens on the designated atom or group are independently replaced with one or more selections from the indicated group of substituents. In some embodiments, “substituted with one or more substitutents” means that the designated atom or group is substituted with 1, 2, 3, or 4 substitutents independently selected from the indicated group of substituents.

Some of the compounds of the present invention can exist as stereoisomers including optical isomers. The invention includes all such stereoisomers, as pure individual stereoisomer preparations and as enriched preparations of each, and as the racemic mixtures of such stereoisomers as well as the individual enantiomers and diastereomers that may be separated according to methods that are well-known to those of skill in the art.

It will be appreciated by those skilled in the art that some of the compounds of formula I may contain one or more chiral centers and therefore exist in two or more stereoisomeric forms. The racemates of these isomers, the individual isomers and mixtures enriched in one enantiomer, as well as diastereomers when there are two chiral centers, and mixtures partially enriched with specific diastereomers are within the scope of the present invention. It will be further appreciated by those skilled in the art that the present invention includes all the individual stereoisomers (e.g. enantiomers) , racemic mixtures or partially resolved mixtures of the compounds of formula I and, where appropriate, the individual tautomeric forms thereof.

In other words, in some embodiments, the present invention provides compounds of various stereoisomeric purities, i.e., diastereomeric or enantiomeric purity, with various "ee" or "de. " In some embodiments, the compound of formula I (e.g., as described herein) has an enantiomeric purity of at least 80％ee (e.g., 80％, 85％, 90％, 91％, 92％, 93％, 94％, 95％, 96％, 97％, 98％, 99％, 99.5％, 99.9％ee, or any ranges between those enumerated values) . In some embodiments, the compound of formula I (e.g., as described herein) has an enantiomeric purity of greater than 99.9％ee. In some embodiments, the compound of formula I (e.g., as described herein) has an enantiomeric purity of less than 80％ee. In some embodiments, the compound of formula I (e.g., as described herein) has a diastereomeric purity of at least 80％de (e.g., 80％, 85％, 90％, 91％, 92％, 93％, 94％, 95％, 96％, 97％, 98％, 99％, 99.5％, 99.9％de, or any ranges between those enumerated values) . In some embodiments, the compound of formula I (e.g., as described herein) has a diastereomeric purity of greater than 99.9％de. In some embodiments, the compound of formula I (e.g., as described herein) has a diastereomeric purity of less than 80％de.

The term “enantiomeric excess” or "ee" refers to a measure for how much of one enantiomer is present compared to the other. For a mixture of R and S enantiomers, the percent enantiomeric excess is defined as │R-S│*100, where R and S are the respective mole or weight fractions of enantiomers in a mixture such that R + S ＝ 1. With knowledge of the optical rotation of a chiral substance, the percent enantiomeric excess is defined as ( [α] _obs/ [α] _max) *100, where [α] _obs is the optical rotation of the mixture of enantiomers and [α] _max is the optical rotation of the pure enantiomer.

The term "diastereomeric excess" or "de" refers to a measure for how much of one diastereomer is present compared to the other and is defined by analogy to enantiomeric excess. Thus, for a mixture of diastereomers, D1 and D2, the percent diastereomeric excess is defined as │D1-D2│*100, where D1 and D2 are the respective mole or weight fractions of diastereomers in a mixture such that D1 + D2 ＝ 1.

The determination of diastereomeric and/or enantiomeric excess is possible using a variety of analytical techniques, including NMR spectroscopy, chiral column chromatography and/or optical polarimetry according to routine protocols will be familiar to those of ordinary skill in the art.

The racemates can be used as such or can be resolved into their individual isomers. The resolution can afford stereochemically pure compounds or mixtures enriched in one or more isomers. Methods for separation of isomers are well known (cf. Allinger N.L. and Eliel E.L. in "Topics in Stereochemistry" , Vol. 6, Wiley Interscience, 1971) and include physical methods such as chromatography using a chiral adsorbent. Individual isomers can be prepared in chiral form from chiral precursors. Alternatively individual isomers can be separated chemically from a mixture by forming diastereomeric salts with a chiral acid, such as the individual enantiomers of 10-camphorsulfonic acid, camphoric acid, alpha-bromocamphoric acid, tartaric acid, diacetyltartaric acid, malic acid, pyrrolidone-5-carboxylic acid, and the like, fractionally crystallizing the salts, and then freeing one or both of the resolved bases, optionally repeating the process, so as obtain either or both substantially free of the other； i.e., in a form having an enantiomeric purity of at least 91％, 92％, 93％, 94％, 95％, 96％, 97％, 98％, 99％or 99.5％by weight of the desired stereoisomer. Alternatively the racemates can be covalently linked to a chiral compound (auxiliary) to produce diastereomers which can be separated by chromatography or by fractional crystallization after which time the chiral auxiliary is chemically removed to afford the pure enantiomers.

The invention includes also pharmaceutically acceptable salts of the compounds represented by Formula I, preferably of those described below and of the specific compounds exemplified herein, and methods using such salts.

A “pharmaceutically acceptable salt” is intended to mean a salt of a free acid or base of a compound represented by Formula I that is non-toxic, biologically tolerable, or otherwise biologically suitable for administration to the subject. See, generally, S.M. Berge, et al., “Pharmaceutical Salts” , J. Pharm. Sci., 1977, 66: 1-19, and Handbook of Pharmaceutical Salts, Properties, Selection, and Use, Stahl and Wermuth, Eds., Wiley-VCH and VHCA, Zurich, 2002.

“Pharmaceutically acceptable salts” include, but are not limited to, acid addition salts formed by the compound of formula I with an inorganic acid, such as hydrochloride, hydrobromide, carbonate, bicarbonate, phosphate, sulfate, sulfite, nitrate and the like； as well as with an organic acid, such as formate, acetate, trifluoroacetate, malate, maleate, fumarate, tartrate, succinate, citrate, lactate, methanesulfonate, p-toluenesulfonate, 2-hydroxyethylsulfonate, benzoate, salicylate, stearate, and salts with alkane-dicarboxylic acid of formula HOOC- (CH₂) _n-COOH where n is 0-4, and the like. Also, “pharmaceutically acceptable salts” include base addition salts formed by the compound of formula I carring an acidic moiety with pharmaceutically acceptable cations, for example, sodium, potassium, calcium, aluminum, lithium, and ammonium. The molar ratio of the compound of formula I to the acid or the cation in the obtained pharmaceutically acceptable salt includes, but is not limited to, 1: 1, 1: 2, 1: 3, and 1: 4.

In addition, if a compound described herein is obtained as an acid addition salt, the free base can be obtained by basifying a solution of the acid addition salt. Conversely, if the product is a free base, an acid addition salt, particularly a pharmaceutically acceptable acid addition salt, may be produced by dissolving the free base in a suitable solvent and treating the solution with an acid, in accordance with conventional procedures for preparing acid addition salts from base compounds. Those skilled in the art will recognize various synthetic methodologies that may be used without undue experimentation to prepare non-toxic pharmaceutically acceptable acid addition salts or base addition salts.

The term “solvates” means solvent addition forms that contain either stoichiometric or non stoichiometric amounts of solvent. Some compounds have a tendency to trap a fixed molar ratio of solvent molecules in the solid state, thus forming a solvate. If the solvent is water the solvate formed is a hydrate, when the solvent is alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more molecules of water with one molecule of the substances in which the water retains its molecular state as H₂O, such combination being able to form one or more hydrates, for example, hemihydrates, monohydrate and dihydrate.

As used herein the terms "group" , "radical" or "fragment" are synonymous and are intended to indicate functional groups or fragments of molecules attachable to other fragments of molecules.

The term “active ingredient” is used to indicate a chemical substance which has biological activity. In some embodiments, an “active ingredient” is a chemical substance having pharmaceutical utility.

The terms “treating” , “treat” or “treatment” of a disease or disorder refers to administering one or more pharmaceutical substances, especially at least one compound of formula I and/or at least one pharmaceutically acceptable salt thereof described herein to a subject that has the disease or disorder, or has a symptom of a disease or disorder, or has a predisposition toward a disease or disorder, with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve, or affect a disease or disorder, the symptoms of the disease or disorder, or the predisposition toward the disease or disorder. In some embodiments, the disease or disorder is cancer.

The terms “treating” , “contacting” and “reacting” when referring to a chemical reaction mean adding or mixing two or more reagents under appropriate conditions to produce the indicated and/or the desired product. It should be appreciated that the reaction which produces the indicated and/or the desired product may not necessarily result directly from the combination of two reagents which were initially added, i.e., there may be one or more intermediates which are produced in the mixture which ultimately lead to the formation of the indicated and/or the desired product.

The term "effective amount" means an amount or dose of a PI₃K-inhibiting agent sufficient to generally bring about a therapeutic benefit in patients in need of treatment for a disease or disorder mediated by PI₃K activity. Effective amounts or doses of the active ingredient of the present invention may be ascertained by routine methods such as modeling, dose escalation studies or clinical trials, and by taking into consideration routine factors, e.g., the mode or route of administration or drug delivery, the pharmacokinetics of the agent, the severity and course of the disease or disorder, the subject's previous or ongoing therapy, the subject's health status and response to drugs, and the judgment of the treating physician. An exemplary dose is in the range of from about 0.0001 to about 200 mg of active agent per kg of subject's body weight per day, preferably about 0.001 to 100 mg/kg/day, or about 0.01 to 35 mg/kg/day, or about 0.1 to 10 mg/kg daily in single or divided dosage units (e.g., BID, TID, QID) . For a 70-kg human, an illustrative range for a suitable dosage amount is from about 0.05 to about 7 g/day, or about 0.2 to about 5 g/day. Once improvement of the patient's disease or disorder has occurred, the dose may be adjusted for maintenance treatment. For example, the dosage or the frequency of administration, or both, may be reduced as a function of the symptoms, to a level at which the desired therapeutic effect is maintained. Of course, if symptoms have been alleviated to an appropriate level, treatment may cease. Patients may, however, require intermittent treatment on a long-term basis upon any recurrence of symptoms.

The term “inhibition” or “inhibiting” indicates a decrease in the baseline activity of a biological activity or process. “Inhibition of PI₃K activity” refers to a decrease in the activity of PI₃K as a direct or indirect response to the presence of at least one compound of formula I and/or at least one pharmaceutically acceptable salt described herein, relative to the activity of PI₃K in the absence of the at least one compound of formula I and/or the at least one pharmaceutically acceptable salt thereof. The decrease in activity may be due to the direct interaction of the at least one compound of formula I and/or at least one pharmaceutically acceptable salt described herein with PI₃K, or due to the interaction of the at least one compound of formula I and/or at least one pharmaceutically acceptable salt described herein, with one or more other factors that in turn affect PI₃K activity. For example, the presence of at least one compound of formula I and/or at least one pharmaceutically acceptable salt described herein, may decrease PI₃K activity by directly binding to the PI₃K, by causing (directly or indirectly) another factor to decrease PI₃K activity, or by (directly or indirectly) decreasing the amount of PI₃K present in the cell or organism.

The term “subject” as used herein means mammals and non-mammals. Mammals means any member of the mammalia class including, but not limited to, humans； non-human primates such as chimpanzees and other apes and monkey species； farm animals such as cattle, horses, sheep, goats, and swine； domestic animals such as rabbits, dogs, and cats； laboratory animals including rodents, such as rats, mice, and guinea pigs； and the like. Examples of non-mammals include, but are not limited to, birds, and the like. The term “subject” does not denote a particular age or sex. In some embodiments, the subject is a human.

The term “pharmaceutically acceptable” means that the substance following this term is useful in preparing a pharmaceutical composition and is generally safe, non-toxic, and neither biologically nor otherwise undesirable, especially for human pharmaceutical use.

The term “about” is used herein to mean approximately, in the region of, roughly, or around. When the term “about” is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. In general, the term “about” is used herein to modify a numerical value above and below the stated value by a variance of 20％.

Technical and scientific terms used herein and not specifically defined have the meaning commonly understood by those skilled in the art to which the present invention pertains.

Embodiments of the Invention

In one aspect, the present invention provides a compound of formula I:

or pharmaceutically acceptable salts thereof； and its solvates, racemic mixture, enantiomers, diasteromers, tautomers, wherein:

Z₁ and Z₃ are independently N or CH； provided that Z₁ and Z₃ are not N simultaneously；

Z₂ and Z₄ are independently N or C, in which at least one of Z₂ and Z₄ is N；

R₁ is selected from halo, -CN, C_3-6 cycloalkyl, vinyl, ethynyl, phenyl, and C_1-6 alkyl, wherein the C_1-6 alkyl is optionally substituted with one or more groups (e.g., 1, 2, or 3) independently selected from halo, -OH and C_1-6 alkoxyl；

R₂ is C_1-6 alkyl optionally substituted with one or more groups (e.g., 1, 2, or 3) independently selected from C_3-6 cycloalkyl, phenyl and 5-7 membered (e.g., 5, 6, or 7-membered) heteroaryl； or

R₂ is selected from phenyl, C_3-6 cycloalkyl, 4-8 membered heterocycle, and 5-10 membered heteroaryl, each of which is optionally substituted with one or more groups (e.g., 1, 2, or 3) independently selected from halo, -CN, C_1-6 alkyl, -SR^a, -S (O) _nR^b, and C_1-6 alkoxyl, wherein the C_1-6 alkyl and C_1-6 alkoxyl are each optionally substituted by one or more halo；

R₃ and R₄ are each independently selected from H, C_1-6 alkoxyl, and C_1-4 alkyl, wherein the C_1-4 alkyl is optionally substituted by one or more -OH；

R₅ is H or a C_1-4 alkyl；

or R₃, R₅ and the atoms they are attached to form a 4-6 membered monocyclic or bicyclic saturated or partially unsaturated heterocyclic ring optionally containing an additional 1-2 heteroatoms independently selected from N, O and S, which is optionally substituted by one or more groups independently selected from halo and C_1-6 alkyl, wherein the C_1-6 alkyl is optionally substituted by one or more halo；

R^a and R^b are each independently H or a C_1-6 alkyl；

m is 0, 1, or 2；

n is 1 or 2；

W is formula II

wherein:

X₁ is C (O) , X₂ is NR₇, and X₃ is NR₈, CHR₉；

X₁ is S (O) or S (O) ₂, X₂ is CH₂ or NR₁₀, and X₃ is CH₂； or

X₁ is C (O) , X₂ is CH₂, and X₃ is NH, S or O； or

W is formula II-6-aor formula II-6-b:

R₆ is H, -NH₂, or -NH (C_1-6 alkyl) ； and

R₇, R₈, R₉, and R₁₀ are each independently selected from H, C_1-6 alkyl, and C_3-6 cycloalkyl.

In some embodiments of the compound of formula I, m is 0 or 1, wherein if m is 1, R₁ is selected from halo, -CN, C_3-6 cycloalkyl, phenyl, and C_1-6 alkyl, wherein the C_1-6 alkyl is optionally substituted with one or more groups independently selected from halo, OH, and C_1-6 alkoxyl； R₂ is C_1-6 alkyl optionally substituted with one or more groups independently selected from cyclopropyl and pyridyl； or R₂ is selected from phenyl, cyclopropyl, cyclobutyl, 6-membered heterocycle, and 6-membered heteroaryl, each of which is optionally substituted with one or more groups independently selected from the group consisting of halo, CN, C_1-6 alkyl, -SR^a, -S (O) _nR^b, and C_1-6 alkoxyl, wherein the C_1-6 alkyl and C_1-6 alkoxyl are each optionally substituted by one or more halo； wherein R^a and R^b are each independently a C_1-6 alkyl； and R₃ and R₄ are each independently H or a C_1-4 alkyl； R₅ is H or a C_1-4 alkyl； or R₃, R₅ and the atoms they are attached to form a 4-or 5-membered monocyclic saturated or partially unsaturated heterocyclic ring (e.g.,

or

) , which is optionally substituted by one or more groups independently selected from halo and C_1-6 alkyl, wherein the C_1-6 alkyl is optionally substituted by one or more halo.

In some embodiments, m is 0 or 1.

In an embodiment of the compound of formula I, m is 1.

In an embodiment of the compound of formula I, m is 0.

In some embodiments, R₁ is selected from halo, -CN, C_3-6 cycloalkyl (e.g., cyclopropyl or cyclobutyl) , phenyl, and C_1-6 alkyl (e.g., Me or Et) , wherein the C_1-6 alkyl is optionally substituted with one or more groups independently selected from halo, OH, and C_1-6 alkoxyl (e.g., MeO or EtO) . In some embodiments, m is 1, and R₁ is selected from halo (e.g., F, Cl, or Br) , -CN, C_3-6 cycloalkyl (e.g., cyclopropyl or cyclobutyl) , phenyl, and C_1-6 alkyl (e.g., Me or Et) , wherein the C_1-6 alkyl is optionally substituted with one or more (e.g., 1, 2, or 3) groups independently selected from halo (e.g., F) , OH, and C_1-6 alkoxyl (e.g., MeO or EtO) .

In an embodiment of the compound of formula I, R₁ is selected from halo (e.g., F, Cl, Br) , -CN, methyl, trifluoromethyl, hydroxymethyl, and methoxymethyl. In some embodiments, R₁ is cyclopropyl.

In an embodiment of the compound of formula I, m is 1, and R₁ is selected from halo, -CN, methyl, trifluoromethyl, hydroxymethyl, and methoxymethyl. In some embodiments, m is 1 and R₁ is cyclopropyl.

In an embodiment of the compound of formula I, m is 1, and R₁ is F, Cl, or Br. In some embodiments, m is 1 and R₁ is F. In some embodiments, m is 1 and R₁ is Cl.

In an embodiment of the compound of formula I, R₂ is phenyl or 5-10 membered heteroaryl, each of which is optionally substituted with one or more groups independently selected from halo, -CN, C_1-6 alkyl, -SR^a, -S (O) _nR^b, and C_1-6 alkoxyl, wherein the C_1-6 alkyl and C_1-6 alkoxyl are each optionally substituted by one or more halo. R^a and R^b are as defined herein.

In some embodiments of the compound of formula I, R₂ is C_1-6 alkyl (e.g., Me) optionally substituted with one or more (e.g., 1, 2, or 3) groups independently selected from cyclopropyl and pyridyl (e.g., pyrid-2-yl, pyrid-3-yl, or pyrid-4-yl) ； or R₂ is selected from phenyl, cyclopropyl, cyclobutyl, 6-membered heterocycle (e.g., piperidine, tetrahydropyran) , and 6-membered heteroaryl (e.g., pyridine) , each of which is optionally substituted with one or more (e.g., 1, 2, or 3) groups independently selected from the group consisting of halo (e.g., F, Cl) , CN, C_1-6 alkyl, -SR^a, -S (O) _nR^b, and C_1-6 alkoxyl, wherein the C_1-6 alkyl and C_1-6 alkoxyl are each optionally substituted by one or more halo； wherein R^a and R^b are each independently a C_1-6 alkyl.

In an embodiment of the compound of formula I, R₂ is phenyl optionally substituted with one or more groups independently selected from halo, -CN, C_1-6 alkyl, -SR^a, -S (O) _nR^b, and C_1-6 alkoxyl, wherein the C_1-6 alkyl and C_1-6 alkoxyl are each optionally substituted by one or more halo； R^a and R^b are each independently C_1-6 alkyl. In some embodiments, R₂ is phenyl, which is optionally substituted by one or more (e.g., 1, 2, or 3) groups independently selected from F, Cl, CN, CF₃, OMe, OCF₃, OCF₂H, -SMe, -S (O) Me and -S (O) ₂Me. In some embodiments, R₂ is phenyl. In some embodiments, R₂ is phenyl, which is substituted by one group selected from F, Cl, CN, CF₃, OMe, OCF₃, OCF₂H, -SMe, -S (O) Me and -S (O) ₂Me, e.g., at the 2-, 3-, or 4-position.

In an embodiment of the compound of formula I, R₂ is phenyl optionally substituted with one or more halo (e.g., F, Cl, or Br) . In some embodiments, R₂ is phenyl optionally substituted with one halo (e.g., F, Cl, or Br) , e.g., at the 2-, 3-, or 4-position.

In an embodiment of the compound of formula I, R₂ is a 6-membered saturated heterocycle containing one N or O, which is optionally substituted by a C_1-6 alkyl (e.g., Me) . In some embodiments, R₂ is a 6-membered saturated heterocycle containing one N. In some embodiments, the C_1-6 alkyl (e.g., Me) substituent is directed bonded with the N.

In an embodiment of the compound of formula I, R₂ is pyridyl. Preferably, R₂ is pyrid-2-yl.

In an embodiment of the compound of formula I, R₃ and R₄ are each independently H or a C_1-4 alkyl； R₅ is H or a C_1-4 alkyl； or R₃, R₅ and the atoms they are attached to form a 4-or 5-membered monocyclic saturated or partially unsaturated heterocyclic ring, which is optionally substituted by one or more groups independently selected from halo (e.g., F) and C_1-6 alkyl (e.g., Me or Et) , wherein the C_1-6 alkyl is optionally substituted by one or more halo (e.g., F) . In some embodiments, one of R₃ and R₄ is H, and the other of R₃ and R₄ is a C_1-4 alkyl (e.g., Me) . In some embodiments, R₅ is H. In some embodiments, R₃, R₅ and the atoms they are attached to form a 4-or 5-membered monocyclic saturated ring, which is optionally substituted by one or more groups independently selected from halo and C_1-6 alkyl, wherein the C_1-6 alkyl is optionally substituted by one or more halo.

In an embodiment of the compound of formula I, R₃, R₅ and the atoms they are attached to form

which is optionally substituted by one or more (e.g., 1 or 2) groups independently selected from halo and C_1-6 alkyl, wherein the C_1-6 alkyl is optionally substituted by one or more halo. In some embodiments, the

is not substituted.

which is optionally substituted by one or more (e.g., 1 or 2) groups independently selected from halo (e.g., F) , methyl, ethyl, and trifluoromethyl.

In some embodiments, the

is substituted by one group selected from halo (e.g., F) , methyl, ethyl, and trifluoromethyl, e.g., at the carbon atom not adjacent to the N atom. In some embodiments, the

is substituted by two groups independently selected from halo (e.g., F) , methyl, ethyl, and trifluoromethyl. In some embodiments, the two substituents are attached to the same carbon of the

In some embodiments, compounds having the

which is optionally substituted by one or two groups independently selected from halo (e.g., F) , methyl, ethyl, and trifluoromethyl, are characterized by having (1) an enantiomeric purity of at least 80％ee (e.g., 80％, 85％, 90％, 91％, 92％, 93％, 94％, 95％, 96％, 97％, 98％, 99％, 99.5％, 99.9％ee, or any ranges between those enumerated values) ； and/or (2) diastereomeric purity of at least 80％de (e.g., 80％, 85％, 90％, 91％, 92％, 93％, 94％, 95％, 96％, 97％, 98％, 99％, 99.5％, 99.9％de, or any ranges between those enumerated values) .

which is optionally substituted by one or more groups independently selected from halo and C_1-6 alkyl, wherein the C_1-6 alkyl is optionally substituted by one or more halo. In some embodiments, the

is not substituted.

which is optionally substituted by one or more (e.g., 1, 2, or 3) halo (e.g., F) . In some embodiments, compounds having the

which is optionally substituted by one or more (e.g., 1, 2, or 3) halo (e.g., F) , are characterized by having (1) an enantiomeric purity of at least 80％ee (e.g., 80％, 85％, 90％, 91％, 92％, 93％, 94％, 95％, 96％, 97％, 98％, 99％, 99.5％, 99.9％ee, or any ranges between those enumerated values) ； and/or (2) diastereomeric purity of at least 80％de (e.g., 80％, 85％, 90％, 91％, 92％, 93％, 94％, 95％, 96％, 97％, 98％, 99％, 99.5％, 99.9％de, or any ranges between those enumerated values) .

is not substituted.

which is optionally substituted by one or more (e.g., 1 or 2) groups independently selected from methyl and ethyl.

In an embodiment of the compound of formula I, R₃ and R₄ are each independently selected from H, C_1-4 alkyl, and C_1-6 alkoxyl； R⁵ is H.

In any one of the embodiments described herein (e.g., any of the preceding embodiments) , W is selected from

In any one of the embodiments described herein (e.g., any of the preceding embodiments) , R₆ is selected from H, -NH₂, and -NH-methyl.

In any one of the embodiments described herein (e.g., any of the preceding embodiments) , R₆ is -NH₂.

In any one of the embodiments described herein (e.g., any of the preceding embodiments) , R₇, R₈, R₉, and R₁₀ are independently H, methyl, ethyl, t-butyl or cyclopropyl.

In any one of the embodiments described herein (e.g., any of the preceding embodiments) , R₇, R₈, R₉, and R₁₀ are each H.

In some embodiments of the compound of formula I, when W is selected from II-1, II-2, II-3, and II-4, R₆ is selected from H, -NH₂, and -NH-methyl. Preferably, R₆ is -NH₂.

In some embodiments of the compound of formula I, when W is selected from II-1, II-2, II-3, and II-4, R₇, R₈, and R₉ are independently selected from H, methyl, ethyl, and cyclopropyl. Preferably, R₇, R₈, and R₉ are each H.

In any one of the preceding embodiments, W is

R₆ is -NH₂, and R₇ and R₉ are each H.

In some embodiments of the compound of formula I, when W is selected from II-1, II-2, II-3, and II-4, formula I is selected from I-1, I-2, I-3, I-4, and I-5,

wherein, R₁, R₂, R₃, R₄, R₅, W, and m are as defined in any one of the above embodiments. In some embodiments, m is 0 or 1.

Preferably, formula I is formula I-1. More preferably, formula I-1 is

wherein, R₁, R₂, R₃, R₄, R₅, and W are defined as in any one of the embodiments described herein (e.g., any of the above embodiments) .

In any one of the embodiments described herein (e.g., any of the preceding embodiments) , W is selected from II-5, II-6, and II-7,

In some embodiments of the compound of formula I, when W is selected from II-5, II-6, and II-7, R₆ is selected from H, -NH₂, and -NH-methyl. Preferably, R₆ is -NH₂.

In some embodiments of the compound of formula I, when W is selected from II-7, R₁₀ is selected from H and t-butyl.

In some embodiments of the compound of formula I, when W is selected from II-5, II-6, and II-7, formula I is I-1 or I-3,

wherein, R₁, R₂, R₃, R₄, R₅, W, and m are defined as in any one of the embodiments described herein (e.g., any of the above embodiments) . Preferably, formula I is formula I-1. More preferably, formula I-1 is

In any one of the preceding embodiments, W is II-8, II-9, or II-10,

In some embodiments of the compound of formula I, when W is II-8, II-9, or II-10, R₆ is selected from H, and -NH₂. Preferably, R₆ is -NH₂.

In some embodiments of the compound of formula I, when W is II-8, II-9, or II-10, formula I is

wherein, R₁, R₂, R₃, R₄, R₅, W, and m are defined as in any one of the embodiments described herein (e.g., any of the above embodiments) . Preferably, formula I-1 is

In some embodiments, it is to be understood by those of ordinary skill in the art that the compound of formula I having a W of II-8, II-9 or II-10 can exist either in a keto-form

a respective enol-form

or any equilibrium mixture thereof.

In a specific embodiment, the compound of formula I is selected from compounds numbered 1-232, as prepared in the examples. In some embodiments, the compound of formula I is selected from compounds numbered 16, 17, 40, 72, 104, and 145 as prepared in the examples.

In another aspect, the present invention provides a pharmaceutical composition, comprising at least one compound of formula I (e.g., any of the compounds described herein) and/or at least one pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient (e.g., a pharmaceutically acceptable carrier) .

In another aspect, the present invention provides a method of in vivo or in vitro inhibiting the activity of PI₃K, comprising contacting the PI₃K with an effective amount of at least one compound of formula I (e.g., any of the compounds described herein) and/or at least one pharmaceutically acceptable salt thereof.

In another aspect, the present invention provides a method of treating a disease responsive to inhibition of PI₃K in a subject, comprising administering to the subject in need thereof a therapeutically effective amount of at least one compound of formula I (e.g., any of the compounds described herein) and/or at least one pharmaceutically acceptable salt thereof.

In another aspect, the present invention provides a use of of at least one compound of formula I (e.g., any of the compounds described herein) and/or at least one pharmaceutically acceptable salt thereof described herein for treating a disease responsive to inhibition of PI₃K.

In another aspect, the present invention provides a use of at least one compound of formula I (e.g., any of the compounds described herein) and/or at least one pharmaceutically acceptable salt thereof described herein in the manufacture of a medicament for treating a disease responsive to inhibition of PI₃K.

In some embodiments, said disease responsive to inhibition of PI₃K is immune-based disease or cancer.

In some embodiments, said immune-based disease is rheumatoid arthritis, chronic obstructive pulmonary disease (COPD) , multiple sclerosis, asthma, glomerulonephritis, lupus, psoriasis, or inflammation related to any of the aforementioned； said cancer is lymphoma, acute myeloid leukemia, multiple myeloma or chronic lymphocytic leukemia.

General Synthetic Methods of the Invention

The compound of formula I described herein and/or a pharmaceutically acceptable salt thereof described herein can be synthesized from commercially available starting materials by methods well known in the art, taken together with the disclosure in this patent application. The following schemes illustrate general methods for preparation of the compounds disclosed herein. In each of the schemes, R¹, R², R³, R⁴, R⁵ and W are as defined herein.

Scheme I

Scheme II

Scheme III

Scheme IV

Scheme V

The compounds thus obtained can be further modified at their peripheral positions to provide the desired compounds. Synthetic chemistry transformations are described, for example, in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989) ； T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 3^rd Ed., John Wiley and Sons (1999) ； L. Fieser and M. Fieser, Fieser and Fieser’s Reagents for Organic Synthesis, John Wiley and Sons (1994) ； and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995) and subsequent editions thereof.

Before use, the compound of formula I and/or a pharmaceutically acceptable salt thereof described herein can be purified by column chromatography, high performance liquid chromatography, crystallization or other suitable methods.

Pharmaceutical Compositions and Utility

The compound of formula I (e.g., any of those described herein) and/or a pharmaceutically acceptable salt thereof described herein is used, alone or in combination with one or more additional active ingredients, to formulate pharmaceutical compositions of the invention. A pharmaceutical composition of the invention comprises: (a) an effective amount of at least one compound of formula I and/or a pharmaceutically acceptable salt thereof described herein； and (b) at least one pharmaceutically acceptable excipient (e.g., a pharmaceutically acceptable carrier) .

A pharmaceutically acceptable carrier refers to a carrier that is compatible with active ingredients of the composition (and in some embodiments, capable of stabilizing the active ingredients) and not deleterious to the subject to be treated. For example, solubilizing agents, such as cyclodextrins (which form specific, more soluble complexes with the the compound of formula I and/or a pharmaceutically acceptable salt thereof described herein) , can be utilized as pharmaceutical excipients for delivery of the active ingredients. Examples of other carriers include colloidal silicon dioxide, magnesium stearate, cellulose, sodium lauryl sulfate, and pigments such as D&C Yellow #10.

A composition comprising at least one compounds of formula I (e.g., any of those described herein) and/or at least one pharmaceutically acceptable salt thereof described herein can be administered in various known manners, such as orally, parenterally, rectally, topically, ocularly, by inhalation spray, or via an implanted reservoir. The term “parenteral” as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.

A composition described herein can be prepared in the form of tablet, capsule, sachet, dragee, powder, granule, lozenge, powder for reconstitution, liquid preparation, or suppository. Preferably, a composition is formulated for intravenous infusion, topical administration, or oral administration.

An oral composition can be any orally acceptable dosage form including, but not limited to, tablets, capsules, emulsions, and aqueous suspensions, dispersions and solutions. Commonly used carriers for tablets include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added to tablets. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions or emulsions are administered orally, the active ingredient can be suspended or dissolved in an oily phase combined with emulsifying or suspending agents. If desired, certain sweetening, flavoring, or coloring agents can be added.

A sterile injectable composition (e.g., aqueous or oleaginous suspension) can be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable Intermediate can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1, 3-butanediol. Among the pharmaceutically acceptable vehicles and solvents that can be employed are mannitol, water, Ringer’s solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium (e.g., synthetic mono-or di-glycerides) . Fatty acids, such as oleic acid and its glyceride derivatives are useful in the Intermediate of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions can also contain a long-chain alcohol diluent or dispersant, or carboxymethyl cellulose or similar dispersing agents.

An inhalation composition can be prepared according to techniques well known in the art of pharmaceutical formulation and can be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art.

A topical composition can be formulated in form of oil, cream, lotion, ointment, and the like, in which the active ingredients may be mixed with a pharmaceutical carrier at a concentration of about 0.1％to about 10％of drug to vehicle. Suitable carriers for the composition include vegetable or mineral oils, white petrolatum (white soft paraffin) , branched chain fats or oils, animal fats and high molecular weight alcohols (i.e., greater than C12) . In some embodiments, the pharmaceutically acceptable carrier is one in which the active ingredient is soluble. Emulsifiers, stabilizers, humectants and antioxidants may also be included as well as agents imparting color or fragrance, if desired. Additionally, transdermal penetration enhancers may be employed in those topical formulations. Examples of such enhancers can be found in U.S. Patents 3,989,816 and 4,444,762.

Creams may be formulated from a mixture of mineral oil, self-emulsifying beeswax and water in which mixture the active ingredient, dissolved in a small amount of an oil, such as almond oil, is admixed. An example of such a cream is one which includes, by weight, about 40 parts water, about 20 parts beeswax, about 40 parts mineral oil and about 1 part almond oil. Ointments may be formulated by mixing a solution of the active ingredient in a vegetable oil, such as almond oil, with warm soft paraffin and allowing the mixture to cool. An example of such an ointment is one which includes about 30％by weight almond oil and about 70％by weight white soft paraffin.

Suitable in vitro assays can be used to preliminarily evaluate the efficacy of the compound of formula I and/or a pharmaceutically acceptable salt thereof described herein, in inhibiting the activity of PI₃K kinase. The compound of formula I and/or a pharmaceutically acceptable salt thereof described herein can further be examined for efficacy in treating cancer, inflammatory or autoimmune disease by in vivo assays. For example, the compound of formula I and/or a pharmaceutically acceptable salt thereof described herein can be administered to an animal (e.g., a mouse model) having cancer, inflammatory or autoimmune disease and its therapeutic effects can be assessed. Based on the results, an appropriate dosage range and administration route for animals, such as humans, can also be determined.

The compound of formula I (e.g., any of those described herein) and/or a pharmaceutically acceptable salt thereof described herein can be used to achieve a beneficial therapeutic or prophylactic effect, for example, in subjects with cancer.

As used herein, the term "cancer" refers to a cellular disorder characterized by uncontrolled or disregulated cell proliferation, decreased cellular differentiation, inappropriate ability to invade surrounding tissue, and/or ability to establish new growth at ectopic sites. The term "cancer" includes, but is not limited to, solid tumors and bloodborne tumors. The term "cancer" encompasses diseases of skin, tissues, organs, bone, cartilage, blood, and vessels. The term "cancer" further encompasses primary and metastatic cancers.

Non-limiting examples of solid tumors include pancreatic cancer； bladder cancer； colorectal cancer； breast cancer, including metastatic breast cancer； prostate cancer, including androgen-dependent and androgen-independent prostate cancer； renal cancer, including, e.g., metastatic renal cell carcinoma； hepatocellular cancer； lung cancer, including, e.g., non-small cell lung cancer (NSCLC) , bronchioloalveolar carcinoma (BAC) , and adenocarcinoma of the lung； ovarian cancer, including, e.g., progressive epithelial or primary peritoneal cancer； cervical cancer； gastric cancer； esophageal cancer； head and neck cancer, including, e.g., squamous cell carcinoma of the head and neck； skin cancer, including e.g., malignant melanoma； neuroendocrine cancer, including metastatic neuroendocrine tumors； brain tumors, including, e.g., glioma, anaplastic oligodendroglioma, adult glioblastoma multiforme, and adult anaplastic astrocytoma； bone cancer； soft tissue sarcoma； and thyroid carcinoma.

Non-limiting examples of hematologic malignancies include acute myeloid leukemia (AML) ； chronic myelogenous leukemia (CML) , including accelerated CML and CML blast phase (CML-BP) ； acute lymphoblastic leukemia (ALL) ； chronic lymphocytic leukemia (CLL) ； Hodgkin's lymphoma； non-Hodgkin's lymphoma (NHL) , including follicular lymphoma and mantle cell lymphoma； B-cell lymphoma； T-cell lymphoma； multiple myeloma (MM) ； Waldenstrom's macroglobulinemia； myelodysplastic syndromes (MDS) , including refractory anemia (RA) , refractory anemia with ringed siderblasts (RARS) , refractory anemia with excess blasts (RAEB) , and RAEB in transformation (RAEB-T) ； and myeloproliferative syndromes.

The compound of formula I (e.g., any of those described herein) and/or a pharmaceutically acceptable salt thereof described herein can be used to achieve a beneficial therapeutic or prophylactic effect, for example, in subjects with an inflammatory disease.

The term “inflammatory disease” refers to pathological states resulting in inflammation, typically caused by neutrophil chemotaxis. Examples of such diseases include inflammatory skin diseases including psoriasis and atopic dermatitis； systemic scleroderma and sclerosis； responses associated with inflammatory bowel disease (IBD) (such as Crohn's disease and ulcerative colitis) ； ischemic reperfusion disorders including surgical tissue reperfusion injury, myocardial ischemic conditions such as myocardial infarction, cardiac arrest, reperfusion after cardiac surgery and constriction after percutaneous transluminal coronary angioplasty, stroke, and abdominal aortic aneurysms； cerebral edema secondary to stroke； cranial trauma, hypovolemic shock； asphyxia； adult respiratory distress syndrome； acute-lung injury； Behcet's Disease； dermatomyositis； polymyositis； multiple sclerosis (MS) ； dermatitis； meningitis； encephalitis； uveitis； osteoarthritis； lupus nephritis； autoimmune diseases such as rheumatoid arthritis (RA) , Sjorgen's syndrome, vasculitis； diseases involving leukocyte diapedesis； central nervous system (CNS) inflammatory disorder, multiple organ injury syndrome secondary to septicaemia or trauma； alcoholic hepatitis； bacterial pneumonia； antigen-antibody complex mediated diseases including glomerulonephritis； sepsis； sarcoidosis； immunopathologic responses to tissue/organ transplantation； inflammations of the lung, including pleurisy, alveolitis, vasculitis, pneumonia, chronic bronchitis, bronchiectasis, diffuse panbronchiolitis, hypersensitivity pneumonitis, idiopathic pulmonary fibrosis (IPF) , and cystic fibrosis； etc. The preferred indications include, without limitation, chronic inflammation, autoimmune diabetes, rheumatoid arthritis (RA) , rheumatoid spondylitis, gouty arthritis and other arthritic conditions, multiple sclerosis (MS) , asthma, systemic lupus erythrematosus, adult respiratory distress syndrome, Behcet's disease, psoriasis, chronic pulmonary inflammatory disease, graft versus host reaction, Crohn's Disease, ulcerative colitis, inflammatory bowel disease (IBD) , Alzheimer's disease, and pyresis.

The compound of formula I (e.g., any of those described herein) and/or a pharmaceutically acceptable salt thereof described herein can be used to achieve a beneficial therapeutic or prophylactic effect, for example, in subjects with an autoimmune disease.

The term “autoimmune disease” refers to a disease or disorder arising from and/or directed against an individual's own tissues or organs, or a co-segregate or manifestation thereof, or resulting condition therefrom. Examples of autoimmune diseases include, but are not limited to, chronic obstructive pulmonary disease (COPD) , allergic rhinitis, lupus, myasthenia gravis, multiple sclerosis (MS) , rheumatoid arthritis (RA) , psoriasis, inflammatory bowel disease (IBD) , asthma and idiopathic thrombocytopenic purpura, and myeloproliferative disease, such as myelofibrosis, PV /ET (Post-Polycythemia Vera/Essential Thrombocythemia Myelofibrosis) .

In addition, the compound of formula I (e.g., any of those described herein) and/or a pharmaceutically acceptable salt thereof described herein may be used in combination with additional active ingredients in the treatment of cancer, inflammatory or autoimmune disease. The additional active ingredients may be coadministered separately with the compound of formula I and/or a pharmaceutically acceptable salt thereof described herein or included with such an ingredient in a pharmaceutical composition according to the invention. In an exemplary embodiment, additional active ingredients are those that are known or discovered to be effective in the treatment of diseases mediated by PI₃K activity, such as another PI₃K modulator or a compound active against another target associated with the particular disease. The combination may serve to increase efficacy (e.g., by including in the combination a compound potentiating the potency or effectiveness of the compound of formula I and/or a pharmaceutically acceptable salt thereof described herein) , decrease one or more side effects, or decrease the required dose of the compound of formula I and/or a pharmaceutically acceptable salt thereof described herein.

EXAMPLES

The examples below are intended to be purely exemplary and should not be considered to be limiting in any way. Efforts have been made to ensure accuracy with respect to numbers used (for example, amounts, temperature, etc. ) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric. All MS data were checked by agilent 6120 or agilent 1100. All NMR data were generated using a Varian 400-MR machine. All reagents, except intermediates, used in this invention are commercially available. All compound names except the reagents were generated by Chemdraw 12.0.

In the following examples, the abbreviations below are used:

Molecular sieves

aq. aqueous solution

ADP Adenosine diphosphate

ATP Adenosine triphosphate

n-BuOH n-butanol

BOP benzotriazol-1-yloxytris (dimethylamino) -phosphonium hexafluorophosphate

calcd. calculated

CHAPS 3- [ (3- Cholamidopropyl) dimethylammonio] propanesulfonate

conc. concentrated

DAST diethylaminosulfur trifluoride

dba dibenzylideneacetone

DBU 1, 8-diazabicyclo [5.4.0] undec-7-ene

DCM dichloromethane

DHP 3, 4-dihydro-2H-pyran

DIEA N, N-diisopropylethylamine

DIBAL-H Diisobutylaluminum hydride

DMA N, N-dimethylacetamide

DMF N, N-dimethylformamide

DPPA diphenylphosphoryl azide

dppf 1, 1' -bis (diphenylphosphino) ferrocene

DTT DL-Dithiothreitol

Eaton's reagent 7.7 wt％phosphorus pentoxide solution in methanesulfonic acid

EDC 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride

EGTA Glycol-bis- (2-aminoethylether) -N, N, N' , N' -tetraacetic acid

EtOAc ethyl acetate

g gram (s)

h hour (s)

HATU 2- (1H-7-azabenzotriazol-1-yl) --1, 1, 3, 3-tetramethyl uronium hexafluorophosphate methanaminium

HBTU 2- (1H-Benzotriazole-1-yl) -1, 1, 3, 3-Tetramethyluronium hexafluorophosphate

HEPES 4- (2-Hydroxyethyl) -1-piperazineethanesulfonic acid

m-CPBA 3-chloroperoxybenzoic acid

MeOH methanol

mg milligram (s)

min minute (s)

mL milliliter (s)

NCS N-chlorosuccinimide

PE petroleum ether

PyBrOP Bromo-tris-pyrrolidinophosphoniumhexafluorophosphate

PCC Pyridinium Chlorochromate

rt. room temperature

Selectfluor 1-chloromethyl-4-fluoro-1, 4-diazoniabicyclo [2.2.2] octane bis (tetrafluoroborate)

SEM 2- (trimethylsilyl) ethoxymethyl

TBAF tetrabutylammonium fluoride

TBSCl t-butylchlorodimethylsilane

TEA triethylamine

TFA trifluoroacetic acid

THF tetrahydrofuran

THP tetrahydropyran

TLC thin-layer chromatography

TMS trimethylsilyl

TsOH p-toluenesulfonic acid

TsCl p-toluenesulfonic chloride

Xantphos 4, 5-Bis (diphenylphosphino) -9, 9-dimethylxanthene

Intermediate I-1: 4-Chloro-2- (methylthio) -6, 7-dihydrothieno [3, 2-d] pyrimidine

Step 1. 2- (Methylthio) -6, 7-dihydrothieno [3, 2-d] pyrimidin-4-ol

Methyl 3-oxotetrahydrothiophene-2-carboxylate (3.2 g, 20 mmol) , methyl carbamimidothioate 0.5H₂SO₄ (2.78 g, 20 mmol) and Na₂CO₃ (2.2 g, 20 mmol) were dissolved in H₂O (30 mL) , the reaction mixture was stirred at rt. overnight. The resulting precipitate was filtered to afford the title compound as a solid (1.6 g) . Yield: 40％. MS (ESI) : calcd. value for C₇H₈N₂OS₂ is 200.0, m/z measured value is 200.9 (M+1) ⁺.

Step 2. 4-Chloro-2- (methylthio) -6, 7-dihydrothieno [3, 2-d] pyrimidine

To the solution of 2- (methylthio) -6, 7-dihydrothieno [3, 2-d] pyrimidin-4-ol (1 g, 5 mmol) in CH₃CN (30 mL) was added POCl₃ (5 mL) . The resulting mixture was stirred at 90℃ for 3 h, and then cooled to rt., and poured into water. The resulting precipitate was filtered to afford the title compound as a solid (930 mg) . Yield: 40％. MS (ESI) : calcd. value for C₇H₇ClN₂S₂ is 218.0, m/z measured value is 218.8 (M+1) ⁺.

Intermediate I-2: tert-Butyl 2-amino-4- (methylthio) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) -carboxylate

Step 1. tert-Butyl 3- (bis (methylthio) methylene) -2, 4-dioxopyrrolidine-1-carboxylate

K₂CO₃ (2.7 g, 20 mmol) was added to tert-butyl 2, 4-dioxopyrrolidine-1-carboxylate (2 g, 10 mmol) in dry DMF (60 mL) , the resulting mixture was stirred at rt. for 0.5 h, then carbon disulfide (1.52 g, 20 mmol) was added, the mixture was stirred at rt. for 2 h, and then CH₃I (4.2 g, 30 mmol) in dry DMF (5 mL) was added dropwise, the mixture was stirred at rt. for another 1h. After the reaction was completed, aq. Na₂S₂O₅ was added to quench the reaction. The mixture was extracted by EtOAc, the organic layers were dried over anhydrous Na₂SO₄, and concentrated to dryness to afford the crude title compound as an oil, which was used in next step directly. MS (ESI) : calcd. value for C₁₂H₁₇NO₄S₂ is 303.1, m/z measured value is 248.0 (M-56+1) ⁺.

Step 2. tert-Butyl 2-amino-4- (methylthio) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) -carboxylate

To a mixture of guanidine hydrochloride (1.43 g, 15 mmol) in DMF (15 mL) was added Et₃N (2.02 g, 20 mmol) , the resulting mixture was stirred at rt. for 0.5 h, and then the above crude product (3.03 g, 10 mmol) in DMF (40 mL) was added. The mixture was stirred at 70℃ overnight. After the reaction was completed, NH₄Cl solution was added and extracted by EtOAc, the organic layers were concentrated to dryness. The resulting residue was purified by flash column chromatography to afford the title compound as a solid (1.62 g) .Yield: 55％. ¹H NMR (400 MHz, CDCl₃) δ 5.51 (s, 2H) , 4.50 (s, 2H) , 2.50 (s, 3H) , 1.56 (s, 9H) . MS (ESI) : calcd. value for C₁₂H₁₆N₄O₃S is 296.4 , m/z measured value is 197.0 (M-100+1) ⁺.

Intermediate I-3: tert-Butyl 2-amino-4- (methylthio) -5-oxo-7, 8-dihydropyrido [4, 3-d] pyrimidine-6 (5H) -carboxylate

Step 1. tert-Butyl 3- (bis (methylthio) methylene) -2, 4-dioxopiperidine-1-carboxylate

To a solution of tert-butyl 2, 4-dioxopiperidine-1-carboxylate (4.2 g, 20 mmol) in DMF (15 mL) were added CS₂ (3 g, 40 mmol) and K₂CO₃ (5.5 g, 40 mmol) . The resulting mixture was stirred at rt. for 2 h, then CH₃I (5.7 g, 40 mmol ) in DMF (5 mL) was added dropwise, and stirred for another 1 h. After the reaction was completed, the mixture was poured into water, and extracted with EtOAc. The organic layers were washed with aq. Na₂S₂O₃ and brine, and then concentrated to dryness to afford the title compound as a solid (5 g) . Yield: 79％.

Step 2. tert-Butyl 2-amino-4- (methylthio) -5-oxo-7, 8-dihydropyrido [4, 3-d] pyrimidine-6 (5H) -carboxylate

To a suspension of guanidine hydrochloride (722 mg, 7.6 mmol) in DMF (20 mL) was added K₂CO₃ (3.1 g, 22.7 mmol) . The resulting mixture was stirred at rt. for 10 min, and then above solid (2.4 g, 7.6 mmol) was added. The mixture was stirred at 90℃ for 2 h, then poured into water, and extracted with EtOAc. The organic layers were washed with water and brine, and then concentrated to dryness to afford the title compound as a solid (1.6 g) . Yield: 68％. MS (ESI) : calcd. value for C₁₃H₁₈N₄O₃S is 310.11, m/z measured value is 310.8 (M+1) ⁺.

Intermediate I-4: 4-Chloro-6-methyl-2- (methylthio) -7, 8-dihydropyrido [4, 3-d] pyrimidin-5 (6H) -one

Step 1. 4-Amino-6-methyl-2- (methylthio) -7, 8-dihydropyrido [4, 3-d] pyrimidin-5 (6H) -one

4-Methoxy-1-methyl-2-oxo-1, 2, 5, 6-tetrahydropyridine-3-carbonitrile (1.55 g, 9.33 mmol) , S-methylisothiouronium sulfate (7.78 g, 28 mmol) and DBU (2.12 g, 14 mmol) were dissolved in MeOH (50 mL) . The resulting mixture was stirred at reflux for 48 h, and then concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (1.8 g) . Yield: 86％. MS (ESI) : calcd. value for C₉H₁₂N₄OS is 224.1, m/z measured value is 225.0 (M+1) ⁺.

Step 2. 4-Chloro-6-methyl-2- (methylthio) -7, 8-dihydropyrido [4, 3-d] pyrimidin-5 (6H) -one

4-Amino-6-methyl-2- (methylthio) -7, 8-dihydropyrido [4, 3-d] pyrimidin-5 (6H) -one (448 mg, 2 mmol) , tert-butyl nitrite (309 mg, 3 mmol) and CuCl₂ (322 mg, 2.4 mmol) were dissolved in CH₃CN (20 mL) , the resulting mixture was stirred at 80℃ for 2 days, and then concentrated to dryness. The residue was used for the next step without purification. MS (ESI) : calcd. value for C₉H₁₀ClN₃OS is 243.0, m/z measured value is 244.0 (M+1) ⁺.

Intermediate I-5: 4-Chloro-6-methyl-2- (methylthio) pyrido [4, 3-d] pyrimidin-5 (6H) -one

Step 1. Ethyl 4- ( (2, 4-dimethoxybenzyl) amino) -6-methyl-2- (methylthio) pyrimidine-5-carboxylate

Ethyl 4-chloro-6-methyl-2- (methylthio) pyrimidine-5-carboxylate (1.22 g, 5 mmol) , (2, 4-dimethoxyphenyl) methanamine (835 mg, 5 mmol) and Et₃N (1.4 mL) were dissolved in CH₃CN (50 mL) , the resulting mixture was stirred at rt. overnight, and then concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (1.8 g) . Yield: 95％. MS (ESI) : calcd. value for C₁₈H₂₃N₃O₄S is 377.1, m/z measured value is 378.1 (M+1) ⁺.

Step 2. Ethyl 4- ( (2, 4-dimethoxybenzyl) amino) -6- (2- (dimethylamino) vinyl) -2- (methylthio) pyrimidine-5-carboxylate

Ethyl 4- ( (2, 4-dimethoxybenzyl) amino) -6-methyl-2- (methylthio) pyrimidine-5-carboxylate (1.2 g, 3.18 mmol) and 1, 1-dimethoxy-N, N-dimethylmethanamine (378 mg, 3.18 mmol) were dissolved in DMF (10 mL) . The resulting mixture was stirred at 130℃overnight, and then concentrated to dryness to afford the crude title compound, which was used for next step without purification.

Step 3. 4- ( (2, 4-Dimethoxybenzyl) amino) -6-methyl-2- (methylthio) pyrido [4, 3-d] pyrimidin-5 (6H) -one

The above residue (1.375 g, 3.18 mmol) was dissolved in EtOH (50 mL) , and then methanamine (23％in EtOH, 10 mL) was added. The resulting mixture was stirred at 90℃overnight, and then concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (372 mg) . Yield: 31％. MS (ESI) : calcd. value for C₁₈H₂₀N₄O₃S is 372.1, m/z measured value is 373.3 (M+1) ⁺.

Step 4. 4-Amino-6-methyl-2- (methylthio) pyrido [4, 3-d] pyrimidin-5 (6H) -one

The mixture of 4- ( (2, 4-Dimethoxybenzyl) amino) -6-methyl-2- (methylthio) pyrido [4, 3-d] pyrimidin-5 (6H) -one (372 mg, 1 mmol) dissolved in TFA (3 mL) was stirred at reflux for 1 h, and then concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (120 mg) . Yield: 54％. MS (ESI) : calcd. value for C₉H₁₀N₄OS is 222.1, m/z measured value is 223.1 (M+1) ⁺.

Step 5. 4-Chloro-6-methyl-2- (methylthio) pyrido [4, 3-d] pyrimidin-5 (6H) -one

4-amino-6-methyl-2- (methylthio) pyrido [4, 3-d] pyrimidin-5 (6H) -one (120 mg, 0.54 mmol) , tert-butyl nitrite (83 mg, 0.81 mmol) and copper (II) chloride (86 mg, 0.65 mmol) were dissolved in CH₃CN (10 mL) , the mixture was stirred at reflux overnight. The resulting precipitate was filtered and the filtrate was purified by flash column chromatography to afford the title compound as a solid (48 mg) . Yield: 36.9％. MS (ESI) : calcd. value for C₉H₈ClN₃OS is 241.0, m/z measured value is 241.9 (M+1) ⁺.

Intermediate I-6: tert-Butyl 4- (ethylthio) -2- (methylthio) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) -carboxylate

Step 1. tert-Butyl 3- (bis (ethylthio) methylene) -2, 4-dioxopyrrolidine-1-carboxylate

To a solution of tert-butyl 2, 4-dioxopyrrolidine-1-carboxylate (4 g, 20 mmol) in dry DMF(60 mL) was added K₂CO₃ (5.8 g, 42 mmol) , followed by CS₂ (3.19 g, 42 mmol) . The resulting mixture was stirred at rt. for 2 h, and then iodoethane (6.55 g, 42 mmol) was added. The mixture was stirred at rt. overnight, and then diluted with EtOAc and water. The organic layer was separated and dried over anhydrous Na₂SO₄, and then concentrated to dryness to afford the crude title product as an oil. MS (ESI) : calcd. value for C₁₄H₂₁NO₄S₂ is 331.09, m/z measured value is 684.9 (2M+23) ⁺.

Step 2. tert-Butyl 4- (ethylthio) -2- (methylthio) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) -carboxylate

To a solution of above crude product in EtOH (100 mL) was added DIEA (7.74 g, 60 mmol) and 2-Methyl-2-thiopseudourea sulfate (2.78 g, 20 mmol) . The resulting mixture was stirred at 100℃ for 6h, and then concentrated to dryness. The residue was diluted with EtOAc and water. The organic layer was washed with brine, dried over anhydrous Na₂SO₄, and then concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (1.5 g) . Yield: 22％. MS (ESI) : calcd. value for C₁₄H₁₉N₃O₃S₂ is 341.09, m/z measured value is 704.9 (2M+23) ⁺。

Intermediate I-7: tert-Butyl 4- (methylthio) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) -carboxylate

To a solution of Intermediate I-2 (400 mg, 1.35 mmol) in THF (8 mL) was added tert-butyl nitrite (696 mg, 6.76 mmol) . The resulting mixture was stirred at 70℃ for 20 h, and then concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (170 mg) . Yield: 45％. MS (ESI) : calcd. value for C₁₂H₁₅N₃O₃S is 281.08, m/z measured value is 282.0 (M+1) ⁺.

Intermediate I-8: 4- (Ethylthio) -2- (methylthio) pyrido [4, 3-d] pyrimidin-5 (6H) -one

To a solution of ethyl 4-chloro-6-methyl-2- (methylthio) pyrimidine-5-carboxylate (1.23 g, 5 mmol) in DMF (15 mL) was added K₂CO₃ (1.36 g, 10 mmol) , followed by ethanethiol (620 mg, 10 mmol) . The resulting mixture was stirred at r.t. overnight, and then DMF-DMA (2 mL) was added. The mixture was stirred at 130℃ overnight, and then concentrated to dryness. The residue was dissolved in EtOH (10 mL) , and then NH₄OH (10 mL) was added, the mixture was stirred at reflux overnight, and then concentrated to dryness, the residue was purified by flash column chromatography to afford the title compound as a solid (250 mg) . Yield: 20％. MS (ESI) : calcd. value for C₁₀H₁₁N₃OS₂ is 253.0, m/z measured value is 254.0 (M+1) ⁺.

Intermediate I-9: tert-butyl 2-amino-4- (methylthio) -5-oxo-5H-pyrrolo [2, 3-d] pyrimidine-7 (6H) -carboxylate

Intermediate I-9 was isolated as a byproduct from the procedure of intermediate I-2. ¹H NMR (400 MHz, CDCl₃) δ 5.70 (s, 2H) , 4.13 (s, 2H) , 2.50 (s, 3H) , 1.54 (s, 9H) . MS (ESI) : calcd. value for C₁₂H₁₆N₄O₃S is 296.4, m/z measured value is 615.2 (2M+23) ⁺.

Intermediate I-10: 2-amino-6-methyl-4- (methylthio) -6, 7-dihydro-5H-pyrrolo [3, 4-d] pyrimidin-5-one

Step1: ethyl 2- (bis (methylthio) methylene) -3-oxobutanoate

To a solution of ethyl 3-oxobutanoate (1.3 g, 10 mmol) in dry DMF (20 mL) was added K₂CO₃ (2.9 g, 21 mmol) , followed by CS₂ (1.59 g, 21 mmol) , the resulting mixture was stirred at r.t. for 2 h, then added iodomethane (2.98 g, 21 mmol) . The resulting mixture was stirred at r.t. for another 2 h. The reaction mixture was used for next step without purification. MS (ESI) : calcd. value for C₉H₁₄O₃S₂ is 234.04, m/z measured value is 257.0 (M+23) ⁺.

Step2: ethyl 2-amino-4-methyl-6- (methylthio) pyrimidine-5-carboxylate

To the solution of the above reaction mixture was added guanidine hydrochloride (955 mg, 10 mmol) and K₂CO₃ (2.76 g, 20 mmol) , the resulting mixture was stirred at 90℃ overnight. After cooling to r.t., it was poured into ice-water, the precipitate was collected through filtration to afford the title compound as brown solid (1 g) . Yield: 44％. MS (ESI) : calcd. value for C₉H₁₃N₃O₂S is 227.07, m/z measured value is 228.0 (M+1) ⁺.

Step3: ethyl 2-amino-4- (bromomethyl) -6- (methylthio) pyrimidine-5-carboxylate

To a solution of ethyl 2-amino-4-methyl-6- (methylthio) pyrimidine-5-carboxylate (300 mg, 1.32 mmol) in chloroform (10 mL) was added Br₂ (423 mg, 2.64 mmol) and AcOH (2 drops) . The resulting mixture was stirred at 90℃ for 4 h. The resulting mixture was used for next step without purification. MS (ESI) : calcd. value for C₉H₁₂BrN₃O₂S is 304.98, m/z measured value is 307.9 (M+1) ⁺.

Step4: 2-amino-6-methyl-4- (methylthio) -6, 7-dihydro-5H-pyrrolo [3, 4-d] pyrimidin-5-one

To a mixture of the above step was added a solution of methanamine in EtOH (27％-32％, 1 mL) . The mixture was stirred at r.t. overnight. The organic solvent was evaporated and the residue was purified by flash column chromatography to afford the title compound as yellow solid (200 mg) . Yield: 72％. MS (ESI) : calcd. value for C₈H₁₀N₄OS is 210.06, m/z measured value is 211.0 (M+1) ⁺

Intermediate I-11: 5-amino-2- (tert-butyl) -7- (methylthio) -2, 3-dihydroisothiazolo [4, 5-d] pyrimidine 1, 1-dioxide

Step 1: sodium 2-ethoxy-2-oxoethane-1-sulfonate

A solution consisting of ethyl 2-chloroacetate (300 g, 2.45 mol) and EtOH (1.1 L) was added to a solution consisting of Na₂SO₃ (206 g, 1.63 mol) and H₂O (1 L) . The resultant mixture was stirred at reflux for 16 hours before concentrating to dryness under reduced pressure to afford the title compound (350 g, crude) as white solid, which was used in the next step without purification.

Step 2: ethyl 2- (chlorosulfonyl) acetate

Sodium 2-ethoxy-2-oxoethanesulfonate (300 g, 1.58 mol) and PCl₅ (360 g, 1.73 mol) were added to a 1 L round-bottomed flask. The resultant mixture was stirred for 30 minutes during which time there was an exotherm and the solids liquified. The resulting mixture was stirred and heated at 100 ℃ for 16 hours, then cooled to room-temperature and concentrating to dryness under reduced pressure. The residue was triturated with toluene (500 mL) , filtered, and the filtrate concentrated to dryness under reduced pressure to afford the title compound (180 g, crude) , which was used in the next step without purification.

Step 3: ethyl tert-butylglycinate

2-Methylpropan-2-amine (59.7 g, 816 mmol) was added drop-wise to a mixture consisting of ethyl 2-chloroacetate (100 g, 816 mmol) , K₂CO₃ (221 g, 1.60 mol) , MeCN (1 L) , and ethyl acetate (100 mL) . The resultant mixture was stirred at room-temperature for 16 hours before the suspension was isolated by filtration. The filtrate was concentrated to dryness under reduced pressure to afford the title product (115 g, crude) . ¹H NMR (400 MHz, CDCl₃) : 4.17 (q, J ＝ 7.1 Hz, 2H) , 3.37 (s, 2H) , 1.26 (t, J ＝ 7.1 Hz, 3H) , 1.09 (s, 9H) .

Step 4: ethyl N- (tert-butyl) -N- ( (2-ethoxy-2-oxoethyl) sulfonyl) glycinate

A solution consisting of ethyl 2- (chlorosulfonyl) acetate (176 g, 0.943 mol) and THF (0.5 L) was added drop-wise to a -20 ℃ (dry ice/acetone) solution consisting of ethyl 2-(tert-butylamino) acetate (150 g, 0.943 mol) , Et₃N (95.1 g, 0.942 mol) and THF (1.5 L) . The resultant mixture was stirred for 16 hours with gradual warming to room-temperature before it was diluted with ethyl acetate (1 L) and washed with saturated NH₄Cl (500 mL x 2) . The organic phase was dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness under reduced pressure to give the crude product, which was purified by fast column chromatography to afford the title compound (140 g, 48.0％) .

Step 5: Ethyl 2- (tert-butyl) -4-oxoisothiazolidine-5-carboxylate 1, 1-dioxide

Sodium (7.9 g, 0.34 mol) was added in portions to EtOH (200 mL) and the resultant mixture stirred at reflux until the sodium was completely consumed (15 minutes) . A solution of ethyl 2- (N- (tert-butyl) -2-ethoxy-2-oxoethylsulfonamido) acetate (53 g, 0.17 mol) and EtOH (150 mL) was added drop-wise to the NaOEt solution at 80 ℃. The resultant mixture was stirred and heated at 80 ℃ for 2 hours before cooling to room-temperature. The mixture was poured into saturated NH₄Cl (400 mL) , and extracted with DCM (300 mL x 5) . The combined organic extracts were dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness under reduced pressure to afford the title compound (26 g, crude) , which was used in the next step without purification.

Step 6: 2- (tert-Butyl) isothiazolidin-4-one 1, 1-dioxide

A solution consisting of KOH (120 g, 2.14 mol) and H₂O (300 mL) was added to a solution consisting of ethyl 2- (tert-butyl) -4-oxoisothiazolidine-5-carboxylate 1, 1-dioxide (13.0 g, 49.3 mmol) , and 1, 4-dioxane (400 mL) . The resultant mixture was stirred at reflux for 16 hours before cooling to room-temperature. The reaction mixture was acidified with concentrated HCl to pH ＝ 5 and extracted with ethyl acetate (100 mL x 3) . The combined organic extracts were dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness under reduced pressure to afford the title compound (4.0 g, 42％) . ¹H NMR (400 MHz, CDCl₃) : 3.91 (s, 2H) , 3.84 (s, 2H) , 1.46 (s, 9H) .

Step 7: 5- (Bis (methylthio) methylene) -2- (tert-butyl) isothiazolidin-4-one 1, 1-dioxide

Sodium hydride in mineral oil (209 mg, 60％purity, 5.23 mmol) was added in one portion to a 0 ℃ (ice/water) solution consisting of 2- (tert-butyl) isothiazolidin-4-one 1, 1-dioxide (500 mg, 2.61 mmol) , CS₂ (199 mg, 2.61 mmol) and DMF (5 mL) . The resultant mixture was stirred for 1 hour with gradual warming to room-temperature and then treated with MeI (742 mg, 5.23 mmol) at 0 ℃ under N₂. The resulting mixture was stirred at room-temperature for 16 hours before it was quenched with H₂O (20 mL) and extracted with ethyl acetate (20 mL x 3) . The combined organic extracts were dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness under reduced pressure to give the crude product, which was purified by FCC to afford the title compound (200 mg, 26.0％) . ¹H NMR (400 MHz, CDCl₃) : 3.78 (s, 2H) , 2.80 -2.60 (m, 6H) , 1.48 (s, 9H) .

Step 8: 5-Amino-2- (tert-butyl) -7- (methylthio) -2, 3-dihydroisothiazolo [4, 5-d] pyrimidine 1, 1-dioxide

Guanidine carbonate salt (903 mg, 5.01 mmol) was added to a mixture consisting of 5- (bis (methylthio) methylene) -2- (tert-butyl) isothiazolidin-4-one 1, 1-dioxide (1.20 g, 5.01 mmol) , K₂CO₃ (1.42 g, 10.3 mol) and MeCN (18 mL) . The resultant mixture was stirred and heated at 90 ℃ via microwave irradiation for 1 hour. The reaction mixture was cooled to room-temperature and the suspension was isolated by filtration. The filtrate was concentrated to dryness under reduced pressure to give the crude product, which was purified by FCC to afford the title compound (318 mg, 22.0％) . MS (ESI) : mass calcd. for C₁₀H₁₆N₄O₂S₂ 288.07, m/z found 289.1 [M+H] ⁺. ¹H NMR (400 MHz, DMSO-d₆) : 7.64 (s, 2H) , 4.28 (s, 2H) , 2.56 (s, 3H) , 1.41 (s, 9H) .

Intermediate I-12: tert-Butyl 2-amino-7-methyl-4- (methylthio) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) -carboxylate

Step 1: tert-butyl 3- (bis (methylthio) methylene) -5-methyl-2, 4-dioxopyrrolidine-1-carboxylate

CS₂ (24.5 g, 322 mmol) was added to a mixture consisting of tert-butyl 2-methyl-3, 5-dioxopyrrolidine-1-carboxylate (38.0 g, 160 mmol) , K₂CO₃ (55.4 g, 401 mmol) , and DMF (150 mL) . The resultant mixture was stirred at 20 ℃ for 2 hours and then treated with a solution consisting of iodomethane (62.1 g, 438 mmol) and DMF (50 mL) . The resultant mixture was stirred at 20 ℃ for 2 hours before being diluted with ethyl acetate (150 mL) and washed with brine (100 mL x 2) . The organic extract was dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness under reduced pressure to give the crude product, which was purified by FCC to afford the title compound (15 g, 27％) . ¹H NMR (400 MHz, CDCl₃) : 4.20 (q, J ＝ 6.5 Hz, 1H) , 2.65 (s, 6H) , 1.56 (s, 9H) , 1.48 (d, J ＝ 6.4 Hz, 3H) .

Step 2: tert-Butyl 2-amino-7-methyl-4- (methylthio) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) -carboxylate

K₂CO₃ (13 g, 94 mmol) was added to a mixture consisting of tert-butyl 3- (bis (methylthio) methylene) -5-methyl-2, 4-dioxopyrrolidine-1-carboxylate (15 g, 43 mmol) , guanidine hydrochloride (5.4 g, 57 mmol) , and acetontrile (100 mL) . The resultant mixture was stirred at 90 ℃ for 16 hours before being concentrated to dryness under reduced pressure. The residue was diluted with H₂O (100 mL) and extracted with dichloromethane (100 mL x 3) . The combined organic extracts were dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness under reduced pressure to give the crude product, which was purified by preparative HPLC. The pure fractions were collected and the volatiles were removed under vacuum. The residue was suspended in water (10 mL) , the mixture frozen using dry ice/acetone, and then lyophilized to dryness to afford the title compound (1.1 g) . Yield: 8％. MS (ESI) : mass calcd. for C₁₃H₁₈N₄O₃S 310.11_, m/z found 311.1 [M+H] ⁺. ¹H NMR (400 MHz, CDCl₃) : 5.43 (br.s., 2H) , 4.75 (q, J ＝ 6.8 Hz, 1H) , 2.52 (s, 3H) , 1.60 -1.57 (m, 12H) .

Intermediate I-12-A1 and Intermediate I-12-A2

Intermediate I-12-A1: tert-butyl (S) -2-amino-7-methyl-4- (methylthio) -5-oxo-5, 7-dihydro-6H-pyrrolo [3, 4-d] pyrimidine-6-carboxylate

A chiral purification was performed on tert-butyl 2-amino-7-methyl-4- (methylthio) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) -carboxylate (Intermediate I-12； 950 mg) via chiral SFC (Stationary phase: Chiralpak AD-H 5 μm 250 x 30 mm, Mobile phase: 80％CO₂, 20％iPrOH) to give the title compound (as the *Sisomer； 390 mg, 41％yield) . MS (ESI) : mass calcd. for C₁₃H₁₈N₄O₃S, 310.11； m/z found, 311.1 [M+H] ⁺. ¹H NMR (400 MHz, CDCl₃) δ 5.47 (s, 2H) , 4.75 (q, J ＝ 6.6 Hz, 1H) , 2.52 (s, 3H) , 1.61 -1.52 (m, 12H) .

Intermediate I-12-A2: tert-butyl (R) -2-amino-7-methyl-4- (methylthio) -5-oxo-5, 7-dihydro-6H-pyrrolo [3, 4-d] pyrimidine-6-carboxylate

A chiral purification was performed on tert-butyl 2-amino-7-methyl-4- (methylthio) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) -carboxylate (Intermediate I-12； 950 mg) via chiral SFC (Stationary phase: Chiralpak AD-H 5 μm 250 x 30 mm, Mobile phase: 80％CO₂, 20％iPrOH) to give the title compound (as the *R isomer； 416 mg, 44％yield) . MS (ESI) : mass calcd. for C₁₃H₁₈N₄O₃S, 310.11； m/z found, 311.1 [M+H] ⁺. ¹H NMR (400 MHz, CDCl₃) δ 5.47 (s, 2H) , 4.75 (q, J ＝ 6.6 Hz, 1H) , 2.52 (s, 3H) , 1.61 -1.52 (m, 12H) .

Intermediate II-1: (2S, 3S) -3-Methyl-1-picolinoylazetidine-2-carboxylic acid

Step1: (S) -Methyl 3-methyl-2- (picolinamido) butanoate

To a solution of L-valine methyl ester hydrochloride (16.7 g, 100 mmol) , picolinic acid (13.5 g, 11 mmol) , HOBt (14.9 g, 110 mmol) and DIEA (38.7 g, 300 mmol) in DCM (200 mL) was added EDCI (21.1 g, 110 mmol) . The resulting mixture was stirred at rt. overnight, and then washed with water. The organic layer was concentrated to dryness, the resulting residue was purified by flash column chromatography to give the title compound as an oil (20.5 g) . Yield: 87％. MS (ESI) : calcd. value for C₁₂H₁₆N₂O₃ is 236.12, m/z measured value is 237.1 (M+1) ⁺.

Step2: (2S, 3S) -Methyl 3-methyl-1-picolinoylazetidine-2-carboxylate

To a solution of (S) -methyl 3-methyl-2- (picolinamido) butanoate (1.5 g, 6.36 mmol) , acetic acid (763 mg, 12.72 mmol) , and iodosobenzene diacetate (5.12 g, 15.9 mmol) in toluene (15 mL) , was added diacetoxypalladium (71 mg, 0.32 mmol) . The resulting mixture was heated at 110℃ in a sealed tube overnight, and then cooled to rt., and concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as oil (850 mg) . Yield: 57％. MS (ESI) : calcd. value for C₁₂H₁₄N2O₃ is 234.10, m/z measured value is 235.1 (M+1) ⁺.

Step3: (2S, 3S) -3-Methyl-1-picolinoylazetidine-2-carboxylic acid

To a solution of (2S, 3S) -methyl 3-methyl-1-picolinoylazetidine-2-carboxylate (2.55 g, 10.9 mmol) in THF (13 mL) was added NaOH solution (1N, 13 mL) . The resulting mixture was stirred at rt. for 3 h, and then neutralized by 1N HCl (a.q. ) . The mixture was concentrated to dryness to afford the crude title product as a solid. ¹H NMR (400 MHz, CDCl₃) δ 8.55 (d, J ＝ 4.0 Hz, 0.5H) , 8.43 (d, J ＝ 4.0 Hz, 0.5H) , 8.16 (d, J ＝ 8.0 Hz, 0.5H) , 8.08 (d, J ＝ 7.6 Hz, 0.5H) , 7.90 (t, J ＝ 7.2 Hz, 0.5H) , 7.79 (t, J ＝ 7.2 Hz, 0.5H) , 7.47-7.44 (m, 0.5H) , 7.38-7.35 (m, 0.5H) , 4.76-4.68 (m, 1H) , 4.36-4.32 (m, 0.5H) , 4.25-4.20 (m, 0.5H) , 3.76-3.72 (m, 0.5H) , 3.12-3.02 (m, 1H) . MS (ESI) : calcd. value for C₁₁H₁₂N₂O₃ is 220.08, m/z measured value is 221.1 (M+1) ⁺.

The following intermediates were prepared according to the procedure of Intermedaite II-1 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:

Intermediate II-4: (2S, 3R) -1- (tert-butoxycarbonyl) -3-ethylazetidine-2-carboxylic acid

To a mixture of Intermediate II-2 (was prepared according the procedure of Intermediate II-1 from (2S, 3S) -methyl 2-amino-3-methylpentanoate hydrochloride) (17.7 g, 71 mmol) in MeOH (150 mL) and water (150 mL) , was added potassium hydroxide (8.0 g, 142 mmol) at 5℃. The resulting mixture was warmed to rt. and stirred for 2 h, and then neutralized by 4N HCl (aq. ) . The mixture was concentrated to remove MeOH, and added conc. HCl (aq., 50 mL) . The resulting mixture was refluxed overnight, and then concentrated to dryness. The residue was dissolved in the mixture of water (100 mL) and THF (100 mL) , and added sodium hydroxide (4.3 g, 107 mmol) . The mixture was stirred at rt. for 30 min, and then di-tert-butyl dicarbonate (18.6 g, 85 mmol) was added. The resulting mixture was stirred at rt. for 2 h. The reaction mixture was washed with ethyl acetate twice, the aqueous phase was neutralized by 1N HCl (aq. ) to PH＝2 carefully, and then extracted with EtOAc. The organic layers were combined, and then concentrated under vacuum to afford the title compound (9.1 g) as an oil, which was used directly in the next step without purification.

300 mg of crude (2S, 3R) -1- (tert-butoxycarbonyl) -3-ethylazetidine-2-carboxylic acid was purified by column chromatography to afford pure title compound as an oil (170 mg) . ¹HNMR: (400 MHz, CD₃OD) δ 4.63 (d, J ＝ 9.2 Hz, 1H) , 3.95 –3.88 (m, 1H) , 3.60 –3.51 (m, 3H) , 2.87 –2.69 (m, 1H) , 1.45 –1.36 (m, 10H) , 0.86 (t, J ＝ 7.2 Hz, 3H) . MS (ESI) : calcd value for C₁₁H₁₉NO₄ is 229.13, m/z measured value is 228.1 (M-1) ^-.

Intermediate II-5: (2S, 3S) -1- (tert-butoxycarbonyl) -3- (trifluoromethyl) azetidine-2-carboxylic acid

Step 1: (2-Bromopyridin-3-yl) methanol

NaBH₄ (12.2 g, 322 mmol) was added in portions to a 0 ℃ (ice/water) mixture consisting of 2-bromo-3-formylpyridine (50.0 g, 269 mmol) and dry MeOH (400 mL) . The resultant mixture was stirred for 2 hours with gradual warming to 25 ℃ before quenching with H₂O (500 mL) , and removing the MeOH under reduce pressure. The concentrate was extracted with ethyl acetate (500 mL x 3) , and the combined organic extracts dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness under reduced pressure to afford the title compound (48.0 g) . Yield: 90.2％. ¹H NMR (400 MHz, CDCl₃) : δ 8.27 (dd, J ＝ 1.6, 4.4 Hz, 1H) , 7.86 (d, J ＝ 7.6 Hz, 1H) , 7.31 (dd, J ＝ 5.0, 7.6 Hz, 1H) , 4.75 (d, J ＝ 5.6 Hz, 2H) , 2.66 (t, J ＝ 6.0 Hz, 1H) .

Step 2: 2-Bromo-3- ( ( (tert-butyldimethylsilyl) oxy) methyl) pyridine

TBSCl (46.2 g, 306 mmol) was added in portions to a 0 ℃ (ice/water) mixture consisting of (2-bromopyridin-3-yl) methanol (48.0 g, 255 mmol) , imidazole (52.1 g, 766 mmol) and dichloromethane (500 mL) . The resultant mixture was stirred for 16 hours with gradual warming to room-temperature before diluting with dichloromethane (500 mL) and washing with brine (300 mL x 3) . The organic phase was dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness under reduced pressure to give the crude product, which was purified by fast column chromatography to afford the title compound (64.3 g) . Yield: 79.2％. ¹H NMR (400 MHz, CDCl₃) : δ 8.26 (d, J ＝ 3.4 Hz, 1H) , 7.86 (d, J ＝ 7.4 Hz, 1H) , 7.31 (dd, J ＝ 4.6, 7.6 Hz, 1H) , 4.70 (s, 2H) , 0.98 (s, 9H) , 0.15 (s, 6H) .

Step 3: Benzyl 3- ( ( (tert-butyldimethylsilyl) oxy) methyl) picolinate

2-Bromo-3- ( ( (tert-butyldimethylsilyl) oxy) methyl) pyridine (64.3 g, 213 mmol) , benzyl alcohol (115 g, 1.06 mol) , triethylamine (108 g, 1.06 mol) , Pd (dppf) Cl₂·CH₂Cl₂ (17 g, 21 mmol) and DMF (400 mL) were added to a 1 L autoclave. The resultant mixture was stirred and heated at 60 ℃ under CO (1.4 bar) for 48 hours before pouring into H₂O (600 mL) and extracting with ethyl acetate (500 mL x 3) . The combined organic extracts were washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated to dryness under reduced pressure to give the crude product, which was purified by fast column chromatography to afford the title compound (35 g) . Yield: 53％. MS (ESI) : mass calcd. for C₂₀H₂₇NO₃Si 357.18_, m/z found 358.0 [M+H] ⁺.

Step 4: 3- ( ( (tert-Butyldimethylsilyl) oxy) methyl) picolinic acid

Benzyl 3- ( ( (tert-butyldimethylsilyl) oxy) methyl) picolinate (25.0 g, 69.9 mmol) , ethyl acetate (400 mL) and wet Pd/C (5.0 g, 10 wt％, 4.6 mmol) were added to a 1 L round-bottomed flask. The resultant mixture was stirred under H₂ (1 atm) at 25 ℃ for 18 hours. The suspension was filtered through a pad of

and the pad then washed with ethyl acetate (100 mL) . The filtrate was concentrated to dryness under reduced pressure to afford the title product (15.4 g, crude) , which was used for next step without further purification. MS (ESI) : mass calcd. for C₁₃H₂₁NO₃Si 267.13_, m/z found 267.9 [M+H] ⁺.

Step 5: (Z) -Methyl 4, 4, 4-trifluoro-2-formamido-3-methylbut-2-enoate

A solution consisting of methyl isocyanoacetate (50.0 g, 505 mmol) and dry THF (100 mL) was added drop-wise to a -60 ℃ (dry ice/acetone) mixture consisting of t-BuONa (53.3 g, 555 mmol) and dry THF (300 mL) . The resultant mixture was stirred at -60 ℃ for 30 minutes and then treated with a solution consisting of trifluoromethyl ketone (71.8 g, 641 mmol) and dry THF (190 mL) . The resulting mixture was stirred at -60 ℃ for 1 hour and stirred for another 1 hour with gradual warming to 25 ℃ before acidifying with 1 M HCl to pH ＝ 4-5. The THF was then removed under reduced pressure and the residue extracted with ethyl acetate (400 mL x 3) . The combined organic extracts were washed with brine (600 mL) , dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness under reduced pressure to give the crude product, which was purified by fast column chromatography to afford the title compound (61 g) . Yield: 54％. ¹H NMR (400 MHz, DMSO-d₆) : δ 10.21 (br.s., 1H) , 8.09 (d, J ＝ 0.8 Hz, 1H) , 3.72 (s, 3H) , 1.88 (s, 3H) .

Step 6: (2S, 3S) -Methyl 4, 4, 4-trifluoro-2-formamido-3-methylbutanoate

(Z) -Methyl 4, 4, 4-trifluoro-2-formamido-3-methylbut-2-enoate (14 g, 66 mmol) , MeOH (500 mL) and (-) -1, 2-bis ( (2S, 5S) -2, 5-dimethylphospholano) ethane (cyclooctadiene) -rhodium (I) tetrafluoroborate (959 mg, 1.72 mmol) were added to a 1 L hydrogenation bottle. The resultant mixture was stirred under H₂ (55 psi) at room-temperature for 72 hours. The suspension was filtered through a pad of

and the pad washed with ethyl acetate (100 mL) . The filtrate was concentrated to dryness under reduced pressure to give the crude product, which was purified by fast column chromatography to afford the title compound (13 g) . Yield: 87％. ¹H NMR (400 MHz, CDCl₃) : δ 8.25 (s, 1H) , 6.42 (s, 1H) , 5.09 (dd, J ＝ 3.4, 9.0 Hz, 1H) , 3.81 (s, 3H) , 2.94-2.82 (m, 1H) , 1.22 (d, J ＝ 7.2 Hz, 3H) .

Step 7: (2S, 3S) -Methyl 2-amino-4, 4, 4-trifluoro-3-methylbutanoate hydrochloride

(2S, 3S) -Methyl 4, 4, 4-trifluoro-2-formamido-3-methylbutanoate (13 g, 61 mmol) , MeOH (260 mL) and 12.1 M HCl (6.6 mL, 79 mmol) were added to a 1 L round-bottomed flask. The resultant mixture was stirred at 45 ℃ to 50 ℃ for 5 hours before concentrating to dryness under reduced pressure and triturating with MTBE (130 mL) and methanol (1.3 mL) . The suspension was isolated via filtration and the filter cake washed with MTBE (200 mL) before drying under reduced pressure to afford the title compound (11.6 g) . Yield: 81.4％. ¹H NMR (400 MHz, CD₃OD) : δ 4.50 (d, J ＝ 3.2 Hz, 1H) , 3.90 (s, 3H) , 3.29-3.21 (m, 1H) , 1.30 (d, J ＝ 7.4 Hz, 3H) .

Step 8: (2S, 3S) -Methyl 2- (3- ( ( (tert-butyldimethylsilyl) oxy) methyl) picolinamido) -4, 4, 4-trifluoro-3-methylbutanoate

HATU (25 g, 66 mmol) was added to a solution consisting of (2S, 3S) -methyl 2-amino-4, 4, 4-trifluoro-3-methylbutanoate hydrochloride (9.8 g, 44 mmol) , 3- ( ( (tert-butyldimethylsilyl) oxy) methyl) picolinic acid (14 g, 53 mmol) , DIPEA (28.6 g, 222 mmol) , and dichloromethane (200 mL) . The resultant mixture was stirred at room-temperature for 16 hours before concentrating to dryness under reduced pressure to give the crude product, which was purified by FCC to afford the title compound (14.2 g, 73.5％) . MS (ESI) : mass calcd. for C₁₉H₂₉F₃N₂O₄Si 434.18, m/z found 435.1 [M+H] ⁺.

Step 9: (2S, 3S) -Methyl 1- (3- ( ( (tert-butyldimethylsilyl) oxy) methyl) picolinoyl) -3- (trifluoro methyl) azetidine-2-carboxylate

(2S, 3S) -Methyl 2- (3- ( ( (tert-butyldimethylsilyl) oxy) methyl) picolinamido) -4, 4, 4-trifluoro -3-methylbutanoate (14 g, 33 mmol) and PhI (OAc) ₂ (26.2 g, 81.5 mmol) were added to a 250 mL sealed tube and the resultant mixture dissolved in toluene (150 mL) . The mixture was sparged with N₂ for 5 minutes and then treated with Pd (OAc) ₂ (366 mg, 1.63 mmol) . The mixture was sparged with N₂ for another 5 minutes and then stirred and heated at 110 ℃ for 16 hours before cooling to room-temperature, pouring into H₂O (1 L) and extracting with dichloromethane (200 mL x 3) . The combined organic extracts were dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness under reduced pressure to give the crude product, which was purified by fast column chromatography to afford the title compound (10.4 g) . Yield: 72.4％. MS (ESI) : mass calcd. for C₁₉H₂₇F₃N₂O₄Si 432.17, m/z found 433.0 [M+H] ⁺.

Step 10: (2S, 3S) -1-tert-Butyl 2-methyl 3- (trifluoromethyl) azetidine-1, 2-dicarboxylate

1 M HCl (57.8 mL, 57.8 mmol) was added to a solution consisting of (2S, 3S) -methyl 1- (3- ( ( (tert-butyldimethylsilyl) oxy) methyl) picolinoyl) -3- (trifluoromethyl) azetidine-2-carboxylate (5.0 g, 12 mmol) and 1, 4-dioxane (240 mL) . The resultant mixture was stirred at room-temperature for 16 hours before treating with sodium carbonate (12.3 g, 116 mmol) and Boc₂O (5.0 g, 23 mmol) . The resulting mixture was stirred at room-temperature for 16 hours before diluting with H₂O (200 mL) and extracting with ethyl acetate (200 mL x 3) . The combined organic extracts were dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness under reduced pressure to give the crude product, which was purified by fast column chromatography to afford the title compound (2.5 g) . Yield: 73％. ¹H NMR (400 MHz, CDCl₃) : δ 4.66 (d, J ＝ 5.2 Hz, 1H) , 4.18 (t, J ＝ 8.8 Hz, 1H) , 3.95 (dd, J ＝ 5.4, 8.8 Hz, 1H) , 3.83 (s, 3H) , 3.25 -3.13 (m, 1H) , 1.44 (s, 9H) .

Step 11: (2S, 3S) -1- (tert-Butoxycarbonyl) -3- (trifluoromethyl) azetidine-2-carboxylic acid

1 M NaOH (17.7 mL, 17.7 mmol) was added drop-wise to a 0 ℃ (ice/water) solution consisting of (2S, 3S) -1-tert-butyl 2-methyl 3- (trifluoromethyl) azetidine-1, 2-dicarboxylate (2.5 g, 8.8 mmol) and THF (70 mL) . The resultant mixture was stirred for 2 hours with gradual warming to room-temperature before acidifying with 0.5 M HCl to pH ＝ 4-5 and extracting with ethyl acetate (120 mL x 3) . The combined organic extracts were dried over anhydrous Na₂SO₄, filtered, and concentrated to dryness under reduced pressure to afford the title compound (1.9 g) . Yield: 79％. ¹H NMR (400 MHz, DMSO-d₆) : δ 4.44 (d, J ＝ 5.0 Hz, 1H) , 4.09 (t, J ＝ 8.8 Hz, 1H) , 3.84-3.74 (m, 1H) , 3.69-3.55 (m, 1H) , 1.37 (s, 9H) .

Intermediate II-6: (2S, 3R) -1-benzyl-3-methylazetidine-2-carboxylic acid

Step 1: (S) -1- (benzylamino) propan-2-ol

In a flask was placed with (S) -1-aminopropan-2-ol (750mg, 10mmol) and benzaldehyde (1060mg, 10mmol) in EtOH (30mL) , then added NaBH3CN (1260mg, 20mmol) , stirred at rt. for 1h. The mixture was purified by fast column chromatography to afford the title compound (1.65 g) as yellow oil. Yield: 100％. MS (ESI) : calcd. value for C₁₀H₁₅NO is 165.24, m/z measured value is 166.2 (M+1) ⁺.

Step 2: tert-butyl (S) -N-benzyl-N- (2-hydroxypropyl) glycinate

To a mixture of (S) -1- (benzylamino) propan-2-ol (1.4 g, 8.5 mmol) , K₂CO₃ (2.4 g, 17 mmol) and NaI (2.6 g, 17mmol) in dry DMF (50 mL) , tert-butyl 2-bromoacetate (3.3 g, 17 mmol) was added. The resulting mixture was stirred at r.t. overnight. The mixture was diluted with DCM and the solid was filtered. The filtrate was concentrated under reduce pressure. The residue was purified by fast column chromatography to afford the title compound as a light yellow oil (1.5g) . Yield: 63％. MS (ESI) : mass calcd. for C₁₆H₂₅NO₃ 279.38, m/z found 280.2 [M+H] ⁺.

Step 3: tert-butyl (S) -N-benzyl-N- (2-chloropropyl) glycinate

To a solution of tert-butyl (S) -N-benzyl-N- (2-hydroxypropyl) glycinate (1.4 g, 8.5 mmol) in DCM (50 mL) , SOCl₂ (5 mL) was added. The reaction was refluxed 4h. The solution was concentrated under reduce pressure. The residue was dissolved in DCM, washed with sat. NaHCO₃. The aqueous layer was extracted with DCM. The organic phases were combined, and then concentrated under vacuum. The residue was purified by fast column chromatography to afford the title compound as a colorless oil (950mg) . Yield: 38％. MS (ESI) : mass calcd. for C₁₆H₂₄ClNO₂ 297.82_, m/z found 298.1 [M+H] ⁺.

Step 4: tert-butyl (2S, 3R) -1-benzyl-3-methylazetidine-2-carboxylate

To a solution of tert-butyl (S) -N-benzyl-N- (2-chloropropyl) glycinate (950 mg, 3.2 mmol) in dry THF (30 mL) and HMPA (7 mL) , LiHDMS (6.4 mL, 1N in THF, 6.4 mmol) was added dropwise at -70℃. The resulting solution was allowed to warm slowly to 0℃ and stirred at that temperature for 1h. The reaction was quenched with sat. NH₄Cl. The aqueous phase was extracted with EA, the organic phases were combined, and then concentrated under vacuum. The residue was purified fast comlumn chromatography to afford the title compound as yellow oil (523mg) . Yield: 63％. MS (ESI) : mass calcd. for C₁₆H₂₃NO₂ 261.17, m/z found 262.2 [M+H] ⁺. ¹H NMR (400 MHz, CDCl₃) : δ 7.32 –7.18 (m, 5H) , 3.74 (d, J ＝ 12.4 Hz, 1H) , 3.67 (d, J ＝ 8.4 Hz, 1H) , 3.57 (d, J ＝ 12.4 Hz, 1H) , 3.03 –2.99 (m, 1H) , 2.97 –2.94 (m, 1H) , 2.68 –2.57 (m, 1H) , 1, 38 (s, 9H) , 1.22 (d, J ＝ 7.2 Hz, 3H) .

Step 5: (2S, 3R) -1-benzyl-3-methylazetidine-2-carboxylic acid

To a solution of tert-butyl (2S, 3R) -1-benzyl-3-methylazetidine-2-carboxylate (523 mg, 2 mmol) in DCM (30 mL) , TFA was added. The resulting solution was stirred at r.t. overnight. Solvent was evaporated and the residue using for next step without purification. MS (ESI) : mass calcd. for C₁₂H₁₅NO₂ 205.26, m/z found 206.1 [M+H] ⁺.

The following intermediates are commercial available:

Intermediate III-A1: (S) -tert-butyl 2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidine-1-carboxylate

Step 1: methyl 1-amino-3-chloro-1H-pyrrole-2-carboxylate

To a mixture of NaH (87 g , 2.19 mol) in DMF (500 mL) was added a solution of methyl 3-chloro-1H-pyrrole-2-carboxylate (250 g, 1.56 mol) in DMF (1200 mL) dropwise at 0℃. The mixture was stirred for 1.5 h at 0℃, then a solution of O- (2, 4-dinitrophenyl) hydroxylamine (466 g, 2.34 mol) in DMF (1200 mL) was added dropwise at 0℃. The resulting mixture was warmed to r.t. slowly, and stirred for 3 h at r.t. The mixture was poured into ice-water, and extracted with EtOAc. The organic layers were concentrated and purified by column chromatography to afford the title compound as a solid (274 g) . MS (ESI) : calcd. value for C₆H₇ClN₂O₂ is 174.02, m/z measured value is 174.9 (M+1) ⁺.

Step 2: 1-amino-3-chloro-1H-pyrrole-2-carboxamide

The solution of methyl 1-amino-3-chloro-1H-pyrrole-2-carboxylate (180 g, 1.03 mol) dissolved in 1.5 L of 7N NH₃/MeOH (10.31 mol) was stirred in a sealed flask at 130℃ for 48 h. After cooling to rt., the mixture was concentrated to dryness and crystallized in MeOH to afford the title compound as a solid (92 g) . Yield: 56％. MS (ESI) : calcd. value for C₅H₆ClN₃O is 159.02, m/z measured value is 160.0 (M+1) ⁺.

Step 3: (S) -tert-butyl 2- ( (2-carbamoyl-3-chloro-1H-pyrrol-1-yl) carbamoyl) azetidine-1-carboxylate

To a mixture of 1-amino-3-chloro-1H-pyrrole-2-carboxamide (187 g, 1.18 mol) in DCM (5 L) were added (S) -1- (tert-butoxycarbonyl) azetidine-2-carboxylic acid (260 g, 1.29 mol) and EDCI. HCl (248 g, 1.29 mol) . The resulting mixture was stirred for 3 days at rt. The mixture was washed with water and brine. The organic layers were dried over Na₂SO₄, and then concentrated to dryness to afford the title compound as oil. MS (ESI) : calcd. value for C₁₄H₁₉ClN₄O₄ is 342.11, m/z measured value is 243.1 (M-Boc+1) ⁺.

Step 4: (S) -tert-butyl 2- (5-chloro-4-oxo-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidine-1-carboxylate

To a mixture of (S) -tert-butyl 2- ( (2-carbamoyl-3-chloro-1H-pyrrol-1-yl) carbamoyl) azetidine-1-carboxylate (100 g, 294 mmol) in EtOH (1.47 L) was added 1.47 L of aq. KOH (1.47 mol, 1 N) , the resulting mixture was stirred at 90℃ overnight. The mixture was adjusted to pH＝2～3 with aq. HCl (1 N) at 0℃, the resulting precipitate was filtered and dried to afford the title compound (75 g) as a solid. Yield: 79％. MS (ESI) : calcd. value for C₁₄H₁₇ClN₄O₃ is 324.10, m/z measured value is 324.7 (M+1) ⁺.

Step 5: (S) -tert-butyl 2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidine-1-carboxylate

To a solution of (S) -tert-butyl 2- (5-chloro-4-oxo-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidine-1-carboxylate (50 g, 154 mmol) in DCM (500 mL) were added phenylboronic acid (56.3 g, 462 mmol) , Py (50 mL) , Cu (OAc) ₂ (84 g, 462 mmol) and some

molecular sieve, then purged by O₂, the resulting mixture was stirred at 30℃ for 48 h under O₂. The mixture was diluted with DCM (500 mL) , and filtered, the resulting filtrate was concentrated to dryness and purified by column chromatography to afford the title compound as a solid (30 g) . Yield: 49％. MS (ESI) : calcd. value for C₂₀H₂₁ClN₄O₃ is 400.13, m/z measured value is 400.9 (M+1) ⁺.

Intermedaite III-A1 can also be prepared according to the procedure of Intermedaite III-B1 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art.

Intermediate III-B1: (2S, 3S) -tert-butyl 2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidine-1-carboxylate

Step 1: (2S, 3S) -tert-butyl 3-methyl-2- (phenylcarbamoyl) azetidine-1-carboxylate

To a solution of Intermediate II-1 (16.0 g, 74.3 mmol) in DCM (300 mL) were added aniline (6.8 mL, 74.3 mmol) and EDCI. HCl (15.7 g, 81.7 mmol) . The resulting mixture was stirred at rt. overnight, and then washed with water. The organic layer was concentrated to dryness and purified by column chromatography to afford the title compound as a solid (20 g) . Yield: 92％. MS (ESI) : calcd. value for C₁₆H₂₂N₂O₃ is 290.16, m/z measured value is 313.0 (M+23) ⁺.

Step 2: (2S, 3S) -tert-butyl 2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidine-1-carboxylate

To the solution of Ph₃P (29.8 g, 113.6 mmol) in DCM (150 mL) was added I₂ (26.2 g, 103.3 mmol) in portions at rt. under N₂. The resulting mixture was stirred at rt. for 0.5 h under N₂, and then, Et₃N (43.2 mL, 310.0 mmol) was added. The mixture was stirred at rt. for 0.5 h under N₂, and then a solution of (2S, 3S) -tert-butyl 3-methyl-2- (phenylcarbamoyl) azetidine-1-carboxylate (15.0 g, 51.7 mmol) in DCM (75 mL) was added dropwise. The mixture was stirred at rt. for 6.5 h under N_2, and then a solution of methyl 1-amino-3-chloro-1H-pyrrole-2-carboxylate (9 g, 51.7 mmol) in DCM (75 mL) was added dropwise. The mixture was stirred at rt. for 80 h under N₂, and then concentrated to dryness and purified by column chromatography to afford the title compound as a solid (15.0 g) . Yield: 70％. MS (ESI) : calcd. value for C₂₁H₂₃ClN₄O₃ is 414.15, m/z measured value is 415.1 (M+1) ⁺.

Intermediate III-B1 can also be prepared according to the procedure of intermedaite III-A1 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art.

Intermediate III-I1: tert-butyl (2S, 3S) -2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-ethylazetidine-1-carboxylate

Step 1: methyl (S) -2- (benzylamino) butanoate

To a solution of (S) -2- ( (tert-butoxycarbonyl) amino) butanoic acid (60.9 g, 300 mmol) in MeOH (600 mL) , Con. H₂SO₄ (25 mL) was added dropwise. The resulting solution was refluxed 2h. The reaction was cooled to r.t., and then diluted with water, neutralized by Na₂CO₃. The mixture was extracted with DCM. The organic phase was concentrated under reduce pressure to give a yellow oil. This product was dissolved in EtOH (600 mL) , and then Benzaldehyde (35.1 g, 330 mmol) and AcOH (15mL) were added. At last, NaBH₃CN (22.5 g, 120 mmol) was added. The resulting mixture was stirred at r.t. for 2h. The reaction was quenched with HCl (3M) and acidified to pH＝1. EtOH was evaporated and the aqueous solution was washed with EA twice, and then neutralized again by Na₂CO₃ to about PH＝12. The mixture was extracted DCM, the organic phase was concentrated under vacuum to afford the title compound as a colorless oil (24.3g) . Yield: 39％. MS (ESI) : calcd. value for C₁₂H₁₇NO₂ is 207.27, m/z measured value is 208.1 (M+1) ⁺.

Step 2: (S) -2- (benzylamino) butan-1-ol

To a mixture of LiAlH₄ (8.7 g, 231.6 mmol) in dry THF (450 mL) , methyl (S) -2- (benzylamino) butanoate (24 g, 115.8 mmol) was added slowly at 5℃. The resulting mixture was stirred at the same temperature for 2h. Ice was added slowly into the reaction. The resulting mixture was stirred at r.t. for 10min. The precipitate was filtered and the filtrate was extracted with EA. The organic layers were combined, and then concentrated under reduce pressure. The residue was purified by column chromatography to afford the title compound as a light yellow oil (14.4g) . Yield: 70％. MS (ESI) : calcd. value for C₁₁H₁₇NO is 179.26, m/z measured value is 180.1 (M+1) ⁺.

Step 3: (S) -2- (benzyl (1-hydroxybutan-2-yl) amino) acetonitrile

To a mixture of (S) -2- (benzylamino) butan-1-ol (4.8g, 26.8mmol) and Na₂CO₃ (8.5g, 80.4mmol) in MeCN (200 mL) , 2-bromoacetonitrile (4.8g, 40.2mmol) was added. The resulting mixture was stirred at 35℃ overnight. The mixture was diluted with DCM and the solid was filtered. The filtrate was concentrated under reduce pressure. The residue was purified by column chromatography to afford the title compound as a light yellow oil (5.0g) . Yield: 86％. MS (ESI) : calcd. value for C₁₃H₁₈N₂O is 218.14, m/z measured value is 219.1 (M+1) ⁺.

Step 4: (R) -2- (benzyl (2-chlorobutyl) amino) acetonitrile

To a solution of (S) -2- (benzyl (1-hydroxybutan-2-yl) amino) acetonitrile (4.0 g, 15.9mmol) in DCM (100 mL) , SOCl₂ (15 mL) was added. The resulting solution was refluxed overnight. The reaction was cooled to r.t., concentrated under vacuum. The residue was dissolved in DCM, washed with sat. NaHCO₃. The aqueous layer was extracted with DCM. The organic phases were combined, and then concentrated under vacuum. The residue was purified by column chromatography to give a light yellow oil (2.4g) . The product was dissolved in DMF (50mL) , the resulting solution was stirred at 65℃ for 3 days. DMF was evaporated under vacuum to afford the title compound as a yellow oil (4.0 g) which was used directly in the next step. Yield: 100％. MS (ESI) : calcd. value for C₁₃H₁₇N₂Cl is 236.11.

Step 5: (3S) -1-benzyl-3-ethylazetidine-2-carbonitrile

To a solution of (R) -2- (benzyl (2-chlorobutyl) amino) acetonitrile (6.4 g, 27.1 mmol) in dry THF (350 mL) , LiHMDS (55 mL, 1N in THF) was added dropwise at -60℃. The resulting solution was allowed to warm slowly to 0℃ and stirred for 1h at that temperature. The reaction was quenched with MeOH, concentrated under vacuum. The residue was purified by column chromatography to afford the title as a yellow oil (3.2g) . Yield: 60％, MS (ESI) : calcd. value for C₁₃H₁₆N₂ is 200.13, m/z measured value is 201.2 (M+1) ⁺.

Step 6: (3S) -1-benzyl-3-ethylazetidine-2-carboxylic acid

To a solution of (3S) -1-benzyl-3-ethylazetidine-2-carbonitrile (1.2 g, 6.0 mmol) in EtOH (50 mL) and water (50 mL) , NaOH (4.0 g, 100mmol) was added. The resulting solution was refluxed 24h. LCMS indicated that the reaction was complete. The solution was cooled to r.t., and then acidified with con. HCl to pH＝1. The resulting mixture was concentrated under vacuum. The residue was purified by column chromatography to afford the title compound as a white solid (850mg) . Yield: 57％. MS (ESI) : calcd. value for C₁₃H₁₇NO₂ is 219.28, m/z measured value is 220.1 (M+1) ⁺.

Step 7: (3S) -1-benzyl-3-ethyl-N-phenylazetidine-2-carboxamide

To a solution of (3S) -1-benzyl-3-ethylazetidine-2-carboxylic acid (850 mg, 3.9 mmol) , aniline (548 mg, 5.9 mmol) , HOBt (1.1 g, 7.8 mmol) and DIEA (2.5 g, 19 mmol) in DCM (50 mL) , EDCI·HCl (1.5 g, 7.8 mmol) was added. The resulting mixture was stirred at r.t. overnight. LC-MS indicated that the reaction was complete. The reaction solution was concentrated under reduce pressure. The residue was purified by column chromatography to afford the title compound as a yellow solid (750mg) . Yield: 65％. MS (ESI) : calcd. value for C₁₉H₂₂N₂O is 294.40, m/z measured value is 295.2 (M+1) ⁺.

Step 8: tert-butyl (2S, 3S) -3-ethyl-2- (phenylcarbamoyl) azetidine-1-carboxylate

To a solution of (3S) -1-benzyl-3-ethyl-N-phenylazetidine-2-carboxamide (750 mg, 2.5 mmol) , (Boc) ₂O (829 mg, 3.8 mmol) in EtOH (50 mL) , Pd (OH) ₂ (351 mg, 2.5 mmol) was added. The resulting mixture was stirred under hydrogen ballon pressure for 4h. After the completion of the reaction, the reaction mixture was filtered through Celite and the filtrate was concentrated and dried under vacuum. The crude residue was purified by column chromatography to afford the title compound (490 mg) and tert-butyl (2R, 3S) -3-ethyl-2- (phenylcarbamoyl) azetidine-1-carboxylate (50 mg) as a white solid. Yield: 64％. MS (ESI) : calcd. value for C₁₇H₂₄N₂O₃ is 304.39, m/z measured value is 205.1 (M-99) ⁺.

Step 9: tert-butyl (2S, 3S) -2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-ethylazetidine-1-carboxylate

To a solution of PPh₃ (1.1 g, 4.3 mmol) in DCM (30 mL) , I₂ (990 mg, 3.9 mmol) was added portion wise under N₂ atmosphere. The resulting solution was stirred at r.t. for 30min, then Et₃N (1.5g, 11.7mmol) was added and stirred for another 30min. To the mixture above was added tert-butyl (2S, 3S) -3-ethyl-2- (phenylcarbamoyl) azetidine-1-carboxylate (400 mg, 1.3 mmol) , the mixture was stirred for 6h, ethyl 1-amino-3-chloro-1H-pyrrole-2-carboxylate (282 mg, 1.5 mmol) was added and the result mixture was stirred for 2 days at rt.. The reaction mixture was concentrated under vacuum. The residue was purified by column chromatography to afford a yellow oil. This crude product was dissolved in DCM, Et₃N (1.5g, 11.7mmol) was added. The resulting mixture was refluxed 2h. LC-MS indicated that the reaction was complete. The reaction solution was concentrated under reduce pressure. The residue was purified by column chromatography to afford the title compound as a light yellow solid (490mg) . Yield: 88％. MS (ESI) : calcd. value for C₂₂H₂₅ClN₄O₃ is 428.92, m/z measured value is 429.2 (M+1) ⁺.

The following intermediates were prepared according to the procedures of intermedaite III-A1 or intermedaite III-B1 or intermidiate III-I1 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:

Intermediate III-K1: (S) -tert-butyl 2- (5-iodo-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidine-1-carboxylate

To a solution of intermediate III-E1 (300 mg, 0.67 mmol) in 1, 4-dioxane (8 mL) were added NaI (404 mg, 2.70 mmol) , trans-1, 2-bis (methylamino) cylcohexane (96 mg, 0.67 mmol) and CuI (64 mg, 0.337 mmol) , the resulting mixture was stirred at reflux for 3 days under N₂. The mixture was allowed to cool down to rt. and filtered through celite, and washed with ethyl acetate. The combined filtrate was concentrated to dryness and purified by flash column chromatography to afford the title compound as a solid (200 mg) . Yield: 60％. MS (ESI) : calcd. value for C₂₀H₂₁IN₄O₃ is 492.07, m/z measured value is 492.9 (M+1) ⁺.

The following intermediate was prepared according to the procedure of intermediate III-K1 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:

Intermediate III-L1: (S) -tert-butyl 2- (4-oxo-3-phenyl-5- (trifluoromethyl) -3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidine-1-carboxylate

To a mixture of Intermediate III-K1 (200 mg, 0.4 mmol) and CuI (94 mg, 0.49 mmol) were added DMF (5 mL) , HMPA (0.35 mL) and methyl 2, 2-difluoro-2- (fluorosulfonyl) acetate (0.25 mL, 2 mmol) under N₂ atmosphere. The resulting mixture was stirred under N₂ at 80℃ for 24 h. After cooling to rt., the mixture was poured into abundant ice-water and extracted with ethyl acetate. The organic mixture was washed with water and brine, and concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (165 mg) . Yield: 94％. MS (ESI) : calcd. value for C₂₁H₂₁F₃N₄O₃ is 434.16, m/z measured value is 456.9 (M+23) ⁺.

The following intermediates were prepared according to the procedure of intermediate III-L1 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:

Intermediate III-M1: (S) -tert-butyl 2- (5-cyano-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidine-1-carboxylate

To a solution of Intermediate III-E1 (178 mg, 0.4 mmol) in DMF (8 mL) was added Zn(CN) ₂ (234 mg, 2 mmol) and Pd (PPh₃) ₄ (184 mg, 0.16 mmol) under N₂. The mixture was stirred under microwave condition at 140℃ for 4 h. The resulting mixture was concentrated to dryness and purified by flash column chromatography to afford the title compound as a solid (140 mg) . Yield: 90％. MS (ESI) : calcd. value for C₂₁H₂₁N₅O₃ is 391.42, m/z measured value is 336.1 (M-56+1) ⁺.

The following intermediates were prepared according to the procedure of intermediate III-M1 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:

Intermediate III-N1: (R) -N- (1- (3-chloro-4-oxo-5-phenyl-4, 5-dihydropyrazolo [1, 5-a] pyrazin-6-yl) ethyl) -2-methylpropane-2-sulfinamide

Step 1: Methyl 4-chloro-1- (2-oxopropyl) -1H-pyrazole-5-carboxylate

To a solution of methyl 4-chloro-1H-pyrazole-5-carboxylate (4.8 g, 30 mmol) , 1-hydroxypropan-2-one (2.22 g, 30 mmol) and PPh₃ (7.86 g, 30 mmol) dissolved in THF (50 mL) , was added diisopropyl azodicarboxylate (6.06 g, 30 mmol in 10 mL THF) dropwise at 0℃ under N₂. The mixture was stirred at rt. for 3 h, and concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (1.2 g) . Yield: 18％. MS (ESI) : calcd. value for C₈H₉ClN₂O₃ is 216.03, m/z measured value is 217.0 (M+1) ⁺.

Step 2: 3-Chloro-6-methylpyrazolo [1, 5-a] pyrazin-4 (5H) -one

Methyl 4-chloro-1- (2-oxopropyl) -1H-pyrazole-5-carboxylate (1.2 g, 5.5 mmol) and ammonium acetate (4.28 g, 55 mmol) were dissolved in AcOH (20 mL) , the resulting mixture was stirred at 120℃ overnight, and then concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (1.0 g) . Yield: 98.4％. MS (ESI) : calcd. value for C₇H₆ClN₃O is 183.02, m/z measured value is 184.2 (M+1) ⁺.

Step 3: 3-Chloro-6-methyl-5-phenylpyrazolo [1, 5-a] pyrazin-4 (5H) -one

To a solution of 3-chloro-6-methylpyrazolo [1, 5-a] pyrazin-4 (5H) -one (1.0 g, 5.46 mmol) , phenylboronic acid (1.33 g, 10.92 mmol) , Cupric Acetate Anhydrous (1.98 g, 10.92 mmol) and pyridine (1.73g, 21.86mmol) dissolved in DCM (15 mL) , was added

MS (18 g) . The resulting mixture was stirred at 30℃ under O₂ overnight, and then concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (1.1 g) . Yield: 77.7％. MS (ESI) : calcd. value for C₁₃H₁₀ClN₃O is 259.05, m/z measured value is 260.0 (M+1) ⁺.

Step 4: 3-chloro-4-oxo-5-phenyl-4, 5-dihydropyrazolo [1, 5-a] pyrazine-6-carbaldehyde

3-chloro-6-methyl-5-phenylpyrazolo [1, 5-a] pyrazin-4 (5H) -one (259 mg, 1 mmol) and SeO₂ (222 mg, 2 mmol) were dissolved in dioxane (20 mL) , the mixture was stirred at reflux overnight. The resulting precipitate was filtered, and the filtrate was concentrated to dryness to afford the title compound as a solid (273 mg) , which was used for next step without purification. MS (ESI) : calcd. value for C₁₃H₈ClN₃O₂ is 273.03, m/z measured value is 274.1 (M+1) ⁺.

Step 5: (R) -N- ( (3-chloro-4-oxo-5-phenyl-4, 5-dihydropyrazolo [1, 5-a] pyrazin-6-yl) methylene) -2-methylpropane-2-sulfinamide

3-chloro-4-oxo-5-phenyl-4, 5-dihydropyrazolo [1, 5-a] pyrazine-6-carbaldehyde (273 mg, 1 mmol) , (R) -2-methylpropane-2-sulfinamide (121 mg, 1 mmol) and tetraethoxytitanium (456 mg, 2 mmol) were dissolved in THF (30 mL) , the resulting mixture was stirred at 80℃ under N₂ overnight. The mixture was concentrated and purified by flash column chromatography to afford the title compound as a solid (350 mg) . Yield: 93％. MS (ESI) : calcd. value for C₁₇H₁₇ClN₄O₂S is 376.08, m/z measured value is 376.9 (M+1) ⁺.

Step 6: (R) -N- (1- (3-chloro-4-oxo-5-phenyl-4, 5-dihydropyrazolo [1, 5-a] pyrazin-6-yl) ethyl) -2-methylpropane-2-sulfinamide

To a solution of (R) -N- ( (3-chloro-4-oxo-5-phenyl-4, 5-dihydropyrazolo [1, 5-a] pyrazin-6-yl) methylene) -2-methylpropane-2-sulfinamide (350 mg, 0.93 mmol) dissolved in THF (25 mL) was added methylmagnesium bromide (3 M in ether, 0.34 mL, 1.02 mmol) dropwise at 0℃ under N₂. The resulting mixture was stirred at rt. for 2 h, and then concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (300 mg) . Yield: 82.4％. MS (ESI) : calcd. value for C₁₈H₂₁ClN₄O₂S is 392.11, m/z measured value is 393.1 (M+1) ⁺.

Intermediate III-O1: (R) -N- (1- (3-chloro-5-oxo-6-phenyl-5, 6-dihydroimidazo [1, 2-c] pyrimidin-7-yl) ethyl) -2-methylpropane-2-sulfinamide

Step 1: 5-acetyl-4-hydroxy-2H-1, 3-thiazine-2, 6 (3H) -dione

The mixture of malonic acid (20.8 g, 200 mmol) , KSCN (20.0 g, 206 mmol) , Ac₂O (20.0 mL) and AcOH (80 mL) was stirred at r.t. overnight. Then H₂O (100 mL) was added and extracted with DCM: MeOH＝9: 1, the organic layer was dried and concentrated to afford the title compound as a yellow solid which was used in the next step without further purification (2.0 g) . Yield: 53％.

Step 2: 6-methyl-1-phenylpyrimidine-2, 4 (1H, 3H) -dione

To a solution of 5-acetyl-4-hydroxy-2H-1, 3-thiazine-2, 6 (3H) -dione (20 g, 106 mmol) in DMF (15 mL) was added aniline (9.2 mL) at r.t., the reaction was stirred at reflux until 5-acetyl-4-hydroxy-2H-1, 3-thiazine-2, 6 (3H) -dione disappeared by TLC. The mixture was concentrated, the residue was washed with EtOH, and filtered to afford the title compound as a yellow solid (880 mg) . Yield: 40.7％. MS (ESI) : calcd. value for C₁₁H₁₀N₂O₂ is 202.21, m/z measured value is 203.1 (M+1) ⁺.

Step 3: 4-amino-6-methyl-1-phenylpyrimidin-2 (1H) -one

The solution of 6-methyl-1-phenylpyrimidine-2, 4 (1H, 3H) -dione (7.29 g, 36 mmol) in CH₃CN (120 mL) was purged by NH₃ for 5 min, then BOP (20.7 g, 46.8 mmol) and DBU (8.21 g, 54 mmol) were added, the reaction was stirred overnight. The mixture was filtered to afford the title compound as a white solid (7.24 g) . Yield: 100％. MS (ESI) : calcd. value for C₁₁H₁₁N₃O is 201.23, m/z measured value is 201.7 (M+1) ⁺.

Step 4: 7-methyl-6-phenylimidazo [1, 2-c] pyrimidin-5 (6H) -one

To a solution of 4-amino-6-methyl-1-phenylpyrimidin-2 (1H) -one (7.24 g, 36 mmol) in EtOH (100 mL) was added 40％2-chloroacetaldehyde in water (17.8 mL, 108 mmol) , the reaction was stirred at 100 ℃ overnight. The mixture was concentrated and purified by flash column chromatography to afford the title compound as a white solid (6.2 g) . Yield: 77％. MS (ESI) : calcd. value for C₁₃H₁₁N₃O is 225.25, m/z measured value is 225.9 (M+1) ⁺.

Step 5: 3-chloro-7-methyl-6-phenylimidazo [1, 2-c] pyrimidin-5 (6H) -one

7-methyl-6-phenylimidazo [1, 2-c] pyrimidin-5 (6H) -one (2.25 g, 10 mmol) and NCS (700 mg, 5.26 mmol) were dissolved in DMF (10 mL) , the reaction was stirred at r.t. for 3 h. The mixture was poured into H₂O (100 mL) , and extracted with EtOAc, the organic layers were washed with brine, dried over anhydrous Na₂SO₄ and concentrated. The resulting residue was washed with MeOH to afford the title compound as a white solid (600 mg) . Yield: 23％. MS (ESI) : calcd. value for C₁₃H₁₀ClN₃O is 259.69, m/z measured value is 260.1 (M+1) ⁺.

Step 6: 3-chloro-5-oxo-6-phenyl-5, 6-dihydroimidazo [1, 2-c] pyrimidine-7-carbaldehyde

3-chloro-7-methyl-6-phenylimidazo [1, 2-c] pyrimidin-5 (6H) -one (600 mg, 2.3 mmol) and SeO₂ (257 mg, 2.3 mmol) were dissolved in dioxane (20 mL) , the reaction was stirred at reflux overnight, then concentrated and purified by flash column chromatography to afford the title compound as a white solid (250 mg) . Yield: 39％. MS (ESI) : calcd. value for C₁₃H₈ClN₃O₂ is 273.68, m/z measured value is 274.1 (M+1) ⁺.

Intermediate III-O1 was prepared according to the procedure of Intermediate III-N1 from 3-chloro-5-oxo-6-phenyl-5, 6-dihydroimidazo [1, 2-c] pyrimidine-7-carbaldehyde using the corresponding reagents under appropriate conditions. MS (ESI) : mass calcd. for C₁₈H₂₁ClN₄O₂S is 392.90.

Intermediate III-P1: 2-methyl-N- (1- (1-oxo-2-phenyl-1, 2-dihydropyrrolo [1, 2-c] pyrimidin-3-yl) ethyl) propane-2-sulfinamide

Step 1： 2- (benzyloxycarbonylamino) -2-hydroxyacetic acid

To a mixture of benzyl carbamate (7.55 g, 50 mmol) in Et₂O (80 mL) was added 2-oxoacetic acid·1H₂O (5.05 g, 55 mmol) , the reaction was stirred at r.t. overnight. The mixture was concentrated in vacuo to afford the title compound as a white solid which was used in the next step without further purification.

Step 2: Methyl 2- (benzyloxycarbonylamino) -2-methoxyacetate

To a solution of 2- ( ( (benzyloxy) carbonyl) amino) -2-hydroxyacetic acid (about 11.25 g, 50 mmol) in MeOH (150 mL) was added concentrated sulfuric acid (2 mL) dropwise at 0 ℃. After the addition, the reaction mixture was stirred at r.t. for 90 h, then poured into the iced sat. NaHCO₃ aq. (300 mL) , the resulting mixture was extracted with EtOAc, the organic layers were dried over anhydrous Na₂SO₄, concentrated and purified by column chromatography to afford the title compound as a white solid (12 g) . Yeld: 95％. MS (ESI) : calcd. value for C₁₂H₁₅NO₅ is 253.25, m/z measured value is 275.7 (M+23) ⁺.

Step 3: Methyl 2- (benzyloxycarbonylamino) -2- (diethoxyphosphoryl) acetate

To a solution of methyl 2- ( ( (benzyloxy) carbonyl) amino) -2-methoxyacetate (12 g, 47.4 mmol) in toluene (60 mL) was added PBr₃ (12.8 g, 47.4 mmol) at 70 ℃, the reaction was stirred at 70 ℃ for 20 h, then triethyl phosphate (7.87 g, 47.4 mmol) was added dropwise and stirred at 70 ℃ for another 2 h. The mixture was concentrated, diluted with EtOAc, and washed with sat. NaHCO₃ aq.. The organic layers were dried over anhydrous Na₂SO₄, filtered and concentrated. The resulting residue was dissolved in EtOAc, petroleum ether was added with vigorous stirring, then filtrated to afford the title compound as a white solid (8 g) . Yield: 47％.

Step 4: Methyl 1-oxo-1, 2-dihydropyrrolo [1, 2-c] pyrimidine-3-carboxylate

To a solution of methyl 2- ( ( (benzyloxy) carbonyl) amino) -2- (diethoxyphosphoryl) acetate (8 g, 22.3 mmol) in DCM (80 mL) was added 1, 1, 3, 3-tetramethylguanidine (2.44 g, 21.2 mmol) at r.t., the reaction was stirred at r.t for 15min, then a solution of 1H-pyrrole-2-carbaldehyde (1.92 g, 20.2 mmol) in DCM (5 mL) was added dropwise at -30 ℃, the reaction mixture was stirred at -30 ℃ for 45 min, then warmed to r.t. and stirred for 48 h. The mixture was concentrated and purified by column chromatography to afford the title compound as a white solid (2 g) . Yield: 51％. MS (ESI) : calcd. value for C₉H₈N₂O₃ is 192.17, m/z measured value is 192.9 (M+1) ⁺.

Step 5: Methyl 1-oxo-2-phenyl-1, 2-dihydropyrrolo [1, 2-c] pyrimidine-3-carboxylate

To a solution of methyl 1-oxo-1, 2-dihydropyrrolo [1, 2-c] pyrimidine-3-carboxylate (576 mg, 3 mmol) in DCM (20 mL) was added phenylboronic acid (732 mg, 6 mmol) , copper (II) acetate (1.08 g, 6 mmol) , pyridine (1.18 g, 15 mmol) and

molecular sieve at r.t., the reaction was stirred at r.t. for 20h. The mixture was filtered, concentrated and purified by column chromatography to afford the title compound as a white solid (650 mg) . Yield: 81％. MS (ESI) : calcd. value for C₁₅H₁₂N₂O₃ is 268.27, m/z measured value is 268.8 (M+1) ⁺.

Step 6: 1-oxo-2-phenyl-1, 2-dihydropyrrolo [1, 2-c] pyrimidine-3-carboxylic acid

To a solution of methyl 1-oxo-2-phenyl-1, 2-dihydropyrrolo [1, 2-c] pyrimidine-3-carboxylate (1 g, 3.73 mmol) in EtOH (30 mL) and THF (30 mL) was added NaOH aq. (11.19 mL, 1N) at 0 ℃, the reaction was stirred at 0 ℃ for 30min. The mixture was concentrated, diluted with H₂O (10 mL) , adjusted to pH＝6 with HCl aq. (1N) and concentrated in vacuo to afford the title compound as a brown solid which was used in the next step without further purification. MS (ESI) : calcd. value for C₁₄H₁₀N₂O₃ is 254.25, m/z measured value is 254.7 (M+1) ⁺.

Step 7: N-methoxy-N-methyl-1-oxo-2-phenyl-1, 2-dihydropyrrolo [1, 2-c] pyrimidine -3-carboxamide

To a solution of 1-oxo-2-phenyl-1, 2-dihydropyrrolo [1, 2-c] pyrimidine-3-carboxylic acid (about 950 mg, 3.73 mmol) in DMF (10 mL) were added DIEA (1.44 g, 11.19 mmol) and HBTU (1.70 g, 4.48 mmol) , the mixture was stirred at r.t for 5 min, then N, O-dimethylhydroxylamine hydrochloride (438 mg, 4.48 mmol) was added, the reaction was stirred at r.t overnight. The mixture was concentrated and purified by column chromatography to afford the title compound as a white solid (550 mg) . Yield: 50％. MS (ESI) : calcd. value for C₁₆H₁₅N₃O₃ is 297.31, m/z measured value is 297.7 (M+1) ⁺.

Step 8: 3-acetyl-2-phenylpyrrolo [1, 2-c] pyrimidin-1 (2H) -one

To a solution of N-methoxy-N-methyl-1-oxo-2-phenyl-1, 2-dihydropyrrolo [1, 2-c] pyrimidine-3-carboxamide (550 mg, 1.85 mmol) in THF (5 mL) was added a solution of Methylmagnesium bromide in Et₂O (1.23 mL, 3N) at 0 ℃ under N₂, the reaction was stirred at 0 ℃ for 1h. The mixture was quenched with sat. NH₄Cl aq., concentrated and purified by column chromatography to afford the title compound as a yellow solid (220 mg) . Yield: 47％. MS (ESI) : calcd. value for C₁₅H₁₂N₂O₂ is 252.27, m/z measured value is 252.7 (M+1) ⁺.

Intermediate III-P1 was prepared according to the procedure of Intermediate III-N1 from 3-acetyl-2-phenylpyrrolo [1, 2-c] pyrimidin-1 (2H) -one using the corresponding reagents under appropriate conditions. MS (ESI) : calcd. value for C₁₉H₂₃N₃O₂S is 357.15, m/z measured value is 358.1 (M+1) ⁺.

Intermediate III-Q1: (S) -3- (1-aminoethyl) -8-chloro-2-phenylpyrrolo [1, 2-a] pyrazin-1 (2H) -one

Step 1: methyl 3-chloro-1- (2-oxopropyl) -1H-pyrrole-2-carboxylate

To a solution of methyl 3-chloro-1H-pyrrole-2-carboxylate (5.85 g, 36.7 mmol) in DMF (70 mL) was added 60％NaH (1.61 g, 40.3 mmol) at 0-5 ℃ and stirred at 0-5 ℃ for 30 minutes. Then a solution of 1-bromopropan-2-one (7.54 g, 55 mmol) in DMF (10 mL) was added dropwise at 0-5 ℃, and the reaction was stirred at room temperature for 30 minutes. After concentration in vacuo, the residue was used in the next step without further purification.

Step 2: 8-chloro-3-methyl-2-phenylpyrrolo [1, 2-a] pyrazin-1 (2H) -one

To a solution of aniline (53.9 mL, 591 mmol) in dichloromethane (500 mL) at 0-10 ℃was added dropwise trimethylaluminum (53.4 mL, 557 mmol) . The mixture was stirred for 1 h at room temperature and then methyl 3-chloro-1- (2-oxopropyl) -1H-pyrrole-2-carboxylate (50 g, 232 mmol) in dichloromethane was added dropwise. The reaction mixture was then stirred at reflux for 16 h and quenched slowly with water and 1 N HCl at 0-10 ℃. The layers were partitioned and the aqueous layer was extracted with dichloromethane. The combined organic layers were dried and concentrated to a residue. This residue was taken up in toluene (1 L) and p-TsOH was added. The mixture was stirred at reflux with employment of a Dean-Starks trap to remove water. After 20 h the reaction mixture was concentrated and purified by flash column chromatography (20％methanol + 20％ethyl acetate in petroleum ether) to provide the desired product in 78％yield.

Step 3: 8-chloro-1-oxo-2-phenyl-1, 2-dihydropyrrolo [1, 2-a] pyrazine-3-carbaldehyde

A solution of 8-chloro-3-methyl-2-phenylpyrrolo [1, 2-a] pyrazin-1 (2H) -one (10 g, 38.7 mmol) in dioxane (250 mL) was treated with selenium dioxide (10.94 g, 98.6 mmol) at room temperature. The mixture was stirred for 1 h at reflux and then filtered, concentrated, and purified by flash column chromatography (100％dichloromethane ramping to 95％dichloromethane/5％methanol) to provide the desired product in 57％yield.

Step 4: (R) -N- ( (8-chloro-1-oxo-2-phenyl-1, 2-dihydropyrrolo [1, 2-a] pyrazin-3-yl) methylene) -2-methylpropane-2-sulfinamide

A solution of 8-chloro-1-oxo-2-phenyl-1, 2-dihydropyrrolo [1, 2-a] pyrazine-3-carbaldehyde (800 mg, 2.93 mmol) in dioxane (8 mL) was treated with Ti (OEt) ₄ (5.87 mmol) and (R) -2-methylpropane-2-sulfinamide (2.93 mmol) at room temperature. The mixture was warmed to reflux for 2 h and then cooled to room temperature. Water (500 mL) and ethyl acetate (500 mL) were added. The organic layer was filtered and concentrated to provide the desired product in 91％yield.

Step 5: (R) -N- ( (S) -1- (8-chloro-1-oxo-2-phenyl-1, 2-dihydropyrrolo [1, 2-a] pyrazin-3-yl) ethyl) -2-methylpropane-2-sulfinamide

A solution of (R, E) -N- ( (8-chloro-1-oxo-2-phenyl-1, 2-dihydropyrrolo [1, 2-a] pyrazin-3-yl) methylene) -2-methylpropane-2-sulfinamide (38 g, 101 mmol) in dichloromethane (380 mL) was cooled to –78 ℃ and treated with MeMgCl (2.4 equiv) . The mixture was stirred for 2 h at room temperature, re-cooled to –78 ℃ and treated with additional MeMgCl (2.9 equiv) . The mixture was stirred for 1 h at room temperature and quenched with aqueous ammonium chloride. The layers were separated and the aqueous was extracted with dichloromethane three times. The combined organic layers were concentrated and purified through silica gel chromatography (5％methanol in dichloromethane) to provide a mixture of diastereomers (24 g, 3: 1) . The desired diastereomer (9.5 g) was isolated through SFC (OJ column, 70: 30 CO₂: EtOH with 0.1％ammonium hydroxide) .

Step 6: (S) -3- (1-aminoethyl) -8-chloro-2-phenylpyrrolo [1, 2-a] pyrazin-1 (2H) -one

To a solution of (R) -N- ( (S) -1- (8-chloro-1-oxo-2-phenyl-1, 2-dihydropyrrolo [1, 2-a] pyrazin-3-yl) ethyl) -2-methylpropane-2-sulfinamide (9.5 g, 33 mmol) in dichloromethane (95 mL) was added HCl in dioxane and methanol. The solution was stirred for 2 h at room temperature and then the pH adjusted to 7-8 with aqueous sodium bicarbonate. The layers were separated and the aqueous was washed with dichloromethane three times. The combined organics were concentrated under reduced pressure to provide the desired product (5.5 g, 77％) .

Intermediate III-A10: tert-butyl (S) -2- (5-chloro-3- (3-cyanophenyl) -4-oxo-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidine-1-carboxylate

A solution of tert-butyl (S) -2- (5-chloro-3- (3-iodophenyl) -4-oxo-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidine-1-carboxylate (Intermediate III-A7) (1.0 g, 1.9 mmol) and zinc cyanide (229 mg, 1.91 mmol) in DMF (9.5 mL) was degassed with nitrogen. Next, tetrakis (triphenylphosphine) palladium (0) (427 mg, 0.369 mmol) was added and the reaction heated to 80 ℃ for 18 hours. The reaction was partitioned between EtOAc and H₂O and the organic layer was washed with water and concentrated to dryness. The residue was purified by fast column chromatography to give the title compound (552 mg) . Yield: 68.3％. MS (ESI) : mass calcd. for C₂₁H₂₀ClN₅O₃, 425.87； m/z found, 326.0 [M-99] ⁺. ¹H NMR (400 MHz, CDCl₃) : δ 7.87 -7.78 (m, 1H) , 7.77 -7.58 (m, 2H) , 7.51 -7.33 (m, 2H) , 6.54 (d, J ＝ 2.8 Hz, 1H) , 4.69 -4.56 (m, 1H) , 3.94 (d, J ＝ 8.6 Hz, 1H) , 3.88 -3.75 (m, 1H) , 2.29 -2.11 (m, 2H) , 1.42 (s, 9H) .

Intermediate III-B5: tert-butyl (2S, 3S) -2- (5-chloro-3- (3- (methylsulfonyl) phenyl) -4-oxo-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidine-1-carboxylate

A solution of tert-butyl (2S, 3S) -2- (5-chloro-3- (3- (methylthio) phenyl) -4-oxo-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methyl-azetidine-1-carboxylate (Intermediate III-B4) (400 mg, 0.87 mmol) in DCM (4 mL) was treated with mCPBA (2.2 eq., 428 mg, 1.91 mmol) and was stirred for 19 h. The reaction was partitioned between DCM and saturated NaHCO₃. The DCM layer was collected, dried over anhydrous Na₂SO₄, and concentrated to dryness. The residue was purified by fast column chromatography to give the title compound (396 mg) . Yield: 92.6％. MS (ESI) : mass calcd. for C₂₂H₂₅ClN₄O₅S, 492.98； m/z found, 392.9 [M-99] ⁺. ¹H NMR (500 MHz, CDCl₃) : δ 8.11 (d, J ＝ 6.8 Hz, 1H) , 7.85 -7.44 (m, 3H) , 7.36 (d, J ＝ 2.9 Hz, 1H) , 6.54 (s, 1H) , 4.34 -3.99 (m, 2H) , 3.34 (dd, J ＝ 7.9, 4.5 Hz, 1H) , 3.11 (d, J ＝ 8.0 Hz, 3H) , 2.60 -2.46 (m, 1H) , 1.53-1.30 (m, 9H) , 0.91 (d, J ＝ 6.9 Hz, 3H) .

Example 1

Compound 1: 2- ( (2S, 3S) -1- (2-Amino-5, 5-dioxido-6, 7-dihydrothieno [3, 2-d] pyrimidin-4-yl) -3-methylazetidin-2-yl) -5-chloro-3-phenylpyrrolo [2, 1-f] [1, 2, 4] triazin-4 (3H) -one

Step 1.1: 5-Chloro-2- ( (2S, 3S) -3-methyl-1- (2- (methylthio) -6, 7-dihydrothieno [3, 2-d] pyrimidin-4-yl) azetidin-2-yl) -3-phenylpyrrolo [2, 1-f] [1, 2, 4] triazin-4 (3H) -one

To a solution of (2S, 3S) -tert-butyl 2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidine-1-carboxylate (Intermediate III-B1) (1.6 g, 3.86 mmol) in MeOH (5 mL) was added conc. HCl (5 mL) , the resulting mixture was stirred at rt. for 5 min, and then concentrated to dryness. The residue was dissolved in ethanol (20 mL) , then DIEA (1.9 mL, 11.58 mmol) and Intermediate I-1 (0.844 g, 3.86 mmol) were added. The resulting mixture was stirred at 90℃ for 24 h, and then concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (1.9 g) . Yield: 99％. MS (ESI) : calcd. value for C₂₃H₂₁ClN₆OS₂ is 496.1, m/z measured value is 497.1 (M+1) ⁺.

Step 1.2: 2- ( (2S, 3S) -1- (2-Amino-5, 5-dioxido-6, 7-dihydrothieno [3, 2-d] pyrimidin-4-yl) -3-methylazetidin-2-yl) -5-chloro-3-phenylpyrrolo [2, 1-f] [1, 2, 4] triazin-4 (3H) -one

To a solution of 5-chloro-2- ( (2S, 3S) -3-methyl-1- (2- (methylthio) -6, 7-dihydrothieno [3, 2-d] pyrimidin-4-yl) azetidin-2-yl) -3-phenylpyrrolo [2, 1-f] [1, 2, 4] triazin-4 (3H) -one (1.9 g, 3.82 mmol) in DCM (30 mL) was added m-CPBA (77％, 3.4g, 15.29 mmol) . The resulting mixture was stirred at rt. for 1h, and then purged with NH₃ for 7 min. The mixture was stirred at rt. overnight, and then poured into water, and extracted with DCM. The organic layers were washed sequentially with saturated NaHCO₃ solution, Na₂SO₃ solution and water, and then concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (1.2 g) . Yield: 65％. ¹H NMR (400 MHz, CD₃OD) δ 7.73-7.65 (m, 1H) , 7.62-7.53 (m, 3H) , 7.35 (d, J ＝ 6.0 Hz, 2H) , 6.50 (d, J ＝2.8 Hz, 1H) , 4.77 (brs, 1H) , 4.46 (brs, 1H) , 3.59 (brs, 1H) , 3.43 –3.39 (m, 2H) , 3.17-3.07 (m, 2H) , 2.66 (brs, 1H) , 0.75 (d, J ＝ 6.8 Hz, 3H) . MS (ESI) : calcd. value for C₂₂H₂₀ClN₇O₃S is 497.1, m/z measured value is 498.3 (M+1) ⁺.

The following compounds were prepared according to the procedure of Compound 1 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:

Example 2

Compound 16: (S) -2-Amino-4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [1, 2-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6, 7-dihydro-5H-pyrrolo [3, 4-d] pyrimidin-5-one

Step 2.1: (S) -tert-Butyl 2-amino-4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [1, 2-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) -carboxylate

To a solution of Intermediate I-2 (240 mg, 0.81 mmol) in DCM (10 mL) was added m-CPBA (77％, 237 mg, 1 mmol) , the resulting mixture was stirred at rt. for 1h, and used in next step directly.

To a solution of Intermediate III-A1 (357 mg, 0.89 mmol) in MeOH (4 mL) was added conc. HCl (2 mL) . The resulting mixture was stirred at rt. for 0.5h, and then concentrated to dryness. The residue was dissolved in EtOH (5 mL) , and added DIEA (313 mg, 2.43 mmol) , followed by the above reaction mixture of Intermediate I-2. The reaction mixture was stirred at rt. for 1h, then added aq. Na₂S₂O₃. The mixture was concentrated to dryness and purified by flash column chromatography to afford the title compound as a solid (330 mg) . Yield: 74％. MS (ESI) : calcd. value for C₂₆H₂₅ClN₈O₄ is 548.17, m/z measured value is 549.2 (M+1) ⁺.

Step 2.2: (S) -2-Amino-4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [1, 2-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6, 7-dihydro-5H-pyrrolo [3, 4-d] pyrimidin-5-one

The mixture of (S) -tert-butyl 2-amino-4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [1, 2-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) -carboxylate (330 mg, 0.6 mmol) in TFA (4 mL) was stirred at rt. for 0.5h. The mixture was concentrated to dryness, the resultant residue was dissolved in EtOAc, neutralized with aq. NaHCO₃ to pH＝7, and then concentrated to dryness. and purified by flash column chromatography to afford the title compound as a solid (230 mg) . Yield: 85％. ¹H NMR (400 MHz, CD₃OD) δ 7.71 (d, J ＝ 7.2 Hz, 1H) , 7.60 –7.50 (m, 3H) , 7.35 –7.24 (m, 2H) , 6.47 (brs, 1H) , 5.43 (brs, 1H) , 4.24 –4.12 (m, 1H) , 4.07 (s, 2H) , 3.97 (brs, 1H) , 2.45 –2.31 (m, 1H) , 2.30 –2.15 (m, 1H) . MS (ESI) : calcd. value for C₂₁H₁₇ClN₈O₂ is 448.87, m/z measured value is 449.0 (M+1) ⁺.

The following compounds were prepared according to the procedure of Compound 16 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:

Example 3

Compound 7: (S) -2-Amino-4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [1, 2-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6-methylpyrido [4, 3-d] pyrimidin-5 (6H) -one

Step 3.1: (S) -Ethyl 4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [1, 2-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6-methyl-2- (methylthio) pyrimidine-5-carboxylate

The mixture of intermediate III-A1 (1 g, 2.5 mmol) in conc. HCl (2 mL) and MeOH (4 mL) was stirred at rt. for 30 min, and then concentrated. The resulting residue was dissolved in EtOH (20 mL) and DIEA (0.97 g, 7.5 mmol) , and then ethyl 4-chloro-6-methyl-2- (methylthio) pyrimidine-5-carboxylate (1 g, 2.5 mmol) was added. The mixture was stirred at rt. for 1 h, and then concentrated to dryness and purified by flash column chromatography to afford the title compound as a solid (1.23 g) . Yield: 96％. MS (ESI) : calcd. value for C₂₄H₂₃ClN₆O₃S is 510.12, m/z measured value is 511.1 (M+1) ⁺.

Step 3.2: (S) -Ethyl 4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [1, 2-f] [1, 2, 4] triazin -2-yl) azetidin-1-yl) -6- (2- (dimethylamino) vinyl) -2- (methylthio) pyrimidine-5-carboxylate

To a solution of (S) -ethyl 4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [1, 2-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6-methyl-2- (methylthio) pyrimidine-5-carboxylate (1.23 g, 2.41 mmol) in DMF (10 mL) was added 1-tert-butoxy-N, N, N' , N' -tetramethylmethanediamine (838 mg, 4.82 mmol) . The resulting mixture was stirred at 100℃ overnight, and then concentrated to dryness to afford the title compound as a crude product (1.36 g) , which was used for next step directly.

Step 3.3: (S) -4- (2- (5-Chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [1, 2-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6-methyl-2- (methylthio) pyrido [4, 3-d] pyrimidin-5 (6H) -one

To a suspension of the above crude product (1.36 g, about 2.41 mmol) in EtOH (15 mL) was added methanamine/EtOH solution (28％, 10 mL) . The mixture was stirred at reflux overnight, and then concentrated to remove some solvent. The precipitate was filtered to afford the title compound as a solid (930 mg) . Yield for two steps: 76％. MS (ESI) : calcd. value for C₂₄H₂₀ClN₇O₂S is 505.11, m/z measured value is 506.0 (M+1) ⁺.

Step 3.4: (S) -2-Amino-4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [1, 2-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6-methylpyrido [4, 3-d] pyrimidin-5 (6H) -one

To a solution of (S) -4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [1, 2-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6-methyl-2- (methylthio) pyrido [4, 3-d] pyrimidin-5 (6H) -one (870 mg, 1.72 mmol) in DCM (30 mL) was added m-CPBA (77％, 811 mg, 3.61 mmol) . The resulting mixture was stirred at rt. for 3 h, and then bubbled with NH₃ for 5 min, and then concentrated to dryness. The residue was dissolved in dioxane (20 mL) and NH₃. H₂O (7 mL) . The mixture was stirred at 100℃ for 4 h, and then concentrated to dryness and purified by flash column chromatography to afford the title compound as a solid (550 mg) . Yield: 67％. ¹H NMR (400 MHz, DMSO-d₆) δ 7.78 –7.34 (m, 7H) , 6.63 -6.50 (m, 3H) , 5.95-5.92 (m, 1H) , 5.52 (brs, 0.3H) , 4.75 (brs, 0.7H) , 4.61 (brs, 0.7H) , 4.12 (brs, 0.3H) , 3.81 (brs, 0.3H) , 3.59 (brs, 0.7H) , 3.19 (s, 3H) , 2.47 -2.30 (m, 1H) , 2.09 (brs, 0.3H) , 1.75 (brs, 0.7H) . MS (ESI) : calcd. value for C₂₃H₁₉ClN₈O₂ is 474.13, m/z measured value is 475.1 (M+1) ⁺.

Example 4

Compound 131: (S) -4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -2- (methylamino) -6, 7-dihydro-5H-pyrrolo [3, 4-d] pyrimidin-5-one

Step 4.1: tert-butyl 4- (ethylsulfinyl) -2- (methylsulfinyl) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) -carboxylate

To a solution of intermediate I-8 (68 mg, 0.2 mmol) in DCM (6 mL) was added m-CPBA (98 mg, 0.44 mmol) . The mixture was stirred at rt. for 1 h, and then used for the next step directly. MS (ESI) : calcd. value for C₁₄H₁₉N₃O₅S₂ is 373.08, m/z measured value is 746.7 (2M+1) ⁺.

Step 4.2: (S) -tert-butyl 4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -2- (methylamino) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) -carboxylate

To a solution of intermediate II-1 (80 mg, 0.2 mmol) in MeOH (2 mL) was added conc. HCl (2 mL) . The mixture was concentrated to dryness. The residue was dissolved in dioxane (4 mL) , and then DIEA (78 mg, 0.6 mmol) was added, followed by the above reaction mixture of step 4.1. The resulting mixture was stirred at rt. for 3 h, and then methanamine /EtOH solution (1 mL, 27％-32％) was added. The mixture was stirred at rt. overnight. After the reaction was completed, the mixture was diluted with EtOAc and saturated NaHCO₃ solution. The organic layer was dried over Na₂SO₄, filtered, and concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (45 mg) . Yield: 40％. MS (ESI) : calcd. value for C₂₇H₂₇ClN₈O₄ is 562.18, m/z measured value is 563.3 (M+1) ⁺.

Step4.3: (S) -4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -2- (methylamino) -6, 7-dihydro-5H-pyrrolo [3, 4-d] pyrimidin-5-one

The mixture of (S) -tert-butyl 4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -2- (methylamino) -5-oxo-5H-pyrrolo [3, 4-d] pyrimidine-6 (7H) - carboxylate (45 mg, 0.08 mmol) in TFA (1 mL) was evaporated to dryness. The residue was dissolved in MeOH (2 mL) and neutralized to pH＝7-8 with NaHCO₃. The mixture was filtered, the filtrate was concentrated to dryness and purified by flash column chromatography to afford the title compound as a solid (25 mg) . Yield: 68％. ¹H NMR (400 MHz, CD₃OD) δ 7.71 (brs, 1H) , 7.64 –7.49 (m, 3H) , 7.36 –7.25 (m, 2H) , 6.51 (brs, 1H) , 5.41 (brs, 1H) , 4.19 (brs, 1H) , 4.08 (s, 2H) , 4.04 –3.94 (m, 1H) , 2.90 (s, 3H) , 2.38 (brs, 1H) , 2.24 (brs, 1H) . MS (ESI) : calcd. value for C₂₂H₁₉ClN₈O₂ is 462.13, m/z measured value is 463.2 (M+1) ⁺.

The following compounds were prepared according to the procedure of Compound 131 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:

Example 5

Compound 123: 6-Amino-4- ( (2S, 3S) -2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidin-1-yl) -1H-pyrazolo [3, 4-d] pyrimidin-3 (2H) -one

Step 5.1: Ethyl 4-chloro-6- ( (2S, 3S) -2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2， 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidin-1-yl) -2- (methylthio) pyrimidine-5-carboxylate.

To a solution of intermediate III-B1 (124 mg, 0.3 mmol) in MeOH (2 mL) was added 1 mL of conc. HCl dropwise. The mixture was concentrated to dryness, the residue was dissolved in EtOH (5 mL) , and then DIEA (116 mg, 0.9 mmol) and ethyl 4, 6-dichloro-2- (methylthio) pyrimidine-5-carboxylate (80 mg, 0.3 mmol) were added, the resulting mixture was stirred at rt. for 3h. After the reaction was completed, the mixture was concentrated to dryness and purified by flash column chromatography to afford the title compound as a solid (163 mg) . MS (ESI) : calcd. value for C₂₄H₂₂Cl₂N₆O₃S is 544.1, m/z measured value is 545.3 (M+1) ⁺.

Step 5.2: 4- ( (2S, 3S) -2- (5-Chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidin-1-yl) -6- (methylthio) -1H-pyrazolo [3, 4-d] pyrimidin-3 (2H) -one

To a solution of ethyl 4-chloro-6- ( (2S, 3S) -2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidin-1-yl) -2- (methylthio) pyrimidine-5-carboxylate (163 mg, 0.3 mmol) in EtOH (10 mL) were added NH₂NH₂. H₂SO₄ (40 mg, 0.3 mmol) and DIEA (3 mL) , the resulting mixture was stirred at 100℃ overnight, and then concentrated to dryness. The residue was purified by flash column chromatography to afford the title compound as a solid (148 mg) . MS (ESI) : calcd. value for C₂₂H₁₉ClN₈O₂S is 494.1, m/z measured value is 495.2 (M+1) ⁺.

Step 5.3: 6-Amino-4- ( (2S, 3S) -2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidin-1-yl) -1H-pyrazolo [3, 4-d] pyrimidin-3 (2H) -one

To a solution of 4- ( (2S, 3S) -2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidin-1-yl) -6- (methylthio) -1H-pyrazolo [3, 4-d] pyrimidin-3 (2H) -one (148 mg, 0.3 mmol) in dry DCM (5 mL) was added m-CPBA (87 mg, 0.39 mmol) , the resulting mixture was stirred at rt. for 0.5 h, and then concentrated to dryness. The residue was dissolved in dioxane (10 mL) , and bubbled with NH₃ for 10 min, then sealed and heated at 100℃ overnight. After reaction was completed, the mixture was concentrated to dryness and purified by flash column chromatography to afford the title compound as a solid (15 mg) . Yield: 10.8％. ¹H NMR (400 MHz, DMSO-d₆) δ 7.90 (d, J ＝ 8.0 Hz, 1H) , 7.78-7.73 (m, 1H) , 7.56-7.50 (m, 4H) , 7.48-7.45 (m, 1H) , 7.31-7.24 (m, 1H) , 6.64-6.44 (m, 3H) , 5.04 (d, J ＝ 2.8 Hz, 1H) , 4.12-4.08 (m, 1H) , 2.65-2.52 (m, 1H) , 2.00-1.93 (m, 1H) , 0.49 (d, J ＝ 6.4 Hz, 3H) . MS (ESI) : calcd. value for C₂₁H₁₈ClN₉O₂ is 463.1, m/z measured value is 464.2 (M+1) ⁺.

The following compound was prepared according to the procedure of Compound 123 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:

Example 6

Compound 122: 4- ( (2S, 3S) -2- (5-Chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidin-1-yl) -1H-pyrazolo [3, 4-d] pyrimidin-3 (2H) -one

Step 6.1: Ethyl 4-chloro-6- ( (2S, 3S) -2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidin-1-yl) pyrimidine-5-carboxylate

To a solution of Intermediate III-B1 (82 mg, 0.2 mmol) in MeOH (2 mL) was added 1 mL of conc. HCl, the mixture was concentrated. The residue was dissolved in EtOH (5 mL) , and then added DIEA (78 mg, 0.6 mmol) and ethyl 4, 6-dichloropyrimidine-5-carboxylate (44 mg, 0.2 mmol) , the resulting mixture was stirred at rt. for 2 h. After the reaction was completed, the mixture was concentrated and purified by flash column chromatography to afford the title compound as a solid (99 mg) . MS (ESI) : calcd. value for C₂₃H₂₀Cl₂N₆O₃ is 498.1, m/z measured value is 499.3 (M+1) ⁺.

Step 6.2: 4- ( (2S, 3S) -2- (5-Chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidin-1-yl) -1H-pyrazolo [3, 4-d] pyrimidin-3 (2H) -one

To a solution of ethyl 4-chloro-6- ( (2S, 3S) -2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidin-1-yl) pyrimidine-5-carboxylate (99 mg， 0.2 mmol) in EtOH (10 mL) were added NH₂NH₂. H₂SO₄ (52 mg, 0.4 mmol) and DIEA (52 mg, 0.4 mmol) , the resulting mixture was stirred at 100℃ overnight. After the reaction was completed, the mixture was concentrated to dryness and purified by flash column chromatography to afford the title compound as a solid (80 mg) . Yield: 98％. ¹H NMR (400 MHz, CD₃OD) δ 7.99 (s, 1H) , 7.82 –7.75 (m, 1H) , 7.64-7.53 (m, 3H) , 7.41-7.38 (m, 1H) , 7.21 (brs, 1H) , 6.44 (s, 1H) , 5.33 (brs, 1H) , 4.40-4.35 (m, 1H) , 3.63-3.55 (m, 1H) , 2.59 (brs, 1H) , 0.63 (d, J ＝ 6.8 Hz, 3H) . MS (ESI) : calcd. value for C₂₁H₁₇ClN₈O₂ is 448.1, m/z measured value is 449.2 (M+1) ⁺.

The following compound was prepared according to the procedure of Compound 122 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:

Example 7

Compound 2: (S) -2-amino-4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6-methyl-6, 7-dihydro-5H-pyrrolo [3, 4-d] pyrimidin-5-one

Step 7.1: (S) -ethyl 4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6-methyl-2- (methylthio) pyrimidine-5-carboxylate

To a solution of Intermediate III-A1 (400 mg, 1 mmol) dissolved in MeOH (4 mL) was added conc. HCl (4 mL) . The resulting mixture was concentrated at 50℃ to dryness. The residue was dissolved in n-BuOH (20 mL) , and then added DIEA (1 mL) and ethyl 4-chloro-6-methyl-2- (methylthio) pyrimidine-5-carboxylate (246 mg, 1 mmol) . The mixture was stirred at rt. for 2h, and then concentrated to remove the solvent. The residue was purified by flash column chromatography to afford the title compound as a solid (500 mg) . Yield: 97.8％. MS (ESI) : calcd. value for C₂₄H₂₃ClN₆O₃S is 510.12, m/z measured value is 511.1 (M+1) ⁺.

Step 7.2: (S) -4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6-methyl-2- (methylthio) -6, 7-dihydro-5H-pyrrolo [3, 4-d] pyrimidin-5-one

To a solution of (S) -ethyl4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6-methyl-2- (methylthio) pyrimidine-5-carboxylate (102 mg, 0.2 mmol) dissolved in dioxane (20 mL) , was added SeO₂ (110 mg, 1 mmol) . The mixture was stirred at reflux overnight, and then filtered. The filtrate was concentrated to dryness. The resulting residue was dissolved in DCM (1 mL) and MeOH (1 mL) , methanamine (31 mg, 1 mmol) was added. The mixture was stirred at rt. for 30 min, NaBH₃CN (62mg, 1mmol) was added. The reaction mixture was stirred at rt. overnight, and then concentrated to dryness and purified by flash column chromatography to afford the title compound as a solid (80 mg) . Yield: 81.6％. MS (ESI) : calcd. value for C₂₃H₂₀ClN₇O₂S is 493.11, m/z measured value is 494.2 (M+1) ⁺.

Step 7.3: (S) -2-amino-4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6-methyl-6, 7-dihydro-5H-pyrrolo [3, 4-d] pyrimidin-5-one

To a solution of (S) -4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6-methyl-2- (methylthio) -6, 7-dihydro-5H-pyrrolo [3, 4-d] pyrimidin-5-one (80 mg, 0.16 mmol) dissolved in DCM (10 mL) , was added m-CPBA (70％, 200 mg, 0.8 mmol) . The resulting mixture was stirred at rt. for 30 min, and then bubbled with NH₃ for a while. The resulting precipitate was filtered, and the filtrated was concentrated to dryness. The residue was dissolved in NH₃ /dioxane (0.5 M, 20 mL) , the mixture was stirred at 130℃ in a sealed tube overnight. The mixture was concentrated to dryness and purified by flash column chromatography to afford the title compound as a solid (40mg) . Yield: 53％. ¹H NMR (400 MHz, CD₃OD) δ 7.78 (s, 1H) , 7.58 –7.50 (m, 3H) , 7.31 –7.28 (m, 2H) , 6.47 (s, 1H) , 5.44 (brs, 1H) , 4.15 –3.97 (m, 4H) , 2.92 (s, 3H) , 2.34 (brs, 1H) , 2.20 -2.15 (m, 1H) . MS (ESI) : calcd. value for C₂₂H₁₉ClN₈O₂ is 462.13, m/z measured value is 463.3 (M+1) ⁺.

Example 8

Compound 171: (S) -5-chloro-2- (1- (5-hydroxythieno [2, 3-d] pyrimidin-4-yl) azetidin-2-yl) -3-phenylpyrrolo [2, 1-f] [1, 2, 4] triazin-4 (3H) -one

Step 8.1: Ethyl 4-chloro-6- ( (2-methoxy-2-oxoethyl) thio) pyrimidine-5-carboxylate

To a solution of ethyl 4, 6-dichloropyrimidine-5-carboxylate (663 mg, 3 mmol) in CH₃CN (15 mL) was added methyl 2-mercaptoacetate (318 mg, 3 mmol) and K₂CO₃ (414 mg, 3 mmol) . The resulting mixture was stirred at r.t. for 2 h, the mixture was diluted with EA, the organic layer was washed with H₂O, brine and dried over Na₂SO₄. The mixture was concentrated to dryness to afford the title compound as oil (900mg) . Yield: 100％. MS (ESI) : calcd. value for C₁₀H₁₁ClN₂O₄S is 290.01, m/z measured value is 290.9 (M+1) ⁺.

Step 8.2: Methyl 4-chloro-5-oxo-5, 6-dihydrothieno [2, 3-d] pyrimidine-6-carboxylate

To a solution of Ethyl 4-chloro-6- ( (2-methoxy-2-oxoethyl) thio) pyrimidine-5-carboxylate (900 mg, 3 mmol) in MeOH (30 mL) was added NaOMe (405 mg, 7.5 mmol) . The resulting mixture was stirred at r.t. overnight, the precipitate was collected by filtration to afford crude title compound as a solid (700mg) . Yield: 95％. MS (ESI) : calcd. value for C₈H₅ClN₂O₃S is 244.65, m/z measured value is 244.9 (M+1) ⁺.

Step 8.3: Methyl 4- ( (S) -2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -5-oxo-5, 6-dihydrothieno [2, 3-d] pyrimidine-6-carboxylate

The mixture of Intermediate III-A1 (200 mg, 0.5 mmol) in con. HCl (2 mL) and MeOH (4 mL) was stirred at r.t. for 30min, the mixture was concentrated to dryness, the residue was dissolved in EtOH (10 mL) , DIEA (194 mg, 1.5 mmol) was added followed by methyl 4-chloro-5-oxo-5, 6-dihydrothieno [2, 3-d] pyrimidine-6-carboxylate (122 mg, 0.5 mmol) , the resulting mixture was stirred at r.t. overnight. The mixture was concentrated to dryness and the residue was purified by flash column chromatography to afford the title compound as a solid (230mg) . Yield: 90％. MS (ESI) : calcd. value for C₂₃H₁₇ClN₆O₄S is 508.07, m/z measured value is 509.1 (M+1) ⁺.

Step 8.4: (S) -5-chloro-2- (1- (5-hydroxythieno [2, 3-d] pyrimidin-4-yl) azetidin-2-yl) -3-phenyl pyrrolo [2, 1-f] [1, 2, 4] triazin-4 (3H) -one

To a solution of Methyl 4- ( (S) -2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -5-oxo-5, 6-dihydrothieno [2, 3-d] pyrimidine-6-carboxylate (100 mg, 0.2 mmol) in DMSO (3 mL) was added NaCl (200 mg) and H₂O (0.1 mL) . The resulting mixture was stirred at 150℃ for 2 h. The mixture was cooled to r.t. and diluted with EA, the organic layer was washed with H₂O and brine, dried over Na₂SO₄ and concentrated to dryness, the residue was purified by preparative-TLC to give the title compound as a solid (15 mg) . Yield: 17％. ¹H NMR (400 MHz, CD₃OD) δ 8.33 (s, 0.7H) , 8.28 (s, 0.3H) , 7.76 (d, J ＝ 8.0 Hz, 0.7H) , 7.64 –7.55 (m, 4H) , 7.41 (brs, 0.3H) , 7.35 –7.31 (m, 1.3H) , 7.28 (d, J ＝ 3.2 Hz, 0.7H) , 6.54 –6.52 (m, 0.3H) , 6.50 (d, J ＝ 3.2 Hz, 0.7H) , 5.67 –5.64 (m, 1H) , 4.33 –4.26 (m, 1H) , 4.12 –4.05 (m, 1H) , 2.47 –2.39 (m, 1H) , 2.35 –2.26 (m, 1H) . MS (ESI) : calcd. value for C₂₁H₁₅ClN₆O₂S is 450.07, m/z measured value is 451.3 (M+1) ⁺.

Example 9

Compound 172: (S) -2-amino-4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) thieno [2, 3-d] pyrimidin-5 (6H) -one

Step 9.1: (S) -ethyl 4-chloro-6- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -2- (methylthio) pyrimidine-5-carboxylate

Intermediate III-A1 (2 g, 5 mmol) was dissolved in con. HCl (4 mL) and MeOH (8 mL) , the mixture was stirred at r.t. for 30 min, concentrated to dryness, the residue was dissolved in EtOH (40 mL) , DIEA (1.9 g, 15 mmol) was added followed by ethyl 4, 6-dichloro-2- (methylthio) pyrimidine-5-carboxylate (1.3 g, 5 mmol) . The resulting mixture was stirred at r.t. for 2 h, the precipitate was collected by filtration to afford the title compound as a solid (2 g) . Yield: 75％. MS (ESI) : calcd. value for C₂₃H₂₀Cl₂N₆O₃S is 530.07, m/z measured value is 531.1 (M+1) ⁺.

Step 9.2: (S) -ethyl 2-amino-4-chloro-6- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) pyrimidine-5-carboxylate

To a solution of (S) -ethyl 4-chloro-6- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -2- (methylthio) pyrimidine-5-carboxylate (420 mg, 0.79 mmol) in DCM (40 mL) was added m-CPBA (77％) (355 mg, 1.58 mmol) . The resulting mixture was stirred at r.t. for 2 h, then bubbled with NH₃ at 0℃ and stired at 0℃for 30 min, the mixture was diluted with DCM, the organic layer was washed with aq.Na₂S₂O₃, aq. NaHCO₃ and brine, dried over Na₂SO₄. The organic layer was concentrated to dryness and purified by flash column chromatography to afford the title compound as a solid (260 mg) . Yield: 66％. MS (ESI) : calcd. value for C₂₂H₁₉Cl₂N₇O₃ is 499.09, m/z measured value is 500.4 (M+1) ⁺.

Step 9.3: (S) -ethyl 2-amino-4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6- ( (2-methoxy-2-oxoethyl) thio) pyrimidine-5-carboxylate

To a solution of (S) -ethyl 2-amino-4-chloro-6- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) pyrimidine-5-carboxylate (260 mg, 0.52 mmol) in CH₃CN (10 mL) was added methyl 2-mercaptoacetate (83 mg, 0.78 mmol) and K₂CO₃ (108 mg, 0.78 mmol) . The resulting mixture was stirred at 100℃ for 4 h. The mixture was diluted with EA, the organic layer was washed with H₂O, brine, dried over Na₂SO₄ and concentrated to dryness to afford the crude title compound as an oil (350 mg) . It was used in next step without purification. MS (ESI) : calcd. value for C₂₅H₂₄ClN₇O₅S is 569.12, m/z measured value is 570.2 (M+1) ⁺.

Step 9.4: methyl 2-amino-4- ( (S) -2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -5-oxo-5, 6-dihydrothieno [2, 3-d] pyrimidine-6-carboxylate

To a solution of (S) -ethyl 2-amino-4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6- ( (2-methoxy-2-oxoethyl) thio) pyrimidine-5-carboxylate (350 mg, 0.52 mmol) in MeOH (20 mL) was added NaOMe (140 mg, 2.6 mmol) . The resulting mixture was stirred at r.t. overnight. The mixture was poured into H₂O, extracted with EA, the organic layer was washed with H₂O, brine, dried over Na₂SO₄ and concentrated to dryness to give the crude title compound as a solid (300 mg) . It was used in next step without purification. MS (ESI) : calcd. value for C₂₃H₁₈ClN₇O₄S is 523.08, m/z measured value is 524.2 (M+1) ⁺.

Step 9.5: (S) -2-amino-4- (2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) thieno [2, 3-d] pyrimidin-5 (6H) -one

To a solution of methyl 2-amino-4- ( (S) -2- (5-chloro-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -5-oxo-5, 6-dihydrothieno [2, 3-d] pyrimidine-6-carboxylate (300 mg, 0.52 mmol) in DMSO (40 mL) was added NaCl (500 mg) and H₂O (0.12 mL) . The resulting mixture was stirred at 150℃ for 3 h. The mixture was poured into H₂O, extracted with EA, the organic layer was washed with H₂O, brine, dried over Na₂SO₄, concentrated to dryness and purified by flash column chromatography to afford the title compound as a solid (50 mg) . Yield: 21％. ¹H NMR (400 MHz, DMSO-d₆) δ 7.74 –7.68 (m, 1H) , 7.61 –7.50 (m, 4H) , 7.42 –7.38 (m, 1H) , 7.22 (s, 1.5H) , 7.11 (brs, 0.5H) , 6.64 (d, J ＝2.4 Hz, 0.4H) , 6.59 (d, J ＝ 2.8 Hz, 0.6H) , 5.38 –5.34 (m, 0.6H) , 4.77 –4.73 (m, 0.4H) , 4.51 –4.44 (m, 0.4H) , 4.06 –4.02 (m, 0.6H) , 3.93 –3.83 (m, 1H) , 3.73 –3.61 (m, 0.8H) , 3.58 (s, 1.2H) , 2.47 –2.39 (m, 1H) , 2.14 –2.08 (m, 0.5H) , 2.01 –1.88 (m, 0.5H) . MS (ESI) : calcd. value for C₂₁H₁₆ClN₇O₂S is 465.08, m/z measured value is 466.2 (M+1) ⁺.

The following compound was prepared according to the procedure of Compound 172 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:

Example 10

Compound 188: 2-amino-4- ( (2S, 3S) -2- (5- (hydroxymethyl) -4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidin-1-yl) -6, 7-dihydro-5H-pyrrolo [3, 4-d] pyrimidin-5-one

To a solution of tert-butyl 2-amino-4- ( (2S, 3S) -2- (5- (methoxymethyl) -4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidin-1-yl) -5-oxo-5, 7-dihydro-6H-pyrrolo [3, 4-d] pyrimidine-6-carboxylate (62 mg, 0.1 mmol) in DCM (1.5 mL) was added TFA (3.0 mL) at 0℃, and stirred at r.t. for 40 min, then concentrated at r.t.. The residue was dissolved in MeOH (3.0 mL) , and treated with 1N LiOH (3.0 mL) , then stirred at r.t. for another 1 hour. The mixture was adjusted to pH ＝ 6.0～7.0, then concentrated to dryness and purified by fast column chromatography to afford the title compound as a white solid (5 mg) . Yield: 11％. ¹H NMR (400 MHz, CD₃OD) δ 7.78 (d, J ＝ 6.8 Hz, 1H) , 7.68 –7.53 (m, 3H) , 7.36-7.33 (m, 1H) , 7.23 (brs, 1H) , 6.53 (brs, 1H) , 5.19 (brs, 1H) , 4.84 (s, 2H) , 4.32-4.28 (m, 1H) , 4.08 (s, 2H) , 3.52 (brs, 1H) , 2.53 (brs, 1H) , 0.64 (d, J ＝ 6.8 Hz, 3H) . MS (ESI) : calcd. value for C₂₃H₂₂N₈O₃ is 458.48, m/z measured value is 459.2 (M+1) ⁺.

Example 11

Compound 190: 2-amino-4- ( (2S, 3S) -3-methyl-2- (4-oxo-3, 5-diphenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) azetidin-1-yl) -6, 7-dihydro-5H-pyrrolo [3, 4-d] pyrimidin-5-one

2-amino-4- ( (2S, 3S) -2- (5-bromo-4-oxo-3-phenyl-3, 4-dihydropyrrolo [2, 1-f] [1, 2, 4] triazin-2-yl) -3-methylazetidin-1-yl) -6, 7-dihydro-5H-pyrrolo [3, 4-d] pyrimidin-5-one (45 mg, 0.09 mmol) , phenylboronic acid (22 mg, 0.18 mmol) , Na₂CO₃ (29 mg, 0.27 mmol) and Pd(dppf) Cl₂·CH₂Cl₂ (8 mg, 0.009 mmol) were stirred in a tube under N₂, then a mixture of dioxane (10 mL) and H₂O (1 mL) were added. The tube was bubbled with N₂ for 7 min and sealed. The mixture was irradiated in the microwave at 110 ℃ for 2 h. The mixture was then concentrated and purified by fast column chromatography to afford the title compound as a yellow solid (12 mg) . Yield: 26％. ¹H NMR (400 MHz, CD₃OD) δ 7.79 (d, J ＝ 7.6 Hz, 1H) , 7.67 (d, J ＝ 7.6 Hz, 2H) , 7.62 –7.51 (m, 3H) , 7.38 –7.26 (m, 4H) , 7.23-7.19 (m, 1H) , 6.66 (brs, 1H) , 5.21 (brs, 1H) , 4.35-4.31 (m, 1H) , 4.11 (s, 2H) , 3.54 (br, 1H) , 2.56 (br, 1H) , 0.65 (d, J ＝ 6.4 Hz, 3H) . MS (ESI) : calcd. value for C₂₈H₂₄N₈O₂ is 504.55, m/z measured value is 505.4 (M+1) ⁺.

The following compound was prepared according to the procedure of Compound 190 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:

Example 12

Kinase Inhibition assays of p110α/p85α, p110β/p85α, p110δ/p85α and p110γ

PI3K kinases including p110α/p85α and p110γ were purchased from Invitrogen, p110δ/p85α and p110β/p85α were from Millipore.

Primary screening data and IC₅₀ values were measured using Transcreener^TM KINASE Assay (Bellbrook, Catalog # 3003-10K) . The Assay can be carried out according to the procedures suggested by the manufacturer. It is a universal, homogenous, high throughput screening (HTS) technology using a far-red, competitive fluorescence polarization immunoassay based on the detection of ADP to monitor the activity of enzymes that catalyze group transfer reactions. Briefly, the Transcreener KINASE Assay is designed as a simple two-part, endpoint assay as follows:

1) Preparation of 25uL kinase reaction: the 25uL kinase reaction is performed by preparing a reaction mixture containing 10uL kinase buffer (50mM HEPES, 100mM NaCl, 1 mM EGTA, 0.03％CHAPS, 3mM MgCl₂, and freshly supplemented 1mM DTT) , and 10uL 30uM PIP2 and10uM ATP, 5uL test compound solution (the compound was dissolved in DMSO, the final concentrations of the compound in the reaction mixture are at 1 uM, 0.3uM, 0.1uM, 0.037uM, 0.012uM, 0.0041 uM, 0.0014uM and 0.0005uM, and final concentration of DMSO in the reaction mixture was 2％) or 5uL control (2％DMSO) . The reaction mixture was added into desired wells of a 96-well plate. The plate was sealed and incubated for 80min at room temperature.

2) Next, 25uL ADP detection mix was added into each well. The plate was sealed again and incubated for 60min at room temperature. Then fluorescence polarization was measured by Tecan Infinite F500 Reader.

Data was analyzed and IC₅₀ values were generated using the add-in software for Microsoft Excel, Xlfit^TM (version 5.3) .

Inhibition rates were calculated as follow: IH％＝ (ADP amount under 2％DMSO well-ADP amount under test compound well) /ADP amount under 2％DMSO well × 100％.

In vitro activity data:

The foregoing description of the specific embodiments will so fully reveal the general nature of the invention that others can, by applying knowledge within the skill of the art, readily modify and/or adapt for various applications such specific embodiments, without undue experimentation, without departing from the general concept of the present invention. Therefore, such adaptations and modifications are intended to be within the meaning and range of equivalents of the disclosed embodiments, based on the teaching and guidance presented herein. It is to be understood that the phraseology or terminology herein is for the purpose of description and not of limitation, such that the terminology or phraseology of the present specification is to be interpreted by the skilled artisan in light of the teachings and guidance.

The breadth and scope of the present invention should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.

All of the various aspects, embodiments, and options described herein can be combined in any and all variations.

All publications, patents and patent applications mentioned or referenced in this specification are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference.

Claims

A compound of formula I:

or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein:

Z₁ and Z₃ are independently N or CH； provided that Z₁ and Z₃ are not N simultaneously；

Z₂ and Z₄ are independently N or C, in which at least one of Z₂ and Z₄ is N；

R₁ is selected from halo, -CN, C_3-6 cycloalkyl, vinyl, ethynyl, phenyl, and C_1-6 alkyl,

wherein the C_1-6 alkyl is optionally substituted with one or more groups

independently selected from halo, OH, and C_1-6 alkoxyl；

R₂ is C_1-6 alkyl optionally substituted with one or more groups independently selected from C_3-6 cycloalkyl, phenyl and 5-7 membered heteroaryl； or

R₂ is selected from phenyl, C_3-6 cycloalkyl, 4-8 membered heterocycle, and 5-10 membered heteroaryl, each of which is optionally substituted with one or more groups independently selected from halo, -CN, C_1-6 alkyl, -SR^a, -S (O) _nR^b, and C_1-6 alkoxyl, wherein the C_1-6 alkyl and C_1-6 alkoxyl are each optionally substituted by one or more halo；

R₃ and R₄ are each independently selected from H, C_1-6 alkoxyl, and C_1-4 alkyl, wherein the C_1-4 alkyl is optionally substituted by one or more -OH；

R₅ is H or a C_1-4 alkyl；

or R₃, R₅ and the atoms they are attached to form a 4-6 membered mono-or bicyclic, saturated or partially unsaturated heterocyclic ring optionally containing an additional 1-2 heteroatoms selected from N, O and S, which is optionally substituted by one or more groups independently selected from halo and C_1-6 alkyl, wherein the C_1-6 alkyl is optionally substituted by one or more halo；

R^a and R^b are each independently H or a C_1-6 alkyl；

m is 0, 1, or 2；

n is 1 or 2；

W is formula II

wherein:

X₁ is C (O) , X₂ is NR₇, and X₃ is NR₈ or CHR₉；

X₁ is S (O) or S (O) ₂, X₂ is CH₂ or NR₁₀, and X₃ is CH₂； or

X₁ is C (O) , X₂ is CH₂, and X₃ is NH, S or O； or

W is formula II-6-aor formula II-6-b:

wherein R₆ is H, -NH₂, or -NH (C_1-6 alkyl) ； and

R₇, R₈, R₉, and R₁₀ are each independently selected from H, C_1-6 alkyl, and C_3-6 cycloalkyl.
The compound of claim 1, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, m is 1, and R₁ is selected from halo, -CN, methyl, and trifluoromethyl； or m is 0.
The compound of claim 2, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein m is 1 and R₁ is Cl or F.
The compound of any one of claims 1-3, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, R₂ is phenyl or 5-10 membered heteroaryl, each of which is optionally substituted with one or more groups independently selected from halo, -CN, C_1-6 alkyl, -SR^a, -S (O) _nR^b, and C_1-6 alkoxyl, wherein the C_1-6 alkyl and C_1-6 alkoxyl are each optionally substituted by one or more halo.
The compound of claim 4, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, R₂ is phenyl optionally substituted with one or more groups independently selected from halo, -CN, C_1-6 alkyl, -SR^a, -S (O) _nR^b, and C_1-6 alkoxyl, wherein the C_1-6 alkyl and C_1-6 alkoxyl are each optionally substituted by one or more halo； R^a and R^b are each independently C_1-6 alkyl.
The compound of claim 5, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein R₂ is phenyl optionally substituted with one or more halo.
The compound of claim 4, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein R₂ is pyrid-2-yl.
The compound of any one of claims 1-7, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, R₃, R₅ and the atoms they are attached to form
which is optionally substituted by one or more groups independently selected from halo and C_1-6 alkyl, wherein the C_1-6 alkyl is optionally substituted by one or more halo.
The compound of claim 8, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, R₃, R₅ and the atoms they are attached to form
which is optionally substituted by one or more groups independently selected from halo, methyl, ethyl, and trifluoromethyl.
The compound of claim 8, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, R₃, R₅ and the atoms they are attached to form
which is optionally substituted by one or more halo.
The compound of claim 8, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, R₃, R₅ and the atoms they are attached to form
which is optionally substituted by one or more groups independently selected from methyl, and ethyl.
The compound of to any one of claims 1-7, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, R₃ and R₄ are each independently selected from H, C_1-4 alkyl, and C_1-6 alkoxyl； R₅ is H.
The compound of any one of claims 1-12, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, W is selected from II-1 to II-10:
The compound of claim 13, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, W is selected from II-1 to II-4:
The compound of claim 14, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, R₆ is selected from H, -NH₂, and -NH-methyl.
The compound of claim 14, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, R₇, R₈, and R₉ are independently selected from H, methyl, ethyl, and cyclopropyl.
The compound of claim 14, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, formula I is selected from I-1 to I-5:
The compound of claim 17, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, formula I is formula I-1.
The compound of claim 18, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein said formula I-1 is
The compound of claim 13, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, W is selected from I I-5 to I I-7:
The compound of claim 20, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, R₆ is selected from H, -NH₂, and -NH-methyl.
The compound of claim 20, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, R₁₀ is selected from H and t-butyl.
The compound of claim 20, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, formula I is I-1 or I-3:
The compound of claim 23, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein formula I-1 is
The compound of claim 13, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, W is II-8, II-9, or II-10,
The compound of claim 25, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, R₆ is selected from H, and -NH₂.
The compound of claim 25, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein, formula I is
The compound of claim 27, or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salts thereof, wherein said formula I-1 is
The compound of claim 1, wherein m is 0 or 1.
The compound of claim 1, wherein m is 1.
The compound of claim 30, wherein R₁ is selected from halo, -CN, C_3-6 cycloalkyl, phenyl, and C_1-6 alkyl, wherein the C_1-6 alkyl is optionally substituted with one or more groups independently selected from halo, OH, and C_1-6 alkoxyl.
The compound of claim 31, wherein R₁ is selected from F, Cl, Br, Me, CN, CF₃, phenyl, cyclopropyl, methoxylmethyl, and hydroxylmethyl.
The compound of claim 32, wherein R₁ is F or Cl.
The compound of any one of claims 1 and 29-33, wherein R₂ is C_1-6 alkyl optionally substituted with one or more groups independently selected from cyclopropyl and pyridyl； or R₂ is selected from phenyl, cyclopropyl, cyclobutyl, 6-membered heterocycle, and 6-membered heteroaryl, each of which is optionally substituted with one or more groups independently selected from the group consisting of halo, CN, C_1-6 alkyl, -SR^a, -S (O) _nR^b, and C_1-6 alkoxyl, wherein the C_1-6 alkyl and C_1-6 alkoxyl are each optionally substituted by one or more halo； wherein R^a and R^b are each independently a C_1-6 alkyl.
The compound of claim 34, wherein R₂ is phenyl, which is optionally substituted by one or more groups independently selected from F, Cl, CN, CF₃, OMe, OCF₃, OCF₂H, -SMe, -S (O) Me and -S (O) ₂Me.
The compound of claim 35, wherein R₂ is phenyl, which is optionally substituted by one or more groups independently selected from F and Cl.
The compound of claim 34, wherein R₂ is a pyridyl.
The compound of claim 34, wherein R₂ is a 6-membered saturated heterocycle containing one N or O, which is optionally substituted by a C_1-6 alkyl.
The compound of any one of claims 1 and 29-38, wherein R₃ and R₄ are each independently H or a C_1-4 alkyl； R₅ is H or a C_1-4 alkyl；

or R₃, R₅ and the atoms they are attached to form a 4-or 5-membered monocyclic saturated or partially unsaturated heterocyclic ring, which is optionally substituted by one or more groups independently selected from halo and C_1-6 alkyl, wherein the C_1-6 alkyl is optionally substituted by one or more halo.
The compound of claim 39, wherein one of R₃ and R₄ is H, and the other of R₃ and R₄ is a C_1-4 alkyl.
The compound of claim 40, wherein R₅ is H.
The compound of claim 39, wherein R₃, R₅ and the atoms they are attached to form a 4-membered monocyclic saturated ring, which is optionally substituted by one or more groups independently selected from halo and C_1-6 alkyl, wherein the C_1-6 alkyl is optionally substituted by one or more halo.
The compound of claim 39, wherein R₃, R₅ and the atoms they are attached to form a 5-membered monocyclic saturated ring, which is optionally substituted by one or more groups independently selected from halo and C_1-6 alkyl, wherein the C_1-6 alkyl is optionally substituted by one or more halo.
The compound of claims 42 or 43, wherein the 4-or 5-membered monocyclic saturated ring is not substituted.
The compound of claims 42 or 43, wherein the 4-or 5-membered monocyclic saturated ring is substituted by one or more groups independently selected from F, Me, Et, and CF₃.
The compound of claim 45, wherein the 4-or 5-membered monocyclic saturated ring is substituted by one or more groups independently selected from Me and Et.
The compound of any one of claims 1 and 29-46, wherein W is selected from II-1 to II-10,
The compound of claim 47, wherein W is selected from II-1 to II-4,
The compound of claim 47, wherein W is selected from II-5 to II-7,
The compound of claim 47, wherein W is selected from I I-8 to I I-10,
The compound of any one of claims 47-50, wherein R₆ is selected from H, -NH₂, and -NH-methyl.
The compound of claim 51, wherein R₆ is -NH₂.
The compound of any of claims 47-50, 51 and 52, wherein R₇, R₈, and R₉ are independently selected from H, methyl, ethyl, and cyclopropyl.
The compound of claim 53, wherein R₇ is H.
The compound of claim 53, wherein R₇ is Me.
The compound of any one of claims 1 and 29-55, having a structure selected from formula I-1 to I-5:
The compound of claim 56, having a structure selected from formula I-1 and I-3,
The compound of claim 56, having a structure according to formula
The compound of claim 1, selected from compounds numbered 16, 17, 40, 72, 104, and 145.
A compound selected from compounds numbered 1 to 232, or at least one its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or pharmaceutically acceptable salt thereof.
A pharmaceutical composition comprising at least one compound of any of claims 1-60, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.
The pharmaceutical composition of claim 61, comprising at least one compound selected from compounds numbered 16, 17, 40, 72, 104, and 145 or a pharmaceutically acceptable salt thereof.
A method of inhibiting a PI₃K kinase comprising contacting the kinase with an effective amount of at least one compound of any of claims 1-60, or a salt thereof.
A method of treating a disease responsive to inhibition of PI₃K, comprising administrating to a subject in need thereof a therapeutically effective amount of at least one compound of any of claims 1-60, or a pharmaceutically acceptable salt thereof.
The method of claim 64, wherein the disease responsive to inhibition of PI₃K is an immune-based disease or cancer.
The method of claim 65, wherein said immune-based disease is rheumatoid arthritis, COPD, multiple sclerosis, asthma, glomerulonephritis, lupus, psoriasis, or inflammation relating to said disease； wherein said cancer is lymphoma， acute myeloid leukemia, multiple myeloma or chronic lymphocytic leukemia.
The compound of any of claims 1-60, or a pharmaceutically acceptable salt thereof, for use in treatment of a disease responsive to inhibition of PI₃K.
The compound of claim 67, wherein the disease responsive to inhibition of PI₃K is an immune-based disease or cancer.
The compound of claim 68, wherein said immune-based disease is rheumatoid arthritis, COPD, multiple sclerosis, asthma, glomerulonephritis, lupus, psoriasis, or inflammation relating to said disease； wherein said cancer is lymphoma, acute myeloid leukemia, multiple myeloma or chronic lymphocytic leukemia.