WO2014145512A2

WO2014145512A2 - Potent small molecule inhibitors of autophagy, and methods of use thereof

Info

Publication number: WO2014145512A2
Application number: PCT/US2014/030300
Authority: WO
Inventors: Junying Yuan; Nianhe Han; Hua Yi; Yuguang Wang; Song Yang; Jason Christopher WONG
Original assignee: President And Fellows Of Harvard College; F.Hoffmann-Laroche Ltd
Priority date: 2013-03-15
Filing date: 2014-03-17
Publication date: 2014-09-18
Also published as: WO2014145512A3

Abstract

Certain aspects of the invention relate to small molecule autophagy inhibitors, and their use for treatment and prevention of cancers and acute pancreatitis. Medicinal chemistry studies led to small molecular autophagy inhibitors with improved potency and selectivity.

Description

Potent Small Molecule Inhibitors of Autophagy, and Methods of Use Thereof

RELATED APPLICATIONS

This application claims the benefit of priority to United States Provisional Patent Application serial number 61/788,802, filed March 15, 2013; the contents of which are hereby incorporated by reference.

BACKGROUND

Vps34 (vacuolar protein sorting 34), a type III Ptdlns3 kinase (phosphatidylinositol 3 -kinase), was first identified as a regulator of vacuolar hydrolase sorting in yeast (Herman and Emr, 1990). Vps34 specifically phosphorylates the D-3 position on the inositol ring of phosphatidylinositol (Ptdlns) to produce PtdIns3P. PtdIns3P has been implicated in the control of multiple key intracellular membrane trafficking pathways, including endosome to lysosome transport, retrograde endosome to Golgi traffic, multivesicular body formation and autophagy. Two distinct Vps34 phosphatidylinositol 3-kinase complexes function in autophagy and carboxypeptidase Y sorting in Saccharomyces cerevisiae. PtdIns3P is required for the initiation of autophagy, an evolutionarily conserved catabolic mechanism involved in the turnover of intracellular organelles and large protein complexes.

Vps34 is present in two complexes in yeast: complex I (Vps34, Vpsl5, Vps30/Atg6, and Atgl4) involved in autophagy, and complex II (Vps34, Vpsl5, Vps30/Atg6, and Vps38) in the vacuolar protein sorting pathway. In mammalian cells, Vps34 is found in at least two protein complexes, Vps34 complex I and Vps34 complex II, that may function similarly to their homologous complexes in yeast. The two mammalian Vps34 complexes share the core components of Vps34, Beclinl and pi 50, which are homologous to yeast Vps34, Vps30/Atg6 and Vpsl5, respectively. In addition, the complex I contains Atgl4L, the mammalian orthologue of yeast Atgl4, which localizes to the isolation membrane/phagophore during starvation and is essential for autophagosome formation; while the complex II contains UVRAG, a homologue of Vps38 in yeast, which primarily localizes to late endosomes. Interestingly, the stabilities of different components of Vps34 complexes are co-dependent upon each other as knockdown of one component often reduces the levels of others in the complexes. However, very little is known about the mechanisms that regulate the stability of Vps34 complexes which may play an important role in regulating multiple vesicular trafficking pathways.

Autophagy is a catabolic process mediating the turnover of intracellular constituents in a lysosome-dependent manner. Autophagy is initiated by the formation of an isolation membrane, which expands to engulf a portion of cytoplasm, including large protein complexes and defective organelles, by forming a double membrane vesicle, termed autophagosome. The contents of an autophagosome are degraded by lysosomal hydrolases after its fusion with a lysosome to form an autolysosome. Autophagy has been studied extensively in unicellular eukaryotes as a strategy to survive starvation conditions, as products of autophagic degradation such as free amino acids, fatty acids and nucleotides, can be used by the cell as building blocks or a source of energy in order to help survive under nutrient limiting conditions.

The core molecular machinery of autophagy is controlled by the protein products encoded by a group of ATG genes evolutionarily conserved from yeast to mammals. Nucleation of autophagic vesicles requires PtdIns3P, the product of type III PI3 kinase complex including Beclin 1 (mammalian homolog of yeast Atg6) and Vps34, as well as two ubiquitin-like molecules, Atgl2 and LC3 (homolog of Atg8), which function sequentially in mediating the formation of autophagosomes. In the first ubiquitination-like reaction, Atgl2 is conjugated to Atg5 and forms a large multimeric protein complex, which plays a key role in determining the nucleation of autophagosome. In the second reaction, LC3 is conjugated to phosphatidyl-ethanolamine, resulting in membrane translocation important for the elongation and closure of autophagosome.

In metazoans, autophagy functions as an essential intracellular catabolic mechanism involved in cellular homeostasis by mediating the turnover of malfunctioning, aged or damaged proteins and organelles. Down-regulation of autophagy contributes to neurodegeneration by increasing the accumulation of misfolded proteins. Autophagy can also be activated in response to many forms of cellular stress beyond nutrient starvation, including DNA damage, ER stress and invasion by intracellular pathogens, and has been shown to participate in both innate and acquired immunity as well as in tumor suppression. Mechanisms that regulate autophagy in mammalian cells are just beginning to be explored.

Autophagy plays an important role in regulating cellular homeostasis and contributes to cell survival, growth, differentiation and host defense responses. Dysregulation of autophagy has been implicated in multiple human diseases including cancer, neurodegeneration, inflammatory diseases and infectious diseases. Most of the currently knowledge on autophagy were derived from elegant genetic studies in yeast which led to the identification of autophagy "Atg" genes . Recent studies have demonstrated the evolutionary conservation of the core autophagy genes from yeast to mammal; however, the mechanism and regulation of mammalian autophagy have shown significant increases in the complexity which very little is known.

Autophagy has been proposed to play complex roles in the development and treatment of cancers. Activation of autophagy may promote tumor cell survival under metabolic stress and function as a tumor suppression mechanism by preventing necrotic cell death and subsequent inflammation which favors tumor growth. On the other hand, inhibition of autophagy may lead to genome instability through unknown mechanisms which might explain the increased frequency of beclin 1 heterozygosity in multiple lines of cancers and decreased expression of autophagy-related proteins in malignant epithelial ovarian cancer. Thus, chronic suppression of autophagy may stimulate tumorigenesis.

The proposed role of autophagy in anticancer therapy is opposite to that during tumorigenesis. Once a tumor is formed, acute inhibition of autophagy might be beneficial for the therapeutic goal by promoting radiosensitization and chemosensitization. In an animal model of cancer therapy, inhibition of therapy-induced autophagy either with shRNA against a key autophagy gene ATG5 or with anti-malarial drug chloroquine enhanced cell death and tumor regression of Myc-driven tumors in which either activated p53 or alkylating chemotherapy was used to drive tumor cell death. Chloroquine causes a dose-dependent accumulation of large autophagic vesicles and enhances alkylating therapy- induced cell death to a similar degree as knockdown of ATG5. In another example, resistance to TRAIL was found to be reversed by a common approach of targeting specific components of autophagic process, such as Beclinl or Vps34, for inhibition. In the case of chronic myelogenous leukemia (CML), inhibition of autophagy by chloroquine markedly enhanced death of a CML cell line, K562, induced by imatinib. Furthermore, imatinib- resistant cell lines, BaF3/T315I and BaF3/E255K, can be induced to die by co-treatment with imatinib and chloroquine. Thus, inhibition of autophagy sensitizes tumor cells to imatinib-induced cell death. The block of autophagy has been proposed to be a new strategy for the treatment of CML. These studies suggest that autophagy can promote resistance to DNA-damaging therapy.

In addition, autophagy has also been shown to play an important role in mediating cellular damage induced by acute pancreatitis. Autodigestion of the pancreas by its own prematurely activated digestive proteases is thought to be an important event in the onset of acute pancreatitis. A conditional knockout mouse that lacks the autophagy-related (Atg) gene Atg5 in the pancreatic acinar cells has shown significantly reduced severity of acute pancreatitis induced by cerulein. Thus autophagy exerts a detrimental effect in pancreatic acinar cells by activation of trypsinogen to trypsin. Inhibitors of autophagy may therefore provide important new therapeutics for acute pancreatitis.

Further, small molecule inhibitors are important tools in exploring the cellular mechanisms in mammalian cells. However, the only available small molecule inhibitor of autophagy is 3-methyladenine (3-MA), which has a working concentration of about 10 mM and is highly non-specific. Therefore, there is a need for small molecule autophagy inhibitors, and in particular, small molecule autophagy inhibitors for the treatment and prevention of cancers and acute pancreatitis.

SUMMARY

In certain embodiments, the invention relates to a compound represented by Formula IV, Formula V, or Formula VI:

Formula VI

wherein, independently for each occurrence,

Y is F, CN, NO2, S0₂R, SO2NR2, NRSO2R, I, CH₃, CI, CF₃, or CONR₂;

R² is halo, aryloxy, arylamino, N0₂, OH, NRCOR, C0₂R, NRS0₂R, S0₂NR₂, CONR₂, S0₂R, CN, NH₂, -C(OH)(CF₃)₂, NRC(0)NR₂, -OCH₂CH₂-(N-morpholinyl),

_s ^ό _{s s or s or an}y two adjacent R², taken together, form a five- or six-membered amide ring;

R² may be present on the saturated ring or the unsaturated ring;

m is 0, 1, or 2; and

R is alkyl, cycloalkyl, H, hydroxyalkyl, aminoalkyl, alkoxyalkyl, fiuoroalkylalkyl, heterocyclyl, or cyanoalkyl.

In certain embodiments, the invention relates to a compound represented by Formula VII

Formula VII

wherein, independently for each occurrence,

Y is F, CN, N0₂, S0₂R, S0₂NR₂, NRS0₂R, I, CH₃, CI, CF₃, or CONR₂;

q is 1 or 2;

n is 0, 1, or 2;

R¹ is NRCOR, C0₂R, NRS0₂R, S0₂ -C(OH)(CF₃)₂,

NRC(0)NR₂, -OCH₂CH₂-(N-morpholinyl),

, or any two adjacent R¹, taken together, form a five- or six-membered amide ring; or, when q > 2, one instance of R¹ may be halo; and

R is alkyl, cycloalkyl, H, hydroxyalkyl, aminoalkyl, alkoxyalkyl, fiuoroalkylalkyl, heterocyclyl, or cyanoalkyl. In certain embodiments, the invention relates to a compound represented by Formula VIII

Formula VIII

wherein, independently for each occurrence,

Y is F, CN, NO2, S0₂R, SO2NR2, NRSO2R, I, CH₃, CI, CF₃, or CONR₂;

A is pyrrolyl, pyrazinyl, or indolyl, any of which may be substituted with one or more R²;

R² is halo, aryloxy, arylamino, N0₂, OH, NRCOR, C0₂R, NRS0₂R, S0₂NR₂,

H₂, -C(OH)(CF₃)₂, NRC(0)NR₂, -OCH₂CH₂-(N-morpholinyl),

or any two adjacent R², taken together, form a five- or six-membered amide ring;

n is 0, 1, or 2; and

R is alkyl, cycloalkyl, H, hydroxyalkyl, aminoalkyl, alkoxyalkyl, fluoroalkylalkyl, heterocyclyl, or cyanoalkyl.

In certain embodiments, the invention relates to a compound represented by Formula IX

Formula IX

wherein, independently for each occurrence,

Y is F, CN, NO2, S0₂R, SO2NR2, NRSO2R, I, CH₃, CI, CF₃, or CONR₂;

R² is halo, aryloxy, arylamino, N0₂, OH, NRCOR, C0₂R, NRS0₂R, S0₂NR₂, CONR2, S0₂R, CN, NH₂, -C(OH)(CF₃)₂, NRC(0)NR₂, -OCH₂CH₂-(N-morpholinyl),

, or any two adjacent R², taken together, form a five- or six-membered amide ring;

m is 0, 1, or 2;

n is 0, 1, or 2; and

In certain embodiments, the invention relates to a compound of Formula I, Formula II, or Formula III, as depicted in Figure 2, wherein Ri is Y as defined above; R₂- R₇ is H or R² as defined above; n is 0, 1, or 2; and the 2-substituent in Figure 2 (R) is H, halo, CN, N0₂, fluoroalkyl, or alkyl.

In certain embodiments, the invention relates to a compound selected from the group consisting of the compounds listed in any of the Figures. In certain embodiments, the invention relates to a compound selected from the group consisting of the compounds listed in Figure 1.

In certain embodiments, the invention relates to a method for inhibiting autophagy in a subject for whom inhibition of autophagy is beneficial, comprising administering to the subject a therapeutically effective amount of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby inhibiting autophagy activity in the subject.

In certain embodiments, the invention relates to a method of treating or preventing cancer, comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby treating or preventing cancer.

In certain embodiments, the invention relates to a method of treating or preventing cancer, comprising the step of co-administering to a subject in need thereof (i) a therapeutically effective amount of a compound of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, and (ii) a therapeutically effective amount of a second active agent, thereby treating or preventing cancer. In certain embodiments, the invention relates to a method of treating or preventing acute pancreatitis, comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby treating or preventing pancreatitis.

In certain embodiments, the invention relates to a method of treating or preventing a disease caused by an intracellular pathogen, comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby treating or preventing the disease caused by the intracellular pathogen.

In certain embodiments, the invention relates to a method of treating or preventing a lysosomal storage disorder, comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby treating or preventing the lysosomal storage disorder.

In certain embodiments, the invention relates to a method of inhibiting autophagy in a cell in need thereof, comprising the step of contacting the cell with a therapeutically effective amount of a compound of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby inhibiting autophagy in the cell.

In certain embodiments, the invention relates to a method of inactivating a deubiquitinating protease complex, comprising the step of contacting the deubiquitinating protease complex with a compound of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, wherein the deubiquitinating protease complex comprises USP3 and USP10.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1 tabulates various compounds of the invention.

Figure 2 depicts three generic structures of compounds of the invention. Figure 3 depicts a schematic representation of a general approach to the synthesis of compounds of the invention.

Figure 4 depicts a schematic representation of a synthesis of compound A02-172.

Figure 5 depicts a schematic representation of a synthesis of compound A02-171.

Figure 6 depicts a schematic representation of a synthesis of compound A23-001.

Figure 7 depicts three generic structures of compounds of the invention.

Figure 8 tabulates a series of tetrahydronaphthalene derivatives as autophagy inhibitors.

Figure 9 tabulates a selective group of indene-containing compounds as autophagy inhibitors.

Figure 10 tabulates a selective group of tetrahydrobenzothiazole-containing compounds as autophagy inhibitors.

Figure 11 tabulates the combination effect of compounds of the invention with AC220 in eight p53 mutant breast and ovarian cancer cell lines.

Figure 12 tabulates the combination effect of compounds of the invention with AC220 in ES-2 cell line.

Figure 13 tabulates the combination effect of compounds of the invention with AC220 in SUM 159 cell line.

Figure 14 tabulates the compound profile in breast and ovarian cells lines in glucose free conditions.

Figures 15-69 depict schematic representations of syntheses of compounds of the invention.

Figure 70 tabulates the potency of autophagy inhibition of various compounds of the invention.

Figure 71 depicts some compounds of the invention used in studies of structure- activity relationships (SAR).

Figure 72 depicts compounds of the invention.

Figure 73 depicts the results from the combination study of A31-001 A with AC220 on ES-2 cells.

Figure 74 depicts the results from the combination study of A31-00 IB with AC220 on ES-2 cells.

Figure 75 tabulates the structures and various biological properties of some compounds of the invention. DETAILED DESCRIPTION

Overview

Autophagy, a cellular catabolic process, plays an important role in promoting cell survival under metabolic stress condition by mediating lysosomal-dependent turnover of intracellular constituents for recycling. Certain aspects of the invention relate to small molecule autophagy inhibitors, and their use for treatment and prevention of cancers and acute pancreatitis.

Definitions

For convenience, certain terms employed in the specification, examples, and appended claims are collected here. All definitions, as defined and used herein, supersede dictionary definitions, definitions in documents incorporated by reference, and/or ordinary meanings of the defined terms.

The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

The phrase "and/or," as used herein in the specification and in the claims, should be understood to mean "either or both" of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with "and/or" should be construed in the same fashion, i.e., "one or more" of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the "and/or" clause, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, a reference to "A and/or B", when used in conjunction with open-ended language such as "comprising" can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.

As used herein in the specification and in the claims, "or" should be understood to have the same meaning as "and/or" as defined above. For example, when separating items in a list, "or" or "and/or" shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as "only one of or "exactly one of," or, when used in the claims, "consisting of," will refer to the inclusion of exactly one element of a number or list of elements. In general, the term "or" as used herein shall only be interpreted as indicating exclusive alternatives (i.e., "one or the other but not both") when preceded by terms of exclusivity, such as "either," "one of," "only one of," or "exactly one of." "Consisting essentially of," when used in the claims, shall have its ordinary meaning as used in the field of patent law.

As used herein in the specification and in the claims, the phrase "at least one," in reference to a list of one or more elements, should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase "at least one" refers, whether related or unrelated to those elements specifically identified. Thus, as a non- limiting example, "at least one of A and B" (or, equivalently, "at least one of A or B," or, equivalently "at least one of A and/or B") can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc.

It should also be understood that, unless clearly indicated to the contrary, in any methods claimed herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited.

In the claims, as well as in the specification above, all transitional phrases such as "comprising," "including," "carrying," "having," "containing," "involving," "holding," "composed of," and the like are to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases "consisting of and "consisting essentially of shall be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03.

The definition of each expression, e.g., alkyl, m, n, and the like, when it occurs more than once in any structure, is intended to be independent of its definition elsewhere in the same structure. It will be understood that "substitution" or "substituted with" includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, or other reaction.

The term "substituted" is also contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, for example, those described herein below. The permissible substituents may be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. This invention is not intended to be limited in any manner by the permissible substituents of organic compounds. When "one or more" substituents are indicated, there may be, for example, 1, 2, 3, 4 or 5 substituents.

The term "lower" when appended to any of the groups listed below indicates that the group contains less than seven carbons (i.e., six carbons or less). For example "lower alkyl" refers to an alkyl group containing 1-6 carbons.

For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover.

The term "alkyl" means an aliphatic or cyclic hydrocarbon radical containing from 1 to 20, 1 to 15, or 1 to 10 carbon atoms. Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n- pentyl, isopentyl, neopentyl, n-hexyl, 2-methylcyclopentyl, and 1-cyclohexylethyl. The term "fluoroalkyl" means an alkyl wherein one or more hydrogens are replaced with fluorines.

The term "alkoxy" means an alkyl group bound to the parent moiety through an oxygen. The term "fluoroalkoxy" means a fluoroalkyl group bound to the parent moiety through an oxygen. Selected Autophagy Inhibitors

Formula IV

Formula VI

wherein, independently for each occurrence,

_{s or atl}y wo adjacent R², taken together, form a five- or six-membered amide ring;

R² may be present on the saturated ring or the unsaturated ring;

m is 0, 1, or 2; and

R is alkyl, cycloalkyl, H, hydroxyalkyl, aminoalkyl, alkoxyalkyl, fluoroalkylalkyl, heterocyclyl, or cyanoalkyl. In certain embodiments, the invention relates to a compound represented by Formula VII

Formula VII

wherein, independently for each occurrence,

Y is F, CN, NO2, S0₂R, SO2NR2, NRSO2R, I, CH₃, CI, CF₃, or CONR₂;

q is 1 or 2;

n is 0, 1, or 2;

R¹ is NRCOR, C0₂R, NRSO2R, SO2NR2, CONR2, S0₂R, CN, NH₂, -C(OH)(CF₃)₂,

NRC(0)NR₂, -OCH₂CH₂-(N-morpholinyl),

, - ^, , , or , or any two adjacent R¹, taken together, form a five- or six-membered amide ring; or, when q > 2, one instance of R¹ may be halo; and

In certain embodiments, the invention relates to a compound represented by Formula VIII

Formula VIII

wherein, independently for each occurrence,

Y is F, CN, NO2, S0₂R, SO2NR2, NRS0₂R, I, CH₃, CI, CF₃, or CONR₂;

n is 0, 1, or 2; and

Formula IX

wherein, independently for each occurrence,

taken together, form a five- or six-membered amide ring;

m is 0, 1, or 2;

n is 0, 1, or 2; and

In certain embodiments, the invention relates to a compound of Formula I, Formula II, or Formula III, as depicted in Figure 2, wherein Ri is Y as defined above; R₂- R₇ is H or R² as defined above; n is 0, 1, or 2; and the 2-substituent in Figure 2 (R) is H, halo, CN, N0₂, fluoroalkyl, or alkyl. Please note that the 2-substituent in Figure 2 is NOT the same as R as defined in Formulas I-IX above.

In certain embodiments, the invention relates to a compound selected from the group consisting of the compounds listed in Figure 1.

In certain embodiments, the invention relates to any of the aforementioned compounds and attendant definitions, wherein the compound is an autophagy inhibitor; and the EC₅₀ of the autophagy inhibitor is less than about 100 nM.

In certain embodiments, the invention relates to any one of the aforementioned compounds, wherein the compound inhibits autophagy with an IC₅₀ of less than about 10 μΜ. In certain embodiments, the invention relates to any one of the aforementioned compounds, wherein the compound inhibits autophagy with an IC₅₀ of less than about 5 μΜ. In certain embodiments, the invention relates to any one of the aforementioned compounds, wherein the compound inhibits autophagy with an IC₅₀ of less than about 1 μΜ. In certain embodiments, the invention relates to any one of the aforementioned compounds, wherein the compound inhibits autophagy with an IC₅₀ of less than about 750 nM. In certain embodiments, the invention relates to any one of the aforementioned compounds, wherein the compound inhibits autophagy with an IC₅₀ of less than about 500 nM. In certain embodiments, the invention relates to any one of the aforementioned compounds, wherein the compound inhibits autophagy with an IC₅₀ of less than about 250 nM. In certain embodiments, the invention relates to any one of the aforementioned compounds, wherein the compound inhibits autophagy with an IC₅₀ of less than about 100 nM.

Certain compounds of the invention which have acidic substituents may exist as salts with pharmaceutically acceptable bases. The present invention includes such salts. Examples of such salts include sodium salts, potassium salts, lysine salts and arginine salts. These salts may be prepared by methods known to those skilled in the art.

Certain compounds of the invention and their salts may exist in more than one crystal form and the present invention includes each crystal form and mixtures thereof.

Certain compounds of the invention and their salts may also exist in the form of solvates, for example hydrates, and the present invention includes each solvate and mixtures thereof.

Certain compounds of the invention may contain one or more chiral centers, and exist in different optically active forms. When compounds of the invention contain one chiral center, the compounds exist in two enantiomeric forms and the present invention includes both enantiomers and mixtures of enantiomers, such as racemic mixtures. The enantiomers may be resolved by methods known to those skilled in the art, for example by formation of diastereoisomeric salts which may be separated, for example, by crystallization; formation of diastereoisomeric derivatives or complexes which may be separated, for example, by crystallization, gas-liquid or liquid chromatography; selective reaction of one enantiomer with an enantiomer-specific reagent, for example enzymatic esterification; or gas-liquid or liquid chromatography in a chiral environment, for example on a chiral support for example silica with a bound chiral ligand or in the presence of a chiral solvent. It will be appreciated that where the desired enantiomer is converted into another chemical entity by one of the separation procedures described above, a further step may be used to liberate the desired enantiomeric form. Alternatively, specific enantiomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer into the other by asymmetric transformation.

When a compound of the invention contains more than one chiral center, it may exist in diastereoisomeric forms. The diastereoisomeric compounds may be separated by methods known to those skilled in the art, for example chromatography or crystallization and the individual enantiomers may be separated as described above. The present invention includes each diastereoisomer of compounds of the invention and mixtures thereof.

Certain compounds of the invention may exist in different tautomeric forms or as different geometric isomers, and the present invention includes each tautomer and/or geometric isomer of compounds of the invention and mixtures thereof.

Certain compounds of the invention may exist in different stable conformational forms which may be separable. Torsional asymmetry due to restricted rotation about an asymmetric single bond, for example because of steric hindrance or ring strain, may permit separation of different conformers. The present invention includes each conformational isomer of compounds of the invention and mixtures thereof.

Certain compounds of the invention may exist in zwitterionic form and the present invention includes each zwitterionic form of compounds of the invention and mixtures thereof.

As used herein the term "pro-drug" refers to an agent which is converted into the parent drug in vivo by some physiological chemical process (e.g., a prodrug on being brought to the physiological pH is converted to the desired drug form). Pro-drugs are often useful because, in some situations, they may be easier to administer than the parent drug. They may, for instance, be bioavailable by oral administration whereas the parent drug is not. The prodrug may also have improved solubility in pharmacological compositions over the parent drug. An example, without limitation, of a pro-drug would be a compound of the present invention wherein it is administered as an ester (the "pro-drug") to facilitate transmittal across a cell membrane where water solubility is not beneficial, but then it is metabolically hydrolyzed to the carboxylic acid once inside the cell where water solubility is beneficial. Pro-drugs have many useful properties. For example, a pro-drug may be more water soluble than the ultimate drug, thereby facilitating intravenous administration of the drug. A pro-drug may also have a higher level of oral bioavailability than the ultimate drug. After administration, the prodrug is enzymatically or chemically cleaved to deliver the ultimate drug in the blood or tissue.

Exemplary pro-drugs release an amine of a compound of the invention wherein the free hydrogen of an amine is replaced by (Ci-C₆)alkanoyloxymethyl, l-((Ci- C₆)alkanoyloxy)ethyl, 1 -methyl- 1 -((C i-Ce)alkanoyloxy)ethyl, (C i -

C₆)alkoxycarbonyloxymethyl, N-(Ci-C₆)alkoxycarbonylaminomethyl, succinoyl, (Ci- C₆)alkanoyl, a-amino(Ci-C4)alkanoyl, arylactyl and a-aminoacyl, or a-aminoacyl-a- aminoacyl wherein said a-aminoacyl moieties are independently any of the naturally occurring L-amino acids found in proteins, -P(0)(OH)₂, -P(0)(0(Ci-C₆)alkyl)₂ or glycosyl (the radical resulting from detachment of the hydroxyl of the hemiacetal of a carbohydrate).

In certain embodiments, the invention relates to any one of the aforementioned compounds, wherein the compound is not disclosed in US Patent Application Publication No. 2012/0258975.

In certain embodiments, the invention relates to any one of the aforementioned compounds, wherein the compound is not A02-010, A02-011, A02-013, A02-018, A02- 034, A02-035, A02-036, A02-065, A02-103, A02-105, A02-122, A02-125, A02-170, A02- 181, A02-182, A22-009, A22-016, A22-017, A22-019, A22-022, A22-023, A22-024, A22- 025, A22-028, A22-031, A22-035, A22-040, A22-044, A22-045, A22-064, A22-083, A22- 096, A22-097, A22-100, A22-101, A22-102, A22-103, A22-113, A22-114, A22-118, A22- 119, A22-127, A22-128, A22-129, A22-130, A22-139, A22-140, A22-143, A22-144, A23- 005, A23-007, A23-008, A24-001, A24-002, A24-003, A24-004, A24-005, A24-008, A24- 009, A24-010, A25-001, A25-002, A26-001, A26-002, A26-003, A26-004, A26-005, A27- 002, A27-004, A27-005, A27-006, A27-007, A28-003, or A28-004,

Pharmaceutical Compositions

One or more compounds of this invention can be administered to a human patient by themselves or in pharmaceutical compositions where they are mixed with biologically suitable carriers or excipient(s) at doses to treat or ameliorate a disease or condition as described herein. Mixtures of these compounds can also be administered to the patient as a simple mixture or in suitable formulated pharmaceutical compositions. For example, one aspect of the invention relates to pharmaceutical composition comprising a therapeutically effective dose of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof; and a pharmaceutically acceptable diluent or carrier.

As used herein, a therapeutically effective dose refers to that amount of the compound or compounds sufficient to result in the prevention or attenuation of a disease or condition as described herein. Techniques for formulation and administration of the compounds of the instant application may be found in references well known to one of ordinary skill in the art, such as "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, PA, latest edition.

Suitable routes of administration may, for example, include oral, eyedrop, rectal, transmucosal, topical, or intestinal administration; parenteral delivery, including intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, or intraocular injections.

Alternatively, one may administer the compound in a local rather than a systemic manner, for example, via injection of the compound directly into an edematous site, often in a depot or sustained release formulation.

Furthermore, one may administer the drug in a targeted drug delivery system, for example, in a liposome coated with endothelial cell-specific antibody.

The pharmaceutical compositions of the present invention may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes.

Pharmaceutical compositions for use in accordance with the present invention thus may be formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen.

For injection, the agents of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks' solution, Ringer's solution, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

For oral administration, the compounds can be formulated readily by combining the active compounds with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated. Pharmaceutical preparations for oral use can be obtained by combining the active compound with a solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.

Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration.

For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

For administration by inhalation, the compounds for use according to the present invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebuliser, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of e.g., gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

The compounds can be formulated for parenteral administration by injection, e.g., bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.

Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.

Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen- free water, before use.

The compounds may also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides. In addition to the formulations described previously, the compounds may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly or by intramuscular injection). Thus, for example, the compounds may be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

Alternatively, other delivery systems for hydrophobic pharmaceutical compounds may be employed. Liposomes and emulsions are well known examples of delivery vehicles or carriers for hydrophobic drugs. Certain organic solvents such as dimethysulfoxide also may be employed, although usually at the cost of greater toxicity. Additionally, the compounds may be delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained-release materials have been established and are well known by those skilled in the art. Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic reagent, additional strategies for protein stabilization may be employed.

The pharmaceutical compositions also may comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include but are not limited to calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.

Many of the compounds of the invention may be provided as salts with pharmaceutically compatible counterions (i.e., pharmaceutically acceptable salts). A "pharmaceutically acceptable salt" means any non-toxic salt that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound or a prodrug of a compound of this invention. A "pharmaceutically acceptable counterion" is an ionic portion of a salt that is not toxic when released from the salt upon administration to a recipient. Pharmaceutically compatible salts may be formed with many acids, including but not limited to hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, etc. Salts tend to be more soluble in aqueous or other protonic solvents than are the corresponding free base forms.

Acids commonly employed to form pharmaceutically acceptable salts include inorganic acids such as hydrogen bisulfide, hydrochloric, hydrobromic, hydroiodic, sulfuric and phosphoric acid, as well as organic acids such as para-toluenesulfonic, salicylic, tartaric, bitartaric, ascorbic, maleic, besylic, fumaric, gluconic, glucuronic, formic, glutamic, methanesulfonic, ethanesulfonic, benzenesulfonic, lactic, oxalic, para- bromophenylsulfonic, carbonic, succinic, citric, benzoic and acetic acid, and related inorganic and organic acids. Such pharmaceutically acceptable salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4- dioate, hexyne-l,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, sulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, .beta.-hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene- 1 -sulfonate, naphthalene -2-sulfonate, mandelate and the like salts. Preferred pharmaceutically acceptable acid addition salts include those formed with mineral acids such as hydrochloric acid and hydrobromic acid, and especially those formed with organic acids such as maleic acid.

Suitable bases for forming pharmaceutically acceptable salts with acidic functional groups include, but are not limited to, hydroxides of alkali metals such as sodium, potassium, and lithium; hydroxides of alkaline earth metal such as calcium and magnesium; hydroxides of other metals, such as aluminum and zinc; ammonia, and organic amines, such as unsubstituted or hydroxy-substituted mono-, di-, or trialkylamines; dicyclohexylamine; tributyl amine; pyridine; N-methyl-N-ethylamine; diethylamine; triethylamine; mono-, bis-, or tris-(2-hydroxy-lower alkyl amines), such as mono-, bis-, or tris-(2-hydroxyethyl)amine, 2-hydroxy-tert-butylamine, or tris-(hydroxymethyl)methylamine, N,N-di alkyl-N-(hydroxy alkyl)-amines, such as N,N-dimethyl-N-(2-hydroxyethyl)amine, or tri-(2- hydroxyethyl)amine; N-methyl-D-glucamine; and amino acids such as arginine, lysine, and the like.

Pharmaceutical compositions suitable for use in the present invention include compositions wherein the active ingredients are contained in an effective amount to achieve its intended purpose. More specifically, a therapeutically effective amount means an amount effective to prevent development of or to alleviate the existing symptoms of the subject being treated. Determination of the effective amounts is well within the capability of those skilled in the art.

Selected Methods of Use

One aspect the invention provides a method for inhibiting autophagy in a subject for whom inhibition of autophagy is beneficial, comprising administering to the subject an effective amount of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby inhibiting autophagy activity in the subject. In certain embodiments, the subject is a human.

The term "subject" for purposes of treatment includes any human or animal subject who has been diagnosed with, has symptoms of, or is at risk of developing a disorder wherein inhibition of autophagy would be beneficial. For methods of prevention the subject is any human or animal subject. To illustrate, for purposes of prevention, a subject may be a human subject who is at risk of or is genetically predisposed to obtaining a disorder characterized by unwanted, rapid cell proliferation, such as cancer. The subject may be at risk due to exposure to carcinogenic agents, being genetically predisposed to disorders characterized by unwanted, rapid cell proliferation, and so on. Besides being useful for human treatment, the compounds described herein are also useful for veterinary treatment of mammals, including companion animals and farm animals, such as, but not limited to dogs, cats, horses, cows, sheep, and pigs.

One aspect of the invention relates to a method of treating or preventing cancer, comprising the step of administering to a subject in need thereof a therapeutically effective amount of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby treating or preventing cancer.

One aspect of the invention relates to a method of treating or preventing cancer, comprising the step of co-administering to a subject in need thereof (i) a therapeutically effective amount of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, and (ii) a therapeutically effective amount of a second active agent, thereby treating or preventing cancer.

The term "treating" as used herein, encompasses the administration and/or application of one or more compounds described herein, to a subject, for the purpose of providing prevention of or management of, and/or remedy for a condition. "Treatment" for the purposes of this disclosure, may, but does not have to, provide a cure; rather, "treatment" may be in the form of management of the condition. When the compounds described herein are used to treat unwanted proliferating cells, including cancers, "treatment" includes partial or total destruction of the undesirable proliferating cells with minimal destructive effects on normal cells. A desired mechanism of treatment of unwanted rapidly proliferating cells, including cancer cells, at the cellular level is apoptosis.

The term "preventing" as used herein includes either preventing or slowing the onset of a clinically evident unwanted cell proliferation altogether or preventing or slowing the onset of a preclinically evident stage of unwanted rapid cell proliferation in individuals at risk. Also intended to be encompassed by this definition is the prevention or slowing of metastasis of malignant cells or to arrest or reverse the progression of malignant cells. This includes prophylactic treatment of those at risk of developing precancers and cancers. Also encompassed by this definition is the prevention or slowing of restenosis in subjects that have undergone angioplasty or a stent procedure.

Suppression of autophagy has been proposed to be a new anticancer therapy by promoting radiosensitization and chemosensitization. In an animal model of cancer therapy, inhibition of therapy-induced autophagy either with shR A against a key autophagy gene ATG5 or with anti-malarial drug chloroquine enhanced cell death and tumor regression of Myc-driven tumors in which either activated p53 or alkylating chemotherapy was used to drive tumor cell death (Amaravadi, R.K., et al., Autophagy inhibition enhances therapy- induced apoptosis in a Myc-induced model of lymphoma. J Clin Invest, 2007. 117(2): p. 326-36). Chloroquine causes a dose-dependent accumulation of large autophagic vesicles and enhances alkylating therapy-induced cell death to a similar degree as knockdown of ATG5. In the case of chronic myelogenous leukemia (CML), chloroquine markedly enhanced death of a CML cell line, K562, induced by imatinib. Furthermore, imatinib- resistant cell lines, BaF3/T315I and BaF3/E255K, can be induced to die by co-treatment with imatinib and chloroquine. These studies suggest that inhibiting autophagy may potentiate conventional chemotherapy.

The National Cancer Institute alphabetical list of cancer includes: Acute Lymphoblastic Leukemia, Adult; Acute Lymphoblastic Leukemia, Childhood; Acute Myeloid Leukemia, Adult; Adrenocortical Carcinoma; Adrenocortical Carcinoma, Childhood; AIDS-Related Lymphoma; AIDS-Related Malignancies; Anal Cancer; Astrocytoma, Childhood Cerebellar; Astrocytoma, Childhood Cerebral; Bile Duct Cancer, Extrahepatic; Bladder Cancer; Bladder Cancer, Childhood; Bone Cancer, Osteosarcoma/Malignant Fibrous Histiocytoma; Brain Stem Glioma, Childhood; Brain Tumor, Adult; Brain Tumor, Brain Stem Glioma, Childhood; Brain Tumor, Cerebellar Astrocytoma, Childhood; Brain Tumor, Cerebral Astrocytoma/Malignant Glioma, Childhood; Brain Tumor, Ependymoma, Childhood; Brain Tumor, Medulloblastoma, Childhood; Brain Tumor, Supratentorial Primitive Neuroectodermal Tumors, Childhood; Brain Tumor, Visual Pathway and Hypothalamic Glioma, Childhood; Brain Tumor, Childhood (Other); Breast Cancer; Breast Cancer and Pregnancy; Breast Cancer, Childhood; Breast Cancer, Male; Bronchial Adenomas/Carcinoids, Childhood; Carcinoid Tumor, Childhood; Carcinoid Tumor, Gastrointestinal; Carcinoma, Adrenocortical; Carcinoma, Islet Cell; Carcinoma of Unknown Primary; Central Nervous System Lymphoma, Primary; Cerebellar Astrocytoma, Childhood; Cerebral Astrocytoma/Malignant Glioma, Childhood; Cervical Cancer; Childhood Cancers; Chronic Lymphocytic Leukemia; Chronic Myelogenous Leukemia; Chronic Myeloproliferative Disorders; Clear Cell Sarcoma of Tendon Sheaths; Colon Cancer; Colorectal Cancer, Childhood; Cutaneous T-Cell Lymphoma; Endometrial Cancer; Ependymoma, Childhood; Epithelial Cancer, Ovarian; Esophageal Cancer; Esophageal Cancer, Childhood; Ewing's Family of Tumors; Extracranial Germ Cell Tumor, Childhood; Extragonadal Germ Cell Tumor; Extrahepatic Bile Duct Cancer; Eye Cancer, Intraocular Melanoma; Eye Cancer, Retinoblastoma; Gallbladder Cancer; Gastric (Stomach) Cancer; Gastric (Stomach) Cancer, Childhood; Gastrointestinal Carcinoid Tumor; Germ Cell Tumor, Extracranial, Childhood; Germ Cell Tumor, Extragonadal; Germ Cell Tumor, Ovarian; Gestational Trophoblastic Tumor; Glioma, Childhood Brain Stem; Glioma, Childhood Visual Pathway and Hypothalamic; Hairy Cell Leukemia; Head and Neck Cancer; Hepatocellular (Liver) Cancer, Adult (Primary); Hepatocellular (Liver) Cancer, Childhood (Primary); Hodgkin's Lymphoma, Adult; Hodgkin's Lymphoma, Childhood; Hodgkin's Lymphoma During Pregnancy; Hypopharyngeal Cancer; Hypothalamic and Visual Pathway Glioma, Childhood; Intraocular Melanoma; Islet Cell Carcinoma (Endocrine Pancreas); Kaposi's Sarcoma; Kidney Cancer; Laryngeal Cancer; Laryngeal Cancer, Childhood; Leukemia, Acute Lymphoblastic, Adult; Leukemia, Acute Lymphoblastic, Childhood; Leukemia, Acute Myeloid, Adult; Leukemia, Acute Myeloid, Childhood; Leukemia, Chronic Lymphocytic; Leukemia, Chronic Myelogenous; Leukemia, Hairy Cell; Lip and Oral Cavity Cancer; Liver Cancer, Adult (Primary); Liver Cancer, Childhood (Primary); Lung Cancer, Non-Small Cell; Lung Cancer, Small Cell; Lymphoblastic Leukemia, Adult Acute; Lymphoblastic Leukemia, Childhood Acute; Lymphocytic Leukemia, Chronic; Lymphoma, AIDS-Related; Lymphoma, Central Nervous System (Primary); Lymphoma, Cutaneous T- Cell; Lymphoma, Hodgkin's, Adult; Lymphoma, Hodgkin's, Childhood; Lymphoma, Hodgkin's During Pregnancy; Lymphoma, Non-Hodgkin's, Adult; Lymphoma, Non- Hodgkin's, Childhood; Lymphoma, Non-Hodgkin's During Pregnancy; Lymphoma, Primary Central Nervous System; Macroglobulinemia, Waldenstrom's; Male Breast Cancer; Malignant Mesothelioma, Adult; Malignant Mesothelioma, Childhood; Malignant Thymoma; Medulloblastoma, Childhood; Melanoma; Melanoma, Intraocular; Merkel Cell Carcinoma; Mesothelioma, Malignant; Metastatic Squamous Neck Cancer with Occult Primary; Multiple Endocrine Neoplasia Syndrome, Childhood; Multiple Myeloma/Plasma Cell Neoplasm; Mycosis Fungoides; Myelodysplasia Syndromes; Myelogenous Leukemia, Chronic; Myeloid Leukemia, Childhood Acute; Myeloma, Multiple; Myeloproliferative Disorders, Chronic; Nasal Cavity and Paranasal Sinus Cancer; Nasopharyngeal Cancer; Nasopharyngeal Cancer, Childhood; Neuroblastoma; Non-Hodgkin's Lymphoma, Adult; Non-Hodgkin's Lymphoma, Childhood; Non-Hodgkin's Lymphoma During Pregnancy; Non-Small Cell Lung Cancer; Oral Cancer, Childhood; Oral Cavity and Lip Cancer; Oropharyngeal Cancer; Osteosarcoma/Malignant Fibrous Histiocytoma of Bone; Ovarian Cancer, Childhood; Ovarian Epithelial Cancer; Ovarian Germ Cell Tumor; Ovarian Low Malignant Potential Tumor; Pancreatic Cancer; Pancreatic Cancer, Childhood; Pancreatic Cancer, Islet Cell; Paranasal Sinus and Nasal Cavity Cancer; Parathyroid Cancer; Penile Cancer; Pheochromocytoma; Pineal and Supratentorial Primitive Neuroectodermal Tumors, Childhood; Pituitary Tumor; Plasma Cell Neoplasm/Multiple Myeloma; Pleuropulmonary Blastoma; Pregnancy and Breast Cancer; Pregnancy and Hodgkin's Lymphoma; Pregnancy and Non-Hodgkin's Lymphoma; Primary Central Nervous System Lymphoma; Primary Liver Cancer, Adult; Primary Liver Cancer, Childhood; Prostate Cancer; Rectal Cancer; Renal Cell (Kidney) Cancer; Renal Cell Cancer, Childhood; Renal Pelvis and Ureter, Transitional Cell Cancer; Retinoblastoma; Rhabdomyosarcoma, Childhood; Salivary Gland Cancer; Salivary Gland Cancer, Childhood; Sarcoma, Ewing's Family of Tumors; Sarcoma, Kaposi's; Sarcoma (Osteosarcoma)/Malignant Fibrous Histiocytoma of Bone; Sarcoma, Rhabdomyosarcoma, Childhood; Sarcoma, Soft Tissue, Adult; Sarcoma, Soft Tissue, Childhood; Sezary Syndrome; Skin Cancer; Skin Cancer, Childhood; Skin Cancer (Melanoma); Skin Carcinoma, Merkel Cell; Small Cell Lung Cancer; Small Intestine Cancer; Soft Tissue Sarcoma, Adult; Soft Tissue Sarcoma, Childhood; Squamous Neck Cancer with Occult Primary, Metastatic; Stomach (Gastric) Cancer; Stomach (Gastric) Cancer, Childhood; Supratentorial Primitive Neuroectodermal Tumors, Childhood; T-Cell Lymphoma, Cutaneous; Testicular Cancer; Thymoma, Childhood; Thymoma, Malignant; Thyroid Cancer; Thyroid Cancer, Childhood; Transitional Cell Cancer of the Renal Pelvis and Ureter; Trophoblastic Tumor, Gestational; Unknown Primary Site, Cancer of, Childhood; Unusual Cancers of Childhood; Ureter and Renal Pelvis, Transitional Cell Cancer; Urethral Cancer; Uterine Sarcoma; Vaginal Cancer; Visual Pathway and Hypothalamic Glioma, Childhood; Vulvar Cancer; Waldenstrom's Macroglobulinemia; and Wilms' Tumor. The methods of the present invention may be useful to treat such types of cancer.

In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the second active agent is an anti-cancer agent. In certain embodiments, the second active agent is a tyrosine kinase inhibitor or a Raf kinase inhibitor. In certain embodiments, the second active agent is imatinib, gefitinib, erlotinib, sunitinib, sorafenib, or quizartinib (AC220).

In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the cancer is breast cancer or ovarian cancer.

Another aspect of the invention relates to a method of treating or preventing acute pancreatitis, comprising the step of administering to a subject in need thereof a therapeutically effective amount of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby treating or preventing pancreatitis.

Pancreatitis is an inflammation of the pancreas mediated by the release of digestive enzymes that eventually lead to the destruction of the organ itself. Pancreatitis can be a severe, life-threatening illness with many complications. In severe cases, bleeding, tissue damage to the heart, lungs and kidneys, and infection may occur. About 80,000 cases of acute pancreatitis occur annually in the United States; about 20 percent of them are severe. There is no known treatment for pancreatitis. The current approaches for managing pancreatitis involve waiting for it to resolve on its own and the treatment of heart, lungs and kidney complications if that occur. Autophagy has been shown to play an important role in mediating cellular damage induced by acute pancreatitis. Autodigestion of the pancreas by its own prematurely activated digestive proteases is believed to be important for the onset of acute pancreatitis. Although lysosomal hydrolases are known to play a key role in pancreatic trypsinogen activation, it remains unclear where and how trypsinogen meets these lysosomal enzymes. Autophagy has been proposed to play a key role in the release of pancreatitic digestive enzymes in animal models of pancreatitis. In Atg5-/- mice, which are defective for a key autophagy gene Atg5, the severity of acute pancreatitis induced by cerulein is greatly reduced with a significantly decreased level of trypsinogen activation. Thus, activation of autophagy may exert a detrimental effect in pancreatic acinar cells by mediating the activation of trypsinogen to trypsin. Inhibition of autophagy may provide a unique opportunity for blocking trypsinogen activation in acute pancreatitis. Development of an autophagy inhibitor may provide a first-in-class inhibitor for acute pancreatitis.

Another aspect of the invention relates to a method of treating or preventing a disease caused by an intracellular pathogen, comprising the step of administering to a subject in need thereof a therapeutically effective amount of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby treating or preventing the disease caused by the intracellular pathogen.

Studies have established a role for autophagy in cellular defense against intracellular pathogens including bacteria, such as Mycobacterium tuberculosis, Streptococcus pyogenes, Shigella spp. and Salmonella typhimurium, as well as viruses and protozoa which use autophagosomes to proliferate. The execution of autophagy is regulated by upstream signal transduction systems that are influenced by largely physiological factors such as nutrient status, growth factors/cytokines, and hypoxia. The pharmacological induction of autophagy is a therapeutic strategy in which this effector of innate immunity would be triggered or amplified to defend against intracellular pathogens.

Another aspect of the invention relates to a method of treating or preventing a lysosomal storage disorder, comprising the step of administering to a subject in need thereof a therapeutically effective amount of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby treating or preventing the lysosomal storage disorder. Lysosomal storage diseases/disorders are a type of disease involving partial or complete deficiency of a lysosomal hydrolase. This deficiency results in incomplete lysosomal digestion of substrates specific to the hydrolase. Over time, the accumulation of undigested substrate can lead to various abnormalities, including progressive and severe neuro- and muscular-degeneration.

In certain embodiments, the lysosomal storage disorder is selected from the group consisting of GM2 Gangliosidosis, Alpha-mannosidosis, Aspartylglucosaminuria, Cholesteryl ester storage disease, Chronic Hexosaminidase A Deficiency, Cystinosis, Danon disease, Fabry disease, Farber disease, Fucosidosis, Galactosialidosis, Gaucher Disease, GMl gangliosidosis, I-Cell disease/Muco lipidosis II, Infantile Free Sialic Acid Storage Disease/ISSD, Juvenile Hexosaminidase A Deficiency, Krabbe disease, Metachromatic Leukodystrophy, Mucopolysaccharidoses disorders (Pseudo-Hurler polydystrophy/Mucolipidosis IIIA, MPSI Hurler Syndrome, MPSI Scheie Syndrome, MPS I Hurler-Scheie Syndrome, MPS II Hunter syndrome, Sanfilippo syndrome, Morquio Type A/MPS IV A, Morquio Type B/MPS IVB, MPS IX Hyaluronidase Deficiency, MPS VI Maroteaux-Lamy, MPS VII Sly Syndrome, Mucolipidosis I/Sialidosis, Mucolipidosis IIIC, Mucolipidosis type IV), Multiple sulfatase deficiency, Niemann-Pick Disease, Neuronal Ceroid Lipofuscinoses (CLN6 disease, Batten-Spielmeyer-Vogt/Juvenile NCL/CLN3 disease, Finnish Variant Late Infantile CLN5, Jansky-Bielschowsky disease/Late infantile CLN2/TPP1 Disease, Kufs/Adult-onset NCL/CLN4 disease, Northern Epilepsy/variant late infantile CLN8, Santavuori-Haltia/Infantile CLN1/PPT disease, Beta-mannosidosis), Pompe disease/Glycogen storage disease type II, Pycnodysostosis, Sandhoff disease, Schindler disease, Salla disease/Sialic Acid Storage Disease, Tay-Sachs/GM2 gangliosidosis, and Wolman disease.

Another aspect of the invention relates to a method of inhibiting autophagy in a cell in need thereof, comprising the step of contacting the cell with a therapeutically effective amount of any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby inhibiting autophagy in the cell.

Another aspect of the invention relates to a method of inactivating a deubiquitinating protease complex, comprising the step of contacting the deubiquitinating protease complex with any one of the aforementioned compounds, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, wherein the deubiquitinating protease complex comprises USP3 and USP10. Such methods can be used to ameliorate any condition that is caused by or potentiated by the activity of the deubiquitinating protease complex.

Combination Therapy

In one aspect of the invention, a compound of the invention, or a pharmaceutically acceptable salt thereof, can be used alone or in combination with another therapeutic agent to treat diseases such cancer and pancreatitis. It should be understood that the compounds of the invention can be used alone or in combination with an additional agent, e.g., a therapeutic agent, said additional agent being selected by the skilled artisan for its intended purpose. For example, the additional agent can be a therapeutic agent that is art-recognized as being useful to treat the disease or condition being treated by the compound of the present invention. The additional agent also can be an agent that imparts a beneficial attribute to the therapeutic composition e.g., an agent that affects the viscosity of the composition.

The combination therapy contemplated by the invention includes, for example, administration of a compound of the invention, or a pharmaceutically acceptable salt thereof, and additional agent(s) in a single pharmaceutical formulation as well as administration of a compound of the invention, or a pharmaceutically acceptable salt thereof, and additional agent(s) in separate pharmaceutical formulations. In other words, coadministration shall mean the administration of at least two agents to a subject so as to provide the beneficial effects of the combination of both agents. For example, the agents may be administered simultaneously or sequentially over a period of time.

It should further be understood that the combinations included within the invention are those combinations useful for their intended purpose. The agents set forth below are illustrative for purposes and not intended to be limited. The combinations, which are part of this invention, can be the compounds of the present invention and at least one additional agent selected from the lists below. The combination can also include more than one additional agent, e.g., two or three additional agents if the combination is such that the formed composition can perform its intended function.

While not wishing to be bound by any particular theory, it is possible that autophagy inhibitors used as a single agent are not going to be particularly useful therapeutically in cancer therapy. Cancer cells under the influence of chemotherapeutic agents will undergo autophagy, which helps cancer cells survive the damage caused by the chemotherapeutic agents. Autophagy inhibitors, such as those described herein, will stop the autophagy process and increase the effectiveness of chemotherapeutic agents, thereby displaying a synergistic effect.

In certain embodiments, the co-administration of two or more therapeutic agents achieves a synergistic effect, i.e., a therapeutic effect that is greater than the sum of the therapeutic effects of the individual components of the combination. The term "synergistic" refers to a combination which is more effective than the additive effects of any two or more single agents. A synergistic effect permits the effective treatment of a disease using lower amounts (doses) of individual therapy. The lower doses result in lower toxicity without reduced efficacy. In addition, a synergistic effect can result in improved efficacy. Finally, synergy may result in an improved avoidance or reduction of disease as compared to any single therapy. Combination therapy can allow for the product of lower doses of the first therapeutic or the second therapeutic agent (referred to as "apparent one-way synergy" herein), or lower doses of both therapeutic agents (referred to as "two-way synergy" herein) than would normally be required when either drug is used alone.

For example, one aspect of the invention relates to the use of small molecule autophagy inhibitors in combination with anti-angiogenesis inhibitors for the treatment of cancers. It is known that anti-angiogenesis inhibitors have the promise to inhibit tumor growth by suppressing the growth of blood vessels in tumors which are required for supporting tumor survival and growth. For example, the angiostatic agent endostatin and related chemicals can suppress the building of blood vessels and reduce tumor growth. Several hundred clinical trials of anti-angiogenesis drugs are now under way. In tests with patients, anti-angiogenesis therapies are able to suppress tumor growth with relatively few side effects. However, anti-angiogenesis therapy alone may not be insufficient to prolong patient survival; combination with a conventional chemotherapy may therfore be beneficial. Specifically, autophagy inhibitors may provide a new option to work alone or in combination with anti-angiogenesis therapy.

Endostatin has been shown to induce autophagy in endothelial cells by modulating Beclin 1 and beta-catenin levels (Nguyen, T.M., et al, Endostatin induces autophagy in endothelial cells by modulating Beclin 1 and beta-catenin levels. J Cell Mol Med, 2009). As disclosed herein, it has been found that inhibition of autophagy selectively kills a subset of cancer cells under starvation condition. Therefore, it is proposed that anti-angiogenesis therapy may induce additional metabolic stress to sensitize cancer cells to autophagy inhibitors, which are not normally cytotoxic. Thus, a combination of anti-angiogenesis therapy and anti-autophagy therapy may provide a new option for treatment of cancers without cytotoxicity to normal cells (Ramakrishnan, S., et al., Autophagy and angiogenesis inhibition. Autophagy, 2007. 3(5): p. 512-5).

Non-limiting examples of anti-angiogenesis agents with which a compound of the invention of the invention can be combined include, for example, the following: bevacizumab (Avastin®), carboxyamidotriazole, TNP-470, CMlOl, IFN-a, IL-12, platelet factor-4, suramin, SU5416, thrombospondin, VEGFR antagonists, angiostatic steroids with heparin, Cartilage-Derived Angiogenesis Inhibitory Factor, matrix metalloproteinase inhibitors, angiostatin, endostatin, 2-methoxyestradiol, tecogalan, thrombospondin, prolactin, νβ3 inhibitors and linomide.

In addition, as described in US Patent Application Publication No. 2008/0269259 to Thompson et al. (hereby incorporated by reference in its entirety), autophagy inhibitors can be used to treat a subject who has been identified as having a glycolysis dependent cancer by combining one or more autophagy inhibitors with one or more anti-cancer compounds which converts glycolysis dependent cancer to cells incapable of glycolysis. Examples of anti-cancer compounds which convert glycolysis dependent cancer to cells incapable of glycolysis: Alkylating Agents; Nitrosoureas; Antitumor Antibiotics; Corticosteroid Hormones; Anti-estrogens; Aromatase Inhibitors; Progestins; Anti-androgens; LHRH agonists; Kinase Inhibitors; and Antibody therapies; for example, busulfan, cisplatin, carboplatin, chlorambucil, cyclophosphamide, ifosfamide, dacarbazine (DTIC), mechlorethamine (nitrogen mustard), melphalan, carmustine (BCNU), lomustine (CCNU), dactinomycin, daunorubicin, doxorubicin (Adriamycin), idarubicin, mitoxantrone, prednisone, dexamethasone, tamoxifen, fulvestrant, anastrozole, letrozole, megestrol acetate, bicalutamide, flutamide. leuprolide, goserelin, gleevac, Iressa, Tarceva, Herceptin, Avastin, L-asparaginase and tretinoin.

Dosage

As used herein, a "therapeutically effective amount" or "therapeutically effective dose" is an amount of a compound of the invention or a combination of two or more such compounds, which inhibits, totally or partially, the progression of the condition or alleviates, at least partially, one or more symptoms of the condition. A therapeutically effective amount can also be an amount which is prophylactically effective. The amount which is therapeutically effective will depend upon the patient's size and gender, the condition to be treated, the severity of the condition and the result sought. For a given patient, a therapeutically effective amount can be determined by methods known to those of skill in the art.

For any compound used in a method of the present invention, the therapeutically effective dose can be estimated initially from cellular assays. For example, a dose can be formulated in cellular and animal models to achieve a circulating concentration range that includes the IC₅₀ as determined in cellular assays (i.e., the concentration of the test compound which achieves a half-maximal inhibition). In some cases it is appropriate to determine the IC₅₀ in the presence of 3 to 5% serum albumin since such a determination approximates the binding effects of plasma protein on the compound. Such information can be used to more accurately determine useful doses in humans.

A therapeutically effective dose refers to that amount of the compound that results in amelioration of symptoms in a patient. Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the maximum tolerated dose (MTD) and the ED₅₀ (effective dose for 50% maximal response). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio between MTD and ED₅₀. The data obtained from these cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED₅₀ with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition. (See e.g., Fingl et ah, 1975, in "The Pharmacological Basis of Therapeutics", Ch. 1 pi). In the treatment of crises, the administration of an acute bolus or an infusion approaching the MTD may be required to obtain a rapid response.

Dosage amount and interval may be adjusted individually to provide plasma levels of the active moiety which are sufficient to maintain the kinase modulating effects, or minimal effective concentration (MEC). The MEC will vary for each compound but can be estimated from in vitro data. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. However, HPLC assays or bioassays can be used to determine plasma concentrations. Dosage intervals can also be determined using the MEC value. Compounds should be administered using a regimen which maintains plasma levels above the MEC for 10-90% of the time, preferably between 30-90% and most preferably between 50-90% until the desired amelioration of symptoms is achieved. In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.

The amount of composition administered will, of course, be dependent on the subject being treated, on the subject's weight, the severity of the affliction, the manner of administration and the judgment of the prescribing physician.

Kits

The compounds and compositions of the invention may, if desired, be presented in a kit (e.g., a pack or dispenser device). The pack may for example comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for use of the compound in any method described herein. Compositions comprising a compound of the invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labelled for treatment of an indicated condition. Instructions for use may also be provided.

EXEMPLIFICATION

The invention now being generally described, it will be more readily understood by reference to the following examples, which are included merely for purposes of illustration of certain aspects and embodiments of the present invention, and are not intended to limit the invention.

Example 1: General Preparation of Compounds

One general approach to the synthesis of compounds of formula I, II, and III (Figure 2) is depicted in Figure 3.

[1] Step one is the formation of a quinazoline-4-ketone (or 8-aza- quinazoline-4-ketone)

In one approach, anthranilic acid is mixed with formamide in a molar 1 :25-30 and heated at about 200 °C for 20 hours. After the reaction is complete, the mixture is cooled, filtrated, washed with water and dried. The resulting crude product is used in the next steps without further processing.

[2] Step two is the formation of a 4-chloroquinazoline (or 8-aza- 4-chloroquinazoline)

In one approach, the crude product from step one is mixed with phosphorus oxycholoride in a molar 1 : 10 and heated at about 120 °C for 16 hours. After the reaction is complete, the mixture is cooled and excess phosphorus oxycholoride is removed by rotary evaporation. An organic solvent, such as dichloromethane, is added to dissolve the solid, followed by pH adjustment of resulting solution to about 7-8 by addition ammonia. The resulting mixture is extracted with dichloromethane, dried and purified by column chromatography.

In another approach, the crude product from step one is mixed with thionyl dichloride in a molar 1 :25, with catalytic amount of anhydrous DMF (e.g. 0.5-1 mL), then heated at about 100 °C for 16 hours. After the reaction is complete, the mixture is cooled and excess thionyl dichloride is removed by rotary evaporation. An organic solvent, such as dichloromethane, is added to dissolve the solid, and then petroleum ether (5 times the dichloromethane volume) is added to solution. The mixture is filtrated, washed with ether and petroleum ether, dried and purified by column chromatography.

[3] Step three is the formation of an N-substituted-4-amino-chloroquinazoline (or 8-aza- N- substituted-4-amino-chloroquinazoline)

Under nitrogen, the product of step 2, amine (as defined herein), and triethylamine are combined in a molar ratio of 1 : 1.2:2, in an organic solvent, such as isopropanol, and heated to about 50-80 °C for about 10-18 hours. After the reaction is complete, the mixture is cooled and organic solvent is removed by rotary evaporation. The resulting crude product is purified by Prep-HPLC.

For addition illustration, the synthesis of compounds is described in more detail below. As note above, additional compounds can be prepared by varying the amine which is coupled with optionally substituted 4-chloroquinazoline (such as Figure 4).

Example 2: Preparation of A02-172 (Figure 4)

The compound 172-1(1.55g, 10 mmol) is mixed with formamide (9.0 g, 0.2 mol) and heated at 200 °C for 20 hours. After the reaction is complete, the mixture is cooled, filtrated, washed with water and dried to afford crude product 172-2(0.98 g, yield: 60%). The resulting crude product is used in the next steps without further processing.

The crude product 172-2(0.82g, 5 mmol) is mixed with thionyl dichloride (12.0 g, 0.1 mol) and catalytic amount of anhydrous DMF (0.5 mL), then heated at about 100 °C for 16 hours. After the reaction is complete, the mixture is cooled and excess thionyl dichloride is removed by rotary evaporation. Dichloromethane (10 mL) is added to dissolve the solid, and then petroleum ether (50mL) is added to solution. The mixture is filtrated, washed with ether and petroleum ether, dried and purified by silica gel chromatography (PE:AE=1 : 1) to afford compound 172-3 as a white solid (0.75 g, yield: 82%).

To a solution of compound 172-3 (120 mg, 0.66 mmol) and Indan-2-amine (87 mg, 0.66 mmol) in isopropyl alcohol (5.0 mL) was added triethylamine (200 mg, 2.0 mmol). The resulting solution was heated to 65 °C under nitrogen for 8 hours. LC-MS analysis showed completed consumption of 172-3. The mixture is cooled and isopropyl alcohol is removed by rotary evaporation. The residue is purified by Prep-HPLC to give A02-172 as a white solid (103 mg, yield: 67%, confirmed by 1H NMR, and LC-MS). LC-MS 280 (M+H)⁺, Purity 100%(UV214); ¹HNMR (400 MHz, DMSO- 6) δ 8.517 (s, 1H), 8.22-8.28 (m, 2H), 7.75-7.78 (m, 1H), 7.67-7.70 (m, 1H), 7.25-7.28(m, 2H), 7.16-7.18 (m, 2H), 4.99 (d, J= 6.4Hz, 1H), 3.35-3.41(m, 2H), 3.01-3.07 (s, 2H).

Example 3: Preparation of A02-171 (Figure 5)

To a cold solution of K 0₃ (1.18 g, 11.183 mmol) in cone. H₂S0₄ (10 mL) was added slowly 2, 3-dihydro-lH-inden-2-amine hydrochloride (2.1 g, 12.426 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 60 min. The reaction mixture was added cold water and adjusted pH to 8 with cold 4.0 M aqueous NaOH. The water phase was extracted with PE (50 mL), EA (50 mL) and DCM (50 mL). The organics were dried over sodium sulfate, filtered and evaporated to give a crude product 1 which would be used in next step without further purification (1.2 g, yield: 55%).

To a solution of 4-chloro-6-fluoroquinazoline (360 mg, 2.31 mmol) in DMF (8.0 mL) was added compound 1 (360 mg, 2.31 mmol) and triethylamine (612 mg, 6.0 mmol). The mixture was stirred at 100 °C in microwave for 60 min. The resulting mixture was washed with water (15 mL), extracted with ethyl acetate (3 x 20 mL) and washed with brine (20 mL). The combined organic layer was dried over anhydrous sodium sulfate. The organic was evaporated to give a residue. The residue was purified by silica gel to afford the title compound 2 (180 mg, yield: 26%). LC-MS 325 (M+H)⁺, 90% (UV 214 nm).

To a solution of 6-fluoro-N-(5-nitro-2,3-dihydro-lH-inden-2-yl)quinazolin-4-amine (compound 2, 180 mg, 0.556 mmol) in MeOH (lOmL) was added 10% Pd/C (30 mg). The mixture was stirred at room temperature overnight under H₂ atmosphere. The resulting mixture was filtered through C elite and the filtrate was evaporated to give a residue. The residue was purified by Prep-HPLC to afford the desired product A02-171 (58 mg, yield: 36%). LC-MS 295(M+H)⁺, purity: 100%(UV 214 nm); HNMR (400 MHz, DMSO-d6) δ 8.49 (s, 1H), 8.26-8.23 (m, 2H), 7.78-7.64 (m, 2H), 6.90 (d, J = 8.4 Hz, 2H), 6.47 (s, 1H), 6.39 (d, J= 7.6Hz, 1H), 4.96-4.84 (m, 3H), 3.29-3.16 (m, 2H), 2.92-2.82 (m, 2H).

Example 4: Preparation of A23-001 (Figure 6)

To a solution of 4-chloro-6-iodoquinazoline (1.307g, 4.5 mmol) in DMF (12 mL) was added compound 1 (800 mg, 4.5 mmol) and triethylamine (1.36g, 13.5 mmol). The mixture was stirred at 120 °C in microwave for 60 min. The resulting mixture was washed with water (15 mL), extracted with ethyl acetate (3 x 20 mL) and washed with brine (20 mL). The combined organic layer was dried over anhydrous sodium sulfate. The organic was evaporated to give a residue. The residue was purified by silica gel to afford A29-003 (610mg, yield: 31%). LC-MS 433 (M+H) ⁺, purity 95 % ( UV 214 nm).

In another approach, to a solution of 4-chloro-6-iodoquinazoline (1.23 mg, 4.2 mmoL) in isopropanol (30 mL was added compound 1 (631 mg, 3.5 mmol) and triethylamine (0.5 g, 5.0 mmol). The mixture was stirred at 70 for 8 h. The mixture is cooled down and isopropanol is removed by rotary evaporation. The residue was purified by silica gel to give desired product A29-003 (923 mg, yield: 50 %) as a yellow solid. LCMS (m/z): 433.0 (M+H)⁺. ¹FiNMR (400 MHz, DMSO- 6): δ 8.75 (s, 1H), 7.56 (s, 1H), 8.46 (d, J = 6.4 Hz, 1H), 8.15 (s, 1H), 8.09 (d, J = 6.4 Hz, 1H), 8.03 (d, J = 7.2 Hz, 1H), 7.55 (d, J = 8.4 Hz, 1H), 7.48 (d, J = 8.8 Hz, 1H), 5.08-5.06 (m, 2H), 3.53-3.47 (m, 2H), 3.19-3.14 (m, 2H).

A mixture of A29-003 (260 mg, 0.602 mmol), Zn(CN)₂ (139 mg, 1.204 mmol), Zn (10 mg), PdCl₂dppf (30 mg) and DMF (3 mL) was heated at 145°C by microwave for 150 min. The mixture was cooled down to room temperature and washed with NH₄OH, extracted with ethyl acetate. The organic layer was dried over sodium sulfate, filtered and evaporated to give a residue. The residue was purified by silica gel to give A29-005 (200 mg, yield: 100%). LC-MS 332 (M+H) ⁺, purity 92 % ( UV 214 nm).

In another approach, a mixture of compound A29-003 (2.00 g, 4.63 mmol) and CuCN (2.06 g, 23.15 mmol) in DMSO (25 mL) was heated to 148°C for 8 hours. After the mixture was added ammonia (5.0 mL) and ethyl acetate (100 mL). The organic layer was washed successively with water (2 x 20 mL), brine (2 x 20 mL) and dried over sodium sulfate, filtered and evaporated to give a yellow solid. The crude product was purified by silica gel to give A29-005 (900 mg, yield: 59 %) as a yellow solid. LCMS 332 (M+H) ⁺. ¹HNMR (400 MHz, DMSO- 6): δ 8.93 (s, 1H), 8.67 (d, J = 6.0 Hz, 1H), 8.65 (s, 1H), 8.17 (s, 1H), 8.09 (t, J = 8.4 Hz, 2H), 7.80 (d, J = 8.8 Hz, 1H), 7.56 (d, J = 8.4 Hz, 1H), 5.10- 5.08 (m, 1H), 3.55-3.49 (m, 2H), 3.20-3.15 (m, 2H).

A mixture of A29-005 (200 mg, 0.60 mmol), Zn (249 mg, 3.8 mmol) and NH₄C1 (340 mg, 6.4 mmol) in MeOH (10 mL) was stirred at 60 °C for 18 h. The mixture was concentrated and extracted with ethyl acetate, washed with water and brine, dried by sodium sulfate. The residue was purified by Prep-HPLC to give the title compound A23- 001 (61 mg, yield: 31%). LC-MS 302(M+H)⁺, purity 93%(UV 214 nm); ¹HNMR (400 MHz, CD₃OD) δ 8.66 (d, J = 1.6Hz, 1H), 8.47 (s, 1H), 7.89-7.86 (m, 1H), 7.68 (d, J = 8.8 Hz, 1H), 6.90 (d, J = 8.0 Hz, 2H), 6.57 (s, 1H), 6.51-6.48 (m, 1H), 5.02-4.98 (m, 1H), 3.26- 3.17 (m, 2H), 2.91-2.81(m, 2H).

Example 5: Inhibition of Autophagy

1) Tetrahydronaphthalene series:

These groups of compounds are similar to linear two-carbon linker series of compounds in terms of linker length, but in this case the linker is part of saturated ring of tetrahydronaphthtalene. They should have a smaller possible number of conformations. If their more stable conformations are the active ones, binding protein target binding sites, they should bind more favorably entropically than the compounds with linear linkers. In addition, these compounds could be potentially more selective towards desired target, and consequently cause less off-target toxicity in in vivo assay. Over thirty of these compounds have been prepared. A few compounds are summarized in Figure 8.

2) Indene series:

Similar to tetrahydronaphthalene-containing series, the indene-containing series has a constrained linker between core structure of quinazoline and right hand benzene ring. Furthemore, this class of compounds is more potent and stable compared to the tetrahydrohapphthalene-containing series. For example, compounds A02-171, A23-001 and A23-024 in Figure 9 have EC50s below or equal to 0.05 uM. And these compounds also have improved mouse microsomal stability. Compound A29-010 is stable in microsomal and its T 1/2 is 163 minutes.

3) Tetrahydrobenzothiazole series

A series of tetrahydrobenzothiazole-containing compounds has shown good potency in a cellular autophagy assay. 3-Aminothiazole analogue A02-189 (Figure 10) has EC50 = 0.59 uM and the amide version analogues of this aminothiazole (A31-001, A31-002, A31- 003, A31-004, A31-005 and A31-006) all have good autophagy potency and more importantly, these amide derivatives of aminothiazole show much improved mouse mocrosomal stability. For example, A31-001, 2-hydroxyacetamide derivative of 3- aminothiazole A02-189 is the most stable one in this series (Tl/2 = 125.6 minutes), and A31-00, an isobutyramide derivative of A02-189 has a Tl/2 = 107.9 minutes.

Example 6

1) . Combination effect of lead compounds with AC220 in p53 mutant breast and ovarian cancer cell lines

Since p53 mutant breast and ovarian cancer cell lines are sensitive to autophagy inhibition, four p53 mutant breast cancer cell lines (BT-474, HCC70, MDA-MB-231 and SUM 159) and four p53 mutant ovarian cancer cell lines (OVCAR-3, Caov-3, SK-OV-3 and ES-2) were selected. AC220 was chosen for the combination assay. First, he combination assay of 4 potent compounds (A70, A22-022, A22-029 and A02-121) was performed with AC220 in these 8 cancer cell lines. The results showed that compounds and AC220 alone did not exhibit significant inhibitory effect on ES-2 and SUM 159 cell proliferation. AC220 sensitized ES-2 and Suml59 cells to the growth inhibition by the inhibitors (Figure 11). ES- 2 and SUM 159 were chosen as the model cell lines for the later experiments. Another 15 selected compounds were tested and combined with AC220 in ES-2 and SUM159 cells, among which A31-001, A31-003, A31-005 and A29-002 were chosen as the current lead compounds to perform acute tox and PK study (Figure 12 and Figure 13).

2) . Inhibitory effect of potent compounds on autophagy marker

Given that LC3B-II is a marker protein of autophagy, LC3B-II protein level were detected in ES-2 and SUM159 cells after treatment with AC220 and some selected compounds. AC220 increased LC3B-II protein level in ES-2 and SUM 159 cells at different time points, suggesting that AC220 induced autophagy in these two cells. A02-121, A23- 001 and A02-171 decreased AC220-induced LC3B-II, while A22-022 showed no effect on AC220-induced LC3B-II level.

3) . Compound profiling in breast and ovarian cell lines in glucose free condition

To detect if autophagy inhibitors can be used as monotherapy, A22-022, A02-171 and A23-001 were profiled in SUM159, SK-OV-3, SW620 and A2780 cells in normal and glucose free (GF) culture condition (Figure 14). Absolute IC50 of A02-171 and A23-001 in A2780 and SW620 cells shifted in glucose free culture condition. Example 7: Preparation compound A23-024 (Figure 15)

To a solution of A23-001 (150 mg, 0.50 mmol) in DMF (2.0 mL) was added 2- hydroxyacetic acid (46 mg, 0.6 mmol), HATU (285 mg, 0.75 mmol), triethylamine (0.1 ml, 0.72 mmol). The reaction mixture was stirred at 50 °C overnight. The resulting mixture was concentrated by rotary evaporation and the residue was purified by Prep-HPLC to give A23-024 as a white solid (20 mg, yield: 11%). LC-MS 360 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.56 (s, 1 H), 8.98 (s, 1 H), 8.67 (d, J = 6.0 Hz, 1 H), 8.64 (d, J = 11.2 Hz, 1 H), 8.07 (d, J = 7.2 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.66 (s, 1 H), 7.46 (d, J = 4.0 Hz, 1 H), 7.19 (d, J = 8.0 Hz, 1 H), 5.65 (t, J = 6.0 Hz, 1 H), 5.04-4.99 (m, 1 H), 3.97 (d, J = 6.0 Hz, 2 H), 3.39-3.35 (m, 2 H), 3.06-2.96 (m, 2 H).

Example 8: Preparation of compound A29-001 (Figure 16)

To a solution of A23-001 (150 mg, 0.50 mmol) in DMF (2.0 mL) was added 2- (dimethylamino) acetic acid (62 mg, 0.60 mmoL), HATU (285 mg, 0.75 mmol), triethylamine (0.10 mL, 0.72 mmol). The mixture was stirred at 50 °C overnight. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29- 001 as a white solid (35 mg, yield: 18%). LC-MS 387 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.78 (s, 1 H), 8.61 (s, 1 H), 8.01 (d, J = 6.8 Hz, 1 H), 7.81 (d, J = 8.8 Hz, 1 H), 7.56 (s, 1 H), 7.35 (d, J = 6.0 Hz, 1 H), 7.23 (d, J = 8.0 Hz, 1 H), 5.18-5.15 (m, 1 H), 3.47-3.34 (m, 2 H), 3.15 (s, 2 H), 3.12-3.08 (m, 2 H), 2.39 (s, 6 H). Example 9: Preparation of compound A29-002 (Figure 17)

To a solution of A23-001 (150 mg, 0.50 mmol) in DMF (2.0 mL) was added 3- hydroxy-3-methylbutanoic acid (71 mg, 0.60 mmoL), HATU (285 mg, 0.75 mmol), triethylamine (0.10 mL, 0.72 mmol). The reaction mixture was stirred at 80 °C overnight. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-002 as a white solid (25 mg, yield: 13%). LC-MS 402 (M+H) ⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.78 (s, 1 H), 8.61 (s, 1 H), 8.01(d, J = 6.8 Hz, 1 H), 7.82 (d, J = 4.8 Hz, 1 H), 7.54 (s, 1 H), 7.31 (d, J = 8.4 Hz, 1 H), 7.22 (d, J = 4.8 Hz, 1 H), 5.19-5.15 (m, 1 H), 3.49-3.41 (s, 2 H), 3.14-3.05 (m, 2 H), 2.54 (s, 2 H), 1.35 (s, 6 H). Example 10: Preparation of compound A29-007 (Figure 18)

To a solution of A23-001 (150 mg, 0.48 mmol) in dichloromethane (10 mL) was added isobutyryl chloride (58 mg, 0.55 mmoL), triethylamine (0.7 mL, 5.0 mmol). The reaction mixture was stirred at room temperature overnight. Dichloromethane was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-007 as a white solid (30 mg, yield: 16 %). LC-MS 372 (M+H)⁺, Purity 100% (UV 214 nm); 1H NMR(400 MHz, DMSO-d6) δ 9.74 (s, 1 H), 8.98 (s, 1 H), 8.66 (d, J = 6.4 Hz, 1 H), 8.63 (s, 1 H), 8.07 (d, J = 7.2 Hz, 1 H), 7.79 (d, J = 8.4 Hz, 1 H), 7.60 (s, 1 H), 7.36 (d, J = 6.8 Hz, 1 H), 7.17 (d, J = 8.4 Hz, 1 H), 5.04-4.99 (m, 1 H), 3.38-3.30 (m, 2 H), 3.05-2.96 (m, 2 H), 259-2.54 (m, 1 H), 1.09 (d, J = 7.2 Hz, 6 H).

Example 11: Preparation of compound A29-008 (Figure 19)

To a solution of A23-001 (150 mg, 0.48 mmol) in dichloromethane (10 mL) was added cyclopropanecarbonyl chloride (57 mg, 0.55 mmoL), triethylamine (0.7 mL, 5.0 mol). The reaction mixture was stirred at room temperature overnight. Dichloromethane was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-

008 as a white solid (72 mg, yield: 36%). LC-MS 340 (M+H)⁺, Purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 10.10 (s, 1 H), 8.98 (s, 1 H), 8.66 (d, J = 6.8 Hz, 1 H), 8.62 (s, 1 H), 8.06 (d, J = 6.8 Hz, 1 H), 7.79 (d, J = 8.4 Hz, 1 H), 7.57 (s, 1 H), 7.35 (d, J = 8.0 Hz, 1 H), 7.17 (d, J = 8.0 Hz, 1 H), 5.03-4.99 (m, 1 H), 3.38-3.30 (m, 2 H), 3.04-2.95 (m, 2 H), 1.77-1.75 (m, 1 H), 0.77 (d, J = 2.4 Hz, 4 H).

Example 12: Preparation of compound A29-009 (Figure 20)

To a solution of A23-001 (130 mg, 0.43 mmol) in dichloromethane (10 mL) was added 3, 3-dimethylbutanoyl chloride (63 mg, 0.47 mmoL), triethylamine (0.7 mL, 5.0 mmol). The reaction mixture was stirred at room temperature overnight. Dichloromethane was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-

009 as a white solid (27 mg, yield: 16%). LC-MS 400 (M+H)⁺, Purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.71 (s, 1 H), 8.97 (s, 1 H), 8.66 (d, J = 6.0 Hz, 1 H), 8.62 (s, 1 H), 8.06 (d, J = 7.2 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.60 (s, 1 H), 7.32 (d, J = 8.4 Hz, 1 H), 7.17 (d, J = 8.0 Hz, 1 H), 5.03-4.99 (m, 1 H), 3.38-3.30 (m, 2 H), 3.05-2.95 (m, 2 H), 2.17 (s, 2 H), 0.98 (s, 2 H).

Example 13: Preparation of A29-011 (Figure 21)

To a solution of A23-001 (130 mg, 0.43 mmol) in dichloromethane (10 mL) was added methanesulfonyl chloride (54 mg, 0.47 mmoL), triethylamine (0.7 mL, 5.0 mmol). The reaction mixture was stirred at room temperature overnight. Dichloromethane was removed by rotary evaporation. The residue was dissolved in THF (5.0 mL) and treated with sodium hydroxide solution (3 M, 4.0 mL) at 25 °C for 2 h. THF was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-011 as a white solid (40 mg, yield: 25%). LC-MS 380 (M+H)⁺, Purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.58 (s, 1 H), 8.97 (s, 1 H), 8.66 (d, J = 6.4 Hz, 1H), 8.62 (s, 1 H), 8.07 (d, J = 7.2 Hz, 1 H), 7.79 (d, J = 8.4 Hz, 1 H), 7.23 (d, J = 8.0 Hz, 1 H), 7.13 (s, 1 H), 7.04 (d, J = 7.6 Hz, 1 H), 5.04-4.99 (m, 1 H), 3.40-3.36 (m, 2 H), 3.06-2.95 (m, 2 H), 2.50 (s, 3 H).

Example 14: Preparation of A29-012 (Figure 22)

To a solution of A23-001 (130 mg, 0.43 mmol) in dichloromethane (10 mL) was added 2-methoxyacetyl chloride (51 mg, 0.47 mmoL), triethylamine (0.7 mL, 5.0 mmol). The reaction mixture was stirred at room temperature overnight. Dichloromethane was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-012 as a white solid (25 mg, yield: 16%). LC-MS 374 (M+H)⁺, Purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.67 (s, 1 H), 8.98 (s, 1 H), 8.68 (d, J = 6.4 Hz, 1 H), 8.63 (s, 1 H), 8.07 (d, J = 7.2 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.63 (s, 1 H), 7.43 (d, J = 6.4 Hz, 1 H), 7.19 (d, J = 8.8 Hz, 1 H), 5.04-4.99 (m, 1 H), 4.00 (s, 2 H), 3.41-3.29 (m, 5 H), 3.06-2.96 (m, 2 H).

Example 15: Preparation of A29-013 (Figure 23)

To a solution of A23-001 (150 mg, 0.48 mmol) in dichloromethane (15 mL) was added 2-methoxyethanesulfonyl chloride (158 mg, 1.0 mmol), triethylamine (0.7 mL, 5.0 mmol). The mixture was stirred at room temperature for overnight. Dichloromethane was removed by rotary evaporation. The residue was dissolved in THF (5.0 mL) and treated with sodium hydroxide solution (3 M, 4.0 mL) at 25 °C for 2 h. THF was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-013 as a white solid (25 mg, yield: 16%). LC-MS 424 (M+H)⁺, Purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.98 (s, 1 H), 8.67 (d, J = 6.0 Hz, 1 H), 8.62 (s, 1 H), 8.07 (d, J = 7.2 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.21 (d, J = 8.0 Hz, 1 H), 7.12 (s, 1 H), 7.02 (d, J = 6.4 Hz, 1 H), 5.02-4.99 (m, 1 H), ), 3.65 (t, j = 6.4 Hz, 2 H), 3.31-3.20 (m, 4 H), 3.16 (s, 3 H), 3.05-2.96 (m, 2 H).

Example 16: Preparation of compound A29-014 (Figure 24)

To a solution of A23-001 (150 mg, 0.50 mmol) in DMF (2.0 mL) was added 2- morpholinoacetic acid (145 mg, 1.0 mmoL), HATU (380 mg, 1.0 mmol), triethylamine (0.10 mL, 0.72 mmol). The reaction mixture was stirred at 50 °C for 20 h. The resulting mixture was concentrated by rotary evaporator and the residue was purified by Prep-HPLC to give A29-014 as a white solid (15 mg, yield: 7%). LC-MS 429 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.66 (s, 1 H), 8.97 (s, 1 H), 8.67 (d, J = 6.0 Hz, 1 H), 8.62 (s, 1 H), 8.07 (d, J = 6.8 Hz, 1 H), 7.79 (d, J = 8.4 Hz, 1 H), 7.60 (s, 1 H), 7.40 (d, J = 6.8 Hz, 1 H), 7.19 (d, J = 7.6 Hz, 1 H), 5.03-4.99 (m, 1 H), ), 3.64 (t, J = 4.4 Hz, 4H), 3.39-3.30 (m, 6 H), 3.1 1 (s, 2 H), 3.06-2.96 (m, 2 H).

Example 17 Preparation of A29-006 (Figure 25)

A mixture of compound A29-003 (2.00 g, 4.63 mmol), H₂0 (0.1 mL, 5.5 mmol) and CuCN (2.06 g, 23.15 mmol) in DMSO (25 mL) was heated to 148 °C for 8 h. After the reaction mixture was diluted with ammonium hydroxide (5.0 mL), the mixture was extracted with ethyl acetate (100 mL). The organic layer was washed successively with water (2 x 20 mL) and brine (2 x 20 mL), dried over sodium sulfate, filtered and concentrated to give a yellow solid. The crude product was purified by silica gel column chromatography (using petroleum ether : ethyl acetate = 1 : 1 - 3 :7) give A29-006 (1.0 g, yield: 62%) as a yellow solid. LCMS 350 (M+H)⁺. 1H NMR (400 MHz, DMSO- 6): δ 8.85 (s, 1 H), 8.62 (d, J = 6.4 Hz, 1 H), 8.57 (s, 1 H), 8.18 (d, J = 6.8 Hz, 2 H), 8.09 (d, J = 6.4 Hz, 1 H), ), 8.01 (s, 1 H), 7.71 (d, J = 8.8 Hz, 1 H), 7.55 (d, J = 6.3 Hz, 1 H), 5.15-5.10 (m, 1 H), 3.54-3.48 (m, 2 H), 3.23-3.18 (m, 2 H).

Example 18: Preparation of A29-010 (Figure 26)

A mixture of A29-006 (130 mg, 0.37 mmol), Zn (118 mg, 1.85 mmol) and NH₄C1 (158 mg, 3.0 mmol) in MeOH (5.0 mL) was stirred at 70 °C for 8 h. After cooled down, the reaction mixture was concentrated with rotary evaporator. The crude product was extracted with ethyl acetate. The organic phase was washed with water (10 mL) and brine (10 mL), dried over sodium sulfate, filtered and concentrated. The residue was purified by Prep- HPLC to give the title compound A29-010 (25 mg, yield: 21%). LC-MS 320 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6): δ 8.87 (s, 1 H), 8.53 (s, 1 H), 8.48 (d, J = 6.8 Hz, 1 H), 7.95 (s, 1 H), 7.69 (d, J = 8.4 Hz, 1 H), 7.55 (s, 1 H), 6.89 (d, J = 8.0 Hz, 1 H), 6.47 (s, 1 H), 6.39 (d, J = 7.6 Hz, 1 H), 5.03-4.99 (m, 1 H), 4.84 (s, 1 H), 3.31-3.17 (m, 2 H), 2.96-2.86 (m, 2 H).

Example 19: Preparation of A26-010 (Figure 27)

To a solution of 4, 6-dichloroquinazoline (200 mg, 1.01 mmol) in isopropanol (30 mL) was added compound 1 (150 mg, 0.84 mmol) and triethylamine (0.5 g, 5.0 mmol). The reaction mixture was stirred at 70 °C for 8 h. After the reaction mixture was cooled down, isopropanol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give the title compound A29-004 (53 mg, yield: 19%). LC-MS 341 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6): δ 8.55 (s, 1 H), 8.49 (d, J = 2.4 Hz, 1 H), 8.45 (d, J = 6.0 Hz, 1 H), 8.16 (s, 1 H), 8.09 (d, J = 6.0 Hz, 1 H), 7.79 (d, J = 6.8 Hz, 1 H), 7.71(d, J = 9.2 Hz, 1 H), 7.55 (d, J = 8.4 Hz, 1 H), 5.08-5.05 (m, 1 H), 3.53-3.47 (m, 2 H), 3.19-3.14 (m, 2 H).

A mixture of A29-004 (160 mg, 0.47 mmol), Zn (151 mg, 2.35 mmol) and NH₄C1 (238 mg, 3.8 mmol) in MeOH (15 mL) was stirred at reflux for 2 h. After cooled down, the reaction mixture was concentrated. The crude product was extracted with dichloromethane (2 x 50 mL). The organic phase was washed with water (10 mL) and brine (10 mL), dried over sodium sulfate, filtered and concentrated. The residue was purified by Prep-HPLC to give the title compound A26-010 (25 mg, yield: 18%). LC-MS 311 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6): δ 8.48 (s, 1 H), 8.34 (s, 1 H), 7.76-7.67 (m, 2 H), 6.99 (d, J = 6.0 Hz, 1 H), 6.68 (s, 1 H), 6.61 (d, J = 4.5 Hz, 1 H), 5.08-5.04 (m, 1 H), 3.36-3.28 (m, 2 H), 3.01-2.92 (m, 2 H).

Example 20: Preparation of A02-188 (Figure 28)

To a solution of 5-fluoro-2, 3-dihydroinden-l-one (600 mg, 4.0 mmol) in ethnol (20 mL) was added sodium acetate (820 mg, 10.0 mmol) and a solution of hydroxylamine hydrochloride (690 mg, 10 mmol) in water (10 mL). The reaction mixture was re fluxed at 100 °C for 4.0 h under nitrogen atmosphere. After the reaction mixture was cooled down, excess ethanol was removed by rotary evaporation. The crude product was extracted with ethyl acetate (2 x 50 mL). The organic layer was washed successively with water (2 x 20 mL), brine (2 x 20 mL), dried over sodium sulfate, filtered and concentrated to give a white solid. The crude product was purified by silica gel column chromatography (using petroleum ether : ethyl acetate = 9: 1 - 7:3) to give compound 2 (650 mg, yield: 98.5%>). LC-MS 166 (M+H)⁺, purity 100% (UV 214 nm).

To a solution of compound 2 (650 mg, 3.93 mmol) in MeOH (30 mL) was added Pd/C (200 mg, 10%) and acetate acid (3.0 mL). The reaction mixture was stirred at room temperature overnight under hydrogen atmosphere. The resulting mixture was filtered and the filtrate was evaporated to give the crude product which was purified by silica gel column chromatography (using petroleum ether : ethyl acetate = 2: 1 - 1 : 1) to give compound 3 (600 mg, yield: 99%) as a sticky oil. LC-MS 152 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6): δ 7.40-7.33 (m, 1 H), 7.03-6.97 (m, 2 H), 6.36- 6.26 (s, 2 H), 4.27-4.23 (m, 1 H), 2.92-2.85 (m, 1 H), 2.76-2.68 (m, 1 H), 2.40-2.30 (m, 1 H), 1.84-1.65 (m, 1 H). To a solution of 4, 6-dichloroquinazoline (182 mg, 1.0 mmol) in isopropanol (10 mL) was added compound 3 (151 mg, 1.0 mmol) and triethylamine (0.7 mL, 5.0 mmol). The reaction mixture was stirred at 80 °C for 8 h. After the reaction mixture was cooled down, isopropanol was removed by rotary evaporation. The residue was purified by Prep- HPLC to give the title compound A02-188 (35 mg, yield: 12%). LC-MS 298 (M+H)+, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6): δ 8.52 (s, 1 H), 8.42-8.40 (m, 1 H), 8.23-8.19 (m, 1 H), 7.80-7.78 (m, 1 H), 7.72-7.66 (m, 1 H), 7.28-7.25 (m, 1 H), 7.16- 7.13 (m, 1 H), 7.00-6.96 (m, 1 H), 5.96-5.94 (m, 1 H), 3.05-3.02 (m, 1 H), 2.93-2.90 (m, 1 H), 2.60-2.49 (m, 1 H), 2.13-2.07 (m, 1 H).

Example 21: Preparation of A28-007 (Figure29)

To a solution of 4-chloro-6-iodoquinazoline (870 mg, 3.0 mmol) in isopropanol (10 mL) was added compound 3 (453 mg, 3.0 mmol) and triethylamine (0.7 mL, 5.0 mmol). The reaction mixture was stirred at 80 °C for 3 h. After thereaction mixture was cooled down, isopropanol was removed by rotary evaporation. The residue was purified by Prep- HPLC to give the title compound A28-007 (320 mg, yield: 26%). LC-MS 406 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, MeOH- 4): δ 8.68 (s, 1 H), 8.53 (s, 1 H), 8.09-8.06 (m, 1 H), 7.50 (d, J= 8.8, 1 H), 7.31-7.21 (m, 1 H), 7.03-7.01 (m, 1 H), 6.95-6.91 (m, 1 H), 6.02 (t, J = 7.2, 1 H), 3.12-3.10 (m, 1 H), 2.98-2.96 (m, 1 H), 2.72-2.67 (m, 1 H), 2.17-2.13 (m, 1 H).

Example 22: Preparation of A23-019 (Figure 30)

To a solution of A28-007 (120 mg, 0.30 mmol) in dry DMA (5.0 mL) was added CuCN (110 mg, 1.2 mmol). The reaction mixture was stirred at 120 °C for 20 h under nitrogen atmosphere. The resulting mixture was diluted with water (20 mL), extracted with EtOAc (3 x 30 mL) and washed with water (20 mL) and brine (20 mL). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by Prep-HPLC to give the title compound A29-019 (20 mg, yield: 22 %) as a white solid. LC-MS 305 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSC /6): δ 8.97 (s, 1 H), 8.81 (m, 1 H), 8.63 (s, 1 H), 8.10-8.07 (m, 1 H), 7.81 (d, J = 8.8, 1 H), 7.32-7.29 (m, 1 H), 7.17-7.14 (m, 1 H), 7.01-6.99 (m, 1 H), 5.95 (t, J = 7.2, 1 H), 3.06-3.05 (m, 1 H), 2.92-2.89 (m, 1 H), 2.61-2.58 (m, 1 H), 2.14-2.08 (m, 1 H).

Example 23: Preparation of A22-146 (Figure 31)

To a solution of A28-007 (121 mg, 0.3 mmol) in dry DMSO (3.0 mL) were added Cul (5.7 mg, 0.03 mmol) and sodium methanesulfmate (141 mg, 1.2 mmol). The reaction mixture was stirred at 120 °C for 2 h in microwave. The resulting mixture was diluted by water (20 mL), extracted with ethyl acetate (2 x 50 mL) and washed with water (20 mL) and brine (20 mL). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by Prep-HPLC to give the title compound A22-146 (25 mg, yield: 23%) as a white solid. LC-MS 358 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6): δ 9.10 (s, 1 H), 9.08 (s, 1 H), 8.64 (s, 1 H), 8.21-8.19 (m, 1 H), 7.90-7.87 (m, 1 H), 7.31-7.28 (m, 1 H), 7.17-7.14 (m, 1 H), 7.02- 6.97 (m, 1 H), 6.01 (t, J = 7.2, 1 H), 3.27 (s, 1 H), 3.07-3.04 (m, 1 H), 2.94-2.89 (m, 1 H), 2.61-2.49 (m, 1 H), 2.17-2.08 (m, 1 H).

Example 24: Preparation of A02-189 (Figure 32)

To a solution of 4-acetamidocyclohexanone (3.1 g, 20 mmol) in 30 mL of AcOH was added Br₂ (3.2 g, 20 mmol) at 60 °C with stirring. After continuous stirring for 1 h, thiourea (3.0 g, 40 mmol) was added. The reaction mixture was refluxed for 1 h. After cooled down, the reaction mixture was concentrated by rotary evaporator, diluted with 20 mL of water. Then pH value was adjusted to 10 with 50%> aqueous (aq.) NaOH solution. The precipitate was filtered off and washed with water (2 x 5 mL) and CH3OH (5 mL) to provide compound 4 (2.6 g, yield: 46%). LC-MS 212 (M+H)⁺, purity 92%(UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 7.08 (d, J = 8 Hz, 1 H), 5.62 (s, 2 H), 4.22 (m, 1 H), 2.84 and 2.41 (m, 2 H), 2.56 (m, 2 H), 1.68-2.08 (m, 2 H), 1.90 (s, 3 H).

A mixture of compound 4 (7.2 g, 25 mmol) and 47% aqueous HBr (72 mL) was refluxed for 20 h. After cooled down, the precipitate was formed and was filtered off. The precipitate was washed with cold water (5 mL) and acetone (5 mL) to provide 7.8 g of product 5. The resulting product was used for the next step reaction without further purification.

To a solution of compound 4-chloro-6-fluoroquinazoline (182 mg, 1.0 mmol) and compound 5 (333 mg, 1.0 mmol) in isopropyl alcohol (10 mL) was added triethylamine (1.0 mL, 7.2 mmol). The resulting solution was heated to 70 °C under nitrogen overnight. LC-MS analysis showed completed consumption of compound 5. After the reaction mixture was cooled down, excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A02-189 as a yellow solid (302 mg, yield: 95%). LC- MS 316 (M+H)⁺, purity 98% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.49 (s, 1 H), 8.24-8.28 (m, 1 H), 8.15 (d, J = 7.6 Hz, 1 H), 7.66-7.78 (m, 2 H), 6.77-6.81 (m, 2 H), 4.579 (s, 1 H), 2.93-2.98 (m, 1 H), 2.57-2.66 (m, 3 H), 2.06-2.09 (m, 1 H), 1.87-1.97 (m, 1 H).

Example 25: Preparation of A26-011 (Figure 33)

To a solution of compound 4,6-dichloroquinazoline (199 mg, 1.0 mmol) and 4,5,6,7-tetrahydrobenzolthiazole-2,6-diamine (333 mg, 1.0 mmol) in isopropyl alcohol (5.0 mL) was added triethylamine (1.0 mL, 7.2 mmol). The resulting solution was heated to 70 °C under nitrogen overnight. LC-MS analysis showed completed consumption of starting material. The mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A26-011 as a yellow solid (307 mg, yield: 92%). LC-MS 332 (M+H)⁺, 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.54 (d, J = 2.0 Hz, 1 H), 8.50 (s, 1 H), 8.26 (d, J = 7.6 Hz, 1 H), 7.69-7.80 (m, 2 H), 6.77 (s, 2 H), 4.58 (s, 1 H), 2.93-2.98 (m, 1 H), 2.57-2.65 (m, 3 H), 2.05-2.08 (m, 1 H), 1.88-1.97 (m, 1 H).

Example 26: Preparation of A28-008 (Figure 34)

To a solution of compound 4-chloro-6-iodoquinazoline (1.10 g, 3.8 mmol) and 4,5,6,7-tetrahydrobenzothiazole-2,6-diamine (1.26 g, 3.8 mmol) in isopropyl alcohol (20 mL) was added triethylamine (5.0 mL, 36 mmol). The resulting solution was heated to 70 °C under nitrogen overnight. LC-MS analysis showed completed consumption of starting material. The mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A28-008 as a yellow solid (1.5 g, yield: 93%). LC-MS 424 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.78 (d, J = 1.2 Hz, 1 H), 8.50 (s, 1 H), 8.22 (d, J = 7.2 Hz, 1 H), 8.01- 8.04 (m, 1 H), 7.45 (d, J = 8.4 Hz, 1 H), 6.69 (s, 2 H), 4.54 (d, J = 3.2 Hz, 1 H), 2.92-2.97 (m, 1 H), 2.60-2.63 (m, 3 H), 1.99-2.09 (m, 1 H), 1.88-1.99 (m, 1 H).

Example 27: Preparation of A31-001 (Figure 35)

To a solution of compound A02-189 (515 mg, 1.63 mol) in DMF (5 mL) was added 2-hydroxyacetic acid (248 mg, 3.3 mmol), HATU (931 mg, 2.4 mmol), triethylamine (0.70 mL, 5.0 mmol). The reaction mixture was stirred at 80 °C for 20 h. After cooled down, the reaction mixture was evaporated and the residue is purified by Prep-HPLC to give A31-001 as a white solid (438 mg, yield: 71%). LC-MS 424 (M+H)+, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.48 (d, J = 5.2 Hz, 1 H), 8.21-8.24 (m, 1 H), 8.05-8.11 (m, 1 H), 7.66-7.79 (m, 2 H), 6.70 (s, 1 H), 4.55-4.58 (m, 1 H), 4.10 (s, 1 H), 3.14-3.19 (m, 1 H), 2.93-2.98 (m, 1 H), 2.73-2.77 (m, 1 H), 2.60-2.63 (m, 2 H), 1.90-2.07 (m, 2 H). The compound A31-001 (400 mg) was purified by Prep-Chiral-SFC (Supercritical fluid chromatography, Co-Solvent: MeOH (0.1% DEA) 35%, Column: OZ-H (4.6 x 250 mm, 5 urn)) to give product A31-001-A (158 mg) and A31-001-B (218 mg).

A31-001-A LC-MS 374 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.47-8.49 (m, 1 H), 8.21-8.24 (m, 1 H), 8.05 (d, J = 8.0 Hz, 1 H), 7.66-7.78 (m, 2 H), 6.69 (s, 2 H), 4.56-4.57 (m, 1 H), 4.09-4.10 (m, 1 H), 3.16-3.17(m, 1 H), 2.93- 2.98 (m, 1 H), 2.57-2.61 (m, 3 H), 2.07-2.08 (m, 1 H), 1.87-1.93 (m, 1 H).

A31-001-B LC-MS 374 (M+H)⁺, Purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.47-8.49 (m, 1 H), 8.21-8.24 (m, 1 H), 8.05 (d, J = 7.2 Hz,l H), 7.66-7.78 (m, 2 H), 6.69 (s, 1 H), 4.55-4.57 (m, 1 H), 4.11 (s, 1 H), 3.17 (s, 2 H), 2.93-2.98 (m, 1 H), 2.57-2.63 (m, 3 H), 2.06-2.09 (m, 1 H), 1.88-1.93 (m, 1 H).

Example 28: Preparation of A31-002 (Figure 36)

To a solution of compound A02-189 (150 mg 0.48 mmol) in DMF (5.0 mL) was added 2-(dimethylamino) acetic acid (58 mg, 0.57 mmol), HATU (271 mg, 0.71 mmol), triethylamine (0.15 mL, 1.1 mmol). After stirred at 80 °C for 20 h, the reaction mixture was purified directly by Prep-HPLC to give A31-002 as a white solid (45 mg, yield: 23%>). LC- MS 401 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.49(s, 1 H), 8.21-8.24 (m, 1 H), 8.09 (d, J = 7.6 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.66-7.71(m, 1 H), 6.04-6.05 (m, 1 H), 4.62-4.65 (m, 1 H), 3.2 (s, 2 H), 3.09-3.17 (m, 1 H), 2.69-2.79 (m, 3 H), 2.26 (s, 6 H), 2.09-2.17 (m, 1 H), 1.98-2.00 (m, 1 H).

Example 29: Preparation of A31-003 (Figure 37)

To a solution of compound A02-189 (150 mg, 0.48 mmol) in dichloromethane (10 mL) was added propionyl chloride (52 mg, 0.57 mmol), triethylamine (0.5 mL, 3.6 mmol). The mixture was stirred at room temperature for 20 h. Dichloromethane was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A31-003 as a white solid (60 mg, yield: 34%). LC-MS 372 (M+H)⁺, Purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.49 (s, 1 H), 8.21-8.24 (m, 1 H), 8.084 (d, J = 7.2 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.66-7.71 (m, 1 H), 4.61-4.64 (m, 1 H), 3.12-3.17 (m, 1 H), 2.76 (t, J = 9.6 Hz, 3 H), 2.37-2.43 (m, 2 H), 2.14-2.17 (m, 1 H), 1.95-2.03 (m, 1 H), 1.08 (t, J = 7.6 Hz, 3 H). Example 30: Preparation of A31-004 (Figure 38)

To a solution of compound A02-189 (100 mg 0.32 mmol) in dichloromethane (10 mL) was added propionyl chloride (52 mg, 0.57 mmol), triethylamine (0.5 mL, 3.6 mmol). The reaction mixture was stirred at room temperature for 20 h. Dichloromethane was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A31-004 as a white solid (66 mg, yield: 54%). LC-MS 384 (M+H)⁺, Purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.49 (s, 1 H), 8.21-8.24 (m, 1 H), 8.08 (d, J = 7.6 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.67-7.71 (m, 1 H), 4.63 (s, 1 H), 3.11-3.16 (m, 1 H), 2.71-2.77 (m, 3 H), 1.98-2.14 (m, 1 H), 1.88-1.98 (m, 2 H), 0.88 (t, J = 4.0 Hz, 4 H).

Example 31: Preparation of A31-005 (Figure 39)

To a solution of compound A02-189 (100 mg 0.32 mmol) in dichloromethane (10 mL) was added isobutyryl chloride (40 mg, 0.38 mmol), triethylamine (0.5 mL, 3.6 mmol). The reaction mixture was stirred at room temperature for 20 h. Dichloromethane was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A31-005 as a white solid (20 mg, yield: 16%). LC-MS 386 (M+H)⁺, Purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 11.88 (s, 1 H), 8.49 (s, 1 H), 8.21-8.24 (m, 1 H), 8.77 (d, J = 7.2 Hz, 1 H), 7.69-7.79 (m, 2 H), 4.630 (s, 1 H), 3.12-3.17 (m, 1 H), 2.67-2.78 (m, 4 H), 1.97-2.14 (m, 2 H), 1.09-1.17 (m, 6 H).

Example 32: Preparation of A31-006 (Figure 40)

To a solution of compound A02-189 (100 mg 0.32 mmol) in dichloromethane (10 mL) was added isobutyryl chloride (51 mg, 0.38 mmol), triethylamine (0.5 mL, 3.6 mmol). The reaction mixture was stirred at room temperature for 20 h. Dichloromethane was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A31-006 as a white solid (41 mg, yield: 31%). LC-MS 414 (M+H)⁺, Purity 100% ( UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 11.85 (s, 1 H), 8.49 (s, 1 H), 8.22-8.25 (m, 1 H), 8.08 (d, J = 7.6 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.67-7.71(m, 1 H), 4.62 (s, 1 H), 3.12-3.17 (m, 1 H), 2.73- 2.79 (m, 3 H), 2.28 (s, 2 H), 2.14-2.17 (m, 1 H), 1.97-2.00 (m, 1 H), 0.90-0.94 (m, 9 H). Example 33: Preparation of A31-007 (Figure 41)

To a solution of compound A02-189 (150 mg 0.48 mmol) in dichloromethane (10 mL) was added 2-methoxyacetyl chloride (61 mg, 0.57 mmol), triethylamine (0.5 mL, 3.6 mmol). The reaction mixture was stirred at room temperature for 20 h. Dichloromethane was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A31- 007 as a white solid (88 mg, yield: 47%). LC-MS 388 (M+H)⁺, Purity 100% ( UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 11.68 (s, 1 H), 8.49 (s, 1 H), 8.21-8.25 (m, 1 H), 8.09 (d, J = 7.6 Hz,l H), 7.67-7.79 (m, 2 H), 4.63-4.64 (m, 1 H), 4.12 (s, 2 H), 3.33 (s, 3 H), 3.14- 3.19 (s, 1 H), 2.73-2.79 (m, 3 H), 1.98-2.15 (m, 2 H). Example 34: Preparation of A31-008 (Figure 42)

To a solution of compound A02-189 (150 mg 0.48 mmol) in DMF (5.0 mL) was added 2-morpholinoacetic acid (82 mg, 0.57 mmol), HATU (271 mg, 0.71 mmol), triethylamine (0.15 mL, 1.1 mmol). The reaction mixture was stirred at 80 °C overnight. The residue was purified by Prep-HPLC to give A31-008 as a white solid (70 mg, yield: 33%). LC-MS 443 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 11.67 (s, 1 H), 8.49 (s, 1 H), 8.21-8.24 (m, 1 H), 8.08 (d, J = 7.6 Hz, 1 H), 7.66-7.79 (m, 2 H), 4.61-4.65 (m, 1 H), 3.60 (t, J = 4.4 Hz, 4 H), 3.26 (s, 2 H), 3.14 (d, J = 5.2 Hz, 1 H), 2.73-2.79 (s, 3 H), 2.49-2.50 (m, 3 H), 2.15-2.17 (m, 1 H), 1.96-2.00 (m, 1 H).

Example 35: Preparation of A25-006 (Figure 43)

To a solution of 4-chloro-6-nitroquinazoline (100 mg, 0.32 mmol), and triethylamine (95 mg, 0.95 mmol) in isopropyl alcohol (10 mL) was added 4,5,6,7-tetrahydrobenzothiazole- 2,6- diamine (67 mg, 0.32 mmol). The reaction mixture was stirred at 70 °C for 3 h. After cooled down, the reaction mixture was concentrated under reduced pressure. The residue was purified by Pre-HPLC to give the desired product A25-006 (50 mg, yield: 46%>) as yellow solid. LC-MS 343 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 9.48 (d, J = 2.4 Hz, 1 H), 8.89 (d, J = 7.2 Hz, 1 H), 8.62 (s, 1 H), 8.49 (dd, J = 9.2, 2.4 Hz, 1 H), 7.83 (d, J = 9.2 Hz, 1 H), 6.71 (s, 2 H), 4.64-4.60 (m, 1 H), 2.98 (dd, J = 15.2, 5.2Hz, 1 H), 2.69-2.59 (m, 3 H), 2.12-2.09 (m, 1 H), 1.99-1.93 (m, 1 H).

Example 36: Preparation of A02-121 (Figure 44)

To a solution of 4-chloro-6-fluoroquinazoline (200 mg, 1.1 mmol) and 1,2,3,4- tetrahydronaphthalen-2-amine (200 mg, 1.1 mmol) in isopropyl alcohol (20 mL) was added triethylamine (0.5 mL, 3.6 mmol). The resulting solution was heated to 65 °C under nitrogen overnight. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A02-121 as a white solid (33 mg, yield: 10%). LC-MS 294 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.67 (s, 1 H), 7.85-7.86 (m, 1 H), 7.47-7.52 (m, 1 H), 7.29 (s, 1 H), 7.11-7.18 (m, 4 H), 5.50 (d, J = 6.8 Hz, 1 H), 4.75-4.79 (m, 1 H), 3.33-3.38 (m, 1 H), 2.82-3.03 (m, 3 H), 2.25-2.26 (m, 1 H), 1.97-2.01 (m, 1 H).

Example 37: Preparation of A22-060 (Figure 45)

To a solution of 4-chloro-6-iodoquinazoline (986 mg, 3.4 mmol) and 1,2,3,4- tetrahydronaphthalen-2-amine (500 mg, 3.4 mmol) in isopropyl alcohol (30 mL) was added triethylamine (2.0 mL, 14 mmol). The resulting solution was heated to 90 °C under nitrogen for overnight. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A22-060 as a white solid (450 mg, yield: 33%). LC-MS 402 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, CDCls) δ 8.67 (s, 1 H), 8.02-7.94 (m, 2 H), 7.58-7.55 (m, 1 H), 7.17-7.11 (m, 4 H), 5.73- 5.70 (m, 1 H), 4.75 (m, 1 H), 3.38-3.32 (m, 1 H), 3.02-2.96 (m, 2 H), 2.89-2.82 (m, 1 H), 2.00-1.96 (m, 2 H), 2.35-2.29 (m, 2 H).

Example 38: Preparation of A22-074 (Figure 46)

To a solution of A22-060 (170 mg, 0.42 mmol) in dry DMA (5 mL) was added CuCN (195 mg, 2.1 mmol). The reaction mixture was stirred at 110 °C for 20 h under nitrogen atmosphere. The reaction mixture was diluted by water (20 mL), extracted with ethyl acetate (3 x 30 mL). The combined organic layer was washed with brine (20 mL), dried over anhydrous Na₂S0₄, filtered and concentrated. The residue was purified by Prep-HPLC to give A22-074 as a white solid (45 mg, yield: 35%). LC-MS 301(M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d₆) δ 9.00 (s, 1 H), 8.59 (s, 1 H), 8.48-8.46 (m, 1 H), 8.09-8.06 (m, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.14-7.11 (m, 4 H), 4.59-4.57 (m, 1 H), 3.21-3.16 (m, 1 H), 2.94-2.85 (m, 3 H), 2.18-2.15 (m, 1 H), 1.90-1.85 (m, 1 H).

Example 39: Preparation of A22-079 (Figure 47)

To a solution of compound A22-060 (250 mg, 0.62 mmol) in dry DMSO (6 mL) were added Cul (11.4 mg, 0.06 mmol) and sodium methanesulfmate (294 mg, 2.50 mmol). The reaction mixture was stirred at 120 °C for 2 h under microwave. The resulting mixture was quenched by water (50 mL), extracted with ethyl acetate (3 x 50 mL) and washed with water (20 mL) and brine (30 mL). The combined organic layer was dried over anhydrous sodium sulfate and filtered. The residue was purified by Prep-HPLC to give A22-079 as a white solid. (80 mg, yield: 61%). LC-MS 354 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, CDC1₃) δ 8.75 (s, 1 H), 8.44 (s, 1 H), 8.18-8.15 (m, 1 H), 8.00 (d, J = 8.8 Hz, 1 H), 4.91 (s, 1 H), 3.35-3.31 (m, 1 H), 3.14 (s, 3 H), 3.02-2.99 (m, 2 H), 2.35-2.31 (m, 1 H), 2.01-1.98 (m, 1 H).

Example 40: Preparation of A22-117 (Figure 48)

To a solution of 4-chloro-6-nitroquinazoline (80 mg, 0.38 mmol) and 1,2,3,4- tetrahydronaphthalen-2-amine (70 mg, 0.38 mmol) in isopropyl alcohol (10 mL) was added triethylamine (1 15 mg, 1.1 mmol). The resulting solution was heated to 60 °C under nitrogen for 20 h. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A22-117 as a yellow solid (40 mg, yield: 33%). LC-MS 321 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.50 (d, J = 2.4 Hz, 1 H), 8.90 (d, J = 7.6 Hz, 1 H), 8.63 (s, 1 H), 8.49 (dd, J = 9.2, 2.4 Hz, 1 H), 7.85 (d, J = 9.2 Hz, 1 H), 7.15-7.12 (m, 4 H), 4.64-4.60 (m, 1 H), 3.22- 3.17 (m, 1 H), 2.97-2.91 (m, 3 H), 2.21-2.09 (m, 1 H), 1.94-1.87 (m, 1 H).

Example 41: Preparation of A28-001 (Figure 49)

To a solution of 4-chloro-6-iodoquinazoline (500 mg, 1.7 mmol) and 1,2,3,4- tetrahydronaphthalen-1 -amine (252 mg, 1.7 mmol) in isopropyl alcohol (30 mL) was added triethylamine (0.7 mL, 5.0 mmol). The resulting solution was heated to 60 °C under nitrogen for 20 h. LC-MS analysis showed completed consumption of starting material. The mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A28-001 as a yellow solid (400 mg, yield: 60%). LC-MS 402 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.82 (d, J = 2.0 Hz, 1H), 8.62 (d, J = 8.0 Hz, 1 H), 8.53 (s, 1 H), 8.02 (d, J = 8.8 Hz, 1 H), 7.48 (d, J = 8.8 Hz, 1 H), 5.73-5.70 (m, 1 H), 2.84-2.81 (m, 2 H), 2.03-1.90 (m, 4 H).

Example 42: Preparation of A22-142 (Figure 50)

To a solution of A28-001 (150 mg, 0.37 mmol) in dry DMSO (10 mL) were added Cul (15 mg, 0.08 mmol) and sodium methanesulfmate (152 mg, 1.50 mmol). The mixture was stirred at 140 °C for 16 h. The resulting mixture was diluted by water (50 mL), extracted with ethyl acetate (3 x 50 mL). The combined organic layer was washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue is purified by Prep-HPLC to give A22-142 as a white solid. (80 mg, yield: 61%). LC-MS 354 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.13 (d, J = 8.0 Hz, 1 H), 9.06 (d, J = 2.0 Hz, 1H), 8.63 (s, 1 H), 8.20 (dd, J = 8.8 Hz, 2.0 Hz, 1 H), 7.88 (d, J = 8.8 Hz, 1 H), 7.22-7.16 (m, 4 H), 5.79-5.74 (m, 1 H), 3.31 (s, 3 H), 2.85-2.82 (m, 2 H), 2.07-1.92 (m, 4 H).

Example 43: Preparation of A02-178 (Figure 51)

To a solution of 6-fluoro-tetrahydronaphthalen-2-one (1.5 g, 9.1 mmol) and Pd/C (10%, 800 mg) in methanol (150 mL) was added a solution of ammonium formate (5.8 g, 91 mmol) in water (25 mL). The reaction mixture was stirred at 40 °C for 16 h. After cooled down to room temperature. The reaction mixture was concentrated and the resultant oil was taken up in water (80 mL). The crude product was acidified with aqueous HC1 (6 M) to pH = 4 - 5 and washed with ethyl acetate (20 mL). The aqueous phase was adjusted to pH about 10 with sodium hydroxide and extracted with ether (3 x 50 mL), The combined ether layers were dried over sodium sulfate, filtered, and concentrated to provide 6-fluoro- tetrahydronaphthalen-2-amine (1.1 g, 73%) as oil, . LC-MS 166 (M+H)⁺, purity 100% (UV 214 nm).

To a solution of 4-chloro-6-fluoroquinazoline (70 mg, 0.38 mmol) and 1,2,3,4- tetrahydronaphthalen-2-amine (64 mg, 0.38 mmol) in isopropyl alcohol (10 mL) was added triethylamine (1 16 mg, 1.1 mmol). The resulting solution was heated to 60 °C under nitrogen foe 16 h. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A02-178 as a white solid (60 mg, yield: 50%). LC-MS 312 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.48 (s, 1 H), 8.25 (dd, J = 7.2 Hz, 2.4 Hz, 1 H), 8.06 (d, J = 7.6 Hz, 2 H), 7.79-7.61 (m, 2 H), 7.17-6.94 (m, 3 H), 4.58-4.53 (m, 1 H), 3.16 (dd, J = 8.0 Hz, 4.6 Hz, 1 H), 2.94-2.79 (m, 3 H), 2.14-2.18 (m, 1H), 1.88-1.81 (m, 1 H).

Example 44: Preparation of A26-008 (Figure 52)

To a solution of 4,6-dichloroquinazoline (70 mg, 0.35 mmol) and 6-fluoro-l,2,3,4- tetrahydronaphthalen-2-amine (58 mg, 0.35 mmol) in isopropyl alcohol (10 mL) was added triethylamine (1 16 mg, 1.1 mmol). The resulting solution was heated to 60 °C under nitrogen for 16 h. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A26-008 as a white solid (50 mg, yield: 43%). LC-MS 328 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.55 (d, J = 2.4 Hz, 1 H), 8.50 (s, 1 H), 8.22 (d, J = 7.6 Hz, 2 H), 7.80 (dd, J = 8.8 Hz, 2.4 Hz, 1 H), 7.71 (d, J = 8.8 Hz, 1 H), 7.17-6.94 (m, 3 H), 4.57-4.53 (m, 1 H), 3.19-3.14 (m, 1 H), 2.94-2.82 (m, 3 H), 2.14-2.18 (m, 1 H), 1.88-1.81 (m, 1 H).

Example 45: Preparation of A28-002 (Figure 53)

To a solution of 4-chloro-6-iodoquinazoline (124 mg, 0.42 mmol) and 6-fluoro- l,2,3,4-tetrahydronaphthalen-2-amine (70 mg, 0.42 mmol) in isopropyl alcohol (10 mL) was added triethylamine (127 mg, 1.3 mmol). The resulting solution was heated to 65 °C under nitrogen for 18 h. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A28-002 as a white solid (60 mg, yield: 34%). LC-MS 420 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.80 (d, J = 2.0 Hz, 1 H), 8.50 (s, 1 H), 8.23 (d, J = 7.2 Hz, 1 H), 8.04 (dd, J = 9.2 Hz, 2.0 Hz, 1 H), 7.47 (d, J = 8.8 Hz, 1 H), 7.16 (t, J = 6.8 Hz, 1 H), 6.98-6.94 (m, 2 H), 4.62-4.58 (m, 1 H), 3.16 (dd, J = 16.0 Hz, 5.2 Hz, 1 H), 2.94-2.85 (m, 3 H), 2.14- 2.16 (m, 1 H), 1.88-1.80 (m, 1 H).

Example 46: Preparation of A23-009 (Figure 54)

To a solution of A28-002 (100 mg, 0.24 mmol) in dry DMA (6 mL) was added CuCN (64 mg, 0.72 mmol). The reaction mixture was stirred at 140 °C for 16 h under nitrogen atmosphere. The resulting mixture was poured into water (20 mL), extracted with ethyl acetate (3 x 30 mL). The combined organic layer was washed with water (20 mL), brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by Prep-HPLC to give A23-009 as a white solid (40 mg, yield: 52%). LC-MS 319 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.99 (s, 1 H), 8.60 (s, 1 H), 8.47 (d, J = 7.2 Hz, 1 H), 8.08 (dd, J = 8.4 Hz, 1.6 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.16 (t, J = 5.6 Hz, 1 H), 6.95 (d, J = 9.2 Hz, 2 H), 4.59-4.55 (m, 1 H), 3.18 (dd, J = 16.0 Hz, 4.8 Hz, 1 H), 2.95-2.91 (m, 2 H), 2.83 (dd, J = 16.0 Hz, 10.0 Hz, 1 H), 2.18-2.14 (m, 1 H), 1.90-1.83 (m, 1 H).

Example 47: Preparation of A24-006 (Figure 55)

To a solution of 4-chloro-6-methylquinazoline (75 mg, 0.42 mmol) and 6-fluoro- l,2,3,4-tetrahydronaphthalen-2-amine (70 mg, 0.42 mmol) in isopropyl alcohol (10 mL) was added triethylamine (127 mg, 1.3 mmol). The resulting solution was heated to 65 °C under nitrogen for overnight. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A24-006 as a white solid (60 mg, yield: 46%). LC-MS 420 (M+H)⁺, purity 100% (UV 214 nm); ¹HNMR (400 MHz, DMSO-d6) δ 8.43 (s, 1 H), 8.15 (s, 1 H), 7.97 (d, J = 7.2 Hz, 1 H), 7.60 (s, 2 H), 7.17-6.94 (m, 3 H), 4.62-4.58 (m, 1 H), 3.18-3.13 (m, 1 H), 2.94-2.85 (m, 3 H), 2.43 (s, 3 H),2.14-2.18 (m, 1 H), 1.88-1.82 (m, 1 H).

Example 48: Preparation of A25-003 (Figure 56)

To a solution of 4-chloro-6-nitroquinazoline (88 mg, 0.42 mmol) and 6-fluoro-l, 2, 3, 4-tetrahydronaphthalen-2-amine (70 mg, 0.42 mmol) in isopropyl alcohol (10 mL) was added triethylamine (127 mg, 1.3 mmol). The resulting solution was heated to 65 °C under nitrogen overnight. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A25-003 as a white solid (70 mg, yield: 49%). LC-MS 420 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.49 (d, J = 2.4 Hz, 1 H), 8.89 (d, J = 6.8 Hz, 1 H), 8.63 (s, 1 H), 8.49 (dd, J = 7.2 Hz, 2.4 Hz, 1 H), 7.85 (d, J = 9.2 Hz, 1 H), 7.18-6.95 (m, 3 H), 4.62-4.58 (m, 1 H), 3.19 (dd, J = 16.0 Hz, 4.8 Hz, 1H), 2.94-2.88 (m, 3 H), 2.14-2.18 (m, 1 H), 1.94-1.84 (m, 1 H). Example 49: Preparation of A02-184 (Figure 57)

To a solution of tetrahydronaphthalen-2-one (8.0 g, 54.8 mmol) and Pd/C (10%, 800 mg) in methanol (300 mL) was added a solution of ammonium formate (34.5 g, 0.55 mmol) in water (50 mL). The reaction mixture was stirred at 40 °C for 16 h. After cooled down to room temperature the reaction mixture was concentrated. The resultant oil was taken up in water (150 mL). The crude product was acidified with HC1 (6 M) to pH = 4 - 5 and washed with ethyl acetate. The aqueous phase was adjusted to pH about 10 with sodium hydroxide (5 M), and extracted with ether (2 x 50 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated to provide tetrahydronaphthalen-2-amine (5.0 g, yield: 50%) as oil. LC-MS 148 (M+H)⁺, purity 90% (UV 214 nm).

Trifluoroacetic anhydride (5.7 mL, 41.0 mmol) was added to a solution of compound 2 (5.0 g, 27.3 mmol) in dichloromethane (100 mL) in the presence of triethylamine (7.6 mL, 54.6 mmol) and DMAP (133 mg, 1.1 mmol) in ice-bath. The reaction mixture was stirred at room temperature overnight, and then washed with aqueous HC1 (0.5 M, 10 mL) and water (25 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (using petroleum ether : ethyl acetate = 9:1 - 7:3) to give compound 6 (6.0 g, yield: 90%>). LC-MS 244 (M+H)⁺, purity 95% (UV 214 nm).

To a solution of compound 3 (3.0 g, 12.3 mmol) in nitromethane (45 mL) at -5 °C was added a solution of sulfuric acid (14 mL), nitric acid (1.0 mL) and water (2.50 mL) dropwise over 30 minute. The reaction mixture was stirred at 0 °C for 2 h. The reaction mixture was poured into water (50 mL) and extracted with ethyl acetate (3 x 60 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated to give the compound mixture 7 as yellow oil (3.3 g, yield: 95%>).

A mixture of compound 7 (2.0 g, 6.9 mmol) and Pd/C (10%, 300 mg) in MeOH (30 mL) was stirred at 20 °C under hydrogen atmosphere (1 atm) for 16 h. The reaction mixture was filtered and the filtrate was concentrated with rotary evaporator. The residue was purified by column chromatography to give compound 8a (350 mg, yield: 19%), compound 8b (550 mg, yield: 31%), a mixture of 8c and 8d (950 mg of mixture 8c and 8d, yield: 53%).

To a solution of compound 8a (300 mg, 1.16 moL) and ammonium hydroxide (5 mL) in ethanol (15 mL) was added potassium carbonate (1.6 g, 11.6 mmol). The reaction mixture was stirred at 90 °C for 5 h. After cooled down, the reactin mixture was concentrated under reduced pressure. The crude product was taken up in water (30 mL), and extracted with ethyl acetate (2 x 50 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated in vacuo to give the compound 9a as yellow oil (150 mg, yield: 80%).

To a solution of compound 8b (300 mg, 1.16 moL) and ammonium hydroxide (5 mL) in ethanol (15 mL) was added potassium carbonate (1.6 g, 11.6 mmol). The reaction mixture was stirred at 90 °C for 5 h. After cooled down, the reactin mixture was concentrated under reduced pressure. The crude product was taken up in water (30 mL), and extracted with ethyl acetate (2 x 50 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated in vacuo to give the compound 9b as yellow oil (145 mg, yield: 77%).

To a solution of the mixture of 8c and 8d (600 mg, 2.32 mmol) and ammonium hydroxide (10 mL) in ethanol was added potassium carbonate (3.2 g, 23.2 mmol). The reaction mixture was stirred at 90 °C for 5 h. After cooled down, the reactin mixture was concentrated under reduced pressure. The crude product was taken up in water (30 mL), and extracted with ethyl acetate (2 x 50 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by Prep-HPLC to give the desired compound 9c (132 mg, yield: 35%>) as white solid and compound 9d (113 mg, yield: 30%>) as white solid.

To a solution of 4-chloro-6-fluoroquinazoline (95 mg, 0.52 mmol) and compound 9a (85 mg, 0.52 mmol) in isopropyl alcohol (10 mL) was added triethylamine (157 mg, 1.6 mmol). The resulting solution was heated to 65 °C under nitrogen for overnight. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A02-184 as a white solid (50 mg, yield: 31%). LC-MS 309 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.48 (s, 1 H), 8.26 (dd, J = 10.0 Hz, 2.4 Hz, 1 H), 8.14 (d, J = 7.6 Hz, 1 H), 7.77 (dd, J = 9.2 Hz, 5.6 Hz, 1 H), 7.69 (td, J = 9.2 Hz, 2.8 Hz, 1 H), 6.83 (t, J = 7.6 Hz, 1 H), 6.46 (d, J = 7.6 Hz, 1 H), 6.34 (d, J = 7.6 Hz, 1 H), 4.76 (s, 2 H) 4.63-4.59 (m, 1 H), 2.96 (dd, J = 16.4 Hz, 6.0 Hz, 1 H), 2.79-2.85 (m, 2 H), 2.38 (dd, J = 16.4 Hz, 10.0 Hz, 1 H), 2.05-2.09 (m, 1 H), 1.87-1.79 (m, 1 H).

Example 50: Preparation of compound A02-185 (Figure 58)

To a solution of 4-chloro-6-fluoroquinazoline (95 mg, 0.52 mmol) and compound 9b (80 mg, 0.49 mmol) in isopropyl alcohol (10 mL) was added triethylamine (157 mg, 1.6 mmol). The resulting solution was heated to 65 °C under nitrogen for overnight. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A02-185 as a white solid (50 mg, yield: 36%). LC-MS 309 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.48 (s, 1 H), 8.25 (dd, J = 10.0 Hz, 2.8 Hz, 1 H), 8.03 (d, J = 6.8 Hz, 1 H), 7.77 (dd, J = 9.2 Hz, 6.0 Hz, 1 H), 7.69 (td, J = 9.2 Hz, 2.8 Hz, 1 H), 6.83 (t, J = 8.0 Hz, 1 H), 6.47 (d, J = 8.0 Hz, 1 H), 6.33 (d, J = 7.62 Hz, 1 H), 4.79 (s, 2 H) 4.63-4.59 (m, 1 H), 3.03 (dd, J = 15.6 Hz, 4.0 Hz, 1H), 2.81 (dd, J = 15.6 Hz, 10.8 Hz, 1 H), 2.66 (dd, J = 17.2 Hz, 4.0 Hz, 1 H), 2.52-2.43 (m, 1 H), 2.24-2.20 (m, 1 H) 1.85-1.78 (m, 1 H).

Example 51: Preparation of compound A02-186 (Figure 59)

To a solution of 4-chloro-6-fluoroquinazoline (56 mg, 0.3 mmol) and 9c (50 mg, 0.3 mmol) in DMF (4 mL) was added potassium carbonate (128 mg, 0.93 mmol). The resulting solution was heated to 80 °C under nitrogen for overnight. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and poured into water (20 mL), extracted with ethyl acetate (2 x 30 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by Prep-HPLC to give A02-186 as a white solid (70 mg, yield: 74%). LC-MS 309 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.47 (s, 1 H), 8.24 (dd, J = 10.0 Hz, 2.4 Hz, 1 H), 8.01 (d, J = 7.2 Hz, 1 H), 7.76 (dd, J = 8.8 Hz, 6.0 Hz, 1 H), 7.67 (td, J = 8.8 Hz, 2.8 Hz, 1 H), 6.75 (d, J = 8.0 Hz, 1 H), 6.39-6.31 (m, 2 H), 4.80 (s, 2 H) 4.49- 4.45 (m, 1 H), 2.99 (dd, J = 15.6 Hz, 5.2 Hz, 1 H), 2.80-2.67 (m, 3 H), 2.13-2.08 (m, 1 H), 1.82-1.72 (m, 1 H). Example 52: Preparation of compound A02-187 (Figure 60)

To a solution of 4-chloro-6-fluoroquinazoline (56 mg, 0.3 mmol) and compound 9d (50 mg, 0.3 mmol) in DMF (4 mL) was added potassium carbonate (128 mg, 0.93 mmol). The resulting solution was heated to 80 °C under nitrogen for overnight. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and poured into water (20 mL), extracted with ethyl acetate (2 x 30 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by Prep-HPLC to give A02-186 as a white solid (70 mg, yield: 74%). LC-MS 309 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.47 (s, 1 H), 8.24 (d, J = 10.0 Hz, 1 H), 8.01 (d, J = 7.2 Hz, 1 H), 7.76 (dd, J = 8.8 Hz, 6.0 Hz, 1 H), 7.67 (td, J = 8.8 Hz, 2.4 Hz, 1 H), 6.77 (d, J = 8.0 Hz, 1 H), 6.38 (dd, J = 8.4 Hz, 2.0 Hz, 1 H), 6.31 (s, 1 H), 4.78 (s, 2 H) 4.49-4.45 (m, 1 H), 2.99 (dd, J = 16.0 Hz, 4.8 Hz, 1 H), 2.80-2.67 (m, 3 H), 2.13-2.10 (m, 1 H), 1.79-1.72 (m, 1 H).

Example 53: Preparation of A23-016 (Figure 61)

To a solution of 4-chloro-6-iodoquinazoline (144 mg, 0.49 mmol) and 1, 2, 3, 4 - tetrahydronaphthalene-2,6-diamine (80 mg, 0.49 mmol) in DMF (10 mL) was added potassium carbonate (205 mg, 1.48 mmol). The resulting solution was heated to 90 °C under nitrogen for overnight. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and poured into water (20 mL), extracted with ethyl acetate (2 x 30 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated in vacuo to give the crude product compound A23-016-1 (62 mg yield: 30%). LC-MS 417 (M+H)⁺, purity 70% (UV 214 nm).

To a solution of compound A23-016-1 (62 mg, crude, 0.15 mmol) and Boc₂0 (65 mg, 0.3 mmol) in THF (10 mL) was added triethylamine (46 mg, 0.45 mmol). The resulting solution was heated to 60 °C under nitrogen for 2 h. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and excess THF was removed by rotary evaporation. The residue was purified by silica gel column chromatography (using petroleum ether : ethyl acetate = 7:3 - 1 : 1) to give compound A23- 016-2 as a white solid (74 mg, yield: 97%). LC-MS 517 (M+H)⁺, purity 90% (UV 214 nm).

The compound 3 (74 mg, crude, 0.14 mmol), zinc cyanide (50 mg, 0.43 mmoL), Zn (8 mg, 0.1 mmol), Pd(dppf)Cl₂ (15 mg, 0.02 mmol) was dissolved in DMF (10 mL). The resulting solution was heated to 140 °C in microwave for 2 h. The reaction mixture was poured into water and extracted with ethyl acetate (2 x 50 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated. The residue was purified by Prep-HPLC to give A23-016 as a white solid (30 mg, yield: 66%). LC-MS 316 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.00 (s, 1 H), 8.58 (s, 1 H), 8.43 (d, J = 2.4 Hz, 1 H), 8.08 (dd, J = 8.8 Hz, 2.2 Hz, 1 H), 7.78 (d, J = 8.4 Hz, 1 H), 6.75 (d, J = 8.0 Hz, 1 H), 6.37 (d, J = 8.0 Hz, 1 H), 6.34 (s, 1 H), 4.81 (s, 2 H) 4.51-4.49 (m, 1 H), 2.99 (dd, J = 15.6 Hz, 4.8 Hz, 1 H), 2.80-2.70 (m, 3 H), 2.13-2.08 (m, 1 H), 1.82-1.77 (m, 1 H).

Example 54: Preparation of A23-025 (Figure 62)

To a solution of 4-chloro-6-iodoquinazoline (144 mg, 0.49 mmol) and 1, 2, 3, 4 - tetrahydronaphthalene-l,6-diamine 9b (80 mg, 0.49 mmol) in DMF (10 mL) was added potassium carbonate (205 mg, 1.48 mmol). The resulting solution was heated to 90 °C under nitrogen for 16 h. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and poured into water (20 mL), extracted with ethyl acetate (2 x 50 mL). The organic phase was dried over sodium sulfate, filtered, and the filtrate was evaporated in vacuo to give the crude product A23-025-1 (85 mg, yield: 41%). LC-MS 417 (M+H)⁺, purity 90% (UV 214 nm).

To a solution of compound A23-025-1 (85 mg, crude, 0.2 mmol) and Boc₂0 (89 mg, 0.4 mmol) in THF (10 mL) was added triethylamine (100 mg, 1.0 mmol). The resulting solution was heated to 60 °C under nitrogen for 8 h. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and excess THF was removed by rotary evaporation. The residue was purified by silica gel column chromatography (using petroleum ether : ethyl acetate = 2: 1) to give compound A23-025-2 as a white solid (90 mg, yield: 35%). LC-MS 517 (M+H)⁺, purity 90% (UV 214 nm).

The compound A23-025-2 (90 mg, crude, 0.17 mmol), zinc cyanide (61 mg, 0.52 mmoL), Zn (8 mg, 0.1 mmol), Pd(dppf)Cl₂ (15 mg, 0.02 mmol) was dissolved in DMF (10 mL). The resulting solution was heated to 140 °C under microwave for 2 h. The reaction mixture was poured into water and extracted with ethyl acetate (2 x 50 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated. The residue was purified by Prep-HPLC to give A23-025 as a white solid (25 mg, yield: 46%>). LC-MS 316 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.02 (d, J = 2.4 Hz, 1 H), 8.59 (s, 1 H), 8.48 (d, J = 7.2 Hz, 1 H), 8.09 (dd, J = 8.8 Hz, 2.0 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 6.84 (t, J = 7.6 Hz, 1 H), 6.48 (d, J = 7.6 Hz, 1 H), 6.34 (d, J = 7.2 Hz, 1 H), 5.0 (s, 1 H), 4.50-4.46 (m, 1 H), 3.04 (dd, J = 16.4 Hz, 4.4 Hz, 1 H), 2.82-2.77 (m, 3 H), 2.24-2.21 (m, 1 H),l .86-1.81 (m, 1 H).

Example 55: Preparation of A23-026 (Figure 63)

To a solution of 4-chloro-6-iodoquinazoline (144 mg, 0.49 mmol) and 1, 2, 3, 4 - tetrahydronaphthalene-2,7-diamine 9d (80 mg, 0.49 mmol) in DMF (10 mL) was added potassium carbonate (205 mg, 1.48 mmol). The resulting solution was heated to 90 °C under nitrogen for 16 h. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and poured into water (20 mL), extracted with ethyl acetate (2 x 100 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated in vacuo to give the crude product A23-026-l(85 mg yield: 41%). LC-MS 417 (M+H)⁺, purity 90% (UV 214 nm).

To a solution of compound A23-026-1 (85 mg, crude, 0.2 mmol) and Boc₂0 (89 mg, 0.4 mmol) in THF (10 mL) was added triethylamine (100 mg, 1.0 mmol). The resulting solution was heated to 70 °C under nitrogen for 3 h. LC-MS analysis showed completed consumption of starting material. The reaction mixture was cooled down and excess THF was removed by rotary evaporation. The residue was purified by silica gel column chromatography (using petroleum ether : ethyl acetate = 2: 1) to give compound A23-026-2 as a white solid (80 mg, yield: 31%). LC-MS 517 (M+H)⁺, purity 90% (UV 214 nm).

The compound A23-026-2 (80 mg, 0.15 mmol), zinc cyanide (54 mg, 0.46 mmoL), Zn (8 mg, 0.1 mmol), Pd(dppf)Cl₂ (15 mg, 0.02 mmol) was dissolved in DMF (10 mL). The resulting solution was heated to 140 °C under microwave for 2 h. The reaction mixture was poured into water and extracted with ethyl acetate (2 x 30 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated. The residue was purified by Prep-HPLC to give A23-026 as a white solid (20 mg, yield: 42%). LC-MS 316 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.0 (s, 1 H), 8.59 (s, 1 H), 8.43 (d, J = 7.2 Hz, 1 H), 8.08 (dd, J = 8.8 Hz, 1.6 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 6.78 (d, J = 8.0 Hz, 1 H), 6.38 (dd, J = 8.4 Hz, 2.4 Hz, 1 H), 6.31 (s, 1 H), 4.79 (s, 2 H), 4.54- 4.48 (m, 1 H), 2.99 (dd, J = 16.0 Hz, 5.2 Hz, 1 H), 2.78-2.73 (m, 3 H), 2.17-2.13 (m, 1 H), 1.81-1.75 (m, 1 H).

Example 56: Preparation of A30-001 (Figure 64)

To a solution of compound 23-016 (45 mg, 0.14 mol) in DMF (4 mL) was added 2- hydroxyacetic acid (90 mg, 1.4 mmol), HATU (82 mg, 0.2 mmol), triethylamine (30 mg, 0.28 mmol). After stirred at 60 °C for 16 h, the reaction mixture was poured into water (20 mL) and extracted with ethyl acetate (4 x 50 mL). The organic layer washed with water (25 mL) and brine (25 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated. The residue was purified by Prep-HPLC to give A30-001 as a white solid (9 mg, yield: 16%). LC-MS 374 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.80 (s, 1 H), 8.58 (s, 1 H), 8.02 (dd, J = 8.4 Hz, 2.0 Hz, 1 H), 7.82 (d, J = 8.4 Hz, 1 H), 7.43 (s, 1 H), 7.37 (dd, J = 8.4 Hz, 2.0 Hz, 1 H), 7.10 (d, J = 8.4 Hz, 1 H), 4.70-4.66 (m, 1 H), 4.12 (s, 2 H), 3.26 (dd, J = 16.0 Hz, 4.4 Hz, 1 H), 3.04-2.88 (m, 3H), 2.31-2.28 (m, 1 H), 1.98-1.93 (m, 1 H).

Example 57: Preparation of A30-003 (Figure 65)

To a suspension of potassium tert-butanolate (5.37 g, 48 mmol) in dry ethyl ether (100 mL) at 0 °C was added 3,4-dihydronaphthalen-l(2H)-one (5.74 g, 40 mmol) in dropwise. The reaction mixture was stirred at 0 °C for 30 min. Then the resulting mixture was added t-butyl nitrite (6.18 g, 60 mmol) slowly and the resulting mixture was stirred at room temperature for 1 h. The reaction mixture was poured into cool water (300 mL) and extracted with ethyl acetate (1.0 L). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (using petroleum ether : ethyl acetate = 4: 1 - 7:3) to give the desired compound 10 (two isomers) (6.0 g, yield: 86%>) as a pale green solid. LC-MS 176 (M+H)⁺, purity 98% (UV 214 nm).

To a solution of compound 10 (4.0 g, 22.8 mmol) in a mixture of MeOH (100 mL) and AcOH (2.0 mL) were added Pd/C (400 mg, 10%) and di-tert-butyl dicarbonate (7.4 g, 34.3 mmol). The reaction mixture was stirred at room temperature for 20 h under hydrogen atmosphere. The resulting mixture was filtered off and the filtrate was concentrated and purified by silica gel column chromatography (using petroleum ether : ethyl acetate = 9:1 - 4: 1) to give the desired compound 11 (3.6 g, yield: 60%). LC-MS 286.0 (M+Na)⁺, purity 88% (UV 214 nm).

To a solution compound 11 (526 mg, 2.0 mmol) in dichloromethane was added a solution of HC1 in 1,4-dioxane (4 M, 10 mL). After stirred at room temperature for 24 h, excess solvent was removed by rotary evaporator to give the crude compound 12 (450 mg) as a light red solid, which was directly used for the next step reaction. LC-MS 164 (M+H)⁺, purity 50% (UV 214 nm).

To a solution of compound 12 (364 mg, purity 50%>) and 4-chloro-6- fluoroquinazoline (364 mg, 2.0 mmol) in isopropyl alcohol (20 mL) was added triethylamine (1.0 mL, 7.2 mmol). The resulting solution was heated to 80 °C under nitrogen for 2 h. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A30-003 as a white solid (200 mg, yield: 32%). LC-MS 374 (M+H)⁺, purity 98% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.46-8.44 (m, 1 H), 8.26-8.22 (m, 1 H), 8.03-8.01 (m, 1 H), 7.78-7.68 (m, 2 H), 7.54-7.52 (m, 1 H), 7.23-7.11 (m, 3 H), 5.51-5.49 (m, 1 H), 4.84-4.80 (m, 1 H), 4.48-4.46 (m, 1 H), 2.93-2.87 (m, 2 H), 2.15-2.13 (m, 1 H), 1.85-1.82 (m, 1 H). Example 58: Preparation of A30-007 (Figure 66)

To a solution A30-007 (154 mg, 0.5 mmol) in dichloromethane (20 mL) was slowly added DAST (161 mg, 1.0 mmol) at -78 °C. The reaction mixture was warmed up to room temperature and stirred at this temperature for 2 h. The reaction mixture was quenched with saturated solution of sodium bicarbonate (20 mL) and extracted with dichloromethane (2 x 50 mL). The organic layer was concentrated by rotary evaporation. The residue was purified by Prep-HPLC to give A30-007 as a white solid (35 mg, yield: 22%). LC-MS 312 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.49 (s, 1 H), 8.45- 8.42 (m, 1 H), 8.30-8.27 (m, 1 H), 7.82-7.68 (m, 2 H), 7.47-7.37 (m, 2 H), 7.30-7.26 (m, 2 H), 5.79-5.65 (m, 1 H), 4.74-4.64 (m, 1 H), 3.02-2.97 (m, 2 H), 2.30-2.25 (m, 1 H), 2.07- 1.98 (m, 1 H).

Example 59: Preparation of A30-002 (Figure 67)

To a solution of 2-amino-l,2,3,4-tetrahydronaphthalen-l-ol hydrochlorid 12 (580 mg, 2.0 mmol) and 4-chloro-6-iodoquinazoline (580 mg, 2.0 mmol) in isopropyl alcohol (20 mL) was added triethylamine (1.5 mL, 10.8 mmol). The resulting solution was heated to 60 °C under nitrogen for 16 h. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation. The residue was purified by Prep- HPLC to give A30-002 as a yellow solid (400 mg, yield: 48%). LC-MS 418 (M+H)⁺, purity 96% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.81 (s, 1 H), 8.48 (s, 1 H), 8.22-8.20 (m, 1 H), 8.04-8.01 (m, 1 H), 7.54-7.45 (m, 2 H), 7.22-7.10 (m, 3 H), 5.52-5.50 (m, 1 H), 4.85-4.81 (m, 1 H), 4.47-4.45 (m, 1 H), 2.93-2.83 (m, 2 H), 2.16-2.12 (m, 1 H), 1.84-1.79 (m, 1 H).

Example 60: Preparation A30-006 (Figure 68)

To a solution A30-002 (208 mg, 0.5 mmol) in dichloromethane (10 mL) was slowly added DAST (161 mg, 1.0 mmol) at -78 °C. The mixture was warmed up to room temperature and stirred at this temperature for 2 h. The reaction mixture was quenched with a saturated solution of sodium bicarbonate (20 mL) and extracted with dichloromethane (2 x 50 mL). The organic layer was concentrated by rotary evaporation and the residue was purified by pre-HPLC to give compound 13 as a white solid (250 mg, yield: 22%). LC-MS 420 (M+H)⁺, purity 66% (UV 214 nm).

To a solution of compound 13 (250 mg, 0.6 mmol) in dry DMA (4.0 mL) was added copper (I) cyanide (184 mg, 2.0 mmol). The mixture was stirred at 100 °C for 6 h under nitrogen atmosphere. The resulting mixture was poured into water (25 mL), extracted with ethyl acetate (3 x 50 mL). The organic phase was washed with water (2 x 10 mL) and brine (2 x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated by rotary evaporation. The residue was purified by Prep-HPLC to give A30-006 as a white solid (30 mg, yield: 19%). LC-MS 319 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.20 (s, 1 H), 8.70-8.68 (m, 1 H), 8.61 (s, 1 H), 8.13-8.11 (m, 1 H), 7.84-7.82 (m, 1 H), 7.46-7.38 (m, 2 H), 7.30-7.26 (m, 2 H), 5.79-5.65 (m, 1 H), 4.78-4.67 (m, 1 H), 3.04-3.00 (m, 2 H), 2.32-2.25 (m, 1 H), 2.07-2.00 (m, 1 H).

Example 61: Preparation A30-005 (Figure 69)

To a solution of N-(l-fluoro- 1,2,3, 4-tetrahydronaphthalen-2-yl)-6-iodoquinazolin-4- amine (compound 13, 150 mg, crude) in dry DMSO (4.0 mL) were added cuprous iodide (11.4 mg, 0.06 mmol) and sodium methanesulfmate (118 mg, 1.0 mmol). The reaction mixture was stirred at 110 °C for 5 h under nitrogen atmosphere. The resulting mixture was poured into water (25 mL), extracted with ethyl acetate (3 x 50 mL). The organic phase was washed with water (2 x 10 mL) and brine (2 x 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated by rotary evaporation. The residue was purified by Prep- HPLC to give A30-005 as a yellow solid (38 mg, yield: 34%). LC-MS 372 (M+H)⁺, purity 97% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.25 (s, 1 H), 9.00-8.98 (m, 1 H), 8.62 (s, 1 H), 8.24-8.22 (m, 1 H), 7.92-7.90 (m, 1 H), 7.47-7.38 (m, 2 H), 7.30-7.26 (m, 2 H), 5.79-5.66 (m, 1 H), 4.82-4.73 (m, 1 H), 3.32 (s, 3 H), 3.04-3.00 (m, 2 H), 2.34-2.29 (m, 1 H), 2.02-2.00 (m, 1 H). Example 62: Preparation of compounds A29-002B, A29-015A, A29-015B and A29- 0

To a solution of A23-001 (500 mg, 1.66 mmol) in DMF (5.0 mL) were added 3- hydroxy-3-methylbutanoic acid (392 mg, 3.32 mmoL), HATU (1.26 g, 3.32 mmol) and triethylamine (0.72 mL, 4.98 mmol). The reaction mixture was stirred at room temprature for 9 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give SP-0010462-132-A (200 mg) and SP-0010462-132-B (150 mg).

The compound SP-0010462-132-A (200 mg) was purified by Prep-Chiral-SFC (supercritical fluid chromatography, Co-Solvent: MeOH, Column: OZ-H (4.6 x 250 mm, 5 urn)) to give product A29-002B (50 mg) and A29-015A (56 mg).

A29-002B LC-MS 344 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.85 (s, 1 H), 8.98 (s, 1 H), 8.67 (d, J = 6.0 Hz, 1 H), 8.62 (s, 1 H), 8.07 (d, J = 8.4 Hz, 1 H), 7.79 (d, J = 8.4 Hz, 1 H), 7.56 (s, 1 H), 7.32 (d, J = 6.8 Hz, 1 H), 7.17 (d, J = 7.6 Hz, 1 H), 5.03-5.00 (m, 1 H), 3.39-3.30 (m, 2 H), 3.05-2.96 (m, 2 H), 2.02 (s, 3 H).

A29-015A LC-MS 344 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.86 (s, 1H), 8.98 (s, 1 H), 8.67 (d, J = 6.8 Hz, 1 H), 8.62 (s, 1 H), 8.07 (d, J = 8.8 Hz, 1 H), 7.79 (d, J = 8.4 Hz, 1 H), 7.56 (s, 1 H), 7.32 (d, J = 8.8 Hz, 1 H), 7.17 (d, J = 8.4 Hz, 1 H), 5.03-5.00 (m, 1 H), 3.39-3.30 (m, 2 H), 3.05-2.96 (m, 2 H), 2.02 (s, 3 H).

The compound SP-0010462-132-B (150 mg) was purified by Prep-Chiral-SFC (supercritical fluid chromatography, Co-Solvent: MeOH, Column: OZ-H (4.6 x 250 mm, 5 urn)) to give product A29-015B (30 mg) and A29-002A (25 mg).

A29-015B LC-MS 402 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.77 (s, 1 H), 8.98 (s, 1 H), 8.66 (d, J = 6.4 Hz, 1 H), 8.62 (s, 1 H), 8.07 (d, J = 8.4 Hz, 1 H), 7.79 (d, J = 8.4 Hz, 1 H), 7.59 (s, 1 H), 7.33 (d, J = 8.4 Hz, 1 H), 7.18 (d, J = 8.4 Hz, 1 H), 5.04-5.00 (m, 1 H), 4.75 (s, 1 H), 3.38-3.30 (m, 2 H), 3.05-2.95 (m, 2 H), 2.40 (s, 2 H), 1.22 (s, 6 H). A29-002A LC-MS 402 (M+H)⁺, purity 98% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.78 (s, 1 H), 8.97 (s, 1 H), 8.66 (d, J = 6.8 Hz, 1 H), 8.62 (s, 1 H), 8.07 (d, J = 8.4 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.59 (s, 1 H), 7.41 (d, J = 6.4 Hz, 1 H), 7.33 (d, J = 8.0 Hz, 1 H), 7.18 ( d, J = 8.0 Hz, 1 H), 5.04-4.99 (m, 1 H), 4.76 (s, 1 H), 3.36-3.30 (m, 2 H), 3.05-2.95 (m, 2 H), 2.40 (s, 2 H), 1.22 (s, 6 H).

Notes: (1) the absolute stereochemistry for the chiral center on the 5-membered ring of A29-002B, A29-015A, A29-015B and A29-002A was assigned arbitrarily. (2) the compound SP-0010462-132-A was a by-product of the coupling reaction.

Example 63: Preparation of compound A29-016

A02-171 A29-016

To a solution of compound A02-171 (100 mg, 0.340 mmol) in CHCI₃ (6 mL) cooled in an ice-bath were added 2-methoxyethanesulfonyl chloride (270 mg, 1.70 mmol), triethylamine (360 mg, 3.40 mmol) and DMAP (4 mg, 0.034 mmol). The reaction mixture was stirred at room temperature for 2 d. Water (0.1 mL) was added to quench the reaction. The resulting mixture was stirred for additional 10 min and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give A29-016 (15 mg, yield: 11%). LC- MS 417 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, MeOD-d4) δ 8.50 (s, 1 H), 8.03 (dd, J = 9.6 Hz, 2.8 Hz, 1 H), 7.78 (dd, J = 9.2 Hz, 5.2 Hz, 1 H), 7.62 (td, J = 8.4 Hz, 2.8 Hz, 1 H), 7.24-7.21 (m, 2 H), 7.10 (dd, J = 8.0 Hz, 2.0 Hz, 1 H), 5.16-5.12 (m, 1 H), 3.78 (t, J = 6.0 Hz, 2 H), 3.49-3.41 (m, 2 H), 3.32-3.29 (m, 5 H), 3.12-3.04 (m, 2 H).

Example 64: Preparation of compound A29-017

A02-171 A29-017

To a solution of compound A02-171 (80 mg, 0.272 mmol) in DCM (10 mL) cooled in an ice-bath were added acetic anhydride (33 mg, 0.327 mmol), triethylamine (410 mg, 4.08 mmol) and DMAP (4 mg, 0.033 mmol). The reaction mixture was stirred at room temperature for 2 h, and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give A29-017 (55 mg, yield: 60%). LC-MS 337 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.86 (s, 1 H), 8.52 (s, 1 H), 8.28-8.22 (m, 2 H), 7.77 (dd, J = 9.2 Hz, 5.2 Hz, 1 H), 7.68 (td, J = 8.4 Hz, 2.8 Hz, 1 H), 7.56 (s, 1 H), 7.32 (d, J = 8.0 Hz, 1 H), 7.16 (d, J = 8.0 Hz, 1 H), 5.03-4.98 (m, 1 H), 3.38-3.30 (m, 2 H), 3.00 (td, J = 16.8 Hz, 6.4 Hz, 2 H), 2.03 (s, 3 H).

Example 65: Preparation of compound A29-018

A02-171 A29-018

To a solution of compound A02-171 (80 mg, 0.272 mmol) in DMF (3 mL) were added 2-hydroxyacetic acid (25 mg, 0.327 mmol), HATU (134 mg, 0.354 mmol), triethylamine (275 mg, 2.72 mmol). The mixture was stirred at 40 °C for 5 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (2 x 20 mL). The organic layer was washed with water (10 mL) and brine (10 mL), dried over Na₂S0₄, filtered, and concentrated. The residue was purified by Prep-HPLC to give A29-018 as a white solid (30 mg, yield: 31%). LC-MS 353 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.57 (s, 1 H), 8.52 (s, 1 H), 8.28 (d, J = 6.4 Hz, 1 H), 8.23 (dd, J = 10.4 Hz, 2.4 Hz, 1 H), 7.77 (dd, J = 8.8 Hz, 5.6 Hz, 1 H), 7.70-7.66 (m, 2 H), 7.46 (d, J = 7.6 Hz, 1 H), 7.19 (d, J = 8.0 Hz, 1 H), 5.66 (t, J = 5.6 Hz, 1 H), 5.03-4.98 (m, 1 H), 3.98 (d, J = 6.0 Hz, 2 H), 3.38-3.01 (m, 2 H), 3.01 (dt, J = 16.4 Hz, 6.0 Hz, 2 H).

Example 66: Preparation of A29-019

A29-019

To a solution of A02-171 (100 mg, 0.34 mmol) in DMF (2.0 mL) were added 2- (diethylamino) acetic acid hydrochloride (114 mg, 0.68 mmoL), HATU ( 387 mg, 1.02 mmol) and triethylamine (0.24 mL, 1.70 mmol). The reaction mixture was stirred at 50 °C for 9 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-019 as a white solid (30 mg, yield: 21%). LC-MS 408 (M+H) ⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.56 (s, 1 H), 8.51 (s, 1 H), 8.26 (d, J = 6.8 Hz, 1 H), 8.22 (d, J = 10.4 Hz, 1 H), 7.78-7.75 (m, 1 H), 7.67 (t, J = 8.8 Hz, 1 H), 7.59 (s, 1 H), 7.41 (d, J = 8.0 Hz, 1 H), 7.18 (d, J = 8.4 Hz, 1 H), 5.03-4.99 (m, 1 H), 3.38-3.30 (m, 2 H), 3.12 (s, 2 H), 3.05-2.95 (m, 2 H), 2.59 (d-d, Ji =7.2 Hz, J₂ = 7.2 Hz, 4 H), 1.02 (t, J = 7.2 Hz, 6 H).

Example 67: Preparation of compound A29-020

To a solution of A23-001 (100 mg, 0.33 mmol) in DMF (2.0 mL) were added 2- (diethylamino)acetic acid hydrochloride (110 mg, 0.66 mmoL), HATU (380 mg, 1.00 mmol) and triethylamine (0.24 mL, 1.65 mmol). The reaction mixture was stirred at 50 °C for 9 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-020 as a white solid (20 mg, yield: 15%). LC-MS 415 (M+H) ⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.56 (s, 1 H), 8.98 (s, 1 H), 8.66 (d, J = 6.0 Hz, 1 H), 8.63 (s, 1 H), 8.07 (d, J = 4.0 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.60 (s, 1 H), 7.42 (d, J = 8.0 Hz, 1 H), 7.19 (d, J = 8.0 Hz, 1 H), 5.03-4.99 (m, 1 H), ), 3.12 (s, 2 H), 3.41-3.39 (m, 2 H), 3.06-2.98 (m, 2 H), 2.62-2.50 (m, 4 H), 1.02 (t, J = 6.8 Hz, 6 H).

Example 68: Preparation of compound A29-021

A23-001 50 °C, 10 h A29-021

To a solution of A23-001 (50 mg, 0.17 mmol) in DMF (2.0 mL) were added phenyl 5-tert- butylisoxazol-3-ylcarbamate (65 mg, 0.25 mmoL) and pyridine (0.04 mL, 0.51 mmol). The reaction mixture was stirred at 50 °C for 10 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-021 as a white solid (20 mg, yield: 25 %>). LC-MS 468 (M+H) ⁺, purity 99% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1 H), 8.99 (s, 1 H), 8.77 (s, 1 H), 8.68 (d, J = 6.4 Hz, 1 H), 8.63 (s, 1 H), 8.07 (d, J = 7.2 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.43 (s, 1 H), 7.20 (s, 2 H), 6.49 (s, 1 H), 5.03-4.99 (m, 1 H), 3.40-3.31 (m, 2 H), 3.06-2.96 (m, 2 H), 1.27 (s, 9 H).

Example 69: Preparation of compound A29-022

SP-0010529-125 SP-0010529-128 SP-0010529-140 A29-022

To a suspension of potassium tert-butanolate (2.9 g, 26 mmol) in dry ethyl ether (100 mL) at 0 °C was added 5-fluoro-2,3-dihydroinden-l-one (3.0 g, 20 mmol) dropwise. The mixture was stirred at 0 °C for 0.5 h. Then the resulting mixture was added t-butyl nitrite (2.67 g, 26 mmol) slowly and the reaction mixture was stirred for additional 1 h. The mixture was poured into ice-cooled water (300 mL) and extracted with EtOAc (1000 mL). The organic layer was dried over anhydrous Na₂S0₄, filtered and concentrated. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 2: 1) to give the desired compound SP-0010529-118 (a mixture of two isomers), (2.0 g, yield: 57%) as a brown color solid. LC-MS 180 (M+H)⁺, purity 100% (UV 214 nm).

To a solution of compound SP-0010529-118 in MeOH / AcOH (30 mL / 2.0 mL) were added Pd/C (300 mg, 10%>) and (Boc)₂0 (4.9 g, 22.8 mmol). The mixture was stirred at room temperature overnight under H₂ atmosphere. The resulting mixture was filtered off and the filtrate was evaporated to give the crude product which was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 3: 1) to give the desired compound SP-0010529-120 (1.2 g, yield: 39%) as a pink color solid. LC-MS 210 (M- 56+H)⁺, purity 53% (UV 214 nm).

To a solution of compound SP-0010529-120 (1.2 g, 4.52 mmol) in THF (40 mL) were added DMAP (248 mg, 2.0 mmol) and (Boc)₂0 (436 mg, 2.0 mmol). The mixture was stirred at room temperature overnight. The reation mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (using petroleum ether:EtOAc = 20: 1 - 10: 1) to give the desired compound SP-0010529-125 (1.3 g, yield: 78%) as colorless oil. LC-MS 210 (M-56-100+H)⁺, purity 55% (UV 214 nm).

To a solution of compound SP-0010529-125 (1.3 g, 3.5 mmol) in MeOH (50 mL) was added Pd/C (300 mg, 10%>). The mixture was stirred at room temperature for overnight under H₂ atmosphere. The resulting mixture was filtered through celite and the filtrate was concentrated under reduced pressure to provide the crude compound SP-0010529-128 (1.5g of the crude) which was used directly for the next step of reaction without purification. LC- MS 196 (M-156+H)⁺, purity 44% (UV 214 nm).

A mixture of compound SP-0010529-128 (1.5 g of the crude) in HC1 / 1,4-dioxane (4 Μ,ΙΟ.Ο mL) was stirred at room temperature for 20 h. All of the volatiles were removed under reduced pressure to give the residue. The crude mixture was dissolved in water (20 mL), and EtOAc (20 mL) was added to extract the by-product. The aqueous phase was dried to provide the desired compound SP-0010529-140 (380 mg of the crude) as a form of HC1 salt. LC-MS 152(M+H)⁺, purity 65% (UV 214 nm).

To a solution of compound SP-0010529-140 (75 mg, 0.4 mmol) in i-PrOH (10 mL) were added 4-chloro-6-fluoroquinazoline (55 mg, 0.3 mmol) and TEA (0.5 mL). The mixture was stirred at 85 °C for 3 h. The resulting mixture was evaporated under reduced pressure and the residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 5: 1 - 1 : 1) to give the target compound A29-022 as a white solid

(20 mg, yield: 22%). LC-MS 298 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (DMSO- d₆, 400 MHz) δ 8.51 (s, 1 H), 8.28-8.20 (m, 2 H), 7.79-7.75 (dd, Ji = 5.6 Hz , J₂ = 9.2 Hz, 1 H), 7.69-7.67 (dd, Ji = 2.8 Hz, J₂ = 8.4 Hz, 1 H), 7.29-7.26 (dd, Ji = 2.4 Hz, J₂ = 8.4 Hz , 1 H), 7.12-7.09 (dd, Ji = 2.4 Hz, J₂ = 9.2 Hz, 1 H), 7.05-6.98 (m, 1 H), 5.04-5.00 (m, 1 H), 3.40-3.34 (m, 2 H), 3.08-3.01 (m, 2 H).

Example 70: Preparation of compound A29-023, A29-033 and A29-035

A29-035 A29-033 To a solution of 5-hydroxy-2,3-dihydroinden-l-one (1.48 g, 10.0 mmol) in dry DMF (20 mL) were added iodomethane (7.1 g, 50.0 mmol) and K₂CO₃ (2.76 g, 20 mmol). The mixture was stirred at room temperature for 20 h under N₂ atmosphere. The resulting mixture was quenched by water (100 mL), extracted with EtOAc (2 x 100 mL) and washed with brine (50 mL). The combined organic layer was dried over anhydrous Na₂S0₄, filtered and evaporated to give the crude compound SP-0010529-138 as a brown solid (1.9 g of the crude). LC-MS 163 (M+H)⁺, purity 95% (UV 214 nm).

To a suspension of compound SP-0010529-138 (1.9 g of the crude) in EtOH/H₂0 (10 mL / 4.0 mL) were added a solution of hydro xylamine hydrochloride (1.72 g, 25 mmol) in EtOH/H₂0 (10 mL / 4.0 mL) and sodium acetate (2.05 g, 25.0 mmol). The mixture was refluxed for 4.0 h under N₂ atmosphere. The reaction mixture was poured into water (100 mL). The suspension was filtered off and the cake was the crude compound SP-0010529- 139 as a brown solid (1.8 g of the crude). LC-MS 178 (M+H)⁺, purity 73% (UV 214 nm);

To a solution of compound SP-0010529-139 (354 mg, 2.0 mmol) in MeOH (20 mL) were added Raney-Nickel (200 mg) and NH₃.H₂0 (1.0 mL). The mixture was stirred at room temperature overnight under H₂ atmosphere. The resulting mixture was filtered through celite and the filtrate was evaporated to provide the crude compound 3 (340 mg of crude), which was used directly for the next step of reaction without purification. LC-MS 147 (M-NH₃+H)⁺, purity 95% (UV 214 nm).

To a solution of compound SP-0010529-147 (326 mg, 2.0 mmol) in i-PrOH (10 mL) were added 4-chloro-6-fluoroquinazoline (364 mg, 2.0 mmol) and TEA (1.5 mL). The mixture was stirred at 85 °C for 3.0 h. The resulting mixture was evaporated under reduced pressure and the residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 5: 1 - 2: 1) to give compound A29-023 as a white solid (200 mg, yield: 32%). LC-MS 310 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (DMSO-d₆, 400 MHz) δ 8.51 (s, 1 H), 8.37 (d, J = 8.0 Hz, 1 H), 8.24-8.21 (dd, Ji = 2.8 Hz, J₂ = 10.0 Hz, 1 H), 7.70-7.65 (m, 1 H), 8.16 (d, J = 8.4 Hz, 1 H), 6.88 (s, 1H), 6.75-6.72 (dd, Ji = 2.4 Hz, J₂ = 8.4 Hz, 1 H), 5.93-5.90 (m, 1 H), 3.73 (s, 3 H), 3.03-2.99 (m, 2 H), 2.88-2.84 (m, 1 H), 2.57-2.53 (m, 1 H), 2.08-2.03 (m, 1 H).

To a solution of compound A29-023 (80 mg, 0.26 mmol) in DCM (10 mL) was added BBr₃ (323 mg, 1.29 mmol) slowly at -10 °C. The mixture was stirred at 0 °C for 2.0 h. The eaction was quenched with a saturated aqueous NaHC0₃ (10 mL) and extracted with DCM (50 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give the title compound A29- 046 as a white solid (50 mg, yield: 65%). LC-MS 296 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (DMSO-de, 400 MHz) δ 9.27 (s, 1 H), 8.50 (s, 1 H), 8.34 (d, J = 8.0 Hz, 1 H), 8.24-8.21 (dd, Ji = 2.8 Hz, J₂ = 10.0 Hz, 1 H), 7.78-7.75 (dd, Ji = 6.4 Hz, J₂ = 9.2 Hz, 1 H), 7.69-7.64 (m, 1 H), 7.04 (d, J = 8.0 Hz, 1 H), 6.68 (s, 1 H), 6.58-6.56 (dd, Ji = 2.4 Hz, J₂ = 8.0 Hz, 1 H), 5.87 (d, J = 7.6 Hz, 1 H), 2.96-2.93 (m, 1 H), 2.82-2.78 (m, 1 H), 2.54-2.48 (m, 1 H), 2.04-1.99 (m, 1 H).

To a solution of compound A29-046 (14 mg, 0.05 mmol) and 4-(2-chloroethyl) morpholine hydrochloride (32 mg, 0.15 mmol) in DMF (5 mL) were added TBAI (3 mg, 0.01 mmol) and K₂C0₃ (41 mg, 0.3 mmol). The mixture was stirred at 90 °C for 3.0 h. Water (20 mL) was added to dilute the resulting mixture and the suspension was extracted with EtOAc (2 x 50 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 5: 1 - 2:1) to give the target compound A29-047 as a white solid (6.0 mg, yield: 29%). LC-MS 409 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (DMSO-d₆, 400 MHz) δ 8.49 (s, 1 H), 8.00 (d, J = 6.4 Hz, 1 H), 7.80-7.76 (m, 1 H), 7.64-7.59 (m, 1 H), 7.21 (d, J = 5.6 Hz, 2 H), 6.89 (s, 1 H), 6.79 (d, J = 8.0 Hz, 1 H), 5.98 (t, J = 7.2 Hz, 1 H), 3.14 (t, J = 5.6 Hz, 2 H), 3.73 (t, J = 4.8 Hz, 4 H), 3.10-3.07 (m, 1 H), 2.95-2.91 (m, 1 H), 2.81 (t, J = 5.6 Hz, 2 H), 2.70-2.66 (m, 2 H), 2.65-2.60 (m, 4 H), 2.14-2.09 (m, 1 H).

Example 71: Preparation of compound A29-024

To a solution of A23-001 (130 mg, 0.43 mmol) in DMF (2.0 mL) were added furan- 2-carboxylic acid (73 mg, 0.65 mmoL), HATU (490 mg, 1.29 mmol) and triethylamine (0.31 mL, 2.15 mmol). The reaction mixture was stirred at 50 °C for 8 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-024 as a white solid (56 mg, yield: 33%). LC-MS 396 (M+H) ⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 10.11 (s, 1H), 8.99 (s, 1 H), 8.69 (d, J = 6.4 Hz, 1 H), 8.63 (s, 1 H), 8.07 (d, J = 6.8 Hz, 1 H), 7.93 (s, 1 H), 7.79 (d, J = 8.4 Hz, 1 H), 7.72 (s, 1 H), 7.52 (d, J = 6.0 Hz, 1 H), 7.32 (d, J = 3.6 Hz, 1 H), 7.23 (d, J = 8.4 Hz, 1 H), 6.70-6.69 (m, 3 H), ), 5.04-5.00 (m, 1 H), 4.68 (s, 2 H), 3.41-3.39 (m, 2 H), 3.06-2.97 (m, 2 H).

Example 72: Preparation of compound A29-025

A02-171 A29-025

To a solution of compound A02-171 (100 mg, 0.340 mmol) and triethylamine (343 mg, 3.40 mmol) in DCM (6 mL) cooled in an ice-bath was added isobutyryl chloride (43 mg, 0.408 mmol). The reaction mixture was stirred at room temperature for 2 h. Water (0.1 mL) was added to quench the reaction. The mixture was stirred for 10 min and concentrated under reduced presure. The residue was purified by Pre-HPLC to give A29-025 (30 mg, yield: 24%). LC-MS 365 (M+H)⁺, purity 99% (UV 214 nm); 1H NMR (400 MHz, DMSO- d6) δ 9.74 (s, 1 H), 8.51 (s, 1 H), 8.27-8.22 (m, 2 H), 7.78-7.68 (m, 2 H), 7.59 (s, 1 H), 7.35 (d, J = 7.6 Hz, 1 H), 7.16 (d, J = 8.0 Hz, 1 H), 5.01-4.90 (m, 1 H), 3.00 (td, J = 16.4 Hz, 5.6 Hz, 2 H), 2.59-2.50 (m, 3 H), 1.10-1.08 (m, 6 H).

Example 73: Preparation of compound A29-026

A02-171 A29-026

To a solution of compound A02-171 (100 mg, 0.340 mmol) in DMF (3 mL) were added 1-hydroxycyclopropanecarboxylic acid (52 mg, 0.510 mmol), HATU (190 mg, 0.510 mmol) and triethylamine (343 mg, 3.40 mmol). The mixture was stirred at 40 °C for 16 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (2 x 20 mL). The organic layer was washed with water (10 mL) and brine (10 mL), dried over Na₂S0₄, filtered, and concentrated. The residue was purified by Prep-HPLC to give A29- 026 as a white solid (50 mg, yield: 39%). LC-MS 379 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.67 (s, 1 H), 8.51 (s, 1 H), 8.28 (d, J = 6.8 Hz, 1 H), 8.23 (dd, J = 10.0 Hz, 2.4 Hz, 1 H), 7.77 (dd, J = 9.2 Hz, 5.6 Hz, 1 H), 7.70-7.66 (m, 2 H), 7.48 (dd, J = 8.4 Hz, 2.0 Hz, 1 H), 7.17 (d, J = 8.4 Hz, 1 H), 6.55 (s, 1 H), 5.03-4.98 (m, 1 H), 3.38-3.30 (m, 2 H), 3.05-2.96 (m, 2 H), 1.15-1.13 (m, 2 H), 0.96-1.93 (m, 2 H).

Example 74: Preparation of compound A29-027

A02-171 A29-027

To a solution of compound A02-171 (100 mg, 0.340 mmol) in DCM (6 mL) cooled in an ice-bath were added methanesulfonyl chloride (58 mg, 0.510 mmol) and triethylamine (343 mg, 3.40 mmol). The reaction mixture was stirred at room temperature for 3 h. Water (0.1 mL) was added to quench the reaction. The mixture was stirred for additional 10 min and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give A29-027 (15 mg, yield: 14%). LC-MS 373 (M+H)⁺, purity 99% (UV 214 nm); 1H NMR (400 MHz, MeOD-d4) δ 8.50 (s, 1 H), 8.03 (dd, J = 9.6 Hz, 2.8 Hz, 1 H), 7.78 (dd, J = 9.2 Hz, 5.2 Hz, 1 H), 7.62 (td, J = 8.0 Hz, 2.8 Hz, 1 H), 7.24-7.21 (m, 2 H), 7.07 (dd, J = 8.4 Hz, 2.4 Hz, 1 H), 5.16-5.12 (m, 1 H), 3.44-3.41 (m, 2 H), 3.12-3.03 (m, 2 H), 2.94 (s, 3 H). Example 75: Preparation of compound A29-028 and A29-030

A29-028 A29-030

To a solution of 5-fluoro-2,3-dihydro-lH-inden-2-amine hydrochloride (75 mg, 0.4 mmol) in i-PrOH (10 mL) were added 4-chloro-6-iodoquinazoline (116 mg, 0.4 mmol) and TEA (0.5 mL). The mixture was stirred at 85 °C for 3 h. All of the volatiles were evaporated and the residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 3:1) to give the desired compound A29-028 as a white solid (50 mg, yield: 30%). LC-MS 406 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (DMSO-de, 400 MHz) δ 8.78 (d, J = 6.0, 1 H), 8.53 (s, 1 H), 8.45 (d, J = 6.8 Hz, 1 H), 8.03- 8.00 (m, Ji = 2.0 Hz, J₂ = 8.8 Hz, 1 H), 7.47 (d, J = 8.8 Hz, 1 H), 7.29-7.26 (m, 1 H), 7.12- 7.09 (dd, Ji = 2.4 Hz, J₂ = 9.2 Hz, 1 H), 6.99 (t, J = 9.6 Hz, 1 H), 5.03-5.00 (m, 1 H), 3.41- 3.33 (m, 2 H), 3.09-3.02 (m, 2 H). To a solution of compound A29-028 (121 mg, 0.30 mmol) in dry DMA (4.0 mL) was added CuCN (138 mg, 1.5 mmol). The mixture was stirred at 110 °C for 20 h under N₂ atmosphere. The resulting mixture was quenched with water (20 mL), extracted with EtOAc (3 x 30 mL) and washed with brine (20 mL). The combined organic layer was dried over anhydrous Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 2: 1) to give the title compound A29-030 as a white solid (40 mg, yield: 43%). LC-MS 305 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (DMSO-d₆, 400 MHz) δ 8.97 (s, 1 H), 8.68- 8.66 (m, 1 H), 8.62 (s, 1 H), 8.08-8.05 (m, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.30-7.27 (m, 1 H), 7.13-7.10 (m, 1 H), 7.02-6.97 (m, 1 H), 5.06-5.00 (m, 1 H), 3.44-3.33 (m, 2 H), 3.08- 2.98 (m, 2 H).

Example 76: Preparation of compound A22-148

A29-028 A22-148

To a solution of N-(5-fluoro-2,3-dihydro-lH-inden-2-yl)-6-iodoquinazolin-4-amine (80 mg, 0.2 mmol) in dry DMSO (4.0 mL) were added Cul (19 mg, 0.1 mmol) and sodium methanesulfmate (118 mg, 1.0 mmol). The mixture was stirred at 110 °C for 20 h under N₂ atmosphere. The resulting mixture was quenched with water (20 mL), extracted with EtOAc (2 x 50 mL) and washed with brine (20 mL). The combined organic layer was dried over anhydrous Na₂S0_4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 1 : 1 as eluent) to give the target compound A22-148 (25 mg, yield: 34%) as a white solid. LC- MS 358 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (DMSO- ₆, 400 MHz) δ 9.03 (s, 1 H), 8.95 (d, J = 6.4 Hz, 1 H), 8.65 (s, 1 H), 8.19 (d, J = 7.6 Hz, 1 H), 7.88 (d, J = 7.6 Hz, 1 H), 7.30-7.27 (m, 1 H), 7.11 (d, J = 7.6 Hz, 1 H), 7.03-6.99 (m, 1 H), 5.10-5.07 (m, 1 H), 3.44-3.35 (m, 2 H), 3.27 (s, 3 H), 3.13-3.02 (m, 2 H). Example 77: Preparation of compound A29-029, A29-031, A29-055, A29-092

A29-055 A29-092

To a solution of 5-methoxy-2,3-dihydro-lH-inden-l -amine (526 mg, 2.0 mmol) in i- PrOH (20 mL) were added 4-chloro-6-iodoquinazoline (580 mg, 2.0 mmol) and TEA (1.0 mL). The mixture was stirred at 85 °C for 3 h. All of the volatiles were evaporated and the residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 2: 1) to give the desired compound A29-029 as a pale yellow solid (400 mg, yield: 48%). LC-MS 418 (M+H)⁺, purity 97% (UV 214 nm); 1H NMR (DMSO-d₆, 400 MHz) δ 8.80 (s, 1 H), 8.55-8.52 (m, 2 H), 8.03-8.00 (dd, Ji = 2.0 Hz, J₂ = 8.4 Hz, 1 H), 7.47 (d, J = 8.8 Hz, 1H), 7.16 (d, J = 8.4 Hz, 1 H), 6.88 (s, 1 H), 6.75-6.72 (dd, Ji = 2.4 Hz, J₂ = 8.4 Hz, 1 H), 5.93-5.90 (m, 1 H), 3.73 (s, 3 H), 3.03-3.00 (m, 1 H), 2.88-2.83 (m, 1 H), 2.56-2.51 (m, 1 H), 2.07-2.03 (m, 1 H).

To a solution of compound A29-029 (83 mg, 0.2 mmol) in dry DMSO (3.0 mL) were added Cul (4 mg, 0.02 mmol) and sodium methanesulfmate (118 mg, 1.0 mmol). The mixture was stirred at 110 °C for 20 h under N₂ atmosphere. The resulting mixture was quenched with water (20 mL), extracted with EtOAc (2 x 50 mL) and washed with brine (20 mL). The combined organic layer was dried over anhydrous Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 1 : 1) to give the title compound A29-031 (42 mg, yield: 56%) as a white solid. LC-MS 370 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (DMSO- ₆, 400 MHz) δ 9.06-9.04 (m, 2 H), 8.63 (s, 1 H), 8.20-8.17 (m, 1 H), 7.87 (d, J = 7.2 Hz, 1 H), 7.18 (d, J = 8.4 Hz, 1 H), 6.90 (d, J = 2.0 Hz, 1 H), 6.76-6.73 (m, 1 H), 5.99-5.96 (m, 1 H), 3.74 (s, 3 H), 3.26 (s, 3 H), 3.04-3.01 (m, 1 H), 2.90-2.85 (m, 1 H), 2.57-2.50 (m, 1 H), 2.12-2.07 (m, 1 H).

To a solution of compound A29-031 (150 mg, 0.4 mmol) in DCM (20 mL) was added BBr₃ (500 mg, 2.0 mmol) slowly at -10 °C. The mixture was stirred at 0 °C for 2.0 h. The raction was quenched with a saturated aquesous NaHC0₃ (20 mL) and extracted with DCM (100 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give the compound, A29-055 as a white solid (35 mg, yield: 24%). LC-MS 356 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (DMSO-d₆, 400 MHz) δ 9.29 (s, 1 H), 9.04 (s, 1 H), 9.04-9.01 (m, 1 H), 8.61 (s, 1 H), 8.19-8.16 (dd, Ji = 2.4 Hz, J₂ = 8.8 Hz, 1 H), 7.86 (d, J = 8.8 Hz, 1 H), 7.06 (d, J = 8.4 Hz, 1 H), 6.69 (s, 1 H), 6.59-6.57 (dd, Ji = 2.0 Hz, J₂ = 8.0 Hz, 1 H), 5.95-5.92 (m, 1 H), 3.26 (s, 3 H), 2.98-2.95 (m, 1 H), 2.83-2.79 (m, 1 H), 2.54-2.50 (m, 1 H), 2.08-2.03 (m, 1 H).

To a solution of compound A29-055 (25 mg, 0.07 mmol) and 4-(2-chloroethyl) morpholine hydrochloride (21 mg, 0.1 mmol) in DMF (6 mL) were added TBAI (9 mg, 0.02 mmol) and K₂C0₃ (34 mg, 0.25 mmol). The mixture was stirred at 90 °C for 3.0 h. Water (30 mL) was added to dilute the reaction mixture. The suspension was extracted with EtOAc (2 x 50 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 5: 1 - 2: 1) to give the title A29-092 as a white solid (15 mg, yield: 58%). LC-MS 469 (M+H)⁺, purity 97% (UV 214 nm). 1H NMR (DMSO-d₆, 400 MHz) δ 8.79 (s, 1 H), 8.35 (s, 1 H), 8.15-8.13 (dd, Ji = 2.0 Hz, J₂ = 8.8 Hz, 1 H), 7.98 (d, J = 8.8 Hz, 1 H), 7.27-7.25 (m, 1 H), 6.87 (d, J = 2.0 Hz, 1 H), 6.80-6.77 (dd, Ji = 2.4 Hz, J₂ = 8.8 Hz, 1 H), 6.29-6.26 (m, 1 H), 5.94-5.91 (m, 1 H), 4.16-4.13 (m, 2 H), 3.78-3.73 (m, 4 H), 3.10-3.05 (m, 1 H), 3.08 (s, 3 H), 2.98-2.95 (m, 1 H), 2.86-2.83 (m, 2 H), 2.76-2.74 (m, 1 H), 2.62 (m, [2+2] H), 2.11-2.08 (m, 1 H).

Example 78: Preparation of compound A29-036

A02-171 A29-036

To a solution of compound A02-171 (100 mg, 0.340 mmol) in DMF (3 mL) wer added 2-phenoxyacetic acid (62 mg, 0.408 mmol), HATU (168 mg, 0.442 mmol) and triethylamine (343 mg, 3.40 mmol). The mixture was stirred at 40 °C for 5 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (2 x 20 mL). The organic layer was washed with water (10 mL) and brine (10 mL). The combined organic layers were dried over Na₂S0₄, filtered, and concentrated. The residue was purified by Prep- HPLC to give A29-036 as a white solid (45 mg, yield: 31%). LC-MS 429 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 10.01 (s, 1 H), 8.51 (s, 1 H), 8.27 (d, J = 6.8 Hz, 1 H), 8.22 (dd, J = 10.4 Hz, 2.8 Hz, 1 H), 7.77 (dd, J = 9.2 Hz, 5.6 Hz, 1 H), 7.67 (td, J = 8.8 Hz, 2.8 Hz, 1 H), 7.61 (s, 1 H), 7.40 (dd, J = 8.4 Hz, 1.6 Hz, 1 H), 7.35- 7.30 (m, 2 H), 7.21 (d, J = 8.0 Hz, 1 H), 7.01-6.96 (m, 3 H), 5.03-4.98 (m, 1 H), 4.68 (s, 2 H), 3.38-3.30 (m, 2 H), 3.05-2.96 (m, 2 H).

Example 79: Preparation of compound A29-037

A02-171 A29-037

To a solution of compound A02-171 (100 mg, 0.340 mmol) in DMF (3 mL) were added furan-2-carboxylic acid (46 mg, 0.408 mmol), HATU (168 mg, 0.442 mmol) and triethylamine (343 mg, 3.40 mmol). The mixture was stirred at 40 °C for 5 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAC (2 x 20 mL). The organic layer was washed with water (10 mL) and brine (10 mL). The combined organic layers were dried over Na₂S0₄, filtered, and concentrated. The residue was purified by Prep-HPLC to give A29-037 as a white solid (35 mg, yield: 27%). LC-MS 389 (M+H)⁺, purity 97% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 10.11 (s, 1 H), 8.52 (s, 1 H), 8.29 (d, J = 6.8 Hz, 1 H), 8.24 (dd, J = 10.0 Hz, 2.8 Hz, 1 H), 7.93 (s, 1 H), 7.77 (dd, J = 5.6 Hz, 2.8 Hz, 1 H), 7.74-7.65 (m, 2 H), 7.52 (dd, J = 8.4 Hz, 2.0 Hz, 1 H), 7.32 (d, J = 3.2 Hz, 1 H), 7.22 (d, J = 8.0 Hz, 1 H), 6.70 (dd, J = 3.6 Hz, 2.0 Hz, 1 H), 5.03-4.98 (m, 1 H), 3.41- 3.36 (m, 2 H), 3.03 (td, J = 16.8 Hz, 6.0 Hz, 2 H).

Example 80: Preparation of compound A29-038

A02-171 A29-038

To a solution of compound A02-171 (100 mg, 0.340 mmol) and pyridine (40 mg, 0.510 mmol) in DMF (3 mL) was phenyl 5-tert-butylisoxazol-3-ylcarbamate (96 mg, 0.370 mmol). The reaction mixture was stirred at 50 °C for 16 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (2 x 20 mL). The organic layer was washed with HC1 (1M, 10 mL) and brine (10 mL). The combined organic layers were dried over Na₂S0₄, filtered, and concentrated under reduced pressure. The residue was purified by Prep-HPLC to give A29-038 as a white solid (30 mg, yield: 19%). LC-MS 461 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, MeOD-d4) δ 8.50 (s, 1 H), 8.04 (dd, J = 9.6 Hz, 2.4 Hz, 1 H), 7.78 (dd, J = 9.2 Hz, 5.2 Hz, 1 H), 7.62 (td, J = 8.0 Hz, 2.8 Hz, 1 H), 7.44 (s, 1 H), 7.22 (s, 2 H), 6.38 (s, 1 H), 5.18-5.10 (m, 1 H), 3.49-3.41 (m, 2 H), 3.08 (td, J = 15.6 Hz, 6.4 Hz, 2 H), 1.35 (s, 9 H).

Example 81: Preparation of compound A29-039

A29-039

To a solution of compound A02-171 (150 mg, 0.510 mmol) and triethylamine (515 mg, 5.10 mmol) in DCM (6 mL) cooled at 0 °C was added 2-chloroacetyl chloride (75 mg, 0.663 mmol). The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was partitioned between aqueous phase (20 mL) and DCM phase (20 mL). The organic layer was washed with aqueous HC1 (1M, 10 mL) and brine (10 mL). The combined organic layers were dried over Na₂S0₄, filtered, and concentrated to give the crude product SP-0010485-180 -1 (140 mg), which was used in the next step of reaction without further purification.

To a solution of SP-0010485-180 -1 (140 mg, 0.510 mmol, crude) in DMF (3.0 mL) were added KI (8 mg, 0.05 mmol) and imidazole (138 mg, 2.04 mmol). The mixture was stirred at 120 °C for 2 h under microwave. The resulting mixture was diluted by water (20 mL), extracted with EtOAc (2 x 30 mL) and washed with water (20 mL) and brine (20 mL). The combined organic layer was dried over anhydrous Na₂S0₄. The residue was purified by Prep-HPLC to give the compound A29-039 (45 mg, yield: 22%, two steps) as a white solid. LC-MS 403 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6): 510.24 (s, 1 H), 8.52 (s, 1 H), 8.27 (d, J = 6.8 Hz, 1 H), 8.23 (dd, J = 10.4 Hz, 2.8 Hz, 1 H), 7.77 (dd, J = 9.2 Hz, 5.2 Hz, 1 H), 7.68 (td, J = 8.4 Hz, 2.8 Hz, 1 H), 7.63 (s, 1 H), 7.54 (s, 1 H), 7.35 (dd, J = 8.4 Hz, 2.0 Hz, 1 H), 7.21 (d, J = 8.0 Hz, 1 H), 7.16 (s, 1 H), 6.90 (s, 1 H), 5.03-4.98 (m, 1 H), 4.89 (s, 2 H), 3.39-3.31 (m, 2 H), 3.05-2.97 (m, 2 H).

Example 82: Preparation of compound A29-040

A02-171 A29-040

To a solution of compound A02-171 (100 mg, 0.340 mmol) in DMF (3 mL) were added 2-morpholinoacetic acid (74 mg, 0.510 mmol), HATU (193 mg, 0.510 mmol) and triethylamine (343 mg, 3.40 mmol). The mixture was stirred at 40 °C for 5 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (2 x 20 mL). The organic layer washed with water (10 mL) and brine (10 mL). The combined organic layers were dried over Na₂S0₄, filtered, and concentrated under reduced pressure. The residue was purified by Prep-HPLC to give A29-040 as a white solid (40 mg, yield: 28%). LC-MS 422 (M+H)⁺, purity 97% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.50 (s, 1 H), 8.03 (dd, J = 10.0 Hz, 3.2 Hz, 1 H), 7.78(dd, J = 8.8 Hz, 5.2 Hz, 1 H), 7.62 (td, J = 8.4 Hz, 4.2 Hz, 1 H), 7.55 (s, 1 H), 7.36 (dd, J = 8.0 Hz, 2.0 Hz, 1 H), 7.23 (d, J = 8.0 Hz, 1 H), 5.16- 5.12 (m, 1 H), 3.80-3.78 (m, 4 H), 3.49-3.41 (m, 2 H) , 3.19 (s, 2 H), 3.13-3.04 (m, 2 H), 2.63-2.61 (m, 4 H).

Example 83: Preparation of compound A29-041

A02-171 A29-041

To a solution of compound A02-171 (100 mg, 0.340 mmol) in DMF (3 mL) were added cyclopropanecarboxylic acid (44 mg, 0.510 mmol), HATU (193 mg, 0.510 mmol) and triethylamine (343 mg, 3.40 mmol). The mixture was stirred at 40 °C for 5 h. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (2 x 30 mL). The organic layer washed with water (20 mL) and brine (20 mL). The combined organic layers were dried over Na₂S0₄, filtered, and concentrated. The residue was purified by

Prep-HPLC to give A29-041 as a white solid (45 mg, yield: 37%). LC-MS 363 (M+H)⁺, purity 98% (UV 214 nm); 1H NMR (400 MHz, MeOD-d4) δ 8.50 (s, 1 H), 8.03 (dd, J = 10.0 Hz, 3.2 Hz, 1 H), 7.78(dd, J = 8.8 Hz, 5.2 Hz, 1 H), 7.62 (td, J = 9.2 Hz, 3.2 Hz, 1 H), 7.52 (s, 1 H), 7.31 (dd, J = 8.0 Hz, 2.0 Hz, 1 H), 7.20 (d, J = 8.0 Hz, 1 H), 5.16-5.12 (m, 1 H), 3.47-3.40 (m, 2H), 3.11-3.02 (m, 2 H), 1.79-1.75 (m, 1 H), 0.98-0.94 (m, 2 H), 0.88- 0.84 (m, 2 H).

Example 84: Preparation of compound A29-053

To a solution of compound A29-003 (100 mg, 0.231 mmol) and zinc powder (300 mg, 4.62 mmol) in THF (6 mL) was added a solution of ammonium chloride (184 mg, 3.47 mmol) in water (3 mL). The reaction mixture was stirred at 60 °C for 2 h. The reaction mixture was partitioned between water (10 mL) and EtOAc (30 mL). The organic layer was washed with brine (20 mL). The combined organic layer was dried over Na₂S0₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 1 : 1) to afford A29-053 (48 mg, yield:

75%). LC-MS 277 (M+H)⁺, purity 98% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.50 (s, 1 H), 8.35 (d, J = 8.0 Hz, 1 H), 8.27 (d, J = 6.8 Hz, 1 H), 7.79-7.74 (m, 1 H), 7.68 (dd, J = 8.4 Hz, 1.2 Hz, 1 H), 7.49 (td, J = 6.8 Hz, 1.2 Hz, 1 H), 6.88 (d, J = 8.0 Hz, 1 H), 6.47 (s, 1 H), 6.39 (dd, J = 8.0 Hz, 2.0 Hz, 1 H), 5.00-4.94 (m, 1 H), 4.85 (s, 2 H), 3.25-3.17 (m, 2 H), 2.90 (td, J = 16.4 Hz, 6.8 Hz, 2 H). Example 85: Preparation of compound A29-054

To a solution of compound A29-053 (100 mg, 0.362 mmol) in DMF (3 mL) were added 2-hydroxyacetic acid (41 mg, 0.543 mmol), HATU (206 mg, 0.543 mmol) and triethylamine (365 mg, 3.62 mmol). The mixture was stirred at 40 °C for 5 h. The reaction mixture was poured into water (15 mL) and extracted with EtOAc (2 x 20 mL). The organic layer was washed with water (15 mL) and brine (15 mL). The combined organic layer was dried over Na₂S0₄, filtered, and concentrated under reduced pressure. The residue was purified by Prep-HPLC to give A29-054 as a white solid (40 mg, yield: 33%). LC-MS 335 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.56 (s, 1 H), 8.51 (s, 1 H), 8.35-8.32 (m, 2 H), 7.76 (td, J =8.4 Hz, 1.2 Hz, 1 H), 7.69 (dd, J = 8.0 Hz, 1.2 Hz, 1 H), 7.65 (s, 1 H), 7.49 (td, J = 8.0 Hz, 1.2 Hz, 1 H), 7.44 (dd, J = 8.0 Hz, 1.2 Hz, 1 H), 7.17 (d, J = 8.0 Hz, 1 H), 5.65 (t, J = 6.0 Hz, 1 H), 5.06-5.00 (m, 1 H), 3.98 (d, J = 6.0 Hz, 2 H), 3.37-3.30 (m, 2 H), 3.02 (td, J = 16.4 Hz, 6.8 Hz, 2 H).

Example 86: Preparation of compound A29-060

A02-171 A29-060

To a solution of compound A02-171 (100 mg, 0.272 mmol) in DCM (6 mL) cooled in an ice-bath were added trifluoroacetic anhydride (86 mg, 0.408 mmol), triethylamine (410 mg, 4.08 mmol) and DMAP (3 mg, 0.027 mmol). After stirring at room temperature for 2 h, the reaction mixture was concentrated under reduced pressure. The residue was purified by Pre-HPLC to give A29-060 (80 mg, yield: 60%). LC-MS 391 (M+H)⁺, purity 99% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 11.20 (s, 1 H), 8.57 (s, 1 H), 8.49 (d, J = 6.4 Hz, 1 H), 8.26 (dd, J = 10.0 Hz, 2.8 Hz, 1 H), 7.78 (dd, J = 9.2 Hz, 5.6 Hz, 1 H), 7.72 (td, J = 8.8 Hz, 2.8 Hz, 1 H), 7.60 (s, 1 H), 7.44 (dd, J = 8.4 Hz, 2.0 Hz, 1 H), 7.29 (d, J = 8.0 Hz, 1 H), 5.06-5.03 (m, 1 H), 3.43-3.36 (m, 2 H), 3.11-3.02 (m, 2 H). Example 87: Preparation of compound A29-061

A02-171 A29-061

To a solution of compound A02-171 (150 mg, 0.510 mol) in DMF (3 mL) were added 3-hydroxy-3-methylbutanoic acid (120 mg, 1.020 mmol), HATU (388 mg, 1.020 mmol) and triethylamine (515 mg, 5.10 mmol). The mixture was stirred at 40 °C for 16 h. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (2 x 20 mL). The organic layer was washed with water (15 mL) and brine (15 mL). The combined organic layer was dried over Na₂S0₄, filtered, and concentrated. The residue was purified by Prep-HPLC to give A29-061 as a white solid (40 mg, yield: 20%). LC-MS 395 (M+H)⁺, purity 97% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.77 (s, 1 H), 8.52 (s, 1 H), 8.26 (d, J = 6.8 Hz, 1 H), 8.23 (dd, J = 10.0 Hz, 2.8 Hz, 1 H), 7.77 (dd, J = 9.2 Hz, 5.6 Hz, 1 H), 7.67 (td, J = 8.4 Hz, 2.8 Hz, 1 H), 7.58 (s, 1 H), 7.32 (dd, J = 8.4 Hz, 2.0 Hz, 1 H), 7.17 (d, J = 8.0 Hz, 1 H), 5.03-4.97 (m, 1 H), 4.76 (s, 1 H), 3.37-3.30 (m, 2 H), 3.05-2.95 (m, 2 H), 2.41 (s, 2 H), 1.22 (s, 6 H).

Example 88: Preparation of compound A29-065

SP-0011321 -025 A29-065

A mixture of 2-amino-4-fluorobenzoic acid (1.55g, 10 mmol) in formamide (9.0 g, 0.2 mol) was heated at 200 °C for 20 h. After the reaction was completed, the mixture was cooled down, and filtered. The filtrate was washed with water (5 x 15 mL), dried over Na₂S0₄, filtered and concentrated under reduced pressure to afford crude product SP- 0011321-021 (1.07 g, yield: 65%). The resulting crude product was used in the next step of reaction without further purification.

To the crude product SP-0011321-021 (0.82 g, 5 mmol) was added thionyl dichloride (12.0 g, 0.1 mol) and catalytic amount of anhydrous DMF (0.5 mL). The reaction mixture was heated to reflux for 5 h. After the reaction was completed, the mixture was cooled down and excess thionyl dichloride was removed by rotary evaporation. The resulting crude product SP-0011321-023 (950 mg) was used in the next step of reaction without further purification.

To a solution of compound SP-0011321-023 (500 mg, 2.75 mmol) and 5-nitro-2,3- dihydro-lH-inden-2-amine (490 mg, 2.75 mmol) in isopropyl alcohol (30 mL) was added triethylamine (2.8 g, 27.5 mmol). The resulting solution was heated to 65 °C under N₂ for 2 h. The mixture was cooled down and excess of isopropyl alcohol was removed by rotary evaporation to give the crude compound SP-0011321-025 (898 mg).

To a solution of compound SP-0011321-025 (850 mg, 2.75 mmol) in MeOH (30 mL) was added 10% Pd/C (100 mg). The mixture was stirred at room temperature overnight under H₂ atmosphere. The resulting mixture was filtered through Celite and the filtrate was evaporated to give a residue, which was purified by silica gel column chromatography (using petroleum ether : EtOAc = 3: 1-2:2) to afford A29-065 (760 mg, yield: 94%). LC-MS 295 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- d6) δ 8.50-8.44 (m, 2 H), 8.37 (d, J = 6.8 Hz, 1 H), 7.43-7.38 (m, 2 H), 6.88 (d, J = 8.0 Hz, 1 H), 6.47 (s, 1 H), 6.39 (dd, J = 8.0 Hz, 2.0 Hz, 1 H), 5.00-4.94 (m, 1 H), 4.85 (s, 2 H), 3.25-3.17 (m, 2 H), 2.90 (td, J = 16.0 Hz, 6.8 Hz, 2 H).

Example 89: Preparation of compound A29-066

To a solution of compound SP-0011321-028 (120 mg, 0.408 mmol) in DCM (10 mL) cooled in an ice-bath were added acetic anhydride (83 mg, 0.816 mmol), triethylamine (410 mg, 4.08 mmol) and DMAP (4 mg, 0.041 mmol). The reaction mixture was stirred at room temperature for 2 h, and concentrated. The residue was purified by Pre-HPLC to give A29-066 (66 mg, yield: 44%). LC-MS 337 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.85 (s, 1 H), 8.52 (s, 1 H), 8.47-8.40 (m, 2 H), 7.55 (s, 1 H), 7.44- 7.39 (m, 2 H), 7.31 (dd, J = 8.4 Hz, 1.2 Hz, 1 H), 7.14 (d, J = 8.0 Hz, 1 H), 5.03-4.98 (m, 1 H), 3.36-3.28 (m, 2 H), 3.00 (td, J = 16.8 Hz, 6.4 Hz, 2 H), 2.03 (s, 3 H).

Example 90: Preparation of compound A29-032 and A29-050

^H | -O CuCN, DMA ^{H '} o⁷ BBr₃, DCM

A29-029 A29-032 A29-050

To a solution of A29-029 (125 mg, 0.3 mmol) in dry DMA (3 mL) was added CuCN (138 mg, 1.5 mmol). The mixture was stirred at 1 10 °C for 20 h under N₂ atmosphere. The resulting mixture was quenched by water (20 mL), extracted with EtOAc (3 x 30 mL) and washed with brine (20 mL). The combined organic layer was dried over anhydrous Na₂S0₄, filtered and concentrated. The residue was purified by silica gel column chromatography (using petroleum ether: EtOAc = 10: 1 - 2: 1) to give the title compound A29-032 as a white solid (48 mg, yield: 50%). LC-MS 317 (M+H)⁺, purity 99% (UV 214 nm); 1H NMR (DMSO-d₆, 400 MHz) δ 8.98 (d, J = 1.6 Hz, 1 H), 8.76 (d, J = 8.0 Hz, 1 H), 8.07-8.05 (dd, Ji = 1.6 Hz, J₂ = 8.8 Hz, 1 H), 7.80 (d, J = 8.4 Hz, 1 H), 7.19 (d, J = 8.4 Hz, 1 H), 6.90 (s, 1 H), 6.76-6.73 (m, 1 H), 5.92-5.90 (m, 1 H), 3.74 (s, 3 H), 3.04-3.01 (m, 1 H), 2.89-2.85 (m, 1 H), 2.59-2.53 (m, 1 H), 2.09-2.06 (m, 1 H).

To a solution of compound A29-032 (126 mg, 0.4 mmol) in DCM (10 mL) at -10 °C was added BBr₃ (500 mg, 2.0 mmol) slowly. The mixture was stirred at 0 °C for 1 h. The reaction was quenched with a saturated aqueous NaHC0₃ (20 mL) and extracted with DCM (100 mL). The DCM phase was evaporated and the residue was purified by Pre-HPLC to give the title compound A29-050 as a white solid (40 mg, yield: 33%). LC-MS 303 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (DMSO-d₆, 400 MHz) δ 9.30 (s, 1 H), 8.98 (s, 1 H), 8.72 (d, J = 7.6 Hz, 1 H), 8.61 (s, 1 H), 8.07-8.05 (dd, Ji = 1.6 Hz, J₂ = 8.8 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.07 (d, J = 8.4 Hz, 1 H), 6.69 (s, 1 H), 6.59-6.56 (m, 1 H), 5.89-5.86 (m, 1 H), 2.98-2.95 (m, 1 H), 2.84-2.79 (m, 1 H), 2.55-2.50 (m, 1 H), 2.05-2.02 (m, 1 H). Example 91: Preparation of compound A29-051

A29-029 A29-051

To a solution of compound A29-029 (41 mg, 0.1 mmol) in DCM (10 mL) at -10 °C was added BBr₃ (125 mg, 0.5 mmol) slowly. The mixture was stirred at 0 °C for 1 h. The reaction was quenched with a saturated solution of NaHC0₃ (10 mL) and extracted with DCM (50 mL). The organic phase was dried over Na₂S0₄, filtered, concentrated under reduced pressure. The residue was purified by Pre-HPLC to give the title compound A29- 051 as a white solid (20 mg, yield: 49%). LC-MS 404 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (DMSO-de, 400 MHz) δ 9.27 (s, 1 H), 8.80 (s, 1 H), 8.51 (s, 1 H), 8.49 (s, 1 H), 8.02-8.00 (dd, Ji = 1.6 Hz, J₂ = 8.4 Hz, 1 H), 7.46 (d, J = 8.8 Hz, 1 H), 7.03 (d, J = 8.0 Hz, 1 H), 6.67 (s, 1 H), 6.58-6.55 (dd, Ji = 2.4 Hz, J₂ = 8.4 Hz, 1 H), 5.90-5.85 (m, 1 H), 2.96- 2.93 (m, 1 H), 2.82-2.77 (m, 1 H), 2.51-2.47 (m, 1 H), 2.07-2.01 (m, 1 H).

Example 92: Preparation of compound A29-034, A29-046 and A29-047

A29-034 A29-046 A29-047

To a solution of 5-hydroxy-2,3-dihydroinden-l-one (1.48 g, 10.0 mmol) in dry DMF (20 mL) were added iodomethane (7.1 g, 50.0 mmol) and K₂C0₃ (2.76 g, 20.0 mmol). The mixture was stirred at room temperature for 20 h under N₂ atmosphere. The resulting mixture was quenched with water (100 mL), extracted with EtOAc (2 x 100 mL) and washed with brine (50 mL). The combined organic layer was dried over anhydrous Na₂S0₄ and evaporated to give the crude product SP-0010529-138 as a brown solid (1.9 g of the crude). LC-MS 163 (M+H)⁺, purity 95% (UV 214 nm). To a suspension of potassium tert-butanolate (291 mg, 2.6 mmol) in dry ethyl ether (20 mL) at 0 °C was added compound SP-0010529-118 (324 mg, 2.0 mmol) dropwise. The mixture was stirred at 0 °C for 0.5 h under stirring. Then the resulting mixture was added t- butyl nitrite (267 mg, 2.6 mmol) slowly and the reaction mixture was stirred for 1 h. The mixture was poured into cooled water (30 mL) and EtOAc (100 mL) was added to extract the product. The combined organic layer was dried over anhydrous Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 3:1) to give the desired compound SP-0010529-154 (a mixture of two isomers) as a brown color solid (270 mg, yield: 70%). LC-MS 192 (M+H)⁺, purity 36% (UV 214 nm).

To a solution of compound SP-0010529-154 (573 mg, 3.0 mmol) in MeOH / AcOH (30 mL / 2.0 mL) were added Pd/C (100 mg, 10%) and (Boc)₂0 (1.3 g, 6.0 mmol). The mixture was stirred at room temperature overnight under H₂ atmosphere. The resulting mixture was filtered off and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 2: 1) to give the desired compound SP-0010529-158 (270 mg, yield: 33%) as a pale yellow solid. LC-MS 222 (M-56+H)⁺, purity 49% (UV 214 nm).

To a solution of compound SP-0010529-158 (277 mg, 1.0 mmol) in THF (20 mL) were added DMAP (244 mg, 2.0 mmol) and (Boc)₂0 (436 mg, 2.0 mmol). The mixture was stirred at room temperature overnight. The reation mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 20: 1 - 10: 1) to give the desired compound SP-0010529-161 (210 mg, yield: 57%) as a colorless sticky oil. LC-MS 222 (M-56-100+H)⁺, purity 98% (UV 214 nm).

To a solution of compound SP-0010529-161 (210 mg, 0.56 mmol) in MeOH (10 mL) was added Pd/C (100 mg, 10%>). The mixture was stirred at room temperature overnight under H₂ atmosphere. The resulting mixture was filtered through celite and the filtrate was concentrated under reduced pressure to give the crude product SP-0010529-164 as pale yellow oil (200 mg, crude) which was used in next step of reaction without further purification. LC-MS 208 (M-100-56+H)⁺, purity 91% (UV 214 nm).

To a solution of compound SP-0010529-164 (200 g of the crude) in HC1 / 1,4- dioxane (5.0 mL). The mixture was stirred at room temperature for 20 h. After reaction, all of the volatiles were removed under reduced pressure to give the residue. It was dissolved in water (lO mL) and EtOAc (5 mL) was added to extract the by-products. The aqueous phase was dried to give the desired compound SP-0010529-170 (100 mg of the crude) as a form of HC1 salt. LC-MS 164 (M+H)⁺, purity 45% (UV 214 nm).

To a solution of compound SP-0010529-170 (91 mg, 0.5 mmol) in i-PrOH (10 mL) were added 4-chloro-6-fluoroquinazoline (100 mg, 0.3 mmol) and TEA (0.5 mL). The mixture was stirred at 85 °C for 3.0 h. The resulting mixture was evaporated and the residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 5: 1 - 1 : 1) to give the title compound A29-034 as a white solid (120 mg, yield: 77%). LC-MS 310 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (DMSO-d₆, 400 MHz) δ 8.51 (s, 1 H), 8.28- 8.20 (m, [1+1] H), 7.78-7.65 (m, [1+1] H), 7.15 (d, J = 8.0 Hz, 1 H), 6.86 (d, J = 6.0 Hz, 1 H), 6.75-6.73 (m, 1 H), 5.01-54.98 (m, 1 H), 3.73 (s, 3 H), 3.37-3.27 (m, 2 H), 3.04-2.92 (m, 2 H).

To a solution of compound A29-034 (80 mg, 0.26 mmol) in DCM (10 mL) was added BBr₃ (323 mg, 1.3 mmol) slowly at -10 °C. The mixture was stirred at 0 °C for 1 h. The reaction was quenched with a saturated solution of NaHC0₃ (20 mL) and extracted with DCM (100 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give the compound A29- 046 as a white solid (70 mg, yield: 91%). LC-MS 296 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (DMSO-de, 400 MHz) δ 9.18 (s, 1 H), 8.49 (s, 1 H), 8.33-8.27 (m, [1+1] H), 7.77- 7.73 (m, 1 H), 7.69-7.66 (dd, Ji = 2.8 Hz, J₂ = 8.8 Hz, 1 H), 7.02 (d, J = 8.0 Hz, 1 H), 6.66 (s, 1 H), 6.59-6.56 (dd, Ji = 2.4 Hz, J₂ = 8.4 Hz, 1 H), 4.98-4.96 (m, 1 H), 3.27-3.16 (m, 2 H), 2.99-2.93 (m, 2 H).

To a solution of compound A29-046 (50 mg, 0.17 mmol) and 4-(2-chloroethyl) morpholine hydrochloride (72 mg, 0.34 mmol) in DMF (4 mL) were added TBAI (12 mg, 0.03 mmol) and K₂CO₃(70 mg, 0.51 mmol). The mixture was stirred at 90 °C for 3 h. The reaction was cooled down, diluted with water (30 mL) and extracted with EtOAc (2 x 50 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 5: 1 - 2: 1) to give the title compound A29-047 as a white solid (18 mg, yield: 26%). LC-MS 409 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (DMSO- ₆, 400 MHz) δ 8.52 (s, 1 H), 8.36-8.34 (m, 1 H), 8.28-8.25 (m, 1 H), 7.79-7.75 (m, 1 H), 7.71-7.66 (m, 1 H), 7.17 (d, J = 8.4 Hz, 1 H), 6.90 (s, 1 H), 6.79-6.77 (dd, Ji = 2.4 Hz, J₂ = 8.0 Hz, 1 H), 5.01-4.98 (m, 1 H), 4.21-4.19 (m, 2 H), 3.74-3.70 (m, [2+2] H), 3.37-3.27 (m, [1+2] H), 3.05-2.94 (m, [2+2+2+1] H).

Example 93: Preparation of compound A29-048, A29-057 and A29-058

To a solution of 5-methoxy-2,3-dihydro-lH-inden-2-amine hydrochloride (290 mg, 1.0 mmol) in i-PrOH (15 mL) were added 4-chloro-6-iodoquinazoline (160 mg, 0.8 mmol) and TEA (1.0 mL). The mixture was stirred at 80 °C for 3 h. All of the volatiles were evaporated and the residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 2: 1) to give the desired compound A29-048 as a pale yellow solid (170 mg, yield: 50%). LC-MS 417 (M+H)⁺, purity 97% (UV 214 nm); 1H NMR (DMSO-de, 400 MHz) δ 8.78 (s, 1 H), 8.52 (s, 1 H), 8.43 (d, J = 6.4 Hz, 1 H), 8.42- 8.00 (dd, Ji = 1.6 Hz, J₂ = 8.4 Hz, 1 H), 7.46 (d, J = 8.0 Hz, 1 H), 6.85 (s, 1 H), 6.75-6.73 (dd, Ji = 2.4 Hz, J₂ = 8.4 Hz, 1 H), 5.01-4.97 (m, 1 H), 3.73 (s, 3 H), 3.36-3.26 (m, 2 H), 3.04-2.92 (m, 2 H).

To a solution of compound A29-048 (167 mg, 0.4 mmol) in DCM (20 mL) was added BBr₃ (500 mg, 2.0 mmol) slowly at -10 °C. The mixture was stirred at 0 °C for 1 h. The reaction was quenched with a saturated solution of NaHC0₃ (20 mL) and extracted with DCM (100 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give the title compound A29-057 as a white solid (140 mg, yield: 86%). LC-MS 404 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (DMSO-d₆, 400 MHz) δ 9.13 (s, 1 H), 8.79 (s, 1 H), 8.52 (s, 1 H), 8.41 (d, J = 6.4 Hz, 1 H), 8.02-8.00 (dd, Ji = 2.0 Hz, J₂ = 8.8 Hz, 1 H), 7.46 (d, J = 8.8 Hz, 1 H), 7.02 (d, J = 8.4 Hz, 1 H), 6.65 (s, 1 H), 6.58-6.56 (dd, Ji = 2.0 Hz, J₂ = 8.0 Hz, 1 H), 4.98-4.94 (m, 1 H), 3.31-3.21 (m, 2 H), 2.97-2.88 (m, 2 H). To a solution of 2 compound A29-057 (129 mg, 0.3 mmol) and 4-(2-chloroethyl) morpholine hydrochloride (129 mg, 0.6 mmol) in DMF (10 mL) were added TBAI (22.0 mg, 0.06 mmol) and K₂C0₃(124 mg, 0.9 mmol). The mixture was stirred at 90 °C for 3 h. The reaction was diluted with water (40 mL) and extracted with EtOAc (2 x 60 mL). The combined organic phase was evaporated and the residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 5: 1 - 2: 1) to give the title compound A29-058 as a white solid (40 mg, yield: 25%). LC-MS 517 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (CDC1₃, 400 MHz) δ 8.61 (s, 1 H), 7.94 (s, 1 H), 7.87-7.84 (m, 1 H), 7.47 (d, J = 8.8 Hz, 1 H), 7.08 (d, J = 4.4 Hz, 1 H), 6.75 (s, 1 H), 6.74-6.69 (m, 1 H), 5.96-5.93 (m, 1 H), 5.08-5.05 (m, 1 H), 4.04-4.01 (m, 2 H), 3.67-3.64 (m, [2+2] H), 3.42-3.33 (m, 2 H), 2.91-2.87 (m, 2 H), 2.84-2.83 (m, 2 H), 2.61-2.50 (m, [2+2] H).

Example 94: Preparation of compound A29-049, A29-056 and A29-064

A29-056 A29-064

To a solution of A29-048 (125 mg, 0.30 mmol) in dry DMA (4.0 mL) was added CuCN (138 mg, 1.5 mmol). The mixture was stirred at 1 10 °C for 20 h under N₂ atmosphere. The resulting mixture was quenched by water (20 mL), extracted with EtOAc (3 x 30 mL) and washed with brine (20 mL). The combined organic layer was dried over anhydrous Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (using petroleum ether: EtOAc = 10:1 - 2:1) to give the title compound A29-049 as a white solid (65 mg, yield: 68%). LC-MS 317 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (DMSO- ₆, 400 MHz) δ 8.98 (d, J = 1.6 Hz, 1 H), 8.69-8.62 (m, [1+1] H), 8.08-8.05 (dd, Ji = 2.0 Hz, J₂ = 8.8 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.16 (d, J = 8.4 Hz, 1 H), 6.87 (s, 1 H), 6.76-6.73 (dd, Ji = 2.4 Hz, J₂ = 8.0 Hz, 1 H), 5.02-5.00 (m, 1 H), 3.74 (s, 3 H), 3.38-3.28 (m, 2 H), 3.05-2.93 (m, 2 H). To a solution of A29-049 (40 mg, 0.12 mmol) in DCM (10 mL) was added BBr₃ (158 mg, 0.63 mmol) slowly at -10 °C. The mixture was stirred at 0 °C for 1 h. The reaction was quenched with a saturated solution of NaHC0₃ (20 mL) and extracted with DCM (100 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give the title compound A29-056 as a white solid (20 mg, yield: 55%). LC-MS 303 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (DMSC /e, 400 MHz) δ 9.15 (s, 1 H), 8.99 (s, 1 H), 8.65-8.61 (m, 1 H), 8.61 (s, 1 H), 8.07- 8.05 (dd, Ji = 2.0 Hz, J₂ = 8.8 Hz, 1 H), 7.78 (d, J = 8.4 Hz, 1 H), 6.67 (s, 1 H), 6.59-6.57 (dd, Ji = 2.4 Hz, J₂ = 8.0 Hz, 1 H), 5.00-4.96 (m, 1 H), 3.31-3.23 (m, 2 H), 2.98-2.45 (m, 2 H).

To a solution of compound A29-056 (103 mg, 0.3 mmol) and 4-(2-chloroethyl) morpholine hydrochloride (129 mg, 0.6 mmol) in DMF (10 mL) were added TBAI (22.0 mg, 0.06 mmol) and K₂C0₃ (124 mg, 0.9 mmol). The mixture was stirred at 90 °C for 3 h. The reaction was diluted with water (30 mL) and extracted with EtOAc (2 x 60 mL). The organic phase combined was evaporated and the residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 5: 1 - 2: 1) to give the title compound A29-064 as a white solid (45 mg, yield: 54%). LC-MS 416 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (CDC1₃, 400 MHz) δ 8.98 (s, 1 H), 8.66 (d, J = 6.4, 1 H), 8.62 (s, 1 H), 8.08- 8.05 (dd, Ji = 1.2 Hz, J₂ = 8.4 Hz, 1 H), 7.79 (d, J = 8.4 Hz, 1 H), 7.15 (d, J = 8.4 Hz, 1 H), 6.87 (s, 1 H), 6.76-6.74 (m, 1 H), 5.02-5.00 (m, 1 H), 4.08-4.04 (m, 2 H), 3.59-3.55 (m, [2+2] H), 3.38-3.28 (m, 2 H), 3.04-2.93 (m, 2 H), 2.69-2.67 (m, 2 H), 2.52-2.46 (m, [2+2] H).

Example 95: Preparation of compound A29-042

To a solution of A23-001 (100 mg, 0.33 mmol) in DMF (2.0 mL) were added 2- phenoxyacetic acid (100 mg, 0.66 mmoL), HATU (380 mg, 1.00 mmol) and triethylamine (0.24 mL, 1.65 mmol). The reaction mixture was stirred at 50 °C for 8 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-042 as a white solid (30 mg, yield: 21%). LC-MS 436 (M+H) ⁺, purity 95% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 10.02 (s, 1 H), 8.97 (s, 1 H), 8.67 (d, J = 6.8 Hz, 1 H), 8.63 (s, 1H), 8.06 (d, J = 8.4 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.62 (s, 1 H), 7.41 (d, J = 8.4 Hz, 1 H), 7.32 (t, J = 6.8 Hz, 2 H), 7.21 (d, J = 8.0 Hz, 1 H), 7.01-6.96 (m, 3 H), 5.03-4.99 (m, 1 H), ), 3.12 (s, 2 H), 3.41-3.39 (m, 2 H), 3.06-2.98 (m, 2 H), 2.62-2.50 (m, 4 H), 1.02(t, J = 6.8 Hz, 6 H).

Example 96: Preparation of compound A29-04

To a solution of A23-001 (100 mg, 0.33 mmol) in DMF (2.0 mL) were added 2- (ethylthio) acetic acid (79 mg, 0.66 mmoL), HATU (380 mg, 1.00 mmol) and triethylamine (0.24 mL, 1.65 mmol). The reaction mixture was stirred at 50 °C for 8 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-043 as a white solid (30 mg, yield: 23%). LC-MS 404 (M+H) ⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 10.00 (s, 1 H), 8.98 (s, 1 H), 8.66 (d, J = 6.8 Hz, 1 H), 8.63 (s, 1 H), 8.07 (d, J = 8.8 Hz, 1 H), 7.79 (d, J = 8.4 Hz, 1 H), 7.58 (s, 1 H), 7.33 (d, J = 8.0 Hz, 1 H), 7.19 (d, J = 8.0 Hz, 1 H), 7.01-6.96 (m, 3 H), 5.04-4.99 (m, 1 H), ), 3.39-3.32 (m, 2 H), 3.30 (s, 2 H), 3.06-2.96 (m, 2 H), 2.66-2.60 (m, 2 H), 1.19 (t, J = 6.0 Hz, 3 H).

Example 97: Preparation of A29-044

To a solution of A29-043 (100 mg, 0.25 mmol) in THF (10 mL) was added 3- chlorobenzoperoxoic acid (86 mg, 0.50 mmoL). The mixture was stirred at room temperature for 1 h. THF was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-044 as a white solid (21 mg, yield: 19%). LC-MS 436 (M+H)⁺, purity 96% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.66 (s, 1 H), 8.49 (s, 1 H), 7.89 (d, J = 8.8 Hz, 1 H), 7.70 (d, J = 8.8 Hz, 1 H), 7.45 (s, 1 H), 7.23 (d, J = 8.0 Hz, 1 H), 7.12 (d, J = 8.0 Hz, 1 H), 5.07-5.04 (m, 1 H), 3.38-3.22 (m, 4 H), 3.02-2.97 (m, 2 H), 1.30 (t, J = 7.6 Hz, 3 H).

Example 98: Preparation of compound A29-045

To a solution of A23-001 (150 mg, 0.50 mmol) in DCM (10 mL) were added 2- chloroacetyl chloride (84 mg, 0.75 mmoL), triethylamine (0.20 mL, 1.50 mol). The mixture was stirred at room temperature for 8 h. DCM was removed by rotary evaporation. The resulting residue was taken up into ethyl ether (150 mL), washed with saturated NaHC0₃ (50 mL).The organic extracts were dried over Na₂S0₄, filtered and concentrated to yield a crude product SP-0011507-192 which was used in next step of reaction without further purification (180 mg, yield: 96%).

To a solution of SP-0011507-192 (180 mg, 0.47 mmol) in DMF (2.0 mL) was added KI (8.30 mg, 0.05 mmol). The mixture was stirred at 120 °C under microwave irradiation for 1 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-045 as a white solid (55 mg, yield: 29%). LC-MS 410 (M+H) ⁺, purity 94% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 10.25 (s, 1 H), 8.97 (s, 1 H), 8.67 (d, J = 6.4 Hz, 1 H), 8.62 (s, 1 H), 8.07 (d, J = 8.4 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.71 (s, 1 H), 7.55 (s, 1 H), 7.36 (d, J = 8.0 Hz, 1 H), 7.22-7.18 (m, 2 H), 6.94 (s, 1 H), 5.04-5.00 (m, 1 H), 4.90 (s, 2 H), 3.41-3.35 (m, 2 H), 3.05-2.97 (m, 2 H).

Example 99: Preparation of A29-052

To a solution of A23-001 (150 mg, 0.50 mmol) in DMF (2.0 mL) were added 1- hydroxycyclopropanecarboxylic acid (102 mg, 1.00 mmoL), HATU (570 mg, 1.50 mmol) and triethylamine (0.36 mL, 2.50 mmol). The reaction mixture was stirred at 50 °C for 8 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-052 as a white solid (70 mg, yield: 36%). LC-MS 386 (M+H) ⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.68 (s, 1 H), 8.98 (s, 1 H), 8.68 (d, J = 6.8 Hz, 1 H), 8.63 (s, 1 H), 8.07 (d, J = 8.8 Hz, 1 H), 7.79 (d, J = 8.4 Hz, 1 H), 7.70 (s, 1 H), 7.50 (d, J = 8.0 Hz, 1 H), 7.18 (d, J = 8.4 Hz, 1 H), 6.54 (s, 1 H), 5.04-5.00 (m, 1 H), 3.39-3.35 (m, 2 H), 3.05-2.97 (m, 2 H), 1.15-1.12 (m, 2 H), 0.97-0.93 (m, 1 H).

Example 100: Preparation of compound A29-059, A29-075 and A29-081

To a solution of A29-048 (417 mg, 1.0 mmol) in dry DMSO (10.0 mL) were added Cul (19 mg, 0.1 mmol) and sodium methanesulfmate (590 mg, 5.0 mmol). The mixture was stirred at 110 °C for 20 h under N₂ atmosphere. The resulting mixture was quenched by water (50 mL), extracted with EtOAc (2 x 100 mL) and washed with brine (40 mL). The combined organic layer was dried over anhydrous Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography with (using petroleum ether : EtOAc = 10: 1 - 1 : 1) to give the title compound A29-059 (250 mg, yield: 67%) as a white solid. LC-MS 370 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (DMSO-de, 400 MHz) δ 9.11 (s, 1 H), 9.00-8.98 (m, 1 H), 8.22-7.90 (m, [1+1] H), 7.16 (d, J = 8.0 Hz , 1 H), 6.86 (s, 1 H), 6.76-6.74 (m, 1 H), 5.07-5.04 (m, 1 H), 3.73 (s, 3 H), 3.39- 3.30 (m, 2 H), 3.28 (s, 3 H), 3.09-2.97 (m, 2 H).

To a solution of compound A29-059 (50 mg, 0.13 mmol) in DCM (10 mL) was added BBr₃ (162 mg, 0.65 mmol) slowly at -10 °C. The mixture was stirred at 0 °C for 2 h. The reaction was quenched with a saturated solution of NaHC0₃ (20 mL) and extracted with DCM (100 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give the title compound A29-075 as a yellow color solid (35 mg, yield: 76%). LC-MS 356 (M+H)⁺, purity 99% (UV 214 nm). 1H NMR (DMSO-d₆, 400 MHz) δ 9.15 (s, 1 H), 9.04 (s, 1 H), 8.92 (d, J = 6.4 Hz ,

1 H), 8.62 (s, 1 H), 8.20-8.17 (dd, Ji = 2.0 Hz, J₂ = 8.8 Hz, 1 H), 7.86 (d, J = 8.8 Hz, 1 H),

7.03 (d, J = 8.0 Hz, 1 H), 6.66 (s, 1 H), 6.59-6.56 (dd, Ji = 2.0 Hz, J₂ = 8.0 Hz, 1 H), 5.05- 5.02 (m, 1 H), 3.31-3.23 (m, [2+3] H), 3.03-2.92 (m, 2 H).

To a solution of 2 compound A29-075 (210 mg of the crude) and 4-(2-chloroethyl) morpholine hydrochloride (64 mg, 0.30 mmol) in DMF (10 mL) were added TBAI (37 mg, 0.1 mmol) and K2C03(414 mg, 3.0 mmol). The mixture was stirred at 90 °C for 3 h. The reaction was diluted with water (50 mL) and extracted with EtOAc (2 x 100 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (using petroleum ether: EtOAc = 5: 1 - 2: 1) to give the title compound A29-081 as a white solid (20 mg). LC-MS 469 (M+H)⁺, purity 100% (UV 214 nm). 1H NMR (DMSO-d₆, 400 MHz) δ 9.03 (s, 1 H), 8.93 (d, J = 6.4 Hz, 1 H), 8.63 (s, 1 H), 8.20-8.17 (dd, Ji = 2.0 Hz, J₂ = 8.8 Hz, 1 H), 7.98 (d, J = 8.8 Hz, 1 H), 7.15 (d, J = 8.4 Hz, 1 H), 6.87 (s, 1 H), 6.76-6.73 (dd, Ji = 2.0 Hz, J₂ =

8.4 Hz, 1 H), 5.07-5.03 (m, 1 H), 4.07-4.04 (m, 2 H), 3.59-3.55 (m, [2+2] H), 3.38-3.30 (m,

2 H), 3.27 (s, 3 H), 3.08-2.97 (m, 2 H), 2.68-2.65 (m, 2 H), 2.51-2.45 (m, [2+2] H).

Example 101: Preparation of compound A29-062

To a solution of A23-001 (200 mg, 0.66 mmol) in DCM (20 mL) were added acetic anhydride (135 mg, 1.32 mmoL), 4-dimethylamiopryidine (24.0 mg, 0.20 mmoL) and triethylamine ( 0.48 mL, 3.30 mol). The mixture was stirred at room temperature for 6 h. DCM was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-062 as a white solid (120 mg, yield: 53%). LC-MS 344 (M+H)⁺, purity 96% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.86 (s, 1 H), 8.97 (s, 1 H), 8.66 (d, J = 6.4 Hz, 1 H), 8.62 (s, 1 H), 8.06 (d, J = 8.4 Hz, 1 H), 7.78 (d, J = 8.8 Hz, 1 H), 7.57 (s, 1 H), 7.33 (d, J = 8.0 Hz, 1 H), 7.17 (d, J = 8.0 Hz, 1 H), 5.04-4.99 (m, 1 H), 3.40-3.36 (m, 2 H), 3.05-2.95 (m, 2 H), 2.03 (s, 3 H). Example 102: Preparation of compound A29-063

To a solution of compound SP-0011507-011 (1.00 g, 5.62 mmol) and Boc₂0 (1.83 g, 8.43 mmol) in dioxane\H₂0 (100\25 mL) was added NaHC0₃ (944 mg, 11.2 mmol). The mixture was stirred at room temperature for 8 h. The resulting residue was taken up into ethyl ether (200 mL), washed with water (50 mL). The organic extracts were dried over Na₂SC"4, filtered and concentrated under rerduced pressure to give crude product. The crude product was purified by silica gel column chromatography (using petroleum ether/EtOAc = 20: 1 - 9: 1) to give compound SP-0011507-017 as a white solid (1.25 g, yield: 80%). LC- MS 223 (M-55+H)⁺, purity 95% (UV 214 nm).

A mixture of SP-0011507-017 (380 mg, 1.37 mmol) and Pd/C (38 mg, 10%) in MeOH (50 mL) was stirred at room temperature for 3 h under H₂ atmosphere. The resulting mixture was filtered off and the filtrate was concentrated to give the desired SP-0011507- 023 (330 mg, yield: 97%). LC-MS 271 (M+Na)⁺, purity 87% (UV 214 nm).

To a solution of compound SP-0011507-023 (330 mg, 1.33 mmol) in isopropanol (30 mL) were added 4-chloro-6-iodoquinazoline (464 mg, 1.60 mmol) and triethylamine (057 mL, 3.99 mmol). The mixture was stirred at 70 °C for 9 h. The mixture was cooled down and isopropanol was removed by rotary evaporation. The residue was purified by silica gel column chromatography (using petroleum ether/EtOAc = 6:1-1 :2) to give compound SP-0011507-027 as a yellow solid (310 mg, yield: 46%). LC-MS 503 (M+H)⁺, purity 60% (UV 214 nm).

To a solution compound SP-0011507-027 (310 mg, 0.62 mmol) in DCM (10 mL) was added a solution of HCl in 1,4-dioxane (4 M, 5 mL). The mixture was stirred at room temperature for 8 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-063 as a white solid ( 80 mg, yield: 32%). LC-MS 403 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.78 (s, 1 H), 8.99 (s, 1 H), 8.56 (s, 1 H), 8.09 (d, J = 8.8 Hz, 1 H), 7.66 (s, 1 H), 7.53 (d, J = 9.2 Hz, 1 H), 7.49 (d, J = 8.4 Hz, 1 H), 7.19 (d, J = 8.0 Hz, 1 H), 3.75-3.69 (m, 1 H), 3.24-3.13 (m, 2 H), 3.11-2.99

(m, 2 H), 2.63-2.53 (m, 2 H).

Example 103: Preparation of compound A29-067

SP-0011507-034 A29-067

A mixture of 2-amino-4-chlorobenzoic acid (6.00 g, 35.1 mmol) in formamide (23.7 g, 530 mmol) was heated at 200 °C for 6 h. The reaction mixture was cooled down, filtered, washed with water (5 x 50 mL), dried under reduced pressure to afford crude product SP- 0011507-028 (4.60 g, yield: 73%). The resulting crude product was used in the next step of reaction without further purification.

The crude product SP-0011507-028 (4.60 g, 25.5 mmol) was mixed with thionyl dichloride (29.5 g, 0.25 mol), and catalytic amount of anhydrous DMF (1.00 mL). The resulting mixture was heated to reflux for 24 h. The reaction mixture was cooled down and excess thionyl dichloride was removed by rotary evaporation. DCM (10 mL) was added to dissolve the solid, and then resulting crude product SP-0011507-032 (4.00 g, yield: 80%) was used in the next step of reaction without further purification.

To a solution of compound SP-0011507-032 (2.00 g, 10.1 mmol) and 5-nitro-2,3- dihydro-lH-inden-2-amine (1.50 g, 8.42 mmol) in isopropyl alcohol (100 mL) was added triethylamine (3.6 mL, 25.3 mmol). The resulting solution was heated to 70 °C for 9 h. The mixture was cooled down and excess of isopropyl alcohol was removed by rotary evaporation. The residue was purified by silica gel column chromatography (using petroleum ether/EtOAc = 4: 1-1 :2) to give compound SP-0011507-034 as a yellow solid (1.24 g, yield: 44%). LC-MS 341 (M+H)⁺, purity 83% (UV 214 nm).

A mixture of SP-0011507-034 (1.20 g, 3.53 mmol) and Pd/C (120 mg, 10%) in MeOH (100 mL) and THF (50 mL) was stirred at room temperature for 12 h under ¾ atmosphere. The resulting mixture was filtered off and the filtrate was concentrated to give the crude product. The residue was purified by Prep-HPLC to give A29-067 as a white solid (1.00 g, yield: 92%). LC-MS 311 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.50 (s, 1 H), 8.43 (d, J = 6.8 Hz, 1 H), 8.40 (d, J = 8.0 Hz, 1 H), 7.71 (s, 1 H), 7.62 (s, 1 H), 7.55 (d, J = 8.4 Hz, 1 H), 6.88 (d, J = 8.0 Hz, 1 H), 6.46 (s, 1 H), 6.39 (d, J = 8.4 Hz, 1 H), 4.96-4.93 (m, 1 H), 4.85 (s, 2 H), 3.23-3.15 (m, 2 H), 2.92-2.83 (m, 2 H). Example 104: Preparation of compound A29-068

A29-067 50 °C, 8 h A29-068

To a solution of A29-067(120 mg, 0.39 mmol) in DMF (2.0 mL) were added 2- hydroxyacetic acid (59 mg, 0.78 mmoL), HATU (445 mg, 1.17 mmol) and triethylamine (0.28 mL, 1.95 mmol). The reaction mixture was stirred at 50 °C for 8 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-068 as a white solid (21 mg, yield: 15%). LC-MS 369 (M+H) ⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.56 (s, 1 H), 8.53 (s, 1 H), 8.48 (d, J = 6.4 Hz, 1 H), 8.39 (d, J = 8.8 Hz, 1 H), 7.73 (s, 1 H), 7.64 (s, 1 H), 7.56 (d, J = 8.8 Hz, 1 H), 7.45 (d, J = 8.4 Hz, 1 H), 7.17 (d, J = 8.4 Hz, 1 H), 5.66 (s, 1 H), ), 5.04-4.98 (m, 1 H), 3.97 (s, 1 H), 3.43-3.33 (m, 2 H), 3.06-2.96 (m, 2 H).

Example 105: Preparation of compound A29-069

A29-067 A29-069

To a solution of A29-067(120 mg, 0.39 mmol) in DCM (20 mL) was added acetic anhydride (80 mg, 0.78 mmoL), 4-dimethylamiopryidine (4.8 mg, 0.04 mmoL) and triethylamine (0.17 mL, 1.17 mol). The mixture was stirred at room temperature for 6 h. DCM was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-069 as a white solid (29 mg, yield: 21%). LC-MS 353 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.84 (s, 1 H), 8.52 (s, 1 H), 8.47 (d, J = 6.8 Hz, 1 H), 8.39 (d, J = 8.8 Hz, 1 H), 7.72 (s, 1 H), 7.55 (d, J = 8.8 Hz, 1 H), 7.31 (d, J = 8.4 Hz, 1 H), 7.15 (d, J = 8.4 Hz, 1 H), 5.03-4.98 (m, 1 H), 3.36-3.30 (m, 2 H), 3.05-2.95 (m, 2 H), 2.03 (s, 3 H).

Example 106: Preparation of compound A29-071

-071

To a solution of A29-067(120 mg, 0.39 mmol) in DMF (2.0 mL) was added 2- morpholinoacetic acid (113 mg, 0.78 mmoL), HATU (445 mg, 1.17 mmol), triethylamine (0.28 mL, 1.95 mmol). The reaction mixture was stirred at 50 °C for 8 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-071 as a white solid (27 mg, yield: 16%). LC-MS 438 (M+H) ⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.65 (s, 1 H), 8.52 (s, 1 H), 8.47 (d, J = 6.8 Hz, 1 H), 8.39 (d, J = 9.2 Hz, 1 H), 7.73 (s, 1 H), 7.64 (s, 1 H), 7.56 (d, J = 10. Hz, 1 H), 7.39 (d, J = 8.0 Hz, 1 H), 7.18 (d, J = 8.0 Hz, 1 H), 5.04-4.98 (m, 1 H), 3.63 (t, J = 4.4 Hz, 4 H), 3.37-3.30(m, 4 H), 3.29-3.28 (m, 2 H), 3.10 (s, 2 H), 3.06-2.95 (m, 2 H).

Example 107: Preparation of compound A29-072

To a solution of A23-001 (150 mg, 0.50 mmol) in MeOH (10 mL) was added acetaldehyde (44.0 mg, 1.00 mmoL).The mixture was stirred at room temperature for 3 h. NaCNBH₃ (63.0 mg, 1.00 mmoL) was added. The resulting mixture was stirring at room temperature for 24 h. MeOH was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-072 as a white solid (47 mg, yield: 26%). LC-MS 358 (M+H)⁺, purity 95% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.00 (s, 1 H), 8.65 (d, J = 6.4 Hz, 1 H), 8.61 (s, 1 H), 8.06 (d, J = 8.8 Hz, 1 H), 7.78 (d, J = 8.8 Hz, 1 H), 7.57 (s, 1 H), 7.03 (d, J = 8.4 Hz, 1 H), 6.59 (s, 1 H), 6.51 (d, J = 7.2 Hz, 1 H), 5.00-4.95 (m, 1 H), 3.33- 3.27 (m, 4 H), 3.26-3.22 (m, 2 H), 3.00-2.87 (m, 2 H), 1.07 (t, J = 6.8 Hz, 6 H). Example 108: Preparation of compound A29-073

SP-0011507-011

A mixture of SP-0011507-011 (600 mg, 3.37 mmol) and Pd/C (60 mg, 10%) in MeOH (60 mL) was stirred at room temperature for 4 h under ¾ atmosphere. The resulting mixture was filtered off and the filtrate was concentrated to give the crude product SP- 0011507-054 (400 mg, yield: 80%). The resulting crude product was used in the next step of reaction without further purification.

To a solution of compound SP-0011507-054 (400 mg, 2.70 mmol) and 4,6- dichloroquinazoline (642 mg, 3.24 mmol) in isopropyl alcohol (50 mL) was added triethylamine (1.17 mL, 8.10 mmol). The resulting solution was heated to 50 °C for 9 h. The mixture was cooled down and excess of isopropyl alcohol was removed by rotary evaporation. The residue was purified by silica gel column chromatography (using petroleum ether/EtOAc = 3: 1-1 :2) to give compound A29-073 as a yellow solid (500 mg, yield: 60%). LC-MS 311 (M+H)⁺, purity 96% (UV 214 nm); 1H NMR (400 MHz, DMSO- d6) δ 8.54 (s, 1 H), 8.52 (s, 1 H), 8.39 (d, J = 6.8 Hz, 1 H), 7.77 (d, J = 8.8 Hz, 1 H), 7.69 (d, J = 8.8 Hz, 1 H), 6.89 (d, J = 8.0 Hz, 1 H), 6.47 (s, 1 H), 6.40 (d, J = 8.0 Hz, 1 H), 4.96- 4.91 (m, 1 H), 4.88 (s, 2 H), 3.25-3.16 (m, 2 H), 2.92-2.83 (m, 2 H).

Example 109: Preparation of compound A29-074

SP-0011507-011 70 °C, 9 h SP-0011507-047

SP-0011507-051 r.t., 6 h To a solution of compound SP-0011507-011 (800 mg, 4.49 mmol) and 4,6- dichloroquinazoline (1.06 g, 5.39 mmol) in isopropyl alcohol (100 mL) was added triethylamine (1.94 mL, 13.5 mmol). The resulting solution was heated to 70 °C for 9 h. The mixture was cooled down and excess of isopropyl alcohol was removed by rotary evaporation. The residue was purified by silica gel column chromatography (using petroleum ether/EtOAc = 3: 1-1 :2) to give compound SP-0011507-047 as a yellow solid (850 mg, yield: 56%). LC-MS 341 (M+H)⁺, purity 63% (UV 214 nm).

A mixture of SP-0011507-047 (850 mg, 2.50 mmol) and Pd/C (85 mg, 10%) in MeOH (100 mL) and THF (50 mL) was stirred at room temperature for 12 h under H₂ atmosphere. The resulting mixture was filtered off and the filtrate was concentrated to give SP-0011507-051 as a yellow solid (621 mg, yield: 90%>). The resulting crude product was used in the next step of reaction without further purification.

To a solution of compound SP-0011507-051 (150 mg, 0.54 mmol) in DCM (20 mL) was added acetic anhydride (104 mg, 1.08 mmoL), 4-dimethylamiopryidine (6.10 mg, 0.05 mmoL) and triethylamine (0.23 mL, 1.62 mol). The mixture was stirred at room temperature for 6 h. DCM was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-074 as a white solid (35 mg, yield: 20%). LC-MS 319 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.86 (s, 1 H), 8.52 (s, 2 H), 8.33 (t, J = 8.0 Hz, 2 H), 7.76 (t, J = 8.4 Hz, 1 H), 7.69 (d, J = 7.2 Hz, 1 H), 7.55 (s, 1 H), 7.49(t, d = 7.6 Hz, 1 H), 7.32 (d, J = 8.0 Hz, 1 H), 7.16 (d, J = 8.0 Hz, 1 H), 5.06-4.99 (m, 1 H), 3.35-3.28 (m, 2 H), 3.06-2.96 (m, 2 H), 2.03 (s, 3 H).

Example 110: Preparation of compound A29-076

To a solution of compound SP-0011321-028 (120 mg, 0.408 mmol) in DMF (3 mL) were added 2-hydroxyacetic acid (37 mg, 0.490 mmol), HATU (186 mg, 0.490 mmol) and triethylamine (412 mg, 4.08 mmol). The mixture was stirred at 40 °C for 5 h. The reaction mixture was poured into water (15 mL) and extracted with EtOAc (2 x 20 mL). The organic layer was washed with water (15 mL) and brine (20 mL), dried over Na₂S0₄, filtered, and concentrated. The residue was purified by Prep-HPLC to give A29-076 as a white solid (50 mg, yield: 35%). LC-MS 353 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.56 (s, 1 H), 8.51 (s, 1 H), 8.47-8.42 (m, 2 H), 7.65 (s, 1 H), 7.46-7.39 (m, 3 H), 7.17 (d, J = 8.0 Hz, 1 H), 5.66 (t, J = 5.6 Hz, 1 H), 5.05-4.99 (m, 1 H), 3.98 (d, J = 3.6 Hz, 2 H), 3.38-3.29 (m, 2 H), 3.00 (td, J = 16.0 Hz, 6.0 Hz, 2 H).

Example 111: Preparation of compound A29-077

To a solution of compound SP-0011321-028 (120 mg, 0.408 mmol) in DMF (3 mL) was added 3-hydroxy-3-methylbutanoic acid (96 mg, 0.816 mmol), HATU (310 mg, 0.816 mmol) and triethylamine (410 mg, 0.408 mmol). The mixture was stirred at 40 °C for 16 h. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (2 x 20 mL). The organic layer was washed with water (15 mL) and brine (15 mL), dried over Na₂S0₄, filtered, and concentrated. The residue was purified by Prep-HPLC to give A29- 077 as a white solid (40 mg, yield: 20%). LC-MS 395 (M+H)⁺, purity 97% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.77 (s, 1 H), 8.51 (s, 1 H), 8.47-8.41 (m, 2 H), 7.57 (s, 1 H), 7.44-7.39 (m, 2 H), 7.32 (d, J = 8.0 Hz, 1 H), 7.16 (d, J = 8.0 Hz, 1 H), 5.05-4.99 (m, 1 H), 4.76 (s, 1 H), 3.37-3.28 (m, 2 H), 3.00 (td, J = 17.2 Hz, 6.4 Hz, 2 H), 2.41 (s, 2 H), 1.22 (s, 6 H).

Example 112: Preparation of compound A29-078

A02-171 A29-078

To a solution of compound A02-171 (150 mg, 0.510 mol) in DMF (4 mL) was added biotin (150 mg, 0.612 mmol), HATU (233 mg, 0.612 mmol) and triethylamine (515 mg, 5.10 mmol). The mixture was stirred at 40 °C for 5 h. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (2 x 30 mL). The organic layer was washed with water (15 mL) and brine (15 mL), dried over Na₂S0₄, filtered, and concentrated. The residue was purified by Prep-HPLC to give A29-078 as a white solid (110 mg, yield: 41%). LC-MS 521 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.80 (s, 1 H), 8.51 (s, 1 H), 8.28-8.22 (m, 2 H), 7.77 (dd, J = 9.2 Hz, 6.4 Hz, 1 H), 7.67 (td, J = 8.8 Hz, 2.8 Hz, 1 H), 7.58 (s, 1 H), 7.33 (d, J = 8.0 Hz, 1 H), 7.17 (d, J = 8.0 Hz, 1 H), 6.45 (s, 1 H), 6.37 (s, 1 H), 5.03-4.97 (m, 1 H), 4.33-4.29 (m, 1 H), 4.16-4.13 (m, 1 H), 3.37-3.29 (m, 2 H), 3.15-3.10 (m, 1 H), 3.00 (td, J = 17.2 Hz, 6.0 Hz, 2 H), 2.83 (dd, J = 12.4 Hz, 5.2 Hz, 1 H), 2.58 (d, J = 12.4 Hz, 1 H), 2.30 (t, J = 7.6 Hz, 2 H), 1.66-1.34 (m, 6 H).

Example 113: Preparation of compound A29-079

To a solution of A23-001 (100 mg, 0.33 mmol) in DMF (2.0 mL) were added biotin (161 mg, 0.66 mmoL), HATU (376 mg, 0.99 mmol) and triethylamine (0.24 mL, 1.65 mmol). The reaction mixture was stirred at 50 °C for 8 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give SP-0011507-061 as a white solid (69 mg, yield: 40%). LC-MS 528 (M+H) ⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.81 (s, 1 H), 8.97 (s, 1 H), 8.66 (d, J = 6.4 Hz, 1 H), 8.62 (s, 1 H), 8.07 (d, J = 8.4 Hz, 1 H), 7.79 (d, J = 8.4 Hz, 1 H), 7.59 (s, 1 H), ), 7.33 (d, J = 8.8 Hz, 1 H), 7.17 (d, J = 8.4 Hz, 1 H), 6.44 (s, 1 H), 6.37 (s, 1 H), 5.04-5.00 (m, 1 H), 4.31 (t, J = 7.6 Hz, 1 H), 4.14 (t, J = 5.2 Hz, 1 H), 3.34-3.35 (m, 2 H), 3.14-3.10 (m, 1 H), 3.05-2.95 (m, 2 H), 2.58 ( d, J = 12.4 Hz, 1 H), 2.30 (t, J = 7.2 Hz, 2 H), 1.66-1.34 (m, 6 H).

Example 114: Preparation of compound A29-080

Ck .COOH ,S. ,COOH

1.NaS, NaOH, 60 °C, 2.5 |l ]^ Pd/C, H₂(g) Formamide ki -_MNn0„₂ 22..((CCHH,₃)),₂SS00₄₄,, NNaaOOHH,, 110000 °°CC,, 11 h ^ ^<i^ -_WHn0„₂ MMeeOOHH., RRTT., 1166 '^^fK~H\\₂ 200 °C, 5 h

SP-0011321 -011 SP-0011321-015

SP-0011321-017 SP-0011321 -026 SP-0011321-041

SP-0011321-042 A29-080

To a mixture of 5-chloro-2-nitrobenzoic acid (25.2 g, 0.125 mol) in water (50 mL) was added aqueous NaOH (2M, 42 mL, 0.084 mol) to completely dissolve 5-chloro-2- nitrobenzoic acid. A solution of NaS nonahydrate (33.0 g, 0.137 mmol) in water (75 mL) was added and the resulting mixture was stirred at 60 °C for 2.5 h. The resulting red solution was added to a mixture of aqueous NaOH (50%, 10 mL, 0.125 mol) and water (15 mL). Dimethyl sulfate (24 mL, 0.250 mol) was added and the reaction mixture was heated under reflux for 1 h. The reaction mixture was cooled down and acidified with aqueous HCl (5 M, 32 mL, 0.160 mmol). The yellow precipitate was filtered off, washed with water, dried at 60 °C under vacuum to afford product SP-0011321-011(22.5 g, yield: 90%). LC- MS 214 (M+H)⁺, purity 100% (UV 214 nm)

A mixture of SP-0011321-011 (22.5 g, 0.106 mol) and 10% Pd/C (1 g) in MeOH (150 mL) was stirred at room temperature overnight under H₂ atmosphere. The resulting mixture was filtered through celite and the filtrate was evaporated to afford SP-0011321- 015 (18.0 g, yield: 98%). LC-MS 184 (M+H)⁺, purity 100% (UV 214 nm)

A mixture of compound SP-0011321-015 (1.84g, 10.0 mmol) in formamide (9.00 g, 0.20 mol) was heated at 200 °C for 5 h. The reaction mixture was cooled down and poured into water (25 mL). The yellow precipitate was filtrated, washed with water (5 x 15 mL) and dried under vacuum to afford crude product SP-0011321-017 (1.63 g, yield: 85%). The resulting crude product was used in the next step of reaction without further purification.

A mixture of the crude product SP-0011321-017 (1.00 g, 5.21 mmol), thionyl dichloride (12.0 g, 0.1 mol) and catalytic amount of anhydrous DMF (0.5 mL) was heated to reflux for 4 h under N₂ atmosphere. The mixture was cooled down and excess thionyl dichloride was removed by rotary evaporation to give the crude product SP-0011321-026 (1 g). LC-MS 211 (M+H)⁺, purity 83% (UV 214 nm) . The crude product was used in the next step of rection without further purification.

A mixture of 2, 3-dihydro-lH-indene-2, 5-diamine (200 mg, 1.35 mmol), triethylamine (1.36 g, 13.5 mmol) in isopropyl alcohol (8.0 mL) and SP-0011321-026 (1 g, crude) was heated to 65 °C under N₂ for 2 h. The reaction mixture was cooled down and excess of isopropyl alcohol was removed by rotary evaporation. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 2: 1 - 2:2) to afford the compound SP-0011321-041 (435 mg, yield: 75%). LC-MS 325 (M+H)⁺, purity 93% (UV 214 nm).

To a solution of compound SP-0011321-041 (150 mg, 0.466 mmol) in DCM (6 mL) cooled in an ice-bath were added acetic anhydride (95 mg, 0.932 mmol), triethylamine (470 mg, 4.66 mmol) and DMAP (6 mg). The reaction mixture was stirred at room temperature for 2 h, and concentrated to give a crude product SP-0011321-042 which was used in next step of reaction without further purification (160 mg, yield: 94%>).

To a solution of compound SP-0011321-042 (160 mg, 0.466 mmol) in 1,4-dioxane (5 mL) was added a solution of Oxone (429 mg, 0.699 mmol) in water (3 mL). The reaction mixture was stirred at room temperature for 0.5 h and concentrated under reduced pressure. The residue was partitioned between EtOAc (30 mL) and brine (20 mL). The organic layer was separated and the aqueous phase was extracted with EtOAc (20 mL). The combined organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give the product A29-080 as a white solid (50 mg, yield: 27%). LC-MS 397 (M+H)⁺, purity 96% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.86 (s, 1 H), 9.03 (d, J = 2.0 Hz, 1 H), 8.94 (s, 1 H), 8.64 (s, 1 H), 8.19 (dd, J = 8.8 Hz, 2.0 Hz, 1 H), 7.87 (dd, J = 8.8 Hz, 5.6 Hz, 1 H), 7.57 (s, 1 H), 7.32 (d, J = 8.0 Hz, 1 H), 7.17 (d, J = 8.0 Hz, 1 H), 5.09-5.04 (m, 1 H), 3.38-3.30 (m, 2 H), 3.28 (s, 3 H), 3.03 (td, J = 16.8 Hz, 6.0 Hz, 2 H) , 2.03 (s, 3 H).

Example 115: Preparation of compound A29-082

r.t., 6 h

A29-073 A29-082 To a solution of A29-073(120 mg, 0.39 mmol) in DCM (20 mL) was added acetic anhydride (80 mg, 0.78 mmoL), 4-dimethylamiopryidine (4.8 mg, 0.04 mmoL) and triethylamine (0.17 mL, 1.17 mol). The reaction mixture was stirred at room temperature for 6 h. DCM was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-082 as a white solid (40 mg, yield: 29%). LC-MS 353 (M+H)⁺, purity 98% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.85 (s, 1 H), 8.53 (s, 2 H), 8.47 (d, J = 6.8 Hz, 1 H), 8.42 (d, J = 6.4 Hz, 1 H), 7.78 (d, J = 8.8 Hz, 1 H), 7.70 (d, J = 8.8 Hz, 1 H), 7.56 (s, 1H), 7.33 (d, J = 8.0 Hz, 1 H), 7.16 (d, J = 8.0 Hz, 1 H), 5.03-4.98 (m, 1 H), 3.37- 3.28 (m, 2 H), 3.05-2.95 (m, 2 H), 2.03 (s, 3 H).

Example 116: Preparation of compound A29-083

To a solution of SP-0011507-051 (200 mg, 0.72 mmol) in DMF (2.0 mL) were added 3-hydroxy-3-methylbutanoic acid (170 mg, 1.44 mmoL), HATU (684 mg, 1.80 mmol) and triethylamine (0.52 mL, 3.60 mmol). The reaction mixture was stirred at room temprature for 8 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-083 as a white solid (40 mg, yield: 15%). LC-MS 377 (M+H) ⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.76 (s, 1 H), 8.51 (s, 1 H), 8.32 (t, J = 8.4 Hz, 2 H), 7.76 (d, J = 7.6 Hz, 1 H), 7.67 (d J = 7.6 Hz, 1 H), 7.57 (s, 1 H), 7.49 (t, J = 7.2 Hz, 1 H), 7.32 (d, J = 8.0 Hz, 1 H), 7.16 (d, J = 8.0 Hz, 1 H), 5.05-4.99 (m, 1 H), 4.75 (s, 1 H), 3.37-3.30 (m, 2 H), 3.06-2.96 (m, 2 H), 2.40 (s, 2 H), 1.22 (s, 6 H).

Example 117: Preparation of compound A29-084

To a solution of tert-butyl 3-oxopyrrolidine-l-carboxylate (740 mg, 4.0 mmol) in EtOH (15 mL) was added 2-(ethylamino) ethanol (356 mg, 4.0 mmol) at room temperature with stirring. After stirring continued for 0.5 h at room temperature, NaBH₃CN (377 mg, 6.0 mmol) was added at 0 °C. The resulting mixture was stirred at room temperature overnight. The mixture was quenched slowly with H₂0 (10 ml), then evaporated and extracted with EtOAc (3 >< 30 mL). The organic layer was dried over Na₂S0₄, filtered, and concentrated. The residue of compound SP-0011265-103 (500 mg, crude) as a brown oil was used in next step of reaction.

A mixture of SP-0011265-103 (200 mg, crude) and dioxane-HCl (1.5 mL) in DCM (5 mL) was stirred at room temperature for 2 h. DCM was removed by rotary evaporation. The crude compound SP-0011265-114 was obtained as a brown solid used in next step of reaction without further purification.

A mixture of 4-chloro-6-fluoroquinazoline (150 mg, 0.82 mmol), 2- (ethyl(pyrrolidin-3-yl)amino)ethanol (150 mg, crude) and triethylamine (0.5 mL, 3.5 mmol) in isopropanol (5 mL) was stirred at 70 °C overnight. The mixture was cooled down and isopropanol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-084 as a brown solid (56 mg, yield: 29%). LC-MS 305 (M+H)⁺, purity 95% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.44 (s, 1 H), 7.97 (dd, J = 10.8 Hz, 2.8 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.69 (td, J = 9.2 Hz, 2.8 Hz, 1 H), 4.42-4.43 (m, 1 H), 3.86-4.01 (m, 3 H), 3.65 (t, J = 10.4 Hz, 1 H), 3.34-3.41 (m, 3 H), 2.58-2.69 (m, 4 H), 2.13-2.16 (m, 1 H), 1.80-1.85 (m, 1 H), 0.99 (t, J = 7.2 Hz, 3 H). Example 118: Preparation of compound A29-085

A29-085

To a solution of SP-0011321-041 (280 mg, 0.870 mmol) and triethylamine (176 g, 1.74 mmol) in THF (10 mL) was added Boc₂0 (284 mg, 1.30 mmol). The resulting solution was heated to 60 °C for 2 h. LC-MS analysis showed completed consumption of SP- 0011321-041. The mixture was cooled down and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (using petroleum ether: EtOAc = 2: 1 - 1 :2) to afford the compound SP-0011321-044 (310 mg, yield: 85%). LC-MS 423 (M+H)⁺, purity 71% (UV 214 nm).

To a solution of compound SP-0011321-044 (120 mg, 0.284 mmol) in 1,4-dioxane (5 mL) was added a solution of Oxone (262 mg, 0.427 mmol) in water (3 mL). The reaction mixture was stirred at room temperature for 0.5 h and concentrated under reduced pressure. The residue was partitioned between EtOAc (20 mL) and brine (15 mL). The organic layer was separated and the aqueous phase was extracted with EtOAc (20 mL). The combined organic phase was dried over Na₂S0₄, filtered and concentrated to give a crude product SP- 0011321-051 (90 mg, yield: 70%>) which was used in the next step of reaction without further purification.

A mixture of SP-0011321-051 (90 mg, 0.198 mmol) and trifiuoracetic acid (3 mL) in DCM (5 mL) was stirred at room temperature for 2 h, and concentrated under reduced pressure. The residue was partitioned between EtOAc (20 mL) and saturated Na₂C0₃ (15 mL). The organic layer was separated and the aqueous phase was extracted with EtOAc (15 mL). The combined organic phase was combined, dried over Na₂S0₄, filtered and concentrated. The residue was purified by Pre-HPLC to give the product A29-085 as a white solid (20 mg, yield: 20%). LC-MS 355 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.04 (d, J = 2.0 Hz, 1 H), 8.90 (d, J = 6.8 Hz, 1 H), 8.62 (s, 1 H), 8.18 (dd, J = 8.8 Hz, 2.0 Hz, 1 H), 7.86 (d, J = 8.8 Hz, 1 H), 6.89 (d, J = 8.0 Hz, 1 H), 6.48 (s, 1 H), 6.40 (dd, J = 8.0 Hz, 2.0 Hz, 1 H), 5.03-4.97 (m, 1 H), 4.86 (s, 2 H), 3.28 (s, 3 H), 3.26-3.18 (m, 2 H), 2.92 (td, J = 15.6 Hz, 6.4 Hz, 2 H).

Example 119: Preparation of compound A29-086

A02-171 SP-0011321 -057 A29-086

To a solution of compound A02-171 (200 mg, 0.608 mol) in DMF (4 mL) was added 2-(tert-butoxycarbonylamino)acetic acid (143 mg, 0.816 mmol), HATU (310 mg, 0.816 mmol) and triethylamine (678 mg, 6.08 mmol). The mixture was stirred at 40 °C for 5 h. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (2 x 30 mL). The organic layer was washed with water (15 mL), brine (15 mL), dried over Na₂S0₄, filtered, and concentrated under reduced pressure to afford crude product SP-0011321-057 (180 g, yield: 66%). The resulting crude product was used in the next step of reaction without further purification.

To a solution of compound SP-0011321-057 (180 mg, 0.400 mol) in DCM (5 mL) was added trifluoroacetic acid (3 mL). The reaction mixture was stirred at room temperature for 2 h and concentrated under reduced pressure. The residue was partitioned between EtOAc (30 mL) and saturated Na₂C0₃ (20 mL). The organic layer was separated and the aqueous phase was extracted with EtOAc (20 mL). The combined organic phase was dried over Na₂S0₄, filtered and concentrated. The residue was purified by Pre-HPLC to give the product A29-086 as a white solid (90 mg, yield: 64%). LC-MS 352 (M+H)⁺, purity 98% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.69 (s, 1 H), 8.51 (s, 1 H), 8.27 (d, J = 6.8 Hz, 1 H), 8.23 (dd, J = 10.0 Hz, 2.4 Hz, 1 H), 7.77 (dd, J = 9.2 Hz, 5.6 Hz, 1 H), 7.67 (td, J = 8.4 Hz, 2.8 Hz, 1 H), 7.60 (s, 1 H), 7.38 (d, J = 8.0 Hz, 1 H), 7.18 (d, J = 8.0 Hz, 1 H), 5.03-4.98 (m, 1 H), 3.39-3.29 (m, 2 H), 3.25 (s, 2 H), 3.00 (td, J = 16.0 Hz, 6.0 Hz, 2 H). Example 120: Preparation of compound A29-087

A29-087

To a solution of compound A02-171 (200 mg, 0.608 mol) in DMF (4 mL) was added 5-(2-(tert-butoxycarbonylamino)ethylamino)-5-oxopentanoic acid (224 mg, 0.816 mmol), HATU (310 mg, 0.816 mmol) and triethylamine (678 mg, 6.08 mmol). The mixture was stirred at 40 °C for 16 h. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (2 x 30 mL). The organic layer was washed with water (15 mL) and brine (15 mL). The combined organic layer was dried over Na₂S0₄, filtered, and concentrated to afford crude product SP-0011321-060 (180 g, yield: 48%). The resulting crude product was used in the next step of reaction without further purification.

To a solution of compound SP-0011321-060 (180 mg, 0.327 mol) in DCM (5 mL) was added trifluoroacetic acid (3 mL). The reaction mixture was stirred at room temperature for 2 h and concentrated under reduced pressure. The residue was partitioned between EtOAc (30 mL) and saturated Na₂C0₃ (20 mL). The organic layer was separated and the aqueous phase was extracted with EtOAc (20 mL). The combined organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give the product A29-087 as a white solid (110 mg, yield: 75%). LC-MS 451 (M+H)⁺, purity 98% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.87 (s, 1 H), 8.51 (s, 1 H), 8.26 (d, J = 6.8 Hz, 1 H), 8.22 (dd, J = 10.0 Hz, 2.8 Hz, 1 H), 7.81-7.75 (m, 2 H), 7.67 (td, J = 8.4 Hz, 2.8 Hz, 1 H), 7.58 (s, 1 H), 7.33 (d, J = 8.4 Hz, 1 H), 7.16 (d, J = 8.0 Hz, 1 H), 5.02-4.98 (m, 1 H), 3.35-3.29 (m, 2 H), 3.08-2.94 (m, 5 H), 2.56 (t, J = 6.4 Hz, 1 H), 2.29 (t, J = 7.2 Hz, 2 H), 2.14-2.11 (m, 2 H), 1.84-1.78 (m, 2 H). Example 121: Preparation of compound A29-088

To a solution of compound A23-001 (250 mg, 0.83 mmol) in DMF (5.0 mL) were added 2-(tert-butoxycarbonylamino)acetic acid ( 290 mg, 1.66 mmoL), HATU (946 mg, 2.49 mmol) and triethylamine (0.60 mL, 4.15 mmol). The reaction mixture was stirred at 50 °C for 8 h. The reaction mixture was cooled down, extracted with ethyl ether (200 mL) and washed with water (50 mL).The organic extracts were dried over Na₂S0₄, filtered and concentrated to yield a crude product SP-0011507-076 (274 mg, yield: 72%) which was used for next step of reaction without further purification.

To a solution of compound SP-0011507-076 (274 mg, 0.60 mmol) in DCM (6.0 mL) was added trifluoro acetic acid (3.0 mL) at 0 °C. The mixture was stirred at room temperature for 1 h. DCM was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-088 as a white solid (82 mg, yield: 38%). LC-MS 359 (M+H)⁺, purity 98% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.98 (s, 1 H), 8.67 (d, J = 6.4 Hz, 1 H), 8.62 (s, 1 H), 8.07 (d, J = 8.8 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.62 (s, 1 H), 7.39 (d, J = 8.0 Hz, 1 H), 7.19 (d, J = 8.0 Hz, 1 H), 5.04-4.99 (m, 1 H), 3.40-3.36 (m, 2 H), 3.25 (s, 2 H), 3.06-2.96 (m, 2 H).

Example 122: Preparation of compound A29-089

To a solution of compound A23-001 (250 mg, 0.83 mmol) in DMF (5.0 mL) were added 5-(2-(tert-butoxycarbonylamino)ethylamino)-5-oxopentanoic acid (455 mg, 1.66 mmoL), HATU (946 mg, 2.49 mmol) and triethylamine ( 0.60 mL, 4.15 mmol). The reaction mixture was stirred at 50 °C for 8 h. The reaction mixture was cooled down, extracted with ethyl ether (200 mL), washed with water (50 mL). The organic extracts were dried over Na₂S0₄, filtered and concentrated under reduced pressure to yield a crude product SP-0011507-075 (324 mg, yield: 70%) which was used in the next step of reaction without further purification.

To a solution of compound SP-0011507-075 (324 mg, 0.58 mmol) in DCM (6.0 mL) was added trifluoro acetic acid (3.0 mL) at 0 °C. The mixture was stirred at room temperature for 1 h. DCM was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-089 as a white solid (104 mg, yield: 39%). LC-MS 458 (M+H)⁺, purity 95% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.85 (d, J = 14.4 Hz, 1 H), 8.97 (s, 1 H), 8.67 (d, J = 6.8 Hz, 1 H), 8.62 (s, 1 H), 8.07 (d, J = 8.8 Hz, 1 H), 7.88 (t, J = 5.2 Hz, 1 H), 7.79 (d, J = 8.8 Hz, 1 H), 7.59 (s, 1 H), 7.34 (d, J = 8.4 Hz, 1 H), 7.17 (d, J = 8.0 Hz, 1 H), 5.04-4.99 (m, 1 H), 3.40-3.36 (m, 2 H), 3.32-3.30 (m, 2 H), 3.07-3.03 (m, 2 H), 3.01-2.95 (m, 2 H), 2.58-2.55 (m, 2 H), 2.29 (t, J = 7.2 Hz, 2 H), 2.15-2.09 (m, 2 H), 1.84-1.76 (m, 2 H).

Example 123: Preparation of compound A29-090 and A29-091

To a cold solution of K O3 (253 mg, 2.50 mmol) in H₂S0₄ (5 mL) was added slowly 7-chloroquinazolin-4-olhydrochloride (500 mg, 2.78 mmol) at 0 °C. The reaction mixture was stirred at 100 °C for 5 h. The mixtures was poured into a mixture of ice and water and a precipitate was formed. The precipitate was isolated, washed with water and dried under vacuum to give a crude product SP-0011507-053 (470 mg, yield: 84%>) which was used in the next step of reaction without further purification.

A mixture of SP-0011507-053 (470 mg, 2.09 mmol), thionyl dichloride (7.40 g, 62.7 mol) and catalytic amount of anhydrous DMF (0.50 mL) was heated to reflux for 24 h. The mixture was cooled down and excess thionyl dichloride was removed by rotary evaporation. DCM (10 mL) was added to dissolve the solid completely, and addition of petroleum ether resulted in a precipitate. The precipitate was filtered off and washed with petroleum ether to provide crude product SP-0011507-064 (300 mg, yield: 59%) which was used in the next step of reaction without further purification.

A mixture of SP-0011507-064 (300 mg, 1.23 mmol), 2,3-dihydro-lH-indene-2,5- diamine (200 mg, 1.35 mmol) and triethylamine (0.53 mL, 3.69 mmol) in isopropyl alcohol (30 mL) was heated to 50 °C for 8 h. The mixture was cooled down and excess of isopropyl alcohol was removed by rotary evaporation. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29-090 (140 mg) and A29-091 (35 mg).

A29-090 LC-MS 356 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.30 (s, 1 H), 8.95 (d, J = 6.4 Hz, 1 H), 8.63 (s, 1 H ), 7.98 (s, 1 H), 6.89 (d, J = 8.0 Hz, 1 H), 6.48 (s, 1 H), 6.40 (d J = 7.6 Hz, 1 H), 4.98-4.92 (m, 1 H), 4.87 (s, 2 H), 3.33-3.18 (m, 2 H), 2.93-2.84 (m, 2 H).

A29-091 LC-MS 356 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 8.87 (s, 1 H), 8.63 (d, J = 8.4 Hz, 1 H), 8.57 (s, 1 H), 7.83 (d, J = 8.4 Hz, 1 H), 6.89 (d J = 6.0 Hz,l H), 6.45 (s, 1 H), 6.40 (d, J = 7.6 Hz, 1 H), 4.98-4.92 (m, 1 H), 4.87 (s, 2 H), 3.25-3.20 (m, 2 H), 2.94-2.85 (m, 2 H).

Example 124: Preparation of compound A29-093

To a solution of compound A29-086 (100 mg, 0.280 mol) in DMF (4 mL) were added biotin (82 mg, 0.336 mmol), HATU (127 mg, 0.336 mmol) and triethylamine (283 mg, 2.80 mmol). The mixture was stirred at 40 °C for 16 h. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (2 x 30 mL). The organic layer was washed with water (15 mL) and brine (15 mL), dried over Na₂S0₄, filtered and concentrated under reduced pressure. The residue was purified by Prep-HPLC to give A29- 093 as a white solid (60 mg, yield: 37%). LC-MS 578 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.88 (s, 1 H), 8.53 (s, 1 H), 8.27 (d, J = 6.4 Hz, 1 H), 8.23 (dd, J = 10.0 Hz, 2.4 Hz, 1 H), 8.14 (t, J = 6.0 Hz, 1 H), 7.77 (dd, J = 9.2 Hz, 6.0 Hz, 1 H), 7.67 (td, J = 8.4 Hz, 2.8 Hz, 1 H), 7.55 (s, 1 H), 7.34 (d, J = 8.0 Hz, 1 H), 7.18 (d, J = 8.0 Hz, 1 H), 6.43 (s, 1 H), 6.37 (s, 1 H), 5.03-4.97 (m, 1 H), 4.33-4.29 (m, 1 H), 4.16-4.12 (m, 1 H), 3.85 (d, J = 6.8 Hz, 2 H), 3.37-3.29 (m, 2 H), 3.13-3.08 (m, 1 H), 3.00 (td, J = 15.6 Hz, 5.6 Hz, 2 H), 2.82 (dd, J = 12.8 Hz, 5.2 Hz, 1 H), 2.57 (d, J = 12.4 Hz, 1 H), 2.17 (t, J = 7.2 Hz, 2 H), 1.64-1.31 (m, 6 H).

Example 125: Preparation of compound A29-094

A29-087 A29-094

To a solution of compound A29-087 (50 mg, 0.111 mol) in DMF (3 mL) were added biotin (32 mg, 0.133 mmol), HATU (51 mg, 0.133 mmol) and triethylamine (112 mg, 1.11 mmol). The mixture was stirred at 40 °C for 16 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (2 x 20 mL). The organic layer was washed with water (10 mL), brine (10 mL), dried over Na₂S0₄, filtered, and concentrated under reduced pressure. The residue was purified by Prep-HPLC to give A29-094 as a white solid (30 mg, yield: 40%). LC-MS 677 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.82 (s, 1 H), 8.52 (s, 1 H), 8.27 (d, J = 6.4 Hz, 1 H), 8.23 (dd, J = 10.0 Hz, 2.8 Hz, 1 H), 7.85-7.82 (m, 2 H), 7.77 (dd, J = 9.2 Hz, 5.6 Hz, 1 H), 7.67 (td, J = 9.2 Hz, 2.8 Hz, 1 H), 7.58 (s, 1 H), 7.33 (d, J = 8.4 Hz, 1 H), 7.16 (d, J = 8.4 Hz, 1 H), 6.43 (s, 1 H), 6.37 (s, 1 H), 5.02-4.97 (m, 1 H), 4.31-4.29 (m, 1 H), 4.14-4.10 (m, 1 H), 3.37-3.29 (m, 2 H), 3.11-2.98 (m, 7 H), 2.81 (dd, J = 12.4 Hz, 4.8 Hz, 1 H), 2.57 (d, J = 12.4 Hz, 1 H), 2.29 (t, J = 7.6 Hz, 2 H), 2.10 (t, J = 7.6 Hz, 2 H), 2.05 (t, J = 7.6 Hz, 2 H), 1.84-1.76 (m, 2 H), 1.63-1.27 (m, 6 H). Example 126: Preparation of Compound A29-095

To a solution of compound SP-0011507-078 (128 mg, 0.35 mmol) in DMF (2.0 mL) were added biotin (171 mg, 0.70 mmoL), HATU (334 mg, 0.88 mmol) and triethylamine (0.25 mL, 1.75 mmol). The reaction mixture was stirred at 50 °C for 8 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29- 095 as a white solid (50 mg, yield: 25%). LC-MS 585 (M+H) ⁺, purity 97% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.89 (s, 1 H), 8.98 (s, 1 H), 8.68 (d, J = 6.4 Hz, 1 H), 8.62 (s, 1 H), 8.14 (t, J = 6.0 Hz, 1 H), 8.07 (d, J = 8.4 Hz, 1 H), 7.79 (d, J = 8.4 Hz, 1 H), 7.56 (s, 1 H), ), 7.34 (d, J = 8.4 Hz, 1 H), 7.19 (d, J = 8.0 Hz, 1 H), 6.43 (s, 1 H), 6.37 (s, 1 H), 5.04-5.00 (m, 1 H), 4.32-4.28 (m, 1 H), 4.15-4.12 (m, 1 H), 3.85 (d, J = 5.6 Hz, 1 H), 3.39-3.35 (m, 2 H), 3.12-3.08 (m, 1 H), 3.05-2.96 (m, 2 H), 2.84-2.80 (m, 1 H), 2.59-2.56 (m, 1 H), 2.16 (t, J = 6.8 Hz, 2 H), 1.62-1.30 (m, 6 H).

Example 127: Preparation of compound A29-096

To a solution of compound SP-0011507-079 (133 mg, 0.29 mmol) in DMF (2.0 mL) were added biotin ( 106 mg, 0.44 mmoL), HATU (330 mg, 0.87 mmol) and triethylamine (0.21 mL, 1.45 mmol). The reaction mixture was stirred at 50 °C for 8 h. The resulting mixture was evaporated and the residue was purified by Prep-HPLC to give A29- 096 as a white solid (20 mg, yield: 10%). LC-MS 684 (M+H) ⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.82 (s, 1 H), 8.98 (s, 1 H), 8.67 (d, J = 6.0 Hz, 1 H), 8.62 (s, 1 H), 8.07 (d, J = 8.4 Hz, 1 H), 7.85 (s, 1 H), 7.79 (d, J = 8.8 Hz, 2 H), 7.58 (s, 1 H), 7.34 (d, J = 5.2 Hz, 1 H), 7.17 (d, J = 8.8 Hz, 1 H), 6.42 (s, 1 H), 6.36 (s, 1 H), 5.04-4.95 (m, 1 H), 4.32-4.25 (m, 1 H), 4.15-4.08 (m, 1 H), 3.11-3.05 (m, 6 H), 3.00 (d, J = 5.6 Hz, 2 H), 2.89-2.78 (m, 2 H), 2.33-2.32 (m, 1 H), 2.28 (t, J = 8.0 Hz, 2 H), 2.11 (t, J = 7.6 Hz, 2 H), 2.05 (t, J = 8.0 Hz, 2 H), 1.82-1.77 (m, 2 H), 1.62-1.20 (m, 6 H).

Example 128: Preparation of compound A29-097

To a solution of A29-090 (100 mg, 0.28 mmol) in DCM (20 mL) were added acetic anhydride (57 mg, 0.56 mmoL), 4-dimethylamiopryidine (3.6 mg, 0.03 mmoL) and triethylamine (0.12 mL, 0.84 mol). The mixture was stirred at room temperature for 6 h. DCM was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A29-097 as a white solid (35 mg, yield: 31%). LC-MS 398 (M+H)⁺, purity 97% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.86 (s, 1 H), 9.28 (s, 1 H), 8.98 (d, J = 6.4 Hz, 1 H), 8.65 (s, 1 H), 8.32 (s, 1 H), 7.99 (s, 1 H), 7.57 (s, 1 H), 7.32 (d, J = 8.0 Hz, 1 H), 7.17 (d, J = 8.0 Hz, 1 H), 5.04-4.99 (m, 1 H), 3.39-3.26 (m, 2 H), 3.05-2.96 (m, 2 H), 2.02 (s, 3 H).

Example 129: Preparation of compound A30-008

A30-008

To a solution of 2-hydroxybenzaldehyde (5.0 g, 40.984 mmol) in acrylonitrile (10.86 g, 0.204 mol) was added DABCO (942 mg, 8.2 mmol). The mixture was stirred at 90 °C for 20 h under N₂ atmosphere. The mixture was cooled down and the solvent was removed by rotary evaporation. The crude product was extracted with DCM (2 x 80 mL). The organic layer was washed successively with water (2 x 30 mL), brine (2 x 20 mL), dried over Na₂S0₄, filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography (using petroleum ether/EtOAc = 10:0— 7:3) to give compound SP-0010418-145-2 (5.1 g, yield: 80%). 1H NMR (400 MHz, CDC13): δ 730-7.26 (m, 1 H), 7.13-7.11 (m, 1 H), 6.99-6.97 (m, 1 H), 6.87-6.84 (m, 1 H), 4.82 (d, J = 1.2 Hz, 1 H).

A mixture of SP-0010418-145-2 (5.1 g, 32.484 mmol) in aqueous NaOH (10%,100 mL) was heated at 100 °C for 6 h. The reaction mixture was cooled down. Aqueous HC1 (5 N) solution was added dropwise to adjust the pH value of the reaction mixture to about 3 to result in the formation of a precipitate. The precipitate was filtered, dried under vacuum to give compound SP-0010418-145-3 (2.8 g, yield: 49%). LC-MS 177 (M+H)⁺, purity 96% (UV 214 nm);

A solution of DPP A (4.812 g, 17.494 mmol) in toluene (24 mL) was added dropwise at room temperature to a solution of compound SP-0010418-145-3 (2.8 g, 15.9 mmol) in DCM (80 mL) and triethylamine (4.0 mL). The mixture was stirred at 50 °C for 1 h, and more toluene (50 mL) was added. The resulting reaction mixture was heated at 85 °C for 3 h. The reaction mixture was cooled down to room temperature, aqueous HC1 (6.0 N, 28 mL) was added and the resulting reaction mixture was heated at reflux for an additional 3 h. The organic layer was separated, washed with saturated NaHC0₃, dried over Na₂S0₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (using petroleum ether/EtOAc = 10:0 - 7:3) to give compound SP-0010418-145-4 (2.0 g, yield: 85%). 1H NMR (400 MHz, CDC1₃): δ 7.55- 7.51 (m, 1 H), 7.24-7.22 (m, 1 H), 87.06-7.03 (m, 1 H), 7.4.42 (s, 2 H), 3.61 (s, 2 H).

A mixture of compound SP-0010418-145-4 (2.0 g, 13.513 mmol), 10% Pd/C (200 mg) and HC0₂NH₄ (8.0g) in MeOH (50 mL) was stirred at 60 °C for 18 h. The reaction mixture was filtered and concentrated under reduced pressure. The residue was diluted with saturated NaHC0₃ (20 mL) and extracted with EtOAc (50 mL). The organic layer was dried over Na₂S0₄, filtered and concentrated to give a crude SP-0010418-145-5 (1.1 g, yield: 55%) which was used in the next step of reaction without further purification. LC- MS 150 (M+H)⁺. To a solution of 4-chloro-6-fluoroquinazoline (610 mg, 3.355 mmol) in isopropanol (10 mL) were added SP-0010418-145-5 (500 mg, 3.355 mmol) and triethylamine (1.5 mL). The mixture was stirred at 70 °C for 8 h. The mixture was cooled down and isopropanol was removed by rotary evaporation. The residue was purified by Prep-HPLC to give the title compound A30-008 (20 mg, two steps yield: 2 %). LC-MS 295 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6): δ 8.53 (s, 1 H), 8.26-8.23 (m, 1 H), 8.05 (d, J = 6.8 Hz, 1 H), 7.82-7.68 (m, 2 H), 7.16-7.10 (m, 2 H), 6.92-6.82 (m, 2 H), 4.75-4.66 (m, 1 H), 4.37-4.33 (m, 1 H), 4.00-3.95 (m, 1 H), 3.20-3.02 (m, 2 H).

Example 130: Preparation of compound A30-009

A02-186 A30-009

To a solution of compound A02-186 (50 mg, 0.162 mmol) in DMF (3 mL) were added 2-hydroxyacetic acid (19 mg, 0.244 mmol), HATU (123 mg, 0.324 mmol) and triethylamine (82 mg, 0.810 mmol). The mixture was stirred at 40 °C for 16 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (2 x 15 mL). The organic layer was washed with water (10 mL) and brine (10 mL), dried over Na₂S0₄, filtered, and concentrated to give a residue. The residue was purified by Prep-HPLC to give A30-009 as a white solid (40 mg, yield: 67%). LC-MS 367 (M+H)⁺, purity 100% (UV 214 nm), 1H NMR (400 MHz, DMSO-d6) δ 9.52 (s, 1 H), 8.48 (s, 1 H), 8.25 (dd, J = 10.0 Hz, 2.8 Hz, 1 H), 8.06 (d, J = 7.2 Hz, 1 H), 7.77 (dd, J = 9.2 Hz, 5.6 Hz, 1 H), 7.69 (td, J = 8.4 Hz, 2.8 Hz, 1 H), 7.51 (s, 1 H), 7.42 (dd, J = 8.4 Hz, 2.0 Hz, 1 H), 7.04 (d, J = 8.4 Hz, 1 H), 5.65 (t, J = 5.6 Hz, 1 H), 4.56-4.52 (m, 1 H), 3.98 (d, J = 5.2 Hz, 2 H), 3.13 (dd, J = 16.0 Hz, 5.2 Hz, 1 H), 2.89-2.79 (m, 3 H), 2.18-2.14 (m, 1 H), 1.87-1.81 (m, 1 H).

Example 131: Preparation of compound A30-012

To a solution of compound A02-186 (50 mg, 0.162 mmol) in DMF (3 mL) were added 3-hydroxy-3-methylbutanoic acid (57 mg, 0.487 mmol), HATU (185 mg, 0.487 mmol) and triethylamine (82 mg, 0.810 mmol). The mixture was stirred at 60 °C for 16 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (2 x 15 mL). The organic layer was washed with water (10 mL) and brine (10 mL), dried over Na₂S0₄, filtered, and concentrated. The residue was purified by Prep-HPLC to give A30- 012 as a white solid (20 mg, yield: 30%). LC-MS 409 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, MeOD-d4) δ 8.46 (s, 1 H), 8.05 (dd, J = 9.6 Hz, 2.8 Hz, 1 H), 7.78 (dd, J = 9.2 Hz, 5.2 Hz, 1 H), 7.63 (td, J = 8.0 Hz, 2.8 Hz, 1 H), 7.38 (s, 1 H), 7.30 (dd, J = 8.4 Hz, 2.4 Hz, 1 H), 7.08 (d, J = 8.4 Hz, 1 H), 4.68-4.60 (m, 1 H), 3.24 (dd, J = 15.6 Hz, 4.8 Hz, 1 H), 3.08-2.86 (m, 3 H), 2.53 (s, 2 H), 2.32-2.27 (m, 1 H), 1.98-1.88 (m, 1 H) , 1.35 (s, 6 H).

Example 132: Preparation of compound A30-010

A02-186 A30-010

Acetic anhydride (20 mg, 0.195 mmol) was added to a solution of compound A02- 186 (50 mg, 0.162 mmol), triethylamine (164 mg, 1.62 mmol) and DMAP (2 mg, 0.016 mmol) in DCM (5 mL) cooled in an ice-bath. The reaction mixture was stirred at room temperature for 2 h, and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give A30-010 (25 mg, yield: 43%). LC-MS 351 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, MeOD-d4) δ 8.46 (s, 1 H), 8.05 (dd, J = 10.0 Hz, 2.8 Hz, 1 H), 7.78 (dd, J = 8.8 Hz, 5.2 Hz, 1 H), 7.63 (td, J = 8.0 Hz, 2.4 Hz, 1 H), 7.36 (s, 1 H), 7.30 (dd, J = 8.0 Hz, 2.0 Hz, 1 H), 7.08 (d, J = 8.0 Hz, 1 H), 4.67-4.60 (m, 1 H), 3.23 (dd, J = 16.0 Hz, 4.8 Hz, 1 H), 3.04-2.86 (m, 3 H), 2.31-2.26 (m, 1 H), 2.13 (s, 3 H), 1.98-1.89 (m, 1 H). Example 133: Preparation of compound A30-011

A02-186 A30-011

Isobutyryl chloride (21 mg, 0.244 mmol) was added to a solution of compound A02-186 (50 mg, 0.162 mmol), triethylamine (164 mg, 1.62 mmol) in DCM (5 mL) cooled in an ice-bath. The reaction mixture was stirred at room temperature for 2 h, and water (0.1 mL) was added to quench the reaction. The mixture was stirred for another 10 min and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give SP-

0010485-124 (25 mg, yield: 43%). LC-MS 379 (M+H)⁺, purity 96% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6) δ 9.70 (s, 1 H), 8.48 (s, 1 H), 8.24 (dd, J = 10.0 Hz, 2.8 Hz, 1 H), 8.05 (d, J = 7.2 Hz, 1 H), 7.77 (dd, J = 9.2 Hz, 5.6 Hz, 1 H), 7.69 (td, J = 9.2 Hz, 2.8 Hz, 1 H), 7.43 (s, 1 H), 7.32 (dd, J = 8.4 Hz, 2.0 Hz, 1 H), 7.02 (d, J = 8.4 Hz, 1 H), 4.55-4.51 (m, 1 H), 3.11 (dd, J = 16.0 Hz, 4.8 Hz, 1 H), 2.90-2.78 (m, 3 H), 2.61-2.50 (m, 1 H), 2.17- 2.00 (m, 1 H), 1.86-1.81 (m, 1 H), 1.19 (d, J = 6.8 Hz, 6 H).

Example 134: Preparation of compound A31-009

SP-0010020-187 A31 -009

To a solution of 2-ethoxyacetic acid (500 mg, 4.81 mmol) in CH2CI2 (25 mL) was added SOCl₂ (2 ml) at 0 °C slowly. The mixture was stirred at 60 °C for 4 h. The resulting mixture was evaporated to give a residue, which was used in the next step of reaction without further purification. LC-MS 123 (M+H)⁺, purity 60% (UV 214 nm).

To a solution of compound A02-189 (100 mg, 0.32 mmol) in DCM (10 mL) were added compound 2-ethoxyacetyl chloride (58 mg, 0.48 mmol) and triethylamine (0.5 mL, 3.6 mmol). The mixture was stirred at room temperature for 20 h. DCM was removed by rotary evaporation. The residue was purified by Prep-HPLC to give A31-009 as a white solid (34 mg, yield: 26%), LC-MS 402 (M+H)⁺, purity 99% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 11.78 (s, 1 H), 8.49 (s, 1 H), 8.21-8.24 (m, 1 H), 8.09 (dd, J = 10.0 Hz, 2.4 Hz, 1 H), 7.66-7.79 (m, 2 H), 4.62-4.64 (m, 1 H), 4.14 (m, 2 H), 3.50-3.56 (m, 2 H), 3.14-3.19 (m, 1 H), 2.73-2.79 (m, 3 H), 2.15-2.17 (m, 1 H), 1.95-2.01 (m, 1 H), 1.15 (t, J = 6.8 Hz, 3 H).

Example 135: Preparation of compound A31-010

A02-189 A31 -010

To a solution of compound A02-189 (100 mg, 0.32 mmol) in DMF (3.0 mL) were added isopropyl hydrogen carbonate (66 mg, 0.64 mmol), HATU (290 mg, 0.77 mmol) and triethylamine (0.5 mL, 3.5 mmol). The mixture was stirred at 85 °C overnight. The resulting mixture was purified by Prep-HPLC to give A31-010 as a white solid (22 mg, yield: 16%). LC-MS 416 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 11.66 (s, 1 H), 8.49 (s, 1 H), 8.23 (dd, J = 9.6 Hz, 2 Hz, 1 H), 8.10 (d, J = 7.6 Hz, 1 H), 7.75-7.79 (m, 1 H),7.67-7.72 (m, 1 H), 4.61-4.63 (m, 1 H), 4.13 (s, 2 H), 3.65-3.68 (m, 1 H), 3.16 (dd, J = 15.2 Hz, 4 Hz, 1 H), 2.74-2.79 (s, 3 H), 2.15-2.18 (m, 1 H), 1.97-2.01 (m, 1 H), 1.13 (d, J = 6.0 Hz, 6 H).

Example 136: Preparation of compound A31-011

To a solution of compound A02-189 (150 mg, 0.48 mmol) in DMF (5.0 mL) were added phenyl 5-tert-butylisoxazol-3-ylcarbamate (250 mg, 0.96 mmol) and pyridine (0.15 mL, 1.44 mmol). The mixture was stirred at 80 °C overnight. The resulting mixture was purified by Prep-HPLC to give A31-011 as a white solid (94 mg, yield: 41%). LC-MS 481 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 10.33-10.36 (m, 1 H), 9.82-9.84 (m, 1 H), 8.49 (s, 1 H), 8.23 (dd, J = 10 Hz, 2.4 Hz, 1 H), 8.09 (d, J = 7.6 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.67-7.72 (m, 1 H), 6.54 (s, 1 H), 4.61-4.65 (m, 1 H), 3.08-3.16 (m, 1 H), 2.67-2.72 (m, 3 H), 2.13-2.17 (m, 1 H), 1.94-2.03 (m, 1 H), 1.29 (s, 9 H).

Example 137: Preparation of compound A31-012

A02-189 A31-012

Acetic anhydride (0.5 mL, 0.48 mmol) was added to a solution of A02-189 (100 mg, 0.32 mmol), triethylamine (0.5 mL, 3.5 mmol) and DMAP (10 mg, 0.08 mmol) in DCM (10 mL) cooled in an ice-bath. The reaction mixture was stirred at room temperature overnight, washed with aqueous HC1 (0.5 M, 10 mL) and water (10 mL). The organic layer was dried over Na₂S0₄, filtered, and concentrated. The residue was purified by Prep-HPLC to give A31-012 as a white solid (73 mg, yield: 64%). LC-MS 358 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.49 (s, 1 H), 8.23 (dd, J = 10.4 Hz, 1 H), 8.09 (d, J = 7.2 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.68 (td, J = 8.8 Hz, 2.8 Hz,l H), 4.61-4.63 (m, 1 H), 3.14 (dd, J = 16 Hz, 5.2 Hz, 1 H), 2.72-2.78 (s, 3 H), 2.13-2.17 (m, 4 H), 1.98-2.01 (m, 1 H).

Example 138: Preparation of compound A31-013

A02-189 SP-0011265-009 A31-013

To a solution of A02-189 (50 mg, 0.48 mmol) in DCM (10 mL) were added 2- chloroacetyl chloride (53 mg, 0.48 mmol) and triethylamine (0.5 mL, 3.5 mmol). The mixture was stirred at room temperature for 1 h. DCM was removed by rotary evaporation. The residue was washed with aqueous NaHC0₃ and dried underv acuum to give the product SP-0011265-009 (165 mg, crude) as a brown solid.

To a solution of compound SP-0011265-009 (165 mg, crude) in DMF (2 mL) were added lH-imidazole (25 mg, 0.38 mmol) and KI (5 mg, 0.03 mmol). The mixture was stirred at 120 °C for 1 h under microwave. The resulting mixture was purified by Prep- HPLC to give the title compound A31-013 (37 mg, yield: 34%) as a white solid. LC-MS 424 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.47 (s, 1 H), 8.23 (dd, J = 10.4 Hz, 2.8 Hz, 1 H), 8.05 (d, J = 7.6 Hz, 1 H), 7.73-7.77 (m, 1 H), 7.67 (td, J = 9.2 Hz, 2.8 Hz, 1 H), 7.55 (s, 1 H), 7.08 (s, 1 H), 6.82 (s, 1 H), 4.58 (s, 2 H), 2.99 (dd, J = 15.2 Hz, 4.8 Hz,l H), 2.63-2.70 (m, 3 H), 2.10-2.13 (m, 1 H), 1.88-1.925 (m, 1 H), 1.23 (s, 1H).

Exam le 139: Preparation of compound A31-014

A02-189 A31 -014

To a solution of compound A02-189 (100 mg, 0.32 mmol) in DMF (2.0 mL) cooled in an ice-bath were added 2,2,2-trichloro-l-(lH-pyrrol-2-yl)ethanone (134 mg, 0.64 mmol) and NaH (10 mg, 0.38 mmol). The mixture was stirred at room temperature overnight. The resulting mixture was purified by Prep-HPLC to give A31-014 as a white solid (34 mg, yield: 26%). LC-MS 409 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- d6) δ 11.84 (s, 1 H), 8.49 (s, 1 H), 8.25 (dd, J = 10 Hz, 2.8 Hz, 1 H), 8.19 (d, J = 7.2 Hz, 1 H), 7.76-7.79 (m, 1 H), 7.67-7.72 (m, 1 H), 7.34-7.35 (m, 1 H), 7.27(s, 1 H), 7.04 (s, 1 H), 6.18 (s, 1 H), 4.64-4.66 (m, 1 H), 3.17 (dd, J = 13.2 Hz, 2.4 Hz, 1 H), 2.75-2.80 (s, 3 H), 2.16-2.19 (m, 1 H), 2.00-2.01 (m, 1 H).

Example 140: Preparation of compound A31-015

SP-0011265-004 A31-015

A solution of 2-(chloromethyl)-6-nitro-lH-benzo[d]imidazole (2 g, crude) in ammonium hydroxide (50 mL) was stirred at room temperature overnight. The resulting mixture was evaporated to dryness to provide a crude product (800 mg) as an orange solid which was used in the next step of reaction without further purification.

To a solution of 4-chloro-6-fluoroquinazoline (500 mg, 2.7 mmol) in isopropanol (15 mL) were added compound (6-nitro-lH-benzo[d]imidazol-2-yl)methanamine (800 mg, crude) and triethylamine (1.3 mL, 10.0 mmol). The mixture was stirred at 75 °C overnight. The mixture was cooled down and isopropanol was removed by rotary evaporation. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 2: 1 - 1 :2) to give compound A31-015 (560 mg, yield: 60%) as an orange solid. 1H NMR (400 MHz, MeOH- 4): δ 12.91-12.92 (m, 1 H), 9.05 (t, J = 5.6 Hz, 1 H), 8.46 (s, 1 H), 8.39 (s, 1 H), 8.19 (dd, J = 10 Hz, 2.8 Hz, 1 H), 8.08 (dd, J = 8.8 Hz, 2.4 Hz, 1 H), 7.81-7.85 (m, 1 H), 7.73-7.78 (m, 1 H), 7.65 (d, J= 9.2 Hz, 1 H), 5.03 (d, J = 5.6 Hz, 2 H).

Example 141: Preparation of compound A31-016

SP-0011265-006 A31 -016

To a solution of A28-008 (1.1 g, 2.60 mmol) in THF (20 mL) were added Boc₂0 (1.1 g, 5.20 mmol) and triethylamine (0.5 mL, 3.5 mmol). The reaction mixture was stirred at 70 °C overnight. The mixture was cooled down and THF was removed by rotary evaporation. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 2: 1 - 1 :5) to give compound SP-0010020-131 (508 mg, yield: 37%) as a yellow solid. LC-MS 524 (M+H)⁺, purity 100% (UV 214 nm).

To a solution of compound SP-0010020-131 (250 mg, 0.48 mmol) in dry DMA (3 mL) was added CuCN (170 mg, 1.92 mmol). The mixture was stirred at 120 °C for 1 h under microwave. The resulting mixture was diluted by water (15 mL), extracted with EtOAc (3 x 30 mL) and washed with brine (20 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated to give a residue. The residue was purified by silica gel column chromatography (using MeOH : CH₂C1₂ = 0 - 10%>) to give compound SP- 0011265-006 (84 mg, yield: 41%) as a white solid. LC-MS 524 (M+H)⁺, purity 76.4% (UV 214 nm).

To a solution of compound SP-0011265-006 (84 mg, 0.15 mmol) in DCM (5 mL) was added dioxane-HCl (4 M,1.5 ml). The mixture was stirred at room temperature for 2 h. DCM was removed by rotary evaporation. The residue was purified by Prep-HPLC to give compound A31-016 as a white solid (11 mg, yield: 22%). LC-MS 323 (M+H) , purity 100% ( UV 214 nm); 1H NMR (400 MHz, MeOD- 4) δ 8.78 (d, J = 1.2 Hz, 1 H), 8.58 (s, 1 H), 8.02 (dd, J = 8.8 Hz, 1.6 Hz, 1 H), 7.82 (d, J = 8.8 Hz, 1 H), 4.70-4.77 (m, 1 H), 3.07- 3.12 (m, 1 H), 2.67-2.74 (m, 3 H), 2.22-2.25 (m, 1 H), 2.01-2.09 (m, 1 H).

Example 142: Preparation of compound A31-017

A02-189 A31 -017

To a solution of compound A02-189 (100 mg, 0.32 mmol) in DMF (3.0 mL) was added furan-2-carboxylic acid (42 mg, 0.38 mmol), HATU (180 mg, 0.47 mmol) and triethylamine (0.5 mL, 3.5 mmol). The mixture was stirred at 85 °C for overnight. The resulting mixture was purified by Prep-HPLC to give A31-017 as a white solid (41 mg, yield: 31%). LC-MS 410 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- d6) δ 12.46 (s, 1 H), 8.49 (s, 1 H), 8.24 (dd, J = 10 Hz, 2.8 Hz, 1 H), 8.11 (d, J = 7.6 Hz, 1 H), 7.99 (s, 1 H), 7.76-7.79 (m, 1 H),7.70 (td, J = 9.2 Hz, 2.8 Hz, 1 H), 7.60 (m, 1 H), 6.71- 6.73 (m, 1 H), 4.65-4.67 (m, 1 H), 3.19 (dd, J = 15.2 Hz, 5.2 Hz, 1 H), 2.75-2.81 (m, 3 H), 2.17-2.20 (m, 1 H), 2.00-2.04 (m, 1 H).

Example 143: Preparation of compound A31-018

A02-189 A31 -018

To a solution of compound A02-189 (100 mg, 0.32 mmol) in DMF (3.0 mL) were added 2-phenoxyacetic acid (57.8 mg, 0.38 mmol), HATU (180 mg, 0.47 mmol) and triethylamine (0.5 mL, 3.5 mmol). The mixture was stirred at 85 °C overnight. The resulting mixture was purified by Prep-HPLC to give A31-018 as a white solid (65 mg, yield: 45%>). LC-MS 450 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 12.20 (s, 1 H), 8.49 (s, 1 H), 8.23 (d, J = 9.2 Hz, 1 H), 8.10 (d, J = 7.62 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.67-7.71 (m, 1 H), 7.31 (t, J = 7.2 Hz, 2 H), 6.95-6.99 (m, 3 H), 4.83 (s, 2 H), 4.64- 4.65 (m, 1 H), 3.15-3.19 (m, 1 H), 2.74-2.77 (m, 3 H), 2.15-2.18 (m, 1 H), 1.97-2.01 (m, 1 H).

Example 144: Preparation of compound A31-019

A02-189

A31 -019

To a solution of compound A02-189 (100 mg, 0.32 mmol) in DMF (3.0 mL) were added methanesulfonyl chloride (55 mg, 0.48 mmol) and triethylamine (0.5 mL, 3.5 mmol). The mixture was stirred at 50 °C overnight. The resulting mixture was purified by Prep- HPLC to give A31-019 as a white solid (31 mg, yield: 28%). LC-MS 344 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 12.04 (s, 1 H), 8.49 (s, 1 H), 8.43 (s, 1 H), 8.23 (dd, J = 10 Hz, 2.8 Hz, 1 H), 8.09 (d, J = 7.6 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.68 (td, J = 9.2 Hz, 2.8 Hz,l H), 4.62-4.64 (m, 1 H), 3.16 (dd, J = 16 Hz, 5.2 Hz, 1 H), 2.74-2.80 (m, 3 H), 2.15-2.17 (m, 1 H), 1.95-2.01 (m, 1 H).

Exam le 145: Preparation of compound A31-020

A02-189 A31 -020

To a solution of compound A02-189 (100 mg, 0.32 mmol) in DMF (3.0 mL) were added methanesulfonyl chloride (55 mg, 0.48 mmol) and triethylamine (0.5 mL, 3.5 mmol). The mixture was stirred at 100 °C for overnight. The resulting mixture was purified by Prep-HPLC to give A31-020 as a white solid (20 mg, yield: 10%). LC-MS 394 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 12.34 (s, 1 H), 8.48 (s, 1 H), 8.21 (dd, J = 10.4 Hz, 2.8 Hz, 1 H), 8.05 (d, J = 7.2 Hz, 1 H), 7.74-7.79 (m, 1 H), 7.67-7.72 (m, 1 H), 4.60-4.62 (m, 1 H), 2.93 (dd, J = 15.6 Hz, 4.8 Hz, 1 H), 2.86 (s, 3 H), 2.58-2.60 (m, 3 H), 2.08-2.13 (m, 1 H), 1.92-1.99 (m, 1 H). Example 146: Preparation of compound A31-021

A02-189 A31 -021

To a solution of compound A02-189 (150 mg, 0.48 mmol) in DMF (4.0 mL) were added 1-hydroxycyclopropanecarboxylic acid (58 mg, 0.57 mmol), HATU (271 mg, 0.71 mmol) and triethylamine (0.5 mL, 3.5 mmol). The mixture was stirred at 85 °C overnight. The resulting mixture was purified by Prep-HPLC to give A31-021 as a white solid (39 mg, yield: 20%). LC-MS 400 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- d6) δ 11.18 (s, 1 H), 8.49 (s, 1 H), 8.23 (dd, J = 9.6 Hz, 2.4 Hz, 1 H), 8.10 (d, J = 7.2 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.69 (td, J = 8.8 Hz, 2.8 Hz, 1 H), 6.59 (s, 1 H), 4.63-4.66 (m, 1 H), 3.16 (dd, J = 15.2 Hz, 4.0 Hz, 1 H), 2.66-2.79 (m, 3 H), 2.15-2.18 (m, 1 H), 1.97-2.02 (m, 1 H), 1.18-1.24 (m, 2 H), 1.02-1.05 (m, 2 H).

Example 147: Preparation of compound A31-022

A02-189 A31 -022

To a solution of compound A02-189 (100 mg, 0.32 mmol) in DMF (3.0 mL) were added 2-(diethylamino)acetic acid (49 mg, 0.38 mmol), HATU (180 mg, 0.48 mmol) and triethylamine (0.5 mL, 3.5 mmol). The mixture was stirred at 85 °C overnight. The resulting mixture was purified by Prep-HPLC to give A31-022 as a white solid (34 mg, yield: 25%>). LC-MS 429 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 11.34 (s, 1 H), 8.49 (s, 1 H), 8.23 (dd, J = 10.4 Hz, 2.8 Hz, 1 H), 8.09 (d, J = 7.2 Hz, 1 H), 7.75- 7.79 (m, 1 H), 7.69 (td, J = 8.4 Hz, 2.8 Hz, 1 H), 4.62-4.63 (m, 1 H), 3.30 (s, 2 H), 3.13- 3.18 (m, 1 H), 2.73-2.79 (m, 3 H), 2.56-2.61 (m, 4 H), 2.14-2.17 (m, 1 H), 1.98-2.00 (m, 1 H), 0.99 (t, J = 7.2 Hz, 6 H). Example 148: Preparation of compound A31-023

A02-189 A31 -023

To a solution of compound A02-189 (150 mg, 0.48 mmol) in DMF (4.0 mL) were added 2-(ethylthio)acetic acid (69 mg, 0.57 mmol), HATU (271 mg, 0.71 mmol) and triethylamine (0.5 mL, 3.5 mmol). The mixture was stirred at 85 °C overnight. The resulting mixture was purified by Prep-HPLC to give A31-023 as a white solid (85 mg, yield: 42%). LC-MS 418 (M+H)⁺, purity 98% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 12.00 (s, 1 H), 8.49 (s, 1 H), 8.23 (dd, J = 10.0 Hz, 2.4 Hz, 1 H), 8.09 (d, J = 7.2 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.69 (td, J = 9.2 Hz, 2.8 Hz, 1 H), 4.60-4.65 (m, 1 H), 3.38 (s, 2 H), 3.16 (dd, J = 16.0 Hz, 6.0 Hz, 1 H), 2.71-2.79 (m, 3 H), 2.58-2.63 (m, 2 H), 2.14-2.17 (m, 1 H), 1.96- 2.03 (m, 1 H), 1.19 (t, J = 7.2 Hz, 3 H).

Example 149: Preparation of compound A31-024

A02-189 SP-0011265-059 A31-024

To a solution of compound A02-189 (400 mg, 1.26 mmol) in DMF (5.0 mL) were added 2-(ethylthio)acetic acid (228 mg, 1.9 mmol), HATU (718 mg, 1.89 mmol) and triethylamine (1.5 mL, 10.5 mmol). The mixture was stirred at 85 °C overnight. The resulting mixture was diluted with water (15 mL), extracted with EtOAc (3 x 50 mL) and washed with brine (20 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 2: 1 - 1 :5) to provide the compound SP-0011265-059 (330 mg, yield: 62%). LC-MS 418 (M+H)⁺, purity 97% (UV 214 nm).

To a solution of compound SP-0011265-059 (150 mg 0.36 mmol) in dioxane (8.0 mL) and H₂0 (2.0 ml) was added Oxone (331 mg, 615 mmol). The mixture was stirred at room temperature for 0.5 h, and 1,4-dioxane was removed by rotary evaporation. The residue was diluted by water (15 mL), extracted with EtOAc (3 x 50 mL) and washed with brine (20 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by Prep-HPLC to give A31- 024 as a white solid (30 mg, yield: 18%). LC-MS 450 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.49 (s, 1 H), 8.23 (dd, J = 10.0 Hz, 2.4 Hz, 1 H), 8.09 (d, J = 7.6 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.69-7.72 (m, 1 H), 4.64-4.65 (m, 1 H), 4.38 (s, 2 H), 3.28-3.30 (m, 2 H), 3.15-3.21 (m, 1 H), 2.75-2.79 (m, 3 H), 2.15-2.17 (m, 1 H), 1.99- 2.02 (m, 1 H), 1.27 (t, J = 7.2 Hz, 3 H).

Example 150: Preparation of compound A31-025

A02-189 A31-025

To a solution of compound A02-189 (300 mg, 0.95 mmol) in MeOH (10 mL) was added acetaldehyde (418 mg, 9.5 mmol) at room temperature under stirring. The stirring continued at room temperature for 2 h. The reaction mixture was cooled down to 0 °C and NaBH₃CN (88 mg, 1.5 mmol) was added at this temperature. The mixture was warmed up to room temperature and stirred at this temperature overnight. The reaction mixture was quenched slowly with H₂0 (10 mL), and MeOH was removed under reduced pressure. The remaining aqueous phase was extracted with EtOAc (3 x 50 mL). The combined organic phase was washed with brine (20 mL), dried over Na₂S0₄, filtered and evaporated to give a residue. The residue was purified by Prep-HPLC to give A31-025 as a white solid (180 mg, yield: 55%). LC-MS 344 (M+H)⁺, purity 96% (UV 214 nm); 1H NMR (400 MHz, CDC1₃- d3) δ 8.65 (s, 1 H), 7.84-7.87 (m, 1 H), 7.47-7.52 (m, 1 H), 7.29 (dd, J = 8.8 Hz, 2.8 Hz, 1 H), 5.62 (d, J = 7.6 Hz, 1 H), 4.93 (s, 1 H), 4.85-4.88 (m, 1 H), 3.17-3.31 (m, 3 H), 2.66- 2.79 (m, 3 H), 2.10-2.17 (m, 2 H), 1.28 (t, J = 7.2 Hz, 3 H).

Example 1 1: Preparation of compound A31-028

A mixture of 4-chloro-7-fluoroquinazoline (100 mg, 0.55 mmol), 4,5,6,7- tetrahydrobenzo[d]thiazole-2,6-diamine (180 mg, 0.55 mmol) and triethylamine (1.0 mL, 7.2 mmol) in isopropyl alcohol (5 mL) was heated to 70 °C under N₂ overnight. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation to give a residue. The residue was purified by Prep-HPLC to give A31-028 as a yellow solid (84 mg, yield: 48%). LC-MS 316 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, MeOD- 4) δ 8.47 (s, 1 H), 8.29-8.33 (m, 1 H), 7.32-7.37 (m, 2 H), 4.70- 4.72 (m, 1 H), 3.05-3.10 (m, 1 H), 2.66-2.72 (m, 3 H), 2.21-224 (m, 1 H), 2.03-2.06 (m, 1 H).

Example 152: Preparation of compound A31-026

A26-011 A31 -026

Acetic anhydride (0.5 mL, 0.48 mmol) was added to a solution of A26-011 (100 mg, 0.30 mmol), triethylamine (0.5 mL, 3.5 mmol) and DMAP (10 mg, 0.08 mmol) in DCM (10 mL) cooled in an ice-bath. The reaction mixture was stirred at room temperature overnight, and then washed with aqueous HC1 (0.5 M, 10 mL) and water (10 mL). The organic layer was dried over Na₂S0₄, filtered, and concentrated to give a residue. The residue was purified by Prep-HPLC to give A31-026 as a white solid (56 mg, yield: 50%). LC-MS 374 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 11.92 (s, 1 H), 8.52 (d, J = 72.4 Hz, 1 H), 8.50 (s, 1 H), 8.25 (d, J = 7.6 Hz, 1 H), 7.78-7.81 (m, 1 H), 7.71 (d, J = 8.8 Hz, 1 H), 4.61-4.64 (m, 1 H), 3.16-3.22 (m, 1 H), 2.72-2.79 (m, 3 H), 2.13-2.16 (m, 1 H), 2.11 (s, 3 H), 1.95-2.01 (m, 1 H).

Example 153: Preparation of compound A31-02

A31-027 A mixture of 4,7-dichloroquinazoline (400 mg, 2.0 mmol) and 4,5,6,7- tetrahydrobenzo[d]thiazole-2,6-diamine (662 mg, 2.0 mmol) and triethylamine (3.0 mL, 21.6 mmol) in isopropyl alcohol (15 mL) was stirred at 70 °C under N₂ overnight. The reaction mixture was cooled down and excess isopropyl alcohol was removed by rotary evaporation to give a residue. The residue was purified by silica gel column chromatography (using MeOH: CH₂C1₂ = 0 - 10%) to give compound A31-027 (603 mg, yield: 90%) as a yellow solid. LC-MS 332 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.49 (s, 1 H), 8.42 (d, J = 8.8 Hz, 1 H), 8.34 (d, J = 7.6 Hz, 1 H), 7.72 (d, J = 2.4 Hz, 1 H), 7.57 (dd, J = 8.8 Hz, 2.0 Hz, 1 H), 6.79 (s, 2 H), 4.54-4.62 (m, 1 H), 2.91-2.97 (m, 1 H), 2.61-2.67 (m, 3 H), 2.05-2.09 (m, 1 H), 1.87-1.97 (m, 1 H).

Example 154: Preparation of compound A31-030

Acetic anhydride (1.0 mL, 0.96 mmol) was added to a solution of A31-027(150 mg, 0.45 mmol), triethylamine (0.5 mL, 3.5 mmol) and DMAP (10 mg, 0.08 mmol) in DCM (10 mL) cooled in an ice-bath. The reaction mixture was stirred at room temperature overnight, and then washed with aqueous HC1 (0.5 M, 10 mL) and water (10 mL). The organic layer was dried over Na₂S0₄, filtered, and concentrated to give a residue. The residue was purified by Prep-HPLC to give A31-030 as a white solid (89 mg, yield: 53%). LC-MS 374 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 11.91 (s, 1 H), 8.50 (s, 1 H), 8.39 (d, J = 8.8 Hz, 1 H), 8.30 (d, J = 7.2 Hz, 1 H), 7.73 (d, J = 2.0 Hz, 1 H), 7.58 (dd, J = 8.8 Hz, 2.4 Hz,l H), 5.76 (s, 1 H), 4.61-4.63 (m, 1 H), 3.10-3.17 (m, 1 H), 2.73- 2.79 (m, 3 H), 2.15-2.16 (m, 1 H), 2.11 (s, 3 H), 1.97-2.00 (m, 1 H).

Example 155: Preparation of compound A31-029

A31 -028 A31 -029 Acetic anhydride (1.0 mL, 0.96 mmol) was added to a solution of A31-028 (150 mg, 0.48 mmol), triethylamine (0.5 mL, 3.5 mmol) and DMAP (10 mg, 0.08 mmol) in DCM (10 mL) cooled in an ice-bath. The reaction mixture was stirred at room temperature overnight, and then washed with aqueous HC1 (0.5 M, 10 mL) and water (10 mL). The organic layer was dried over Na₂S0₄ filtered, and concentrated to give a residue. The residue was purified by Prep-HPLC to give A31-029 as a white solid (89 mg, yield: 52%). LC-MS 358 (M+H)⁺, purity 98% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 1 1.92 (s, 1 H), 8.48 (s, 1 H), 8.43-8.47 (m, 1 H), 8.24 (d, J = 7.6 Hz, 1 H), 7.41-7.47 (m, 2 H), 4.61-4.64 (m, 1 H), 3.10-3.15 (m, 1 H), 2.73-2.79 (m, 3 H), 2.15-2.16 (m, 1 H), 2.11 (s, 3 H), 1.95-2.00 (m, 1 H).

Example 156: Preparation of compound A31-031

A02-189 A31-031

To a solution of A02-189 (150 mg, 0.48 mmol) in DMF (4.0 mL) were added biotin (139 mg, 0.57 mmol), HATU (271 mg, 0.71 mmol) and triethylamine (0.5 mL, 3.5 mmol). The mixture was stirred at 85 °C overnight. The resulting mixture was purified by Prep- HPLC to give A31-031 as a yellow solid (105 mg, yield: 40%). LC-MS 418 (M+H)⁺, purity 94% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 11.89 (s, 1 H), 8.48 (s, 1 H), 8.23 (dd, J = 10.4 Hz, 2.8 Hz, 1 H), 8.08 (d, J = 7.2 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.69-7.71 (m, 1 H), 6.44 (s, 1 H), 6.36 (s, 1 H), 4.61-4.64 (m, 1 H), 4.29-4.32 (m, 1 H), 4.12-4.15 (m, 1 H), 3.09-3.11 (m, 2 H), 2.80-2.84 (m, 1 H), 2.72-2.78 (m, 3 H), 2.56-2.59 (m, 1 H), 2.41 (t, J = 7.2 Hz, 2 H), 2.14-2.18 (m, 1 H), 1.95-1.98 (m, 1 H), 1.57-1.64 (m, 3 H), 1.47-1.50 (m, 1 H), 1.30-1.36 (m, 2 H). Example 157: Preparation of compound A31-032

SP-0011321-026 SP-0011321 -062 SP-0011321 -069

To a solution of SP-0011321-026 (300 mg, 1.43 mmol) and 4,5,6,7- tetrahydrobenzo[d]thiazole-2,6-diamine dihydrogen bromide (709 mg, 2.14 mmol) in isopropyl alcohol (30 mL) was added triethylamine (1.44 g, 14.3 mmol). The resulting solution was heated to 80 °C under N₂ for 16 h. The reaction mixture was cooled down and concentrated to give the crude compound SP-0011321-062.

To a solution of SP-0011321-062 and triethylamine (288 g, 2.86 mmol) in THF (30 mL) was added (Boc)₂0 (468 mg, 2.15 mmol). The resulting solution was heated to 60 °C for 2 h. The mixture was cooled down and concentrated. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 2: 1 - 1 :2) to afford SP- 0011321-069 (380 mg, yield: 60%). LC-MS 444 (M+H)⁺, purity 75% (UV 214 nm).

To a solution of compound SP-0011321-069 (200 mg, 0.451 mmol) in 1, 4-dioxane (6 mL) was added a solution of Oxone (416 mg, 0.677 mmol) in water (5 mL). The reaction mixture was stirred at room temperature for 0.5 h and concentrated under reduced pressure to give a residue. The residue was partitioned between EtOAc (30 mL) and brine (20 mL). The organic layer was separated and the aqueous phase was extracted with EtOAc (20 mL). The organic layer was dried over Na₂S0₄, filtered and concentrated to give a crude product SP-0011321-078 (139 mg, yield: 65%>) which was used in next step of reaction without further purification.

To a solution of compound SP-0011321-078 (139 mg, 0.293 mmol) in DCM (5 mL) was added trifluoroacetic acid (3 mL). The reaction mixture was stirred at room temperature for 2 h and concentrated. The residue was partitioned between EtOAc (20 mL) and saturated aquesous Na₂C0₃ (15 mL). The organic layer was separated and the aqueous phase was extracted with EtOAc (15 mL). The combined orgaic layer was dried over Na₂S0₄, filtered and concentrated to give a residue. The residue was purified by Pre-HPLC to give the product A31-032 as a white solid (20 mg, yield: 18%). LC-MS 376 (M+H)⁺, purity 98% (UV 214 nm); 1H NMR (400 MHz, MeOD-d4) δ 8.99 (d, J = 1.6 Hz, 1 H), 8.60 (s, 1 H), 8.27 (dd, J = 8.8 Hz, 2.0 Hz, 1 H), 7.90 (d, J = 8.8 Hz, 1 H), 4.81-4.75 (m, 1 H), 3.23 (s, 3 H), 3.11-3.07 (m, 1 H), 2.78-2.70 (m, 3 H), 2.27-2.22 (m, 1 H), 2.11-2.05 (m, 1 H).

Example 158: Preparation of compound A31-033

SP-0011321-078 SP-0011321-081 A31-033

To a solution of SP-0011321-078 (139 mg, 0.293 mmol) in DCM (5 mL) was added trifluoroacetic acid (3 mL). The reaction mixture was stirred at room temperature for 2 h and concentrated under reduced pressure to give a residue. The residue was partitioned between EtOAc (20 mL) and saturated aqueous Na₂C0₃ (15 mL). The organic layer was separated and the aqueous phase extracted with EtOAc (15 mL). The combined organic layer was dried over Na₂S0₄, filtered and concentrated to provide SP-0011321-081 (76 mg). The crude product was used in next step of reaction without further purification.

Acetic anhydride (41 mg, 0.406 mmol) was added to a solution of SP-0011321-081 (76 mg, 0.203 mmol), triethylamine (205 mg, 2.03 mmol) and DMAP (3 mg, 0.020 mmol) in DCM (6 mL) cooled in an ice-bath. The reaction mixture was stirred at room temperature for 2 h, and concentrated to give a residue. The residue was purified by Pre-HPLC to give the product A31-033 as a white solid (50 mg, yield: 59%). LC-MS 418 (M+H)⁺, purity 98% (UV 214 nm); 1H NMR (400 MHz, MeOD-d4) δ 11.93 (s, 1 H), 9.04 (d, J = 1.6 Hz, 1 H), 8.79 (d, J = 7.6 Hz, 1 H), 8.61 (s, 1 H), 8.21 (dd, J = 8.8 Hz, 2.0 Hz, 1 H), 7.88 (d, J = 8.8 Hz, 1 H), 4.68-4.66 (m, 1 H), 3.30 (s, 3 H), 3.15 (dd, J = 15.2 Hz, 5.2 Hz, 1 H), 2.84-2.77 (m, 3 H), 2.20-2.16 (m, 1 H), 2.12 (s, 3 H), 2.03-1.99 (m, 1 H).

Example 159: Preparation of compound A31-034

A02-189 SP-0011265-128 A31-034

To a solution of A02-189 (250 mg, 0.79 mmol) in DMF (4.0 mL) were added 2- (tert-butoxycarbonylamino)acetic acid (166 mg, 0.95 mmol), HATU (450 mg, 1.18 mmol) and triethylamine (1.5 mL, 10.5 mmol). The mixture was stirred at 85 °C overnight. The resulting mixture was diluted by water (15 mL), extracted with EtOAc (3 x 30 mL) and washed with brine (20 mL). The organic layer was dried over anhydrous Na₂S0₄, filtered and concentrated under reduced pressure to give the crude SP-0011265-128 (450 mg) as a brown soil, which was used in next step of reaction without further purification.

To a solution of SP-0011265-128 (450 mg, crude) in DCM (10 mL) was added dioxane-HCl (4 M, 3.5 mL). The reaction mixture was stirred at room temperature for 2 h. DCM was removed by rotary evaporation to give a residue. The residue was purified by Prep-HPLC to give compound A31-034 as a white solid (56 mg, yield: 18%). LC-MS 373 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.49 (s, 1 H), 8.23 (dd, J = 10.0 Hz, 2.8 Hz, 1 H), 8.10 (d, J = 7.6 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.67-7.72 (m, 1 H), 5.07-5.35 (m, 2 H), 4.61-4.64 (m, 1 H), 3.35 (s, 2 H), 3.15 (dd, J = 15.2 Hz, 5.2 Hz, 2 H), 2.72-2.79 (m, 3 H), 2.14-2.17 (m, 1 H), 1.96-2.01 (m, 1 H).

Example 160: Preparation of compound A31-035

To a solution of A02-189 (250 mg, 0.79 mmol) in DMF (4.0 mL) were added 5-(2- (tert-butoxycarbonylamino) ethylamino)-5-oxopentanoic acid (260 mg, 0.95 mmol), HATU (450 mg, 1.18 mmol) and triethylamine (1.5 mL, 10.5 mmol). The mixture was stirred at 85 °C overnight. The resulting mixture was diluted by water (15 mL), extracted with EtOAc (3 x 30 mL) and washed with brine (20 mL). The organic layer was dried over anhydrous Na₂S0₄, filtered and concentrated to give the crude SP-0011265-127 (500 mg) as brown soil, which was used in next step of reaction without further purification.

A mixture of SP-0011265-127 (500 mg, crude) and dioxane-HCl (3.5 ml) in DCM was stirred at room temperature for 2 h. DCM was removed by rotary evaporation to give a residue. The residue was purified by Prep-HPLC to give compound A31-035 as a yellow solid (67 mg, yield: 17%). LC-MS 472 (M+H)⁺, purity 100% ( UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.49 (s, 1 H), 8.23 (dd, J = 10.0 Hz, 2.8 Hz, 1 H), 8.09 (d, J = 7.6 Hz ,1 H), 7.75-7.79 (m, 2 H), 7.66-7.71 (m, 1 H), 4.61-4.63 (m, 1 H), 3.12-3.17 (m, 2 H), 3.01- 3.07 (m, 2 H), 2.72-2.78 (m, 3 H), 2.55 (t, J = 6.4 Hz, 2 H), 2.39 (t, J = 7.2 Hz, 2 H), 2.14- 2.17 (m, 1 H), 2.10 (t, J = 7.6 Hz, 2 H), 1.95-2.00 (m, 1 H), 1.76-1.83 (m,2 H). Example 161: Preparation of compound A31-036

A31-034 A31-036

A mixture of A31-034 (150 mg, 0.40 mmol), biotin (117 mg, 0.48 mmol), HATU (228 mg, 0.60 mmol) and triethylamine (0.5 mL, 3.5 mmol) in DMF (3.0 mL) was stirred at 85 °C overnight. The mixture was concentrated under reduced pressure to give a residue. The residue was purified by Prep-HPLC to give A31-036 as a yellow solid (37 mg, yield: 15%). LC-MS 599 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO- 6) δ 8.49 (s, 1 H), 8.18-8.24 (m, 2 H), 8.10 (d, J = 7.6 Hz, 1 H), 7.75-7.79 (m, 1 H), 7.66-7.72 (m, 1 H), 6.42 (s, 1 H), 6.36 (s, 1 H), 6.61-6.65 (m, 1 H), 4.30 (t, J = 5.6 Hz, 1 H), 3.08-3.18 (m, 2 H), 2.72-2.84 (m, 4 H), 2.57 (d, J = 12.4 Hz, 1 H), 2.16 (t, J = 7.2 Hz, 3 H), 1.96-2.01 (m, 1 H), 1.59-1.63 (m, 1 H), 1.44-1.55 (m, 3 H), 1.30-1.37 (m, 2 H).

Example 162: Preparation of compound A32-001

SP-0011379-028-A SP-0011379-028-B

SP-0011379-028-A SP-0011379-037 A32-001

To a solution of 3-aminocyclohexanol (2.3 g, 20.0 mmol) in H₂0/l,4-dioxane (20 mL/20 mL) were added Na₂C0₃ (2.52 g, 30.0 mmol) and benzyl carbonochloridate (5.1 g, 30.0 mmol). The mixture was stirred at room temperature overnight. All of the volatiles were removed under reduced pressure and water (40 mL) was added. The suspension was extracted with EtOAc (2 x 100 mL). The organic phase was washed with brine, dried over Na₂S0₄, filtered and evaporated to give the crude compound SP-0011379-019 as colorless oil (5.0 g of the crude). LC-MS 249 (M+Na)⁺, purity 96% (UV 214 nm).

To a solution of compound SP-0011379-019 (5.0 g, 20 mmol) in DCM (150 mL) were added PCC (8.64 g, 40.0 mmol) and silica gel (5.0 g) at room temperature. The mixture was stirred at room temperature overnight. The mixture was filtered off and the filtrate was evaporated to give a residue. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 2:1) to give the desired compound SP-0011379-022 as a white solid (4.4 g, yield: 89% in two steps). LC-MS 248 (M+H)⁺, purity 80% (UV 214 nm).

To a solution of SP-0011379-022 (1.32 g, 5.36 mmol) in MeOH.NH₃ (1M, 3.0 mL) was added l-methyl-3,5-dinitropyridin-2(lH)-one (1.6 g, 8.0 mmol) slowly at room temperature. The mixture was stirred at 90 °C for 0.5 h under microwave irridation. The volatiles were removed and the residue was dissolved in DCM (100 mL). The organic layer was washed with saturated aqueous Na₂C0₃ (20 mL), brine (20 mL) and concentrated under reduced pressure to give a crude product. The crude product was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 3: 1) to give the desired compound SP-0011379-028 A (1.2 g) and SP-0011379-028 B (0.8 g) as a yellow solid (yield: 61%). LC-MS 328 (M+H)⁺, purity 84% (UV 214 nm).

A mixture of SP-0011379-028 A (250 mg, 0.76 mmol) and Pd/C (100 mg, 10%) in MeOH (20 mL) was stirred at room temperature overnight under H₂ atmosphere. The resulting mixture was filtered through celite and the filtrate was evaporated to give the crude product SP-0011379-037 as oil (150 mg of the crude) which was used directly in the next step of reaction without purification. LC-MS 164 (M+H)⁺, purity 92% (UV 214 nm).

To a solution of 4-chloro-6-fluoroquinazoline (279 mg, 1.533 mmol) in isopropanol (10 mL) were SP-0011379-037 (250 mg, 1.533 mmol) and triethylamine (0.8 mL). The mixture was stirred at 70 °C for 8 h. The mixture was cooled down and isopropanol was removed by rotary evaporation to give a residue. The residue was purified by silica gel column chromatography (using EtOH/EtOAc = 0: 100 - 9:91) to give the title compound A32-001 (45 mg, yield: 9.5%). LC-MS 295 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6): δ 8.47 (s, 1 H), 8.07-8.04 (m, 1 H), 7.83-7.77 (m, 2 H), 7.66-7.61 (m, 1 H), 6.94 (d, J = 2.4 Hz, 1 H), 4.73-4.64 (m, 1 H), 3.30-3.24 (m, 1 H), 2.97-2.86 (m, 3 H), 2.31-2.26 (m, 1 H), 1.95-1.85 (m, 1 H). Example 163: Preparation of compound A32-002 & A32-003

SP-0010418-162-1 SP-0010418-162-2 SP-0010418-162-3A

SP-0010418-162-4A SP-0010418-162-4B A32-003 A32-002

To a solution of 2-cyclopenten-l-one (2.46 g, 30.0 mmol), benzyl carbamate (5.436 g, 36.0 mmol), Bi(N0₃)₃.5H₂0 (2.18g, 4.5 mmol) in DCM (6 mL) was stirred at room temperature for 20 h. The mixture was quenched with saturated NaHC03 (50 mL) and extracted DCM (2 x 50 mL).The organic layer was dried over Na₂S0₄, filtered and evaporated to give a residue. The residue was purified by silica gel column chromatography (using petroleum ether/EtOAc = 10:0 - 7:3) to give compound SP-0010418-2 (3.5 g, yield: 51%). LC-MS 234 (M+H)⁺, 95% (UV 214 nm).

A mixture of SP-0010418-2 (1.2 g, 5.128 mmol), l-methyl-3,5-dinitropyridin- 2(lH)-one (1.53 g, 7.692 mmol) and 7.0 M NH₃/MeOH (2.0 mL) in MeOH (8.0 mL) was stirred at 90 °C for 0.5 h under microwave irridation. The residue was purified by silica gel column chromatography (using petroleum ether/EtOAc = 10:0 - 7:3) to give mixture SP- 0010418-162-3A and SP-0010418-162-3B (910 mg, yield: 57%). LC-MS 314 (M+H)⁺, 91% (UV 214 nm).

A mixture of SP-0010418-162-3A and SP-0010418-162-3B (910 mg, 2.907 mmol) and 10% Pd/C (100 mg) in EtOH (10 mL) was stirred under H₂ (1 atm) at room temperature for 18 h. The mixture was filtered through a pad of Celite. The filtrate was evaporated to give a crude mixture SP-0010418-162-4A and SP-0010418-162-4B which was used in next step of reaction without further purification (390 mg, yield: 90%). LC-MS 150 (M+H)⁺, 90% (UV 214 nm).

To a solution of 4-chloro-6-fluoroquinazoline (303 mg, 1.667 mmol) in isopropanol (10 mL) was added the mixture of SP-0010418-162-4A and SP-0010418-162-4B (250 mg, 1.667 mmol) and triethylamine (0.8 mL). The resulting mixture was stirred at 70 °C for 5 h. The mixture was cooled down and isopropanol was removed by rotary evaporation to give a residue. The residue was purified by silica gel column chromatography (using EtOH/EtOAc = 0: 100 - 1 : 10) to give the title compound A32-002 (35 mg, yield: 7.1%). LC-MS 295 (M+H)⁺, purity 97% (UV 214 nm); 1H NMR (400 MHz, CD30D-d4): δ 8.41 (s, 1 H), 7.90- 7.87 (m, 1 H), 7.75-7.67 (m, 2 H), 7.55-7.50 (m, 1 H), 6.7 (d, J =2.0 Hz, 1 H), 5.92 (t, J = 8.0 Hz, 1 H), 2.97-2.90 (m, 1 H), 2.85-2.76 (m, 1 H), 2.64-2.55 (m, 1 H), 2.10-2.01 (m, 1 H). A32-003 (12 mg, yield: 2.4%). LC-MS 295 (M+H)+, purity 94% (UV 214 nm); 1H NMR (400 MHz, DMSO-d6): δ 8.39 (s, 1 H), 7.92-7.89 (m, 1 H), 7.68-7.65 (m, 2 H), 7.53- 7.47 (m, 1 H), 6.93 (d, J = 2.0 Hz, 1 H), 5.06-4.98 (m, 1 H), 3.35-3.23 (m, 2 H), 2.95-2.87 (m, 2 H).

Example 164: Preparation of compound A32-004

A32-002 A32-004

A mixture of 2-hydroxyacetic acid (62 mg, 0.814 mmol), HATU (340 mg, 0.895 mmol), DIE A (0.5 mL) and A32-002 (120 mg, 0.407mmol) in DMF (5 mL) was stirred at 50 °C for 48 h. The mixture was cooled down, quenched with water (10 mL), and extracted with EtOAc (2 x 20 mL). The organic layer was dried over Na₂S0₄, filtered and evaporated to give a residue. The residue was purified by silica gel column chromatography (using MeOH/EtOAc = 0: 100 - 9:91) to give the title compound A32-004 (30 mg, yield: 21%). LC-MS 380 (M+H)⁺, purity 92% (UV 214 nm); 1H NMR (400 MHz, CD30D-d4): δ 8.58- 8.53 (m, 2 H), 8.07-8.01 (m, 2 H), 7.81-7.77 (m, 1 H), 7.66-7.63 (m, 1 H), 5.22 (t, J = 5.6 HZ, 1H), 4.17 (s, 2 H), 3.60-3.50 (m, 2 H), 3.21-3.13 (m, 2 H).

Example 165: Preparation of compound A32-005

A32-002 A32-005

A mixture of 1-hydroxycyclopropanecarboxylic acid (104 mg, 1.017 mmol), HATU

(425 mg, 1.118 mmol), DIEA (0.5 mL) and DMAP (12 mg) in DMF/THF (4 mL / 4 mL) was stirred at room temperature for 0.5 h. To this mixture was added A32-002 (150 mg, 0.508 mmol). The resulting mixture was stirred at 50 °C for 18 h. The mixture was cooled down and quenched with water (10 mL), and extracted with EtOAc (2 x 20 mL). The organic layer was dried over Na₂S0₄, filtered and evaporated to give a residue. The residue was purified by silica gel column chromatography to give the title compound A32-005 (90 mg, yield: 46%). LC-MS 380 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (400 MHz, CD₃OD-d4): δ 8.68-8.60 (m, 2 H), 8.16-8.07 (m, 2 H), 7.85-7.72 (m, 2 H), 5.34-5.31 (m, 1 H), 3.62-3.52 (m, 2 H), 3.24-3.16 (m, 2 H), 1.34-1.31 (m, 2 H), 1.12-1.09 (m, 2 H).

Example 166: Preparation of compound A32-006

A32-001 A32-006

To a solution of compound A32-001 (155 mg, 0.5 mmol) in DCM (10 mL) cooled in an ice-bath were added acetic anhydride (260 mg, 2.5 mmol) and TEA (0.5 mL) slowly. The reaction mixture was stirred at room temperature for 3 h. The resulting mixture was evaporated to give a residue. The residue was dissolved in EtOAc (50 mL), washed with brine (10 mL). The organic phase was dried over anhydrous Na₂S0₄, filtered and evaporated to give a crude product. The crude product was purified by silica gel column chromatography (eluting with PE: EtOAc = 1 :0 - 1 : 1) to give the title compound A32-006 as a white solid (70 mg, yield: 40%). LC-MS 352 (M+H)⁺, purity 96% (UV 214 nm); 1H NMR (DMSO-d6, 400 MHz) δ 10.05 (s, 1 H), 8.48 (s, 1 H), 8.45 (s, 1 H), 8.26-8.23 (dd, Ji = 2.8 Hz, h = 10.4 Hz, 1 H), 8.08 (d, J = 7.2 Hz, 1 H), 7.83-7.66 (m, [1+1+1] H), 4.63-4.60 (m, 1 H), 3.24-3.19 (m, 2 H), 2.96-2.87 (m, [1+2] H), 2.18-2.14 (m, 1 H), 2.06 (s, 3 H), 1.90-1.82 (m, 1 H).

Example 167: Preparation of compound A32-007

A32-007

A mixture of SP-0011379-028-B (400 mg, 1.22 mmol) and Pd/C (100 mg, 10%) in MeOH (20 mL) was stirred at room temperature under H₂ atmosphere overnight. The mixture was filtered through celite and the filtrate was evaporated to give the crude product SP-0011379-001 as pink oil (170 mg of the crude) which was used in the next step of reaction without further purification. LC-MS 164 (M+H)⁺, purity 43% (UV 214 nm).

To a solution of SP-0011379-001 (170 mg, 1.0 mmol) in i-PrOH (10 mL) were added 4-chloro-6-fluoroquinazoline (182 mg, 1.0 mmol) and TEA (0.7 mL). The mixture was stirred at 70 °C for 3.0 h. The resulting mixture was evaporated and the residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 6: 1 - 1 :1) to give the desired compound SP-0011379-005 as a pale yellow solid (110 mg, yield: 35%>). LC-MS 310 (M+H)⁺, purity 62% (UV 214 nm).

To a solution of SP-0011379-005 (110 mg, 0.35 mmol) in DCM (10 mL) was added acetyl chloride (54 mg, 0.70 mmol) slowly. The reaction mixture was cooled down in an ice-bath and TEA (0.2 mL) was added in. The resulting mixture was stirred at room temperature for 3.0 h. The reaction mixture was evaporated to give a rresidue and the residue was diluted with EtOAc (50 mL). The organic phase was washed with brine (10 mL), dried over anhydrous Na₂S0₄, filtered and concentrated to give a crude product. The crude product was purified by Pre-HPLC to provide the title compound compound A32-007 as a white solid (75 mg, yield: 61%). LC-MS 352 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (DMSO-d₆, 400 MHz) δ 9.95 (s, 1 H), 8.69 (s, 1 H), 8.51 (s, 1 H), 8.47 (d, J = 8.4 Hz, 1 H), 8.21-8.17 (dd, Ji = 2.8 Hz, J₂ = 10.0 Hz, 1 H), 8.82-8.78 (dd, Ji = 5.6 Hz, J₂ = 9.2 Hz, 1 H), 7.73-7.68 (m, [1+1] H), 5.76-5.73 (m, 1 H), 2.87-2.84 (m, 2 H), 2.08-2.03 (m, 2 H), 1.96 (s, 3 H), 1.90-1.85 (m, 2 H). Example 168: Preparation of compound A32-008

A32-001 A32-008

To a solution of compound A32-001 (155 mg, 0.5 mmol), 2-hydroxy-2- methylpropanoic acid (118 mg, 1.0 mmol) in dry DMF (4.0 mL) were added HATU (286 mg, 0.75 mmol) and TEA (0.4 mL). The mixture was stirred at room temperature overnight. The resulting mixture was quenched by water (20 mL) and extracted with EtOAc (2 x 50 mL). The organic phase was washed with water (20 mL), brine (20 mL) and dried over anhydrous Na₂S0₄, filtered and concentrated to give a residue. The residue was purified by Pre-HPLC to provide the title compound A32-008 (45 mg, yield: 22%) as a white solid.

LC-MS 410 (M+H)⁺, purity 97% (UV 214 nm) ; 1H NMR (DMSO-d₆, 400 MHz) δ 9.95 (s, 1 H), 8.49 (s, 1 H), 8.46 (s, 1 H), 8.26-8.23 (dd, Ji = 2.8 Hz, J₂ = 10.0 Hz, 1 H), 8.09 (d, J = 7.2 Hz, 1 H), 7.86 (d, J = 2.4 Hz, 1 H), 7.79-7.66 (m, [1+1] H), 4.73 (s, 1 H), 4.64-4.62 (m, 1 H), 3.25-3.19 (m, 1 H), 2.97-2.88 (m, [1+2] H), 2.51-2.50 (m, 2 H), 2.16-2.14 (m, 1 H), 1.90-1.84 (m, 1 H), 1.23 (s, [3+3] H).

Example 169: Preparation of compound A32-009

A32-001 A32-009

To a solution of compound A32-001 (155 mg, 0.5 mmol) in DCM (10 mL) cooled in an ice-bath were added methanesulfonyl chloride (114 mg, 1.0 mmol) and TEA (0.4 mL) slowly. The mixture was stirred at room temperature for 3 h. The resulting mixture was evaporated directly to give residue and the residue was dissolved in EtOAc (50 mL). The organic phase was washed with brine (10 mL), dried over anhydrous Na₂S04, filtered and concentrated to give a crude product. The crude product was purified by silica gel chromatography (PE: EtOAc = 10—1 : 1) to give the compound A32-009 as a white solid (60 mg, yield: 34%). LC-MS 388 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (DMSO-d6, 400 MHz) δ 9.82 (s, 1 H), 8.48 (s, 1 H), 8.45 (s, 1 H), 8.26-8.20 (m, [1+1] H), 8.10 (d, J = 7.2 Hz, 1 H), 7.79-7.66 (m, [1+1] H), 7.36 (s, 1 H), 4.65-4.60 (m, 1 H), 3.27-3.21 (m, 1 H), 3.01 (s, 3 H), 2.98-2.87 (m, [1+2] H), 2.18-2.07 (m, 1 H), 1.90-1.82 (m, 1 H).

Example 170: Preparation of compound A32-010 and A32-011

SP-0011379-037

To a solution of SP-0011379-037 (150 mg, 0.92 mmol) in i-PrOH (10 mL) were added 4,7-dichloroquinazoline (199 mg, 1.0 mmol) and TEA (1.0 mL). The mixture was stirred at 70 °C for 3 h. All of the volatiles were evaporated to give a residue. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 6: 1 - 1 : 1) to give the desired compound A32-010 as a white solid (80 mg, yield: 24%). LC-MS 326 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (DMSO-d6, 400 MHz) δ 8.49 (s, 1 H), 8.41 (d, J = 9.2 Hz, 1 H), 8.28-8.25 (m, 1 H), 7.75-7.72 (m, [1+1] H), 7.59-7.56 (dd, Ji =

2.4 Hz, h = 8.8 Hz, 1 H), 6.68 (d, J = 2.4 Hz, 1 H), 5.05 (s, 2 H), 4.58-4.54 (m, 1 H), 3.10-

3.05 (m, 1 H), 2.86-2.75 (m, [1+2] H), 2.14-2.10 (m, 1 H), 1.80-1.75 (m, 1 H).

To a solution of compound A32-010 (50 mg, 0.15 mmol) in DCM (10 mL) cooled in an ice-bath were added acetic anhydride (76 mg, 0.75 mmol) and TEA (0.3 mL). The mixture was stirred at room temperature for 20 h. The resulting mixture was evaporated directly and the residue was dissolved in EtOAc (50 mL). The organic phase was washed with brine (10 mL) and dried over anhydrous Na₂S0₄, filtered and concentrated to give a crude product. The crude product was purified by silica gel column chromatography (using petroleum ether: EtOAc = 6: 1 - 1 : 1) to give the title compound A32-011 as a white solid (30 mg, yield: 54%). LC-MS 368 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (DMSO- d6, 400 MHz) δ 10.05 (s, 1 H), 8.50 (s, 1 H), 8.45-8.40 (m, [1+1] H), 8.29 (d, J = 7.6 Hz, 1 H), 7.82 (s, 1 H), 7.73 (s, 1 H), 7.59-7.57 (dd, Ji = 2.0 Hz, J₂ = 8.8 Hz, 1 H), 4.64-4.61 (m, 1 H), 3.23-3.17 (m, 1 H), 2.97-2.87 (m, [1+2] H), 2.17-2.15 (m, 1 H), 2.07 (s, 3 H), 1. SOUS (m, 1 H). Example 171: Preparation of compound A32-013

SP-0010529-036 SP-0010529-054 SP-0010529-075

To a suspension of potassium tert-butanolate (5.37 g, 48 mmol) in dry ethyl ether (100 mL) at 0 °C was added 3,4-dihydronaphthalen-l(2H)-one (5.74 g, 40 mmol) dropwise. The mixture was stirred at 0 °C for 0.5 h. Then To this resulting mixture was added t-butyl nitrite (6.18 g, 60 mmol) slowly. The reaction mixture was stirred for additional 1 h. The mixture was poured into cold water (300 mL) and extracted with EtOAc (1000 mL). The organic layer was dried over anhydrous Na₂S04, filtered and concentrated to give a residue. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 3 : 1) to give the desired compound SP-0010529-036 (a mixture of two isomers), (6.0 g, yield: 85%) as a pale green solid. LC-MS 176 (M+H)⁺, purity 97% (UV 214 nm).

A mixture of SP-0010529-036 (4.0 g, 22.8 mmol), Pd/C (400 mg, 10%) and (Boc)₂0 (7.4 g, 34.2 mmol) in MeOH / AcOH (100 mL / 2.0 mL) was stirred at room temperature under H₂ atmosphere overnight. The resulting mixture was filtered off and the filtrate was evaporated to give the crude product which was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10: 1 - 2: 1) to give the desired compound SP-0010529-054 (3.6 g, yield: 60%) as a grey color solid. LC-MS 286 (M+Na)⁺, purity 88% (UV 214 nm).

A mixture of SP-0010529-054 (1.05 g, 4.0 mmol) in 1 , 4-dioxane/HCl (4 M, 20 mL) was stirred at room temperature for 24 h. All of the volatiles were removed under reduced pressure to give the crude product SP-0010529-075 (1.1 g of the crude) as a light red solid which was used in the next of reaction without further purification. LC-MS 164.1 (M+H)⁺, purity 50% (UV 214 nm).

To a solution of compound SP-0010529-075 (580 mg, 2.0 mmol) in i-PrOH (20 mL) were added 4-chloro-6-iodoquinazoline (580 mg, 2.0 mmol) and TEA (1.5 mL). The mixture was stirred at 60 °C for 2 h. The resulting mixture was evaporated to give a residue. The residue was purified by silica gel column chromatography with (using petroleum ether: EtOAc = 5: 1 - 1 : 1) to give the desired compound SP-0010529-079 as light brown solid (400 mg, yield: 48%). LC-MS 418 (M+H)⁺, purity 96% (UV 214 nm). 1H NMR (DMSO- d₆, 400 MHz) δ 8.81 (s, 1 H), 8.48 (s, 1 H), 8.22-8.20 (m, 1 H), 8.04-8.01 (m, 1 H), 7.54- 7.45 (m, 2 H), 7.22-7.10 (m, 3 H), 5.52-5.50 (m, 1 H), 4.85-4.81 (m, 1 H), 4.47-4.45 (m, 1 H), 2.93-2.83 (m, 2 H), 2.16-2.12 (m, 1 H), 1.84-1.79 (m, 1 H).

To a solution of compound SP-0010529-079 (208 mg, 0.5 mmol) in DCM (20 mL) at room temperature was added PCC (540 mg, 2.5 mmol) slowly. The mixture was stirred at room temperature overnight. The mixture was filtered off and the filtrate was evaporated to give a residue. The residue was purified by pre-HPLC to provide the desired compound A30-002 as a brown color solid (230 mg, yield: 90%). LC-MS 416 (M+H)⁺, purity 74% (UV 214 nm).

To a solution of compound A30-002 (124 mg, 0.3 mmol) in dry DMSO (5.0 mL) were added Cul (11 mg, 0.06 mmol) and sodium methanesulfmate (177 mg, 1.5 mmol). The mixture was stirred at 100 °C for 5 h under N₂ atmosphere. The resulting mixture was quenched by water (20 mL), extracted with EtOAc (2 x 50 mL) and washed with brine (20 mL). The organic layer was dried over anhydrous Na₂S0₄, filtered and concentrated to give a residue. The residue was purified by silica gel column chromatography (using petroleum ether : EtOAc = 10:1 - 1 : 1) to give the title compound A32-013 (15 mg, yield: 10%>) as a pale yellow solid. LC-MS 368 (M+H)⁺, purity 96% (UV 214 nm); 1H NMR (DMSO-d₆, 400 MHz) δ 9.12 (d, J = 8.0 Hz, 1 H), 9.04 (s, 1 H), 8.55 (s, 1 H), 8.26-8.23 (dd, Ji = 2.0 Hz, h = 8.8 Hz , 1 H), 7.93-7.90 (m, 2 H), 7.64-7.60 (m, 1 H), 7.44-7.39 (m, 2 H), 7.43- 7.40 (m, 1 H), 3.29 (s, 3 H), 3.12-3.07 (m, 1 H), 2.41-2.37 (m, 2 H).

Example 172: Preparation of compound A33-001

SP-0011273-004 SP-0011273-005 SP-0011273-018

A33-001

A mixture of SP-0011273-004 (2.0 g, 9.4 mmol) in EtOH (10 mL) and hydro- hydrazine (0.95 g, 18.8 mmol) was stirred at room temperature for 16 h , The resulting mixture was evaporated under reduced pressure to give a residue, The residue was purified by silica gel column chromatography (DCM: MeOH = 20: 1) to give the desired product SP- 0011273-005 as a yellow solid (1.0 g, yield: 70%). LC-MS 180 (M+H)⁺, purity 57% (UV 214 nm).

A solution of SP-0011273-005 (1.0 g, 6.7 mmol) in aqueous HC1 (6 N, 20 mL) was stirred at 100 °C for 6 h, cooled down and concentrated to give the desired product SP- 0011273-018 as a brown solid (0.8 g, yield: 69%). LC-MS 138 (M+H)⁺, purity 100% (UV 214 nm).

A mixture of 4-chloro-6-fluoroquinazoline (218.0 mg, 1.2 mmol), SP-0011273-018 (137.0 mg, 1.0 mmol) and triethylamine (0.4 ml) in i-PrOH (4.0 ml) was stirred at 75 °C for 3 h. The resulting mixture was evaporated under reduced pressure to give a residue. The residue was purified by pre-HPLC to give the desired product A33-001 as a white solid (0.10 g, yield: 47%). LC-MS 284 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (DMSO, 400 MHz) δ 12.36 (s, 1 H), 8.25-8.22 (m, 1 H), 8.05-8.04 (m, 1 H), 7.83-7.43 (m, 2 H), 7.42 (s, 0.5 H), 7.23 (s, 0.5 H),4.49 (d, J = 6.4 Hz, 1 H), 2.98-2.97 (m, 1 H), 2.82-2.77 (m, 2 H), 2.73 -2.69 (m, 1 H), 2.16 -2.14 (m, 1 H), 1.89-1.84 (m, 1 H).

Example 173: Preparation of compound A33-002

A mixture of N-(4-oxocyclohexyl) acetamide (6.0 g, 38.5 mmol), DMF-DMA (32.0 g) and TEA (1.0 mL) in toluene (150 mL) was stirred at 110 °C overnight. The resulting mixture was evaporated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (from petroleum ether : EtOAc = 1 : 1 to DCM: MeOH = 10: 1) to give the desired product SP-0011273-004 as a yellow solid (5.0 g, yield: 70%). LC-MS 211(M+H)⁺, purity 70% (UV 214 nm).

To a solution of SP-0011273-004 (2.6 g, 12 mmol) in EtOH (10 mL) were added hydrochloric guanidine (2.3 g, 24 mmol) and EtONa (30 mL). The mixture was stirred at 80 °C for 3 h. The resulting mixture was evaporated under reduced pressure to give the desired product SP-0011273-010 as a yellow solid (3.0 g, yield: 90%). LC-MS 207 (M+H)⁺, purity 80% (UV 214 nm).

A mixture of SP-0011273-010 (2.0 g, 9.6 mmol) and aquesous NaOH (4 N, 20 mL) in MeOH(7.0 mL) was stirred at 100 °C for 16 h. The reaction mixture was cooled down, diluted with water, extracted with DCM (7 x 50 mL). The organic layer was washed with brine (50 mL), dried over Na₂S0₄, concentrated to give the target product SP-0011273-019 as brown oil (586 mg, yield: 39%). LC-MS 165(M+H)⁺, purity 72% (UV 214 nm).

To a solution of 4-chloro-6-fluoroquinazoline (138.0 mg, 1.15 mmol) in i-PrOH (2.0 mL) were added SP-0011273-019 (110.0 mg, 0.63 mmol) and TEA (0.5 mL). The mixture was stirred at 75 °C for 3 h. The resulting mixture was evaporated under reduced pressure to give a residue. The residue was purified by pre-HPLC to give the desired product A33- 002 as a white solid (80.0 mg, yield: 48%). LC-MS 311(M+H)⁺, purity 100% (UV 214 nm); 1H NMR (CDC1₃, 400 MHz) δ 8.59 (s, 1 H), 8.01 (s, 1 H), 7.83-7.79 (m, 1 H), 7.47- 7.42 (m, 1 H), 7.23-7.19 (m, 1 H), 5.37-5.35 (m, 1 H), 4.82 (s, 1 H), 4.66 (d, J = 6.4 Hz, 1 H), 3.18-3.13 (m, 1 H), 2.88-2.88 (m, 2 H), 2.62 -2.56 (m, 1 H), 2.25 -2.23 (m, 1 H), 1.99 - 1.96 (m, 1 H), 1.95-1.94 (m, 1 H).

Example 174: Preparation of compound A33-003

SP-0011273-038 A33-003

To a solution of SP-0011273-004 (1.5 g, 7.2 mmol) in EtOH (7 mL) were added acetimidamide (1.4 g, 14.4 mmol) and EtONa (20 mL). The mixture was stirred at 80 °C for 3 h. The resulting mixture was evaporated under reduced pressure to give the product SP- 0011273-026 as a yellow solid (1.7 g, yield: 90%). LC-MS 206 (M+H)⁺, purity 76% (UV 214 nm).

A mixture of SP-0011273-026 (1.7 g, 9.6 mmol) and aqueous NaOH (4 M, 15 mL) in MeOH (5 mL) was stirred at 100 °C for 16 h. The reaction mixture was cooled down, diluted with water and extracted with DCM (7 x 50 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated under reduced pressure to give the product SP-0011273- 038 as brown oil (0.6 g, yield: 45%). LC-MS 164 (M+H)⁺, purity 90% (UV 214 nm).

To a solution of 4-chloro-6-fluoroquinazoline (0.11 g, 0.6 mmol) in i-PrOH (2.0 ml) were added SP-0011273-038 (81 mg, 0.5 mmol) and TEA (0.25 mL). The mixture was stirred at 75 °C for 3 h. The resulting mixture was evaporated under reduced pressure to give a residue. The residue was purified by pre-HPLC to give the title compound A33-003 as a white solid (61.0 mg, yield: 45%). LC-MS 310 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (CDCls, 400 MHz) δ 8.59 (s, 1 H), 8.30 (s, 1 H), 7.84-7.80 (m, 1 H), 7.48-7.43 (m, 1 H), 7.25-7.19 (m, 1 H), 5.39-5.38 (m, 1 H), 4.68 (m, 1 H), 3.32-3.27 (m, 1 H), 3.04-2.99 (m, 2 H), 2.74 -2.67 (m, 1 H), 2.63 (s, 3 H), 2.31 -2.30 (m, 1 H), 2.03 -1.99 (m, 1 H).

Example 175: Preparation of compound A33-004

SP-0011273-039 A33-004

To a solution of SP-0011273-004 (0.5 g, 7.2 mmol) in EtOH (4 mL) were added acetimidamide (0.4 g,4.8 mmol) and EtONa (15 mL). The mixture was stirred at 80 °C for 3 h. The resulting mixture was evaporated under reduced pressure to give the product SP- 0011273-027 as a yellow solid (0.6 g, yield 90%). LC-MS 192 (M+H)⁺, purity 55% (UV 214 nm).

To a solution of SP-0011273-027 (0.6 g, 3.2 mmol) in MeOH (5 mL) was added aqueous NaOH (4 N, 15 mL). The mixture was stirred at 100 °C for 16 h. The reaction mixture was cooled down, diluted with water (5 mL) and extracted with DCM (4 x 50 mL). The organic phase was dried over Na₂S0₄, filtered and concentrated to give the product SP- 0011273-039 as brown oil (0.13 g, yield: 35%). LC-MS 150 (M+H)⁺, purity 91% (UV 214 nm).

To a solution of 4-chloro-6-fluoroquinazoline (95.0 mg, 0.5 mmol) in i-PrOH (2.0 mL) were added SP-0011273-039 (65.0 mg, 0.42 mmol) and TEA (0.25 mL). The mixture was stirred at 75 °C for 3 h. The resulting mixture was evaporated under reduced pressure to give a residue. The residue was purified by pre-HPLC to give the product A33-004 as a white solid (50.0 mg, yield: 46%). LC-MS 296 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (CDCI3, 400 MHz) δ 8.95 (s, 1 H), 8.60 (s, 1 H), 8.40 (s, 1 H), 7.84-7.81 (m, 1 H), 7.48-7.43 (m, 1 H), 7.26-7.19 (m, 1 H), 5.42-5.40 (m, 1 H), 4.74-4.71 (m, 1 H), 3.39-3.34 (m, 1 H), 3.10-2.06 (m, 2 H), 2.79-2.72 (m, 1 H), 2.10 (m, 1 H), 2.02 (m, 1 H).

Exampl 176: Preparation of compound A33-005

SP-0011273-018 A33-005

To a solution of 4,6-dichloroquinazoline (140 mg, 0.7 mmol) in i-PrOH (3.0 mL) were added 4,5,6,7-tetrahydro-2H-indazol-5-amine (80 mg, 0.575 mmol) and TEA (0.3mL). The mixture was stirred at 75 °C for 3 h. The resulting mixture was evaporated under reduced pressure to give a residue. The residue was purified by pre-HPLC to give the product A33-005 as a white solid (90 mg, yield: 54%). LC-MS 301 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (CDC1₃, 400 MHz) δ 8.59 (s, 1 H), 7.73-7.71 (m, 1 H), 7.61-7.59 (m, 1 H), 7.54-7.53 (m, 1 H), 7.34 (s, 1 H), 5.54-5.52 (m, 1 H), 4.74-4.70 (m, 1 H), 3.12- 3.07 (m, 1 H), 2.85-2.79 (m, 2 H), 2.62-2.56 (m, 1 H), 2.170-2.06 (m, 1 H).

Example 177: Preparation of compound A33-006

To a solution of 4,6-dichloroquinazoline (146.0 mg, 0.732 mmol) in i-PrOH (4.0 mL) were added SP-0011273-019 (O.lg, 0.61 mmol) and TEA (0.3ml). The mixture was stirred at 75 °C for 3 h. The resulting mixture was evaporated under reduced pressure to give a residue. The residue was purified by pre-HPLC to give the product A33-006 as a brown solid (0.10 mg, yield: 48%). LC-MS 328 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR (DMSO, 400 MHz) δ 8.53-8.50 (m, 2 H), 8.22-8.20 (m, 1 H), 7.99 (s, 1 H), 7.80-7.78 (m, 2 H), 7.72-7.69 (m, 1 H), 6.32 (s, 1 H), 4.58-4.51 (m, 1 H), 3.00-2.95 (m, 1 H), 2.77- 2.73 (m, 2 H), 2.67 -2.60 (m, 1 H), 2.12-2.11 (m, 1 H), 1.95 -1.89 (m, 1 H). Example 178: Preparation of compound A33-007

SP-0011273-066 A33-007

A mixture of SP-0011273-010 (3.0 g,14.96 mmol) in aqueous HCI (6 mL) at -15 °C was added an ice-cooled solution of NaN0₂ (2.1 g, 29.9 mmol) in water (5 mL) dropwise. The mixture was stirred at -15 °C for 1 h. The reaction mixture was neutralized to pH = 7 with aqueous NaOH (30%) and extracted with EtOAc (4 x 50 mL). The organic layer was washed with brine (100 mL), dried over Na₂S0₄, filtered and concentrated to give the product SP-0011273-061 as brown oil (0.85g, yield: 31%). LC-MS 226(M+H)⁺, purity 100% (UV 214 nm).

A mixture of SP-0011273-061 (0.85 g, 3.6 mmol) and 2-(ethylamino)ethanol (3 mL, 33 mmol) in DMF (3 mL) was stirred at 80 °C for 4 h. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (4 x 50 mL). The organic layer was washed with brine (150 mL), dried over Na₂S0₄, filtered and concentrated to give the product SP- 0011273-062 as a brown oil (1.0 g, yield: 80%). LC-MS 279(M+H)⁺, purity 82% (UV 214 nm).

A mixture of SP-0011273-062 (1.0 g, 35.1mmol) and aqueous NaOH (4 N, 20 mL) in MeOH (5 mL) was stirred at 100 °C for 16 h. The reaction mixture was cooled down, diluted with water and extracted with DCM (4 x 50 mL). The organic layer was washed with brine (150 mL), dried over Na₂S0₄, filtered and concentrated to provide the product SP-0011273-066 as brown oil (0.20 g, yield: 30%). LC-MS 237(M+H)⁺, purity 65% (UV 214 nm).

To a solution of 4-chloro-6-fluoroquinazoline (0.11 mg, 0.58 mmol) in i-PrOH (3.0 mL) were added SP-0011273-066 (90 mg, 0.383 mmol) and TEA (0.3 mL). The mixture was stirred at 75 °C for 3 h. The resulting mixture was evaporated under reduced pressure to give a residue. The residue was purified by pre-HPLC to give the target product A33-007 as a white solid (0.10 g, yield: 54%). LC-MS 383 (M+H)⁺, purity 95% (UV 214 nm); 1H NMR (CDCls, 400 MHz) δ 8.65 (s, 1 H), 8.04 (s, 1 H), 7.91-7.88 (m, 1 H), 7.54-7.49 (m, 1 H), 7.38-7.35 (m, 1 H), 5.70-5.68 (m, 1 H), 4.73-4.68 (m, 1 H), 3.90-3.87 (m, 1 H), 3.73- 3.65 (m, 4 H), 3.20-3.14 (m, 1 H), 2.92-2.82 (m, 2 H), 2.66-2.60 (m, 1 H), 2.30-2.26 (m, 1 H), 2.06-1.99 (m, 1 H).

eparation of compound A33-008 4M NaOH, MeOH reflux, 16 h

SP-0011273-009 A33-008

To a solution of SP-0011273-004 (2.5 g, 11.7 mmol) in EtOH (10 mL) were added glycine (0.95 g, 18.8 mmol) and KOH (0.65 g, 11.7 mmol). The mixture was stirred at 80 °C for 2 h , The resulting mixture was evaporated under reduced pressure to give a residue, The residue was dissolved in Ac₂0 (15 mL), and stirred at 100 °C for additional 1 h. The reaction mixture was cooled down, diluted with aquesous NaHC0₃ (150 mL) and extracted with EtOAc (3 x 50 mL). The combined organic phase was washed with brine (100 mL), dried over Na₂S0₄, filtered and concentrated to give a residue. The residue was purified by silica gel column chromatography with (DCM : MeOH = 30: 1) to give the target product SP-0011273-006 as yellow oil (2.0 g, yield: 70%). LC-MS 221 (M+H)⁺, purity 62% (UV 214 nm).

A mixture of SP-0011273-006 (2 g, 9.6 mmol) and aqueous NaOH (4 N, 20 mL) in MeOH (7 mL) was stirred at 100 °C for 16 h. The reaction mixture was cooled down, diluted with water and extracted with DCM (7 x 50 mL). The organic layer was washed with brine (20 mL),dried over Na₂S0₄, filtered and concentrated to give the target product SP-0011273-009 as brown oil (386 mg, yield: 35%). LC-MS 137(M+H)⁺, purity 51% (UV 214 nm).

A mixture of 4-chloro-6-fluoroquinazoline (0.55 g, 3.27 mmol), SP-0011273-009 (0.3 mg, 2.2 mmol) and Et₃N (0.9 mL) in i-PrOH (7.0 mL) was stirred at 75 °C for 3 h. The resulting mixture was evaporated under reduced pressure to give a residue. The residue was purified by pre-HPLC to give the target product A33-008 as a brown solid (35.0 mg, yield: 12%). LC-MS 283 (M+H)⁺, purity 100% (UV 214 nm); 1H NMR(DMSO, 400 MHz) δ 10.36 (s, 1 H), 8.49-8.45 (m, 1 H), 8.26-8.23 (m, 1 H), 8.01-7.99 (m, 1 H), 7.77-7.64 (m, 2 H), 6.447-6.441 (m, 1 H), 4.46 (d, J = 6.4 Hz, 1 H), 3.01-2.98 (m, 1 H), 2.74-2.61 (m, 2 H), 2.09 -2.06 (m, 1 H), 1.75-1.68 (m, 1 H).

Example 180: Combination Chemotherapy

Combination studies of A31-001, A31-001 A and A31-00 IB with AC220 or lapatinib on ES-2 or SUM159 cell lines were conducted. In brief, AC220 showed enhanced cytotoxicity towards ES-2 cells in the presence of A31-001, A31- 001A or A31-001B (see Figure 73 for combination of A31-001 A with AC220, and Figure 74 for combination of A31-001B with AC220). This means that we observed a synergistic effect or combination effect between AC220 and one of these three autophagy inhibitors on ES-2 cancer cell line.

INCORPORATION BY REFERENCE

All publications including all U.S. patents and U.S. published patent applications cited herein are hereby incorporated by reference. In addition, the entire contents of U.S. Patent Application Publication Nos. 2010/0267704, 2012/0258975, 2012/0315244, and 2012/0301463, and PCT International Patent Application Publication No. WOl 1/143444 are hereby incorporated by reference.

EQUIVALENTS

While several embodiments of the present invention have been described and illustrated herein, those of ordinary skill in the art will readily envision a variety of other means and/or structures for performing the functions and/or obtaining the results and/or one or more of the advantages described herein, and each of such variations and/or modifications is deemed to be within the scope of the present invention. More generally, those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary and that the actual parameters, dimensions, materials, and/or configurations will depend upon the specific application or applications for which the teachings of the present invention is/are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. It is, therefore, to be understood that the foregoing embodiments are presented by way of example only and that, within the scope of the appended claims and equivalents thereto, the invention may be practiced otherwise than as specifically described and claimed. The present invention is directed to each individual feature, system, article, material, kit, and/or method described herein. In addition, any combination of two or more such features, systems, articles, materials, kits, and/or methods, if such features, systems, articles, materials, kits, and/or methods are not mutually inconsistent, is included within the scope of the present invention.

Claims

A compound represented by Formula IV, Formula V, or Formula VI

Formula VI

wherein, independently for each occurrence,

Y is F, CN, NO2, S0₂R, SO2NR2, NRSO2R, I, CH₃, CI, CF₃, or CONR₂;

-C(OH)(CF₃)₂, NRC(0)NR₂, -OCH₂CH₂-(N-morpholinyl),

six-membered amide ring;

R² may be present on the saturated ring or the unsaturated ring;

m is 0, 1, or 2; and

2. A compound represented by Formula VII

Formula VII

wherein, independently for each occurrence,

Y is F, CN, NO2, S0₂R, SO2NR2, NRSO2R, I, CH₃, CI, CF₃, or CONR₂;

q is 1 or 2;

n is 0, 1, or 2;

R¹ is NRCOR, C0₂R, NRSO2R, SO2 -C(OH)(CF₃)₂,

NRC(0)NR₂, -OCH₂CH₂-(N-morpholinyl),

3. A compound represented by Formula VIII

Formula VIII

wherein, independently for each occurrence,

Y is F, CN, NO2, S0₂R, SO2NR2, NRSO2R, I, CH₃, CI, CF₃, or CONR₂;

n is 0, 1, or 2; and

4. A compound represented by Formula IX

Formula IX

wherein, independently for each occurrence,

taken together, form a five- or six-membered amide ring;

m is 0, 1, or 2;

n is 0, 1, or 2; and

5. A compound of Formula I, Formula II, or Formula III, as depicted in Figure 2, wherein Ri is Y as defined above; R₂-R₇ is H or R² as defined above; n is 0, 1, or 2; and the 2-substituent in Figure 2 is H, halo, CN, N0₂, fluoroalkyl, or alkyl.

6. A compound selected from the group consisting of the compounds listed in Figure

1.

7. A method for inhibiting autophagy in a subject for whom inhibition of autophagy is beneficial, comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-6, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby inhibiting autophagy activity in the subject.

8. A method of treating or preventing cancer, comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1-6, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby treating or preventing cancer.

9. A method of treating or preventing cancer, comprising the step of co-administering to a subject in need thereof (i) a therapeutically effective amount of a compound of any one of claims 1-6, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, and (ii) a therapeutically effective amount of a second active agent, thereby treating or preventing cancer.

10. The method of claim 9, wherein the second active agent is an anti-cancer agent.

11. The method of claim 9, wherein the second active agent is a tyrosine kinase inhibitor or a Raf kinase inhibitor.

12. The method of claim 9, wherein the second active agent is imatinib, gefitinib, erlotinib, sunitinib, sorafenib, or quizartinib (AC220).

13. The method of any one of claims 9-12, wherein the cancer is breast cancer or ovarian cancer.

14. A method of treating or preventing acute pancreatitis, comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1-6, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby treating or preventing pancreatitis.

15. A method of treating or preventing a disease caused by an intracellular pathogen, comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1-6, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby treating or preventing the disease caused by the intracellular pathogen.

16. A method of treating or preventing a lysosomal storage disorder, comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1-6, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby treating or preventing the lysosomal storage disorder.

17. A method of inhibiting autophagy in a cell in need thereof, comprising the step of contacting the cell with a therapeutically effective amount of a compound of any one of claims 1-6, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, thereby inhibiting autophagy in the cell.

18. A method of inactivating a deubiquitinating protease complex, comprising the step of contacting the deubiquitinating protease complex with a compound of any one of claims 1-6, or a pharmaceutically acceptable salt, biologically active metabolite, solvate, hydrate, prodrug, enantiomer or stereoisomer thereof, wherein the deubiquitinating protease complex comprises USP3 and USP10.