RU2805511C2 - Proteolysis modulators and related uses - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to a compound having the chemical structure of ULM-L-PTM, or a pharmaceutically acceptable salt thereof, which can be used to degrade rat Kirsten sarcoma (KRas) protein. In this structure, (a) L is represented by the formula -(A^L)_q-, where q is an integer from 1 to 50; each A^L is independently selected from the group consisting of CR^L1R^L2, O, NR^L3, CO, C≡ C, C_3–11 heterocycloalkyl containing 1 or 2 N atoms, optionally substituted with 1-3 R^L1 and/or R^L2 groups , and phenyl optionally substituted with 1–3 R^L1 and/or R^L2 groups; and R^L1, R^L2 and R^L3 are each independently H or C_1–8 alkyl; (b) ULM is the following fragment; where W³ is selected from , 5–10-membered heteroaryl or phenyl, wherein the heteroaryl contains 1 or 2 atoms independently selected from N, O or S, and wherein the heteroaryl and phenyl are optionally substituted with 1 or 2 C_1-6 alkyls; R₉ and R₁₀ are independently hydrogen or C_1-6 alkyl; R₁₁ is or 5–10 membered heteroaryl optionally substituted with 1 or 2 C_1-6 alkyls, wherein the heteroaryl contains 1 or 2 atoms selected from N, O or S; R₁₂ is either H or C_1-6 alkyl; R₁₃ is either H, C_1-6 alkyl or C_1-6 alkylcarbonyl; each of R_14a, R_14b independently represents either H or C_1-6 alkyl; W⁵ is a 5–10 membered heteroaryl or phenyl, wherein the heteroaryl contains 1 or 2 atoms independently selected from N, O or S, and wherein the heteroaryl and phenyl are optionally substituted with 1 or 2 C_1-6 alkyls; R₁₅ is either H or 5–10 membered heteroaryl optionally substituted with 1 or 2 C_1-6 alkyls, wherein the heteroaryl contains 1 or 2 atoms selected from N, O or S; each R₁₆ is independently either C_1-6 alkyl or C_1-6 alkoxy; o is either 0, 1, 2, 3 or 4; (c) PTM is selected from PTM-III and PTM-IV; where is either phenyl or heterocycloalkyl, wherein the heterocycloalkyl contains 1 N atom; X _PTM is N; R_PTM1A is NR _PTM9 R_PTM10, O-(C_3–6 heterocycloalkyl), wherein the heterocycloalkyl contains 1 N atom, -OC_1–4 alkyl-C_3–6 heterocycloalkyl optionally substituted with 1 or 2 substituents independently selected from C_1–6 alkyl or O-C_1–6 alkyl, wherein the heterocycloalkyl contains either 1 or 2 N atoms, etc.; R_PTM1B is H or; R_PTM9 and R_PTM10 are each independently H, phenyl, or C_1–6 alkyl optionally substituted with either 1 or 2 substituents independently selected from C(O)NH₂, C(O)N(C_1–6 alkyl) ₂ and C(O)N(H)(C_1–6 alkyl); R_PTM2 is H or (C=O)R _PTM2'; R_PTM2' is C_1–6 alkyl, C_2–6 alkene, -N(R_PTM8)₂ or -C(OH)₂; R_PTM3 is either phenyl or naphthalene, each independently substituted with either 1 or 2 substituents independently selected from OH, H and halo; R_PTM4A represents 1 or 2 substituents independently selected from OH, H and halo; R_PTM4B represents either Н or -CH₂CN; R_PTM8 represents H or С_1–6 alkyl; t represents 0, 1, 2, 3, 4, 5, 6; and represents the place of connection of L.

EFFECT: invention also relates to certain compounds, a pharmaceutical composition containing the proposed compounds and their use for treatment of the disease or disorder related to accumulation, aggregation and/or over-activation of KRas.

11 cl, 2 dwg, 14 tbl, 573 ex

Description

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] The present invention claims priority to US Provisional Application No. 62/652676, filed April 4, 2018, entitled "MODULATORS OF PROTEOLYSIS AND ASSOCIATED METHODS OF USE", the contents of which are incorporated herein by reference in their entirety for all purposes.

INCLUSION BY REFERENCE

[0002] US Patent Application Serial No. 15/230,354, filed August 5, 2016, published as Application Publication No. 2017/0065719, and US Patent Application No. 15/206497, filed July 11, 2016, published as publication of US patent application No. 2017/0008904; and US Patent Application 15/209,648, filed July 13, 2016, published as US Patent Application Publication No. 2017/0037004; and US Patent Application No. 15/730728, filed October 11, 2017, published as US Patent Application Publication No. 2018/0099940; and US Patent Application No. 14/686640, filed April 14, 2015, published as US Patent Application Publication No. 2015/0291562; and US Patent Application No. 14/792414, filed July 6, 2015, published as US Patent Application Publication No. 2016/0058872; and US Patent Application No. 14/371956, filed July 11, 2014, published as US Patent Application Publication No. 2014/0356322; and US Patent Application No. 15/074820, filed March 18, 2016, published as US Patent Application Publication No. 2016/0272639; and US Provisional Patent Application No. 15/885671, filed January 31, 2018, published as US Patent Application Publication No. 2018/0215731 A1; and International Patent Application No. PCT/US 2016/023258, filed March 18, 2016, published as International Patent Application Publication No. WO 2016/149668, are incorporated herein by reference in their entirety. In addition, all references cited herein are incorporated herein by reference in their entirety.

TECHNICAL FIELD

[0003] The disclosure provides bifunctional compounds containing a target protein binding moiety and an E3 ubiquitin ligase binding moiety and corresponding methods of use. Bifunctional compounds are useful as targeted ubiquitination modulators, particularly for Kirsten sarcoma ras protein (KRas or KRAS), such as mutant KRas or gain-of-function KRas, which are degraded and/or otherwise inhibited by the bifunctional compounds of the invention.

BACKGROUND OF THE ART

[0004] Most small molecule drugs bind enzymes or receptors in narrow and well-defined pockets. On the other hand, protein-protein interactions are notoriously difficult to target using small molecules due to their large contact surfaces and the shallow grooves or planar surfaces involved. E3 ubiquitin ligases (of which hundreds are known in humans) confer substrate specificity for ubiquitination and are therefore more attractive therapeutic targets than conventional proteasome inhibitors due to their specificity for certain protein substrates. Designing E3 ligase ligands has proven challenging, in part because they must disrupt protein-protein interactions. However, recent developments have provided specific ligands that bind to these ligases. For example, since the discovery of nutlins, the first small molecule E3 ligase inhibitors have been reported; Additional compounds targeting E3 ligases have been reported, but this area remains underdeveloped. For example, since the discovery of nutlins, the first small molecule inhibitors of the murine double minute 2 homolog 2 (MDM2) E3 ligase, additional compounds have been reported that target MDM2 (i.e., human double minute 2 or HDM2) E3 ligases (J. Di, et. al. Current Cancer Drug Targets (2011), 11(8), 987-994).

[0005] One E3 ligase with exciting therapeutic potential is von Hippel-Lindau (VHL) tumor suppressor, the substrate recognition subunit of the E3 VCB ligase complex, which also consists of elongins B and C, Cul2 and Rbx1. The primary substrate of VHL is hypoxia-inducible factor-lα (HIF-lα), a transcription factor that activates genes such as the proangiogenic growth factor VEGF and the red blood cell-inducing cytokine erythropoietin in response to low oxygen levels. The first small molecule von Hippel Lindau ligands (VHL) to the substrate recognition subunit of E3 ligase were generated, and crystal structures were obtained confirming that the compound mimics the binding mode of the transcription factor HIF-1α, a major substrate of VHL.

[0006] Cereblon is a protein that is encoded by the CRBN gene in humans. CRBN orthologs are highly conserved from plants to humans, highlighting its physiological significance. Cereblon forms an E3 ubiquitin ligase complex with damaged DNA binding protein 1 (DDB1), cullin-4A (CUL4A), and regulator of cullin 1 (ROC1). This complex ubiquitinates a number of other proteins. Through a mechanism that is not fully understood, ubquitination of target proteins by cereblons leads to increased levels of fibroblast growth factor 8 (FGF8) and fibroblast growth factor 10 (FGF10). FGF8, in turn, regulates a number of developmental processes, such as the formation of limbs and otic vesicles. Ultimately, this ubiquitin ligase complex is important for limb outgrowth in embryos. In the absence of cereblon, DDB1 forms a complex with DDB2, which functions as a damaged DNA binding protein.

[0007] Inhibitors of apoptosis proteins (IAPs) are a family of proteins involved in the suppression of apoptosis, that is, cell death. The human IAP family contains 8 members, and many other organisms contain IAP homologs. IAPs contain an E3 ligase-specific domain and baculoviral IAP repeat (BIR) domains that recognize substrates and promote their ubiquitination. IAPs promote ubiquitination and can directly bind and inhibit caspases. Caspases are proteases (eg, caspase-3, caspase-7 and caspase-9) that mediate apoptosis. Thus, by binding caspases, IAPs inhibit cell death. However, pro-apoptotic stimuli can lead to the release of the mitochondrial proteins DIABLO (also known as second mitochondrial activator of caspases or SMAC) and HTRA2 (also known as Omi). Binding of DIABLO and HTRA2 appears to block IAP activity.

[0008] SMAC interacts with virtually all known IAPs, including XIAP, c-IAP1, C-IAP2, NIL-IAP, Brace and survivin. The first four amino acids (AVPI) of mature SMAC bind to the IAP moiety, which is thought to be important for blocking the antiapoptotic effects of IAP.

[0009] Bifunctional compounds, such as those described in US Patent Application Publications 2015-0291562 and 2014-0356322 (incorporated herein by reference), function to recruit endogenous proteins to E3 ubiquiin ligase for degradation. In particular, the publications describe bifunctional chimeric compounds or proteolysis-targeted chimeric compounds (PROTACs) that find use as modulators of targeted ubiquitination of a variety of polypeptides and other proteins, which are then degraded and/or otherwise inhibited by the bifunctional compounds.

[0010] The Kirsten rat sarcoma gene (KRAS) is an oncogene encoding KRas, which is a small GTPase signal transduction protein. Ras proteins bind to the plasma membrane and act as switches in transmitting extracellular signals to intracellular responses, thereby regulating, for example, cell division. Numerous activating or gain-of-function mutations are known in the KRas gene, and in fact, KRas is the most commonly mutated gene in cancer. KRas gain-of-function mutations are found in approximately 30% of all human cancers, including, for example, pancreatic cancer (>80%), colon cancer (approximately 40-50%), lung cancer (approximately 30-50%), non-small cell lung cancer, biliary tract malignancies, endometrial cancer, cervical cancer, bladder cancer, liver cancer, myeloid leukemia and breast cancer. These activating mutations impair the ability of KRas to switch between active and inactive states. Key roles for mutant KRas have been established in the initiation, maintenance, progression and metastasis of various cancers, and mutations are often correlated with poor prognosis and increased resistance to chemotherapy and biological therapies, including, for example, epidermal growth factor receptor-targeted therapies. However, despite its key role and high prevalence rates in cancer, the lack of effective treatments that target this oncogene has led to it being considered “untreatable.”

[0011] Thus, there is a continuing need in the art for effective methods of treating a disease associated with overexpression, aggregation, and/or overactivation of KRas (e.g., aggregation of active KRas), such as a gain-of-function mutant of KRas (i.e., KRas , having a gain of function mutation). However, nonspecific effects and the inability to target and modulate mutant KRas remain obstacles to the development of effective treatments. Thus, small molecule therapeutics that target KRas and that enhance or enhance the substrate specificity of VHL, cereblon, MDM2, and IAP would be very useful.

DISCLOSURE OF INVENTION

[0012] The present invention provides bifunctional compounds that function to recruit endogenous proteins to E3 ubiquitin ligase for degradation, and methods of using them. In particular, the present invention provides bifunctional chimeric compounds or proteolysis-targeted chimeric compounds (PROTACs) that find use as modulators of targeted ubiquitination of a variety of polypeptides and other proteins, which are then degraded and/or otherwise inhibited by the bifunctional compounds as described herein . The advantage of the compounds proposed herein is that a wide range of pharmacological activities is possible, compatible with the degradation/inhibition of target polypeptides from virtually any class or family of proteins. Also provided herein are methods of using an effective amount of the compounds as described herein to treat or alleviate a disease condition such as cancer, such as pancreatic cancer, colon cancer, colorectal cancer, lung cancer, non-small cell lung cancer, malignancy. biliary tract, endometrial cancer, cervical cancer, bladder cancer, liver cancer, myeloid leukemia and breast cancer.

[0013] Thus, in one aspect, the invention provides bifunctional compounds or PROTAC compounds that contain an E3 ubiquitin ligase binding moiety (i.e., an E3 ubiquitin ligase ligand or "ULM" group) and a moiety that binds a target protein (i.e. e. a ligand targeting a protein/polypeptide or "PTM" group such that the target protein/polypeptide (e.g. Kirsten rat sarcoma protein (KRas or KRAS) and/or mutant KRas such as KRas ^G12C ) is placed adjacent to the ubiquitin ligase, in order to influence the degradation (and inhibition) of this protein. In a preferred embodiment, the ULM (ubiquitinating ligase modulator) may be a von Hippel-Lindau (VHL) biquitin ligase E3 ligase (VLM) binding fragment, or a cereblon E3 ubiquitin ligase (CLM) binding fragment, or a murine double homologue E3 ubiquitin ligase binding fragment. minutes 2 (MDM2) (MLM), or the E3 ubiquitin ligase IAP binding fragment (i.e., “ILM”). For example, the structure of a bifunctional compound can be depicted as:

[0014] The respective positions of PTM and ULM fragments (eg, VLM, CLM, MLM or ILM), as well as their numbers, as illustrated herein, are offered as examples only and are in no way intended to limit the connections. As will be appreciated by one skilled in the art, the bifunctional compounds described herein can be synthesized in such a way that the number and position of the corresponding functional moieties can be varied as desired.

[0015] In some embodiments, the bifunctional compound further comprises a chemical linker (“L”). In this example, the structure of the bifunctional compound can be depicted as:

wherein PTM is a protein/polypeptide-targeting moiety, L is a linker, e.g., a bond or chemical group linking PTM to ULM, and ULM is an E3 IAP ubiquitin ligase (ILM)-binding moiety or ubiquitin ligase-binding moiety E3 von Hippel-Lindau (VHL) (VLM), or E3 cereblon ubiquitin ligase (CLM) binding fragment, or E3 ubiquitin ligase murine double minute 2 (MDM2) homologue (MDM2) binding fragment (MLM).

[0016] For example, the structure of a bifunctional compound can be depicted as:

where: PTM is a fragment targeting a protein/polypeptide; "L" represents a linker (eg, a bond or chemical linker group) linking PTM and at least one of VLM, CLM, MLM, ILM, or a combination thereof; VLM is a von Hippel-Lindau E3 ubiquitin ligase binding fragment that binds to the E3 ligase VHL; CLM is a cereblon E3 ubiquitin ligase binding fragment that binds to cereblon; MLM is an E3 ubiquitin ligase MDM2 binding fragment that binds MDM2; a ILM is an IAP binding fragment that binds to an IAP.

[0017] In certain preferred embodiments, the ILM is a tetrapeptide fragment of AVPI. Thus, in certain additional embodiments, the ILM of the bifunctional compound comprises the amino acids alanine (A), valine (V), proline (P), and isoleucine (I) or non-natural mimetics thereof, respectively. In further embodiments, the amino acids of the AVPI tetrapeptide moiety are linked to each other by amide bonds (i.e., -C(O)NH- or -NHC(O)-).

[0018] In certain embodiments, compounds as described herein contain multiple independently selected ULMs, multiple PTMs, multiple chemical linkers, or a combination thereof.

[0019] In certain embodiments, the ILM contains chemical moieties such as those described herein.

[0020] In additional embodiments, the VLM may be hydroxyproline or a derivative thereof. Additionally, other proposed VLMs are included in US Patent Application Publication No. 2014/03022523, which, as discussed above, is incorporated herein in its entirety.

[0021] In one embodiment, the CLM contains a chemical group derived from an imide, thioimide, amide, or thioamide. In a specific embodiment, the chemical group is a phthalimido group or an analogue or derivative thereof. In a certain embodiment, the CLM is thalidomide, lenalidomide, pomalidomide, analogs thereof, isosteres thereof, or derivatives thereof. Other proposed CLMs are described in US Patent Application Publication No. 2015/0291562, which is incorporated herein in its entirety.

[0022] In certain embodiments, the MLM may be nutlin or a derivative thereof. In addition, other purported MLMs are included in US Patent Application Publication 15/206497, filed July 11, 2016, published as US Patent Application Publication No. 2017/0008904, which, as discussed above, is incorporated herein in its entirety. In certain additional embodiments, the MLM of the bifunctional compound includes chemical moieties such as substituted imidazolines, substituted spiroindolinones, substituted pyrrolidines, substituted piperidinones, substituted morpholinones, substituted pyrrolopyrimidines, substituted imidazolopyridines, substituted thiazoloimidazolinones, substituted pyrrolopyrrolidinones, and substituted isoquinolinones.

[0023] In additional embodiments, the MLM contains the above core structures with adjacent bis-aryl substitutions positioned as cis or trans configurations.

[0024] In certain embodiments, “L” represents a link. In further embodiments, the linker "L" is a connector with a linear number of atoms other than hydrogen, ranging from 1 to 20. The connector "L" may contain, but is not limited to, functional groups such as ether, amide, alkane, alkene, alkyne, ketone, hydroxyl, carboxylic acid, thioether, sulfoxide and sulfone. The linker may contain aromatic, heteroaromatic, cyclic, bicyclic and tricyclic moieties. A halogen substitution may be included in the linker, such as Cl, F, Br, and I. In the case of fluorine substitution, one or more fluorines may be included.

[0025] In certain embodiments, VLM is a trans-3-hydroxyproline derivative wherein both the nitrogen and the carboxylic acid in trans-3-hydroxyproline are functionalized as amides.

[0026] In certain embodiments, CLM is a piperidine-2,6-dione derivative, wherein the piperidine-2,6-dione may be substituted at the 3-position and the 3-substitution may be a bicyclic heteroaromatic compound with a C-N bond or a C bond -WITH. Examples of CLMs include, but are not limited to, pomalidomide, lenalidomide and thalidomide and their derivatives.

[0027] In a further aspect, the disclosure provides therapeutic compositions containing an effective amount of a compound as described herein, or a salt form thereof, and a pharmaceutically acceptable carrier. Therapeutic compositions modulate degradation and/or inhibition of a protein in a patient or subject, for example, an animal such as a human, and can be used to treat or improve disease states or conditions that are modulated by the degraded/inhibited protein. In certain embodiments, the therapeutic compositions described herein can be used to cause the degradation of proteins of interest to treat or alleviate a disease, such as cancer (such as pancreatic cancer, colon cancer, colorectal cancer, lung cancer, or non-small cell lung cancer) . In another aspect, the present invention provides a method for ubiquitination/degradation of a target protein in a cell. In certain embodiments, the method comprises administering a bifunctional compound as described herein comprising ILM and PTM, PTM and VLM, or PTM and CLM, or PTM and MLM, preferably linked through a linker moiety as otherwise described herein, wherein VLM /ILM/CLM/MLM is linked to PTM through a linker to target a protein that binds to PTM for degradation. Likewise, a PTM may be linked to a VLM, or a CLM, or an MLM, or an ILM via a linker to target a protein or polypeptide for degradation. Degradation of the target protein will occur when the target protein is near the E3 ubiquitin ligase, resulting in degradation/inhibition of the effects of the target protein and control of protein levels. The control of protein levels provided by the present invention provides treatment for a disease state or pathological condition that is modulated by a target protein by reducing the level of that protein in the patient's cells.

[0028] In yet another aspect, the description provides methods of treating or alleviating a disease, pathology, or symptom thereof in a subject or patient, e.g., an animal such as a human, comprising administering to the subject in need thereof a composition containing an effective amount, e.g., a therapeutically effective amount an amount of a compound as described herein, or a salt form thereof, and a pharmaceutically acceptable carrier, wherein the composition is effective for treating or alleviating a disease or disorder or a symptom thereof in a subject.

[0029] In another aspect, the description provides methods for identifying the effects of degradation of proteins of interest in a biological system using compounds of the present invention.

[0030] The foregoing general scope is given by way of example only and is not intended to limit the scope of the present invention and the appended claims. Additional objects and advantages associated with the compositions, methods and processes of the present invention will be appreciated by one skilled in the art in light of the present claims, description and examples. For example, various aspects and embodiments of the invention can be used in numerous combinations, all of which are expressly suggested herein. These additional aspects and embodiments are clearly included within the scope of the present invention. Publications and other materials used herein to highlight the prior art of the invention and, in specific cases, to provide additional details regarding practice, are incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The accompanying figures, which are included in and constitute a part of the specification, illustrate several embodiments of the present invention and, together with the description, serve to explain the principles of the invention. The figures are intended to illustrate an embodiment of the invention only and should not be construed as limiting the invention. Additional objects, features and advantages of the invention will become apparent from the following detailed description taken in conjunction with the accompanying figures showing illustrative embodiments of the invention in which:

[0032] Figures 1A and 1B. Illustration of the general principle of the PROTAC function. (A) Typical PROTACs contain a protein targeting moiety (PTM; dark box), a ubiquitin ligase binding moiety (ULM; lightly shaded triangle), and an optional linker moiety (L; black line) connecting or linking the PTM to the ULM. (B) Illustrates the functional use of PROTAC as described herein. Briefly, ULM recognizes and binds to a specific E3 ubiquitin ligase, and PTM binds and recruits the target protein, bringing it closer to the E3 ubiquitin ligase. Typically, E3 ubiquitin ligase forms a complex with a protein conjugated to E2 ubiquitin and, either alone or through the E2 protein, catalyzes the addition of ubiquitin (dark circles) to a lysine on the target protein via an isopeptide bond. The polyubiquitinated protein (far right) is then degraded by the cell's proteasome machinery.

[0033] Figures 2A and 2B. Figure 2A is a Western blot showing a highly efficient degrader, typical compound 399. Figure 2B is a Western blot showing a less efficient degrader, typical compound 432. Both compounds covalently modify KRas ^G12C , as indicated by gel shift.

DETAILED DESCRIPTION OF THE INVENTION

[0034] The following is a detailed description provided to assist those skilled in the art in practicing the present invention. Modifications and changes may be made to the embodiments described herein by those skilled in the art without departing from the spirit and scope of the present invention. All publications, patent applications, patents, figures and other references mentioned herein are incorporated by reference in their entirety.

[0035] Compositions and methods have now been described that relate to the surprising and unexpected discovery that E3 ubiquitin ligase protein (e.g., inhibitors of apoptosis proteins (IAPs), Von Hippel-Lindau E3 ubiquitin ligase (VHL), cereblon E3 ubiquitin ligase, or E3 ubiquitin ligase mouse double minute homologue 2 (MDM2)) ubiquitinates the target protein once it and the target protein are placed in close proximity using a bifunctional or chimeric construct that links the E3 ubiquitin ligase protein and the target protein. Accordingly, the present invention provides such compounds and compositions comprising an E3 ubiquitin ligase binding moiety ("ULM") linked to a target protein binding moiety ("PTM"), resulting in ubiquitination of the selected target protein, resulting in degradation of the target protein. protein by the proteasome (see Figure 1). The present invention also provides a library of compositions and their use.

[0036] In certain aspects, the present invention provides compounds that contain a ligand, such as a low molecular weight ligand (i.e., having a molecular weight below 2000, 1000, 500, or 200 Daltons) that is capable of binding to a ubiquitin ligase such as IAP, VHL, MDM2 or Cereblon. The compounds also contain a moiety that is capable of binding to a target protein such that the target protein is placed adjacent to a ubiquitin ligase to cause degradation (and/or inhibition) of the protein. A small molecule may mean, in addition to the above, that the molecule is not a peptidyl, that is, it is generally not considered a peptide, for example, contains less than 4, 3 or 2 amino acids. As used herein, the PTM, ULM, or PROTAC molecule may be a small molecule.

[0037] Unless otherwise specified, all technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description is intended to describe specific embodiments and is not intended to limit the invention.

[0038] Where a range of values is proposed, it should be understood that each intermediate value, up to the tenth unit of the lower limit, unless the context clearly indicates otherwise (for example, in the case of a group containing multiple carbon atoms, in which case the number of each atom carbon is within the proposed range), between the upper and lower limits of this range and any other specified or intermediate value within this specified range is covered within the scope of the invention. The upper and lower limits of these smaller ranges may be independently included within the smaller ranges and are also covered within the scope of this invention, subject to any specifically excluded limit within said range. Where a specified range includes one or both limits, ranges excluding either or both of these included limits are also included in the invention.

[0039] The following terms are used to describe the present invention. Where a term is not specifically defined herein, the term is given the meaning recognized in the art by those of ordinary skill in the art when using the term in the context of its use in describing the present invention.

[0040] The singular, as used herein, is used to refer to one or more than one (ie, at least one) grammatical entity, unless the context clearly indicates otherwise. By way of example, "element" means one element or more than one element.

[0041] The phrase “and/or,” as used herein, is to be understood to mean “either or both” of the elements so combined, that is, elements that are present together in some cases and separately in other cases. Multiple elements listed by “and/or” are to be construed identically, that is, “one or more” elements so combined. Optionally, other elements other than those specifically identified by the statement “and/or” may be present, whether or not related to those specifically identified elements. Thus, as a non-limiting example, a reference to “A and/or B”, when used in conjunction with non-limiting language such as “comprising”, may refer in one embodiment to A only (not necessarily 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); and so on.

[0042] As used herein, the conjunction “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” should be interpreted as inclusive, i.e. including at least one, but also more than one, of a number or list of elements, and optionally additional elements not included in the list. Only terms clearly indicating the contrary, such as “only one of” or “exactly one of” or, when used in a claim, “consisting of”, will refer to the inclusion of exactly one element from a number or list of elements. In general, the term “or” as used herein should be interpreted only as indicating exclusive alternatives (i.e., “one or the other, but not both”) when preceded by exclusivity terms such as “either,” “ one of", "only one of" or "exactly one of".

[0043] In the claims, as well as in the above description, all transitional phrases such as “comprising”, “comprising”, “bearing”, “having”, “containing”, “involving”, “holding”, “consisting of” ", and so on. should be understood as unlimited, that is, meaning inclusion, but not without limitation. Only the transitional phrases “consisting of” and “consisting essentially of” must be limiting or semi-limiting transitional phrases, respectively, as specified in the USPTO Manual of Patent Examination Procedures, Section 2111.03

[0044] As used herein, the phrase "at least one" with respect to a list of one or more elements is to be understood to mean at least one element selected from any one or more elements in the list of elements, but not necessarily including at least one at least one of each element specifically listed in the list of elements, and without excluding any combination of elements in the list of elements. This definition also allows that elements may not necessarily be present beyond those specifically identified in the list of elements to which the phrase “at least one” refers, whether or not they are related to those specifically identified elements. Thus, by way of 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" ) may refer, in one embodiment, to at least one, optionally including more than one A, without the presence of B (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one B, without the presence of A (and optionally including elements other than A); in yet another embodiment, at least one, optionally including more than one A, and at least one, optionally including more than one B (and optionally including other elements).4 and the like.

[0045] It should also be understood that in some methods described 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 listed, unless the context indicates otherwise.

[0046] The terms "co-administration" and "co-administered" or "combination therapy" refer to both simultaneous administration (administration of two or more therapeutic agents at the same time) and time-varying administration (administration of one or more therapeutic agents at the same time). the same time, different from the time of administration of the additional therapeutic agent or agents) as long as the therapeutic agents are present in the patient's body to some extent, preferably in effective amounts, at the same time. In some preferred aspects, one or more compounds of the invention described herein are co-administered in combination with at least one additional bioactive agent, especially including an antineoplastic agent, such as chemotherapy or biological therapy targeting epidermal growth factor receptors (EGF receptors). for example, epidermal growth factor receptor inhibitors such as at least one of gefitinib, erlotinib, neratinib, lapatinib, cetuximab, vandetanib, necitumamab, osimertinib, or combinations thereof). In particularly preferred aspects, coadministration of the compounds results in synergistic activity and/or therapy, including anticancer activity.

[0047] As used herein, the term "compound", unless otherwise indicated, refers to any specific chemical compound disclosed herein and includes tautomers, regioisomers, geometric isomers and, where applicable, stereoisomers, including optical isomers thereof (enantiomers) and other stereoisomers (diastereomers), as well as pharmaceutically acceptable salts and derivatives, including prodrugs and/or deuterated forms thereof, where applicable, in the context. Deuterated small molecules are considered, in which one or more hydrogen atoms contained in the drug molecule are replaced by deuterium.

[0048] As used herein, the term compound generally refers to a single compound, but may also include other compounds such as stereoisomers, regioisomers and/or optical isomers (including racemic mixtures), as well as specific enantiomers or enantiomerically enriched mixtures of the disclosed compounds. The term also refers in context to prodrug forms of compounds that have been modified to facilitate administration and delivery of the compounds to the site of activity. It should be noted that in the description of the proposed compounds, among other things, numerous substituents and variables associated therewith are described. Those of ordinary skill in the art will appreciate that the molecules described herein are stable compounds, as generally described below. When a bond is shown, both the double bond and the single bond are presented or understood in the context of the compound shown and the well-known rules of valence interactions.

[0049] The term "ubiquitin ligase" refers to a family of proteins that facilitate the transfer of ubiquitin to a specific substrate protein, targeting that substrate protein for degradation. For example, IAP is an E3 ubiquitin ligase protein that, alone or in combination with an E2 ubiquitin-conjugated enzyme, causes the attachment of ubiquitin to a lysine on a target protein and subsequently targets specific protein substrates for degradation by the proteasome. Thus, E3 ubiquitin ligase alone or in complex with an E2 ubiquitin-conjugated enzyme is responsible for the transfer of ubiquitin to target proteins. Typically, a ubiquitin ligase is involved in polyubiquitination, so that a second ubiquitin is added to the first; the third is attached to the second and so on. Polyubiquitination marks proteins for degradation by the proteasome. However, there are some ubiquitination events that are limited to monoubiquitination, in which only one ubiquitin is added by a ubiquitin ligase to a substrate molecule. Monoubiquitinated proteins are not targeted to the proteasome for degradation, but instead can be altered in their cellular location or function, for example by binding to other proteins that have domains capable of binding ubiquitin. Further complicating matters is that different ubiquitin lysines can be targeted by E3 to form chains. The most common lysine is Lys48 in the ubiquitin chain. It is a lysine used to produce polyubiquitin, which is recognized by the proteasome.

[0050] The term “patient” or “subject” is used throughout this specification to describe an animal, preferably a human or domestic animal, to be treated, including prophylactic treatment, with the compositions of the present invention. For the treatment of those infections, pathological conditions or disease states that are specific to a particular animal, such as a human patient, wherein the term patient refers to that particular animal, including a domestic animal such as a dog or cat, or a farm animal such as a horse , cow, sheep, etc. In general, as used herein, the term patient refers to a human patient unless otherwise stated or implied by the context of use of the term.

[0051] The term “effective” is used to describe the amount of a compound, composition, or component that, when applied in the context of its intended use, produces the desired result. The term effective includes all other terms, such as effective amount or effective concentration, that are otherwise described or used in this application.

[0052] Compounds and compositions

[0053] In one aspect, the disclosure provides compounds comprising an E3 ubiquitin ligase binding fragment ("ULM"), which is an E3 ubiquitin ligase IAP binding fragment ("ILM"), a cereblon E3 ubiquitin ligase binding fragment ("CLM"), a fragment von Hippel-Lindau (VHL) E3 ubiquitin ligase binding fragment (VLM), and/or mouse double minute homolog 2 (MDM2) E3 ubiquitin ligase binding fragment (MLM). In an exemplary embodiment, the ULM is linked to a target protein binding moiety (PTM) via a chemical linker (L) according to the structure:

(A) PTM-L-ULM

where L represents a bond or chemical linker group, ULM is an E3 ubiquitin ligase binding moiety, and PTM is a target protein binding moiety. The number and/or relative positions of moieties in the compounds illustrated herein are offered by way of example only. As will be appreciated by one skilled in the art, the compounds described herein can be synthesized with any desired number and/or relative position of the corresponding functional moieties.

[0054] The terms ULM, ILM, VLM, MLM and CLM are used in their inclusive sense unless the context indicates otherwise. For example, the term ULM includes all ULMs, including those that bind IAP (ie, ILM), MDM2 (ie, MLM), Cereblon (ie, CLM) and VHL (ie, VLM). Additionally, the term ILM includes all possible E3 ubiquitin ligase IAP binding fragments, the term MLM includes all possible E3 ubiquitin ligase MDM2 binding fragments, the term VLM includes all possible VHL binding fragments, and the term CLM includes all cereblon binding fragments.

[0055] In another aspect, the present invention provides bifunctional or multifunctional compounds (eg, PROTACs) useful for regulating protein activity by inducing degradation of a target protein. In certain embodiments, the compound comprises an ILM, or a VLM, or a CLM, or an MLM linked, for example linked covalently, directly or indirectly, to a moiety that binds the target protein (i.e., a protein targeting moiety or "PTM") . In certain embodiments, the ILM/VLM/CLM/MLM and the PTM are connected or linked through a chemical linker (L). ILM binds the E3 ubiquitin ligase IAP, VLM binds VHL, CLM binds the E3 ubiquitin ligase cereblon and MLM binds the E3 ubiquitin ligase MDM2, and PTM recognizes the target protein and the interaction of the corresponding fragments with their targets facilitates the degradation of the target protein by placing the target protein in close proximity to the protein ubiquitin ligases. A typical bifunctional compound can be depicted as:

(B) RTM-ILM

(C) RTM-CLM

(D) RTM-VLM

(E) RTM-MLM

[0056] In some embodiments, the bifunctional compound further comprises a chemical linker (“L”). For example, a bifunctional compound can be represented as:

(F) RTM-L-ILM

(G) RTM-L-CLM

(H) RTM-L-VLM

(I) RTM-L-MLM

where PTM is a protein/polypeptide targeting fragment, L is a chemical linker, ILM is an E3 ubiquitin ligase IAP binding fragment, CLM is a cereblon E3 ubiquitin ligase binding fragment, VLM is a VHL binding fragment, and MLM is E3 ubiquitin ligase MDM2 binding fragment.

[0057] In certain embodiments, a ULM (e.g., ILM, CLM, VLM, or MLM) is active in or binds to an E3 ubiquitin ligase (e.g., E3 ubiquitin ligase IAP, cereblon E3 ubiquitin ligase, VHL, or E3 ubiquitin ligase MDM2) with an IC ₅₀ of less than about 200 µM. can be determined by any method known in the art, such as fluorescent polarization analysis.

[0058] In certain additional embodiments, the bifunctional compounds described herein exhibit activity with an IC ₅₀ of less than about 100, 50, 10, 1, 0.5, 0.1, 0.05, 0.01, 0.005 ,0.001 mM, or less than about 100, 50,10, 1,0.5, 0.1, 0.05,0.01,0.005,0.001 µM, or less than about 100, 50, 10, 1 , 0.5, 0.1, 0.05, 0.01, 0.005, 0.001 nM, or less than about 100, 50, 10, 1, 0.5, 0.1, 0.05, 0.01 , 0.005, 0.001 pM.

[0059] In certain embodiments, the compounds described herein contain multiple PTMs (targeted to the same or different protein targets), multiple ULMs, one or more ULMs (i.e., moieties that specifically bind to multiple/ different E3 ubiquitin ligases, for example VHL, IAP, Cereblon and/or MDM2) or a combination thereof. In any of the aspects or embodiments described herein, PTM and UTM (eg, ILM, VTM, CEM and/or MLM) can be linked directly, or through one or more chemical linkers, or a combination thereof. In further embodiments, where a compound has multiple ULMs, the ULMs may be for the same E3 ubiquitin ligase, or each respective ULM may specifically bind to a different E3 ubiquitin ligase. In other further embodiments, where a compound has multiple PTMs, the PTMs may bind to the same target protein, or each respective PTM may specifically bind to a different target protein.

[0060] In certain embodiments where a connection contains multiple ULMs, the ULMs are identical. In additional embodiments, a compound containing multiple ULMs (eg, ULM, ULM', etc.), at least one PTM is linked to the ULM directly, or through a chemical linker (L), or both. In certain further embodiments, a connection comprising multiple ULMs further comprises multiple PTMs. In other further embodiments, the PTMs are the same or optionally different. In other embodiments, where the PTMs are different, the respective PTMs may bind to the same target protein or specifically bind to a different target protein.

[0061] In certain embodiments, a connection may comprise a plurality of ULMs and/or a plurality of ULM's. In further embodiments, the compound comprising at least two different ULMs, multiple ULMs, and/or multiple ULMs', further comprises at least one PTM linked to the ULM or ULM' directly, or via a chemical linker, or both . In any of the embodiments described herein, a connection comprising at least two different ULMs may further comprise multiple PTMs. In other further embodiments, the PTMs are the same or optionally different. In other further embodiments, where the PTMs are different, the respective PTMs may bind to the same target protein or specifically bind to a different target protein. In other further embodiments, the PTM is inherently a ULM (or ULM'), such as ILM, VLM, CLM, MLM, ILM', VLM', CLM' and/or MLM'.

[0062] In additional embodiments, the disclosure provides compounds as described herein, including enantiomers, diastereomers, solvates, and polymorphs thereof, including pharmaceutically acceptable salt forms thereof, eg, acid and base salt forms.

[0063] Typical ILMs

[0064] Tetrapeptide fragments of AVPI

[0065] In any of the compounds described herein, the ILM may contain a tetrapeptide fragment alanine-valine-proline-isoleucine (AVPI) or a non-natural mimetic thereof. In certain embodiments, the ELM is selected from the group consisting of chemical structures provided as formulas (I), (II), (III), (IV) and (V):

Where:

R ¹ for formulas (I), (II), (III), (IV) and (V) is selected from H or alkyl;

R ² for formulas (I), (II), (III), (IV) and (V) is selected from H or alkyl;

R ³ for formulas (I), (II), (III), (IV) and (V) is selected from H, alkyl, cycloalkyl and heterocycloalkyl;

R ⁵ and R ⁶ for formulas (I), (II), (III), (IV) and (V) are independently selected from H, alkyl, cycloalkyl, heterocycloalkyl, or more preferably, R ⁵ and R ⁶ are taken together for formulas ( I), (II), (III), (IV) and (V) to form a pyrrolidine or piperidine ring optionally further fused with 1-2 cycloalkyl, heterocycloalkyl, aryl or heteroaryl rings, each of which can then be further fused with the other a cycloalkyl, heterocycloalkyl, aryl or heteroaryl ring;

R ³ and R ⁵ for formulas (I), (II), (III), (IV) and (V) taken together may form a 5-8 membered ring, optionally further fused with 1-2 cycloalkyl, heterocycloalkyl, aryl or heteroaryl;

R ⁷ for formulas (I), (II), (III), (IV) and (V) is selected from cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, aryl-C(O)-R ⁴ , arylalkyl, heteroaryl, heteroaryl -C(O)-R ⁴ , heteroaryl-R ⁴ , heteroaryl-naphthalene, heteroarylalkyl or -C(O)NH-R ⁴ , each of which is further optionally substituted with 1-3 substituents selected from halogen, alkyl, haloalkyl, hydroxyl, alkoxy, cyano, (hetero)cycloalkyl, aryl, (hetero)aryl, -C(O)NH-R ⁴ or C(O)-R ⁴ ; And

R ⁴ is selected from alkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, aryl (eg, bicyclic aryl), arylalkyl, heteroaryl (eg, bicyclic heteroaryl), heteroarylalkyl, further optionally substituted with 1-3 substituents as described above.

[0066] As shown above, P1, P2, P3 and P4 of formula (II) are in connection with A, V, P and I, respectively, of the tetrapeptide fragment of AVPI or a non-natural mimetic thereof. Likewise, each of formulas (I) and (III) to (V) has portions associated with A, V, P and I of the tetrapeptide fragment of AVPI or a non-natural mimetic thereof.

[0067] In any of the compounds described herein, the ILM may have a structure of formula (VI), which is a derivative of the IAP antagonists described in WO publication No. 008/014236, or a non-natural mimetic thereof:

Where:

R ₁ of formula (VI) is independently selected from H, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkenyl, C ₁ -C ₄ alkynyl or C ₃ -C ₁₀ cycloalkyl, which are unsubstituted or substituted;

R ₂ of formula (VI) is independently selected from H, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkenyl, C ₁ -C ₄ alkynyl or C ₃ -C ₁₀ cycloalkyl, which are unsubstituted or substituted;

R ₃ of formula (VI) is independently selected from H, -CF ₃ , -C ₂ H ₅ , C ₁ -C ₄ -alkyl, C ₁ -C ₄ -alkenyl, C ₁ -C ₄ -alkynyl, - CH ₂ -Z , or any R ₂ and R ₃ together form a heterocyclic ring;

each Z of formula (VI) is independently selected from H, -OH, F, Cl, -CH ₃ , -CF ₃ , -CH ₂ Cl, -CH ₂ F or -CH ₂ OH;

R ₄ of formula (VI) is independently selected from C ₁ -C ₁₆ straight or branched alkyl, C ₁ -C ₁₆ alkenyl, C ₁ -C ₁₆ alkynyl, C ₃ -C ₁₀ cycloalkyl. -(CH ₂ ) _0-6 -Z ₁ , -(CH ₂ ) _0-6 -aryl and -(CH ₂ ) _0-6 -het, where alkyl, cycloalkyl and phenyl are unsubstituted or substituted;

R ₅ of formula (VI) is independently selected from H, C _1-10 -alkyl, aryl, phenyl, C _3-7 -cycloalkyl, -(CH ₂ ) _0-6 -C _3-7 -cycloalkyl, -C _1-10 -alkyl-aryl, -(CH ₂ ) _0-6 -C _3-7 -cycloalkyl-(CH ₂ ) _0-6 -phenyl, -(CH ₂ ) _0-4 -CH[(CH ₂ ) _1-4 - phenyl] ₂ , indanyl, -C(O)-C _1-10 -alkyl, -C(O)-(CH ₂ ) _1-6 -C _3-7 -cycloalkyl, -C(O)-(CH ₂ ) _0-6 -phenyl, - (CH ₂ ) _0-6 -C(O)-phenyl, -(CH ₂ ) _0-6 -het, -C(O)-(CH ₂ ) _1-6 -het, or R ₅ is selected from an amino acid residue where alkyl, cycloalkyl, phenyl, and aryl are unsubstituted or substituted;

Z ₁ of formula (VI) is independently selected from -N(R ₁₀ )-C(O)-C _1-10 -alkyl, -N(R ₁₀ )-C(O)-(CH ₂ ) _0-6 -C _{3 -7} -cycloalkyl, -N(R ₁₀ )-C(O)-(CH ₂ ) _0-6 -phenyl, -N(R ₁₀ )-C(O)(CH ₂ ) _1-6 -het, -C (O)-N(R ₁₁ )(R ₁₂ ), -C(O)-O-C _1-10 -alkyl, -C(O)-O-(CH ₂ ) _1-6 -C _3-7 - cycloalkyl, -C(O)-O-(CH ₂ ) _0-6 -phenyl, -C(O)-O-(CH ₂ ) _1-6 -het, -OC(O)-C _1-10 -alkyl , -O-C(O)-(CH ₂ ) _1-6 -C _3-7 -cycloalkyl, -O-C(O)-(CH ₂ ) _0-6 -phenyl, - OC(O)-(CH ₂ ) _1-6 -het, where alkyl, cycloalkyl and phenyl are unsubstituted or substituted;

het of formula (VI) is independently selected from a 5-7 membered heterocyclic ring containing 1-4 heteroatoms selected from N, O and S, or an 8-12 membered fused ring system containing at least one 5-7 membered heterocyclic ring, containing 1, 2 or 3 heteroatoms selected from N, O and S, the heterocyclic ring or fused ring system of which is unsubstituted or substituted at a carbon or nitrogen atom;

R ₁₀ of formula (VI) is selected from H, -CH ₃ , -CF ₃ , -CH ₂ OH, or -CH ₂ Cl;

R ₁₁ and R ₁₂ of formula (VI) are independently selected from H, C _1-4 -alkyl, C _3-7 -cycloalkyl, -(CH ₂ ) _1-6 -C _3-7 -cycloalkyl, (CH ₂ ) _{0- 6} -phenyl, where alkyl, cycloalkyl and phenyl are unsubstituted or substituted; or R ₁₁ and R ₁₂ together with nitrogen form het, and

U of formula (VI) is, independently, as shown in formula (VII):

Where:

each n of formula (VII) is independently selected from 0-5;

X of formula (VII) is selected from the group -CH and N;

R _a and R _b , formula (VII) are independently selected from the group of O, S or N, or C _0-8 -alkyl atoms, wherein one or more carbon atoms in the alkyl chain are optionally replaced by a heteroatom selected from O, S or N, and, wherein each alkyl is, independently, either unsubstituted or substituted;

R _d of formula (VII) is selected from the group Re-Q-(R _f ) _p (R _g ) _q , and Ar ₁ -D-Ar ₂ ;

R _c of formula (VII) is selected from the group H, or any R _c and R _d together form cycloalkyl or het; where, if R _c and R _d form a cycloalkyl or net, R ₅ is attached to the formed ring to a C or N atom;

p and q of formula (VII) are independently selected from 0 or 1;

R _e of formula (VII) is selected from the group C _1-8 -alkyl and alkylidene, and each R _e is either unsubstituted or substituted; Q is selected from the group N, O, S, S(O) and S(O) ₂ ;

Ar ₁ and Ar ₂ of Formula (VII) are independently selected from the group of substituted or unsubstituted aryl and het;

R _f and R _g of formula (VII) are independently selected from H, -C _1-10 -alkyl, C _1-10 -alkylaryl, -OH, -O-C _1-10 -alkyl, - (CH ₂ ) _0-6 -C _3-7 -cycloalkyl, -O-(CH ₂ ) _0-6 -aryl, phenyl, aryl, phenyl-phenyl, -(CH ₂ ) _1-6 -het, -O-(CH ₂ ) _1-6 -het, -OR ₁₃ , -C(O)-R ₁₃ , -C(O)-N(R ₁₃ )(R ₁₄ ), -N(R ₁₃ )(R ₁₄ ), -SR ₁₃ , -S( O)-R ₁₃ , -S(O) ₂ -R ₁₃ , -S(O) ₂ -NR ₁₃ R ₁₄ , -NR ₁₃ -S(O) ₂ -R ₁₄ , -SC _t-10 -alkyl, aryl -C _1-4 -alkyl or het-C _1-4 -alkyl, where alkyl, cycloalkyl, het and aryl are unsubstituted or substituted, -SO ₂ -C _1-2 -alkyl, -SO ₂ -C _1-2 - alkylphenyl, -OC _1-4 -alkyl, or any R _g and R _f together form a ring selected from het or aryl;

D of Formula (VII) is selected from the group -CO-, -C(O)-C _1-7 -alkylene or arylene, -CF ₂ -, -O-, -S(O) _r where r is 0-2, 1,3-dioxalane, or C _1-7 -alkyl-OH; where alkyl, alkylene or arylene is unsubstituted or substituted with one or more halogens, OH, -O-C1-6-alkyl, -SC _1-6 -alkyl or -CF ₃ ; or each D is independently selected from N(R _h );

R _h is selected from the group H, unsubstituted or substituted C _1-7 -alkyl, aryl, unsubstituted or substituted -O-(C _1-7 -cycloalkyl), -C(O)-C _1-10 -alkyl, -C( O)-C _1-10 -alkyl-aryl, -COC _01-10 -alkyl, -C-O-C _0-10 -alkyl-aryl, -SO ₂ -C _1-10 -alkyl or -SO ₂ -( C _0-10 -alkylaryl);

R ₆ , R ₇ , R ₈ and R ₉ of formula (VII) are independently selected from the group H, -C _1-10 -alkyl, -C _1-10 -alkoxy, aryl-C _1-10 -alkoxy, -OH, - O-C _1-10 -alkyl, -(CH ₂ ) _0-6 -C _3-7 -cycloalkyl, -O-(CH ₂ ) _0-6 -aryl, phenyl, -(CH ₂ ) _1-6 -het , -O-(CH ₂ ) _1-6 -het, -OR ₁₃ , -C(O)-R ₁₃ , -C(O)-N(R ₁₃ )(R ₁₄ ), -N(R ₁₃ )( R ₁₄ ), -SR ₁₃ , -S(O)-R ₁₃ , -S(O) ₂ -R ₁₃ , -S(O) ₂ -NR ₁₃ R ₁₄ or -NR ₁₃ -S(O) ₂ -R ₁₄ ; wherein each alkyl, cycloalkyl and aryl is unsubstituted or substituted; and any R ₆ , R ₇ , R ₈ and R ₉ optionally together form a ring system;

R ₁₃ and R ₁₄ Formulas (VII) are independently selected from the group H, C _1-10 -alkyl, -(CH ₂ ) _0-6 -C _3-7 -cycloalkyl, -(CH ₂ ) _0-6 - (CH) _0-1 -(aryl) _1-2 , -C(O)-C _1-10 -alkyl, -C(O)-(CH ₂ ) _1-6 -C _3-7 -cycloalkyl, -C(O) -O-(CH ₂ ) _0-6 -aryl, - C(O)-(CH ₂ ) _0-6 -O-fluorenyl, -C(O)-NH-(CH ₂ ) _0-6 -aryl, - C(O)-(CH ₂ ) _0-6 -aryl, -C(O)-(CH ₂ ) _0-6 -het, - C(S)-C _1-10 -alkyl, -C(S)- (CH ₂ ) _1-6 -C _3-7 -cycloalkyl, -C(S)-O-(CH ₂ ) _0-6 -aryl, -C(S)-(CH ₂ ) _0-6 -O-fluorenyl , -C(S)-NH-(CH ₂ ) _0-6 -aryl, -C(S)-(CH ₂ ) _0-6 -aryl, or -C(S)-(CH ₂ ) _1-6 - het, where each alkyl, cycloalkyl, and aryl is unsubstituted or substituted: or any R ₁₃ and R ₁₄ together with a nitrogen atom form het;

wherein the alkyl substituents in R ₁₃ and R ₁₄ of Formula (VII) are unsubstituted or substituted and when substituted, are substituted with one or more substituents selected from C _1-10 -alkyl, halogen, OH, - O-C _1-6 -alkyl, -S-C _1-6 -alkyl and -CF ₃ ; and the substituted phenyl or aryl in R ₁₃ and R ₁₄ is substituted with one or more substituents selected from halogen, hydroxyl, C _1-4 alkyl, C _1-4 alkoxy, nitrogen, -CN, -O-C(O)- C _1-4 -alkyl, and -C(O)-O-C _1-4 -aryl; or a pharmaceutically acceptable salt or hydrate thereof.

[0068] In certain embodiments, the compound further comprises an independently selected second ILM attached to the ILM of Formula (VI), or a non-natural mimetic thereof, via at least one additional independently selected linker group. In one embodiment, the second ILM is a derivative of Formula (VI) or a non-natural mimetic thereof. In a certain embodiment, the at least one additional independently selected linker group comprises two additional independently selected linker groups that chemically link the ILM and the second ILM. In one embodiment, at least one additional linker group for the ILM of formula (VI) or a non-natural mimetic thereof chemically links groups selected from R ₄ and R ₅ . For example, an ILM of formula (VI) and a second ILM of formula (VI) or a non-natural mimetic thereof may be related as follows:

[0069] In certain embodiments of the ILM, at least one additional independently selected linker group L and the second ILM have a structure selected from the group consisting of:

which are derivatives of the IAP antagonists described in WO publication No. 2008/014236.

[0070] In any of the compounds described herein, the ILM may have the structure of formula (VIII), which is based on the LAP ligands described in Ndubaku, C, et al. Antagonism of c-IAP and XIAP proteins is required for effective induction of cell death by small molecule LAP antagonists, ACS Chem. Biol., 557-566, 4 (7) (2009) or its non-natural mimetic:

wherein each of A1 and A2 of formula (VIII) is independently selected from optionally substituted monocyclic fused rings, aryls and heteroaryls; and

R of formula (VIII) is selected from H or Me.

[0071] In a particular embodiment, the linker group L is attached to A1 of formula (VIII). In another embodiment, the linker group L is attached to A2 of formula (VIII).

[0072] In a particular embodiment, the ILM is selected from the group consisting of

[0073] In any of the compounds described herein, the ILM may have a structure of formula (IX), which is derived from the chemotypes specified in Mannhold, R., et al. IAP antagonists: promising candidates for cancer therapy, Drug Discov. Today, 15 (5-6), 210-9 (2010) or its non-natural mimetic:

where R ¹ is selected from alkyl, cycloalkyl and heterocycloalkyl and, most preferably, from isopropyl, tert-butyl, cyclohexyl and tetrahydropyranyl, and R ² of formula (IX) is selected from -OPh or H.

[0074] In any of the compounds described herein, the ILM may have a structure of formula (X), which is derived from the chemotypes specified in Mannhold, R., et al. IAP antagonists: promising candidates for cancer therapy, Drug Discov. Today, 15 (5-6), 210-9 (2010) or its non-natural mimetic:

Where:

R ¹ of formula (X) is selected from H, -CH ₂ OH, --CH ₂ CH ₂ OH, --CH ₂ NH ₂ , --CH ₂ CH ₂ NH ₂ ; X of formula (X) is selected from S or CH ₂ ;

R ² of formula (X) is selected from:

R ³ and R ⁴ of formula (X) are independently selected from H or Me

[0075] In any of the compounds described herein, the ILM may have a structure of formula (XI), which is derived from the chemotypes specified in Mannhold, R., et al. IAP antagonists: promising candidates for cancer therapy, Drug Discov. Today, 15 (5-6), 210-9 (2010) or its non-natural mimetic:

where R ¹ of formula (XI) is selected from H or Me, and R ² of formula (XI) is selected from H or

[0076] In any of the compounds described herein, the ILM may have the structure of formula (XII), which is derived from the chemotypes specified in Mannhold, R., et al. IAP antagonists: promising candidates for cancer therapy, Drug Discov. Today, 15 (5-6), 210-9 (2010) or its non-natural mimetic:

Where:

R ¹ of formula (XII) is selected from:

And

R ² of formula (XII) is selected from:

[0077] In any of the compounds described herein, the E3 ubiquitin ligase IAP binding moiety is selected from the group consisting of:

And

[0078] In any of the compounds described herein, the ILM may have the structure of formula (XIII), which is based on the ligands summarized in Ftygare, J.A., et al. Small-molecule pan-IAP antagonists: a patent review. Expert Gpin. Ther. Pat, 20(2), 251-67 (2010) or its non-natural mimetic:

Where:

Z of formula (XIII) is absent or O;

R ¹ of formula (XIII) is selected from:

R ¹⁰ V selected from H, alkyl, or aryl;

X is selected from CH ₂ and O; And

is a nitrogen-containing heteroaryl.

[0079] In any of the compounds described herein, the ILM may have the structure of formula (XIV), which is based on the IAP ligands summarized in Ftygare, J.A., et al. Small-molecule pan-IAP antagonists: a patent review, Expert Opin. Ther. Pat, 20(2), 251-67 (2010) or its non-natural mimetic:

Where:

Z of formula (XIV) is absent or O;

R ³ and R ⁴ of formula (XIV) are independently selected from H or Me;

R1 of formula (XIV) is selected from:

R ¹⁰ V selected from H, alkyl, or aryl;

X in selected from CH ₂ and O; And

V or is a nitrogen-containing heteroaryl.

[0080] In any of the connections described herein, the ILM is selected from the group consisting of:

And

which are derivatives of the ligands disclosed in US Patent Publication No. 2008/0269140 and US Patent No. 7244851.

[0081] In any of the compounds described herein, the ILM may have a structure of formula (XV), which was derived from the IAP ligand described in WO publication No. 2008/128171, or a non-natural mimetic thereof:

Where:

Z of formula (XV) is absent or O;

R ¹ of formula (XV) is selected from:

R ¹⁰ V selected from H, alkyl, or aryl;

X in selected from CH ₂ and O; And

V or is a nitrogen-containing heteroaryl; And

R ² of formula (XV) is selected from H, alkyl or acyl;

[0082] In a specific embodiment, the ILM has the following structure:

[0083] In any of the compounds described herein, the ILM may have a structure of formula (XVI), which is based on the IAP ligand described in WO publication No. 2006/069063, or a non-natural mimetic thereof:

Where:

R ² of formula (XVI) is selected from alkyl, cycloalkyl and heterocycloalkyl; more preferably isopropyl, tert-butyl, cyclohexyl and tetrahydropyranyl, most preferably cyclohexyl;

formula (XVI) is a 5- or 6-membered nitrogen-containing heteroaryl; more preferably a 5-membered nitrogen heteroaryl, and most preferably a thiazole; and Ar of formula (XVI) is aryl or heteroaryl.

[0084] In any of the compounds described herein, the ILM may have a structure of formula (XVII), which is based on the IAP ligands described in Cohen, F. et al., Antagonists of inhibitors of apoptosis proteins based on thiazole amide isosteres, Bioorg. Med. Chem. Lett., 20(7), 2229-33 (2010) or its non-natural mimetic:

Where:

R ¹ of formula (XVII) is selected from the group of halogen (for example, fluorine), cyano,

X of formula (XVII) is selected from the group O or CH ₂ .

[0085] In any of the compounds described herein, the ILM may have a structure of formula (XVIII), which is based on the IAP ligands described in Cohen, F. et al., Antagonists of inhibitors of apoptosis proteins based on thiazole amide isosteres, Bioorg. Med. Chem. Lett., 20(7), 2229-33 (2010) or its non-natural mimetic:

where R of formula (XVIII) is selected from alkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl or halogen (in variable substitution position).

[0086] In any of the compounds described herein, the ILM may have a structure of formula (XIX), which is based on the IAP ligands described in Cohen, F. et al., Antagonists of inhibitors of apoptosis proteins based on thiazole amide isosteres, Bioorg. Med. Chem. Lett., 20(7), 2229-33 (2010) or its non-natural mimetic:

Where is a 6-membered nitrogen-containing heteroaryl.

[0087] In a certain embodiment, the ILM of the composition is selected from the group consisting of:

[0088] In certain embodiments, the ILM of the composition is selected from the group consisting of:

And

[0089] In any of the compounds described herein, the ILM may have a structure of formula (XX), which is based on the IAP ligands described in WO publication No. 2007/101347, or a non-natural mimetic thereof:

where X of formula (XX) is selected from CH ₂ , O, NH or S.

[0090] In any of the compounds described herein, the ILM may have a structure of formula (XXI), which is based on the IAP ligands described in US Pat. No. 7,345,081 and US Pat. No. 7,419,975 or a non-natural mimetic thereof:

Where:

R ² of formula (XXI) is selected from:

R ⁵ of formula (XXI) is selected from And

W of formula (XXI) is selected from CH or N; And

R ⁶ in And are independently mono- or bicyclic fused aryl or heteroaryl.

[0091] In certain embodiments, the connection ILM is selected from the group consisting of:

[0092] In certain embodiments, the connection ILM is selected from the group consisting of:

And

which are described in WO Publication No. 2009/060292, US Patent No. 7517906, WO Publication No. 2008/134679, WO Publication No. 2007/130626, and WO Publication No. 2008/128121.

[0093] In any of the compounds described herein, the ILM may have the structure of formulas (XXII) or (XXIII), which are derivatives of the IAP ligands described in WO publication No. 2015/006524 and Perez HL, Discovery of potent heterodimeric antagonists of inhibitor of apoptosis proteins (IAPs) with sustained antitumor activity. J. Med. Chem. 58(3), 1556-62 (2015) or its non-natural mimetic:

or

Where:

R ¹ of formula (XXII) or (XXIII) is optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl, or optionally substituted aryl;

R ² of formula (XXII) or (XXIII) is optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl, or optionally substituted aryl; or alternatively, R ¹ and R ² of formula (XXII) or (XXIII) are independently optionally substituted thioalkyl, wherein the substituents attached to the S atom of the thioalkyl are optionally substituted alkyl, optionally substituted branched alkyl, optionally substituted heterocyclyl, - (CH ₂ ) _v COR ²⁰ , CH ₂ CHR ²¹ COR ²² or -CH ₂ R ²³ ; Where:

v is an integer from 1-3;

R ²⁰ and R ²² in -(CH ₂ ) _v COR ²⁰ and -CH ₂ R ²³ are independently selected from OH, NR ²⁴ R ²⁵ or OR ²⁶ ;

R ²¹ in -CH ₂ CHR ²¹ COR ² selected from the group NR ²⁴ R ²⁵ ;

R ²³ in -CH ₂ R ²³ is selected from optionally substituted aryl or optionally substituted heterocyclyl, where optional substituents include alkyl and halogen;

R ²⁴ in NR ²⁴ R ²⁵ is selected from hydrogen or optionally substituted alkyl;

R ²⁵ in NR ²⁴ R ²⁵ is selected from hydrogen, optionally substituted alkyl, optionally substituted branched alkyl, optionally substituted arylalkyl, optionally substituted heterocyclyl, -CH ₂ (OCH ₂ CH ₂ O) _m CH ₃ or a polyamine chain such as spermine or spermidine ;

R ²⁶ in OR ²⁶ is selected from optionally substituted alkyl, wherein the optional substituents are OH, halogen or NH ₂ ; And

m is an integer from 1-8;

R ³ and R ⁴ of formula (XXII) or (XXIII) are independently selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted arylalkoxy, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted heteroarylalkyl or optionally substituted heterocycloalkyl, where the substituents are alkyl, halogen or OH;

R ⁵ , R ⁶ , R ⁷ and R ⁸ of formula (XXII) or (XXIII) are independently selected from hydrogen, optionally substituted alkyl or optionally substituted cycloalkyl; And

X is selected from a bond or chemical linker group, and/or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[0094] In certain embodiments, X is a bond or is selected from the group consisting of:

where "*" represents the attachment point of a PTM, L, or ULM, such as an ILM.

[0095] In any of the compounds described herein, the ILM may have the structure of formulas (XXIV) or (XXVI), which are derivatives of the IAP ligands described in WO publication No. 2015/006524 and Perez HL, Discovery of potent heterodimeric antagonists of inhibitor of apoptosis proteins (IAPs) with sustained antitumor activity. J. Med Chem. 58(3), 1556-62 (2015) or its non-natural mimetic, and the chemical linker of the linker group L is as shown:

or

Where:

R ¹ of formula (XXIV), (XXV) or (XXVI) is selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl or optionally substituted aryl;

R ² of formula (XXIV), (XXV) or (XXVI) is selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl or optionally substituted aryl;

or alternatively,

R ¹ and R ² of formula (XXIV), (XXV) or (XXVI) are independently selected from optionally substituted thioalkyl, wherein the substituents attached to the S atom of the thioalkyl are optionally substituted alkyl, optionally substituted branched alkyl, optionally substituted heterocyclyl, -( CH ₂ ) _v COR ²⁰ , -CH ₂ CHR ²¹ COR ²² or -CH ₂ R ²³ , where:

v is an integer from 1-3;

R ²⁰ and R ²² in -(CH ₂ ) _v COR ²⁰ and -CH ₂ R ²³ are independently selected from OH, NR ²⁴ R ²⁵ or OR ²⁶ ;

R ²¹ in -CH ₂ CHR ²¹ COR ² selected from NR ²⁴ R ²⁵ ;

R ²³ in -CH ₂ R ²³ is selected from optionally substituted aryl or optionally substituted heterocyclyl, where optional substituents include alkyl and halogen;

R ²⁴ in NR ²⁴ R ²⁵ is selected from hydrogen or optionally substituted alkyl;

R ²⁵ in NR ²⁴ R ²⁵ is selected from hydrogen, optionally substituted alkyl, optionally substituted branched alkyl, optionally substituted arylalkyl, optionally substituted heterocyclyl, -CH ₂ (OCH ₂ CH ₂ O) _m CH ₃ or a polyamine chain such as spermine or spermidine ;

R ²⁶ in OR ²⁶ is selected from optionally substituted alkyl, wherein the optional substituents are OH, halogen or NH ₂ ; And

m is an integer from 1-8;

R ³ and R ⁴ of formula (XXIV), (XXV) or (XXVI) are independently optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted arylalkoxy, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted heteroarylalkyl or optionally substituted heterocycloalkyl, where the substituents are alkyl, halogen or OH;

R ⁵ , R ⁶ , R ⁷ and R ⁸ of formula (XXIV), (XXV) or (XXVI) are independently hydrogen, optionally substituted alkyl or optionally substituted cycloalkyl; and/or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[0096] In a specific embodiment of the ILM in accordance with formulas (XXII) to (XXVI):

R ⁷ and R ⁸ are selected from H or Me;

R ⁵ and R ⁶ are selected from the group consisting of:

R ³ and R ⁴ are selected from the group containing:

[0097] In any of the compounds described herein, the ILM may have the structure of formulas (XXVII) or (XXVIII), which are derivatives of the IAP ligands described in WO publication No. 2014/055461 and Kim, KS, Discovery of tetrahydroisoquinoline-based bivalent heterodimeric IAP antagonists. Bioorg. Med. Chem. Lett. 24(21), 5022-9 (2014) or its non-natural mimetic:

And

Where:

R ³⁵ represents 1-2 substituents selected from alkyl, halogen, alkoxy, cyano and haloalkoxy;

R ¹ of formula (XXVII) and (XXVIII) is selected from H or optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl or optionally substituted aryl;

R ² of formula (XXVII) and (XXVIII) is selected from H or optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl or optionally substituted aryl; or alternatively,

R ¹ and R ² of formula (XXVII) and (XXVIII) are independently selected from optionally substituted thioalkyl -CR ⁶⁰ R ⁶¹ SR ⁷⁰ , where R ⁶⁰ and R ⁶¹ are selected from H or methyl, and R ⁷⁰ is selected from optionally substituted alkyl, optionally substituted branched alkyl, optionally substituted heterocyclyl, -(CH ₂ ) _v COR ²⁰ , -CH ₂ CHR ²¹ COR ²² or -CH ₂ R ²³ , where:

v is an integer from 1-3;

R ²⁰ and R ²² in -(CH ₂ ) _v COR ²⁰ and -CH ₂ CHR ²¹ COR ²² are independently selected from OH, NR ²⁴ R ²⁵ or OR ²⁶ ;

R ²¹ in -CH ₂ CHR ²¹ COR ²² selected from NR ²⁴ R ²⁵ ;

R ²³ in -CH ₂ R ²³ is selected from optionally substituted aryl or optionally substituted heterocyclyl, where optional substituents include alkyl and halogen;

R ²⁴ in NR ²⁴ R ²⁵ is selected from hydrogen or optionally substituted alkyl;

R ²⁵ in NR ²⁴ R ²⁵ is selected from hydrogen, optionally substituted alkyl, optionally substituted branched alkyl, optionally substituted arylalkyl, optionally substituted heterocyclyl, -CH ₂ CH ₂ (OCH ₂ CH ₂ ) _m CH ₃ or polyamine chain -

such as spermine or spermidine;

where δ=0-2, ψ=1-3,

R ²⁶ in OR ²⁶ is optionally substituted alkyl, wherein the optional substituents are OH, halogen or NH ₂ ; A

m represents an integer from 1-8,

R ³ and R ⁴ of formula (XXVII) and (XXVIII) are independently selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted arylalkoxy, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted heteroarylalkyl or optionally substituted heterocycloalkyl, where the substituents are alkyl, halogen or OH;

R ⁵ , R ⁶ , R ⁷ and R ⁸ of formula (XXVII) and (XXVIII) are independently selected from hydrogen, optionally substituted alkyl or optionally substituted cycloalkyl;

R31 of formulas (XXVII) and (XXVIII) is selected from alkyl, aryl, arylalkyl, heteroaryl or optionally further substituted heteroarylalkyl, preferably selected from the group consisting of:

X of formulas (XXVII) and (XXVIII) is selected from -(CR ⁸¹ R ⁸² ) _m -, optionally substituted heteroaryl or heterocyclyl,

Z of formulas (XXVII) is selected from C=O, -O-, -NR, -CONH-, -NHCO- or may be absent;

R ⁸¹ and R ⁸² in -(CR ⁸¹ R ⁸² ) _m are independently selected from hydrogen, halogen, alkyl or cycloalkyl, or R ⁸¹ and R ⁸² can be taken together to form a carbocyclic ring;

R ¹⁰ and R ¹¹ in independently selected from hydrogen, halogen or alkyl;

R ¹² , R ¹³ , R ¹⁴ , R ¹⁵ and R ¹⁶ in

And independently selected from hydrogen, halogen or optionally substituted alkyl or OR ¹⁷ ;

R ¹⁷ is selected from hydrogen, optionally substituted alkyl or optionally substituted cycloalkyl;

m and n in -(CR ²¹ R ²² ) _m - and are independently O, 1,2,3 or 4;

o and r in are independently 0,1,2 or 3;

q and t in

are independently 0, 1, 2, 3 or 4;

g in represents 0 or 1;

and/or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[0098] In any of the compounds described herein, the ILM may have the structure of formulas (XXIX), (XXX), (XXXI) or (XXXII), which are derivatives of the IAP ligands described in WO publication No. 2014/055461 and Kim, KS , Discovery of tetrahydroisoquinoline-based bivalent heterodimeric IAP antagonists. Bioorg. Med. Chem. Lett. 24(21), 5022-9 (2014) or its non-natural mimetic, and the chemical linker of the linker group L is as shown:

And

Where:

R ² of formulas (XXIX) to (XXXII) is selected from H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl, or optionally substituted aryl; or alternatively;

R ¹ and R ² of formula (XXVII) and (XXVIII) are independently selected from H, optionally substituted thioalkyl -CR ⁶⁰ R ⁶¹ SR ⁷⁰ wherein R ⁶⁰ and R ⁶¹ are selected from H or methyl, and R ⁷⁰ is optionally substituted alkyl, optionally substituted branched alkyl, optionally substituted heterocyclyl, -(CH ₂ ) _v COR ²⁰ , -CH ₂ CHR ²¹ COR ²² or -CH ₂ R ²³ ; Where:

v is an integer from 1-3;

R ²⁰ and R ²² in -(CH ₂ ) _v COR ²⁰ and -CH ₂ CHR ²¹ COR ²² are independently selected from OH, NR ²⁴ R ²⁵ or OR ²⁶ ;

R ²¹ in -CH ₂ CHR ²¹ COR ²² selected from NR ²⁴ R ²⁵ ;

R ²³ in -CH ₂ R ²³ is selected from optionally substituted aryl or optionally substituted heterocyclyl, where optional substituents include alkyl and halogen;

R ²⁴ in NR ²⁴ R ²⁵ is selected from hydrogen or optionally substituted alkyl;

R ²⁵ in NR ²⁴ R ²⁵ is selected from hydrogen, optionally substituted alkyl, optionally substituted branched alkyl, optionally substituted arylalkyl, optionally substituted heterocyclyl, -CH ₂ CH ₂ (OCH ₂ CH ₂ ) _m CH ₃ or polyamine chain - such as spermine or spermidine,

where δ=0-2, ψ=1-3,

R ²⁶ in OR ²⁶ is optionally substituted alkyl, wherein the optional substituents are OH, halogen or NH ₂ ;

m is an integer from 1-8;

R ⁶ and R ⁸ of formula (XXIX) to (XXXII) are independently selected from hydrogen, optionally substituted alkyl or optionally substituted cycloalkyl; And

R ³¹ of formulas (XXIX) to (XXXII) is selected from alkyl, aryl, arylalkyl, heteroaryl or optionally further substituted heteroarylalkyl, preferably selected from the group consisting of:

[0099] In certain embodiments, the connection ILM is:

[00100] In any of the compounds described herein, the ILM may have a structure of formula (XXXIII), which is a derivative of the IAP ligands described in WO Publication No. 2014/074658 and WO Publication No. 2013/071035, or a non-natural mimetic thereof:

Where:

R ² of formula (XXXIII) is selected from H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclyl, optionally substituted arylalkyl, or optionally substituted aryl;

R ⁶ and R ⁸ of formula (XXXIII) are independently selected from hydrogen, optionally substituted alkyl, or optionally substituted cycloalkyl;

R ³² of formula (XXXIII) is selected from (C ₁ -C ₄ alkylene)-R ³³ , where R ³³ is selected from hydrogen, aryl, heteroaryl or cycloalkyl, optionally further substituted;

X formula (XXXIII) selected from:

Z and Z' of formula (XXXIII) are independently selected from:

where is everyone displays the connection attachment point, a Z and Z'

both can't be in any given connection;

Y of formula (XXXIII) is selected from:

And

where Z and Z’ of formula (XXXIII) are the same and Z

represents where is everyone displays the connection attachment point, X selected from:

And

And

Y of formula (XXXIII) is independently selected from:

And

Where:

displays the point of attachment to the -C=O part of the connection;

displays the point of attachment to the -NH part of the compound;

displays the first Z attachment point;

displays the second Z attachment point;

m is an integer from 0-3;

n is an integer from 1-3;

p is an integer from 0-4; And

A represents -C(O)R ³ ;

R ³ is selected from -C(O)R ³ is OH, NHCN, NHSO ₂ R ¹⁰ , NHOR ¹¹ or N(R ¹² )(R ¹³ );

R ¹⁰ and F ¹¹ in NHSO ₂ R ¹⁰ and NHOR ¹¹ are independently selected from hydrogen, optionally substituted by -C ₁ -C ₄ alkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl or heterocycloalkyl;

R ¹² and R ¹³ in N(R ¹² )(R ¹³ ) are independently selected from hydrogen, -C ₁ -C ₄ alkyl, -(C ₁ -C ₄ ) alkylene)--(C ₁ -C ₄ alkyl) and - (C ₁ -C ₄ alkylene)-O-(C ₁ -C ₄ hydroxyalkyl), or R ¹² and R ¹³ are taken together with the nitrogen atom to which they are usually added to form a saturated heterocyclyl, optionally containing one additional heteroatom selected of N, O and S, and wherein the saturated heterocycle is optionally substituted with methyl.

[00101] In any of the compounds described herein, the ILM may have the structure of formulas (XXXIV) or (XXXV), which are derivatives of the IAP ligands described in WO publication No. 2014/047024, or a non-natural mimetic thereof:

or

Where:

X of formula (XXXIV) or (XXXV) is absent or the group is selected from -(CR ¹⁰ R ¹¹ ) _m -, optionally substituted heteroaryl or optionally substituted heterocyclyl,

Y and Z of formula (XXXIV) or (XXXV) are independently selected from C=0, -0-, -NR ⁹ -, -CONH-, -NHCO- or may be absent;

R ¹ and R ² of formula (XXXIV) or (XXXV) are independently selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted arylalkyl, optionally substituted aryl, or

R ¹ and R ² of formula (XXXIV) or (XXXV) are independently selected from optionally substituted thioalkyl, wherein the substituents attached to the S atom of the thioalkyl are optionally substituted alkyl, optionally substituted branched alkyl, optionally substituted heterocyclyl, -(CH ₂ ) _v COR ²⁰ , -CH ₂ CHR ²¹ COR ²² or -CH ₂ R ²³ ;

where v is an integer from 1-3;

R ²⁰ and R ²² in -(CH ₂ ) _v COR ²⁰ and -CH ₂ CHR ²¹ COR ²² are independently selected from OH, NR ²⁴ R ²⁵ or OR ²⁶ ;

R ²¹ in -CH ₂ CHR ²¹ COR ²² selected from NR ²⁴ R ²⁵ ;

R ²³ in -CH ₂ R ²³ is selected from optionally substituted aryl or optionally substituted heterocyclyl, where optional substituents include alkyl and halogen;

R ²⁴ in NR ²⁴ R ²⁵ is selected from hydrogen or optionally substituted alkyl;

R ²⁵ in NR ²⁴ R ²⁵ is selected from hydrogen, optionally substituted alkyl, optionally substituted branched alkyl, optionally substituted arylalkyl, optionally substituted heterocyclyl, -CH ₂ (OCH ₂ CH ₂₀ ) _m CH ₃ or a polyamine chain;

R ²⁶ represents optionally substituted alkyl, where optional substituents are OH, halogen or NH ₂ ;

m in -(CR ¹⁰ R ¹¹ ) _m - represents an integer from 1-8;

R ³ and R ⁴ of formula (XXXIV) or (XXXV) are independently selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted arylalkoxy, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted heteroarylalkyl or optionally substituted heterocycloalkyl, where the substituents are alkyl, halogen or OH;

R ⁵ , R ⁶ , R ⁷ and R ⁸ of formula (XXXIV) or (XXXV) are independently selected from hydrogen, optionally substituted alkyl or optionally substituted cycloalkyl;

R ¹⁰ and R ¹¹ in -(CR ¹⁰ R ¹¹ ) _m are independently selected from hydrogen, halogen or optionally substituted alkyl;

R ¹² and R ¹³ in independently selected from hydrogen, halogen or optionally substituted alkyl, or R ¹² and R ¹³ can be taken together to form a carbocyclic ring;

R ¹⁴ , R ¹⁵ , R ¹⁶ , R ¹⁷ and R ¹⁸ in

And independently selected from hydrogen, halogen, optionally substituted alkyl or OR ¹⁹ ;

R ¹⁹ in OR ¹⁹ is selected from hydrogen, optionally substituted alkyl, or optionally substituted cycloalkyl;

m and n in -(CR ¹⁰ R ¹¹ ) _m - are independently 0, 1, 2, 3 or 4;

o and p in -(CR ¹⁰ R ¹¹ ) _m - are independently 0, 1, 2 or 3;

q in -(CR ¹⁰ R ¹¹ ) _m - represents 0, 1, 2, 3 or 4; r is 0 or 1;

t in -(CR ¹⁰ R ¹¹ ) _m - represents 1, 2 or 3; and/or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[00102] In any of the compounds described herein, the ILM may have a structure of formula (XXXVI), which is a derivative of the IAP ligands described in WO publication No. 2014/025759, or a non-natural mimetic thereof:

Where:

And formula (XXXVI) is selected from: or where the dotted line represents an optional double bond;

X of formula (XXXVI) is selected from: -(CR ²¹ R ²² ) _m -,

Y and Z of formula (XXXVI) are independently selected from -O-, -NR ⁶ - or absent;

V of formula (XXXVI) is selected from -N- or -CH-;

W of formula (XXXVI) is selected from -CH- or -N-;

R ¹ of formula (XXXVI) is selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted arylalkyl, or optionally substituted aryl;

R ³ and R ⁴ of formula (XXXVI) are independently selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted arylalkyl, optionally substituted heteroarylalkyl, or optionally substituted heterocycloalkyl;

R ⁵ , R ⁶ , R ⁷ and R ⁸ of formula (XXIV), (XXV) or (XXVI) are independently selected from hydrogen, optionally substituted alkyl or optionally substituted cycloalkyl, or preferably methyl;

R ⁹ and R ¹⁰ in independently selected from hydrogen, halogen or optionally substituted alkyl, or R ⁹ and R ¹⁰ can be taken together to form a carbocyclic ring;

R^eleven, R¹², R¹³ and R¹⁴ V independently selected from hydrogen, halogen, optionally substituted alkyl or OR¹⁵;

R ¹⁵ in OR ¹⁵ is selected from hydrogen, optionally substituted alkyl, or optionally substituted cycloalkyl;

m and n in- (CR ²¹ R ²² ) _m - and independently selected from 0, 1, 2, 3 or 4;

o and r in and independently selected from 0, 1,2 or 3;

q in or selected from 0, 1, 2, 3 or 4;

r in selected from 0 or 1, and/or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[00103] In any of the compounds described herein, the ILM may have the structure of formulas (XXXVII) or (XXXVIII), which are derivatives of the IAP ligands described in WO publication No. 2014/011712, or a non-natural mimetic thereof:

Where:

X of formulas (XXXVII) and (XXXVIII) is -(CR ¹⁶ R ¹⁷ ) _m -,

or absent;

Y and Z of formula (XXXVII) n (XXXVIII) are independently selected from -0-, C=0, NR ⁶ or none;

R ¹ and R ² of formula (XXXVII) and (XXXVIII) are selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkylaryl, or optionally substituted aryl;

R ³ and R ⁴ of formula (XXXVII) and (XXXVIII) are independently selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted arylalkyl, or optionally substituted aryl;

R ⁵ and R ⁶ of formula (XXXVII) and (XXXVIII) are independently selected from optionally substituted alkyl or optionally substituted cycloalkyl;

R ⁷ and R ⁸ of formula (XXXVII) and (XXXVIII) are independently selected from hydrogen, optionally substituted alkyl or optionally substituted cycloalkyl, or preferably methyl;

R ⁹ and R ¹⁰ in independently selected from hydrogen, optionally substituted alkyl, or R ⁹ and R ¹⁰ may be taken together to form a ring;

from R ¹¹ to R ¹⁴ in independently selected from hydrogen, halogen, optionally substituted alkyl or OR ¹⁵ ;

R ¹⁵ in OR ¹⁵ is selected from hydrogen, optionally substituted alkyl, or optionally substituted cycloalkyl;

R ¹⁶ and R ¹⁷ in -(CR ¹⁶ R ¹⁷ ) _m are independently selected from hydrogen, halogen or optionally substituted alkyl;

R ⁵⁰ and R ⁵¹ of formula (XXXVII) n (XXXVIII) are independently selected from optionally substituted alkyl, or R ⁵⁰ and R ⁵¹ are taken together to form a carbocyclic ring;

m and n in -(CR ¹⁶ R ¹⁷ ) _m - and independently represent an integer from 0-4;

o and r in independently represent an integer from 0-3;

q in represents an integer from 0-4; And

r in represents an integer from 0-1;

or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.

[00104] In an embodiment, R ¹ and R ² ILM of formula (XXXVII) or (XXXVIII) are tert-butyl, and R ³ and R ⁴ ILM of formula (XXXVII) or (XXXVIII) are tetrahydronaphthalene.

[00105] In any of the compounds described herein, the ILM may have the structure of formulas (XXXLX) or (XL), which are derivatives of the IAP ligands described in WO publication No. 2013/071039, or a non-natural mimetic thereof:

Where:

R ⁴³ and R ⁴⁴ of formulas (XXXIX) and (XL) are independently selected from hydrogen, alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, cycloalkylalkyl, further optionally substituted, and

R ⁶ and R ⁸ of formula (XXXIX) and (XL) are independently selected from hydrogen, optionally substituted alkyl, or optionally substituted cycloalkyl.

Each X of formulas (XXXIX) and (XL) is independently selected from:

And

each Z of formulas (XXXIX) and (XL) is selected from where is everyone displays the connection attachment point; and each Y is selected from:

Where:

displays the point of attachment to the -C=O part of the connection;

displays the point of attachment to the amino part of the compound;

displays the first Z attachment point;

displays the second Z attachment point; and A is selected from -C(O)R ³ or

or a tautomeric form of any of the above, in which:

R ³ in -C(O)R ³ is selected from OH, NHCN, NHSO ₂ R ¹⁰ , NHOR ¹¹ or N(R ¹² )(R ¹³ );

R ¹⁰ and R ¹¹ in NHSO ₂ R ¹⁰ and NHOR ¹¹ are independently selected from -C ₁ -C ₄ alkyl, cycloalkyl, aryl, heteroaryl or heterocycloalkyl, any of which is optionally substituted, and hydrogen;

each of R ¹² and R ¹³ in N(R ¹² )(R ¹³ ) is independently selected from hydrogen, -C ₁ -C ₄ alkyl, -(C ₁ -C ₄ alkylene)-NH-(C ₁ -C ₄ alkyl) , benzyl, -(C ₁ -C ₄ alkylene)-C(O)OH,

-(C ₁ -C ₄ alkylene)-C(O)CH ₃ , -CH(benzyl)-COOH, -C ₁ -C ₄ alkoxy and

-(C ₁ -C ₄ alkylene)-O-(C ₁ -C ₄ hydroxyalkyl); or R ¹² and R ¹³ in N(R ¹² )(R ¹³ ) are taken together with the nitrogen atom to which they are usually attached to form a saturated heterocyclyl optionally containing one additional heteroatom selected from N, O and S, and where saturated the heterocycle is optionally substituted with methyl.

[00106] In any of the compounds described herein, the ILM may have a structure of formula (XLI), which is a derivative of the IAP ligands described in WO publication No. 2013/071039, or a non-natural mimetic thereof:

Where:

W ¹ of formula (XLI) is selected from O, S, NR ^A or C(R ^8a )(R ^8b );

W ² of formula (XLI) is selected from O, S, NR ^A or C(R ^8c )(R ^8d ); provided that W ¹ and W ² are both not O, or both are not S;

R ¹ of formula (XLI) is selected from H, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, -C ₁ -C ₆ alkyl (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl), or -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl);

when X ¹ is selected from O, NR ^A , S, S(O), or S(O) ₂ then X ² is C(R ^2a R ^2b );

or:

X ¹ of formula (XLI) is selected from CR ^2c R ^2d and X ² is CR ^2a R ^2b , and R ^2c and R ^2a together form a bond;

or:

X ¹ and X ² of formula (XLI) are independently selected from C and N and are members of a fused substituted or unsubstituted saturated or partially saturated 3-10 membered cycloalkyl ring, fused substituted or unsubstituted saturated or partially saturated 3-10 membered heterocycloalkyl ring, fused substituted or an unsubstituted 5-10 membered aryl ring or a fused substituted or unsubstituted 5-10 membered heteroaryl ring; or:

X ¹ of formula (XLI) is selected from CH ₂ and X ² is C=0, C=C( ^RC ) ₂ , or C=NR ^C ; where each R ^c is independently selected from H, -CN, -OH, alkoxy, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl) or -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl);

R ^A in NR ^A is selected from H, C ₁ -C ₆ alkyl, -C(=O)C ₁ -C ₂ alkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;

R ^2a , R ^2b , R ^2c , R ^2d in CR ^2c R ^2d and CR ^2a R ^2b are independently selected from H, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted _{C 1} -C ₆ heteroalkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl-(substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl) and - C(=O )R ^B ;

R ^B in - C(=O)R ^B is selected from substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl), -C ₁ -C ₆ alkyl - (substituted or unsubstituted aryl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl) or - NR ^D R ^E ;

R ^D and R ^E in NR ^D R ^E are independently selected from H, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl) or -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl);

m of formula (XLI) is selected from 0, 1 or 2;

-U- formula (XLI) is selected from -NHC(=O)-, -C(=O)NH-, -NHS(=O) ₂ -, -S(=O) ₂ NH-, -NHC(=O )NH-, -NH(C=O)O-, -O(C=O)NH- or -NHS(=O) ₂ NH-;

R ³ of formula (XLI) is selected from C ₁ -C ₃ alkyl or C ₁ -C ₃ fluoroalkyl;

R ⁴ of formula (XLI) is selected from -NHR ⁵ , -N(R ⁵ )2, -N+(R ⁵ )3 or -OR ⁵ ;

each R ⁵ in -NHR ⁵ , -N(R ⁵ )2, -N+(R ⁵ )3 and -OR ⁵ is independently selected from H, C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, C ₁ -C ₃ heteroalkyl and -C ₁ -C ₃ alkyl-(C ₃ -C ₅ cycloalkyl);

or:

R ³ and R ⁵ of formula (XLI), together with the atoms to which they are attached, form a substituted or unsubstituted 5-7 membered ring;

or:

R ³ of formula (XLI) is bonded to the U nitrogen atom to form a substituted or unsubstituted 5-7 membered ring;

R ⁶ of formula (XLI) is selected from -NHC(=O)R ⁷ , -C(=O)NHR ⁷ , -NHS(=O) ₂ R ⁷ , -S(=O) ₂ NHR ⁷ ; -NHC(=O)NHR ⁷ , -NHS(=O) ₂ NHR ⁷ , -(C ₁ -C ₃ alkyl)-NHC(=O)R ⁷ , -(C ₁ -C ₃ alkyl)-C(= O)NHR ⁷ , -(C ₁ -C ₃ alkyl)-NHS(=O) ₂ R ⁷ , -(C ₁ -C ₃ alkyl)-S(=O) ₂ NHR ⁷ ; -(C ₁ -)-NHC(=O)NHR ⁷ , -(C ₁ -)-NHS(=O) ₂ NHR ⁷ , substituted or unsubstituted C ₂ -C ₁₀ heterocycloalkyl or substituted or unsubstituted heteroaryl;

each R ⁷ in -NHC(=O)R ⁷ , -C(=O)NHR ⁷ , -NHS(=O) ₂ R ⁷ , -S(=O) ₂ NHR ⁷ ; -NHC(=O)NHR ⁷ , -NHS(=O) ₂ NHR ⁷ , -(C ₁ -)-NHC(=O)R ⁷ , -(C ₁ -C ₃ alkyl)-C(=O)NHR ⁷ , -(C ₁ -)-NHS(=O) ₂ R ⁷ , -(C ₁ -)-S(=O) ₂ NHR ⁷ ; -(C ₁ -)-NHC(=O)NHR ⁷ , -(C ₁ -)-NHS(=O) ₂ NHR ⁷ independently selected from C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ - C ₆ heteroalkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₁ heterocycloalkyl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl), -( _CH2 )p-CH(substituted or unsubstituted aryl)2, -( _CH2 ) _p -CH(substituted or unsubstituted heteroaryl)2, -( _CH2 ) _p -CH(substituted or unsubstituted aryl)( substituted or unsubstituted heteroaryl), -(substituted or unsubstituted aryl)-(substituted or unsubstituted aryl), -(substituted or unsubstituted aryl)-(substituted or unsubstituted heteroaryl), -(substituted or unsubstituted heteroaryl)-(substituted or unsubstituted aryl) or -(substituted or unsubstituted heteroaryl)-(substituted or unsubstituted heteroaryl); p in R ⁷ is selected from 0, 1 or 2;

R ^8a , R ^8b , R ^8c , and R ^8d in C(R ^8a )(R ^8b ) and C(R ^8c )(R ^8d ) are independently selected from H, C ₁ -C ₆ alkyl, C ₁ -C ₆ fluoroalkyl , C ₁ -C ₆ alkoxy, C ₁ -C ₆ heteroalkyl and substituted or unsubstituted aryl

or:

R ^8a and R ^8d are as defined above, and R ^8b and R ^8c together form a bond;

or:

R ^8a and R ^8d are as defined above, and R ^8b and R ^8c together with the atoms to which they are attached form a substituted or unsubstituted fused 5-7 membered saturated or partially saturated carbocyclic ring or heterocyclic ring containing 1-3 heteroatoms , selected from S, O and N, a substituted or unsubstituted fused 5-10 membered aryl ring, or a substituted or unsubstituted fused 5-10 membered heteroaryl ring containing 1-3 heteroatoms selected from S, O and N; or:

R ^8c and R ^8d are as defined above, and R ^8a and R ^8b together with the atoms to which they are attached form a substituted or unsubstituted, saturated or partially saturated 3-7 membered spirocycle or heterospirocycle containing 1-3 heteroatoms selected from S, O and N; or:

R ^8a and R ^8b are as defined above, and R ^8c and R ^8d together with the atoms to which they are attached form a substituted or unsubstituted, saturated or partially saturated 3-7 membered spirocycle or heterospirocycle containing 1-3 heteroatoms selected from S, O and N;

wherein each substituted alkyl, heteroalkyl, fused ring, spirocycle, heterospirocycle, cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with 1-3 R ⁹ ; n

each R ⁹ in R ^8a , R ^8b , R ^8c n R ^8d is independently selected from halogen, -OH, - SH, (C=O), CN, C ₁ -C ₄ alkyl, C1-C4fluoroalkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ fluoroalkoxy, -NH ₂ , -NH(C ₁ -C ₄ alkyl), -NH(C ₁ -C ₄ alkyl) ₂ , - C(=O)OH, -C(=O) ₂ H ₂ , -C(=O)C ₁ -C ₃ alkyl, -S(=O) ₂ CH ₃ , -NH(C ₁ -C ₄ alkyl)-OH, -NH(C ₁ -C ₄ alkyl) -O-(C ₁ -C ₄ alkyl), -O(C ₁ -C ₄ alkyl)-NH2; -O(C ₁ -C ₄ alkyl)-NH-(C ₁ -C ₄ alkyl) and -O(C ₁ -C ₄ alkyl)-N-(C ₁ -C ₄ alkyl) ₂ , or two R ⁹ together with the atoms to which they are attached form a methylenedioxy or ethylenedioxy ring, substituted or unsubstituted with halogen, OH or C ₁ -C ₃ alkyl.

[00107] In any of the compounds described herein, the ILM may have a structure of formula (XLII), which is a derivative of the TAP ligands described in WO publication No. 2013/071039, or a non-natural mimetic thereof:

Where:

W ¹ of formula (XLII) represents O, S, NR ^A or C(R ^8a )(R ^8b );

W ² of formula (XLII) is O, S, NR ^A or C( ^R8c )( ^R8d ); provided that W ¹ and W ² are both not O, or both are not S;

R ¹ of formula (XLII) is selected from H, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), substituted or unsubstituted arnl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl) or -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl);

when X ¹ of formula (XLII.) is NR ^A , then X ² is C=O, or CR ^2c R ^2d , and X ³ is CR ^2a R ^2b ;

or:

when X ¹ of formula (XLII) is selected from S, S(O), or S(O) ₂ , then X ² is CR ^2c R ^2d and X ³ is CR ^2a R ^2b ;

or:

when X ¹ of formula (XLII) represents O, then X ² represents CR ^2c R ^2d and NR ^A and X ³ represents CR ^2a R ^2b ;

or:

when X ¹ of formula (XLII) is CH ₃ , then X ² is selected from O, NR ^A , S, S(O) or S(O) ₂ and X ³ is CR ^2a R ^2b ;

when X ¹ of formula (XLII) is CR ^2e R ^2f and X ² is CR ^2c R ^2d , and R ^2e and R ^2c together form a bond, and X ³ of formula (VLII) is CR ^2a R ^2b ; or:

X ¹ and X ³ of formula (XLII) are both CH ₂ and X ² of formula (XLII) is C=O, C=C(RC) ₂ , or C=NRC; where each R ^C is independently selected from H, -CN, -OH, alkoxy, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₃ heterocycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl) or -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl); or:

X ¹ and X ² of formula (XLII) are independently selected from C and N, and are members of a fused substituted or unsubstituted saturated or partially saturated 3-10 membered cycloalkyl ring, fused substituted or unsubstituted saturated or partially saturated 3-10 membered heterocycloalkyl ring, fused a substituted or unsubstituted 5-10 membered aryl ring or a fused substituted or unsubstituted 5-10 membered heteroaryl ring, and X ³ represents CR ^2a R ^2b ; or:

X ² and X ³ of formula (XLII) are independently selected from C and N, and are members of a fused substituted or unsubstituted saturated or partially saturated 3-10 membered cycloalkyl ring, fused substituted or unsubstituted saturated or partially saturated 3-10 membered heterocycloalkyl ring, fused a substituted or unsubstituted 5-10 membered aryl ring, or a fused substituted or unsubstituted 5-10 membered heteroaryl ring, and X ¹ of formula (VLII) is CR ^2e R ^2f ;

R ^A in NR ^A is selected from H, C ₁ -C ₆ alkyl, -C(=O)C ₁ -C ₂ alkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R ^2a , R ^2b , R ^2c , R ^2d , R ^2e , and R ^2f in CR ^2c R ^2d , CR ^2a R ^2b and CR ^2e R ^2f are independently selected from H, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ heteroalkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl), -C ₁ -C ₆ alkyl -(substituted or unsubstituted heteroaryl) and - C(=O)R ^B ; R ^B in -C(=O)R ^B is selected from substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl), -C ₁ -C ₆ alkyl - (substituted or unsubstituted aryl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl) or - NR ^D R ^E ;

R ^D and R ^E in NR ^D R ^E are independently selected from H, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ - C ₅ heterocycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl) or -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl);

m of formula (XLII) is selected from 0, 1 or 2;

-U- of formula (XLII) is selected from -NHC(=O)-, -C(=O) ₂ H-, -NHS(=O) ₂ -, -S(=O) ₂ NH-, -NHC(= O) ₂ H-, -NH(C=O)O-, -O(C=O) ₂ H- or -NHS(=O) ₂ NH-;

R ³ of formula (XLII) is selected from C ₁ -C ₃ alkyl or C ₁ -C ₃ fluoroalkyl;

R ⁴ of formula (XLII) is selected from -NHR ⁵ , -N(R ⁵ ) ₂ , -N+(R ⁵ )3 or -OR ⁵ ;

each R ⁵ in -NHR ⁵ , -N(R ⁵ ) ₂ , -N+(R ⁵ )3 and -OR ⁵ is independently selected from H, C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, C ₁ -C ₃ heteroalkyl and -C ₁ -C ₃ alkyl-(C ₃ -C ₅ cycloalkyl);

or:

R ³ and R ⁵ of formula (XLII), together with the atoms to which they are attached, form a substituted or unsubstituted 5-7 membered ring;

or:

R ³ of formula (XLII) is bonded to the nitrogen atom U to form a substituted or unsubstituted 5-7 membered ring;

R ⁶ of formula (XLII) is selected from -NHC(=O)R ⁷ , -C(=O) ₂ HR ⁷ , -NHS(=O) ₂ R ⁷ , -S(=O) ₂ NHR ⁷ ; -NHC(=O) ₂ HR ⁷ , -NHS(=O) ₂ NHR ⁷ , -(C ₁ -C ₃ alkyl)-NHC(=O)R ⁷ , -(C ₁ -C ₃ alkyl)-C( =O) ₂ NHR ⁷ , -(C ₁ -C ₃ alkyl)-NHS(=O) ₂ R ⁷ , -(C ₁ -C ₃ alkyl)-S(=O) ₂ NHR ⁷ ; -(C ₁ -C ₃ alkyl)-NHC(=O) ₂ HR ⁷ , -(C ₁ -C ₃ alkyl)-NHS(=O) ₂ NHR ⁷ , substituted or unsubstituted C ₂ -C ₁₀ heterocycloalkyl or substituted or unsubstituted heteroaryl;

each R ⁷ in -NHC(=O)R ⁷ , -C(=O) ₂ HR ⁷ , -NHS(=O) ₂ R ⁷ , -S(=O) ₂ NHR ⁷ ; -NHC(=O) ₂ HR ⁷ , -NHS(=O) ₂ NHR ⁷ , -(C ₁ -C ₃ alkyl)-NHC(=O)R ⁷ , -(C ₁ -C ₃ alkyl)-C( =O) ₂ HR ⁷ , -(C ₁ -C ₃ alkyl)-NHS(=O) ₂ R ⁷ , -(C ₁ -C ₃ alkyl)-S(=O) ₂ NHR ⁷ ; -(C ₁ -C ₃ alkyl)-NHC(=O) ₂ HR ⁷ , -(C ₁ -C ₃ alkyl)-NHS(=O) ₂ NHR ⁷ independently selected from C ₁ -C ₆ alkyl, C ₁ - C ₆ haloalkyl, C ₁ -C ₆ heteroalkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl-( substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₁₀ heterocycloalkyl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl), -C ₁ -C ₆ alkyl-(substituted or unsubstituted heteroaryl), -(CH ₂ ) _p -CH (substituted or unsubstituted aryl) ₂ , -(CH ₂ ) _p -CH (substituted or unsubstituted heteroaryl) ₂ , -(CH ₂ ) _p -CH (substituted or unsubstituted aryl)(substituted or unsubstituted heteroaryl), -(substituted or unsubstituted aryl)-(substituted or unsubstituted aryl), -(substituted or unsubstituted aryl)-(substituted or unsubstituted heteroaryl), -(substituted or unsubstituted heteroaryl) -(substituted or unsubstituted aryl) or -(substituted or unsubstituted heteroaryl)-(substituted or unsubstituted heteroaryl);

p in R ⁷ is selected from 0, 1 or 2;

R ^8a , R ^8b , R ^8c , and R ^8d in C(R ^8a )(R ^8b ) and C(R ^8c )(R ^8d ) are independently selected from H, C ₁ -C ₆ alkyl, C ₁ -C ₆ fluoroalkyl , C ₁ -C ₆ alkoxy, C ₁ -C ₆ heteroalkyl and substituted or unsubstituted aryl; or:

R ^8a and R ^8d are as defined above, and R ^8b and R ^8c together form a bond;

or:

R ^8a and R ^8d are as defined above, and R ^8b and R ^8c together with the atoms to which they are attached form a substituted or unsubstituted fused 5-7 membered saturated or partially saturated carbocyclic ring or heterocyclic ring containing 1-3 heteroatoms , selected from S, O and N, a substituted or unsubstituted fused 5-10 membered aryl ring, or a substituted or unsubstituted fused 5-10 membered heteroaryl ring containing 1-3 heteroatoms selected from S, O and N; or:

R ^8c and R ^8d are as defined above, and R ^8a and R ^8b together with the atoms to which they are attached form a substituted or unsubstituted, saturated or partially saturated 3-7 membered spirocycle or heterospirocycle containing 1-3 heteroatoms selected from S, O and N; or:

R ^8a and R ^8b are as defined above, and R ^8c and R ^8d together with the atoms to which they are attached form a substituted or unsubstituted, saturated or partially saturated 3-7 membered spirocycle or heterospirocycle containing 1-3 heteroatoms selected from S, O and N; wherein each substituted alkyl, heteroalkyl, fused ring, spirocycle, heterospirocycle, cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with 1 -3 R ⁹ ; And

each R ⁹ in R ^8a , R ^8b , R ^8c and R ^8d is independently selected from halogen, -OH, -SH, (C=O), CN, C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ fluoroalkoxy, -NH ₂ , -NH(C ₁ -C ₄ alkyl), -NH(C ₁ -C ₄ alkyl) ₂ , - C(=O)OH, -C( =O) ₂ H ₂ , -C(=O)C ₁ -C ₃ alkyl, -S(=O) ₂ CH ₃ , -NH(C ₁ -C ₄ alkyl)-OH, -NH(C ₁ -C ₄ alkyl)-O-(C-C ₄ alkyl), -O(C ₁ -C ₄ alkyl)-NH ₂ ; -O(C ₁ -C ₄ alkyl)-NH-(C ₁ -C ₄ alkyl) and -O(C ₁ -C ₄ alkyl)-N-(C ₁ -C ₄ alkyl) ₂ , or two R ⁹ together with the atoms to which they are attached form a methylenedioxy or ethylenedioxy ring, substituted or unsubstituted with halogen, -OH or C ₁ -C ₃ alkyl.

[00108] In any of the compounds described herein, the ILM may have a structure of formula (XLIII), which is a derivative of the IAP ligands described in WO publication No. 2013/071039, or a non-natural mimetic thereof:

Where:

W ¹ of formula (XLIII) is selected from O, S, NR ^A , or C(R ^8a )(R ^8b );

W ² of formula (XLIII) is selected from O, S, NR ^A or C(R ^8c )( ^R8d ); provided that W ¹ and W ² are both not O, or both are not S;

R ¹ of formula (XLIII) is selected from H, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, -C ₁ -C ₆ alkyl (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl) or -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl);

when X ¹ of formula (XLIII) is selected from NR ^A , S, S(O) or S(O) ₂ , then X ² of formula (XLIII) is CR ^2c R ^2d and X ³ of formula (XLIII) is CR ^{2a R2b} ;

or:

when X ¹ of formula (XLIII) is O, then X ² of formula (XLIII) is selected from O, NR ^A , S, S(O) or S(O) ₂ and X ³ of formula (XLIII) is CR ^2a R ^2b ; or:

when X ¹ of formula (XLIII) is CR ^2e R ^2f and X ² of formula (XLIII) is CR ^2c R ^2d and R ^2e and R ^2c together form a bond and X ³ of formula (XLIII) is CR ^2a R ^2b ; or:

X ¹ and X ² of formula (XLIII) are independently selected from C and N, and are members of a fused substituted or unsubstituted saturated or partially saturated 3-10 membered cycloalkyl ring, fused substituted or unsubstituted saturated or partially saturated 3-10 membered heterocycloalkyl ring, fused a substituted or unsubstituted 5-10 membered aryl ring or a fused substituted or unsubstituted 5-10 membered heteroaryl ring, and X ³ of formula (XLIII) is CR ^2a R ^2b ; or:

X ² and X ³ of formula (XLIII) are independently selected from C and N, and are members of a fused substituted or unsubstituted saturated or partially saturated 3-10 membered cycloalkyl ring, fused substituted or unsubstituted saturated or partially saturated 3-10 membered heterocycloalkyl ring, fused a substituted or unsubstituted 5-10 membered aryl ring or a fused substituted or unsubstituted 5-10 membered heteroaryl ring, and X ¹ of formula (VLII) is CR ^2e R ^2f ;

R ^A in NR ^A is H, C ₁ -C ₆ alkyl, -C(=O)C ₁ -C ₂ alkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R ^2a , R ^2b , R ^2c , R ^2d , R ^2e and R ^2f in CR ^2c R ^2d , CR ^2a R ^2b and CR ^2e R ^2f are independently selected from H, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ heteroalkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl) and - C(=O)R ^B ; R ^B in -C(=O)R ^B is substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl), -C ₁ -C ₆ alkyl - (substituted or unsubstituted aryl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl) or - NR ^D R ^E ;

R ^D and R ^E in NR ^D R ^E are independently selected from H, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ - C ₅ heterocycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl) or -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl); m of formula (XLIII) is 0, 1 or 2;

-U- of formula (XLIII) is -NHC(=O) ₂ , -C(=O) ₂ H-, -NHS(=O) ₂ -, -S(=O) ₂ NH-, -NHC(= O) ₂ H-, -NH(C=O)O-, -O(C=O) ₂ H- or -NHS(=O) ₂ NH-; R ³ of formula (XLIII) represents C ₁ -C ₃ alkyl or C ₁ -C ₃ fluoroalkyl;

R ⁴ of formula (XLIII) is -NHR ⁵ , -N(R ⁵ ) ₂ , -N+(R ⁵ ) ₃ or -OR ⁵ ; each R ⁵ in -NHR ⁵ , -N(R ⁵ ) ₂ , -N+(R ⁵ ) ₃ and -OR ⁵ is independently selected from H, C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, C ₁ -C ₃ heteroalkyl and -C ₁ -C ₃ alkyl-(C ₃ -C ₅ cycloalkyl); or:

R ³ and R ⁵ of formula (XLIII), together with the atoms to which they are attached, form a substituted or unsubstituted 5-7 membered ring;

or:

R ³ of formula (XLIII) is bonded to the nitrogen atom U to form a substituted or unsubstituted 5-7 membered ring;

R ⁶ of formula (XLIII) is selected from -NHC(=O)R ⁷ , -C(=O) ₂ HR ⁷ , -NHS(=O) ₂ R ⁷ , -S(=O) ₂ NHR ⁷ ; -NHC(=O) ₂ HR ⁷ , -NHS(=O) ₂ NHR ⁷ , -(C ₁ -C ₃ alkyl)-NHC(=O)R ⁷ , -(C ₁ -C ₃ alkyl)-C( =O) ₂ HR ⁷ , -(C ₁ -C ₃ alkyl)-NHS(=O) ₂ R ⁷ , -(C ₁ -C ₃ alkyl)-S(=O) ₂ NHR ⁷ ; -(C ₁ -C ₃ alkyl)-NHC(=O) ₂ HR ⁷ , -(C ₁ -C ₃ alkyl)-NHS(=O) ₂ NHR ⁷ , substituted or unsubstituted C ₂ -C ₁₀ heterocycloalkyl or substituted or unsubstituted heteroaryl;

each R ⁷ in -NHC(=O)R ⁷ , -C(=O) ₂ HR ⁷ , -NHS(=O) ₂ R ⁷ , -S(=O) ₂ NHR ⁷ ; -NHC(=O) ₂ HR ⁷ , -NHS(=O) ₂ NHR ⁷ , -(C ₁ -C ₃ alkyl)-NHC(=O)R ⁷ , -(C ₁ -C ₃ alkyl)-C( =O) ₂ HR ⁷ , -(C ₁ -C ₃ alkyl)-NHS(=O) ₂ R ⁷ , -(C ₁ -C ₃ alkyl)-S(=O) ₂ NHR ⁷ ; -(C ₁ -C ₃ alkyl)-NHC(=O) ₂ HR ⁷ , -(C ₁ -C ₃ alkyl)-NHS(=O) ₂ NHR ⁷ independently selected from C ₁ -C ₆ alkyl, C ₁ - C ₆ haloalkyl, C ₁ -C ₆ heteroalkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl-( substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₁₀ heterocycloalkyl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl), -C ₁ -C ₆ alkyl-(substituted or unsubstituted heteroaryl), -(CH ₂ ) _p -CH (substituted or unsubstituted aryl) ₂ , -(CH ₂ ) _p -CH (substituted or unsubstituted heteroaryl) ₂ , -(CH ₂ ) _p -CH (substituted or unsubstituted aryl)(substituted or unsubstituted heteroaryl), -(substituted or unsubstituted aryl)-(substituted or unsubstituted aryl), -(substituted or unsubstituted aryl)-(substituted or unsubstituted heteroaryl), -(substituted or unsubstituted heteroaryl) -(substituted or unsubstituted aryl) or -(substituted or unsubstituted heteroaryl)-(substituted or unsubstituted heteroaryl); p in R ⁷ is 0, 1 or 2;

R ^8a , R ^8b , R ^8c , and R ^8d in C(R ^8a )(R ^8b ) and C(R ^8c )(R ^8d ) are independently selected from H, C ₁ -C ₆ alkyl, C ₁ -C ₆ fluoroalkyl , C ₁ -C ₆ alkoxy, C ₁ -C ₆ heteroalkyl and substituted or unsubstituted aryl; or:

R ^8a and R ^8d are as defined above, and R ^8b and R ^8c together form a bond;

or:

R ^8a and R ^8d are as defined above, and R ^8b and R ^8c together with the atoms to which they are attached form a substituted or unsubstituted fused 5-7 membered saturated or partially saturated carbocyclic ring or a heterocyclic ring containing 1-3 heteroatoms , selected from S, O and N, a substituted or unsubstituted fused 5-10 membered aryl ring, or a substituted or unsubstituted fused 5-10 membered heteroaryl ring containing 1-3 heteroatoms selected from S, O and N; or:

R ^8c and R ^8d are as defined above, and R ^8a and R ^8b together with the atoms to which they are attached form a substituted or unsubstituted, saturated or partially saturated 3-7 membered spirocycle or heterospirocycle containing 1-3 heteroatoms selected from S, O and N; or:

R ^8a and R ^8b are as defined above, and R ^8c and R ^8d together with the atoms to which they are attached form a substituted or unsubstituted, saturated or partially saturated 3-7 membered spirocycle or heterospirocycle containing 1-3 heteroatoms selected from S, O and N; wherein each substituted alkyl, heteroalkyl, fused ring, spirocycle, heterospirocycle, cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with 1 -3 R ⁹ ; And

each R ⁹ in R ^8a , R ^8b , R ^8c and R ^8d is independently selected from halogen, -OH, -SH, (C=O), CN, C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ fluoroalkoxy, -NH ₂ , -NH(C ₁ -C ₄ alkyl), -NH(C ₁ -C ₄ alkyl) ₂ , - C(=O)OH, -C( =O) ₂ H ₂ , -C(=O)C ₁ -C ₃ alkyl, -S(=O) ₂ CH3, -NH(C ₁ -C ₄ alkyl)-OH, -NH(C ₁ -C ₄ alkyl)-O-(C-C ₄ alkyl), -O(C ₁ -C ₄ alkyl)-NH ₂ ; -O(C ₁ -C ₄ alkyl)-NH-(C ₁ -C ₄ alkyl) and -O(C ₁ -C ₄ alkyl)-N-(C ₁ -C ₄ alkyl) ₂ , or two R ⁹ together with the atoms to which they are attached form a methylenedioxy or ethylenedioxy ring, substituted or unsubstituted with halogen, -OH or C ₁ -C ₃ alkyl.

[00109] In any of the compounds described herein, the ILM may have a structure of formula (XLIV), which is a derivative of the TAP ligands described in WO publication No. 2013/071039, or a non-natural mimetic thereof:

Where:

W ¹ of formula (XLIV) is selected from O, S, NR ^A or C(R ^8a )(R ^8b );

W ² of formula (XLIV) is selected from O, S, NR ^A or C(R ^8c )(R ^8d ); provided that

W ¹ and W ² are both not O, or both are not S;

W ³ of formula (XLIV) is selected from O, S, NR ^A or C(R ^8e )(R ^8f ), provided that the ring containing W ¹ , W ² and W ³ does not contain two adjacent oxygen atoms or sulfur atoms;

R ¹ of formula (XLIV) is selected from H, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl) or -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl);

when X ¹ of formula (XLIV) is O, then X ² of formula (XLIV) is selected from CR ^2c R ^2d and ^NRA and X ³ of formula (XLIV) is CR ^2a R ^2b ;

or:

when X ¹ of formula (XLIV) is CH ₂ then X ² of formula (XLIV) is selected from O, ^NRA , S, S(O) or S(O) ₂ and X ³ of formula (XLIV) is CR ^{2a R2b} ;

or:

when X ¹ of formula (XLIV) is CR ^2e R ^2f and X ² of formula (XLIV) is CR ^2c R ^2d , and R ^2e and R ^2c together form a bond and X ³ of formula (XLIV) is CR ^2a R ^2b ; or:

X ¹ and X ³ of formula (XLIV) are both CH ₂ and X ² of formula (XLII) is C=O, C=C(RC) ₂ or C=NRC; where each Rc is independently selected from H, -CN, -OH, alkoxy, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl , substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ - C ₅ heterocycloalkyl), -C ₁ - C ₆ alkyl- (substituted or unsubstituted aryl) or -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl); or:

X ¹ and X ² of formula (XLIV) are independently selected from C and N, and are members of a fused substituted or unsubstituted saturated or partially saturated 3-10 membered cycloalkyl ring, fused substituted or unsubstituted saturated or partially saturated 3-10 membered heterocycloalkyl ring, fused a substituted or unsubstituted 5-10 membered aryl ring or a fused substituted or unsubstituted 5-10 membered heteroaryl ring, and X ³ of formula (XLIV) is CR ^2a R ^2b ; or:

X ² and X ³ of formula (XLIV) are independently selected from C and N, and are members of a fused substituted or unsubstituted saturated or partially saturated 3-10 membered cycloalkyl ring, fused substituted or unsubstituted saturated or partially saturated 3-10 membered heterocycloalkyl ring, fused a substituted or unsubstituted 5-10 membered aryl ring or a fused substituted or unsubstituted 5-10 membered heteroaryl ring, and X ¹ of formula (XLIV) is CR ^2e R ^2f ;

R ^A in NR ^A is selected from H, C ₁ -C ₆ alkyl, -C(=O)C ₁ -C ₂ alkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R ^2a , R ^2b , R ^2c , R ^2d , R ^2e and R ^2f in CR ^2c R ^2d , CR ^2a R ^2b and CR ^2e R ^2f are independently selected from H, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ heteroalkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₃ heterocycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl) and - C(=O)R ^B ; R ^B in -C(=O)R ^B is selected from substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl), -C ₁ -C ₆ alkyl - (substituted or unsubstituted aryl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl) or - NR ^D R ^E ;

R ^D and R ^E in NR ^D R ^E are independently selected from H, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₆ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₅ heterocycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl) or -C ₁ -C ₆ alkyl- (substituted or unsubstituted heteroaryl);

m of formula (XLIV) is selected from 0, 1 or 2;

-U- formula (XLIV) is selected from -NHC(=O)-, -C(=O) ₂ H-, -NHS(=O) ₂ -, -S(=O) ₂ NH-, -NHC(= O) ₂ H-, -NH(C=O)O-, -O(C=O) ₂ H- or -NHS(=O) ₂ NH-;

R ³ of formula (XLIV) is selected from C ₁ -C ₃ alkyl or C ₁ -C ₃ fluoroalkyl;

R ⁴ of formula (XLIV) is selected from -NHR ⁵ , -N(R ⁵ ) ₂ , -N+(R ⁵ ) ₃ or -OR ⁵ ; each R ⁵ in -NHR ⁵ , -N(R ⁵ ) ₂ , -N+(R ⁵ ) ₃ and -OR ⁵ is independently selected from H, C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, C ₁ -C ₃ heteroalkyl and -C ₁ -C ₃ alkyl-(C ₃ -C ₅ cycloalkyl);

or:

R ³ and R ⁵ of formula (XLIV), together with the atoms to which they are attached,

form a substituted or unsubstituted 5-7 membered ring;

or:

R ³ of formula (XLIII) is bonded to the nitrogen atom U to form a substituted or unsubstituted 5-7 membered ring;

R ⁶ of formula (XLIII) is selected from -NHC(=O)R ⁷ , -C(=O) ₂ HR ⁷ , -NHS(=O) ₂ R ⁷ , -S(=O) ₂ NHR ⁷ ; -NHC(=O) ₂ HR ⁷ , -NHS(=O) ₂ NHR ⁷ , -(C ₁ -C ₃ alkyl)-NHC(=O)R ⁷ , -(C ₁ -C ₃ alkyl)-C( =O) ₂ HR ⁷ , -(C ₁ -C ₃ alkyl)-NHS(=O) ₂ R ⁷ , -(C ₁ -C ₃ alkyl)-S(=O) ₂ NHR ⁷ ; -(C ₁ -C ₃ alkyl)-NHC(=O) ₂ HR ⁷ , -(C ₁ -C ₃ alkyl)-NHS(=O) ₂ NHR ⁷ , substituted or unsubstituted C ₂ -C ₁₀ heterocycloalkyl or substituted or unsubstituted heteroaryl;

each R ⁷ in -NHC(=O)R ⁷ , -C(=O) ₂ HR ⁷ , -NHS(=O) ₂ R ⁷ , -S(=O) ₂ NHR ⁷ ; -NHC(=O) ₂ HR ⁷ , -NHS(=O) ₂ NHR ⁷ , -(C ₁ -C ₃ alkyl)-NHC(=O)R ⁷ , -(C ₁ -C ₃ alkyl)-C( =O) ₂ HR ⁷ , -(C ₁ -C ₃ alkyl)-NTS(=O)2NHR ⁷ , -(C ₁ -C ₃ alkyl)-S(=O) ₂ NHR ⁷ ; -(C ₁ -C ₃ alkyl)-NHC(=O) ₂ HR ⁷ , -(C ₁ -C ₃ alkyl)-NHS(=O) ₂ NHR ⁷ independently selected from C ₁ -C ₆ alkyl, C ₁ - C ₆ haloalkyl, C ₁ -C ₆ heteroalkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₂ -C ₁₀ heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl), -C ₁ -C ₆ alkyl- (substituted or unsubstituted C ₂ -C ₁₀ heterocycloalkyl, -C ₁ -C ₆ alkyl- (substituted or unsubstituted aryl), -C ₁ -C ₆ alkyl-(substituted or unsubstituted heteroaryl), -(CH ₂ ) _p -CH (substituted or unsubstituted aryl) ₂ , -(CH ₂ ) _p -CH (substituted or unsubstituted heteroaryl) ₂ , -(CH ₂ ) _p -CH(substituted or unsubstituted aryl)(substituted or unsubstituted heteroaryl), -(substituted or unsubstituted aryl)-(substituted or unsubstituted aryl), -(substituted or unsubstituted aryl)-(substituted or unsubstituted heteroaryl), -(substituted or unsubstituted heteroaryl)-(substituted or unsubstituted aryl) or -(substituted or unsubstituted heteroaryl)-(substituted or unsubstituted heteroaryl);

p in R ⁷ is selected from 0, 1 or 2;

R ^8a , R ^8b , R ^8c , R ^8d , R ^8e and R ^8f in C(R ^8a )(R ^8b ), C(R ^8c )(R ^8d ) and C(R ^8e )(R ^8f ) are independently selected from H, C ₁ -C ₆ alkyl, C ₁ -C ₆ fluoroalkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ heteroalkyl and substituted or unsubstituted aryl; or:

R ^8a , R ^8d , R ^8e and R ^8f in C(R ^8a )(R ^8b ), C(R ^8c )(R ^8d ) and C(R ^8e )(R ^8f ) are as defined above, and R ^8b and R ^8c together form a bond;

or:

R ^8a , R ^8b , R ^8d and R ^8f in C(R ^8a )(R ^8b ), C(R ^8c )(R ^8d ) and C(R ^8e )(R ^8f ) are such as

as defined above, and R ^8c and R ^8e together form a bond;

or:

R ^8a , R ^8d , R ^8e and R ^8f in C(R ^8a )(R ^8b ), C(R ^8c )(R ^8d ) and C(R ^8e )(R ^8f ) are as defined above, and R ^8b and R ^8c together with the atoms to which they are attached form a substituted or unsubstituted fused 5-7 membered saturated or partially saturated carbocyclic ring or a heterocyclic ring containing 1-3 heteroatoms selected from S, O and N, substituted or unsubstituted fused 5 -10 membered aryl ring or substituted or unsubstituted fused 5-10 membered heteroaryl ring containing 1-3 heteroatoms selected from S, O and N;

or:

R ^8a , R ^8b , R ^8d and R ^8f in C(R ^8a )(R ^8b ), C(R ^8c )(R ^8d ) and C(R ^8e )(R ^8f ) are as defined above, and R ^8c and R ^8e together with the atoms to which they are attached form a substituted or unsubstituted fused 5-7 membered saturated or partially saturated carbocyclic ring or a heterocyclic ring containing 1-3 heteroatoms selected from S, O and N, substituted or unsubstituted fused 5 -10 membered aryl ring or substituted or unsubstituted fused 5-10 membered heteroaryl ring containing 1-3 heteroatoms selected from S, O and N; or:

R ^8c , R ^8d , R ^8e and R ^8f in C(R ^8c )(R ^8d ) and C(R ^8e )(R ^8f ) are as defined above, and R ^8a and R ^8b together with the atoms to which they attached, forming a substituted or unsubstituted, saturated or partially saturated 3-7 membered spirocycle or heterospirocycle containing 1-3 heteroatoms selected from S, O and N; or:

R ^8a , R ^8b , R ^8e and R ^8f in C(R ^8a )(R ^8b ) and C(R ^8e )(R ^8f ) are as defined above, and R ^8c and R ^8d together with the atoms to which they attached, forming a substituted or unsubstituted, saturated or partially saturated 3-7 membered spirocycle or heterospirocycle containing 1-3 heteroatoms selected from S, O and N; or:

R ^8a , R ^8b , R ^8c and R ^8d in C(R ^8a )(R ^8b ) and C(R ^8c )(R ^8d ) are as defined above, and R ^8e and R ^8f together with the atoms to which they attached, forming a substituted or unsubstituted, saturated or partially saturated 3-7 membered spirocycle or heterospirocycle containing 1-3 heteroatoms selected from S, O and N; or:

wherein each substituted alkyl, heteroalkyl, fused ring, spirocycle, heterospirocycle, cycloalkyl, heterocycloalkyl, aryl or heteroaryl is substituted with 1 -3 R ⁹ ; And

each R ⁹ in R ^8a , R ^8b , R ^8c , R ^8d , R ^8e n R ^8f is independently selected from halogen, -OH -SH, (C=O), CN, C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, Q-C4 alkoxy, C ₁ -C ₄ fluoroalkoxy, -NH ₂ , -NH(C ₁ -C ₄ alkyl), -NH(C ₁ -C ₄ alkyl) ₂ , - C(=O)OH, -C(=O) ₂ H ₂ , -C(=O)C ₁ -C ₃ alkyl, -S(=O) ₂ CH ₃ , -NH(C ₁ -C ₄ alkyl)-OH, -NH(C ₁ -C ₄ alkyl)-O-(C-C ₄ alkyl), -O(C ₁ -C ₄ alkyl)-NH ₂ ; -O(C ₁ -C ₄ alkyl)-NH-(C ₁ -C ₄ alkyl) and -O(C ₁ -C ₄ alkyl)-N-(C ₁ -C ₄ alkyl) ₂ , or two R ⁹ together with the atoms to which they are attached, form a methylenedioxy or ethylenedioxy ring, substituted or unsubstituted with halogen, -OH or C ₁ -C ₃ alkyl.

[00110] In any of the compounds described herein, the ILM may have the structure of formulas (XLV), (XLVI) or (XLVII), which are derivatives of the TAP ligands described in Vamos, M., et al., Expedient synthesis of highly potent antagonists of inhibitor of apoptosis proteins (IAPs) with unique selectivity for ML-IAP, ACS Chem. Biol., 8(4), 725-32 (2013), or its non-natural mimetic:

Where:

R ^2f R ³ and R ⁴ of formula (XLV) are independently selected from H or ME;

X of formula (XLV) is independently selected from O or S; And

R ¹ formula (XLV) selected from:

[00111] In a specific embodiment, the ELM has a structure according to formula (XLVIII):

where R ³ and R ⁴ of formula (XLVIH) are independently selected from H or ME;

is a 5-membered heterocycle selected from:

[00112] In a particular embodiment formula XLVIII) is

[00113] In a specific embodiment, the ILM has the structure and is attached to the linker group L as shown below:

[00114] In a specific embodiment, the ILM has a structure in accordance with formula (XLIX), (L) or (LI):

Where:

R ³ of formula (XLIX), (L) or (LI) are independently selected from H or ME;

is a 5-membered heterocycle selected from:

And

L formulas (XLIX), (L) or (LI) selected from:

[00115] In a specific embodiment L of formula (XLIX), (L) or (LI)

[00116] In a specific embodiment, the ILM has a structure in accordance with formula (LII):

[00117] In a specific embodiment, the ILM according to formula (LII) is chemically bonded to the linker group L in the region

designated with and as below:

[00118] In any of the compounds described herein, the ILM may have a structure of formulas (LIII) or (LIV), which are based on the IAP ligands described in Hennessy, EJ, et al., Discovery of aminopiperidine-based Smac mimetics as JAP antagonists , Bioorg. Med Chem. Lett., 22(4), 1960-4 (2012) or its non-natural mimetic:

Where:

R ¹ of formula (LIII) n (LIV) selected from:

R ² of formula (LIII) n (LIV) is selected from H or Me;

R ³ of formula (LIII) n (LIV) selected from:

X is selected from H, halogen, methyl, methoxy, hydroxy, nitrogen or trifluoromethyl.

[00119] In any of the compounds described herein, the ILM may have the structure and be chemically associated with a linker as shown in formula (LV) or (LVI), or a non-natural mimetic thereof:

[00120] In any of the compounds described herein, the ILM may have a structure of formula (LVII), which is based on the LAP ligands described in Cohen, F, et al., Orally bioavailable antagonists of inhibitor of apoptosis proteins based on an azabicyclooctane scaffold, J. Med. Chem., 52(6), 1723-30 (2009), or its non-natural mimetic:

Where:

R ¹ formulas (LVII) selected from:

X selected from H, fluorine, methyl or methoxy.

[00121] In a specific embodiment, the ILM is represented by the following structure:

[00122] In a specific embodiment, the ILM is selected from the group of, in the composition of, and with a chemical bond between the ILM and the linker group L, represented by:

[00123] In any of the compounds described herein, the ILM is selected from the group consisting of the structures below that are based on the LAP ligands described in Asano, M, et al., Design, sterioselective synthesis, and biological evaluation of novel tri-cyclic compounds as inhibitor of apoptosis proteins (JAP) antagonists, Bioorg. Med. Chem., 21(18): 5725-37 (2013), or its non-natural mimetic:

[00124] In a specific embodiment, the ILM is selected from the group of, in the composition of, and with a chemical bond between the ILM and the linker group L, represented by:

[00125] In any of the compounds described herein, the ILM may have a structure of formula (LVIII), which is based on the TAP ligands described in Asano, M, et al., Design, sterioselective synthesis, and biological evaluation of novel tri-cyclic compounds as inhibitor of apoptosis proteins (LAP) antagonists, Bioorg. Med Chem., 21(18): 5725-37 (2013), or its non-natural mimetic:

where X of formula (LVIII) represents one or two substituents independently selected from H, halogen or cyano.

[00126] In any of the compounds described herein, ILM may have the structure and be chemically associated with the linker group L, as shown in formula (LIX) or (LX), or a non-natural mimetic thereof:

where X of formula (LIX) and (LX) represents one or two substituents independently selected from H, halogen or cyano; and L of formulas (LIX) and (LX) represents a linker group as described herein.

[00127] In any of the compounds described herein, the ILM may have a structure of formula (LXI), which is based on the IAP ligands described in Ardecky, RJ, et al., Design, sysnthesis and evaluation of inhibitor of apoptosis (LAP) antagonists that are highly selective for the BIR2 domain of XIAP, Bioorg. Med Chem., 23(14): 4253-7 (2013), or its non-natural mimetic:

Where:

formula (LXI) is a natural or unnatural amino acid; And

R ² formula (LXI) selected from:

[00128] In any of the compounds described herein, ILM may have the structure and be chemically associated with the linker group L as shown in formula (LXH) or (LLXIH), or a non-natural mimetic thereof:

formula (LXI) is a natural or unnatural amino acid; And

L of formula (LXI) is a linker group as described herein.

[00129] In any of the compounds described herein, the ILM may have a structure selected from the group consisting of ligand-based LAPs described in Wang, J, et al., Discovery of novel second mitochondrial-derived activator of caspase mimetics as selective inhibitor or apoptosis protein inhibitors, J. Pharmacol. Exp.Tlier., 349(2): 319-29 (2014), or its non-natural mimetic:

[00130] In any of the compounds described herein, the ELM has a structure according to formula (LXIX), which is based on the LAP ligands described in Hird, AW, et al., Structure-based design and synthesis of tricyclic LAP (Inhibitors' of Apoptosis Proteins) inhibitors, Bio org. Med Chem. Lett., 24(7): 1820-4 (2014), or its non-natural mimetic:

where R of formula LLX is selected from the group consisting of:

R1 in selected from H or Me;

R2 in selected from alkyl or cycloalkyl;

X in represents 1-2 substituents independently selected from halogen, hydroxy, methoxy, nitrogen and trifluoromethyl

Z in represents O or NH;

No in is a mono- or fused bicyclic heteroaryl; And

formula (LLX) represents an optional double bond.

[00131] In a specific embodiment, the ILM compound has a chemical structure represented by:

[00132] In a specific embodiment, the ELM compound has a chemical structure selected from the group consisting of:

[00133] The term "independently" is used herein to indicate that a variable that is applied independently changes regardless of application to application.

[00134] The term "alkyl" shall mean in its context a linear, branched or cyclic fully saturated hydrocarbon radical or alkyl group, preferably a C ₁ -C ₁₀ , more preferably a C ₁ -C ₆ , alternatively a C ₁ -C ₃ alkyl group, which may optionally be substituted. Examples of alkyl groups are methyl, ethyl, n-butyl, sec-butyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, isopropyl, 2-methylpropyl, cyclopropyl, cyclopropylmethyl, cyclobutyl, cyclopentyl, cyclopentylethyl, cyclohexylethyl and cyclohexyl, among others. In some embodiments, the alkyl group is terminated with a halogen group (At, Br, Cg, F, or I). In some preferred embodiments, the compounds of the invention may be used to covalently bind to dehalogenase enzymes. These compounds typically contain a side chain (often linked through a polyethylene glycol group) that is terminated by an alkyl group that has a halogen substituent (often chlorine or bromine) at its distal end, resulting in covalent binding of the compound containing such a moiety to the protein.

[00135] The term "alkenyl" refers to linear, branched or cyclic C ₂ -C ₁₀ (preferably C ₂ -C ₆ ) hydrocarbon radicals containing at least one C=C bond.

[00136] The term "alkynyl" refers to linear, branched or cyclic C ₂ -C ₁₀ (preferably C ₂ -C ₆ ) hydrocarbon radicals containing at least one C=C bond.

[00137] The term "alkylene" as used refers to the group - (CH ₂ ) _n - (n is an integer, typically 0 to 6), which may be optionally substituted. When substituted, the alkylene group is preferably substituted on one or more methylene groups with a C ₁ -C ₆ alkyl group (including a cyclopropyl group or tert-butyl group), but may also be substituted with one or more halogen groups, preferably 1 to 3 halogen groups or one or two hydroxyl groups, O-(C ₁ -C ₆ alkyl) groups or amino acid side chains as otherwise disclosed herein. In some embodiments, the alkylene group may be replaced by a urethane or alkoxy group (or other group) that is further substituted by a polyethylene glycol chain (1 to 10, preferably 1 to 6, often 1 to 4 ethylene glycol units) that is replaced by (preferably but not exclusively at the distal end of the polyethylene glycol chain) an alkyl chain substituted by one halogen group, preferably a chlorine group. In other embodiments, the alkylene (often methylene) group may be replaced by an amino acid side chain group, such as a natural or unnatural amino acid side chain group, e.g., alanine, p-alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine , glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan or tyrosine.

[00138] The term "unsubstituted" shall mean substituted only with hydrogen atoms. The range of carbon atoms that includes C ₀ means that carbon is missing and is replaced by H. Thus, the range of carbon atoms that is C ₀ -C ₆ includes carbon atoms 1, 2, 3, 4, 5 and 6, and for _C0 , H replaces carbon.

[00139] The term "substituted" or "optionally substituted" shall mean independently (i.e., when many substitutions occur, each substituent is independent of the other substituent) one or more substituents (up to five substituents independently, preferably up to three substituents, often 1 or 2 substituents on a moiety in a compound as described herein and may include substituents that themselves may be further substituted) at a carbon (or nitrogen) position anywhere on the molecule in context, and include as substituents hydroxyl, thiol, carboxyl, cyano (C =N), nitro (NO ₂ ), halogen (preferably 1, 2 or 3 halogen, especially on an alkyl, especially on a methyl group such as trifluoromethyl), alkyl group (preferably C ₁ -C ₁₀ , more preferably C _{1 -C6} ), aryl (preferably phenyl and substituted phenyl, such as benzyl or benzoyl), alkoxy (preferably _C1 - _C6 alkyl or aryl, including phenyl and substituted phenyl), thioether ( _C1 - _C6 alkyl or aryl) , acyl (preferably C ₁ -C ₆ acyl), ester or thioester (preferably C ₁ -C ₆ alkyl or aryl), including an alkylene ester (so that the attachment occurs at the alkylene group rather than at the ester group, which preferably substituted with a C ₁ -C ₆ alkyl or aryl group), preferably a C ₁ -C ₆ alkyl or aryl, halogen (preferably F or Cl), amine (including a five- or six-membered cyclic alkyleneamine, additionally including a C ₁ -C ₆ alkylamine or a C ₁ -C ₆ dialkylamine whose alkyl groups may be substituted by one or two hydroxyl groups) or an optionally substituted -N(C ₀ -C ₆ alkyl)C(O)(OC ₁ -C ₆ alkyl) group (which may optionally be substituted with a polyethylene glycol chain, to which is further bonded an alkyl group containing one halogen, preferably a chlorine substituent), hydrazine, amido, which is preferably substituted with one or two C ₁ -C ₆ alkyl groups (including a carboxamide, which is optionally substituted with one or two C ₁ - With ₆ alkyl groups), an alkanol (preferably a C ₁ -C ₆ alkyl or aryl) or an alkanoic acid (preferably a C ₁ -C ₆ alkyl or aryl). Substituents in accordance with the present invention may include, for example, SiR ₁ R ₂ R ₃ groups, where each of R ₁ and R ₂ is as described herein, and R ₃ represents H or a C ₁ -C ₆ alkyl group preferably R ₁ , R ₂ , R ₃ in this context represents a C ₁ -C ₃ alkyl group (including an isopropyl or tert-butyl group). Each of the groups described above may be linked directly to the substituted moiety or, alternatively, the substituent may be linked to the substituted moiety (preferably in the case of an aryl or heteraryl moiety) via an optionally substituted -(CH ₂ ) _m - or, alternatively, an optionally substituted -( OCH ₂ ) _m , -(OSH ₂ CH ₂ ) _m or -(CH ₂ CH ₂ O) _m is a group that may be substituted by one or more of the substituents described above. Alkylene groups -(CH ₂ ) _m or -(CH ₂ ) _m - groups or other chains, such as ethylene glycol chains, as indicated above, can be substituted anywhere on the chain. Preferred substituents on alkylene groups include halogen or C ₁ -C ₆ (preferably C ₁ -C ₃ ) alkyl groups, which may be optionally substituted with one or two hydroxyl groups, one or two ether groups (O-C ₁ -C ₆ groups) , up to three halogen groups (preferably F), or an amino acid side chain as described herein, and an optionally substituted amide (preferably a carboxamide substituted as described above) or urethane groups (often with one or two _C0 - _C6 alkyl substituents, wherein the group(s) may be further substituted(s). In certain embodiments, the alkylene group (often one methylene group) is substituted with one or two optionally substituted C ₁ -C ₆ alkyl groups, preferably a C ₁ -C ₄ alkyl group, most often a methyl or O-methyl group or an amino acid side chain, as otherwise described in this document. In the present invention, a moiety in the molecule may optionally be substituted with up to five substituents, preferably up to three substituents. Most often, as used herein, substituted moieties are substituted with one or two substituents.

[00140] The term "substituted" (each substituent is independent of any other substituent) shall also mean, in its context of use, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, halogen, amido, carboxamido, sulfone, including sulfonamide, keto , carboxy, C ₁ -C ₆ ester (hydroxy ester or carbonyl ester), C ₁ -C ₆ keto, urethane -OC(O)-NR ₁ R ₂ or -N(R ₁ )-C(O)-OR ₁ , nitrogen, piano and amine (particularly including C ₁ -C ₆ alkylene-R ₁ R ₂ , mono- or di-C ₁ -C ₆ alkyl-substituted amines, which may be optionally substituted with one or two hydroxyl groups). Each of these groups contains, unless otherwise indicated, in context from 1 to 6 carbon atoms. In certain embodiments, preferred substituents will include, for example, -NH-, -NHC(O)-, -O-,=O, -( _CH2 ) _m (as used herein, 1, 2, 3 , 4, 5 or 6), -S-, -S(O)-, SO ₂ - or -NH-C(O)-NH-, -(CH ₂ ) _n OH, -(CH ₂ ) ₂ SH, -(CH ₂ ) _n COOH, C ₁ -C ₆ alkyl, -(CH ₂ ) _n -O-(C ₁ -C ₆ alkyl), -(CH ₂ ) _n C(O)-(C ₁ -C ₆ alkyl), -(CH ₂ ) _n OC(O)-(C ₁ -C ₆ alkyl), -(CH ₂ ) _n C(O)O-(C ₁ -C ₆ alkyl), -(CH ₂ ) _n NHC(O)-R ₁ , -(CH ₂ ) ₂ C(O)-NR ₁ R ₂ , -(OSH ₂ ) _n OH,. (CH ₂ O) _n COOH, C ₁ -C ₆ alkyl, -(OCH ₂ ) _n O-(C ₁ -C ₆ alkyl), -(CH ₂ O)nC(O)-(C ₁ -C ₆ alkyl ), -(OCH ₂ ) ₂ NHC(O)-R ¹ , -(CH ₂ O) ₂ C(O)-NR ₁ R ₂ , -S(O) ₂ -Rs, -S(O)-R _s (R _s represents a C ₁ -C ₆ alkyl or -(CH ₂ ) _m -NR ₁ R ₂ group), NO ₂ , CN or halogen (F, Cl, Br, I, preferably F or Cl), depending on context of using the substitute. Each R ₁ and R ₂ represents, within the context, H or a C ₁ -C ₆ alkyl group (which may optionally be substituted by one or two hydroxyl groups or a maximum of three halogen groups, preferably fluorine). The term “substituted” shall also mean, in the chemical context of the particular compound and substituent used, an optionally substituted aryl or heteroaryl group, or an optionally substituted heterocyclic group, as otherwise described herein. Alkylene groups may also be substituted as described herein, preferably with optionally substituted C ₁ -C ₆ alkyl groups (preferably methyl, ethyl or hydroxymethyl or hydroxyethyl, thereby providing a chiral center), a side chain of an amino acid group as otherwise described herein document, an amido group as described above or a urethane group OC(O)-NR ₁ R ₂ wherein R ₁ and R ₂ have other meanings as described herein, although many other groups may also be used as substituents. The various optionally substituted moieties may be substituted with 3 or more substituents, preferably no more than 3 substituents and preferably 1 or 2 substituents. It should be noted that in cases where a substitution is required in a compound at a particular position of the molecule (primarily due to valency) but the substitution is not specified, then that substituent is construed or understood to be H unless the context of the substitution suggests otherwise.

[00141] The term "aryl" or "aromatic" as used herein refers to a substituted (as otherwise described herein) or unsubstituted monovalent aromatic radical having a single ring (e.g., benzene, phenyl, benzyl) or fused rings (e.g., naphthyl, anthracenyl, phenanthrenyl, etc.) and may be linked to a compound as described herein at any available stable position on the ring(s) or as otherwise indicated in the chemical structure provided. Other examples of aryl groups, in context, may include heterocyclic aromatic ring systems, "heteroaryl" groups having one or more nitrogen, oxygen or sulfur atoms on the ring (moncyclic), such as imidazole, furyl, pyrrole, furanyl, thiene, thiazole, pyridine, pyrimidine, pyrazine, triazole, oxazole or fused ring systems such as indole, quinoline, indolizine, azaindolizine, benzofurazan, etc., among others, which may be optionally substituted as described above. Among the heteroaryl groups that may be mentioned are nitrogen-containing heteroaryl groups such as pyrrole, pyridine, pyridone, pyridazine, pyrimidine, pyrazine, pyrazole, imidazole, triazole, triazine, tetrazole, indole, isoindole, indolysine, azaindolysine, purine, indazole, quinoline, dihydroquinoline, tetrahydroquinoline, isoquinoline, dihydroisoquinoline, tetrahydroisoquinoline, quinolysine, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, imidazopyridine, imidazotriazine, pyrazinopyridazine, acridine, phenanthridine, carbazole, carbazoline, pyrimi dine, phenanthroline, phenacene, oxadiazole, benzimidazole, pyrrolopyridine, pyrrolopyrimidine and pyridopyrimidine; sulfur-containing aromatic heterocycles such as thiophene and benzothiophene; oxygen-containing aromatic heterocycles such as furan, pyran, cyclopentapyran, benzofuran and isobenzofuran; and aromatic heterocycles containing 2 or more heteroatoms selected from nitrogen, sulfur and oxygen, such as thiazole, thiadizole, isothiazole, benzoxazole, benzothiazole, benzothiadiazole, phenothiazine, isoxazole, furazan, phenoxazine, pyrazoloxazole, imidazothiazole, thienofuran, furopyrrole, pyridoxazine , furopyridine, furopyrimidine, thienopyrimidine and oxazole, among others, all of which may be optionally substituted.

[00142] The term “substituted aryl” refers to an aromatic carbocyclic group consisting of at least one aromatic ring or more fused rings, at least one of which is aromatic, wherein the ring(s) is substituted with one or more substituents. For example, the aryl group may contain substituent(s) selected from: -( _CH2 ) _nOH , -( _CH2 ) _n -O-( _C1 - _C6 )alkyl, -( _CH2 ) _n -O-(CH ₂ ) _n -(C ₁ -C ₆ ) alkyl, -(CH ₂ ) _n -C(O)(C ₀ -C ₆ ) alkyl, -(CH ₂ ) _n -C(O) O(C ₀ -C ₆ )alkyl, -(CH ₂ ) _n -OC(O)(C ₀ -C ₆ )alkyl, amine, mono- or di-(C ₁ -C ₆ alkyl) amine, where the alkyl group amine is optionally substituted with 1 or 2 hydroxyl groups or up to three halogen groups (preferably F, Cl), OH, COOH, C ₁ -C ₆ alkyl, preferably CH ₃ , CF ₃ , OMe, OCF ₃ , NO ₂ or CN groups (each of which may be substituted at the ortho, meta and/or para positions of the phenyl ring, preferably para), an optionally substituted phenyl group (the phenyl group itself is preferably linked/attached to the PTM group, including the ULM group, via a linker group), and/or at least one of F, Cl, OH, COOH, CH ₃ , CF ₃ , OMe, OCF ₃ , NO ₂ or CN groups (at the ortho, meta and/or para positions of the phenyl ring, preferably para -), a naphthyl group, which may be optionally substituted, an optionally substituted heteroaryl, preferably an optionally substituted isoxazole, including a methyl-substituted isoxazole, an optionally substituted oxazole, including a methyl-substituted oxazole, an optionally substituted thiazole, including a methyl-substituted thiazole, an optionally substituted isothiazole, including a methyl-substituted isothiazole, optional substituted pyrrole, including methyl-substituted pyrrole, optionally substituted imidazole, including methylimidazole, optionally substituted benzimidazole or methoxybenzylimidazole, optionally substituted oximidazole or methyloximidazole, optionally substituted diazole group, including methyldiazole group, optionally substituted triazole group, including methyl-substituted tria an ash group, an optionally substituted pyridine group, including a halogen (preferably F) or methyl substituted pyridine group, or oxapyridine group (where the pyridine group is linked to the phenyl group via an oxygen), optionally substituted furan, optionally substituted benzofuran, optionally substituted dihydrobenzofuran, optionally substituted indole, indolizine or azaindolizine (2 , 3 or 4-azaindolizine), optionally substituted quinoline and combinations thereof.

[00143] "Carboxyl" means a -C(O)OR group, where R is hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl or substituted heteroaryl, while these general substituents have meanings that are identical to the definitions of the corresponding groups, defined in this document.

[00144] The term “heteroaryl” or “hetaryl” may mean, but is in no way limited to, optionally substituted quinoline (which may be attached to a pharmacophore or substituted on any carbon atom on the quinoline ring), optionally substituted indole (including dihydroindole), optionally substituted indolizine, optionally substituted azaindolizine (2, 3 or 4-azaindolizine), optionally substituted benzimidazole, benzodiazole, benzoxofuran, optionally substituted imidazole, optionally substituted isoxazole, optionally substituted oxazole (preferably methyl substituted), optionally substituted diazole, optionally substituted triazole, tetra angry , optionally substituted benzofuran, optionally substituted thiophene, optionally substituted thiazole (preferably substituted with methyl and/or thiol), optionally substituted isothiazole, optionally substituted triazole (preferably 1,2,3-triazole substituted with a methyl group, triisopropylsilyl group, optionally substituted -( CH ₂ ) _m OC ₁ -C ₆ alkyl group or an optionally substituted -(CH ₂ ) _m C(O)-OC ₁ -C ₆ alkyl group), optionally substituted pyridine (2-, 3 or 4-pyridine) or a group in according to chemical structure:

Where:

S ^c represents CHR ^SS , NR ^URE or O;

R ^HET is H, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), optionally substituted with O( C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or an optionally substituted acetylene group -C≡C-Ra, where Ra represents H or a C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl);

R ^SS is H, CN, NO ₂ , halogen (preferably F or O), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups), optionally substituted O-(C ₁ - C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted -C(O)(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups);

R ^URE is H, C ₁ -C ₆ alkyl (preferably H or C ₁ -C ₃ alkyl) or -C(O)(C ₁ -C ₆ alkyl), each of which is optionally substituted with one or two hydroxyl groups or up to three halogens, preferably fluorine groups, or an optionally substituted heterocycle, for example piperidine, morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, piperidine, piperazine, each of which is optionally substituted, and

Y ^C is N or CR ^YC, where R ^YC is H, OH, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), an optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or an optionally substituted acetylene group -C≡CR _a where R _a is H or a C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl).

[00145] The terms "aralkyl" and "heteroarylalkyl" refer to groups that include both aryl and, respectively, heteroaryl, as well as alkyl and/or heteroalkyl and/or carbocyclic and/or heterocycloalkyl ring systems according to the following above definitions.

[00146] As used herein, the term “arylalkyl” refers to an aryl group, as defined above, attached to an alkyl group, as defined above. The arylalkyl group is attached to the parent moiety through an alkyl group, in which the alkyl group contains from one to six carbon atoms. The aryl group in the arylalkyl group may be substituted as defined above.

[00147] The term “heterocycle” refers to a cyclic group that contains at least one heteroatom, such as N, O, or S, and may be aromatic (heteroaryl) or non-aromatic. Thus, heteroaryl moieties fall under the definition of a heterocycle depending on the context of its use. Typical heteroaryl groups are described above.

[00148] Representative heterocycles include: azetidinyl, benzimidazolyl, 1,4-benzodioxanyl, 1,3-benzodioxolyl, benzoxazolyl, benzothiazolyl, benzothienyl, dihydroimidazolyl, dihydropyranyl, dihydrofuranyl, dioxanyl, dioxolanil, ethylene urea, 1,3-dioxolane, 1,3 -dioxane, 1,4-dioxane, furyl, homopiperidinyl, imidazolyl, imidazolinyl, imidazolidinyl, indolinyl, indolyl, isoquinolinyl, isothiazolidinyl, isothiazolyl, isoxazolidinyl, isoxazolyl, morpholinyl, naphthyridinyl, oxazolidinyl, oxazolyl, pyridone, 2-pyrrolidone, pyri din, piperazinil . , oxetanil, oxathiolanil, Tian and others.

[00149] Heterocyclic groups may optionally be substituted by a member selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido - SO-alkyl , -SO-substituted alkyl, -SOaryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, oxo (=O) and -SO2-heteroaryl. Such heterocyclic groups may have one ring or several fused rings. Examples of nitrogen heterocycles and heteroaryls include, but are not limited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolysine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, morpholilo, piperidinyl, tetrahydrofuranyl and the like, as well as heterocycles containing N- alkoxy-nitrogen. The term "heterocyclic" also includes bicyclic groups in which any of the heterocyclic rings are fused to a benzene ring or a cyclohexane ring or other heterocyclic ring (eg, indolyl, quinolyl, isoquinolyl, tetrahydroquinolyl, etc.).

[00150] The term “cycloalkyl” may mean, but is in no way limited to, monovalent groups derived from monocyclic or polycyclic alkyl groups or cycloalkanes as defined herein, for example, saturated monocyclic hydrocarbon groups containing from three to twenty carbon atoms in the ring, including, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. The term "substituted cycloalkyl" may mean, but is in no way limited to, a monocyclic or polycyclic alkyl group substituted with one or more substituents, for example amino, halogen, alkyl, substituted alkyl, carbiloxy, carbyl mercapto, aryl, nitro, mercapto or sulfo, then how these common substituent groups have meanings that are identical to the definitions of the corresponding groups as defined in these legends.

[00151] “Heterocycloalkyl” refers to a monocyclic or polycyclic alkyl group in which at least one ring carbon atom of its cyclic structure is replaced by a heteroatom selected from the group consisting of N, O, S or P. “Substituted heterocycloalkyl” refers to a monocyclic or a polycyclic alkyl group in which at least one ring carbon atom of its cyclic structure is replaced by a heteroatom selected from the group consisting of N, O, S or P, and the group contains one or more substituents selected from the group consisting of halogen, alkyl, substituted alkyl, carbyloxy, carbylmercapto, aryl, nitro, mercapto or sulfo, whereas these general substituent groups have meanings that are identical to the definitions of the corresponding groups as defined in these legends.

[00152] The term “hydrocarbyl” means a compound that contains carbon and hydrogen and which may be fully saturated, partially unsaturated or aromatic and includes aryl groups, alkyl groups, alkenyl groups and alkynyl groups.

[00153] The term “independently” is used herein to indicate that a variable that is applied independently changes regardless of application to application.

[00154] The term "lower alkyl" refers to methyl, ethyl or propyl.

[00155] The term "lower alkoxy" refers to methoxy, ethoxy or propoxy.

[00156] In any of the embodiments described herein, W, X, Y, Z, G, G', R, R', R'', Q1-Q4, A, and Rn may be independently covalently linked to the linker and/or a linker to which one or more PTM, ULM, ILM or ILM groups are attached.

[00157] Typical MLM

[00158] In certain additional embodiments, the MLM of the bifunctional compound includes chemical moieties such as substituted imidazolines, substituted spiroindolinones, substituted pyrrolidines, substituted piperidinones, substituted morpholinones, substituted pyrrolopyrimidines, substituted imidazolopyridines, substituted thiazoloimidazolinones, and substituted isochs nolinones.

[00159] In additional embodiments, the MLM contains the above core structures with adjacent bis-aryl substitutions positioned as cis or trans configurations.

[00160] In still further embodiments, the MLM comprises a portion of structural elements such as those in RG7112, RG7388, SAR405838, AMG-232, AM-7209, DS-5272, MK-8242, and NVP-CGM-097, as well as analogs thereof or derivatives.

[00161] In some preferred embodiments, MLM is a derivative of a substituted imidazoline represented by formula (A-1), or a thiazoloimidazoline represented by formula (A-2), or a spiroindolinone represented by formula (A-3), or a pyrrolidine represented by formula (A-4), or piperidinone/morphlinone represented by formula (A-5), or isoquinolinone represented by formula (A-6), or pyrollopyrimidine/imidazolopyridine represented by formula (A-7), or pyrrolopyrrolidinone/imidazolopyrrolidinone represented by formula (A-8).

where in the above formula (A-1) - formula (A-8),

X of formula (A-1) - formula (A-8) is selected from the group consisting of carbon, oxygen, sulfur, sulfoxide, sulfone and N-Ra;

Ra represents independently H or an alkyl group of 1-6 carbon atoms;

Y and Z of formula (A-1) to formula (A-8) are independently carbon or nitrogen;

A, A' and A'' of formula (A-1) - formula (A-8) are independently selected from C, N, O or S, may also be one or two atoms forming a fused bicyclic ring or 6,5- and a 5,5-fused aromatic bicyclic group;

R1, R2 of formula (A-1) to formula (A-8) are independently selected from the group consisting of an aryl or heteroaryl group, a heteroaryl group having one or two heteroatoms independently selected from sulfur or nitrogen, wherein the aryl or heteroaryl group may be monocyclic or bicyclic, or unsubstituted or substituted with one to three substituents independently selected from the group consisting of: halogen, -CN, C1-C6 alkyl group, C3-C6 cycloalkyl, -OH, 1-6 carbon alkoxy, fluoro-substituted 1-6 carbon alkoxy, 1-6 carbon sulfoxide, 1-6 carbon sulfone, 2-6 carbon ketone, 2-6 carbon amides and 2-6 carbon dialkylamine;

R ₃ , R ₄ of formula (A-1) - formula (A-8) are independently selected from the group consisting of H, methyl and C1 - C6 alkyl;

R ₅ of formula (A-1) to formula (A-8) is selected from the group consisting of an aryl or heteroaryl group, a heteroaryl group having one or two heteroatoms independently selected from sulfur or nitrogen, wherein the aryl or heteroaryl group may be monocyclic or bicyclic, or unsubstituted or substituted with one to three substituents independently selected from the group consisting of:

halogen, -CN, C1 - C6 alkyl group, C3-C6 cycloalkyl, -OH, alkoxy with 1-6 carbons, fluorinated alkoxy with 1-6 carbons, sulfoxide with 1-6 carbons, sulfone with 1-6 carbons, ketone with 2-6 carbons, amides with 2-6 carbons, dialkylamine with 2-6 carbons, alkyl ether (C2 - C6), alkyl ketone (C3-C6), morpholinyl, alkyl ester (C3-C6), alkyl cyanide (C3- C6);

R ⁶ of formula (A-1) - formula (A-8) is H or -C(=O)R ^b , where R ^{b of} formula (A-1) - formula (A-8) is selected from the group consisting of alkyl, cycloalkyl, mono-, di- or tri-substituted aryl or heteroaryl, 4-morpholinyl, 1-(3-oxopiperazunyl), 1-piperidinyl, 4- ^NRc- morpholinyl, 4- ^Rc -1-piperidinyl and 3-R ^c - 1-piperidinyl, where R ^c of formula (A-1) - formula (A-8) is selected from the group consisting of alkyl, fluoro-alkyl, cyano alkyl, hydroxyl-substituted alkyl, cycloalkyl, alkoxyalkyl, amidalkyl, alkylsulfone, alkylsulfoxide , alkylamide, aryl, heteroaryl, mono-, bis- and trisubstituted aryl or heteroaryl, CH2CH2R ^d and CH2CH2CH2R ^d , where

R ^d of formula (A-1) - formula (A-8) is selected from the group consisting of alkoxy, alkylsulfone, alkylsulfoxide, N-substituted carboxamide, -NHC(O)-alkyl, -NH-SO ₂ -alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl;

R ₇ of formula (Al) - formula (A-8) is selected from the group consisting of H, C1 - C6 alkyl, cyclic alkyl, fluoro-alkyl, cyano-substituted alkyl, 5- or 6-membered heteroaryl or 5- or 6-substituted aryl -membered heteroaryl or aryl;

R ₈ of formula (A-1) - formula (A-8) is selected from the group consisting of -R ^e -C(O)-R ^f , -R ^e -alkoxy, -R ^E -aloxy, -R ^e -heteroaryl and -R ^e -C(O)-R ^f -C(O)-R ^g where: R ^e of formula (A-1) to formula (A-8) is a 1-6 carbon alkylene or bond;

R ^f of formula (A-1) - formula (A-8) is a substituted 4-7 membered heterocycle;

R ^g of formula (A-1) - formula (A-8) is selected from the group consisting of aryl, heteroaryl, substituted aryl or heteroaryl, and a 4-7 membered heterocycle;

R ⁹ of formula (A-1) - formula (A-8) is selected from the group consisting of a mono-, bi- or tri-substituent on the fused bicyclic aromatic ring in formula (A-3), wherein the substituents are independently selected from the group consisting of halogen, alkene, alkyne, alkyl, unsubstituted or substituted with Cl or F;

R ₁₀ of formula (A-1) - formula (A-8) is selected from the group consisting of an aryl or heteroaryl group, where the heteroaryl group may contain one or two heteroatoms such as sulfur or nitrogen, the aryl or heteroaryl group may be monocyclic or A bicyclic, aryl or heteroaryl group may be unsubstituted or substituted with one to three substituents, including halogen, F, Cl, -CN, alkene, alkyne, C1 - C6 alkyl group, C1 - C6 cycloalkyl, -OH, 1-6 alkoxy carbons, fluorine substituted with 1-6 carbon alkoxy, 1-6 carbon sulfoxide, 1-6 carbon sulfone, 2-6 carbon ketone;

R ₁₁ of formula (A-1) - formula (A-8) is -C(O)-N(R ^h )(R ⁱ ), where R ^h and R ⁱ are selected from the group consisting of the following:

N; optionally substituted linear or branched C1 - C6 alkyl; alkoxy substituted alkyl; mono- and dihydroxy substituted alkyl (for example, C ₃ -C ₆ ), sulfone-substituted alkyl; optionally substituted aryl; optionally substituted heteroaryl; mono-, bis- or trisubstituted aryl or heteroaryl; phenyl-4-carboxylic acid; substituted phenyl-4-carboxylic acid, alkyl carboxylic acid; optionally substituted heteroaryl carboxylic acid; alkyl carboxylic acid; fluorine substituted alkyl carboxylic acid; optionally substituted cycloalkyl, 3-hydroxycyclobutane, 4-hydroxycyclohexane, aryl substituted cycloalkyl; heteroaryl substituted with cycloalkyl; or R ^h and R ⁱ taken together to form a ring;

R ₁₂ and R ₁₃ of formula (A-1) - formula (A-8) are independently selected from H, lower alkyl (C1 - C6), lower alkenyl (C2 - C6), lower alkynyl (C2 - C6), cycloalkyl (4 , 5 and 6 membered ring), substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, 5 and 6 membered aryl and heteroaryl, R ₁₂ and R ₁₃ can be linked to form a 5 and 6 membered ring with or without substitution on the ring ;

R ₁₄ of formula (A-1) - formula (A-8) is selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, substituted heterocycle, cycloalkyl, substituted cycloalkyl , cycloalkenyl and substituted cycloalkenyl;

R ₁₅ of formula (A-1) - formula (A-8) represents CN;

R ₁₆ of formula (A-1) - formula (A-8) is selected from the group consisting of C1-6 alkyl, C1-6 cycloalkyl, C2-6 alkenyl, C1-6 alkyl or C3-6 cycloalkyl with one or more hydrogens , substituted with fluorine, alkyl or cycloalkyl with one CH ₂ substituted by S(=O), -S or -S(=O) ₂ , alkyl or cycloalkyl with terminal CH ₃ substituted by 8(=O) ₂ N(alkyl)(alkyl) , -C(=O)N(alkyl)(alkyl), -N(alkyl)8(=O) ₂ (alkyl), -C(=O) ₂ (alkyl), O(alkyl), C1-6 alkyl or an alkylcycloalkyl with a hydron substituted by a hydroxyl group, a 3-7 membered cycloalkyl or heterocycloalkyl, optionally containing an -(C=0)- group, or a 5-6 membered aryl or heteroaryl group, the heterocycloalkyl or heteroaryl group of which may contain from one to three heteroatoms , independently selected from O, N or S, and the cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be unsubstituted or substituted with one or three substituents independently selected from halogen, C1-6 alkyl groups, hydroxylated C1-6 alkyl, C1-6 alkyl containing a thioether, ether, sulfone, sulfoxide, fluoroether or cyano group;

R ₁₇ of formula (A-1) - formula (A-8) is selected from the group consisting of (CH ₂ )nC(O) ₂ R ^k R ¹ , where R ^k and R ¹ are independently selected from H, C1-6 alkyl , hydroxylated C1-6 alkyl, C1-6 alkoxy alkyl, C1-6 alkyl with one or more hydrogens substituted by fluorine, C1-6 alkyl with one carbon substituted by S(O), S(O)(O), C1- 6 alkoxyalkyl with one or more hydrogens substituted with fluorine, C1-6 alkyl with a hydrogen substituted by a cyano group, 5 and 6 membered aryl or heteroaryl, alkylaryl with an alkyl group containing 1-6 carbons, and alkylheteroaryl with an alkyl group containing 1-6 carbons, where the aryl or heteroaryl group may be further substituted;

R ₁₈ of formula (A-1) to formula (A-8) is selected from the group consisting of substituted aryl, heteroaryl, alkyl, cycloalkyl, wherein the substitution is preferably -N(C1-4 alkyl)(cycloalkyl), -N (C1-4 alkyl)alkyl-cycloalkyl and -N(C1-4 alkyl)[(alkyl)-(heterocycle-substituted)-cycloalkyl];

R ₁₉ of formula (A-1) to formula (A-8) is selected from the group consisting of aryl, heteroaryl, bicyclic heteroaryl, and these aryl or heteroaryl groups may be substituted with halogen, C1-6 alkyl, C1-6 cycloalkyl, CF ₃ , F, CN, alkyne, alkyl sulfone, wherein the halogen substitution can be mono-bis or tri-substituted;

R ₂₀ and R ₂₁ of formula (A-1) - formula (A-8) are independently selected from C1-6 alkyl, C1-6 cycloalkyl, C1-6 alkoxy, hydroxylated C1-6 alkoxy, and fluoro-substituted C1-6 alkoxy, where R ₂₀ and R ₂₁ may further be linked to form a 5, 6 and 7 membered cyclic or heterocyclic ring, which may be further substituted;

R ₂₂ of formula (A-1) - formula (A-8) is selected from the group consisting of H, C1-6 alkyl, C1-6 cycloalkyl, carboxylic acid, carboxylic acid ester, amide, reverse amide, sulfonamide, reverse sulfonamide, N-acyl urea, a nitrogen-containing 5-membered heterocycle, wherein the 5-membered heterocycles can be additionally substituted with C1-6 alkyl, alkoxy, fluoro-alkyl, CN and alkylsulfone;

R ₂₃ of formula (A-1) - formula (A-8) is selected from aryl, heteroaryl, -O-aryl, -O-heteroaryl, -O-alkyl, -O-alkyl-cycloalkyl, -NH-alkyl, -NH -alkyl-cycloalkyl, -N(H)-aryl, -N(H)-heteroaryl, -N(alkyl)-aryl, -N(alkyl)-heteroaryl, wherein the aryl or heteroaryl groups may be substituted with halogen, C1- 6 alkyl, hydroxylated C1-6 alkyl, cycloalkyl, fluoro-substituted C1-6 alkyl, CN, alkoxy, alkyl sulfone, amide and sulfonamide;

R ₂₄ of formula (A-1) - formula (A-8) is selected from the group consisting of -CH ₂ -(C1-6 alkyl), -CH ₂ -cycloalkyl, -CH ₂ -aryl, CH ₂ -heteroaryl, where alkyl, cycloalkyl, aryl and heteroaryl may be substituted with halogen, alkoxy, hydroxylated alkyl, cyano-substituted alkyl, cycloalyl and substituted cycloalkyl;

R ₂₅ of formula (A-1) - formula (A-8) is selected from the group consisting of C1-6 alkyl, C1-6 alkyl-cycloalkyl, alkoxy-substituted alkyl, hydroxylated alkyl, aryl, heteroaryl, substituted aryl or heteroaryl, 5,6 and 7-membered nitrogen-containing saturated heterocycles, 5,6-fused and 6,6-fused nitrogen-containing saturated heterocycles and these saturated heterocycles can be substituted with C1-6 alkyl, fluoro-substituted C1-6 alkyl, alkoxy, aryl and heteroaryl group;

R ₂₆ of formula (A-1) - formula (A-8) is selected from the group consisting of C1-6 alkyl, C3-6 cycloalkyl, wherein the alkyl or cycloalkyl may be substituted by -OH, alkoxy, fluoro-substituted alkoxy, fluoro -substituted alkyl, -NH ₂ , -NH-alkyl, NH-C(O)alkyl, -NH-S(O) ₂ -alkyl and -S(O) ₂ -alkyl;

R ₂₇ of formula (A-1) - formula (A-8) is selected from the group consisting of aryl, heteroaryl, bicyclic heteroaryl, where the aryl or heteroaryl groups may be substituted with C1-6 alkyl, alkoxy, NH ₂ , NH-alkyl, halogen or -CN, and the substitution may be independently mono-, bis- or tri-substitution;

R ₂₈ of formula (A-1) - formula (A-8) is selected from the group consisting of aryl, 5 and 6 membered heteroaryl, bicyclic heteroaryl, cycloalkyl, saturated heterocycle such as piperidine, piperidinone, tetrahydropyran, N-acyl- piperidine, wherein cycloalkyl, saturated heterocycle, aryl or heteroaryl may be further substituted by -OH, alkoxy, mono-, bi- or tri-substituted, including halogen, -CN, alkyl sulfone and fluoro-substituted alkyl groups; And

R ₂₉ of formula (A-1) to formula (A-8) is selected from the group consisting of H, alkyl, aryl-substituted alkyl, alkoxy-substituted alkyl, cycloalkyl, aryl-substituted cycloalkyl and alkoxy-substituted cycloalkyl.

[00162] In certain embodiments, the heterocycles in R ^f and R ^g of formula (A-1) through formula (A-8) are substituted pyrrolidine, substituted piperidine, substituted piperisine.

[00163] More specifically, non-limiting examples of MLM include the molecules shown below, as well as those “hybrid” molecules that arise from a combination of 1 or more different properties shown in the molecules below.

[00164] Using the MLM in Formulas A-1 to A-8, the following PROTACs can be formulated to target a specific protein for degradation, where "L" represents a connector (i.e., a linker group) and "RTM" represents a ligand that binds to a target protein.

[00165] In certain embodiments, the disclosure provides a bifunctional molecule comprising a structure selected from the group consisting of:

where X, Ra, Y, Z, A, A', A'', R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ^b , R ^c , R ^d , R ₇ , R ^e , R ^f , R ^g , R ₉ , R ₁₀ , R ₁₁ , R ₁₂ , R ₁₃ , R ₁₄ , R ₁₅ , R ₁₆ , R ₁₇ , R ^k , R ^l , R ₁₈ , R ₁₉ , R ₂₀ , R ₂₁ , R ₂₂ , R ₂₃ , R ₂₄ , R ₂₅ , R ₂₆ , R ₂₇ , R ₂₈ and R _1'' are as defined herein with respect to formulas (A-1) to (A-8).

[00166] In certain embodiments, the disclosure suggests bifunctional or chimeric molecules with the structure: PTM-L-MLM, where PTM is a target protein binding moiety linked to the MLM by L, where L is a linkage (i.e., missing) or chemical linker. In certain embodiments, the MLM has a structure selected from the group consisting of A-1-1, A-1-2, A-1-3 and A-1-4:

Where:

R1' and R2' of formulas A-1-1 to A-1-4 (i.e., A-1-1, A-1-2, A-1-3, and A-1-4) are independently selected from the group consisting of F, Cl, Br, I, acetylene, CN, CF ₃ and NO ₂ ;

R3' is selected from the group consisting of -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH ₂ F, -OCH ₂ CH ₂ OCH ₃ and -OCH(CH ₃ ) ₂ ;

R4' of formulas A-1-1 - A-1-4 is selected from the group consisting of H, halogen, -CH ₃ , -CF ₃ , -OCH ₃ , -C(CH ₃ )3, -CH(CH ₃ ) ₂ , -cyclopropyl, -CN, -C(CH ₃ ) ₂ OH, -C(CH ₃ ) ₂ OCH ₂ CH ₃ , -C(CH ₃ ) ₂ CH ₂ OH, -C(CH ₃ ) ₂ CH ₂ OCH ₂ CH ₃ , C(CH ₃ ) ₂ CH ₂ OCH ₂ CH ₂ OH, -C(CH ₃ ) ₂ CH ₂ OCH ₂ CH ₃ , -C(CH ₃ ) ₂ CN, C(CH ₃ ) ₂ C(O )CH ₃ , -C(CH ₃ ) ₂ C(O) ₂ HCH ₃ , -C(CH ₃ ) ₂ C(O) ₂ (CH ₃ ) ₂ , -SCH ₃ , -SCH ₂ CH ₃ , -S( O) ₂ CH ₃ , -S(O ₂ )CH ₂ CH ₃ , -NHC(CH ₃ ) ₃ , -N(CH ₃ ) ₂ , pyrrolidinyl and 4-morpholinyl;

R5' of formulas A-1-1 - A-1-4 is selected from the group consisting of halogen, -cyclopropyl, -S(O) ₂ CH ₃ , -S(O) ₂ CH ₂ CH ₃ , 1-pyrrolidinyl, - NH ₂ , -N(CH ₃ ) ₂ and -NHC(CH ₃ ) ₃ ; And

R6' of formulas A-1-1 to A-1-4 is selected from the structures presented below, where the linker point is designated as "*".

In addition to R6' as a linker attachment point, R4' can also serve as a linker attachment position. In case R4' represents a linker junction, the linker will be connected to the terminal atom of the R4' groups shown above.

[00167] In certain embodiments, the attachment position of the linker of formulas A-1-1 to A-1-4 is at least one of R4' or R6', or both.

[00168] In certain embodiments, R6' of formulas A-1-1 through A-1-4 is independently selected from the group consisting of H,

Where denotes the linker attachment point.

[00169] In certain embodiments, a linker of formula A-4-1 to A-4-6 is attached to at least one of R1', R2', R3', R4', R5', R6', or a combination thereof.

[00170] In certain embodiments, the disclosure suggests bifunctional or chimeric molecules with the structure: PTM-L-MLM, where PTM is a target protein binding moiety linked to the MLM by L, where L is a linkage (i.e., missing) or chemical linker. In certain embodiments, the MLM has a structure selected from the group consisting of A-4-1, A-4-2, A-4-3, A-4-4, A-4.5 and A-4-6:

Where:

R7' of formula A-4-1 - A-4-6 (i.e., A-4-1, A-4-2, A-4-3, A-4-4, A-4-5, and A-4-6) is one or more (eg, 1, 2, 3, 4) halogens; R8' of formula A-4-1 - A-4-6 represents one or more groups (for example, 1, 2, 3 or 4 groups) selected from the group consisting of H, -F, -Cl, -Br, - I, -CN, -NO ₂ , ethylnyl, cyclopropyl, methyl, ethyl, isopropyl, vinyl, methoxy, ethoxy, isopropoxy, -OH, other C1-6 alkyl, other C1-6 alkenyl and C1-6 alkynyl, mono-, di- or trisubstituted; R9' of formula A-4-1 to A-4-6 is selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, alkenyl and substituted cycloalkenyl;

Z of formula A-4-1 - A-4-6 is selected from the group consisting of H, -OSH ₃ , -OSH ₂ CH ₃ and halogen;

R10' and R11' of formula A-4-1 to A-4-6 are each independently selected from the group consisting of H, (CH ₂ ) _n -R', (CH ₂ ) _n -NR'R'', (CH ₂ ) _n -NR'COR', (CH ₂ ) _n -NR'CO ₂ R'', (CH ₂ ) _n -COOH, (CH ₂ ) _n -COOR', (CH) _n -CONR'R'' , (CH ₂ ) _n -OR', (CH ₂ ) _n -SR', (CH ₂ ) _n -SOR', (CH ₂ ) _n -CH(OH)-R', (CH ₂ ) _n -COR' , (CH ₂ ) _n -SO ₂ R', (CH ₂ ) _n -SONR'R'', (CH ₂ ) _n -SO ₂ NR'R'', (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -R', (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -OH, (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -OR', (CH ₂ CH ₂ O) _m - (CH ₂ ) _n -NR'R'', (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -NR'COR'', (CH ₂ CH ₂ O) _m (CH ₂ ) _n -NR'SO ₂ R', (CH ₂ CH ₂ O) _m (CH ₂ ) _n COOH, (CH ₂ CH ₂ O) _m (CH ₂ ) _n -COOR', (CH ₂ CH ₂ O) _m -(CH ₂ ) ₂ -CONR'R'', (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -SO ₂ R', (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -COR', (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -SONR'R'', (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -SO ₂ NR'R'', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n R', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -OH, (CH ₂ ) _p -(CH ₂ CH ₂ O) _m - (CH ₂ ) _n -OR', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m (CH ₂ ) _n , (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n - NR'COR'', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -NR'SO ₂ R'', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m - (CH ₂ ) _n -COOH, (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -COOR', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -CONRR'', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -SO ₂ R', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -COR', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -SONR'R'', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -( CH ₂ ) _n -SO ₂ NR'R _m , aryl-(CH ₂ ) _n -COOH, and heteroaryl-alkyl-CO-alkyl-NR'R'm, where alkyl may be substituted with OR, and heteroaryl-(CH ₂ ) _n -heterocycle, where the heterocycle may optionally be substituted with alkyl, hydroxyl, COOR and COR; where R' and R'' are selected from H, alkyl, halogen-substituted alkyl, hydroxyl, NH ₂ , NH(alkyl), N(alkyl) ₂ , oxo, carboxy, cycloalkyl and heteroaryl; m, n and p independently represent 0-6;

R12' of formula A-4-1 to A-4-6 is selected from the group consisting of -O-(alkyl), -O-(alkyl)-alkoxy, -C(O)-(alkyl), -C(OH )-alkyl-alkoxy, -C(O)-NH-(alkyl), -C(O)-N-(alkyl) ₂ , -S(O)-(alkyl), S(O) ₂ -(alkyl) , -C(O)-(cyclic amine) and -O-aryl-(alkyl), -O-aryl-(alkoxy);

R1'' of formula A-4-1 to A-4-6 is selected from the group consisting of H, alkyl, aryl-substituted alkyl, alkoxy-substituted alkyl, cycloalkyl, aryl-substituted cycloalkyl and alkoxy-substituted cycloalkyl.

[00171] In any of the aspects or embodiments described herein, alkyl, alkoxy, or the like. may be lower alkyl or lower alkoxy.

[00172] In certain embodiments, the attachment position of the linker of formula A-4-1-A-4-6 is at least one of Z, R8', R9', R10', R11'', R12'', or R1' '.

[00173] The method used for the design of chimeric molecules as presented in A-1-1-A-1-4, A-4-1-A-4-6 can be applied to MLM with formula A-2, A-3 , A-5, A-6, A-7 and A-8, where the solvent exposed region of the MLM can be linked to a linker "L" which will be attached to the target protein ligand "PTM", for the PROTAC construct.

[00174] In any aspect or embodiment described herein, the MLM is selected from:

[00175] Typical MDM2 binding fragments include, but are not limited to, the following:

[00176] 1. HDM2/MDM2 inhibitors defined in Vassilev, et ai, In vivo activation of the p53 pathway by small-molecule antagonists of MDM2, SCIENCE vol:303, pag:844-848 (2004) and Schneekloth, et al , Targeted intracellular protein degradation induced by a small molecule: En route to chemical proteomics, Bioorg. Med. Chem. Lett. 18 (2008) 5904-5908, including (or additionally) the compounds nutlin-3, nutlin-2 and nutlin-1 (derivatives) as described below, as well as all derivatives and analogs thereof:

(derivatives wherein the linker group L or the -(L-MLM) group is attached, for example, to a methoxy group or as a hydroxyl group);

(derivatives wherein the linker group L or the -(L-MLM) group is attached to, for example, a methoxy group or a hydroxyl group);

(derivatives wherein the linker group L or the -(L-MLM) group is attached, for example through a methoxy group or as a hydroxyl group); And

[00177] 2. Trans-4-iodo-4'-boranyl-chalcone

[00178]

[00179] (derivatives wherein the linker group L or the linker group L or the -(L-MLM) group is attached, for example, through a hydroxyl group).

[00180] Typical CLMs

[00181] Neoimide compounds

[00182] In one aspect, the disclosure provides compounds useful for binding and/or inhibiting cereblon. In certain embodiments, the compound is selected from the group consisting of chemical structures:

Where:

W of formulas (a) - (f) is independently selected from the group of CH ₂ , CHR, C=O, SO ₂ , NH and N-alkyl;

X of formulas (a) - (f) is independently selected from the group O, S and H ₂ ;

Y of formulas (a) - (f) is independently selected from the group CH ₂ , -C=CR', NH, N-alkyl,

N-aryl, N-hetaryl, N-cycloalkyl, N-heterocyclyl, O and S;

Z of formulas (a) - (f) is independently selected from the group O and S or H ₂ except

that both X and Z cannot be H ₂ ;

G and G' of formulas (a) - (f) are independently selected from the group H, optionally substituted linear or branched alkyl, OH, R'OCOOR, R'OCONRR'', CH ₂ -heterocyclyl, optionally substituted R', and benzyl, optionally substituted R';

Q1 - Q4 of formulas (a) - (f) represent a C carbon substituted by a group independently selected from R', N or N-oxide;

A of formulas (a) - (f) is independently selected from the group H, optionally substituted linear or branched alkyl, cycloalkyl, Cl and F;

R of formulas (a) - (f) includes, but is not limited to: -CONR'R'', -OR', -NR'R'', -SR', -SO ₂ R', -SO ₂ NR'R '', -CR'R''-, -CR'NR'R''-, (-CR'O) _n R'', -aryl, -hetaryl, -optionally substituted linear or branched alkyl, -cycloalkyl, - heterocyclyl, -P(O)(OR')R'', -P(O)R'R'', -OP(O)(OR')R'', -OP(O)R'R'', -Cl, -F, -Br, -I, -CF ₃ , -CN, -NR'SO ₂ NR'R'', -NR'CONR'R'', -CONR'COR'', -NR,C (=N-CN) ₂ R,R'', -C(=N-CN) ₂ R'R'', -NR'C(=N-CN)R'', -NR'C(=C- NO ₂ ) ₂ R'R'', -SO ₂ NR'COR'', -NO ₂ , -CO ₂ R', -C(C=N-OR')R'', -CR'=CR'R '', -CCR', -S(C=O)(C=N-R')R'', -SF5 and -OCF ₃

R' and R'' of formulas (a) - (f) are independently selected from H, alkyl, cycloalkyl, aryl, heteroaryl, heterocycle, -C(=O)R, heterocyclyl, each of which is optionally substituted;

R _n of formulas (a) - (f) is an integer from 1-10 (eg 1-4, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10);

formulas (a) - (f) represent a bond that may be stereospecific ((R) or (S)) or non-stereospecific; and Rn of formulas (a) - (f) contains 1-4 independently selected functional groups or atoms, for example, O, OH, N, C1-C6 alkyl, C1-C6 alkoxy, -alkyl-aryl (for example, -alkyl-aryl containing at least one of C1-C6 alkyl, C5-C7 aryl or combinations thereof), aryl (for example, C5-C7 aryl), amine, amide or carboxy.

[00183] Typical CLMs

[00184] In any of the compounds described herein, the CLM contains a chemical structure selected from the group:

Where:

W of formulas (a) - (f) is independently selected from the group of CH ₂ , CHR, C=O, SO ₂ , NH and N-alkyl;

X of formulas (a) - (f) is independently selected from the group O, S and H ₂ ;

Y of formulas (a) - (f) is independently selected from the group CH ₂ , -C=CR', NH, N-alkyl,

N-aryl, N-hetaryl, N-cycloalkyl, N-heterocyclyl, O and S;

Z of formulas (a) - (f) is independently selected from the group O and S or H ₂ except

that both X and Z cannot be H ₂ ;

G and G' of formulas (a) - (f) are independently selected from the group H, optionally substituted linear or branched alkyl, OH, R'OCOOR, R'OCONRR'', CH ₂ -heterocyclyl, optionally substituted R', and benzyl, optionally substituted R';

C1 - C4 of formulas (a) - (f) represent a C carbon substituted by a group independently selected from R', N or N-oxide;

A of formulas (a) - (f) is independently selected from the group H, alkyl (linear, branched, optionally substituted), cycloalkyl, C1 and F;

R of formulas (a) - (f) includes, but is not limited to: -CONR'R'', -OR', -NR'R'', -SR', -SO ₂ R', -SO ₂ NR'R '', -CR'R''-, -CR'NR'R''-, (-CR'OVR'', -aryl, -hetaryl, -alkyl, -cycloalkyl, -heterocyclyl, -P(O)( OR')R'', -P(O)R'R'', -OP(O)(OR')R'', -OP(O)R'R'', -Cl, -F, -Br , -I, -CF ₃ , -CN, -NR,SO ₂ NR'R'', -NR'CONR'R'', -CONR'COR'', -NR'C(=N-CN) ₂ R 'R'', -C(=N-CN) ₂ R'R'', -NR'C(=N-CN)R'', -NR,C(=C-NO ₂ ) ₂ R,R'', -SO ₂ NR'COR'', -NO ₂ , -CO ₂ R', -C(C=N-OR')R'', -CR'=CR'R'', -CCR', - S(C=O)(C=N-R')R'', -SF5 and -OCF ₃ R' and R'' of formulas (a) - (f) are independently selected from bond, H, alkyl, cycloalkyl, aryl , heteroaryl, heterocycle, -C(=O)R, heterocyclyl, each of which is optionally substituted;

n of formulas (a) - (f) is an integer from 1-10 (for example, 1-4, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10);

formulas (a) - (f) represent a bond that may be stereospecific ((R) or (S)) or non-stereospecific; and Rn of formulas (a) - (f) contains 1-4 independently selected functional groups or atoms, for example, O, OH, N, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, -alkyl-aryl (for example, -alkyl-aryl containing at least one of C ₁ -C ₆ alkyl, C ₅ -C ₇ aryl or combinations thereof), aryl (for example, C ₅ -C ₇ aryl), amine, amide or carboxy, and optionally, one of which is modified to be covalently linked to a PTM, a chemical linker group (L), ULM, CLM (or CLM'), or a combination thereof.

[00185] In certain embodiments described herein, the CLM or ULM contains a chemical structure selected from the group:

formula (g)

Where:

W of formula (g) is independently selected from the group CH ₂ , C=O, NH and N-alkyl; R of formula (g) is independently selected from a H, methyl, or optionally substituted straight or branched alkyl (eg, optionally substituted straight or branched C1-C6 alkyl);

formula (g) represents a bond that may be stereospecific ((R) or (S)) or non-stereospecific; And

Rn of formula (g) contains 1 to 4 independently selected functional groups or atoms, for example, O, OH, N, C1-C6 alkyl, C1-C6 alkoxy, -alkyl-aryl (for example, -alkyl-aryl containing at least one of C1-C6 alkyl, C5-C7 aryl, or combinations thereof), aryl (e.g., C5-C7 aryl), amine, amide, or carboxy, and optionally one of which is modified to be covalently linked to a PTM, chemical linker group (L), ULM, CLM (or CLM'), or a combination thereof.

[00186] In any of the embodiments described herein, W, X, Y, Z, G, G', R, R, R'', C1-Q4, A and Rn of formulas (a) - (g) may be independently covalently linked to a linker and/or a linker to which one or more PTM, ULM, CLM or CLM1 groups are attached.

[00187] In any of the aspects or embodiments described herein, Rn contains 1-4 independently selected functional groups or atoms, for example, O, OH, N, C1-C6 alkyl, C1-C6 alkoxy, -alkyl-aryl (e.g., -alkyl-aryl containing at least one of C1-C6 alkyl, C5-C7 aryl, or combinations thereof), aryl (e.g., C5-C7 aryl), amine, amide, or carboxy, and optionally one of which modified to be covalently linked to a PTM, a chemical linker group (L), ULM, CLM (or CLM'), or a combination thereof.

[00188] In any of the aspects or embodiments described herein, Rn contains 1-4 functional groups and atoms, for example, O, OH, N, C1-C6 alkyl, C1-C6 alkoxy, amine, amide, or carboxy , and optionally, one of which is modified to be covalently linked to a PTM, a chemical linker (L) group, a ULM, a CLM (or CLM'), or a combination thereof.

[00189] More specifically, non-limiting examples of CLMs include the molecules shown below, as well as those “hybrid” molecules that arise from a combination of 1 or more different properties shown in the molecules below.

[00190] In any of the compounds described herein, the CLM contains a chemical structure selected from the group:

Where:

W is independently selected from CH ₂ , CHR, C=O, SO ₂ , NH and N-alkyl;

Q ₁ , Q ₂ , Q ₃ , Q ₄ , Q ₅ each independently represent a C or N carbon substituted with a group independently selected from R', N or N-oxide;

R ¹ is selected from: none, H, OH, CN, C1-C3 alkyl, C=O;

R ² is selected from the group: absence, H, OH, CN, C1-C3 alkyl, CHF ₂ , CF ₃ , CHO, C(=O)NH ₂ ;

R ³ is selected from H, alkyl (for example, C1-C6 or C1-C3 alkyl), substituted alkyl (for example, substituted C1-C6 or C1-C3 alkyl), alkoxy (for example, C1-C6 or C1-C3 alkoxy), substituted alkoxy (eg substituted C1-C6 or C1-C3 alkoxy);

R ⁴ is selected from H, alkyl, substituted alkyl;

R ⁵ and R ⁶ are each independently H, halogen, C(=O)R', CN, OH, CF ₃ ;

X represents C, CH, C=O or N;

X ₁ represents C=O, N, CH or CH ₂ ;

R' is selected from H, halogen, amine, alkyl (eg C1-C3 alkyl), substituted alkyl (eg substituted C1-C3 alkyl), alkoxy (eg C1-C3 alkoxyl), substituted alkoxy (eg substituted C1- C3 alkoxyl), NR ² R ³ , C(=O)OR ² , optionally substituted phenyl;

n represents 0-4;

represents a single or double bond; And

CLM is covalently linked to PTM, a chemical linker group (L), ULM, CLM (or CLM'), or a combination thereof.

[00191] In any aspect or embodiment described herein, CLM or CLM' is covalently linked to a PTM, a chemical linker (L) group, ULM, CLM, CLM', or a combination thereof via an R group (such as R, R ¹ , R ² , R ³ , R ⁴ or R'), W, X, or a Q group (such as Q ₁ , Q ₂ , Q ₃ , Q ₄ or Q ₅ ).

[00192] In any of the embodiments described herein, CLM or CLM' is covalently linked to a PTM, a chemical linker group (L), ULM, CLM, CLM', or a combination thereof via W, X, R, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R', Q ₁ , Q ₂ , Q ₃ , Q ₄ and Q ₅ .

[00193] In any of the embodiments described herein, W, X, R ¹ , R ² , R ³ , R ⁴ , R', Q ₁ , Q ₂ , Q ₃ , Q ₄ , and Q ₅ may be independently covalently linked to a linker and/or a linker to which one or more PTM, ULM, ULM', CLM or CLM' groups are attached.

[00194] More specifically, non-limiting examples of CLMs include those shown below, as well as those “hybrid” molecules or compounds that arise from the combination of 1 or more different properties of the following compounds:

Where:

W is independently selected from the group CH ₂ , CHR, C=O, SO ₂ , NH and N-alkyl;

R ¹ is selected from the group: absence, H, CH, CN, C1-C3 alkyl;

R ² represents H or C1-C3 alkyl;

R ³ is selected from H, alkyl, substituted alkyl, alkoxy, substituted alkoxy;

R ⁴ is methyl or ethyl;

R ⁵ represents H or halogen;

R ⁶ represents H or halogen;

R in CLM is H;

R' represents H or the point of attachment for PTM, PTM', chemical linker group (L), ULM, CLM, CLM',

Q ₁ and Q ₂ are each independently C or N substituted with a group independently selected from H or C1-C3 alkyl;

represents a single or double bond; And

Rn contains a functional group or atom.

[00195] In any of the embodiments described herein, W, R ¹ , R ² , Q ₁ , Q ₂ , Q ₃ , Q _{4 ,} and Rn may be independently covalently linked to the linker and/or the linker to which the one or more groups of PTM, ULM, ULM', CLM or CLM'.

[00196] In any of the embodiments described herein, R ¹ , R ² , Q ₁ , Q ₂ , Q ₃ , Q _{4 ,} and Rn may be independently covalently linked to a linker and/or a linker to which one or more more than RTM, ULM, ULM', CLM or CLM' groups.

[00197] In any of the embodiments described herein, Q ₁ , Q ₂ , Q ₃ , Q ₄ and Rn can be independently covalently linked to a linker and/or a linker to which is attached one or more PTM, ULM, ULM', CLM or CLM'.

[00198] In any aspect or embodiment described herein, R _n is modified to be covalently linked to a linker group (L), PTM, ULM, a second CLM having the same chemical structure as the CLM, CLM', a second linker, or any several of them, or a combination thereof.

[00199] In any aspect or embodiment described herein, the CLM is selected from:

wherein R' is halogen and R ¹ is as described in any aspect or embodiment described herein.

[00200] In some cases, "CLM" may be imides that bind to the E3 ligase cereblon. These imides and linker attachment points can be, but are not limited to, the following structures:

[00201] Typical VLMs

[00202] In certain embodiments of the compounds as described herein, the ULM is a VLM and contains a chemical structure selected from the group ULM-a:

,

Where:

the dotted line denotes the attachment of at least one PTM, another ULM, or VLM, or MLM, or ILM, or CLM (i.e., ULM', or VLM', or CLM', or ILM', or MLM'), or a fragment of a chemical linker linking at least one PTM, ULM', or VLM', or CLM', or ILM' or MLM' to the other end of the linker;

X ¹ , X ² of the ULM-a formula are each independently selected from the bond group, O, NR ^Y3 , CR ^Y3 R ^Y4 , C=O, C=S, SO and SO ₂ ;

R ^Y3 , R ^Y4 of formula ULM-a are each independently selected from the group H, linear or branched C _1-6 alkyl, optionally substituted with 1 or more halogens, optionally substituted C _1-6 alkoxy (e.g., optionally substituted with 0- 3 R ^P groups);

R ^P of formula ULM-a represents 0, 1, 2 or 3 groups, each independently selected from the group H, halogen, -OH, C _1-3 alkyl, C=O;

W ³ of formula ULM-a is selected from the group of optionally substituted T, optionally substituted -TN(R ^1a R ^1b )X ³ , optionally substituted -T-N(R ^1a R ^1b ), optionally substituted -T-aryl, optionally substituted -T -heteroaryl, optionally substituted T-biheteroaryl, optionally substituted -T-heterocyclyl, optionally substituted -T-biheterocyclyl, optionally substituted -NR ¹ -T-aryl, optionally substituted -NR ¹ -T-heteroaryl or optionally substituted -NR ¹ -T -heterocycle;

X ³ of the formula ULM-a represents C=O, R ¹ , R ^1a , R ^1b ;

each of R ¹ , R ^1a , R ^1b is independently selected from the group consisting of an H, straight or branched C ₁ -C ₆ alkyl group, optionally substituted with 1 or more halogens or -OH groups, R ^Y3 C=O, R ^Y3 C=S, R ^Y3 SO, R ^Y3 SO ₂ , N(R ^Y3 R ^Y4 )C=O, N(R ^Y3 R ^Y4 )C=S, N(R ^Y3 R ^Y4 )SO and N(R ^Y3 R ^Y4 )SO ₂ ;

T of formula ULM-a is selected from the group of optionally substituted alkyl, - (CH ₂ ) _n - group, where each of the methylene groups is substituted by one or two substituents selected from the group of halogen, methyl, linear or branched C ₁ -C ₆ alkyl group, optionally substituted 1 or more halogens, C(O)NR ¹ R ^1a , or NR ¹ R ^1a , or R ¹ and R ^1a are associated with the formation of an optionally substituted heterocyclyl, or -OH groups, or an optionally substituted amino acid side chain;

W ⁴ of formula ULM-a is an optionally substituted -NR1-T-aryl group, where the aryl group may be an optionally substituted 5-6 membered heteroaryl group, an optionally substituted -NR1-T-heteroaryl group, or an optionally substituted -NR1-T-heterocycle where -NR1 is covalently linked to X ² and R ¹ represents H or CH ₃ , preferably H; And

n is 0-6, often 0, 1, 2 or 3, preferably 0 or 1.

[00203] In any of the embodiments described herein, T is selected from the group of optionally substituted alkyl, -(CH ₂ ) _n- group, wherein each of the methylene groups is optionally substituted by one or two substituents selected from the group of halogen, methyl, optionally substituted alkoxy, linear or branched C ₁ -C ₆ alkyl group, optionally substituted with 1 or more halogens, C(O)NR ¹ R ^1a , or NR ¹ R ^1a , or R ¹ and R ^1a are associated with the formation of an optionally substituted heterocycle, or -OH groups, or an optionally substituted amino acid side chain; and n is 0-6, often 0, 1, 2 or 3, preferably 0 or 1.

[00204] In certain embodiments, W ⁴ of formula ULM-a is where W ⁵ is optionally substituted phenyl or optionally substituted 5-10 membered heteroaryl (e.g., optionally substituted with one or more [such as 1, 2, 3, 4 or 5] halogens, CN, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkoxy, hydroxy or optionally substituted haloalkoxy), and R _14a , R _14b , each independently selected from the group H, haloalkyl or optionally substituted alkyl.

[00205] In any of the embodiments, W ⁵ of formula ULM-a is selected from the group of optionally substituted phenyl or optionally substituted 5-10 membered heteroaryl (e.g., W ⁵ is optionally substituted with one or more [such as 1, 2, 3, 4 or 5] halogens, CN, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkoxy, hydroxy or optionally substituted haloalkoxy), and

R ₁₅ of formula ULM-a is selected from the group H, halogen, CN, OH, NO ₂ , N R1 _4a R _14b , OR _14a , CONR _14a R _14b , NR _14a COR _14b , SO ₂ NR _14a R _14b , NR _14a SO ₂ R _14b , optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl or optionally substituted cycloheteroalkyl;

[00206] In additional embodiments, W ⁴ substituents for use herein also specifically include (and without limitation to the specific compound disclosed) W ⁴ substituents that are found in the identified compounds disclosed herein. Each of these W ⁴ substituents can be used in combination with any number of W ³ substituents, which are also disclosed herein.

[00207] In certain additional embodiments, ULM-a is optionally substituted with 0-3 R ^P groups on the pyrrolidine moiety, each R ^P independently representing H, halogen, -OH, C1-3 alkyl, C=O.

[00208] In any of the embodiments described herein, W ³ , W ⁴ of formula ULM-a may be independently covalently linked to a linker that attaches one or more PTM groups, and wherein the dotted line represents the site of attachment of at least one A PTM, another ULM (ULM'), or a chemical linker moiety linked to at least one PTM or ULM', or both ULMs.

[00209] In certain embodiments, the ULM is a VHL and is represented by the structure:

Where:

W ³ of formula ULM-b is selected from the group of optionally substituted aryl, optionally substituted heteroaryl, or

R ₉ and R ₁₀ of formula ULM-b are independently hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted hydroxyalkyl, optionally substituted heteroaryl or haloalkyl, or R ₉ , R ₁₀ and the carbon atom to which they are attached form an optionally substituted cycloalkyl;

R ₁₁ of formula ULM-b is selected from the group of optionally substituted
heterocycle, optionally substituted alkoxy, optionally substituted heteroaryl, optionally substituted aryl, ,

R ₁₂ of formula ULM-b is selected from the group H or optionally substituted alkyl;

R ₁₃ of formula ULM-b is selected from the group H, optionally substituted alkyl, optionally substituted alkylcarbonyl, optionally substituted (cycloalkyl)alkylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted arylcarbonyl, optionally substituted (heterocyclyl)carbonyl, or optionally substituted aralkyl;

R _14a , R _14b of formula ULM-b, each independently selected from the group H, haloalkyl or optionally substituted alkyl;

W ⁵ of formula ULM-b is selected from the group of optionally substituted phenyl or optionally substituted 5-10 membered heteroaryl (e.g., W ⁵ is optionally substituted with one or more [such as 1, 2, 3, 4 or 5] halogens, CN, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkoxy, hydroxy or optionally substituted haloalkoxy),

R ₁₅ of formula ULM-b is selected from the group H, halogen, CN, OH, NO ₂ , NR _14a R _14b , OR _14a , CONR _14a R _14b , NR _14a COR _14b , SO ₂ NR _14a R _14b , NR _14a SO ₂ R _14b , optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy; aryl, heteroaryl, cycloalkyl or cycloheteroalkyl (each optionally substituted);

each R ₁₆ of formula ULM-b is independently selected from the group halogen, CN, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkoxy, hydroxy or optionally substituted haloalkoxy;

o of the formula ULM-b is 0,1,2,3 or 4;

R ₁₈ of formula ULM-b is independently selected from the group H, halogen, optionally substituted alkoxy, cyano, optionally substituted alkyl, haloalkyl, haloalkoxy or a linker; And

p of formula ULM-b is 0, 1, 2, 3, or 4, and wherein the dotted line represents the attachment site of at least one PLM, another ULM (ULM'), or a chemical linker moiety associated with at least one PTM or ULM ', or with both ULMs.

[00210] In certain embodiments, R ₁₅ of formula ULM-b is where R ₁₇ represents H, halogen, optionally substituted C _3-6 cycloalkyl, optionally substituted C _1-6 alkyl, optionally substituted C _1-6 alkenyl and C _1-6 haloalkyl; and Ha represents S or O.

[00211] In certain embodiments, R ₁₇ of formula ULM-b is selected from the group of methyl, ethyl, isopropyl, and cyclopropyl.

[00212] In certain additional embodiments, R ₁₅ of formula ULM-b is selected from the group consisting of:

[00213] In certain embodiments, R ₁₁ of formula ULM-b is selected from the group consisting of:

[00214] In certain embodiments, the ULM has a chemical structure selected from the group:

Where:

R ₁ of formulas ULM-c, ULM-d and ULM-e is H, ethyl, isopropyl, t-butyl, sec-butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; optionally substituted alkyl, optionally substituted hydroxyalkyl, optionally substituted heteroaryl or haloalkyl;

R _14a of formulas ULM-c, ULM-d and ULM-e is H, haloalkyl, optionally substituted alkyl, methyl, fluoromethyl, hydroxymethyl, ethyl, isopropyl or cyclopropyl;

R ₁₅ of formulas ULM-c, ULM-d and ULM-e is selected from the group consisting of H, halogen, CN, OH, NO ₂ , optionally substituted heteroaryl, optionally substituted aryl; optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy, optionally substituted cycloalkyl or optionally substituted cycloheteroalkyl;

X of formulas ULM-c, ULM-d and ULM-e is C, CH ₂ or C=O

R ₃ of formulas ULM-c, ULM-d and ULM-e is absent or is an optionally substituted 5 or 6 membered heteroaryl; And

the dotted line represents the attachment site of at least one PTM, another ULM (ULM'), or a chemical linker moiety associated with at least one PTM or ULM', or both ULMs.

[00215] In certain embodiments, the ULM contains a group in accordance with the chemical structure:

Where:

R _14a of formula ULM-f is H, haloalkyl, optionally substituted alkyl, methyl, fluoromethyl, hydroxymethyl, ethyl, isopropyl or cyclopropyl;

R ₉ of formula ULM-f is H;

R ₁₀ of formula ULM-f is H, ethyl, isopropyl, tert-butyl, sec-butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl;

R ₁₁ of the formula ULM-f is

or optionally substituted heteroaryl;

p of formula ULM-f is 0, 1, 2, 3 or 4;

each R ₁₈ of formula ULM-f independently represents halogen, optionally substituted alkoxy, cyano, optionally substituted alkyl, haloalkyl, haloalkoxy or a linker;

R ₁₂ of the formula ULM-f represents H, C=O;

R ₁₃ of formula ULM-f is H, optionally substituted alkyl, optionally substituted alkylcarbonyl, optionally substituted (cycloalkyl)alkylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted arylcarbonyl, optionally substituted (heterocyclyl)carbonyl or optionally substituted aralkyl,

R ₁₅ of formula ULM-f is selected from the group consisting of H, halogen, Cl, CN, OH, NO ₂ , optionally substituted haloalkyl, optionally substituted heteroaryl, optionally substituted aryl;

wherein the dotted line of formula ULM-f denotes the site of attachment of at least one PTM, another ULM (ULM'), or a chemical linker moiety associated with at least one PTM or ULM', or both ULMs.

[00216] In certain embodiments, the ULM is selected from the following structures:

where n represents 0 or 1.

[00217] In certain embodiments, the ULM is selected from the following structures:

where the phenyl ring in ULM-a1 - ULM-a15, ULM-b1 - ULM-b12, ULM-c1 - ULM-c 15 and ULM-d1 - ULM-d9 is optionally substituted with fluorine, lower alkyl and alkoxy groups, and where the dotted line denotes the attachment site of at least one PTM, another ULM (ULM5), or a chemical linker moiety associated with at least one PTM, or ULM5, or both ULM-a.

[00218] In one embodiment, the phenyl ring in ULM-a1 - ULM-a15, ULM-b1 - ULM-b12, ULM-c1 - ULM-c15 and ULM-d1 - ULM-d9 can be functionalized as an ester to make part of it is about medications.

[00219] In certain embodiments, the hydroxyl group on the pyrrolidine ring of ULM-a1 - ULM-a15, ULM-b1 - ULM-b12, ULM-c1 - ULM-c15 and ULM-d1 - ULM-d9, respectively, contains a prodrug moiety, bound to an ester.

[00220] In any of the aspects or embodiments described herein, ULM and, where present, ULM' are each independently a group according to chemical structure:

or a pharmaceutically acceptable salt thereof, where:

R ^1' in ULM-g represents an optionally substituted C ₁ -C ₆ alkyl group, optionally substituted -(CH ₂ ) _n OH, optionally substituted - (CH ₂ ) _n SH, optionally substituted (CH ₂ ) _n -O-( a C ₁ -C ₆ )alkyl group, an optionally substituted (CH ₂ ) _n -WCOCW-(C ₀ -C ₆ )alkyl group containing a WCOCW epoxy moiety, where each W independently represents an H or a C ₁ -C ₃ alkyl group, optionally substituted -(CH ₂ ) _n COOH, optionally substituted -(CH ₂ ) _n C(O)-(C ₁ -C ₆ alkyl), optionally substituted -(CH ₂ ) _n NHC(O)-R ₁ , optionally substituted -(CH ₂ ) _n C(O)-NR ₁ R ₂ , optionally substituted -(CH ₂ ) _n OC(O)- NR ₁ R ₂ , -(CH ₂ O) _n H, optionally substituted -(CH ₂ ) _n OC(O)-(C ₁ -C ₆ alkyl), optionally substituted -(CH ₂ ) _n C(O)-O-(C ₁ -C ₆ alkyl), optionally substituted -(CH ₂ O) _n COOH, optionally substituted -(OCH ₂ ) _n O-(C ₁ -C ₆ alkyl), optionally substituted -(CH ₂ O) _n C(O)-(C ₁ -C ₆ alkyl), optionally substituted -(OCH ₂ ) _n NHC(O)-R ₁ , optionally substituted -(CH ₂ O) _n C(O)-NR ₁ R ₂ , -(CH ₂ CH ₂ O) _n H, optionally substituted (CH ₂ CH ₂ O) _n COOH, optionally substituted -(OCH ₂ CH ₂ ) _n O-(C ₁ -C ₆ alkyl), optionally substituted -(CH ₂ CH ₂ O) _n C(O)-(C ₁ -C ₆ alkyl), optionally substituted -( OCH ₂ CH ₂ ) _n NHC(O)-R ₁ , optionally substituted (CH ₂ CH ₂ O) _n C(O)-NR ₁ R ₂ , optionally substituted -SO ₂ R _S , optionally substituted S(O)R _S , NO ₂ , CN or halogen (F, Cl, Br, I, preferably F or Cl);

R ₁ and R ₂ in ULM-g each independently represent H or a C ₁ -C ₆ alkyl group, which may be optionally substituted by one or two hydroxyl groups or up to three halogen groups (preferably fluorine);

R _s in ULM-g represents a C ₁ -C ₆ alkyl group, an optionally substituted aryl, heteroaryl, or heterocyclic group, or a -(CH ₂ ) _m NR ₁ R ₂ group;

X and X' in ULM-g are each independently C=O, C=S, -S(O), S(O) ₂ (preferably X and X' are both C=O);

R ^2' in ULM-g represents an optionally substituted -(CH ₂ ) _n - (C=O) _u (NR ₁ ) _v (SO ₂ ) _w alkyl group, optionally substituted -(CH ₂ ) _n -(C=O ) _u (NR ₁ ) _v (SO ₂ ) _w NR _1N R _2N group, optionally substituted -(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -aryl, optionally substituted -(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -heteroaryl, optionally substituted -(CH ₂ ) _n -(C=O) _v NR ₁ (SO ₂ ) _w -heterocycle , optionally substituted -NR ₁ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -alkyl, optionally substituted -NR ¹ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -NR _1N R _2N , optionally substituted -NR ¹ -(CH ₂ ) _n - C(O) _u (NR ₁ ) _v (SO ₂ ) _w -NR ₁ C(O)R ₁ N, optionally substituted -NR ¹ -(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -aryl, optionally substituted -NR ¹ -(CH ₂ ) _n -aryl-heteroaryl, optional substituted -NR ¹ -(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -heteroaryl or optionally substituted -NR ¹ -(CH ₂ ) _n -(C=O) _v NR ₁ (SO ₂ ) _w -heterocycle, optionally substituted -X ^R2' -alkyl group; an optionally substituted -X ^R2' - aryl group; an optionally substituted -X ^R2' - heteroaryl group; an optionally substituted -X ^R2' heteropyclic group; optionally substituted;

R ^3' in ULM-g is optionally substituted alkyl, optionally substituted -(CH ₂ ) _n -(O) _u (NR ₁ ) _v (SO ₂ ) _w -alkyl, optionally substituted -(CH ₂ ) _n -C( O) _u (NR ₁ ) _v (SO ₂ ) _w -NR _1N R _2N , optionally substituted -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -NR ₁ C(O) R _1N , optionally substituted -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -C(O)NR ₁ R ₂ , optionally substituted -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -apyl, optionally substituted -(CH ₂ ) _n - C(O) _u (NR ₁ ) _v (SO ₂ ) _w -heteroaryl, optionally substituted -(CH ₂ ) _n - C(O) _u (NR ₁ ) _v (SO ₂ ) _w -heterocycle, optionally substituted -NR ¹ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -alkyl, optionally substituted -NR ¹ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -NR _1N R _2N , optionally substituted -NR ¹ -(CH ₂ ) _n - C(O) _u (NR ₁ ) _v (SO ₂ ) _w -NR ₁ C(O)R _1N , optionally substituted -NR ¹ -(CH ₂ ) _n - C(O) _u (NR ₁ ) _v (SO ₂ ) _w -aryl, optionally substituted -NR ¹ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -heteroaryl, optionally substituted -NR ¹ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -heterocycle, optionally substituted -O-(CH ₂ ) _n - (C=O) _u (NR ₁ ) _v (SO ₂ ) _w -alkyl, optionally substituted -O-(CH ₂ ) _n -( C=O) _u (NR ₁ ) _v (SO ₂ ) _w -NR _1N R _2N , optionally substituted -O-(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -NR ₁ C(O)R _1N , optionally substituted -O-(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -aryl, optionally substituted -O-(CH ₂ ) _n - ( C=O) _u (NR ₁ ) _v (SO ₂ ) _w -heteroaryl or optionally substituted -O-(CH ₂ ) _n - (C=O) _u (NR ₁ ) _v (SO ₂ ) _w -heterocycle; -(CH ₂ ) _n -(V) _n' -(CH ₂ ) _n -(V) _n' -alkyl group, optionally substituted -(CH ₂ ) _n -(V) _n' -(CH ₂ ) _n -( V) _n' -aryl group, optionally substituted -(CH ₂ ) _n -(V) _n' -(CH ₂ ) _n -(V) _n' -heteroaryl group, optionally substituted -(CH ₂ ) _n -(V) _n' -(CH ₂ ) _n -(V) _n' - heterocyclic group, optionally substituted -(CH ₂ ) _n -N(R _1' )(C=O) _m' -(V) _n' -alkyl group, optionally substituted -(CH ₂ ) _n -N(R _1' )(C=O) _m' -(V)n'-aryl group, optionally substituted -(CH ₂ ) _n -N(R _1' )(C= O) _m' -(V) _n' -heteroaryl group, optionally substituted - (CH ₂ ) _n -N(R _1' )(C=O) _m' -(V) _n' -heterocyclic group, optionally substituted -X ^R3' - alkyl group; an optionally substituted -X ^R3' - aryl group; an optionally substituted -X ^R3' - heteroaryl group; optionally substituted -X ^R3' - heterocyclic group;

R _1N and R _2N in ULM-g are each independently H, C ₁ -C ₆ alkyl, which is optionally substituted with one or two hydroxyl groups and up to three halogen groups, or optionally substituted -(CH ₂ ) _n -aryl, -( CH ₂ ) _n -heteroaryl or -(CH ₂ ) _n -heterocyclic group;

V in ULM-g is O, S or NR ₁ ;

R ₁ in ULM-g is the same as above;

R ¹ and R _1' in ULM-g each independently represent an H or a C ₁ -C ₃ alkyl group;

X ^R2' and X ^R3' in ULM-g are each independently optionally substituted -(CH ₂ ) _n -, -(CH ₂ ) _n -CH(X _v )=CH(X _v )- (cis or trans), -(CH ₂ ) _n -CH≡CH-, -(CH ₂ CH ₂ O)n- or C ₃ -C ₆ cycloalkyl group, where X _v represents H, halogen or C ₁ -C ₃ alkyl group, which is optionally substituted;

each m in ULM-g independently represents 0, 1, 2, 3, 4, 5, 6;

each m' in ULM-g is independently 0 or 1;

each n in ULM-g independently represents 0, 1, 2, 3, 4, 5, 6;

each n' in ULM-g is independently 0 or 1;

each u in ULM-g is independently a 0 or a 1;

each v in ULM-g is independently a 0 or a 1;

each w in ULM-g is independently a 0 or a 1; And

any one or more of R ^1' , R ^2' , R ^3' , X and X' in ULM-g is optionally modified to be covalently linked to a PTM group via a linker group when PTM is not ULM', or when PTM is is a ULM', any one or more of R ^1' , R ^2' , R ^3' , X and X' of each ULM and ULM' are optionally modified to be covalently linked to each other directly or through a linker group, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph.

[00221] In any of the aspects or embodiments described herein, ULM and, when present, ULM' are each independently a group according to chemical structure:

Where:

each of R ^1' , R ^2' and R ^3' in ULM-h is the same as above, and X is a C=O, C=S, -S(O) group or an S(O) ₂ group , more preferably a C=O group, and

any one or more of R ^1' , R ^2' and R ^3' in ULM-h are optionally modified to bind to a linker group to which a PTM group is further covalently linked when PTM is not ULM', or when PTM is ULM ', any one or more of R ^1' , R ^2' , R ^3' of each ULM and ULM' are optionally modified to be covalently linked to each other directly or through a linker group, or

a pharmaceutically acceptable salt, enantiomer, diastereomer, solvate or polymorph thereof.

[00222] In any of the aspects or embodiments described herein, ULM and, when present, ULM' each independently correspond to the chemical structure:

Where:

any one or more of R ^1' , R ^2' and R ^3' in ULM-I are optionally modified to bind to a linker group to which a PTM group is further covalently linked when PTM is not ULM', or when PTM is ULM ', any one or more of R ^1' , R ^2' , R ^3' of each ULM and ULM' are optionally modified to be covalently linked to each other directly or through a linker group, or

a pharmaceutically acceptable salt, enantiomer, diastereomer, solvate or polymorph thereof.

[00223] In further preferred aspects of the present invention, R ^1' in ULM-g - ULM-i is preferably a hydroxyl group or a group that can be metabolized to a hydroxyl or carboxyl group, such that the compound is a prodrug of the active compound. Typical preferred R ^1' groups include, for example, -(CH ₂ ) _n OH, (CH ₂ ) _n -O-(C ₁ -C ₆ )alkyl group, -(CH ₂ ) _n COOH, -(CH ₂ O) _n H, optionally substituted - (CH ₂ ) _n OC(O)-(C ₁ -C ₆ alkyl), or optionally substituted -(CH ₂ ) _n C(O)-O-(C ₁ -C ₆ alkyl), wherein n is 0 or 1. Wherein R ^1' is or contains a carboxylic acid group, a hydroxyl group, or an amine group, wherein the hydroxyl group, carboxylic acid group, or amine (each of which may be optionally substituted) may be further chemically modified to provide a covalent bond to the linker group to which the PTM group (including the ULM' group) is linked;

[00224] X and X', where present, in ULM-g and ULM-h are preferably a C=O, C=S, -S(O) group or a S(O) ₂ group, more preferably a C=O group;

[00225] R ^2' in ULM-g - ULM-i is preferably optionally substituted -NR ¹ -T-aryl (for example, optionally substituted NH-T-aryl or optionally substituted N(CH ₃ )-T-aryl), an optionally substituted -NR ¹ -T-heteroaryl group (for example, an optionally substituted NH-T-heteroaryl or an optionally substituted N(CH ₃ )-T-heteroaryl) or an optionally substituted -NR ¹ -T-heterocycle (for example, an optionally substituted NH- T-heterocycle or optionally substituted N(CH ₃ )-T-heterocycle), where R ¹ represents H or CH ₃ , preferably H and T represents an optionally substituted -(CH ₂ ) _n - group, where each of the methylene groups can be optionally substituted with one or two substituents, preferably selected from halogen, an amino acid side chain as otherwise described herein, or a C ₁ -C ₃ alkyl group, preferably one or two methyl groups, which may be optionally substituted; and n is 0-6, often 0, 1, 2 or 3, preferably 0 or 1. Alternatively, T may also be a -(CH ₂ O) _n group, -(OCH ₂ ) _n group, -( CH ₂ CH ₂ O) _n - group, -(OCH ₂ CH ₂ ) _n - group, all of which are optionally substituted.

[00226] Preferred aryl groups for R ^2' in ULM-g - ULM-i include optionally substituted phenyl or naphthyl groups, preferably phenyl groups wherein the phenyl or naphthyl group is linked to the PTM (including the ULM' group) by a linker group and/or optionally substituted with halogen (preferably F or Cl), amine, monoalkyl or dialkylamine (preferably dimethylamine), F, Cl, OH, COOH, C ₁ -C ₆ alkyl, preferably CH ₃ , CF ₃ , OMe, OCF ₃ , NO ₂ or a CN group (each of which may be substituted at the ortho, meta and/or para positions of the phenyl ring, preferably para), an optionally substituted phenyl group (the phenyl group itself is an optionally associated PTM group, including ULM', with a linker group), and/or a group optionally substituted by at least one of F, Cl, OH, COOH, CH ₃ , CF ₃ , OMe, OCF ₃ , NO ₂ or CN (at the ortho, meta and/or para positions of the phenyl rings, preferably para-), a naphthyl group which may be optionally substituted, an optionally substituted heteroaryl, preferably an optionally substituted isoxazole, including a methyl substituted isoxazole, an optionally substituted oxazole, including a methyl substituted oxazole, an optionally substituted thiazole, including a methyl substituted thiazole, an optionally substituted isothiazole, including methyl substituted isothiazole, optionally substituted pyrrole, including methyl substituted pyrrole, optionally substituted imidazole, including methylimidazole, optionally substituted benzimidazole or methoxybenzylimidazole, optionally substituted oximidazole or methyloximidazole, optionally substituted diazole group, including methyldiazole group, optionally substituted triazole group , including methyl-substituted triazole group , an optionally substituted pyridine group including a halo (preferably F) or methyl substituted pyridine group or an oxapyridine group (where the pyridine group is linked by a phenyl group via oxygen), optionally substituted furan, optionally substituted benzofuran, optionally substituted dihydrobenzofuran, optionally substituted indole, indolizine or azaindolysine (2, 3 or 4-azaindolysine), optionally substituted quinoline, optionally substituted group according to chemical structure:

Where:

S ^c in ULM-g - ULM-i represents CHR ^SS , NR ^URE , or O;

R ^HET in ULM-g - ULM-i is H, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted acetylene group -C≡CR _a where R _a represents H or C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl);

R ^SS in ULM-g - ULM-i is H, CN, NO ₂ , halogen (preferably F or Cl), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups), optionally substituted O-(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted -C(O)(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups);

R ^URE in ULM-g - ULM-i is H, C ₁ -C ₆ alkyl (preferably H or C ₁ -C ₃ alkyl) or -C(O)(C ₁ -C ₆ alkyl), each group optional substituted with one or two hydroxyl groups or up to three halogens, preferably fluorine groups, or an optionally substituted phenyl group, an optionally substituted heteroaryl or an optionally substituted heterocycle, preferably for example piperidine, morpholine, pyrrolidine, tetrahydrofuran);

R ^PRO in ULM-g - ULM-i is H, optionally substituted C ₁ -C ₆ alkyl or optionally substituted aryl (phenyl or naphthyl), heteroaryl or heterocyclic group selected from the group consisting of oxazole, isoxazole, thiazole, isothiazole , imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine, morpholine, quinoline, (each preferably substituted with a C ₁ -C ₃ alkyl group, preferably a methyl or halogen group , preferably F or Cl), benzofuran, indole, indolysine, azaindolysine; R ^PRO1 and R ^PRO2 in ULM-g - ULM-i each independently represent an H, optionally substituted C ₁ -C ₃ alkyl group or together form a keto group; And

each n in ULM-g - ULM-i is independently 0, 1, 2, 3, 4, 5 or 6 (preferably 0 or 1), or an optionally substituted heterocycle, preferably tetrahydrofuran, tetrahydrothiene, piperidine, piperazine or morpholine (each of the groups, when substituted, preferably substituted with methyl or halogen (F, Br, Cl), each of the groups may be optionally attached to a PTM group (including a ULM' group) via a linker group.

[00227] In certain preferred aspects in ULM-g - ULM-i represents

where R ^PRO and n in ULM-g - ULM-i are the same as above.

[00228] Preferred heteroaryl groups for R ^2' in ULM-g - ULM-i include optionally substituted quinoline (which may be attached to a pharmacophore or substituted on any carbon atom within the quinoline ring), optionally substituted indole, optionally substituted indolizine, optionally substituted azaindolysine, optionally substituted benzofuran, including optionally substituted benzofuran, optionally substituted isoxazole, optionally substituted thiazole, optionally substituted isothiazole, optionally substituted thiophene, optionally substituted pyridine (2-, 3 or 4-pyridine), optionally substituted imidazole, optionally substituted pyrrole, an optionally substituted diazole, an optionally substituted triazole, a tetrazole, an optionally substituted oximidazole, or a group according to the chemical structure:

Where:

S ^c in ULM-g - ULM-i represents CHR ^SS , NR ^URE , or O;

R ^HET in ULM-g - ULM-i is H, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), an optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or an optionally substituted acetylene group -C≡CR _a , where R _a in ULM-g is ULM- i represents H or a C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl);

R ^SS in ULM-g - ULM-i is H, CN, NO ₂ , halogen (preferably F or Cl), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups), optionally substituted O-(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted -C(O)(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups);

R ^URE in ULM-g - ULM-i is H, C ₁ -C ₆ alkyl (preferably H or C ₁ -C ₃ alkyl) or -C(O)(C ₁ -C ₆ alkyl), each of the groups of which , is optionally substituted with one or two hydroxyl groups or up to three halogens, preferably fluorine groups, or an optionally substituted heterocycle, for example piperidine, morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, piperidine, piperazine, each of which is optionally substituted, and

Y ^C in ULM-g - ULM-i is N or CR ^YC , where R ^YC is H, OH, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted one or two hydroxyl groups or up to three halogen groups (eg CF ₃ )), optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or an optionally substituted acetylene group -C≡ CR _a , where R _a represents H or a C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl), each of the groups optionally being linked to a PTM group (including a ULM' group) via a linker group.

[00229] Preferred heterocyclic groups for R ^2' in ULM-g - ULM-i include tetrahydrofuran, tetrahydrothiene, tetrahydroquinoline, piperidine, piperazine, pyrrolidine, morpholine, ocean or tian, each of which groups may be optionally substituted, or a group according to with chemical structure:

Preferably, group,

Where:

R ^PRO in ULM-g - ULM-i represents H, an optionally substituted C ₁ -C ₆ alkyl or an optionally substituted aryl, heteroaryl or heterocyclyl group;

R ^PRO1 and R ^PRO2 in ULM-g - ULM-i each independently represent an H, optionally substituted C ₁ -C ₃ alkyl group or together form a keto group and

each n in ULM-g - ULM-i independently represents 0, 1, 2, 3, 4, 5 or 6 (often 0 or 1), each of the groups may optionally be associated with a PTM group (including a ULM' group) via linker group.

[00230] Preferred R ^2' substituents in ULM-g - ULM-i also include, in particular (and without limitation to the specific compound disclosed) R ^2' substituents that are found in certain compounds disclosed herein (which include specific compounds that disclosed in the specification and figures presented herein). Each of these R ^2' substituents can be used in combination with any number of R ^3' substituents, which are also disclosed herein.

[00231] R ^3' in ULM-g - ULM-i is preferably optionally substituted -T-aryl, optionally substituted-T-heteroaryl, optionally substituted -T-heterocyclyl, optionally substituted-NR ¹ -T-aryl (e.g. optionally substituted NH-T-aryl, optionally substituted N(CH ₃ )-T-aryl or optionally substituted N(C ₁ -C ₃ alkyl)-T-aryl), optionally substituted -NR ¹ -T-heteroaryl (e.g., optionally substituted NH-T-heteroaryl, optionally substituted N(CH ₃ )-T-heteroaryl, or optionally substituted N(C ₁ -C ₃ alkyl)-T-heteroaryl), or optionally substituted -NR ¹ -T-heterocyclyl (for example, optionally substituted NH-T-heterocyclyl, optionally substituted N(CH ₃ )-T-heterocyclyl or optionally substituted N(C ₁ -C ₃ alkyl)-T-heterocyclyl), where R ¹ represents H or C ₁ -C ₃ alkyl group, preferably H or CH ₃ , T optionally substituted -(CH ₂ ) _n - group, where each of the methylene groups may be optionally substituted with one or two substituents, preferably selected from halogen, a C ₁ -C ₃ alkyl group or an amino acid side chain , as otherwise described herein, preferably methyl, which may be optionally substituted; and n is 0-6, often 0, 1, 2 or 3, preferably 0 or 1. Alternatively, T may also be a -(CH ₂ O) _n group, -(OCH ₂ ) _n group, -( CH ₂ CH ₂ O) _n - group, -(OCH ₂ CH ₂ ) _n - group, each of the groups is optionally substituted.

[00232] Preferred aryl groups for R ^3' in ULM-g to ULM-i include optionally substituted phenyl or naphthyl groups, preferably phenyl groups wherein the phenyl or naphthyl group is optionally linked to a PTM group (including a ULM' group) via a linker group and /or optionally substituted with a halogen (preferably F or Cl), amine, monoalkyl or dialkylamine (preferably dimethylamine), amido group (preferably -(CH ₂ ) _m - NR ₁ C(O)R ₂ group, where m, R ₁ and R ₂ are the same as above), halogen (often F or Cl), OH, CH ₃ , CF ₃ , OMe, OCF ₃ , NO ₂ , CN or a S(O) _{2 RS} group (R _S represents a C ₁ -C ₆ alkyl group, optionally substituted aryl, heteroaryl, or heterocyclic group, or - (CH ₂ ) _m NR ₁ R ₂ group), each of which may be substituted at the ortho, meta and/or para positions phenyl ring, preferably para-), or aryl (preferably phenyl), heteroaryl or heterocycle. Preferably, said substituted phenyl group is an optionally substituted phenyl group (i.e., the substituted phenyl group is preferably substituted by at least one of F, Cl, OH, SH, COOH, CH ₃ , CF ₃ , OMe, OCF ₃ , NO ₂ , CN or a linker group to which is attached a PTM group (including a ULM' group), where the substitution occurs at the ortho, meta and/or para positions of the phenyl ring, preferably para), a naphthyl group which may be optionally substituted, Including, as described above, not necessarily replaced by heteroaril (preferably optionally replaced by the Izoxazole, including methyl -stained Izoxazole, optionally replaced oxazole, including methyl -recessed oxazole, not necessarily substituted thiazol, including methyl -substituted thiazole, optionally noticeable pyrrol, including methyl -designed pyrrhum, unsubstantially noticeable image including methylimidazole, benzylimidazole or methoxybenzylimidazole, oximidazole or methyloximidazole, an optionally substituted diazole group including a methyldiazole group, an optionally substituted triazole group including a methyl substituted triazole group, a pyridine group including a halo-preferably F) or a methyl substituted pyridine group or an oxapyridine group (wherein the pyridine group is bonded to phenyl group via oxygen) or an optionally substituted heterocycle (tetrahydrofuran, tetrahydrothiophene, pyrrolidine, piperidine, morpholine, piperazine, tetrahydroquinoline, ocean or tian). Each aryl, heteroaryl or heterocyclic group may be optionally linked to a PTM group (including a ULM' group) via a linker group.

[00233] Preferred heteroaryl groups for R ^3' in ULM-g - ULM-i include optionally substituted quinoline (which may be attached to a pharmacophore or substituted on any carbon atom on the quinoline ring), optionally substituted indole (including dihydroindole), optionally substituted indolizine, optionally substituted azaindolizine (2, 3 or 4-azaindolizine), optionally substituted benzimidazole, benzodiazole, benzoxofuran, optionally substituted imidazole, optionally substituted isoxazole, optionally substituted oxazole (preferably methyl substituted), optionally substituted diazole, optionally substituted triazole, tetrazole, optional substituted benzofuran, optionally substituted thiophene, optionally substituted thiazole (preferably substituted with methyl and/or thiol), optionally substituted isothiazole, optionally substituted triazole (preferably 1,2,3-triazole substituted with a methyl group, triisopropylsilyl group, optionally substituted with -(CH ₂ ) _m OC ₁ -C ₆ alkyl group or an optionally substituted -(CH ₂ ) _m C(O)-OC ₁ -C ₆ alkyl group), optionally substituted pyridine (2-, 3- or 4-pyridine) or a group according to with chemical structure:

Where:

S ^c in ULM-g - ULM-i represents CHR ^SS , NR ^URE , or O;

R ^HET in ULM-g - ULM-i is H, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted acetylene group -C≡CR _a where R _a represents H or C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl);

R ^SS in ULM-g - ULM-i is H, CN, NO ₂ , halogen (preferably F or Cl), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups), optionally substituted O-(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted -C(O)(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups);

R ^URE in ULM-g - ULM-i is H, C ₁ -C ₆ alkyl (preferably H or C ₁ -C ₃ alkyl) or -C(O)(C ₁ -C ₆ alkyl), each of the groups of which , optionally substituted with one or two hydroxyl groups or up to three halogens, preferably fluorine groups, or optionally

a substituted heterocycle, for example piperidine, morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, piperidine, piperazine, each of which is optionally substituted, and

Y ^C in ULM-g - ULM-i is N or CR ^YC , where R ^YC is H, OH, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), an optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or an optionally substituted acetylene group -C≡CR _a where R _a represents H or a C ₁ -C ₆ alkyl group (preferably a C ₁ -C ₃ alkyl group) Each of these heteroaryl groups may optionally be linked/attached to a PTM group (including a ULM' group) via a linker group.

[00234] Preferred heterocyclic groups for R ^3' in ULM-g - ULM-i include tetrahydroquinoline, piperidine, piperazine, pyrrolidine, morpholine, tetrahydrofuran, tetrahydrothiophene, ocean and tian, each of which groups may be optionally substituted, or a group in according to chemical structure:

, preferably

group,

Where:

R ^PRO in ULM-g - ULM-i is H, optionally substituted C ₁ -C ₆ alkyl or optionally substituted aryl (phenyl or naphthyl), heteroaryl or heterocyclic group selected from the group consisting of oxazole, isoxazole, thiazole, isothiazole , imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine, morpholine, quinoline, (each preferably substituted with a C ₁ -C ₃ alkyl group, preferably a methyl or halogen group , preferably F or Cl), benzofuran, indole, indolysine, azaindolysine; R ^PRO1 and R ^PRO2 in ULM-g - ULM-i each independently represent an H, optionally substituted C ₁ -C ₃ alkyl group or together form a keto group and

each t in ULM-g - ULM-i is 0, 1, 2, 3, 4, 5 or 6 (preferably 0 or 1), where each of said heterocyclic groups may be optionally associated/attached to a PTM group (including ULM ' group) through a linker group.

[00235] Preferred R ^3' substituents in ULM-g - ULM-i also include, in particular (and without limitation to the specific compound disclosed) R ^3' substituents that are found in certain compounds disclosed herein (which include specific compounds that disclosed in the specification and figures attached herein). Each of these R ^3' substituents may be used in combination with any number of R ^2' substituents also disclosed herein.

[00236] In certain alternative preferred embodiments, R ^2' in ULM-g - ULM-i is an optionally substituted -NR ₁ -X ^R2' alkyl group, -NR ₁ -XR ^2' aryl group; optionally substituted -NR ₁ -X ^R2' -NO, optionally substituted -NR ₁ -XR ^2' -aryl-NO or optionally substituted -NR ₁ -XR ^2' -NO-aryl, where:

R ₁ in ULM-g - ULM-i represents H or a C ₁ -C ₃ alkyl group (preferably H);

X ^R2' in ULM-g - ULM-i is optionally substituted -CH ₂ ) _n -, -CH ₂ ) _n -CH(X _v )=CH(X _v )- (cis or trans), -(CH ₂ ) _n -CH≡CH-, -(CH ₂ CH ₂ O) _n - or C ₃ -C ₆ cycloalkyl group; And

X _v in ULM-g - ULM-i represents H, halogen or a C ₁ -C ₃ alkyl group, which is optionally substituted with one or two hydroxyl groups or up to three halogen groups;

The alkyl in ULM-g - ULM-i is an optionally substituted C ₁ -C ₁₀ alkyl (preferably C ₁ -C ₆ alkyl) group (in certain preferred embodiments, the alkyl group is terminated with a halogen group, often Cl or Br) ;

The aryl in ULM-g - ULM-i is an optionally substituted phenyl or naphthyl group (preferably a phenyl group); and NO in ULM-g - ULM-i is an optionally substituted oxazole, isoxazole, thiazole, isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine, morpholine, benzofuran, indole, indolizine, azaindolizine, quinoline (when substituted, each is preferably substituted with a C ₁ -C ₃ alkyl group, preferably a methyl or halogen group, preferably F or Cl) or group according to the chemical structure:

S ^c in ULM-g - ULM-i represents CHR ^SS , NR ^URE , or O;

R ^HET in ULM-g - ULM-i is H, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted acetylene group -C≡CR _a where R _a represents H or C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl);

R ^SS in ULM-g - ULM-i is H, CN, NO ₂ , halogen (preferably F or Cl), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups), optionally substituted O-(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted -C(O)(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups);

R ^URE in ULM-g - ULM-i is H, C ₁ -C ₆ alkyl (preferably H or C ₁ -C ₃ alkyl) or -C(O)(C ₁ -C ₆ alkyl), each of the groups of which , optionally substituted with one or two hydroxyl groups or up to three halogens, preferably fluorine groups, or an optionally substituted heterocycle, for example piperidine, morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, piperidine, piperazine, each of which is optionally substituted;

Y ^C in ULM-g - ULM-i is N or CR ^YC , where R ^YC is H, OH, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), an optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or an optionally substituted acetylene group -C≡CR _a , where R _a represents H or a C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl);

R ^PRO in ULM-g - ULM-i is H, optionally substituted C ₁ -C ₆ alkyl or optionally substituted aryl (phenyl or naphthyl), heteroaryl or heterocyclic group selected from the group consisting of oxazole, isoxazole, thiazole, isothiazole , imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine, morpholine, quinoline, (each preferably substituted with a C ₁ -C ₃ alkyl group, preferably a methyl or halogen group , preferably F or Cl), benzofuran, indole, indolysine, azaindolysine;

R ^PRO1 and R ^PRO2 in ULM-g - ULM-i each independently represent an H, optionally substituted C ₁ -C ₃ alkyl group or together form a keto group and

each n in ULM-g - ULM-i is independently 0, 1, 2, 3, 4, 5 or 6 (preferably 0 or 1).

[00237] Each of these groups may optionally be linked/attached to a PTM group (including a ULM' group) via a linker group.

[00238] In certain alternative preferred embodiments of the compound of the present invention, R ^3' in ULM-g - ULM-i is optionally substituted -(CH ₂ ) _n -(V)n-(CH ₂ ) _n -(V)nR ^S3' group, optionally substituted -(CH ₂ ) _n -N(R _1' )(C=O) _m' -(V) _n' -R ^S3' group, optionally substituted -X ^R3' -alkyl group, optionally substituted -X ^R3' -aryl group; an optionally substituted -X ^R3' -NO group, an optionally substituted -X ^R3' -aryl-NO group or an optionally substituted -X ^R3' -NO-aryl group, where:

R ^S3' represents an optionally substituted alkyl group (C ₁ -C ₁₀ , preferably C ₁ -C ₆ alkyl), an optionally substituted aryl group or a NO group;

R _1' represents H or a C ₁ -C ₃ alkyl group (preferably H);

V represents O, S or NR _1' ;

X ^R3' represents -(CH ₂ ) _n -, -(CH ₂ CH ₂ O) _n -, -CH ₂ ) _n -CH(X _v )=CH(X _v )- (cis or trans), -CH ₂ ) _n -CH≡CH- or C ₃ -C ₆ cycloalkyl group, all optionally substituted;

X _v represents H, halogen or a C ₁ -C ₃ alkyl group, which is optionally substituted with one or two hydroxyl groups or up to three halogen groups;

Alkyl is an optionally substituted C ₁ -C ₁₀ alkyl (preferably C ₁ -C ₆ alkyl) group (in certain preferred embodiments, the alkyl group is terminated with a halogen group, often Cl or Br);

Aryl represents an optionally substituted phenyl or naphthyl group (preferably a phenyl group); and NO is an optionally substituted oxazole, isoxazole, thiazole, isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine, morpholine, benzofuran, indole, indolizine, azaindolysine, quinoline (when substituted, each is preferably substituted with a C ₁ -C ₃ alkyl group, preferably a methyl or halogen group, preferably F or Cl) or group according to the chemical structure:

S ^c in ULM-g - ULM-i represents CHR ^SS , NR ^URE , or O;

R ^HET in ULM-g - ULM-i is H, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted acetylene group -C≡CR _a where R _a represents H or C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl);

R ^SS in ULM-g - ULM-i is H, CN, NO ₂ , halogen (preferably F or Cl), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups), optionally substituted O-(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted -C(O)(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups);

R ^URE in ULM-g - ULM-i is H, C ₁ -C ₆ alkyl (preferably H or C ₁ -C ₃ alkyl) or -C(O)(C ₀ -C ₆ alkyl), each of the groups of which , optionally substituted with one or two hydroxyl groups or up to three halogens, preferably fluorine groups, or an optionally substituted heterocycle, for example piperidine, morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, piperidine, piperazine, each of which is optionally substituted;

Y ^C in ULM-g - ULM-i is N or CR ^YC , where R ^YC is H, OH, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), an optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or an optionally substituted acetylene group -C≡CR _a , where R _a represents H or a C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl);

R ^PRO in ULM-g - ULM-i is H, optionally substituted C ₁ -C ₆ alkyl or optionally substituted aryl (phenyl or naphthyl), heteroaryl or heterocyclic group selected from the group consisting of oxazole, isoxazole, thiazole, isothiazole , imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine, morpholine, quinoline, (each preferably substituted with a C ₁ -C ₃ alkyl group, preferably a methyl or halogen group , preferably F or Cl), benzofuran, indole, indolysine, azaindolysine; R ^PRO1 and R ^PRO2 in ULM-g - ULM-i each independently represent an H, optionally substituted C ₁ -C ₃ alkyl group or together form a keto group;

each n in ULM-g - ULM-i is independently 0, 1, 2, 3, 4, 5 or 6 (preferably 0 or 1);

each m' in ULM-g - ULM-i represents 0 or 1; And

each n' in ULM-g - ULM-i represents 0 or 1;

wherein each of said compounds, preferably on an alkyl, aryl or Het group, is optionally linked/attached to a PTM group (including a ULM' group) via a linker.

[00239] In alternative embodiments, R ^3' , in ULM-g - ULM-i is -(CH ₂ ) _n -aryl, -(CH ₂ CH ₂ O) _n -aryl, -(CH ₂ ) _n -NO or - (CH ₂ CH ₂ O) _n -NO,

Where:

said aryl in ULM-g - ULM-i is phenyl, which is optionally substituted with one or two substituents, wherein said substituent(s) is preferably selected from -(CH ₂ ) _n OH, C ₁ -C ₆ alkyl, which is additionally optionally substituted with CN, halogen (up to three halogen groups), OH, -(CH ₂ ) _n O(C ₁ -C ₆ )alkyl, amine, mono- or di-(C ₁ -C ₆ alkyl) amine, where alkyl the amine group is optionally substituted with 1 or 2 hydroxyl groups or up to three halogen (preferably F, Cl) groups, or

the specified aryl group in ULM-g - ULM-i is replaced by -(CH ₂ ) _n OH, -(CH ₂ ) _n -O-(C ₁ -C ₆ )alkyl, -(CH ₂ ) _n -O-(CH ₂ ) _n -(C ₁ -C ₆ ) alkyl, -(CH ₂ ) _n -C(O)(C ₀ -C ₆ ) alkyl, -(CH ₂ ) _n -C(O)O(C ₀ -C ₆ )alkyl, -(CH ₂ ) _n -OC(O)(C ₀ -C ₆ )alkyl, amine, mono- or di-(C ₁ -C ₆ alkyl) amine, where the alkyl group of the amine is optionally replaced by 1 or 2 hydroxyl groups or up to three halogen (preferably F, Cl) groups, CN, NO ₂ , optionally substituted -(CH ₂ ) _n -(V) _m' -CH ₂ ) _n -(V) _m' -(C ₁ -C ₆ )alkyl group, -(V) _m' -(CH ₂ CH ₂ O)nR ^PEG group, where V represents O, S or NR _1' , R _1' represents H or C ₁ -C ₃ alkyl group (preferably H) and R ^PEG represents H or a C ₁ -C ₆ alkyl group that is optionally substituted (including optional substitution with a carboxyl group), or

said aryl group in ULM-g - ULM-i is optionally substituted with a heterocycle, including heteroaryl selected from the group consisting of oxazole, isoxazole, thiazole, isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine, morpholine, quinoline, benzofuran, indole, indolysine, azaindolysine (when each substituted is preferably substituted with a C ₁ -C ₃ alkyl group, preferably a methyl or halogen group, preferably F or Cl), or a group according to chemical structure:

S ^c in ULM-g - ULM-i represents CHR ^SS , NR ^URE , or O;

R ^HET in ULM-g - ULM-i is H, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted acetylene group -C≡CR _a where R _a represents H or C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl);

R ^SS in ULM-g - ULM-i is H, CN, NO ₂ , halogen (preferably F or Cl), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups), optionally substituted 0-(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted -C(O)(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups);

R ^URE in ULM-g - ULM-i is H, C ₁ -C ₆ alkyl (preferably H or C ₁ -C ₃ alkyl) or -C(O)(C ₀ -C ₆ alkyl), each of the groups of which optionally substituted with one or two hydroxyl groups or up to three halogens, preferably fluorine groups, or an optionally substituted heterocycle, for example piperidine, morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, piperidine, piperazine, each of which is optionally substituted;

Y ^C in ULM-g - ULM-i is N or CR ^YC , where R ^YC is H, OH, CN, N02, halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), an optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or an optionally substituted acetylene group -C≡CR _a where R _a represents H or a C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl);

R ^PRO in ULM-g - ULM-i is H, optionally substituted C ₁ -C ₆ alkyl or optionally substituted aryl (phenyl or naphthyl), heteroaryl or heterocyclic group selected from the group consisting of oxazole, isoxazole, thiazole, isothiazole , imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine, morpholine, quinoline, (each preferably substituted with a C ₁ -C ₃ alkyl group, preferably a methyl or halogen group , preferably F or Cl), benzofuran, indole, indolysine, azaindolysine;

R ^PRO1 and R ^PRO2 in ULM-g - ULM-i each independently represent an H, optionally substituted C ₁ -C ₃ alkyl group or together form a keto group;

NO in ULM-g - ULM-i is preferably oxazole, isoxazole, thiazole, isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine, morpholine, quinoline (each preferably substituted with a C ₁ -C ₃ alkyl group, preferably a methyl or halogen group, preferably F or Cl), benzofuran, indole, indolizine, azaindolizine, or group according to the chemical structure:

S ^c in ULM-g - ULM-i represents CHR ^SS , NR ^URE , or O;

R ^HET in ULM-g - ULM-i is H, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted acetylene group -C≡CR _a where R _a represents H or C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl); R ^SS in ULM-g - ULM-i represents H, CN, NO ₂ , halogen (preferably F or Cl), optionally substituted C ₁ -C ₆ alkyl (preferably substituted with one or two hydroxyl groups or up to three halogen groups), optionally substituted O-(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or optionally substituted -C(O)(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups);

R ^URE in ULM-g - ULM-i is H, C ₁ -C ₆ alkyl (preferably H or C ₁ -C ₃ alkyl) or -C(O)(C ₀ -C ₆ alkyl), each group optional substituted with one or two hydroxyl groups or up to three halogens, preferably fluorine groups, or an optionally substituted heterocycle, for example piperidine, morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, piperidine, piperazine, each of which is optionally substituted;

Y ^C in ULM-g - ULM-i is N or CR ^YC , where R ^YC is H, OH, CN, NO ₂ , halogen (preferably Cl or F), optionally substituted C ₁ -C ₆ alkyl (preferably substituted one or two hydroxyl groups or up to three halogen groups (eg CF ₃ ), an optionally substituted O(C ₁ -C ₆ alkyl) (preferably substituted with one or two hydroxyl groups or up to three halogen groups) or an optionally substituted acetylene group -C≡CR _a , where R _a represents H or a C ₁ -C ₆ alkyl group (preferably C ₁ -C ₃ alkyl);

R ^PRO in ULM-g - ULM-i is H, optionally substituted C ₁ -C ₆ alkyl or optionally substituted aryl, heteroaryl or heterocyclic group;

R ^PRO1 and R ^PRO2 in ULM-g - ULM-i each independently represent an H, optionally substituted C ₁ -C ₃ alkyl group or together form a keto group;

each m' in ULM-g - ULM-i is independently 0 or 1; And

each n in ULM-g - ULM-i is independently 0, 1, 2, 3, 4, 5 or 6 (preferably 0 or 1),

wherein each of said compounds, preferably on said aryl or NO groups, is optionally linked/attached to a PTM group (including a ULM' group) via a linker group.

[00240] In still further embodiments, preferred compounds include those corresponding to the chemical structure:

Where:

R ^1' in ULM-i represents OH or a group that is metabolized in the patient or subject to OH;

R ^2' in ULM-i is -NH-CH ₂ -aryl-NET (preferably phenyl linked directly to a methyl-substituted thiazole);

R ^3' in ULM-i is a -CHR ^CR3' -NH-C(O)-R ^3P1 group or a -CHR ^CR3' -R ^3P2 group;

R ^CR3' in ULM-i represents a C ₁ -C ₄ alkyl group, preferably methyl, isopropyl or tert-butyl;

R ^3P1 in ULM-i is a C ₁ -C ₃ alkyl (preferably methyl), optionally substituted oxetane group (preferably methyl, -(CH ₂ ) _n OCH ₃ group, where n is 1 or 2 (preferably 2), or a group (wherein the ethyl ester group is preferably meta-substituted with a phenyl moiety), a morpholino group (linked to the carbonyl at the 2nd or 3rd position);

R ^3P2 in ULM-i represents group;

aryl in ULM-i is phenyl;

NO in ULM-i is an optionally substituted thiazole or isothiazole; And

R ^HET in ULM-i represents H or a halogen group (preferably H);

or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof, wherein each of these compounds is optionally linked to a PTM group (including a ULM' group) via a linker group.

[00241] In certain aspects, the bifunctional compounds contain an E3 ubiquitin ligase binding moiety (ULM), wherein ULM is a group according to the chemical structure:

Where:

each R ₅ and R ₆ in ULM-j independently represents OH, SH or optionally substituted alkyl, or R ₅ , R ₆ , and the carbon atom to which they are attached forms a carbonyl;

R ⁷ in ULM-j is H or optionally substituted alkyl;

E in ULM-j represents a bond, C=O, or C=S;

G in ULM-j represents a bond, optionally substituted alkyl, -COOH or C=J;

J in ULM-j represents O or NR ₈ ;

R ₈ in ULM-j is H, CN, optionally substituted alkyl or optionally substituted alkoxy;

M in ULM-j is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, or

R ₉ and R ₁₀ in ULM-j are each independently H; optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted hydroxyalkyl, optionally substituted thioalkyl, disulfide-linked ULM, optionally substituted heteroaryl or haloalkyl; or R ₉ , R ₁₀ , and the carbon atom to which they are attached form an optionally substituted cycloalkyl;

R ₁₁ in ULM-j is an optionally substituted heterocycle, optionally substituted alkoxy, optionally substituted heteroaryl, optionally substituted aryl, or

R ₁₂ in ULM-j represents H or optionally substituted alkyl;

R ₁₃ in ULM-j is H, optionally substituted alkyl, optionally substituted alkylcarbonyl, optionally substituted (cycloalkyl)alkylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted arylcarbonyl, optionally substituted (heterocyclyl)carbonyl, or optionally substituted aralkyl; optionally substituted (oxoalkyl)carbamate,

each R ₁₄ in ULM-j independently represents H, haloalkyl, optionally substituted cycloalkyl, optionally substituted alkyl, or optionally substituted heterocycloalkyl;

R ₁₅ in ULM-j is H, CN, optionally substituted heteroaryl, haloalkyl, optionally substituted aryl, optionally substituted alkoxy or optionally substituted heterocyclyl;

each R ₁₆ in ULM-j independently represents halogen, optionally substituted alkyl, optionally substituted haloalkyl, CN or optionally substituted haloalkoxy;

each R ₂₅ in ULM-j independently represents H or optionally substituted alkyl; or both R ₂₅ groups can be taken together to form an oxo or optionally substituted cycloalkyl group;

R ₂₃ in ULM-j is H or OH;

Z ₁ , Z ₂ , Z ₃ , and Z ₄ in ULM-j are independently C or N; And

o in ULM-j is 0, 1, 2, 3 or 4, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof.

[00242] In certain embodiments, wherein G in ULM-j is C=J, J is O, _R7 is H, each _R14 is H, and o is 0.

[00243] In certain embodiments, wherein G in ULM-j is C=J, J is O, R ₇ is H, each R ₁₄ is H, R ₁₅ is optionally substituted heteroaryl, and o is 0. In other cases, E represents C=O and M represents

[00244] In certain embodiments, where E in ULM-j is C=O, R ₁₁ is an optionally substituted heterocycle, or and M represents

[00245] In certain embodiments, where E in ULM-j is C=O, M is , and R ₁₁ represents or each R ₁₈ independently represents halogen, optionally substituted alkoxy, cyano, optionally substituted alkyl, haloalkyl or haloalkoxy; and p is 0, 1, 2, 3 or 4.

[00246] In certain embodiments, ULMs and where present, ULM', are each independently a group according to chemical structure:

Where:

G in ULM-k represents C=J, J represents O;

R ₇ in ULM-k is H;

each R ₁₄ in ULM-k represents H;

o in ULM-k represents 0;

R ₁₅ in ULM-k represents And

R ₁₇ in ULM-k is H, halogen, optionally substituted cycloalkyl, optionally substituted alkyl, optionally substituted alkenyl and haloalkyl.

[00247] In other cases, R ₁₇ in ULM-k is alkyl (eg, methyl) or cycloalkyl (eg, cyclopropyl).

[00248] In other embodiments, ULM and where present, ULM' are each independently a group according to chemical structure:

Where:

G in ULM-k represents C=J, J represents O;

R ₇ in ULM-k is H;

each R ₁₄ in ULM-k represents H;

o in ULM-k represents 0; And

R ₁₅ in ULM-k is selected from the group consisting of:

where R ₃₀ in ULM-k represents H or optionally substituted alkyl.

[00249] In other embodiments, ULM and where present, ULM' are each independently a group according to chemical structure:

Where:

E in ULM-k represents C=O;

The M in ULM-k represents ; And

R ₁₁ in ULM-k is selected from the group consisting of:

[00250] In still other embodiments, a compound of a chemical structure,

where E in ULM-k represents C=O;

R ₁₁ in ULM-k represents ; And

The M in ULM-k represents ;

q in ULM-k represents 1 or 2;

R ₂₀ in ULM-k is H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, or

R ₂₁ in ULM-k represents H or optionally substituted alkyl; And

R ₂₂ in ULM-k is H, optionally substituted alkyl, optionally substituted alkoxy or haloalkyl.

[00251] In any embodiment described herein, R ₁₁ in ULM-j or ULM-k is selected from the group consisting of:

[00252] In certain embodiments, R ₁₁ in ULM-j or ULM-k is selected from the group consisting of:

[00253] In certain embodiments, ULM (or when ULM' is present) is a group according to the chemical structure:

Where:

X in ULM-I represents O or S;

Y in ULM-1 is H, methyl or ethyl;

R ₁₇ in ULM-1 is H, methyl, ethyl, hydroxymethyl or cyclopropyl;

M in ULM-1 is optionally substituted aryl, optionally substituted heteroaryl, or ;

R ₉ in ULM-1 is H;

R ₁₀ in ULM-1 is H, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted hydroxyalkyl, optionally substituted thioalkyl or cycloalkyl;

R ₁₁ in ULM-1 is an optionally substituted heteroaromatic group, an optionally substituted heterocycle, an optionally substituted aryl, or ;

R ₁₂ in ULM-1 represents H or optionally substituted alkyl; And

R ₁₃ in ULM-1 is H, optionally substituted alkyl, optionally substituted alkylcarbonyl, optionally substituted (cycloalkyl)alkylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted arylcarbonyl, optionally substituted (heterocyclyl)carbonyl, or optionally substituted aralkyl; optionally substituted (oxoalkyl)carbamate.

[00254] In some embodiments, ULM and where present, ULM' are each independently a group according to chemical structure:

Where:

Y in ULM-m is H, methyl or ethyl

R ₉ in ULM-m is H;

R ₁₀ is isopropyl, t-butyl, sec-butyl, cyclopentyl or cyclohexyl;

R ₁₁ in ULM-m represents an optionally substituted amide, an optionally substituted isoindolinone, an optionally substituted isoxazole, an optionally substituted heterocycle.

[00255] In other preferred embodiments, ULM and where present, ULM', are each independently a group according to chemical structure:

Where:

R ₁₇ in ULM-n is methyl, ethyl or cyclopropyl; And

R ₉ , R ₁₀ , and R ₁₁ in ULM-n are as defined above. In other cases, R ₉ represents H; And

R ₁₀ in ULM-n is H, alkyl or cycloalkyl (preferably isopropyl, t-butyl, sec-butyl, cyclopentyl or cyclohexyl).

[00256] In any of the aspects or embodiments described herein, ULM (or when present, ULM'), as described herein, may be a pharmaceutically acceptable salt, enantiomer, diastereomer, solvate, or polymorph thereof. Additionally, in any of the aspects or embodiments described herein, the ULM (or when present, ULM') as described herein may be linked to the PTM directly through a linkage or via a chemical linker.

[00257] In certain aspects of the present invention, the ULM fragment is selected from the group consisting of:

And

wherein the VLM may be linked to the PTM via a linker as described herein at any appropriate location including, for example, an aryl, heteroaryl, phenyl or phenyl indole group, optionally via any suitable functional group such as an amine, an ester, an ether , alkyl or alkoxy.

[00258] Typical linkers

[00259] In certain embodiments, the compounds as described herein include one or more PTMs chemically linked to or linked to one or more ULMs (e.g., at least one of CLM, VLM, MLM, ILM, or a combination thereof) via chemical linker (L). In certain embodiments, a linker group L is a group containing one or more covalently linked structural elements (e.g., -A ^L ₁ ...(A ^L ) _q - or -(A ^L ) _q -), wherein A ^L ₁ is a group , associated with the RTM, a (A ^L ) _q represents the group associated with the ULM.

[00260] In any aspect or embodiment described herein, the linker group L is a chemical linker bond or group represented by the formula -(A ^L ) _q -, where A ^L is a chemical moiety and q is an integer of 1-100, and wherein A ^L is covalently linked to PTM and ULM, and is proposed to efficiently bind PTM to a target protein and ULM to an E3 ubiquitin ligase, resulting in targeted ubiquitination of the protein.

[00261] In certain embodiments, the linker group L is -(A ^L ) _q -, where:

(A ^L ) _q represents a group that is associated with at least one of a ULM (such as CLM or VLM), a PTM moiety, or a combination thereof; linker q is an integer greater than or equal to 1;

each A ^L is independently selected from the group consisting of a link, CR ^L1 R ^L2 , O, S, SO, SO ₂ , NR ^L3 , SO ₂ NR ^L3 , SONR ^L3 , CONR ^L3 , NR ^L3 CONR ^L4 , NR ^L3 SO ₂ NR ^L4 , CO, CR ^L1 =CR ^L2 , C≡C, SiR ^L1 R ^L2 , P(O)R ^L1 , P(O)OR ^L1 , NR ^L3 C(=NCN)NR ^L4 , NR ^L3 C(=NCN) , NR ^L3 C(=CNO ₂ )NR ^L4 , C _3-11 cycloalkyl, optionally substituted with 0-6 R ^L1 and/or R ^L2 groups, C _5-13 spirocycloalkyl, optionally substituted with 0-9 R ^L1 and/or R ^L2 groups, C _3-11 heterocyclyl, optionally substituted with 0-6 R ^L1 and/or R ^L2 groups, C _5-13 spiroheterocycloalkyl, optionally substituted with 0-8 R ^L1 and/or R ^L2 groups, aryl, optionally substituted with 0-6 R ^L1 and/or R ^L2 groups, heteroaryl, optionally substituted with 0-6 R ^L1 and/or R ^L2 groups, wherein R ^L1 or R ^L2 are each independently optionally linked to other groups to form a cycloalkyl and/or heterocyclyl moiety, optionally substituted with 0 -4 R ^L5 groups; And

R ^L1 , R ^L2 , R ^L3 , R ^L4 and R ^L5 are each independently H, halogen, C _1-8 alkyl, OC _1-8 alkyl, SC _1-8 alkyl, NHC _1-8 alkyl, N(C _{1 -8} alkyl) ₂ , C _3-11 cycloalkyl, aryl, heteroaryl, C _3-11 heterocyclyl, OS _1-8 cycloalkyl, SC _1-8 cycloalkyl, NHC _1-8 cycloalkyl, N(C _1-8 cycloalkyl) ₂ , N(C _1-8 cycloalkyl)(C _1-8 alkyl), OH, NH ₂ , SH, SO ₂ C _1-8 alkyl, P(O)(OS _1-8 alkyl)(C _1-8 alkyl), P(O)(OS _1-8 alkyl) ₂ , CC-C _1-8 alkyl, CCH, CH=CH(C _1-8 alkyl), C(C _1-8 alkyl)=CH(C _1-8 alkyl ), C(C _1-8 alkyl)=C(C _1-8 alkyl) ₂ , Si(OH) ₃ , Si(C _1-8 alkyl) ₃ , Si(OH)(C _1-8 alkyl) ₂ , COC _1-8 alkyl, CO ₂ H, halogen, CN, CF ₃ , CHF ₂ , CH ₂ F, NO ₂ , SF ₅ , SO ₂ NHC _1-8 alkyl, SO ₂ N(C _1-8 alkyl) ₂ , SONHC _1-8 alkyl, SON(C _1-8 alkyl) ₂ , CONHC _1-8 alkyl, CON(C _1-8 alkyl) ₂ , N(C _1-8 alkyl)CONH(C _1-8 alkyl), N (C _1-8 alkyl)CON(C _1-8 alkyl) ₂ , NHCONH(C _1-8 alkyl), NHCON(C _1-8 alkyl) ₂ , NHCONH ₂ , N(C _1-8 alkyl)SO ₂ NH (C _1-8 alkyl), N(C _1-8 alkyl) SO ₂ N(C _1-8 alkyl) ₂ , NH SO ₂ NH(C _1-8 alkyl), NH SO ₂ N(C _1-8 alkyl ) ₂ , NH SO ₂ NH ₂ .

[00262] In certain embodiments, the linker q is an integer greater than or equal to 0. In certain embodiments, q is an integer greater than or equal to 1.

[00263] In certain embodiments, for example, where the linker q is greater than 2, (A ^L ) _q is a group that is linked to the ULM, and A ^L ₁ and (A ^L ) _q are linked through linker building blocks (L) .

[00264] In certain embodiments, for example, where the linker q is 2, (A ^L ) _q is a group that is linked to A ^L ₁ and to the ULM.

[00265] In certain embodiments, for example, where the q of the linker is 1, the structure of the linker group L is -A ^L ₁ - and A ^L ₁ is a group that is linked to a ULM moiety and a PTM moiety.

[00266] In certain embodiments, the A ^L element of the linker (L) contains a group represented by a general structure selected from the group consisting of:

-NR(CH ₂ ) _n -(lower alkyl)-, -NR(CH ₂ ) _n -(lower alkoxy)-, -NR(CH ₂ ) _n -(lower alkoxy)-OSH ₂ -, -NR(CH ₂ ) _n -(lower alkoxy)-(lower alkyl)-OSH ₂ -, -NR(CH ₂ ) _n -(cycloalkyl)-(lower alkyl)-OSH ₂ -, -NR(CH ₂ ) _n -(heterocycloalkyl) -, -NR(CH ₂ CH ₂ O) _n -(lower alkyl)-O-CH ₂ -, -NR(CH ₂ CH ₂ O) _n -(heterocycloalkyl)-O-CH ₂ -, -NR(CH ₂ CH ₂ O) _n -aryl-O-CH ₂ -, -NR(CH ₂ CH ₂ O) _n -(heteroaryl)-O-CH ₂ -; -NR(CH ₂ CH ₂ O) _n -(cycloalkyl)-O-(heteroaryl)-O-CH ₂ -; -NR(CH ₂ CH ₂ O) _n -(cycloalkyl)-O-aryl-O-CH ₂ -, -NR(CH ₂ CH ₂ O) _n -(lower alkyl)-NH-aryl-O-CH ₂ -, -NR(CH ₂ CH ₂ O) _n -(lower alkyl)-O-aryl-CH ₂ , -NR(CH ₂ CH ₂ O) _n- cycloalkyl-O-aryl-, -NR(CH ₂ CH ₂ O) _n -cycloalkyl-O-(heteroaryl)1-, -NR(CH ₂ CH ₂ ) _n -(cycloalkyl)-O-(heterocyclyl)-CH ₂ , NR(CH ₂ CH ₂ ) _n -(heterocyclyl)- (heterocyclyl)-CH ₂ , -N(R1R2)-(heterocyclyl)-CH ₂ ; Where

n of the linker can be 0-10;

R of the linker may be H, lower alkyl;

R1 and R2 of the linker can form a ring with N attached.

[00267] In any aspect or embodiment described herein, the linker element A ^L (L) contains a group selected from:

Where:

*N of heterocycloalkyl shared with RTM; A

each m, n, o, p, q, r and s are independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 , 17, 18, 19 or 20.

[00268] In certain embodiments, the A ^L element of the linker (L) contains a group represented by a general structure selected from the group consisting of:

-N(R)-(CH2) _m -O(CH2) _n -O(CH2) _o -O(CH2) _p -O(CH2) _q -O(CH2) _r -OCH2-,

-O-(CH2) _m -O(CH2) _n -O(CH2) _o -O(CH2) _p -O(CH2) _q -O(CH2) _r -OCH2-,

-O-(CH2) _m -O(CH2) _n -O(CH2) _o -O(CH2) _p -O(CH2) _q -O(CH2) _r -O-;

-N(R)-(CH2) _m -O(CH2) _n -O(CH2) _o -O(CH2) _p -O(CH2) _q -O(CH2) _r -O-;

-(CH2) _m -O(CH2) _n -O(CH2) _o -O(CH2) _p -O(CH2) _q -O(CH2) _r -O-;

-(CH2) _m -O(CH2) _n -O(CH2) _o -O(CH2) _p -O(CH2) _q -O(CH2) _r -OCH2-;

m, n, o, p, q and r of the linker are independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 , 18, 19, 20;

when the quantity is zero, then there is no N-O or O-O bond

R of the linker is H, methyl and ethyl;

Linker X represents H and F

where m of the linker can be 2, 3, 4, 5

where each linker pit can independently be 0, 1, 2, 3, 4, 5, 6.

[00269] In any aspect or embodiment described herein, the linker element A ^L (L) is selected from the group consisting of:

where each type is independently selected from 0, 1, 2, 3, 4, 5 or 6.

[00270] In any aspect or embodiment described herein, the linker element A ^L is selected from the group consisting of:

where each m, n, o, p, q, r and s independently represents 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.

[00271] In any aspect or embodiment described herein, the linker element A ^L (L) is selected from the group consisting of:

[00272] In further embodiments, the linker (L) comprises a structure selected from, but not limited to, the structure shown below, wherein the dotted line denotes the point of attachment with PTM or ULM moieties:

Where:

W ^L1 and W ^L2 are each independently absent, a 4-8 membered ring with 0-4 heteroatoms, optionally substituted with R ^Q , wherein each R ^Q is independently H, halogen, OH, CN, CF ₃ , optionally substituted linear or branched C ₁ -C ₆ alkyl, optionally substituted linear or branched C ₁ -C ₆ alkoxy, or 2 R ^Q groups, taken together with the atom to which they are attached to form a 4-8 membered ring system containing 0-4 heteroatoms ;

Y ^L1 each independently represents a bond, optionally substituted linear or branched C ₁ -C ₆ alkyl and optionally one or more C atoms are replaced by O; or optionally substituted linear or branched C ₁ -C ₆ alkoxy;

n and m independently represent 0-10; And

denotes the point of attachment to PTM or ULM fragments.

[00273] In further embodiments, the linker (L) comprises a structure selected from, but not limited to, the structure shown below, wherein the dotted line denotes the point of attachment to the PTM or ULM moieties:

Where:

W ^L1 and W ^L2 are each independently absent, represent aryl, heteroaryl, cyclic group, heterocycle, C _1-6 alkyl and optionally one or more C atoms replaced by O or N, C _1-6 alkene and optionally one or more C atoms replaced by O, a C _1-6 alkyne and optionally one or more C atoms are replaced by O, a bicyclic group, biaryl, biheteroaryl or biheterocycle, each optionally substituted by R ^Q , each R ^Q independently represents H, halogen, OH, CN, CF ₃ , hydroxyl, nitrogen, C≡CH, C _2-6 alkenyl, C _2-6 alkynyl, optionally substituted linear or branched C ₁ -C ₆ alkyl, optionally substituted linear or branched C ₁ -C ₆ alkoxy, optionally substituted OC _1-3 alkyl (e.g. optionally substituted with 1 or more -F), OH, NH ₂ , NR ^Y1 R ^Y2 , CN or 2 R ^Q groups taken together with the atom to which they are attached to form a 4-8 membered system rings containing 0-4 heteroatoms; Y ^L1 each independently represents a bond, NR ^YL1 , O, S, NR ^YL2 , CR ^YL1 R ^YL2 , C=O, C=S, SO, SO ₂ , C ₁ -C ₆ alkyl (linear, branched, optionally substituted) and optionally one or more C atoms are replaced by O; optionally substituted linear or branched C ₁ -C ₆ alkoxy, 3-6 membered acyclic or aromatic ring with 0-4 heteroatoms;

^QL is a 3-6 membered acyclic or aromatic ring with 0-4 heteroatoms, optionally bridged, optionally substituted with 0-6 ^RQ , each ^RQ independently represents H, optionally substituted linear or branched _C1-6 alkyl (eg , optionally substituted with 1 or more halogens or C _1-6 alkoxyl), or 2 R ^Q groups, taken together with the atom to which they are attached to form a 3-8 membered ring system containing 0-2 heteroatoms;

R ^YL1 , R ^YL2 each independently represents H, OH, optionally substituted linear or branched C _1-6 alkyl (for example, optionally substituted 1 or more halogens or C _1-6 alkoxy), or R ¹ , R ² taken together with an atom , to which they join, forming a system of 3-8 membered rings containing 0-2 heteroatoms;

n and m independently represent 0-10; And

denotes the point of attachment to PTM or ULM fragments.

[00274] In further embodiments, the linker group is an optionally substituted (poly)ethylene glycol with from 1 to about 100 ethylene glycol units (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, etc., ethylene glycol elements), from about 1 to about 50 ethylene glycol elements, from 1 to about 25 ethylene glycol units, about 1 to about 10 ethylene glycol units, 1 to about 8 ethylene glycol units, 1 to 6 ethylene glycol units, 2 to 4 ethylene glycol units, or optionally substituted alkyl groups mixed with optionally substituted, O, N, S, P or Si atoms. In certain embodiments, the linker is substituted with aryl, phenyl, benzyl, alkyl, alkylene, or a heterocyclic group. In certain embodiments, the linker may be asymmetric or symmetric.

[00275] In any of the embodiments of the compounds described herein, the linker group can be any suitable moiety as described herein. In one embodiment, the linker is a substituted or unsubstituted polyethylene glycol group with a size ranging from about 1 to about 12 ethylene glycol units, from 1 to about 10 ethylene glycol units, from about 2 to about 6 ethylene glycol units, from about 2 to 5 ethylene glycol units, from about 2 to 4 elements of ethylene glycol.

[00276] In any aspect or embodiment described herein, the linker (L) includes an optionally substituted C ₁ -C ₅₀ alkyl (for example, C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , C ₇ , С ₈ , С ₉ , С ₁₀ , С ₁₁ , С ₁₂ , С ₁₃ , С ₁₄ , С ₁₅ , С 16, С ₁₇ , С ₁₈ , С ₁₉ , _{С 20 ,} С ₂₁ , С ₂₂ , С ₂₃ , С ₂₄ , С ₂₅ , С ₂₆ , С ₂₇ , С ₂₈ , С ₂₉ , С ₃₀ , С 31, С 32, С ₃₃ , С _{34 , С 35 ,} С ₃₆ , С ₃₇ , С ₃₈ , _{С 39 ,} С ₄₀ , C ₄₁ , C ₄₂ , C ₄₃ , C ₄₄ , C ₄₅ , C ₄₆ , C ₄₇ , C ₄₈ , C ₄₉ or C ₅₀ alkyl), where each carbon is optionally replaced by a heteroatom selected from N, S, P or Si atoms , which have the appropriate number of hydrogen atoms, substitutions, or both for full valency, provided that there is no heteroatom-heteroatom bond (e.g., the heteroatoms are not covalently bonded or located adjacent to each other).

[00277] In any aspect or embodiment described herein, the linker (L) includes from about 1 to about 50 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36 , 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50) alkylene glycol elements which are optionally substituted, wherein the carbon or oxygen may be replaced by a heteroatom selected from N atoms, S, P or Si with the appropriate number of hydrogen atoms for total valence. For example, in any aspect or embodiment described herein, the linker (L) has a chemical structure selected from:

, where carbon or oxygen may be replaced by a heteroatom selected from N, S, P or Si atoms with the appropriate number of hydrogen atoms for total valence, and m, n, o, p, q, r and s are independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20.

[00278] In another embodiment, the present invention is directed to a compound that contains a PTM group, as described above, that binds a target protein or polypeptide (e.g., Kirsten rat sarcoma protein (KRas or KRAS) and/or gain-of-function mutant KRas), which is ubiquitinated by a ubiquitin ligase, and is chemically linked directly or through a linker fragment L to a ULM group, or PTM is alternatively a ULM' group, which is also a ubiquitin ligase binding fragment, which may be the same as the ULM or different from the ULM group as described above, and is linked directly or via a linker fragment to the ULM group; a L is a linker moiety as described above, which may or may not be present, and which chemically (covalently) links ULM to PTM, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate or polymorph thereof.

[00279] In certain embodiments, the linker group L is a group containing one or more covalently linked structural elements independently selected from the group consisting of:

X is selected from the group consisting of O, N, S, S(O) and SO ₂ ; n is an integer from 1 to 5; R ^L1 represents hydrogen or alkyl, is a mono- or bicyclic aryl or heteroaryl, optionally substituted with 1-3 substituents selected from alkyl, halogen, haloalkyl, hydroxy, alkoxy or cyano; is a mono- or bicyclic cycloalkyl or heterocycloalkyl optionally substituted with 1-3 substituents selected from alkyl, halogen, haloalkyl, hydroxy, alkoxy or cyano; and the phenyl ring moiety may be optionally substituted with 1, 2 or 3 substituents selected from the group consisting of alkyl, halogen, haloalkyl, hydroxy, alkoxy and cyano. In an embodiment, the linker group L contains up to 10 covalently linked structural elements, as described above.

[00280] Although the ULM group and PTM group can be covalently linked to the linker group through any group that is suitable and stable for the chemical bond of the linker, in preferred aspects of the present invention, the linker is independently covalently linked to the ULM group and PTM group, preferably through an amide, ester, thioester, keto group, carbamate (urethane), carbon or ether, each of the groups can be included anywhere on the ULM group and PTM group, providing maximum binding of the ULM group to the ubiquitin ligase and the PTM group to the target protein for its degradation. (It should be noted that in some aspects, when the PTM group is a ULM group, the target protein for degradation may be the ubiquitin ligase itself). In certain preferred aspects, the linker may be linked to an optionally substituted alkyl, alkylene, alkene or alkyne group, aryl group or heterocyclic group on ULM and/or PTM groups.

Typical RTMs

[00281] In preferred aspects of the invention, the PTM group is a group that binds to target proteins. The targets of the PTM group are numerous in nature and are selected from proteins that are expressed in the cell such that at least a portion of the sequences are present in the cell and can bind to the PTM group. The term "protein" includes oligopeptides and polypeptide sequences of sufficient length so that they can bind to the PTM group in accordance with the present invention. Any protein in a eukaryotic system or microbial system, including a virus, bacteria or fungus, as otherwise described herein, is a target for ubiquitination mediated by the compounds of the present invention. Preferably, the target protein is a eukaryotic protein.

[00282] PTM groups according to the present invention include, for example, any fragment that specifically binds to a protein (binds to a target protein), and includes the following non-limiting examples of small molecule target protein fragments: KRas inhibitors, Hsp90 inhibitors, kinase inhibitors, HDM2 inhibitors & MDM2, compounds targeting human BET bromodomain containing proteins, HDAC inhibitors, human lysine methyltransferase inhibitors, angiogenesis inhibitors, nuclear hormone receptor compounds, immunosuppressive compounds and aryl hydrocarbon receptor (AHR) targeting compounds and many others. The compositions described below are examples of some members of small molecule fragments that bind the target protein. Such target protein binding fragments of small molecules also include pharmaceutically acceptable salts, enantiomers, solvates and polymorphs of these compositions, as well as other small molecules that can target the protein of interest, such as KRas and/or mutant KRas, including mutant(s) ) KRas with function enhancement such as KRas ^G12C . These binding fragments are linked to a ubiquitin ligase binding fragment, preferably through a linker to present the target protein (to which the target protein fragment is bound), such as KRas and/or gain-of-function mutant(s) of KRas, in close proximity to the ubiquitin ligase for ubiquitination and degradation.

[00283] The present invention can be used to treat a variety of disease states and/or pathological conditions, including any disease state and/or pathological condition in which proteins are dysregulated and where the patient would benefit from protein degradation and/or inhibition.

[00284] In a further aspect, the disclosure provides therapeutic compositions containing an effective amount of a compound as described herein, or a salt form thereof, and a pharmaceutically acceptable carrier, additive or excipient and, optionally, an additional bioactive agent. Therapeutic compositions modulate protein degradation in a patient or subject, for example, an animal such as a human, and can be used to treat or improve disease states or conditions that are modulated by the degraded protein. In certain embodiments, the therapeutic compositions described herein can be used to cause the degradation of proteins of interest to treat or alleviate a disease, such as cancer (including, for example, pancreatic cancer, colon cancer, lung cancer, non-small cell lung cancer, or their combination). In certain further embodiments, the disease includes or is pancreatic cancer, colon cancer, colorectal cancer, lung cancer, non-small cell lung cancer, biliary tract malignancies, endometrial cancer, cervical cancer, bladder cancer, liver cancer, myeloid leukemia, breast cancer or a combination of both.

[00285] In alternative aspects, the present invention provides a method of treating a disease state or alleviating the symptoms of a disease or condition in a subject in need thereof by degrading a protein or polypeptide by which the disease state or condition is modulated, comprising administering to said patient or subject an effective an amount, for example, a therapeutically effective amount of at least one compound as described above, optionally in combination with a pharmaceutically acceptable carrier, additive or excipient, and optionally with an additional bioactive agent, wherein the composition is effective for treating or alleviating a disease or disorder or their symptom in the subject. The method according to the present invention can be used to treat a large number of disease states or pathological conditions, including cancer (including, for example, pancreatic cancer, colon cancer, colorectal cancer, lung cancer, non-small cell lung cancer, biliary tract malignancies, endometrial cancer, cancer cervical cancer, bladder cancer, liver cancer, myeloid leukemia, breast cancer, or a combination thereof) resulting from the administration of effective amounts of at least one compound described herein. The disease state or pathological condition may be a disease caused by a microbial agent or other exogenous agent, such as a virus, bacteria, fungus, protozoa or other microbe, or may be a disease state that is caused by overexpression of a protein, resulting in a disease state and/or pathological condition.

[00286] In another aspect, the description provides methods for identifying the effects of degradation of proteins of interest in a biological system using compounds of the present invention.

[00287] The term “target protein” is used herein and below to describe a protein or polypeptide that is targeted for binding to a compound of the invention and degradation by a ubiquitin ligase. For example, in any aspect or embodiment described herein, the PTM is a small molecule containing a KRas protein targeting moiety. Such small molecule fragments that bind the target protein also include pharmaceutically acceptable salts, enantiomers, solvates and polymorphs of these compositions, as well as other small molecules that can target the protein of interest. These linking moieties are linked to at least one ULM group (e.g., VLM, CLM, ILM, and/or MLM) via at least one L linker group.

[00288] Target proteins that can be bound to a target protein fragment and degradable by a ligase to which the ubiquitin ligase binding fragment is bound include any protein or peptide, including fragments thereof, analogs thereof and/or homologs thereof. Target proteins include proteins and peptides having any biological function or activity, including structural, regulatory, hormonal, enzymatic, genetic, immunological, contractile, storage, transport and signal transduction. For example, in any aspect or embodiment described herein, the PTM is a KRas protein binding fragment.

[00289] These various target proteins, such as the KRas protein, can be used in screens that identify fragments of a compound that bind to the protein, and by incorporating the fragment into compounds of the invention, the level of activity of the protein can be altered for therapeutic purposes as the final result.

[00290] The term "target protein binding fragment" or PTM is used to describe a small molecule that binds to a target protein or other protein or polypeptide of interest and places/presents that protein or polypeptide in close proximity to a ubiquitin ligase, resulting in possible degradation protein or polypeptide ubiquitin ligase. Non-limiting examples of small molecule target protein binding fragments include KRas inhibitors, Hsp90 inhibitors, kinase inhibitors, MDM2 inhibitors, compounds targeting human BET bromodomain containing proteins, HDAC inhibitors, human lysine methyltransferase inhibitors, angiogenesis inhibitors, immunosuppressive compounds and compounds targeting aryl hydrocarbon receptor (AHR) and many others. The compositions described below are examples of some of the small molecule members of the target proteins. Exemplary target protein fragments of the present invention include KRas inhibitors, haloalkanghalogenase inhibitors, Hsp90 inhibitors, kinase inhibitors, MDM2 inhibitors, compounds targeting human BET bromodomain containing proteins, HDAC inhibitors, human lysine methyltransferase inhibitors, angiogenesis inhibitors, immunosuppressive compounds and compounds targeting aryl hydrocarbon receptor (AHR).

[00291] The compositions described herein are examples of some members of these types of small molecule fragments that bind a target protein. Such small molecule fragments that bind the target protein also include pharmaceutically acceptable salts, enantiomers, solvates and polymorphs of these compositions, as well as other small molecules that can target the protein of interest.

[00292] In any aspect or embodiment described herein, the PTM is a KRas protein binding/targeting fragment, for example, a small molecule containing a KRas protein binding/targeting fragment. In any aspect or embodiment described herein, PTM binds a mutant KRas, such as a gain-of-function mutant of KRas (such as KRas ^G12C ). In any aspect or embodiment described herein, PTM has a chemical structure represented by:

Where:

is aryl, heteroaryl, cycloalkyl or heterocycloalkyl; X _PTM is C or N;

WPTM is selected from the group consisting of optionally substituted C ₃ -C ₆ cycloalkyl and optionally substituted C ₃ -C ₆ heteroalkyl, optionally substituted C ₃ -C ₆ heterocycloalkyl, optionally substituted aryl (for example, optionally substituted C ₅ -C ₇ aryl), optionally substituted heteroaryl (eg, optionally substituted C ₅ -C ₇ heteroaryl);

R _PTM1A is NR _PTM9 R _PTM10 , OR _PTM9 R _PTM10 , H, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted C3-C6 cycloalkyl, optionally substituted O-(C3-C6 cycloalkyl), optionally substituted C3-C6 heteroalkyl, optional substituted -O-C _1-4 alkyl-C _3-6 cycloalkyl, optionally substituted O-(C3-C6 heteroalkyl), optionally substituted OC _1-4 alkyl-C _3-6 heteroalkyl, optionally substituted O-C _1-4 alkyl -C _3-6 heterocycloalkyl, optionally substituted aryl (e.g., optionally substituted C5-C7 aryl), optionally substituted O-aryl (e.g., optionally substituted O-(C5-C7 aryl)), optionally substituted heteroaryl (e.g., optionally substituted C5 -C7 heteroaryl), optionally substituted O-heteroaryl (e.g., optionally substituted O-(C5-C7 heteroaryl)), optionally substituted (for example, optionally substituted with at least one alkyl, such as * carbon, which may be optionally substituted with alkyl), optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted (e.g., optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), optionally substituted (e.g., optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), optionally substituted (e.g., optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), optionally substituted (e.g., optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), optionally substituted (e.g., optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), optionally substituted (eg, optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), where N* represents the N atom of a heterocycloalkyl (eg, C4-C8 heterocycloalkyl) of the linker (L); RP _TM1B is NR _PTM9 R _PTM10 , OR _PTM9 R _PTM10 , H, optionally substituted alkyl, optionally substituted O-alkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted O-(C3-C6 cycloalkyl), optionally substituted -O-C _1-4 alkyl-C _3-6 cycloalkyl, optionally substituted C3-C6 heteroalkyl, optionally substituted O-(C3-C6 heteroalkyl), optionally substituted OC _1-4 alkyl-C _3-6 heteroalkyl, optionally substituted aryl (e.g., optionally substituted C5-C7 aryl), optionally substituted O-aryl (e.g., optionally substituted O-(C5-C7 aryl)), optionally substituted heteroaryl (e.g., optionally substituted C5-C7 heteroaryl), optionally substituted O-heteroaryl (e.g., optionally substituted O(C5-C7 heteroaryl)), optionally substituted (e.g., optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted

R _PTM9 and R _PTM10 are each independently H, optionally substituted C ₁ -C ₆ alkyl, optionally substituted aliphatic amine, optionally substituted aliphatic amide;

R _PTM2 is H, (C=O) R _PTM2 '; optionally substituted straight or branched alkyl;

R _PTM2 ' is an optionally substituted straight or branched alkyl, an optionally substituted alkene, -N(R _PTM8 ) ₂ or -C(OH) ₂ ;

R _PTM3 is alkyl, alkoxy, phenyl, or naphthalene, each independently substituted with OH, H, halogen;

R _PTM4A represents OH, H, halogen, optionally substituted linear or branched C ₁ -C ₆ alkyl;

R _PTM4B represents OH, H, halogen, optionally substituted linear or branched C ₁ -C ₆ alkyl;

R _PTM5 is selected from the group consisting of optionally substituted aryl, optionally substituted biaryl, optionally substituted heteroaryl, optionally substituted biheteroaryl, optionally substituted C ₃ -C ₆ cycloalkyl, optionally substituted C ₃ -C ₆ cycloheteroalkyl, halogen, H, optionally substituted linear or branched alkyl (eg, optionally substituted linear or branched C ₁ -C ₆ alkyl), OH and alkoxy;

R _PTM8 is H or alkyl (eg, C ₁ alkyl, C ₂ alkyl, C ₃ alkyl or C ₄ alkyl);

t represents 0, 1, 2, 3, 4, 5, 6 (such as 0, 1, 2, 3); And

denotes the site of attachment of at least one of the linkers, ULM, ULM', CLM, CLM', VLM, VLM', ILM, ILM', MLM, MLM', or combinations thereof.

[00293] In any aspect or embodiment described herein, PTM has a chemical structure represented by:

Where:

X _PTM1 is NH or O;

R _PTM6 is aryl, heteroaryl, where N* represents the N atom of a heterocycloalkyl (eg C4-C8 heterocycloalkyl) linker (L);

R _PTM7 is H, aryl, O-aryl, heteroaryl, O-heteroaryl,

t represents 0, 1, 2, 3, 4, 5, 6 (for example, 1, 2 or 3);

R _PTM9 is H, an optionally substituted C1-C6 alkyl, an optionally substituted aliphatic amine, an optionally substituted aliphatic amide, or an optionally substituted (eg, optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl);

may be a single bond or a double bond; And

denotes the site of attachment of at least one of the linker, ULM, ULM', CLM, CLM', VLM, VLM', ILM, ILM', MLM, MLM', or a combination thereof.

[00294] In any aspect or embodiment described herein, the PTM is selected from:

[00295] In any aspect or embodiment described herein, the PTM is: (i) a PTM selected from the combination of tables 4, 6, 8, 10 and 12; or (ii) RTM Table 1.

Therapeutic compositions

[00296] Pharmaceutical compositions containing combinations of an effective amount of at least one bifunctional compound as described herein and one or more compounds otherwise described herein, all in effective amounts in combination with a pharmaceutically effective amount of a carrier, additive, or filler, represent an additional aspect of the present invention.

[00297] The present invention includes, where applicable, compositions containing pharmaceutically acceptable salts, in particular acid or base addition salts of compounds as described herein. The acids that are used to prepare pharmaceutically acceptable acid addition salts of the above basic compounds useful in this aspect are acids that form non-toxic acid addition salts, i.e. salts containing pharmacologically acceptable anions such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, acid citrate, tartrate, bitartrate, succinate, maleate, fumarate, gluconate, saccharate, benzoate , methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate salts [ie, 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)] and many others.

[00298] Pharmaceutically acceptable base addition salts can also be used to prepare pharmaceutically acceptable salt forms of the compounds or derivatives of the present invention. Chemical bases that can be used as reagents to prepare pharmaceutically acceptable base salts of the present compounds that are acidic in nature are those that form non-toxic base salts with such compounds. Such non-toxic base salts include, but are not limited to, salts derived from pharmacologically acceptable cations such as alkali metal cations (eg, potassium and sodium) and alkaline earth metal cations (eg, calcium, zinc and magnesium), ammonium or water-soluble addition salts amines such as N-methylglucamine (meglumine), and lower alkanolammonium salts and other bases of pharmaceutically acceptable organic amines, among others.

[00299] The compounds as described herein can, in accordance with the invention, be administered in single or divided doses by the oral, parenteral or topical routes. Administration of the active compound may range from continuous (intravenous drip) to multiple oral administrations per day (eg, QID) and may include oral, topical, parenteral, intramuscular, intravenous, subcutaneous, transdermal (which may include enhanced penetration of the agent), buccal , sublingual and suppository administration, among other routes of administration. Enteric-coated oral tablets can also be used to increase the bioavailability of compounds via the oral route of administration. The most effective dosage form will depend on the pharmacokinetics of the particular agent chosen, as well as the severity of the patient's disease. It is also possible to administer the compounds of the invention in the form of sprays, mist aerosols or aerosols for intranasal, intratracheal or pulmonary administration. The present invention, therefore, also relates to pharmaceutical compositions containing an effective amount of a compound as described herein, optionally in combination with a pharmaceutically acceptable carrier, additive or excipient. The compounds of the invention may be administered in immediate release, intermediate release, or sustained or controlled release forms. Sustained or controlled release forms are preferably administered orally, but also as suppositories, transdermal or other topical forms. Intramuscular injections in liposomal form can also be used to control or maintain release of the compound at the injection site.

[00300] The compositions described herein can be formulated in conventional manner using one or more pharmaceutically acceptable carriers, and can also be administered in controlled release formulations. Pharmaceutically acceptable carriers that may be used in these pharmaceutical compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffers such as phosphates, glycine, sorbic acid , potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as prolamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol , sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol and lanolin.

[00301] The compositions described herein can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or through an implanted reservoir. As used herein, the term “parenteral” includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion routes. Preferably, the compositions are administered orally, intraperitoneally or intravenously.

[00302] Sterile injectable forms of compositions as described herein may be aqueous or oleaginous suspensions. These suspensions can be formulated in accordance with techniques known in the art, using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, a solution in 1,3-butanediol. Acceptable vehicles and solvents that may be used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils are commonly used as a solvent or suspending medium. Any soft, fixed oil can be used for this purpose, including synthetic mono- or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives are useful in the preparation of injectable preparations, as are natural pharmaceutically acceptable oils such as olive oil or castor oil, especially in their polyoxyethylene versions. These oil solutions or suspensions may also contain a long chain alcohol diluent or dispersant such as Ph. Helv or similar alcohol.

[00303] The pharmaceutical compositions described herein can be administered orally in any orally acceptable dosage form, including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of oral tablets, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate are also commonly added. For oral administration in capsule form, suitable diluents contain lactose and dried corn starch. When aqueous suspensions are required for oral administration, the active ingredient is combined with emulsifying and suspending agents. Certain sweeteners, flavors or colors may also be added if necessary.

[00304] Alternatively, the pharmaceutical compositions described herein can be used in the form of suppositories for rectal administration. They can be prepared by mixing the agent with a suitable non-irritant excipient that is solid at room temperature but liquid at rectal temperature and will therefore melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

[00305] The pharmaceutical compositions described herein can also be applied topically. Suitable topical formulations are readily available for each of these areas or organs. Topical administration to the lower intestinal tract can be in the form of rectal suppositories (see above) or suitable enema formulations. Transdermal patches that are acceptable for topical use may also be used.

[00306] For topical use, the pharmaceutical compositions may be formulated as a suitable ointment containing the active ingredient suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of the present invention include, but are not limited to, mineral oil, liquid petrolatum, medicinal petrolatum, propylene glycol, polyoxyethylene, a polyoxypropylene compound, emulsifying wax and water. In some preferred aspects of the invention, the compounds can be applied to a stent that is to be surgically implanted in a patient to suppress or reduce the likelihood of occlusion occurring in the stent in the patient.

[00307] Alternatively, the pharmaceutical compositions may be formulated as a suitable lotion or cream containing the active ingredients suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

[00308] For ophthalmic use, pharmaceutical compositions can be formulated as micronized suspensions in isotonic, sterile, pH-adjusted saline, or preferably as solutions in isotonic, sterile, pH-adjusted saline, with or without the addition of a preservative such as benzalkonium chloride. Alternatively, for ophthalmic use, the pharmaceutical compositions may be formulated as an ointment such as petrolatum.

[00309] The pharmaceutical compositions described herein can also be administered as a nasal aerosol or inhalation. Such compositions are prepared according to methods well known in the pharmaceutical art and can be formulated as solutions in saline using benzyl alcohol or other suitable preservatives, absorption catalysts to enhance bioavailability, fluorocarbons and/or other conventional solubilizing or dispersing agents. .

[00310] The amount of compound in the pharmaceutical composition described herein that can be combined with carrier materials to form a unit dosage form will vary depending on the host and disease being treated and the particular route of administration. Preferably, the compositions will be formulated to contain from about 0.05 milligrams to about 750 milligrams or more, more preferably from about 1 milligram to about 600 milligrams, and even more preferably from about 10 milligrams to about 500 milligrams of the active ingredient, alone or in combination with at least one of the other compounds according to the invention.

[00311] It should also be understood that the specific dosage and treatment regimen for any particular patient will depend on numerous factors, including the potency of the particular compound used, age, body weight, general health, gender, diet, time of administration, rate of elimination, drug combination , the opinion of the attending physician and the severity of the specific disease or condition being treated.

[00312] A patient or subject in need of therapy using compounds in accordance with the methods described herein can be treated by administering to the patient (subject) an effective amount of a compound according to the invention, including pharmaceutically acceptable salts, solvates or polymorphs thereof, optionally in a pharmaceutically acceptable carrier or diluent, either alone or in combination with other known therapeutic agents as otherwise specified herein.

[00313] These compounds can be administered by any suitable route, such as orally, parenterally, intravenously, intradermally, subcutaneously or topically, including transdermally, in liquid, cream, gel or solid or aerosol form.

[00314] The active compound is included in a pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver to a patient a therapeutically effective amount for the desired indication without causing serious toxic effects in the patient being treated. The preferred dose of the active compound for all of the above conditions is in the range of from about 10 ng/kg to 300 mg/kg, preferably from 0.1 to 100 mg/kg per day, more often from about 0.5 to 25 mg per kilogram of body weight of the recipient /patient per day. Typical topical dosage will be in the range of 0.01-5% w/w. in a suitable medium.

[00315] The compound is conveniently administered in any suitable unit dosage form, including, but not limited to, a form containing less than 1 mg, 1 to 3000 mg, preferably 5 to 500 mg of active ingredient per unit dosage form. An oral dose of approximately 25-250 mg is often convenient.

[00316] The active ingredient is preferably administered to achieve peak plasma concentrations of the active compound of about 0.00001-30 mM, preferably about 0.1-30 μM. This can be achieved, for example, by intravenous injection of a solution or formulation of the active ingredient, optionally in saline or aqueous media, or by administering a bolus of the active ingredient. Oral administration is also suitable for achieving effective plasma concentrations of the active agent.

[00317] The concentration of the active compound in the drug composition will depend on the rate of absorption, distribution, inactivation and elimination of the drug, as well as other factors known to those skilled in the art. It should be noted that dosage values will also vary depending on the severity of the condition that needs to be alleviated. It should also be understood that for any particular subject, specific dosage regimens should be adjusted over time according to individual needs and the professional judgment of the person prescribing or supervising the administration of the compositions, and that the concentration ranges set forth herein are only approximate and should not be used be considered as limiting the scope or practice of use of the claimed composition. The active ingredient may be administered as a single dose or may be divided into several smaller doses to be administered at varying intervals.

[00318] Oral compositions typically contain an inert diluent or edible carrier. They may be enclosed in gelatin capsules or compressed into tablets. For oral therapeutic administration, the active compound or a prodrug derivative thereof may be included in excipients and used in the form of tablets, lozenges or capsules. Pharmaceutically compatible binding agents and/or adjuvant materials may be included as part of the composition.

[00319] Tablets, pills, capsules, lozenges, etc. may contain any of the following ingredients or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a dispersing agent such as alginic acid, Primogel or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring such as peppermint, methyl salicylate, or orange flavor. When the unit dosage form is a capsule, it may contain, in addition to material of the above type, a liquid carrier such as a fatty oil. In addition, unit dosage forms may contain various other materials that modify the physical form of the dosage unit, such as coatings of sugar, shellac, or enteric agents.

[00320] The active compound or a pharmaceutically acceptable salt thereof can be used as a component of an elixir, suspension, syrup, wafer, chewing gum, or the like. The syrup may contain, in addition to the active compounds, sucrose as a sweetening agent, as well as some preservatives, dyes, coloring agents and flavorings.

[00321] The active compound or pharmaceutically acceptable salts thereof may also be mixed with other active materials that do not impair the desired effect, or with materials that complement the desired effect, such as anticancer agents, including epidermal growth factor receptor inhibitors, EPO (Erythropoietin, erythropoietin ) and darbapoetin alfa, among others. In some preferred aspects of the invention, one or more compounds of the invention are co-administered with another biologically active agent, such as an anticancer agent or an antibiotic-containing healing agent, as otherwise described herein.

[00322] Solutions or suspensions used for parenteral, intradermal, subcutaneous or topical use may include the following components: a sterile diluent such as water for injection, saline, fatty oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methylparabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates; and tone-regulating agents such as sodium chloride or dextrose. The parent drug may be contained in ampoules, disposable syringes, or multiple dose vials made of glass or plastic.

[00323] For intravenous administration, the preferred carriers are saline or phosphate buffered saline (PBS).

[00324] In one embodiment, the active compounds are formulated with carriers that will protect the compound from rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters and polylactic acid can be used. Methods for preparing such compositions will be known to those skilled in the art.

[00325] Pharmaceutically acceptable carriers may also be liposomal suspensions. They can be prepared by methods known to those skilled in the art, for example, as described in US Pat. No. 4,522,811 (which is incorporated herein by reference in its entirety). For example, liposomal preparations can be prepared by dissolving the appropriate lipid(s) (such as stearoylphosphatidylethanolamine, stearoylphosphatidylcholine, aracadoylphosphatidylcholine and cholesterol) in an inorganic solvent, which is then evaporated, leaving behind a thin film of dried lipid on the surface of the container. An aqueous solution of the active compound is then introduced into the container. The container is then manually rotated to release lipid material from the side walls of the container and disperse the lipid aggregates, thereby forming a liposome suspension.

Therapeutic methods

[00326] In a further aspect, the disclosure provides therapeutic compositions containing an effective amount of a compound as described herein, or a salt form thereof, and a pharmaceutically acceptable carrier. Therapeutic compositions modulate protein degradation in a patient or subject, for example an animal such as a human, and can be used to treat or alleviate disease states or pathological conditions that are modulated by the degraded protein.

[00327] The terms “treat,” “cure,” and “therapy,” etc., as used herein, refer to any action beneficial to a patient to whom the compounds of the invention may be administered, including the treatment of any pathological or disease state modulated via a protein to which the proposed compounds bind. Diseaseful or pathological conditions, including cancer (eg, at least one of the following: pancreatic cancer, colon cancer, colorectal cancer, lung cancer, non-small cell lung cancer, biliary tract malignancies, endometrial cancer, cervical cancer, bladder cancer , liver cancer, myeloid leukemia, breast cancer or combinations thereof) that can be treated using the compounds of the invention are set out above.

[00328] The invention provides therapeutic compounds provided herein for degrading target proteins to treat or alleviate a disease such as cancer (e.g., pancreatic cancer, colon cancer, colorectal cancer, lung cancer, non-small cell lung cancer, biliary tract malignancies, endometrial cancer, cervical cancer, bladder cancer, liver cancer, myeloid leukemia or breast cancer). In some further embodiments, the disease is multiple myeloma. As such, in another aspect, the present invention provides a method for ubiquitination/degradation of a target protein in a cell. In some embodiments, the method comprises administering a bifunctional compound, as described herein, containing, for example, a ULM and a PTM, preferably linked through a linker moiety, as otherwise described herein, wherein the ULM is linked to the PTM, and wherein the ULM recognizes a ubiquitin pathway protein (e.g., a ubiquitin ligase, such as E3 ubiquitin ligase, including cereblon, VHL, IAP, and/or MDM2), and the PTM recognizes the target protein such that when the target protein is placed in close proximity to the ubiquitin ligase, degradation of the target protein occurs, resulting in degradation/inhibition action of the target protein and control of protein levels. The control of protein levels provided by the present invention provides treatment for a disease state or pathological condition that is modulated by a target protein by reducing the level of that protein in cells, eg, patient cells. In some embodiments, the method includes administering an effective amount of a compound as described herein, optionally containing a pharmaceutically acceptable excipient, carrier, adjuvant, other bioactive agent, or a combination thereof.

[00329] In additional embodiments, the disclosure provides methods for treating or alleviating a disease, disorder, or symptom thereof in a subject or patient, e.g., an animal such as a human, comprising administering to the subject in need thereof a composition containing an effective amount, e.g., therapeutically an effective amount of a compound as described herein, or a salt form thereof, and a pharmaceutically acceptable excipient, carrier, adjuvant, other biologically active agent, or a combination thereof, wherein the composition is effective for treating or alleviating a disease or disorder or symptom thereof in a subject.

[00330] In another aspect, the description provides methods for identifying the effects of degradation of proteins of interest in a biological system using compounds of the invention.

[00331] In another embodiment, the present invention is directed to a method of treating a human patient in need of modulating a disease or pathological condition with a protein, wherein degradation of the protein will produce a therapeutic effect in the patient, the method comprising administering to the patient in need thereof, an effective amount of a compound of the invention, optionally in combination with another bioactive agent. The disease or pathological condition may be a disease caused by a microbial agent or other exogenous agent, such as a virus, bacteria, fungus, protozoa or other microbe, or may be a disease condition that is caused by overexpression of a protein, resulting in a disease and/or pathological condition.

[00332] The term "disease or pathological condition" is used to describe any disease or pathological condition in which dysregulation of a protein occurs (i.e., the amount of protein expressed in the patient is increased) and where destruction of one or more proteins in the patient can provide beneficial therapy or symptom relief for a patient in need. In some cases, a painful or pathological condition can be cured.

[00333] Disease or pathological conditions that can be treated using the compounds of the present invention include, for example, asthma, autoimmune diseases such as multiple sclerosis, various types of cancer, ciliopathies, cleft palate, diabetes, heart disease, hypertension, inflammatory disease intestinal, mental retardation, mood disorder, obesity, refractive error, infertility, Angelman syndrome, Canavan disease, celiac disease, Charcot-Marie-Tooth disease, cystic fibrosis, Duchenne muscular dystrophy, hemochromatosis, hemophilia, Klinefelter syndrome, neurofibromatosis, fibromatosis, fibromatous disease , (PKD1) or 4 (PKD2) Prader-Willi syndrome, sickle cell anemia, Tay-Sachs disease, Turner syndrome.

[00334] The term "neoplasia" or "cancer" is used throughout this specification to refer to a pathological process that results in the formation and growth of a cancerous or malignant neoplasm, that is, abnormal tissue that grows by cell proliferation, often more rapidly than normal, and continues to grow after the cessation of the incentives that initiated the cessation of growth. In any aspect or embodiment described herein, the disease or disorder is cancer or neoplasia selected from pancreatic cancer, colon cancer, colorectal cancer, lung cancer, non-small cell lung cancer, biliary tract malignancies, endometrial cancer, cancer cervical cancer, bladder cancer, liver cancer, myeloid leukemia, or breast cancer (eg, cancer or neoplasia selected from pancreatic cancer, colon cancer, lung cancer, or non-small cell lung cancer). Malignant neoplasms exhibit partial or complete lack of structural organization and functional coordination with normal tissue, and most of them invade surrounding tissue, metastasize to multiple sites and are likely to recur after attempted removal, and cause death in the patient if not properly treated. As used herein, the term “neoplasia” is used to describe all malignant disease states and includes or covers the pathological process associated with malignant hematogenous, ascitic and solid tumors. Exemplary cancers that may be treated with the present compounds, either alone or in combination with at least one additional anticancer agent, include squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, hepatocellular carcinoma and renal cell carcinoma, bladder, colon, breast glands, cervix, colon, esophagus, head, kidney, liver, lungs, neck, ovaries, pancreas, prostate and stomach; leukemia; benign and malignant lymphomas, especially Burkitt's lymphoma and non-Hodgkin's lymphoma; benign and malignant melanomas; myeloproliferative diseases; sarcomas, including Ewing's sarcoma, hemangiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcoma, peripheral neuroepithelioma, synovial sarcoma, gliomas, astrocytomas, oligodendrogliomas, ependymomas, glioblastomas, neuroblastomas, ganglioneuromas, gangliogliomas, medulloblastomas , pineal cell tumors, meningiomas, meningeal sarcomas, neurofibromas, neuromas; colon cancer, breast cancer, prostate cancer, non-small cell lung cancer, biliary tract malignancies, endometrial cancer, myeloid leukemia, cervical cancer, uterine cancer, lung cancer, ovarian cancer, testicular cancer, thyroid cancer, astrocytoma, esophageal cancer, pancreatic cancer, stomach cancer, liver cancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's disease, Wilms tumor and teratocarcinomas. Additional types of cancer that can be treated using the compounds of the present invention include, for example, T-lineage acute lymphoblastic leukemia (T-ALL), T-lineage lymphoblastic lymphoma (T-LL), peripheral T-cell lymphoma, T-cell adult leukemia, pre-B cell acute lymphoblastic leukemia (ALL, acute lymphoblastic leukemia), pre-B cell lymphomas, large B cell lymphoma, Burkits lymphoma, B cell ALL, Philadelphia chromosome ALL, and Philadelphia chromosome chronic myelocytic leukemia chromosome (CML, Chronic myeloid leukaemia).

[00335] The term "bioactive agent" is used to describe an agent, other than a compound of the invention, that is used in combination with the compounds of the invention as an agent with biological activity to assist in the intended therapy, inhibition and/or prevention/prophylaxis, for which the proposed connections are used. Preferred bioactive agents for use herein include agents that have pharmacological activity similar to that for which the present compounds are used or administered and include, for example, anticancer agents, antiviral agents, especially, but not limited to, anti-HIV agents and anti-viral agents. HCV (Hepatitis C Virus, hepatitis C virus), antimicrobial agents, antifungal agents, etc.

[00336] The term "additional anticancer agent" is used to describe an anticancer agent that can be combined with the compounds of the invention to treat cancer. These agents include, for example, everolimus, trabectin, abraxane, TLK 286, AV-299, DN-101, pazopanib, GSK690693, RTA 744, ON 0910.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258 , GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263, FLT-3 inhibitor, VEGFR inhibitor, EGFR TK inhibitor, Aurora kinase inhibitor, modulator PIK-1, Bcl-2 inhibitor, HDAC inhibitor, c-MET inhibitor, PARP inhibitor, Cdk inhibitor, EGFR TK inhibitor, IGFR-TK inhibitor, anti-HGF antibody, PI3 kinase inhibitor, AKT inhibitor, mTORC1/2 inhibitor, JAK inhibitor /STAT, immune checkpoint inhibitor 1 or 2, focal adhesion kinase inhibitor, Map mitogen-activated protein kinase inhibitor (mek), VEGF antibody drug, pemetrexed, erlotinib, dasatanib, nilotinib, decatanib, panitumumab, amrubicin, oregovomab, liposome-encapsulated paclitaxel , easy to use, nolatrexed, azd2171, batabulin, ofatumumab, zanolimumab, edotecarin, tetrandrine, rubitecan, tesimmilifen, oblimersen, ticilimumab, ipilimumab, gossypol, Bio 111, 131-I-TM-601, ALT-110, BIO 140, CC 8490, cilengitide, gimatecan, IL13-PE38QQR, INO 1001, IPdRl KRX-0402, lucanton, LY317615, neuradiab, vitespan, Rta 744, Sdx 102, talampanel, atrasentan, Xr 311, romidepsin, ADS-100380 , sunitinib, 5-fluorouracil , vorinostat, etoposide, gemcitabine, doxorubicin, liposomal doxorubicin, 5'-deoxy-5-fluorouridine, vincristine, temozolomide, ZK-304709, seliciclib; PD0325901, AZD-6244, capecitabine, L-glutamic acid, N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H-pyrrolo[2,3-d]pyrimidine-5- yl)ethyl]benzoyl]-, disodium salt, heptahydrate, camptothecin, PEG-labeled irinotecan, tamoxifen, toremifene citrate, anastrazole, exemestane, letrozole, DES (diethylstilbestrol), estradiol, estrogen, conjugated estrogen, bevacizumab, IMC-1C11, CHIR- 258); 3-[5-(methylsulfonylpiperadinemethyl)-indolylquinolone, vatalanib, AG-013736, AVE-0005, goserelin acetate, leuprolide acetate, triptorelin pamoate, medroxyprogesterone acetate, hydroxyprogesterone caproate, megestrol acetate, raloxifene, bicalutamide, flute amide, nilutamide, megestrol acetate, SR-724714; TAK-165, HKI-272, erlotinib, lapatanib, canertinib, anti-ABX-EGF, erbitux, EKB-569, PKI-166, GW-572016, ionafarnib, BMS-214662, tipifarnib; amifostine, NVP-LAQ824, suberoylanalide hydroxamic acid, valproic acid, trichostatin A, FK-228, SU11248, sorafenib, KRN951, aminoglutethimide, arsacrine, anagrelide, L-asparaginase, Bacillus Calmette-Guerin B vaccine CG), adriamycin , bleomycin, buserelin, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, clodronate, cinroterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, diethylstilbestrol, epirubicin, fludarabine, fludrocortisone, fluoxymesterone, flutamide, Gleevec, gemcitabine , hydroxyurea, idarubicin, ifosfamide , imatinib, leuprolide, levamisole, lomustine, mechlorethamine, melphalan, 6-mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, octreotide, oxalplatin, pamidronate, pentostatin, plicamycin, porfimer, procarbazine, raltitrexed, rituk Simab, streptozocin, teniposide , testosterone, thalidomide, thioguanine, thiotepa, tretinoin, vindesine, 13-cis-retinoic acid, phenylalanine mustine, uramustine, estramustine, altretamine, floxuridine, 5-deooxyuridine, cytosine arabinoside, 6-mecaptopurine, deoxycoformycin, calcitriol, valrubicin, mi Tramycin, Vinblastine, vinorelbine, topotecan, razoxin, marimastat, COL-3, neovastat, BMS-275291, squalamine, endostatin, SU5416, SU6668, EMD121974, interleukin-12, IM862, angiostatin, Vitaxin, droloxifene, idoxifene, spironolactone, finasteride, cimitidine, trastuzuma b, Denileikin Diftitox, Gephitinib, Bortesimib, Paklitaksel, Paklitaksel without crephor, Docatoxel, Epitilo B, BMS-247550, BMS-310705, Drooloxifen, 4-Hydroxitamoxifen, Pipenxifen, Era-923, Arzoxifen, Fulvestrant, Acorus, Acorus, Acorus. , Lazophoxifen, Idoksifen, TSE- 424, HMR-3339, ZK186619, topotecan, PTK787/ZK 222584, VX-745, PD 184352, rapamycin, 40-O-(2-hydroxyethyl) rapamycin, temsirolimus, AR-23573, RAD001, AVT-578, VS-210 , LY294002, LY292223, LY292696, LY293684, LY293646, wortmannin, ZM336372, L-779,450, PEG-filgrastim, darbepoetin, erythropoietin, granulocyte colony-stimulating factor, zoledronate, prednisone, cetuxime ab, granulocyte-macrophage colony-stimulating factor, histrelin, pegylated interferon alpha-2a , interferon alpha-2a, pegylated interferon alpha-2b, interferon alpha-2b, azacitidine, PEG-L-asparaginase, lenalidomide, gemtuzumab, hydrocortisone, interleukin-11, dexrazoxane, alemtuzumab, all-trans retinoic acid, ketoconazole, interleukin-2 , megestrol, immunoglobulin, nitrogen mustard, methylprednisolone, ibritgumomab tiuxetan, androgens, decitabine, hexamethylmelamine, bexarotene, tositumomab, arsenic trioxide, cortisone, editronate, mitotane, cyclosporine, liposomal daunorubicin, edwina-asparaginase, strontium 8 9, casopitant, netupitant, receptor antagonist NK-1, palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, alprazolam, haloperidol, droperidol, dronabinol, dexamethasone, methylprednisolone, prochlorperazine, granisetron, ondansetron, dolasetron, tropisetron, pegfilgrastim, erythropoietin, epoetin alfa, darb Epoetin alfa and mixtures thereof .

[00337] In any aspect or embodiment described herein, the bioactive agent or additional anticancer agent is a chemotherapy drug or biologic that targets epidermal growth factor receptors (e.g., an epidermal growth factor receptor inhibitor, such as at least one of gefitinib, erlotinib, neratinib, lapatinib, cetuximab, vandetanib, necitumab, osimertinib, or a combination thereof).

[00338] The term "anti-HIV antibody" or "additional anti-HIV antibody" includes, for example, nucleoside reverse transcriptase inhibitors (NRTIs), other non-nucleoside reverse transcriptase inhibitors (i.e., those not representative of the present invention), inhibitors proteases, fusion inhibitors, among others, typical compounds of which may include, for example, 3TC (lamivudine), AZT (zidovudine), (-)-FTC, ddI (didanosine), ddC (zalcitabine), abacavir (ABC), tenofovir (PMPA) ), D-D4FC (reverset), D4T (stavudine), rativir, L-FddC, L-FD4C, NVP (nevirapine), DLV (delavirdine), EFV (efavirenz), SQVM (saquinavir mesylate), RTV (ritonavir), IDV (indinavir), SQV (saquinavir), NFV (nelfinavir), APV (amprenavir), LPV (lopinavir), fusion inhibitors such as T20, among others, fuseon and mixtures thereof, including anti-HIV compounds currently in clinical trials testing or under development.

[00339] Other anti-HIV agents that may be used in conjunction with the compounds of the invention include, for example, other NNRTIs (i.e., other than the NNRTIs of the invention) that may be selected from the group consisting of nevirapine (BI-R6-587), delavirdine (U-90152S/T), efavirenz (DMP-266), UC-781 (N-[4-chloro-3-(3-methyl-2- butenyloxy)phenyl]-2-methyl3-furancarbothiamide), etravirine (TMC125), trovirdine (Ly300046.HCl), MKS-442 (emivirine, coactinone), HI-236, HI-240, HI-280, HI-281, rilpivirine (TMC-278), MSC-127, HBY 097, DMP266, baicalin (TJN-151) ADAM-II (methyl 3',3'-dichloro-4',4''-dimethoxy-5',5''- bis(methoxycarbonyl)-6,6-diphenylhexenoate), methyl 3-bromo-5-(1-5-bromo-4-methoxy-3-(methoxycarbonyl)phenyl)hept-1-enyl)-2-methoxybenzoate (analogue alkenyldiarylmethane, Adam's analogue), (5-chloro-3-(phenylsulfinyl)-2'-indolecarboxamide), AAP-VHAR (U-104489 or PNU-104489), capravirine (AG-1549, S-1153), atevirdine (U -87201E), aurine tricarboxylic acid (SD-095345), 1-[(6-cyano-2-indolyl)carbonyl]-4-[3-(isopropylamino)-2-pyridinyl]piperazine, 1-[5-[[ N-(methyl)methylsulfonylamino]-2-indolylcarbonyl-4-[3-(isopropylamino)-2-pyridinyl]piperazine, 1-[3-(ethylamino)-2-[pyridinyl]-4-[(5-hydroxy- 2-indolyl)carbonyl]piperazine, 1-[(6-formyl-2-indolyl)carbonyl]-4-[3-(isopropylamino)-2-pyridinyl]piperazine, 1-[[5-(methylsulfonyloxy)-2- indolyl)carbonyl]-4-[3-(isopropylamino)-2-pyridinyl]piperazine, U88204E, bis(2-nitrophenyl)sulfone (NSC 633001), calanolide A (NSC675451), calanolide B, 6-benzyl-5-methyl -2-(cyclohexyloxy)pyrimidin-4-one (DABO-546), DPC 961, E-EBU, E-EBU-dm, E-EPSeU, E-EPU, foscarnet (foscavir), HEPT (1-[(2 -hydroxyethoxy) methyl]-6-(phenylthio)thymine), HEPT-M (1-[(2-hydroxyethoxy)methyl]-6-(3-methylphenyl)thio)thymine), HEPT-S (1-[(2 -hydroxyethoxy)methyl]-6-(phenylthio)-2-thiothimine), inophyllum P, L-737,126, mishelamine A (NSC650898), mishelamine B (NSC649324), mishelamine F, 6-(3,5-dimethylbenzyl)-1 -[(2-hydroxyethoxy)methyl]-5-isopropyluracil, 6-(3,5-dimethylbenzyl)-1-(ethoxymethyl)-5-isopropyluracil, NPPS, E-BPTU (NSC 648400), oltipraz (4-methyl- 5-(pyrazinyl)-3H-1,2-dithiol-3-thione), N-{2-(2-chloro-6-fluorophenethyl]-N'-(2-thiazolyl)thiourea (PETT C1, derivative F) , N-{2-(2,6-difluorophenethyl]-N'-[2-(5-bromopyridyl)]thiourea {PETT derivative), N-{2-(2,6-difluorophenethyl]-N'-[2 -(5-methylpyridyl)]thiourea {pyridyl derivative of PETT), N-[2-(3-fluorofuranyl)ethyl]-N'-[2-(5-chloropyridyl)]thiourea, N-[2-(2-fluoro) -6-ethoxyphenethyl)]-N'-[2-(5-bromopyridyl)]thiourea, N-(2-phenethyl)-N'-(2-thiazolyl)thiourea(LY-73497), L-697,639, L- 697.593, L-697.661, 3-[2-(4,7-difluorobenzoxazol-2-yl)ethyl}-5-ethyl-6-methyl(pipridin-2(1H)-thione (2-pyridinone derivative)), 3 -[[(2-methoxy-5,6-dimethyl-3-pyridyl)methyl]amine-5-ethyl-6-methyl(pipridine-2(1H)-thione, R82150, R82913, R87232, R88703, R89439 (loviride ), R90385, S-2720, sodium suramin, TBZ (thiazolobenzimidazole, NSC 625487), thiazoloisoindol-5-one, (+)(R)-9b-(3,5-dimethylphenyl-2,3-dihydrothiazolo[2,3 -a]isoindol-5(9bH)-one, tivirapine (R86183), UC-38 hUC-84, among others.

[00340] The term "pharmaceutically acceptable salt" is used throughout the specification to refer, where applicable, to a salt form of one or more compounds described herein that are presented to increase the solubility of the compound in the gastric juices of the patient, the gastrointestinal tract, to promote dissolution and bioavailability of compounds. Pharmaceutically acceptable salts include salts derived from pharmaceutically acceptable inorganic or organic bases and acids, where applicable. Suitable salts include those derived from alkali metals such as potassium and sodium, alkaline earth metals such as calcium, magnesium and ammonium salts, among a variety of other acids and bases well known in the pharmaceutical art. Sodium and potassium salts are particularly preferred as phosphate neutralization salts according to the present invention.

[00341] The term "pharmaceutically acceptable derivative" is used throughout this specification to refer to any pharmaceutically acceptable form of a prodrug (such as an ester, an amide of another prodrug group) that, when administered to a patient, provides, directly or indirectly, the compound of the invention or an active metabolite of the compound of the invention.

General approach to synthesis

[00342] The implementation and optimization of the synthesis of bifunctional molecules, as described herein, can be carried out in a stepwise or modular manner. For example, identifying compounds that bind to target molecules may involve high- or medium-throughput screening campaigns if suitable ligands are not immediately available. Initial ligands often require iterative development and optimization cycles to improve suboptimal aspects, as determined by data from appropriate in vitro and pharmacological and/or ADMET, Absorption, Distribution, Metabolism, Excretion, and Toxicity testing (ADMET). and Toxicity). Part of the optimization/SAR campaign could be to investigate ligand positions that are resistant to substitution and that may be suitable sites for attachment of the linker chemistry previously mentioned herein. Where crystallographic or NMR (nuclear magnetic resonance) structural data are available, they can be used to guide such synthesis.

[00343] In a very similar way, ligands for E3 ligase, that is, ULM/ILM/VLM/CLM/ILM, can be identified and optimized.

[00344] With PTM and ULM (eg, ILM, VLM, CLM and/or ILM) available, one skilled in the art can use known synthetic methods to combine them with or without a linker moiety. Linker moieties can be synthesized with varying composition, length and flexibility and functionalized such that PTM and ULM groups can be sequentially attached to the distal ends of the linker. Thus, a library of bifunctional molecules can be implemented and profiled in in vitro and in vivo pharmacological studies, as well as in ADMET/PK studies. As with the PTM and ULM groups, the final bifunctional molecules can be subjected to iterative development and optimization cycles to identify molecules with desirable properties.

[00345] In some cases, protecting group and/or functional group interconversion (FGI) strategies may be required to facilitate the preparation of the desired materials. Such chemical processes are well known to those skilled in the field of synthetic organic chemistry, and many of them can be found in texts such as "Greene's Protective Groups in Organic Synthesis" by Peter G.M. Wuts and Theodora W. Greene (Wiley), and “Organic Synthesis: The Disconnection Approach” by Stuart Warren and Paul Wyatt (Wiley).

[00346] Abbreviations:

[00347] DIEA or DIPEA: diisopropylethylamine

[00348] DMF: N,N-dimethylformamide

[00349] DMSO: dimethyl sulfoxide

[00350] EDCI: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide

[00351] HOBt: hydroxybenzotriazole

[00352] Exemplary synthesis of intermediates.

[00353] Examples of the synthesis of intermediates that can be used for the synthesis of compounds according to the invention are described in Schemes A to I.

[00354] Diagram A.

[00355] A compound of formula I, which is commercially available or readily prepared using standard reaction methods known to those skilled in the art, can be reacted with a compound of formula II under conditions suitable for a nucleophilic aromatic substitution reaction, for example using a suitable base, such as diisopropylethylamine, in a suitable solvent such as N,N-dimethylpyrrolidone. In this case, Y represents a primary or secondary amine, which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring, or carboxylate; PG is H or a suitable optional protecting group including, but not limited to, tert-butoxycarbonyl when Y is a primary or secondary amine, or tert-butyl when Y is a carboxylate; L is an optional linker; Nu represents a suitable nucleophilic group such as O, or a primary or secondary amine, which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; X represents a suitable substituted group such as fluoride or chloride; and Z represents CH ₂ or C=O. In cases where III contains the optional protecting group PG, the PG can be removed under suitable conditions, for example hydrochloric acid in 1,4-dioxane if PG-Y is tert-butoxycarbonylamino, or trifluoroacetic acid in dichloromethane if PG-Y is tert-butyl carboxylate to obtain the compound of formula IV. In cases where PG is H, it is clear that III and IV are identical structures.

[00356] Circuit B.

[00357] Alternatively, a compound of formula V that is commercially available or readily prepared using standard reaction methods known to one skilled in the art can be reacted with a compound of formula VI to produce a compound of formula VII. In this case, PG, Y and L have the meanings shown in diagram A; Z is CH ₂ and Nu is O. LG may be a suitable substituted group such as tosylate, bromide or iodide, in which case the reaction conditions are those of nucleophilic substitution, for example using a suitable base such as potassium carbonate, and a suitable solvent such as N,N-dimethylformamide. Alternatively, LG may be H, in which case the reaction conditions correspond to the Mitsunobu reaction conditions, for example triphenylphosphine and diisopropyl azodicarboxylate. The compound of formula VII can then be converted to the compound of formula IV using conditions suitable for closing the imide ring and concomitant removal of PG if necessary, for example p-toluenesulfonic acid or benzenesulfonic acid in acetonitrile at 80°C.

[00358] Diagram C.

[00359] Alternatively, a compound of formula V may be reacted with a compound of formula VIII to produce a compound of formula III. In this case, Z is C=O and all other groups are as indicated in Scheme B. LG may be a suitable substitutable group such as a tosylate, in which case the reaction conditions are those of nucleophilic substitution, for example using a suitable base, such as potassium carbonate, and a suitable solvent such as N,N-dimethylformamide. The compound of formula III can then be converted to a compound of formula IV using the conditions described in Scheme A. Compounds of formula IV can be further converted into additional compounds of formula IV, for example, by manipulating the functional group on linker L using methods known to those skilled in the art field of technology. For example, if L contains an alkene, the alkene can be reduced under hydrogenation conditions, eg H ₂ , palladium on carbon in methanol, to give the corresponding alkane.

[00360] Diagram D.

[00361] A compound of formula VIII can react with a compound of formula IX to form compounds of formula X under amide-forming conditions, for example hydroxybenzotriazole, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and diisopropylethylamine in N,N-dimethylformamide or 1- [bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate and diisopropylethylamine or triethylamine in dichloromethane. In this case, Z represents an optional substituent, for example H, methyl or hydroxymethyl, Y represents a primary or secondary amine, which may be optionally substituted, or cyclized into a 4-8 membered heterocyclic ring, or is a carboxylate; PG represents a suitable optional protecting group including, but not limited to, tert-butoxycarbonyl if Y is a primary or secondary amine, or methyl if Y is a carboxylate; and L is an optional linker. The compound of formula X can then be converted to a compound of formula XI by removing the PG under suitable conditions, for example hydrochloric acid in 1,4-dioxane or dichloromethane if PG-Y is t-butoxycarbonylamino, or sodium hydroxide or lithium hydroxide in water, mixed with tetrahydrofuran and/or methanol, or hydrochloric acid in 1,4-dioxane if PG-Y is methyl carboxylate.

[00362] Diagram E.

[00363] A compound of formula XII can react with a compound of formula V to produce a compound of formula XIII. In this case, LG may be a suitable substituted group such as a tosylate, bromide or iodide; L is an optional linker; Y is a primary or secondary amine which may be optionally substituted or cyclized to form a 4-8 membered heterocyclic ring, or carboxylate; PG is H or a suitable optional protecting group including, but not limited to, tert-butoxycarbonyl when Y is a primary or secondary amine, or tert-butyl when Y is a carboxylate; or, alternatively, PG-Y together represents LG', a suitable substitutable group which may be the same or different from LG. The reaction conditions for preparing the compound of formula XIII correspond to the conditions of nucleophilic substitution, for example using a suitable base such as potassium carbonate and a suitable solvent such as N,N-dimethylformamide, at a temperature such as 80°C. PG can be removed under suitable conditions, for example hydrochloric acid in 1,4-dioxane if PG-Y is t-butoxycarbonylamino, or trifluoroacetic acid in dichloromethane if PG-Y is t-butyl carboxylate; or, alternatively, if PG-Y together are LG', they can be treated with a nucleophile, for example a primary amine in ethanol, in which case the Y in XIV becomes a secondary amine to give the compound of formula XIV. In cases where PG is H, it is clear that XIII and XIV are identical structures. If necessary, mixtures of enantiomers or diastereomers of any compounds XIII or XIV can be separated into their constituent enantiomers or diastereomers using methods known to one skilled in the art, including, but not limited to, preparative high performance liquid chromatography or preparative supercritical liquid chromatography.

[00364] Diagram F.

[00365] A compound of formula XV can react with compound V (readily prepared using standard reaction methods known to those skilled in the art) to produce a compound of formula XVI under nucleophilic substitution conditions, for example using a suitable base such as potassium carbonate, in a suitable solvent such as N,N-dimethylformamide. In this case, PG, Y, L and LG in compound V have the meanings indicated in diagram B; a PG' represents a suitable ester protecting group, for example methyl, ethyl or tert-butyl. Compounds of formula XVI can be converted to a compound of formula XVIII by treatment with a reagent suitable for removing PG', for example sodium hydroxide or lithium hydroxide in methanol and water if PG' is methyl or ethyl, or trifluoroacetic acid if PG' is tert -butyl. Compound XVII can react with a compound of formula XVIII wherein Z is an optional substituent, for example H, methyl or hydroxymethyl, and R is an optional substituent to produce compounds of formula XIX under amide formation conditions, for example 1-hydroxybenzotriazole, N-(3 -dimethylaminopropyl)-ethylcarbodiimide, diisopropylethylamine, N,N-dimethylformamide. The compound of formula XIX can be converted to the compound of formula XX by removing the PG under suitable conditions, for example hydrochloric acid in 1,4-dioxane if PG-Y is tert-butoxycarbonylamino, or trifluoroacetic acid in dichloromethane if PG-Y is tert -butyl carboxylate; or alternatively, if PG-Y together constitute a suitable substituent group, which may be the same or different from LG, they can be treated with a nucleophile, such as a primary amine in ethanol, in which case the Y in XX becomes a secondary amine. In cases where PG is H, it is clear that XIX and XX are identical structures. If necessary, mixtures of enantiomers or diastereomers of any compounds XVI, XVII, XIX or XX can be separated into their constituent enantiomers or diastereomers using methods known to one skilled in the art, including, but not limited to, preparative high performance liquid chromatography or preparative supercritical liquid chromatography.

[00366] Circuit G.

[00367] A compound of formula XXI can react with a compound of formula I under conditions suitable for coupling amides, for example 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine -3-oxide hexafluorophosphate and diisopropylethylamine in N,N-dimethylformamide to give the compound of formula XXII. In this case, Y represents a primary or secondary amine, which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; PG represents H or a suitable optional protecting group including, but not limited to, tert-butoxycarbonyl; L is an optional linker; and Nu is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring. In cases where XXII contains the optional protecting group PG, the PG can be removed under suitable conditions, for example hydrochloric acid in 1,4-dioxane if PG-Y is t-butoxycarbonylamino, to give the compound of formula XXIII. In cases where PG is H, it is clear that XXII and XXIII are identical structures.

[00368] Scheme N.

[00369] A compound of formula XXIV can react with a compound of formula V to produce a compound of formula XXV. In this case, LG may be a suitable substituted group such as a tosylate, bromide or iodide; L is an optional linker; Y is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring, and PG is H or a suitable optional protecting group including, but not limited to, benzyloxycarbonyl or 2-(trimethylsilyl)ethoxycarbonyl; or, alternatively, PG-Y together represents LG', a suitable substituent group which may be the same or different from LG; a IAP is either:

or

[00370] Reaction conditions correspond to those of nucleophilic substitution, for example, using a suitable base such as potassium carbonate and a suitable solvent such as N,N-dimethylformamide or acetonitrile, at a temperature such as 70-80°C. PG can be removed under suitable conditions to give a compound of formula XXVI, for example hydrogen, palladium on carbon if PG-Y is benzyloxycarbonylamino, or tetra-n-butylammonium fluoride in tetrahydrofuran if PG-Y is 2-(trimethylsilyl)ethoxycarbonylamino; or, alternatively, if PG-Y together are LG', they can be treated with a nucleophile, for example a primary amine in ethanol at 60°C, in which case the Y in XXVI becomes a secondary amine. In cases where PG is H, it is clear that XXV and XXVI are identical structures.

[00371] Scheme I.

[00372] Alternatively, a compound of formula XXIV, as indicated in scheme H, can react with another compound of formula V, where PG is a suitable protecting group such as tetrahydropyranyl, Y is O, L is an optional linker, and LG is a suitable substituted group such as tosylate, bromide or iodide, under conditions of nucleophilic substitution, for example potassium carbonate and potassium iodide in N,N-dimethylformamide at 70° C. to give the compound of formula XXVII. The compound of formula XXVII can be deprotected, for example, using pyridinium p-toluenesulfonate in methanol to give compound XXVIII. The compound of formula XXVIII can be converted to the compound of formula XXIX by treatment with, for example, p-toluenesulfonyl chloride, triethylamine, N,N-dimethylaminopyridine in dichloromethane. Finally, the compound of formula XXIX can be treated with a nucleophile, such as a primary amine in ethanol at 60°C, to convert it to the compound of formula XXVI, as indicated in scheme H.

[00373] General synthesis schemes:

[00374] The compounds of the present invention can be synthesized as shown in Schemes 1 to 19 using the intermediate structures described in Schemes A to I.

[00375] Scheme 1.

[00376] A compound of formula XXX can react with a compound of formula IV under amide coupling conditions, for example 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine 3-oxidehexafluorophosphate and diisopropylethylamine in N,N-dimethylformamide or 1-hydroxybenzotriazole and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide, diisopropylethylamine in N,N-dimethylformamide to give a compound of formula XXXI.

[00377] In this case is aryl, heteroaryl, cycloalkyl or heterocycloalkyl; each X is independently CH or N; R ₁ represents one or more independent alkyls, alkoxys, phenyls or naphthalenes, each of which may be independently substituted with OH, H and/or halogen; R ₂ represents an optionally substituted alkylamide, alkenylamide, urea or a suitable protecting group such as t-butoxycarbonyl; R ₃ represents an optional substituent; L' is an optional linker; W represents a carboxylic acid; Y is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; and L, Nu and Z have the values indicated in one of the diagrams A, B or C.

[00378] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula XXXI under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine.

[00379] Scheme 2.

[00380] A compound of formula XXX can also react with a compound of formula IV under amide binding conditions, for example, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridin- Hexafluorophosphate 3-oxide and triethylamine in N,N-dimethylformamide to give the compound of formula XXXII.

[00381] In this case, R ₂ represents H; L'-W together represent H or form an optional surrogate; , X, R ₁ , R ₃ have the values indicated in Scheme 1; Y represents a carboxylic acid; a L, Nu and Z have the values indicated in one of the diagrams A, B or C.

[00382] Scheme 3.

[00383] A compound of formula XXX can also react with a compound of formula XI under amide coupling conditions, for example, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridin- Hexafluorophosphate 3-oxide and diisopropylethylamine in N,N-dimethylformamide to give the compound of formula XXXIII.

[00384] In this case , X, R ₁ , R ₂ , R ₃ , L', and W have the meanings indicated in Scheme 1; Y is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; a L and Z have the meanings shown in diagram D.

[00385] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula XXXIII under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature.

[00386] Scheme 4.

[00387] A compound of formula XXX can also react with a compound of formula XI under amide coupling conditions, for example, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridin- Hexafluorophosphate 3-oxide and triethylamine in N,N-dimethylformamide to give the compound of formula XXXIV.

[00388] In this case, R ₂ represents H; L'-W together represent H or form an optional surrogate; , X, R ₁ , R ₃ have the values indicated in Scheme 1; Y represents a carboxylic acid; a L and Z have the meanings shown in diagram D.

[00389] Diagram 5.

[00390] A compound of formula XXX can also react with a compound of formula XXVI under amide coupling conditions, for example, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridin- Hexafluorophosphate 3-oxide and diisopropylethylamine in N,N-dimethylformamide to give the compound of formula XXXV.

[00391] In this case , X, R ₁ , R ₂ , R ₃ , L' and W have the meanings indicated in Scheme 1; Y is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; a L and IAP have the meanings indicated in one of the diagrams H or I.

[00392] The tert-butoxycarbonyl group contained in the IAP structure can then be removed under suitable conditions, for example, hydrochloric acid in 1,4-dioxane or difluoroacetic acid in dichloromethane, to give various compounds of formula XXXV, where the IAP structure is either :

or

[00393] Diagram 6.

[00394] A compound of formula XXX can also react with a compound of formula XXIII under amide coupling conditions, for example, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridin- Hexafluorophosphate 3-oxide and diisopropylethylamine in N,N-dimethylformamide to give the compound of formula XXXVI.

[00395] In this case , X, R ₁ , R ₂ , R ₃ , L' and W have the meanings indicated in Scheme 1; Y is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; a L and Nu have the meanings shown in diagram G.

[00396] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula XXXVI under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature.

[00397] Diagram 7.

[00398] A compound of formula XXX can also react with a compound of formula XIV under amide coupling conditions, for example, hydroxybenzene triazole, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and diisopropylethylamine in N,N-dimethylformamide to produce a compound of formula XXXVII.

[00399] In this case , X, R ₁ , R ₂ , R ₃ , L' and W have the meanings indicated in Scheme 1; Y is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; a L has the values indicated in diagram E.

[00400] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula XXXVII under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature. If necessary, mixtures of enantiomers or diastereomers of any compounds XXXVII can be separated into their constituent enantiomers or diastereomers using methods known to one skilled in the art, including, but not limited to, preparative high performance liquid chromatography or preparative supercritical liquid chromatography.

[00401] Diagram 8.

[00402] A compound of formula XXX can also react with a compound of formula I under amide coupling conditions, for example, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridin- Hexafluorophosphate 3-oxide and triethylamine in N,N-dimethylformamide to give the compound of formula XXXIV.

[00403] In this case, R ₂ represents H; L'-W together represent H or form an optional surrogate; , X, R ₁ , R ₃ have the values indicated in Scheme 1; Nu-H is a primary or secondary amine; L is an optional linker; Y is a carboxylate and PG is a suitable carboxylic acid protecting group including, but not limited to, methyl or ethyl. The compound of formula XXXVIII can then be converted to the compound of formula XXXIX under conditions suitable for removing PG, for example lithium hydroxide in water and a mixture of methanol and/or tetrahydrofuran if the PG is methyl or ethyl. Finally, a compound of formula XXXIX can react with a compound of formula VIII under conditions suitable for coupling amides, for example, hydroxybenzene triazole, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and diisopropylethylamine in 1 N,N-dimethylformamide to give a compound of formula XXXIV. In this case, Z has the value indicated in diagram D.

[00404] Diagram 9.

[00405] A compound of formula XXX may also react with a compound of formula XX under amide coupling conditions, for example, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridin- Hexafluorophosphate 3-oxide and diisopropylethylamine in N,N-dimethylformamide to give the compound of formula XXXVIII.

[00406] In this case , X, R ₁ , R ₂ , R ₃ , L' and W have the meanings indicated in Scheme 1; Y is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; a L, Z and R have the meanings shown in diagram F.

[00407] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula XXXVIII under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature. If necessary, mixtures of enantiomers or diastereomers of any compounds XXXVIII can be separated into their constituent enantiomers or diastereomers using methods known to one of ordinary skill in the art, including, but not limited to, preparative high performance liquid chromatography or preparative supercritical liquid chromatography.

[00408] Diagram 10.

[00409] A compound of formula XXX can also react with a compound of formula XI under nucleophilic aromatic substitution reaction conditions, for example diisopropylethylamine in isopropanol at 115°C with microwave heating to produce a compound of formula XXXIX.

[00410] In this case , X, R ₁ , R ₂ , and R ₃ are as defined in Scheme 1, L' is absent, W represents a suitable substituted group such as fluoride or chloride; Y is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; a L and Z have the meanings shown in Scheme D. In cases where R ₁ is an aryl or heteroaryl chloride, bromide or iodide, the compound of formula XXXIX can be further converted to another compound of formula XXXIX, for example, under the conditions of the Suzuki coupling reaction with arylboronic acid or ester with catalytic (2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II ) methanesulfonate, potassium phosphate in water and tetrahydrofuran at 60°C. In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula XXXIX under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature.

[00411] Diagram 11.

[00412] A compound of formula XXX can also react with a compound of formula XL under nucleophilic aromatic substitution reaction conditions, for example, sodium hydride in N,N-dimethylformamide at room temperature to produce a compound of formula XLI.

[00413] In this case , X, R ₁ , R ₂ , and R ₃ are as defined in Scheme 1, L' is absent, W represents a suitable substituted group such as fluoride or chloride; L'' represents a linker; and Y' is either CH ₂ OH or a terminal, suitably protected (eg tert-butoxycarbonyl) primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring.

[00414] In cases of XLI, where Y' is CH ₂ OH, it can be converted to its corresponding aldehyde under oxidizing conditions, such as Swern or Dess-Martin oxidation, to give compound XLII, where Y'' is CHO. In cases of XLI where Y' is a terminal, suitably protected primary or secondary amine, it can be converted to the corresponding unprotected amine using suitable conditions, for example trifluoroacetic acid in dichloromethane at room temperature when the protecting group is tert-butoxycarbonyl , to obtain compound XLII, where Y'' represents the amine of Y' without a protecting group. The compound of formula XLII can then be treated with a compound of formula XI under reductive amination conditions, for example sodium triacetoxyborohydride and triethylamine in dichloromethane or sodium cyanoborohydride, sodium acetate and acetic acid in dichloromethane and methanol, to give the compound of formula XLIII. In this case, L and Z have the meanings indicated in diagram D. In cases where Y'' in XLII represents CHO, Y in XI represents a primary or secondary amine, which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; a Y'' in XLIII becomes CH ₂ . In cases where Y'' in XLII represents a primary or secondary amine, which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring, Y in XI is CHO; a Y in XLIII becomes CH ₂ .

[00415] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula XLIII under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature.

[00416] Diagram 12.

[00417] A compound of formula XXX can react with a compound of formula IV under reductive amination conditions, such as sodium triacetoxyborohydride and triethylamine in dichloromethane or sodium cyanoborohydride, sodium acetate and acetic acid in dichloromethane and methanol, to produce a compound of formula XXXI.

[00418] In this case , R ₁ , R ₂ , R ₃ and L' have the meanings indicated in Scheme 1; W is CHO; Y is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; a Nu, L and Z have the values indicated in one of the diagrams A, B or C.

[00419] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula XLIV under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine.

[00420] Diagram 13.

[00421] A compound of formula XXX can react with a compound of formula XIV under reductive amination conditions, such as sodium triacetoxyborohydride and triethylamine in dichloromethane or sodium cyanoborohydride, sodium acetate and acetic acid in dichloromethane and methanol, to produce a compound of formula XLV.

[00422] In this case , X, R ₁ , R ₂ , R ₃ , and L' have the meanings indicated in Scheme 1; W is CHO; Y is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; a L has the values indicated in diagram E.

[00423] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula XLV under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature. If necessary, mixtures of enantiomers or diastereomers of any XLV compounds can be separated into their constituent enantiomers or diastereomers using methods known to one of ordinary skill in the art, including, but not limited to, preparative high performance liquid chromatography or preparative supercritical liquid chromatography.

[00424] Diagram 14.

[00425] A compound of formula XXX can also react with a compound of formula XXVI under reductive amination conditions, for example, sodium triacetoxyborohydride and triethylamine in dichloromethane or sodium cyanoborohydride, sodium acetate and acetic acid in dichloromethane and methanol, to give a compound of formula XLVI.

[00426] In this case , X, R ₁ , R ₂ , R ₃ and L' have the meanings indicated in Scheme 1; W is CHO; Y is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; a L and IAP have the meanings indicated in one of the diagrams H or I.

[00427] The tert-butoxycarbonyl group contained in the IAP structure can then be removed under suitable conditions, for example, hydrochloric acid in 1,4-dioxane or difluoroacetic acid in dichloromethane, to give various compounds of formula XLVI, where the IAP structure is either :

or

[00428] Diagram 15.

[00429] A compound of formula XXX can also react with a compound of formula XXIII under reductive amination conditions, such as sodium triacetoxyborohydride and triethylamine in dichloromethane or sodium cyanoborohydride, sodium acetate and acetic acid in dichloromethane and methanol, to produce a compound of formula XLVII.

[00430] In this case , X, R ₁ , R ₂ , R ₃ and L', have the meanings indicated in Scheme 1; W is CHO; Y is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; a L and Nu have the meanings shown in diagram G.

[00431] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula XXXVII under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature.

[00432] Diagram 16.

[00433] A compound of formula XLVIII can react with a compound of formula V to produce a compound of formula XLIX. In this case, LG may be a suitable substituent group such as a tosylate, bromide or iodide; L is an optional linker; Y is a primary or secondary amine which may be optionally substituted or cyclized into a 4-8 membered heterocyclic ring; PG is a suitable protecting group including, but not limited to, tert-butoxycarbonyl; and PG' represents a suitable protecting group including, but not limited to, 2-(trimethylsilyl)ethoxy]methyl. PG and PG' can then be removed simultaneously using suitable conditions such as hydrochloric acid in 1,4-dioxane to give a compound of formula L. The compound of formula L can then be treated with a compound of formula XXX under reductive amination conditions such as sodium triacetoxyborohydride and triethylamine in dichloromethane or sodium cyanoborohydride, sodium acetate and acetic acid in dichloromethane and methanol to give the compound of formula LI.

[00434] In this case , X, R ₁ , R ₂ , R ₃ and L' have the meanings indicated in Scheme 1; a W represents CHO. In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula LI under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature.

[00435] Diagram 17.

[00436] A compound of formula XXX can also react with a compound of formula LII under nucleophilic aromatic substitution reaction conditions, for example, sodium hydride in tetrahydrofuran at a temperature of 20 to 50°C to produce a compound of formula LIII. In this case, X and R ₃ have the values indicated in diagram 1; is an optionally substituted aliphatic cyclic amine; R1 represents a suitable amino protecting group for example tert-butoxycarbonyl; R ₂ represents a suitable protecting group, for example benzyloxycarbonyl; L' is missing; W represents a suitable substituted group such as fluoride or chloride; L'' represents a linker; and PG is a suitable alcohol protecting group, for example tetrahydropyranyl. The compound of formula LIII can be converted to the compound of formula LIV under conditions suitable for removing certain protecting groups, for example trifluoroacetic acid in dichloromethane when R ₁ is t-butoxycarbonyl and PG is tetrahydropyranyl; in this case, R ₁ in LIV becomes H. Such a compound of formula LIV can be converted into another compound of the general formula LIV using conditions known to one skilled in the art. For example, taking compound LIV, where R ₁ is H, and treating it with an aryl halide under conditions suitable for Hartwig-Buchwald amination, for example (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)[2 -(2'-amino-1,1'-biphenyl)]palladium(II) methanesulfonate and cesium carbonate in 1,4-dioxane at 90°C, give a compound of formula LIV, where R ₁ represents, for example, phenyl, naphthalene or heteroaryl, which may be independently substituted multiple times with OH, CN, alkyl and/or halogen. Further conversions of a LIV compound to another LIV compound can be achieved, for example, by changing the identity of the R ₂ group. For example, in the case where R ₂ represents benzyloxycarbonyl, it can be treated, for example, with H ₂ , palladium on carbon in methanol to obtain the compound LIV, where R ₂ represents H. Said compound of formula LIV, where R ₂ represents H, can be converted, for example, by treatment with di-tert-butyl dicarbonate and triethylamine in dichloromethane to another compound of formula LIV, where R ₂ represents tert-butoxycarbonyl; or for example by treatment with isocyanatotrimethylsilane and triethylamine in tetrahydrofuran to another compound of formula LIV wherein R ₂ is C(O)NH ₂ . A suitable substituted compound of formula LIV can be converted to a compound of formula LV wherein PG is a suitable protecting group such as ethyl by treatment with a reagent such as ethyl diazoacetate and a catalyst such as rhodium(II) acetate in dichloromethane. The compound of formula LV can be converted to the compound of formula LVI using conditions suitable for ester hydrolysis, such as lithium hydroxide in water and tetrahydrofuran. Finally, a compound of formula LVI can react with a compound of formula VIII under conditions suitable for coupling amines, for example, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, 1-hydroxybenzotriazole and diisopropylethylamine in N,N-dimethylformamide to give compounds of formula LVII. In this case, Z represents an optional substituent, for example H, methyl or hydroxymethyl.

[00437] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula LVII under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature. Alternatively, said compound wherein R ₂ is H can then be converted to another compound of formula LVII under urea formation conditions. Typical conditions for such amide formations include, but are not limited to, isocyanatotrimethylsilane and triethylamine in N,N-dimethylformamide at room temperature.

[00438] Diagram 18.

[00439] Alternatively, the compound of formula LIV can be converted to the compound of formula LVIII by treatment with a suitable reagent such as p-toluenesulfonyl chloride, triethylamine and N,N-dimethylaminopyridine in dichloromethane.

[00440] In this case , R ₁ , R ₂ , R ₃ and L'' have the values indicated for LIV in Scheme 17; a LG is, for example, p-toluenesulfonyl chloride. The compound of formula LVIII can then react with a compound of formula VIII under conditions suitable for nucleophilic substitution, for example potassium carbonate in N,N-dimethylformamide at 50° C. to give the compound of formula LIX. In this case, Z represents C=O.

[00441] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula LIX under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature. Alternatively, said compound wherein R ₂ is H can then be converted to another compound of formula LIX under urea formation conditions. Typical conditions for such urea formations include, but are not limited to, isocyanatotrimethylsilane and triethylamine in N,N-dimethylformamide at room temperature.

[00442] Diagram 19.

[00443] Alternatively, a compound of formula LVIII can be converted to a compound of formula LX by treatment with a compound of formula XII under conditions suitable for an alkylation reaction, for example potassium carbonate in N,N-dimethylformamide at 80°C.

[00444] In this case , R ₁ , R ₂ , R ₃ and L'' have the values indicated for LIV in diagram 17.

[00445] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula LX under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature. Alternatively, said compound wherein R ₂ is H can then be converted to another compound of formula LX under urea formation conditions. Typical conditions for such urea formations include, but are not limited to, isocyanatotrimethylsilane and triethylamine in N,N-dimethylformamide at room temperature.

[00446] Diagram 20.

[00447] Alternatively, a compound of formula LVIII can be converted to a compound of formula LXI by treatment with an ammonia equivalent such as potassium phthalimide in N,N-dimethylformamide at 80° C. followed by treatment with, for example, hydrazine hydrate in ethanol at 70 °C.

[00448] In this case , R ₁ , R ₂ , R ₃ and L'' have the values indicated for LIV in diagram 17.

[00449] The compound of formula LXI can then react with the compound of formula II, under conditions suitable for nucleophilic aromatic substitution, diisopropylethylamine in dimethyl sulfoxide at 90°C to obtain the compound of formula LXII. In this case, Z represents C=O.

[00450] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as trifluoroacetic acid in dichloromethane if R ₂ is tert-butoxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula LXII under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature. Alternatively, said compound wherein R ₂ is H can then be converted to another compound of formula LXII under urea formation conditions. Typical conditions for such urea formations include, but are not limited to, isocyanatotrimethylsilane and triethylamine in N,N-dimethylformamide at room temperature.

[00451] Diagram 21.

[00452] Alternatively, a compound of formula LVIII can be reacted with a compound of formula VI to produce a compound of formula LXIII under conditions suitable for nucleophilic substitution, for example potassium carbonate in N,N-dimethylformamide at 80°C.

[00453] In this case , R ₁ , R ₂ , R ₃ and L'' have the values given for LIV in Scheme 17, and Z is CH ₂ .

[00454] The compound of formula LXIII can then be treated with an imide ring-closing reagent and concomitant removal of R ₂ in cases where R ₂ is tert-butoxycarbonyl, for example benzenesulfonic acid in acetonitrile under reflux. In cases where R ₂ is tert-butoxycarbonyl in LVIII, R ₂ becomes H in LXIV. The specified compound LXIV, where R ₂ represents H, can then be converted into another compound of formula LXIV under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature. Alternatively, said compound wherein R ₂ is H can then be converted to another compound of formula LXIV under urea formation conditions. Typical conditions for such urea formations include, but are not limited to, isocyanatotrimethylsilane and triethylamine in N,N-dimethylformamide at room temperature.

[00455] Diagram 22.

[00456] Alternatively, a compound of formula LVIII can be converted to a compound of formula LXV by treatment with a compound of formula XXIV under conditions suitable for an alkylation reaction, for example potassium carbonate in acetonitrile at 80°C.

[00457] In this case , R ₁ , R ₂ , R ₃ and L'' have the values given for LIV in Figure 17, and IAP is either:

or

[00458] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as H ₂ , palladium on carbon in methanol when R ₂ is benzyloxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula LXVI under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature.

[00459] The compound of formula LXV can subsequently be converted to the compound of formula LXVI by treatment with reagents suitable for removing the tert-butoxycarbonyl protecting group, for example trifluoroacetic acid in dichloromethane. In this case, IAP' is either:

or

[00460] Diagram 23.

[00461] A compound of formula LXI may alternatively react with a compound of formula XXI under amide coupling conditions, for example, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, 1-hydroxybenzotriazole and diisopropylethylamine in N,N-dimethylformamide with obtaining the compound of formula LXVII.

[00462] In this case , R ₁ , R ₂ , R ₃ and L'' have the values indicated for LIV in diagram 17.

[00463] In cases where R ₂ is a protecting group, the protecting group can be removed under suitable conditions, such as H ₂ , palladium on carbon in methanol when R ₂ is benzyloxycarbonyl. The specified compound, where R ₂ represents H, can then be converted into another compound of formula LXVII under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature.

[00464] Diagram 24.

[00465] Alternatively, a compound of formula LVIII can be converted to a compound of formula LXVIII by treatment with a compound of formula XLVIII under conditions suitable for an alkylation reaction, for example potassium carbonate in acetonitrile at 80°C.

[00466] In this case , R ₁ , R ₂ , R ₃ and L'' have the meaning given for LIV in Scheme 17, and PG' has the meaning given in Scheme 16. In cases where R ₂ is a protecting group, the protecting group may be removed under suitable conditions such as hydrogen chloride in 1,4-dioxane; in cases where PG' is 2-(trimethylsilyl)ethoxymethyl, PG' becomes H under these conditions. The specified compound, where PG' and R ₂ represent H, can then be converted into another compound of formula LXVIII under amide coupling conditions. Typical conditions for such amide formation include, but are not limited to: acryloyl chloride, 2,6-lutidine, dichloromethane, minus 78°C; or a carboxylic acid such as 2,2-dihydroxyacetic acid, and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine hexafluorophosphate 3-oxide and diisopropylethylamine in N, N-dimethylformamide at room temperature.

[00467] General synthesis scheme 1.

[00468] The compound of formula I (commercially available or readily prepared using standard reaction methods known to those skilled in the art) can react with compound II under amide-forming conditions, for example HOBt, EDCI, with a suitable base such as DIEA , and a suitable solvent, such as DMF, to obtain the compound of formula III. PG is a suitable protecting group, for example tert-butoxycarbonyl, R is H or an optional substituent, for example methyl, and Z is an optional substituent, for example H, methyl or hydroxymethyl. The compound of formula III can be converted to the compound of formula IV using conditions suitable for deprotection, for example hydrogen chloride in 1,4-dioxane in dichloromethane when PG is t-butoxycarbonyl. The compound of formula IV can then react with a compound of formula V under amide coupling conditions, eg, similar to those used to convert I and II to III, to produce a compound of formula VI. Compounds VI can then be converted to compounds VII under suitable deprotecting conditions, for example trifluoroacetic acid in dichloromethane. Compound VII can be converted to a compound of formula VIII under amide formation conditions, eg acryloyl chloride, 2,6-lutidine, dichloromethane.

[00469] General synthesis scheme 2.

[00470] A compound of formula IX can react with a compound of formula X to produce compounds of formula XI, wherein X is a suitable substituent group such as fluorine or chlorine, PG is a suitable protecting group such as tert-butoxycarbonyl, R is H or an optional substituent, eg methyl, and the reaction conditions are the same as for nucleophilic aromatic substitution, eg DIEA, DMSO, 80°C. A compound of formula XI can be converted to a compound of formula XII using conditions suitable for deprotection, for example hydrogen chloride in 1,4-dioxane in dichloromethane when PG is t-butoxycarbonyl. The compound of formula XI can then be reacted with the compound of formula V under amide coupling conditions, eg HOBt, EDCI, with a suitable base such as DIEA and a suitable solvent such as DMF to produce the compound of formula XIII. Compound XIII can then be converted to Compound XIV under suitable deprotection conditions, for example trifluoroacetic acid in dichloromethane. Compound XIV can be converted to the compound of formula XV under amide formation conditions, eg acryloyl chloride, 2,6-lutidine, dichloromethane.

[00471] Example of synthesis of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione

[00472] Step 1: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione

[00473] Sodium acetate (4.1 g, 49.4 mmol, 3.00 eq.), then the mixture was stirred at 25°C for 1 hour. 4-Hydroxyphthalic acid (3.0 g, 16.5 mmol, 1.00 eq.) was then added to the mixture, heated to 120°C and stirred for another 11 hours. The mixture was concentrated and then poured into water (20 ml) and then filtered. The crude product was purified by column chromatography (dichloromethane:methanol = 50:1 to 10:1) to give 2-(2,6-dioxo-3-piperidyl)-5-hydroxyisoindoline-1,3-dione (3.9 g , 14.3 mmol, 86% yield) as a colorless solid. LC/MS (liquid chromatography with mass spectrometry) (ESI (electrospray ionization, electrospray ionization)) m/z (mass/charge): 275 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 11.19-10.94 (m, 2H), 7.75 (d, J=8.0 Hz, 1H), 7.20-7.08 (m, 2H), 5.08 (dd, J=5.2, 12.8 Hz, 1H), 3.34 (br s, 1H), 2.95-2.81 (m, 1H), 2.64- 2.55 (m, 1H), 2.08-1.98 (m, 1H).

[00474] Synthesis example of (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazole) -5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide hydrochloride

[00475] Step 1: Preparation of tert-butyl (S)-(1-(4-bromophenyl)ethyl) carbamate

[00476] Place (1S)-1-(4-bromophenyl)ethan-1-amine (10.0 g, 49.98 mmol, 1.00 eq) in dichloromethane (100 mL) into a 250 mL round bottom flask. , triethylamine (10.0 g, 99.01 mmol, 2.00 eq.), di-tert-butyl dicarbonate (13.0 g, 59.63 mmol, 1.20 eq.). The resulting solution was stirred for 2 hours at room temperature. The resulting mixture was concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:10). The result was 15.0 g of tert-butyl-N-[(1S)-1-(4-bromophenyl)ethyl]carbamate as a white solid.

[00477] Step 2: Preparation of tert-butyl (S)-(1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamate

[00478] A solution of tert-butyl-N-[(1S)-1-(4-bromophenyl)ethyl]carbamate (15.0 g, 49.97) was placed in a 250 mL round bottom flask, purged and maintained under an inert nitrogen atmosphere. mmol, 1.00 eq.) in N,N-dimethylacetamide (100 ml), 4-methyl-1,3-thiazole (9.9 g, 99.84 mmol, 2.00 eq.), potassium acetate (9 .8 g, 99.86 mmol, 2.00 eq.), palladium (II) acetate (112.5 mg, 0.50 mmol, 0.01 eq.). The resulting solution was stirred for 2 hours at a temperature of 120°C. The reaction mixture was quenched by adding water (500 ml). The resulting solution was extracted with ethyl acetate (200 ml x 3), the organic layers were combined and concentrated in vacuo. The residue was applied to a silica gel column with a mixture of ethyl acetate/petroleum ether (1:5). The result was 7.5 g (47%) of tert-butyl-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]carbamate as white solid matter. LC/MS (ESI) m/z: 319.13 [M+Na] ⁺ .

[00479] Step 3: Preparation of (S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethan-1-amine hydrochloride

[00480] A solution of tert-butyl-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]carbamate (7, 5 g, 23.55 mmol, 1.00 eq) in methanol (20 ml), bubbled with hydrogen chloride (gas) at room temperature. The resulting solution was stirred for 2 hours at room temperature. The resulting mixture was concentrated in vacuo. The result was 4.4 g (86%) of (1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethan-1-amine as a white solid.

[00481] Step 4: Preparation of tert-butyl (2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine- 1-carboxylate

[00482] Place (2S,4R)-1-[(tert-butoxy)carbonyl]-4-hydroxypyrrolidine-2-carboxylic acid (4.7 g, 20.32 mmol, 1.00 eq) into a 100 mL round bottom flask .) in N,N-dimethylformamide (20 ml), N-ethyl-N-isopropylpropan-2-amine (7.8 g, 60.35 mmol, 3.00 eq.), o-(7-azabenzotriazole-1 -yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.5 g, 30.26 mmol, 1.50 eq.), (1S)-1-[4-(4-methyl-1, 3-thiazol-5-yl)phenyl]ethan-1-amine (4.4 g, 20.15 mmol, 1.00 eq.). The resulting solution was stirred for 12 hours at room temperature. The reaction mixture was quenched by adding water (20 ml). The resulting solution was extracted with ethyl acetate (100 ml x 3), the organic layers were combined and dried in an oven under reduced pressure, and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:1). The result was 5.0 g (57%) tert-butyl (2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methyl-1,3-thiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carboxylate as a yellow solid. LC/MS (ESI) m/z: 432.15 [M+1] ⁺ .

[00483] Step 5: Preparation of (2S,4R)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide hydrochloride.

[00484] A solution of tert-butyl (2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methyl-1,3-thiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carboxylate (5.0 g, 11.59 mmol, 1.00 eq.) in methanol (200 ml), then the reaction mixture was bubbled with hydrogen chloride (gas) for 2 hours at room temperature. The resulting mixture was concentrated in vacuo. The result was 3.2 g (83%) of (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl] ethyl]pyrrolidine-2-carboxamide as a red solid.

[00485] Step 6: Preparation of tert-butyl ((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl) phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)carbamate

[00486] Place (2S)-2-[(tert-butoxy)carbonyl]amino-3,3-dimethylbutanoic acid (2.0 g, 8.65 mmol, 0.99 eq) in a 25 mL round bottom flask. N,N-dimethylformamide (30 ml). N-ethyl-N-isopropylpropan-2-amine (3.4 g, 3.00 eq.), o-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (5 .0 g, 1.50 eq), (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl ]pyrrolidine-2-carboxamide hydrochloride (3.2 g, 8.70 mmol, 1.00 eq.). The resulting solution was stirred for 12 hours at room temperature. The resulting solution was extracted with ethyl acetate (60 ml x 3) and washed with water (100 ml x 2). The organic layers were combined and dried, concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:3). The result was 4.0 g (84%) tert-butyl N-[(2S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methyl 1,3-thiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate as a yellow solid. LC/MS (ESI) m/z: 545.30 [M+1] ⁺ .

[00487] Step 7: Preparation of (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4- methylthiazol)-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide hydrochloride

[00488] A solution of tert-butyl-N-[(2S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4 -methyl-1,3-thiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate (4.0 g, 7.34 mmol, 1.00 eq.) in methanol (30 ml), then hydrogen chloride (gas) was bubbled through the reaction mixture for 2 hours at room temperature. The resulting mixture was concentrated in vacuo. The result was 3.5 g of (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4- methyl 1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide hydrochloride as a yellow solid. LC/MS (ESI) m/z: 445.05 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.99 (s, 1 H), 8.57-8.55 (d, J=7.8 Hz, 1 H), 8.01 (b , 3 N), 7.46-7.43 (d, J=8.4 Hz, 2 N), 7.39-7.37 (d, J=8.4 Hz, 2 N), 4.98 -4.90 (m, 1 N), 4.57-4.51 (m, 1 N), 4.34 (b, 1 N), 3.94-3.92 (m, 1 N), 3 .69-3.66 (m, 1 N), 3.53-3.49 (m, 1 N), 2.52 (s, 3 N), 2.10-2.07 (m, 1 N) , 1.83-1.81 (m, 1 N), 1.40-1.30 (m, 3 N), 1.03 (s, 9 N).

[00489] Example of the synthesis of 3-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)propanoic acid

[00490] Step 1: Preparation of 2-amino-4-bromo-5-chloro-3-fluorobenzoic acid

[00491] To a solution of 2-amino-4-bromo-3-fluorobenzoic acid (9.0 g, 38.5 mmol) in N,N-dimethylformamide (180 ml) was added N-chlorosuccinimide (6.2 g, 46. 2 mmol) and stirred at 70°C overnight. After cooling to room temperature, the reaction mixture was poured into water (100 ml). The precipitate was collected by filtration, and the filter cake was washed with water and dried in vacuo to give 2-amino-4-bromo-5-chloro-3-fluorobenzoic acid (9.0 g, 87%) as a yellow solid. LC/MS (ESI) m/z: 268.0 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 6.90 (br, 2H), 7.69 (d, J=2.0 Hz, 1H).

[00492] Step 2: Preparation of 7-bromo-6-chloro-8-fluoroquinazoline-2,4(1H,3H)-dione.

[00493] A mixture of 2-amino-4-bromo-5-chloro-3-fluorobenzoic acid (2.0 g, 7.5 mmol) and urea (2.2 g, 37.3 mmol) was mixed until smooth at room temperature temperature and stirred at 240°C for 2 hours. After cooling to 100°C, water (30 ml) was added and the resulting mixture was stirred at 100°C for 30 minutes. The precipitate was collected by filtration, the filter cake was washed with boiling water three times and dried in vacuum to obtain 7-bromo-6-chloro-8-fluoroquinazoline-2,4(1H,3H)-dione (2.1 g, 95%) as yellow solid. LC/MS (ESI) m/z: 293.6 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.82 (d, J=1.6 Hz, 1H), 11.65 (br, 2H).

[00494] Step 3: Preparation of 7-bromo-2,4,6-trichloro-8-fluoroquinazoline.

[00495] To a suspension of 7-bromo-6-chloro-8-fluoroquinazoline-2,4(1H,3H)-dione (500 mg, 1.7 mmol) in phosphorus oxychloride (7.5 ml) was added N-ethyl- N-isopropylpropan-2-amine (550 mg, 4.3 mmol) and reflux overnight. The volatile components were evaporated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (2% ethyl acetate in hexanes) to give 7-bromo-2,4,6-trichloro-8-fluoroquinazoline (400 mg, 71%) in as a yellow solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.21 (d, J=2.0 Hz, 1H).

[00496] Step 4: Preparation of tert-butyl 4-(7-bromo-2,6-dichloro-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate

[00497] To a solution of 7-bromo-2,4,6-trichloro-8-fluoroquinazoline (400 mg, 1.2 mmol) and triethylamine (364 mg, 3.6 mmol) in 1,4-dioxane (6 ml) tert-butylpiperazine-1-carboxylate (236 mg, 1.27 mmol) was added at room temperature and stirred at 50°C for 30 minutes. The reaction mixture was quenched with water (15 ml) and extracted with dichloromethane (10 ml x 2). The combined organic layers were washed with brine (10 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a residue, which was purified by flash chromatography on silica gel (eluting with 3%-30% ethyl acetate in hexane) to give tert-butyl 4 -(7-bromo-2,6-dichloro-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate (520 mg, 90%) as a yellow solid. LC/MS (ESI) m/z: 479.0/481.0 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.50 (s, 9H), 3.69-3.62 (m, 4H), 3.94-3.85 (m, 4H), 7.77 (d, J=2.0 Hz, 1H).

[00498] Step 5: Preparation of tert-butyl 4-(7-bromo-6-chloro-2-((3-ethoxy-3-oxopropyl)amino)-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate

[00499] To a solution of tert-butyl 4-(7-bromo-2,6-dichloro-8-fluoroquinazolin-4-yl)piperazin-1-carboxylate (100 mg, 0.2 mmol) in isopropanol (3 ml) was added ethyl 3-aminopropanoate hydrochloride (91.8 mg, 0.6 mmol) and N-ethyl-N-isopropylpropan-2-amine (78 mg, 0.6 mmol), and refluxed for 20 hours. The reaction mixture was partitioned between ethyl acetate (20 ml) and water (10 ml). The organic layer was collected, and the aqueous layer was extracted with ethyl acetate (15 ml x 2). The combined organic layers were washed with brine (10 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a residue, which was purified by flash chromatography on silica gel (eluting with 10%-50% ethyl acetate in hexane) to give tert-butyl- 4-(7-bromo-6-chloro-2-((3-ethoxy-3-oxopropyl)amino)-8-fluoroquinazolin-4-yl)piperazin-1-carboxylate (95 mg, 81%) as colorless oil . ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.27 (t, J=7.2 Hz, 3H), 1.49 (s, 9H), 2.60-2.70 (m, 2H), 3 .55-3.70 (m, 8H), 3.84-3.75 (m, 2H), 4.16 (q, J=7.2 Hz, 2H), 5.67 (br, 1H), 7.57 (d, J=2.0 Hz, 1H).

[00500] Step 6: Preparation of tert-butyl-4-(6-chloro-2-((3-ethoxy-3-oxopropyl)amino)-8-fluoro-7-(3-(methoxymethoxy)naphthalene-1-yl )quinazolin-4-yl)piperazine-1-carboxylate

[00501] To a solution of tert-butyl-4-(7-bromo-6-chloro-2-((3-ethoxy-3-oxopropyl)amino)-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate (820 mg, 1.46 mmol) in 1,4-dioxane (16.5 ml) and water (4 ml) was added 2-(3-(methoxymethoxy)naphthalene-1-yl)-4,4,5,5-tetramethyl -1,3,2-dioxaborolane (550 mg, 1.75 mmol), sodium carbonate (387 mg, 3.65 mmol) and 1,G-bis(diphenylphosphino)ferrocenepalladium(II)dichloride (110 mg, 0.15 mmol) at room temperature in a nitrogen atmosphere; the mixture was degassed with nitrogen three times. The resulting mixture was heated at reflux for 2 hours. The reaction mixture was partitioned between ethyl acetate (30 ml) and water (20 ml); the organic layer was collected, and the aqueous layer was extracted with ethyl acetate (20 ml×2). The combined organic layers were washed with brine (40 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by flash chromatography on silica gel (eluting with 10-33% ethyl acetate in hexane) to give tert-butyl- 4-(6-chloro-2-((3-ethoxy-3-oxopropyl)amino)-8-fluoro-7-(3-(methoxymethoxy)naphthalene-1-yl)quinazolin-4-yl)piperazin-1- carboxylate (940 mg, 78%) as a white foam. LC/MS (ESI) m/z: 668.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.26 (t, J=7.2 Hz, 3H), 1.51 (s, 9H), 2.67 (t, J=6.0 Hz, 2H), 3.55 (s, 3H), 3.62-3.75 (m, 8H), 3.79-3.83 (m, 2H), 4.16 (q, J=7.2 Hz , 2H), 5.38-5.29 (m, 2H), 5.57-5.65 (m, 1H), 7.19 (d, J=2.4 Hz, 1H), 7.30- 7.26 (m, 1H), 7.37 (d, J=8.4 Hz, 1H), 7.49-7.42 (m, 1H), 7.52 (d, J=2.4 Hz , 1H), 7.66 (d, J=1.4 Hz, 1H), 7.82 (d, J=8.2 Hz, 1H).

[00502] Step 7: Preparation of ethyl-3-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl) amino) propanoate hydrochloride

[00503] To a solution of tert-butyl-4-(6-chloro-2-((3-ethoxy-3-oxopropyl)amino)-8-fluoro-7-(3-(methoxymethoxy)naphthalene-1-yl)quinazoline -4-yl piperazin-1-carboxylate (940 mg, 1.4 mmol) in ethanol (10 ml) was added hydrogen chloride in dioxane (10 ml, 4 M) at room temperature and stirred overnight.Volatile components were evaporated at reduced pressure to obtain ethyl 3-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl)amino)propanoate hydrochloride (1 .4 mmol) LC/MS (ESI) m/z: 524.4 [M+1] ⁺ .

[00504] Step 8: Preparation of ethyl 3-((7-(3-acetoxynaphthalin-1-yl)-4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoroquinazolin-2-yl)amino )propanoate

[00505] To a suspension of ethyl 3-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl)amino)propanoate hydrochloride ( 1.4 mmol) in dichloromethane (30 ml), triethylamine (850 mg, 8.4 mmol) and acetyl chloride (328 mg, 4.2 mmol) were added at room temperature and stirred for 1 hour. The reaction mixture was concentrated in vacuo to give a residue, which was purified on a silica gel column (eluting with 2% methanol in dichloromethane) to give ethyl 3-((7-(3-acetoxynaphthalene-1-yl)-4-(4-acetylpiperazine-1 -yl)-6-chloro-8-fluoroquinazolin-2-yl)amino)propanoate (735 mg, two-step yield 86%) as a light yellow oil. LC/MS (ESI) m/z: 608.7 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.26 (t, J=7.2 Hz, 3H), 2.18 (s, 3H), 2.36 (s, 3H), 2.67 ( t, J=6.0 Hz, 2H), 3.66-3.88 (m, 10H), 4.17 (q, J=7.2 Hz, 2H), 5.64 (t, J=5 ,2 Hz, 1H), 7.23 (d, J=2.4 Hz, 1H), 7.43-7.36 (m, 1H), 7.45 (d, J=8.2 Hz, 1H ), 7.49-7.53 (m, 1H), 7.66 (d, J=1.6 Hz, 1H), 7.73 (d, J=2.2 Hz, 1H), 7.89 (d, J=8.2 Hz, 1H).

[00506] Step 9: Preparation of 3-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino) propanoic acid

[00507] To a solution of ethyl 3-((7-(3-acetoxynaphthalin-1-yl)-4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoroquinazolin-2-yl)amino)propanoate (735 mg, 1.2 mmol) in tetrahydrofuran (6 ml)-water (2 ml) was added lithium hydroxide monohydrate (241 mg, 6.0 mmol) at room temperature and stirred for 2 hours. The mixture solution was cooled to room temperature, acidified with dilute hydrochloric acid (3 N) to pH 3-4, and extracted with dichloromethane (10 ml x 2). The organic layers were combined, washed with brine (10 ml), dried over sodium sulfate and concentrated under reduced pressure to give 3-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7)- (3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)propanoic acid as a white solid (470 mg, 78%). LC/MS (ESI) m/z: 538.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 2.07 (s, 3H), 2.55-2.65 (m, 2H), 3.45-3.85 (m, 8H), 4 .19-3.88 (m, 2H), 7.07 (s, 1H), 7.17-7.35 (m, 3H), 7.53-7.38 (m, 1H), 7.70 -8.10 (m, 2H), 10.22-9.94 (m, 1H), 12.33 (br, 1H).

[00508] Example of the synthesis of 3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)propanoic acid

[00509] Step 1: Preparation of ethyl 3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino )propanoate

[00510] To a solution of ethyl 3-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl)amino)propanoate hydrochloride ( 630 mg, 1.12 mmol) and triethylamine (341 mg, 3.36 mmol) in dichloromethane (5 ml), acryloyl chloride (166 mg, 1.68 mmol) was added at 0°C, stirred at room temperature for 1 hour . The mixture was quenched with water (20 ml) at 0°C and extracted with dichloromethane (20 ml). The organic layer was collected, washed with brine (20 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by flash chromatography on silica gel (eluting with 2-5% methanol in dichloromethane) to give ethyl 3- ((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)propanoate (425 mg, 66%) in in the form of an off-white solid.

[00511] Step 2: Preparation of 3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino) propanoic acid

[00512] Mixture of ethyl 3-((4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)propanoate 3 -((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)propanoate (425 mg, 0.75 mmol) and lithium hydroxide monohydrate (63 mg, 1.5 mmol) in tetrahydrofuran (4 ml)-water (1 ml)-methanol (1 ml) were stirred at room temperature for 1 hour. The reaction mixture was acidified with dilute hydrochloric acid (1 N) to pH 6-7 and extracted with dichloromethane (10 ml (3). The combined organic layers were dried over sodium sulfate and concentrated under reduced pressure to give 3-((4-(4-carbamoylpiperazine -1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)propanoic acid (400 mg, 99%) as a light yellow solid. LC/MS (ESI) m/z: 550.1 [M+1] ⁺ .

[00513] Synthesis example of 2,2,5-trimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl 4-methylbenzenesulfonate

[00514] Step 1: Preparation of 2-(2-(2-hydroxyethoxy)ethoxy)ethyl-4-methylbenzenesulfonate

[00515] To a mixture of 2-[2-(2-hydroxyethoxy)ethoxy]ethanol (5.00 g, 33.29 mmol, 1.00 eq.) and 4-methylbenzenesulfonyl chloride (1.59 g, 8.32 mmol, 0.25 eq.) in dichloromethane (50 mL), triethylamine (1.68 g, 16.65 mmol, 2.3 mL, 0.5 eq.) was added in one portion at 25° C. under nitrogen. The mixture was stirred at 25°C for 12 hours. The mixture was washed with brine (30 ml x 2), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel (petroleum ether/ethyl acetate 1/1 to 0:1) to give 2-[2-(2-hydroxyethoxy)ethoxy]ethyl 4-methylbenzenesulfonate (1.72 g, 5.65 mmol , yield 17%) in the form of a colorless oil. LC/MS (ESI) m/z: 327.0 [M+23] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.81 (d, J=8.3 Hz, 2H), 7.35 (d, J=8.0 Hz, 2H), 4.22-4, 15 (m, 2H), 3.79-3.67 (m, 4H), 3.67-3.56 (m, 6H), 2.46 (s, 3H).

[00516] Step 2: Preparation of 2-(2-(2-(methylamino)ethoxy)ethoxy)ethan-1-ol

[00517] Methamine (41.27 g, 398.65 mmol, 50 mL, 30% in ethanol, 24.27 eq.) was added to 2-[2-(2-hydroxyethoxy)ethoxy]ethyl 4-methylbenzenesulfonate (5 g , 16.43 mmol, 1 eq.). The mixture was stirred at 80°C for 12 hours. The mixture was cooled to 20°C and concentrated in vacuo. 2-[2-[2-(methylamino)ethoxy]ethoxy]ethanol (2.68 g, 16.43 mmol) was obtained as a colorless oil.

[00518] Step 3: Preparation of tert-butyl (2-(2-(2-hydroxyethoxy)ethoxy)ethyl)(methyl)carbamate.

[00519] To a solution of 2-[2-[2-(methylamino)ethoxy]ethoxy]ethanol (2.68 g, 16.43 mmol, 1 eq.) and di-tert-butyl dicarbonate (4.30 g, 19. 70 mmol, 4.5 mL, 1.2 eq.) in dichloromethane (20 mL), triethylamine (3.32 g, 32.84 mmol, 4.5 mL, 2 eq.) was added. The mixture was stirred at 20°C for 12 hours. The mixture was quenched by adding water (20 ml) and the aqueous phase was extracted with ethyl acetate (20 ml x 3). The combined organic phase was washed with brine (20 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel (petroleum ether/ethyl acetate 3/1 to 1/3) to give tert-butyl-N-[2-[2-(2-hydroxyethoxy)ethoxy]ethyl]-N-methyl- carbamate (3.2 g, 12.15 mmol, 74% yield) as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 3.70-3.80 (m, 2H), 3.54-3.70 (m, 8H), 3.41 (br s, 2H), 2, 92 (s, 3H), 1.46 (s, 9H).

[00520] Step 4: Preparation of 2,2,5-trimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl 4-methylbenzenesulfonate

[00521] To a solution of tert-butyl N-[2-[2-(2-hydroxyethoxy)ethoxy]ethyl]-N-methylcarbamate (600 mg, 2.28 mmol, 1 eq.) and triethylamine (461 mg, 4. 56 mmol, 0.6 mL, 2 eq.) in dichloromethane (5 mL), p-toluenesulfonyl chloride (651 mg, 3.42 mmol, 1.5 eq.) was added. The mixture was stirred at 20°C for 12 hours. The mixture was diluted with water (20 ml). The aqueous phase was extracted with ethyl acetate (20 ml x 3). The combined organic phase was washed with brine (20 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 10/1 to 3/1) to give 2-[2-[2-[tert-butoxycarbonyl(methyl)amino]ethoxy]ethoxy]ethyl 4-methylbenzenesulfonate (750 mg, 1.80 mmol, 79% yield) as a colorless oil. LC/MS (ESI) m/z: 440.1 [M+23] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.81 (d, J=8.0 Hz, 2H), 7.35 (d, J=8.0 Hz, 2H), 4.19-4, 15 (m, 2H), 3.70 (t, J=4.8 Hz, 2H), 3.61-3.51 (m, 6H), 3.38 (br s, 2H), 2.90 ( s, 3H), 2.46 (s, 3H), 1.45 (s, 9H).

[00522] Example of synthesis of 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione

[00523] Step 1: Preparation of 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione

[00524] To a mixture of 3-aminopiperidine-2,6-dione (10.90 g, 66.22 mmol, 1.1 eq., hydrochloride) and 4-fluoroisobenzofuran-1,3-dione (10 g, 60.20 mmol, 1 eq.) in acetic acid (150 mL), potassium acetate (18.32 g, 186.63 mmol, 3.1 eq.) was added in one portion at 100°C under nitrogen. The mixture was stirred at 100°C for 16 hours. The mixture was cooled to 20°C and concentrated under reduced pressure at 55°C. The residue was triturated with ethyl acetate (100 ml) and water (30 ml), the mass was collected under reduced pressure and dried under high vacuum. 2-(2,6-dioxo-3-piperidyl)-4-fluoroisoindoline-1,3-dione (8.9 g, 32.22 mmol, 53% yield) was obtained as a black solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.15 (br s, 1H), 7.95 (dt, J=4.6, 7.9 Hz, 1H), 7.82-7, 70 (m, 2H), 5.17 (dd, J=5.3, 12.9 Hz, 1H), 2.99-2.81 (m, 1H), 2.66-2.52 (m, 2H), 2.13-2.02 (m, 1H).

[00525] Synthesis example of (S)-3-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazoline -2-yl)amino)propanoic acid

[00526] Step 1: Preparation of 3-fluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol.

[00527] Boron tribromide (44 g, 176.52 mmol, 3 .0 eq.) at 0°C. The mixture was then stirred at 0°C for 0.5 hours. The mixture was diluted with ice water (100 ml) at 0°C, extracted with ethyl acetate (200 ml (3), washed with brine (200 ml), dried over anhydrous sodium sulfate, filtered and then concentrated. To the residue in tetrahydrofuran (100 ml) was added 2,3-dimethylbutane-2,3-diol (13.91 g, 117.68 mmol, 2 eq.) The mixture was then stirred at 20° C. for 12 hours. The mixture was then concentrated. The residue was purified by column chromatography on silicon dioxide (petroleum ether) to give 3-fluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan 2-yl)phenol (6 g, 25.20 mmol, 42% yield) as a white solid ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.37 (s, 1H), 7.36-7.28 (m, 1H), 6.69 (d, J=8, 4 Hz, 1H), 6.57 (t,.7=8.8 Hz, 1H), 1.40 (s, 12H).

[00528] Step 2: Preparation of tert-butyl-4-(7-bromo-6-chloro-8-fluoro-2-((3-methoxy-3-oxopropyl)amino)quinazolin-4-yl)piperazin-1- carboxylate

[00529] To a mixture of methyl 3-aminopropanoate (16.57 g, 118.71 mmol, 3 eq., hydrochloride) and tert-butyl-4-(7-bromo-2,6-dichloro-8-fluoroquinazoline-4 -yl)piperazine-1-carboxylate (19 g, 39.57 mmol, 1 eq.) in isopropanol (600 ml) was added N-diisopropylethylamine (25.57 g, 197.85 mmol, 34.46 ml, 5 eq. ) in one portion at 20°C in a nitrogen atmosphere. The mixture was stirred at 95°C for 12 hours. The mixture was cooled to 20°C and concentrated under reduced pressure at 45°C. The residue was purified by silica gel chromatography (petroleum ether/ethyl acetate 10/1, 3/1) to give tert-butyl-4-[7-bromo-6-chloro-8-fluoro-2-[(3-methoxy- 3-oxo-propyl)amino]quinazolin-4-yl]piperazine-1-carboxylate (10.6 g, 19.38 mmol, 49% yield) as a yellow solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.75 (s, 1H), 7.65-7.32 (m, 1H), 3.64-3.52 (m, 10H), 3 .34 (s, 3H), 2.66 (br d, J=9.2 Hz, 2H), 1.43 (s, 9H).

[00530] Step 3: Preparation of tert-butyl-4-(6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-((3-methoxy-3-oxopropyl)amino)quinazoline- 4-yl)piperazine-1-carboxylate

[00531] To a solution of tert-butyl-4-[7-bromo-6-chloro-8-fluoro-2-[(3-methoxy-3-oxopropyl)amino]quinazolin-4-yl]piperazine-1-carboxylate ( 6.8 g, 12.44 mmol, 1 eq.) and 3-fluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (7.43 g , 31.21 mmol, 2.51 eq.) in toluene (140 ml) was added [2-(2-aminophenyl)phenyl]chloro-palladium; dicyclohexyl-[2-(2,6-dimethoxyphenyl)phenyl]phosphane (448 mg, 0.62 mmol, 0.05 eq.) and potassium phosphate (1.5 M, 24.87 ml, 3 eq.). The reaction mixture was degassed and charged with nitrogen three times and then stirred at 75°C for 30 hours. Ethyl acetate (80 ml) and water (100 ml) were added and the mixture was separated. The organic layer was dried over sodium sulfate and then concentrated in vacuo to obtain a residue. The residue was purified by flash chromatography on silica gel (0-100% ethyl acetate in petroleum ether) to give tert-butyl-4-[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-[ (3-methoxy-3-oxopropyl)amino]quinazolin-4-yl]piperazine-1-carboxylate (4.2 g, 6.90 mmol, 55% yield, 95% purity) as a yellow solid. LC/MS (ESI) m/z: 578.2 [M+1] ⁺ .

[00532] Step 4: Preparation of tert-butyl (S)-4-(6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-((3-methoxy-3-oxopropyl)amino )quinazolin-4-yl)piperazin-1-carboxylate and tert-butyl (R)-4-(6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-((3-methoxy -3-oxopropyl)amino)quinazolin-4-yl)piperazine-1-carboxylate

[00533] Tert-butyl-4-[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-[(3-methoxy-3-oxopropyl)amino]quinazolin-4-yl] piperazine-1-carboxylate (5 g, 8.65 mmol, 1 eq.) was separated by SFC. We obtained tert-butyl (S)-4-(6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-((3-methoxy-3-oxopropyl)amino)quinazolin-4-yl )piperazine-1-carboxylate (2.38 g, 3.85 mmol, 89% yield, 93% purity) as a yellow solid. Tert-butyl (R)-4-(6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-((3-methoxy-3-oxopropyl)amino)quinazolin-4-yl was obtained )piperazine-1-carboxylate (2.5 g, 4.15 mmol, 96% yield, 96% purity) as a yellow solid.

[00534] Step 5: Preparation of tert-butyl (S)-3-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro- 6-Hydroxyphenyl)quinazolin-2-yl)amino)propanoic acid

[00535] To a solution of tert-butyl-4-[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-[(3-methoxy-3-oxo-propyl)amino]quinazoline- 4-yl]piperazine-1-carboxylate (2.38 g, 3.83 mmol, 1 eq.) mixed with tetrahydrofuran (12 ml) and methanol (12 ml) was added water (12 ml) and lithium hydroxide monohydrate (450 mg., 10.73 mmol, 2.80 eq.). The reaction mixture was stirred at 20°C for 16 hours. The pH of the aqueous layer was adjusted to 6 by adding 1 N hydrochloric acid. The resulting mixture was extracted with ethyl acetate (70 ml x 2). The organic layers were dried over sodium sulfate and then concentrated in vacuo to give (S)-3-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-6-chloro-8-fluoro-7-(2 -fluoro-6-hydroxyphenyl)quinazolin-2-yl)amino)propanoic acid (2.02 g, 3.58 mmol, 93% yield) as a yellow solid.

[00536] Synthesis example of (R)-3-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazoline -2-yl)amino)propanoic acid

[00537] Step 1: Preparation of (R)-3-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl )quinazolin-2-yl)amino)propanoic acid

[00538] To a solution of tert-butyl-4-[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-[(3-methoxy-3-oxo-propyl)amino]quinazoline- 4-yl]piperazine-1-carboxylate (2.5 g, 4.15 mmol, 1 eq.) mixed with tetrahydrofuran (12 ml) and methanol (12 ml) was added water (12 ml) and lithium hydroxide monohydrate (488 mg, 11.63 mmol, 2.80 eq.). The reaction mixture was stirred at 20°C for 16 hours. The pH of the mixture was adjusted to 6 by adding 1 N hydrochloric acid. The resulting mixture was extracted with ethyl acetate (80 ml x 2). The organic layer was dried over sodium sulfate and then concentrated in vacuo to give (R)-3-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-6-chloro-8-fluoro-7-( 2-Fluoro-6-hydroxyphenyl)quinazolin-2-yl)amino)propanoic acid (2.24 g, 3.97 mmol, 95% yield) as a yellow solid.

[00539] Synthesis example of (S)-3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazolin-2-yl) amino)propanoic acid

[00540] Step 1: Preparation of (S)-3-((6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-(piperazin-1-yl)quinazolin-2-yl) amino)propanoic acid

[00541] To a solution of (S)-3-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazoline -2-yl)amino)propanoic acid (1.0 g, 1.77 mmol, 1 eq.) in dioxane (10 mL) added hydrogen chloride/dioxane (4 M, 30.00 mL, 67.68 eq.) . The mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated under reduced pressure to give the product (S)-3-((6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-(piperazin-1-yl)quinazolin-2- yl)amino)propanoic acid (0.88 g, 1.76 mmol, 99% yield, hydrochloride) as a white solid. LC/MS (ESI) m/z: 464.0 [M+1] ⁺ .

[00542] Step 2: Preparation of (S)-3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazoline-2- yl)amino)propanoic acid

[00543] To a solution of (S)-3-((6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-(piperazin-1-yl)quinazolin-2-yl)amino) propanoic acid (0.88 g, 1.76 mmol, 1 eq., hydrochloride) and acryloyl chloride (159 mg, 1.76 mmol, 143.24 μl, saturated sodium bicarbonate (20 ml) in tetrahydrofuran (20 ml), 9.98e-1 eq.). The mixture was stirred at 0°C for 0.5 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent to obtain a residue. The residue was purified by preparative HPLC. The compound (S)-3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazolin-2-yl)amino)propane was obtained acid (0.52 g, 1.00 mmol, 57% yield) as a white solid. ¹ H-NMR (400 MHz, CD ₃ OD) δ 7.77 (s, 1H), 7.33-7.25 (m, 1H), 6.86-6.66 (m, 3H), 6, 26 (dd, J=2.0, 16.8 Hz, 1H), 5.80 (dd, J=2.0, 10.8 Hz, 1H), 3.88 (br s, 8H), 3, 74 (br t, J=6.4 Hz, 2H), 2.63 (t, J=6.4 Hz, 2H).

[00544] Synthesis example of (S)-3-((4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazolin-2-yl) amino)propanoic acid

[00545] Step 1: Preparation of methyl (S)-3-((6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-(piperazin-1-yl)quinazolin-2-yl )amino)propanoate hydrochloride

[00546] To a solution of 3-[[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazolin-2-yl]amino] propanoic acid (1.5 g, 2.66 mmol, 1 eq.) in methanol (5 ml) was added hydrochloric acid in methanol (4 M, 10 ml). The mixture was stirred at 50°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the compound methyl 3-[[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-piperazin-1-ylquinazolin-2-yl]amino]propanoate (1.35 g, hydrochloride) as a yellow solid. LC/MS (ESI) m/z: 478.2 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.63 (brs, 1H), 9.92 (brs, 2H), 8.77-8.43 (m, 1H), 8.08 (s , 1H), 7.38 (q, J=8.0 Hz, 1H), 6.94 (d, J=8.4 Hz, 1H), 6.87-6.72 (m, 1H), 4 ,42-4.15 (m, 4H), 3.86-3.51 (m, 5H), 3.36-3.26 (m, 4H), 2.71-2.55 (m, 2H) .

[00547] Step 2: Preparation of methyl (S)-3-((4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazoline-2 -yl)amino)propanoate

[00548] To a solution of methyl 3-[[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]propanoate (1 .35 g, 2.62 mmol, 1 eq., hydrochloride) and triethylamine (266 mg, 2.62 mmol, 1 eq.) in N,N-dimethylformamide (10 ml) and tetrahydrofuran (5 ml), isocyanato( trimethyl)silane (302 mg, 2.62 mmol, 1 eq.). The mixture was stirred at 20°C for 0.5 hour. The reaction mixture was concentrated under reduced pressure to obtain the compound methyl 3-[[4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazoline-2- yl]amino]propanoate (1.37 g) as a yellow oil. LC/MS (ESI) m/z: 521.2 [M+1] ⁺ .

[00549] Step 3: Preparation of (S)-3-((4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazoline-2- yl)amino)propanoic acid

[00550] To a solution of methyl 3-[[4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazolin-2-yl]amino]propanoate ( Lithium hydroxide monohydrate (276 mg, 6.57 mmol, 2.5 eq.) was added to tetrahydrofuran (10 mL) and water (3 mL). The mixture was stirred at 20°C for 0.5 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was diluted with water (10 ml), acidified to pH=3 with hydrochloric acid (1 M), and some precipitate formed when hydrochloric acid was added. The resulting mixture was filtered and the filter cake was evaporated to dryness under reduced pressure to obtain the compound 3-[[4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl )quinazolin-2-yl]amino]propanoic acid (1.2 g, 2.37 mmol, 90% yield) as a yellow solid. LC/MS (ESI) m/z: 507.2 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 12.41 (brs, 1H), 10.57-10.08 (m, 1H), 8.19-7.94 (m, 1H), 7 ,93-7.64 (m, 1H), 7.44-7.26 (m, 1H), 6.98-6.73 (m, 2H), 6.11 (brs, 2H), 4.19 -3.91 (m, 2H), 3.77-3.46 (m, 4H), 3.43-3.36 (m, 4H), 2.58 (t, J=6.4 Hz, 2H ).

[00551] Example of the synthesis of 4-(6-chloro-8-fluoro-4-(piperazin-1-yl)quinazolin-7-yl)naphthalene-2-ol hydrochloride

[00552] Step 1: Preparation of 7-bromo-6-chloro-8-fluoroquinazolin-4(3H)-one

[00553] A mixture of 2-amino-4-bromo-5-chloro-3-fluorobenzoic acid (1.5 g, 5.58 mmol) and formimidamide acetate (2.3 g, 22.35 mmol) in ethanol (30 ml ) was stirred at reflux for 24 hours. The mixture was concentrated and the residue was partitioned between water (20 ml) and ethyl acetate (20 ml). The organic layer was washed with brine (10 ml), dried over sodium sulfate and concentrated under reduced pressure to give 7-bromo-6-chloro-8-fluoroquinazolin-4(3H)-one (1.2 g, 77%) as light yellow solid.

[00554] Step 2: Preparation of 7-bromo-4,6-dichloro-8-fluoroquinazoline

[00555] To a solution of 7-bromo-6-chloro-8-fluoroquinazolin-4(3H)-one (1.2 g, 4.3 mmol) in sulfur dioxide (16 ml) was added N,N-dimethylformamide (1 ml ) at room temperature. The mixture was stirred at reflux for 16 hours. The mixture was concentrated and the residue was dissolved in dichloromethane (20 ml). The organic layer was poured into ice water (20 ml), washed with aqueous sodium bicarbonate (1 N, 10 ml) and brine (10 ml), dried over sodium sulfate and concentrated under reduced pressure to obtain a crude residue, which was purified by flash -silica gel chromatography (eluting with 5% ethyl acetate in hexane) to give 7-bromo-4,6-dichloro-8-fluoroquinazoline (1.07 g, 83%) as a yellow solid.

[00556] Step 3: Preparation of tert-butyl 4-(7-bromo-6-chloro-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate

[00557] A mixture of 7-bromo-4,6-dichloro-8-fluoroquinazoline (1.07 g, 3.61 mmol), tert-butylpiperazine-1-carboxylate (674 mg, 3.61 mmol) and triethylamine (1, 1 g, 10.85 mmol) in 1,4-dioxane (11 ml) was stirred at 50°C for 30 minutes. The reaction mixture was concentrated and the residue was partitioned between ethyl acetate (15 ml) and water (20 ml). The organic layer was collected, washed with brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by silica gel flash chromatography (eluting with 20-25% ethyl acetate in hexane) to give tert-butyl -4-(7-bromo-6-chloro-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate (1 g, 62%) as a white solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.50 (s, 9H), 3.65, 3.80 (two singles, 8H), 7.79 (s, 1H), 8.77 (s, 1H).

[00558] Step 4: Preparation of tert-butyl-4-(6-chloro-8-fluoro-7-(3-(methoxymethoxy)naphthalene-1-yl)quinazolin-4-yl)piperazine-1-carboxylate

[00559] To a suspension of tert-butyl 4-(7-bromo-6-chloro-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate (1 g, 2.24 mmol), 2-(3-(methoxymethoxy )naphthalene-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (705 mg, 2.24 mmol) and potassium carbonate (624 mg, 4.48 mmol) in dioxane (8 tetrakis(triphenylphosphine)palladium (0) (520 mg, 0.45 mmol) was added to ml)-water (2 ml) at room temperature under a nitrogen atmosphere; the mixture was degassed and charged again with nitrogen three times. The resulting mixture was refluxed for 12 hours. The mixture was concentrated and the residue was partitioned between water (10 ml) and ethyl acetate (10 ml). The organic layer was collected, washed with brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by silica gel flash chromatography (eluting with 18-33% ethyl acetate in hexane) to give tert-butyl -4-(6-chloro-8-fluoro-7-(3-(methoxymethoxy)naphthalene-1-yl)quinazolin-4-yl)piperazin-1-carboxylate (420 mg, 34%) as a light yellow solid substances. LC/MS (ESI) m/z: 553.20 [M+1] ⁺ .

[00560] Step 5: Preparation of 4-(6-chloro-8-fluoro-4-(piperazin-1-yl)quinazolin-7-yl)naphthalene-2-ol hydrochloride

[00561] To a solution of tert-butyl-4-(6-chloro-8-fluoro-7-(3-(methoxymethoxy)naphthalene-1-yl)quinazolin-4-yl)piperazine-1-carboxylate (200 mg, 0 .36 mmol) in methanol (2 ml), a solution of dioxane hydrochloric acid (2 ml) was added at room temperature. The mixture was stirred at room temperature for 1 hour. The mixture was concentrated and the solid was washed with ethyl acetate (5 ml). The solid was collected and dried to obtain 4-(6-chloro-8-fluoro-4-(piperazin-1-yl)quinazolin-7-yl)naphthalene-2-ol hydrochloride (190 mg). LC/MS (EST) m/z: 409.10 [M+1] ⁺ .

[00562] Synthesis example of (2S,4R)-1-((S)-15-(tert-butyl)-13-oxo-5,8,11-trioxa-2,14-diazahexadecane-16-oil)-4 -hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[00563] Step 1: Preparation of tert-butyl 2,2,5-trimethyl-4-oxo-3,8,11,14-tetraoxa-5-azahexadecane-16-oate.

[00564] A solution of tert-butyl (2-(2-(2-hydroxyethoxy)ethoxy)ethyl)(methyl)carbamate (600 mg, 2.28 mmol), tetrabutylammonium chloride (635 mg, 2.38 mmol), tert- butyl 2-bromoacetate (445 mg, 2.28 mmol) and sodium hydroxide (12 ml, 35% in water) dichloromethane (12 ml) were stirred at room temperature for 4 hours. The organic layer was collected, and the aqueous layer was extracted with dichloromethane (10 ml x 2). The combined organic layers were washed with brine (20 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a residue, which was purified by flash chromatography on silica gel (eluting with 10-20% ethyl acetate in cyclohexane) to give tert-butyl-2 ,2,5-trimethyl-4-oxo-3,8,11,14-tetraoxa-5-azahexadecane-16-oate (300 mg, 35%) as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.45,1.47 (two singles, 18H), 2.91 (s, 3H), 3.32-3.46 (m, 2H), 3, 58-3.73 (m, 10H), 4.02 (s, 2H).

[00565] Step 2: Preparation of 2,2,5-trimethyl-4-oxo-3,8,11,14-tetraoxa-5-azahexadecane-16-oic acid.

[00566] Mixture of tert-butyl 2,2,5-trimethyl-4-oxo-3,8,11,14-tetraoxa-5-azahexadecane-16-oate (100 mg, 0.26 mmol) and lithium hydroxide monohydrate (23 mg, 0.53 mmol) in tetrahydrofuran (1 ml)-water (0.5 ml)-methanol (0.5 ml) was stirred at room temperature for 1 hour. The mixture was concentrated and the residue was acidified with aqueous hydrochloric acid (1 N) to pH 4-5 and extracted with dichloromethane (10 ml). The organic layer was washed with brine (5 ml), dried over sodium sulfate and concentrated under reduced pressure to give 2,2,5-trimethyl-4-oxo-3,8,11,14-tetraoxa-5-azahexadecane-16-oine acid (77 mg, 90%) as a colorless oil.

[00567] Step 3: Preparation of tert-butyl ((S)-13-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl) phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-14,14-dimethyl-11-oxo-3,6,9-trioxa-12-azapentadecyl)(methyl)carbamate

[00568] To a solution of 2,2,5-trimethyl-4-oxo-3,8,11,14-tetraoxa-5-azahexadecane-16-oic acid (77 mg, 0.24 mmol), (2S,4R) -1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine 2 HATU (2-( 7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluron hexafluorophosphate) (182 mg, 0.48 mmol) at 0°C, the resulting mixture was allowed to warm to room temperature and stirred at room temperature within 10 minutes. The mixture was partitioned between ethyl acetate (20 ml) and water (20 ml). The organic layer was collected, washed with brine (20 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by silica gel flash column chromatography (eluting with 5% methanol in dichloromethane) to give tert-butyl ( (S)-13-((2S,4R)-4-hydroxy-2-(((S-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl) -14,14-dimethyl-11-oxo-3,6,9-trioxa-12-azapentadecyl)(methyl)carbamate (140 mg, 78%) as a light yellow oil. LC/MS (ESI) m/z : 748.40 [M+1] ⁺ .

[00569] Step 4: Preparation of (2S,4R)-1-((S)-15-(tert-butyl)-13-oxo-5,8,11-trioxa-2,14-diazahexadecane-16-oil) -4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[00570] To a solution of tert-butyl ((S)-13-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl) ethyl)carbamoyl)pyrrolidine-1-carbonyl)-14,14-dimethyl-11-oxo-3,6,9-trioxa-12-azapentadecyl)(methyl)carbamate (70 mg, 0.093 mmol) in dichloromethane (1 ml) 2,2,2-trifluoroacetic acid (0.5 ml) was added at room temperature. The mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure to give (2S,4R)-1-((S)-15-(tert-butyl)-13-oxo-5,8,11-trioxa-2,14-diazahexadecane-16-oil) -4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (70 mg) as a light yellow oil.

[00571] Synthesis example of (2S,4R)-1-((S)-18-(tert-butyl)-16-oxo-5,8,11,14-tetraoxa-2,17-diazanonadecan-19-oil) -4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[00572] Step 1: Preparation of 2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl-4-methylbenzenesulfonate

[00573] To a mixture of 2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethanol (5 g, 25.74 mmol, 4.4 mL, 1 eq.) and 4-methylbenzenesulfonyl chloride (1.23 g , 6.44 mmol, 0.25 eq.) in dichloromethane (50 ml.), triethylamine (1.30 g, 12.87 mmol, 1.8 ml, 0.5 eq.) was added in one portion at 25°C in nitrogen atmosphere. The mixture was stirred at 25°C for 12 hours. The mixture was washed with brine (30 ml x 2), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel (petroleum ether/ethyl acetate 1/1 to 0/1) to give 2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethyl 4-methylbenzenesulfonate (1.98 g , 5.68 mmol, yield 22%) as a colorless oil. LC/MS (ESI) m/z: 371.0 [M+23] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.82 (d, J=8.3 Hz, 2H), 7.36 (d, J=8.2 Hz, 2H), 4.21-4, 11 (m, 2H), 3.75-3.61 (m, 14H), 2.47 (s, 3H).

[00574] Step 2: Preparation of 5,8,11-trioxa-2-azatridecan-13-ol.

[00575] A solution of 2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethyl-4-methylbenzenesulfonate (6 g, 17.22 mmol, 1 eq.) in methamine (16.21 g, 172.21 mmol, purity 33%, 10 eq.) was stirred at 80°C for 16 hours. The reaction mixture was concentrated in vacuo to obtain a residue. 2-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]ethanol (4.6 g) was obtained as a light yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.77 (d, J=8.2 Hz, 2H), 7.17 (d, J=7.9 Hz, 2H), 3.84-3, 71 (m, 4H), 3.69-3.60 (m, 10H), 3.21-3.09 (m, 2H), 2.82-2.63 (m, 3H).

[00576] Step 3: Preparation of tert-butyl (2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl)(methyl)carbamate

[00577] To a solution of 2-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]ethanol (3.86 g, 18.62 mmol, 1 eq.) in dichloromethane (30 mL) was added di- tert-butyl dicarbonate (4.88 g, 22.35 mmol, 5.13 ml, 1.2 eq.) at 0°C, then stirred at 15°C for 16 hours. Triethylamine (3.77 g, 37.25 mmol, 5.18 mL, 2 eq.) was added and then stirred at 15°C for 12 hours. The reaction mixture was concentrated in vacuo to obtain a residue. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1 to 1/3) to give tert-butyl-N-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethyl] -N-methyl-carbamate (5.74 g) as a light yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 3.75-3.47 (m, 14H), 3.38 (br s, 2H), 2.90 (s, 3H), 1.44 (s, 9H).

[00578] Step 4: Preparation of ethyl 2,2,5-trimethyl-4-oxo-3,8,11,14,17-pentaoxa-5-azanonedecane-19-oate

[00579] To a mixture of tert-butyl-N-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethyl]-N-methylcarbamate (1 g, 3.25 mmol, 1 eq.) and diacetoxyrhodium (143 mg, 0.32 mmol, 0.1 eq.) in dichloromethane (20 ml) was added ethyl 2-diazoacetate (2.47 g, 19.50 mmol, 2.27 ml, 90% purity, 6 eq.) three portions drop by drop at a temperature from minus 5 to 0°C. The reaction mixture was then stirred at 10°C for 16 hours. Acetic acid (0.4 ml) was added to the reaction mixture. The reaction mixture was then extracted with dichloromethane (30 ml x 3) and concentrated in vacuo to give a residue. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 20/1 to 1/3) to give ethyl 2-[2-[2-[2-[2-[tert-butoxycarbonyl(methyl)amino]ethoxy ]ethoxy]ethoxy]ethoxy]acetate (600 mg, 1.52 mmol, 46% yield) as green oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.21 (q, J=7.2 Hz, 2H), 4.15 (s, 2H), 3.75-3.67 (m, 4H), 3.66 (s, 4H), 3.64-3.51 (m, 6H), 3.39 (br s, 2H), 2.91 (s, 3H), 1.50-1.39 (m , 9H), 1.31-1.26 (m, 3H).

[00580] Step 5: Preparation of 2,2,5-trimethyl-4-oxo-3,8,11,14,17-pentaoxa-5-azanonedecane-19-oic acid

[00581] To a solution of ethyl 2-[2-[2-[2-[2-[tert-butoxycarbonyl(methyl)amino]ethoxy]ethoxy]ethoxy]ethoxy]acetate (600 mg, 1.52 mmol, 1 eq. ) in tetrahydrofuran (5 ml), water (5 ml) and methanol (5 ml), lithium hydroxide monohydrate (365 mg, 15.25 mmol, 10 eq.) was added, then the mixture was stirred at 15°C for 4 hours. 1 M aqueous hydrogen chloride solution was added to the mixture until pH 3.0-4.0, then extracted with dichloromethane (30 ml x 3), then the combined organic layers were washed with brine (10 ml), dried with anhydrous sodium sulfate, filtered and concentrated at reduced pressure. 2-[2-[2-[2-[2-[tert-butoxycarbonyl(methyl)amino]ethoxy]ethoxy]ethoxy]ethoxy]acetic acid (473 mg, 1.29 mmol, 84%)) was obtained as colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.16 (s, 2H), 3.80-3.74 (m, 2H), 3.73-3.57 (m, 12H), 3.41 (br s, 2H), 2.92 (s, 3H), 1.46 (s, 9H).

[00582] Step 6: Preparation of tert-butyl ((S)-16-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl) phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-17,17-dimethyl-14-oxo-3,6,9,12-tetraoxa-15-azaoctadecyl)(methyl)carbamate

[00583] To a solution of 2-[2-[2-[2-[2-[tert-butoxycarbonyl(methyl)amino]ethoxy]ethoxy]ethoxy]ethoxy]acetic acid (250 mg, 0.68 mmol, 1 eq. ), (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl )phenyl]ethyl]pyrrolidine-2-carboxamide (304 mg, 0.68 mmol, 1 eq.) and N,N-diisopropylethylamine (265 mg, 2.05 mmol, 0.36 ml, 3 eq.) in N, N-dimethylformamide (5 ml) was added with hydroxybenzotriazole (111 mg, 0.82 µmol, 1.2 equiv.) and 3-(ethyliminomethyleneamino)-N,N-dimethyl-propan-1-amine hydrochloride (157 mg, 0.82 mmol, 1.2 eq.) successively at 0°C, then stirred at 15°C for 12 hours. The reaction mixture was poured into water (30 ml) and then extracted with ethyl acetate (20 ml x 3). The combined organic phase was washed with brine (10 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative thin-layer chromatography (dichloromethane/methanol 10/1) to give tert-butyl N-[2-[2-[2-[2-[2-[[(1S )-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]- 2,2-dimethylpropyl]amino]-2-oxoethoxy]ethoxy]ethoxy]ethoxy]ethyl]-N-methylcarbamate (405 mg, 0.51 mmol, 74% yield) as a colorless oil. ¹ H-NMR (400 MHz, DMCO-d ₆ ) δ 8.98 (s, 1H), 8.43 (d, J=7.7 Hz, 1H), 7.46-7.41 (m, 2H ), 7.37 (d, J=8.2 Hz, 3H), 5.12 (d, J=3.5 Hz, 1H), 4.91 (q, J=6.9 Hz, 1H), 4.54 (d, J=9.7 Hz, 1H), 4.44 (t, J=8.2 Hz, 1H), 4.28 (br s, 1H), 3.63-3.47 ( m, 15H), 3.30-3.26 (m, 2H), 3.17 (s, 3H), 2.80 (br d, J=4.4 Hz, 3H), 2.45 (s, 3H), 1.77 (ddd, J=4.5, 8.5, 12.9 Hz, 1H), 1.43-1.35 (m, 12H), 0.94 (s, 9H).

[00584] Step 7: Preparation of tert-butyl (2S,4R)-1-((S)-18-(tert-butyl)-16-oxo-5,8,11,14-tetraoxa-2,17-diazanonedecane -19-oil)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[00585] To a solution of tert-butyl N-[2-[2-[2-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S )-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl]amino]-2-oxoethoxy]ethoxy]ethoxy]ethoxy]ethyl] -N-methylcarbamate (120 mg, 0.15 mmol, 1 eq.) in dichloromethane (4 ml) was added hydrochloride/dioxane (4 M, 4 ml, 105.60 eq.), then stirred at 15°C for 2 hours. The reaction mixture was concentrated in vacuo to give (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-[2-[2-[2-(methylamino)ethoxy]ethoxy ]ethoxy]ethoxy]acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (120 mg, hydrochloride) in the form of a colorless oil.

[00586] Synthesis example of (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine -2-carboxamide hydrochloride

[00587] Step 1: Preparation of 4-(4-methylthiazol-5-yl)benzonitrile.

[00588] In a 1 L round bottom flask, purged and maintained under an inert atmosphere of nitrogen, was placed a solution of 4-bromobenzonitrile (20 g, 109.88 mmol, 1.00 eq.) in DMA (250 mL), 4-methyl-1 ,3-thiazole (21.88 g, 220.67 mmol, 2.00 eq.), Pd (OAc) ₂ (743 mg, 3.31 mmol, 0.03 eq.) and KOAc (21.66 g, 220.71 mmol, 2.00 eq.). The resulting solution was stirred for 5 hours at a temperature of 150°C. The reaction mixture was cooled in a water/ice bath and diluted with 1 L of water. The resulting solution was extracted with 3×300 ml of ethyl acetate. The combined organic layers were washed with 3 x 300 mL water and 1 x 300 mL brine, then dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by combined flash chromatography with ethyl acetate/petroleum ether (1:100-1:5). The result was 20 g (91%) of 4-(4-methyl-1,3-thiazol-5-yl)benzonitrile as a beige solid.

[00589] Step 2: Preparation of (4-(4-methylthiazol-5-yl)phenyl)methanamine.

[00590] Place 4-(4-methyl-1,3-thiazol-5-yl)benzonitrile (35 g, 174.77 g) mmol into a 3-L, 3-neck round bottom flask, purged and maintained under an inert atmosphere of nitrogen. , 1.00 eq.) in tetrahydrofuran (1000 ml). LiAlH ₄ (20 g, 526.32 mmol, 3.00 eq.) was then added in portions at 0° C. over 10 minutes. The resulting solution was stirred for 3 hours at 60°C in an oil bath. The reaction was cooled to 0°C in a water/ice bath, then quenched by adding 20 ml water, 20 ml NaOH (15%) and 60 ml water. The resulting solution was extracted with 200 ml of ethyl acetate. Solids were filtered off. The filtrate was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was applied to a silica gel column with a dichloromethane/methanol mixture (10:1). The result was 20 g (56%) of [4-(4-methyl-1,3-thiazol-5-yl)phenyl]methanamine as a yellow oil

[00591] Step 3: Preparation of tert-butyl (2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate

[00592] Place (2S,4R)-1-[(tert-butoxy)carbonyl]-4-hydroxypyrrolidine-2-carboxylic acid (2.7 g, 11.68 mmol, 1.20 eq) into a 50 mL round bottom flask .) in N,N-dimethylformamide (30 ml), DIEA (2.52 g, 19.50 mmol, 1.20 eq.), HATU (4.47 g, 11.76 mmol, 1.20 eq.) , [4-(4-methyl-1,3-thiazol-5-yl)phenyl]methanamine (2 g, 9.79 mmol, 1.00 eq.). The resulting solution was stirred overnight at 25°C. The reaction was then quenched by adding 20 ml water and extracted with 3×20 ml ethyl acetate. The organic layers were combined, dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was applied to a silica gel column with a mixture of dichloromethane/methanol (20:1). The result was 1 g (24%) of tert-butyl (2S,4R)-4-hydroxy-2-([[4-(4-methylthiazol-5-yl)phenyl]methyl]carbamoyl)pyrrolidine-1-carboxylate in as a yellow solid.

[00593] Step 4: Preparation of (2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride.

[00594] Place tert-butyl (2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]carbamoyl into a 1000 mL round bottom flask )pyrrolidine-1-carboxylate (45 g, 107.78 mmol, 1.00 eq.), hydrogen chloride solution (13.44 L) in dioxane (300 ml). The resulting solution was stirred for 2 hours at a temperature of 20°C. Solids were collected by filtration. The result was 37.3 g (98%) of (2S,4R)-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidin-2- carboxamide hydrochloride as a yellow solid.

[00595] Step 5: Preparation of tert-butyl ((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine)1 -yl)-3,3-dimethyl-1-oxobutan-2-yl)carbamate

[00596] Place (2S)-2-[[(tert-butoxy)carbonyl]amino]-3,3-dimethylbutanoic acid (15.73 g, 68.01 mmol, 1.20 eq.) into a 1000 mL round bottom flask. ) in N,N-dimethylformamide (500 ml), DIEA (29.2 g, 225.94 mmol, 4.00 eq), HATU (25.9 g, 68.12 mmol, 1.20 eq) and (2S,4R)-2-amino-5-chloro-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pentanamide (20 g, 56, 52 mmol, 1.00 eq.). The resulting solution was stirred for 16 hours at a temperature of 20°C. The reaction was then quenched by adding 200 ml of water and extracted with 3×100 ml of ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (2:1). The result was 15.2 g (51%) tert-butyl-N-[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3 -thiazol-5-yl)phenyl]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate as a yellow solid.

[00597] Step 6: Preparation of (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl )pyrrolidine-2-carboxamide hydrochloride

[00598] Place tert-butyl N-[(2S)-1-[(2S,4R)-4-hydroxy-2-([[4-(4-methyl-1,3-thiazole) into a 500 mL round bottom flask -5-yl)phenyl]methyl]carbamoyl)pyrrolidin-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate (12 g, 22.61 mmol, 1.00 eq.) in dioxane (20 ml) and a solution of hydrogen chloride (3.584 l) in dioxane (80 ml). The resulting solution was stirred for 2 hours at a temperature of 25°C. Solids were collected by filtration. The result was 5.1 g (48%) of (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl- 1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide hydrochloride as a yellow solid. LC/MS (ESI) m/z: 431 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 9.84-9.82 (s, 1H), 7.58-7.54 (m, 4H), 4.71-4.41 (m, 4H ), 4.13-4.08 (m, 1H), 3.86-3.71 (m, 2H), 3.36 (s, 1H), 2.60-2.58 (s, 3H), 2.35-2.07 (m, 2H), 1.19-1.12 (m, 9H).

[00599] Example of the synthesis of 1-(4-(azetidin-3-yl)piperazin-1-yl)-2-((4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)amino)ethane-1 -one hydrochloride

[00600] Step 1: Preparation of N-(4-chloro-2-methoxy-5-(1-methylcyclopropyl)phenyl)acetamide

[00601] N-(4-chloro-2-methoxy-5-(1-methylcyclopropyl)phenyl)acetamide was prepared according to the procedure set forth in US Patent Application Publication No. 2014/288045 A1, which is incorporated herein by reference in in full.

[00602] Step 2: Preparation of N-(4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)acetamide

[00603] To a solution of N-(4-chloro-2-methoxy-5-(1-methylcyclopropyl)phenyl)acetamide (2.0 g, 7.88 mmol, 1.0 eq.) in EtSH (20 ml) was added AlCl ₃ (10.5 g, 78.8 mmol, 1.0 eq.) at 20°C. After stirring for 20 hours, the mixture was quenched with ice water. The pH was adjusted to ~9 with aqueous NaHCO ₃ and the mixture was extracted with EtOAc (ethyl ethanoate). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo. The residue was purified by silica gel column chromatography with PE:EtOAc (1:1) to give the desired product N-(4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)acetamide (1.2 g, yield: 63 .6%) in the form of a yellow solid. LC/MS (ESI) m/z: 241.1 [M+1] ⁺ .

[00604] Step 3: Preparation of 2-amino-5-chloro-4-(1-methylcyclopropyl)phenol

[00605] A solution of N-(4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)acetamide (0.4 g, 2.51 mmol, 1.0 eq.) in EtOH/conc. HCl (27.5 ml, 10/1, v/v) was stirred at 100°C for 5 hours. After cooling to room temperature, the mixture was removed in vacuo to give the desired product 2-amino-5-chloro-4-(1-methylcyclopropyl)phenol (0.4 g, 100% yield) as a brown solid. LC/MS (ESI) m/z: 198.1 [M+1] ⁺ .

[00606] Step 4: Preparation of ethyl (4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)glycinate

[00607] AcOH (3 drops) was added to a solution of 2-amino-5-chloro-4-(1-methylcyclopropyl)phenol (0.4 g, 2.0 mmol, 1.0 eq.) in MeOH (20 mL) and ethylglyoxalate (306 mg, 3.0 mmol, 50% in toluene) at 20°C. After stirring for 2 hours, NaBH ₃ CN (378 mg, 6.0 mmol) was added. The resulting solution was stirred at 50°C for 20 hours. The reaction mixture was then poured into ethyl acetate and aqueous solution. The organic phase was dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica gel column chromatography with PE:EtOAc (10:1) to give the desired product ethyl (4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)glycinate (0.4 g, yield: 70. 5%) as a yellow oil. LC/MS (ESI) m/z: 284.1 [M+1] ⁺ .

[00608] Step 5: Preparation of (4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)glycine

[00609] To a solution of ethyl (4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)glycinate (0.4 g, 1.48 mmol, 1.0 eq.) in THF (tetrahydrofuran, tetrahydrofuran) / H ₂ O (16 ml, 3/1, v/v) was added with LiOH (620 mg, 14.8 mmol) at 25°C. After stirring for 1 hour, the pH was adjusted to ~2 with 1 M HCl. The mixture was treated with EtOAc. The organic phase was dried over Na ₂ SO ₄ , filtered and concentrated to give (4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)glycine (0.18 g) as a brown oil. ¹ H-NMR (400 MHz, CD ₃ OD) δ 6.66 (s, 1H), 6.25 (s, 1H), 5.38 (q, J=6.4 Hz, 1H), 3.86 (s, 2H), 1.90 (s, 3H), 1.72 (dd, J=6.8 Hz, 3H).

[00610] Step 6: Preparation of benzyl 4-(1-(tert-butoxycarbonyl)azetidin-3-yl)piperazine-1-carboxylate

[00611] AcOH was added to a solution of benzylpiperazine-1-carboxylate (10 g, 45.4 mmol) and tert-butyl-3-oxoazetidine-1-carboxylate (8.5 g, 49.6 mmol) in MeOH (200 mL). (3 ml). After stirring at 25°C for 1 hour, NaBH ₃ CN (8.5 g, 136.2 mmol) was added to the mixture and the resulting mixture was stirred at 25°C for 16 hours. The mixture was quenched with aqueous NaHCO ₃ and extracted with EtOAc. The organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography with PE:EtOAc (5:1) to give benzyl 4-(1-(tert-butoxycarbonyl)azetidin-3-yl)piperazine-1-carboxylate (12.6 g, yield: 75%) as a white solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.38-7.30 (m, 5H), 5.13 (s, 2H), 4.17-4.09 (m, 2H), 3.94 -3.90 (m, 2H), 3.59-3.52 (m, 4H), 3.09-3.04 (m, 1H), 2.30 (s, 4H), 1.43 (s , 9Н).

[00612] Step 7: Preparation of tert-butyl-3-(piperazin-1-yl)azetidine-1-carboxylate

[00613] Pd/C ( 0.5 g, 10%). The resulting mixture was stirred under H2 atmosphere at 25°C for 20 hours. The mixture was filtered and evaporated under reduced pressure to obtain tert-butyl-3-(piperazin-1-yl)azetidin-1-carboxylate (950 mg, yield: 87%) as a colorless oil.

[00614] Step 8: Preparation of tert-butyl 3-(4-((4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)glycyl)piperazin-1-yl)azetidine-1-carboxylate

[00615] DIEA (530 mg, 4.1 mmol), (4-chloro-2- hydroxy-5-(1-methylcyclopropyl)phenyl)glycine (290 mg, 1.13 mmol) and Benzotriazol-1-yl-oxytripyrrolidinophosphoniinn hexafluorophosphate, benzotriazol-1-yl-oxytripyrrolidinophosphonite hexafluorophosphate (1.07 g, 2. 06 mmol). The mixture was stirred at 20°C for 1 hour. The mixture was quenched with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography with DCM:MeOH (50:1 ~ 20:1) to give tert-butyl-3-(4-((4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl) glycyl)piperazin-1-yl)azetidine-1-carboxylate (210 mg, yield: 43%) as a brown oil. LC/MS (ESI) m/z: 479.2 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 6.84-6.79 (m, 1H), 6.43 (s, 1H), 3.96-3.88 (m, 4H), 3.82 -3.79 (m, 2H), 3.71 (s, 2H), 3.48 (d, J=4.0 Hz, 2H), 3.11-3.08 (m, 1H), 2, 36 (d, J=4.0 Hz, 4H), 1.93 (s, 2H), 1.82-1.73 (m, 3H), 1.41 (s, 9H), 1.26-1 .23 (m. 2H).

[00616] Step 9: Preparation of 1-(4-(azetidin-3-yl)piperazin-1-yl)-2-((4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)amino)ethane -1-one hydrochloride

[00617] Tert-Butyl-3-(4-((4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)glycyl)piperazin-1-yl)azetidine-1-carboxylate solution (210 mg, 0 .44 mmol) in dioxane (2 ml) and HCl (6 M in dioxane, 1 ml). The resulting mixture was stirred at a temperature of 20°C for 0.5 hours. The mixture was concentrated to give 1-(4-(azetidin-3-yl)piperazin-1-yl)-2-((4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)amino)ethane-1- it hydrochloride (120 mg, 66% yield) as a white solid. LC/MS (ESI) m/z: 379.2 [M+1] ⁺ .

[00618] Example of the synthesis of tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-((S)-2-(4-(3-hydroxybenzoyl)thiazol-2-yl)pyrrolidine) -1-yl)-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate

[00619] Step 1: Preparation of tert-butyl (S)-2-carbamoylpyrrolidine-1-carboxylate

[00620] A solution of methyl 2-chloroacetate (30.0 g, 245 .9 mmol) in tetrahydrofuran (40 ml) at minus 10°C. After stirring for 30 minutes, ammonium hydroxide (30 ml) was added to the reaction mixture at minus 10°C. The resulting mixture was stirred at room temperature overnight. The volatile components were evaporated under reduced pressure to obtain a residue, which was treated with ethyl acetate (100 ml), washed with aqueous sodium bicarbonate (saturated, 40 ml), then with brine (40 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give tert-butyl (S)-2-carbamoylpyrrolidine-1-carboxylate (22.0 g) as a colorless oil.

[00621] Step 2: Preparation of tert-butyl (S)-2-carbamothioylpyrrolidine-1-carboxylate

[00622] A mixture of (S)-2-carbamoylpyrrolidine-1-carboxylate (22.0 g) in tetrahydrofuran (80 ml) and Lawesson's reagent (41.8 g, 103.40 mmol) was stirred at 70°C for 2 hours . The mixture was partitioned between ethyl acetate (400 ml) and water (400 ml). The organic layer was collected, washed with brine (300 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by flash chromatography on silica gel (eluting with 20% ethyl acetate in hexane) to give tert-butyl(S )-2-carbamothioylpyrrolidine-1-carboxylate (8.3 g, 38% yield) as a white solid. LC/MS (ESI) m/z: 231.0 [M+1] ⁺ .

[00623] Step 3: Preparation of ethyl(S)-2-(1-(tert-butoxycarbonyl)pyrrolidin-2-yl)triazol-4-carboxylate

[00624] A mixture of tert-butyl (S)-2-carbamothioylpyrrolidine-1-carboxylate (8.3 g, 40.4 mmol) and ethyl 3-bromo-2-oxopropanoate (13.8 g, 56.6 mmol) in ethanol (80 ml) was stirred at 60°C for 2 hours. The volatile components were evaporated under reduced pressure to obtain a crude residue, which was dissolved in a saturated aqueous solution of sodium bicarbonate (10.6 g, 100 mmol in 80 ml water)-tetrahydrofuran (80 ml). Di-tert-butyl dicarbonate (8.81 g, 40.4 mmol) was added to the resulting mixture and stirred at room temperature for 2 hours. The volatile components were evaporated under reduced pressure and the resulting aqueous solution was extracted with ethyl acetate (300 ml). The organic layer was collected, washed with brine (150 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by flash silica gel chromatography (eluting with 20% ethyl acetate in hexane) to give ethyl(S)- 2-(1-(tert-butoxycarbonyl)pyrrolidin-2-yl)thiazol-4-carboxylate (6.8 g, 52% yield) as a colorless oil. LC/MS (ESI) m/z: 327.0 [M+1] ⁺ .

[00625] Step 4: Preparation of (S)-2-(1-(tert-butoxycarbonyl)pyrrolidin-2-yl)triazole-4-carboxylic acid

[00626] A mixture of ethyl (S)-2-(1-(tert-butoxycarbonyl)pyrrolidin-2-yl)thiazol-4-carboxylate (6.8 g, 20.90 mmol) and sodium hydroxide (1.67 g, 41.8 mmol) in methanol (30 ml)-water (30 ml) was stirred at room temperature for 1 hour. Volatile components were evaporated under reduced pressure. The resulting aqueous solution was acidified with aqueous hydrochloric acid (1 N) to pH 3-4 and extracted with ethyl acetate (60 ml). The organic layer was collected, washed with brine (20 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give (S)-2-(1-(tert-butoxycarbonyl)pyrrolidin-2-yl)thiazol-4-carboxylic acid (4.5 g, 72% yield) as a white solid. LC/MS (ESI) m/z: 299.0 [M+1] ⁺ .

[00627] Step 5: Preparation of tert-butyl (S)-2-(4-(methoxy(methyl)carbamoyl)thiazol-2-yl)pyrrolidin-1-carboxylate

[00628] To a mixture of (S)-2-(1-(tert-butoxycarbonyl)pyrrolidin-2-yl)thiazol-4-carboxylic acid (3.88 g, 15.10 mmol), N,N-dimethylhydroxylamine hydrochloride ( 1.53 g, 15.60 mmol) and N-ethyl-N-isopropylpropan-2-amine (5.03 g, 39.0 mmol) in N,N-dimethylformamide (20 ml) 1-ethyl-3 hydrochloride was added -(3-dimethylaminopropyl)carbodiimide (3.25 g, 16.90 mmol) and 1-hydroxybenzotriazole (2.11 g, 15.60 mmol) at 0°C, the resulting mixture was allowed to warm to room temperature and stirred at room temperature within 12 hours. The mixture was partitioned between ethyl acetate (200 ml) and water (100 ml). The organic layer was collected, washed with water (100 ml×2), then with brine (100 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue, which was purified by flash chromatography on silica gel (eluting with 20% ethyl acetate in hexane) to give tert-butyl(S)-2-(4-(methoxy(methyl)carbamoyl)thiazol-2-yl)pyrrolidine-1-carboxylate (3.46 g, 78% yield) as a colorless oil. LC/MS (ESI) m/z: 342.0 [M+1] ⁺ .

[00629] Step 6: Preparation of tert-butyl (S)-2-(4-(3-methoxybenzoyl)thiazol-2-yl)pyrrolidin-1-carboxylate

[00630] To a mixture of tert-butyl (S)-2-(4-(methoxy(methyl)carbamoyl)thiazol-2-yl)pyrrolidine-1-carboxylate (3.3 g, 9.46 mmol) in anhydrous tetrahydrofuran ( 30 ml) after adding (3-methoxyphenyl)magnesium bromide (28.4 ml, 28.40 mmol) dropwise at minus 40°C, the mixture was stirred at minus 40°C for 1 hour. The mixture was quenched with ammonium chloride solution (100 ml) at -0°C. The resulting mixture was extracted with ethyl acetate (30 ml x 3). The organic layers were collected, washed with brine (80 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by flash chromatography on silica gel (eluting with 20% ethyl acetate in hexane) to give (S)-tert -butyl 2-(4-(3-methoxybenzoyl)thiazol-2-yl)pyrrolidin-1-carboxylate (3.49 g, 95% yield) as a colorless oil. LC/MS (ESI) m/z: 389.1 [M+1] ⁺ .

[00631] Step 7: Preparation of (S)-(3-hydroxyphenyl)(2-(pyrrolidin-2-yl)thiazol-4-yl)methanone.

[00632] To a mixture of tert-butyl (S)-2-(4-(methoxy(methyl)carbamoyl)thiazol-2-yl)pyrrolidine-1-carboxylate (3.49 g, 8.99 mmol) in dichloromethane (20 ml) a solution of tribromoborane (4.2 ml) in dichloromethane (10 ml) was added at minus 45°C, the mixture was stirred at minus 45°C for 1 hour. The mixture was quenched with methanol (1 ml) at minus 78°C. After warming to room temperature, the resulting mixture was partitioned between dichloromethane (30 ml) and water (30 ml). The organic layer was collected, washed with brine (15 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by flash chromatography on silica gel (eluted with 5% methanol in dichloromethane) to give (S)-tert -butyl 2-(4-(3-methoxybenzoyl)thiazol-2-yl)pyrrolidin-1-carboxylate (1.25 g, 51% yield) as a colorless oil. LC/MS (ESI) m/z: 275.1 [M+1] ⁺ .

[00633] Step 8: Preparation of tert-butyl ((S)-1-cyclohexyl-2-((S)-2-(4-(3-hydroxybenzoyl)thiazol-2-yl)pyrrolidin)-1-yl)- 2-oxoethyl)carbamate

[00634] To a mixture of (S)-tert-butyl 2-(4-(3-methoxybenzoyl)thiazol-2-yl)pyrrolidine-1-carboxylate (1.20 g, 4.38 mmol), (S)-2 -((tert-butoxycarbonyl)amino)-2-cyclohexylacetic acid (2.81 g, 10.95 mmol) and N-ethyl-N]-isopropylpropan-2-amine (5.65 g, 43.80 mmol) in N,N-dimethylformamide (10 ml) was added 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.52 g, 13.14 mmol), 1-hydroxybenzotriazole (1.77 g, 13.14 mmol) at 0°C, the resulting mixture was allowed to warm to room temperature and stirred at room temperature for 1 hour. The mixture was partitioned between ethyl acetate (30 ml) and water (30 ml). The organic layer was collected, washed with brine (15 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue, which was dissolved in methanol, potassium carbonate (1.51 g, 10.95 mmol) was added, the mixture was stirred at room temperature for 2 hours. The mixture was filtered and concentrated under reduced pressure to give a crude residue, which was purified by flash chromatography on silica gel (eluting with 20% ethyl acetate in hexane) to give tert-butyl ((S)-1-cyclohexyl-2-((S)- 2-(4-(3-hydroxybenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)carbamate (1.35 g, 60% yield) as a colorless oil. LC/MS (ESI) m/z: 514.2 [M+1] ⁺ .

[00635] Step 9: Preparation of (S)-2-amino-2-cyclohexyl-1-((S)-2-(4-(3-hydroxybenzoyl)thiazol-2-yl)pyrrolidin-1-yl)ethane- 1-she

[00636] To a mixture of tert-butyl ((S)-1-cyclohexyl-2-((S)-2-(4-(3-hydroxybenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl )carbamate (1.34 g, 2.61 mmol) in dichloromethane (5 ml), 1 M hydrochloride in dioxane (1.0 ml) was added and the mixture was stirred at room temperature for 1.5 hours. The volatile components were evaporated under reduced pressure to leave a residue, which was made basic with saturated aqueous sodium bicarbonate and extracted with dichloromethane (20 ml x 2). The organic layers were collected, washed with brine (15 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give (S)-2-amino-2-cyclohexyl-1-((S)-2-(4-(3 -hydroxybenzoyl)thiazol-2-yl)pyrrolidin-1-yl)ethanone (980 mg) as a light yellow oil. LC/MS (ESI) m/z: 414.1 [M+1] ⁺ .

[00637] Step 10: Preparation of tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-((S)-2-(4-(3-hydroxybenzoyl)thiazol-2-yl) pyrrolidin)-1-yl)-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate

[00638] To a mixture of (S)-2-amino-2-cyclohexyl-1-((S)-2-(4-(3-hydroxybenzoyl)thiazol-2-yl)pyrrolidin-1-yl)ethanone (980 mg , 2.37 mmol) in dichloromethane (10 ml), N-ethyl-N-isopropylpropan-2-amine (5.03 g, 39.0 mmol) was added to a solution of tert-butyl (S)-(1-(2, 5-dioxopyrrolidin-1-yl)-1-oxopropan-2-yl)(methyl)carbamate (673 mg, 2.37 mmol) in dichloromethane (10 ml) at room temperature. The resulting mixture was stirred for 1 hour. The mixture was diluted with dichloromethane (30 ml) and washed with water (30 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by flash chromatography on silica gel (eluting with 20% ethyl acetate in hexane) to give tertiary butyl ((S)-1-(((S)-1-cyclohexyl-2-((S)-2-(4-(3-hydroxybenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2 -oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate (895 mg, 62% yield) as a white solid. LC/MS (ESI) m/z: 599.1 [M+1] ⁺ .

[00639] Example of the synthesis of tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-((S)-2-(4-(4-hydroxynaphthalene-1-yl)thiazol-2) -yl)pyrrolidin-1-yl)-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate

[00640] Step 1: Preparation of methyl (S)-2-((S)-2-((tert-butoxycarbonyl)(methyl)amino)propanamido)-2-cyclohexyl acetate

[00641] To a mixture of methyl (2S)-2-amino-2-cyclohexyl acetate (1.60 g, 7.70 mmol, 1.00 eq., hydrogen chloride) and (2S)-2-[tert-butoxycarbonyl(methyl )amino]propanoic acid (1.57 g, 7.70 mmol, 1.00 eq.) in dichloromethane (20 ml) was added O-(7-azabenzotriazol-1-yl)-N,N,N',N' -tetramethyluronium hexafluorophosphate (4.39 g, 11.56 mmol, 1.50 eq.) and triethylamine (2.34 g, 23.11 mmol, 3.22 mL, 3.00 eq.) per serving. The mixture was stirred at 30°C for 12 hours. The reaction mixture was washed with water (50 ml x 2) and the organic phase was concentrated in vacuo. The residue was purified by column chromatography (petroleum ether/ethyl acetate 5/1) to give methyl(2S)-2-[[(2S)-2-[tert-butoxycarbonyl(methyl)amino]propanoyl]amino]-2- cyclohexyl acetate (2.40 g, 6.73 mmol, 87% yield) as a white solid. LC/MS (ESI) m/z: 357.2 [M+1] ⁺ .

[00642] Step 2: Preparation of (S)-2-((S)-2-((tert-butoxycarbonyl)(methyl)amino)propanamido)-2-cyclohexylacetic acid

[00643] To a mixture of methyl (2S)-2-[[(2S)-2-[tert-butoxycarbonyl(methyl)amino]propanoyl]amino]-2-cyclohexyl acetate (2.40 g, 6.73 mmol, 1. 00 eq.) in tetrahydrofuran (15 ml), a mixture of lithium hydroxide (0.42 g, 10.10 mmol, 1.50 eq.) in water (5 ml) was added in portions at 0° C. under a nitrogen atmosphere. The mixture was stirred at 20°C for 1 hour. The mixture was diluted with water (50 ml), extracted with ethyl acetate (50 ml × 2). The aqueous phase was adjusted to pH approximately 5 with hydrogen chloride solution (1 M). The mixture was then extracted with ethyl acetate (40 ml x 3). The combined organic layer was washed with water (80 ml x 2) and brine (80 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the product. The compound (2S)-2-[[(2S)-2-[tert-butoxycarbonyl(methyl)amino]propanoyl]amino]-2-cyclohexylacetic acid (2.20 g, 6.42 mmol, 95% yield) was obtained in as a white solid. LC/MS (ESI) m/z: 287.1 [M-55] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.01-6.55 (m, 1H), 4.74 (br s, 1H), 4.52 (br dd, J=5.0, 8, 3 Hz, 1H), 2.81 (d, J=3.3 Hz, 3H), 1.95-1.81 (m, 1H), 1.79-1.59 (m, 4H), 1, 49 (s, 8H), 1.40-1.18 (m, 5H), 1.16-0.96 (m, 3H).

[00644] Step 3: Preparation of 1-(4-fluoronaphthalene-1-yl)ethan-1-one

[00645] To a mixture of 1-fluoronaphthalene (5.00 g, 34.21 mmol, 4.42 mL, 1.00 eq.) and aluminum chloride (6.84 g, 51.31 mmol, 2.80 mL, 1 .50 eq.) in dichloromethane (50 ml) was added acetyl chloride (3.22 g, 41.05 mmol, 2.93 ml, 1.20 eq.) in dichloromethane (10 ml) at 0°C. The mixture was stirred at 0°C for 0.5 hours. The mixture was then stirred at 20°C for 4 hours. The mixture was poured into water (60 ml), extracted with dichloromethane (50 ml × 2). The combined organic layer was washed with water (80 ml×2) and brine (80 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (petroleum ether/ethyl acetate 1/0 to 20/1) to give the compound 1-(4-fluoro-1-naphthyl)ethanone (5.40 g, 28.69 mmol, 84% yield ) in the form of a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.94-8.86 (m, 1H), 8.17 (d, J=8.3 Hz, 1H), 7.98 (dd, J=5 ,4, 8.1 Hz, 1H), 7.71-7.65 (m, 1H), 7.64-7.58 (m, 1H), 7.16 (dd, J=8,l, 9 .7 Hz, 1H), 2.84-2.67 (m, 3H).

[00646] Step 4: Preparation of 1-(4-(benzyloxy)naphthalene-1-yl)ethan-1-one

[00647] To a mixture of phenylmethanol (4.48 g, 41.45 mmol, 4.31 mL, 1.50 eq.) in dimethylformamide (60 mL) was added potassium t-butoxide (4.65 g, 41.45 mmol, 1.50 eq.). The mixture was stirred at 20°C for 0.5 hour. 1-(4-fluoro-1-naphthyl)ethanone (5.20 g, 27.63 mmol, 1.00 eq.) was then added to the mixture. The mixture was stirred at 20°C for 2 hours. The mixture was diluted with water (200 ml), extracted with ethyl acetate (100 ml × 2). The combined organic layer was washed with water (100 ml x 2) and brine (100 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (petroleum ether/ethyl acetate 30/1 to 10/1) to give the compound 1-(4-benzyloxy-1-naphthyl)ethanone (7.64 g, 22.80 mmol, 83% yield ) as a white solid.

[00648] Step 5: Preparation of 1-(4-(benzyloxy)naphthalene-1-yl)-2-bromoethan-1-one

[00649] Liquid bromine (2.89 g, 18 .09 mmol, 932.78 µl, 1.00 equiv) in portions at 20°C in an atmosphere of N ₂ . The mixture was stirred for 2 hours. The reaction mixture was quenched by adding saturated sodium thiosulfate (50 ml) and the organic phase was concentrated in vacuo. The residue was purified by column chromatography (petroleum ether/ethyl acetate 30/1) to give 1-(4-benzyloxy-1-naphthyl)-2-bromoethanone (4.50 g, 12.67 mmol, 70% yield) in as a white solid. LC/MS (ESI) m/z: 354.9 [M+1] ⁺ .

[00650] Step 6: Preparation of tert-butyl (S)-2-(4-(4-(benzyloxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-carboxylate

[00651] To a mixture of 1-(4-benzyloxy-1-naphthyl)-2-bromoethanone (3.90 g, 10.98 mmol, 1.00 eq.) and tert-butyl (2S)-2-carbamothioylpyrrolidine-1 -carboxylate (3.79 g, 16.47 mmol, 1.50 eq.) in ethyl alcohol (80 ml) was added pyridine (0.87 g, 10.98 mmol, 886.16 μl, 1.00 eq.) in a nitrogen atmosphere. The mixture was heated to a temperature of 80°C and stirred for 1 hour. The reaction mixture was concentrated in vacuo. The residue was purified by column chromatography (petroleum ether/ethyl acetate 5/1) to give tert-butyl (2S)-2-[4-(4-benzyloxy-1-naphthyl)thiazol-2-yl]pyrrolidin-1- carboxylate (5.30 g, 10.89 mmol, 99% yield) as a white solid. LC/MS (ESI) m/z: 487.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.49-8.37 (m, 1H), 8.35-8.31 (m, 1H), 8.23 (br s, 1H), 7, 61-7.48 (m, 5H), 7.47-7.33 (m, 3H), 7.27 (s, 1H), 7.26 (br s, 1H), 6.93 (d, J =7.9 Hz, 1H), 5.32 (s, 2H), 3.77-3.42 (m, 2H), 2.48-2.25 (m, 2H), 2.14-1, 94 (m, 3H), 1.57-1.34 (m, 9H).

[00652] Step 7: Preparation of (S)-4-(4-(benzyloxy)naphthalene-1-yl)-2-(pyrrolidin-2-yl)thiazole

[00653] To a mixture of tert-butyl (2S)-2-[4-(4-benzyloxy-1-naphthyl)thiazol-2-yl]pyrrolidin-1-carboxylate (2.00 g, 4.11 mmol, 1. 00 eq.) in dichloromethane (36 mL), trifluoroacetic acid (18.12 g, 158.89 mmol, 11.76 mL, 38.66 eq.) was added in one portion under N ₂ atmosphere. The mixture was stirred at 30°C for 1 hour. The reaction mixture was concentrated in vacuo. The compound 4-(4-benzyloxy-1-naphthyl)-2-[(2S)-pyrrolidin-2-yl]thiazole (2.06 g, trifluoroacetic acid) was obtained as a white solid. LC/MS (ESI) m/z: 387.2 [M+1] ⁺ .

[00654] Step 8: Preparation of tert-butyl ((S)-1-(((S)-2-((S)-2-(4-(4-(benzyloxy)naphthalene-1-yl)thiazole)2 -yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate

[00655] To a mixture of (2S)-2-[[(2S)-2-[tert-butoxycarbonyl(methyl)amino]propanoyl]amino]-2-cyclohexylacetic acid (1.15 g, 3.36 mmol, 1 eq) 4-(4-benzyloxy-1-naphthyl)-2-[(2S)-pyrrolidin-2-yl]thiazole (2.02 g, 4.03 mmol, 1.2 eq, trifluoroacetic acid) and 4-methylmorpholine (1.36 g, 13.43 mmol, 1.48 ml, 4 eq.) in tetrahydrofuran (40 ml) and dimethylformamide (4 ml) was added 5-triazin-2-yl)-4-methylmorpholine-4-ium; chloride; hydrate (1.98 g, 6.72 mmol, 2 eq.). The mixture was stirred at 20°C for 12 hours. The reaction mixture was quenched by adding water (30 ml) and extracted with ethyl acetate (30 ml x 3), then the organic phase was concentrated in vacuo. The residue was purified by column chromatography (petroleum ether/ethyl acetate 5/1) to give tert-butylN-[(1S)-2-[[(1S)-2-[(2S)-2-[4-(4 -benzyloxy-1-naphthyl)thiazol-2-yl]pyrrolidin-1-yl]-1-cyclohexyl-2-oxoethyl]amino]-1-methyl-2-oxoethyl]-N-methyl-carbamate (0.80 g , 1.09 mmol, yield 33%, purity 97%) as a white solid. LC/MS (ESI) m/z: 711.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.46-8.39 (m, 1H), 8.25-8.18 (m, 1H), 8.03 (s, 1H), 7.59 -7.49 (m, 5H), 7.47 -7.34 (m, 3H), 7.25 (s, 1H), 6.92 (d, J=8.l Hz, 1H), 5, 70-5.61 (m, 1H), 4.79-4.59 (m, 2H), 3.96-3.78 (m, 2H), 2.97 (s, 2H), 2.89 ( s, 2H), 2.82 (s, 3H), 2.47 (br s, 1H), 2.37-2.16 (m, 2H), 2.05 (s, 2H), 1.88- 1.54 (m, 17H), 1.50 (s, 9H), 1.53-1.45 (m, 1H), 1.35 (d, J=7.l Hz, 3H), 1.30 -0.93 (m, 9H).

[00656] Step 9: Preparation of tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-((S)-2-(4-(4-hydroxynaphthalene-1-yl)thiazol- 2)-yl)pyrrolidin-1-yl)-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate

[00657] To a mixture of tert-butyl N-[(1S)-2-[[(1S)-2-[(2S)-2-[4-(4-benzyloxy-1-naphthyl)thiazol-2-yl] pyrrolidin-1-yl]-1-cyclohexyl-2-oxoethyl]amino]-1-methyl-2-oxoethyl]-N-methylcarbamate (0.50 g, 0.70 mmol, 1.00 eq.) in methyl alcohol (10 mL), palladium hydroxide (0.36 g, 2.53 mmol, 3.60 eq.) and hydrogen (0.01 g, 0.70 mmol, 1.00 eq.) were added in one portion under nitrogen. The mixture was stirred at 20°C for 16 hours. The reaction mixture was filtered and the filtrate was concentrated in vacuo. The compound tert-butyl N-[(1S)-2-[[(1S)-1-cyclohexyl-2-[(2S)-2-[4-(4-hydroxy-1-naphthyl)thiazol-2-yl was obtained ]pyrrolidin-1-yl]-2-oxoethyl]amino]-1-methyl-2-oxoethyl]-N-methylcarbamate (0.3 g, 0.48 mmol, 69% yield) as a white solid. LC/MS (ESI) m/z: 621.3 [M+1] ⁺ .

[00658] Example of the synthesis of tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-((S)-2-(4-(4-(2-(2-(2-( methylamino)ethoxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate

[00659] Step 1: Preparation of tert-butyl ((S)-1-(((S)-2-((S)-2-(4-(4-(2-(2-(2-bromoethoxy)ethoxy )ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate

[00660] To a mixture of tert-butyl N-[(1S)-2-[[(1S)-1-cyclohexyl-2-[(2S)-2-[4-(4-hydroxy-1-naphthyl)thiazol- 2-yl]pyrrolidin-1-yl]-2-oxoethyl]amino]-1-methyl-2-oxoethyl]-N-methylcarbamate (0.30 g, 0.48 mmol, 1.00 eq.) and 1, 2-bis(2-bromoethoxy)ethane (0.27 g, 0.97 mmol, 2.00 eq.) in acetonitrile (12 ml) was added potassium carbonate (0.23 g, 1.69 mmol, 3.50 eq. .) in a nitrogen atmosphere. The mixture was heated to a temperature of 80°C and stirred for 12 hours. The reaction mixture was washed with water (10 ml) and extracted with ethyl acetate (20 ml), then the organic phase was concentrated in vacuo. The residue was purified by column chromatography (petroleum ether/ethyl acetate 1/1) to give tert-butyl N-[(1S)-2-[[(1S)-2-[(2S)-2-[4-[ 4-[2-[2-(2-bromoethoxy)ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-1-cyclohexyl-2-oxoethyl]amino]-1-methyl- 2-oxoethyl]-N-methylcarbamate (0.15 g, 0.18 mmol, 38% yield) as a white solid. LC/MS (ESI) m/z: 817.3 [M+1] ⁺ .

[00661] Step 2: Preparation of tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-((S)-2-(4-(4-(2-(2-(2 -(methylamino)ethoxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)amino)-1-oxopropan-2-l)(methyl)carbamate

[00662] Mixture of tert-butyl N-[(1S)-2-[[(1S)-2-[(2S)-2-[4-[4-[2-[2-(2-bromoethoxy)ethoxy] ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-1-cyclohexyl-2-oxoethyl]amino]-1-methyl-2-oxoethyl]-N-methyl-carbamate (0.14 g , 0.17 mmol, 1.00 eq.) and methylamine/ethyl alcohol (10 ml) were heated to 60°C and stirred for 12 hours. The reaction mixture was concentrated in vacuo. The compound tert-butyl N-[(1S)-2-[[(1S)-1-cyclohexyl-2-[(2S)-2-[4-[4-[2-[2-[2-(methylamino) was obtained )ethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-2-oxoethyl]amino]-1-methyl-2-oxoethyl]-N-methylcarbamate (0.13 g) as a white solid. LC/MS (ESI) m/z: 766.4 [M+1] ⁺ .

[00663] Example of the synthesis of tert-butyl (2-(2-(2-aminoethoxy)ethoxy)ethyl)(methyl)carbamate

[00664] Step 1: Preparation of tert-butyl (2-(2-(2-(1,3-dioxyisoindolin-2-yl)ethoxy)ethoxy)ethyl)(methyl)carbamate

[00665] To a solution of 2-[2-[2-[tert-butoxycarbonyl(methyl)amino]ethoxy]ethoxy]ethyl 4-methylbenzenesulfonate (400 mg, 0.96 mmol, 1 eq.) in N,N-dimethylformamide ( 10 ml) (1,3-dioxoisoindolin-2-yl)potassium (213 mg, 1.15 mmol, 1.2 eq.) was added. The mixture was stirred at 50°C for 12 hours. The mixture was diluted with water (20 ml) and the aqueous phase was extracted with ethyl acetate (20 ml x 3). The combined organic phase was washed with brine (20 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative TLC (petroleum ether/ethyl acetate 3/1) to give tert-butyl-N-[2-[2-[2-(1,3-dioxoisoindolin-2-yl)ethoxy]ethoxy]ethyl] -N-methylcarbamate (300 mg, 0.76 mmol, 79% yield) as a colorless oil. LC/MS (ESI) m/z: 415.1 [M+23] ⁺ .

[00666] Step 2: Preparation of tert-butyl (2-(2-(2-aminoethoxy)ethoxy)ethyl)(methyl)carbamate

[00667] To a solution of tert-butyl-N-[2-[2-[2-(1,3-dioxoisoindolin-2-yl)ethoxy]ethoxy]ethyl]-N-methylcarbamate (300 mg, 0.76 mmol, 1 equiv.) in ethanol (5 ml) was added hydrazine hydrate (382 mg, 7.64 mmol, 0.4 ml, 10 equiv.). The mixture was stirred at 80°C for 12 hours. The mixture was diluted with water (20 ml) and the aqueous phase was extracted with dichloromethane (20 ml x 3). The combined organic phase was washed with brine (20 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative TLC (dichloromethane/methanol 5/2) to give tert-butyl-1-N-[2-[2-(2-aminoethoxy)ethoxy]ethyl]-N-methylcarbamate (120 mg, 0. 46%) mmol, yield 59%) as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 3.66-3.56 (m, 6H), 3.55-3.49 (m, 2H), 3.40 (br s, 2H), 2, 99-2.79 (m, 5H), 1.74 (br s, 3H), 1.46 (s, 9H).

[00668] Synthesis example of (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-N-(2-methoxy- 4-((2-(2-(2-(methylamino)ethoxy)ethoxy)ethyl)carbamoyl)phenyl)-5-neopentylpyrrolidine-2-carboxamide

[00669] Step 1: Preparation of (Z)-3-(3-chloro-2-fluorophenyl)-2-(4-chloro-2-fluorophenyl)acrylonitrile.

[00670] To a mixture of 3-chloro-2-fluorobenzaldehyde (13.7 g, 86.68 mmol, 1.05 eq.) m 2-(4-chloro-2-fluorophenyl)acetonitrile (14 g, 82.56 mmol , 1 equiv.) in methanol (200 ml), a solution of sodium methoxide (13.4 g, 247.67 mmol, 3 equiv.) in methanol (40 ml) was added dropwise at 0°C under a nitrogen atmosphere. During addition, the product begins to precipitate. The suspension was stirred at 45°C for 5 hours. The solid was filtered and washed with water (200 ml) and methanol (50 ml) and then dried in vacuo to give (Z)-3-(3-chloro-2-fluorophenyl)-2-(4-chloro)-2 -fluorophenyl)prop-2-enenitrile (24 g, 77.39 mmol, 93% yield) as a white solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.20-8.10 (m, 1H), 7.81 (s, 1H), 7.64-7.49 (m, 2H), 7.33 -7.20 (m, 3H).

[00671] Step 2: Preparation of ethyl 4-(2-(((benzyloxy)carbonyl)amino)acetamido)-3-methoxybenzoate

[00672] To a mixture of 2-(benzyloxycarbonylamino)acetic acid (5.9 g, 28.17 mmol, 1.1 eq.) and ethyl 4-amino-3-methoxybenzoate (5 g, 25.61 mmol, 1 eq. ) in tetrahydrofuran (50 ml) was added 0-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (11.7 g, 30.74 mmol, 1.2 eq.) and N, N-Diisopropylethylamine (6.6 g, 51.23 mmol, 8.9 mL, 2 eq.) at 0°C under nitrogen. The mixture was heated to a temperature of 20°C and stirred for 12 hours. The mixture was diluted with water (100 ml) and extracted with ethyl acetate (100 ml x 3). The combined organic phase was washed with brine (200 ml x 2), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel (petroleum ether/ethyl acetate 10/1, 3/1 to 1/1) to give ethyl 4-[[2-(benzyloxycarbonylamino)acetyl]amino]-3-methoxybenzoate (9 g, 23.29 mmol, yield 91%) as a yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.41 (br d, J=8.4 Hz, 2H), 7.68 (br d, J=8.4 Hz, 1H), 7.53 ( s, 1H), 7.42-7.28 (m, 5H), 5.61 (br s, 1H), 5.18 (s, 2H), 4.42-4.32 (m, 2H), 4.11-4.01 (m, 2H), 3.88 (s, 3H), 1.40 (t, J=7.2 Hz, 3H).

[00673] Step 3: Preparation of ethyl 4-(2-aminoacetamido)-3-methoxybenzoate

[00674] To a solution of ethyl 4-[[2-(benzyloxycarbonylamino)acetyl]amino]-3-methoxybenzoate (8.8 g, 22.77 mmol, 1 eq.) in ethanol (100 ml) was added a palladium-activated carbon catalyst (1 g, purity 10%) under nitrogen atmosphere. The suspension was degassed in vacuum and purged with hydrogen several times. The mixture was stirred under a hydrogen atmosphere (15 psi) at 50°C for 24 hours. The mixture was then stirred under a hydrogen atmosphere (50 psi) at 50° C. for 5 hours. The reaction mixture was filtered and the filtrate was concentrated. The crude product was purified by silica gel chromatography (petroleum ether/ethyl acetate 10/1, 3/1 to 1/1) to give ethyl 4-[(2-aminoacetyl) amino]-3-methoxybenzoate (3.2 g, 12.69 mmol, 56% yield) as a yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 10.00 (br s, 1H), 8.53 (d, J=8.4 Hz, 1H), 7.71 (dd, J=1.6, 8.4 Hz, 1H), 7.57 (d, J=1.6 Hz, 1H), 4.38 (q, J=7.2 Hz, 2H), 3.97 (s, 3H), 3 .53 (s, 2H), 1.40 (t, J=7.2 Hz, 3H).

[00675] Step 4: Preparation of ethyl (E)-4-(2-((3,3-dimethylbutylidene)amino)acetamido)-3-methoxybenzoate

[00676] To a mixture of ethyl 4-[(2-aminoacetyl)amino]-3-methoxybenzoate (3.2 g, 12.69 mmol, 1 eq.) and 3,3-dimethylbutanal (1.3 g, 12.94 mmol, 1.6 mL, 1.02 eq.) in dichloromethane (50 mL) was added magnesium sulfate (3.0 g, 25.37 mmol, 2 eq.) at 20°C. The mixture was stirred for 20 hours. The mixture was filtered and the filtrate was concentrated in vacuo to give ethyl 4-[[2-[(E)-3,3-dimethylbutylideneamino]acetyl]amino]-3-methoxybenzoate (4.24 g, 12.68 mmol, 100% yield ) as a yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 9.45 (br s, 1H), 8.53 (d, J=8.4 Hz, 1H), 7.89-7.82 (m, 1H) , 7.72-7.68 (m, 1H), 7.58 -7.53 (m, 1H), 4.39-4.34 (m, 2H), 4.25-4.20 (m, 2H), 3.94 (s, 3H), 2.30-2.27 (m, 2H), 1.42-1.38 (m, 3H), 1.04 (s, 9H).

[00677] Step 5: Preparation of ethyl 4-((2R,3S,4R,5R)-3-(3-xnop-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano- 5-neopentylpyrrolidine-2-carboxamido)-3-methoxybenzoate

[00678] Copper acetoxy was added to a solution of [1-(2-diphenylphosphanyl-1-naphthyl)-2-naphthyl]diphenylphosphane (530 mg, 0.85 mmol, 0.12 eq.) in 2-methyltetrahydrofuran (10 ml) (95 mg, 0.78 mmol, 0.11 eq.). The mixture was stirred at 20°C for 30 minutes. Ethyl 4-[[2-[(E)-3,3-dimethylbutylideneamino]acetyl]amino]-3-methoxybenzoate (2.6 g, 7.80 mmol, 1.1 eq.) and (Z)-3- (3-chloro-2-fluorophenyl)-2-(4-chloro-2-fluorophenyl)prop-2-enenitrile (2.2 g, 7.09 mmol, 1 eq.) in 2-methyltetrahydrofuran (100 ml) was added at 20°C in a nitrogen atmosphere. Triethylamine (717 mg, 7.09 mmol, 1.0 mL, 1 eq.) was then added. The mixture was stirred at 20°C for 36 hours. The mixture was diluted with water (100 ml) and extracted with ethyl acetate (100 ml x 3). The combined organic phase was washed with brine (100 ml x 2), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (petroleum ether/ethyl acetate 10/1 to 5/1) to give ethyl 4-[[(2R,3S,4R,5R)-3-(3-chloro-2-fluoro- phenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3-methoxybenzoate (3 g, 4.65 mmol, yield 65%) as a yellow solid.

[00679] Step 6: Preparation of 4-((2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5 - neopentylpyrrolidine-2-carboxamido)-3-methoxybenzoic acid

[00680] To a mixture of ethyl 4-[[(2R,3S,4R,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5- (2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3-methoxybenzoate (3.0 g, 4.65 mmol, 1 eq.) in water (5 ml), tetrahydrofuran (5 ml) and methanol (5 ml) sodium hydroxide (937 mg, 23.44 mmol, 5.04 eq.) was added at 20°C. The mixture was stirred at 20°C for 12 hours. The mixture was then heated to a temperature of 50°C and stirred for another 12 hours. The mixture was diluted with water (20 ml) and acidified with 1N hydrochloric acid to pH=5. During addition, the product begins to precipitate. The solid was filtered and dried in vacuo to obtain the crude product. The solid was triturated with acetonitrile (100 ml) to give 4-[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3-methoxybenzoic acid (1.2 g, 1.91 mmol, 41% yield, 98% ee, 95% de, purity 98%) as a white solid. ¹ H-NMR (400 MHz, CD ₃ OD) δ 8.38 (d, J=8.4 Hz, 1H), 7.76-7.70 (m, 1H), 7.69-7.62 ( m, 2H), 7.44-7.30 (m, 3H), 7.28-7.19 (m, 2H), 4.79-4.76 (m, 1H), 4.64 (d, J=8.0 Hz, 1H), 4.16-4.04 (m, 2H), 3.98 (s, 3H), 1.76-1.66 (m, 1H), 1.36 (br d, J=14.0 Hz, 1H), 1.04 (s, 9H).

[00681] Step 7: Preparation of tert-butyl (2-(2-(2-(4-((2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4- chloro-2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamido)-3-methoxybenzamido)ethoxy)ethoxy)ethyl)(methyl)carbamate

[00682] To a mixture of 4-[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-( 2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3-methoxybenzoic acid (200 mg, 0.32 mmol, 1 eq.) and tert-butyl N-[2-[2-(2-aminoethoxy)ethoxy ]ethyl]-N-methylcarbamate (85 mg, 0.32 mmol, 1 eq.) in N,N-dimethylformamide (8 ml) was added 1-hydroxybenzotriazole (65 mg, 0.48 mmol, 1.5 eq.), N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (93 mg, 0.48 mmol, 1.5 eq.) and N,N-diisopropylethylamine (125 mg, 0.97 mmol, 0.17 ml, 3 eq. ). The mixture was stirred at 20°C for 12 hours. The mixture was quenched with water (25 ml). The mixture was then extracted with ethyl acetate (25 ml x 3). The combined organic layer was washed with water (35 ml×2) and brine (35 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TGS (dichloromethane:methanol = 10:1) to give tert-butyl N-[2-[2-[2-[[4-[[(2R,3S,4R,5S)-3-(3 -chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3-methoxybenzoyl]amino]ethoxy] ethoxy]ethyl]-N-methylcarbamate (230 mg, 0.26 mmol, 82% yield) as a light yellow solid. LC/MS (ESI) m/z: 860.4 [M+1] ⁺ .

[00683] Step 8: Preparation of (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-N-(2- methoxy-4-((2-(2-(2-(methylamino)ethoxy)ethoxy)ethyl)carbamoyl)phenyl)-5-neopentylpyrrolidine-2-carboxamide

[00684] To a mixture of tert-butyl N-[2-[2-[2-[[4-[[(2R,3S,4R,5S)-3-(3-xnop-2-fluorophenyl)-4-( 4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3-methoxybenzoyl]amino]ethoxy]ethoxy]ethyl]-N-methylcarbamate (230 mg , 0.26 mmol, 1 eq.) in dichloromethane (8 ml) was added trifluoroacetic acid (2.00 ml). The mixture was stirred at 20°C for 0.5 hour. The mixture was concentrated under reduced pressure to obtain the product. (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)-N was obtained -[2-methoxy-4-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethylcarbamoyl]phenyl]pyrrolidine-2-carboxamide (230 mg, trilfluoroacetate) as a light yellow oil. LC/MS (ESI) m/z: 760.3 [M+1] ⁺ .

[00685] Example of the synthesis of (2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)-1-(3-methyl-2-(3-methylisoxazol- 5-yl)butanoyl)pyrrolidine-2-carboxamide

[00686] Step 1: Preparation of 2-(3-methylisoxazol-5-yl)acetic acid

[00687] To a solution of 3,5-dimethylisoxazole (15 g, 154.46 mmol, 15 mL, 1 eq.) in tetrahydrofuran (150 mL), n-butyllithium (2.5 M, 77 mL, 1.25 eq.) at minus 78°C under nitrogen atmosphere, the mixture was stirred at minus 55°C for 30 minutes, and then the mixture was bubbled with carbon dioxide for 30 minutes, the mixture was stirred at 25°C for 1 hour. The mixture was quenched with saturated ammonium chloride solution (50 ml), and the mixture was extracted with ethyl acetate (50 ml). The aqueous phase was adjusted to pH=2 with aqueous hydrochloric acid (2 M), the mixture was extracted with ethyl acetate (50 ml, three times), the organic phase was dried with anhydrous sodium sulfate, filtered and the filtrate was concentrated to give 2-(3-methylisoxazole-5 -yl)acetic acid (10 g, 70.86 mmol, 46% yield) as a brown solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 12.74 (br s, 1H), 6.24 (s, 1H), 3.83 (s, 2H), 2.20 (s, 3H) .

[00688] Step 2: Preparation of methyl 2-(3-methylisoxazol-5-yl) acetate

[00689] Thionyl chloride (12.65 g, 106.29 mmol, 7.71 ml, 1.5 eq.) at 0°C, and the mixture was stirred at 50°C for 4 hours. The mixture was concentrated to give the crude product. The said crude product was diluted with ethyl acetate (200 ml) and washed with water (200 ml) and then with saturated aqueous sodium bicarbonate (50 ml) and then with brine (50 ml), the organic phase was dried with anhydrous (compound), filtered, and the filtrate condensed to give methyl 2-(3-methylisoxazol-5-yl) acetate (10 g, 64.45 mmol, 91% yield) as a brown oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 6.11 (s, 1H), 3.80 (s, 2H), 3.76 (s, 3H), 2.30 (s, 3H).

[00690] Step 3: Preparation of methyl 3-methyl-2-(3-methylisoxazol-5-yl)butanoate

[00691] To a solution of methyl 2-(3-methylisoxazol-5-yl)acetate (10 g, 64.45 mmol, 1 eq.) in tetrahydrofuran (100 ml) was added sodium hydride (3.87 g, 96.68 mmol , 60% purity, 1.5 eq.) at 0°C, and then 2-iodopropane (13.15 g, 77.34 mmol, 7.74 mL, 1.2 eq.) was added at 0°C, mixture stirred at 25°C for 2 hours. Additional 2-iodopropane (2.55 g, 15.00 mmol, 1.5 ml) was added and the mixture was stirred at 25°C for 10 hours. The mixture was quenched with aqueous hydrochloric acid (1 M, 300 ml) and the mixture was extracted with ethyl acetate (200 ml, three times), the organic phase was dried with anhydrous sodium sulfate, filtered and the filtrate was concentrated to give methyl-3-methyl-2-(3-methylisoxazole -5-yl)butanoate (13 g) as a brown oil.

[00692] Step 4: Preparation of 3-methyl-2-(3-methylisoxazol-5-yl) butanoic acid

[00693] To a solution of methyl 3-methyl-2-(3-methylisoxazol-5-yl)butanoate (12.7 g, 64.39 mmol, 1 eq.) in methanol (90 ml) and water (60 ml) sodium hydroxide was added. (12.88 g, 321.96 mmol, 5 eq.), the mixture was stirred at 25°C for 2 hours. The mixture was concentrated to remove methanol and then the residue was diluted with water (200 ml) and extracted with ethyl acetate (200 ml), the aqueous phase was adjusted to pH=3 with aqueous hydrochloric acid (2 M) and then the mixture was extracted with dichloromethane (200 ml, three times ). The organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the crude product as a brown oil. This crude product was purified by preparative flash HPLC, the acetonitrile fraction was removed and the residue was extracted with dichloromethane (300 ml x 5). The organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the product 3-methyl-2-(3-methylisoxazol-5-yl)butanoic acid (7.5 g, 40.94 mmol, 63% yield) as a white solid substances. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 6.26 (s, 1H), 3.58 (d, J=8.7 Hz, 1H), 2.33-2.23 (m, 1H ), 2.21 (s, 3H), 0.95 (d, J=6.7 Hz, 3H), 0.82 (d, J=6.8 Hz, 3H).

[00694] Step 5: Preparation of 2-hydroxy-4-(4-methylthiazol-5-yl)benzonitrile

[00695] To a solution of 4-bromo-2-hydroxybenzonitrile (15 g, 75.75 mmol, 1 eq.) and 4-methylthiazole (20.28 g, 204.53 mmol, 19 mL, 2.7 eq.) in N-methylpyrrolidone (150 ml), potassium acetate (22.30 g, 227.25 mmol, 3 eq.) and palladium acetate (1.70 g, 7.58 mmol, 0.1 eq.) were added and the mixture was stirred at 110 °C in a nitrogen atmosphere for 6 hours. The mixture was quenched with water (500 ml), the aqueous phase was extracted with ethyl acetate (300 ml x 3). The combined organic phase was washed with brine (200 ml, twice), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo, then methyl tert-butyl ether (500 ml) was added to the mixture, and the organic phase was washed with water (100 ml) and saline. solution (100 ml, twice). The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 3/1 to 1/1). The compound 2-hydroxy-4-(4-methylthiazol-5-yl)benzonitrile (11 g, 50.87 mmol, 67% yield) was obtained as a yellow solid.

[00696] Step 6: Preparation of 2-(aminomethyl)-5-(4-methylthiazol-5-yl)phenol

[00697] Lithium aluminum hydride (7.72 g, 203 .46 mmol, 4 eq.) at 0°C, the mixture was stirred at 50°C for 3 hours. The mixture was quenched with water (8 ml) at 0°C, and then 15% aqueous sodium hydroxide (8 ml) was added, followed by water (8 ml), anhydrous sodium sulfate (30 g). The mixture was stirred at 25°C for 30 minutes, filtered and dichloromethane/methanol (4/1, 50 ml) was added to the solid. The mixture was stirred at 25°C for 1 hour, filtered and the combined filtrate was concentrated to give 2-(aminomethyl)-5-(4-methylthiazol-5-yl)phenol (7 g, 31.78 mmol, 62% yield) in as a brown solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.82 (s, 1H), 6.90 (d, J=7.5 Hz, 1H), 6.52 (d, J=1.6 Hz, 1H), 6.25 (dd, J=1.7, 7.5 Hz, 1H), 3.59 (s, 2H), 2.41 (s, 3H).

[00698] Step 7: Preparation of tert-butyl (2S,4R)-4-hydroxy-2-((2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate

[00699] To a solution of 2-(aminomethyl)-5-(4-methylthiazol-5-yl)phenol (7 g, 31.78 mmol, 1 eq.) and (28,411)-1-tert-butoxycarbonyl-4-hydroxypyrrolidine -2-carboxylic acid (7.35 g, 31.78 mmol, 1 eq.) in dimethylformamide (70 ml), diisopropylethylamine (12.32 g, 95.33 mmol, 16.60 ml, 3 eq.) was added. and then HATU (13.29 g, 34.95 mmol, 1.1 eq.), the mixture was stirred at 25°C for 2 hours. Additionally, (2S,4R)-1-tert-butoxycarbonyl-4-hydroxypyrrolidine-2-carboxylic acid (7.35 g, 31.78 mmol, 1 eq.) and HATU (12.08 g, 31.78 mmol, 1 eq.), the mixture was stirred at 25°C for 5 hours. The mixture was diluted with water (300 ml) and extracted with ethyl acetate (300 ml, twice), the organic phase was dried with anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the crude product as a brown oil. This crude product was dissolved in tetrahydrofuran/water (2/1, 150 ml) and lithium hydroxide (3 g) was added and the mixture was stirred at 25°C for 1 hour. The mixture was diluted with water (300 ml) and adjusted to pH=7 with aqueous hydrochloric acid (0.5 M), the mixture was extracted with ethyl acetate (300 ml, twice), the organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. This crude product was purified by silica gel chromatography (2-10% metonal in dichloromethane) to give tert-butyl (2S,4R)-4-hydroxy-2-[[2-hydroxy-4-(4-methylthiazol-5-yl )phenyl]methylcarbamoyl]pyrrolidine-1-carboxylate (6.9 g, 15.92 mmol, 50% yield) as a yellow oil. LC/MS (ESI) m/z: 434.1 [M+1] ⁺ .

[00700] Step 8: Preparation of (2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[00701] To a solution of tert-butyl (2S,4R)-4-hydroxy-2-[[2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidine-1-carboxylate (6.9 g, 15.92 mmol, 1 eq.) in methanol (30 ml) hydrochloride/dioxane (4 M, 30 ml, 7.54 eq.) was added and the mixture was stirred at 25°C for 20 minutes. The mixture was concentrated to give the product as a yellow solid, this crude product was triturated with ethyl acetate and petroleum ether (1:1, 20 ml), the mixture was filtered and the solid was dried on a rotary evaporator to give the product (2S,4R)-4- hydroxy-N-[[2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (4.83 g, 13.06 mmol, 82% yield, hydrochloric acid) as yellow solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.03 (br s, 1H), 9.11- 8.95 (m, 2H), 8.66 (br s, 1H), 7.20 (d, J=7.9 Hz, 1H), 7.04 (d, J=1.3 Hz, 1H), 6.90 (dd, J=1.7, 7.8 Hz, 1H), 4 .44 (br s, 1H), 4.40-4.26 (m, 3H), 3.41-3.27 (m, 1H), 3.13-3.02 (m, 1H), 2, 46 (s, 3H), 2.33 (br dd, J=7.5, 12.7 Hz, 1H), 1.96-1.85 (m, 1H), 1.33-1.24 (m , 1H).

[00702] Step 9: Preparation of (2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)-1-(3-methyl-2-(3- methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide

[00703] To a solution of (2S,4R)-4-hydroxy-N-[[2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (4.83 g, 13 .06 mmol, 1 eq., hydrochloride) in dimethylformamide (60 ml), diisopropylethylamine (5.06 g, 39.18 mmol, 6.82 ml, 3 eq.) was added, followed by 3-methyl-2-(3- methylisoxazol-5-yl)butanoic acid (2.39 g, 13.06 mmol, 1 eq.) and HATU (5.46 g, 14.36 mmol, 1.1 eq.), the mixture was stirred at 25°C in within 2 hours. The mixture was diluted with water (200 ml) and extracted with ethyl acetate (300 ml, twice), the organic phase was dried with anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the crude product. This crude product was purified by preparative HPLC, the acetonitrile fraction was removed and the residue was extracted with dichloromethane (300 ml x 5), the organic phase was dried with anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the product (2S,4R)-4-hydroxy-N-[ [2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]methyl]-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidin-2-carboxamide (4.0 g, 8.02 mmol, 61% yield) as a white solid. ¹ H-NMR (400 MHz, CD ₃ OD) ₃ 8.85 (s, 1H), 7.39-7.23 (m, 1H), 6.98-6.86 (m, 2H), 6, 31-6.06 (m, 1H), 4.65-4.28 (m, 4H), 3.94-3.48 (m, 3H), 2.52-2.45 (m, 3H), 2.42-2.31 (m, 1H), 2.26-2.15 (m, 4H), 2.13-2.03 (m, 1H), 1.08-1.01 (m, 3H ), 0.92-0.81 (m, 3H).

[00704] Example of the synthesis of 2-(2-((tert-butoxycarbonyl)(methyl)amino)ethoxy)ethyl 4-methylbenzenesulfonate

[00705] Step 1: Preparation of 2-(2-hydroxyethoxy)ethyl-4-methylbenzenesulfonate

[00706] Sodium hydride (6.29 g, 157, 27 mmol, 60% purity, 0.6 equiv) at 0°C and stirred for 0.5 hour under nitrogen atmosphere. Then p-toluenesulfonyl chloride (50 g, 262.26 mmol, 1 eq.) was added to the mixture, heated to 25° C. and stirred for 6 hours. The mixture was poured into saturated ammonium chloride solution (200 ml) and stirred for 15 minutes. The aqueous phase was extracted with ethyl acetate (200 ml x 2). The combined organic phase was washed with brine (200 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (petroleum ether/ethyl acetate 10/1 to 1/1) to give 2-(2-hydroxyethoxy)ethyl-4-methylbenzenesulfonate (35 g, 134.46 mmol, 51% yield) in in the form of yellow oil. LC/MS (ESI) m/z: 261.0 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.76-7.72 (m, 2H), 7.28 (d, J=8.0, 2H), 4.15-4.02 (m, 2H), 3.66-3.55 (m, 4H), 3.49-3.44 (m, 2H), 2.38 (s, 3H).

[00707] Step 2: Preparation of 2-(2-(methylamino)ethoxy)ethan-1-ol

[00708] A mixture of 2-(2-hydroxyethoxy)ethyl-4-methylbenzenesulfonate (3 g, 11.52 mmol, 1 eq.) in methamine (11.93 g, 115.25 mmol, 10 eq.) was degassed and purged with nitrogen 3 times, and then the mixture was stirred at 85°C for 16 hours under a nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure to obtain a residue. The compound 2-[2-(methylamino)ethoxy]ethanol (1.37 g, 11.50 mmol, 99% yield) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 3.72-3.47 (m, 8H), 2.67-2.63 (m, 3H).

[00709] Step 3: Preparation of tert-butyl (2-(2-hydroxyethoxy)ethyl)(methyl)carbamate

[00710] A mixture of 2-[2-(methylamino)ethoxy]ethanol (1.37 g, 11.50 mmol, 1 eq.) and di-tert-butyl dicarbonate (3.01 g, 13.80 mmol, 3.17 ml, 1.2 eq.) in dichloromethane (20 ml) was degassed and purged with nitrogen 3 times, and then the mixture was stirred at 25°C for 16 hours under a nitrogen atmosphere. The reaction mixture was quenched by adding 200 ml of water and extracted with ethyl acetate (300 ml x 3). The combined organic layers were washed with brine (100 ml x 3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (petroleum ether/ethyl acetate 50/1 to 5/1). The compound tert-butyl N-[2-(2-hydroxyethoxy)ethyl]-N-methylcarbamate (1.7 g, 7.75 mmol, 67% yield) was obtained as a yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 3.77-3.68 (m, 2H), 3.66-3.55 (m, 4H), 3.42 (br s, 2H), 2, 92 (s, 3H), 1.48-1.25 (m, 9H).

[00711] Step 4: Preparation of 2-(2-((tert-butoxycarbonyl)(methyl)amino)ethoxy)ethyl 4-methylbenzenesulfonate

[00712] A mixture of tert-butyl-N-[2-(2-hydroxyethoxy)ethyl]-N-methylcarbamate (1.1 g, 5.02 mmol, 1 eq.), p-toluenesulfonyl chloride (1.91 g, 10 .03 mmol, 2 equiv) and triethylamine (1.52 g, 15.05 mmol, 2.09 ml, 3 equiv) in dichloromethane (20 ml) were degassed and purged with nitrogen 3 times, and then the mixture was stirred at 25°C for 12 hours in a nitrogen atmosphere. The reaction mixture was quenched by adding 200 ml of water and extracted with ethyl acetate (300 ml x 3). The combined organic layers were washed with brine (100 ml x 3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (petroleum ether/ethyl acetate 30/1 to 10/1). The compound 2-[2-[tert-butoxycarbonyl(methyl)amino]ethoxy]ethyl-4-methylbenzenesulfonate (1.6 g, 4.28 mmol, 85% yield) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.73 (d, J=8.4 Hz, 2H), 7.28 (d, J=8.0 Hz, 2H), 4.12-4, 01 (m, 2H), 3.60-3.52 (m, 2H), 3.45 (br s, 2H), 3.25 (br s, 2H), 2.78 (s, 3H), 2 .38 (s, 3H), 1.37 (s, 9H).

[00713] Example of the synthesis of tert-butyl (2-(2-(2-(((2S,4R)-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl )butanoyl)pyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)ethyl)(methyl)carbamate and tert-butyl (2-(2-(2-(((2S ,4R)-4-hydroxy-1-((S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5- yl)phenoxy)ethoxy)ethyl)(methyl)carbamate

[00714] Step 1: Preparation of tert-butyl (2-(2-(2-(((2S,4R)-4-hydroxy-1-(3-methyl-2-(3-methylisoxazol-5-yl)butanoyl )pyrrolidin-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)ethyl)(methyl)carbamate

[00715] Mixture of 2-[2-[tert-butoxycarbonyl(methyl)amino]ethoxy]ethyl4-methylbenzenesulfonate (70 mg, 0.18 mmol, 1 eq.), (2S,4R)-4-hydroxy-N-[ [2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]methyl]-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidin-2-carboxamide (93 mg, 0.18 mmol, 1 eq.) and potassium carbonate (51 mg, 0.37 mmol, 2 eq.) in N,N-dimethylformamide (2 ml) were degassed and purged with nitrogen 3 times, and then the mixture was stirred at 80°C for 4 hours in a nitrogen atmosphere. The reaction mixture was quenched by adding water (10 ml) and extracted with ethyl acetate (20 ml x 3). The combined organic layers were washed with brine (10 ml (3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (9% methanol in dichloromethane) to give the compound tert-butyl N-[ 2-[2-[2-[[[(2S,4R)-4-hydroxy-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidin-2-carbonyl]amino]methyl ]-5-(4-methylthiazol-5-yl)phenoxy]ethoxy]ethyl]-N-methylcarbamate (100 mg, 0.14 mmol, 76% yield) as a yellow oil. LC/MS (ESI) m/z : 722.4 [M+23] ⁺ .

[00716] Step 2: Preparation of tert-butyl (2-(2-(2-(((2S,4R)-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazole-5 -yl)butanoyl)pyrrolidin-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)ethyl)(methyl)carbamate and tert-butyl (2-(2-(2-(( (2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol- 5-yl)phenoxy)ethoxy)ethyl)(methyl)carbamate

[00717] Tert-butyl N-[2-[2-[2-[[[(2S,4R)-4-hydroxy-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl] pyrrolidine-2-carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]ethoxy]ethyl]-N-methylcarbamate (250 mg, 0.35 mmol, 1 eq.) was purified by preparative SFC ( supercritical fluid chromatography, supercritical fluid chromatography). The compounds tert-butyl (2-(2-(2-(((2S,4R)-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)) were obtained pyrrolidine)-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)ethyl)(methyl)carbamate (90 mg, 0.12 mmol, 33% yield, 94% purity) and tert -butyl (2-(2-(2-(((2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidin-2 -carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)ethyl)(methyl)carbamate (90 mg, 0.12 mmol, 35% yield, 99% purity) as colorless oils.

[00718] Example of synthesis of methyl 2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoate

[00719] Step 1: Preparation of methyl 3-(benzyloxy)isoxazole-5-carboxylate

[00720] To a solution of methyl 3-hydroxyisoxazole-5-carboxylate (7.20 g, 50.31 mmol, 1.00 eq) in acetone (150 mL) was added potassium carbonate (13.91 g, 100.62 mmol , 2.00 eq.). The mixture was heated to 80°C for 1 hour, then (bromomethyl)benzene (10.33 g, 60.37 mmol, 1.20 eq.) was added. The resulting mixture was stirred at 80°C for another 3 hours. The solid was filtered and the filtrate was concentrated in vacuo. The residue was further purified by silica gel column chromatography (petroleum ether/ethyl acetate 15:1 to 10:1) to give methyl 3-benzyloxyisoxazole-5-carboxylate (9.50 g, 40.73 mmol, 81% yield) in the form of a colorless oil. The oil hardened after being kept at 15°C for 15 hours. LC/MS (ESI) m/z: 256.0 [M+23] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.49-7.41 (m, 5H), 6.60 (s, 1H), 5.34 (s, 2H), 3.97 (s, 3H ).

[00721] Step 2: Preparation of (3-(benzyloxy)isoxazol-5-yl)methanol

[00722] Sodium borohydride (756 mg, 19.98 mmol, 2 .00 eq.). The resulting mixture was stirred at a temperature of 15°C for 3 hours. The mixture was poured into hydrochloric acid (0.2 M, 200 ml) and then extracted with ethyl acetate (150 ml x 2). The combined organic layers were washed with brine (200 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give (3-benzyloxyisoxazol-5-yl)methanol (1.85 g, 9.02 mmol, 90% yield %) in the form of a colorless oil. LC/MS (ESI) m/z: 206.0 [M+1] ⁺ .

[00723] Step 3: Preparation of 2-(3-(benzyloxy)isoxazol-5-yl)acetonitrile

[00724] To a solution of cyanide bromide (334 mg, 3.15 mmol, 1.05 eq.) and triphenylphosphine (787 mg, 3.00 mmol, 1.00 eq.) in dichloromethane (10 ml) was added a solution of (3- benzyloxyisoxazol-5-yl)methanol (616 mg, 3.00 mmol, 1.00 eq.) in dichloromethane (10 ml). The mixture was stirred at 15°C for 1 hour, then 2,3,4,6,7,8,9,10-octahydropyrimido[1,2-a]azepine (480 mg, 3.15 mmol, 1.05 eq.) at 0°C. The resulting mixture was stirred at a temperature of 0-15°C for another 14 hours. The solvent was concentrated in vacuo. The residue was further purified by silica gel column chromatography (petroleum ether:ethyl acetate 5:1 to 4:1) to give 2-(3-benzyloxyisoxazol-5-yl)acetonitrile (320 mg, 1.49 mmol, 50% yield ) in the form of a colorless oil. LC/MS (ESI) m/z: 215.0 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.48-7.41 (m, 5H), 6.06 (s, 1H), 5.30 (s, 2H), 3.82 (s, 2H ).

[00725] Step 4: Preparation of 2-(3-(benzyloxy)isoxazol-5-yl)-3-methylbutanenitrile

[00726] Potassium carbonate (138 mg, 1 .00 mmol, 1.00 eq.). The mixture was stirred at 15°C for half an hour, then 2-iodopropane (170 mg, 1.00 mmol, 1.00 eq.) was added. The resulting mixture was stirred at 15°C for another 2.5 hours. Potassium 2-methylpropan-2-olate (90 mg, 0.8 mmol, 0.80 eq.) was then added to the mixture and the mixture was stirred at 15° C. for another 12 hours. The mixture was poured into hydrochloric acid (0.2 M, 30 ml) and then extracted with ethyl acetate (30 ml x 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography (10:1 to 8:1 petroleum ether:ethyl acetate) to give 2-(3-benzyloxyisoxazol-5-yl)-3-methylbutanenitrile (150 mg, 0.56 mmol, yield 59%) in the form of a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.48-7.41 (m, 5H), 6.04 (s, 1H), 5.28 (s, 2H), 3.85 (d, J =5.6 Hz, 1H), 2.42-2.37 (m, 1H), 1.18 (d, J=6.8 Hz, 3H), 1.10 (d, J=6.8 Hz , 3H).

[00727] Step 5: Preparation of 2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoic acid

[00728] To a solution of 2-(3-benzyloxyisoxazol-5-yl)-3-methylbutanenitrile (3.40 g, 13.27 mmol, 1.00 eq.) in dioxane (30 ml) was added hydrochloric acid (11.8 M, 120 ml). The mixture was heated to a temperature of 100°C and stirred at 100°C for 15 hours. The mixture was cooled to 15°C and then extracted with ethyl acetate (150 ml x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was further purified by preparative HPLC to give 2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoic acid (230 mg, 1.19 mmol, 9% yield) as a yellow solid. LC/MS (ESI) m/z: 186.1 [M+1] ⁺ .

[00729] Step 6: Preparation of methyl 2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoate

[00730] To a solution of 2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoic acid (1 g, 5.40 mmol, 1 eq.) in methanol (10 ml) was added thionyl chloride (2.57 g, 21% mmol, 1.57 ml, 4 eq.) at 0°C. The reaction mixture was stirred at 70°C for 3 hours. The reaction mixture was concentrated under reduced pressure. The residue was diluted with water (50 ml) and extracted with ethyl acetate (30 ml x 3). The combined organic layers were washed with brine (80 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Methyl 2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoate (1 g, 5.02 mmol, 92% yield) was obtained as a yellow oil. LC/MS (ESI) m/z: 200.1 [M+1] ⁺ .

[00731] Example of the synthesis of (2S,4R)-4-hydroxy-N-((S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide hydrochloride .

[00732] Step 1: Preparation of 1-(4-bromo-2-methoxyphenyl)ethan-1-one

[00733] To a solution of 1-(4-bromo-2-hydroxyphenyl)ethanone (10 g, 46.50 mmol, 1.0 eq.) in dimethylformamide (50 ml) was added potassium carbonate (9.64 g, 69.75 mmol, 1.5 equiv). Iodomethane (13.20 g, 93.00 mmol, 2.0 eq.) was then added to the mixture at 0°C. The mixture was then stirred at 20°C for 12 hours. The mixture was diluted with water (200 ml), extracted with ethyl acetate (100 ml x 3), washed with brine (50 ml x 2), dried over anhydrous sodium sulfate, filtered and then concentrated. 1-(4-bromo-2-methoxyphenyl)ethanone (10.5 g, 45.84 mmol, 98% yield) was obtained as a yellow solid. LC/MS (ESI) m/z: 230.9 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.63 (d, J=8.4 Hz, 1H), 7.18-7.11 (m, 2H), 3.93 (s, 3H), 2.60 (s, 3H).

[00734] Step 2: Preparation of (R,E)-N-(1-(4-bromo-2-methoxyphenyl)ethylidene)-2-methylpropane-2-sulfinamide.

[00735] To a solution of 1-(4-bromo-2-methoxyphenyl)ethanone (10 g, 43.65 mmol, 1.18 eq.) in tetrahydrofuran (50 ml) was added tetraethyl titanate (16.94 g, 74.26 mmol , 2.0 eq.). The mixture was then added with 2-methylpropane-2-sulfinamide (4.5 g, 37.13 mmol, 1.0 eq) and purged with N ₂ (3 times). The mixture was then stirred at 70°C for 12 hours. The mixture was quenched with water (100 ml), diluted with water (200 ml), filtered and then extracted with ethyl acetate (200 ml (3), washed with brine (200 ml), dried over anhydrous sodium sulfate, filtered and then concentrated. The residue was purified by column chromatography on silica (petroleum ether: ethyl acetate in the ratio from 20:1 to 3:1) to obtain (NE)-N-[1-(4-bromo-2-methoxyphenyl)ethylidene]-2-methylpropane-2-sulfinamide (9 g, 27.09 mmol, 73% yield) as a yellow oil ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.35 (d, J=8.0 Hz, 1H), 7.13 ( dd,.7=1.2, 8.0 Hz, 1H), 7.09 (s, 1H), 3.88 (s, 3H), 2.70 (s, 3H), 1.35-1, 28 (m, 9H).

[00736] Step 3: Preparation of (R)-N-((S)-1-(4-bromo-2-methoxyphenyl)ethyl)-2-methylpropane-2-sulfinamide

[00737] To a solution of (NE)-N-[1-(4-bromo-2-methoxyphenyl)ethylidene]-2-methylpropane-2-sulfinamide (9 g, 27.09 mmol, 1.0 eq.) in tetrahydrofuran (90 ml) L-celecride (1 M, 81.26 ml, 3.0 eq.) was added at 0°C. The mixture was then stirred at 20°C for 2 hours. The mixture was quenched with water (100 ml), diluted with water (20 ml), extracted with ethyl acetate (300 ml x 3), washed with brine (200 ml), dried over anhydrous sodium sulfate, filtered and then concentrated. The residue was purified by silica column chromatography (petroleum ether:ethyl acetate 20:1 to 1:1). N-[(1S)-1-(4-bromo-2-methoxyphenyl)ethyl]-2-methylpropane-2-sulfinamide (5.5 g, 16.45 mmol, 60% yield) was obtained as a yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.20-7.16 (m, 1H), 7.12-7.07 (m, 1H), 7.02 (d, 7=2.0 Hz , 1H), 4.90-4.82 (m, 1H), 3.86 (s, 3H), 3.50 (d, 7=5.2 Hz, 1H), 1.52 (d, 7= 6.8 Hz, 3H), 1.21 (s, 9H).

[00738] Step 4: Preparation of (R)-N-((S)-1-(2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboro lan-2-yl )phenyl)ethyl)-2-methylpropane-2-sulfinamide

[00739] To a solution of N-[(1S)-1-(4-bromo-2-methoxyphenyl)ethyl]-2-methylpropane-2-sulfinamide (4.7 g, 14.06 mmol, 1.0 eq.) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (5.36 g, 21.09 mmol, 1.5 eq.) in dioxane (12 ml) was added [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(ii) (1.03 g, 1.41 mmol, 0.1 eq. ) and potassium acetate (2.76 g, 28.12 mmol, 2.0 eq.). The mixture was then purged with N ₂ (3 times). The mixture was then stirred at 90°C for 5 hours. The mixture was diluted with water (20 ml), filtered and then extracted with ethyl acetate (50 ml x 3), washed with brine (30 ml), dried over anhydrous sodium sulfate, filtered and then concentrated. The residue was purified by silica column chromatography (petroleum ether:ethyl acetate 10:1 to 1:1). N-[(1S)-1-[2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]ethyl]-2-methylpropane-2 was obtained -sulfinamide (4.5 g, 11.80 mmol, 84% yield) as a yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.42 (d, J=8.0 Hz, 1H), 7.32-7.28 (m, 2H), 4.90 (m, J=6 .4 Hz, 1H), 3.91 (s, 3H), 3.73-3.67 (m, 1H), 1.55 (d, J=6.8 Hz, 3H), 1.36 (s , 12H), 1.19 (s, 9H).

[00740] Step 5: Preparation of (R)-N-((S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)-2-methylpropane-2-sulfinamide

[00741] To a solution of N-[(1S)-1-[2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]ethyl]-2 -methylpropane-2-sulfinamide (4.71 g, 12.36 mmol, 1.0 eq.) and 5-bromo-4-methylthiazole (2.2 g, 12.36 mmol, 1.0 eq.) in dioxane (8 ml) and water (2 ml) were added sodium bicarbonate (2.08 g, 24.72 mmol, 2.0 eq.) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (ii) (904 mg , 1.24 mmol, 0.1 eq.). The mixture was then purged with N ₂ (3 times). The mixture was then stirred at 90°C for 5 hours. The mixture was diluted with water (20 ml), filtered, extracted with ethyl acetate (30 ml x 3), washed with brine (30 ml), dried over anhydrous sodium sulfate, filtered and then concentrated. The residue was purified by silica column chromatography (dichloromethane:methanol = 100:1 to 10:1). N-[(1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-2-methylpropane-2-sulfinamide (3.9 g, 11.06 mmol, yield 89%) as a light yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.69 (s, 1H), 7.34 (d, J=8.0 Hz, 1H), 7.03 (d, J=8.0 Hz, 1H), 6.92 (s, 1H), 4.92 (m, J=6.4 Hz, 1H), 3.89 (s, 3H), 3.58 (d, J=5.6 Hz, 1H), 2.56 (s, 3H), 1.57 (d, J=6.8 Hz, 3H), 1.22 (s, 9H).

[00742] Step 6: Preparation of (S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethan-1-amine hydrochloride

[00743] To a solution of N-[(1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-2-methylpropane-2-sulfinamide (3.6 g, 10. 21 mmol, 1.0 eq.) in dichloromethane (20 mL), hydrochloric acid/dioxane (4 M, 18.46 mL, 7.23 eq.) was added. The mixture was then stirred at 20°C for 1 hour. The mixture was then concentrated. (1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethanamine (3.8 g, HCl) was obtained as a yellow solid.

[00744] Step 7: Preparation of tert-butyl (2S,4R.)-4-hydroxy-2-(((S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl )carbamoyl)pyrrolidine-1-carboxylate

[00745] To a solution of (1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethanamine (800 mg, 2.25 mmol, 1.0 eq., HCl) in dimethylformamide ( 10 mL), diisopropylethylamine (871 mg, 6.74 mmol, 1.17 mL, 3.0 eq.) was added. The mixture was then stirred at 20°C for 10 minutes. (2S,4R)-1-tert-butoxycarbonyl-4-hydroxypyrrolidine-2-carboxylic acid (519 mg, 2.25 mmol, 1.0 eq) and 1-hydroxybenzotriazole (364 mg, 2.70 mmol) were added to the mixture , 1.2 eq.) and stirred at 20°C for 10 minutes. 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (517 mg, 2.70 mmol, 1.2 eq.) was then added to the mixture and stirred at 20° C. for 40 minutes. The mixture was diluted with water (30 ml), extracted with ethyl acetate (30 ml x 3), washed with brine (30 ml), dried over anhydrous sodium sulfate, filtered and then concentrated. The residue was purified by silica column chromatography (dichloromethane:methanol = 100:1 to 20:1). We obtained tert-butyl (2S,4R)-4-hydroxy-2-[[(1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1- carboxylate (850 mg, 1.84 mmol, 82% yield) as a yellow solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.69 (s, 1H), 7.28 (s, 1H), 7.00 (dd, J=1.6, 7.6 Hz, 1H), 6.93 (s, 1H), 5.34-5.21 (m, 1H), 4.59-4.26 (m, 2H), 3.91 (s, 3H), 3.81-3, 53 (m, 2H), 2.54 (s, 3H), 2.51-2.38 (m, 1H), 2.22-1.94 (m, 1H), 1.57-1.18 ( m, 13H).

[00746] Step 8: Preparation of (2S,4R)-4-hydroxy-N-((S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2- carboxamide hydrochloride.

[00747] To a solution of tert-butyl (2S,4R)-4-hydroxy-2-[[(1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl] pyrrolidine-1-carboxylate (850 mg, 1.84 mmol, 1.0 eq.) in dichloromethane (10 ml) was added hydrochloric acid (gas)/dioxane (4 M, 10 ml). The mixture was then stirred at 20°C for 1 hour. The mixture was then concentrated. (2S,4R)-4-hydroxy-N-[(1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (700 mg, 1 .76 mmol, 95% yield, HCl) as a yellow solid.

[00748] Example of the synthesis of tert-butyl (2-(2-(2-((5-((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-( 2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)oxy)ethoxy) ethoxy)ethyl)(methyl)carbamate and tert-butyl (2-(2-(2-((5-((R)-1-((2S,4R)-4-hydroxy-2-(((S) -1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl) oxy)ethoxy)ethoxy)ethyl)(methyl)carbamate

[00749] Step 1: Preparation of methyl-3-methyl-2-(3-((2,2,5-trimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl)oxy) isoxazol-5-yl)butanoate

[00750] To a solution of methyl 2-(3-hydroxyisoxazol-5-yl)-3-methylbutanoate (500 mg, 2.51 mmol, 1.0 eq.) and 2-[2-[2-[tert-butoxycarbonyl (methyl)amino]ethoxy]ethoxy]ethyl-4-methylbenzenesulfonate (1.05 g, 2.51 mmol, 1.0 eq.) in dimethylformamide (10 ml) was added potassium carbonate (1.04 g, 7.53 mmol , 3.0 eq.). The mixture was then stirred at 70°C for 48 hours. The mixture was diluted with water (50 ml), extracted with ethyl acetate (50 ml x 3), washed with brine (50 ml), dried over anhydrous sodium sulfate, filtered and then concentrated. The residue was purified by silica column chromatography (petroleum ether:ethyl acetate 10:1 to 1:1). Methyl 2-[3-[2-[2-[2-[tert-butoxycarbonyl(methyl)amino]ethoxy]ethoxy]ethoxy]isoxazol-5-yl]-3-methylbutanoate (650 mg, 1.46 mmol, yield 58%) as a yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 5.93 (s, 1H), 4.44-4.37 (m, 2H), 3.88-3.83 (m, 2H), 3.74 (s, 3H), 3.72-3.60 (m, 6H), 3.50 (d, J=8.8 Hz, 1H), 3.41 (br s, 2H), 2.92 (s , 3H), 2.37 (m, J=6.8, 8.8 Hz, 1H), 1.47 (s, 9H), 1.01 (d, J=6.8 Hz, 3H), 0 .93 (d, J=6.8 Hz, 3H).

[00751] Step 2: Preparation of 3-methyl-2-(3-((2,2,5-trimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl)oxy)isoxazol- 5-yl)butanoic acid

[00752] To a solution of methyl 2-[3-[2-[2-[2-[tert-butoxycarbonyl(methyl)amino]ethoxy]ethoxy]ethoxy]isoxazol-5-yl]-3-methylbutanoate (630 mg, 1.42 mmol, 1.0 eq.) in methanol (4 mL) and water (2 mL), lithium hydroxide monohydrate (594 mg, 14.17 mmol, 10.0 eq.) was added. The mixture was then stirred at 20°C for 2 hours. The mixture was quenched with aqueous hydrochloric acid (4 M, 3 ml), extracted with ethyl acetate (30 ml x 3), washed with brine (20 ml x 2), dried over anhydrous sodium sulfate, filtered and then concentrated. This gave 2-[3-[2-[2-[2-[tert-butoxycarbonyl(methyl)amino]ethoxy]ethoxy]ethoxy]isoxazol-5-yl]-3-methylbutanoic acid (600 mg, 1.39 mmol, yield 98%) as a yellow oil.

[00753] Step 3: Preparation of tert-butyl (2-(2-(2-((5-(1-((2S,4R)-4-hydroxy-2-(((S)-1-(2- methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)oxy)ethoxy)ethoxy) ethyl)(methyl)carbamate

[00754] To a solution of (2S,4R)-4-hydroxy-N-[(1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide ( 369 mg, 0.93 mmol, 1.0 eq., HCl) in dimethylformamide (5 ml) was added diisopropylethylamine (360 mg, 2.79 mmol, 3.0 eq.). The mixture was then stirred at 20°C for 10 minutes. 2-[3-[2-[2-[2-[tert-Butoxycarbonyl(methyl)amino]ethoxy]ethoxy]ethoxy]isoxazol-5-yl]-3-methylbutanoic acid (400 mg, 0.93 mmol, 1.0 eq) and 1-hydroxybenzotriazole (251 mg, 1.86 mmol, 2.0 eq) and stirred at 20°C for 10 minutes. 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (356 mg, 1.86 mmol, 2.0 eq.) was then added to the mixture and stirred at 20° C. for 40 minutes. The mixture was diluted with water (30 ml), extracted with ethyl acetate (30 ml×2), washed with brine (30 ml), dried over anhydrous sodium sulfate, filtered and then concentrated. The residue was purified by preparative thin layer chromatography (dichloromethane:methanol = 10:1). We obtained tert-butyl N-[2-[2-[2-[5-[1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[2-methoxy-4-( 4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2-methylpropyl]isoxazol-3-yl]oxyethoxy]ethoxy]ethyl]-N-methylcarbamate (490 mg, 0.63 mmol, yield 67%, purity 99%) as a white solid.

[00755] Step 4: Preparation of tert-butyl (2-(2-(2-((5-((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1 -(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)oxy) ethoxy)ethoxy)ethyl)(methyl)carbamate and tert-butyl(2-(2-(2-((5-((R)-1-((2S,4R)-4-hydroxy-2-((( S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3- yl)oxy)ethoxy)ethoxy)ethyl)(methyl)carbamate

[00756] Tert-butyl N-[2-[2-[2-[5-[1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[2-methoxy-4 -(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2-methylpropyl]isoxazol-3-yl]oxyethoxy]ethoxy]ethyl]-N-methylcarbamate (490 mg, 0.63 mmol, 1.0 eq.) were separated by SFC. The organic layers were concentrated. We obtained tert-butyl(2-(2-(2-((5-((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(2-methoxy- 4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)oxy)ethoxy)ethoxy)ethyl) (methyl)carbamate (190 mg, 0.24 mmol, 95% yield, 98% purity) as a white solid. We obtained tert-butyl (2-(2-(2-((5-((R)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(2-methoxy- 4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)oxy)ethoxy)ethoxy)ethyl) (methyl)carbamate (250 mg, 0.32 mmol, 84% yield, 99% purity) as a white solid.

[00757] Synthesis example of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene-2-ol

[00758] Step 1: Preparation of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene-2-ol

[00759] To a solution of 4-bromonaphthalene-2-ol (220 mg, 0.99 mmol, 1 eq.) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1 [1,1'-bis(diphenylphosphino) )ferrocene]dichloropalladium(II)dichloromethane (80 mg, 0.1 mmol, 0.1 eq.) and potassium acetate (242 mg, 2.47 mmol, 2.5 eq.), and then the mixture was stirred at 90°C in a nitrogen atmosphere for 5 hours. The mixture was diluted with water (50 ml) and extracted with ethyl acetate (50 ml), the organic phase was dried with anhydrous (compound), filtered and the filtrate was concentrated to obtain the crude product. This crude product was purified by silica gel column chromatography (10-33.3% ethyl acetate in petroleum ether) to give 400 mg of product as a white solid, this product was further purified by preparative TLC (50% ethyl acetate in petroleum ether) to give 4 -(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene-2-ol (180 mg, 0.67 mmol, 67% yield) as a white solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.67 (d, J=8.2 Hz, 1H), 7.72-7.66 (m, 2H), 7.46-7.35 (m , 2H), 7.28-7.27 (m, 1H), 4.95 (s, 1H), 1.43 (s, 12H).

[00760] Example of the synthesis of tert-butyl-4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-2-(((R)-1-oxopropan-2-yl)oxy) quinazolin-4-yl)piperazine-1-carboxylate

[00761] Step 1: Preparation of methyl (R)-2-(benzyloxy)propanoate

[00762] A solution of (2R)-2-benzyloxypropanoic acid (20.00 g, 110.99 mmol, 1.00 eq.) in methanol (150 ml) was cooled to 0° C., then sulfur dioxide (39, 61 g, 332.96 mmol, 24.2 ml, 3.00 eq.). The mixture was then stirred at 50°C for 4 hours. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was diluted with saturated sodium bicarbonate solution (200 ml), then extracted with ethyl acetate (200 ml x 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the product, methyl (2R)-2-benzyloxypropanoate (21.96 g) as a yellow oil. LC/MS (ESI) m/z: 217.1 [M+23] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.40-7.27 (m, 5H), 4.70 (d, J=11.6 Hz, 1H), 4.46 (d, J=11 .6 Hz, 1H), 4.08 (q, J=6.8 Hz, 1H), 3.77 (s, 3H), 1.45 (d, J=6.8 Hz, 3H).

[00763] Step 2: Preparation of (R)-2-(benzyloxy)propanal

[00764] A solution of methyl (2R)-2-benzyloxypropanoate (20.96 g, 107.92 mmol, 1.00 eq.) in dichloromethane (200 ml) was cooled to minus 78° C., then diisobutylaluminum hydride (1 M) was added dropwise , 110 ml, 1.00 eq.). The mixture was then stirred at minus 78°C for 1 hour. The reaction mixture was quenched with hydrochloric acid (1 M, 10 ml) and filtered through celite. The filtrate was diluted with water (100 ml), extracted with dichloromethane (100 ml×2). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The compound (2R)-2-benzyloxypropanal (15.70 g) was obtained as a colorless oil. LC/MS (ESI) m/z: 181.1 [M+17] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 9.68 (d, J=1.6 Hz, 1H), 7.42-7.33 (m, 5H), 4.68-4.60 (m, 2H), 3.95-3.86 (m, 1H) , 1.34 (d, J=6.8 Hz, 3H).

[00765] Step 3: Preparation of (R)-(((1,1-dimethoxypropan-2-yl)oxy)methyl)benzene

[00766] To a solution of (2R)-2-benzyloxypropanal (14.70 g, 89.52 mmol, 1 eq.) in trimethoxymethane (71.15 g, 670.46 mmol, 73.5 mL, 7.49 eq. ) 4-methylbenzenesulfonic acid was added; pyridine (450 mg, 1.79 mmol, 0.02 eq.). The mixture was stirred at 25°C for 2 hours. The reaction mixture was diluted with water (100 ml), then extracted with ethyl acetate (100 ml×2). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 25/1 to 20/1) to obtain the desired product. The compound [(1R)-2,2-dimethoxy-1-methylethoxy]methylbenzene (16.70 g, 79.42 mmol, 89% yield, 100% purity) was obtained as a colorless oil. LC/MS (ESI) m/z: 233.1 [M+23] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.39-7.27 (m, 5H), 4.68-4.61 (m, 2H), 4.22 (d, J=5.2 Hz , 1H), 3.62-3.53 (m, 1H), 3.44 (d, J=4.0 Hz, 6H), 1.20 (d, J=6.4 Hz, 3H).

[00767] Step 4: Preparation of (R)-1,1-dimethoxypropan-2-ol.

[00768] Palladium on activated carbon ( 500 mg, purity 5%) and palladium hydroxide (500 mg, purity 5%) under nitrogen atmosphere. The suspension was degassed in vacuum and purged several times with hydrogen gas. The mixture was stirred under an atmosphere of hydrogen gas (15 psi) at 60°C for 8 hours. The reaction mixture was filtered through celite and concentrated under reduced pressure to obtain a residue. The compound (2R)-1,1-dimethoxypropan-2-ol (4.1 g) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.08 (d, J=6.4 Hz, 1H), 3.81-3.73 (m, 1H), 3.45 (d, J=4.0 Hz, 6H), 1.20 (d, J=6.4 Hz, 3H).

[00769] Step 5: Preparation of tert-butyl (R)-4-(7-bromo-6-chloro-2-((1,1-dimethoxypropan-2-yl)oxy)-8-fluoroquinazolin-4-yl) piperazine-1-carboxylate

[00770] Tert-Butyl 4-(7-bromo-2,6-dichloro-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate (300 mg, 0.62 mmol, 1 eq.), (2R)- 1,1-dimethoxypropan-2-ol (150.13 mg, 1.25 mmol, 2 eq.), potassium carbonate (259 mg, 1.87 mmol, 3 eq.) and 1,4-diazabicyclo[2.2.2 ]octane (7 mg, 62.48 μmol, 0.1 eq.) was placed in a microwave-safe tube in acetonitrile (10 mL). The sealed tube was heated at 100°C for 2 hours in a microwave oven. The reaction mixture was filtered and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by silica gel column chromatography (0-15% ethyl acetate in petroleum ether) to give tert-butyl-4-[7-bromo-6-chloro-2-[(1R)-2,2-dimethoxy-1-methylethoxy ]-8-fluoroquinazolin-4-yl]piperazin-1-carboxylate (706 mg, 1.10 mmol, 22% yield, 87% purity) as a yellow solid. LC/MS (ESI) m/z: 565.1 [M+1] ⁺ .

[00771] Step 6: Preparation of tert-butyl-4-(6-chloro-2-(((R)-1,1-dimethoxypropan-2-yl)oxy)-8-fluoro-7-(3-hydroxynaphthalene- 1-yl)quinazolin-4-yl)piperazine-1-carboxylate

[00772] To a solution of tert-butyl-4-[7-bromo-6-chloro-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-8-fluoroquinazolin-4-yl]piperazin-1- carboxylate (608 mg, 1.08 mmol, 1 eq.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene-2-ol (379 mg 1, 40 mmol, 1.3 eq.) in tetrahydrofuran (15 ml), potassium phosphate (1.5 M, 2.16 ml, 3 eq.) and (2-dicyclohexylphosphino-2',4',6'-triisopropyl- 1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(ii) methanesulfonate (91 mg, 0.11 mmol, 0.1 eq.). The reaction mixture was degassed and charged with nitrogen 3 times and then heated to 65°C for 16 hours. Ethyl acetate (30 ml) was added and the mixture was washed with water (30 ml). The organic layer was dried over sodium sulfate and then concentrated in vacuo to obtain a residue. The residue was purified by flash silica gel chromatography (0-60% ethyl acetate in petroleum ether) to give the crude product (600 mg). This crude product was purified by semipreparative reverse phase HPLC. The collected fractions were concentrated in vacuo to remove most of the acetonitrile. The pH of the mixture was adjusted to 8 with saturated aqueous sodium bicarbonate and then extracted with ethyl acetate (50 ml×2). The combined organic layer was dried over sodium sulfate and then concentrated in vacuo to give tert-butyl-4-[6-chloro-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-8-fluoro-7- (3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (400 mg) as a light yellow solid. LC/MS (ESI) m/z: 627.2 [M+1] ⁺ .

[00773] Step 7: Preparation of (2R)-2-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl) oxy)propanal

[00774] To a solution of tert-butyl-4-[6-chloro-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline -4-yl]piperazine-1-carboxylate (400 mg, 0.64 mmol, 1 eq.) in dioxane (20 mL) was added hydrochloric acid (12 M, 2.00 mL, 37.63 eq.). The reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated in vacuo to give (2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazin-1-ylquinazolin-2-yl]oxypropanal (330 mg , hydrochloride) in the form of a light yellow gum. LC/MS (ESI) m/z: 481.1 [M+1] ⁺ .

[00775] Step 8: Preparation of tert-butyl-4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-2-(((R)-1-oxopropan-2-yl) hydroxy)quinazolin-4-yl)piperazine-1-carboxylate

[00776] Mixture of (2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxypropanal (330 mg, 0.64 mmol, 1 eq., hydrochloride) and di-tert-butyl dicarbonate (278.41 mg, 1.28 mmol, 2 eq.) in tetrahydrofuran (20 ml) were cooled to 0°C. Saturated aqueous sodium sulfate (322 mg, 3.83 mmol, 6 mL, 6 eq.) was then added. The reaction mixture was stirred at 25°C for 2 hours. Ethyl acetate (30 ml) and water (20 ml) were added and the mixture was separated. The organic layer was dried over sodium sulfate and then concentrated in vacuo to obtain a residue. The residue was purified by silica gel column chromatography with dichloromethane (50 ml) then ethyl acetate (40 ml) to give tert-butyl-4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2 -[(1R)-1-methyl-2-oxoethoxy]quinazolin-4-yl]piperazine-1-carboxylate (380 mg) as a light yellow solid. LC/MS (ESI) m/z: 581.2 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 9.71 (dd, J=1.8, 3.4 Hz, 1H), 7.80 (d, J=1.1 Hz, 1H), 7, 76 (d, J=8.3 Hz, 1H), 7.44 (dt, J=1.5, 7.3 Hz, 1H), 7.33-7.27 (m, 2H), 7.27 -7.22 (m, 1H), 7.10 (d, J=2.4 Hz, 1H), 6.32-5.88 (m, 1H), 5.30-5.22 (m, 1H ), 3.99-3.77 (m, 4H), 3.74-3.61 (m, 4H), 1.59-1.54 (m, 3H), 1.52 (s, 9H).

[00777] Example of the synthesis of tert-butyl 4-(2-(tosyloxy)ethoxy)piperidine-1-carboxylate

[00778] Step 1: Preparation of 2-(benzyloxy)ethyl-4-methylbenzenesulfonate

[00779] To a solution of 2-benzyloxyethanol (50 g, 328.54 mmol, 46.73 mL, 1 eq.) and potassium hydroxide (22.12 g, 394.24 mmol, 1.2 eq.) in tetrahydrofuran (200 ml) toluenesulfonyl chloride (56.37 g, 295.68 mmol, 0.9 eq.) was added. The mixture was stirred at 25°C for 1 hour. Ethyl acetate (1000 ml) was added, then the reaction mixture was filtered, then the filtrate was washed with brine (200 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (petroleum ether:ethyl acetate 5:1 to 3:1). The compound 2-benzyloxyethyl 4-methylbenzenesulfonate (75 g, 243.82 mmol, 74% yield, 99% purity) was obtained as a yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.85-7.79 (m, 2H), 7.38-7.31 (m, 4H), 7.30-7.26 (m, 2H) , 7.31 (s, 1H), 4.51 (s, 2H), 4.26-4.19 (m, 2H), 3.73-3.65 (m, 2H), 2.50-2 .40 (m, 3H).

[00780] Step 2: Preparation of tert-butyl 4-(2-(benzyloxy)ethoxy)piperidine-1-carboxylate

[00781] To a solution of tert-butyl-4-hydroxypiperidine-1-carboxylate (10.35 g, 51.41 mmol, 1.05 eq.) in N,N-dimethylformamide (150 mL) was added sodium cyanide (2.15 g, 53.86 mmol, 60% purity, 1.1 eq.) at 0°C, stirring for 0.5 hour, added 2-benzyloxyethyl-4-methylbenzenesulfonate (15 g, 48.96 mmol, 1 eq. ). The mixture was stirred at 25°C for 1 hour. Ammonium chloride solution (200 ml) was added and then the aqueous phase was extracted with ethyl acetate (200 ml*3). The combined organic phase was washed with brine (100 ml*2), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (petroleum ether:ethyl acetate 10:1 to 4:1). The compound tert-butyl 4-(2-benzyloxyethoxy)piperidine-1-carboxylate (14.5 g) was obtained as a colorless oil. LC/MS (ESI) m/z: 236.1 [M-100] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.41-7.30 (m, 5H), 4.60 (s, 2H), 3.80 (br d, J=9.9 Hz, 2H) , 3.71-3.62 (m, 4H), 3.55-3.46 (m, 1H), 3.08 (ddd, J=3.5, 9.5, 13.3 Hz, 2H) , 1.86 (br d, J=7.9 Hz, 2H), 1.60-1.50 (m, 2H), 1.47 (s, 9H).

[00782] Step 3: Preparation of tert-butyl 4-(2-hydroxyethoxy)piperidine-1-carboxylate

[00783] To a solution of tert-butyl-4-(2-benzyloxyethoxy)piperidine-1-carboxylate (5 g, 14.91 mmol, 1 eq.) in methanol (40 ml) was added palladium on activated carbon (0.5 g , purity 10%) in a nitrogen atmosphere. The suspension was degassed in vacuo and purged several times with H ₂ . The mixture was stirred under a hydrogen atmosphere (15 psi) at 25°C for 12 hours. The reaction mixture was filtered and concentrated in vacuo. The compound tert-butyl 4-(2-hydroxyethoxy)piperidine-1-carboxylate (3 g, 12.23 mmol, 82% yield) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 3.86-3.72 (m, 4H), 3.66-3.58 (m, 2H), 3.56-3.45 (m, 1H) , 3.16-3.07 (m, 2H), 2.15-2.05 (m, 1H), 1.96-1.78 (m, 2H), 1.59-1.49 (m, 2H), 1.47 (s, 9H).

[00784] Step 4: Preparation of tert-butyl 4-(2-(tosyloxy)ethoxy)piperidine-1-carboxylate

[00785] To a mixture of tert-butyl 4-(2-hydroxyethoxy)piperidine-1-carboxylate (7.4 g, 30.17 mmol, 1 eq.) and triethylamine (9.16 g, 90.50 mmol, 12. 60 ml, 3 eq.) in dichloromethane (70 ml) was added toluenesulfonyl chloride (8.63 g, 45.25 mmol, 1.5 eq.) at 0°C. The mixture was stirred at 25°C for 12 hours. The reaction mixture was concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (petroleum ether:ethyl acetate 4:1 to 1:1). The compound tert-butyl 4-[2-(p-tolylsulfonyloxy)ethoxy]piperidine-1-carboxylate (8.7 g, 21.78 mmol, 72% yield) was obtained as a yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.80 (d, J=8.3 Hz, 2H), 7.34 (d, J=8.1 Hz, 2H), 4.18-4, 13 (m, 2H), 3.76-3.70 (m, 2H), 3.67-3.60 (m, 4H), 3.47-3.39 (m, 1H), 3.14- 3.01 (m, 2H), 2.46-2.43 (m, 3H), 1.90-1.78 (m, 2H), 1.45 (s, 9H).

[00786] Example of the synthesis of tert-butyl 4-(2-(2-(2-(1,3-dioxyisoindolin-2-yl)ethoxy)ethoxy)ethoxy)piperidine-1-carbamate

[00787] Step 1: Preparation of 2-(2-(2-(benzyloxy)ethoxy)ethoxy)ethyl-4-methylbenzenesulfonate

[00788] A solution of 2-[2-(2-benzyloxyethoxy)ethoxy]ethanol (18.2 g, 75.74 mmol, 1 eq.) and potassium hydroxide (12.75 g, 227.22 mmol, 3 eq.) in tetrahydrofuran (100 ml) was stirred at 25°C for 0.5 hour, then p-toluenesulfonyl chloride (28.88 g, 151.48 mmol, 2 eq.) was added and stirred at 25°C for 1 hour. The reaction mixture was quenched by adding water (100 ml) at 25°C, and extracted with ethyl acetate (200 ml*3). The combined organic layers were washed with brine (150 ml*2), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash chromatography on silica gel (0-10% ethyl acetate in petroleum ether). The compound 2-[2-(2-benzyloxyethoxy)ethoxy]ethyl-4-methylbenzenesulfonate (20 g, 50.70 mmol, 66.9% yield) was obtained as a colorless oil.

[00789] Step 2: Preparation of tert-butyl 4-(2-(2-(2-(benzyloxy)ethoxy)ethoxy)ethoxy)piperidine-1-carboxylate

[00790] Sodium hydride (1.75 g, 43.73 mmol, 60% in mineral oil, 1.15 eq.) at 0°C. The mixture was then stirred at 25°C for 0.5 hours. 2-[2-(2-Benzyloxyethoxy)ethoxy]ethyl-4-methylbenzenesulfonate (15 g, 38.02 mmol, 1 eq.) was then added to the mixture. The mixture was stirred at 25°C for 2 hours. The mixture was quenched with saturated ammonium chloride solution (350 ml). The mixture was then extracted with ethyl acetate (150 ml×3). The combined organic layer was washed with water (200 ml×2) and brine (200 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate 10:1 to 3:1). The compound tert-butyl 4-[2-[2-(2-benzyloxyethoxy)ethoxy]ethoxy]piperidine-1-carboxylate (11.8 g, 27.86 mmol, 73.27% yield) was obtained as a colorless oil. LC/MS (ESI) m/z: 324.1 [M-99] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.35-7.28 (m, 5H), 4.57-4.56 (m, 2H), 3.84-3.75 (m, 2H) , 3.69-3.63 (m, 12H), 3.50-3.45 (m, 1H), 3.07-3.02 (m, 2H), 1.83-1.81 (m, 2H), 1.53-1.45 (m, 11H).

[00791] Step 3: Preparation of tert-butyl 4-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)piperidine-1-carboxylate

[00792] To a mixture of tert-butyl 4-[2-[2-(2-benzyloxyethoxy)ethoxy]ethoxy]piperidine-1-carboxylate (11.8 g, 27.86 mmol, 1 eq.) in methanol (100 ml) palladium on activated carbon catalyst (800 mg, 10% purity) was added. The mixture was degassed and charged with hydrogen 3 times. It was then stirred at 25°C for 12 hours under a hydrogen atmosphere (50 psi). The reaction mixture was heated to 60°C. The mixture was stirred at 60°C for an additional 12 hours under a hydrogen atmosphere (50 psi). The mixture was filtered. The filtrate was concentrated under reduced pressure to give the product, tert-butyl 4-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]piperidine-1-carboxylate (9 g) as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 3.82-3.60 (m, 14H), 3.50-3.47 (m, 1H), 3.06-3.02 (m, 2H) , 1.85-1.82 (m, 2H), 1.53-1.44 (m, 11H).

[00793] Step 4: Preparation of tert-butyl 4-(2-(2-(2-(tosyloxy)ethoxy)ethoxy)ethoxy)piperidine-1-carboxylate

[00794] To a mixture of tert-butyl-4-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]piperidine-1-carboxylate (7 g, 20.99 mmol, 1 eq.) and potassium hydroxide (1, 53 g, 27.29 mmol, 1.3 eq.) in tetrahydrofuran (70 ml) p-toluenesulfonyl chloride (4.80 g, 25.19 mmol, 1.2 eq.) was added. The mixture was stirred at 25°C for 1 hour. The mixture was diluted with ice water (100 ml). The mixture was then extracted with ethyl acetate (100 ml×3). The combined organic layer was washed with brine (150 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate 10:1 to 1:2). The compound obtained was tert-butyl 4-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (7.5 g, 15.38 mmol, 73.26% yield) in the form of a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.78 (d, J=8.4 Hz, 2H), 7.33 (d, J=8.4 Hz, 2H), 4.14 (t, J=4.8 Hz, 2H), 3.82-3.75 (m, 2H), 3.69-3.66 (m, 2H), 3.59-3.57 (m, 8H), 3 .45-3.35 (m, 1H), 3.05-3.00 (m, 2H), 2.43 (s, 3H), 1.81-1.80 (m, 2H), 1.50 -1.44 (m, 11H).

[00795] Step 5: Preparation of tert-butyl 4-(2-(2-(2-(1,3-dioxyisoindolin-2-yl)ethoxy)ethoxy)ethoxy)piperidine-1-carboxylate

[00796] To a mixture of tert-butyl 4-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (3.5 g, 7.18 mmol, 1 eq. ) in N,N-dimethylformamide (30 ml) was added potassium (1,3-dioxoisoindolin-2-yl) (1.60 g, 8.61 mmol, 1.2 eq). The mixture was stirred at 80°C for 2 hours. The mixture was diluted with water (100 ml). The mixture was then extracted with ethyl acetate (50 ml×3). The combined organic layer was washed with water (80 ml×2) and brine (80 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate 3:1 to 1:1). The compound obtained was tert-butyl 4-[2-[2-[2-(1,3-dioxoisoindolin-2-yl)ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (2.31 g, 4.99 mmol, yield 69%) in the form of a colorless oil. LC/MS (ESI) m/z: 363.1 [M+100] ⁺ .

[00797] Example of the synthesis of tert-butyl 4-(2-(2-(tosyloxy)ethoxy)ethoxy)piperidine-1-carboxylate

[00798] Step 1: Preparation of 2-(2-(benzyloxy)ethoxy)ethyl-4-methylbenzenesulfonate

[00799] To a solution of 2-(2-benzyloxyethoxy)ethanol (50 g, 254.79 mmol, 1 eq.) and potassium hydroxide (17.15 g, 305.74 mmol, 1.2 eq.) in tetrahydrofuran (200 ml) paratoluenesulfonyl chloride (48.57 g, 254.79 mmol, 1 eq.) was added. The mixture was stirred at 25°C for 1 hour. Ice water (500 ml) and ethyl acetate (500 ml) were added and the aqueous phase was extracted with ethyl acetate (300 ml*3). The combined organic phase was washed with brine (200 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (petroleum ether:ethyl acetate 5:1 to 3:1). The compound 2-(2-benzyloxyethoxy)ethyl-4-methylbenzenesulfonate (70 g, 199.76 mmol, 78% yield) was obtained as a yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.85-7.75 (m, 2H), 7.38-7.28 (m, 7H), 4.54 (s, 2H), 4.23 -4.13 (m, 2H), 3.72-3.69 (m, 2H), 3.64-3.60 (m, 2H), 3.59-3.55 (m, 2H), 2 .44 (s, 3H).

[00800] Step 2: Preparation of tert-butyl 4-(2-(2-(benzyloxy)ethoxy)ethoxy)piperidine-1-carboxylate

[00801] Sodium hydride (2.55 g, 63.75 mmol, 60% purity, 1.12 eq.) at 0°C, stirred for 0.5 hour. 2-(2-Benzyloxyethoxy)ethyl-4-methylbenzenesulfonate (20 g, 57.07 mmol, 1 eq.) was then added and the mixture was stirred at 25° C. for 1 hour. Ammonium chloride solution (200 ml) was added, then the aqueous phase was extracted with ethyl acetate (200 ml*3). The combined organic phase was washed with brine (100 ml*2), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (petroleum ether:ethyl acetate 5:1 to 3:1). The compound tert-butyl 4-[2-(2-benzyloxyethoxy)ethoxy]piperidine-1-carboxylate (16.2 g) was obtained as a yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.38-7.33 (m, 4H), 7.32-7.28 (m, 1H), 4.58 (s, 2H), 4.15 -4.08 (m, 1H), 3.77 (d, J=13.0 Hz, 2H), 3.72-3.66 (m, 2H), 3.66-3.63 (m, 5H ), 3.61-3.42 (m, 1H), 3.05 (ddd, J=3.4, 9.5, 13.3 Hz, 2H), 1.83 (d, J=7.1 Hz, 2H), 1.52 (tdd, J=4.5, 8.7, 13.0 Hz, 2H), 1.46 (s, 9H).

[00802] Step 3: Preparation of tert-butyl 4-(2-(2-hydroxyethoxy)ethoxy)piperidine-1-carboxylate

[00803] To a solution of tert-butyl-4-[2-(2-benzyloxyethoxy)ethoxy]piperidine-1-carboxylate (16.2 g, 42.69 mmol, 1 eq.) in methanol (100 ml) was added palladium carbon (1.5 g, 42.69 mmol, 4.27 ml, 10% purity, 1 eq.) under nitrogen. The suspension was degassed in vacuum and purged with hydrogen several times. The mixture was stirred under a hydrogen atmosphere (50 psi) at 25°C for 60 hours. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The compound tert-butyl 4-[2-(2-hydroxyethoxy)ethoxy]piperidine-1-carboxylate (6.7 g, 23.15 mmol, 54% yield) was obtained as a yellow oil. ¹ H-NMR (400 MHz, CD ₃ OD) δ 3.79-3.71 (m, 2H), 3.70-3.61 (m, 7H), 3.59-3.55 (m, 3H ), 3.21-3.06 (m, 2H), 1.91-1.81 (m, 2H), 1.51-1.42 (m, 12H).

[00804] Step 4: Preparation of tert-butyl 4-(2-(2-(tosyloxy)ethoxy)ethoxy)piperidine-1-carboxylate

[00805] To a solution of tert-butyl-4-[2-(2-hydroxyethoxy)ethoxy]piperidine-1-carboxylate (6.5 g, 22.46 mmol, 1 eq.) in dichloromethane (65 mL) was added triethylamine ( 6.82 g, 67.39 mmol, 9.38 ml, 3 eq.). p-Toluenesulfonyl chloride (6.42 g, 33.69 mmol, 1.5 eq.) was then added to the mixture. The mixture was stirred at 25°C for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography (0~25% ethyl acetate/petroleum ether). The compound tert-butyl 4-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]piperidine-1-carboxylate (7.4 g, 16.68 mmol, 74% yield) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CD ₃ OD) δ 7.81-7.76 (m, 2H), 7.46-7.41 (m, 2H), 4.81 (s, 3H), 3, 74-3.64 (m, 4H), 3.58-3.49 (m, 6H), 3.11 (t, J=9.6 Hz, 2H), 2.47-2.45 (m, 1H), 1.86-1.76 (m, 2H), 1.46 (s, 9H), 1.48-1.37 (m, 2H).

[00806] Synthesis example of N-(1,3-dimethyl-2-oxo-6-(3-propoxyphenoxy)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-4-hydroxy-N -((2-(trimethylsilyl)ethoxy)methyl)benzenesulfonamide

[00807] Step 1: Preparation of sodium 4-(benzyloxy)benzenesulfonate

[00808] To a solution of 4-hydroxybenzenesulfonic acid (17.50 g, 100.47 mmol, 1 eq.) in propan-2-ol (280 ml) was added (bromomethyl)benzene (36.86 g, 215.54 mmol, 25.6 ml, 2.15 eq.), sodium hydroxide (2 M, 100 ml, 1.99 eq.) and dropwise (bromomethyl)benzene (36.86 g, 215.54 mmol, 25.6 ml, 2.15 eq.). The mixture was stirred at 70°C for 20 hours. The mixture was concentrated to give a solid, the solid was washed with water (50 mL), filtered and concentrated under reduced pressure to give the product (4-benzyloxyphenyl)sulfonyloxysodium (19.00 g, 66.37 mmol, 66% yield) as a white solid substances. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.52 (d, J=8.8 Hz, 2H), 7.46-7.32 (m, 5H), 6.94 (d, J =8.8 Hz, 2H), 5.12 (s, 2H).

[00809] Step 2: Preparation of 4-(benzyloxy)benzenesulfonyl chloride

[00810] To a solution of (4-benzyloxyphenyl)sulfonyloxysodium (19.00 g, 66.37 mmol, 1.00 eq.) in sulfur dichloride (80 ml) was added N,N-dimethylformamide (475 mg, 6.50 mmol, 0.5 ml, 0.98 eq.). The mixture was stirred at 70°C for 4 hours. The mixture was concentrated to give the crude product and this material was dissolved in ethyl acetate (50 ml) and washed with water (150 ml) followed by three washes with saturated sodium bicarbonate and brine, filtered and the filtrate was condensed to give a white solid. The compound 4-benzyloxybenzenesulfonyl chloride (16.50 g, 58.36 mmol, 88% yield) was obtained as a white solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.00 (dd, J=2.0, 6.8 Hz, 2H), 7.46-7.40 (m, 5H), 7.15 (dd , J=2.0, 6.8 Hz, 2H), 5.21 (s, 2H).

[00811] Step 3: Preparation of 4-(benzyloxy)-N-(1,3-dimethyl-2-oxo-6-(3-propoxyphenoxy)-2,3-dihydro-1H-benzo[d]imidazole-5- yl)benzenesulfonamide

[00812] To a solution of 5-amino-1,3-dimethyl-6-(3-propoxyphenoxy)benzimidazol-2-one (3.00 g, 9.16 mmol, 1.00 eq.) and triethylamine (1.39 g, 13.75 mmol, 2.0 ml, 1.50 eq.) in dichloromethane (30 ml) add dropwise a solution of 4-benzyloxybenzenesulfonyl chloride (2.60 g, 9.16 mmol, 1.00 eq.) in dichloromethane (10 ml) at 0°C. The mixture was stirred at 20°C for 12 hours. Water (20 ml) was added to the mixture, then extracted with ethyl acetate (50 ml x 3), the organic layers were washed with salt water, then dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC. The compound 4-benzyloxy-N-[1,3-dimethyl-2-oxo-6-(3-propoxyphenoxy)benzimidazol-5-yl]benzenesulfonamide (2.30 g, 4.01 mmol, 44%) was obtained as yellow solid. LC/MS (ESI) m/z: 574.2 [M+1] ⁺ .

[00813] Step 4: Preparation of 4-(benzyloxy)-N-(1,3-dimethyl-2-oxo-6-(3-propoxyphenoxy)-2,3-dihydro-1H-benzo[d]imidazole-5- yl)-N-((2-(trimethylsilyl)ethoxy)methyl)benzenesulfonamide

[00814] To a solution of 4-benzyloxy-N-[1,3-dimethyl-2-oxo-6-(3-propoxyphenoxy)benzimidazol-5-yl]benzenesulfonamide (2.30 g, 4.01 mmol, 1.00 eq.) in tetrahydrofuran (30 ml) was added sodium hydride (208 mg, 5.21 mmol, 60% purity, 1.30 eq.) at 0°C for 30 minutes, then trimethylsiliethoxymethyl chloride (870 mg, 5 .21 mmol, 1.30 eq.). The reaction mixture was stirred at 25°C for 12 hours. Water (50 ml) was added to the mixture, then extracted with ethyl acetate (50 ml×3), the organic layers were washed with water, then dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate 10/1-1/1). The compound obtained was 4-benzyloxy-N-[1,3-dimethyl-2-oxo-6-(3-propoxyphenoxy)benzimidazol-5-yl]-N-(2-trimethylsilylethoxymethyl)benzenesulfonamide (2.70 g, 3.84 mmol, yield 96%) as a yellow solid. LC/MS (ESI) m/z: 704.2 [M+1] ⁺ .

[00815] Step 5: Preparation of N-(1,3-dimethyl-2-oxo-6-(3-propoxyphenoxy)-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-4-hydroxy -N-((2-(trimethylsilyl)ethoxy)methyl)benzenesulfonamide

[00816] Mixture of 4-benzyloxy-N-[1,3-dimethyl-2-oxo-6-(3-propoxyphenoxy)benzimidazol-5-yl]-N-(2-trimethylsilylethoxymethyl)benzenesulfonamide (2.70 g, 3 .84 mmol, 1.00 eq.), palladium on activated carbon (3.84 mmol, 10% purity, 1.00 eq.) in methanol (30 ml) was degassed and purged with nitrogen gas 3 times, and then the mixture was stirred at 25°C in an atmosphere of hydrogen gas for 3 hours. The mixture was filtered through a celite pad and the filtrate was concentrated to give the product. The compound N-[1,3-dimethyl-2-oxo-6-(3-propoxyphenoxy)benzimidazol-5-yl]-4-hydroxy-N-(2-trimethylsilylethoxymethyl)benzenesulfonamide (2.30 g, 3.75) was obtained. mmol, yield 98%) as a white solid. LC/MS (ESI) m/z: 636.3 [M+23] ⁺ .

[00817] Example of the synthesis of tert-butyl (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-(((2S,4R)- 4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy) ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin- 1-carboxylate and tert-butyl (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-(((2S,4R)-4 -hydroxy-1-((S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy )ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-1 -carboxylate

[00818] Step 1: Preparation of tert-butyl (2S,4R)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-hydroxypyrrolidine-1-carboxylate.

[00819] To a solution of tert-butyl (2S,4R)-4-hydroxy-2-(hydroxymethyl)pyrrolidine-1-carboxylate (25 g, 115.07 mmol, 1 eq.) in dichloromethane (400 mL) was added triethylamine ( 23.29 g, 230.14 mmol, 32.03 mL, 2 eq.) and N,N-dimethylpyridin-4-amine (1.41 g, 11.51 mmol, 0.1 eq.) and then added tert-butyldimethylsilyl chloride (18.21 g, 120.82 mmol, 1.05 eq.) at 0°C, the mixture was stirred at 25°C for 48 hours. The solution was evaporated in a water bath under reduced pressure using a rotary evaporator. This crude product was purified by silica gel column chromatography (EA:PE = 0:1 to 1:5) to give the compound tert-butyl (2S,4R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]- 4-hydroxypyrrolidine-1-carboxylate (40 g) as a colorless oil. ¹ H-NMR (400 MHz, CD ₃ OD) δ 4.45 (s, 1H), 4.00-3.91 (m, 1H), 3.56-3.53 (m, 2H), 3, 40 (s, 2H), 2.23-2.15 (m, 2H), 2.00-1.90 (m, 1H), 1.45 (s, 9H), 0.89 (s, 9H) , 0.01 (s, 6H).

[00820] Step 2: Preparation of tert-butyl (2S,4R)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(2-(2-((tetrahydro-2H-pyran-2-yl) oxy)ethoxy)ethoxy)pyrrolidine-1-carboxylate

[00821] To a solution of tert-butyl (2S,4R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-hydroxypyrrolidine-1-carboxylate (40 g, 120.66 mmol, 1 eq.) in tetrahydrofuran (500 ml) was added sodium hydride (7.24 g, 180.98 mmol, 60% in mineral oil, 1.5 eq.) at 0°C. The reaction mixture was stirred at 25°C for 1 hour. 2-(2-tetrahydropyran-2-yloxyethoxy)ethyl-4-methylbenzenesulfonate (45.71 g, 132.72 mmol, 1.1 eq.) was then added and the reaction mixture was stirred at 25°C for an additional 12 hours. The reaction mixture was quenched with saturated aqueous ammonium chloride (200 ml), then extracted with ethyl acetate (100 ml x 3). The organic layers were combined and evaporated in vacuo to leave a residue. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 1/0 to 2/1) to give the product. This gave tert-butyl (2S,4R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]pyrrolidine-1-carboxylate (40.8 g, 83.79 mmol, 68% yield) as a light yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.65-4.64 (m, 1H), 3.90-3.85 (m, 4H), 3.70-3.39 (m, 12H) , 2.23-2.13 (m, 1H), 2.10-1.92 (m, 1H), 1.91-1.49 (m, 6H), 1.45 (s, 9H), 0 .88 (s, 9H), 0.03 (s, 6H).

[00822] Step 3: Preparation of tert-butyl (2S,4R)-2-(hydroxymethyl)-4-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)pyrrolidine- 1-carboxylate

[00823] To a solution of tert-butyl (2S,4R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]pyrrolidine-1-carboxylate (10.8 g, 21.44 mmol, 1 eq.) in tetrahydrofuran (125 ml) was added tetrabutylammonium fluoride (1 M, 23.6 ml, 1.1 eq.) at 25°C. The mixture was stirred at 25°C for 12 hours. The solvent was removed in vacuo to leave a residue. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 10/1 to 1/1). The product was obtained tert-butyl (2S,4R)-2-(hydroxymethyl)-4-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]pyrrolidine-1-carboxylate (6.35 g, 16.30 mmol, yield 76%) as a light yellow oil.

[00824] Step 4: Preparation of ((2S,4R)-1-methyl-4-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)pyrrolidin-2-yl) methanol

[00825] To a solution of tert-butyl (2S,4R)-2-(hydroxymethyl)-4-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]pyrrolidine-1-carboxylate (20 g, 51.35 mmol, 1 eq.) in tetrahydrofuran (350 ml) was added lithium aluminum hydride (4.87 g, 128.38 mmol, 2.5 eq.) at 25°C. The mixture was stirred at 60°C for 16 hours. The reaction mixture was quenched with water (10 ml) before adding celite (20 g). The mixture was filtered and the filtrate was collected, then evaporated in vacuo to give [(2S,4R)-1-methyl-4-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]pyrrolidin-2-yl]methanol (14 d) in the form of a light yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.64-4.63 (m, 1H), 4.01-3.97 (m, 1H), 3.87-3.85 (m, 2H) , 3.70-3.46 (m, 9H), 3.40-3.38 (m, 2H), 2.63-2.61 (m, 1H), 2.40-2.36 (m, 1H), 2.33 (s, 3H), 2.13-2.02 (m, 1H), 1.88-1.43 (m, 8H).

[00826] Step 5: Preparation of tert-butyl-4-((S-4-((benzyloxy)carbonyl)-3-(cyanomethyl)piperazin-1-yl)-2-(((2S,4R)-1- methyl 4-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)pyrrolidin-2-yl)methoxy)-5,8-dihydropyrido[3,4-d]pyrimidine -7(6H)-carboxylate

[00827] To a solution of [(2S,4R)-1-methyl-4-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]pyrrolidin-2-yl]methanol (6.22 g, 20.49 mmol, 1.2 equiv.) and tert-butyl-4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-2-chloro-6,8-dihydro-5H-pyrido[3, 4-d]pyrimidine-7-carboxylate (9 g, 17.08 mmol, 1 eq.) in dioxane (180 ml) was added methanesulfonato(2-dicyclohexylphosphino-2',6'-di-and-propoxy-1,1 '-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (1.43 g, 1.71 mmol, 0.1 eq.) and cesium carbonate (16.69 g , 51.23 mmol, 3 eq.) under nitrogen atmosphere. The reaction mixture was stirred at 90°C for 6 hours under a nitrogen atmosphere. The solvent was removed in vacuo to obtain the crude product. The crude product was purified by silica gel column chromatography (ethyl acetate) to give the product. The product tert-butyl-4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-2-[[(2S,4R)-1-methyl-4-[2-(2 -tetrahydropyran-2-yloxyethoxy)ethoxy]pyrrolidin-2-yl]methoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-7-carboxylate (7.4 g, 6.63 mmol, yield 39%, purity 71%) as a brown solid. LC/MS (ESI) m/z: 794.5 [M+1] ⁺ .

[00828] Step 6: Preparation of benzyl(S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-hydroxyethoxy)ethoxy)-1-methylpyrrolidine-2) -yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazine-1-carboxylate

[00829] To a solution of tert-butyl-4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-2-[[(2S,4R)-1-methyl-4-[2 -(2-tetrahydropyran-2-yloxyethoxy)ethoxy]pyrrolidin-2-yl]methoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-7-carboxylate (7 g, 8.82 mmol , 1 eq.) in dichloromethane (100 ml) was added trifluoroacetic acid (25.13 g, 220.42 mmol, 16.3 ml, 25 eq.). The mixture was stirred at 25°C for 2 hours. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (100 ml), then extracted with dichloromethane (30 ml×3). The combined organic layers were evaporated in vacuo to remove solvent to obtain the crude product. The crude product was purified by preparative HPLC. The product was obtained benzyl(2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-(2-hydroxyethoxy)ethoxy]-1-methylpyrrolidin-2-yl]methoxy]- 5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (2.76 g, 4.13 mmol, 47% yield, 91% purity) as light yellow solid. LC/MS (ESI) m/z: 610.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.40-7.37 (m, 5H), 5.19 (s, 2H), 4.65 (s, 1H), 4.37 (dd, J =11.2 Hz, J=4.4 Hz, 1H), 4.18-2.58 (m, 26H), 2.46 (s, 3H), 2.38 (dd, J=9.6 Hz , J=6.0 Hz, 1H), 2.01-1.95 (m, 3H).

[00830] Step 7: Preparation of benzyl(S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-hydroxyethoxy)ethoxy)-1-methylpyrrolidine-2- yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-1-carboxylate

[00831] To a solution of benzyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-(2-hydroxyethoxy)ethoxy]-1-methylpyrrolidin-2-yl] methoxy]-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (2.2 g, 3.61 mmol, 1 eq.) and 1-bromonaphthalene ( 1.34 g, 6.49 mmol, 0.9 ml, 1.8 eq.) in dioxane (50 ml) added methanesulfonato(2-dicyclohexylphosphino-2',6'-di-isopropoxy-1,1'-biphenyl )(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (301.8 mg, 0.36 mmol, 0.1 eq.) and cesium carbonate (3.53 g, 10. 82 mmol, 3 eq.) under nitrogen atmosphere. The mixture was stirred at 90°C for 12 hours under a nitrogen atmosphere. The reaction mixture was quenched with water (50 ml) and extracted with ethyl acetate (40 ml×3). The organic layers were combined and evaporated in vacuo to leave a residue. The residue was purified by silica gel column chromatography (dichloromethane/methanol 10/1) to give the product. The product obtained was benzyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-(2-hydroxyethoxy)ethoxy]-1-methylpyrrolidin-2-yl]methoxy]- 7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (1.15 g, 1.39 mmol, 38% yield, purity 89%) as a yellow oil. LC/MS (ESI) m/z: 736.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.22-8.20 (m, 1H), 7.87-7.85 (m, 1H), 7.62 (d, J=8.0 Hz , 1H), 7.51-7.36 (m, 8H), 7.15 (d, J=6.8 Hz, 1H), 5.21 (s, 2H), 4.70 (s, 1H) , 4.38-4.10 (m, 8H), 3.76-3.31 (m, 14H), 3.10-2.72 (m, 6H), 2.47 (s, 3H), 2 .39-2.35 (m, 1H), 2.14-2.08 (m, 1H), 2.03-1.96 (m, 1H).

[00832] Step 8: Preparation of 2-((S)-4-(2-(((2S,4R)-4-(2-(2-hydroxyethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy) -7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[00833] To a solution of benzyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-(2-hydroxyethoxy)ethoxy]-1-methylpyrrolidin-2-yl] methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (1.38 g, 1.88 mmol, 1 eq.) and ammonium hydroxide (1.82 g, 12.98 mmol, 2 ml, 25% purity, 6.92 eq.) in methanol (60 ml) and tetrahydrofuran (3 ml) added palladium on activated carbon catalyst, (200 mg, purity 10%). The mixture was degassed and charged with hydrogen, then stirred at 25°C under a hydrogen atmosphere (15 psi) for 12 hours. Tetrahydrofuran (40 ml) was added to the reaction mixture and filtered. The organic solvent was removed in vacuo to give 2-[(2S)-4-[2-[[(2S,4R)-4-[2-(2-hydroxyethoxy)ethoxy]-1-methylpyrrolidin-2-yl]methoxy] -7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-2-yl]acetonitrile (1.1 g) as a light yellow solid substances. LC/MS (ESI) m/z: 602.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.23-8.21 (m, 1H), 7.87-7.85 (m, 1H), 7.62 (d, J=8.4 Hz , 1H), 7.51-7.49 (m, 2H), 7.43 (t, J=8.4 Hz, 1H), 7.15-7.13 (m, 1H), 4.41- 4.39 (m, 1H), 4.26 (s, 2H), 4.22 (dd, J=11.2 Hz, J=6.4 Hz, 1H), 4.03 (d, J=12 .0 Hz, 1H), 3.88 (d, J=12.0 Hz, 1H), 3.76-3.72 (m, 3H), 3.67-3.54 (m, 6H), 3 .45-2.85 (m, 12H), 2.57-2.55 (m, 2H), 2.47 (s, 3H), 2.39 (dd, J=9.6 Hz, J=6 .0 Hz, 1H), 2.11-1.84 (m, 2H).

[00834] Step 9: Preparation of tert-butyl(S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-hydroxyethoxy)ethoxy)-1-methylpyrrolidine) -2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-1-carboxylate

[00835] To a solution of 2-[(2S)-4-[2-[[(2S,4R)-4-[2-(2-hydroxyethoxy)ethoxy]-1-methylpyrrolidin-2-yl]methoxy]-7 -(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-2-yl]acetonitrile (800 mg, 1.33 mmol, 1 eq.) in tetrahydrofuran (30 ml) and water (30 ml) were added sodium bicarbonate (223.4 mg, 2.66 mmol, 0.1 ml, 2 eq.) and di-tert-butyl dicarbonate (1.45 g, 6.65 mmol , 1.5 ml, 5 eq.). The mixture was stirred at 25°C for 6 hours. The reaction mixture was quenched by adding water (20 ml) and extracted with ethyl acetate (30 ml×3). The organic phase was washed with brine (30 ml x 2), evaporated in vacuo to obtain the crude product. The crude product was purified by silica gel column chromatography (dichloromethane/methanol 10/1). The product was obtained: tert-butyl(2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-(2-hydroxyethoxy)ethoxy]-1-methylpyrrolidin-2-yl]methoxy ]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (670 mg, 0.95 mmol, 72% yield) as a light yellow solid. LC/MS (ESI) m/z: 702.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.23-8.20 (m, 1H), 7.87-7.85 (m, 1H), 7.62 (d, J=8.4 Hz , 1H), 7.51-7.49 (m, 2H), 7.43 (t, J=8.4 Hz, 1H), 7.15-7.14 (m, 1H), 4.63 ( s, 1H), 4.41-4.39 (m, 1H), 4.26-4.22 (m, 3H), 4.12 (d, J=12.0 Hz, 1H), 3.75 -3.31 (m, 14H), 3.10-2.76 (m, 6H), 2.48 (s, 3H), 2.40 (dd, J=9.6 Hz, J=6.0 Hz, 1H), 2.37-2.05 (m, 2H), 1.52 (s, 9H).

[00836] Step 10: Preparation of tert-butyl(S)-2-(cyanomethyl)-4-(2-(((2S,4R)-1-methyl-4-(2-(2-(tosyloxy)ethoxy) ethoxy)pyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-1-carboxylate

[00837] To a solution of tert-butyl(2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-(2-hydroxyethoxy)ethoxy]-1-methyl-pyrrolidine- 2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (300 mg, 0.43 mmol , 1 eq.) in dichloromethane (5 ml) was added triethylamine (129.8 mg, 1.28 mmol, 0.2 ml, 3 eq.), p-toluenesulfonyl chloride (163 mg, 0.85 mmol, 2 eq.) and dimethylaminopyridine (10.4 mg, 0.09 mmol, 0.2 eq.). The mixture was stirred at 25°C for 12 hours. The solvent was removed in vacuo to leave a residue. The residue was purified by preparative TLC (silica, dichloromethane/methanol = 10/1). The product was obtained: tert-butyl(2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-1-methyl-4-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]pyrrolidine -2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (233 mg, 0.27 mmol, yield 64%) as a light yellow solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.22-8.20 (m, 1H), 7.87-7.85 (m, 1H), 7.81-7.75 (m, 2H) , 7.62 (d, J=8.0 Hz, 1H), 7.51-7.49 (m, 2H), 7.43 (t, J=8.0 Hz, 1H), 7.35 ( d, J=8.0 Hz, 2H), 7.15-7.12 (m, 1H), 4.63 (s, 1H), 4.43-4.39 (m, 1H), 4.27 -3.93 (m, 8H), 3.72-3.27 (m, 12H), 3.07-2.75 (m, 6H), 2.49 (s, 3H), 2.45 (s , 3H), 2.41-2.36 (m, 1H), 2.09-1.98 (m, 2H), 1.52 (s, 9H).

[00838] Step 11: Preparation of tert-butyl (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-(((2S,4R )-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidin-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl) phenoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl) piperazine-1-carboxylate and tert-butyl (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-(((2S,4R) -4-hydroxy-1-((S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy )ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazine -1-carboxylate

[00839] To a solution of tert-butyl(2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-1-methyl-4-[2-[2-(p-tolyl-sulfonyloxy) ethoxy]ethoxy]pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (250 mg, 0.29 mmol, 1 eq.) and (2S,4R)-4-hydroxy-N-[[2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]methyl]-1-[3 -methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (152.9 mg, 0.31 mmol, 1.05 eq.) in acetonitrile (8 ml) was added cesium carbonate (190, 3 mg, 0.58 mmol, 2 eq.). The mixture was stirred at 80°C for 14 hours. The mixture was extracted with ethyl acetate (30 ml×3) after adding water (30 ml). The combined organic phases were evaporated in vacuo to leave a residue. The residue was purified by preparative TLC (silica, dichloromethane/methanol = 10/1). The product containing two isomers (255 mg) was obtained as a yellow solid. The product was then further purified using SFC. The product tert-butyl(2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-[[[(2S,4R)-4- hydroxy-1-[(2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidin-2-carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy] ethoxy]ethoxy]-1-methylpyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-1- carboxylate (80 mg, 0.06 mmol, 21% yield, 92% purity) as a yellow oil. The product tert-butyl(2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-[[[(2S,4R)-4- hydroxy-1-[(2S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidin-2-carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy] ethoxy]ethoxy]-1-methylpyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-1- carboxylate (120 mg, 0.096 mmol, 33% yield, 95% purity) as a light yellow solid. LC/MS (ESI) m/z: 1182.7 [M+1] ⁺ .

[00840] Example of the synthesis of tert-butyl(S)-4-(4-((benzyloxy)carbonyl)-3-(cyanomethyl)piperazin-1-yl)-2-chloro-5,8-dihydropyrido[3,4- d]pyrimidine-7(6H)-carboxylate

[00841] Step 1: Preparation of 1-benzyl-4-(tert-butyl)(R)-2-(hydroxymethyl)piperazine-1,4-dicarboxylate

[00842] To a solution of tert-butyl(3R)-3-(hydroxymethyl)piperazine-1-carboxylate (5.00 g, 23.12 mmol, 1.00 eq.) in ethyl acetate (50 ml) and water (50 ml ) sodium bicarbonate (5.83 g, 69.00 mmol, 3.00 eq.) was added in one portion, then benzyl carbonochloridate (5.94 g, 35.00 mmol, 1.51 eq.) was slowly added to the solution while stirring at 0°C for 30 minutes. The resulting solution was stirred at a temperature of 10°C for 5 hours. The organic layer was separated from the reaction solution and washed with water (10 ml). The aqueous phase was extracted with ethyl acetate (100 ml). The organic layer was collected, combined, washed with water (30 ml x 3), brine (30 ml), dried over sodium sulfate, concentrated under reduced pressure to obtain a yellow liquid. The yellow liquid was purified by silica gel column chromatography (petroleum ether/ethyl acetate 3/1 to 1/1) to give O1-benzyl-O4-tert-butyl(2R)-2-(hydroxymethyl)piperazine-1,4 -dicarboxylate (8.00 g, 22.83 mmol, 99% yield) as a colorless liquid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.33-7.24 (m, 5H), 5.08 (s, 2H), 4.18 (br s, 1H), 3.87 (br s , 2H), 3.57 (br s, 2H), 3.12-2.78 (m, 2H), 1.97 (s, 2H), 1.60-1.57 (m, 1H), 1 .40 (s, 9H).

[00843] Step 2: Preparation of 1-benzyl-4-(tert-butyl)(R)-2-(((methylsulfonyl)oxy)methyl)piperazine-1,4-dicarboxylate

[00844] To a solution of O1-benzyl-O4-tert-butyl (2R)-2-(hydroxymethyl)piperazine-1,4-dicarboxylate (6.31 g, 18.01 mmol, 1.00 eq.) in dichloromethane ( 100 mL) triethylamine (5.47 g, 54.00 mmol, 3.00 eq.) was added in one portion. Methyl sulfonyl chloride (3.09 g, 27.00 mmol, 1.50 eq.) was then added to the solution. The resulting solution was stirred at 25°C for 2 hours. The solution was concentrated under reduced pressure to obtain O1-benzyl O4-tert-butyl (2R)-2-(methylsulfonyloxymethyl)piperazine-1,4-dicarboxylate (6.66 g) as a yellow liquid.

[00845] Step 3: Preparation of 1-benzyl-4-(tert-butyl)(S)-2-(cyanomethyl)piperazine-1,4-dicarboxylate

[00846] To a solution of O1-benzyl-O4-tert-butyl (2R)-2-(methylsulfonyloxymethyl)piperazine-1,4-dicarboxylate (2.45 g, 5.72 mmol, 1.00 eq.) in N, To N-dimethylacetamide (10 mL), cyanosodium (560 mg, 11.00 mmol, 2.00 eq.) was added in one portion. The solution was stirred at 55°C for 24 hours. The solution was poured into ethyl acetate (200 ml), the solution was washed with water (50 ml×2). The organic layer was separated and collected, washed with brine (50 ml), concentrated under reduced pressure to obtain a yellow liquid. The yellow liquid was purified by column chromatography (silica, petroleum ether/ethyl acetate = 10/1 to 4/1) to give O1-benzyl-O4-tert-butyl(2S)-2-(cyanomethyl)piperazine-1,4 -dicarboxylate (1.00 g, 2.00 mmol, 40% yield) as a colorless liquid. LC/MS (ESI) m/z: 304.1 [M-55] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.39 (s, 5H), 5.19 (s, 2H), 4.58 (br s, 1H), 4.15-3.86 (m, 3H), 3.24-2.48 (m, 5H), 1.51 (s, 9H).

[00847] Step 4: Preparation of benzyl(S)-2-(cyanomethyl)piperazine-1-carboxylate

[00848] To a solution of O1-benzyl-O4-tert-butyl(2S)-2-(cyanomethyl)piperazine-1,4-dicarboxylate (900 mg, 2.50 mmol, 1.00 eq.) in dichloromethane (10 ml ) trifluoroacetic acid (3.08 g, 27.00 mmol, 10.79 eq.) was added slowly. The solution was stirred at 10°C for 2 hours. The solution was concentrated under reduced pressure to give benzyl(2S)-2-(cyanomethyl)piperazine-1-carboxylate (911 mg, 2.44 mmol, 97% yield, trifluoroacetic acid salt) as a yellow liquid.

[00849] Step 5: Preparation of tert-butyl(S)-4-(4-((benzyloxy)carbonyl)-3-(cyanomethyl)piperazin-1-yl)-2-chloro-5,8-dihydropyrido[3, 4-d]pyrimidine-7(6H)-carboxylate

[00850] To a solution of benzyl(2S)-2-(cyanomethyl)piperazine-1-carboxylate (646 mg, 2.49 mmol, 1.00 eq., trifluoroacetic acid salt) and diisopropylethylamine (1.29 g, 9.96 mmol, 4.00 eq.) in dimethyl sulfoxide (20 ml) was added tert-butyl2,4-dichloro-5,6-dihydropyrido[3,4-d]pyrimidine-7(8H)-carboxylate (758 mg, 2.49 mmol, 1.00 eq.) in one serving. The resulting solution was stirred at 50°C for 9 hours. The reaction solution was diluted with ethyl acetate (200 ml) and water (100 ml). The organic layer was separated, collected and washed with water (50 ml×2) and brine (50 ml), dried over anhydrous sodium sulfate, concentrated under reduced pressure to obtain a yellow liquid. The yellow liquid was purified by column chromatography (silica, petroleum ether/ethyl acetate = 10/1 to 1/1) to give tert-butyl-4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazine-1- yl]-2-chloro-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-7-carboxylate (1.10 g, 2.09 mmol, 84% yield) as a yellow liquid. LC/MS (ESI) m/z: 527.1 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.46-7.32 (m, 5H), 5.26-5.14 (m, 2H), 4.67 (d, J=17.6 Hz , 2H), 4.51-4.42 (m, 1H), 4.21-4.05 (m, 2H), 3.93-3.75 (m, 2H), 3.40 (d, J =10.8 Hz, 2H), 3.12 (dt, J=3.2, 12.4 Hz, 1H), 2.97-2.51 (m, 3H), 1.61 (s, 2H) , 1.50 (s, 9H).

[00851] Example of the synthesis of tert-butyl(S)-2-(cyanomethyl)-4-(2-(((2S,4R)-1-methyl-4-(2-(2-(2-(tosyloxy)ethoxy )ethoxy)ethoxy)pyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-1- carboxylate

[00852] Step 1: Preparation of tert-butyl (2S,4R)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(2-(2-(2-((tetrahydro-2H-pyran-2 -yl)oxy)ethoxy)ethoxy)ethoxy)pyrrolidine-1-carboxylate

[00853] To a mixture of tert-butyl(2S,4R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-hydroxypyrrolidine-1-carboxylate (15 g, 45.25 mmol, 1.1 eq. ) in tetrahydrofuran (30 ml), add sodium hydride (3.29 g, 82.37 mmol, 60% purity, 2 eq.) at 25°C, stir for 1 hour, then add 2-[2-(2- tetrahydropyran-2-yloxyethoxy)ethoxy]ethyl-4-methylbenzenesulfonate (15.98 g, 41.13 mmol, 1 eq.), the mixture was stirred at 25°C for 12 hours. The residue was poured into ice water (100 ml) and stirred for 0.5 minutes. The aqueous phase was extracted with ethyl acetate (70 ml×3). The combined organic phase was washed with brine (50 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate 5:1 to 2:1). The compound tert-butyl(2S,4R)-2-[[tert-butyl (dimethyl)silyl]oxymethyl]-4-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]pyrrolidine-1 was obtained -carboxylate (18.2 g, 33.22 mmol, 81% yield) as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.63 (t, J=3.4 Hz, 1H), 4.51-4.14 (m, 1H), 4.03-3.78 (m , 4H), 3.69-3.66 (m, 6H), 3.65-3.55 (m, 6H), 3.54-3.47 (m, 2H), 3.44-3.38 (m, 1H), 2.27-2.13 (m, 1H), 2.00-1.78 (m, 2H), 1.77-1.65 (m, 2H), 1.60 (br d, J=8.2 Hz, 2H), 1.45 (br s, 9H), 1.22 -1.17 (m, 1H), 0.87 (s, 9H), 0.02 (br s , 6H).

[00854] Step 2: Preparation of tert-butyl(2S,4R)-2-(hydroxymethyl)-4-(2-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy )ethoxy)pyrrolidine-1-carboxylate

[00855] To a mixture of tert-butyl (2S,4R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy] pyrrolidine-1-carboxylate (18.2 g, 33.22 mmol, 1 eq.) in tetrahydrofuran (80 ml) was added tetrabutylammonium (1 M, 33.22 ml, 1 eq.) at 20°C. The reaction mixture was stirred at 20°C for 2 hours. The reaction mixture was concentrated in vacuo to obtain a residue. The residue was purified by silica gel column chromatography (5:1 to 1:1 petroleum ether:ethyl acetate) to give tert-butyl(2S,4R)-2-(hydroxymethyl)-4-[2-[2-(2 -tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]pyrrolidine-1-carboxylate (9 g, 20.76 mmol, 62% yield) as a yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.66-4.62 (m, 1H), 4.39 (br s, 1H), 4.03 (br s, 1H), 3.92-3.82 (m, 2H), 3.75-3.70 ( m, 1H), 3.70-3.68 (m, 2H), 3.67 (d, J=2.0 Hz, 4H), 3.64-3.61 (m, 2H), 3.60-3.54 (m, 4H), 3.54-3.44 (m, 2H), 3.44-3.37 (m, 1H), 2.20-2.10 (m, 1H), 1.89-1.79 (m, 1H), 1.78-1.66 (m, 2H), 1.64-1.57 (m, 2H), 1.53 ( br d, J=6.6 Hz, 2H), 1.47 (s, 1H).

[00856] Step 3: Preparation of ((2S,4R)-1-methyl-4-(2-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)ethoxy)pyrrolidine -2-yl)methanol

[00857] To a mixture of tert-butyl(2S,4R)-2-(hydroxymethyl)-4-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]pyrrolidine-1-carboxylate (9 g, 20.76 mmol, 1 eq.) in tetrahydrofuran (150 ml), lithium aluminum hydride (3.94 g, 103.80 mmol, 5 eq.) was added, then the reaction mixture was stirred at 70°C for 12 hours. The mixture was cooled to 25°C. Tetrahydrofuran (200 ml) was added, water (5 ml) was added slowly, then the mixture was filtered and concentrated in vacuo. The crude product was purified by chromatography on silica gel (petroleum ether:ethyl acetate 3:1 to 0:1), then (tetrahydrofuran:methanol = 10:1) 0.1% ammonium hydroxide to give [(2S,4R)-1 -methyl-4-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]pyrrolidin-2-yl]methanol (5.2 g, 14.97 mmol, 72% yield) as a colorless oil . ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.67-4.60 (m, 1H), 4.42-4.34 (m, 1H), 4.04-3.96 (m, 1H) , 3.91-3.74 (m, 2H), 3.72-3.64 (m, 7H), 3.64-3.54 (m, 4H), 3.54-3.47 (m, 1H), 3.43-3.36 (m, 2H), 2.77-2.61 (m, 1H), 2.42-2.36 (m, 2H), 2.35-2.32 ( m, 3H), 2.29-2.05 (m, 2H), 1.94-1.67 (m, 4H), 1.65-1.55 (m, 2H).

[00858] Step 4: Preparation of tert-butyl-4-((S-4-((benzyloxy)carbonyl)-3-(cyanomethyl)piperazin-1-yl)-2-(((2S,4R)-1- methyl 4-(2-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)ethoxy)pyrrolidin-2-yl)methoxy)-5,8-dihydropyrido[3, 4-d]pyrimidine-7(6H)-carboxylate

[00859] To a solution of tert-butyl-4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-2-chloro-6,8-dihydro-5H-pyrido[3,4- d]pyrimidine-7-carboxylate (1.7 g, 3.23 mmol, 1 eq.) and [(2S,4R)-1-methyl-4-[2-[2-(2-tetrahydropyran-2-yloxyethoxy )ethoxy]ethoxy]pyrrolidin-2-yl]methanol (1.40 g, 4.03 mmol, 1.25 eq.) in dioxane (20 ml) was added methanesulfonato(2-dicyclohexylphosphino-2,6-di-isopropoxy- 1,1-biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(ii) (270 mg, 0.32 mmol, 0.1 eq.) and cesium carbonate (3.15 g, 9 .68 mmol, 3 eq.), the mixture was stirred at 90°C for 2 hours under a nitrogen atmosphere. Water (100 ml) was added. The aqueous phase was extracted with ethyl acetate (150 ml×3). The combined organic phase was washed with brine (50 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate 4:1 to 0:1) and then purified by silica gel chromatography (dichloromethane:methanol = 10:1) to give the product. The compound tert-butyl-4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-2-[[(2S,4R)-1-methyl-4-[2-[2 -(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]pyrrolidin-2-yl]methoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-7-carboxylate (2.3 g) as brown butter. LC/MS (ESI) m/z: 838.6 [M+1] ⁺ .

[00860] Step 5: Preparation of benzyl(S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)-1 -methylpyrrolidin-2)-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazine-1-carboxylate

[00861] To a mixture of tert-butyl-4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-2-[[(2S,4R)-1-methyl-4-[2 -[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]pyrrolidin-2-yl]methoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-7-carboxylate (2, 5 g, 2.98 mmol, 1 eq.) in dichloromethane (20 ml), trifluoroacetic acid (3.08 g, 27.01 mmol, 2 ml, 9.05 eq.) was added, then the reaction mixture was stirred at 20°C within 5 hours. The residue was poured into a saturated potassium carbonate solution and stirred for 0.5 minutes. Lithium hydrate was then added to adjust the pH to 12 with stirring for 20 minutes, then the aqueous phase was extracted with dichloromethane and methanol (10:1, 50 ml×3), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by semipreparative reverse phase HPLC to give the compound benzyl(2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-(2-hydroxyethoxy)ethoxy] ethoxy]-1-methylpyrrolidin-2-yl]methoxy]-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (1.3 g, 1.99 mmol, yield 67%) as a brown oil. LC/MS (ESI) m/z: 676.3 [M+23] ⁺ .

[00862] Step 6: Preparation of benzyl(S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)-1 -methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-1-carboxylate

[00863] To a solution of benzyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-1-methylpyrrolidine -2-yl]methoxy]-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (1 g, 1.53 mmol, 1 eq.) and 1 -bromonaphthalene (633 mg, 3.06 mmol, 0.4 ml, 2 eq.) in dioxane (10 ml) was added methanesulfonato(2-dicyclohexylphosphino-2,6-di-isopropoxy-1,1-biphenyl)(2- amino-1,1-biphenyl-2-yl)palladium(ii) (128 mg, 0.15 mmol, 0.1 eq.) and cesium carbonate (1.50 g, 4.59 mmol, 3 eq.). The mixture was stirred at 90°C for 8 hours under a nitrogen atmosphere. Water (10 ml) was added. The aqueous phase was extracted with ethyl acetate (15 ml×3). The combined organic phase was washed with brine (10 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (dichloromethane:methanol = 50:1 to 10:1) to give benzyl(2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4- [2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-1-methylpyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d ]pyrimidin-4-yl]piperazine-1-carboxylate (310 mg, 0.40 mmol, 26% yield) as a brown solid. LC/MS (ESI) m/z: 780.5 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.24-8.15 (m, 1H), 7.90-7.82 (m, 1H), 7.61 (d, J=8.1 Hz , 1H), 7.54-7.46 (m, 2H), 7.46-7.33 (m, 6H), 7.14 (d, J=7.4 Hz, 1H), 5.21 ( s, 2H), 4.70 (br s, 1H), 4.39 (br s, 1H), 4.34-4.24 (m, 2H), 4.23-4.15 (m, 2H) , 4.06 (br s, 1H), 3.95 (br d, J=11.5 Hz, 1H), 3.76-3.72 (m, 2H), 3.70-3.54 (m , 10H), 3.51-3.41 (m, 2H), 3.32 (br d, J=11.9 Hz, 3H), 3.10-3.01 (m, 1H), 3.00 -2.79 (m, 4H), 2.79-2.72 (m, 1H), 2.48 (br s, 3H), 2.39 (br s, 1H), 2.09 (br dd, J=4.3, 7.5 Hz, 1H), 2.02-1.97 (m, 1H), 1.86 (td, J=3.3, 6.6 Hz, 2H), 0.92 -0.75 (m, 1H).

[00864] Step 7: Preparation of 2-((S)-4-(2-(((2S,4R)-4-(2-(2(2-hydroxyethoxy)ethoxy)ethoxy)-1-methylpyrrolidine-2- yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[00865] To a solution of benzyl(2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-1-methylpyrrolidine -2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (310 mg, 0.40 mmol, 1 eq.) in methanol (10 ml), palladium on carbon (30 mg, 10% purity) and ammonium hydroxide (273 mg, 2.34 mmol, 0.3 ml, 30% purity, 5.88 eq.) were added. ) in a nitrogen atmosphere. The suspension was degassed in vacuum and purged with hydrogen several times. The mixture was stirred under a hydrogen atmosphere (15 psi) at 20°C for 5 hours. The mixture was concentrated in vacuo. The compound 2-[(2S)-4-[2-[[(2S,4R)-4-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-1-methylpyrrolidin-2-yl]methoxy] was obtained. -7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-2-yl]acetonitrile (180 mg, 0.26 mmol, 67% yield , purity 95%) as a yellow solid. LC/MS (ESI) m/z: 646.4 [M+1] ⁺ .

[00866] Step 8: Preparation of tert-butyl(S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy) -1-methylpyrrolidin)-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazine-1-carboxylate

[00867] To a solution of 2-[(2S)-4-[2-[[(2S,4R)-4-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-1-methylpyrrolidin-2-yl ]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-2-yl]acetonitrile (150 mg, 0.23 mmol, 1 eq.) in dichloromethane (3 ml) was added di-tert-butyl dicarbonate (507 mg, 2.32 mmol, 0.5 ml, 10 eq.) and triethylamine (71 mg, 0.70 mmol, 0.1 ml, 3 eq.), the reaction mixture was stirred at 25°C for 12 hours. The mixture was filtered and concentrated in vacuo. The reaction mixture was purified by preparative TLC (dichloromethane:methanol = 10:1). The compound tert-butyl(2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-1-methylpyrrolidin- 2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (130 mg, 0.16 mmol , yield 71%, purity 95%) as a yellow oil. LC/MS (ESI) m/z: 746.3 [M+1] ⁺ .

[00868] Step 9: Preparation of tert-butyl(S)-2-(cyanomethyl)-4-(2-(((2S,4R)-1-methyl-4-(2-(2-(2-(tosyloxy) )ethoxy)ethoxy)ethoxy)pyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin- 1-carboxylate

[00869] To a solution of tert-butyl(2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-1 -methylpyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-1-carboxylate (130 mg, 0 .17 mmol, 1 eq.) in dichloromethane (2 ml), triethylamine (53 mg, 0.52 mmol, 3 eq.) and paratoluenesulfonyl chloride (66 mg, 0.35 mmol, 2 eq.) and dimethylaminopyridine (4 mg, 0.034 mmol, 0.2 eq.). The reaction mixture was stirred at 25°C for 12 hours. The mixture was filtered and concentrated in vacuo. The reaction mixture was purified by preparative TLC (dichloromethane:methanol = 10:1). The compound tert-butyl(2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-1-methyl-4-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy] was obtained ethoxy]ethoxy]pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (100 mg 0.11 mmol, yield 61%, purity 95%) as a yellow oil. LC/MS (ESI) m/z: 900.4 [M+1] ⁺ .

[00870] Synthesis example of (2S,4R)-1-((S)-18-(tert-butyl)-16-oxo-5,8,11,14-tetraoxa-2,17-diazanonadecan-19-oil) -4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[00871] Step 1: Preparation of tert-butyl ((S)-16-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl) phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-17,17-dimethyl-14-oxo-3,6,9,12-tetraoxa-15-azaoctadecyl)(methyl)carbamate

[00872] A mixture of 2-[2-[2-[2-[2-[tert-butoxycarbonyl(methyl)amino]ethoxy]ethoxy]ethoxy]ethoxy]acetic acid (450 mg, 1.23 mmol, 1.00 eq .), (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]pyrrolidine-2-carboxamide (592 mg, 1.23 mmol, 1.00 eq., hydrochloride), hydroxybenzotriazole (199 mg, 1.48 mmol, 1.20 eq.), 1-(3 -Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (283 mg, 1.48 mmol, 1.20 eq.) in dichloromethane (10 ml) and diisopropylethylamine (318 mg, 2.46 mmol, 430 μl, 2.00 eq.) was degassed and purged with nitrogen 3 times, and then the mixture was stirred at 20°C for 12 hours under a nitrogen atmosphere. The reaction mixture was diluted with water (30 ml), adjusted to pH=2 with hydrochloric acid (1 M) and then extracted with ethyl acetate (30 ml x 2). The combined organic layers were washed with saturated sodium bicarbonate solution (50 ml×1), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (100-90% ethyl acetate in methanol) to give tert-butylN-[2-[2-[2-[2-[2-[[(1S)-1-[(2S, 4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl] amino]-2-oxoethoxy]ethoxy]ethoxy]ethoxy]ethyl]-N-methylcarbamate (650 mg, 0.82 mmol, 67% yield) as a colorless oil. LC/MS (ESI) m/z: 792.5 [M+1] ⁺ ; ¹ H-NMR (400 MG c ,, CDCl ₃ ) δ 8.68 (s, 1H), 7.53-7.29 (m, 6H), 5.17-4.99 (m, 1H), 4 .77 (t, J=8.0 Hz, 1H), 4.58-4.47 (m, 2H), 4.25-3.96 (m, 3H), 3.79-3.50 (m , 16H), 3.44-3.32 (m, 2H), 2.91 (s, 3H), 2.65-2.50 (m, 4H), 2.14-2.01 (m, 1H ), 1.52-1.42 (m, 12H), 1.08 (s, 9H).

[00873] Step 2: Preparation of (2S,4R)-1-((S)-18-(tert-butyl)-16-oxo-5,8,11,14-tetraoxa-2,17-diazanonadecane-19- oil)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[00874] To a solution of tert-butyl N-[2-[2-[2-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S )-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl]amino]-2-oxoethoxy]ethoxy]ethoxy]ethoxy]ethyl] -N-methylcarbamate (650 mg, 0.82 mmol, 1 eq.) in dichloromethane (4 ml) was added hydrochloric acid in dioxane (4 M, 3 ml). The mixture was stirred at 20°C for 0.5 hour. The reaction mixture was concentrated under reduced pressure to obtain the compound (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-[2-[2-[2-(methylamino)ethoxy ]ethoxy]ethoxy]ethoxy]acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (600 mg, hydrochloride) as green oil. LC/MS (ESI) m/z: 692.2 [M+1] ⁺ .

[00875] Example of the synthesis of tert-butyl(R)-(2-(2-hydroxyethoxy)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamate

[00876] Step 1: Preparation of tert-butyl R)-(1-(4-bromophenyl)-2-hydroxyethyl)carbamate

[00877] To a solution of (2R)-2-amino-2-(4-bromophenyl)ethanol (5 g, 19.80 mmol, 1 eq., hydrochloride) and triethylamine (6.01 g, 59.40 mmol, 8 .27 mL, 3 eq.) in dichloromethane (80 mL), di-tert-butyl dicarbonate (4.66 g, 21.35 mmol, 4.91 mL, 1.08 eq.) was added. The reaction mixture was stirred at 20°C for 16 hours. The reaction solution was washed with water (100 ml), dried over sodium sulfate and then concentrated in vacuo to give tert-butyl-N-[(1R)-1-(4-bromophenyl)-2-hydroxyethyl]carbamate (6.07 g, 18.85 mmol, yield 95%, purity 98%) as a white solid. LC/MS (ESI) m/z: 338.1 [M+23] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.50 (d, J=8.4 Hz, 2H), 7.24 (d, J=8.8 Hz, 2H), 4.79 (t, J=5.6 Hz, 1H), 4.48 (br s, 1H), 3.47 (br dd, J=6.0, 10.4 Hz, 2H), 1.36 (s, 9H).

[00878] Step 2: Preparation of ethyl (R)-2-(2-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)ethoxy)acetate

[00879] To a solution of tert-butyl-N-[(1R)-1-(4-bromophenyl)-2-hydroxyethyl]carbamate (4 g, 12.65 mmol, 1 equiv.) and diacetoxyrhodium (279 mg, 1. 27 mmol, 0.1 eq.) in dichloromethane (150 ml), ethyl 2-diazoacetate (5.41 g, 37.95 mmol, 4.97 ml, 3 eq.) was added dropwise at 0°C. The reaction mixture was stirred at 20°C for 16 hours. The reaction mixture was concentrated in vacuo to obtain a residue. The residue was purified by silica gel column chromatography (0-10% ethyl acetate in petroleum ether) to give ethyl 2-[(2R)-2-(4-bromophenyl)-2-(tert-butoxycarbonylamino)ethoxy]acetate (5.1 g ) as a white solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.48-7.43 (m, 2H), 7.24 (d, J=8.4 Hz, 2H), 5.79 (br s, 1H) , 4.87-4.62 (m, 1H), 4.25-4.24 (m, 2H), 4.06 (d, J=1.2 Hz, 2H), 3.80-3.73 (m, 1H), 3.69 (br d, J=4.8 Hz, 1H), 1.50-1.36 (m, 9H), 1.30-1.26 (m, 3H).

[00880] Step 3: Preparation of ethyl(R)-2-(2-((tert-butoxycarbonyl)amino)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane) -2-yl)phenyl)ethoxy)acetate

[00881] To a solution of ethyl 2-[(2R)-2-(4-bromophenyl)-2-(tert-butoxycarbonylamino)ethoxy]acetate (3 g, 7.46 mmol, 1 eq.) and 4,4,4 ',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (2.27 g, 8.95 mmol, 1.2 eq.) in dioxane (45 ml) was added dioxane (45 ml), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(ii) (436 mg, 0.60 mmol, 0.08 eq.) and potassium acetate (1.46 g, 14.92 mmol, 2 eq.). The reaction mixture was degassed and purged with nitrogen 3 times and then heated to 90°C for 16 hours. The reaction mixture was concentrated in vacuo to obtain a residue. The residue was purified by silica gel column chromatography (0-30% ethyl acetate in petroleum ether) to give ethyl 2-[(2R)-2-(tert-butoxycarbonylamino)-2-[4-(4,4,5,5-tetramethyl -1,3,2-dioxaborolan-2-yl)phenyl]ethoxy]acetate (3.35 g) as a yellow oil.

[00882] Step 4: Preparation of ethyl (R)-2-(2-((tert-butoxycarbonyl)amino)-2-(4-(4-methylthiazol-5-yl)phenyl)ethoxy)acetate

[00883] To a solution of ethyl 2-[(2R)-2-(tert-butoxycarbonylamino)-2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl]ethoxy]acetate (3.35 g, 7.46 mmol, 1 eq.) and 5-bromo-4-methylthiazole (2.65 g, 14.91 mmol, 2 eq.) in dioxane (60 ml) were added [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(ii) (545 mg, 0.75 mmol, 0.1 eq.), potassium carbonate (2.06 g, 14.91 mmol, 2 eq.) and water (12 ml). The reaction mixture was degassed and purged with nitrogen 3 times, and then the mixture was stirred at 85°C for 6 hours. The reaction mixture was concentrated in vacuo to remove most of the solvents. Water (40 ml) was added and the mixture was extracted with ethyl acetate (40 ml×2). The organic layer was dried over sodium sulfate and then concentrated in vacuo to obtain a residue. The residue was purified by silica gel flash chromatography (0-100% ethyl acetate in petroleum ether) to give ethyl 2-[(2R)-2-(tert-butoxycarbonylamino)-2-[4-(4-methylthiazol-5-yl )phenyl]ethoxy]acetate (1.6 g, 3.80 mmol, 51% yield) as a light yellow gum. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.70 (s, 1H), 7.42 (s, 4H), 5.86 (br s, 1H), 4.86 (br s, 1H), 4.25 (q, J=7.2 Hz, 2H), 4.10 (s, 2H), 3.88-3.71 (m, 2H), 2.55 (s, 3H), 1.45 (br s, 9H), 1.31 (t, J=7.2 Hz, 3H).

[00884] Step 5: Preparation of tert-butyl(R)-(2-(2-hydroxyethoxy)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamate

[00885] To a suspension of lithium aluminum hydride (131 mg, 3.45 mmol, 1 eq.) in tetrahydrofuran (20 ml) was added a solution of ethyl 2-[(2R)-2-(tert-butoxycarbonylamino)-2-[4-( 4-methylthiazol-5-yl)phenyl]ethoxy]acetate (1.45 g, 3.45 mmol, 1 eq.) in tetrahydrofuran (20 ml) at minus 5°C. The reaction mixture was stirred at 0°C for 1 hour. Lithium aluminum hydride (131 mg, 3.45 mmol, 1 eq.) was then added and the reaction mixture was stirred at 0° C. for an additional 1 hour. Water (20 ml) was added and the mixture was extracted with ethyl acetate (30 ml). The organic layer was dried over sodium sulfate and then concentrated in vacuo to obtain a residue. The residue was purified by silica gel flash chromatography (0-200% ethyl acetate in petroleum ether) to give tert-butyl-N-[(1R)-2-(2-hydroxyethoxy)-1-[4-(4-methylthiazol-5 -yl)phenyl]ethyl]carbamate (780 mg, 2.06 mmol, 59% yield) as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.68 (s, 1H), 7.48-7.35 (m, 4H), 5.33 (br d, J=7.6 Hz, 1H) , 4.90 (br s, 1H), 3.81-3.76 (m, 1H), 3.76-3.68 (m, 3H), 3.67-3.61 (m, 1H), 3.60-3.53 (m, 1H), 2.54 (s, 3H), 2.16-1.93 (m, 1H), 1.44 (br s, 9H).

[00886] Example of synthesis of (2S,4R)-4-hydroxy-1-(3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxylic acid

[00887] Step 1: Preparation of 2-(3-methylisoxazol-5-yl)acetic acid

[00888] To a solution of 3,5-dimethylisoxazole (15 g, 154.46 mmol, 15 mL, 1 eq.) in tetrahydrofuran (150 mL), n-butyllithium (2.5 M, 77 mL, 1.25 eq.) at minus 78°C under nitrogen atmosphere, the mixture was stirred at minus 55°C for 30 minutes, and then the mixture was bubbled with carbon dioxide for 30 minutes, the mixture was stirred at 25°C for 1 hour. The mixture was quenched with saturated ammonium chloride solution (50 ml), and the mixture was extracted with ethyl acetate (50 ml). The aqueous phase was adjusted to pH=2 with aqueous hydrochloric acid (2 M), the mixture was extracted with ethyl acetate (50 ml, three times), the organic phase was dried with anhydrous sodium sulfate, filtered and the filtrate was concentrated to give 2-(3-methylisoxazole-5 -yl)acetic acid (10 g, 70.86 mmol, 46% yield) as a brown solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 12.74 (br s, 1H), 6.24 (s, 1H), 3.83 (s, 2H), 2.20 (s, 3H) .

[00889] Step 2: Preparation of methyl 2-(3-methylisoxazol-5-yl) acetate

[00890] Thionyl chloride (12.65 g, 106.29 mmol, 7.71 ml, 1.5 eq.) at 0°C, and the mixture was stirred at 50°C for 4 hours. The mixture was concentrated to give the crude product. The said crude product was diluted with ethyl acetate (200 ml) and washed with water (200 ml) and then with saturated aqueous sodium bicarbonate (50 ml) and then with brine (50 ml), the organic phase was dried with anhydrous (compound), filtered, and the filtrate condensed to give methyl 2-(3-methylisoxazol-5-yl) acetate (10 g, 64.45 mmol, 91% yield) as a brown oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 6.11 (s, 1H), 3.80 (s, 2H), 3.76 (s, 3H), 2.30 (s, 3H).

[00891] Step 3: Preparation of methyl 3-methyl-2-(3-methylisoxazol-5-yl)butanoate

[00892] To a solution of methyl 2-(3-methylisoxazol-5-yl)acetate (10 g, 64.45 mmol, 1 eq.) in tetrahydrofuran (100 ml) was added sodium hydride (3.87 g, 96.68 mmol , 60% purity, 1.5 eq.) at 0°C, and then 2-iodopropane (13.15 g, 77.34 mmol, 7.74 mL, 1.2 eq.) was added at 0°C, mixture stirred at 25°C for 2 hours. Additional 2-iodopropane (2.55 g, 15.00 mmol, 1.5 ml) was added and the mixture was stirred at 25°C for 10 hours. The mixture was quenched with aqueous hydrochloric acid (1 M, 300 ml) and the mixture was extracted with ethyl acetate (200 ml, three times), the organic phase was dried with anhydrous sodium sulfate, filtered and the filtrate was concentrated to give methyl-3-methyl-2-(3-methylisoxazole -5-yl)butanoate (13 g) as a brown oil.

[00893] Step 4: Preparation of 3-methyl-2-(3-methylisoxazol-5-yl)butanoic acid

[00894] To a solution of methyl 3-methyl-2-(3-methylisoxazol-5-yl)butanoate (12.7 g, 64.39 mmol, 1 eq.) in methanol (90 ml) and water (60 ml) sodium hydroxide was added. (12.88 g, 321.96 mmol, 5 eq.), the mixture was stirred at 25°C for 2 hours. The mixture was concentrated to remove methanol and then the residue was diluted with water (200 ml) and extracted with ethyl acetate (200 ml), the aqueous phase was adjusted to pH=3 with aqueous hydrochloric acid (2 M) and then the mixture was extracted with dichloromethane (200 ml, three times ). The organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the crude product as a brown oil. This material was purified by preparative flash HPLC, the acetonitrile fraction was removed and the residue was extracted with dichloromethane (300 ml×5). The organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the product 3-methyl-2-(3-methylisoxazol-5-yl)butanoic acid (7.5 g, 40.94 mmol, 63% yield) as a white solid substances. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 6.26 (s, 1H), 3.58 (d, J=8.7 Hz, 1H), 2.33-2.23 (m, 1H ), 2.21 (s, 3H), 0.95 (d, J=6.7 Hz, 3H), 0.82 (d, J=6.8 Hz, 3H).

[00895] Step 5: Preparation of methyl (2S,4R)-4-hydroxy-1-(3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxylate

[00896] To a solution of methyl (2S,4R)-4-hydroxypyrrolidine-2-carboxylate (1.19 g, 6.55 mmol, 1 eq., hydrochloride), 3-methyl-2-(3-methylisoxazole-5- yl)butanoic acid (1.2 g, 6.55 mmol, 1 eq.) and triethylamine (1.99 g, 19.65 mmol, 2.74 ml, 3 eq.) in N,N-dimethylformamide (20 ml ) o-(7-azabenzotriazol-1-yl)-n,n,n',n'-tetramethyluronium hexafluorophosphate (2.74 g, 7.21 mmol, 1.1 eq.) was added. The reaction mixture was stirred at 25°C for 40 minutes. The reaction mixture was dissolved in ethyl acetate (50 ml) and then washed with brine (50 ml). The organic layer was dried over sodium sulfate and then concentrated in vacuo to give a residue (3 g). The residue was purified by preparative HPLC to give methyl(2S,4R)-4-hydroxy-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxylate (1.64 g, 5 .28 mmol, yield 80%) in the form of a light brown gum. LC/MS (ESI) m/z: 311.1 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 6.20-6.04 (m, 1H), 4.68-4.50 (m, 2H), 3.89 (dd, J=4, 3, 10.8 Hz, 1H), 3.78-3.69 (m, 3H), 3.69-3.57 (m, 2H), 2.47-2.34 (m, 1H), 2 .28 (d, J=9.1 Hz, 4H), 2.12-2.09 (m, 1H), 1.12-0.95 (m, 3H), 0.89 (d, J=6 ,8 Hz, 3H).

[00897] Step 6: Preparation of (2S,4R)-4-hydroxy-1-(3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxylic acid

[00898] To a solution of methyl(2S,4R)-4-hydroxy-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxylate (1.2 g, 3.87 mmol, 1 eq.) in tetrahydrofuran (12 ml), water (12 ml) and lithium hydroxide monohydrate (487 mg, 11.60 mmol, 3 eq.) were added. The reaction mixture was stirred at 25°C for 2 hours. The pH of the mixture was adjusted to 2 with 1 N hydrochloric acid. The mixture was extracted with ethyl acetate (40 ml×2). The organic layer was dried over sodium sulfate and then concentrated in vacuo to give (2S,4R)-4-hydroxy-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxylic acid (1.02 g, 3.44 mmol, yield 89.02%) as a colorless gum. LC/MS (ESI) m/z: 297.1 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 6.23-6.14 (m, 1H), 4.57-4.39 (m, 2H), 3.90-3.67 (m, 2H ), 3.66-3.57 (m, 1H), 2.50-2.32 (m, 1H), 2.31-2.21 (m, 4H), 2.13-2.02 (m , 1H), 1.12-0.96 (m, 3H), 0.91-0.78 (m, 3H).

[00899] Example of the synthesis of tert-butyl((R)-1-((2S,4R)-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl) pyrrolidin-2-yl)-3-(4-(4-methylthiazol-5-yl)phenyl)-1-oxo-5,8,11-trioxa-2-azatridecan-13-yl)(methyl)carbamate and tert -butyl((3R)-1-((2S,4R)-4-hydroxy-1-(3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidin-2-yl)-3-( 4-(4-methylthiazol-5-yl)phenyl)-1-oxo-5,8,11-trioxa-2-azatridecan-13-yl)(methyl)carbamate

[00900] Step 1: Preparation of 2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl 4-methylbenzenesulfonate.

[00901] 3,4-Dihydro-2H-pyran (496 mg, 5.76 mmol, 1.5 eq.) and p-toluenesulfonic acid (33 mg, 0.19 mmol, 0.05 eq.). The reaction solution was stirred at 20°C for 2 hours. Triethylamine (0.1 ml) was added and the reaction mixture was concentrated in vacuo to leave a residue. The residue was purified by flash chromatography on silica gel (0-25% ethyl acetate in petroleum ether) to give the product. 2-(2-tetrahydropyran-2-yloxyethoxy)ethyl-4-methylbenzenesulfonate (1.15 g, 3.34 mmol, 86.91% yield) was obtained as a colorless oil. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.81 (d, J=8.3 Hz, 2H), 7.35 (d, J=8.1 Hz, 2H), 4.62- 4.58 (m, 1H), 4.20-4.16 (m, 2H), 3.90-3.77 (m, 2H), 3.74-3.69 (m, 2H), 3. 64-3.59 (m, 2H), 3.57-3.46 (m, 2H), 2.46 (s, 3H), 1.88-1.77 (m, 1H), 1.75- 1.67 (m, 1H), 1.57-1.48 (m, 4H).

[00902] Step 2: Preparation of tert-butyl((1R)-1-(4-(4-methylthiazol-5-yl)phenyl)-2-(2-(2-(2-((tetrahydro-2H-pyran -2-yl)oxy)ethoxy)ethoxy)ethoxy)ethyl)carbamate

[00903] To a solution of tert-butyl-N-[(1R)-2-(2-hydroxyethoxy)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamate (430 mg, 1.14 mmol, 1 eq.) in tetrahydrofuran (4 ml), sodium hydride (91 mg, 2.28 mmol, 60% purity, 2 eq.) was added at 0°C. The reaction mixture was stirred at 20°C for 0.5 hour. A solution of 2-(2-tetrahydropyran-2-yloxyethoxy)ethyl-4-methylbenzenesulfonate (393 mg, 1.14 mmol, 1 eq) in tetrahydrofuran (6 ml) was then added and the reaction mixture was stirred at 55°C for 12 hours. The mixture was quenched with water (30 ml) and extracted with ethyl acetate (30 ml, three times), the organic phase was dried with anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the crude product. This crude product was purified by preparative TLC (dichloromethane:methanol = 10:1) to give the product, tert-butyl-N-[(1R)-1-[4-(4-methylthiazol-5-yl)phenyl]-2- [2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]ethyl]carbamate (85 mg, 0.12 mmol, 10.8% yield, 79.4% purity) as a brown oil.

[00904] Step 3: Preparation of (R)-2-(2-(2-(2-amino-2-(4-(4-methylthiazol-5-yl)phenyl)ethoxy)ethoxy)ethoxy)ethane-1- ola

[00905] To a solution of tert-butyl-N-[(1R)-1-[4-(4-methylthiazol-5-yl)phenyl]-2-[2-[2-(2-tetrahydropyran-2-yloxyethoxy) ethoxy]ethoxy]ethyl]carbamate (84 mg, 0.15 mmol, 1 eq.) in dichloromethane (2 ml) added hydrochloric acid/methanol (4 M, 0.5 ml, 13.11 eq.) at 20°C . The reaction mixture was stirred at 20°C for 3 hours. The reaction mixture was concentrated in vacuo. The compound 2-[2-[2-[(2R)-2-amino-2-[4-(4-methylthiazol-5-yl)phenyl]ethoxy]ethoxy]ethoxy]ethanol (61 mg, 0.15 mmol) was obtained , 99.3% yield, hydrochloride) as a yellow oil. LC/MS (ESI) m/z: 367.2 [M+1] ⁺ .

[00906] Step 4: Preparation of (2S,4R)-4-hydroxy-N-((R)-2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)-1-(4-(4- methylthiazol-5-yl)phenyl)ethyl)-1-(3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide

[00907] To a solution of 2-[2-[2-[(2R)-2-amino-2-[4-(4-methylthiazol-5-yl)phenyl]ethoxy]ethoxy]ethoxy]ethanol (98 mg, 0 .24 mmol, 1 equiv., hydrochloride), (2S,4R)-4-hydroxy-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxylic acid (76 mg , 0.26 mmol, 1.05 eq.), 1-hydroxybenzotriazole (43 mg, 0.32 mmol, 1.3 eq.) and N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (61 mg, 0. 32 mmol, 1.3 eq.) in N,N-dimethylformamide (2 ml) was added diisopropylethylamine (94 mg, 0.73 mmol, 3 eq.). The reaction mixture was stirred at 20°C for 15 hours. The mixture was diluted with water (30 ml) and extracted with dichloromethane (30 ml, twice), the organic phase was dried with anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the crude product. This crude product was purified by preparative TLC (dichloromethane:methanol = 10:1) to obtain the product. We obtained (2S,4R)-4-hydroxy-N-[(1R)-2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-1-[4-(4-methylthiazol-5-yl) phenyl]ethyl]-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide, 180 mg total, as a pale yellow oil. LC/MS (ESI) m/z: 645.1 [M+1] ⁺ .

[00908] Step 5: Preparation of (3R)-1-((2S,4R)-4-hydroxy-1-(3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidin-2-yl) -3-(4-(4-methylthiazol-5-yl)phenyl)-1-oxo-5,8,11-trioxa-2-azatridecan-13-yl 4-methylbenzenesulfonate

[00909] To a solution of (2S,4R)-4-hydroxy-N-[(1R)-2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-1-[4-(4-methylthiazol- 5-yl)phenyl]ethyl]-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (100 mg, 0.11 mmol, 1 eq.) in dichloromethane ( 3 ml) triethylamine (22 mg, 0.21 mmol, 2 eq.), dimethylaminopyridine (1.31 mg, 10.70 µmol, 0.1 eq.) was added, followed by 4-methylbenzene-1-sulfonyl chloride (41 mg , 0.21 mmol, 2 eq.), the mixture was stirred at 20°C for 10 hours. The mixture was concentrated to give the crude product. This crude product was purified by preparative TLC (dichloromethane:methanol = 10:1) to obtain the product. The compound 2-[2-[2-[(2R)-2-[[(2S,4R)-4-hydroxy-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine was obtained -2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethoxy]ethoxy]ethoxy]ethyl 4-methylbenzenesulfonate (40 mg, 0.04 mmol, 40.2% yield, purity 85.9%) as brown oil. LC/MS (ESI) m/z: 799.0 [M+1] ⁺ .

[00910] Step 6: Preparation of (2S,4R)-4-hydroxy-1-(3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)-N-((R)-13-(4- (4-methylthiazol-5-yl)phenyl)-5,8,11-trioxa-2-azatridecan-13-yl)pyrrolidin-2-carboxamide

[00911] 2-[2-[2-[(2R)-2-[[(2S,4R)-4-hydroxy-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl solution ]pyrrolidine-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethoxy]ethoxy]ethoxy]ethyl 4-methylbenzenesulfonate (100 mg, 0.13 mmol, 1 eq.) in methylamine/ethanol (4 ml, purity 30%), the mixture was stirred at 50°C for 3 hours. The mixture was concentrated to give the product (2S,4R)-4-hydroxy-N-[(1R)-2-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]-1-[4-(4 -methylthiazol-5-yl)phenyl]ethyl]-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (90 mg) as a pale yellow oil. LC/MS (ESI) m/z: 658.2 [M+1] ⁺ .

[00912] Step 7: Preparation of tert-butyl((3R)-1-((2S,4R)-4-hydroxy-1-(3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidin- 2-yl)-3-(4-(4-methylthiazol-5-yl)phenyl)-1-oxo-5,8,11-trioxa-2-azatridecan-13-yl)(methyl)carbamate

[00913] To a solution of (2S,4R)-4-hydroxy-N-[(1R)-2-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]-1-[4-(4 -methylthiazol-5-yl)phenyl]ethyl]-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (82 mg, 0.12 mmol, 1 eq.) in dichloromethane (3 ml), triethylamine (38 mg, 0.37 mmol, 3 eq.) and di-tert-butyl dicarbonate (54 mg, 0.25 mmol, 57 μl, 2 eq.) were added, the mixture was stirred at 20°C within 5 hours. The mixture was concentrated to give the crude product. This crude product was purified by preparative TLC (dichloromethane:methanol = 10:1) to give tert-butyl N-[2-[2-[2-[(2R)-2-[[(2S,4R)-4-hydroxy -1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethoxy]ethoxy] ethoxy]ethyl]-N-methylcarbamate (75 mg, 98.95 µmol, 79.4% yield) as a pale yellow oil. LC/MS (ESI) m/z: 780.1 [M+23] ⁺ .

[00914] Step 8: Preparation of tert-butyl((R)-1-((2S,4R)-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl) butanoyl)pyrrolidin-2-yl)-3-(4-(4-methylthiazol-5-yl)phenyl)-1-oxo-5,8,11-trioxa-2-azatridecan-13-yl)(methyl)carbamate and tert-butyl((3R)-1-((2S,4R)-4-hydroxy-1-(3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidin-2-yl)-3 -(4-(4-methylthiazol-5-yl)phenyl)-1-oxo-5,8,11-trioxa-2-azatridecan-13-yl)(methyl)carbamate

[00915] Tert-butyl N-[2-[2-[2-[(2R)-2-[[(2S,4R)-4-hydroxy-1-[3-methyl-2-(3-methylisoxazol- 5-yl)butanoyl]pyrrolidin-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethoxy]ethoxy]ethoxy]ethyl]-N-methylcarbamate (75 mg, 0.10 mmol, 1 eq.) was purified using SFC. The compound tert-butyl N-[2-[2-[2-[(2R)-2-[[(2S,4R)-4-hydroxy-1-[(2R)-3-methyl-2-(3) was obtained -methylisoxazol-5-yl)butanoyl]pyrrolidin-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethoxy]ethoxy]ethoxy]ethyl]-N-methylcarbamate (30 mg, 35.50 µmol, yield 35.9%, purity 89.7%) and tert-butyl N-[2-[2-[2-[(2R)-2-[[(2S,4R)-4-hydroxy -1-[(2S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidin-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl] ethoxy]ethoxy]ethoxy]ethyl]-N-methylcarbamate (28 mg, 34.36 µmol, 34.7% yield, 93% purity) as colorless oils.

[00916] Example of the synthesis of 3-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N -(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethyl)-N-methylpropanamide (Typical connection 13)

[00917] Step 1: Preparation of tert-butyl (2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy )ethyl)(methyl)carbamate

[00918] To a stirred solution of 2,2,5-trimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl4-methylbenzenesulfonate (370 mg, 0.73 mmol) 2-(2.6 -dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (200 mg, 0.73 mmol) and potassium carbonate (200 mg, 1.5 mmol) in N,N-dimethylformamide (10 ml). The resulting mixture was stirred at 60°C overnight. The reaction mixture was partitioned between water (20 ml) and ethyl acetate (15 ml). The organic layer was collected, washed with water (20 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by preparative TLC (eluting with 8% methanol in dichloromethane) to give tert-butyl (2-(2- (2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethyl)(methyl) carbamate (180 mg, 47%). LC/MS (ESI) m/z: 520.5 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.45 (s, 9H), 2.13-2.17 (m, 1H), 2.73-2.93 (m, 6H), 3.40 (s, 2H), 3.60-3.65 (m, 4H), 3.71-3.73 (m, 2H), 3.91 (t, J=4.4 Hz, 2H), 4, 25 (t, J=3.6 Hz, 3H), 4.96 (dd, J=5.2 Hz, J=12.0 Hz, 1H), 7.23 (dd, J=2.0 Hz, J=8.0 Hz, 1H), 7.36 (d, J=2.0 Hz, 1H), 7.78 (d, J=8.4 Hz, 1H), 8.09 (s, 1H) .

[00919] Step 2: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(2-(2-(2-(methylamino)ethoxy)ethoxy)ethoxy)isoindoline-1,3-dione hydrochloride

[00920] Tert-Butyl(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethyl) solution (methyl)carbamate (90 mg, 0.17 mmol) in 4M hydrogen chloride in dioxane (3 ml) was stirred at room temperature for 1 hour. Volatiles were removed under reduced pressure to give 2-(2,6-dioxopiperidin-3-yl)-5-(2-(2-(2-(methylamino)ethoxy)ethoxy)ethoxy)isoindoline-1,3-dione hydrochloride in the form of HCl salt.

[00921] Step 3: Preparation of 3-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino) -N-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethyl)-N-methylpropanamide

[00922] To a stirred solution of 2-(2,6-dioxopiperidin-3-yl)-5-(2-(2-(2-(methylamino)ethoxy)ethoxy)ethoxy)isoindoline-1,3-dione hydrochloride (0 ,17 mmol), 3-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)propanoic acid (50 mg, 0.09 mmol) in N,N-dimethylformamide (5 ml) was added N-ethyl-N-isopropylpropan-2-amine (36 mg, 0.28 mmol) and (2-(7-aza-1H -benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) (71 mg, 0.19 mmol) at 0°C. The resulting mixture was stirred at room temperature for 24 hours. The reaction mixture was partitioned between ethyl acetate (20 ml) and water (20 ml). The organic layer was collected, washed with water (20 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by preparative TLC to give 3-((4-(4-acetylpiperazin-1-yl)-6 -chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-(2-(2-(2-((2-(2,6-dioxopiperidin-3 -yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethyl)-N-methylpropanamide (50 mg, 57%) as a white solid. LC/MS (ESI) m/z: 941.0 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 2.01-2.07 (m, 4H), 2.54-2.75 (m, 4H), 2.87-2.99 (m, 4H), 3.47-3.77 (m, 16H), 4.18-4.28 (m, 6H), 5.09-5.14 (m, 1H), 7.07-7.08 ( m, 1H), 7.22-7.27 (m, 2H), 7.32-7.36 (m, 2H), 7.43-7.48 (m, 2H), 7.80-7, 84 (m, 2H), 8.04-8.09 (m, 2H), 11.12 (s, 1H).

[00923] Example of the synthesis of 3-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N -(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethyl)-N-methylpropanamide (Typical connection 111)

[00924] Step 1: Preparation of tert-butyl (2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy )ethyl)(methyl)carbamate

[00925] To a solution of 2-(2,6-dioxo-3-piperidyl)-4-fluoroisoindoline-1,3-dione (151 mg, 0.55 mmol, 1.2 eq.) and tert-butyl N-[ 2-[2-(2-aminoethoxy)ethoxy]ethyl]-N-methylcarbamate (120 mg, 0.46 mmol, 1 eq.) in dimethyl sulfoxide (5 ml) was added N,N-diisopropylethylamine (77 mg, 0.60 mmol, 0.1 ml, 1.3 eq.). The mixture was stirred at 80°C for 12 hours. The mixture was diluted with water (20 ml) and the aqueous phase was extracted with ethyl acetate (20 ml×3). The combined organic phase was washed with brine (20 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative TLC (petroleum ether/ethyl acetate 1/1) to give tert-butyl-N-[2-[2-[2-[[2-(2,6-dioxo-3-piperidyl)-1 ,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethyl]-N-methylcarbamate (100 mg, 0.19 mmol, 40% yield, 96% purity) as a yellow oil. LC/MS (ESI) m/z: 541.1 [M+23] ⁺ .

[00926] Step 2: Preparation of 2-(2,6-dioxopiperidin-3-yl)-4-((2-(2-(2-(methylamino)ethoxy)ethoxy)ethyl)amino)isoindoline-1,3- Diona

[00927] A dioxane hydrochloride solution (4 M, 10 mL, 207.42 eq.) was added to tert-butyl-N-[2-[2-[2-[[2-(2,6-dioxo-3- piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethyl]-N-methylcarbamate (100 mg, 0.19 mmol, 1 eq.), and the mixture was stirred at 20°C for 12 hours. The mixture was concentrated in vacuo to give 2-(2,6-dioxo-3-piperidyl)-4-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethylamino]isoindoline-1,3-dione (110 mg , hydrochloride) as a colorless oil.

[00928] Step 3: Preparation of tert-butyl-4-((7S)-6-chloro-2-((3-((2-(2-(2-((2-(2,6-dioxopiperidine-3 -yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl)(methyl)amino)-3-oxopropyl)amino)-8-fluoro-7-(2-fluoro-6-hydroxyphenyl) quinazolin-4-yl)piperazine-1-carboxylate

[00929] To a mixture of 2-(2,6-dioxo-3-piperidyl)-4-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethylamino]isoindoline-1,3-dione (120 mg, 0.22 mmol, 1 equiv., trifluoroacetate) and 3-[[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazoline -2-yl]amino]propanoic acid (127 mg, 0.22 mmol, 1 eq.) in N,N-dimethylformamide (5 ml) was added 1-hydroxybenzotriazole (60 mg, 0.45 mmol, 2 eq.), N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (86 mg, 0.45 mmol, 2 eq.) and N,N-diisopropylethylamine (145 mg, 1.13 mmol, 0.20 ml, 5 eq.). The mixture was stirred at 25°C for 12 hours. The mixture was diluted with water (20 ml), extracted with ethyl acetate (20 ml×3). The combined organic layer was washed with water (30 ml×2) and brine (30 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative thin layer chromatography (dichloromethane:methanol = 10:1) to give tert-butyl-4-[6-chloro-2-[[3-[2-[2-[2-[[2-(2, 6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethylmethylamino]-3-oxo-propyl]amino]-8-fluoro-7-(2-fluoro- 6-Hydroxyphenyl)quinazolin-4-yl]piperazine-1-carboxylate (150 mg, 0.15 mmol, 69% yield) as a light yellow solid. LC/MS (ESI) m/z: 964.4 [M+1] ⁺ .

[00930] Step 4: Preparation of 3-(((S)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-(piperazin-1-yl)quinazolin-2-yl) amino)-N-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl)-N -methylpropanamide

[00931] To a mixture of tert-butyl 4-[6-chloro-2-[[3-[2-[2-[2-[[2-(2,6-dioxo-3-piperidyl)-1,3- dioxoisoindolin-4-yl]amino]ethoxy]ethoxy]ethylmethylamino]-3-oxopropyl]amino]-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazolin-4-yl]piperazin-1-carboxylate (150 mg, 0.15 mmol, 1 eq.) in dichloromethane (4 ml) was added trifluoroacetic acid (1 ml). The mixture was stirred at 25°C for 1 hour. The mixture was concentrated under reduced pressure to give 3-[[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-piperazin-1-ylquinazolin-2-yl]amino]-N-[2 -[2-[2-[[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethyl]-N-methylpropanamide (150 mg , trifluoroacetate) as a green oil. LC/MS (ESI) m/z: 864.3 [M+1] ⁺ .

[00932] Step 5: Preparation of 3-(((S)-4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazoline-2- yl)amino)-N-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl) -N-methylpropanamide

[00933] To a mixture of 3-[[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-piperazin-1-ylquinazolin-2-yl]amino]-N-[2-[ 2-[2-[[2-(2,6-Dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethyl]-N-methylpropanamide (75 mg, 0 .08 mmol, 1 equiv., trifluoroacetate) and 2,6-lutidine (82 mg, 0.77 mmol, 0.09 ml, 10 equiv.) in dichloromethane (20 ml) was added prop-2-enoyl chloride (7 mg, 0.08 mmol, 0.006 ml, 1 eq.) in dichloromethane (5 ml) at minus 78°C. The mixture was stirred at minus 78°C for 30 minutes under a nitrogen atmosphere. The mixture was quenched with water (20 ml). The mixture was then extracted with dichloromethane (20 ml×2). The combined organic layer was washed with water (20 ml×2) and brine (20 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by semipreparative reverse phase HPLC. 3-[[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]amino]-N was obtained -[2-[2-[2-[[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethyl]-N-methylpropanamide (36.9 mg, 0.04 mmol, 51% yield, 99% purity) as a yellow solid. LC/MS (ESI) m/z: 918.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.09 (s, 1H), 10.47 (s, 1H), 8.10-8.00 (m, 1H), 7.94-7 .77 (m, 1H), 7.55-7.51 (m, 1H), 7.39-7.37 (m, 1H), 7.11-7.00 (m, 2H), 6.87 -6.74 (m, 3H), 6.59-6.56 (m, 1H), 6.18 (d, J=16.8 Hz, 1H), 5.75 (d, J=9.6 Hz, 1H), 5.04 (dd, J=4.8, 13.2 Hz, 1H), 4.25-4.13 (m, 3H), 3.85-3.68 (m, 17H) , 2.95-2.84 (m, 4H), 2.70-2.59 (m, 6H), 2.02-1.99 (m, 1H).

[00934] Example of the synthesis of 3-(((R)-4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazolin-2-yl) amino)-N-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl)-N -methylpropanamide (Typical compound 112)

[00935] Step 1: Preparation of tert-butyl-4-((7R)-6-chloro-2-((3-((2-(2-(2-((2-(2,6-dioxopiperidine-3 -yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl)(methyl)amino)-3-oxopropyl)amino)-8-fluoro-7-(2-fluoro-6-hydroxyphenyl) quinazolin-4-yl)piperazine-1-carboxylate

[00936] To a mixture of 2-(2,6-dioxo-3-piperidyl)-4-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethylamino]isoindoline-1,3-dione (120 mg, 0.22 mmol, 1 equiv., trifluoroacetate) and 3-[[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazoline -2-yl]amino]propanoic acid (127 mg, 0.22 mmol, 1 eq.) in N,N-dimethylformamide (5 ml) was added 1-hydroxybenzotriazole (60 mg, 0.45 mmol, 2 eq.), N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (86 mg, 0.45 mmol, 2 eq.) and N,N-diisopropylethylamine (145 mg, 1.13 mmol, 0.2 ml, 5 eq.). The mixture was stirred at 25°C for 4 hours. The mixture was diluted with water (20 ml). The mixture was then extracted with ethyl acetate (20 ml×3). The combined organic layer was washed with water (30 ml×2) and brine (30 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative thin layer chromatography (dichloromethane:methanol = 10:1). We obtained tert-butyl 4-[6-chloro-2-[[3-[2-[2-[2-[[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin- 4-yl]amino]ethoxy]ethoxy]ethylmethylamino]-3-oxopropyl]amino]-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazolin-4-yl]piperazine-1-carboxylate (160 mg, 0.16 mmol, yield 73%) as a light yellow oil. LC/MS (ESI) m/z: 964.4 [M+1] ⁺ .

[00937] Step 2: Preparation of 3-(((R)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-(piperazin-1-yl)quinazolin-2-yl) amino)-N-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl)-N -methylpropanamide

[00938] To a mixture of tert-butyl 4-[6-chloro-2-[[3-[2-[2-[2-[[2-(2,6-dioxo-3-piperidyl)-1,3- dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethylmethylamino]-3-oxopropyl]amino]-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazolin-4-yl]piperazine-1-carboxylate (160 mg, 0.16 mmol, 1 eq.) in dichloromethane (4 ml) was added trifluoroacetic acid (1 ml). The mixture was stirred at 25°C for 1 hour. The mixture was concentrated under reduced pressure to give 3-[[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-piperazin-1-ylquinazolin-2-yl]amino]-N-[2 -[2-[2-[[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethyl]-N-methylpropanamide (160 mg , trifluoroacetate) as a green oil. LC/MS (ESI) m/z: 864.3 [M+1] ⁺ .

[00939] Step 3: Preparation of 3-(((R)-4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazoline-2- yl)amino)-N-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethyl) -N-methylpropanamide

[00940] To a mixture of 3-[[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-piperazin-1-ylquinazolin-2-yl]amino]-N-[2-[ 2-[2-[[2-(2,6-Dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethyl]-N-methylpropanamide (80 mg, 0 .08 mmol, 1 equiv., trifluoroacetate) and 2,6-lutidine (87 mg, 0.81 mmol, 0.09 ml, 10 equiv.) in dichloromethane (20 ml) was added prop-2-enoyl chloride (7 mg, 0.08 mmol, 0.006 ml, 1 eq.) in dichloromethane (5 ml) at minus 78°C. The mixture was stirred at minus 78°C for 30 minutes under a nitrogen atmosphere. The mixture was quenched with water (20 ml). The mixture was then extracted with dichloromethane (20 ml×2). The combined organic layer was washed with water (20 ml×2) and brine (20 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by semipreparative reverse phase HPLC. 3-[[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]amino]-N was obtained -[2-[2-[2-[[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethyl]-N-methylpropanamide (45.3 mg, 0.04 mmol, 59% yield, 99% purity) as a yellow solid. LC/MS (ESI) m/z: 918.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.09 (s, 1H), 10.49 (s, 1H), 8.02-8.00 (m, 1H), 7.95-7 .80 (m, 1H), 7.55-7.51 (m, 1H), 7.40-7.38 (m, 1H), 7.12-7.01 (m, 2H), 6.88 -6.74 (m, 3H), 6.55-6.50 (m, 1H), 6.18 (d, J=16.8 Hz, 1H), 5.75 (d, J=9.6 Hz, 1H), 5.04 (dd, J=4.8, 13.6 Hz, 1H), 4.20-4.11 (m, 3H), 3.86-3.68 (m, 17H) , 2.95-2.84 (m, 4H), 2.71-2.55 (m, 6H), 2.02-2.00 (m, 1H).

[00941] Synthesis example of (E)-5-(2-(2-((5-(4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl) piperazin-1-yl)-5-oxopent-3-en-1-yl)oxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (typical compound 169 )

[00942] Step 1: Preparation of 2-(2-(3-(benzyloxy)propoxy)ethoxy)ethan-1-ol

[00943] To a solution of 2,2'-oxydiethanol (8 g, 74.9 mmol) in dry N,N-dimethylformamide (20 ml) was added sodium hydride (60% in mineral oil) (1.5 g, 37.4 mmol) at 0°C. The mixture was stirred at 50°C for 1 hour. 3-(Benzyloxy)propyl-4-methylbenzenesulfonate (4 g, 12.5 mmol) was then added at 50°C and the mixture was stirred at 70°C for 12 hours. The mixture was cooled to room temperature and partitioned between ethyl acetate (100 ml) and water (200 ml). The organic layer was collected, washed with brine (50 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by flash silica gel chromatography (eluting with 33-50% ethyl acetate in hexane to 2% methanol in dichloromethane ) to give 2-(2-(3-(benzyloxy)propoxy)ethoxy)ethanol (1.94 g, 61%) as a yellow oil.

[00944] Step 2: Preparation of 2-(2-(2-(3-(benzyloxy)propoxy)ethoxy)ethoxy)tetrahydro-2H-pyran

[00945] A mixture of 2-(2-(3-(benzyloxy)propoxy)ethoxy)ethanol (1.94 g, 7.63 mmol), 3,4-dihydro-2H-pyran (1.28 g, 15.25 mmol), pyridine-1-ium 4-methylbenzenesulfonate (960 mg, 3.81 mmol) and N,N-dimethylpyridin-4-amine (93 mg, 0.76 mmol) in dichloromethane (20 ml) were stirred under reflux refrigerator for 5 hours. The mixture was concentrated and the residue was partitioned between water (30 ml) and ethyl acetate (20 ml). The organic layer was collected, washed with brine (20 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by silica gel flash chromatography (eluting with 18-25% ethyl acetate in hexane) to give 2-( 2-(2-(3-(benzyloxy)propoxy)ethoxy)ethoxy)tetrahydro-2H-pyran (2.4 g, 88%) as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.48-1.64 (m, 4H), 1.68-1.75 (m, 1H), 1.79-1.85 (m, 1H) , 1.87-1.93 (m, 2H), 3.47-3.53 (m, 1H), 3.55-3.69 (m, 11H), 3.84-3.90 (m, 2H), 4.50 (s, 2H), 4.63 (t, J=3.6 Hz, 1H), 7.27-7.36 (m, 5H).

[00946] Step 3: Preparation of 3-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)propan-1-ol

[00947] A mixture of 2-(2-(2-(3-(benzyloxy)propoxy)ethoxy)ethoxy)tetrahydro-2H-pyran (2.4 g, 7.09 mmol) and palladium on carbon (10%, 240 mg ) in methanol (30 ml) was stirred at room temperature for 1 hour under a hydrogen atmosphere (hydrogen balloon). Palladium on carbon was removed by filtration and washed with methanol (10 ml). The combined filtrate was concentrated under reduced pressure to give 3-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)propan-1-ol (1.53 g, 92%) as gray oil.

[00948] Step 4: Preparation of 3-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)propanal

[00949] A mixture of 3-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)propan-1-ol (1.53 g, 6.16 mmol) and Dessa-periodinane Martin (5.2 g, 12.32 mmol) in dichloromethane (15 ml) was stirred at room temperature for 1 hour. The mixture was concentrated and the residue was washed with hexane (10 ml). The mixture was filtered and the filtrate was concentrated under reduced pressure to give 3-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)propanal (1.3 g, 85%) as colorless oils

[00950] Step 5: Preparation of tert-butyl (E)-5-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)pent-2-enoate

[00951] Mixture of 3-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)propanal (600 mg, 2.44 mmol) and tert-butyl 2-(triphenylphosphoranylidene) acetate (918 mg, 2.44 mmol) in dichloromethane (6 ml) was stirred at room temperature for 2 hours. The mixture was concentrated and the residue was purified by flash chromatography on silica gel (eluting with 13-20% ethyl acetate in hexane) to give (E)-tert-butyl5-(2-(2-((tetrahydro-2H-pyran)-2-yl )oxy)ethoxy)ethoxy)pent-2-enoate (420 mg, 50%) as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.48 (s, 9H), 1.50-1.62 (m, 4H), 1.69-1.76 (m, 1H), 1.79 -1.87 (m, 1H), 2.44-2.49 (m, 2H), 3.48-3.53 (m, 1H), 3.57-3.70 (m, 9H), 3 ,84-3.90 (m, 2H), 4.63 (t, J=3.6 Hz, 1H), 5.78-5.82 (m, 1H), 6.81-6.88 (m , 1H).

[00952] Step 6: Preparation of tert-butyl (E)-5-(2-(2-hydroxyethoxy)ethoxy)pent-2-enoate

[00953] Mixture of (E)-tert-butyl-5-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)pent-2-enoate (420 mg, 1.22 mmol) and pyridine-1-ium-4-methylbenzenesulfonate (154 mg, 0.61 mmol) in methanol (5 ml) were stirred at 50°C for 6 hours. The mixture was concentrated and the residue was partitioned between water (20 ml) and ethyl acetate (10 ml). The organic layer was collected, washed with brine (10 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give (E)-tert-butyl 5-(2-(2-hydroxyethoxy)ethoxy)pent-2-enoate (275 mg, 86%) as a colorless oil.

[00954] Step 7: Preparation of tert-butyl (E)-5-(2-(2-tosyloxy)ethoxy)ethoxy)pent-2-enoate

[00955] To a solution of (E)-tert-butyl 5-(2-(2-hydroxyethoxy)ethoxy)pent-2-enoate (275 mg, 1.05 mmol), triethylamine (215 mg, 2.1 mmol) and N,N-dimethylpyridin-4-amine (13 mg, 0.1 mmol) in dichloromethane (3 ml) was added with 4-toluenesulfonyl chloride (212 mg, 1.1 mmol) at 0°C. The mixture was stirred at room temperature for 3 hours. The mixture was diluted with dichloromethane (20 ml), washed with water (10 ml), brine (20 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue, which was purified by flash chromatography on silica gel (eluting at 18-25 % ethyl acetate in hexane) to obtain (E)-tert-butyl 5-(2-(2-(tosyloxy)ethoxy)ethoxy)pent-2-enoate (240 mg, 55%) as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.47 (s, 9H), 2.41-2.47 (m, 5H), 3.52-3.59 (m, 6H), 3.68 -3.70 (m, 2H), 4.15-4.18 (m, 2H), 5.77-5.81 (m, 1H), 6.81-6.84 (m, 1H), 7 .34 (d, J=8.0 Hz, 2H), 7.80 (d, J=8.4 Hz, 2H).

[00956] Step 8: Preparation of tert-butyl (E)-5-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy) ethoxy)ethoxy)pent-2-enoate

[00957] A mixture of (E)-tert-butyl 5-(2-(2-(tosyloxy)ethoxy)ethoxy)pent-2-enoate (240 mg, 0.58 mmol), 2-(2,6-dioxopiperidine- 3-yl)-5-hydroxyisoindoline-1,3-dione (159 mg, 0.58 mmol) and potassium carbonate (160 mg, 1.16 mmol) in dry N,N-dimethylformamide (3 ml) were stirred at 50° C for 16 hours. The mixture was partitioned between ethyl acetate (20 ml) and water (30 ml). The organic layer was collected, washed with brine (20 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by silica gel flash chromatography (eluting with 1-2% methanol in dichloromethane) to give (E) -tert-butyl 5-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)pent-2-enoate ( 200 mg, 67%) as a colorless oil. LC/MS (ESI) m/z: 517.40 [M+1] ⁺ .

[00958] Step 9: Preparation of (E)-5-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy )pent-2-enoic acid

[00959] To a solution of (E)-tert-butyl-5-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy )ethoxy)pent-2-enoate (100 mg, 0.19 mmol) in dichloromethane (2 ml) was added 2,2,2-trifluoroacetic acid (0.5 ml) at room temperature. The mixture was stirred at room temperature for 30 minutes. The mixture was concentrated and the residue was purified by preparative TLC (10% methanol in dichloromethane) to give (E)-5-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3 -dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)pent-2-enoic acid (64 mg, 72%) as a colorless oil. LC/MS (ESI) m/z: 461.10 [M+1] ⁺ .

[00960] Step 10: Preparation of (E)-5-(2-(2-((5-(4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-4- yl)piperazin-1-yl)-5-oxopent-3-en-1-yl)oxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione

[00961] To a solution of (E)-5-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)pent -2-enoic acid (42 mg, 0.09 mmol), 4-(6-chloro-8-fluoro-4-(piperazin-1-yl)quinazolin-7-yl)naphthalene-2-ol (40 mg, 0.1 mmol) and N-ethyl-N-isopropylpropan-2-amine (47 mg, 0.36 mmol) in dry N,N-dimethylformamide (1 ml) was added HATU (2-(7-aza-1H-benzotriazole -1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) (69 mg, 0.18 mmol) at 0° C., the resulting mixture was allowed to warm to room temperature, and stirred at room temperature for 15 minutes. The mixture was partitioned between ethyl acetate (10 ml) and water (10 ml). The organic layer was collected, washed with brine (20 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue, which was purified by HPLC to give (E)-5-(2-(2-((5-( 4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazin-1-yl)-5-oxopent-3-en-1-yl)oxy)ethoxy )ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (22 mg, 28%) as a white solid. LC/MS (ESI) m/z: 851.20 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 2.37-2.48 (m, 3H), 2.54-2.65 (m, 2H), 2.83-2.93 (m, 1H), 3.54-3.94 (m, 16H), 4.25-4.40 (m, 2H), 5.09-5.13 (m, 1H), 6.53-6.58 ( m, 1H), 6.70-6.77 (m, 1H), 7.11-7.46 (m, 7H), 7.82 (d, J=8.0 Hz, 2H), 8.08 (s, 1H), 8.71 (s, 1H), 10.06 (s, 1H), 11.10 (s, 1H).

[00962] Example of the synthesis of 4-(6-chloro-8-fluoro-2-(((S)-17-((2S,4R)-4-hydroxy-2-(((S)-1-(4- (4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-4,18,18-trimethyl-3,15-dioxo-7,10,13-trioxa-4,16-diazanonadecyl) amino)-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazine-1-carboxamide (representative compound 183)

[00963] Step 1: Preparation of tert-butyl-4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-2-((3-methoxy-3-oxopropyl)amino)quinazoline- 4-yl)piperazine-1-carboxylate

[00964] A mixture of tert-butyl 4-[7-bromo-6-chloro-8-fluoro-2-[(3-methoxy-3-oxopropyl)amino]quinazolin-4-yl]piperazine-1-carboxylate (1, 00 g, 1.83 mmol, 1 eq.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene-2-ol (593 mg, 2.19 mmol, 1.2 eq.), potassium phosphate (1.5 M, 3.7 mL, 3 eq.) and [2-(2-aminophenyl)phenyl]palladium (1+); dicyclohexyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane; methanesulfonate (155 mg, 0.18 mmol, 0.1 eq.) in tetrahydrofuran (10 ml) was degassed and purged with nitrogen gas 3 times, and then the mixture was stirred at 65°C for 12 hours under an atmosphere of nitrogen gas. The reaction mixture was diluted with water (50 ml), then extracted with ethyl acetate (50 ml×2). The combined organic layers were washed with brine (100 ml×1), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 5/1 to 1/1) to give tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl) )-2-[(3-methoxy-3-oxo-propyl)amino]quinazolin-4-yl]piperazine-1-carboxylate (780 mg, 1.28 mmol, 70% yield) as a brown solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.73 (d, J=8.4 Hz, 1H), 7.64 (d, J=1.6 Hz, 1H), 7.40 (t, J=7.6 Hz, 1H), 7.34-7.28 (m, 2H), 7.24-7.17 (m, 1H), 7.12 (d, J=2.0 Hz, 1H ), 5.86 (brs, 1H), 3.90-3.72 (m, 6H), 3.71-3.60 (m, 7H), 2.69 (t, J=6.4 Hz, 2H), 1.52 (s, 9H).

[00965] Step 2: Preparation of methyl-3-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl)amino) propanoate

[00966] To a solution of tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[(3-methoxy-3-oxopropyl)amino]quinazolin-4-yl ]piperazine-1-carboxylate (780 mg, 1.28 mmol, 1 eq.) in dichloromethane (5 ml) was added hydrochloric acid in dioxane (4 M, 5 ml). The mixture was stirred at 25°C for 15 minutes. The reaction mixture was concentrated under reduced pressure to obtain the compound methyl 3-[[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]amino] propanoate (700 mg, hydrochloride) as a yellow solid. LC/MS (ESI) m/z: 510.2 [M+1] ⁺ .

[00967] Step 3: Preparation of methyl-3-((4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl) amino)propanoate

[00968] To a solution of methyl 3-[[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]propanoate (650 mg, 1.19 mmol, 1 equiv., hydrochloride) and triethylamine (361 mg, 3.57 mmol, 0.5 ml, 3 equiv.) in dichloromethane (10 ml) was added a solution of isocyanato(trimethyl)silane (178 mg, 1.55 mmol, 1.3 equiv) in dichloromethane (1.5 ml). The mixture was stirred at 20°C for 0.5 hours. The reaction mixture was concentrated under reduced pressure to obtain the compound methyl 3-[[4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl ]amino]propanoate (700 mg) as a yellow solid. LC/MS (ESI) m/z: 553.2 [M+1] ⁺ .

[00969] Step 4: Preparation of 3-((4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino) propanoic acid

[00970] To a solution of methyl 3-[[4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]propanoate (700 mg, 1.27 mmol, 1 eq.) in tetrahydrofuran (10 ml) and water (1 ml) was added lithium hydroxide monohydrate (133 mg, 3.16 mmol, 2.5 eq.). The mixture was stirred at 20°C for 0.5 hours. The reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran. The residue was acidified to pH=3 with hydrochloric acid (1 M), and upon addition of hydrochloric acid some precipitate formed. The resulting mixture was filtered and the filter cake was evaporated to dryness to obtain the compound 3-[[4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin- 2-yl]amino]propanoic acid (640 mg, 1.19 mmol, 94% yield) as a brown solid. LC/MS (ESI) m/z: 539.1 [M+1] ⁺ .

[00971] Step 5: Preparation of 4-(6-chloro-8-fluoro-2-(((S)-17-((2S,4R)-4-hydroxy-2-(((S)-1-( 4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-4,18,18-trimethyl-3,15-dioxo-7,10,13-trioxa-4,16- diazanonedecyl)amino)-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazin-1-carboxamide

[00972] To a solution of (2S,4R)-1-((S)-15-(tert-butyl)-13-oxo-5,8,11-trioxa-2,14-diazahexadecane-16-oil)-4 -hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (70 mg, 0.93 mmol), 3-((4-( 4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)propanoic acid (40 mg, 0.074 mmol) and N-ethyl- N-isopropylpropan-2-amine (48 mg, 0.37 mmol) in dry N,N-dimethylformamide (1 ml) was added HATU (2-(7-aza-1H-benzotriazol-1-yl)-1,1, 3,3-Tetramethyluronium hexafluorophosphate) (72 mg, 0.18 mmol) at 0° C., the resulting mixture was allowed to warm to room temperature and stirred at room temperature for 10 minutes. The mixture was partitioned between ethyl acetate (10 ml) and water (10 ml). The organic layer was collected, washed with brine (10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by preparative TLC (10% methanol in dichloromethane) to give 4-(6-chloro-8- fluoro-2-(((S)-17-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl )pyrrolidine-1-carbonyl)-4,18,18-trimethyl-3,15-dioxo-7,10,13-trioxa-4,16-diazanonadecyl)amino)-7-(3-hydroxynaphthalene-1-yl) quinazolin-4-yl)piperazin-1-carboxamide (40 mg, 36% in two stages) as a light yellow solid. LC/MS (ESI) m/z: 1190.40 [M+23] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 0.92 (s, 9H), 1.17-1.26 (m, 3H), 1.31-1.39 (m, 3H), 1 .74-1.81 (m, 1H), 1.99-2.07 (m, 1H), 2.45 (s, 3H), 2.57-2.70 (m, 2H), 2.74 -2.92 (m, 2H), 3.01 (s, 1H), 3.39-3.62 (m, 20H), 3.86-3.98 (m, 2H), 4.22-4 .34 (m, 1H), 4.42-4.58 (m, 2H), 4.84-4.94 (m, 2H), 5.12 (s, 1H), 6.08 (s, 2H ), 7.04-7.43 (m, 11H), 7.73-7.86 (m, 2H), 8.42 (d, J=7.6 Hz, 1H), 8.97 (s, 1H), 9.97 (s, 1H).

[00973] Synthesis example of (2S,4R)-1-((2S)-21-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1 -yl)quinazolin-2-yl)amino)-2-(tert-butyl)-18-methyl-4,19-dioxo-6,9,12,15-tetraoxa-3,18-diazagenicosanoyl)-4-hydroxy -N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (representative compound 194)

[00974] Step 1: Preparation of 3-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2 -yl)amino)propanoic acid

[00975] To a solution of tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[(3-methoxy-3-oxo-propyl)amino]quinazoline-4 -yl]piperazine-1-carboxylate (7.02 g, 11.51 mmol, 1 eq.) in tetrahydrofuran (25 ml), methanol (25 ml) and water (50 ml) was added lithium hydroxide monohydrate (4.83 g , 115.07 mmol, 10 equiv), the mixture was stirred at 15°C for 2 hours. The mixture was poured into water (10 ml), the pH level was adjusted to 3 using 1 M hydrochloric acid. The aqueous phase was extracted with ethyl acetate (150 ml x 3). The combined organic phase was washed with brine (20 ml x 2), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The compound 3-[[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]propanoic acid ( 6.96 g) as a yellow solid. LC/MS (ESI) m/z: 596.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 2.19 (br s, 1H), 10.06 (s, 1H), 7.85-7.75 (m, 2H), 7.62- 7.35 (m, 2H), 7.28 (d, J=2.2 Hz, 1H), 7.23 (d, J=3.7 Hz, 2H), 7.07 (d, J=2 .2 Hz, 1H), 3.80-3.47 (m, 10H), 2.56 (br s, 2H), 1.45 (s, 9H).

[00976] Step 2: Preparation of tert-butyl-4-(6-chloro-8-fluoro-2-(((S)-20-((2S,4R)-4-hydroxy-2-(((S) -1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-4,21,21-trimethyl-3,18-dioxo-7,10,13,16- tetraoxa-4,19-diazadocosyl)amino)-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazin-1-carboxylate

[00977] To a solution of (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy ]ethoxy]acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (130 mg, 0. 18 mmol, 1 equiv., hydrochloride), 3-[[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2 -yl]amino]propanoic acid (106 mg, 0.18 mmol, 1 eq.) and N,N-diisopropylethylamine (69 mg, 0.54 mmol, 0.09 ml, 3 eq.) in N,N-dimethylformamide (3 ml) 1-hydroxybenzotriazole (29 mg, 0.21 mmol, 1.2 eq.) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (41 mg, 0.21 mmol, 1.2 eq.) were added sequentially .) at 0°C, then stirred at 20°C for 2 hours. The reaction mixture was quenched with water (30 ml) and then extracted with ethyl acetate (20 ml x 3). The combined organic phase was washed with brine (10 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo to leave a residue. The residue was purified by preparative TLC (dichloromethane/methanol 10:1). We obtained tert-butyl 4-[6-chloro-8-fluoro-2-[[3-[2-[2-[2-[2-[2-[[(1S)-1-[(2S,4R) -4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl]amino]-2 -oxoethoxy]ethoxy]ethoxy]ethoxy]ethylmethylamino]-3-oxopropyl]amino]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (123 mg, 0.09 mmol, yield 50%, purity 92%) as an off-white solid. LC/MS (ESI) m/z: 635.6 [M/2+1] ⁺ .

[00978] Step 3: Preparation of (2S,4R)-1-((2S)-2-(tert-butyl)-21-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl )-4-(piperazin-1-yl)quinazolin-2-yl)amino)-18-methyl-4,19-dioxo-6,9,12,15-tetraoxa-3,18-diazagenicosanoyl)-4-hydroxy -N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[00979] To a solution of tert-butyl-4-[6-chloro-8-fluoro-2-[[3-[2-[2-[2-[2-[2-[[(1S)-1-[ (2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl ]amino]-2-oxoethoxy]ethoxy]ethoxy]ethoxy]ethylmethylamino]-3-oxopropyl]amino]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (170 mg, 0.13 mmol, 1 eq.) in dichloromethane (5 ml), trifluoroacetic acid (770 mg, 6.75 mmol, 0.5 ml, 50.45 eq.) was added, then stirred at 20°C for 2 hours. The reaction mixture was concentrated to obtain a residue. We obtained (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(3- hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]propanoylmethylamino]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethylbutanoyl]-4-hydroxy-N -[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (170 mg, 0.13 mmol, 98% yield, trifluoroacetic acid) as a yellow solid .

[00980] Step 4: Preparation of (2S,4R)-1-((2S)-21-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene -1-yl)quinazolin-2-yl)amino)-2-(tert-butyl)-18-methyl-4,19-dioxo-6,9,12,15-tetraoxa-3,18-diazagenicosanoyl)-4 -hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[00981] To a solution of (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[3-[[6-chloro-8-fluoro-7 -(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]propanoylmethylamino]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethylbutanoyl]-4 -hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (170 mg, 0.13 mmol, 1 eq., trifluoroacetic acid) and 2,6-dimethylpyridine (142 mg, 1.32 mmol, 0.15 ml, 10 eq.) in dichloromethane (15 ml) was added a solution of prop-2-enoyl chloride (12 mg, 0.13 mmol, 0.01 ml, 1 eq.) in dichloromethane (10 ml) at minus 65°C, then stirred at this temperature for 10 minutes. The reaction mixture was quenched with water, 10 ml, and then extracted with dichloromethane (10 ml x 3). The combined organic phase was washed with brine (10 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo to leave a residue. The residue was purified by semipreparative reverse phase HPLC to give (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[3-[[6-chloro- 8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]amino]propanoylmethylamino]ethoxy]ethoxy]ethoxy]ethoxy]acetyl ]amino]-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (38 mg, 0 .03 mmol, yield 22%, purity 98%, formate) as an off-white solid. LC/MS (ESI) m/z: 612.3 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.02 (br s, 1H), 8.98 (s, 1H), 8.45 (br d, J=7.7 Hz, 1H), 8.17 (s, 1H), 7.84-7.76 (m, 2H), 7.49-7.33 (m, 6H), 7.28-7.10 (m, 4H), 7. 03 (d, J=2.2 Hz, 1H), 6.85 (dd, J=10.5, 16.7 Hz, 1H), 6.26-6.11 (m, 1H), 5.81 -5.67 (m, 1H), 5.14 (br s, 1H), 4.89 (brt, J=7.2 Hz, 1H), 4.53 (d, J=9.5 Hz, 1H ), 4.43 (t, J=8.0 Hz, 1H), 4.27 (br s, 1H), 3.99-3.88 (m, 2H), 3.86-3.65 (m , 7H), 3.62-3.60 (m, 1H), 3.61-3.40 (m, 21H), 3.03-2.77 (m, 1H), 3.03-2.77 (m, 1H), 3.03-2.77 (m, 1H), 2.61 (br s, 1H), 2.44 (s, 3H), 2.10-1.98 (m, 1H) , 1.81-1.70 (m, 1H), 1.48-1.31 (m, 3H), 0.96-0.88 (m, 1H), 0.92 (s, 8H).

[00982] Synthesis example of (2S,4R)-1-((S)-18-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-((R)-2 -fluoro-6-hydroxyphenyl)quinazolin-2-yl)amino)-2-(tert-butyl)-15-methyl-4,16-dioxo-6,9,12-trioxa-3,15-diazaoctadecanoyl)-4 -Hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (representative compound 210)

[00983] Step 1: Preparation of tert-butyl-4-(6-chloro-8-fluoro-7-((R)-2-fluoro-6-hydroxyphenyl)-2-(((S)-17-(( 2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-4,18,18- trimethyl-3,15-dioxo-7,10,13-trioxa-4,16-diazanonadecyl)amino)quinazolin-4-yl)piperazine-1-carboxylate

[00984] To a mixture of (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]acetyl] amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (146 mg, 0.21 mmol, 1, 2 equiv., hydrochloride) and 3-[[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazolin-2-yl] amino]propanoic acid (100 mg, 0.18 mmol, 1 eq.) in N,N-dimethylformamide (8 ml) was added 1-hydroxybenzotriazole (36 mg, 0.27 mmol, 1.5 eq.), 1-( 3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (51 mg, 0.27 mmol, 1.5 eq.) and N,N-diisopropylethylamine (69 mg, 0.53 mmol, 0.1 ml, 3 eq.). The mixture was stirred at 15°C for 16 hours. The reaction mixture was quenched with water (20 ml), then diluted with dichloromethane (50 ml) and extracted with dichloromethane (50 ml×2). The combined organic layers were washed with brine (30 ml), dried, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol = 10:1) to give tert-butyl-4-[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-[[3-[ 2-[2-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidin-1-carbonyl]-2,2-dimethylpropyl]amino]-2-oxoethoxy]ethoxy]ethoxy]ethylmethylamino]-3-oxopropyl]amino]quinazolin-4-yl]piperazine-1 -carboxylate (150 mg, 0.13 mmol, 71% yield) as a yellow oil. LC/MS (ESI) m/z: 1193.2 [M+1] ⁺ .

[00985] Step 2: Preparation of (2S,4R)-1-((S)-2-(tert-butyl)-18-((6-chloro-8-fluoro-7-((R)-2-fluoro -6-hydroxyphenyl)-4-(piperazin-1-yl)quinazolin-2-yl)amino)-15-methyl-4,16-dioxo-6,9,12-trioxa-3,15-diazaoctadecanoyl)-4 -hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[00986] To a solution of tert-butyl-4-[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-[[3-[2-[2-[2-[2- [[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1 -carbonyl]-2,2-dimethylpropyl]amino]-2-oxoethoxy]ethoxy]ethoxy]ethylmethylamino]-3-oxopropyl]amino]quinazolin-4-yl]piperazine-1-carboxylate (150 mg, 0.13 mmol, 1 eq.) in dichloromethane (8 ml) was added trifluoroacetic acid (3.08 g, 26.97 mmol, 2.00 ml, 215 eq.). The mixture was stirred at 15°C for 20 minutes. The mixture was concentrated to give (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(2- fluoro-6-hydroxyphenyl)-4-piperazin-1-ylquinazolin-2-yl]amino]propanoylmethylamino]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[ (1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (150 mg, 0.12 mmol, 99% yield, trifluoroacetate) as a yellow oil. LC/MS (ESI) m/z: 1093.1 [M+1] ⁺ .

[00987] Step 3: Preparation of (2S,4R)-1-((S)-18-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-((R) -2-fluoro-6-hydroxyphenyl)quinazolin-2-yl)amino)-2-(tert-butyl)-15-methyl-4,16-dioxo-6,9,12-trioxa-3,15-diazaoctadecanoyl) -4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[00988] To a solution of (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(2 -fluoro-6-hydroxyphenyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]propanoylmethylamino]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethylbutanoyl]-4-hydroxy- N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (76 mg, 0.06 mmol, 1 eq., trifluoroacetate) and 2,6- lutidine (67 mg, 0.63 mmol, 10 eq.) in dichloromethane (10 ml), prop-2-enoyl chloride (5 mg, 0.06 mmol, 0.9 eq.) was added dropwise at minus 70°C for 10 minutes. The reaction mixture was quenched with water (20 ml), then diluted with dichloromethane (50 ml) and extracted with dichloromethane (50 ml x 2). The combined organic layers were washed with brine (30 ml), dried, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by semipreparative reverse phase HPLC to give (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[3-[[6-chloro-8-fluoro -7-(2-fluoro-6-hydroxyphenyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]amino]propanoylmethylamino]ethoxy]ethoxy]ethoxy]acetyl]amino]-3 ,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (30 mg, 0.03 mmol, yield 42%, purity 100%) as a white solid. LC/MS (ESI) m/z: 1147.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.22 (br s, 1H), 8.98 (s, 1H), 8.45 (br d, J = 7.8 Hz, 1H), 7.72 (s, 1H), 7.55-6.99 (m, 7H), 6.92-6.68 (m, 3H), 6.17 (dd, J = 2.4, 16.7 Hz, 1H), 5.79-5.63 (m, 1H), 5.14 (br s, 1H), 4.89 (br t, J = 6.4 Hz, 1H), 4.54 (d , J = 9.4 Hz, 1H), 4.44 (t, J = 8.0 Hz, 1H), 4.28 (br s, 1H), 3.99-3.90 (m, 2H), 3.80-3.49 (m, 24H), 3.06-2.82 (m, 3H), 2.75-2.58 (m, 2H), 2.45 (s, 3H), 2. 04(br d, J = 8.0 Hz, 1H), 1.81-1.72 (m, 1H), 1.48-1.32 (m, 3H), 0.93 (br d, J = 2.4 Hz, 9H).

[00989] Example of the synthesis of (2S,4R)-1-((S)-21-(((S)-4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2 -fluoro-6-hydroxphenyl)quinazolin-2-yl)amino)-2-(tert-butyl)-18-methyl-4,19-dioxo-6,9,12,15-tetraoxa-3,18-diazagenicosanoyl) -4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (representative compound 211)

[00990] Step 1: Preparation of tert-butyl-4-((S)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-(((S)-20-(( 2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-4,21,21- trimethyl-3,18-dioxo-7,10,13,16-tetraoxa-4,19-diazadocosyl)amino)quinazolin-4-yl)piperazine-1-carboxylate

[00991] Mixture of (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy] ethoxy]acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (150 mg, 0.20 mmol, 1 equiv., hydrochloride) and 3-[[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)quinazoline-2- yl]amino]propanoic acid (129 mg, 0.20 mmol, 1 eq.) in N,N-dimethylformamide (8 ml) was added 1-hydroxybenzotriazole (41 mg, 0.30 mmol, 1.5 eq.), N -(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (59 mg, 0.30 mmol, 1.5 eq.) and N,N-diisopropylethylamine (79 mg, 0.61 mmol, 0.10 ml, 3 eq.) . The mixture was stirred at 20°C for 12 hours. The mixture was diluted with water (25 ml). The mixture was then extracted with ethyl acetate (20 ml x 3). The combined organic layer was washed with water (30 ml×2) and brine (30 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol = 10:1). The compound tert-butyl 4-[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-[[3-[2-[2-[2-[2-[2-[ [(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidin-1- carbonyl]-2,2-dimethylpropyl]amino]-2-oxoethoxy]ethoxy]ethoxy]ethoxy]ethylmethylamino]-3-oxopropyl]amino]quinazolin-4-yl]piperazine-1-carboxylate (150 mg, 0.12 mmol, yield 58%) in the form of a light yellow oil. LC/MS (ESI) m/z: 1237.6 [M+1] ⁺ .

[00992] Step 2: Preparation of (2S,4R)-1-((S)-2-(tert-butyl)-21-(((S)-6-chloro-8-fluoro-7-(2-fluoro -6-hydroxyphenyl)-4-(piperazin-1-yl)quinazolin-2-yl)amino)-18-methyl-4,19-dioxo-6,9,12,15-tetraoxa-3,18-diazagenicosanoyl) -4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[00993] To a mixture of tert-butyl 4-[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-[[3-[2-[2-[2-[2-[ 2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine -1-carbonyl]-2,2-dimethylpropyl]amino]-2-oxoethoxy]ethoxy]ethoxy]ethoxy]ethylmethylamino]-3-oxo-propyl]amino]quinazolin-4-yl]piperazine-1-carboxylate (150 mg , 0.12 mmol, 1 eq.) in dichloromethane (4 ml) was added trifluoroacetic acid (1 ml). The mixture was stirred at 20°C for 1 hour. The mixture was concentrated under reduced pressure to give (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[3-[[6-chloro-8-fluoro -7-(2-fluoro-6-hydroxyphenyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]propanoylmethylamino]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethylbutanoyl] -4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (150 mg, trifluoroacetate) as a light yellow oil. LC/MS (ESI) m/z: 1237.6 [M+1] ⁺ .

[00994] Step 3: Preparation of (2S,4R)-1-((S)-21-(((S)-4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7- (2-fluoro-6-hydroxphenyl)quinazolin-2-yl)amino)-2-(tert-butyl)-18-methyl-4,19-dioxo-6,9,12,15-tetraoxa-3,18- diazagenicosanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[00995] To a mixture of (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[3-[[6-chloro-8-fluoro-7 -(2-fluoro-6-hydroxyphenyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]propanoylmethylamino]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethylbutanoyl]-4 -hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (75 mg, 0.06 mmol, 1 eq., trifluoroacetate) and 2 ,6-lutidine (64 mg, 0.60 mmol, 0.07 ml, 10 eq.) in dichloromethane (20 ml) was added dropwise with prop-2-enoyl chloride (5 mg, 0.06 mmol, 1 eq.) to dichloromethane (5 ml) at minus 75°C in a nitrogen atmosphere. The mixture was then stirred at minus 75°C for 20 minutes. The mixture was quenched with water (20 ml). The mixture was then extracted with dichloromethane (20 ml x 3). The combined organic layer was washed with brine (30 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by semipreparative reverse phase HPLC. We obtained (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(2- fluoro-6-hydroxyphenyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]amino]propanoylmethylamino]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethylbutanoyl ]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (29.8 mg, 0.02 mmol, 39% yield , purity 98%, formate) as a white solid. LC/MS (ESI) m/z: 1213.3 [M+23] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 8.87 (s, 1H), 8.27 (s, 1H), 7.77 (s, 1H), 7.43-7.35 (m, 4H), 7.32-7.27 (m, 1H), 6.84-6.68 (m, 3H), 6.29-6.24 (m, 1H), 5.81-5.78 ( m, 1H), 5.01-4.98 (m, 1H), 4.69-4.66 (m, 1H), 4.60-4.55 (m, 1H, 4.11-3.98 (m, 2H), 3.90-3.82 (m, 7H), 3.75-3.72 (m, 4H), 3.66-3.51 (m, 17H), 3.15 (s , 1H), 2.96-2.94 (m, 2H), 2.86-2.83 (m, 1H), 2.78-2.74 (m, 1H), 2.47 (s, 3H ), 2.23-2.17 (m, 1H), 2.01-1.92 (m, 1H), 1.56-1.48 (m, 3H), 1.03-1.02 (m , 9H).

[00996] Example of the synthesis of (2S,4R)-1-((2S)-2-(tert-butyl)-14-((1-((2R)-2-((6-chloro-4-(4- (2,2-dihydroxyacetyl)piperazin-1-yl)-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)-4- oxo-6,9,12-trioxa-3-azatetradecanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide ( Typical connection 246)

[00997] Step 1: Preparation of tert-butyl 4-((11-oxo-3,6,9,12-tetraoxatetradecyl)oxy)piperidine-1-carboxylate

[00998] A solution of tert-butyl 4-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)piperidine-1-carboxylate (1.38 g, 4.14 mmol, 1.00 eq.) and dirhodium tetraacetate (II) (183 mg 0.41 mmol, 0.10 eq.) in dichloromethane (15 ml) was cooled to 0°C, then ethyl 2-diazoacetate (2.83 g, 24.84 mmol, 6. 00 eq.). The mixture was then stirred at 20°C for 8 hours. The reaction mixture was diluted with water (20 ml), then extracted with ethyl acetate (20 ml x 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 5/1 to 1/1) to obtain the desired product. The compound tert-butyl 4-((11-oxo-3,6,9,12-tetraoxatetradecyl)oxy)piperidine-1-carboxylate (1.36 g, 3.24 mmol, 78% yield) was obtained as a yellow oil. LC/MS (ESI) m/z 320.0 [M-99] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.21 (q, J = 7.2 Hz, 2H), 4.15 (s, 2H), 3.74-3.62 (m, 14H), 3.49-3.46 (m, 1H), 3.08-3.05 (m, 2H), 1.83 (d, J = 8.8 Hz, 2H), 1.53-1.49 ( m, 2H), 1.45 (s, 9H), 1.29 (t, J = 7.2 Hz, 3H).

[00999] Step 2: Preparation of 2-(2-(2-(2-((1-(tert-butoxycarbonyl)piperidin-4-yl)oxy)ethoxy)ethoxy)ethoxy)acetic acid

[001000] To a solution of tert-butyl-4-((11-oxo-3,6,9,12-tetraoxatetradecyl)oxy)piperidine-1-carboxylate (600 mg, 1.43 mmol, 1.00 eq.) in to a mixture of tetrahydrofuran (5 ml) and water (2 ml) lithium hydroxide hydrate (150 mg, 3.58 mmol, 2.50 eq.) was added. The mixture was stirred at 20°C for 0.5 hours. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was acidified to pH 3 with hydrochloric acid (1 M), then extracted with ethyl acetate (20 ml x 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The compound 2-(2-(2-(2-((1-(tert-butoxycarbonyl)piperidin-4-yl)oxy)ethoxy)ethoxy)ethoxy)acetic acid (550 mg) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.15 (s, 2H), 3.82-3.63 (m, 14H), 3.56-3.44 (m, 1H), 3.11 -2.95 (m, 2H), 1.91-1.80 (m, 2H), 1.58-1.48 (m, 2H), 1.45 (s, 9H).

[001001] Step 3: Preparation of tert-butyl 4-(((S)-13-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5 -yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-14,14-dimethyl-11-oxo-3,6,9-trioxa-12-azapentadecyl)oxy)piperidine-1-carboxylate

[001002] To a solution of 2-(2-(2-(2-((1-(tert-butoxycarbonyl)piperidin-4-yl)oxy)ethoxy)ethoxy)ethoxy)acetic acid (250 mg, 0.64 mmol, 1.00 equiv.) and (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4- 1-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (184 mg, 0.96 mmol, 1.50 eq.) and hydroxybenzotriazole (129 mg, 0.96 mmol, 1.50 eq.) and diisopropylethylamine (248 mg, 1.92 mmol, 3.00 eq. ). The reaction mixture was stirred at 25°C for 10 hours. The reaction mixture was diluted with water (20 ml), acidified to pH = 3 with hydrochloric acid (1 M), and then extracted with dichloromethane (20 ml x 2). The combined organic layers were washed with saturated sodium bicarbonate (20 ml), dried over sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate/methanol = 1/0 to 10/1) to obtain the desired product. The compound tert-butyl 4-(((S)-13-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)) was obtained ethyl)carbamoyl)pyrrolidine-1-carbonyl)-14,14-dimethyl-11-oxo-3,6,9-trioxa-12-azapentadecyl)oxy)piperidine-1-carboxylate (427 mg, 0.52 mmol, yield 82%) in the form of a colorless oil. LC/MS (ESI) m/z: 818.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.68 (s, 1H), 7.52-7.31 (m, 6H), 5.30 (s, 1H), 5.16-5.02 (m, 1H), 4.75 (t, J = 7.6 Hz, 1H), 4.59-4.46 (m, 2H), 4.15-4.09 (m, 1H), 3, 87-3.55 (m, 15H), 3.52-3.43 (m, 1H), 3.28 (s, 1H), 3.12-2.98 (m, 2H), 2.61- 2.49 (m, 4H), 2.11-2.05 (m, 1H), 1.89-1.78 (m, 5H), 1.51-1.42 (m, 12H), 1, 07 (s, 9H).

[001003] Step 4: Preparation of (2S,4R)-1-((S)-2-(tert-butyl)-4-oxo-14-(piperidin-4-yloxy)-6,9,12-trioxa- 3-azatetradecanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001004] To a solution of tert-butyl-4-(((S)-13-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5) -yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-14,14-dimethyl-11-oxo-3,6,9-trioxa-12-azapentadecyl)oxy)piperidine-1-carboxylate (427 mg, 0 .52 mmol, 1.00 eq.) in dichloromethane (2 mL), hydrogen chloride/dioxane (4.0 M, 3.0 mL, 22.99 eq.) was added. The mixture was stirred at 25°C for 0.5 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue. The compound (2S,4R)-1-((S)-2-(tert-butyl)-4-oxo-14-(piperidin-4-yloxy)-6,9,12-trioxa-3-azatetradecane-1 was obtained -oil)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (400 mg, hydrochloride salt) as a white solid . LC/MS (ESI) m/z: 718.4 [M+1] ⁺ .

[001005] Step 5: Preparation of tert-butyl-4-(6-chloro-8-fluoro-2-(((R)-1-(4-(((S)-13-((2S,4R)- 4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-14,14-dimethyl-11-oxo-3 ,6,9-trioxa-12-azapentadecyl)oxy)piperidin-1-yl)propan-2-yl)oxy)-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazin-1-carboxylate

[001006] To a mixture of (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-[2-[2-(4-piperidyloxy)ethoxy]ethoxy]ethoxy] acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (78 mg, 0.1 mmol, 1.2 eq. hydrochloride) in methanol (2 ml) was added sodium sulfate (28 mg, 0.34 mmol, 4 eq.). The reaction mixture was stirred at 25°C for 20 minutes. A solution of tert-butyl-4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxoethoxy]quinazolin-4-yl] was added piperazine-1-carboxylate (50 mg, 0.09 mmol, 1 eq.) in dichloromethane (2 ml), then acetic acid (10 mg, 0.17 mmol, 2 eq.). The reaction mixture was cooled to 0°C and sodium cyanoborohydride (16 mg, 0.26 mmol, 3 eq.) was added. The reaction mixture was stirred at 25°C for 4 hours. Methanol (2 ml) was added to obtain the reaction mixture as a clear solution. The solution was purified by preparative TLC (10% methanol in dichloromethane) to give tert-butyl-4-[6-chloro-8-fluoro-2-[(1R)-2-[4-[2-[2-[2- [2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]]carbamoyl ]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl]amino]-2-oxoethoxy]ethoxy]ethoxy]ethoxy]-1-piperidyl]-1-methylethoxy]-7-(3-hydroxy-1-naphthyl)quinazoline -4-yl]piperazine-1-carboxylate (45 mg, 0.03 mmol, 38% yield, 94% purity) as a colorless gum. LC/MS (ESI) m/z: 1281.6 [M+1] ⁺ .

[001007] Step 6: Preparation of (2S,4R)-1-((2S)-2-(tert-butyl)-14-((1-((2R)-2-((6-chloro-8-fluoro -7-(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)-4-oxo-6,9,12 -trioxa-3-azatetradecanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001008] To a solution of tert-butyl-4-[6-chloro-8-fluoro-2-[(1R)-2-[4-[2-[2-[2-[2-[[(1S)- 1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2, 2-dimethylpropyl]amino]-2-oxoethoxy]ethoxy]ethoxy]ethoxy]-1-piperidyl]-1-methylethoxy]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (45 mg, 0.03 mmol, 1 eq.) in dichloromethane (2 ml) was added trifluoroacetic acid (308 mg, 2.70 mmol, 0.2 ml, 77.01 eq.). The reaction mixture was stirred at 25°C for 1 hour. The reaction solution was concentrated in vacuo to give (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[[1-[(2R)-2-[6-chloro -8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]ethoxy]ethoxy]acetyl]amino] -3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (45 mg, 0.03 mol , yield 98%, trifluoroacetate) in the form of a colorless gum.

[001009] Step 7: Preparation of (2S,4R)-1-((2S)-2-(tert-butyl)-14-((1-((2R)-2-((6-chloro-4-( 4-(2,2-dihydroxyacetyl)piperazin-1-yl)-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)- 4-oxo-6,9,12-trioxa-3-azatetradecanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2- carboxamide

[001010] To a mixture of (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[[1-[(2R)-2-[6-chloro-8 -fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazin-1-ylquinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3 -dimethylbutanoyl]-4-hydroxy-N[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (85 mg, 0.07 mmol, 1 eq. hydrochloride) and 2,2-dihydroxyacetic acid (65 mg, 0.7 mmol, 10 eq.) in N,N-dimethylformamide (3 ml) O-(7-azabenzotriazol-1-yl)-N,N,N was added ',N'-tetramethyluronium exafluorophosphate (80 mg, 0.21 mmol, 3 eq.) and N,N-diisopropylethylamine (90 mg, 0.7 mmol, 0.1 ml, 10 eq.). The mixture was stirred at 25°C for 12 hours. The mixture was filtered. The filtrate was purified by preparative HPLC. The compound (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[[1-[(2R)-2-[6-chloro-4-[4 -(2,2-dihydroxyacetyl)piperazin-1-yl]-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]ethoxy]ethoxy ]acetyl]amino]-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (17, 5 mg, 0.01 mmol, yield 19%, purity 97.8%, formate) as a white solid. LC/MS (ESI) m/z: 1256.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.04 (s, 1H), 8.97 (s, 1H), 8.43-8.41 (d, J = 8.0 Hz, 1H ), 8.18 (s, 1H), 8.00 (s, 1H), 7.81-7.79 (d, J = 8.0 Hz, 1H), 7.58-7.26 (m, 7H), 7.24-7.14 (m, 2H), 7.09-7.01 (m, 1H), 6.43-6.30 (m, 1H), 5.52-5.03 ( m, 2H), 4.95-4.80 (m, 1H), 4.71-4.34 (m, 2H), 4.27 (s, 1H), 4.00-3.78 (m, 8H), 3.75-3.66 (m, 2H), 3.63-3.49 (m, 12H), 3.23-3.18 (m, 3H), 2.90-2.74 ( m, 2H), 2.44 (s, 4H), 2.18-1.98 (m, 5H), 1.81-1.66 (m, 3H), 1.40-1.18 (m, 8H), 0.92 (s, 9H).

[001011] Synthesis example of (2S,4R)-1-((S)-2-((E)-19-(3-(4-((4-chloro-2-hydroxy-5-(1-methylcyclopropyl) phenyl)glycyl)piperazin-1-yl)azetidin-1-yl)-19-oxo-3,6,9,12,15-pentaoxanonedec-17-enamido)-3,3-dimethylbutanoyl)-4-hydroxy-N -(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (representative compound 251)

[001012] Step 1: Preparation of tert-butyl 17-hydroxy-3,6,9,12,15-pentaoxaheptadecanoate

[001013] To a cold stirred solution of 3,6,9,12-tetraoxatetradecane-1,14-diol (3 eq.) in anhydrous N,N-dimethylformamide was added sodium hydride (60%, 1.2 eq.) portionwise at 0 °C in a nitrogen atmosphere. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 1 hour, then re-cooled to 0°C, tert-butyl 2-bromoacetate (1 eq.) was added in portions, the resulting mixture was allowed to warm to room temperature and stirred at room temperature temperature for 2 hours. The reaction was carefully quenched with water while cooling with ice water and extracted with methylene dichloride. The combined organic phase was washed with brine, dried over anhydrous magnesium sulfate and evaporated to dryness under reduced pressure. The crude product was purified by silica gel chromatography (eluent with 1-6% methanol in methylene dichloride) to give the desired product, tert-butyl 17-hydroxy-3,6,9,12,15-pentaoxaheptadecane-1-oate as a light yellow oil. yield 29%. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.48 (s, 9H), 3.60-3.73 (m, 20H), 4.02 (s, 2H).

[001014] Step 2: Preparation of tert-butyl 17-oxo-3,6,9,12,15-pentaoxaheptadecanoate

[001015] To a solution of tert-butyl 17-hydroxy-3,6,9,12,15-pentaoxaheptadecanoate (1 g, 3.52 mmol) in CH ₃ CN (30 ml) was added IBX (1.4 g, 4, 87 mmol). The mixture was stirred and refluxed for 5 hours under N ₂ atmosphere. The reaction mixture was cooled to room temperature and then filtered through celite. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EA = 5/1 to EA) to give the title compound (875 mg, 2.5 mmol, 71% yield).

[001016] Step 2: Preparation of 1-(tert-butyl) 19-methyl (E)-3,6,9,12,15-pentaoxanonedec-17-enedioate

[001017] To a solution of tert-butyl 17-oxo-3,6,9,12,15-pentaoxaheptadecanoate (875 mg, 2.5 mmol) and methyl 2-(dimethoxyphosphoryl) acetate (844 mg, 4.64 mmol) in THF (10 ml) was added DBU (1.1 g, 6.96 mmol). The mixture was stirred at room temperature for 5 hours under N ₂ atmosphere. The reaction mixture was quenched with water (20 ml). The organic phase was washed with brine. The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE/EA 30/1 to 1/1) to give the title compound (224 mg, 0.55 mmol, 22% yield).

[001018] Step 3: Preparation of (E)-3-oxo-2,7,10,13,16,19-hexaoxagenicos-4-ene-21-oic acid

[001019] To a solution of 1-(tert-butyl) 19-methyl (E)-3,6,9,12,15-pentaoxanonedec-17-enedioate (224 mg, 0.55 mmol) in DCM (5 ml) was added TFA (1 ml). The mixture was stirred at room temperature for 1 hour. The solution was concentrated under reduced pressure to obtain the product (E)-3-oxo-2,7,10,13,16,19-hexaoxagenicos-4-ene-21-oic acid (240 mg).

[001020] Step 4: Preparation of methyl (S,E)-3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1- carbonyl)-2,2-dimethyl-5-oxo-7,10,13,16,19-pentaoxa-4-azatricos-21-ene-23-oate

[001021] To a solution of (E)-3-oxo-2,7,10,13,16,19-hexaoxagenicos-4-ene-21-oic acid (240 mg) in DCM (10 ml) was added (2S,4R )-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride (257 mg, 0.55 mmol), TEA (154 mg, 1.52 mmol) and HATU (289 mg, 0.76 mmol). The mixture was stirred at room temperature for 1 hour. It was then quenched with H ₂ O (10 ml). The mixture was extracted with DCM. The combined organic layers were washed with brine. The organic phase was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM/MeOH = 50/1 to 20/1) to give methyl (S,E)-3-((2S,4R)-4-hydroxy-2-((4-( 4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)-2,2-dimethyl-5-oxo-7,10,13,16,19-pentaoxa-4-azatricos-21-en-23 -oate (206 mg, 0.27 mmol).

[001022] Step 5: Preparation of methyl (S,E)-3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1- carbonyl)-2,2-dimethyl-5-oxo-7,10,13,16,19-pentaoxa-4-azatricos-21-ene-23-oic acid

[001023] To a solution of methyl (S,E)-3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl) -2,2-dimethyl-5-oxo-7,10,13,16,19-pentaoxa-4-azatricos-21-ene-23-oate (206 mg, 0.27 mmol) in THF/water (5 ml /5 ml) NaOH (33 mg, 0.81 mmol) was added. The mixture was stirred at room temperature for 1 hour. The pH of the mixture was adjusted to 9 using aqueous HCl (1 M). The mixture was concentrated under reduced pressure to give methyl (S,E)-3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1- carbonyl)-2,2-dimethyl-5-oxo-7,10,13,16,19-pentaoxa-4-azatricos-21-ene-23-oic acid (220 mg).

[001024] Step 6: Preparation of (2S,4R)-1-((S)-2-((E)-19-(3-(4-((4-chloro-2-hydroxy-5-(1- methylcyclopropyl)phenyl)glycyl)piperazin-1-yl)azetidin-1-yl)-19-oxo-3,6,9,12,15-pentaoxanonedec-17-enamido)-3,3-dimethylbutanoyl)-4-hydroxy -N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[001025] To a solution of methyl (S,E)-3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl) 1-( 4-(azetidin-3-yl)piperazin-1-yl)-2-((4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)amino)ethan-1-one hydrochloride (116 mg, 0 .28 mmol), TEA (84 mg, 0.84 mmol) and HATU (160 mg, 0.42 mmol). The resulting mixture was stirred at room temperature for 30 minutes. It was quenched with H ₂ O (10 ml) and extracted with ethyl acetate. The combined organic layers were washed with brine. The organic phase was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC to give (2S,4R)-1-((S)-2-((E)-19-(3-(4-((4-chloro-2-hydroxy-5-(1-methylcyclopropyl )phenyl)glycyl)piperazin-1-yl)azetidin-1-yl)-19-oxo-3,6,9,12,15-pentaoxanonedec-17-enamido)-3,3-dimethylbutanoyl)-4-hydroxy- N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (15.5 mg, 0.014 mmol). ¹ H-NMR (400 MHz, CD ₃ OD) δ 8.87 (s, 1H), 8.25-8.26 (m, 1H), 7.41-7.47 (m, 4H), 6, 78-6.84 (m, 1H), 6.65 (s, 1H), 6.36 (s, 1H), 6.23-6.29 (m, 1H), 5.36-5.41 ( m, 1H), 4.69-4.71 (m, 1H), 4.50-4.59 (m, 3H), 4.29-4.38 (m, 2H), 4.18-4, 20 (m, 2H), 4.05-4.14 (m, 4H), 3.96 (s, 2H), 3.78-3.91 (m, 3H), 3.60-3.70 ( m, 20H), 3.24 (br, 1H), 2.48 (s, 3H), 2.37-2.45 (m, 4H), 2.20-2.25 (m, 1H), 2 .05-2.12 (m, 1H), 1.90 (s, 3H), 1.71-1.72 (m, 3H), 1.04 (s, 9H).

[001026] Example of the synthesis of (2S,4R)-1-((2S)-2-(2-((5-(4-(6-chloro-2-((3-(dimethylamino)-3-oxopropyl)amino )-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazin-1-yl)-5-oxopentyl)oxy)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy- N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (representative compound 254)

[001027] Step 1: Preparation of tert-butyl 17-hydroxy-3,6,9,12,15-pentaoxaheptadecanoate

[001028] Sodium hydroxide (10.67 g, 266.67 mmol, 11.08 eq.) was dissolved in water (16 ml) and then added to a solution of 3-benzyloxypropan-1-ol (4 g, 24.07 mmol , 3.81 ml, 1 eq.), tetra-n-butylammonium chloride (7.02 g, 25.27 mmol, 7.06 ml, 1.05 eq.) in dichloromethane (20 ml). Tert-Butyl 2-bromoacetate (18.78 g, 96.26 mmol, 14.22 mL, 4 eq.) was then added. The resulting mixture was stirred at 20°C for 16 hours. Water (80 ml) was added and the organic phase was separated. The aqueous phase was further extracted with dichloromethane (200 ml x 2). The combined organic phase was washed with brine (80 ml x 2), dried and concentrated. The residue was purified by chromatography on silica gel (petroleum ether/ethyl acetate 1/0 to 5/1) to give tert-butyl 2-(3-benzyloxypropoxy)acetate (1.0 g, 3.21 mmol, 13% yield, purity 90%) as a colorless oil and tert-butyl 2-(3-benzyloxypropoxy)acetate (6 g, 14.98 mmol, 62% yield, purity 70%) as a yellow oil.

[001029] Step 2: Preparation of tert-butyl 2-(3-hydroxypropoxy)acetate

[001030] To a solution of tert-butyl 2-(3-benzyloxypropoxy)acetate (6 g, 14.98 mmol, 1 eq.) in ethanol (10 ml) was added a palladium-activated carbon catalyst (0.5 g, 10% purity). and the mixture was degassed with hydrogen. The mixture was stirred at 15°C for 16 hours at a pressure of 0.1 MPa (15 psi). The mixture was then stirred at 40°C for 2 hours at a pressure of 0.35 MPa (50 psi). The mixture was filtered. The residue was purified by chromatography on silica gel (petroleum ether/ethyl acetate 5/1 to 1/1) to give tert-butyl 2-(3-hydroxypropoxy)acetate (1.1 g, 5.78 mmol, 39% yield). in the form of a colorless liquid. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 4.36 (t, J = 5.2 Hz, 1H), 3.92 (s, 2H), 3.53-3.38 (m, 4H ), 1.73-1.54 (m, 2H), 1.50-1.38 (s, 9H).

[001031] Step 3: Preparation of tert-butyl 2-(3-oxopropoxy)acetate

[001032] To a mixture of oxalyl chloride (1.20 g, 9.45 mmol, 0.83 mL, 3 eq.) in dichloromethane (8 mL), dimethyl sulfoxide (984 mg, 12.60 mmol, 0.98 mL, 4 eq.) in dichloromethane (4 ml) at minus 70°C and stirred for 0.5 hours at minus 70°C. tert-butyl 2-(3-hydroxypropoxy)acetate (600 mg, 3.15 mmol, 1 eq.) in dichloromethane (4 ml) was added dropwise and stirred at minus 70°C for 0.5 hour. Triethylamine (2.55 g, 25.20 mmol, 3.51 mL, 8 eq.) was added dropwise to the reaction mixture. The reaction mixture was then stirred at minus 70°C for 1 hour and 20°C for 1 hour. The reaction mixture was quenched with saturated ammonium chloride (20 ml) and then diluted with dichloromethane (50 ml) and extracted with dichloromethane (50 ml x 2). The combined organic layers were washed with brine (30 mL), dried, filtered and concentrated under reduced pressure to give tert-butyl 2-(3-oxopropoxy) acetate (500 mg, 2.66 mmol, 84.33% yield) as a yellow oil . ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 9.67 (t, J = 2.0 Hz, 1H), 8.75 (s, 1H), 4.05-3.91 (m, 2H ), 3.78 (t, J = 6.0 Hz, 2H), 2.62 (dt, J = 2.0, 6.0 Hz, 2H), 1.45-1.42 (s, 9H) .

[001033] Step 4: Preparation of methyl (E)-5-(2-(tert-butoxy)-2-oxoethoxy)pent-2-enoate

[001034] To a solution of tert-butyl 2-(3-oxopropoxy)acetate (500 mg, 2.66 mmol, 1 eq.) in dichloromethane (20 ml) was added methyl 2-(triphenylphosphanylidene) acetate (889 mg, 2 .66 mmol, 1 eq.). The mixture was stirred at 15°C for 16 hours. The mixture was concentrated as a residue. The residue was purified by chromatography on silica gel (petroleum ether/ethyl acetate 10/1 to 3/1) to give methyl (E)-5-(2-tert-butoxy-2-oxo-ethoxy)pent-2-enoate ( 350 mg, 1.43 mmol, 54% yield) as a yellow oil. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 6.91 (td, J = 6.8, 15.6 Hz, 1H), 5.97 (td, J = 1.6, 15.6 Hz , 1H), 4.06-3.89 (m, 2H), 3.65 (s, 3H), 3.60-3.50 (m, 2H), 2.45 (dq, J = 1.6 , 6.4 Hz, 2H), 1.43 (s, 9H).

[001035] Step 5: Preparation of (E)-2-((5-methoxy-5-oxopent-3-en-1-yl)oxy)acetic acid

[001036] To a solution of methyl (E)-5-(2-tert-butoxy-2-oxo-ethoxy)pent-2-enoate (300 mg, 1.23 mmol, 1 eq.) in dichloromethane (8 ml) was added trifluoroacetic acid (3.08 g, 27.01 mmol, 2 ml, 22 eq.). The mixture was stirred at 15°C for 1 hour. The mixture was concentrated to give 2-[(E)-5-methoxy-5-oxo-pent-3-enoxy]acetic acid (370 mg, 1.22 mmol, 99% yield, trifluoroacetate) as a yellow oil.

[001037] Step 6: Preparation of 2-((5-methoxy-5-oxopentyl)oxy)acetic acid

[001038] Palladium catalyst was added to a solution of 2-[(E)-5-methoxy-5-oxo-pent-3-enoxy]acetic acid (100 mg, 0.53 mmol, 1 eq.) in methanol (5 mL). on activated carbon (0.05 g, 10% purity) and the mixture was degassed with hydrogen. The entire mixture was stirred at 15°C for 16 hours at a pressure of 0.1 MPa (15 psi). The mixture was filtered to obtain a filtrate. The filtrate was concentrated to give 2-(5-methoxy-5-oxo-pentoxy)acetic acid (140 mg) as a yellow oil.

[001039] Step 7: Preparation of methyl 5-(2-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol- 5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)pentanoate

[001040] To a mixture of 2-(5-methoxy-5-oxo-pentoxy)acetic acid (140 mg, 0.74 mmol, 1 eq.) and (2S,4R)-1-[(2S)-2-amino -3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (327 mg, 0. 74 mmol, 1 eq.) in N,N,-dimethylformamide (8 ml), N,N-diisopropylethylamine (285 mg, 2.21 mmol, 0.38 ml, 3 eq.), 1-hydroxybenzotriazole (119 mg, 0.88 mmol, 1.2 eq.) and 3-(ethyliminomethyleneamino)-N,N-dimethylpropan-1-amine; hydrochloride (169 mg, 0.88 mmol, 1.2 eq.). The mixture was stirred at 15°C for 2 hours. The mixture was quenched with water (20 ml), the aqueous phase was extracted with ethyl acetate (50 ml x 2). The combined organic phase was washed with brine (30 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by semipreparative reverse phase HPLC to give methyl 5-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]pentanoate (120 mg, 0.19 mmol, 26% yield) in the form of a colorless oil. LC/MS (ESI) m/z: 617.5 [M+1] ⁺ .

[001041] Step 8: Preparation of 5-(2-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5 -yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)pentanoic acid

[001042] To a solution of methyl 5-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]pentanoate (120 mg, 0.19 mmol, 1 eq.) in tetrahydrofuran (3 ml), methanol (3 ml) and water (3 ml), lithium hydroxide monohydrate (251 mg, 5.98 mmol, 30.74 eq.) was added. The mixture was stirred at 0°C for 3 hours. The pH of the mixture was adjusted to 5 with hydrochloric acid (1 M) and the aqueous phase was extracted with ethyl acetate (150 ml). The combined organic phase was washed with brine (30 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo to give 5-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[ (1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]pentanoic acid (100 mg, 0.16 mmol, 85% yield) as a yellow oil.

[001043] Step 9: Preparation of tert-butyl (3-(dimethylamino)-3-oxopropyl)carbamate

[001044] To a solution of 2-cyano-N,N-dimethyl-acetamide (500 mg, 4.46 mmol, 1 eq.) in ethyl alcohol (10 ml) was added Raney Ni (100 mg, 1.17 mmol), and then ammonium hydroxide (1.82 g, 17.14 mmol, 2 ml, 33% purity), 3.84 eq.), di-tert-butyl dicarbonate (1.46 g, 6.69 mmol, 1.54 ml , 1.5 eq.), the mixture was stirred at 25°C for 10 hours under a hydrogen atmosphere [0.1 MPa (15 psi)]. The mixture was filtered and the filtrate was condensed to obtain the crude product. The crude product was purified by silica gel column chromatography (20-100% ethyl acetate in petroleum ether) to give the product tert-butyl N-[3-(dimethylamino)-3-oxo-propyl]carbamate (850 mg, 3.93 mmol, yield 88%) in the form of a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 5.43-5.27 (m, 1H), 3.41 (q, J = 5.6 Hz, 2H), 2.98 (s, 3H), 2.95 (s, 3H), 2.50 (t, J = 5.6 Hz, 2H), 1.42 (s, 9H).

[001045] Step 10: Preparation of 3-amino-N,N-dimethylpropanamide hydrochloride

[001046] To a solution of tert-butyl N-[3-(dimethylamino)-3-oxo-propyl]carbamate (850 mg, 3.93 mmol, 1 eq.) in dichloromethane (8 mL) was added hydrochloric acid/methanol (4 M, 8 ml, 8.14 eq.), the mixture was stirred at 25°C for 2 hours. The mixture was condensed to give 3-amino-N,N-dimethylpropanamide (580 mg, 3.80 mmol, 96.7% yield, hydrochloric acid) as a colorless oil. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.02 (br s, 3H), 2.98-2.90 (m, 5H), 2.83 (s, 3H), 2.68 ( t, J = 6.7 Hz, 2H).

[001047] Step 11: Preparation of tert-butyl 4-(7-bromo-6-chloro-2-((3-(dimethylamino)-3-oxopropyl)amino)-8-fluoroquinazolin-4-yl)piperazin-1- carboxylate

[001048] Diisopropylethylamine (810 mg, 6.27 mmol, 1.1 ml, 3.5 eq.), and then tert-butyl 4-(7-bromo-2,6-dichloro-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate (860 mg, 1.79 mmol, 1 eq.), the mixture was stirred at 95°C for 12 hours. The mixture was condensed to give a residue, the residue was dissolved in ethyl acetate (100 ml) and washed with water (100 ml), the organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was condensed to give the crude product. This crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether/dichloromethane = 5/1/1, 3/1/1, 2/1/1, 1/1/1) to give tert-butyl 4-[ 7-bromo-6-chloro-2-[[3-(dimethylamino)-3-oxo-propyl]amino]-8-fluoroquinazolin-4-yl]piperazine-1-carboxylate (750 mg, 1.34 mmol , yield 74%) as a brown solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.56 (d, J = 1.9 Hz, 1H), 5.92 (br t, J = 6.2 Hz, 1H), 3.84 (q , J = 6.2 Hz, 2H), 3.61 (br s, 8H), 3.03 (s, 3H), 2.97 (s, 3H), 2.67 (br s, 2H), 1 .50 (s, 9H).

[001049] Step 12: Preparation of tert-butyl 4-(6-chloro-2-((3-(dimethylamino)-3-oxopropyl)amino)-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazoline -4-yl)piperazine-1-carboxylate

[001050] To a solution of tert-butyl 4-[7-bromo-6-chloro-2-[[3-(dimethylamino)-3-oxo-propyl]amino]-8-fluoroquinazolin-4-yl]piperazin- 1-carboxylate (332 mg, 0.59 mmol, 1 eq.) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene-2-ol (160 mg , 0.59 mmol, 1 eq.) in dioxane (8 ml) and water (2 ml) added sodium carbonate (188 mg, 1.78 mmol, 3 eq.), tetrakis[triphenylphosphine]palladium(0) (68 mg , 0.06 mmol, 0.1 eq.), the mixture was stirred at 90°C under nitrogen atmosphere for 5 hours. Tetrakis[triphenylphosphine]palladium(0) (34.22 mg, 0.029 mmol, 0.05 eq) was charged into the reaction mixture and stirred at 90°C under nitrogen atmosphere for 3 hours. The mixture was diluted with water (50 ml) and extracted with ethyl acetate (50 ml, twice), the organic phase was dried with anhydrous sodium sulfate, filtered and the filtrate was condensed to give the crude product. This crude product was purified by preparative TLC (1/1 dichloromethane/ethyl acetate) to give tert-butyl 4-[6-chloro-2-[[3-(dimethylamino)-3-oxo-propyl]amino]-8- fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (100 mg, 0.16 mmol, 26.6% yield, 98.2% purity) as a brown solid substances. LC/MS (ESI) m/z: 623.1 [M+1] ⁺ .

[001051] Step 13: Preparation of 3-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl)amino)-N ,N-dimethylpropanamide

[001052] To a solution of tert-butyl 4-[6-chloro-2-[[3-(dimethylamino)-3-oxo-propyl]amino]-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline -4-yl]piperazine-1-carboxylate (100 mg, 0.16 mmol, 1 eq.) in dichloromethane (8 ml) was added trifluoroacetic acid (3.08 g, 27.01 mmol, 2 ml, 168.32 eq. .), the mixture was stirred at a temperature of 25°C for 1 hour. The mixture was concentrated to obtain the crude product. This crude product was diluted with saturated aqueous sodium bicarbonate (20 ml) and extracted with ethyl acetate (20 ml, three times), the organic phase was dried with anhydrous sodium sulfate, filtered and the filtrate was condensed to give 3-[[6-chloro-8-fluoro-7- (3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]-N,N-dimethylpropanamide (95 mg) as a brown solid. LC/MS (ESI) m/z: 523.1 [M+1] ⁺ .

[001053] Step 14: Preparation of (2S,4R)-1-((2S)-2-(2-((5-(4-(6-chloro-2-((3-(dimethylamino)-3-oxopropyl )amino)-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazin-1-yl)-5-oxopentyl)oxy)acetamido)-3,3-dimethylbutanoyl)-4- hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001054] To a mixture of 5-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl )phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]pentanoic acid (100 mg, 0.17 mmol, 1 eq.) and 3-[ [6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]-N,N-dimethylpropanamide (106 mg, 0 .17 mmol, 1 eq., trifluoroacetate) in N,N-dimethylformamide (4 ml) was added N,N-diisopropylethylamine (64 mg, 0.50 mmol, 3 eq.), 3-(ethyliminomethyleneamino)-N,N- dimethylpropan-1-amine; hydrochloride (38 mg, 0.20 mmol, 1.2 eq.) and 1-hydroxybenzotriazole (27 mg, 0.20 mmol, 1.2 eq.). The mixture was stirred at 15°C for 3 hours. The reaction mixture was quenched with water (20 ml) and then diluted with dichloromethane (50 ml) and extracted with dichloromethane (50 ml x 2). The combined organic layers were washed with brine (30 ml), dried over sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by semipreparative reverse phase HPLC to give (2S,4R)-1-[(2S)-2-[[2-[5-[4-[6-chloro-2-[[3-(dimethylamino)-3 -oxo-propyl]amino]-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazin-1-yl]-5-oxo-pentoxy]acetyl]amino]-3,3 -dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (9 mg, 0.01 mmol, yield 4.6%, purity 95%) as a white solid. LC/MS (ESI) m/z: 554.4 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.78-9.66 (s, 1H), 9.09-8.87 (s, 1H), 8.45 (d, J = 7, 6 Hz, 1H), 8.38 (s, 1H), 7.80 (br d, J = 6.0 Hz, 2H), 7.43 (br d, J = 8.0 Hz, 3H), 7 .38-7.31 (m, 3H), 7.27 (d, J = 2.0 Hz, 1H), 7.25-7.10 (m, 3H), 7.05 (d, J = 2 ,4 Hz, 1H), 5.15 (br s, 1H), 4.89 (br t, J = 7.2 Hz, 1H), 4.55 (d, J = 9.4 Hz, 1H), 4.46 (t, J = 8.0 Hz, 1H), 4.29 (br s, 1H), 3.97-3.89 (m, 2H), 3.71 (br s, 5H), 3 .59 (br s, 3H), 3.52 (br s, 5H), 2.98 (s, 3H), 2.80 (br s, 3H), 2.61 (br t, J = 6.8 Hz, 2H), 2.47-2.40 (m, 6H), 2.12-2.01 (m, 1H), 1.81-1.72 (m, 1H), 1.62 (br s , 4H), 1.49-1.35 (m, 3H), 0.94 (s, 9H).

[001055] Example of the synthesis of 3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N -(2-(2-(2-(3-(2-((S)-1-((S)-2-cyclohexyl-2-((S)-2-(methylamino)propanamido)acetyl)pyrrolidine- 2-yl)thiazol-4-carbonyl)phenoxy)ethoxy)ethoxy)ethyl)-N-methylpropanamide (typical compound 257)

[001056] Step 1: Preparation of 2-(trimethylsilyl)ethyl(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)(methyl)carbamate

[001057] To a stirred solution of 2-(2-(2-(methylamino)ethoxy)ethoxy)ethan-1-ol (1.8 g, 0.005 mol) and triethylamine (1.01 g, 0.01 mol) in dichloromethane (30 ml), 4-nitrophenyl (2-(trimethylsilyl)ethyl) carbonate (3.1 g, 0.02 mol) was added at 0°C. The resulting mixture was stirred at room temperature overnight. The reaction mixture was diluted with dichloromethane (20 ml), washed with water and brine (30 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue, which was purified by flash chromatography on silica gel (eluting with 20-30% ethyl acetate in hexane ) to give 2-(trimethylsilyl)ethyl(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)(methyl)carbamate (560 mg, 42%) as a colorless oil.

[001058] Step 2: Preparation of 2,2,7-trimethyl-6-oxo-5,10,13-trioxa-7-aza-2-silapentadecan-15-yl 4-methylbenzenesulfonate

[001059] To a stirred solution of 2-(trimethylsilyl)ethyl(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)(methyl)carbamate (560 mg, 1.43 mmol), triethylamine (436 mg, 4.3 mmol ) and N,N-dimethylpyridin-4-amine (17 mg, 0.14 mmol) in dichloromethane (10 ml), 4-toluenesulfonyl chloride (820 mg, 4.3 mmol) was added at 0°C. The resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to give a crude residue, which was purified by flash chromatography on silica gel (eluting with 20-40% ethyl acetate in hexane) to give 2,2,7-trimethyl-6-oxo-5,10,13-triox -7-aza-2-silapentadecan-15-yl 4-methylbenzenesulfonate (300 mg, 45%) as a white solid.

[001060] Step 3: Preparation of tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-oxo-2-((S)-2-(4-(3-((2, 2,7-trimethyl-6-oxo-5,10,13-trioxa-7-aza-2-silapentadecan-15-yl)oxy)benzoyl)thiazol-2-yl)pyrrolidin-1-yl)ethyl)amino) -1-oxopropan-2-yl)(methyl)carbamate

[001061] To a stirred solution of 2,2,7-trimethyl-6-oxo-5,10,13-trioxa-7-aza-2-silapentadecan-15-yl 4-methylbenzenesulfonate (115 mg, 0.24 mmol) and potassium carbonate (103 mg, 0.74 mmol) in N,N-dimethylformamide (5 ml) was added tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-((S)-2 -(4-(3-hydroxybenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate (100 mg, 0.16 mmol) . The resulting mixture was stirred at 50°C overnight. The reaction mixture was partitioned between ethyl acetate (20 ml) and water (10 ml). The organic layer was collected, and the aqueous layer was extracted with ethyl acetate (15 ml x 2). The combined organic layers were washed with brine (10 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (eluting with 3-5% methanol in dichloromethane) to give tert-butyl ((S )-1-(((S)-1-cyclohexyl-2-oxo-2-((S)-2-(4-(3-((2,2,7-trimethyl-6-oxo-5,10 ,13-trioxa-7-aza-2-silapentadecan-15-yl)oxy)benzoyl)thiazol-2-yl)pyrrolidin-1-yl)ethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate (120 mg, 49%) as a white solid.

[001062] Step 4: Preparation of tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-((S)-2-(4-(3-(2-(2-(2 -(methylamino)ethoxy)ethoxy)ethoxy)benzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate

[001063] To a stirred solution of tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-oxo-2-((S)-2-(4-(3-((2.2 ,7-trimethyl-6-oxo-5,10,13-trioxa-7-aza-2-silapentadecan-15-yl)oxy)benzoyl)thiazol-2-yl)pyrrolidin-1-yl)ethyl)amino)- 1-oxopropan-2-yl)(methyl)carbamate (120 mg, 0.13 mmol) in tetrahydrofuran (5 ml) was added tetrabutylammonium fluoride (80 mg, 0.30 mmol). The resulting mixture was stirred at room temperature for 4 hours. The reaction mixture was partitioned between ethyl acetate (20 ml) and water (10 ml). The organic layer was collected, and the aqueous layer was extracted with ethyl acetate (15 ml x 2). The combined organic layers were washed with brine (10 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (eluting with 3-5% methanol in dichloromethane) to give tert-butyl ((S )-1-(((S)-1-cyclohexyl-2-((S)-2-(4-(3-(2-(2-(2-(methylamino)ethoxy)ethoxy)ethoxy)benzoyl)thiazole -2-yl)pyrrolidin-1-yl)-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate (130 mg, 80%) as a white solid.

[001064] Step 5: Preparation of tert-butyl ((2S)-1-(((1S)-2-((2S)-2-(4-(3-(2-(2-(2-(3- ((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy) ethoxy)benzoyl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate

[001065] To a stirred solution of tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-((S)-2-(4-(3-(2-(2-(2- (methylamino)ethoxy)ethoxy)ethoxy)benzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate (130 mg, 0.14 mmol), (2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) (96 mg, 0.23 mmol) and N-ethyl-N-isopropylpropane-2 -amine (49 mg, 0.37 mmol) in N,N-dimethylformamide (5 ml) was added 3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3 -hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)propanoic acid (70 mg, 0.12 mmol). The resulting mixture was stirred at room temperature for 1 hour. The reaction mixture was partitioned between ethyl acetate (20 ml) and water (10 ml). The organic layer was collected, and the aqueous layer was extracted with ethyl acetate (15 ml x 2). The combined organic layers were washed with brine (10 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a residue, which was purified by preparative TLC (eluting with 10% methanol in dichloromethane) to give tert-butyl ((S)-1-(( (S)-2-((S)-2-(4-(3-(2-(2-(2-(3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8 -fluoro-7-((E)-3-hydroxy-1-(o-tolyl)prop-1-en-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)ethoxy) benzoyl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate (80 mg, 49%) as a white solid .

[001066] Step 6: Preparation of 3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino) -N-(2-(2-(2-(3-(2-((S)-1-((S)-2-cyclohexyl-2-((S)-2-(methylamino)propanamido)acetyl) pyrrolidin-2-yl)thiazol-4-carbonyl)phenoxy)ethoxy)ethoxy)ethyl)-N-methylpropanamide

[001067] Tert-Butyl solution ((S)-1-(((S)-2-((S)-2-(4-(3-(2-(2-(2-(3-((4 -(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-((E)-3-hydroxy-1-(o-tolyl)prop-1-en-1-yl)quinazolin-2 -yl)amino)-N-methylpropanamido)ethoxy)ethoxy)ethoxy)benzoyl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)amino)-1-oxopropan-2-yl) (methyl)carbamate (80 mg, 0.062 mmol) in 4 M hydrogen chloride in dioxane (2 ml) was stirred at room temperature for 20 minutes. The volatile components were evaporated under reduced pressure to obtain a residue, which was dissolved in aqueous sodium bicarbonate (1 N, 10 ml) and extracted with dichloromethane (20 ml). The combined organic layer was collected, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude residue, which was purified by preparative TLC (eluting with 10% methanol in chloromethane) to give 3-((4-(4-acryloylpiperazin-1-yl)- 6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-(2-(2-(2-(3-(2-((S)- 1-((S)-2-cyclohexyl-2-((S)-2-(methylamino)propanamido)acetyl)pyrrolidin-2-yl)thiazol-4-carbonyl)phenoxy)ethoxy)ethoxy)ethyl)-N- methylpropanamide (60 mg, 82%) as a white solid. LC/MS (ESI) m/z: 1175.5 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 0.86 (s, 1H), 1.06 (s, 6H), 1.16-1.28 (m, 5H), 1.62 (d , J = 1.2 Hz, 6H), 2.01 (s, 2H), 2.19 (s, 1H), 2.32 (s, 3H), 2.63 (d, J = 25.4 Hz , 2H), 2.80 (s, 1H), 2.99 (s, 1H), 3.38-3.60 (m, 11H), 3.61-3.91 (m, 12H), 4, 12 (dd, J = 1.4, 0.8 Hz, 2H), 4.47 (s, 1H), 5.32-5.43 (m, 1H), 5.73 (d, J = 10, 4 Hz, 1H), 6.16 (d, J = 16.8 Hz, 1H), 6.76-6.89 (m, 1H), 7.05 (d, J = 1.9 Hz, 1H) , 7.18-7.29 (m, 4H), 7.42 (s, 2H), 7.58-7.66 (m, 2H), 7.78 (s, 2H), 8.38-8 .26 (m, 1H), 8.47 (s, 1H), 10.01 (s, 1H).

[001068] Example of the synthesis of 3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-((S)-2-fluoro-6-hydroxyphenyl)quinazolin-2-yl) amino)-N-(2-(2-(2-((4-(2-((S)-1-((S)-2-cyclohexyl-2-((S)-2-(methylamino)propanamido )acetyl)pyrrolidin-2-yl)thiazol-4-yl)naphthalene-1-yl)oxy)ethoxy)ethoxy)ethyl)-N-methylpropanamide (typical compound 277)

[001069] Step 1: Preparation of tert-butyl ((S)-1-(((S)-2-((S)-2-(4-(4-(2-(2-(2-(3- ((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-((S)-2-fluoro-6-hydroxyphenyl)quinazolin-2-yl)amino)-N-methylpropanamido) ethoxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)amino)-1-oxopropan-2-l)(methyl)carbamate

[001070] To a mixture of 3-[[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxy-phenyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2- yl]amino]propanoic acid (50 mg, 96.54 µmol, 1 eq.) and tert-butyl N-[(1S)-2-[[(1S)-1-cyclohesyl-2-[(2S)-2 -[4-[4-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-2-oxo-ethyl]amino ]-1-methyl-2-oxo-ethyl]-N-methyl-carbamate (78.39 mg, 102.34 µmol, 1.06 eq.) in dimethylformamide (5 ml) was added diisopropylethylamine (49.91 mg, 386 .17 µmol, 67.26 µl, 4 eq.), hydroxybenzotriazole (26.09 mg, 193.08 µmol, 2 eq.) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (37.01 mg, 193 .08 µmol, 2 eq.) in one serving. The mixture was stirred at 20°C for 12 hours. The reaction mixture was washed with water (50 ml) and extracted with ethyl acetate (30 ml x 3), then the organic phase was concentrated in vacuo. The residue was purified by preparative HPLC to give the compound tert-butylN-[(1S)-2-[[(1S)-2-[(2S)-2-[4-[4-[2-[2-[2-[3 -[[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxy-phenyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]amino]propanoyl- methyl-amino]ethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-1-cyclohexyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl ]-N-methyl-carbamate (60 mg, 47.40 µmol, 49% yield) as a white solid. LC/MS (ESI) m/z: 633.3 [M/2+1] ⁺ .

[001071] Step 2: Preparation of 3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-((S)-2-fluoro-6-hydroxyphenyl)quinazoline-2- yl)amino)-N-(2-(2-(2-((4-(2-((S)-1-((S)-2-cyclohexyl-2-((S)-2-(methylamino )propanamido)acetyl)pyrrolidin-2-yl)thiazol-4-yl)naphthalene-1-yl)oxy)ethoxy)ethoxy)ethyl)-N-methylpropanamide

[001072] To a mixture of tert-butyl N-[(1S)-2-[[(1S)-2-[(2S)-2-[4-[4-[2-[2-[2-[3- [[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxy-phenyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]amino]propanoyl-methyl -amino]ethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-1-cyclohexyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl] -N-methyl-carbamate (60 mg, 47.40 µmol, 1 eq.) in dichloromethane (5 ml) was added trifluoroacetic acid (770.00 mg, 6.75 mmol, 0.5 ml, 142.48 eq.) one portion. The mixture was stirred at 20°C for 1 hour. The reaction mixture was concentrated under reduced pressure to remove the solvent to obtain a residue. The residue was purified by preparative HPLC to give the compound (2S)-N-[(1S)-2-[(2S)-2-[4-[4-[2-[2-[2-[3-[[6-chloro -8-fluoro-7-(2-fluoro-6-hydroxy-phenyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy] ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-1-cyclohexyl-2-oxo-ethyl]-2-(methylamino)propanamide (23.4 mg, 18.10 µmol, yield 38%, purity 99%), trifluoroacetic acid) as a white solid. LC/MS (ESI) m/z: 583.3 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 8.25 (br d, J = 7.3 Hz, 1H), 8.14-8.01 (m, 1H), 7.85-7.63 (m, 1H), 7.52 - 7.32 (m, 5H), 6.91-6.66 (m, 4H), 6.30 (br d, J = 16.4 Hz, 1H), 5 .83 (br d, J = 10.3 Hz, 1H), 5.51 (br d, J = 7.2 Hz, 1H), 4.67-4.58 (m, 1H), 4.28 ( br s, 2H), 4.16-3.51 (m, 26H), 3.16-3.03 (m, 1H), 2.97 (s, 2H), 2.86 (br s, 1H) , 2.75-2.65 (m, 3H), 2.46-2.07 (m, 4H), 1.95-1.61 (m, 6H), 1.51 (d, J = 7, 0 Hz, 3H), 1.31-1.06 (m, 5H).

[001073] Example of the synthesis of 4-(6-chloro-2-((3-((2-(2-(2-((4-(2-((S)-1-((S)-2-cyclohexyl -2-((S)-2-(methylamino)propanamido)acetyl)pyrrolidin-2-yl)thiazol-4-yl)naphthalene-1-yl)oxy)ethoxy)ethoxy)ethyl)(methyl)amino)-3 -oxopropyl)amino)-8-fluoro-7-((S)-2-fluoro-6-hydroxyphenyl)quinazolin-4-yl)piperazine-1-carboxamide (typical compound 287)

[001074] Step 1: Preparation of tert-butyl ((S)-1-(((S)-2-((S)-2-(4-(4-(2-(2-(2-(3- ((4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-((S)-2-fluoro-6-hydroxyphenyl)quinazolin-2-yl)amino)-N-methylpropanamido) ethoxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)amino)-1-oxopropan-2-l)(methyl)carbamate

[001075] To a mixture of tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-((S)-2-(4-(4-(2-(2-(2-( methylamino)ethoxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate (60, 45 mg, 78.91 µmol, 1 eq.), (S)-3-((4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6- hydroxyphenyl)quinazolin-2-yl)amino)propanoic acid (40 mg, 78.91 µmol, 1 eq.) and N-ethyl-N-isopropylpropan-2-amine (40.80 mg, 315.65 µmol, 54, 98 µl, 4 eq.) in dimethylformamide (5 ml) was added hydroxybenzotriazole (21.33 mg, 157.83 µmol, 2.0 eq.) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (30.26 mg, 157.83 µmol, 2 eq.) in one serving. The mixture was stirred at 20°C for 12 hours. The reaction mixture was washed with water (50 ml) and extracted with ethyl acetate (30 ml x 3), then the organic phase was concentrated in vacuo. The residue was purified by preparative thin layer chromatography (silica, ethyl acetate:methanol = 40:1). Tert-butyl compound ((S)-1-(((S)-2-((S)-2-(4-(4-(2-(2-(2-(3-((4-(4 -carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-((S)-2-fluoro-6-hydroxyphenyl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)ethoxy) naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate (60 mg, 40.16 µmol , yield 51%, purity 84%) was obtained as a white solid. LC/MS (ESI) m/z: 1254.8 [M+1] ⁺ .

[001076] Step 2: Preparation of 4-(6-chloro-2-((3-((2-(2-(2-((4-(2-((S)-1-((S)-2 -cyclohexyl-2-((S)-2-(methylamino)propanamido)acetyl)pyrrolidin-2-yl)thiazol-4-yl)naphthalene-1-yl)oxy)ethoxy)ethoxy)ethyl)(methyl)amino) -3-oxopropyl)amino)-8-fluoro-7-((S)-2-fluoro-6-hydroxyphenyl)quinazolin-4-yl)piperazine-1-carboxamide

[001077] To a solution of tert-butyl ((S)-1-(((S)-2-((S)-2-(4-(4-(2-(2-(2-(3-(( 4-(4-carbamoylpiperazin-1-yl)-6-chloro-8-fluoro-7-((S)-2-fluoro-6-hydroxyphenyl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy) ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate (50 mg, 39.84 µmol, 1 eq.) in dichloromethane (2 ml), trifluoroacetic acid (1.54 g, 13.51 mmol, 1000 µl, 338.97 eq.) was added. The mixture was stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC. Compound 4-(6-chloro-2-((3-((2-(2-(2-((4-(2-((S)-1-((S)-2-cyclohexyl-2-( (S)-2-(methylamino)propanamido)acetyl)pyrrolidin-2-yl)thiazol-4-yl)naphthalene-1-yl)oxy)ethoxy)ethoxy)ethyl)(methyl)amino)-3-oxopropyl)amino )-8-fluoro-7-((S)-2-fluoro-6-hydroxyphenyl)quinazolin-4-yl)piperazine-1-carboxamide (8.7 mg, 6.58 µmol, 16% yield, 96% purity ), trifluoroacetic acid) was obtained as a yellow solid. LC/MS (ESI) m/z: 1176.5 [M+23] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 8.27 (br d, J = 8.0 Hz, 1H), 8.09 (d, J = 8.4 Hz, 1H), 7.81- 7.67 (m, 1H), 7.50-7.37 (m, 5H), 6.88-6.76 (m, 3H), 5.55 (br d, J = 8.0 Hz, 1H ), 4.65 (d, J = 6.8 Hz, 1H), 4.31 (br s, 2H), 4.06-3.58 (m, 25H), 2.99 (s, 2H), 2.89 (br s, 1H), 2.70 (s, 3H), 2.47 (br s, 1H), 2.29 (br d, J = 6.4 Hz, 2H), 2.17 ( br s, 1H), 1.96-1.62 (m, 7H), 1.53 (d, J = 6.8 Hz, 3H), 1.37-1.10 (m, 6H).

[001078] Example of the synthesis of (2R,3S,4R,5S)-N-(4-((2-(2-(2-(3-((4-(4-acryloylpiperazin-1-yl)-6-chloro -8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)ethyl)carbamoyl)-2-methoxyphenyl)-3-(3-chloro- 2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamide (representative compound 302)

[001079] Step 1: Preparation of tert-butyl 4-(6-chloro-2-((1-(4-((2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4 -(4-chloro-2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamido)-3-methoxyphenyl)-11-methyl-1,12-dioxo-5,8-dioxa-2,11-diazatetradecane -14-yl)amino)-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazine-1-carboxylate

[001080] To a mixture of (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-( 2,2-dimethylpropyl)-N-[2-methoxy-4-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethylcarbamoyl]phenyl]pyrrolidine-2-carboxamide (140 mg, 0.16 mmol, 1 equiv., trifluoroacetate) and 3-[[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl] amino]propanoic acid (95 mg, 0.16 mmol, 1 eq.) in N,N-dimethylformamide (8 ml) was added 1-hydroxybenzotriazole (32 mg, 0.24 mmol, 1.5 eq.), N-( 3-dimethylamino1phopyl)-N-ethylcarbodiimide hydrochloride (46 mg, 0.24 mmol, 1.5 eq.) and N,N-diisopropylethylamine (103 mg, 0.80 mmol, 0.14 ml, 5 eq.). The mixture was stirred at 20°C for 15 hours. The mixture was diluted with water (25 ml), extracted with ethyl acetate (25 ml × 3). The combined organic layer was washed with water (30 ml×2) and brine (30 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative thin layer chromatography (dichloromethane:methanol = 10:1). tert-butyl 4-[6-chloro-2-[[3-[2-[2-[2-[[4-[[(2R,3S,4R,5S)-3-(3-chloro-2- fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3-methoxy-benzoyl]amino]ethoxy ]ethoxy]ethyl-methyl-amino]-3-oxo-propyl]amino]-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (150 mg, 0 .11 mmol, 70% yield) was obtained as a light yellow solid. LC/MS (ESI) m/z: 1339.4 [M+1] ⁺ .

[001081] Step 2: Preparation of (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-(4-((2- (2-(2-(3-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl)amino)-N -methylpropanamido)ethoxy)ethoxy)ethyl)carbamoyl)-2-methoxyphenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamide

[001082] To a mixture of tert-butyl 4-[6-chloro-2-[[3-[2-[2-[2-[[4-[[(2R,3S,4R,5S)-3-(3 -chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3-methoxy- benzoyl]amino]ethoxy]ethoxy]ethyl-methyl-amino]-3-oxo-propyl]amino]-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (150 mg, 0.11 mmol, 1 eq.) in dichloromethane (5 ml) was added trifluoroacetic acid (1 ml). The mixture was stirred at 20°C for 0.5 hour. The mixture was concentrated under reduced pressure to obtain the product. (2R,3S,4R,5S)-N-[4-[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4 -piperazin-1-yl-quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethylcarbamoyl]-2-methoxy-phenyl]-3-(3-chloro-2-fluorophenyl)-4 -(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide (150 mg, trifluoroacetate) was obtained as a light yellow oil. LC/MS (ESI) m/z: 1239.4 [M+1] ⁺ .

[001083] Step 3: Preparation of (2R,3S,4R,5S)-N-(4-((2-(2-(2-(3-((4-(4-acryloylpiperazin-1-yl)-6 -chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)ethyl)carbamoyl)-2-methoxyphenyl)-3-(3- chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamide

[001084] To a mixture of (2R,3S,4R,5S)-N-[4-[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(3-hydroxy-1 -naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethylcarbamoyl]-2-methoxy-phenyl]-3-(3-chloro-2-fluoro -phenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide (150 mg, 0.11 mmol, 1 eq., trifluoroacetate ) and 2,6-lutidine (118 mg, 1.11 mmol, 0.13 ml, 10 eq.) in dichloromethane (20 ml) was added prop-2-enoyl chloride (10 mg, 0.11 mmol, 0.009 ml, 1 eq.) in dichloromethane (2 ml) at minus 78°C in a nitrogen atmosphere. The mixture was stirred at a temperature of minus 78°C for 0.5 hours. The mixture was quenched with water (20 ml). It was extracted with dichloromethane (20 ml x 2). The combined organic layer was washed with brine (20 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC. (2R,3S,4R,5S)-N-[4-[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4 -(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethylcarbamoyl]-2-methoxy-phenyl]-3-(3-chloro-2 -fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide (40.1 mg, 0.03 mmol, yield 27%, purity 97%) was obtained as an off-white solid. LC/MS (ESI) m/z: 1293.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.39 (s, 1H), 9.99 (s, 1H), 8.45-8.42 (m, 1H), 8.30-8 .29 (m, 2H), 7.79-7.77 (m, 2H), 7.74-7.71 (m, 1H), 7.58-7.48 (m, 5H), 7.41 . 734 (m, 4H), 7.26 (s, 1H), 7.25-7.21 (m, 2H), 7.04 (s, 1H), 6.86-6.80 (m, 1H) , 6.18-6.14 (m, 1H), 5.73-5.71 (m, 1H), 4.59 (s, 2H), 4.27-4.41 (m, 1H), 3 .97-3.90 (m, 4H), 3.80-3.71 (m, 10H), 3.52-3.45 (m, 12H), 2.98-2.79 (m, 3H) , 2.62-2.60 (m, 1H), 1.66-1.60 (m, 1H), 1.28-1.24 (m, 1H), 0.96 (s, 9H).

[001085] Example of the synthesis of (2R,3S,4R,5S)-N-(4-((2-(2-(2-(3-((4-(4-acetylpiperazin-1-yl)-6-chloro -8-fluoro-7-((S)-2-fluoro-6-hydroxyphenyl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)ethyl)carbamoyl)-2-methoxyphenyl)-3-( 3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamide (typical compound 319)

[001086] Step 1: Preparation of tert-butyl 4-(6-chloro-2-((1-(4-((2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4 -(4-chloro-2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamido)-3-methoxyphenyl)-11-methyl-1,12-dioxo-5,8-dioxa-2,11-diazatetradecane -14-yl)amino)-8-fluoro-7-((S)-2-fluoro-6-hydroxyphenyl)quinazolin-4-yl)piperazine-1-carboxylate

[001087] To a mixture of (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-( 2,2-dimethylpropyl)-N-[2-methoxy-4-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethylcarbamoyl]phenyl]pyrrolidine-2-carboxamide (175 mg, 0.2 mmol, 1 equiv., trifluoroacetate) and 4-[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxy-phenyl)quinazolin-2-yl ]butanoic acid (112 mg, 0.2 mmol, 1 eq.) in N,N-dimethylformamide (8 ml) was added 1-hydroxybenzotriazole (54 mg, 0.40 mmol, 2 eq.), N-(3-dimethylaminopropyl )-N-ethylcarbodiimide hydrochloride (76 mg, 0.40 mmol, 2 eq.) and N,N-diisopropylethylamine (129 mg, 1.00 mmol, 0.17 ml, 5 eq.). The mixture was stirred at 25°C for 12 hours. The mixture was diluted with water (25 ml). It was then extracted with ethyl acetate (25 ml x 3). The combined organic layer was washed with water (30 ml×2) and brine (30 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative thin layer chromatography (dichloromethane:methanol = 10:1). Tert-butyl compound 4-[6-chloro-2-[[3-[2-[2-[2-[[4-[[(2R,3S,4R,5S)-3-(3-chloro-2 -fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3-methoxy-benzoyl]amino] ethoxy]ethoxy]ethyl-methyl-amino]-3-oxo-propyl]amino]-8-fluoro-7-(2-fluoro-6-hydroxy-phenyl)quinazolin-4-yl]piperazin-1-carboxylate (180 mg, 0.14 mmol, 68% yield) was obtained as a light yellow solid. LC/MS (ESI) m/z: 1307.4 [M+1] ⁺ .

[001088] Step 2: Preparation of (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-(4-((2- (2-(2-(3-((6-chloro-8-fluoro-7-((S)-2-fluoro-6-hydroxyphenyl)-4-(piperazin-1-yl)quinazolin-2-yl) amino)-N-methylpropanamido)ethoxy)ethoxy)ethyl)carbamoyl)-2-methoxyphenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamide

[001089] To a mixture of tert-butyl 4-[6-chloro-2-[[3-[2-[2-[2-[[4-[[(2R,3S,4R,5S)-3-(3 -chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3-methoxy- benzoyl]amino]ethoxy]ethoxy]ethyl-methyl-amino]-3-oxo-propyl]amino]-8-fluoro-7-(2-fluoro-6-hydroxy-phenyl)quinazolin-4-yl]piperazine-1 -carboxylate (180 mg, 0.14 mmol, 1 eq.) in dichloromethane (5 ml) was added trifluoroacetic acid (1 ml). The mixture was stirred at 25°C for 0.5 hour. The mixture was concentrated under reduced pressure to obtain the product. (2R,3S,4R,5S)-N-[4-[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl) -4-piperazin-1-yl-quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethylcarbamoyl]-2-methoxy-phenyl]-3-(3-chloro-2-fluorophenyl) -4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide (180 mg, trifluoroacetate) was obtained as a light yellow oil. LC/MS (ESI) m/z: 1207.3 [M+1] ⁺ .

[001090] Step 3: Preparation of (2R,3S,4R,5S)-N-(4-((2-(2-(2-(3-((4-(4-acetylpiperazin-1-yl)-6 -chloro-8-fluoro-7-((S)-2-fluoro-6-hydroxyphenyl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)ethyl)carbamoyl)-2-methoxyphenyl)-3 -(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamide

[001091] To a mixture of (2R,3S,4R,5S)-N-[4-[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(2-fluoro-6 -hydroxy-phenyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethylcarbamoyl]-2-methoxy-phenyl]-3-(3-chloro-2 -fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide (90 mg, 0.07 mmol, 1 eq. , trifluoroacetate) and 2,6-lutidine (73 mg, 0.68 mmol, 0.08 ml, 10 eq.) in dichloromethane (20 ml) was added acetyl chloride (5 mg, 0.07 mmol, 0.004 ml, 1 eq. ) in dichloromethane (5 ml) at minus 78°C in a nitrogen atmosphere. The mixture was stirred at minus 78°C for 20 minutes. The mixture was quenched with water (20 ml). It was then extracted with dichloromethane (20 ml x 2). The combined organic layer was washed with brine (30 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC. Compound (2R,3S,4R,5S)-N-[4-[2-[2-[2-[3-[[4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro- 7-(2-fluoro-6-hydroxy-phenyl)quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethylcarbamoyl]-2-methoxy-phenyl]-3-(3-chloro-2- fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide (51.1 mg, 0.04 mmol, yield 59 %, purity 99%) was obtained as a white solid. LC/MS (ESI) m/z: 1249.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.49-10.39 (m, 2H), 8.48-8.44 (m, 1H), 8.31 (dd, J = 2, 0, 8.4 Hz, 1H), 8.02-8.01 (m, 2H), 7.74-7.71 (m, 1H), 7.58-7.45 (m, 4H), 7 ,38-7.33 (m, 4H), 6.88-6.79 (m, 2H), 4.58 (s, 2H), 4.16-4.12 (m, 3H), 3.96 -3.90 (m, 7H), 3.55-3.37 (m, 17H), 2.96-2.84 (m, 3H), 2.71-2.65 (m, 2H), 2 .04-2.03 (m, 3H), 1.67-1.61 (m, 1H), 1.28-1.24 (m, 1H), 0.96 (s, 9H).

[001092] Example of the synthesis of (2R,3S,4R,5S)-N-(4-((2-(2-(2-(3-((4-(4-acetylpiperazin-1-yl)-6-chloro -8-fluoro-7-((R)-2-fluoro-6-hydroxyphenyl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)ethyl)carbamoyl)-2-methoxyphenyl)-3-( 3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamide (typical compound 320)

[001093] Step 1: Preparation of tert-butyl 4-(6-chloro-2-((1-(4-((2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4 -(4-chloro-2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamido)-3-methoxyphenyl)-11-methyl-1,12-dioxo-5,8-dioxa-2,11-diazatetradecane -14-yl)amino)-8-fluoro-7-((R)-2-fluoro-6-hydroxyphenyl)quinazolin-4-yl)piperazine-1-carboxylate

[001094] To a mixture of (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-( 2,2-dimethylpropyl)-N-[2-methoxy-4-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethylcarbamoyl]phenyl]pyrrolidine-2-carboxamide (175 mg, 0.20 mmol, 1 equiv., trifluoroacetate) and 4-[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(2-fluoro-6-hydroxy-phenyl)quinazolin-2-yl ]butanoic acid (112 mg, 0.20 mmol, 1 eq.) in N,N-dimethylformamide (8 ml) was added 1-hydroxybenzotriazole (54 mg, 0.40 μmol, 2 eq.), N-(3-dimethylaminopropyl )-N-ethylcarbodiimide hydrochloride (76 mg, 0.40 mmol, 2 eq.) and N,N-diisopropylethylamine (129 mg, 1.00 mmol, 0.17 ml, 5 eq.). The mixture was stirred at 25°C for 12 hours. The mixture was diluted with water (20 ml). It was then extracted with ethyl acetate (20 ml x 3). The combined organic layer was washed with water (30 ml×2) and brine (30 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative thin layer chromatography (dichloromethane:methanol = 10:1). Tert-butyl compound 4-[6-chloro-2-[[3-[2-[2-[2-[[4-[[(2R,3S,4R,5S)-3-(3-chloro-2 -fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3-methoxy-benzoyl]amino] ethoxy]ethoxy]ethyl-methyl-amino]-3-oxo-propyl]amino]-8-fluoro-7-(2-fluoro-6-hydroxy-phenyl)quinazolin-4-yl]piperazin-1-carboxylate (140 mg, 0.11 mmol, 53% yield) was obtained as a light yellow solid. LC/MS (ESI) m/z: 1307.4 [M+1] ⁺ .

[001095] Step 2: Preparation of (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-N-(4-((2- (2-(2-(3-((6-chloro-8-fluoro-7-((R)-2-fluoro-6-hydroxyphenyl)-4-(piperazin-1-yl)quinazolin-2-yl) amino)-N-methylpropanamido)ethoxy)ethoxy)ethyl)carbamoyl)-2-methoxyphenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamide

[001096] To a mixture of tert-butyl 4-[6-chloro-2-[[3-[2-[2-[2-[[4-[[(2R,3S,4R,5S)-3-(3 -chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3-methoxy- benzoyl]amino]ethoxy]ethoxy]ethyl-methyl-amino]-3-oxo-propyl]amino]-8-fluoro-7-(2-fluoro-6-hydroxy-phenyl)quinazolin-4-yl]piperazine-1 -carboxylate (140 mg, 0.11 mmol, 1 eq.) in dichloromethane (5 ml) was added trifluoroacetic acid (1 ml). The mixture was stirred at 25°C for 0.5 hour. The mixture was concentrated under reduced pressure to obtain the product. (2R,3S,4R,5S)-N-[4-[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl) -4-piperazin-1-yl-quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethylcarbamoyl]-2-methoxy-phenyl]-3-(3-chloro-2-fluorophenyl) -4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide (140 mg, trifluoroacetate) was obtained as a light yellow oil. LC/MS (ESI) m/z: 1207.4 [M+1] ⁺ .

[001097] Step 3: Preparation of (2R,3S,4R,5S)-N-(4-((2-(2-(2-(3-((4-(4-acetylpiperazin-1-yl)-6 -chloro-8-fluoro-7-((R)-2-fluoro-6-hydroxyphenyl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)ethyl)carbamoyl)-2-methoxyphenyl)-3 -(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamide

[001098] To a mixture of (2R,3S,4R,5S)-N-[4-[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(2-fluoro-6 -hydroxy-phenyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethylcarbamoyl]-2-methoxy-phenyl]-3-(3-chloro-2 -fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide (70 mg, 0.05 mmol, 1 eq. , trifluoroacetate) and 2,6-lutidine (56 mg, 0.53 mmol, 0.06 ml, 10 eq.) in dichloromethane (20 ml) was added acetyl chloride (4 mg, 0.05 mmol, 0.004 ml, 1 eq. ) in dichloromethane (5 ml) at minus 78°C in a nitrogen atmosphere. The mixture was stirred at minus 78°C for 20 minutes. The mixture was quenched with water (20 ml). It was then extracted with dichloromethane (20 ml x 2). The combined organic layer was washed with brine (30 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC. Compound (2R,3S,4R,5S)-N-[4-[2-[2-[2-[3-[[4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro- 7-(2-fluoro-6-hydroxy-phenyl)quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethylcarbamoyl]-2-methoxy-phenyl]-3-(3-chloro-2- fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide (30.1 mg, 0.02 mmol, yield 45 %, purity 99%) was obtained as a white solid. LC/MS (ESI) m/z: 1249.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.50-10.39 (m, 2H), 8.47-8.45 (m, 1H), 8.31 (dd, J = 2, 0, 8.4 Hz, 1H), 8.02-8.00 (m, 2H), 7.74-7.71 (m, 1H), 7.58-7.46 (m, 4H), 7 .40-7.32 (m, 4H), 6.88-6.79 (m, 2H), 4.58 (s, 2H), 4.16-4.12 (m, 3H), 3.93 -3.90 (m, 7H), 3.54-3.37 (m, 17H), 2.96-2.84 (m, 3H), 2.71-2.65 (m, 2H), 2 .04-2.03 (m, 3H), 1.67-1.61 (m, 1H), 1.28-1.24 (m, 1H), 0.96 (s, 9H).

[001099] Example of the synthesis of (2S,4R)-N-(2-(2-(2-(3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-( 3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)^-methylpropanamido)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((R)- 3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide (representative compound 339)

[001100] Step 1: Preparation of (2S,4R)-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)-N-(2-(2- (2-(methylamino)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[001101] Mixture of tert-butyl N-[2-[2-[2-[[[(2S,4R)-4-hydroxy-1-[(2R)-3-methyl-2-(3-methylisoxazole-5 -yl)butanoyl]pyrrolidin-2-carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]ethoxy]ethyl]-N-methyl-carbamate (90 mg, 0.12 mmol, 1 equiv .) in hydrochloric acid/dioxane (4 M, 10 ml, 311 eq.) was degassed and purged with nitrogen 3 times, and then the mixture was stirred at 25°C for 0.5 hour under nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure to obtain a residue. Compound (2S,4R)-4-hydroxy-N-[[2-[2-[2-(methylamino)ethoxy]ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]-1-[ (2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (75 mg, 0.11 mmol, 91% yield, hydrochloride) was obtained as a colorless oil. LC/MS (ESI) m/z: 600.4 [M+1] ⁺ .

[001102] Step 2: Preparation of tert-butyl 4-(6-chloro-8-fluoro-2-((3-((2-(2-(2-(((2S,4R)-4-hydroxy-1 -((R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)ethyl) methyl)amino)-3-oxopropyl)amino)-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazine-1-carboxylate

[001103] Mixture of (2S,4R)-4-hydroxy-N-[[2-[2-[2-(methylamino)ethoxy]ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]- 1-[(2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (75 mg, 0.11 mmol, 1 eq., hydrochloride), 3-[[4 -(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]propanoic acid (70 mg, 0.11 mmol , 1 eq.), 1-hydroxybenzotriazole (23 mg, 0.17 mmol, 1.5 eq.), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (33 mg, 0.17 mmol, 1.5 eq. .) and N,N-diisopropylethylamine (61 mg, 0.47 mmol, 4 eq.) in N,N-dimethylformamide (1 ml) were degassed and purged with nitrogen 3 times, and then the mixture was stirred at 25°C for 12 hours in a nitrogen atmosphere. The reaction mixture was quenched by adding 10 ml of water, and extracted with ethyl acetate (20 ml x 3). The combined organic layers were washed with brine (10 ml x 3), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (9% methanol in dichloromethane). tert-butyl 4-[6-chloro-8-fluoro-2-[[3-[2-[2-[2-[[[(2S,4R)-4-hydroxy-1-[(2R)-3 -methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]ethoxy]ethyl-methyl-amino]-3 -oxo-propyl]amino]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (75 mg, 0.06 mmol, 50% yield, 93% purity) was obtained as yellow oil. LC/MS (ESI) m/z: 590.1 [M/2+1] ⁺ .

[001104] Step 3: Preparation of (2S,4R)-N-(2-(2-(2-(3-((6-chloro-8fluoro-7-(3-hydroxynaphthalene-1-yl)-4-( piperazin-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((R)-3 -methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide

[001105] Mixture of tert-butyl 4-[6-chloro-8-fluoro-2-[[3-[2-[2-[2-[[[(2S,4R)-4-hydroxy-1-[( 2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]ethoxy]ethyl-methyl- amino]-3-oxo-propyl]amino]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (80 mg, 0.06 mmol, 1 eq.) and trifluoroacetic acid (821 mg, 7.20 mmol, 106.05 eq.) in dichloromethane (5 ml) was degassed and purged with nitrogen 3 times, and then the mixture was stirred at 25°C for 1 hour under nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure to give (2S,4R)-N-[[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl) -4-piperazin-1-yl-quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]-4-hydroxy-1- [(2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (75 mg, 0.06 mmol, 92% yield), trifluoroacetate) as a yellow oil.

[001106] Step 4: Preparation of (2S,4R)-N-(2-(2-(2-(3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7 -(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-(( R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide

[001107] To a solution of (2S,4R)-N-[[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4- piperazin-1-yl-quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]-4-hydroxy-1-[(2R )-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (75 mg, 0.06 mmol, 1 eq., trifluoroacetate) and 2,6-lutidine (67 mg, 0 .62 mmol, 0.073 ml, 10 eq.) in dichloromethane (20 ml), a solution of prop-2-enoyl chloride (6 mg, 0.06 mmol, 0.05 ml, 1 eq.) in dichloromethane (1 ml) was added dropwise at minus 78°C in a nitrogen atmosphere. The reaction solution was stirred at minus 78°C for 0.5 hours. The reaction mixture was quenched by adding 10 ml of water, and extracted with ethyl acetate (20 ml x 3). The combined organic layers were washed with brine (10 ml x 3), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by semipreparative reverse phase HPLC. Compound 2S,4R)-N-[[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop- 2-enoylpiperazin-1-yl)quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]-4-hydroxy-1-[ (2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (45 mg, 0.04 mol, 62% yield, 99% purity) was obtained as a white solid. LC/MS (ESI) m/z: 566.5 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 9.98 (br s, 1H), 8.97 (s, 1H), 8.30-8.21 (m, 1H), 7.79 ( br d, J = 7.9 Hz, 2H), 7.52-7.34 (m, 1H), 7.32-6.94 (m, 8H), 6.89-6.79 (m, 1H ), 6.24-6.14 (m, 2H), 5.74 (br d, J = 10.6 Hz, 1H), 5.11 (br s, 1H), 4.55-4.15 (m, 6H), 3.93-3.76 (m, 11H), 3.66-3.49 (m, 6H), 3.03-2.70 (m, 4H), 2.65-2.55 ( m, 4H), 2.47-2.39 (m, 3H), 2.30-2.03 (m, 1H), 2.15-2.12 (m, 2H), 2.07-2.00 ( m, 1H), 1.91 (br dd, J = 5.1, 7.6 Hz, 1H), 0.97 (d, J = 6.6 Hz, 2H), 0.80 (d, J = 6.7 Hz, 2H), 0.68 (br d, J = 6.5 Hz, 1H), 0.56 (br d, J = 6.4 Hz, 1H).

[001108] Example of the synthesis of (2S,4R)-N-(2-(2-(2-(3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-( 3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((S) -3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide (typical compound 340)

[001109] Step 1: Preparation of (2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)-N-(2-(2- (2-(methylamino)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[001110] Mixture of tert-butyl N-[2-[2-[2-[[[(2S,4R)-4-hydroxy-1-[(2S)-3-methyl-2-(3-methylisoxazole-5 -yl)butanoyl]pyrrolidin-2-carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]ethoxy]ethyl]-N-methyl-carbamate (90 mg, 0.12 mmol, 1 equiv .) in hydrochloric acid/dioxane (4 M, 10 ml, 311 eq.) was degassed and purged with nitrogen 3 times, and then the mixture was stirred at 25°C for 0.5 hour under nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure to obtain a residue. Compound (2S,4R)-4-hydroxy-N-[[2-[2-[2-(methylamino)ethoxy]ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]-1-[ (2S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (75 mg, 0.11 mmol, 91% yield, hydrochloride) was obtained as a colorless oil. LC/MS (ESI) m/z: 600.4 [M+1] ⁺ .

[001111] Step 2: Preparation of tert-butyl 4-(6-chloro-8-fluoro-2-((3-((2-(2-(2-(((2S,4R)-4-hydroxy-1 -((S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)ethyl) methyl)amino)-3-oxopropyl)amino)-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazine-1-carboxylate

[001112] Mixture of (2S,4R)-4-hydroxy-N-[[2-[2-[2-(methylamino)ethoxy]ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]- 1-[(2S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (75 mg, 0.11 mmol, 1 eq., hydrochloride), 3-[[4 -(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]propanoic acid (70 mg, 0.11 mmol , 1 eq.), 1-hydroxybenzotriazole (23 mg, 0.17 mmol, 1.5 eq.), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (33 mg, 0.17 mmol, 1.5 eq. .) and N,N-diisopropylethylamine (61 mg, 0.47 mmol, 4 eq.) in N,N-dimethylformamide (1 ml) were degassed and purged with nitrogen 3 times, and then the mixture was stirred at 25°C for 12 hours in a nitrogen atmosphere. The reaction mixture was quenched by adding 10 ml of water and extracted with ethyl acetate (20 ml x 3). The combined organic layers were washed with brine (10 ml x 3), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (9% methanol in dichloromethane). tert-butyl 4-[6-chloro-8-fluoro-2-[[3-[2-[2-[2-[[[(2S,4R)-4-hydroxy-1-[(2R)-3 -methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]ethoxy]ethyl-methyl-amino]-3 -oxo-propyl]amino]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (75 mg, 0.06 mmol, 50% yield, 93% purity) was obtained as yellow oil. LC/MS (ESI) m/z: 590.4 [M/2+1] ⁺ .

[001113] Step 3: Preparation of (2S,4R)-N-(2-(2-(2-(3-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-4 -(piperazin-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((S) -3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide

[001114] Mixture of tert-butyl 4-[6-chloro-8-fluoro-2-[[3-[2-[2-[2-[[[(2S,4R)-4-hydroxy-1-[( 2S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]ethoxy]ethyl-methyl- amino]-3-oxo-propyl]amino]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (80 mg, 0.06 mmol, 1 eq.) and trifluoroacetic acid (821 mg, 7.20 mmol, 106.05 eq.) in dichloromethane (5 ml) was degassed and purged with nitrogen 3 times, and then the mixture was stirred at 25 °C for 1 hour under nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure to obtain a residue. Compound (2S,4R)-N-[[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1- yl-quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]-4-hydroxy-1-[(2S)-3- Methyl 2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (75 mg, 0.06 mmol, 92% yield, trifluoroacetate) was obtained as a yellow oil.

[001115] Step 4: Preparation of (2S,4R)-N-(2-(2-(2-(3-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7 -(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-(( S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide

[001116] To a solution of (2S,4R)-N-[[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4- piperazin-1-yl-quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]-4-hydroxy-1-[(2S )-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (75 mg, 0.06 mmol, 1 eq., trifluoroacetate) and 2,6-lutidine (67 mg, 0 .62 mmol, 73 μl, 10 eq.) in dichloromethane (20 ml) a solution of prop-2-enoyl chloride (6 mg, 0.06 mmol, 5.13 μl, 1 eq.) in dichloromethane (1 ml) was added dropwise at minus 78°C in a nitrogen atmosphere. The reaction solution was stirred at minus 78°C for 0.5 hours. The reaction mixture was quenched by adding 10 ml of water and extracted with ethyl acetate (20 ml x 3). The combined organic layers were washed with brine (10 ml x 3), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by semipreparative reverse phase HPLC. 2S,4R)-N-[[2-[2-[2-[3-[[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop-2 -enoylpiperazin-1-yl)quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]-4-hydroxy-1-[( 2S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (33.7 mg, 0.03 mmol, 47% yield, 99% purity) was obtained as a white solid . LC/MS (ESI) m/z: 566.3 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.01 (br s, 1H), 8.97 (s, 1H), 8.33 (br s, 1H), 7.79 (br d, J = 6.0 Hz, 2H), 7.46-7.38 (m, 1H), 7.34 (br d, J = 7.8 Hz, 1H), 7.29-7.17 (m, 4H), 7.11-6.95 (m, 3H), 6.84 (br dd, J = 10.5, 16.7 Hz, 1H), 6.24-6.14 (m, 2H), 5.74 (br d, J = 10.6 Hz, 1H), 5.08 (br s, 1H), 4.48-4.12 (m, 6H), 3.88-3.72 (m, 12H), 3.60-3.50 (m, 5H), 3.22-2.83 (m, 4H), 2.71-2.57 (m, 3H), 2.47-2.41 ( m, 3H), 2.30-2.27 (m, 1H), 2.25-2.19 (m, 2H), 2.15-2.09 (m, 1H), 2.07-1, 99 (m, 1H), 1.97-1.84 (m, 1H), 0.99-0.88 (m, 3H), 0.81-0.72 (m, 3H).

[001117] Example of the synthesis of (2S,4R)-1-((2S,E)-2-(tert-butyl)-17-(4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1 -yl)quinazolin-4-yl)piperazin-1-yl)-4,17-dioxo-6,9,12-trioxa-3-azaheptadec-15-enoyl)-4-hydroxy-N-((S)- 1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (representative compound 374)

[001118] Step 1: Preparation of 2-(2-(3-(benzyloxy)propoxy)ethoxy)ethan-1-ol

[001119] To a mixture of 2-[2-[2-(3-benzyloxypropoxy)ethoxy]ethoxy]tetrahydropyran (6.3 g, 18.61 mmol, 1 eq.) in methanol (70 ml) was added p-toluenesulfonic acid ( 641 mg, 3.72 mmol, 0.2 eq.). The mixture was stirred at 20°C for 12 hours. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by chromatography on silica gel (petroleum ether: ethyl acetate in a ratio of 4:1 to 1:1). The 2-[2-(3-benzyloxypropoxy)ethoxy]ethanol compound (3.9 g, 15.33 mmol, 82% yield) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.35-7.28 (m, 5H), 4.51 (s, 2H), 3.72-3.71 (m, 2H), 3.62 -3.60 (m, 2H), 3.59-3.56 (m, 6H), 1.94-1.86 (m, 2H).

[001120] Step 2: Preparation of ethyl 1-phenyl-2,6,9,12-tetraoxatetradecane-14-oate

[001121] To a mixture of 2-[2-(3-benzyloxypropoxy)ethoxy]ethanol (4.5 g, 17.69 mmol, 1 eq.) and rhodium acetate (391 mg, 0.88 mmol, 0.05 eq. ) in dichloromethane (50 ml) was added dropwise ethyl 2-diazoacetate (12.11 g, 106.16 mmol, 6 eq.) in dichloromethane (10 ml) at 0°C under nitrogen atmosphere. The mixture was stirred at 20°C for 12 hours under a nitrogen atmosphere. The mixture was quenched with acetic acid (1 ml). It was then washed with brine (40 ml×3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by chromatography on silica gel (petroleum ether: ethyl acetate in a ratio of 5:1 to 3:1). Ethyl 2-[2-[2-(3-benzyloxypropoxy)ethoxy]ethoxy]acetate (3.6 g, 10.58 mmol, 59% yield) was obtained as a light yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.34-7.27 (m, 5H), 4.51 (s, 2H), 4.22-4.20 (m, 2H), 4.14 (s, 2H), 3.78-3.70 (m, 4H), 3.59-3.58 (m, 2H), 3.57-3.56 (m, 6H), 1.93-1 .85 (m, 2H), 1.30-1.28 (m, 3H).

[001122] Step 3: Preparation of ethyl 2-(2-(2-(3-hydroxypropoxy)ethoxy)ethoxy)acetate

[001123] To a mixture of ethyl 2-[2-[2-(3-benzyloxypropoxy)ethoxy]ethoxy]acetate (3.6 g, 10.58 mmol, 1 eq.) in ethanol (40 ml) was added a palladium-activated catalyst carbon (0.4 g, purity 10%)) in a nitrogen atmosphere. The mixture was degassed and charged with hydrogen 3 times. The mixture was stirred at 50°C for 12 hours under a hydrogen atmosphere [0.35 MPa (50 psi)]. The mixture was filtered and concentrated under reduced pressure to obtain the product. Ethyl 2-[2-[2-(3-hydroxypropoxy)ethoxy]ethoxy]acetate (2.7 g) was obtained as a brown oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.19-4.11 (m, 2H), 3.74 (s, 2H), 3.72-3.69 (m, 4H), 3.65 -3.60 (m, 8H), 1.82-1.86 (m, 2H), 1.25 (t, J = 7.2 Hz, 3H).

[001124] Step 4: Preparation of ethyl 2-(2-(2-(3-oxopropoxy)ethoxy)ethoxy)acetate

[001125] To a mixture of ethyl 2-[2-[2-(3-hydroxypropoxy)ethoxy]ethoxy]acetate (2.7 g, 10.79 mmol, 1 eq.) in dichloromethane (30 ml) was added Dess-Martin Periodinan (6.86 g, 16.18 mmol, 5.01 ml, 1.5 eq.) at 0°C. The mixture was stirred at 20°C for 0.5 hour. The mixture was filtered. The filtrate was washed with saturated sodium dicarbonate solution (30 ml x 2) and brine (30 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated under pressure to obtain a residue. The residue was purified by chromatography on silica gel (petroleum ether: ethyl acetate in a ratio of 3:1 to 1:1). Ethyl 2-[2-[2-(3-oxopropoxy)ethoxy]ethoxy]acetate compound (2.5 g, 10.07 mmol, 93% yield) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) 5 9.77 (s, 1H), 4.20-4.18 (m, 2H), 4.12 (s, 2H), 3.82-3.79 (m, 2H), 3.71-3.69 (m, 4H), 3.67-3.61 (m, 4H), 2.66 (t, J = 6.0 Hz, 2H), 1, 25 (t, J = 3.6 Hz, 3H).

[001126] Step 5: Preparation of tert-butyl (E)-5-(2-(2-(2-ethoxy-2-oxoethoxy)ethoxy)ethoxy)pent-2-enoate

[001127] Mixture of ethyl 2-[2-[2-(3-oxopropoxy)ethoxy]ethoxy]acetate (2.5 g, 10.07 mmol, 1 eq.) and tert-butyl 2-(triphenylphosphanylidene) acetate (3.79 g, 10.07 mmol, 1 eq.) in dichloromethane (25 ml) was stirred at 20°C for 12 hours. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by chromatography on silica gel (petroleum ether: ethyl acetate in a ratio of 6:1 to 4:1). Tert-butyl (E)-5-[2-[2-(2-ethoxy-2-oxo-ethoxy)ethoxy]ethoxy]pent-2-enoate compound (2.48 g, 7.16 mmol, 71% yield ) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) 8 6.88-6.80 (m, 1H), 5.82-5.78 (m, 1H), 4.26-4.23 (m, 2H) , 4.21 (s, 2H), 3.73-3.71 (m, 4H), 3.64-3.60 (m, 2H), 3.59-3.56 (m, 4H), 2 .49-2.44 (m, 2H), 1.47 (s, 9H), 1.30-1.26 (m, 3H).

[001128] Step 6: Preparation of (E)-4-oxo-3,6,9,12-tetraoxaheptadec-15-ene-17-oic acid

[001129] To a mixture of tert-butyl (E)-5-[2-[2-(2-ethoxy-2-oxo-ethoxy)ethoxy]ethoxy]pent-2-enoate (1.0 g, 2.89 mmol , 1 eq.) in dichloromethane (15 ml) was added trifluoroacetic acid (3.00 ml). The mixture was stirred at 20°C for 1 hour. The mixture was concentrated under reduced pressure to obtain the product. (E)-5-[2-[2-(2-ethoxy-2-oxo-ethoxy)ethoxy]ethoxy]pent-2-enoic acid (1.4 g) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.18-7.11 (m, 1H), 5.96-5.92 (m, 1H), 4.25-4.23 (m, 2H) , 4.16 (s, 2H), 3.74-3.69 (m, 6H), 3.67-3.64 (m, 4H), 2.58-2.53 (m, 2H), 1 .29 (t, J = 7.2 Hz, 3H).

[001130] Step 7: Preparation of ethyl (E)-2-(2-(2-((5-(4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-4 -yl)piperazin-1-yl)-5-oxopent-3-en-1-yl)oxy)ethoxy)ethoxy)acetate

[001131] To a mixture of (E)-5-[2-[2-(2-ethoxy-2-oxo-ethoxy)ethoxy]ethoxy]pent-2-enoic acid (150 mg, 0.38 mmol, 1 eq. ) and 4-(6-chloro-8-fluoro-4-piperazin-1-yl-quinazolin-7-yl)naphthalene-2-ol (202 mg, 0.45 mmol, 1.17 eq., hydrochloride) in N,N-dimethylformamide (8 ml) was added 1-hydroxybenzotriazole (78 mg, 0.58 mmol, 1.5 eq.), N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (111 mg, 0.58 mmol, 1.5 eq.) and N,N-diisopropylethylamine (250 mg, 1.94 mmol, 0.33 ml, 5 eq.). The mixture was stirred at 20°C for 12 hours. The mixture was diluted with water (30 ml), extracted with ethyl acetate (25 ml × 3). The combined organic layer was washed with water (30 ml×2) and brine (30 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol = 10:1). Ethyl 2-[2-[2-[(E)-5-[4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazin-1-yl ]-5-oxo-pent-3-enoxy]ethoxy]ethoxy]acetate (120 mg, 0.17 mmol, 45% yield) was obtained as a yellow oil. LC/MS (ESI) m/z: 681.3 [M+1] ⁺ .

[001132] Step 8: Preparation of (E)-2-(2-(2-((5-(4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-4- yl)piperazin-1-yl)-5-oxopent-3-en-1-yl)oxy)ethoxy)ethoxy)acetic acid

[001133] To a mixture of ethyl 2-[2-[2-[(E)-5-[4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl] piperazin-1-yl]-5-oxo-pent-3-enoxy]ethoxy]ethoxy]acetate (120 mg, 0.17 mmol, 1 eq.) in tetrahydrofuran (5 ml) was added lithium hydroxide monohydrate (22 mg, 0 .52 mmol, 3 equiv.) in water (1 ml). The mixture was stirred at 0°C for 20 minutes. The mixture was diluted with water (10 ml), extracted with ethyl acetate (15 ml × 2). The aqueous phase was then adjusted to pH approximately 5 with hydrogen chloride solution (1 M). The mixture was extracted with ethyl acetate (10 ml x 3). The combined organic layer was washed with brine (20 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the product. Compound 2-[2-[2-[(E)-5-[4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazin-1-yl ]-5-oxo-pent-3-enoxy]ethoxy]ethoxy]acetic acid (110 mg) was obtained as a light yellow oil. LC/MS (ESI) m/z: 653.3 [M+1] ⁺ .

[001134] Step 9: Preparation of (2S,4R)-1-((2S,E)-2-(tert-butyl)-17-(4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene -1-yl)quinazolin-4-yl)piperazin-1-yl)-4,17-dioxo-6,9,12-trioxa-3-azaheptadec-15-enoyl)-4-hydroxy-N-((S )-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001135] To a mixture of 2-[2-[2-[(E)-5-[4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine -1-yl]-5-oxo-pent-3-enoxy]ethoxy]ethoxy]acetic acid (110 mg, 0.17 mmol, 1 eq.) and (2S,4R)-1-[(2S)-2 -amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (81 mg, 0.17 mmol, 1 eq., hydrochloride) in N,N-dimethylformamide (3 ml) added 1-hydroxybenzotriazole (34 mg, 0.25 mmol, 1.5 eq.), N-(3-dimethylaminopropyl)-N -ethylcarbodiimide hydrochloride (48 mg, 0.25 mmol, 1.5 eq.) and N,N-diisopropylethylamine (65 mg, 0.50 mmol, 0.09 ml, 3 eq.). The mixture was stirred at 20°C for 12 hours. The mixture was diluted with water (15 ml), extracted with ethyl acetate (15 ml × 3). The combined organic layer was washed with water (20 ml×2) and brine (20 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol = 10:1) to obtain a residue. The residue was purified by semipreparative reverse phase HPLC. (2S,4R)-1-[(2S)-2-[[2-[2-[2-[(E)-5-[4-[6-chloro-8-fluoro-7-(3-hydroxy -1-naphthyl)quinazolin-4-yl]piperazin-1-yl]-5-oxo-pent-3-enoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy- N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (50.8 mg, 0.04 mmol, 27% yield, 99% purity) was obtained as a white solid. LC/MS (ESI) m/z: 1195.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 8.84 (s, 1H), 8.66 (s, 1H), 8.08 (d, J = 1.2 Hz, 1H), 7.75 (d, J = 8.4 Hz, 1H), 7.42-7.38 (m, 5H), 7.27 (d, J = 2.4 Hz, 1H), 7.25-7.20 ( m, 2H), 7.06 (s, 1H), 6.90-6.86 (m, 1H), 6.58-6.54 (m, 1H), 5.02-4.98 (m, 1H), 4.56-4.54 (m, 1H), 4.42 (s, 1H), 4.05-4.03 (m, 6H), 3.89-3.71 (m, 5H) , 3.69-3.64 (m, 12H), 2.56-2.51 (m, 2H), 2.45 (s, 3H), 2.18-2.16 (m, 1H), 1 .99-1.95 (m, 1H), 1.50-1.46 (m, 3H), 1.04 (s, 9H).

[001136] Example of synthesis of (2S,4R)-1-((2S)-2-(3-(2-(2-(2-(3-((4-(4-acetylpiperazin-1-yl)-6 -chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)ethoxy)isoxazol-5-yl)-3-methylbutanoyl)-4 -Hydroxy-N-((S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (typical compound 386)

[001137] Step 1: Preparation of (2S,4R)-4-hydroxy-N-((S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)-1-( (S)-3-methyl-2-(3-(2-(2-(2-(methylamino)ethoxy)ethoxy)ethoxy)isoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide hydrochloride

[001138] To a solution of tert-butyl N-[2-[2-[2-[5-[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1- [2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2-methyl-propyl]isoxazol-3-yl]oxyethoxy]ethoxy]ethyl]-N- methyl carbamate (100 mg, 0.13 mmol, 1.0 eq) in dichloromethane (2 ml) added hydrochloric acid/dioxane (4 M, 2 ml, 61.91 eq). The reaction mixture was stirred at 20°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue. Desired compound (2S,4R)-4-hydroxy-N-[(1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-1-[(2S)-3 -methyl-2-[3-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]isoxazol-5-yl]butanoyl]pyrrolidine-2-carboxamide (90 mg, HCl) was obtained as a colorless oil .

[001139] Step 2: Preparation of (2S,4R)-1-((2S)-2-(3-(2-(2-(2-(3-((4-(4-acetylpiperazin-1-yl) -6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)ethoxy)isoxazol-5-yl)-3-methylbutanoyl) -4-hydroxy-N-((S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001140] To a solution of (2S,4R)-4-hydroxy-N-[(1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-1-[(2S )-3-methyl-2-[3-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]isoxazol-5-yl]butanoyl]pyrrolidine-2-carboxamide (90 mg, 0.13 mmol , 1.0 eq., HCl) in dimethylformamide (3 ml) was added diisopropylethylamine (50 mg, 0.4 mmol, 3.0 eq.). The mixture was stirred at 20°C for 10 minutes. Then 3-[[4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]propanoic acid (68 mg, 0.13 mmol, 1.0 eq.) and 1-hydroxybenzotriazole (34 mg, 0.25 mmol, 2.0 eq.) were added to the mixture and stirred at 20°C for 10 minutes. 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (48 mg, 0.25 mmol, 2.0 eq.) was added to the mixture and stirred at 20°C for 40 minutes. The mixture was diluted with water (2 ml), extracted with dichloromethane (20 ml) and then concentrated. The residue was purified by semipreparative reverse phase. (2S,4R)-1-[(2S)-2-[3-[2-[2-[2-[3-[[4-(4-acetylpiperazin-1-yl)-6-chloro-8- fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethoxy]isoxazol-5-yl]-3-methyl-butanoyl]-4-hydroxy -N-[(1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (26 mg, 0.021 mmol, 17% yield, 100% purity) was obtained as an off-white solid. LC/MS (EST) m/z: 1193.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 9.01-8.96 (m, 1H), 8.29 (d, J = 7.2 Hz, 1H), 7.81-7.73 (m, 2H), 7.41 (br s, 1H), 7.28-6.96 (m, 9H), 6.08 (d, J = 5.2 Hz, 1H), 5.12 (t , J = 7.2 Hz, 1H), 4.46-4.09 (m, 4H), 3.90-3.80 (m, 4H), 3.78-3.61 (m, 12H), 3.50 (d, J = 18.8 Hz, 10H), 3.04-2.96 (m, 2H), 2.80 (br s, 2H), 2.25 (d, J = 6.4 Hz, 2H), 2.10-1.96 (m, 5H), 1.78 (d, J = 6.8 Hz, 1H), 1.38 (d, J = 6.8 Hz, 1H), 1.26 (d, J = 6.8 Hz, 3H), 1.00-0.72 (m, 8H).

[001141] Example of the synthesis of (2S,4R)-1-((2R)-2-(3-(2-(2-(2-(3-((4-(4-acetylpiperazin-1-yl)-6 -chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)ethoxy)isoxazol-5-yl)-3-methylbutanoyl)-4 -Hydroxy-N-((S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (typical compound 387)

[001142] Step 1: Preparation of (2S,4R)-4-hydroxy-N-((S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)-1-( (R)-3-methyl-2-(3-(2-(2-(2-(methylamino)ethoxy)ethoxy)ethoxy)isoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide hydrochloride

[001143] To a solution of tert-butyl N-[2-[2-[2-[5-[(1R)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1- [2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2-methyl-propyl]isoxazol-3-yl]oxyethoxy]ethoxy]ethyl]-N- methyl carbamate (100 mg, 0.13 mmol, 1.0 eq.) in dichloromethane (2 ml) was added hydrochloric acid/dioxane (4 M, 2 ml, 61.91 eq.). The reaction mixture was stirred at 20°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue. Desired compound (2S,4R)-4-hydroxy-N-[(1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-1-[(2R)-3 -methyl-2-[3-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]isoxazol-5-yl]butanoyl]pyrrolidine-2-carboxamide (90 mg, HCl) was obtained as a colorless oil .

[001144] Step 2: Preparation of (2S,4R)-1-((2R)-2-(3-(2-(2-(2-(3-((4-(4-acetylpiperazin-1-yl) -6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)ethoxy)isoxazol-5-yl)-3-methylbutanoyl) -4-hydroxy-N-((S)-1-(2-methoxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001145] To a solution of (2S,4R)-4-hydroxy-N-[(1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-1-[(2R )-3-methyl-2-[3-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]isoxazol-5-yl]butanoyl]pyrrolidine-2-carboxamide (90 mg, 0.13 mmol , 1.0 eq., HCl) in dimethylformamide (3 ml) was added diisopropylethylamine (50 mg, 0.4 mmol, 3.0 eq.). The mixture was stirred at 20°C for 10 minutes. Then 3-[[4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]propanoic acid (71 mg, 0.13 mmol, 1.0 eq.) and 1-hydroxybenzotriazole (36.10 mg, 0.26 mmol, 2.0 eq.) were added to the mixture and stirred at 20°C for 10 minutes. 1-(3 β-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (51 mg, 0.26 mmol, 2.0 eq.) was added to the mixture and stirred at 20°C for 40 minutes. The mixture was diluted with water (2 ml), extracted with dichloromethane (20 ml), then concentrated. The residue was purified by semipreparative reverse phase. (2S,4R)-1-[(2R)-2-[3-[2-[2-[2-[3-[[4-(4-acetylpiperazin-1-yl)-6-chloro-8- fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethoxy]isoxazol-5-yl]-3-methyl-butanoyl]-4-hydroxy -N-[(1S)-1-[2-methoxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (45 mg, 0.035 mmol, 26% yield, 95% purity) was obtained as an off-white solid. LC/MS (ESI) m/z: 1193.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 9.01-8.96 (m, 1H), 8.29 (d, J = 7.2 Hz, 1H), 7.81-7.73 (m, 2H), 7.41 (br s, 1H), 7.28-6.96 (m, 9H), 6.08 (d, J = 5.2 Hz, 1H), 5.12 (t , J = 7.2 Hz, 1H), 4.46-4.09 (m, 4H), 3.90-3.80 (m, 4H), 3.78-3.61 (m, 12H), 3.50 (d, J = 18.8 Hz, 1H), 3.04-2.96 (m, 2H), 2.80 (br s, 2H), 2.25 (d, J = 6.4 Hz, 2H), 2.10-1.96 (m, 5H), 1.78 (d, J = 6.8 Hz, 1H), 1.38 (d, J = 6.8 Hz, 1H), 1.26 (d, J = 6.8 Hz, 3H), 1.00-0.72 (m, 8H).

[001146] Synthesis example of (2S,4R)-1-((2S)-21-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1 -yl)quinazolin-2-yl)amino)-2-(tert-butyl)-18-methyl-4-oxo-6,9,12,15-tetraoxa-3,18-diazagenicosanoyl)-4-hydroxy-N -((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (representative compound 396)

[001147] Step 1: Preparation of tert-butyl ((S)-20-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl) phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-4,21,21-trimethyl-18-oxo-7,10,13,16-tetraoxa-4,19-diazadocosyl)carbamate

[001148] (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]ethoxy ]acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (260 mg, 0.36 mmol , 1.00 eq., hydrochloride), tert-butylN-(3-bromopropyl) carbamate (170 mg, 0.71 mmol, 2.00 eq.) and diisopropylethylamine (93 mg, 0.71 mmol, 2.00 eq. .) placed in a test tube for reactions under the influence of microwave radiation in isopropanol (3 ml). The sealed tube was heated at 110°C for 2 hours under microwave radiation. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (silica gel plate, 13% dichloromethane in methanol) to give the compound tert-butylN-[3-[2-[2-[2-[2-[2-[[(1S)-1-[(2S ,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl ]amino]-2-oxo-ethoxy]ethoxy]ethoxy]ethoxy]ethyl-methyl-amino]propyl]carbamate (140 mg, 0.16 mmol, 46% yield) as a colorless oil. LC/MS (ESI) m/z: 849.5 [M+1] ⁺ .

[001149] Step 2: Preparation of (2S,4R)-1-((S)-21-amino-2-(tert-butyl)-18-methyl-4-oxo-6,9,12,15-tetraoxa- 3,18-diazagenicosanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001150] To a solution of tert-butyl N-[3-[2-[2-[2-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[ [(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy ]ethoxy]ethoxy]ethyl-methyl-amino]propyl]carbamate (140 mg, 0.16 mmol, 1.00 eq.) in dichloromethane (3 ml) was added hydrochloric acid in dioxane (4 M, 4 ml). The mixture was stirred at a temperature of 15°C for 0.5 hours. The reaction mixture was concentrated under reduced pressure to give the compound (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[3-aminopropyl(methyl)amino]ethoxy ]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] pyrrolidine-2-carboxamide (130 mg, hydrochloride) as a colorless oil. LC/MS (ESI) m/z: 749.4 [M+1] ⁺ .

[001151] Step 3: Preparation of tert-butyl 4-(7-bromo-6-chloro-8-fluoro-2-(((S)-20-((2S,4R)-4-hydroxy-2-(( (S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-4,21,21-trimethyl-18-oxo-7,10,13,16 -tetraoxa-4,19-diazadocosyl)amino)quinazolin-4-yl)piperazine-1-carboxylate

[001152] (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[3-aminopropyl(methyl)amino]ethoxy]ethoxy]ethoxy]ethoxy ]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide ( 130 mg, 0.16 mmol, 1.00 eq., hydrochloride), tert-butyl 4-(7-bromo-2,6-dichloro-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate (88 mg, 0.18 mmol, 1.15 eq.) and diisopropylethylamine (71 mg, 0.55 mmol, 0.10 mL, 3.46 eq.) were placed in a microwaving reaction tube in isopropanol (5 ml). The sealed tube was heated at 115°C for 3 hours under microwave radiation. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative thin layer chromatography (silica gel plate, 13% dichloromethane in methanol) to give tert-butyl4-[7-bromo-6-chloro-8-fluoro-2-[3-[2-[2-[2-[ 2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy]ethoxy]ethoxy]ethyl-methyl-amino]propylamino]quinazolin-4-yl]piperazin-1- carboxylate (120 mg, 0.1 mmol, 64% yield, 100% purity) as a colorless oil. LC/MS (ESI) m/z: 597.5 [M/2+1] ⁺ .

[001153] Step 4: Preparation of tert-butyl 4-(6-chloro-8-fluoro-2-(((S)-20-((2S,4R)-4-hydroxy-2-(((S)- 1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-4,21,21-trimethyl-18-oxo-7,10,13,16-tetraoxa-4 ,19-diazadocosyl)amino)-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazine-1-carboxylate

[001154] Mixture of tert-butyl 4-[7-bromo-6-chloro-8-fluoro-2-[3-[2-[2-[2-[2-[2-[[(1S)-1- [(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2- dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy]ethoxy]ethoxy]ethyl-methyl-amino]propylamino]quinazolin-4-yl]piperazine-1-carboxylate (120 mg, 0.10 mmol, 1.00 eq.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene-2-ol (54 mg, 0.20 mmol, 2.00 eq.), [2-(2-aminophenyl)phenyl]palladium(1+);dicyclohexyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane;methanesulfonate (9 mg, 0.01 mmol, 0.10 eq.) and potassium phosphate (1.5 M in H ₂ O, 0.20 ml, 3.00 eq.) in tetrahydrofuran (5 ml) were degassed and purged with nitrogen 3 times, and then the mixture was stirred at 60°C for 12 hours in a nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative thin layer chromatography (silica gel plate, 13% dichloromethane in methanol) to give tert-butyl 4-[6-chloro-8-fluoro-2-[3-[2-[2-[2-[2-[ 2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine -1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy]ethoxy]ethoxy]ethyl-methyl-amino]propylamino]-7-(3-hydroxy-1-naphthyl)quinazoline -4-yl]piperazine-1-carboxylate (100 mg, 0.08 mmol, 79% yield) as a yellow solid. LC/MS (ESI) m/z: 1255.8 [M+1] ⁺ .

[001155] Step 5: Preparation of (2S,4R)-1-((2S)-2-(tert-butyl)-21-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl )-4-(piperazin-1-yl)quinazolin-2-yl)amino)-18-methyl-4-oxo-6,9,12,15-tetraoxa-3,18-diazagenicosanoyl)-4-hydroxy-N -((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001156] To a solution of tert-butyl 4-[6-chloro-8-fluoro-2-[3-[2-[2-[2-[2-[2-[[(1S)-1-[(2S ,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl ]amino]-2-oxo-ethoxy]ethoxy]ethoxy]ethoxy]ethyl-methyl-amino]propylamino]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (100 mg , 0.08 mmol, 1.00 eq.) in dichloromethane (5 ml) was added hydrochloric acid in dioxane (4 M, 3 ml). The resulting mixture was stirred at a temperature of 20°C for 0.5 hours. The reaction mixture was concentrated under reduced pressure to give the compound (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[3-[[6-chloro-8 -fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]propyl-methyl-amino]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]- 3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (95 mg, hydrochloride) in as a yellow solid. LC/MS (EST) m/z: 1155.7 [M+1] ⁺ .

[001157] Step 6: Preparation of (2S,4R)-1-((2S)-21-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene -1-yl)quinazolin-2-yl)amino)-2-(tert-butyl)-18-methyl-4-oxo-6,9,12,15-tetraoxa-3,18-diazagenicosanoyl)-4-hydroxy -N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001158] Solution of (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[3-[[6-chloro-8-fluoro-7- (3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]propyl-methyl-amino]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl -butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (95 mg, 0.08 mmol, 1.00 eq., hydrochloride) and 2,6-lutidine (124 mg, 1.16 mmol, 0.14 ml, 15.00 eq.) in dichloromethane (5 ml) were cooled to minus 78°C. Prop-2-enoyl chloride (7.00 mg, 0.08 mmol, 1.00 eq) in dichloromethane (1 ml) was then added. The resulting mixture was stirred at a temperature of minus 78°C for 0.5 hours. The reaction mixture was quenched with saturated sodium bicarbonate solution (20 ml), then extracted with dichloromethane (20 ml x 2). The combined organic layers were washed with brine (20 ml×1), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by semipreparative reverse phase HPLC to give the compound (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[3-[[6-chloro- 8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]amino]propyl-methyl-amino]ethoxy]ethoxy]ethoxy ]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2- carboxamide (31 mg, 0.02 mmol, 30% yield, 98% purity), trifluoroacetic acid salt) as a yellow solid. LC/MS (ESI) m/z: 1209.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 9.58 (brs, 1H), 8.98 (s, 1H), 8.82-8.56 (m, 1H), 8.41 (d , J = 8.0 Hz, 1H), 8.08 (brs, 1H), 7.82 (d, J = 8.8 Hz, 1H), 7.59-7.18 (m, 9H), 7 .07 (s, 1H), 6.82 (dd, J = 11.2, 16.4 Hz, 1H), 6.20 (d, J = 16.4 Hz, 1H), 5.77 (d, J = 11.2 Hz, 1H), 4.96-4.85 (m, 2H), 4.54 (d, J = 9.6 Hz, 1H), 4.43 (t, J = 8.0 Hz, 1H), 4.28 (s, 1H), 4.24-4.07 (m, 3H), 3.97-3.66 (m, 9H), 3.63-3.09 (m, 20H), 2.81 (s, 3H), 2.45 (s, 3H), 2.21-1.91 (m, 3H), 1.90-1.70 (m, 1H), 1.50 -1.31 (m, 3H), 0.93 (s, 9H).

[001159] Synthesis example of (2S,4R)-1-((2S)-2-(2-(2-((1-((2R)-2-((4-(4-acryloylpiperazin-1-yl) -6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)acetamido)-3,3-dimethylbutanoyl)- 4-Hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (typical compound 398)

[001160] Step 1: Preparation of tert-butyl 4-(2-(2-ethoxy-2-oxoethoxy)ethoxy)piperidine-1-carboxylate

[001161] Diacetoxyrhodium (90 mg, 0.41 mmol, 0.05 eq.). Ethyl 2-diazoacetate (2.79 g, 24.46 mmol, 3 equiv) was added at 0°C and the mixture was stirred at 25°C for 12 hours. The reaction mixture was concentrated in vacuo. The crude product was purified by flash silica gel chromatography (petroleum ether:ethyl acetate 9:1 to 3:1). The compound tert-butyl 4-[2-(2-ethoxy-2-oxo-ethoxy)ethoxy]piperidine-1-carboxylate (1.7 g, 5.13 mmol, 63% yield) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.22 (q, J = 7.1 Hz, 2H), 4.16 (s, 2H), 3.83-3.71 (m, 4H), 3.70-3.63 (m, 2H), 3.50 (tt, J = 3.9, 8.2 Hz, 1H), 3.07 (ddd, J = 3.4, 9.5, 13 .3 Hz, 2H), 1.89-1.79 (m, 2H), 1.58-1.48 (m, 2H), 1.46 (s, 9H), 1.32-1.27 ( m, 3H).

[001162] Step 2: Preparation of 2-(2-((1-(tert-butoxycarbonyl)piperidin-4-yl)oxy)ethoxy)acetic acid

[001163] To a solution of tert-butyl 4-[2-(2-ethoxy-2-oxo-ethoxy)ethoxy]piperidine-1-carboxylate (1.6 g, 4.83 mmol, 1 eq.) in methanol (3 ml) and tetrahydrofuran (3 ml) and water (3 ml), lithium hydroxide monohydrate (405 mg, 9.66 mmol, 2 eq.) was added. The mixture was stirred at 25°C for 1 hour. 10 ml of water was added. The mixture was adjusted to pH 3-4 with 1M hydrochloric acid and then the aqueous phase was extracted with dichloromethane and methanol (10:1, 30 ml x 3). The combined organic phase was dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The compound 2-[2-[(1-tert-butoxycarbonyl-4-piperidyl)oxy]ethoxy]acetic acid (1.2 g, 3.96 mmol, 82% yield) was obtained as a yellow solid.

[001164] Step 3: Preparation of tert-butyl 4-(2-(2-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4- (4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)ethoxy)piperidine-1-carboxylate

[001165] To a solution of 2-[2-[(1-tert-butoxycarbonyl-4-piperidyl)oxy]ethoxy]acetic acid (410 mg, 1.35 mmol, 1 eq.) and (2S,4R)-1- [(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2 -carboxamide (600 mg, 1.35 mmol, 1 eq.) in N,N-dimethylformamide (10 ml) was added hydroxybenzotriazole (274 mg, 2.03 mmol, 1.50 eq.) and 1-(3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (389 mg, 2.03 mmol, 1.50 eq.) and N,N-diisopropylethylamine (593 mg, 4.59 mmol, 0.8 ml, 3.40 eq.). The mixture was stirred at 25°C for 12 hours. Water (50 ml) was added and the aqueous phase was extracted with ethyl acetate (40 ml x 3). The combined organic phase was washed with brine (20 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (dichloromethane:methanol = 1:0 to 20:1). Tert-butyl compound 4-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5 -yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy]piperidine-1-carboxylate (690 mg, 0.95 mmol, yield 70%) was obtained as a yellow oil. LC/MS (ESI) m/z: 730.4 [M+1] ⁺ .

[001166] Step 4: Preparation of (2S,4R)-1-((S)-3,3-dimethyl-2-(2-(2-(piperidin-4-yloxy)ethoxy)acetamido)butanoyl)-4- hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001167] A mixture of tert-butyl 4-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy]piperidine-1-carboxylate (690 mg, 0.95 mmol, 1.00 eq.) in hydrochloric acid/dioxane (4.0 M, 15 ml, 63.47 eq.) was stirred at 20°C for 1.0 hour. The solvent was removed under reduced pressure. The residue was diluted with methanol (10 ml) and acetonitrile (30 ml), the solvent was again removed and dried in vacuum. A suspension of (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-(4-piperidyloxy)ethoxy]acetyl]amino]butanoyl]-4-hydroxy-N-[ (1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (470 mg, 705.42 µmol, 1 eq., hydrochloride) and potassium carbonate (975 mg, 7 .05 mmol, 10.00 eq.) in a mixture of dichloromethane (8 ml) and acetonitrile (16 ml) was stirred at 25°C for 1.5 hours. The suspension was filtered through a celite pad and washed with dichloromethane (15 ml), the filtrate was concentrated and dried in vacuo. Compound (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-(4-piperidyloxy)ethoxy]acetyl]amino]butanoyl]-4-hydroxy-N-[ (1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (295 mg, 0.47 mmol, 66% yield) was obtained as a pale yellow solid.

[001168] Step 5: Preparation of tert-butyl 4-(6-chloro-8-fluoro-2-(((R)-1-(4-(2-(2-(((S)-1-(( 2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl- 1-oxobutan-2-yl)amino)-2-oxoethoxy)ethoxy)piperidin-1-yl)propan-2-yl)oxy)-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazine -1-carboxylate

[001169] To a solution of tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxo-ethoxy]quinazoline- 4-yl]piperazine-1-carboxylate (60 mg, 0.10 mmol, 1 eq.) and (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2 -(4-piperidyloxy)ethoxy]acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide ( 78 mg, 0.12 mmol, 1.2 eq.) in methanol (1 ml) and dichloromethane (2 ml), acetic acid (12 mg, 0.20 mmol, 2 eq.) was added, then sodium cyanoborohydride (19 mg, 0.31 mmol, 3 eq.) was added at 0°C. The mixture was stirred at 25°C for 3 hours. The mixture was concentrated in vacuo. The mixture was purified by preparative TLC (dichloromethane:methanol = 10:1) to obtain the product. Tert-butyl compound 4-[6-chloro-8-fluoro-2-[(1R)-2-[4-[2-[2-[[(1S)-1-[(2S,4R)-4- hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl]amino]-2- Oxo-ethoxy]ethoxy]-1-piperidyl]-1-methyl-ethoxy]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (63 mg) was obtained as a yellow solid substances. LC/MS (ESI) m/z: 1194.2 [M+1] ⁺ .

[001170] Step 6: Preparation of (2S,4R)-1-((2S)-2-(2-(2-((1-((2R)-2-((6-chloro-8-fluoro-7 -(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)acetamido)-3,3-dimethylbutanoyl)- 4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001171] To a solution of tert-butyl 4-[6-chloro-8-fluoro-2-[(1R)-2-[4-[2-[2-[[(1S)-1-[(2S,4R )-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino ]-2-oxo-ethoxy]ethoxy]-1-piperidyl]-1-methyl-ethoxy]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (61 mg, 0 .05 mmol, 1 eq.) in dichloromethane (2 ml) was added trifluoroacetic acid (770 mg, 6.75 mmol, 0.5 ml, 132.28 eq.). The mixture was stirred at 25°C for 0.5 hour. The mixture was concentrated in vacuo. Compound (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy- 1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N- [(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (61 mg, 0.05 mmol, 99% yield, trifluoroacetate) was obtained as a yellow oil.

[001172] Step 7: Preparation of (2S,4R)-1-((2S)-2-(2-(2-((1-((2R)-2-((4-(4-acryloylpiperazine-1- yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)acetamido)-3,3-dimethylbutanoyl )-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001173] To a solution of (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-( 3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethylbutanoyl]-4- Hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (61 mg, 0.05 mmol, 1 eq., trifluoroacetate) in dichloromethane ( 2 ml) 2,6-lutidine (54 mg, 0.50 mmol, 10 eq.) was added, then prop-2-enoyl chloride (4 mg, 0.045 mmol, 0.9 eq.) in dichloromethane (4 ml) was added at minus 65°C. The mixture was stirred at minus 65°C for 10 minutes. Water (10 ml) was added. The aqueous phase was extracted with dichloromethane (15 ml*3). The combined organic phase was concentrated in vacuo. The residue was purified by semipreparative reverse phase HPLC. The collected fraction was then concentrated to remove most of the acetonitrile. The solution was lyophilized. The compound (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy) was obtained -1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethylbutanoyl ]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (19 mg, 0.02 mmol, 31% yield, purity 99%, formate) as a white solid. LC/MS (ESI) m/z: 574.8 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.13-9.89 (m, 1H), 8.98 (s, 1H), 8.41 (br d, J = 7.1 Hz, 1H), 8.26 (s, 1H), 8.00 (s, 1H), 7.80 (d, J = 9.2 Hz, 1H), 7.42 (br d, J = 7.0 Hz , 3H), 7.39-7.26 (m, 4H), 7.21 (brd, J = 8.3 Hz, 2H), 7.06 (brd, J = 5.4 Hz, 1H), 6.83 (dd, J = 10.6, 16.9 Hz, 1H), 6.18 (br d, J = 16.9 Hz, 1H), 5.74 (br d, J = 10.6 Hz , 1H), 5.38 (br s, 1H), 5.12 (br s, 1H), 4.88 (br d, J = 6.1 Hz, 1H), 4.52 (d, J = 9 .9 Hz, 1H), 4.43 (t, J = 8.5 Hz, 1H), 4.27 (br s, 1H), 3.92 (br d, J = 8.4 Hz, 6H), 3.85 (br s, 2H), 3.78 (br s, 2H), 3.54 (br d, J = 14.1 Hz, 6H), 3.46-3.40 (m, 3H), 2.77 (br s, 2H), 2.61 (br s, 1H), 2.45 (s, 3H), 2.39 (br s, 1H), 2.15 (br s, 2H), 2 .02 (br d, J = 8.6 Hz, 1H), 1.77 (br s, 3H), 1.35 (br d, J = 6.5 Hz, 3H), 1.30 (br d, J = 6.1 Hz, 3H), 0.90 (br s, 9H).

[001174] Synthesis example of (2S,4R)-1-((2S)-2-(2-(2-((1-((2R)-2-((4-(4-acryloylpiperazin-1-yl) -6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)acetamido)-3,3-dimethylbutanoyl)- 4-Hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (representative compound 399)

[001175] Step 1: Preparation of tert-butyl 4-(2-(2-(2-ethoxy-2-oxoethoxy)ethoxy)ethoxy)piperidine-1-carboxylate

[001176] Diacetoxyrhodium (38 mg, 0.17 mmol, 0.05 eq.) in one portion at 0°C under nitrogen, then ethyl 2-diazoacetate (2.37 g, 20.73 mmol, 6 eq.) was added and stirred at 25°C for 16 hours. The mixture was quenched slowly with acetic acid (4 ml) and water (20 ml) was added. The aqueous phase was extracted with dichloromethane (15 ml x 3). The combined organic phase was washed with brine (20 ml x 3), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography (Si02, petroleum ether/ethyl acetate 30/1 to 10/1). The compound tert-butyl 4-[2-[2-(2-ethoxy-2-oxo-ethoxy)ethoxy]ethoxy]piperidine-1-carboxylate (700 mg, 1.86 mmol, 54% yield) was obtained as a colorless oil . ¹ H-NMR (400 MHz, CD ₃ OD) δ 4.22-4.16 (m, 4H), 3.71-3.67 (m, 10H), 3.60-3.56 (m, 1H ), 3.31-3.13 (m, 2H), 1.81-1.61 (m, 2H), 1.50-1.47 (m, 2H), 1.47 (s, 9H), 1.27 (t, J = 7.2 Hz, 3H).

[001177] Step 2: Preparation of 2-(2-(2-((1-(tert-butoxycarbonyl)piperidin-4-yl)oxy)ethoxy)ethoxy)acetic acid

[001178] To a solution of tert-butyl 4-[2-[2-(2-ethoxy-2-oxo-ethoxy)ethoxy]ethoxy]piperidine-1-carboxylate (700 mg, 1.86 mmol, 1 eq.) in Lithium hydroxide monohydrate (235 mg, 5.59 mmol, 3 eq.) was added to tetrahydrofuran (2 ml), methanol (2 ml) and water (2 ml), and the mixture was stirred at 25°C for 1 hour. Hydrochloric acid solution (1 M) was added to the mixture to adjust the pH to approximately 3~4. The reaction mixture was quenched with water (30 ml) and extracted with ethyl acetate (20 ml x 2), the combined organic layers were washed with brine (30 ml), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The compound 2-[2-[2-[(1-tert-butoxycarbonyl-4-piperidyl)oxy]ethoxy]ethoxy]acetic acid (460 mg, 1.32 mmol, 71% yield) was obtained as a colorless oil.

[001179] Step 3: Preparation of tert-butyl 4-(2-(2-(2-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1- (4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)ethoxy)ethoxy) piperidine-1-carboxylate

[001180] To a solution of (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (600 mg, 1.35 mmol, 1 eq.) in N,N-dimethylformamide (10 ml) was added 2-[2-[2-[(1 -tert-butoxycarbonyl-4-piperidyl)oxy]ethoxy]ethoxy]acetic acid (469 mg, 1.35 mmol, 1 eq.), 1-hydroxybenzotriazole (274 mg, 2.02 mmol, 1.5 eq.), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (388 mg, 2.02 mmol, 1.5 eq.) and N,N-diisopropylethylamine (593 mg, 4.59 mmol, 3.4 eq.), mixture stirred at 25°C for 16 hours. The reaction mixture was quenched with water (50 ml) and extracted with ethyl acetate (30 ml x 2), the combined organic layers were washed with brine (50 ml), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO ₂ , dichloromethane:methanol = 50:1 to 5:1). The residue was further purified by semipreparative reverse phase HPLC. Tert-butyl compound 4-[2-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (777 mg, 1 .00 mmol, yield 74%) was obtained as a colorless oil. LC/MS (ESI) m/z: 774.4 [M+1] ⁺ .

[001181] Step 4: Preparation of (2S,4R)-1-((S)-3,3-dimethyl-2-(2-(2-(2-(piperidin-4-yloxy)ethoxy)ethoxy)acetamido) butanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001182] To a solution of tert-butyl 4-[2-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4 -(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate ( 80 mg, 0.10 mmol, 1 eq.) in dichloromethane (2 ml) was added hydrochloric acid/dioxane (4 M, 0.8 ml, 30.96 eq.). The reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated in vacuo to give (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-[2-(4-piperidyloxy)ethoxy]ethoxy]acetyl]amino ]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (73 mg, 0.10 mmol, 99% yield , hydrochloride) as a light yellow solid.

[001183] Step 5: Preparation of tert-butyl 4-(6-chloro-8-fluoro-2-(((R)-1-(4-(2-(2-(2-(((S)-1 -((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3 -dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)ethoxy)ethoxy)piperidin-1-yl)propan-2-yl)oxy)-7-(3-hydroxynaphthalene-1-yl)quinazolin- 4-yl)piperazine-1-carboxylate

[001184] To a mixture of (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-[2-(4-piperidyloxy)ethoxy]ethoxy]acetyl]amino]butanoyl ]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (73 mg, 0.10 mmol, 1 eq., hydrochloric acid) in methanol (1 ml), sodium acetic acid (25 mg, 0.31 mmol, 3 eq.) was added. The reaction mixture was stirred at 25°C for 20 minutes. Solution of tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxo-ethoxy]quinazolin-4-yl] piperazine-1-carboxylate (60 mg, 0.10 mmol, 1 eq.) in dichloromethane (2 ml) was added, followed by acetic acid (12 mg, 0.21 mmol, 2 eq.). The reaction mixture was cooled to 0°C and sodium cyanoborohydride (6 mg, 0.41 mmol, 4 eq.) was added. The reaction mixture was stirred at 25°C for 14 hours. Dichloromethane (20 ml) and water (20 ml) were added and the mixture was separated. The organic layer was dried over sodium sulfate and then concentrated in vacuo to obtain a residue. The residue was purified by preparative TLC (10% methanol in dichloromethane) to give tert-butyl 4-[6-chloro-8-fluoro-2-[(1R)-2-[4-[2-[2-[2-[[ (1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl ]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy]ethoxy]-1-piperidyl]-1-methyl-ethoxy]-7-(3-hydroxy-1-naphthyl)quinazoline-4 -yl]piperazine-1-carboxylate (63 mg, 0.05 mmol, 46% yield, 94% purity) as a colorless gum. LC/MS (ESI) m/z. 1238.6 [M+1] ⁺ .

[001185] Step 6: Preparation of (2S,4R)-1-((2S)-2-(2-(2-(2-((1-((2R)-2-((6-chloro-8- fluoro-7-(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)ethoxy)acetamido)-3, 3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001186] To a solution of tert-butyl 4-[6-chloro-8-fluoro-2-[(1R)-2-[4-[2-[2-[2-[[(1S)-1-[( 2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-2-oxo-ethoxy]ethoxy]ethoxy]-1-piperidyl]-1-methyl-ethoxy]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (63 mg, 0.05 mmol, 1 eq.) in dichloromethane (5 ml) was added trifluoroacetic acid (7.70 g, 67.53 mmol, 5 ml, 1327.98 eq.). The reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated in vacuo to give (2S,4R)-1-[(2S)-2-[[2-[2-[2-[[1-[(2R)-2-[6-chloro-8- fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]ethoxy]acetyl]amino]-3,3- dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (63.7 mg, 0.05 mmol, 100% yield, trifluoroacetate), obtaining in the form of a light yellow gum.

[001187] Step 7: Preparation of (2S,4R)-1-((2S)-2-(2-(2-(2-((1-((2R)-2-((4-(4-acryloylpiperazine -1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)ethoxy)acetamido)- 3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001188] To a solution of (2S,4R)-1-[(2S)-2-[[2-[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro- 7-(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl- butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (63 mg, 0.05 mmol, 1 eq. trifluoroacetate) in N,N-dimethylformamide (1 ml) and dichloromethane (5 ml), 2,6-lutidine (184 mg, 1.72 mmol, 0.2 ml, 34.15 eq.) was added. The reaction mixture was cooled to minus 65°C, followed by a solution of prop-2-enoyl chloride (4 mg, 0.05 mmol, 0.9 eq.) in dichloromethane (0.36 ml). The reaction mixture was stirred at minus 65°C for 10 minutes. Dichloromethane (30 ml) and water (15 ml) were added and the mixture was separated. The organic layer was dried over sodium sulfate and then concentrated in vacuo to obtain a residue. This residue was purified by semipreparative reverse phase HPLC. The collected fractions were concentrated in vacuo to remove most of the acetonitrile and then lyophilized. (2S,4R)-1-[(2S)-2-[[2-[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3- hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]ethoxy]acetyl]amino]-3,3- dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (14.5 mg, 0.01 mmol, yield 21%, purity 98%, trifluoroacetate) was obtained as a white solid.

LC/MS (ESI) m/z: 1192.5 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.06 (br d, J=11.9 Hz, 1H), 9.26 (br s, 1H), 8.98 (s, 1H), 8.40 (d, J=7.7 Hz, 1H), 8.07 (s, 1H), 7.82 (d, J=8.2 Hz, 1H), 7.48-7.33 (m , 6H), 7.30 (d, J=2.2 Hz, 1H), 7.27-7.15 (m, 2H), 7.06 (dd, J=2.3, 9.0 Hz, 1H), 6.83 (dd, J=10.6, 16.6 Hz, 1H), 6.19 (dd, J=2.2, 16.6 Hz, 1H), 5.80-5.72 (m, 1H), 5.64 (br s, 1H), 4.89 (br s, 1H), 4.54 (br d, J=9.8 Hz, 1H), 4.42 (br t, J=8.0 Hz, 1H), 4.28 (br s, 1H), 3.96 (br d, J=7.3 Hz, 6H), 3.93-3.90 (m, 4H), 3.86 (br s, 5H), 3.81 (br s, 6H), 3.62-3.51 (m, 10H), 3.43-3.29 (m, 1H), 3.17 ( br s, 2H), 2.45 (s, 3H), 2.12-1.86 (m, 3H), 1.77 (br s, 1H), 1.47-1.33 (m, 6H) , 0.92 (s, 9H).

[001189] Synthesis example of (2S,4R)-1-((2S)-2-(2-(4-((1-((2R)-2-((4-(4-acryloylpiperazin-1-yl) -6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)methyl)piperazin-1-yl)acetamido)-3,3 -dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (representative compound 403)

[001190] Step 1: Preparation of tert-butyl 4-((4-(2-ethoxy-2-oxoethyl)piperazin-1-yl)methyl)piperidine-1-carboxylate

[001191] Sodium acetate (1.18 g, 14 .38 mmol, 3 eq.). The mixture was stirred at 25°C for 10 minutes. Then tert-butyl 4-formylpiperidine-1-carboxylate (1.02 g, 4.79 mmol, 1 eq.) and acetic acid (576 mg, 9.58 mmol, 0.6 mL, 2 eq.) were added with stirring at 25°C for 10 minutes. Sodium cyanoborohydride (603 mg, 9.58 mmol, 2 eq.) was then added at 0°C. The reaction mixture was then stirred at 25°C for 12 hours. The mixture was then diluted with water (30 ml). The mixture was then extracted with ethyl acetate (30 ml x 3). The combined organic layers were washed with water (50 ml x 2) and brine (50 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to leave a residue. The residue was purified by chromatography on silica gel (petroleum ether: ethyl acetate in the ratio from 7:3 to 0:1). The product tert-butyl 4-[[4-(2-ethoxy-2-oxo-ethyl)piperazin-1-yl]methyl]piperidine-1-carboxylate (1.7 g, 4.60 mmol, 96% yield) was obtained in the form of a colorless oil. LC/MS (ESI) m/z: 370.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.24-4.18 (m, 2H), 3.31 (s, 2H), 3.18-2.97 (m, 4H), 2.97 -2.90 (m, 4H), 2.77-2.67 (m, 4H), 2.09 (s, 2H), 1.94-1.85 (m, 1H), 1.84-1 .74 (m, 2H), 1.46 (s, 9H), 1.31-1.28 (m, 3H), 1.23-1.14 (m, 2H).

[001192] Step 2: Preparation of 2-(4-((1-(tert-butoxycarbonyl)piperidin-4-yl)methyl)piperazin-1-yl)acetic acid

[001193] To a mixture of tert-butyl 4-[[4-(2-ethoxy-2-oxo-ethyl)piperazin-1-yl]methyl]piperidine-1-carboxylate (200 mg, 0.54 mmol, 1 eq. ) in tetrahydrofuran (2 ml), methanol (1 ml) and water (2 ml) was added lithium hydroxide monohydrate (68 mg, 1.62 mmol, 3 eq.). The mixture was stirred at 25°C for 1 hour. The mixture was acidified with dilute hydrochloric acid (1 M) to pH=6-7. The mixture was then concentrated in vacuo to obtain a residue. The product 2-[4-[(1-tert-butoxycarbonyl-4-piperidyl)methyl]piperazin-1-yl]acetic acid (180 mg) was obtained as a pink solid.

[001194] Step 3: Preparation of tert-butyl 4-((4-(2-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4 -(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethyl)piperazin-1-yl) methyl)piperidine-1-carboxylate

[001195] To a mixture of 2-[4-[(1-tert-butoxycarbonyl-4-piperidyl)methyl]piperazin-1-yl]acetic acid (115 mg, 0.34 mmol, 1 eq.) and (2S,4R )-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl ]pyrrolidine-2-carboxamide (150 mg, 0.34 mmol, 1 eq., hydrochloride) in N,N-dimethylformamide (5 ml) was added 1-hydroxybenzotriazole (91 mg, 0.68 mmol, 2 eq.), N -(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (130 mg, 0.68 mmol, 2 eq.) and N,N-diisopropylethylamine (218 mg, 1.69 mmol, 0.3 ml, 5 eq.). The mixture was stirred at 25°C for 12 hours. The mixture was diluted with water (30 ml). The mixture was then extracted with ethyl acetate (30 ml x 3). The combined organic layers were washed with water (50 ml x 2) and brine (50 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to leave a residue. The residue was purified by preparative TLC (dichloromethane:methanol=10:1). Compound tert-butyl 4-[[4-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol- 5-yl)phenyl]ethyl]carbamoyl]pyrrolidin-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethyl]piperazin-1-yl]methyl]piperidine-1-carboxylate (70 mg ) was obtained as a colorless oil. LC/MS (ESI) m/z: 768.5 [M+1] ⁺ .

[001196] Step 4: Preparation of (2S,4R)-1-((S)-3,3-dimethyl-2-(2-(4-(piperidin-4-ylmethyl)piperazin-1-yl)acetamido)butanoyl )-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001197] To a mixture of tert-butyl 4-[[4-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-( 4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethyl]piperazin-1-yl]methyl]piperidin-1- carboxylate (70 mg, 0.09 mmol, 1 eq.) in dichloromethane (5 ml) was added hydrogen chloride/dioxane (4 M, 5 ml). The mixture was stirred at 25°C for 30 minutes. The mixture was then concentrated in vacuo to obtain a residue. Product (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[4-(4-piperidylmethyl)piperazin-1-yl]acetyl]amino]butanoyl]-4-hydroxy -N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (64 mg, hydrochloride) was obtained as a white solid. LC/MS (ESI) m/z: 668.5 [M+1] ⁺ .

[001198] Step 5: Preparation of tert-butyl 4-(6-chloro-8-fluoro-2-(((R)-1-(4-((4-(2-(((S)-1-( (2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl -1-oxobutan-2-yl)amino)-2-oxoethyl)piperazin-1-yl)methyl)piperidin-1-yl)propan-2-yl)oxy)-7-(3-hydroxynaphthalene-1-yl) quinazolin-4-yl)piperazine-1-carboxylate

[001199] To a mixture of (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[4-(4-piperidylmethyl)piperazin-1-yl]acetyl]amino]butanoyl] -4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (64 mg, 0.09 mmol, 1 eq., hydrochloride) sodium acetate (23 mg, 0.27 mmol, 3 eq.) was added to dichloromethane (3 ml) and methanol (3 ml). The mixture was stirred at 25°C for 10 minutes. Then tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[(1H)-1-methyl-2-oxo-ethoxy]quinazolin-4-yl] piperazine-1-carboxylate (53 mg, 0.09 mmol, 1 eq.) and acetic acid (11 mg, 0.18 mmol, 2 eq.) were added with stirring at 25°C for 10 minutes. Sodium cyanoborohydride (18 mg, 0.27 mmol, 3 eq.) was then added at 0°C. The reaction mixture was then stirred at 25°C for 12 hours. The mixture was concentrated in vacuo to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol=10:1). Product tert-butyl 4-[6-chloro-8-fluoro-2-[(1H)-2-[4-[[4-[2-[[(1S)-1-[(2S,4R)-4 -hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2 -oxo-ethyl]piperazin-1-yl]methyl]-1-piperidyl]-1-methyl-ethoxy]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazin-1-carboxylate (40 mg, 0.03 mmol, 36% yield) was obtained as a white solid. LC/MS (ESI)m/z: 617.4 [M/2+1] ⁺ .

[001200] Step 6: Preparation of (2S,4R)-1-((2S)-2-(2-(4-((1-((2R)-2-((6-chloro-8-fluoro-7 -(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)methyl)piperazin-1-yl)acetamido)-3,3 -dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001201] To a mixture of tert-butyl 4-[6-chloro-8-fluoro-2-[(1R)-2-[4-[[4-[2-[[(1S)-1-[(2S, 4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl] amino]-2-oxo-ethyl]piperazin-1-yl]methyl]-1-piperidyl]-1-methyl-ethoxy]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazin-1 -carboxylate (40 mg, 0.03 mmol, 1 eq.) in dichloromethane (5 ml) was added trifluoroacetic acid (3.08 g, 2 ml). The mixture was stirred at 25°C for 30 minutes. The mixture was concentrated in vacuo to obtain a residue. (2S,4R)-1-[(2S)-2-[[2-[4-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1 -naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxypropyl]-4-piperidyl]methyl]piperazin-1-yl]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy -N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (40 mg, trifluoroacetates) was obtained as a white solid.

[001202] Step 7: Preparation of (2S,4R)-1-((2S)-2-(2-(4-((1-((2R)-2-((4-(4-acryloylpiperazine-1- yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)methyl)piperazin-1-yl)acetamido)-3 ,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001203] To a mixture of (2S,4R)-1-[(2S)-2-[[2-[4-[[1-[(2R)-2-[6-chloro-8-fluoro-7-( 3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxypropyl]-4-piperidyl]methyl]piperazin-1-yl]acetyl]amino]-3,3-dimethylbutanoyl ]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (40 mg, 0.03 mmol, 1 eq., trifluoroacetates ) in dichloromethane (20 ml) was added 2,6-lutidine (35 mg, 0.32 mmol, 0.04 ml, 10 eq.). Prop-2-enoyl chloride (3 mg, 0.03 mL, 0.9 eq.) in dichloromethane (5 mL) was then added at minus 78° C. under nitrogen atmosphere. The mixture was then stirred at minus 78°C for 15 minutes under a nitrogen atmosphere. The mixture was diluted with water (25 ml). It was then extracted with dichloromethane (20 ml x 3). The combined organic layers were washed with brine (30 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to leave a residue. The residue was purified by preparative HPLC. Compound (2S,4R)-1-[(2S)-2-[[2-[4-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy- 1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]oxypropyl]-4-piperidyl]methyl]piperazin-1-yl]acetyl]amino]-3,3- dimethyl-butanoyl]-4-hydroxy-R-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (5.8 mg, 0.005 mmol, yield 14 .7%, 100% purity, formate) was obtained as a white solid. LC/MS (ESI) m/z: 593.8 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.96 (s, 1H), 8.44-8.42 (d, J=8.0 Hz, 1H), 8.28 (s, 2H ), 8.00 (s, 1H), 7.80-7.78 (d, J=8.0 Hz, 1H), 7.70-7.66 (d, J=16.0 Hz, 1H) , 7.46-7.39 (m, 3H), 7.38-7.31 (m, 2H), 7.28-7.27 (d, J=4.0 Hz, 1H), 7.26-7 .15 (m, 2H), 7.10-7.03 (m, 1H), 6.88-6.77 (m, 1H), 6.22-6.13 (m, 1H), 5.78 -5.71 (m, 1H), 5.45-5.36 (m, 1H), 4.92-4.81 (m, 1H), 4.50-4.32 (m, 3H), 4 .26 (s, 1H), 3.99-3.69 (m, 14H), 3.11-2.79 (m, 5H), 2.44 (s, 7H), 2.11-1.85 (m, 6H), 1.79-1.69 (m, 1H), 1.59-1.49 (m, 2H), 1.41-1.26 (m, 7H), 0.90 (s , 12H).

[001204] Example of the synthesis of (2S,4R)-1-((2S)-2-(2-(4-((4-((2R)-2-((4-(4-acryloylpiperazin-1-yl) -6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperazin-1-yl)methyl)piperidin-1-yl)acetamido)-3,3 -dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (representative compound 404)

[001205] Step 1: Preparation of tert-butyl 4-((1-((benzyloxy)carbonyl)piperidin-4-yl)methyl)piperazine-1-carboxylate

[001206] To a solution of tert-butyl 4-[2-(2-hydroxyethoxy)ethoxy]piperidine-1-carboxylate (1 g, 3.46 mmol, 1 eq.) in hydrochloride/methanol (4 M, 3 ml, 3.47 eq.), the mixture was stirred at a temperature of 25°C for 1 hour. The mixture was filtered and concentrated under reduced pressure to obtain a residue. The compound 2-[2-(4-piperidyloxy)ethoxy]ethanol (600 mg, hydrochloride) was obtained as a white solid.

[001207] Step 2: Preparation of tert-butyl 4-(piperidin-4-ylmethyl)piperazine-1-carboxylate

[001208] To a solution of tert-butyl 4-[(1-benzyloxycarbonyl-4-piperidyl)methyl]piperazine-1-carboxylate (910 mg, 2.18 mmol, 1 eq.) in methanol (15 ml) was added a palladium catalyst activated carbon (500 mg, purity 10%), the suspension was degassed in vacuum and purged with hydrogen several times. Palladium on activated carbon catalyst (100 mg, 0.07 mmol, 10% purity, 3.27e-2 eq.) in methanol (15 ml) was added to the mixture and stirred under a hydrogen atmosphere (4 mg, 2.18 mmol, 1 eq.) [0.35 MPa (50 psi)] at 25°C for 16 hours. The reaction mixture was filtered and the filter was concentrated. The compound tert-butyl 4-(4-piperidylmethyl)piperazine-1-carboxylate (1.8 g) was obtained as a colorless gum. LC/MS (ESI) m/z: 284.1 [M+1] ⁺ .

[001209] Step 3: Preparation of tert-butyl 4-((1-(2-ethoxy-2-oxoethyl)piperidin-4-yl)methyl)piperazine-1-carboxylate

[001210] To a solution of tert-butyl 4-(4-piperidylmethyl)piperazine-1-carboxylate (650 mg, 2.29 mmol, 1 eq.) and ethyl 2-bromoacetate (460 mg, 2.75 mmol, 0.3 ml, 1.2 eq.) in acetonitrile (10 ml) was added potassium carbonate (951 mg, 6.88 mmol, 3 eq.), the mixture was stirred at 80°C for 16 hours. The reaction mixture was quenched with water (30 ml) and extracted with ethyl acetate (20 ml x 2), the combined organic layers were washed with brine (30 ml), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by chromatography on silica gel (petroleum ether: ethyl acetate in a ratio of 10:1 to 0:1). The compound tert-butyl 4-[[1-(2-ethoxy-2-oxo-ethyl)-4-piperidyl]methyl]piperazine-1-carboxylate (537 mg, 1.45 mmol, 63% yield) was obtained as yellow oils ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.16 (q, J=7.2 Hz, 2H), 3.38 (d, J=4.8 Hz, 2H), 3.20 (s, 2H), 2.94 (d, J=11.6 Hz, 2H), 2.36-2.27 (m, 4H), 2.21-2.11 (m, 4H), 1.73 (d , J=12.8 Hz, 2H), 1.43 (s, 9H), 1.26-1.22 (m, 8H).

[001211] Step 4: Preparation of 2-(4-((4-(tert-butoxycarbonyl)piperazin-1-yl)methyl)piperidin-1-yl)acetic acid

[001212] To a solution of tert-butyl 4-[[1-(2-ethoxy-2-oxo-ethyl)-4-piperidyl]methyl]piperazine-1-carboxylate (567 mg, 1.53 mmol, 1 eq.) lithium hydroxide monohydrate (322 mg, 7.67 mmol, 5 eq.) was added to water (5 ml), methanol (5 ml) and tetrahydrofuran (5 ml) and the mixture was stirred at 25°C for 2 hours. The mixture was poured into hydrochloric acid solution (1 M) to adjust the pH to approximately 6~7. The reaction mixture was extracted (chloroform:isopropanol 5:1) (200 ml x 2), the combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The compound 2-[4-[(4-tert-butoxycarbonylpiperazin-1-yl)methyl]-1-piperidyl]acetic acid (332 mg, 0.97 mmol, 63% yield) was obtained as a yellow solid. ¹ H-NMR (400 MHz, CD ₃ OD) δ 3.57 (s, 2H), 3.42 (s, 4H), 2.38 (t, J=5.2 Hz, 4H), 2.26 (d, J=7.2 Hz, 2H), 2.02 (d, J=14.4 Hz, 2H), 1.56-1.42 (m, 11H), 1.15 (d, J= 6.0 Hz, 5H).

[001213] Step 5: Preparation of tert-butyl 4-((1-(2-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4 -(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethyl)piperidin-4-yl) methyl)piperazine-1-carboxylate

[001214] To a solution of 2-[4-[(4-tert-butoxycarbonylpiperazin-1-yl)methyl]-1-piperidyl]acetic acid (115 mg, 0.3 mmol, 1.08 eq.) in N,N -dimethylformamide (5 ml) was added (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-( 4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (150 mg, 0.3 mmol, 1 eq., hydrochloride), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (90 mg, 0 .47 mmol, 1.5 eq.), 1-hydroxybenzotriazole (63 mg, 0.47 mmol, 1.5 eq.), N,N-diisopropylethylamine (161 mg, 1.25 mmol, 0.2 ml, 4 eq.), the mixture was stirred at a temperature of 25°C for 2 hours. The reaction mixture was quenched with water (30 ml) and extracted with ethyl acetate (50 ml x 2), the combined organic layers were washed with brine (30 ml), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol=10:1). Compound tert-butyl 4-[[1-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol- 5 yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethyl]-4-piperidyl]methyl]piperazine-1-carboxylate (140 mg, 0 .18 mmol, yield 57%, purity 98%) was obtained as a colorless solid. LC/MS (ESI) m/z: 768.05 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 8.88 (s, 1H), 7.47-7.38 (m, 4H), 5.01 (q, J=7.2 Hz, 1H) , 4.65-4.44 (m, 4H), 3.90-3.83 (m, 1H), 3.80-3.69 (m, 1H), 3.42 (s, 3H), 3 .35 (s, 4H), 3.03 (d, J=4.4 Hz, 2H), 2.93-2.84 (m, 2H), 2.48 (s, 3H), 2.37 ( t, J=4.8 Hz, 4H), 2.25-2.20 (m, 4H), 2.02-1.91 (m, 1H), 1.80 (t, J=13.6 Hz , 2H), 1.60-1.50 (m, 4H), 1.45 (s, 9H), 1.33-1.22 (m, 2H), 1.06-1.02 (m, 9H ).

[001215] Step 6: Preparation of (2S,4R)-1-((S)-3,3-dimethyl-2-(2-(4-(piperazin-1-ylmethyl)piperidin-1-yl)acetamido)butanoyl )-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001216] To a solution of tert-butyl 4-[[1-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-( 4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethyl]-4-piperidyl]methyl]piperazine-1 -carboxylate (140 mg, 0.18 mmol, 1 eq.) in hydrochloric acid/methanol (4 M, 5 ml, 109.72 eq.), the mixture was stirred at 25°C for 1 hour. The mixture was concentrated under reduced pressure to obtain a residue. Compound (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[4-(piperazin-1-ylmethyl)-1-piperidyl]acetyl]amino]butanoyl]-4- Hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (120 mg, 0.17 mmol, 93% yield, hydrochloride) was obtained as colorless oil. LC/MS (ESI) m/z. 668.5 [M+1] ⁺ .

[001217] Step 7: Preparation of tert-butyl 4-(6-chloro-8-fluoro-2-(((R)-1-(4-((1-(2-(((S)-1-( (2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl -1-oxobutan-2-yl)amino)-2-oxoethyl)piperidin-4-yl)methyl)piperazin-1-yl)propan-2-yl)oxy)-7-(3-hydroxynaphthalene-1-yl) quinazolin-4-yl)piperazine-1-carboxylate

[001218] To a solution of (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[4-(piperazin-1-ylmethyl)-1-piperidyl]acetyl]amino]butanoyl ]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (73 mg, 0.1 mmol, 1 eq., hydrochloride ) in methanol (3 ml), sodium acetate (25 mg, 0.3 mmol, 3 eq.) was added and the mixture was stirred at 25°C for 0.5 h. Then tert-butyl 4-[6-chloro-8 -fluoro-7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxo-ethoxy]quinazolin-4-yl]piperazine-1-carboxylate (60 mg, 0 .1 mmol, 1 eq.) was added in dichloromethane (3 ml). Acetic acid (6 mg, 0.1 mmol, 0.006 ml, 1 eq.) followed by sodium cyanoborohydride (19 mg, 0.3 mmol, 3 eq.) was added to the reaction at 0°C for 0.5 hour. The mixture was stirred at 25°C for 11 hours. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol=10:1). Tert-butyl compound 4-[6-chloro-8-fluoro-2-[(1R)-2-[4-[[1-[2-[[(1S)-1-[(2S,4R)-4 -hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2 -oxo-ethyl]-4-piperidyl]methyl]piperazin-1-yl]-1-methyl-ethoxy]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazin-1-carboxylate (72 mg, 0.06 mmol, yield 55%, purity 97%) was obtained as a colorless oil. LC/MS (ESI) m/z: 617.4 [M/2+1] ⁺ .

[001219] Step 8: Preparation of (2S,4R)-1-((2S)-2-(2-(4-((4-((2R)-2-((6-chloro-8-fluoro-7 -(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl)oxy)propyl)piperazin-1-yl)methyl)piperidin-1-yl)acetamido)-3,3 -dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001220] To a solution of tert-butyl 4-[6-chloro-8-fluoro-2-[(1R)-2-[4-[[1-[2-[[(1S)-1-[(2S, 4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl] amino]-2-oxo-ethyl]-4-piperidyl]methyl]piperazin-1-yl]-1-methyl-ethoxy]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazin-1 -carboxylate (72 mg, 0.06 mmol, 1 eq.) in dichloromethane (10 ml), trifluoroacetic acid (1.54 g, 13.51 mmol, 1 ml, 231.28 eq.) was added and the mixture was stirred at 25 °C for 1 hour. The mixture was concentrated under reduced pressure to obtain a residue. Compound (2S,4R)-1-[(2S)-2-[[2-[4-[[4-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy- 1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxypropyl]piperazin-1-yl]methyl]-1-piperidyl]acetyl]amino]-3,3-dimethyl-butanoyl]-4- Hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (72 mg, 0.06 mmol, 99% yield, trifluoroacetate) was obtained as yellow oil. LC/MS (ESI) m/z: 561A [M/2+1] ⁺ .

[001221] Step 9: Preparation of (2S,4R)-1-((2S)-2-(2-(4-((4-((2R)-2-((4-(4-acryloylpiperazine-1- yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperazin-1-yl)methyl)piperidin-1-yl)acetamido)-3 ,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001222] To a solution of (2S,4R)-1-[(2S)-2-[[2-[4-[[4-[(2R)-2-[6-chloro-8-fluoro-7-( 3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxypropyl]piperazin-1-yl]methyl]-1-piperidyl]acetyl]amino]-3,3-dimethyl-butanoyl ]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (72 mg, 0.06 mmol, 1 eq., trifluoroacetate ) in N,N-dimethylformamide (1 ml), 2,6-lutidine (248 mg, 2.31 mmol, 0.3 ml, 40 eq.) was added, the mixture was cooled to minus 78°C, and then prop-2- enoyl chloride (4.70 mg, 0.05 mmol, 0.004 ml, 0.9 eq.) in dichloromethane (5 ml) was added dropwise and the mixture was stirred at minus 78°C for 30 minutes. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by semipreparative reverse phase HPLC. Compound (2S,4R)-1-[(2S)-2-[[2-[4-[[4-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy- 1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]oxypropyl]piperazin-1-yl]methyl]-1-piperidyl]acetyl]amino]-3,3- dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (9.9 mg, 0.008 mmol, yield 14 %, purity 96%) was obtained as a white solid. LC/MS (ESI) m/z: 594.8 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.98 (s, 1H), 8.45 (d, J=7.7 Hz, 1H), 8.25 (s, 2H), 8, 01 (s, 1H), 7.81 (d, J=8.2 Hz, 1H), 7.73 (d, J=9.7 Hz, 1H), 7.43 (d, J=8.2 Hz, 3H), 7.36 (d, J=8.4 Hz, 2H), 7.29 (d, J=2.2 Hz, 1H), 7.24-7.18 (m, 2H), 7.06 (dd, J=2.3, 8.5 Hz, 1H), 6.84 (dd, J=10.5, 16.7 Hz, 1H), 6.18 (dd, J=2, 1, 16.6 Hz, 1H), 5.79-5.72 (m, 1H), 5.45-5.38 (m, 1H), 4.88 (t, J=7.2 Hz, 1H ), 4.51-4.39 (m, 2H), 4.27 (s, 1H), 3.95-3.73 (m, 8H), 2.99 (d, J=14.9 Hz, 1H), 2.87-2.71 (m, 4H), 2.69-2.58 (m, 3H), 2.45 (s, 3H), 2.40 (s, 2H), 2.21 (s, 4H), 2.08-2.02 (m, 6H), 1.79-1.58 (m, 4H), 1.45-1.29 (m, 8H), 1.07 (s , 2H), 0.92 (s, 9H).

[001223] Example of the synthesis of (2S,4R)-1-((S)-20-((1-((R)-2-((4-(4-acryloylpiperazin-1-yl)-7-(3- hydroxynaphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)propyl)piperidin-4-yl)oxy)-2-(tert-butyl)- 4-oxo-6,9,12,15,18-pentaoxa-3-azaicozanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl) pyrrolidine-2-carboxamide (typical compound 410)

[001224] Step 1: Preparation of 2,4-dibromonaphthalene-1-amine

[001225] To a solution of dibromine (25.67 g, 160.63 mmol, 8.3 mL, 2.30 eq.) in acetic acid (75 mL) was added a solution of naphthalene-1-amine (10.00 g, 69.84 mmol, 9.8 ml, 1.00 eq.) in acetic acid (50 ml) at 5°C for 30 minutes. After addition was complete, acetic acid (50 ml) was diluted. The reaction mixture was heated to 70°C for 30 minutes. The suspension was filtered and washed with acetic acid (100 ml), the filter cake was suspended in 20% aqueous sodium hydroxide (120 ml), the mixture was stirred for 20 minutes and extracted with ethyl acetate (100 ml x 2), the combined organic phase was washed with sat. brine (150 ml), dried over sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 30/1 ~ 1/1). The elution solution was concentrated and treated with a mixture of ethyl acetate and petroleum ether (50 ml, V/V=l/10), the suspension was filtered and washed with petroleum ether (50 ml), and the filter cake was dried in vacuum. The compound 2,4-dibromonaphthalene-1-amine (10.20 g, 33.89 mmol, 49% yield) was obtained as a gray solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.18 (d, J=8.4 Hz, 1H), 7.81-7.90 (m, 2H), 7.62-7.53 (m , 2H), 4.63 (brs, 2H).

[001226] Step 2: Preparation of 4-bromo-2-hydroxynaphthalene-1-diazonium

[001227] To a solution of 2,4-dibromonaphthalene-1-amine (10.20 g, 33.89 mmol, 1.00 eq.) in a mixture of acetic acid (100 ml) and propionic acid (17 ml) was added sodium nitrate ( 2.69 g, 38.97 mmol, 1.15 eq.) at 5~8°C in portions for 15 minutes. The reaction mixture was stirred at 5~8°C for 45 minutes. The mixture was poured into ice water (660 ml) with stirring, the slurry was filtered and washed with water (100 ml), and the filtered precipitate was dried in air. The compound 4-bromo-2-hydroxynaphthalene-1-diazonium (8.20 g, 32.79 mmol, 97% yield) was obtained as a pale yellow solid.

[001228] Step 3: Preparation of 4-bromonaphthalene-2-ol

[001229] Sodium tetrahydroborate (2.84 g, 75.07 mmol, 2.29 eq.) in portions over 10 minutes at 10°C. The mixture was stirred at 20°C for 80 minutes. The solution was adjusted to pH~6 with hydrochloride solution (1.0 M, 80 ml), the organic phase was removed under reduced pressure and diluted with water (150 ml), the mixture was extracted with ethyl acetate (150 ml x 2), the combined organic phase was dried with sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 50/1 to 5/1). The compound 4-bromonaphthalene-2-ol (2.70 g, 12.10 mmol, 37% yield) was obtained as a purple solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.15 (d, J=8.0 Hz, 1H), 7.66 (d, J=7.6 Hz, 1H), 7.49-7, 41 (m, 3H), 7.15 (d, J=2.0 Hz, 1H), 5.42 (s, 1H).

[001230] Step 4: Preparation of 1-bromo-3-(methoxymethoxy)naphthalene

[001231] To a solution of 4-bromonaphthalene-2-ol (2.70 g, 12.10 mmol, 1.00 eq.) and potassium carbonate (5.02 g, 36.31 mmol, 3.00 eq.) in To N,N-dimethylformamide (20.0 mL), chloro(methoxy)methane (1.49 g, 18.51 mmol, 1.4 mL, 1.53 eq.) was added dropwise at 0°C. The mixture was stirred at 20°C for 12 hours. The mixture was diluted with water (100 ml) and extracted with ethyl acetate (50 ml x 2), the combined organic phase was washed with saturated brine (40 ml x 2), dried over sodium sulfate, filtered and concentrated. The crude product was purified by semipreparative reverse phase HPLC and the elution solution was removed under reduced pressure and dried in vacuo. The compound 1-bromo-3-(methoxymethoxy)naphthalene (1.40 g, 5.24 mmol, 43% yield) was obtained as a pale red oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.18-8.15 (m, 1H), 7.78-7.13 (m, 1H), 7.59 (d, J=2.4 Hz , 1H), 7.53-7.45 (m, 2H), 7.41 (d, J=2.0 Hz, 1H), 5.30 (s, 2H), 3.54 (s, 3H) .

[001232] Step 5: Preparation of 7-benzyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine-2,4-diol

[001233] Sodium (11.64 g, 506.31 mmol, 4.43 eq.) was added to ethanol (500 mL) in portions over 30 minutes at 0°C and stirred at 20°C for an additional 30 minutes. Then ethyl 1-benzyl-3-oxopiperidine-4-carboxylate (34.00 g, 114.18 mmol, 1.00 eq., hydrochloride salt) and urea (20.57 g, 342.54 mmol, 3.00 eq. .) was added to the above solution, the reaction mixture was stirred at 90°C for 12 hours. The mixture was adjusted to pH~7 with hydrochloride solution (1.0 M) and most of the organic phase was removed under reduced pressure. The suspension was filtered and washed with water (40 ml), and the filter cake was dried in vacuum. The compound 7-benzyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine-2,4-diol (18.10 g, 70.35 mmol, 62% yield) was obtained as a white solid. LC/MS (ESI) m/z: 258.1 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.93 (s, 1H), 10.61 (s, 1H), 7.37-7.27 (m, 5H), 3.62 (s , 2H), 3.13 (s, 2H), 2.61 (t, J=5.6 Hz, 2H), 2.23 (t, J=5.2 Hz, 2H).

[001234] Step 6: Preparation of 7-benzyl-2,4-dichloro-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine

[001235] To a solution of 7-benzyl-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2,4-diol (18.10 g, 70.35 mmol, 1.00 eq.) in phosphoryl trichloride (165.00 g, 1.08 mol, 100 mL, 15.30 eq.), diisopropylethylamine (27.28 g, 211.05 mmol, 36.8 mL, 3.00 eq.) was added slowly. The mixture was stirred at 110°C for 12 hours. The solvent was evaporated under reduced pressure and diluted with dichloromethane (150 ml), the suspension was slowly poured onto an ice bath and adjusted to pH ~ 7 with saturated sodium hydrogen carbonate solution (100 ml), the organic phase was separated and the aqueous phase was extracted with dichloromethane (150 ml × 3), the combined organic the phase was dried over sodium sulfate, filtered and concentrated (phosphoryl trichloride was slowly poured into water with stirring and adjusted to pH ~ 7 with a saturated solution of sodium bicarbonate). The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 10/1 to 5/1). The compound 7-benzyl-2,4-dichloro-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine (6.70 g, 22.78 mmol, 32% yield) was obtained as a pale red solid substances. Pyrimidine-2,4-diol (18.10 g, 70.35 mmol, 62% yield) was obtained as a white solid. LC/MS (ESI) m/z: 294.0 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.36-7.28 (m, 5H), 3.74 (s, 2H), 3.67 (s, 2H), 2.85 (s, 4H ).

[001236] Step 7: Preparation of tert-butyl 4-(7-benzyl-2-chloro-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazine-1-carboxylate

[001237] To a solution of 7-benzyl-2,4-dichloro-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine (28.50 g, 96.88 mmol, 1.00 eq.) and tert-butyl piperazine-1-carboxylate (18.95 g, 101.72 mmol, 1.05 eq.) in (methylsulfinyl)methane (200 ml) was added diisopropylethylamine (25.04 g, 193.76 mmol, 33.8 ml, 2.00 eq.). The mixture was stirred at 55°C for 3 hours. The mixture was diluted with water (500 ml) and ethyl acetate (60 ml), the suspension was stirred for 10 minutes and the aqueous phase was separated, the organic layer was filtered, the filter cake was washed with ethyl acetate (40 ml), petroleum ether (60 ml) and dried in vacuum, approximately 38 g of product was obtained as a white solid, the aqueous phase was extracted with ethyl acetate (150 ml x 2). The combined organic phase and the above filtrate were washed with water (300 ml), sat. brine (300 ml × 2), dried over sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate 10/1 to 3/1) to obtain approximately 3.2 g of product as a white solid. Compound tert-butyl 4-(7-benzyl-2-chloro-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl)piperazine-1-carboxylate (41.20 g, 92. 80 mmol, 96% yield) was obtained as a white solid. GS/MS (ESI) m/z: 444.1 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.36-7.29 (m, 5H), 3.70 (s, 2H), 3.62 (s, 2H), 3.54-3.49 (m, 8H), 2.68 (s, 4H), 1.50 (s, 9H).

[001238] Step 8: Preparation of tert-butyl (R)-4-(7-benzyl-2-((1,1-dimethoxypropan-2-yl)oxy)-5,6,7,8-tetrahydropyrido[3, 4-d]pyrimidin-4-yl)piperazine-1-carboxylate

[001239] To a solution of tert-butyl 4-(7-benzyl-2-chloro-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl)piperazine-1-carboxylate (10.00 g, 22.52 mmol, 1.00 eq) and (R)-1,1-dimethoxypropan-2-ol (4.06 g, 33.79 mmol, 1.50 eq) in toluene (120 ml) added tris(dibenzylideneacetone)dipalladium(0) (1.24 g, 1.35 mmol, 0.06 eq), (R)-(+)-2,2'-bis(diphenylphosphino)-1,1'- binaphthalene (1.40 g, 2.25 mmol, 0.10 eq.) and sodium 2-methylpropan-2-olate (5.41 g, 56.31 mmol, 2.50 eq.). The reaction mixture was degassed with nitrogen gas and stirred at 100°C for 4 hours. The solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography (17-50% ethyl acetate in petroleum ether). Compound tert-butyl 4-[7-benzyl-2-[(1R)-2,2-dimethoxy-1-methyl-ethoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4 -yl]piperazine-1-carboxylate (4.90 g, 9.29 mmol, 41% yield) was obtained as a green solid. LC/MS (ESI)w/z: 528.4 [M+1] ⁺ .

[001240] Step 9: Preparation of tert-butyl (R)-4-(2-((1,1-dimethoxypropan-2-yl)oxy)-5,6,7,8-tetrahydropyrido[3,4-d] pyrimidin-4-yl)piperazine-1-carboxylate

[001241] To a solution of tert-butyl 4-[7-benzyl-2-[(1R)-2,2-dimethoxy-1-methyl-ethoxy]-6,8-dihydro-5H-pyrido[3,4-d ]pyrimidin-4-yl]piperazine-1-carboxylate (4.90 g, 9.29 mmol, 1.00 eq.) to a mixture of methanol (90 ml) and tetrahydrofuran (15 ml) was added ammonia hydroxide (9.04 mmol , 92.86 mmol, 9.9 ml, 36% purity, 10.00 eq.) and palladium on activated carbon (10%), 1.00 g) under nitrogen gas. The suspension was degassed under vacuum and purged with hydrogen gas several times. The mixture was stirred under an atmosphere of hydrogen gas [0.35 MPa (50 psi)] at 50°C for 64 hours. The suspension was filtered through a celite pad and washed with methanol (30 ml), the filtrate was concentrated and dried in vacuum. Compound tert-butyl 4-[2-[(1R)-2,2-dimethoxy-1-methyl-ethoxy]-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl]piperazine -1-carboxylate (3.54 g, 8.09 mmol, 87% yield) was obtained as yellow gum. LC/MS (ESI) m/z: 438.2 [M+1] ⁺ .

[001242] Step 10: Preparation of tert-butyl (R)-4-(2-((1,1-dimethoxypropan-2-yl)oxy)-7-(3-(methoxymethoxy)naphthalene-1-yl)-5 ,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazine-1-carboxylate

[001243] To a mixture of tert-butyl 4-[2-[(1R)-2,2-dimethoxy-1-methyl-ethoxy]-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine-4 -yl]piperazine-1-carboxylate (3.54 g, 8.09 mmol, 1.00 eq) and 1-bromo-3-(methoxymethoxy)naphthalene (2.27 g, 8.50 mmol, 1.05 eq.) in dioxane (45 ml) was added RuPhos-Pd-G3 (338 mg, 0.40 mmol, 0.05 eq.), dicyclohexyl-(2',6'-diisopropoxy-[1,1'-biphenyl] -2-yl)phosphine (388 mg, 0.81 mmol, 0.10 eq.) and cesium carbonate (6.59 g, 20.23 mmol, 2.50 eq.). The reaction mixture was degassed with nitrogen gas and stirred at 100°C for 5 hours. The solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 5/1 ~ 3/1). Tert-butyl compound 4-[2-[(1R)-2,2-dimethoxy-1-methyl-ethoxy]-7-[3-(methoxymethoxy)-1-naphthyl]-6,8-dihydro-5H-pyrido [3,4-d]pyrimidin-4-yl]piperazin-1-carboxylate (3.90 g, 6.25 mmol, 77% yield) was obtained as a pale red solid. LC/MS (ESI) m/z: 624.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.11 (d, J=8.4 Hz, 1H), 7.76 (d, J=8.0 Hz, 1H), 7.46 (t, J=7.2 Hz, 1H), 7.38 (t, J=8.4 Hz, 1H), 7.17 (d, J=1.6 Hz, 1H), 6.89 (d, J= 2.0 Hz, 1H), 5.31 (s, 2H), 5.28-5.21 (m, 1H), 4.48 (d, J=6.0 Hz, 1H), 4.25 ( s, 2H), 3.59-3.43 (m, 17H), 3.37 (s, 2H), 2.88 (s, 2H), 1.51 (s, 9H), 1.37 (d , J=6.4 Hz, 3H).

[001244] Step 11: Preparation of (R)-2-((7-(3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)-5,6,7,8-tetrahydropyrido[3,4 -d]pyrimidin-2-yl)oxy)propanal

[001245] To a solution of tert-butyl 4-[2-[(1R)-2,2-dimethoxy-1-methyl-ethoxy]-7-[3-(methoxymethoxy)-1-naphthyl]-6,8-dihydro -5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (1.90 g, 3.05 mmol, 1.00 eq) in dichloromethane (10 ml) was added hydrochloride/dioxane ( 4.0 M, 20 ml, 26.26 eq.). The reaction mixture was stirred at 20°C for 4.0 hours. The suspension was filtered and washed with ethyl acetate (15 ml) and petroleum ether (15 ml), and the filter cake was dried in vacuum. Compound (2R)-2-[[7-(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl ]oxy]propanal (800 mg, 1.70 mmol, 56% yield, hydrochloride salt) was obtained as a pale yellow solid. LC/MS (ESI) m/z: 434.1 [M+1] ⁺ .

[001246] Step 12: Preparation of (R)-2-((4-(4-acryloylpiperazin-1-yl)-7-(3-hydroxynaphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3 ,4-d]pyrimidin-2-yl)oxy)propanal

[001247] To a solution of (2R)-2-[[7-(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine -2-yl]oxy]propanal (400 mg, 0.85 mmol, 1.00 eq. hydrochloride salt) was added to a mixture of dichloromethane (155 ml) and N,N-dimethylformamide (4 ml) with a solution of prop-2-enoyl chloride (85 mg, 0.94 mmol, 1.10 eq.) in dichloromethane (5 ml). And then 2,6-lutidine (2.74 g, 25.53 mmol, 30.00 eq.) was added dropwise at minus 78°C for 30 minutes. The reaction mixture was stirred at minus 78°C for 1.0 hour. The solvent was removed under reduced pressure, and then the high boiling solvent was evaporated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 3/1 to 1/1, ethyl acetate/methanol 20/1 to 10/1). Product (2R)-2-[[7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)-6,8-dihydro-5H-pyrido[3,4 -d]pyrimidin-2-yl]oxy]propanal (700 mg) was obtained as a pale red solid. LC/MS (ESI) m/z: 488.1 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 9.94 (s, 1H), 8.01 (d, J=8.8 Hz, 2H), 7.49-7.24 (m, 3H), 6.87-6.77 (m, 1H), 6.18 (d, J=16.8 Hz, 2H), 5.76 (d, J=6.4 Hz, 1H), 5.21-5 .05 (m, 1H), 4.60-4.46 (m, 2H), 4.05-3.95 (m, 4H), 3.83-3.74 (m, 8H), 1.50 (t, J=6.4 Hz, 3H).

[001248] Step 13: Preparation of 14-hydroxy-3,6,9,12-tetraoxatetradecyl 4-methylbenzenesulfonate

[001249] To a solution of 2-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]ethanol (15 g, 62.95 mmol, 13.27 ml, 2 eq.) in tetrahydrofuran (150 ml) sodium hydride (1.26 g, 31.48 mmol, 60% purity, 1 eq.) was added at 0°C. The mixture was stirred at 0°C for 0.5 hours. Then p-toluenesulfonyl chloride (6.00 g, 31.48 mmol, 1 eq.) was added and the mixture was stirred at 25°C for 2 hours. The mixture was poured into saturated aqueous ammonium chloride solution (100 ml), the aqueous layer was extracted with ethyl acetate (80 ml x 2). The organic phase was then dried over anhydrous sodium sulfate, filtered and the filtrate was condensed to obtain a residue. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 3/1 to dichloromethane/methanol 10/1) to give 2-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy ]ethoxy]ethyl 4-methylbenzenesulfonate (8.62 g, 21.96 mmol, 69% yield) as a light yellow oil.

[001250] Step 14: Preparation of 14-((tetrahydro-2H-pyran-2-yl)oxy)-3,6,9,12-tetraoxatetradecyl 4-methylbenzenesulfonate

[001251] To a solution of 2-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]ethyl 4-methylbenzenesulfonate (13 g, 33.12 mmol, 1 eq.) in dichloromethane (100 ml) added pyridine 4-methylbenzenesulfonate (416 mg, 1.66 mmol, 0.05 eq.) and 3,4-dihydro-2H-pyran (3.34 g, 39.75 mmol, 3.63 mL, 1.2 eq. .) at 0°C. The mixture was then stirred at 25°C for 16 hours. The mixture was filtered to obtain a filtrate. The filtrate was quenched with water (300 ml) and then diluted with dichloromethane (500 ml) and extracted with dichloromethane (500 ml x 2). The combined organic layers were washed with brine (300 ml), dried, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel chromatography (petroleum ether/ethyl acetate 3:1) to give 2-[2-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]ethoxy]ethyl 4-methylbenzenesulfonate (12.5 g, 26.23 mmol) as a yellow oil.

[001252] Step 15: Preparation of tert-butyl 4-((14-((tetrahydro-2H-pyran-2-yl)oxy)-3,6,9,12-tetraoxatetradecyl)oxy)piperidine-1-carboxylate

[001253] To a mixture of tert-butyl 4-hydroxypiperidine-1-carboxylate (6.33 g, 31.47 mmol, 1.2 eq.) in tetrahydrofuran (500 ml) was added sodium hydrogen (1.26 g, 31.47 mmol, purity 60%, 1.2 eq.) at 0°C in portions. The mixture was stirred at 0°C for 1 hour. A solution of 2-[2-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]ethoxy]ethyl 4-methylbenzenesulfonate (12.5 g, 26.23 mmol, 1 eq.) was then added to the mixture. in tetrahydrofuran (50 ml) drop by drop. The mixture was stirred at 25°C for 12 hours. The residue was poured into saturated sodium bicarbonate (300 ml). The mixture was extracted with ethyl acetate (100 ml x 3). The organic layer was combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated. The residue was purified by silica gel chromatography (petroleum ether/ethyl acetate 3:1). Tert-butyl 4-[2-[2-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate was obtained (13.3 g) as a yellow oil .

[001254] Step 16: Preparation of tert-butyl 4-((14-hydroxy-3,6,9,12-tetraoxatetradecyl)oxy)piperidine-1-carboxylate

[001255] To a solution of tert-butyl 4-[2-[2-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (13.3 g, 26.30 mmol, 1 eq.) in ethyl alcohol (50 ml) was added pyridine 4-methylbenzenesulfonate (661 mg, 2.63 mmol, 0.1 eq.). The mixture was stirred at 60°C for 12 hours. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (petroleum ether:ethyl acetate 5:1 to 0:1). Tert-butyl 4-[2-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (7.2 g, 17.08 mmol, 65% yield) obtained as a colorless oil.

[001256] Step 17: Preparation of tert-butyl 4-((17-oxo-3,6,9,12,15,18-hexaoxaicosyl)oxy)piperidine-1-carboxylate

[001257] To a solution of tert-butyl 4-[2-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (400 mg, 0.95 mmol, 1 eq.) in tetrahydrofuran (20 ml), sodium hydride (46 mg, 1.14 mmol, 60% purity, 1.2 eq.) was added in one portion at 0° C. under nitrogen atmosphere. The mixture was stirred at 0°C for 0.5 hours. Ethyl 2-bromoacetate (190 mg, 1.14 mmol, 1.2 eq.) was then added at 0°C. The mixture was stirred at 25°C for 11.5 hours. Water (30 ml) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 ml x 3). The combined organic phase was washed with brine (20 ml), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=5/1 to 1/1). Compound tert-butyl 4-[2-[2-[2-[2-[2-(2-ethoxy-2-oxo-ethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (250 mg , 0.49 mmol, 51% yield) was obtained as a white oil.

[001258] Step 18: Preparation of 17-((1-(tert-butoxycarbonyl)piperidin-4-yl)oxy)-3,6,9,12,15-pentaoxaheptadecanoic acid

[001259] To a solution of tert-butyl-4-[2-[2-[2-[2-[2-(2-ethoxy-2-oxo-ethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]piperidine-1 -carboxylate (250 mg, 492.50 µmol, 1 eq.) in tetrahydrofuran (5 ml) and water (5 ml) was added lithium hydrate (59 mg, 2.46 mmol, 5 eq.). The mixture was stirred at 25°C for 12 hours. The reaction mixture was charged with 1 M hydrochloric acid to adjust to pH=5 and extracted with 20 ml of ethyl acetate (20 ml * 3). The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure to give 2-[2-[2-[2-[2-[2-[(1-tert-butoxycarbonyl-4-piperidyl)oxy]ethoxy] ethoxy]ethoxy]ethoxy]ethoxy]acetic acid (200 mg) as a yellow oil.

[001260] Step 19: Preparation of tert-butyl 4-(((S)-19-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5 -yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-20,20-dimethyl-17-oxo-3,6,9,12,15-pentaoxa-18-azagenicosyl)oxy)piperidine-1-carboxylate

[001261] Mixture of 2-[2-[2-[2-[2-[2-[(1-tert-butoxycarbonyl-4-piperidyl)oxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]acetic acid (500 mg , 1.04 mmol, 1 eq.), (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1- [4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (464 mg, 1.04 mmol, 1 eq.), 1-hydroxybenzotriazole (211 mg, 1.56 mmol, 1.5 eq.), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (300 mg, 1.56 mmol, 1.5 eq.) and N,N-diisopropylethylamine (539 mg, 4.17 mmol, 4 eq.) in N,N-dimethylformamide (10 ml) was degassed and purged with nitrogen 3 times, and then the mixture was stirred at 25°C for 12 hours under N ₂ atmosphere. Water (50 ml) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 ml x 3). The combined organic phase was washed with brine (20 ml), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by semipreparative reverse phase HPLC to yield the compound tert-butyl 4-[2-[2-[2-[2-[2-[2-[[(1S)-1-[(2S,4R) -4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino] -2-oxoethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (700 mg, 0.77 mmol, 74% yield) as a colorless oil.

[001262] Step 20: Preparation of (2S,4R)-1-((S)-2-(tert-butyl)-4-oxo-20-(piperidin-4-yloxy)-6,9,12,15, 18-pentaoxa-3-azaicozanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001263] To a solution of tert-butyl 4-[2-[2-[2-[2-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[ [(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy ]ethoxy]ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (100 mg, 0.11 mmol, 1 eq.) in dichloromethane (2 ml) was added hydrochloric acid/dioxane (4 M, 1 ml, 36.25 eq. ). The reaction mixture was stirred at 25°C for 1 hour. The mixture was concentrated in vacuo to give (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-[2-[2-[2-[2-(4-piperidyloxy )ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2 -carboxamide (93 mg, hydrochloride) as a colorless gum.

[001264] Step 21: Preparation of (2S,4R)-1-((S)-20-((1-((R)-2-((4-(4-acryloylpiperazin-1-yl)-7-( 3-hydroxynaphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)propyl)piperidin-4-yl)oxy)-2-(tert-butyl) )-4-oxo-6,9,12,15,18-pentaoxa-3-azaicozanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl) ethyl)pyrrolidine-2-carboxamide

[001265] To a mixture of (R)-2-((4-(4-acryloylpiperazin-1-yl)-7-(3-hydroxynaphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3, 4-d]pyrimidin-2-yl)oxy)propanal (40 mg, 0.08 mmol, 1.00 eq.) and (2S,4R)-1-((S)-2-(tert-butyl)- 4-oxo-20-(piperidin-4-yloxy)-6,9,12,15,18-pentaoxa-3-azaicosan-1-oil)-4-hydroxy-N-((S)-1-(4 -(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (71 mg, 0.08 mmol, 1.00 eq., hydrochloride salt) in 1,2-dichloroethane (1.0 mL) and methanol (1.0 mL), sodium acetate (135 mg, 1.64 mmol, 20.00 eq.) was added over approximately 30 minutes. And then sodium cyanoborohydride (16 mg, 0.25 mmol, 3.00 eq.) was added at 0°C.

The mixture was allowed to warm to 20°C for 1.5 hours. The suspension was filtered and washed with dichloromethane (7 ml), the filtrate was concentrated and dried in vacuum. The residue was purified by preparative TLC (dichloromethane:methanol=15:1). The product was further purified by semipreparative reverse phase HPLC. The elution solution was lyophilized in vacuum. The product was further purified by semipreparative reverse phase HPLC. The elution solution was lyophilized in vacuum. Compound (2S,4R)-1-((S)-20-((1-((R)-2-((4-(4-acryloylpiperazin-1-yl)-7-(3-hydroxynaphthalene-1- yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)propyl)piperidin-4-yl)oxy)-2-(tert-butyl)-4-oxo- 6,9,12,15,18-pentaoxa-31-azaicosan-1-oil)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl) Pyrrolidine-2-carboxamide (7.1 mg, 0.005 mmol, 7% yield, 97% purity) was obtained as a pink solid. LC/MS (ESI) m/z: 1299.7 [M+23] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.60 (s, 1H), 7.94 (d, J=8.4 Hz, 1H), 7.55 (d, J=7.6 Hz, 1H), 7.48 (d, J=8.0 Hz, 1H), 7.34-7.21 (m, 7H), 6.82 (s, 1H), 6.71 (s, 1H), 6.51 (dd, J=10.8, 16.8 Hz, 1H), 6.26 (d, J=16.8 Hz, 1H), 5.68 (d, J=10.8 Hz, 1H ), 5.35-5.22 (m, 1H), 5.04-4.98 (m, 1H), 4.68 (t, J=8.0 Hz, 1H), 4.53 (d, J=8.8 Hz, 1H), 4.44 (s, 1H), 4.10 (s, 2H), 4.01-3.89 (m, 3H), 3.75-3.35 (m , 30H), 3.30-3.15 (m, 2H), 2.88-2.78 (m, 2H), 2.75-2.65 (m, 2H), 2.44 (s, 3H ), 2.41-2.33 (m, 2H), 2.27-2.15 (m, 2H), 2.08-2.00 (m, 1H), 1.96-1.86 (m , 3H), 1.56-1.44 (m, 3H), 1.40 (d, J=6.8 Hz, 3H), 1.26 (d, J=6.0 Hz, 3H), 0 .99 (s, 9H).

[001266] Example of the synthesis of 5-(2-(2-(2-((1-((2R)-2-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7 -(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindolin- 1,3-Dione (typical compound 428)

[001267] Step 1: Preparation of tert-butyl 4-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy )ethoxy)ethoxy)piperidine-1-carboxylate

[001268] To a mixture of tert-butyl 4-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (800 mg, 1.64 mmol, 1 eq.) and 2 -(2,6-dioxo-3-piperidyl)-5-hydroxy-isoindoline-1,3-dione (449 mg, 1.64 mmol, 1 eq.) in N,N-dimethylformamide (10 ml) added potassium carbonate (566 mg, 4.10 mmol, 2.5 eq.) and potassium iodide (27 mg, 0.16 mmol, 0.1 eq.). The mixture was stirred at 50°C for 12 hours. The mixture was poured into a solution of hydrochloric acid (30 ml, 1 M). It was then extracted with ethyl acetate (30 ml x 3). The combined organic layer was washed with water (50 ml×2), and brine (50 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate 3:1 to 1:2) to give the product. The product was further purified by preparative HPLC. The compound tert-butyl 4-[2-[2-[2-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]oxyethoxy]ethoxy]ethoxy]piperidin- 1-Carboxylate (330 mg, 0.56 mmol, 34% yield) was obtained as a white solid. LC/MS (ESI) m/z: 490.1 [M-100] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 7.81 (d, J=8.4 Hz, 1H), 7.44 (d, J=2.0 Hz, 1H), 7.36-7 .33 (m, 1H), 5.13-5.11 (m, 1H), 4.58 (s, 1H), 4.32-4.30 (m, 1H), 3.72-3.70 (m, 3H), 3.68-3.65 (m, 2H), 3.62-3.58 (s, 4H), 3.56-3.52 (m, 1H), 3.14-3 .02 (m, 2H), 2.78-2.77 (m, 1H), 2.74-2.73 (m, 2H), 2.06-2.12 (m, 1H), 1.84 -1.80 (m, 2H), 1.47-1.46 (m, 2H), 1.44 (s, 9H).

[001269] Step 2: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(2-(2-(2-(piperidin-4-yloxy)ethoxy)ethoxy)ethoxy)isoindoline-1,3 -dione

[001270] To a mixture of tert-butyl 4-[2-[2-[2-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]oxyethoxy]ethoxy] ethoxy]piperidine-1-carboxylate (330 mg, 0.55 mmol, 1 eq.) in dichloromethane (5 ml) was added hydrochloric acid/dioxane (4 M, 5 ml). The mixture was stirred at 25°C for 1 hour. The mixture was concentrated in vacuo to give the product 2-(2,6-dioxo-3-piperidyl)-5-[2-[2-[2-(4-piperidyloxy)ethoxy]ethoxy]ethoxy]isoindoline-1,3-dione (290 mg, 0.55 mmol, 98% yield, hydrochloride) as a yellow oil. LC/MS (ESI) m/z: 490.3 [M+1] ⁺ .

[001271] Step 3: Preparation of tert-butyl 4-(6-chloro-2-(((2R)-1-(4-(2-(2-(2-((2-(2,6-dioxopiperidin- 3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)piperidin-1-yl)propan-2-yl)oxy)-8-fluoro-7-(3-hydroxynaphthalene-1 -yl)quinazolin-4-yl)piperazine-1-carboxylate

[001272] To a mixture of 2-(2,6-dioxo-3-piperidyl)-5-[2-[2-[2-(4-piperidyloxy)ethoxy]ethoxy]ethoxy]isoindoline-1,3-dione (54 mg, 0.10 mmol, 1 eq., hydrochloride) in dichloromethane (3 ml) and methanol (3 ml) was added sodium acetate (25 mg, 0.30 mmol, 3 eq.). The mixture was stirred at 25°C for 10 minutes. Then tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxo-ethoxy]quinazolin-4-yl] piperazine-1-carboxylate (60 mg, 0.10 mmol, 1 eq.) and acetic acid (12 mg, 0.20 mmol, 0.01 ml, 2 eq.) were added with stirring at 25°C for 10 minutes . Sodium cyanoborohydride (19 mg, 0.30 mmol, 3 eq.) was then added at 0°C. The reaction mixture was then stirred at 25°C for 12 hours. The mixture was concentrated in vacuo to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol=10:1). The compound tert-butyl4-[6-chloro-2-[(1R)-2-[4-[2-[2-[2-[2-(2,6-dioxo-3-piperidyl)-1,3- dioxo-isoindolin-5-yl]oxyethoxy]ethoxy]ethoxy]-1-piperidyl]-1-methyl-ethoxy]-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazin- 1-Carboxylate (60 mg, 0.056 mmol, 55.10% yield) was obtained as a colorless oil. LC/MS (ESI) m/z: 1054.6 [M+1] ⁺ .

[001273] Step 4: Preparation of 5-(2-(2-(2-((1-((2R)-2-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl) -4-(piperazin-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindolin- 1,3-dione

[001274] To a mixture of tert-butyl 4-[6-chloro-2-[(1R)-2-[4-[2-[2-[2-[2-(2,6-dioxo-3-piperidyl) -1,3-dioxo-isoindolin-5-yl]oxyethoxy]ethoxy]ethoxy]-1-piperidyl]-1-methyl-ethoxy]-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-4 -yl]piperazine-1-carboxylate (60 mg, 0.056 mmol, 1 eq.) in dichloromethane (5 ml) was added trifluoroacetic acid (3.08 g, 27.01 mmol, 2 ml, 474 eq.). The mixture was stirred at 25°C for 30 minutes. The mixture was then concentrated to give the product 5-[2-[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4 -piperazin-1-yl-quinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]ethoxy]ethoxy]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione ( 60 mg, trifluoroacetate) as a colorless oil. LC/MS (ESI) m/z: 954.4 [M+1] ⁺ .

[001275] Step 5: Preparation of 5-(2-(2-(2-((1-((2R)-2-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro -7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl) isoindoline-1,3-dione

[001276] To a mixture of 5-[2-[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4- piperazin-1-yl-quinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]ethoxy]ethoxy]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione (60 mg, 0.056 mmol, 1 eq., trifluoroacetate) in dichloromethane (20 ml) was added 2,6-lutidine (0.06 ml, 10 eq.). Prop-2-enoyl chloride (4 mg, 0.050 mmol, 0.004 mL, 0.9 eq.) in dichloromethane (5 mL) was then added to the mixture at minus 78° C. under a nitrogen atmosphere. The mixture was stirred at minus 78°C for 30 minutes under a nitrogen atmosphere. The mixture was then quenched with water (20 ml). It was extracted with dichloromethane (20 ml x 3) and washed with brine (20 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC. Compound 5-[2-[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop -2-enoylpiperazin-1-yl)quinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]ethoxy]ethoxy]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3- dione (5.6 mg, 0.0052 mmol, 9.27% yield, 98% purity, formate) was obtained as a white solid. LC/MS (ESI) m/z: 504.8 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.10 (s, 1H), 8.31 (s, 2H), 7.99 (s, 1H), 7.80-7.78 (d , J=8.0 Hz, 2H), 7.47-7.40 (m, 2H), 7.36-7.30 (m, 1H), 7.28 (s, 1H), 7.23- 7.20 (m, 1H), 7.08-7.04 (m, 1H), 6.88-6.77 (m, 1H), 6.25-6.13 (m, 1H), 5, 83-5.67 (m, 2H), 5.43-5.31 (m, 1H), 5.13-5.06 (m, 1H), 4.29-4.24 (m, 2H), 3.94-3.72 (m, 12H), 3.58-3.52 (m, 7H), 3.24-3.13 (m, 2H), 2.94-2.71 (m, 3H ), 2.64-2.55 (m, 2H), 2.41-2.34 (m, 1H), 2.21-1.91 (m, 4H), 1.78-1.62 (m , 2H), 1.30-1.27 (m, 3H), 1.24-1.17 (m, 1H).

[001277] Example of the synthesis of 5-(2-(2-((2(2R)-2-((6-chloro-4-(4-(2,2-dihydroxyacetyl)piperazin-1-yl)-8-fluoro -7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindolin- 1,3-Dione (typical compound 434)

[001278] Step 1: Preparation of tert-butyl 4-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy) piperidine-1-carboxylate

[001279] To a solution of tert-butyl 4-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]piperidine-1-carboxylate (500 mg, 1.13 mmol, 1 eq.) in N,N-dimethylformamide ( 10 ml) 2-(2,6-dioxo-3-piperidyl)-5-hydroxy-isoindoline-1,3-dione (309 mg, 1.13 mmol, 1 eq.), potassium iodide (19 mg, 0 .11 mmol, 0.1 eq.) and potassium iodide (390 mg, 2.82 mmol, 2.5 eq.), the mixture was stirred at 50°C for 16 hours. The mixture was poured into hydrochloric acid solution (1 M) to adjust the pH to approximately 3~4, and the reaction mixture was extracted with ethyl acetate (50 ml). The combined organic layers were washed with brine (30 ml x 2), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol 10:1). The residue was purified by semipreparative reverse phase HPLC. Tert-butyl 4-[2-[2-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]oxyethoxy]ethoxy]piperidine-1-carboxylate (234 mg , 0.42 mmol, 38% yield) was obtained as a white solid. LC/MS (ESI) m/z: 568.3 [M+23] ⁺ .

[001280] Step 2: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(2-(2-(piperidin-4-yloxy)ethoxy)ethoxy)isoindoline-1,3-dione

[001281] To a solution of tert-butyl 4-[2-[2-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]oxyethoxy]ethoxy]piperidine-1 -carboxylate (330 mg, 0.60 mmol, 1 eq.) in dichloromethane (4 ml), hydrochloric acid/dioxane (4 M, 4 ml, 26.45 eq.) was added and the mixture was stirred at 25°C for 2 hours. The mixture was concentrated under reduced pressure to obtain a residue. Compound 2-(2,6-dioxo-3-piperidyl)-5-[2-[2-(4-piperidyloxy)ethoxy]ethoxy]isoindoline-1,3-dione (260 mg, 0.54 mmol, yield 89 %, hydrochloride) was obtained as a colorless oil. LC/MS (ESI) m/z: 446.1 [M+1] ⁺ .

[001282] Step 3: Preparation of tert-butyl 4-(6-chloro-2-(((2R)-1-(4-(2-(2-((2-(2,6-dioxopiperidin-3-yl )-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)piperidin-1-yl)propan-2-yl)oxy)-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin- 4-yl)piperazine-1-carboxylate

[001283] To a solution of 2-(2,6-dioxo-3-piperidyl)-5-[2-[2-(4-piperidyloxy)ethoxy]ethoxy]isoindoline-1,3-dione (86 mg, 0.19 mmol, 1.4 eq., hydrochloride) in methanol (3 ml) was added sodium acetate (34 mg, 0.41 mmol, 3 eq.), the mixture was stirred at 25°C for 0.5 h. Then tert -butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxo-ethoxy]quinazolin-4-yl]piperazin- 1-carboxylate (80 mg, 0.14 mmol, 1 eq.) in dichloromethane (3 ml). Acetic acid (8 mg, 0.14 mmol, 1 eq.) and sodium cyanoborohydride (26 mg, 0.41 mmol, 3 eq.) were added to the reaction at 0°C for 0.5 hour, the mixture was stirred at 25 °C for 11 hours. The reaction was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol 10:1). The compound tert-butyl 4-[6-chloro-2-[(1R)-2-[4-[2-[2-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo- isoindolin-5-yl]oxyethoxy]ethoxy]-1-piperidyl]-1-methyl-ethoxy]-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate ( 43 mg, 0.04 mmol, yield 28%, purity 90%) was obtained as a colorless oil. LC/MS (ESI)m/z: 1011.9 [M+1] ⁺ .

[001284] Step 4: Preparation of 5-(2-(2-((1-((2R)-2-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-4- (piperazin-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione

[001285] To a solution of tert-butyl 4-[6-chloro-2-[(1R)-2-[4-[2-[2-[2-(2,6-dioxo-3-piperidyl)-1, 3-dioxo-isoindolin-5-yl]oxyethoxy]ethoxy]-1-piperidyl]-1-methyl-ethoxy]-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazin- 1-carboxylate (43 mg, 0.04 mmol, 1 eq.) in dichloromethane (3 ml), hydrochloric acid/dioxane (4 M, 3 ml, 282 eq.) was added and the mixture was stirred at 25°C for 1 hour . The mixture was concentrated under reduced pressure to obtain a residue. Compound 5-[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazoline -2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]ethoxy]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione (38 mg, 0.04 mol, yield 98 %) was obtained in the form of a colorless oil.

[001286] Step 5: Preparation of 5-(2-(2-((1-((2R)-2-((6-chloro-4-(4-(2,2-dihydroxyacetyl)piperazin-1-yl) -8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3- yl)isoindoline-1,3-dione

[001287] To a solution of 2,2-dihydroxyacetic acid (37 mg, 0.4 mmol, 10 eq.), 5-[2-[2-[[1-[(2R)-2-[6-chloro-8 -fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]ethoxy]-2-(2,6- dioxo-3-piperidyl)isoindolin-1,3-dione (38 mg, 0.04 mmol, 1 eq., hydrochloride) and o-(7-azabenzotriazol-1-yl)-n,n,n',n '-tetramethyluronium hexafluorophosphate (61 mg, 0.16 mmol, 4 eq.) was added to diisopropylethylamine (16 mg, 0.12 mmol, 0.02 mL, 3 eq.) in N,N-dimethylformamide (2 mL), the mixture was stirred at a temperature of 25°C for 16 hours. The reaction mixture was quenched with water (30 ml) and extracted with ethyl acetate (20 ml x 2), the combined organic layers were washed with brine (30 ml), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by semipreparative reverse phase HPLC. Compound 5-[2-[2-[[1-[(2R)-2-[6-chloro-4-[4-(2,2-dihydroxyacetyl)piperazin-1-yl]-8-fluoro-7- (3-hydroxy-1-naphthyl)quinazolin-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]ethoxy]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione (7.4 mg, 0.007 mmol, 18% yield, 96% purity) was obtained as a white solid. LC/MS (ESI) m/z: 984.3 [M+l] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.08 (s, 1H), 10.01 (s, 1H), 8.13 (s, 1H), 8.00 (s, 1H), 7.79 (d, J=8.3 Hz, 1H), 7.46-7.38 (m, 2H), 7.34-7.30 (m, 1H), 7.27 (s, 1H) , 7.23-7.14 (m, 2H), 7.05 (dd, J=2.4, 5.8 Hz, 1H), 6.36 (d, J=7.3 Hz, 1H), 5.42-5.34 (m, 1H), 5.09 (dd, J=5.4, 12.8 Hz, 1H), 4.27 (s, 2H), 3.92-3.80 ( m, 5H), 3.78-3.69 (m, 4H), 3.54 (d, J=2.6 Hz, 3H), 3.48 (dd, J=5.1, 10.9 Hz , 4H), 2.93-2.81 (m, 3H), 2.63-2.51 (m, 5H), 2.23 (s, 2H), 2.08-1.99 (m, 1H ), 1.73 (s, 2H), 1.37-1.24 (m, 5H).

[001288] Example of the synthesis of (2S,4R)-N-(2-(2-((1-((R)-2-((4-(4-acryloylpiperazin-1-yl)-7-(3-hydroxynaphthalene -1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)-4-(4-methylthiazol- 5-yl)benzyl)-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide (typical compound 436)

[001289] Step 1: Preparation of tert-butyl 4-(2-(2-(((2S,4R)-4-hydroxy-1-(3-methyl-2-(3-methylisoxazol-5-yl)butanoyl) pyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)piperidine-1-carboxylate

[001290] To a solution of tert-butyl 4-[2-(p-tolylsulfonyloxy)ethoxy]piperidine-1-carboxylate (361 mg, 0.90 mmol, 1.5 eq.) and (2S,4R)-4-hydroxy -N-[[2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]methyl]-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidin-2-carboxamide (300 mg, 0.60 mmol, 1 eq.) in acetonitrile (6 ml) was added potassium carbonate (166 mg, 1.20 mmol, 2 eq.). The mixture was stirred at 80°C for 12 hours. Water (30 ml) was added. The aqueous phase was extracted with ethyl acetate (30 ml x 2). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The crude product was purified by semipreparative reverse phase HPLC. Compound tert-butyl 4-[2-[2-[[[(2S,4R)-4-hydroxy-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidin-2-carbonyl ]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]ethoxy]piperidine-1-carboxylate (320 mg, 0.44 mmol, 73% yield) was obtained as a white solid. LC/MS (ESI) m/z: 626.2 [M-100] ⁺ .

[001291] Step 2: Preparation of tert-butyl 4-(2-(2-(((2S,4R)-4-hydroxy-1-(R)-3-methyl-2-(3-methylisoxazol-5-yl )butanoyl)pyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)piperidine-1-carboxylate

[001292] Tert-butyl 4-[2-[2-[[[(2S,4R)-4-hydroxy-1-[3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2 -carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]ethoxy]piperidine-1-carboxylate (320 mg, 0.44 mmol, 1 eq.) was resolved using chiral SFC. The mixture was concentrated under reduced pressure to obtain the product. Compound tert-butyl 4-[2-[2-[[[(2S,4R)-4-hydroxy-1-[(2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine -2-carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]ethoxy]piperidine-1-carboxylate (140 mg, 0.19 mmol, 85% yield, 97% purity) was obtained as colorless oil.

[001293] Step 3: Preparation of (2S,4R)-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)-R-(4-(4- methylthiazol-5-yl)-2-(2-(piperidin-4-yloxy)ethoxy)benzyl)pyrrolidine-2-carboxamide

[001294] To a solution of tert-butyl 4-[2-[2-[[[(2S,4R)-4-hydroxy-1-[(2R)-3-methyl-2-(3-methylisoxazol-5-yl )butanoyl]pyrrolidin-2-carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]ethoxy]piperidine-1-carboxylate (140 mg, 0.19 mmol, 1 eq.) in dichloromethane ( 3 ml) hydrochloric acid/dioxane (4 M, 3 ml, 62.22 eq.) was added and the mixture was stirred at 25°C for 2 hours. The mixture was concentrated under reduced pressure to obtain a residue. Compound (2S,4R)-4-hydroxy-1-[(2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]-N-[[4-(4-methylthiazol-5-yl) )-2-[2-(4-piperidyloxy)ethoxy]phenyl]methyl]pyrrolidine-2-carboxamide (120 mg, 0.18 mmol, 94%o yield, hydrochloride) was obtained as a colorless oil. LC/MS (ESI) m/z: 626.2 [M+1] ⁺ .

[001295] Step 4: Preparation of (2S,4R)-N-(2-(2-((1-((R)-2-((4-(4-acryloylpiperazin-1-yl)-7-(3 -hydroxynaphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)-4-(4- methylthiazol-5-yl)benzyl)-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide

[001296] To a mixture of (2R)-2-[[7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)-6,8-dihydro-5H-pyrido [3,4-d]pyrimidin-2-yl]oxy]propanal (50 mg, 0.10 mmol, 1 eq.) in dichloromethane (3 ml) and methanol (1 ml) was added sodium acetate (8 mg, 0. 1 mmol, 1 eq.), stirred at 25°C for 15 minutes, then acetic acid (6 mg, 0.1 mmol, 1 eq.) and (2S,4R)-4-hydroxy-1-[(2R )-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]-N-[[4-(4-methylthiazol-5-yl)-2-[2-(4-piperidyloxy)ethoxy]phenyl ]methyl]pyrrolidine-2-carboxamide (64 mg, 0.1 mmol, 1 eq., hydrochloride) was added and stirred at 0°C for 15 minutes, then borane;2-methylpyridine (44 mg, 0.41 mmol, 4 eq.) was added at 0°C, then the reaction mixture was stirred at 25°C for 12 hours. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by semipreparative reverse phase HPLC. Compound (2S,4R)-4-hydroxy-N-[[2-[2-[[1-[(2R)-2-[[7-(3-hydroxy-1-naphthyl)-4-(4- prop-2-enoylpiperazin-1-yl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]-4-( 4-methylthiazol-5-yl)phenyl]methyl]-1-[(2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (12.4 mg, 0. 01 mmol, yield 10%, purity 95%) was obtained as a pink solid. LC/MS (ESI) m/z: 1097.5 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 9.66 (s, 1H), 8.97 (s, 1H), 8.28 (s, 1H), 8.00 (d, J=7 .7 Hz, 1H), 7.66 (d, J=8.8 Hz, 1H), 7.37 (d, J=6.6 Hz, 1H), 7.26 (s, 1H), 7. 04-6.95 (m, 2H), 6.87-6.76 (m, 3H), 6.21-6.12 (m, 2H), 5.72 (d, J=12.3 Hz, 1H), 5.20 (s, 1H), 4.35-4.04 (m, 8H), 3.85-3.63 (m, 8H), 3.51 (s, 14H), 2.90 -2.73 (m, 2H), 2.27-2.09 (m, 9H), 2.04-1.77 (m, 5H), 1.40 (s, 2H), 1.25 (d , J=6.l Hz, 3H), 0.96 (d, J=6.6 Hz, 2H), 0.79 (d, J=6.6 Hz, 2H), 0.69 (d, J =6.2 Hz, 1H), 0.57 (d, J=6.7 Hz, 1H).

[001297] Example of synthesis of (S)-N-((S)-2-((S)-2-(4-(4-(2-(2-(2-((1-((R)-2 -((4-(4-acryloylpiperazin-1-yl)-7-(3-hydroxynaphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy )propyl)piperidin-4-yl)oxy)ethoxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)-2-(methylamino) propanamide (typical compound 447)

[001298] Step 1: Preparation of 2-(2-(2-(piperidin-4-yloxy)ethoxy)ethoxy)ethan-1-ol

[001299] To a mixture of tert-butyl 4-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]piperidine-1-carboxylate (950 mg, 2.85 mmol, 1 eq.) in dichloromethane (15 ml) was added hydrogen chloride/methanol (4 M, 0.7 ml, 1 eq.). The mixture was stirred at 25°C for 1 hour. The mixture was concentrated in vacuo to give the product 2-[2-[2-(4-piperidyloxy)ethoxy]ethoxy]ethanol (760 mg, 2.82 mmol, 99% yield, hydrochloride) as a yellow oil. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 9.43-8.89 (m, 2H), 3.60-3.51 (m, 2H), 3.49-3.32 (m, 7H), 3.14 (s, 3H), 3.12-2.99 (m, 2H), 2.95-2.80 (m, 2H), 2.43-2.21 (m, 1H) , 2.00-1.82 (m, 2H), 1.74-1.54 (m, 2H).

[001300] Step 2: Preparation of benzyl 4-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)piperidine-1-carboxylate

[001301] To a mixture of 2-[2-[2-(4-piperidyloxy)ethoxy]ethoxy] ethanol (760 mg, 2.82 mmol, 1 eq., hydrochloride) in water (5 ml) and tetrahydrofuran (10 ml) sodium bicarbonate (710 mg, 8.45 mmol, 3 eq.) was added. The mixture was stirred at 25°C for 30 minutes. Benzyl chloroformate (720 mg, 4.23 mmol, 0.6 mL, 1.5 eq.) was then added to the mixture. The mixture was stirred at 25°C for 12 hours. The mixture was then diluted with water (30 ml). The mixture was then extracted with ethyl acetate (30 ml x 3). The combined organic layers were washed with water (50 ml x 2) and brine (50 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the product benzyl 4-[2-[2-(2-hydroxyethoxy)ethoxy] ethoxy]piperidine-1-carboxylate (770 mg, 2.10 mmol) as a colorless oil. LC/MS (ESI) m/z: 390.2 [M+23] ⁺ .

[001302] Step 3: Preparation of benzyl 4-(2-(2-(2-(tosyloxy)ethoxy)ethoxy)ethoxy)piperidine-1-carboxylate

[001303] To a mixture of benzyl 4-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]piperidine-1-carboxylate (770 mg, 2.10 mmol, 1 eq.) in dichloromethane (15 ml) was added p- toluenesulfonyl chloride (479 mg, 2.51 mmol, 1.2 eq.) and triethylamine (424 mg, 4.19 mmol, 0.5 ml, 2 eq.). The mixture was stirred at 25°C for 12 hours. The mixture was then concentrated in vacuo to obtain a residue. The residue was purified by chromatography on silica gel (petroleum ether: ethyl acetate in a ratio of 10:1 to 1:1). Benzyl 4-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (100 mg, 0.19 mmol, 10% yield) was obtained as a colorless oil. LC/MS (ESI) m/z: 544.3 [M+23] ⁺ .

[001304] Step 4: Preparation of benzyl 4-(2-(2-(2-((4-(2-((S)-1-((S)-2-((S)-2-((tert -butoxycarbonyl)(methyl)amino)propanamido)-2-cyclohexylacetyl)pyrrolidin-2-yl)thiazol-4-yl)naphthalene-1-yl)oxy)ethoxy)ethoxy)ethoxy)piperidine-1-carboxylate

[001305] To a mixture of benzyl 4-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (100 mg, 0.19 mmol, 1 eq.) and tert-butyl N-[(1S)-2-[[(1S)-1-cyclohexyl-2-[(2S)-2-[4-(4-hydroxy-1-naphthyl)thiazol-2-yl]pyrrolidin-1- yl]-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]-N-methyl-carbamate (119 mg, 0.19 mmol, 1 eq.) in acetonitrile (5 ml) added potassium carbonate (52 mg, 0.38 mmol, 2 eq.). The mixture was stirred at 90°C for 12 hours. The mixture was diluted with water (30 ml). The mixture was then extracted with ethyl acetate (30 ml x 3). The combined organic layers were washed with water (50 ml x 2) and brine (50 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to leave a residue. The residue was purified by preparative TLC (ethyl acetate). Benzyl compound 4-[2-[2-[2-[[4-[2-[(2S)-1-[(2S)-2-[[(2S)-2-[tert-butoxycarbonyl(methyl)amino ]propanoyl]amino]-2-cyclohexyl-acetyl]pyrrolidin-2-yl]thiazol-4-yl]-1-naphthyl]oxy]ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (120 mg) was obtained as green oils LC/MS (ESI) m/z: 970.6 [M+1] ⁺ .

[001306] Step 5: Preparation of tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-oxo-2-((S)-2-(4-(4-(2-( 2-(2-(piperidin-4-yloxy)ethoxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)ethyl)amino)-1-oxopropan-2-yl) methyl)carbamate

[001307] To a mixture of benzyl 4-[2-[2-[2-[[4-[2-[(2S)-1-[(2S)-2-[[(2S)-2-[tert-butoxycarbonyl (methyl)amino]propanoyl]amino]-2-cyclohexyl-acetyl]pyrrolidin-2-yl]thiazol-4-yl]-1-naphthyl]oxy]ethoxy]ethoxy]ethoxy]piperidine-1-carboxylate (120 mg, 0.12 mmol, 1 eq.) to trifluoroethanol (10 ml) was added a palladium-activated carbon catalyst (30 mg, 10% purity) under a nitrogen atmosphere. The mixture was degassed and charged with hydrogen 3 times. It was then stirred at 25°C for 2 hours. The mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC. Tert-butyl compound N-[(1S)-2-[[(1S)-1-cyclohexyl-2-oxo-2-[(2S)-2-[4-[4-[2-[2-[2 -(4-piperidyloxy)ethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]ethyl]amino]-1-methyl-2-oxo-ethyl]-N-methyl-carbamate (35 mg, 0.04 mmol, 32% yield, formate) was obtained as a white solid. LC/MS (ESI) m/z: 836.6 [M+1] ⁺ .

[001308] Step 6: Preparation of tert-butyl ((S)-1-(((S)-2-((S)-2-(4-(4-(2-(2-(2-((1 -((R)-2-((4-(4-acryloylpiperazin-1-yl)-7-(3-hydroxynaphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d] pyrimidin-2-yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl )amino)-1-oxopropan-2-yl)(methyl)carbamate

[001309] To a mixture of tert-butyl N-[(1S)-2-[[(1S)-1-cyclohexyl-2-oxo-2-[(2S)-2-[4-[4-[2-[ 2-[2-(4-piperidyloxy)ethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]ethyl]amino]-1-methyl-2-oxo-ethyl]-N -methyl-carbamate (45 mg, 0.05 mmol, 1 eq., formate) in methanol (2 ml) and dichloromethane (2 ml) was added sodium acetate (12 mg, 0.15 mmol, 3 eq.). The borane complex was then added to the mixture with stirring; 2-methylpyridine (27 mg, 0.25 mmol, 5 eq.). The mixture was then stirred at 25°C for 20 minutes. Then acetic acid (6 mg, 0.10 mmol, 2 eq.) and (2R)-2-[[7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazine-1- yl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxy]propanal (24 mg, 0.05 mmol, 1 eq.) was added at 0°C. The reaction mixture was then stirred at 25°C for 12 hours. The mixture was then concentrated in vacuo to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol 9:1). Tert-butyl N-[(1S)-2-[[(1S)-1-cyclohexyl-2-[(2S)-2-[4-[4-[2-[2-[2-[[1- [(2R)-2-[[7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)-6,8-dihydro-5H-pyrido[3,4 -d]pyrimidin-2-yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-2-oxo-ethyl] amino]-1-methyl-2-oxo-ethyl]-N-methyl-carbamate (26 mg, 0.02 mmol, 39% yield) was obtained as a pink oil. LC/MS (ESI) m/z: 1307.5 [M+1] ⁺ .

[001310] Step 7: Preparation of (S)-N-((S)-2-((S)-2-(4-(4-(2-(2-(2-((1-((R) -2-((4-(4-acryloylpiperazin-1-yl)-7-(3-hydroxynaphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl )oxy)propyl)piperidin-4-yl)oxy)ethoxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)-2-( methylamino)propanamide

[001311] To a mixture of tert-butyl N-[(1S)-2-[[(1S)-1-cyclohexyl-2-[(2S)-2-[4-[4-[2-[2-[2 -[[1-[(2R)-2-[[7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)-6,8-dihydro-5H- pyrido[3,4-d]pyrimidin-2-yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-2 -oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]-N-methyl-carbamate (26 mg, 0.02 mmol, 1 eq.) in dichloromethane (2.5 ml) was added trifluoroacetic acid (3 .38 mmol, 0.25 ml). The mixture was stirred at 25°C for 30 minutes. Then the mixture was adjusted to pH=7-8 with a solution of sodium bicarbonate, then the mixture was filtered. The filtrate was purified by preparative HPLC. Compound (2S)-N-[(1S)-1-cyclohexyl-2-[(2S)-2-[4-[4-[2-[2-[2-[[1-[(2R)-2 -[[7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2 -yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-2-oxo-ethyl]-2-(methylamino) propanamide (12.4 mg, 0.01 mmol, 48% yield, 95% purity, formate) was obtained as a pink solid. LC/MS (ESI) m/z: 604.4 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.29-8.20 (m, 4H), 8.07-7.96 (m, 2H), 7.71-7.62 (m, 2H), 7.59-7.57 (d, J=8.0 Hz, 1H), 7.56-7.49 (m, 2H), 7.42-7.35 (m, 1H), 7 ,30-7.20 (m, 1H), 7.03-7.01 (d, J=8.0 Hz, 1H), 6.88-6.74 (m, 3H), 6.19-6 .10 (m, 1H), 5.75-5.68 (m, 1H), 5.48-5.39 (m, 1H), 5.23-5.11 (m, 1H), 4.55 -4.47 (m, 1H), 4.34-4.27 (m, 2H), 4.05 (s, 2H), 3.93-3.88 (m, 2H), 3.82-3 .77 (m, 2H), 3.73-3.62 (m, 8H), 3.58-3.55 (m, 3H), 3.26-3.17 (m, 6H), 3.14 -3.05 (m, 2H), 2.92-2.83 (m, 2H), 2.79-2.68 (m, 2H), 2.31-2.24 (m, 2H), 2 .23-2.17 (m, 4H), 2.14-1.99 (m, 5H), 1.88-1.47 (m, 10H), 1.39-1.28 (m, 2H) , 1.23-1.21 (d, J=8.0 Hz, 3H), 1.16-0.94 (m, 9H).

[001312] Example of synthesis of (S)-N-((S)-2-((S)-2-(4-(4-(2-(2-((1-((R)-2-(( 4-(4-acryloylpiperazin-1-yl)-7-(3-hydroxynaphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)propyl) piperidin-4-yl)oxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)-2-(methylamino)propanamide (typical compound 446 )

[001313] Step 1: Preparation of 2-(2-(piperidin-4-yloxy)ethoxy)ethan-1-ol

[001314] To a solution of tert-butyl 4-[2-(2-hydroxyethoxy)ethoxy]sheridine-1-carboxylate (1 g, 3.46 mmol, 1 eq.) in hydrochloride/methanol (4 M, 3 mL, 3 .47 eq.), the mixture was stirred at 25°C for 1 hour. The mixture was filtered and concentrated under reduced pressure to give the product 2-[2-(4-piperidyloxy)ethoxy] ethanol (600 mg, hydrochloride) as a white solid.

[001315] Step 2: Preparation of benzyl 4-(2-(2-hydroxyethoxy)ethoxy)piperidine-1-carboxylate

[001316] To a solution of 2-[2-(4-piperidyloxy)ethoxy]ethanol (600 mg, 3.17 mmol, 1 eq.) in dichloromethane (6 ml) was added benzyl carbonochloridate (568 mg, 3.33 mmol, 0 .5 ml, 1.05 eq.) and triethylamine (481 mg, 4.76 mmol, 0.7 ml, 1.5 eq.), the mixture was stirred at 25°C for 2 hours. The reaction mixture was extracted with dichloromethane (20 ml x 2) and hydrochloric acid solution (1 M) (30 ml). The combined organic layers were washed with brine (50 ml), dried over sodium sulfate, concentrated under reduced pressure to give the product benzyl 4-[2-(2-hydroxyethoxy)ethoxy] piperidine-1-carboxylate (625 mg) as a colorless oil. LC/MS (ESI) m/z: 324.2 [M+1] ⁺ .

[001317] Step 3: Preparation of benzyl 4-(2-(2-(tosyloxy)ethoxy)ethoxy)piperidine-1-carboxylate

[001318] To a solution of benzyl 4-[2-(2-hydroxyethoxy)ethoxy]piperidine-1-carboxylate (625 mg, 1.93 mmol, 1 eq.) in dichloromethane (10 ml) was added triethylamine (587 mg, 5. 80 mmol, 0.8 ml, 3 eq.), then p-toluenesulfonyl chloride (553 mg, 2.90 mmol, 1.5 eq.) was added to the mixture, the mixture was stirred at 25°C for 16 hours. The reaction mixture was quenched with water (30 ml), and extracted with dichloromethane (50 ml x 2), the combined organic layers were washed with brine (30 ml), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (petroleum ether:ethyl acetate 30:1 to 1:1). The compound benzyl 4-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]piperidine-1carboxylate (650 mg, 1.36 mmol, 70% yield) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CD ₃ OD) δ 7.82-7.76 (m, 2H), 7.42 (d, J=8.0 Hz, 2H), 7.38-7.28 ( m, 5H), 5.11 (s, 2H), 4.17-4.13 (m, 2H), 3.79-3.69 (m, 2H), 3.68-3.61 (m, 2H), 3.58-3.49 (m, 5H), 3.22 (s, 2H), 2.47-2.40 (m, 3H), 1.87-1.77 (m, 2H) , 1.47 (tdd, J=4.4, 8.4, 12.8 Hz, 2H).

[001319] Step 4: Preparation of benzyl 4-(2-(2-((4-(2-((S)-1-((S)-2-((S)-2-((tert-butoxycarbonyl) (methyl)amino)propanamido)-2-cyclohexylacetyl)pyrrolidin-2-yl)thiazol-4-yl)naphthalene-1-yl)oxy)ethoxy)ethoxy)piperidine-1-carboxylate

[001320] To a solution of benzyl 4-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]piperidine-1-carboxylate (231 mg, 0.48 mmol, 1 eq.), tert-butyl N-[(1S )-2-[[(1S)-1-cyclohexyl-2-[(2S)-2-[4-(4-hydroxy-1-naphthyl)thiazol-2-yl]pyrrolidin-1-yl]-2- oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]-N-methyl-carbamate (300 mg, 0.48 mmol, 1 eq.) in acetonitrile (5 ml) was added potassium carbonate (167 mg, 1 .21 mmol, 2.5 eq.), the mixture was stirred at 80°C for 6 hours. The mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol 10:1). Benzyl compound 4-[2-[2-[[4-[2-[(2S)-1-[(2S)-2-[[(2S)-2-[tert-butoxycarbonyl(methyl)amino]propanoyl] amino]-2-cyclohexyl-acetyl]pyrrolidin-2-yl]thiazol-4-yl]-1-naphthyl]oxy]ethoxy]ethoxy]piperidine-1-carboxylate (400 mg, 0.36 mmol, 74% yield, purity 83%) was obtained as a yellow oil. LC/MS (ESI) m/z: 926.5 [M+1] ⁺ .

[001321] Step 5: Preparation of tert-butyl ((S)-1-(((S)-1-cyclohexyl-2-oxo-2-((S)-2-(4-(4-(2-( 2-(piperidin-4-yloxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)ethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate

[001322] To a solution of benzyl 4-[2-[2-[[4-[2-[(2S)-1-[(2S)-2-[[(2S)-2-[tert-butoxycarbonyl(methyl) amino]propanoyl]amino]-2-cyclohexyl-acetyl]pyrrolidin-2-yl]thiazol-4-yl]-1-naphthyl]oxy]ethoxy]ethoxy]piperidine-1-carboxylate (350 mg, 0.38 mmol, 1 eq.) in trifluoroethanol (20 ml), a palladium-on-activated carbon catalyst (100 mg, 0.38 mmol, 10% purity, 1 eq.) was added under a nitrogen atmosphere. The suspension was degassed in vacuum and purged with hydrogen several times. The mixture was stirred under a hydrogen atmosphere [0.1 MPa (15 psi)] at 25°C for 2 hours. The mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by semipreparative reverse phase HPLC. Tert-butyl compound N-[(1S)-2-[[(1S)-1-cyclohexyl-2-oxo-2-[(2S)-2-[4-[4-[2-[2-(4 -piperidyloxy)ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]ethyl]amino]-1-methyl-2-oxo-ethyl]-M-methyl-carbamate (111 mg, 0 .14 mmol, yield 37%) was obtained as a colorless oil.

[001323] Step 6: Preparation of tert-butyl ((S)-1-(((S)-2-((S)-2-(4-(4-(2-(2-((1-(( R)-2-((4-(4-acryloylpiperazin-1-yl)-7-(3-hydroxynaphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2 -yl)oxy)propyl)piperidin-4-yl)oxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)amino)-1 -oxopropan-2-yl)(methyl)carbamate

[001324] To a solution of tert-butyl N-[(1S)-2-[[(1S)-1-cyclohexyl-2-oxo-2-[(2S)-2-[4-[4-[2-[ 2-(4-piperidyloxy)ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]ethyl]amino]-1-methyl-2-oxo-ethyl]-N-methyl-carbamate ( 81 mg, 0.1 mmol, 1 eq.) in methanol (3 ml) was added sodium acetate (17 mg, 0.21 mmol, 2 eq.), then borane;2-methylpyridine (44 mg, 0.41 mmol, 4 eq.) in dichloromethane (1 ml) was added at 0°C for 0.5 hour. (2R)-2-[[7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)-6,8-dihydro-5H-pyrido[3,4- d]pyrimidin-2-yl]oxy]propanal (50 mg, 0.1 mmol, 1 eq.) and acetic acid (0.62 mg, 0.01 mmol, 0.1 eq.) were added to the mixture and stirred at temperature 25°C for 11.5 hours. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol 10:1). Tert-butyl compound N-[(1S)-2-[[(1S)-1-cyclohexyl-2-[(2S)-2-[4-[4-[2-[2-[[1-[( 2R)-2-[[7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)-6,8-dihydro-5H-pyrido[3,4-d ]pyrimidin-2-yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-2-oxo-ethyl]amino]-1 -Methyl-2-oxo-ethyl]-N-methyl-carbamate (60 mg, 0.05 mmol, 46% yield) was obtained as a colorless oil.

[001325] Step 7: Preparation of (S)-N-((S)-2-((S)-2-(4-(4-(2-(2-((1-((R)-2- ((4-(4-acryloylpiperazin-1-yl)-7-(3-hydroxynaphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy) propyl)piperidin-4-yl)oxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-1-cyclohexyl-2-oxoethyl)-2-(methylamino)propanamide

[001326] To a solution of tert-butyl N-[(1S)-2-[[(1S)-1-cyclohexyl-2-[(2S)-2-[4-[4-[2-[2-[[ 1-[(2R)-2-[[7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)-6,8-dihydro-5H-pyrido[3 ,4-d]pyrimidin-2-yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-2-oxo-ethyl] amino]-1-methyl-2-oxo-ethyl]-N-methyl-carbamate (60 mg, 0.05 mol, 1 eq.) in dichloromethane (2 ml) was added trifluoroacetic acid (308 mg, 2.70 mmol, 0.2 ml, 56.89 eq.), the mixture was stirred at 25°C for 10 minutes. The reaction was poured into an aqueous sodium bicarbonate solution and the pH was adjusted to approximately 7~8. The reaction mixture was quenched with water (30 ml) and extracted with ethyl acetate (20 ml x 2), the combined organic layers were washed with brine (30 ml), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by semipreparative reverse phase HPLC. Compound (2S)-N-[(1S)-1-cyclohexyl-2-[(2S)-2-[4-[4-[2-[2-[[1-[(2R)-2-[[ 7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl] oxy]propyl]-4-piperidyl]oxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-2-oxo-ethyl]-2-(methylamino)propanamide (6.6 mg, 0.01 mmol, yield 11%, purity 95%) was obtained as a pink solid. LC/MS (ESI) m/z: 1185.6 [M+23] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.33 (s, 1H), 8.28-8.21 (m, 2H), 8.04-7.90 (m, 2H), 7 .71-7.63 (m, 2H), 7.60-7.48 (m, 3H), 7.38 (t, J=7.4 Hz, 1H), 7.26 (t, J=7 ,2 Hz, 1H), 7.02 (d, J=8.1 Hz, 1H), 6.86-6.75 (m, 3H), 6.14 (dd, J=2.4, 16, 7 Hz, 1H), 5.71 (d, J=12.7 Hz, 1H), 5.42 (d, J=5.6 Hz, 1H), 5.24-5.14 (m, 1H) , 4.57-4.48 (m, 1H), 4.31 (s, 2H), 4.05 (s, 2H), 3.91 (s, 2H), 3.80 (t, J=7 ,1 Hz, 2H), 3.71-3.63 (m, 6H), 3.56 (d, J=5.4 Hz, 3H), 2.99 (q, J=7.1 Hz, 3H ), 2.88 (s, 2H), 2.75-2.65 (m, 3H), 2.35-2.14 (m, 10H), 2.12-2.02 (m, 4H), 1.81-1.53 (m, 10H), 1.36 (s, 2H), 1.23 (d, J=6.1 Hz, 3H), 1.13-1.02 (m, 7H) .

[001327]

[001328] Example of synthesis of (2S,4R)-1-((S)-2-(2-(2-(2-(((3R,5S)-5-(((4-((S)-4 -acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl )-1-methylpyrrolidin-3-yl)oxy)ethoxy)ethoxy)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl) )phenyl)ethyl)pyrrolidine-2-carboxamide (typical compound 500)

[001329] Step 1: Preparation of tert-butyl (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-ethoxy-2-oxoethoxy) ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin- 1-carboxylate

[001330] To a mixture of tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-(2-hydroxyethoxy)ethoxy]-1-methyl-pyrrolidine- 2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (300 mg, 0.43 mmol , 1 eq.) in dichloromethane (10 ml) rhodium (ii) acetate dimer (9 mg, 0.042 mmol, 0.1 eq.) was added, then ethyl 2-diazoacetate (146 mg, 1.28 mmol, 3 eq.) was added at 0°C, then the reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was concentrated in vacuo. The residue was purified by preparative TLC (10:1 dichloromethane:methanol) to give tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2 -(2-ethoxy-2-oxo-ethoxy)ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3, 4-d]pyrimidin-4-yl]piperazin-1-carboxylate (111 mg, 0.14 mmol, 33% yield) as a yellow oil. LC/MS (ESI) m/z: 788.3 [M+1] ⁺ .

[001331] Step 2: Preparation of 2-(2-(2-(((3R,5S)-5-(((4-((S)-4-(tert-butoxycarbonyl)-3-(cyanomethyl)piperazine- 1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)-1-methylpyrrolidin-3-yl) hydroxy)ethoxy)ethoxy)acetic acid

[001332] To a mixture of tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-(2-ethoxy-2-oxo-ethoxy) ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin- 1-carboxylate (110 mg, 0.14 mmol, 1 eq.) in water (1 ml), and tetrahydrofuran (1 ml), and methanol (1 ml) was added lithium hydroxide monohydrate (126 mg, 3 mmol, 21.49 eq.), then the reaction mixture was stirred at a temperature of 25°C for 2 hours. Tetrahydrofuran (5 ml) and water (5 ml) were added, then the reaction mixture was adjusted to pH 2-3, the aqueous phase was extracted with dichloromethane and methanol (10:1) (20 ml x 4), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to obtain the compound 2-[2-[2-[(3R,5S)-5-[[4-[(3S)-4-tert-butoxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-7- (1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl]oxyethoxy]ethoxy]acetic acid (131 mg ) as a yellow oil.

[001333] Step 3: Preparation of tert-butyl (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-(((S)- 1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3, 3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7 ,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazine-1-carboxylate

[001334] To a mixture of (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (134 mg, 0.28 mmol, 2 eq., hydrochloride) and 2-[2-[2-[(3R,5S)-5-[[ 4-[(3S)-4-tert-butoxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine -2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl]oxyethoxy]ethoxy]acetic acid (106 mg, 0.14 mmol, 1 eq.) in N-methyl-2-pyrrolidone (4 ml) was added 1-Hydroxybenzotriazole (28 mg, 0.21 mmol, 1.5 eq.), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (40 mg, 0.21 mmol, 1.5 eq.), N,N- diisopropylethylamine (108 mg, 0.84 mmol, 0.1 ml, 6 eq.), then the reaction mixture was stirred at 20°C for 12 hours. The residue was poured into ice water (20 ml). The aqueous phase was extracted with ethyl acetate (20 ml * 3). The combined organic phase was washed with brine (20 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative TLC (10:1 dichloromethane:methanol) to give tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2 -[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl ]pyrrolidin-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl) -6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (90 mg) as a yellow oil. LC/MS (ESI) m/z: 594.0 [M/2+1] ⁺ .

[001335] Step 4: Preparation of (2S,4R)-1-((S)-2-(2-(2-(2-(((3R,5S)-5-(((4-((S) -3-(cyanomethyl)piperazin-1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)- 1-methylpyrrolidin-3-yl)oxy)ethoxy)ethoxy)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl )ethyl)pyrrolidine-2-carboxamide

[001336] To a solution of tert-butyl (2S)-2-(nianomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-[[(1S)-1- [(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2- dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3 ,4-d]pyrimidin-4-yl]piperazin-1-carboxylate (35 mg, 0.030 mmol, 1 eq.) in dichloromethane (3 ml) was added trifluoroacetic acid (462 mg, 4.05 mmol, 0.3 ml, 137.35 eq.). The mixture was stirred at 15°C for 20 minutes. The reaction mixture was concentrated in vacuo to give the compound (2S,4R)-1-[(2S)-2-[[2-[2-[2-[(3R,5S)-5-[[4-[(3S) -3-(cyanomethyl)piperazin-1-yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl- pyrrolidin-3-yl]oxyethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl ]ethyl]pyrrolidine-2-carboxamide (35 mg, 0.030 mmol, 99% yield, trifluoroacetate) as a yellow oil. LC/MS (ESI) m/z: 1086.7 [M+1] ⁺ .

[001337] Step 5: Preparation of (2S,4R)-1-((S)-2-(2-(2-(2-(((3R,5S)-5-(((4-((S) -4-acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy )methyl)-1-methylpyrrolidin-3-yl)oxy)ethoxy)ethoxy)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5 -yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001338] To the mixture (2S,4R)-1-[(2S)-2-[[2-[2-[2-[(3R,5S)-5-[[4-[(3S)-3- (cyanomethyl)piperazin-1-yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidine-3 -yl]oxyethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] pyrrolidine-2-carboxamide (30 mg, 0.025 mmol, 1 eq., trifluoroacetate) in dichloromethane (5 ml) was added with 2,6-lutidine (27 mg, 0.25 mmol, 10 eq.), then prop-2-enoyl chloride (2 mg, 0.022 mmol, 0.9 eq.) in dichloromethane (1.8 ml) was added at minus 65°C, then the reaction mixture was stirred at minus 65°C for 10 minutes. Water (10 ml) was added and stirred for 0.5 minutes. The aqueous phase was extracted with dichloromethane (20 ml x 2) and concentrated in vacuo. The residue was purified by semipreparative reverse phase HPLC, then the collected fraction was concentrated to remove most of the acetonitrile. The solution was lyophilized to give the compound (2S,4R)-1-[(2S)-2-[[2-[2-[2-[(3R,5S)-5-[[4-[(3S)-3- (cyanomethyl)-4-prop-2-enoyl-piperazin-1-yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl ]-1-methyl-pyrrolidin-3-yl]oxyethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole -5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (9.1 mg, 0.007 mmol, 30% yield, 97% purity, formate) as a white solid. LC/MS (ESI) m/z: 570.8 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.98 (s, 1H), 8.44 (br d, J=7.6 Hz, 1H), 8.30 (s, 1H), 8 .22-8.14 (m, 1H), 7.97-7.88 (m, 1H), 7.64 (d, J=8.2 Hz, 1H), 7.59-7.49 (m , 2H), 7.48-7.39 (m, 3H), 7.38-7.33 (m, 2H), 7.22 (d, J=7.2 Hz, 1H), 6.88 ( br s, 1H), 6.20 (br d, J=17.4 Hz, 1H), 5.78 (br d, J=11.6 Hz, 1H), 5.06-4.63 (m, 2H), 4.55 (br d, J=9.5 Hz, 1H), 4.45 (br t, J=8.1 Hz, 1H), 4.34-4.22 (m, 2H), 4.17-4.09 (m, 3H), 4.08-3.98 (m, 3H), 3.98-3.91 (m, 2H), 3.61 (brd, J=3.1 Hz, 2H), 3.58 (br d, J=3.9 Hz, 4H), 3.56-3.53 (m, 1H), 3.54 (br s, 2H), 3.53-3 .48 (m, 4H), 3.29 (br dd, J=6.2, 9.5 Hz, 2H), 3.24-3.17 (m, 2H), 3.07-2.90 ( m, 4H), 2.78-2.63 (m, 2H), 2.45 (s, 3H), 2.34 (s, 3H), 2.19 (br dd, J=6.0, 9 ,4 Hz, 1H), 2.10-2.00 (m, 1H), 1.92-1.72 (m, 3H), 1.37 (br d, J=7.0 Hz, 3H), 0.94 (s, 9H).

[001339] Example of the synthesis of 2-((2S)-1-acryloyl-4-(2-(((2S,4R)-4-(2-(2-(2-((2-(2,6-dioxopiperidine -3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6, 7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile (representative compound 504)

[001340] Step 1: Preparation of tert-butyl (2S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-((2-(2 ,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)- 5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazine-1-carboxylate

[001341] To a mixture of tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-1-methyl-4-[2-[2-[2-(p-tolylsulfonyloxy )ethoxy]ethoxy]ethoxy]pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-1- carboxylate (100 mg, 0.11 mmol, 1 eq.) and 2-(2,6-dioxo-3-piperidyl)-5-hydroxy-isoindoline-1,3-dione (152 mg, 0.56 mmol, 5 eq.) in N,N-dimethylformamide (4 ml) was added potassium carbonate (46 mg, 0.33 mmol, 3 eq.), then the reaction mixture was stirred at 50°C for 12 hours. The residue was poured into 0.1 M hydrochloric acid (30 ml). The aqueous phase was extracted with ethyl acetate (50 ml x 4), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative TLC (petroleum ether: ethyl acetate in the ratio 0:1). The compound tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-(2,6-dioxo-3- piperidyl)-1,3-dioxo-isoindolin-5-yl]oxyethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H -pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (70 mg, 0.07 mmol, 63% yield) was obtained as a yellow oil. LC/MS (EST) m/z: 1002.6 [M+1] ⁺ .

[001342] Step 2: Preparation of 2-((2S)-4-(2-(((2S,4R)-4-(2-(2-(2-((2-(2,6-dioxopiperidine-3 -yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7, 8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[001343] To a mixture of tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-(2,6- dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]oxyethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8 trifluoroacetic acid (1.54 g, 13.51 mmol, 1 ml, 386.70 eq.), then the reaction mixture was stirred at 20°C for 1 hour. The reaction mixture was concentrated in vacuo. Compound 2-[(2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-(2,6-dioxo-3-piperidyl)-1,3 -dioxo-isoindolin-5-yl]oxyethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4 -d]pyrimidin-4-yl]piperazin-2-yl]acetonitrile (35 mg, 0.034 mmol, 99% yield, trifluoroacetate) was obtained as a yellow oil.

[001344] Step 3: Preparation of 2-((2S)-1-acryloyl-4-(2-(((2S,4R)-4-(2-(2-(2-((2-(2,6 -dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5, 6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[001345] To a mixture of 2-[(2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-(2,6-dioxo-3-piperidyl) -1,3-dioxo-isoindolin-5-yl]oxyethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido [3,4-d]pyrimidin-4-yl]piperazin-2-yl]acetonitrile (35 mg, 0.034 mmol, 1 eq., trifluoroacetate) in dichloromethane (4 ml) was added 2,6-lutidine (37 mg, 0 .34 mmol, 10 eq.), then prop-2-enoyl chloride (3 mg, 0.03 mmol, 0.85 eq.) in dichloromethane (2 ml) was added at minus 65°C for 10 minutes. Water (10 ml) was added and stirred for 0.5 minutes. The aqueous phase was extracted with dichloromethane (20 ml x 2), then concentrated in vacuo. The residue was purified by semipreparative reverse phase HPLC. Compound 2-[(2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-(2,6-dioxo-3-piperidyl)-1,3 -dioxo-isoindolin-5-yl]oxyethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4 -d]pyrimidin-4-yl]-1-prop-2-enoyl-piperazin-2-yl]acetonitrile (6.2 mg, 0.006 mmol, 18% yield, 95% purity) was obtained as an off-white solid colors. LC/MS (ESI) m/z: 956.5 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.10 (br s, 1H), 8.37-8.06 (m, 2H), 7.96-7.77 (m, 2H), 7.63 (br s, 1H), 7.58-7.42 (m, 4H), 7.36 (br s, 1H), 7.22 (br s, 1H), 6.86 (br s, 1H), 6.19 (br d, J=17.0 Hz, 1H), 5.78 (br d, J=8.8 Hz, 1H), 5.11 (br d, J=7.9 Hz , 1H), 5.03-4.71 (m, 1H), 4.41 (br s, 1H), 4.34-4.23 (m, 3H), 4.13 (br s, 4H), 4.02 (br d, J=12.3 Hz, 4H), 3.78 (br s, 4H), 3.59 (br s, 2H), 3.54 (br s, 2H), 3.49 (br s, 4H), 3.30-3.16 (m, 1H), 3.06-2.86 (m, 6H), 2.67 (br s, 1H), 2.32 (br s, 3H), 2.16 (br s, 1H), 2.04 (br s, 1H), 1.85 (br s, 3H).

[001346] Example of the synthesis of (2S,4R)-N-(2-(2-(2-(((3R,5S)-5-(((4-((S)-4-acryloyl-3-(cyanomethyl )piperazin-1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)-1-methylpyrrolidin-3 -yl)oxy)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl) butanoyl)pyrrolidine-2-carboxamide (representative compound 506)

[001347] Step 1: Preparation of (2S,4R)-N-(2-(2-(2-(((3R,5S)-5-(((4-((S)-3-(cyanomethyl)piperazine -1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)-1-methylpyrrolidin-3-yl )oxy)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl) pyrrolidine-2-carboxamide

[001348] To a solution of tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-[[[(2S,4R) -4-hydroxy-1-[(2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidin-2-carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl) )phenoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl ]piperazine-1-carboxylate (80 mg, 0.067 mmol, 1 eq.) in dichloromethane (8 ml) was added trifluoroacetic acid (3.08 g, 27.0 mmol, 2.0 ml, 400 eq.). The mixture was stirred at 20°C for 2 hours. The reaction mixture was evaporated in vacuo to give the product, (2S,4R)-N-[[2-[2-[2-[(3R,5S)-5-[[4-[(3S)-3-(cyanomethyl) piperazin-1-yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl] oxyethoxy]ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]-4-hydroxy-1-[(2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine -2-carboxamide (73 mg) as a yellow oil.

[001349] Step 2: Preparation of (2S,4R)-N-(2-(2-(2-(((3R,5S)-5-(((4-((S)-4-acryloyl-3- (cyanomethyl)piperazin-1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)-1-methylpyrrolidine -3-yl)oxy)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5- yl)butanoyl)pyrrolidine-2-carboxamide

[001350] To a solution of (2S,4R)-N-[[2-[2-[2-[(3R,5S)-5-[[4-[(3S)-3-(cyanomethyl)piperazine-1- yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl]oxyethoxy]ethoxy] -4-(4-methylthiazol-5-yl)phenyl]methyl]-4-hydroxy-1-[(2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (73 mg, 0.067 mmol, 1 eq.) and 2,6-lutidine (230 mg, 2.15 mmol, 32 eq.) in a mixed solvent of dichloromethane (8 ml) and N,N-dimethylformamide (1 ml) were added prop-2-enoyl chloride (6.1 mg, 0.067 mmol, 1 eq.) in dichloromethane (0.2 ml) under nitrogen. The mixture was stirred at minus 65°C for 10 minutes. The reaction mixture was quenched with water (10 ml) before heating to 25°C, then extracted with dichloromethane (20 ml x 3). The combined organic layers were combined and evaporated in vacuo to leave a residue. The residue was purified by preparative HPLC. Product (2S,4R)-N-[[2-[2-[2-[(3R,5S)-5-[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoyl -piperazin-1-yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl ]oxyethoxy]ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]-4-hydroxy-1-[(2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl] Pyrrolidine-2-carboxamide (44.1 mg, 0.034 mmol, 50% yield, 96% purity, trifluoroacetate) was obtained as a light yellow solid. LC/MS (ESI) m/z: 1136.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.98 (s, 1H), 8.33-8.30 (m, 1H), 8.19-8.17 (m, 1H), 7 .95-7.92 (m, 1H), 7.65 (d, J=8.0 Hz, 1H), 7.55-7.53 (m, 2H), 7.47 (t, J=7 .6 Hz, 1H), 7.32-7.21 (m, 2H), 7.08-6.97 (m, 2H), 6.87 (s, 1H), 6.22-6.18 ( m, 2H), 5.81-5.78 (m, 1H), 5.00-4.76 (m, 1H), 4.62-4.59 (m, 1H), 4.48-4, 41 (m, 3H), 4.33-4.08 (m, 12H), 3.98-3.88 (m, 1H), 3.85 (d, J=8.4 Hz, 1H), 3 .83-3.71 (m, 3H), 3.65-3.59 (m, 6H), 3.55-3.44 (m, 1H), 3.41-3.33 (m, 1H) , 3.31-3.14 (m, 4H), 3.08-3.29 (m, 7H), 2.46-2.44 (m, 3H), 2.37-2.23 (m, 2H), 2.21-2.13 (m, 3H), 2.08-1.86 (m, 2H), 1.00-0.55 (m, 6H).

[001351] Example of the synthesis of (2S,4R)-N-(2-(2-(2-(((3R,5S)-5-(((4-((S)-4-acryloyl-3-(cyanomethyl )piperazin-1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)-1-methylpyrrolidin-3 -yl)oxy)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((S)-3-methyl-2-(3-methylisoxazol-5-yl) butanoyl)pyrrolidine-2-carboxamide (representative compound 507)

[001352] Step 1: Preparation of (2S,4R)-N-(2-(2-(2-(((3R,5S)-5-(((4-((S)-3-(cyanomethyl)piperazine -1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)-1-methylpyrrolidin-3-yl )oxy)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl) pyrrolidine-2-carboxamide

[001353] To a solution of tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-[[[(2S,4R) -4-hydroxy-1-[(2S)-3-methyl-2-(3-methylisoxanol-5-yl)butanoyl]pyrrolidin-2-carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl )phenoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl ]piperazine-1-carboxylate (60 mg, 0.051 mmol, 1 eq.) in dichloromethane (6 ml) was added trifluoroacetic acid (2.31 g, 20.26 mmol, 1.5 ml, 400 eq.). The mixture was stirred at 20°C for 2 hours. The mixture was evaporated in vacuo to give the product (2S,4R)-N-[[2-[2-[2-[(3R,5S)-5-[[4-[(3S)-3-(cyanomethyl)piperazine- 1-yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl]oxyethoxy] ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]-4-hydroxy-1-[(2S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidin-2 -carboxamide (50 mg) as a light yellow oil.

[001354] Step 2: Preparation of (2S,4R)-N-(2-(2-(2-(((3R,5S)-5-(((4-((S)-4-acryloyl-3- (cyanomethyl)piperazin-1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)-1-methylpyrrolidine -3-yl)oxy)ethoxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxy-1-((S)-3-methyl-2-(3-methylisoxazol-5- yl)butanoyl)pyrrolidine-2-carboxamide

[001355] To a solution of (2S,4R)-N-[[2-[2-[2-[(3R,5S)-5-[[4-[(3S)-3-(cyanomethyl)piperazine-1- yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl]oxyethoxy]ethoxy] -4-(4-methylthiazol-5-yl)phenyl]methyl]-4-hydroxy-1-[(2S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (50 mg, 0.046 mmol, 1 eq.) and 2,6-lutidine (184 mg, 1.72 mmol, 0.2 mL, 37 eq.) in a mixed solvent of dichloromethane (4 mL) and N,N-dimethylformamide (0.5 ml) prop-2-enoyl chloride (3.7 mg, 0.042 mmol, 0.9 eq.) in dichloromethane (0.2 ml) under nitrogen was added. The mixture was stirred at minus 65°C for 10 minutes. The mixture was quenched with water (10 ml) and extracted with dichloromethane (20 ml x 3). The organic layers were combined and evaporated in vacuo to leave a residue (50 mg). The residue was purified by preparative HPLC. Product (2S,4R)-N-[[2-[2-[2-[(3R,5S)-5-[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoyl -piperazin-1-yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl ]oxyethoxy]ethoxy]-4-(4-methylthiazol-5-yl)phenyl]methyl]-4-hydroxy-1-[(2S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl] Pyrrolidine-2-carboxamide (28.5 mg, 0.024 mmol, 52% yield, 96% purity), trifluoroacetate) was obtained as an off-white solid. LC/MS (ESI) m/z: 1136.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.99 (s, 1H), 8.65-8.37 (m, 1H), 8.20-8.17 (m, 1H), 7 .95-7.93 (m, 1H), 7.67 (d, J=8.0 Hz, 1H), 7.56-7.54 (m, 2H), 7.47 (t, J=7 .6 Hz, 1H), 7.36 (d, J=7.6 Hz, 1H), 7.24-7.22 (m, 1H), 7.06-7.00 (m, 2H), 6 .88 (s, 1H), 6.23-6.18 (m, 2H), 5.81-5.78 (m, 1H), 4.99-4.78 (m, 1H), 4.63 -4.59 (m, 1H), 4.48-4.17 (m, 12H), 4.10-4.07 (m, 3H), 3.96-3.92 (m, 1H), 3 .82-3.73 (m, 5H), 3.67-3.55 (m, 5H), 3.46-3.37 (m, 2H), 3.30-3.17 (m, 4H) , 3.00-2.97 (m, 7H), 2.48-2.44 (m, 3H), 2.39-2.22 (m, 2H), 2.21-2.13 (m, 3H), 2.09-1.88 (m, 2H), 0.97-0.93 (m, 3H), 0.79-0.76 (m, 3H).

[001356] Example of the synthesis of 2-((2S)-1-acryloyl-4-(2-(((2S,4R)-4-(2-(2-(2-(2-((2-(2, 6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl) -5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile (typical compound 518)

[001357] Step 1: Preparation of tert-butyl (2S,4R)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(2-(2-(2-(2-((tetrahydro-2H- pyran-2-yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)pyrrolidine-1-carboxylate

[001358] To a solution of tert-butyl (2S,4R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-hydroxy-pyrrolidine-1-carboxylate (6 g, 18.10 mmol, 1 eq. ) in dry tetrahydrofuran (80 ml) (dried with sodium and distilled), sodium hydrogen (1.45 g, 36.20 mmol, 60% purity, 2 eq.) was added at 20°C. The reaction mixture was stirred at 20°C for 30 minutes. Then a solution of 2-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]ethyl 4-methylbenzenesulfonate (8.3 g, 19.19 mmol, 1.06 eq.) in tetrahydrofuran (40 ml) was added and the reaction mixture was stirred at 50°C for another 14 hours. Ethyl acetate (40 ml) was added and the mixture was washed with saturated aqueous ammonium chloride (40 ml). The organic layer was dried over sodium sulfate and then concentrated in vacuo to obtain a residue. The residue was purified by flash chromatography (silica gel, 0-100%) with ethyl acetate in petroleum ether) to give tert-butyl (2S,4R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-[2- [2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]ethoxy]pyrrolidine-1-carboxylate (3.7 g, 6.25 mmol, 34% yield) as a light yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.67-4.62 (m, 1H), 4.26-4.08 (m, 1H), 3.98 (br s, 1H), 3, 90-3.84 (m, 2H), 3.71-3.65 (m, 11H), 3.65-3.61 (m, 3H), 3.61-3.48 (m, 5H), 3.45-3.37 (m, 1H), 2.19 (td, J=5.4, 12.9 Hz, 1H), 2.09-1.94 (m, 1H), 1.89- 1.69 (m, 2H), 1.62 (br d, J=4.0 Hz, 1H), 1.66-1.60 (m, 1H), 1.58-1.49 (m, 3H ), 1.46 (s, 9H), 1.30-1.24 (m, 1H), 0.88 (s, 9H), 0.07 - -0.03 (m, 6H).

[001359] Step 2: Preparation of tert-butyl (2S,4R)-2-(hydroxymethyl)-4-(2-(2-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy) ethoxy)ethoxy)ethoxy)ethoxy)pyrrolidine-1-carboxylate

[001360] To a solution of tert-butyl (2S,4R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-[2-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy ]ethoxy]ethoxy]pyrrolidine-1-carboxylate (3.7 g, 6.25 mmol, 1 equiv.) in tetrahydrofuran (40 ml) was added tetrabutylammonium fluoride (1 M, 6.25 ml, 1 equiv.) at 20° WITH. The reaction mixture was stirred at 20°C for 6 hours. The reaction mixture was concentrated in vacuo to obtain a residue. The residue was purified by silica gel column chromatography (30% ethyl acetate in petroleum ether to 100% ethyl acetate) to give tert-butyl (2S,4R)-2-(hydroxymethyl)-4-[2-[2-[2-(2- tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]ethoxy]pyrrolidine-1-carboxylate (2.9 g, 6.07 mmol, 97% yield) as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.91 (br d, J=8.4 Hz, 1H), 4.67-4.61 (m, 1H), 4.13-3.97 ( m, 2H), 3.92-3.84 (m, 2H), 3.71-3.62 (m, 13H), 3.61-3.54 (m, 4H), 3.54-3, 48 (m, 1H), 3.41 (br dd, J=4.5, 11.9 Hz, 1H), 2.20-2.11 (m, 1H), 1.89-1.79 (m , 1H), 1.77-1.69 (m, 1H), 1.56 (br s, 1H), 1.64-1.51 (m, 4H), 1.47 (s, 9H).

[001361] Step 3: Preparation of ((2S,4R)-1-methyl-4-(2-(2-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy) ethoxy)ethoxy)pyrrolidin-2-yl)methanol

[001362] To a solution of tert-butyl (2S,4R)-2-(hydroxymethyl)-4-[2-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy] ethoxy] ethoxy] pyrrolidine-1 - carboxylate (9.8 g, 20.52 mmol, 1 eq.) in tetradrum rane (70 ml) was added lithium aluminum hydride (2.34 g, 61.56 mmol, 3 eq.) at 20°C. The reaction mixture was stirred at 60°C for 14 hours. The reaction mixture was cooled to 10°C and tetrahydrofuran (300 ml) was added. Ice water (2 mL) was slowly added to quench the reaction while stirring. Datomite (30 g) was added to the suspension and stirred at 20°C for 10 minutes. The mixture was filtered and the filter cake was washed with tetrahydrofuran (50 ml x 3). The filtrate was concentrated in vacuo to [(2S,4R)-1-methyl-4-[2-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]ethoxy]pyrrolidin-2-yl]methanol ( 6.5 g, 16.60 mmol, 80% yield) as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.68-4.60 (m, 1H), 4.06-3.96 (m, 1H), 3.92-3.83 (m, 2H) , 3.71-3.61 (m, 14H), 3.61-3.54 (m, 2H), 3.54-3.47 (m, 1H), 3.44-3.36 (m, 2H), 2.69-2.59 (m, 1H), 2.39 (dd, J=6.2, 9.7 Hz, 1H), 2.34 (s, 3H), 2.09 (td , J=8.0, 13.4 Hz, 1H), 1.94-1.80 (m, 2H), 1.77-1.68 (m, 1H), 1.66-1.57 (m , 3H), 1.51 (br d, J=3.3 Hz, 1H).

[001363] Step 4: Preparation of tert-butyl 4-((S)-4-((benzyloxy)carbonyl)-3-(cyanomethyl)piperazin-1-yl)-2-(((2S,4R)-1- methyl-4-(2-(2-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)ethoxy)ethoxy)pyrrolidin-2-yl)methoxy)-5.8 -dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate

[001364] To a solution of [(2S,4R)-1-methyl-4-[2-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]ethoxy]pyrrolidin-2-yl]methanol ( 1.37 g, 3.50 mmol, 1.32 eq.), tert-butyl 4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-2-chloro-6.8 -dihydro-5H-pyrido[3,4-d]pyrimidine-7-carboxylate (1.4 g, 2.66 mmol, 1 eq.) in dioxane (35 ml) was added cesium carbonate (2.60 g, 7. 97 mmol, 3 eq.) and methanesulfonato(2-dicyclohexylphosphino-2,6-di-i-propoxy-1,1-biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(ii) ( 222 mg, 0.26 mmol, 0.1 eq.). The reaction mixture was degassed and charged with nitrogen 3 times and then stirred at 90°C for 2 hours. Ethyl acetate (50 ml) and saturated aqueous ammonium chloride (30 ml) were added and the mixture was separated. The aqueous layer was extracted with ethyl acetate (40 ml). The combined organic layer was dried over sodium sulfate and then concentrated in vacuo to leave a residue. The residue was purified by silica gel column chromatography (50% ethyl acetate in petroleum ether to 100% ethyl acetate then 5% methanol in tetrahydrofuran) to give tert-butyl 4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazine- 1-yl]-2-[[(2S,4R)-1-methyl-4-[2-[2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]ethoxy]pyrrolidin-2-yl ]methoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-7-carboxylate (1.6 g, 1.81 mmol, 68% yield) as a light brown gum.

[001365] Step 5: Preparation of benzyl (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy )ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazine-1-carboxylate

[001366] To a solution of tert-butyl 4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-2-[[(2S,4R)-1-methyl-4-[2- [2-[2-(2-tetrahydropyran-2-yloxyethoxy)ethoxy]ethoxy]ethoxy]pyrrolidin-2-yl]methoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-7- carboxylate (1.2 g, 1.36 mmol, 1 eq.) in dichloromethane (15 ml), trifluoroacetic acid (4.62 g, 40.52 mmol, 3.00 ml, 29.78 eq.) was added. The reaction mixture was stirred at 20°C for 2 hours. The reaction solution was poured and added to saturated sodium bicarbonate solution (30 ml) and the mixture was adjusted to pH 7 with triethylamine and then concentrated in vacuo to leave a residue. This product was dissolved in tetrahydrofuran (8 ml) and then stirred with 1 N sodium hydroxide (5 ml) for 10 minutes. Then water (30 ml) and the mixture was extracted with dichloromethane (25 ml x 2). The organic layer was dried over sodium sulfate and then concentrated in vacuo to give the crude product. The crude product was purified by preparative HPLC to give benzyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy ]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (660 mg, 0. 95 mmol, yield 69%) in the form of a colorless gum. LC/MS (ESI) m/z: 698.4 [M+1] ⁺ .

[001367] Step 6: Preparation of benzyl (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy )ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-1 -carboxylate

[001368] Into a mixture of benzyl (2S)-2-(cmanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy ]-1-methyl-pyrrolidin-2-yl]methoxy]-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl]piperazin-1-carboxylate (1.6 g, 2. 29 mmol, 1 equiv.), 1-bromonaphthalene (950 mg, 4.59 mmol, 0.64 ml, 2 equiv.), methanesulfonato(2-dicyclohexylphosphino-2,6-di-i-propoxy-1,1- biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(ii) (192 mg, 0.23 mmol, 0.1 eq.) and cesium carbonate (2.24 g, 6.88 mmol, 3 eq.) dioxane (30 ml) was added. The reaction mixture was degassed and charged with nitrogen three times and then stirred at 90°C for 14 hours. The reaction mixture was concentrated in vacuo to obtain a residue. Saturated aqueous ammonium chloride (40 ml) and water (20 ml) were added and the mixture was extracted with ethyl acetate (40 ml x 2). The organic layer was dried over sodium sulfate and then concentrated in vacuo to obtain a residue. The product was purified by silica gel column chromatography (30-100% ethyl acetate in petroleum ether followed by 5% methanol in tetrahydrofuran) to give benzyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)- 4-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro- 5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (1.05 g, 1.03 mmol, 45% yield, 81% purity) as a light brown gum. LC/MS (ESI) m/z: 824.3 [M+1] ⁺ .

[001369] Step 7: Preparation of tert-butyl (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-(2-hydroxyethoxy)ethoxy )ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazine -1-carboxylate

[001370] To a solution of benzyl (2S)-2-(cmanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy ]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (1.17 g, 1.42 mmol, 1 eq.), ammonium hydroxide (1.82 g, 17.14 mmol, 2 mL, 33% purity, 12.07 eq.) in methanol (30 mL) added palladium on coal (150 mg, purity 5%). The reaction mixture was degassed and charged with hydrogen three times and then stirred at 20°C with [0.1 MPa (15 psi)] for 4 hours. The reaction mixture was then stirred at 20°C with [0.1 MPa (15 psi)] for an additional 2 hours. The reaction mixture was filtered and the filtrate was concentrated in vacuo to give 2-[(2S)-4-[2-[[ (2S,4R)-4-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)- 6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-2-yl]acetonitrile (980 mg) as a colorless gum. To a solution of 2-[(2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidine -2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-2-yl]acetonitrile (980 mg, 1 .42 mmol, 1 eq.) in dichloromethane (20 ml) was added triethylamine (431 mg, 4.26 mmol, 3 eq.) and di-tert-butyl dicarbonate (1.55 g, 7.10 mmol, 1. 63 ml, 5 eq.). The reaction mixture was stirred at 15°C for 14 hours. The reaction mixture was filtered and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified with silica gel (0-5% methanol in dichloromethane) to give tert-butyl (2S)-2-(tzmanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2 -(2-hydroxyethoxy)ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d] pyrimidin-4-yl]piperazine-1-carboxylate (1 g) as a colorless gum. LC/MS (ESI) m/z: 690.3 [M+1] ⁺ .

[001371] Step 8: Preparation of tert-butyl (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-1-methyl-4-(2-(2-(2-(2 -(tosyloxy)ethoxy)ethoxy)ethoxy)ethoxy)pyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4 -yl)piperazine-1-carboxylate

[001372] To a solution of tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy ]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1 -carboxylate (500 mg, 0.63 mmol, 1 eq.) in dichloromethane (8 ml) was added 4-methylbenzene-1-sulfonyl chloride (241 mg, 1.27 mmol, 2 eq.), triethylamine (192 mg, 1. 90 mmol, 3 eq.) and N,N-dimethylpyridin-4-amine (15 mg, 0.13 mmol, 0.2 eq.). The reaction mixture was stirred at 25°C for 14 hours. The reaction mixture was filtered and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by preparative TLC (10% methanol in dichloromethane) to give tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-1-methyl-4-[2-[2 -[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]ethoxy]pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4 -d]pyrimidin-4-yl]piperazin-1-carboxylate (400 mg, 0.42 mmol, 66% yield) as a colorless gum. LC/MS (ESI) m/z: 944.3 [M+1] ⁺ .

[001373] Step 9: Preparation of tert-butyl (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-(2-(1, 3-dioxoisoindolin-2-yl)ethoxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3, 4-d]pyrimidin-4-yl)piperazine-1-carboxylate

[001374] A mixture of tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-1-methyl-4-[2-[2-[2-[2-(polysulfoxide )ethoxy]ethoxy]ethoxy]ethoxy]pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin- 1-carboxylate (200 mg, 0.21 mmol, 1 eq.) and (1,3-dioxoisoindolin-2-yl)potassium (78 mg, 0.42 mmol, 2 eq.) in N,N-dimethylformamide (4 ml) was stirred at 80°C for 2 hours. Ethyl acetate (40 ml) was added and the mixture was washed with water (30 ml). The organic layer was dried over sodium sulfate and then concentrated in vacuo to obtain a residue. The residue was purified by preparative TLC (10% methanol in dichloromethane) to give tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2- [2-(1,3-dioxo-3a,7a-dihydroisoindol-2-yl)ethoxy]ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)- 6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (140 mg, 0.15 mmol, 72% yield) as a colorless gum. LC/MS (ESI) m/z: 919.4 [M+1] ⁺ .

[001375] Step 10: Preparation of tert-butyl (S)-4-(2-(((2S,4R)-4-(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethoxy)- 1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine -1-carboxylate

[001376] To a mixture of tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-(1,3- dioxo-3a,7a-dihydroisoindol-2-yl)ethoxy]ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H- pyrido[3,4-d]pyrimidin-4-yl]piperazin-1-carboxylate (140 mg, 0.15 mmol, 1 eq.) in ethanol (2 ml) was added hydrazine hydrate (155 mg, 3.04 mmol, 0 .2 ml, purity 98%, 20 eq.). The reaction mixture was stirred at 70°C for 5 hours. The reaction mixture was filtered and the solid was washed with ethanol (30 ml). The filtrate was concentrated in vacuo to give tert-butyl (2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethoxy]-1 -methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]-2-(cyanomethyl)piperazine- 1-carboxylate (140 mg) as a colorless gum.

[001377] Step 11: Preparation of tert-butyl (2S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-(2-((2 -(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene- 1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazine-1-carboxylate

[001378] Tert-Butyl (2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethoxy]-1-methyl mixture -pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]-2-(cyanomethyl)piperazine-1- carboxylate (70 mg, 0.088 mmol, 1 eq.), 2-(2,6-dioxo-3-piperidyl)-4-fluoro-isoindoline-1,3-dione (49 mg, 0.18 mmol, 2 eq. ) and N,N-diisopropylethylamine (46 mg, 0.35 mmol, 4 eq.) in dimethyl sulfoxide (2 ml) were heated at 90°C for 3 hours. The reaction mixture was poured into water (25 ml) and then extracted with ethyl acetate (20 ml). The organic layer was washed with brine (20 ml), dried over sodium sulfate and then concentrated in vacuo to leave a residue. The residue was purified by preparative TLC (10% methanol in dichloromethane) to give tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2- [2-[[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2- yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (25 mg, 0.021 mmol, 24% yield , purity 88%) was obtained as a yellow solid. LC/MS (ESI) m/z: 1045.6 [M+1] ⁺ .

[001379] Step 12: Preparation of 2-((2S)-4-(2-(((2S,4R)-4-(2-(2-(2-(2-((2-(2,4- dioxocyclohexyl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7 ,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[001380] To a solution of tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-[[2-( 2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-( 1-Naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (30 mg, 0.028 mmol, 1 eq.) in dichloromethane (5 ml) was added trifluoroacetic acid (1.54 g, 13.51 mmol, 1 ml, 470.55 eq.). The mixture was stirred at 20°C for 1 hour. The reaction mixture was concentrated in vacuo to give the compound 2-[(2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-[[2-(2,6 -dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl )-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-2-yl]acetonitrile (30 mg, 0.028 mmol, 99% yield, trifluoroacetate) as a yellow oil.

[001381] Step 13: Preparation of 2-((2S)-1-acryloyl-4-(2-(((2S,4R)-4-(2-(2-(2-(2-((2-( 2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1- yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[001382] To a mixture of 2-[(2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-[[2-(2,6-dioxo- 3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6 ,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-2-yl]acetonitrile (30 mg, 0.028 mmol, 1 eq., trifluoroacetate) in dichloromethane (5 ml) was added 6-lutidine (30 mg, 0.28 mmol, 10 eq.), then prop-2-enoyl chloride (2 mg, 0.025 mmol, 0.9 eq.) in dichloromethane (2 ml) was added at minus 65°C, then the reaction mixture was stirred at minus 65°C for 10 minutes. Water (10 ml) was added and stirred for 0.5 minutes. The aqueous phase was extracted with dichloromethane (20 ml x 2) and concentrated in vacuo. The residue was purified by semipreparative reverse phase HPLC. Compound 2-[(2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-[[2-(2,6-dioxo-3-piperidyl) -1,3-dioxo-isoindolin-4-yl]amino]ethoxy]ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro -5H-pyrido[3,4-d]pyrimidin-4-yl]-1-prop-2-enoyl-piperazin-2-yl]acetonitrile (15.8 mg, 0.015 mmol, 54% yield, 97% purity) was obtained as a yellow solid. LC/MS (ESI) m/z: 999.5 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.09 (s, 1H), 8.22-8.16 (m, 1H), 7.97-7.90 (m, 1H), 7 .65 (d, J=8.3 Hz, 1H), 7.61-7.51 (m, 3H), 7.47 (t, J=7.8 Hz, 1H), 7.23 (d, J=7.3 Hz, 1H), 7.14 (d,J=8.7 Hz, 1H), 7.04 (d, J=7.0 Hz, 1H), 6.88 (br s, 1H ), 6.60 (br t, J=5.7 Hz, 1H), 6.20 (dd, J=2.0, 16.6 Hz, 1H), 5.85-5.75 (m, 1H ), 5.05 (dd, J=5.4, 12.9 Hz, 1H), 5.02-4.73 (m, 1H), 4.26 (br dd, J=4.6, 10, 9 Hz, 1H), 4.14 (s, 2H), 4.12-4.07 (m, 1H), 4.06-3.94 (m, 3H), 3.62 (br t, J= 5.4 Hz, 2H), 3.57-3.53 (m, 4H), 3.52-3.49 (m, 4H), 3.49-3.47 (m, 2H), 3.46 (br s, 2H), 3.43 (br d, J=6.1 Hz, 2H), 3.27 (br dd, J=6.2, 9.5 Hz, 3H), 3.21-3 .17 (m, 2H), 3.10-2.83 (m, 4H), 2.75-2.66 (m, 2H), 2.60 (br d, J=2.7 Hz, 4H) , 2.35-2.31 (m, 3H), 2.17 (br dd, J=6.0, 9.5 Hz, 1H), 2.07-1.98 (m, 1H), 1, 91-1.78 (m, 2H).

[001383] Example of the synthesis of 2-((2S)-1-acryloyl-4-(2-(((2S,4R)-4-(2-(2-(2-((2-(2,6-dioxosheridine -3-yl)-1-oxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7, 8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile (typical compound 521)

[001384] Step 1: Preparation of methyl 4-((tert-butyldimethylsilyl)oxy)-2-methylbenzoate

[001385] To a solution of methyl 4-hydroxy-2-methyl benzoate (5.00 g, 30.09 mmol, 1.00 eq.) in dimethylformamide (100 ml) was added imidazole (6.15 g, 90.27 mmol , 3.00 eq.). The mixture was stirred at 15°C for 0.5 h and then chlorotrimethylsilane (6.80 g, 45.14 mmol, 5.5 mL, 1.50 eq.) was added. The resulting mixture was stirred at 15°C for another 14.5 hours. The mixture was poured into saturated brine (300 ml) and then extracted with ethyl acetate (300 ml * 2). The combined organic layers were washed with 1 M hydrochloric acid (300 ml * 2), brine (300 ml * 3), dried over sodium sulfate, filtered and concentrated in vacuo to give methyl 4-[tert-butyl(dimethyl)silyl]oxy-2 -methyl benzoate (8.00 g, 28.53 mmol, 95% yield) as a colorless oil. LC/MS (ESI) m/z: 281.2 [M+1] ⁺ .

[001386] Step 2: Preparation of methyl 2-(bromomethyl)-4-((tert-butyldimethylsilyl)oxy)benzoate

[001387] To a solution of methyl 4-[tert-butyl(dimethyl)silyl]oxy-2-methyl-benzoate (3.00 g, 10.70 mmol, 1.00 eq.) in carbon tetrachloride (40 ml) was added N -bromosuccinimide (2.29 g, 12.84 mmol, 1.20 eq.) and 2,2-azobisobutyronitrile (88 mg, 0.54 mmol, 0.05 eq.). The mixture was stirred at 15°C for 0.5 hours, then heated to 80°C and stirred at 80°C for another 2.5 hours. The mixture was poured into water (300 ml), the organic layer was separated. The aqueous layer was extracted with dichloromethane (200 ml * 2). The combined organic layers were washed with saturated brine (300 ml * 3), dried over sodium sulfate, filtered and concentrated in vacuo to give methyl 2-(bromomethyl)-4-[tert-butyl(dimethyl)silyl]oxy-benzoate (4.00 g, 8.39 mmol, yield 78%, purity 75%) as a light yellow oil. LC/MS (ESI) m/z: 359.0/361.0 [M+1] ⁺ .

[001388] Step 3: Preparation of tert-butyl 5-amino-4-(5-((tert-butyldimethylsilyl)oxy)-1-oxoisoindolin-2-yl)-5-oxopentanoate

[001389] To a mixture of methyl 2-(bromomethyl)-4-[tert-butyl(dimethyl)silyl]oxy-benzoate (9.12 g, 25.38 mmol, 1.00 eq.) and diisopropylethylamine (13.12 g , 101.52 mmol, 18 ml, 4.00 eq.) in acetonitrile (70 ml) was added tert-butyl 4,5-diamino-5-oxo-pentanoate (5.13 g, 25.38 mmol, 1.00 eq.) in one portion at 0°C in a nitrogen atmosphere. The mixture was stirred at a temperature of 60-80°C for 16 hours. The mixture was concentrated in vacuo. Then water (50 ml) was added. The aqueous phase was extracted with ethyl acetate (100 ml * 3). The combined organic phase was washed with brine (100 ml * 2), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (petroleum ether/ethyl acetate 3/1 ~ 1/1, then dichloromethane/methanol 10/1). Product tert-butyl 5-amino-4-[5-[tert-butyl (dimethyl)silyl]oxy-1-oxo-isoindolin-2-yl]-5-oxo-pentanoate (3.12 g, 6.68 mmol , yield 26%, purity 96%) was obtained as a colorless oil. LC/MS (ESI) m/z: 449.1 [M+1] ⁺ .

[001390] Step 4: Preparation of tert-butyl 5-amino-4-(5-hydroxy-1-oxoisoindolin-2-yl)-5-oxopentanoate

[001391] To a solution of tert-butyl 5-amino-4-[5-[tert-butyl(dimethyl)silyl]oxy-1-oxo-isoindolin-2-yl]-5-oxo-pentanoate (3.12 g, 6.95 mmol, 1.00 eq.) in methanol (30 ml) was added tetrabutylammonium fluoride (1 M in tetrahydrofuran, 7 ml, 1.00 eq.) in tetrahydrofuran (18 ml). The reaction mixture was stirred at 25°C for 16 hours. The mixture was concentrated in vacuo. Water (100 ml) was added and the aqueous phase was extracted with dichloromethane (100 ml *2). The combined organic phase was washed with brine (100 ml * 3), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (0-70% ethyl acetate in petroleum ether). Product tert-butyl 5-amino-4-(5-hydroxy-1-oxo-isoindolin-2-yl)-5-oxo-pentanoate (2.50 g, 6.78 mmol, 97% yield, 90% purity) was obtained as a light yellow solid. LC/MS (ESI) m/z: 357.0 [M+23] ⁺ .

[001392] Step 5: Preparation of tert-butyl (2S)-4-(2-(((2S,4R)-4-(2-(2-(2-((2-(1-amino-5-( tert-butoxy)-1,5-dioxopentan-2-yl)-1-oxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1 -yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[001393] To a solution of tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-1-methyl-4-[2-[2-[2-(p-tolylsulfonyloxy )ethoxy]ethoxy]ethoxy]pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-1- carboxylate (140 mg, 0.16 mmol, 1 eq.) and tert-butyl 5-amino-4-(5-hydroxy-1-oxo-isoindolin-2-yl)-5-oxo-pentanoate (78 mg, 0 .23 mmol, 1.5 eq.) in acetonitrile (3 ml), potassium carbonate (64 mg, 0.47 mmol, 3 eq.) was added and the mixture was stirred at 90°C for 12 hours. The mixture was filtered and the filtrate was concentrated to give the crude product, which was purified by preparative TLC (10:1 dichloromethane:methanol) to give tert-butyl(2S)-4-[2-[[(2S,4R)-4-[2 -[2-[2-[2-(4-tert-butoxy-1-carbamoyl-4-oxo-butyl)-1-oxo-isoindolin-5-yl]oxyethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidine -2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate ( 110 mg, 103.55 µmol, 66.6% yield) as a brown oil.

[001394] Step 6: Preparation of 2-((2S)-4-(2-(((2S,4R)-4-(2-(2-(2-((2-(2,6-dioxopiperidine-3 -yl)-1-oxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8- tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[001395] To a mixture of tert-butyl (2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-(4-tert-butoxy-1-carbamoyl- 4-oxo-butyl)-1-oxo-isoindolin-5-yl]oxyethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro -5H-pyrido[3,4-d]pyrimidin-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (110 mg, 0.1 mmol, 1 eq.) in acetonitrile (6 ml) added benzenesulfonic acid (66 mg, 0.4 mmol, 4 eq.) in one portion at 25°C under nitrogen, the mixture was stirred at 80°C for 10 hours. The mixture was concentrated in vacuo. The residue was purified by preparative HPLC to give 2-[(2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-(2,6-dioxo-3-piperidyl )-1-oxo-isoindolin-5-yl]oxyethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[ 3,4-d]pyrimidin-4-yl]piperazin-2-yl]acetonitrile (45 mg, 0.05 mmol, 48% yield) as a yellow oil.

[001396] Step 7: Preparation of 2-((2S)-1-acryloyl-4-(2-(((2S,4R)-4-(2-(2-(2-((2-(2,6 -dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)oxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6, 7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[001397] To a mixture of 2-[(2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-(2,6-dioxoo-3-piperidyl) -1-oxo-isoindolin-5-yl]oxyethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3 ,4-d]pyrimidin-4-yl]piperazin-2-yl]acetonitrile (45 mg, 0.05 mmol, 1 eq.) in dichloromethane (10 ml) was added 2,6-lutidine (54 mg, 0.51 mmol, 10 eq.) in one portion at minus 78°C under nitrogen, then prop-2-enoyl chloride (4.1 mg, 0.05 mmol, 0.03 mL, 0.9 eq.) in dichloromethane (1 ml). The mixture was stirred at minus 78°C for 15 minutes. The mixture was quenched with water (20 ml). The aqueous phase was extracted with ethyl acetate (15 ml x 3). The combined organic phase was washed with brine (15 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative HPLC to give 2-[(2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[2-(2,6-dioxo-3-piperidyl )-1-oxo-isoindolin-5-yl]oxyethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[ 3,4-d]pyrimidin-4-yl]-1-prop-2-enoyl-piperazin-2-yl]acetonitrile (13 mg, 0.012 mmol, 25% yield, 98% purity, formate) as a white solid . LC/MS (ESI) m/z: 471.8 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.96 (s, 1H), 8.24 (s, 1H), 8.20-8.14 (m, 1H), 7.95-7 .89 (m, 1H), 7.66-7.58 (m, 2H), 7.56-7.51 (m, 2H), 7.45-7.3 (m, 1H), 7.25 -7.2 (m, 1H), 7.18-7.1 (m, 1H), 7.06-6.9 (m, 1H), 6.88-6.5 (m, 1H), 6 ,23-6.14 (m, 1H), 5.8-5.5 (m, 1H), 5.06-5.0 (m, 1H), 4.98-4.6 (m, 1H) , 4.49-4.32 (m, 2H), 4.27-4.21 (m, 2H), 4.19-4.15 (m, 2H), 4.15-3.93 (m, 8H), 3.79-3.74 (m, 2H), 3.61-3.57 (m, 3H), 3.55-3.53 (m, 3H), 3.51-3.49 ( m, 4H), 3.30-3.14 (m, 2H), 3.04-2.84 (m, 4H), 2.75-2.65 (m, 2H), 2.39-2, 33 (m, 5H), 2.16-2.1 (m, 1H), 2.00-1.92 (m, 1H), 1.88-1.79 (m, 2H).

[001398] Example of synthesis of (S)-N-((S)-2-((S)-2-(4-(4-(2-(2-(2-(((3R,5S)-5- (((4-((S)-4-acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4- d]pyrimidin-2-yl)oxy)methyl)-1-methylpyrrolidin-3-yl)oxy)ethoxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)-1- cyclohexyl-2-oxoethyl)-2-(methylamino)propanamide (representative compound 548)

[001399] Step 1: Preparation of benzyl (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-1-methyl-4-(2-(2-(2-(tosyloxy)ethoxy )ethoxy)ethoxy)pyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperazin-1- carboxylate

[001400] To a mixture of benzyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]-1-methyl -pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-e]pyrimidin-4-yl]piperazine-1-carboxylate (160 mg, 0 .20 mmol, 1 eq.) in dichloromethane (2 ml) was added triethylamine (0.1 ml, 3 eq.), p-toluenesulfonyl chloride (78 mg, 0.41 mmol, 2 eq.) and dimethylaminopyridine (25 mg, 0 .20 mmol, 1 eq.). The reaction mixture was then stirred at 15°C for 12 hours. The reaction mixture was filtered. The filtrate was purified by preparative TLC (dichloromethane:methanol 10:1) to obtain the product. The compound benzyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-1-methyl-4-[2-[2-[2-(p-tolylsulfonyloxyoxy)ethoxy]ethoxy]ethoxy ]pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (160 mg, 0 .17 mmol, 83.5% yield) was obtained as a yellow solid. LC/MS (ESI) m/z: 934.5 [M+1] ⁺ .

[001401] Step 2: Preparation of benzyl (S)-4-(2-(((2S,4R)-4-(2-(2-(2-((4-(2-((S)-1- ((S)-2-((S)-2-((tert-butoxycarbonyl)(methyl)amino)propanamido)-2-cyclohexylacetyl)pyrrolidin-2-yl)thiazol-4-yl)naphthalene-1-yl) oxy)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4- yl)-2-(cyanomethyl)piperazine-1-carboxylate

[001402] To a mixture of tert-butyl N-[(1S)-2-[[(1S)-1-cyclohexyl-2-[(2S)-2-[4-(4-hydroxy-1-naphthyl)thiazol- 2-yl]pyrrolidin-1-yl]-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]-1-methyl-carbamate (106 mg, 0.17 mmol, 1 eq.) and benzyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-1-methyl-4-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy] pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (160 mg, 0. 17 mmol, 1 eq.) in acetonitrile (5 ml) was added potassium carbonate (47 mg, 0.34 mmol, 2 eq.). The mixture was stirred at 85°C for 12 hours. The mixture was quenched with water (20 ml). The mixture was then extracted with dichloromethane (30 ml x 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to leave a residue. The residue was purified by preparative TLC (dichloromethane:methanol 10:1). Benzyl (2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[[4-[2-[(2S)-1-[(2S)-2- [[(2S)-2-[tert-butoxycarbonyl(methyl)amino]propanoyl]amino]-2-cyclohexyl-acetyl]pyrrolidin-2-yl]thiazol-4-yl]-1-naphthyl]oxy]ethoxy]ethoxy ]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]-2- (cyanomethyl)piperazine-1-carboxylate (105 mg, 0.07 mmol, 44.3% yield) was obtained as a yellow solid. LC/MS (ESI) m/z: 1383.6 [M+1] ⁺ .

[001403] Step 3: Preparation of tert-butyl ((S)-1-(((S)-2-((S)-2-(4-(4-(2-(2-(2-((( 3R,5S)-5-(((4-((S)-3-(cyanomethyl)piperazin-1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3 ,4-d]pyrimidin-2-yl)oxy)methyl)-1-methylpyrrolidin-3-yl)oxy)ethoxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl) -1-cyclohexyl-2-oxoethyl)amino)-1-oxopropane-2-l)(methyl)carbamate

[001404] To a mixture of benzyl (2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[[4-[2-[(2S)-1-[( 2S)-2-[[(2S)-2-[tert-butoxycarbonyl(methyl)amino]propanoyl]amino]-2-cyclohexyl-acetyl]pyrrolidin-2-yl]thiazol-4-yl]-1-naphthyl] oxy]ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-e]pyrimidine-4- yl]-2-(cyanomethyl)piperazine-1-carboxylate (105 mg, 0.07 mmol, 1 eq.) and ammonium hydroxide (0.2 ml) in tetrahydrofuran (0.5 ml) and methanol (5 ml) were added catalyst palladium on activated carbon (30 mg, purity 10%). The mixture was degassed and charged with hydrogen three times. The mixture was then stirred at 25°C for 12 hours. The mixture was filtered. The filtrate was purified by preparative HPLC. Tert-butyl compound N-[(1S)-2-[[(1S)-2-[(2S)-2-[4-[4-[2-[2-[2-[(3R,5S)- 5-[[4-[(3S)-3-(cyanomethyl)piperazin-1-yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2 -yl]oxymethyl]-1-methyl-pyrrolidin-3-yl]oxyethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-1-cyclohexyl-2-oxo-ethyl] amino]-1-methyl-2-oxo-ethyl]-N-methyl-carbamate (35 mg, 0.03 mmol, 36.9% yield) was obtained as a white solid. LC/MS (ESI) m/z: 1248.8 [M+1] ⁺ .

[001405] Step 4: Preparation of tert-butyl ((S)-1-(((S)-2-((S)-2-(4-(4-(2-(2-(2-((( 3R,5S)-5-(((4-((S)-4-acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(naphthalene-1-yl)-5,6,7,8 -tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)-1-methylpyrrolidin-3-yl)oxy)ethoxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin- 1-yl)-1-cyclohexyl-2-oxoethyl)amino)-1-oxopropan-2-yl)(methyl)carbamate

[001406] To a mixture of tert-butyl N-[(1S)-2-[[(1S)-2-[(2S)-2-[4-[4-[2-[2-[2-[(3R ,5S)-5-[[4-[(3S)-3-(cyanomethyl)piperazin-1-yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d ]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl]oxyethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-1-cyclohexyl-2- oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]-N-methyl-carbamate (35 mg, 0.03 mmol, 1 eq.) in dichloromethane (5 ml) was added 2,6-lutidine (30 mg, 0.28 mmol, 10 eq.) and prop-2-enoyl chloride (2 mg, 0.9 eq.) at minus 78°C. The mixture was then stirred at minus 78°C for 30 minutes. The mixture was diluted with water (3 ml). The mixture was then extracted with ethyl acetate (30 ml x 3). The combined organic layers were washed with water (30 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to leave a residue. Tert-butyl N-[(1S)-2-[[(1S)-2-[(2S)-2-[4-[4-[2-[2-[2-[(3R,5S)-5 -[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoyl-piperazin-1-yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3 ,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl]oxyethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-1- Cyclohexyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]-N-methyl-carbamate (35 mg) was obtained as a pink oil. LC/MS (ESI) m/z: 1302.9 [M+1] ⁺ .

[001407] Step 5: Preparation of (S)-N-((S)-2-((S)-2-(4-(4-(2-(2-(2-(((3R,5S)- 5-(((4-((S)-4-acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3, 4-d]pyrimidin-2-yl)oxy)methyl)-1-methylpyrrolidin-3-yl)oxy)ethoxy)ethoxy)ethoxy)naphthalene-1-yl)thiazol-2-yl)pyrrolidin-1-yl)- 1-cyclohexyl-2-oxoethyl)-2-(methylamino)propanamide

[001408] To a mixture of tert-butyl N-[(1S)-2-[[(1S)-2-[(2S)-2-[4-[4-[2-[2-[2-[(3R ,5S)-5-[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoyl-piperzin-1-yl]-7-(1-naphthyl)-6,8-dihydro- 5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl]oxyethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1- yl]-1-cyclohexyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]-1-methyl-carbamate (35 mg, 0.03 mmol, 1 eq.) in dichloromethane (2 ml ) trifluoroacetic acid (0.2 ml) was added. The mixture was stirred at 25°C for 30 minutes. The mixture was added dropwise to the saturated sodium bicarbonate solution. The mixture was then extracted with a solution (dichloromethane:methanol 10:1, 30 ml x 2). The combined organic layers were concentrated in vacuo to leave a residue. The residue was purified by preparative HPLC. Compound (2S)-N-[(1S)-2-[(2S)-2-[4-[4-[2-[2-[2-[(3R,5S)-5-[[4-[ (3S)-3-(cyanomethyl)-4-prop-2-enoyl-piperazin-1-yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine -2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl]oxyethoxy]ethoxy]ethoxy]-1-naphthyl]thiazol-2-yl]pyrrolidin-1-yl]-1-cyclohexyl-2-oxo- ethyl]-2-(methylamino)propanamide (9.4 mg, 0.007 mmol, 26.3% yield, 98.7% purity, trifluoroacetates) was obtained as a white solid. LC/MS (ESI) m/z: 1203.6 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.13-9.85 (m, 1H), 8.83-8.70 (m, 2H), 8.28-8.22 (m, 1H), 8.20-8.13 (m, 1H), 7.97-7.90 (m, 1H), 7.73-7.63 (m, 2H), 7.61-7.50 ( m, 5H), 7.49-7.43 (m, 1H), 7.21-7.19 (d, J = 7.46 Hz, 1H), 7.03-7.01 (d, J = 8.0 Hz, 1H), 6.94-6.73 (m, 1H), 6.25-6.12 (m, 1H), 5.82-5.72 (m, 1H), 5.47 -5.39 (m, 1H), 5.06-4.91 (m, 1H), 4.81-4.70 (m, 1H), 4.65-4.38 (m, 5H), 4 ,34-4.29 (m, 2H), 4.27-4.22 (m, 1H), 4.14 (s, 2H), 4.10-4.00 (m, 3H), 3.94 -3.67 (m, 12H), 3.63-3.54 (m, 9H), 3.24-3.15 (m, 2H), 3.01-2.93 (m, 5H), 2 .89 (s, 1H), 2.73 (s, 1H), 2.28-2.17 (m, 2H), 2.08-1.96 (m, 3H), 1.79-1.64 (m, 4H), 1.62-1.53 (m, 2H), 1.35-1.33 (d, J=6.85 Hz, 3H), 1.24-1.00 (m, 6H ).

[001409] Example of synthesis of (2R,3S,4R,5S)-N-(4-((2-(2-(2-(((3R,5S)-5-(((4-((S)- 4-acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy) methyl)-1-methylpyrrolidin-3-yl)oxy)ethoxy)ethoxy)ethyl)carbamoyl)-2-methoxyphenyl)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl) -4-cyano-5-neopentylpyrrolidine-2-carboxamide (representative compound 552)

[001410] Step 1: Preparation of tert-butyl (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-(2-(2-(2-(1,3-dioxoisoindoline -2-yl)ethoxy)ethoxy)ethoxy)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine -4-yl)piperazine-1-carboxylate

[001411] To a mixture of tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-1-methyl-4-[2-[2-[2-(p-tolylsulfonyloxy )ethoxy]ethoxy]ethoxy]pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazin-1- carboxylate (300 mg, 0.33 mmol, 1 eq.) in N,N-dimethylformamide (4 ml) was added (1,3-dioxoisoindolin-2-yl)potassium (123 mg, 0.67 mmol, 2 eq.) , then the reaction mixture was stirred at 80°C for 2 hours. Water (25 ml) was added and the aqueous phase was extracted with ethyl acetate (25 ml x 3). The combined organic phase was washed with brine (25 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (1:0 to 30:1 dichloromethane:methanol) to give tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4 -[2-[2-[2-(1,3-dioxoisoindolin-2-yl)ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6 ,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]piperazine-1-carboxylate (220 mg, 0.25 mmol, 75% yield) as a yellow solid. LC/MS (ESI) m/z: 875.5 [M+1] ⁺ .

[001412] Step 2: Preparation of tert-butyl (S)-4-(2-(((2S,4R)-4-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)-1-methylpyrrolidine-2 -yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[001413] To a solution of tert-butyl (2S)-2-(cyanomethyl)-4-[2-[[(2S,4R)-4-[2-[2-[2-(1,3-dioxoisoindoline-2 -yl)ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4 -yl]piperazine-1-carboxylate (150 mg, 0.17 mmol, 1 eq.) in ethanol (5 ml) was added hydrazine hydrate (86 mg, 1.71 mmol, 10 eq.). The mixture was stirred at 80°C for 2 hours. The reaction mixture was cooled to 20°C, then ethyl acetate (20 ml) was added, the mixture was filtered and concentrated in vacuo. Compound tert-butyl (2S)-4-[2-[[(2S,4R)-4-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy ]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (141 mg) was obtained as yellow gum. LC/MS (ESI) m/z: 745.5 [M+1] ⁺ .

[001414] Step 3: Preparation of tert-butyl (S)-4-(2-(((2S,4R)-4-(2-(2-(2-(4-((2R,3S,4R,5S )-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamido)-3-methoxybenzamido)ethoxy)ethoxy)ethoxy)- 1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine -1-carboxylate

[001415] To a mixture of tert-butyl (2S)-4-[2-[[(2S,4R)-4-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]-1-methyl-pyrrolidine-2 -yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (40 mg , 0.054 mmol, 1.1 eq.) and 4-[[(2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluoro- phenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3-methoxy-benzoic acid (30 mg, 0.049 mmol, 1 eq.) in N,N-dimethylformamide (1 ml) 1-hydroxybenzotriazole (10 mg, 0.073 mmol, 1.5 eq.), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (14 mg, 0.073 mmol, 1.5 eq.), and N were added. N-diisopropylethylamine (19 mg, 0.15 mmol, 3 eq.), the reaction mixture was stirred at 25°C for 12 hours. Water (0.1 ml) was added, then the mixture was concentrated in vacuo. The reaction mixture was purified by preparative TLC (dichloromethane:methanol 15:1). Tert-butyl compound (2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[[4-[[(2R,3S,4R,5S)-3- (3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino]-3- methoxy-benzoyl]amino]ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d] Pyrimidin-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (30 mg, 0.022 mmol, 46% yield) was obtained as a yellow oil. LC/MS (ESI) m/z: 1343.6 [M+1] ⁺ .

[001416] Step 4: Preparation of (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-N-(4- ((2-(2-(2-(((3R,5S)-5-(((4-((S)-3-(cyanomethyl)piperazin-1-yl)-7-(naphthalene-1-yl )-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)-1-methylpyrrolidin-3-yl)oxy)ethoxy)ethoxy)ethyl)carbamoyl)-2- methoxyphenyl)-5-neopentylpyrrolidine-2-carboxamide

[001417] To a mixture of tert-butyl (2S)-4-[2-[[(2S,4R)-4-[2-[2-[2-[[4-[[(2R,3S,4R,5S )-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-(2,2-dimethylpropyl)pyrrolidine-2-carbonyl]amino ]-3-methoxy-benzoyl]amino]ethoxy]ethoxy]ethoxy]-1-methyl-pyrrolidin-2-yl]methoxy]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3, 4-d]pyrimidin-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (30 mg, 0.022 mmol, 1 eq.) was added to dichloromethane (5 ml) with trifluoroacetic acid (1.54 g, 13.51 mmol, 1 ml, 604.79 eq.), then the reaction mixture was stirred at 25°C for 30 minutes. The reaction mixture was concentrated in vacuo. Compound (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-N-[4-[2 -[2-[2-[(3R,5S)-5-[[4-[(3S)-3-(cyanomethyl)piperazin-1-yl]-7-(1-naphthyl)-6,8-dihydro -5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl]oxyethoxy]ethoxy]ethylcarbamoyl]-2-methoxy-phenyl]-5-(2,2 -Dimethylpropyl)pyrrolidine-2-carboxamide (30 mg, 0.022 mmol, 99% yield, trifluoroacetate) was obtained as a yellow oil.

[001418] Step 5: Preparation of (2R,3S,4R,5S)-N-(4-((2-(2-(2-(((3R,5S)-5-(((4-((S )-4-acryloyl-3-(cyanomethyl)piperazin-1-yl)-7-(naphthalene-1-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl) oxy)methyl)-1-methylpyrrolidin-3-yl)oxy)ethoxy)ethoxy)ethyl)carbamoyl)-2-methoxyphenyl)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2- fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamide

[001419] To a mixture of (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-N-[ 4-[2-[2-[2-[(3R,5S)-5-[[4-[(3S)-3-(cyanomethyl)piperazin-1-yl]-7-(1-naphthyl)-6 ,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl]oxyethoxy]ethoxy]ethylcarbamoyl]-2-methoxy-phenyl]-5- (2,2-dimethylpropyl)pyrrolidine-2-carboxamide (30 mg, 0.022 mmol, 1 eq., trifluoroacetate) in dichloromethane (5 ml) was added 2,6-lutidine (24 mg, 0.22 mmol, 10 eq.) , then prop-2-enoyl chloride (2 mg, 0.020 mmol, 0.9 eq.) was added at −65°C for 10 minutes. Water (10 ml) was added and stirred for 0.5 minutes. The aqueous phase was extracted with dichloromethane (20 ml x 2), and concentrated in vacuo. The residue was purified by semipreparative reverse phase HPLC. Compound (2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-N-[4-[2 -[2-[2-[(3R,5S)-5-[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoyl-piperazin-1-yl]-7-(1 -naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-2-yl]oxymethyl]-1-methyl-pyrrolidin-3-yl]oxyethoxy]ethoxy]ethylcarbamoyl]-2-methoxy- Phenyl]-5-(2,2-dimethylpropyl)pyrrolidine-2-carboxamide (6.0 mg, 0.004 mmol, 20% yield, 99% purity, formate) was obtained as a white solid. LC/MS (ESI) m/z: 640.0 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.41 (s, 1H), 8.49 (br t, J=5.5 Hz, 1H), 8.31 (d, J=8, 4 Hz, 1H), 8.22 (s, 1H), 8.20-8.14 (m, 1H), 7.96-7.89 (m, 1H), 7.72 (br t, J= 6.8 Hz, 1H), 7.64 (d, J=8.3 Hz, 1H), 7.60-7.50 (m, 5H), 7.50-7.43 (m, 2H), 7.41-7.32 (m, 3H), 7.21 (d, J=7.3 Hz, 1H), 6.98-6.74 (m, 1H), 6.25-6.11 ( m, 1H), 5.78 (br d, J=12.5 Hz, 1H), 5.06-4.72 (m, 1H), 4.62-4.51 (m, 2H), 4, 51-4.28 (m, 2H), 4.24 (br dd, J=4.7, 10.8 Hz, 1H), 4.15-3.93 (m, 8H), 3.91 (s , 3H), 3.55-3.51 (m, 6H), 3.50-3.47 (m, 4H), 3.45-3.43 (m, 4H), 3.25 (br d, J=3.4 Hz, 1H), 3.16 (br s, 1H), 3.06-2.92 (m, 4H), 2.74-2.68 (m, 1H), 2.31 ( s, 3H), 2.14 (dd, J=6.1, 9.4 Hz, 1H), 1.91-1.74 (m, 2H), 1.63 (br dd, J=9.9 , 14.1 Hz, 1H), 1.31-1.10 (m, 2H), 0.96 (s, 9H).

[001420] Synthesis example of (2S,4R)-1-(4-(2-(2-(3-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-( 3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)-2-(3-methylisoxazol-5-yl)butanoyl)-4-hydroxy-N-((S) -1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (representative compound 462)

[001421] Step 1: Preparation of 1,2-bis(2-bromoethoxy)ethane

[001422] To a solution of 2-[2-[2-(2-hydroxyethoxy)ethoxy]ethoxy]ethanol (20 g, 102.97 mmol, 17.70 mL, 1 eq.) and carbon tetrabromide (75.13 g, 226.54 mmol, 2.2 eq.) in tetrahydrofuran (200 ml), triphenylphosphine (58.07 g, 221.39 mmol, 2.15 eq.) was added dropwise over 30 minutes at 0°C. After addition, the mixture was stirred at 20°C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent to obtain a residue. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 50/1 to 20:1). The compound 1-(2-bromoethoxy)-2-[2-(2-bromoethoxy)ethoxy]ethane (30 g, 89.06 mmol, 86% yield, 95% purity) was obtained as a colorless oil.

[001423] Step 2: Preparation of 4-(2-(2-bromoethoxy)ethoxy)-2-(3-methylisoxazol-5-yl)butanoic acid

[001424] To a mixture of 2-(3-methylisoxazol-5-yl)acetic acid (4.09 g, 28.99 mmol, 1 eq.) in tetrahydrofuran (60 ml) was added p-butyllithium (2.5 M, 28 .99 ml, 2.5 eq.) drop by drop at minus 78°C. After addition, the mixture was stirred at 0°C for 0.5 hour. Then 1,2-bis(2-bromoethoxy)ethane (16 g, 57.98 mmol, 2 eq.) was added dropwise at −78°C. The mixture was stirred at minus 78°C for 1 hour. The mixture was then allowed to warm to room temperature (25°C) and stirred at 25°C for 10.5 hours. The mixture was poured into ice water (100 ml), and then a saturated sodium bicarbonate solution was added to make the solution alkaline (pH>8). The solution was extracted with ethyl acetate (100 ml) and the organic layer was removed. A saturated citric acid solution was added to acidify the aqueous phase (pH=4) and the solution was extracted with ethyl acetate (50 ml x 3). The combined organic layers were dried, filtered and concentrated to leave a residue. The residue was purified by semipreparative reverse phase HPLC. The compound 4-[2-(2-bromoethoxy)ethoxy]-2-(3-methylisoxazol-5-yl)butanoic acid (150 mg, 240.94 µmol, 0.8% yield, 54% purity) was obtained as brown gum LC/MS (ESI) m/z: 336.0 [M+1] ⁺ .

[001425] Step 3: Preparation of (2S,4R)-1-(4-(2-(2-bromoethoxy)ethoxy)-2-(3-methylisoxazol-5-yl)butanoyl)-4-hydroxy-N-( (S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001426] To a solution of 4-[2-(2-bromoethoxy)ethoxy]-2-(3-methylisoxazol-5-yl)butanoic acid (150 mg, 446.19 µmol, 1 eq.) and HATU (508.97 mg, 1.34 mmol, 3 eq.) in N,N-dimethylformamide (4 ml) was added (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5 -yl)phenyl]ethyl]pyrrolidine-2-carboxamide (147.88 mg, 446.19 µmol, 1 eq.) and N,N-diisopropylethylamine (230.67 mg, 1.78 mmol, 310.87 µl, 4 eq.). The reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was quenched with water (10 ml) at 0°C, then extracted with ethyl acetate (10 ml × 3). The combined organic layers were washed with brine (10 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a residue. The residue was purified by silica gel column chromatography (dichloromethane/methyl alcohol 10/1). Compound (2S,4R)-1-[4-[2-(2-bromoethoxy)ethoxy]-2-(3-methylisoxazol-5-yl)butanoyl]-4-hydroxy-N-[(1S)-1- [4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (0.13 g, 200.12 µmol, 44% yield) was obtained as a brown gum. LC/MS (ESI) m/z: 651.1 [M+1] ⁺ .

[001427] Step 4: Preparation of (2S,4R)-4-hydroxy-1-(4-(2-(2-(methylamino)ethoxy)ethoxy)-2-(3-methylisoxazol-5-yl)butanoyl)- N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001428] Mixture of (2S,4R)-1-[4-[2-(2-bromoethoxy)ethoxy]-2-(3-methylisoxazol-5-yl)butanoyl]-4-hydroxy-N-[(1S) -1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (0.1 g, 153.94 µmol, 1 eq.) and methylamine/ethyl alcohol (100.00 mmol, 10 ml, purity 30%) was stirred at 60°C for 12 hours. The mixture was concentrated to give the product (2S,4R)-4-hydroxy-1-[4-[2-[2-(methylamino)ethoxy]ethoxy]-2-(3-methylisoxazol-5-yl)butanoyl]-N- [(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (90 mg, 150.07 µmol, 97% yield) as a yellow solid. LC/MS (ESI) m/z: 600.2 [M+1] ⁺ .

[001429] Step 5: Preparation of (2S,4R)-1-(4-(2-(2-(3-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7 -(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)-N-methylpropanamido)ethoxy)ethoxy)-2-(3-methylisoxazol-5-yl)butanoyl)-4-hydroxy-N-(( S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001430] To a solution of (2S,4R)-4-hydroxy-1-[4-[2-[2-(methylamino)ethoxy]ethoxy]-2-(3-methylisoxazol-5-yl)butanoyl]-N- [(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (90 mg, 150.07 µmol, 1 eq.) and 3-[[4-(4 -acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]propanoic acid (88.80 mg, 165.07 µmol, 1. 1 equiv.) in N,N-dimethylformamide (3 ml), 1-hydroxybenzotriazole (40.55 mg, 300.13 µmol, 2 equiv.) and N,N-diisopropylethylamine (77.58 mg, 600.26 µmol, 104.55 µl, 4 eq.). The mixture was stirred at 25°C for 0.15 hours. 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (57.54 mg, 300.13 μmol, 2 eq.) was added to the mixture. The resulting mixture was stirred at 25°C for 2 hours. The reaction mixture was quenched with water (15 ml) at 0°C, then extracted with ethyl acetate (10 ml × 3). The combined organic layers were washed with brine (15 ml), dried over anhydrous sodium sulfate, filtered and concentrated to leave a residue. The residue was purified by semipreparative reverse phase HPLC. Compound (2S,4R)-1-[4-[2-[2-[3-[[4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy- 1-naphthyl)quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]-2-(3-methylisoxazol-5-yl)butanoyl]-4-hydroxy-N-[(1S)-1- [4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (74.7 mg, 66.05 µmol, 44% yield, 99% purity) was obtained as a yellow solid. LC/MS (ESI) m/z: 1119.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 8.90-8.75 (m, 1H), 7.82-7.65 (m, 2H), 7.45-7.29 (m, 5H ), 7.23 (br s, 2H), 7.18-7.09 (m, 1H), 7.03 (br s, 1H), 6.27-6.04 (m, 1H), 5, 03-4.93 (m, 1H), 4.61-4.52 (m, 1H), 4.48-4.16 (m, 2H), 3.84-3.41 (m, 21H), 3.31 (br d, J=1.5 Hz, 2H), 3.07 (br d, J=9.5 Hz, 1H), 2.99-2.65 (m, 4H), 2.43 (s, 3H), 2.25-1.91 (m, 10H), 1.46 (br d, J=6.5 Hz, 3H), 1.38-1.19 (m, 2H), 0 .88 (br d, J=9.2 Hz, 1H).

[001431] Synthesis example of (2S,4R)-1-(1-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazoline -2-yl)amino)-4-methyl-16-(3-methylisoxazol-5-yl)-3-oxo-7,10,13-trioxa-4-azaheptadecan-17-oil)-4-hydroxy-N -((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (representative compound 463)

[001432] Step 1: Preparation of 1-bromo-2-(2-(2-(2-bromoethoxy)ethoxy)ethoxy)ethane

[001433] Triphenylphosphine (112.66 g, 429.51 mmol, 2.15 eq.) and carbon tetrabromide (145.75 g, 439.49 mmol, 2.2 eq.). The mixture was stirred at 20°C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 100/1 to 40/1). The compound 1,2-bis(2-bromoethoxy)ethane (50 g, 173.93 mmol, 87% yield) was obtained as a yellow oil.

[001434] Step 2: Preparation of 4-(2-(2-(2-bromoethoxy)ethoxy)ethoxy)-2-(3-methylisoxazol-5-yl)butanoic acid

[001435] To a mixture of 2-(3-methylisoxazol-5-yl)acetic acid (1.8 g, 12.75 mmol, 1 eq.) in tetrahydrofuran (20 ml) was added p-butyllithium (2.5 M, 12 .75 ml, 2.5 equiv.) drop by drop at minus 78°C. After addition, the mixture was stirred at 0°C for 0.5 hour. Then, 1-(2-bromoethoxy)-2-[2-(2-bromoethoxy)ethoxy]ethane (8.16 g, 25.51 mmol, 2 eq.) was added dropwise at minus 78°C. The mixture was stirred at minus 78°C for 1 hour. The mixture was then allowed to warm to room temperature (25°C) and stirred at 25°C for 10.5 hours. The mixture was poured into ice water (50 ml), and then a saturated sodium bicarbonate solution was added to make the solution alkaline (pH>8). The solution was extracted with ethyl acetate (30 ml) and the organic layer was removed. A saturated citric acid solution was added to acidify the aqueous phase (pH=4) and the solution was extracted with ethyl acetate (30 ml x 3). The combined organic layers were dried, filtered and concentrated to leave a residue. The residue was purified by HPLC. The compound 4-[2-[2-(2-bromoethoxy)ethoxy]ethoxy]-2-(3-methylisoxazol-5-yl)butanoic acid (0.15 g, 394.50 μmol, 3% yield) was obtained as brown gum. LC/MS (ESI) m/z: 380.1 [M+1] ⁺ .

[001436] Step 3: Preparation of (2S,4R)-1-(4-(2-(2-(2-bromoethoxy)ethoxy)ethoxy)-2-(3-methylisoxazol-5-yl)butanoyl)-4- hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001437] To a solution of 4-[2-[2-(2-bromoethoxy)ethoxy]ethoxy]-2-(3-methylisoxazol-5-yl)butanoic acid (0.12 g, 315.60 µmol, 1 eq. ) and HATU (360.00 mg, 946.79 µmol, 3 eq.) in N,N-dimethylformamide (4 ml) was added (2S,4R)-4-hydroxy-N-[(1S)-1-[4 -(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (104.60 mg, 315.60 µmol, 1 eq.) and N,N-diisopropylethylamine (163.16 mg, 1.26 mmol , 219.89 µl, 4 eq.). The reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was quenched with water (10 ml) at 0°C, then extracted with ethyl acetate (10 ml × 3). The combined organic layers were washed with brine (10 ml), dried over anhydrous sodium sulfate, filtered and concentrated to leave a residue. The residue was purified by silica gel column chromatography (dichloromethane/methyl alcohol 10/1). Compound (2S,4R)-1-[4-[2-[2-(2-bromoethoxy)ethoxy]ethoxy]-2-(3-methylisoxazol-5-yl)butanoyl]-4-hydroxy-N-[( 1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (0.13 g, 187.42 μmol, 59% yield) was obtained as a brown gum. LC/MS (ESI) m/z: 695.2 [M+1] ⁺ .

[001438] Step 4: Preparation of (2S,4R)-4-hydroxy-1-(14-(3-methylisoxazol-5-yl)-5,8,11-trioxa-2-azapentadecan-15-oil)-N -((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001439] Mixture of (2S,4R)-1-[4-[2-[2-(2-bromoethoxy)ethoxy]ethoxy]-2-(3-methylisoxazol-5-yl)butanoyl]-4-hydroxy-N -[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (0.1 g, 144.17 µmol, 1 eq.) and methylamine/ethyl alcohol ( 144.17 µmol, 10 ml, 30% purity, 1 eq.) was stirred at 60°C for 12 hours. The reaction was concentrated to give the product, (2S,4R)-4-hydroxy-1-[4-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]-2-(3-methylisoxazol-5-yl )butanoyl]-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (80 mg, 124.26 µmol, 86% yield) as yellow solid matter. LC/MS (ESI) m/z: 644.3 [M+1] ⁺ .

[001440] Step 5: Preparation of (2S,4R)-1-(1-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl) )quinazolin-2-yl)amino)-4-methyl-16-(3-methylisoxazol-5-yl)-3-oxo-7,10,13-trioxa-4-azaheptadecan-17-oil)-4-hydroxy -N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001441] To a solution of (2S,4R)-4-hydroxy-1-[4-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]-2-(3-methylisoxazol-5-yl) butanoyl]-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (80 mg, 124.26 µmol, 1 eq.) and 3-[ [4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]propanoic acid (73.54 mg, 136, 69 µmol, 1.1 eq.) in N,N-dimethylformamide (3 ml), 1-hydroxybenzotriazole (33.58 mg, 248.53 µmol, 2 eq.) and N,N-diisopropylethylamine (64.24 mg, 497.05 µmol, 86.58 µl, 4 eq.). The mixture was stirred at 25°C for 0.15 hours. 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (47.64 mg, 248.53 μmol, 2 eq.) was added to the mixture. The resulting mixture was stirred at 25°C for 2 hours. The reaction mixture was quenched with water (15 ml) at 0°C, then extracted with ethyl acetate (10 ml × 3). The combined organic layers were washed with brine (15 ml), dried over anhydrous sodium sulfate, filtered and concentrated to leave a residue. The residue was purified by semipreparative reverse phase HPLC. Compound (2S,4R)-1-[4-[2-[2-[2-[3-[[4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3 -hydroxy-1-naphthyl)quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethoxy]-2-(3-methylisoxazol-5-yl)butanoyl]-4-hydroxy-N-[( 1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (62.4 mg, 53.08 µmol, 42% yield, 99% purity) was obtained as a yellow solid substances. LC/MS (ESI) m/z: 1163.3 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 8.90-8.83 (m, 1H), 7.78-7.70 (m, 2H), 7.41-7.35 (m, 5H ), 7.23 (br s, 2H), 7.20-7.11 (m, 1H), 7.03 (br s, 1H), 6.23-6.13 (m, 1H), 5, 00 (td, J=7.0, 18.7 Hz, 1H), 4.61-4.54 (m, 1H), 4.48-4.27 (m, 2H), 3.75 (br d , J=16.5 Hz, 8H), 3.58-3.51 (m, 8H), 3.10 (br d, J=7.6 Hz, 1H), 2.91 (br d, J= 7.0 Hz, 2H), 2.83-2.70 (m, 2H), 2.65 (s, 1H), 2.45 (d, J=2.4 Hz, 3H), 2.24- 2.10 (m, 10H), 1.47 (br dd, J=3.1, 6.5 Hz, 3H), 1.28 (s, 1H), 0.88 (br d, J=8, 9 Hz, 1H).

[001442] Synthesis example of (2S,4R)-1-((2S)-20-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1 -yl)quinazolin-2-yl)oxy)-2-(tert-butyl)-18-methyl-4-oxo-6,9,12,15-tetraoxa-3,18-diazaicosanoyl)-4-hydroxy-N -((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (representative compound 473)

[001443] Step 1: Preparation of tert-butyl 4-(7-bromo-6-chloro-8-fluoro-2-(2-hydroxyethoxy)quinazolin-4-yl)piperazine-1-carboxylate

[001444] To a solution of ethylene glycol (1.29 g, 20.83 mmol, 1.2 mL, 5 eq.) in N,N-dimethylformamide (10 mL) was added sodium hydride (200 mg, 5.00 mmol, 60% in mineral oil, 1.2 eq.) at 0°C, the mixture was stirred at 0°C for 30 minutes, and then tert-butyl 4-(7-bromo-2,6-dichloro-8-fluoroquinazoline -4-yl)piperazine-1-carboxylate (2 g, 4.17 mmol, 1 eq.) in N,N-dimethylformamide (2 ml) was added at 0°C, the mixture was stirred at 20°C for 1 hour . The mixture was quenched with water (100 ml) and extracted with ethyl acetate (100 ml, twice), the organic phase was dried with anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the crude product. This crude product was purified by flash (0:1 to 1:1 ethyl acetate:petroleum ether) to give the compound tert-butyl 4-[7-bromo-6-chloro-8-fluoro-2-(2-hydroxyethoxy)quinazoline -4-yl]piperazine-1-carboxylate (1.15 g, 2.09 mmol, 50% yield), 92% purity) as a pale yellow solid. LC/MS (ESI) m/z: 506.9 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.72 (d, J=2.0 Hz, 1H), 4.66-4.54 (m, 2H), 4.04-3.97 (m , 2H), 3.86-3.76 (m, 4H), 3.69-3.60 (m, 4H), 1.50 (s, 9H).

[001445] Step 2: Preparation of tert-butyl 4-(7-bromo-6-chloro-8-fluoro-2-(((S)-19-((2S,4R)-4-hydroxy-2-(( (S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-3,20,20-trimethyl-17-oxo-6,9,12,15 -tetraoxa-3,18-diazagenicosyl)oxy)quinazolin-4-yl)piperazine-1-carboxylate

[001446] To a solution of tert-butyl 4-[7-bromo-6-chloro-8-fluoro-2-(2-hydroxyethoxy)quinazolin-4-yl]piperazine-1-carboxylate (200 mg, 0.40 mmol, 1 eq.) in dichloromethane (3 ml) and dimethyl sulfoxide (1.00 g, 12.80 mmol, 1 ml, 32.37 eq.), triethylamine (200 mg, 1.98 mmol, 5 eq.) was added, and then pyridine sulfur trioxide (189 mg, 1.19 mmol, 3 eq.), the mixture was stirred at 20°C for 10 hours. The mixture was loaded with (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]ethoxy] acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (371 mg, 0.54 mmol, 0.9 equiv.), triethylamine (121 mg, 1.19 mmol, 2 equiv.), dichloromethane (4 ml) and dimethyl sulfoxide (5 ml) at 20°C under nitrogen atmosphere. The mixture was stirred at 20°C for 30 minutes. Sodium triacetoxyborohydride (631 mg, 2.98 mmol, 5 eq.) was then added. The mixture was stirred at 20°C for 1 hour. The mixture was concentrated in vacuo. The reaction mixture was purified by semipreparative reverse phase HPLC to give the compound tert-butyl 4-[7-bromo-6-chloro-8-fluoro-2-[2-[2-[2-[2-[2-[2-[ [(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidin-1- carbonyl]-2,2-dimethyl-propyl]amino]-2-oxoethoxy]ethoxy]ethoxy]ethoxy]ethyl-methyl-amino]ethoxy]quinazolin-4-yl]piperazin-1-carboxylate (250 mg, 0.20 mmol, yield 34%, formate) as a white solid. LC/MS (ESI) m/z: 590.5 [M/2+1] ⁺ .

[001447] Step 3: Preparation of tert-butyl 4-(6-chloro-8-fluoro-2-(((S)-19-((2S,4R)-4-hydroxy-2-(((S)- 1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-3,20,20-trimethyl-17-oxo-6,9,12,15-tetraoxa-3 ,18-diazagenicosyl)oxy)-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazine-1-carboxylate

[001448] Mixture of tert-butyl 4-[7-bromo-6-chloro-8-fluoro-2-[2-[2-[2-[2-[2-[2-[[(1S)-1- [(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2- dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy]ethoxy]ethoxy]ethyl-methyl-amino]ethoxy]quinazolin-4-yl]piperazine-1-carboxylate (50 mg, 0.04 mmol, 1 eq. ), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene-2-ol (17 mg, 0.06 mmol, 1.5 eq.), [2 -(2-aminophenyl)phenyl]palladium(1+); dicyclohexyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane; methanesulfonate (4 mg, 0.004 mmol, 0.1 eq.) and potassium phosphate (1.5 M in H ₂ O, 0.08 ml, 3.0 eq.) in tetrahydrofuran (3 ml) were degassed and purged with nitrogen (3 times), and then the mixture was stirred at a temperature of 65°C for 12 hours under a nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative thin layer chromatography (silica gel plate, dichloromethane/methanol 7/1) to give tert-butyl 4-[6-chloro-8-fluoro-2-[2-[2-[2-[2-[ 2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]ethoxy]ethoxy]ethoxy]ethyl-methyl-amino]ethoxy]-7-(3-hydroxy-1- naphthyl)quinazolin-4-yl]piperazin-1-carboxylate (40 mg, 0.03 mmol, 76% yield) as a yellow solid. LC/MS (ESI) m/z: 1244.0 [M+1] ⁺ .

[001449] Step 4: Preparation of (2S,4R)-1-((2S)-2-(tert-butyl)-20-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl )-4-(piperazin-1-yl)quinazolin-2-yl)oxy)-18-methyl-4-oxo-6,9,12,15-tetraoxa-3,18-diazaicosanoyl)-4-hydroxy-N -((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001450] To a solution of tert-butyl 4-[6-chloro-8-fluoro-2-[2-[2-[2-[2-[2-[2-[[(1S)-1-[(2S ,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl ]amino]-2-oxo-ethoxy]ethoxy]ethoxy]ethoxy]ethyl-methyl-amino]ethoxy]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (40 mg , 0.03 mmol, 1 eq.) in dichloromethane (5 ml) was added hydrochloric acid HCl in dioxane (4 M, 3 ml). The mixture was stirred at 20°C for 0.5 hour. The reaction mixture was concentrated under reduced pressure to give the compound (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[2-[6-chloro-8- fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxyethyl-methyl-amino]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3 -dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (40 mg, hydrochloride) as a yellow solid substances. LC/MS (EST) m/z: 1143.6 [M+1] ⁺ .

[001451] Step 5: Preparation of (2S,4R)-1-((2S)-20-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene -1-yl)quinazolin-2-yl)oxy)-2-(tert-butyl)-18-methyl-4-oxo-6,9,12,15-tetraoxa-3,18-diazaicosanoyl)-4-hydroxy -N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001452] Mixture of (2S,4R)-1-[(2S)2-[[2-[2-[2-[2-[2-[2-[6-chloro-8-fluoro-7-(3 -hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxyethyl-methyl-amino]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]- 4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (40 mg, 0.033 mmol, 1 eq., hydrochloride) and 2, 6-lutidine (106 mg, 0.99 mmol, 30 eq.) in dichloromethane (5 ml) was cooled to minus 78°C, then prop-2-enoyl chloride (3 mg, 0.033 mmol, 1 eq.) in dichloromethane was added ( 1 ml). The mixture was stirred at a temperature of minus 78°C for 0.5 hours. The reaction mixture was quenched with saturated sodium bicarbonate solution (20 ml), then extracted with dichloromethane (20 ml x 2). The combined organic layers were washed with brine (20 ml×1), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by semipreparative reverse phase HPLC to give (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[2-[6-chloro-8 -fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]oxyethyl-methyl-amino]ethoxy]ethoxy]ethoxy]ethoxy] acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (25 .6 mg, 0.019 mmol, yield 56%, purity 95%, trifluoroacetic acid salt) as a yellow solid. LC/MS (ESI) m/z: 589.9 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 9.01-8.89 (m, 1H), 8.17-8.05 (m, 1H), 7.76 (d, J=8.4 Hz, 1H), 7.58-7.38 (m, 5H), 7.28 (d, J=2.4 Hz, 1H), 7.25-7.16 (m, 2H), 7.05 (d, J=2.4 Hz, 1H), 6.89-6.73 (m, 1H), 6.36-6.23 (m, 1H), 5.86-5.75 (m, 1H ), 5.04-4.94 (m, 1H), 4.82-4.75 (m, 1H), 4.69-4.63 (m, 1H), 4.58-4.51 (m , 1H), 4.46-4.31 (m, 1H), 4.16-4.06 (m, 4H), 4.05-3.99 (m, 2H), 3.98-3.89 (m, 4H), 3.88-3.80 (m, 3H), 3.80-3.39 (m, 18H), 3.11-2.99 (m, 3H), 2.51-2 .42 (m, 3H), 2.40-2.13 (m, 1H), 2.03-1.90 (m, 1H), 1.60-1.43 (m, 3H), 1.07 -0.96 (m, 9H).

[001453] Synthesis example of (2S,4R)-1-((2S)-2-(2-(4-(2-(2-(4-((4-(4-acryloylpiperazin-1-yl)-6 -chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)piperidin-1-yl)ethoxy)ethyl)piperazin-1-yl)acetamido)-3,3-dimethylbutanoyl )-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (typical compound 475)

[001454] Step 1: Preparation of 2-(2,2-diethoxyethoxy)ethan-1-ol

[001455] A 100 mL three-neck flask equipped with a thermometer, stirrer, and reflux condenser was charged with ethylene glycol (9.45 g, 152.23 mmol, 8.5 mL, 2 eq.). The flask was cooled in an ice bath. Potassium hydroxide (6.41 g, 114.17 mmol, 1.5 eq.) was added slowly at 0-50°C. 2-Bromo-1,1-diethoxyethane (15 g, 76.12 mmol, 11.45 mL, 1 eq.) was then added and the reaction mixture was heated to 110° C. for 20 hours. The reaction mixture was cooled to 20°C, then water (30 ml) was added. The pH of the mixture was adjusted to 8 by adding 1 N hydrochloric acid. The mixture was then extracted with ethyl acetate (50 ml x 5). The organic layer was dried over sodium sulfate and then concentrated in vacuo to obtain a residue. The residue was purified by silica gel column chromatography (5-80% ethyl acetate in petroleum ether) to give 2-(2,2-diethoxyethoxy)ethanol (3.71 g, 20.82 mmol, 27% yield) as a light yellow oil . ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.65 (t, J=5.2 Hz, 1H), 3.78-3.68 (m, 4H), 3.67-3.62 (m , 2H), 3.62-3.54 (m, 4H), 2.43 (br s, 1H), 1.24 (t, J=7.1 Hz, 6H).

[001456] Step 2: Preparation of 2-(2,2-diethoxyethoxy)ethyl 4-methylbenzenesulfonate

[001457] To a solution of 2-(2,2-diethoxyethoxy)ethanol (300 mg, 1.68 mmol, 1 eq.) in dichloromethane (6 mL) was added triethylamine (511 mg, 5.05 mmol, 0.7 mL, 3 eq.) and 4-methylbenzene-1-sulfonyl chloride (481 mg, 2.52 mmol, 1.5 eq.). The reaction mixture was stirred at 25°C for 20 hours. The reaction mixture was concentrated in vacuo to obtain a residue. The residue was purified by silica gel column chromatography (5-15% ethyl acetate in petroleum ether) to give 2-(2,2-diethoxyethoxy)ethyl 4-methylbenzenesulfonate (404 mg, 1.22 mmol, 72% yield) as a colorless oil.

[001458] Step 3: Preparation of ethyl 2-(4-(2-(2,2-diethoxyethoxy)ethyl)piperazin-1-yl)acetate

[001459] To a solution of ethyl 2-piperazin-1-ylacetate (1.06 g, 5.08 mmol, 1.3 eq., hydrochloric acid) and 2-(2,2-diethoxyethoxy)ethyl 4-methylbenzenesulfonate (1, 3 g, 3.91 mmol, 1 eq.) in acetonitrile (10 ml) was added potassium carbonate (1.62 g, 11.76 mmol, 3.01 eq.). The mixture was stirred at 80°C for 12 hours. Water (10 ml) was added, then the mixture was extracted with ethyl acetate (20 ml x 3), the mixture was dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative HPLC. The compound ethyl 2-[4-[2-(2,2-diethoxyethoxy)ethyl]piperazin-1-yl]acetate (500 mg, 1.50 mmol, 38% yield) was obtained as a yellow oil. LC/MS (ESI) m/z: 333.6 [M+1] ⁺ .

[001460] Step 4: Preparation of 2-(4-(2-(2,2-diethoxyethoxy)ethyl)piperazin-1-yl)acetic acid

[001461] To a solution of ethyl 2-[4-[2-(2,2-diethoxyethoxy)ethyl]piperazin-1-yl]acetate (400 mg, 1.20 mmol, 1 eq.) in a mixture of tetrahydrofuran (4 ml) and methanol (2 ml), water (2.5 ml) and lithium hydroxide monohydrate (101 mg, 2.41 mmol, 2 eq.) were added. The reaction mixture was stirred at 20°C for 2 hours. 2 M sulfuric acid was added to adjust the pH of the mixture to 7. The mixture was then concentrated in vacuo to remove most of the organic solvents and then lyophilized. Methanol (2 ml) and dichloromethane (20 ml) were added. The mixture was filtered and the filtrate was concentrated in vacuo to give 2-[4-[2-(2,2-diethoxyethoxy)ethyl]piperazin-1-yl]acetic acid (320 mg, 1.05 mmol, 87% yield) as light - yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 4.62 (t, J=5.1 Hz, 1H), 3.74-3.62 (m, 4H), 3.56 (qd, J=7 ,1, 9.4 Hz, 2H), 3.49 (br s, 3H), 3.33 (br s, 2H), 3.05 (br s, 4H), 2.78 (br s, 4H) , 2.67 (br s, 2H), 1.22 (t, J=1.0 Hz, 6H).

[001462] Step 5: Preparation of (2S,4R)-1-((S)-2-(2-(4-(2-(2,2-diethoxyethoxy)ethyl)piperazin-1-yl)acetamido)-3 ,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001463] To a solution of 2-[4-[2-(2,2-diethoxyethoxy)ethyl]piperazin-1-yl]acetic acid (152 mg, 0.50 mmol, 1.2 eq.), (2S,4R )-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl ]pyrrolidine-2-carboxamide (200 mg, 0.42 mmol, 1 eq., hydrochloride), hydroxybenzotriazole (73 mg, 0.54 mmol, 1.3 eq.) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (104 mg, 0.54 mmol, 1.3 eq.) in N,N-dimethylformamide (4 ml) was added N,N-diisopropylethylamine (161 mg, 1.25 mmol, 0.22 ml, 3 eq.) . The reaction mixture was stirred at 20°C for 4 hours. The reaction mixture was then stirred at 20°C for another 14 hours. Water (20 ml) was added and the mixture was extracted with ethyl acetate (20 ml x 2). The organic layer was dried over sodium sulfate and then concentrated in vacuo to obtain a residue. The residue was purified by preparative TLC (10% methanol in dichloromethane) to give (2S,4R)-1-[(2S)-2-[[2-[4-[2-(2,2-diethoxyethoxy)ethyl]piperazine-1 -yl]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidin-2- carboxamide (192 mg, 0.24 mmol, yield 58%, purity 92%) as a colorless gum. LC/MS (ESI) m/z: 731.5 [M+1] ⁺ .

[001464] Step 6: Preparation of (2S,4R)-1-((S)-3,3-dimethyl-2-(2-(4-(2-(2-oxoethoxy)ethyl)piperazin-1-yl) acetamido)butanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001465] To a solution of (2S,4R)-1-[(2S)-2-[[2-[4-[2-(2,2-diethoxyethoxy)ethyl]piperazin-1-yl]acetyl]amino]- 3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (100 mg, 0.14 mmol, 1 eq.) in tetrahydrofuran (3 ml), sulfuric acid (2 M, 0.5 ml, 7.31 eq.) was added. The reaction mixture was stirred at 60°C for 1 hour. Solid sodium sulfate was added to the mixture to adjust the pH to 7. Tetrahydrofuran (30 ml) and methanol (2 ml) were then added and the mixture was dried over sodium sulfate. The solution was concentrated in vacuo to give (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[4-[2-(2-oxoethoxy)ethyl]piperazin-1-yl] acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (73 mg) as a colorless gum. LC/MS (ESI) m/z: 679.4 [M+23] ⁺ .

[001466] Step 7: Preparation of 2-(trimethylsilyl)ethyl 4-hydroxypiperidine-1-carboxylate

[001467] To a solution of piperidin-4-ol (1.17 g, 11.57 mmol, 3 eq.) in tetrahydrofuran (10 mL) was added triethylamine (1.17 g, 11.57 mmol, 1.61 mL, 3 eq.), (2,5-dioxopyrrolidin-1-yl)-2-trimethylsilylethyl carbonate (1 g, 3.86 mmol, 1 eq.) was added to the mixture, the reaction was stirred at 25°C for 2 h. the mixture was quenched with water (30 ml) and extracted with ethyl acetate (20 ml x 2). The combined organic layers were washed with brine (30 ml), dried over sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 5:1 to 0:1) to give 2-trimethylsilylethyl 4-hydroxypiperidine-1-carboxylate (985 mg) as a colorless oil. ¹ H-NMR (400 MHz, CD ₃ OD) δ 4.22-4.16 (m, 4H), 3.71-3.67 (m, 10H), 3.60-3.56 (m, 1H ), 3.31-3.13 (m, 1H), 1.81-1.61 (m, 2H), 1.50-1.47 (m, 2H), 1.47 (s, 9H), 1.27 (t, J=7.2 Hz, 3H).

[001468] Step 8: Preparation of tert-butyl 4-(7-bromo-6-chloro-8-fluoro-2-((1-((2-(trimethylsilyl)ethoxy)carbonyl)piperidin-4-yl)oxy) quinazolin-4-yl)piperazine-1-carboxylate

[001469] Tert-Butyl 4-(7-bromo-2,6-dichloro-8-fluoro-quinazolin-4-yl)piperazin-1-carboxylate mixture (500 mg, 1.04 mmol, 1 eq.), 2 -trimethylsilylethyl 4-hydroxypiperidine-1-carboxylate (511 mg, 2.08 mmol, 2 eq.), potassium (432 mg, 3.12 mmol, 3 eq.) and 1,4-diazabicyclo[2.2,2]octane ( 12 mg, 0.10 mmol, 0.1 eq.) in acetonitrile (25 ml) was heated at 85°C for 14 hours. The reaction mixture was filtered and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by silica gel column chromatography (0-15% ethyl acetate in petroleum ether) to give tert-butyl 4-[7-bromo-6-chloro-8-fluoro-2-[[1-(2-trimethylsilylethoxycarbonyl)-4 -piperidyl]oxy]quinazolin-4-yl]piperazine-1-carboxylate (240 mg, 0.35 mmol, 33% yield) as a yellow solid.

[001470] Step 9: Preparation of tert-butyl 4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-2-((1-((2-(trimethylsilyl)ethoxy)carbonyl) piperidin-4-yl)oxy)quinazolin-4-yl)piperazine-1-carboxylate

[001471] To a solution of tert-butyl 4-[7-bromo-6-chloro-8-fluoro-2-[[1-(2-trimethylsilylethoxycarbonyl)-4-piperidyl]oxy]quinazolin-4-yl]piperazin-1 -carboxylate (230 mg, 333.77 µmol, 1 eq.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene-2-ol (117 mg, 0.43 mmol, 1.3 eq.) in tetrahydrofuran (8 ml), potassium phosphate (1.5 M, 0.67 ml, 3 eq.) and methanesulfonato(2-dicyclohexylphosphino-2,4,6-tri- i-propyl-1,1-biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(ii) (28 mg, 0.33 mol, 0.1 eq.). The reaction mixture was degassed and charged with nitrogen 3 times and then heated to 65°C for 16 hours. Ethyl acetate (30 ml) was added and the mixture was washed with water (30 ml). The organic layer was dried over sodium sulfate and then concentrated in vacuo to obtain a residue. The residue was purified by preparative TLC (60% ethyl acetate in petroleum ether) to give tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[[1-(2-trimethylsilylethoxycarbonyl )-4-piperidyl]oxy]quinazolin-4-yl]piperazine-1-carboxylate (200 mg) as a light yellow solid. LC/MS (ESI) m/z: 752.1 [M+1] ⁺ .

[001472] Step 10: Preparation of tert-butyl 4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-2-(piperidin-4-yloxy)quinazolin-4-yl)piperazin- 1-carboxylate

[001473] To a solution of tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[[1-(2-trimethylsilylethoxycarbonyl)-4-piperidyl]oxy]quinazoline -4-yl]piperazine-1-carboxylate (180 mg, 0.24 mmol, 1 eq.) in tetrahydrofuran (4 ml) was added tetrabutylammonium fluoride (1 M, 0.26 ml, 1.1 eq.). The reaction mixture was heated to 50°C for 14 hours. The reaction solution was concentrated in vacuo to obtain a residue. The residue was purified by preparative HPLC to give tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-(4-piperidyloxy)quinazolin-4-yl]piperazine-1-carboxylate (130 mg, 0.18 mmol, 75% yield, trifluoroacetate) as a light yellow solid. LC/MS (EST) m/z: 608.3 [M+1] ⁺ .

[001474] Step 11: Preparation of tert-butyl 4-(6-chloro-8-fluoro-2-((1-(2-(2-(4-(2-(((S)-1-((2S ,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1 -oxobutan-2-yl)amino)-2-oxoethyl)piperazin-1-yl)ethoxy)ethyl)piperidin-4-yl)oxy)-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl) piperazine-1-carboxylate

[001475] To a solution of tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-(4-piperidyloxy)quinazolin-4-yl]piperazine-1-carboxylate ( 70 mg, 0.1 mmol, 1 eq., trifluoroacetate) in methanol (1 ml) was added sodium acetate (24 mg, 0.29 mmol, 3 eq.) at 20°C. The reaction mixture was stirred at 20°C for 20 minutes. Acetic acid (12 mg, 0.2 mmol, 2 eq.) and a solution of (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[4-[2- (2-oxoethoxy)ethyl]piperazin-1-yl]acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine -2-carboxamide (73 mg, 0.11 mmol, 1.15 eq.) in dichloromethane (3 ml) and the mixture was cooled to 0°C. Sodium cyanoborohydride (9 mg, 0.15 mmol, 1.5 eq.) was added and the mixture was stirred at 20°C for 14 hours. The solution was purified by preparative TLC (8% methanol in dichloromethane) to give tert-butyl 4-[6-chloro-8-fluoro-2-[[1-[2-[2-[4-[2-[[(1S) -1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2 ,2-dimethyl-propyl]amino]-2-oxo-ethyl]piperazin-1-yl]ethoxy]ethyl]-4-piperidyl]oxy]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl ]piperazine-1-carboxylate (53 mg, 0.04 mmol, 38% yield, 87% purity) as a colorless oil. LC/MS (ESI) m/z: 1238.6 [M+1] ⁺ .

[001476] Step 12: Preparation of (2S,4R)-1-((2S)-2-(2-(4-(2-(2-(4-((6-chloro-8-fluoro-7-( 3-hydroxynaphthalene-1-yl)-4-(piperazin-1-yl)quinazolin-2-yl)oxy)piperidin-1-yl)ethoxy)ethyl)piperazin-1-yl)acetamido)-3,3-dimethylbutanoyl )-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001477] To a solution of tert-butyl 4-[6-chloro-8-fluoro-2-[[1-[2-[2-[4-[2-[[(1S)-1-[(2S,4R )-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino ]-2-oxo-ethyl]piperazin-1-yl]ethoxy]ethyl]-4-piperidyl]oxy]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazin-1-carboxylate (52 mg, 0.04 mmol, 1 eq.) in dichloromethane (3 ml) was added trifluoroacetic acid (0.5 M, 0.08 ml, 1 eq.). The mixture was stirred at 15°C for 1 hour. The reaction mixture was concentrated in vacuo to give (2S,4R)-1-[(2S)-2-[[2-[4-[2-[2-[4-[6-chloro-8-fluoro-7-( 3-hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxy-1-piperidyl]ethoxy]ethyl]piperazin-1-yl]acetyl]amino]-3,3-dimethyl- butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (52 mg, trifluoroacetate) as a colorless oil.

[001478] Step 13: Preparation of (2S,4R)-1-((2S)-2-(2-(4-(2-(2-(4-((4-(4-acryloylpiperazin-1-yl) -6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)oxy)piperidin-1-yl)ethoxy)ethyl)piperazin-1-yl)acetamido)-3,3 -dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001479] To a solution of (2S,4R)-1-[(2S)-2-[[2-[4-[2-[2-[4-[6-chloro-8-fluoro-7-(3- hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]oxy-1-piperidyl]ethoxy]ethyl]piperazin-1-yl]acetyl]amino]-3,3-dimethyl-butanoyl] -4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (52 mg, 41.18 µmol, 1 eq., trifluoroacetate) 2,6-lutidine (176 mg, 1.65 mmol, 40 eq.) was added to N,N-dimethylformamide (1 ml) and dichloromethane (5 ml). The reaction mixture was cooled to minus 70°C. A solution of prop-2-enoyl chloride (2.61 mg, 0.03 mmol, 0.7 eq.) in dichloromethane (0.23 ml) was then added and the mixture was stirred at minus 70° C. for 10 minutes. Dichloromethane (20 ml) and water (15 ml) were added and the mixture was separated. The organic layer was dried over sodium sulfate, then concentrated to obtain a residue. The residue was purified by preparative HPLC to give (2S,4R)-1-[(2S)-2-[[2-[4-[2-[2-[4-[6-chloro-8-fluoro-7-(3 -hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]oxy-1-piperidyl]ethoxy]ethyl]piperazin-1-yl]acetyl]amino]- 3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (10.2 mg, 0.008 mmol, yield 20%, purity 98%) as a white solid. LC/MS (ESI) m/z: 1202.6 [M+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.98 (s, 1H), 8.74-8.38 (m, 1H), 8.27 (s, 2H), 8.00 (s , 1H), 7.80 (d, J=8.3 Hz, 1H), 7.74-7.49 (m, 1H), 7.46-7.39 (m, 3H), 7.38- 7.32 (m, 2H), 7.28 (d, J=2.3 Hz, 1H), 7.25-7.16 (m, 2H), 7.06 (d, J=2.3 Hz , 1H), 6.83 (dd, J=10.5, 16.6 Hz, 1H), 6.18 (dd, J=2.3, 16.7 Hz, 1H), 5.78-5, 70 (m, 1H), 5.03 (td, J=4.4, 8.3 Hz, 1H), 4.88 (quin, J=7.0 Hz, 1H), 4.53-4.33 (m, 2H), 4.30-4.20 (m, 1H), 3.92 (br d, J=2.5 Hz, 4H), 3.85 (br s, 2H), 3.77 ( br s, 2H), 3.60-3.55 (m, 4H), 3.49 (br s, 10H), 3.02 (br d, J=16.1 Hz, 1H), 2.91- 2.83 (m, 2H), 2.78 (br s, 2H), 2.47 (br s, 4H), 2.45 (s, 3H), 2.30-2.22 (m, 2H) , 2.10-1.95 (m, 3H), 1.80-1.62 (m, 1H), 1.80-1.62 (m, 2H), 1.48-1.33 (m, 3H), 0.90 (s, 8H).

[001480] Example of the synthesis of (2S,4R)-1-((2S)-14-(4-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3- hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)phenoxy)-2-(tert-butyl)-4-oxo-6,9,12-trioxa-3-azatetradecanoyl)-4-hydroxy-N-(( S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (representative compound 483)

[001481] Step 1: Preparation of 2-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)ethyl 4-methylbenzenesulfonate

[001482] To a solution of 2-(2-(2-hydroxyethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (32.70 g, 107.44 mmol, 1.00 eq.) in dichloromethane (300 ml) was added 4-methylbenzenesulfonic acid; pyridine (2.97 g, 11.82 mmol, 0.11 eq) and then 3,4-dihydro-2H-pyran (14.28 g, 169.75 mmol, 15.5 mL, 1 .58 eq.) at 15°C. The mixture was stirred at 15°C for 3 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent and then diluted with water (300 ml) and extracted with ethyl acetate (300 ml x 2). The combined organic layers were washed with water (300 ml x 2), dried over sodium sulfate, filtered and concentrated under reduced pressure to give the product. Compound 2-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)ethyl 4-methyl benzenesulfonate (37.40 g, 89.92 mmol, 84% yield, 93% purity ) was obtained as a white liquid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.82 (d, J=8.0 Hz, 2H), 7.36 (d, J=8.4 Hz, 2H), 4.64 (t, J=3.2 Hz, 1H), 4.18 (t, J=4.8 Hz, 2H), 3.73-3.53 (m, 12H), 2.47 (s, 3H), 1, 84-1.73 (m, 2H), 1.61-1.52 (m, 4H).

[001483] Step 2: Preparation of 2-((1-phenyl-2,6,9,12-tetraoxatetradecane-14-yl)oxy)tetrahydro-2H-pyran

[001484] To a solution of 2-(2-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (37.00 g, 95.24 mmol, 1.00 eq .) in xylene (370 ml) was added potassium hydroxide (6.41 g, 114.29 mmol, 1.20 eq.) and 3-(benzyloxy)propan-1-ol (15.83 g, 95.24 mmol, 15.1 ml, 1.00 eq.). The mixture was stirred at 130°C for 3 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with ethyl acetate (800 ml). The combined organic layers were washed with water (300 ml x 2), dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash chromatography on silica gel. The compound 2-((1-phenyl-2,6,9,12-tetraoxatetradecane-14-yl)oxy)tetrahydro-2H-pyran (19.00 g, 41.23 mmol, 43% yield, 83% purity) was obtained as a white solid. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.35-7.28 (m, 5H), 4.64 (t, J=3.6 Hz, 1H), 4.51 (s, 2H), 3.88-3.49 (m, 18H), 2.05-1.26 (m, 8H).

[001485] Step 3: Preparation of ethyl 2-(2-(2-(2-(4-nitrophenoxy)ethoxy)ethoxy)ethoxy)acetate

[001486] To a solution of ethyl 2-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]acetate (3 g, 7.68 mmol, 1 eq.) and 4-nitrophenol (1.28 g, 9.22 mmol, 1.2 eq.) in dimethylformamide (10 ml) was added potassium carbonate (2.12 g, 15.37 mmol, 2 eq.). The reaction mixture was stirred at 50°C for 12 hours. The reaction mixture was quenched by adding water (20 ml) and then diluted with water (30 ml), filtered and extracted with ethyl acetate (40 ml x 3). The combined organic layers were washed with brine (80 ml), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC. The desired compound ethyl 2-[2-[2-[2-(4-nitrophenoxy)ethoxy]ethoxy]ethoxy]acetate (1.3 g, 3.64 mmol, 47% yield) was obtained as a light yellow oil. LC/MS (ESI) m/z: 835.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.27-8.15 (m, 2H), 7.04-6.94 (m, 2H), 4.28-4.17 (m, 4H) , 4.14 (s, 2H), 3.95-3.87 (m, 2H), 3.79-3.66 (m, 8H), 1.28 (t, J=7.2 Hz, 3H ).

[001487] Step 4: Preparation of ethyl 2-(2-(2-(2-(4-aminophenoxy)ethoxy)ethoxy)ethoxy)acetate

[001488] To a solution of ethyl 2-[2-[2-[2-(4-nitrophenoxy)ethoxy]ethoxy]ethoxy]acetate (1.3 g, 3.64 mmol, 1 eq.) in ethyl alcohol (10 ml ) palladium on carbon (500 mg, 10% purity) was added. The reaction mixture was stirred at 40°C for 12 hours with hydrogen [0.1 MPa (15 psi)]. The reaction mixture was filtered and concentrated under reduced pressure to give the product, ethyl 2-[2-[2-[2-(4-aminophenoxy)ethoxy]ethoxy]ethoxy]acetate (1.1 g, 3.36 mmol, yield 92 %) in the form of a light yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 6.82-6.73 (m, 2H), 6.69-6.60 (m, 2H), 4.22 (q, J=7.2 Hz , 2H), 4.15 (s, 2H), 4.09-4.03 (m, 2H), 3.86-3.79 (m, 2H), 3.78-3.66 (m, 9H ), 1.34-1.21 (m, 4H).

[001489] Step 5: Preparation of tert-butyl 4-(7-bromo-6-chloro-8-fluoro-2-((4-((11-oxo-3,6,9,12-tetraoxatetradecyl)oxy)phenyl )amino)quinazolin-4-yl)piperazine-1-carboxylate

[001490] To a solution of ethyl 2-[2-[2-[2-(4-aminophenoxy)ethoxy]ethoxy]ethoxy]acetate (545 mg, 1.67 mmol, 2 eq.) and tert-butyl 4-(7 -bromo-2,6-dichloro-8-fluoroquinazolin-4-yl) piperazine-1-carboxylate (400 mg, 0.83 mmol, 1 eq.) in isopropanol (10 ml) was added N,N-diisopropylethylamine ( 431 mg, 3.33 mmol, 4 eq.). The reaction mixture was stirred at 95°C for 12 hours. The reaction mixture was quenched by adding water (20 ml) and then diluted with water (30 ml), filtered and extracted with ethyl acetate (40 ml x 3). The combined organic layers were washed with brine (80 ml), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by chromatography on silica gel (petroleum ether: ethyl acetate in the ratio from 20:1 to 3:1). Desired compound tert-butyl 4-[7-bromo-6-chloro-2-[4-[2-[2-[2-(2-ethoxy-2-oxo-ethoxy)ethoxy]ethoxy]ethoxy]anilino]- 8-fluoroquinazolin-4-yl]piperazin-1-carboxylate (500 mg, 0.65 mmol, 77% yield) was obtained as a yellow solid. LC/MS (ESI) m/z: 772.1 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.66-7.51 (m, 3H), 7.08 (s, 1H), 6.92-6.88 (m, 2H), 6.80 -6.71 (m, 1H), 6.69-6.59 (m, 1H), 4.27-4.18 (m, 3H), 4.18-4.11 (m, 5H), 4 .08-4.04 (m, 1H), 3.89-3.85 (m, 1H), 3.83-3.79 (m, 1H), 3.78-3.60 (m, 13H) , 1.54-1.48 (m, 9H), 1.28 (t, J=7.2 Hz, 3H).

[001491] Step 6: Preparation of tert-butyl 4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-2-((4-((11-oxo-3,6,9 ,12-tetraoxatetradecyl)oxy)phenyl)amino)quinazolin-4-yl)piperazine-1-carboxylate

[001492] To a solution of tert-butyl 4-[7-bromo-6-chloro-2-[4-[2-[2-[2-(2-ethoxy-2-oxo-ethoxy) ethoxy]ethoxy]ethoxy] anilino]-8-fluoroquinazolin-4-yl]piperazin-1-carboxylate (340 mg, 0.44 mmol, 1 eq.) and 4-(4,4,5,5-tetramethyl-1,3,2 -dioxaborolan-2-yl)naphthalene-2-ol (143 mg, 0.53 mmol, 1.2 eq.) in tetrahydrofuran (10 ml) was added potassium phosphate (1.5 M, 0.9 ml, 3 eq. ) and [2-(2-aminophenyl)phenyl]palladium(1+); dicyclohexyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane; methanesulfonate (37 mg, 0.04 mmol, 0.1 eq.). The reaction mixture was stirred at 60°C for 12 hours. The reaction mixture was quenched by adding water (10 ml) and then diluted with water (20 ml) and extracted with ethyl acetate (30 ml x 3). The combined organic layers were washed with brine (30 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (petroleum ether: ethyl acetate in the ratio 1:1). Desired compound tert-butyl 4-[6-chloro-2-[4-[2-[2-[2-(2-ethoxy-2-oxo-ethoxy)ethoxy]ethoxy]ethoxy]anilino]-8-fluoro- 7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (122 mg, 0.15 mmol, 33% yield) was obtained as a white solid. LC/MS (ESI) m/z: 834.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.76 (d, J=8.0 Hz, 1H), 7.70 (s, 1H), 7.54 (d, J=7.8 Hz, 2H), 7.43 (t, J=7.6 Hz, 1H), 7.38-7.31 (m, 2H), 7.26-7.20 (m, 2H), 7.15 (s , 1H), 6.90 (d, J=8.8 Hz, 2H), 4.17-4.09 (m, 5H), 3.86 (t, J=4.4 Hz, 2H), 3 .80-3.60 (m, 16H), 1.52 (s, 9H), 1.33-1.22 (m, 3H).

[001493] Step 7: Preparation of 2-(2-(2-(2-(4-((4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-6-chloro-8-fluoro-7- (3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)phenoxy)ethoxy)ethoxy)ethoxy)acetic acid

[001494] To a solution of tert-butyl 4-[6-chloro-2-[4-[2-[2-[2-(2-ethoxy-2-oxo-ethoxy)ethoxy]ethoxy]ethoxy]anilino]-8 -fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (180 mg, 0.21 mmol, 1 eq.) in tetrahydrofuran (5 ml) and water (2 ml) lithium hydroxide monohydrate (18 mg, 0.43 mmol, 2 eq.) was added. The reaction mixture was stirred at 15°C for 12 hours. The residue was adjusted to pH=4~5 with 1M hydrogen chloride, then extracted with ethyl acetate (30 ml×2). The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure to give the product, compound 2-[2-[2-[2-[4-[[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6 -chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]phenoxy]ethoxy]ethoxy]ethoxy]acetic acid (170 mg, 0.2 mmol, 92% yield, purity 94%) as a yellow solid. LC/MS (ESI) m/z: 806.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 7.74 (d, J=7.2 Hz, 1H), 7.66 (s, 1H), 7.60-7.29 (m, 5H), 7.10-6.96 (m, 1H), 6.83 (s, 2H), 4.07 (d, J=6.4 Hz, 2H), 3.88-3.57 (m, 14H) , 2.33-2.24 (m, 2H), 1.58-1.40 (m, 9H), 1.01-0.78 (m, 4H).

[001495] Step 8: Preparation of tert-butyl 4-(6-chloro-8-fluoro-2-((4-(((8)-13-((2S,4R)-4-hydroxy-2-(( (S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carbonyl)-14,14-dimethyl-11-oxo-3,6,9-trioxa-12 -azapentadecyl)oxy)phenyl)amino)-7-(3-hydroxynaphthalene-1-yl)quinazolin-4-yl)piperazine-1-carboxylate

[001496] To a solution of (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (94 mg, 0.21 mmol, 1 eq.) in N,N-dimethylformamide (3 ml) was added N,N-diisopropylethylamine (82 mg, 0 .63 mmol, 3 eq.) and 2-[2-[2-[2-[4-[[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-( 3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]phenoxy]ethoxy]ethoxy]ethoxy]acetic acid (170 mg, 0.21 µmol, 1 eq.), 1-hydroxybenzotriazole (34 mg, 0.25 mmol, 1.2 eq.), the reaction mixture was stirred at 15°C for 0.15 hour, then 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (49 mg, 0.25 mmol, 1.2 eq.) was added to the mixture. 2 eq.). The reaction mixture was stirred at 15°C for 0.15 hours. Then heated to 30°C for 2 hours. The reaction mixture was quenched by adding water (10 ml) and then diluted with water (30 ml), filtered and extracted with ethyl acetate (40 ml x 3). The combined organic layers were washed with brine (80 ml), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative TLC (dichloromethane:methanol 10:1). Desired compound tert-butyl 4-[6-chloro-8-fluoro-2-[4-[2-[2-[2-[2-[[(1S)-1-[(2S,4R)-4- hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl]amino]-2- oxo-ethoxy]ethoxy]ethoxy]ethoxy]anilino]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (180 mg, 0.14 mmol, 65% yield, purity 94 %) was obtained as a yellow solid. LC/MS (ESI) m/z: 617.2 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ ) δ 8.67 (d, J=4.4 Hz, 1H), 7.74 (dd, J=3.2, 8.4 Hz, 1H), 7, 69-7.60 (m, 3H), 7.46-7.29 (m, 6H), 7.25-7.09 (m, 2H), 6.92 (d, J=9.2 Hz, 2H), 5.06 (dd, J=6.8, 11.6 Hz, 1H), 4.88-4.75 (m, 1H), 4.59-4.41 (m, 2H), 4 .21-4.11 (m, 2H), 4.09-3.98 (m, 2H), 3.96-3.86 (m, 1H), 3.84-3.80 (m, 2H) , 3.77-3.55 (m, 16H), 2.49 (d, J=8.8 Hz, 3H), 2.10-1.97 (m, 1H), 1.52 (s, 9H ), 1.44 (d, J=6.8 Hz, 3H), 1.30-1.22 (m, 1H), 1.04 (d, J=4.4 Hz, 9H), 0.93 -0.79 (m, 1H).

[001497] Step 9: Preparation of (2S,4R)-1-((2S)-2-(tert-butyl)-14-(4-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene- 1-yl)-4-(piperazin-1-yl)quinazolin-2-yl)amino)phenoxy)-4-oxo-6,9,12-trioxa-3-azatetradecanoyl)-4-hydroxy-N-(( S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001498] To a solution of tert-butyl 4-[6-chloro-8-fluoro-2-[4-[2-[2-[2-[2-[[(1 S)-1-[(2S,4R )-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino ]-2-oxo-ethoxy]ethoxy]ethoxy]ethoxy]anilino]-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazine-1-carboxylate (180 mg, 0.15 mmol, 1 equiv .) in dichloromethane (2 ml) was added hydrogen chloride/dioxane (4 M, 2 ml, 55 eq.). The reaction mixture was stirred at 15°C for 0.15 hours. The reaction mixture was concentrated under reduced pressure to obtain a residue. Acetonitrile was added to the product, then concentrated under reduced pressure to obtain the product. Desired compound (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[4-[[6-chloro-8-fluoro-7-(3-hydroxy- 1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]phenoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N[( 1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (170 mg, HCl obtained as a yellow solid. LC/MS (ESI) m/z: 567 .3 [M/2+1] ⁺ .

[001499] Step 10: Preparation of (2S,4R)-1-((2S)-14-(4-((4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-( 3-hydroxynaphthalene-1-yl)quinazolin-2-yl)amino)phenoxy)-2-(tert-butyl)-4-oxo-6,9,12-trioxa-3-azatetradecanoyl)-4-hydroxy-N- ((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[001500] To a solution of (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[4-[[6-chloro-8-fluoro-7-(3 -hydroxy-1-naphthyl)-4-piperazin-1-yl-quinazolin-2-yl]amino]phenoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy- N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (70 mg, 0.06 mmol, 1 equiv, HCl) in dichloromethane (5 ml ) 2,6-lutidine (64 mg, 0.6 mmol, 10 eq.) was added. The mixture was stirred at 15°C for 0.15 hour, then prop-2-enoyl chloride (5 mg, 0.06 mmol, 1 eq.) was added to the mixture at minus 78°C. The reaction mixture was stirred at minus 78°C for 0.15 hours. The reaction mixture was quenched by adding water (10 ml) and then diluted with water (20 ml) and extracted with dichloromethane (20 ml x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC. Desired compound (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[4-[[6-chloro-8-fluoro-7-(3-hydroxy- 1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]amino]phenoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]- 4-Hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (20.3 mg, 0.016 mmol, 26% yield, 94% purity ) was obtained as a yellow solid. LC/MS (ESI) m/z: 593.9 [M/2+1] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 9.53 (s, 1H), 8.97 (s, 1H), 8.41 (d, J=7.6 Hz, 1H), 8, 32-8.25 (m, 1H), 7.92-7.68 (m, 4H), 7.49-7.31 (m, 6H), 7.30-7.19 (m, 3H), 7.08 (d, J=2.1 Hz, 1H), 6.92-6.78 (m, 3H), 6.24-6.10 (m, 1H), 5.80-5.70 ( m, 1H), 4.95-4.81 (m, 1H), 4.54 (d, J=9.6 Hz, 1H), 4.44 (t, J=8.0 Hz, 1H), 4.28 (s, 1H), 4.04 (t, J=3.6 Hz, 2H), 3.95 (s, 2H), 3.88-3.69 (m, 9H), 3.66 -3.51 (m, 11H), 2.44 (s, 3H), 2.10-1.98 (m, 1H), 1.82-1.71 (m, 1H), 1.44-1 .27 (m, 3H), 0.93 (s, 9H).

[001501] Example of the synthesis of (2S,4R)-N-((1R)-15-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1 -yl)quinazolin-2-yl)amino)-12-methyl-1-(4-(4-methylthiazol-5-yl)phenyl)-13-oxo-3,6,9-trioxa-12-azapentadecyl)- 4-Hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide (representative compound 487)

[001502] Step 1: Preparation of (2S,4R)-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)-N-((R)-13 -(4-(4-methylthiazol-5-yl)phenyl)-5,8,11-trioxa-2-azatridecan-13-yl)pyrrolidin-2-carboxamide

[001503] To a solution of tert-butyl N-[2-[2-[2-[(2R)-2-[[(2S,4R)-4-hydroxy-1-[(2R)-3-methyl-2 -(3-methylisoxazol-5-yl)butanoyl]pyrrolidin-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethoxy]ethoxy]ethoxy]ethyl]-N-methyl- carbamate (2S mg, 0.04 mmol, 1 eq.) in dichloromethane (1 ml), hydrochloric acid/dioxane (4 M, 1 ml, 108.28 eq.) was added, the mixture was stirred at 20°C for 0. 5 o'clock. The mixture was concentrated to obtain the product. Compound (2S,4R)-4-hydroxy-N-[(1R)-2-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]-1-[4-(4-methylthiazol-5 -yl)phenyl]ethyl]-1-[(2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide was obtained as a pale yellow oil. LC/MS (ESI) m/z: 658.2 [M+1] ⁺ .

[001504] Step 2: Preparation of (2S,4R)-N-((1R)-15-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene -1-yl)quinazolin-2-yl)amino)-12-methyl-1-(4-(4-methylthiazol-5-yl)phenyl)-13-oxo-3,6,9-trioxa-12-azapentadecyl )-4-hydroxy-1-((R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide

[001505] To a solution of (2S,4R)-4-hydroxy-N-[(1R)-2-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]-1-[4-(4 -methylthiazol-5-yl)phenyl]ethyl]-1-[(2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (26 mg, 0.04 mmol, 1 eq., hydrochloride), 3-[[4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino] propanoic acid (24 mg, 0.04 mmol, 1.2 eq.), 1-hydroxybenzotriazole (7 mg, 0.055 mmol, 1.5 eq.) and N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (11 mg , 0.055 mmol, 1.5 eq.) in N,N-dimethylformamide (1 ml) was added diisopropylethylamine (14 mg, 0.11 mmol, 3 eq.). The reaction solution was stirred at 20°C for 15 hours. The residue was purified by preparative HPLC, the acetonitrile fraction was removed, and the residue was lyophilized to give the product. (2S,4R)-N-[(1R)-2-[2-[2-[2-[3-[[4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7 -(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethoxy]-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]- 4-hydroxy-1-[(2R)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (6.5 mg, 0.005 mmol, yield 14.7%, purity 97 .9%) was obtained as a white solid. LC/MS (ESI) m/z: 1177.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 8.90-8.79 (m, 1H), 7.79 (s, 1H), 7.72 (d, J=8.3 Hz, 1H) , 7.41-7.29 (m, 5H), 7.25-7.11 (m, 3H), 7.03-6.98 (m, 1H), 6.22-6.16 (m, 1H), 5.06-4.97 (m, 1H), 4.71-4.55 (m, 3H), 4.45-4.29 (m, 1H), 3.81 (br dd, J =8.7, 19.6 Hz, 12H), 3.62-3.48 (m, 12H), 3.14-3.07 (m, 2H), 2.91 (br s, 4H), 2 .48-2.33 (m, 4H), 2.24-2.14 (m, 7H), 1.99-1.86 (m, 1H), 1.06-0.91 (m, 3H) , 0.87-0.74 (m, 3H).

[001506] Example of the synthesis of (2S,4R)-N-((1R)-15-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1 -yl)quinazolin-2-yl)amino)-12-methyl-1-(4-(4-methylthiazol-5-yl)phenyl)-13-oxo-3,6,9-trioxa-12-azapentadecyl)- 4-Hydroxy-1-((S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide (typical compound 488)

[001507] Step 1: Preparation of (2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)-N-((R)-13 -(4-(4-methylthiazol-5-yl)phenyl)-5,8,11-trioxa-2-azatridecan-13-yl)pyrrolidin-2-carboxamide

[001508] To a solution of tert-butyl N-[2-[2-[2-[(2R)-2-[[(2S,4R)-4-hydroxy-1-[(2S)-3-methyl-2 -(3-methylisoxazol-5-yl)butanoyl]pyrrolidin-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethoxy]ethoxy]ethoxy]ethyl]-N-methyl- carbamate (30 mg, 0.04 mmol, 1 eq.) in dichloromethane (1 ml), hydrochloric acid/dioxane (4 M, 1 ml, 101.06 eq.) was added, the mixture was stirred at 20°C for 0. 5 o'clock. The mixture was concentrated to obtain the product. (2S,4R)-4-hydroxy-N-[(1R)-2-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]-1-[(2S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide was obtained as a pale yellow oil. LC/MS (ESI) m/z: 658.2 [M+1] ⁺ .

[001509] Step 2: Preparation of (2S,4R)-N-((1R)-15-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene -1-yl)quinazolin-2-yl)amino)-12-methyl-1-(4-(4-methylthiazol-5-yl)phenyl)-13-oxo-3,6,9-trioxa-12-azapentadecyl )-4-hydroxy-1-((S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl)pyrrolidine-2-carboxamide

[001510] To a solution of (2S,4R)-4-hydroxy-N-[(1R)-2-[2-[2-[2-(methylamino)ethoxy]ethoxy]ethoxy]-1-[4-(4 -methylthiazol-5-yl)phenyl]ethyl]-1-[(2S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (27 mg, 0.04 mmol, 1 eq., hydrochloride), 3-[[4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino] propanoic acid (25 mg, 0.046 mmol, 1.2 eq.), 1-hydroxybenzotriazole (8 mg, 0.058 mmol, 1.5 eq.) and N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (11 mg, 58 .33 µmol, 1.5 eq.) in N,N-dimethylformamide (1 ml) was added diisopropylethylamine (15 mg, 0.12 mmol, 3 eq.). The reaction solution was stirred at 20°C for 15 hours. The residue was purified by preparative HPLC, the acetonitrile fraction was removed, and the residue was lyophilized to give the product. (2S,4R)-N-[(1R)-2-[2-[2-[2-[3-[[4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7 -(3-hydroxy-1-naphthyl)quinazolin-2-yl]amino]propanoyl-methyl-amino]ethoxy]ethoxy]ethoxy]-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]- 4-hydroxy-1-[(2S)-3-methyl-2-(3-methylisoxazol-5-yl)butanoyl]pyrrolidine-2-carboxamide (7.9 mg, 0.006 mmol, 17% yield, purity 98.7 %) was obtained as a white solid. LC/MS (ESI) m/z: 1177.4 [M+1] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 8.84 (d, J=1.2 Hz, 1H), 7.80 (br s, 1H), 7.73 (br d, J=8, 6 Hz, 1H), 7.42-7.33 (m, 5H), 7.25-7.13 (m, 3H), 7.03-6.98 (m, 1H), 6.21-6 ,18 (m, 1H), 5.06 (br s, 1H), 4.69-4.52 (m, 3H), 4.44-4.33 (m, 1H), 3.84-3, 71 (m, 12H), 3.62-3.49 (m, 12H), 3.16-3.07 (m, 2H), 2.94-2.70 (m, 4H), 2.45- 2.37 (m, 4H), 2.23-2.14 (m, 7H), 2.01-1.88 (m, 1H), 1.04 (d, J=6.5 Hz, 3H) , 0.84 (br d, J=6.8 Hz, 3H).

[001511] Synthesis example of (2S,4R)-1-((2S)-2-(3-(4-(2-((4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro -7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)(3-(dimethylamino)-3-oxopropyl)amino)ethoxy)butoxy)isoxazol-5-yl)-3-methylbutanoyl)-4-hydroxy -N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (typical compound 495)

[001512] Step 1: Preparation of ((2-(4-bromobutoxy)ethoxy)methyl)benzene

[001513] Sodium hydroxide (42.05 g, 1.05 mol, 8 eq.) and tetrabutylammonium hydrogen sulfate (44.62 g, 131.41 mmol, 1 eq.), 1,4-dibromobutane (42.56 g, 197.11 mmol, 23.78 ml, 1.5 eq. .). The mixture was stirred at 20°C for 16 hours. Hydrochloric acid (1 N) was added to acidify the aqueous phase (PH=4) and the solution was extracted with ethyl acetate (100 ml x 4). The combined organic layers were dried, filtered and concentrated to leave a residue. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 50/1 to 20/1). The compound 2-(4-bromobutoxy)ethoxymethylbenzene (20 g, 69.64 mmol, 52% yield) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 1.70-1.80 (m, 2H), 1.92-2.01 (m, 2H), 3.45 (t, J=6.8 Hz , 2H), 3.52 (t, J=6.4 Hz, 2H), 3.61-3.65 (m, 4H), 4.59 (s, 2H), 7.26-7.32 ( m, 1H), 7.36 (d, J=4.8 Hz, 4H).

[001514] Step 2: Preparation of methyl 2-(3-(4-(2-(benzyloxy)ethoxy)butoxy)isoxazol-5-yl)-3-methylbutanoate

[001515] A mixture of methyl 2-(3-hydroxyisoxazol-5-yl)-3-methyl-butanoate (3 g, 15.06 mmol, 1 eq.), 2-(4-bromobutoxy)ethoxymethylbenzene (4.33 g, 15.06 mmol, 1 eq.), potassium carbonate (4.16 g, 30.12 mmol, 2 eq.) in N,N-dimethylformamide (40 ml) was degassed and purged with nitrogen (3 times) and then the mixture was stirred at a temperature of 80°C for 2 hours in a nitrogen atmosphere. The residue was diluted with water (200 ml) and extracted with ethyl acetate (50 ml x 3). The combined organic layers were washed with brine (50 ml) and dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 10/1 to 1/1). The compound methyl 2-[3-[4-(2-benzyloxyethoxy)butoxy]isoxazol-5-yl]-3-methyl-butanoate (4.2 g, 10.36 mmol, 68% yield) was obtained as a colorless oil. ¹ H-NMR (400 MHz, CD ₃ OD) δ 0.91 (d, J=6.4 Hz, 3H), 0.99 (d, J=6.4 Hz, 3H), 1.67-1 .77 (m, 2H), 1.81-1.90 (m, 2H), 2.35 (dquin, J=8.4, 6.4, 6.4, 6.4, 6.4 Hz, 1H), 3.50-3.59 (m, 3H), 3.60-3.65 (m, 4H), 3.71 (s, 3H), 4.21 (t, J=6.4 Hz, 2H), 4.55 (s, 2H), 6.00 (s, 1H), 7.24-7.31 (m, 1H), 7.32-7.37 (m, 3H).

[001516] Step 3: Preparation of methyl 2-(3-(4-(2-hydroxyethoxy)butoxy)isoxazol-5-yl)-3-methylbutanoate

[001517] To a solution of methyl 2-[3-[4-(2-benzyloxyethoxy)butoxy]isoxazol-5-yl]-3-methyl-butanoate (4.2 g, 10.36 mmol, 1 eq.) in 1 ,2-dichloroethane (80 ml), water (8 ml) was added dichlorodicyanobenzoquinone (11.76 g, 51.79 mmol, 5 eq.). The mixture was stirred at 20°C for 1 hour. The reaction mixture was quenched by adding saturated sodium bicarbonate solution (150 ml) at 25°C and then diluted with water (50 ml). The organic phase was separated and extracted with ethyl acetate (50 ml x 4). The combined organic layers were washed with brine (20 ml), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 10/1 to 1/1). The compound methyl 2-[3-[4-(2-hydroxyethoxy)butoxy]isoxazol-5-yl]-3-methyl-butanoate (2.7 g, 8.56 mmol, 82% yield) was obtained as a purple liquid. ¹ H-NMR (400 MHz, CD ₃ OD) δ 0.85-0.95 (m, 3H), 0.95-1.04 (m, 3H), 1.66-1.78 (m, 2H ), 1.79-1.92 (m, 2H), 2.25-2.43 (m, 1H), 3.46-3.60 (m, 5H), 3.65 (br d, J= 3.6 Hz, 2H), 3.72 (br d, J=1.2 Hz, 3H), 4.12-4.28 (m, 2H), 6.00 (s, 1H).

[001518] Step 4: Preparation of methyl 2-(3-(4-(2-bromoethoxy)butoxy)isoxazol-5-yl)-3-methylbutanoate

[001519] To a solution of methyl 2-[3-[4-(2-hydroxyethoxy)butoxy]isoxazol-5-yl]-3-methyl-butanoate (2.7 g, 8.56 mmol, 1 eq.) in tetrahydrofuran (60 ml) carbon tetrabromide (8.52 g, 25.68 mmol, 3 eq.) was added, and then triphenylphosphine (6.74 g, 25.68 mmol, 3 eq.) in tetrahydrofuran (20 ml) was added dropwise at 0°C. The resulting mixture was stirred at a temperature of 25°C for 10 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 20/1 to 5/1). The compound methyl 2-[3-[4-(2-bromoethoxy)butoxy]isoxazol-5-yl]-3-methyl-butanoate (3.1 g, 8.20 mmol, 95% yield) was obtained as a yellow oil. ¹ H-NMR (400 MHz, CDCl ₃ ) δ 0.93 (d, J=6.8 Hz, 3H), 1.00 (d, J=6.4 Hz, 3H), 1.70-1, 78 (m, 2H), 1.83-1.91 (m, 2H), 2.30-2.40 (m, 1H), 3.44-3.50 (m, 3H), 3.55 ( t, J=6.4 Hz, 2H), 3.73 (s, 3H), 3.73-3.78 (m, 2H), 4.24 (t, J=6.4 Hz, 2H), 5.87(s, 1H).

[001520] Step 5: Preparation of methyl 2-(3-(4-(2-((tert-butoxycarbonyl)(3-(dimethylamino)-3-oxopropyl)amino)ethoxy)butoxy)isoxazol-5-yl)-3 -methylbutanoate

[001521] To a solution of methyl 2-[3-[4-(2-bromoethoxy)butoxy]isoxazol-5-yl]-3-methyl-butanoate (1.5 g, 3.97 mmol, 1 eq.) and 3 -amino-N,N-dimethylpropanamide (750 mg, 4.91 mmol, 1.24 eq., hydrochloride) in ethyl alcohol (40 ml) was added N,N-diisopropylethylamine (2.60 g, 20.12 mmol , 3.50 ml, 5.07 eq.). The resulting mixture was stirred at 80°C for 12 hours. The mixture was concentrated to obtain a residue. The compound methyl 2-[3-[4-[2-[[3-(dimethylamino)-3-oxo-propyl]amino]ethoxy]butoxy]isoxazol-5-yl]-3-methyl-butanoate (1 g) was prepared in the form of a yellow oil. LC/MS (ESI) m/z: 414.3 [M+1] ⁺ . To a solution of methyl 2-[3-[4-[2-[[3-(dimethylamino)-3-oxo-propyl]amino]ethoxy]butoxy]isoxazol-5-yl]-3-methyl-butanoate (1 g, di-tert-butyl dicarbonate (1 .06 g, 4.84 mmol, 1.11 ml, 2 eq.) at 0°C. The reaction mixture was stirred at 25°C for 2 hours. The mixture was poured into ice water (60 ml) and stirred for 5 minutes. The aqueous phase was extracted with dichloromethane (40 ml x 3). The combined organic phase was washed with brine (30 ml), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative HPLC. The compound is methyl 2-[3-[4-[2-[tert-butoxycarbonyl-[3-(dimethylamino)-3-oxo-propyl]amino]ethoxy]butoxy]isoxazol-5-yl]-3-methyl-butanoate ( 0.51 g, 992.95 µmol, 41% yield) was obtained as a colorless oil. LC/MS (ESI) m/z: 514.2 [M+1] ⁺ .

[001522] Step 6: Preparation of 2-(3-(4-(2-((tert-butoxycarbonyl)(3-(dimethylamino)-3-oxopropyl)amino)ethoxy)butoxy)isoxazol-5-yl)-3- methylbutanoic acid

[001523] To a solution of methyl 2-[3-[4-[2-[tert-butoxycarbonyl-[3-(dimethylamino)-3-oxo-propyl]amino]ethoxy]butoxy]isoxazol-5-yl]-3- methyl butanoate (450 mg, 876.13 µmol, 1 eq.) in tetrahydrofuran (8 ml), and methyl alcohol (8 ml), and water (8 ml) was added lithium hydroxide monohydrate (110.30 mg, 2.63 mmol, 3 eq.). The resulting mixture was stirred at 25°C for 12 hours. The reaction mixture was adjusted to pH=5 with dilute hydrochloric acid (1 N) at 0°C and extracted with ethyl acetate (30 ml x 3). The combined organic layers were washed with brine (10 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated to give the product. Compound 2-[3-[4-[2-[tert-butoxycarbonyl-[3-(dimethylamino)-3-oxo-propyl]amino]ethoxy]butoxy]isoxazol-5-yl]-3-methyl-butanoic acid ( 450 mg) was obtained as a colorless oil. LC/MS (ESI) m/z: 500.3 [M+1] ⁺ .

[001524] Step 7: Preparation of tert-butyl (3-(dimethylamino)-3-oxopropyl)(2-(4-((5-((S)-1-((2S,4R)-4-hydroxy-2 -((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)oxy)butoxy)ethyl)carbamate

[001525] To a solution of 2-[3-[4-[2-[tert-butoxycarbonyl-[3-(dimethylamino)-3-oxo-propyl]amino]ethoxy]butoxy]isoxazol-5-yl]-3-methyl -butanoic acid (400 mg, 800.65 µmol, 1 eq.) and [dimethylamino(triazolo[4,5-b]pyridin-3-yloxy)methylene]-dimethyl-ammonium; hexafluorophosphate (921 mg, 2.42 mmol, 3.03 eq.) in N,N-dimethylformamide (8 ml) was added (2S,4R)-4-hydroxy-N-[[4-(4-methylthiazol-5- yl)phenyl]methyl]pyrrolidine-2-carboxamide (255 mg, 803.39 µmol, 1 eq.) and N,N-diisopropylethylamine (420 mg, 3.25 mmol, 566.04 µl, 4.06 eq.) . The resulting mixture was stirred at 25°C for 12 hours. The mixture was poured into ice water (40 ml) and stirred for 5 minutes. The aqueous phase was extracted with ethyl acetate (30 ml x 3). The combined organic phase was washed with brine (15 ml x 2), dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography (dichloromethane/methyl alcohol 20/1 to 10/1). Tert-butyl compound N-[3-(dimethylamino)-3-oxo-propyl]-N-[2-[4-[5-[1-[(2S,4R)-4-hydroxy-2-[[4 -(4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidine-1-carbonyl]-2-methyl-propyl]isoxazol-3-yl]oxybutoxy]ethyl]carbamate (650 mg, 748.45 µmol, 93% yield , purity 92%) was obtained as an off-white solid. LC/MS (ESI) m/z: 199 A [M+1] ⁺ .

[001526] Step 8: Preparation of tert-butyl (3-(dimethylamino)-3-oxopropyl)(2-(4-((5-((S)-1-((2S,4R)-4-hydroxy-2 -((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)isoxazol-3-yl)oxy)butoxy)ethyl)carbamate

[001527] Tert-butyl N-[3-(dimethylamino)-3-oxo-propyl]-N-[2-[4-[5-[1-[(2S)-4-hydroxy-2-[[4 -(4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidine-1-carbonyl]-2-methyl-propyl]isoxazol-3-yl]oxybutoxy]ethyl]carbamate (630.00 mg, 788.50 µmol, 1 eq.) were separated using SFC. The reaction mixture was concentrated under reduced pressure to obtain a residue. Tert-butyl compound N-[3-(dimethylamino)-3-oxo-propyl]-N-[2-[4-[5-[(1S)-1-[(2S,4R)-4-hydroxy-2 -[[4-(4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidine-1-carbonyl]-2-methyl-propyl]isoxazol-3-yl]oxybutoxy]ethyl]carbamate (0.3 g, 364, 21 µmol, yield 46%, purity 97%) was obtained as a light yellow solid.

[001528] Step 9: Preparation of (2S,4R)-1-((S)-2-(3-(4-(2-((3-(dimethylamino)-3-oxopropyl)amino)ethoxy)butoxy)isoxazole -5-yl)-3-methylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[001529] A mixture of tert-butyl N-[3-(dimethylamino)-3-oxo-propyl]-N-[2-[4-[5-[(1S)-1-[(2S,4R)-4- Hydroxy-2-[[4-(4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidine-1-carbonyl]-2-methyl-propyl]isoxazol-3-yl]oxybutoxy]ethyl]carbamate (150 mg, 187 .74 µmol, 1 eq.) and hydrochloric acid/dioxane (4 M, 10 ml) were stirred at 25°C for 3 hours. The mixture was concentrated to obtain the product. Compound (2S,4R)-1-[(2S)-2-[3-[4-[2-[[3-(dimethylamino)-3-oxo-propyl]amino]ethoxy]butoxy]isoxazol-5-yl ]-3-methyl-butanoyl]-4-hydroxy-N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (130 mg, 176.79 µmol, 94% yield, hydrochloride) was obtained as a light yellow solid. LC/MS (ESI) m/z: 699.3 [M+1] ⁺ .

[001530] Step 10: Preparation of (2S,4R)-1-((2S)-2-(3-(4-(2-((4-(4-acetylpiperazin-1-yl)-6-chloro-8 -fluoro-7-(3-hydroxynaphthalene-1-yl)quinazolin-2-yl)(3-(dimethylamino)-3-oxopropyl)amino)ethoxy)butoxy)isoxazol-5-yl)-3-methylbutanoyl)-4 -hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[001531] To a solution of 1-[4-[2,6-dichloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazin-1-yl]ethanone (40 mg, 82 .42 µmol, 1 eq.) and (2S,4R)-1-[(2S)-2-[3-[4-[2-[[3-(dimethylamino)-3-oxo-propyl]amino]ethoxy ]butoxy]isoxazol-5-yl]-3-methyl-butanoyl]-4-hydroxy-N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (130 mg, 176 .79 µmol, 2.15 equiv., hydrochloride) in dioxane (1 ml) was added N,N-diisopropylethylamine (159.78 mg, 1.24 mmol, 215.33 µl, 15 equiv.). The mixture was stirred at 100°C for 10 hours. The mixture was concentrated to obtain a residue. The residue was purified by semipreparative reverse phase HPLC. Compound (2S,4R)-1-[(2S)-2-[3-[4-[2-[[4-(4-acetylpiperazin-1-yl)-6-chloro-8-fluoro-7-( 3-hydroxy-1-naphthyl)quinazolin-2-yl]-[3-(dimethylamino)-3-oxo-propyl]amino]ethoxy]butoxy]isoxazol-5-yl]-3-methyl-butanoyl]-4- Hydroxy-N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (29.1 mg, 24.09 µmol, 29% yield, 95% purity) was obtained as a yellow solid substances. LC/MS (ESI) m/z: 1169.5 [M+23] ⁺ ; ¹ H-NMR (400 MHz, CD ₃ OD) δ 0.57-0.65 (m, 1H), 0.76 (d, J=6.4 Hz, 1H), 0.85-0.92 (m, 2H), 1.03 (d, J=6.4 Hz, 2H), 1.54-1.79 (m, 4 H), 2.01-2.10 (m, 1H), 2.14 ( s, 3H), 2.19-2.27 (m, 1H), 2.27-2.38 (m, 1H), 2.40-2.47 (m, 3H), 2.76-2, 93 (m, 5H), 2.99-3.16 (m, 3H), 3.42-3.49 (m, 2H), 3.63-3.81 (m, 12H,) 3.84- 4.15 (m, 6H), 4.34-4.42 (m, 2H), 4.47 (br s, 1H), 4.54-4.69 (m, 2H), 5.84-5 .98 (m, 1H), 6.97-7.05 (m, 1H), 7.10-7.19 (m, 1H), 7.23 (br d, J=2.4 Hz, 2H) , 7.30-7.40 (m, 4H), 7.41-7.50 (m, 1H), 7.72 (d, J=8.4 Hz, 1H), 7.75-7.80 (m, 1H), 8.79-8.89 (m, 1H).

Protein level control

[001532] This disclosure also provides methods for monitoring protein levels in a cell. This is based on the use of compounds described herein that are known to interact with a particular target protein in such a way that degradation of the target protein in vivo will result in control of the amount of protein in the biological system, preferably with some therapeutic benefit.

[001533] The following examples are used to assist in the description of the present invention, but they should not be construed as limiting the proposed invention in any way.

Specific embodiments of the Proposed Invention

[001534] The present invention includes the following specific embodiments. These further embodiments may include all of the properties set forth in the following embodiment, as provided. Where applicable, subsequent embodiments may also include features set forth in any of the following embodiments, inclusive or alternative (e.g., the eighth embodiment may include features set forth in the first embodiment as set forth and/or features of any of second to seventh embodiment).

[001535] In certain embodiments, the disclosure provides the following exemplary bifunctional KRas molecules (exemplary compounds 1-10 of Tables 4 and 5 and 11-249, 254-454 and 458-573 of Tables 6, 8, 10 and 12), including salts thereof , prodrugs, polymorphs, analogs, derivatives and deuterated forms.

[001536] In any aspect or embodiment herein, the bifunctional compound of the invention comprises a PTM from Table 1 (e.g., PTM-1, PTM-2, PTM-3, PTM-4, PTM-5, or PTM- 6), linker from Table 2 (for example, L-1, L-2, L-3, L-4, L-5 or L-6) and ULM from Table 3 (for example, ULM-1, ULM-2, ULM-3, ULM-4 or ULM-5), including their salts, prodrugs, polymorphs, analogs, derivatives and deuterated forms:

[001537] Table 1. Typical PTMs of typical PROTACs according to the present invention

[001538] Table 2. Typical linkers of typical PROTACs as proposed

invention

[001539] Table 3. Typical ULMs of typical PROTACs according to the present invention

[001540] In certain exemplary embodiments as described herein, a compound is provided having a linker selected from Table 2 (e.g., L-1, L-2, L-3, L-4, L-5, or L- 6) associated with the RTM from Table 1 (for example, RTM-1, RTM-2, RTM-3, RTM-4, RTM-5 or RTM-6) and the ULM from Table 3 (for example, ULM-1, ULM- 2, ULM-3, ULM-4 or ULM-5). For example, the PTM of table 1, the linker of table 2 and the ULM of table 3 can be combined in any desired combination, for example, PTM-1/L-1/ULM-1 or PTM-2/L-1/ULM-1, etc. ., thereby forming typical compounds of the present invention.

[001541] In one aspect, the present invention provides a bifunctional compound having the chemical structure:

ULM-L-RTM,

or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate, polymorph or prodrug thereof, wherein: the ULM is a small molecule E3 ubiquitin ligase binding fragment that binds E3 ubiquitin ligase; PTM is a small molecule containing a moiety targeting the Kirsten rat sarcoma protein (KRas); and L represents a bond or chemically linked moiety connecting the ULM and the PTM.

[001542] In any aspect or embodiment described herein, the E3 ubiquitin ligase binding moiety that targets the E3 ubiquitin ligase is selected from the group consisting of von Hippel-Lindau (VLM), cereblon (CLM), mouse double minute homolog2 (MLM) and IAP (ILM).

[001543] In any aspect or embodiment described herein, the PTM is represented as:

Where:

is aryl, heteroaryl, cycloalkyl or heterocycloalkyl; X _PTM is C or N;

W _PTM is selected from the group consisting of optionally substituted C3-C6 cycloalkyl and optionally substituted C3-C6 heteroalkyl, optionally substituted C3-C6 heterocycloalkyl, optionally substituted aryl (e.g., optionally substituted C5-C7 aryl), optionally substituted heteroaryl (e.g., optionally substituted C5-C7 heteroaryl);

R _PTM1A is NR _PTM9 R _PTM10 , OR _PTM9 R _PTM10 , H, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted C3-C6 cycloalkyl, optionally substituted O-(C3-C6 cycloalkyl), optionally substituted-O- _{C1- 4} alkyl-C _3-6 cycloalkyl, optionally substituted C3-C6 heteroalkyl, optionally substituted O-(C3-C6 heteroalkyl), optionally substituted O-C _1-4 alkyl-C _3-6 heteroalkyl, optionally substituted O-C _{1- 4} alkyl-C _3-6 heterocycloalkyl, optionally substituted aryl (e.g., optionally substituted C5-C7 aryl), optionally substituted O-aryl (e.g., optionally substituted O-(C5-C7 aryl)), optionally substituted heteroaryl (e.g., optionally substituted C5-C7 heteroaryl), optionally substituted O-heteroaryl (e.g. optionally substituted O-(C5-C7 heteroaryl)), optionally substituted (e.g., optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted (e.g., optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), optionally substituted (e.g., optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), optionally substituted (e.g., optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), optionally substituted (e.g., optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), optionally substituted (e.g., optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), optionally substituted (eg, optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), where N* represents the N atom of a heterocycloalkyl linker (L);

R _PTM1B is NR _PTM9 R _PTM10 , OR _PTM9 R _PTM10 , H, optionally substituted alkyl, optionally substituted O-alkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted O-(C3-C6 cycloalkyl), optionally substituted-O-C _1-4 alkyl-C _3-6 cycloalkyl, optionally substituted C3-C6 heteroalkyl, optionally substituted O-(C3-C6 heteroalkyl), optionally substituted OC _1-4 alkyl-C _3-6 heteroalkyl, optionally substituted aryl (e.g., optionally substituted C5-C7 aryl), optionally substituted O-aryl (e.g., optionally substituted O-(C5-C7 aryl)), optionally substituted heteroaryl (e.g., optionally substituted C5-C7 heteroaryl), optionally substituted O-heteroaryl (e.g., optionally substituted O(C5-C7 heteroaryl)), optionally substituted (e.g., optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl), optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted optionally substituted

R _PTM9 and R _PTM10 are each independently H, optionally substituted C1-C6 alkyl, optionally substituted aliphatic amine, optionally substituted aliphatic amide;

R _PTM2 is H, (C=O)R _PTM2 ^' ; optionally substituted straight or branched alkyl;

R _PTM2 ^' represents an optionally substituted linear or branched alkyl, an optionally substituted alkene, -N (R _PTM8 ) ₂ or -C(OH) ₂ ;

R _PTM3 is alkyl, alkoxy, phenyl or naphthalene, each independently substituted with OH, H, halogen;

R _PTM4A is OH, H, halogen, optionally substituted linear or branched C1-C6 alkyl;

R _PTM4B represents OH, H, halogen, optionally substituted linear or branched C1-C6 alkyl;

R _PTM5 is selected from the group consisting of optionally substituted aryl, optionally substituted biaryl, optionally substituted heteroaryl, optionally substituted biheteroaryl, optionally substituted C ₃ -C ₆ cycloalkyl, optionally substituted C ₃ -C ₆ cycloheteroalkyl, halogen, H, optionally substituted linear or branched alkyl (eg, optionally substituted linear or branched C ₁ -C ₆ alkyl), OH and alkoxy;

R _PTM8 is H or alkyl (eg, C1 alkyl, C2 alkyl, C3 alkyl or C4 alkyl);

t represents 0, 1, 2, 3, 4, 5, 6; And

denotes the site of attachment of at least one of: linker, ULM, ULM', CLM, CLM', VLM, VLM', ILM, ILM', MLM, MLM', or a combination thereof.

[001544] In any aspect or embodiment described herein, the PTM is represented as:

Where:

X _PTM1 is NH or O;

R _PTM6 is aryl, heteroaryl, where N* represents the N atom of a heterocycloalkyl (eg C4-C8 heterocycloalkyl) linker (L);

R _PTM7 is H, aryl, O-aryl, heteroaryl, O-heteroaryl,

R _PTM9 is H, optionally substituted C1-C6 alkyl, optionally substituted aliphatic amine, optionally substituted aliphatic amide, optionally substituted (eg, optionally substituted with at least one alkyl, such that * carbon may be optionally substituted with alkyl);

may be a single bond or a double bond; And

denotes the site of attachment of at least one of: linker, ULM, ULM', CLM, CLM', VLM, VLM', ILM, ILM', MLM, MLM', or a combination thereof.

[001545] In any aspect or embodiment described herein, PTM is represented as the chemical structure:

[001546] In any aspect or embodiment described herein, the ULM is a von Hippel-Lindau ligase (VHL) binding moiety (VLM) with the chemical structure represented by:

Where:

X ¹ , X ² are each independently selected from the bond group, O, NR ^Y3 , CR ^Y3 R ^Y4 , C=O, C=S, SO and SO ₂ ;

R ^Y3 , R ^Y4 are each independently selected from the group H, linear or branched C _1-6 alkyl, optionally substituted with 1 or more halo, C _1-6 alkoxy, optionally substituted with 0-3 R ^P groups;

R ^P represents 0,1,2 or 3 groups, each independently selected from the group H, halo, -OH, C _1-3 alkyl, C=O;

W ³ is selected from the group of optionally substituted T, optionally substituted -TN(R ^1a R ^1b )X ³ , optionally substituted -TN(R ^1a R ^1b ), optionally substituted -T-aryl, optionally substituted -T-heteroaryl, optionally substituted - T-heterocyclyl, optionally substituted -T-biheterocyclyl, optionally substituted -NR ¹ -T-aryl, optionally substituted -NR ¹ -T-heteroaryl or optionally substituted -NR ¹ -T- heterocyclyl;

X ³ represents C=O, R ¹ , R ^1a , R ^1b ;

each of R ¹ , R ^1a , R ^1b independently selected from the group consisting of an H, straight or branched C ₁ -C ₆ alkyl group, optionally substituted with 1 or more halogens or -OH groups, R ^Y3 C=O, R ^Y3 C=S, R ^Y3 SO, R ^Y3 SO ₂ , N(R ^Y3 R ^Y4 )C=O, N(R ^Y3 R ^Y4 )C=S, N(R ^Y3 R ^Y4 )SO and N(R ^Y3 R ^Y4 )SO ₂ ; T is selected from the group of optionally substituted alkyl, -(CH ₂ ) _n - group, where each of the methylene groups is optionally substituted by one or two substituents selected from the group of halogen, methyl, optionally substituted alkoxy, linear or branched C ₁ -C ₆ alkyl groups optionally substituted with 1 or more halogens, C(O)NR ¹ R ^1a , or NR ¹ R ^1a , or R ¹ and R ^1a are linked to form an optionally substituted heterocycle, or -OH groups, or an optionally substituted amino acid side chain; And

n represents 0-6,

W ⁴ represents or

R _14a , R _14b are each independently selected from the group H, haloalkyl or optionally substituted alkyl;

W ⁵ is selected from the group of optionally substituted phenyl or optionally substituted 5-10 membered heteroaryl (e.g., optionally substituted with one or more [such as 1, 2, 3, 4 or 5] halogens, CN, optionally substituted alkyl, optionally substituted haloalkyl, optional substituted alkoxy, hydroxy or optionally substituted haloalkoxy),

R ₁₅ is selected from the group H, halogen, CN, OH, NO ₂ , NR _14a R _14b , OR _14a , CONR _14a R _14b , NR _14a COR _14b , SO ₂ NR _14a R _14b , NR _14a SO ₂ R _14b , optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl or optionally substituted cycloheteroalkyl;

and wherein the dotted line represents the attachment site of at least one PTM, another ULM (ULM'), or a chemical linker moiety associated with at least one PTM or ULM', or both ULMs.

[001547] In any aspect or embodiment described herein, the ULM is a von Hippel-Lindau ligase (VHL) binding moiety (VLM) with the chemical structure represented by:

Where:

W ³ is selected from the group of optionally substituted aryl, optionally substituted heteroaryl or

R ₉ and R ₁₀ independently represent hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted hydroxyalkyl, optionally substituted heteroaryl or haloalkyl, or R ₉ , R ₁₀ and the carbon atom to which they are attached form optionally substituted cycloalkyl;

R ₁₁ is selected from the group of optionally substituted heterocyclyl, optionally substituted alkoxy, optionally substituted heteroaryl, optionally substituted aryl, or

R ₁₂ is selected from the group H or optionally substituted alkyl;

R ₁₃ is selected from the group H, optionally substituted alkyl, optionally substituted alkylcarbonyl, optionally substituted (cycloalkyl)alkylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted arylcarbonyl, optionally substituted (heterocyclyl)carbonyl, or optionally substituted aralkyl;

R _14a , R _14b are each independently selected from the group H, haloalkyl or optionally substituted alkyl;

W ⁵ is selected from the group of optionally substituted phenyl or optionally substituted 5-10 membered heteroaryl (e.g., optionally substituted with one or more [such as 1, 2, 3, 4 or 5] halogens, CN, optionally substituted alkyl, optionally substituted haloalkyl, optional substituted alkoxy, hydroxy or optionally substituted haloalkoxy);

R ₁₅ is selected from the group H, halogen, CN, OH, NO ₂ , NR _14a R _14b , OR _14a , CONR _14a R _14b , NR _14a COR _14b , SO ₂ NR _14a R _14b , NR _14a SO ₂ R _14b , optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl or optionally substituted cycloheteroalkyl;

each R ₁₆ is independently selected from the group halogen, CN, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkoxy, hydroxy or optionally substituted haloalkoxy;

o represents 0, 1, 2, 3 or 4;

R ₁₈ is independently selected from the group of halogen, optionally substituted alkoxy, cyano, optionally substituted alkyl, haloalkyl, haloalkoxy or linker; And

p is 0, 1, 2, 3 or 4 and wherein the dotted line represents the attachment site of at least one PTM, another ULM (ULM'), or a chemical linker moiety bound to at least one PTM or ULM', or both ULM.

[001548] In any aspect or embodiment described herein, the ULM has a chemical structure selected from the group:

And

Where:

R ₁ is H, ethyl, isopropyl, t-butyl, sec-butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; optionally substituted alkyl, optionally substituted hydroxyalkyl, optionally substituted heteroaryl or haloalkyl;

R _14a is H, haloalkyl, optionally substituted alkyl, methyl, fluoromethyl, hydroxymethyl, ethyl, isopropyl or cyclopropyl;

R ₁₅ is selected from the group consisting of H, halogen, CN, OH, NO ₂ , optionally substituted heteroaryl, optionally substituted aryl, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy, optionally substituted cycloalkyl, or optionally substituted cycloheteroalkyl;

X represents C, CH ₂ or C=O

R ₃ is absent or is an optionally substituted 5 or 6 membered heteroaryl; And

wherein the dotted line represents the attachment site of at least one PTM, another ULM (ULM'), or a chemical linker moiety bound to at least one PTM or ULM', or both ULMs.

[001549] In any aspect or embodiment described herein, the ULM contains a group in accordance with the chemical structure:

ULM-g

Where:

R ^1' in ULM-g is an optionally substituted C ₁ -C ₆ alkyl group, optionally substituted -(CH ₂ ) _n OH, optionally substituted -(CH ₂ ) _n SH, optionally substituted (CH ₂ ) _n -O-( a C ₁ -C ₆ )alkyl group, an optionally substituted (CH ₂ ) _n -WCOCW-(C ₀ -C ₆ )alkyl group containing a WCOCW epoxy moiety, where each W independently represents an H or a C ₁ -C ₃ alkyl group, optionally substituted -(CH ₂ ) _n COOH, optionally substituted -(CH ₂ ) _n C(O)-(C ₁ -C ₆ alkyl), optionally substituted -(CH ₂ ) _n NHC(O)-R ₁ , optionally substituted -(CH ₂ ) _n C(O)-NR ₁ R ₂ , optionally substituted -(CH ₂ ) _n OC(O)-NR ₁ R ₂ , -(CH ₂ O) _n H, optionally substituted -(CH ₂ ) _n OC(O)-(C ₁ -C ₆ alkyl), optionally substituted -(CH ₂ ) _n C(O)-O-(C ₁ -C ₆ alkyl), optionally substituted -(CH ₂ O) _n COOH, optionally substituted -(OCH ₂ ) _n O-(C ₁ -C ₆ alkyl), optionally substituted -(CH ₂ O) _n C(O)-(C ₁ -C ₆ alkyl), optionally substituted -(OCH ₂ ) _n NHC(O)-R ₁ , optionally substituted -(CH ₂ O) _n C(O)-NR ₁ R ₂ , -(CH ₂ CH ₂ O) _n H, optionally substituted (CH ₂ CH ₂ O) _n COOH, optionally substituted -(OCH ₂ CH ₂ ) _n O-(C ₁ -C ₆ alkyl), optionally substituted -(CH ₂ CH ₂ O) _n C(O)-(C ₁ -C ₆ alkyl), optionally substituted -( OCH ₂ CH ₂ ) _n NHC(O)-R ₁ , optionally substituted (CH ₂ CH ₂ O) _n C(O)-NR ₁ R ₂ , optionally substituted -SO ₂ R _S , optionally substituted S(O)R _S , NO ₂ , CN or halogen (F, Cl, Br, I, preferably F or Cl);

R ₁ and R ₂ in ULM-g each independently represent H or a C ₁ -C ₆ alkyl group, which may be optionally substituted by one or two hydroxyl groups or up to three halogen groups (preferably fluorine);

_RS in ULM-g represents a C ₁ -C ₆ alkyl group, an optionally substituted aryl, heteroaryl, or heterocyclic group, or a -(CH ₂ ) _m NR ₁ R ₂ group;

X and X' in ULM-g are each independently C=O, C=S, -S(O), S(O) ₂ (preferably X and X' are both C=O);

R ² ' in ULM-g represents an optionally substituted -(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w alkyl group, optionally substituted -(CH ₂ ) _n -(C=O ) _u (NR ₁ ) _v (SO ₂ ) _w NR _1N R _2N group, optionally substituted -(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -aryl, optionally substituted -( CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -heteroaryl, optionally substituted -(CH ₂ ) _n -(C=O) _v NR ₁ (SO ₂ ) _w -heterocycle, optional substituted -NR ₁ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -alkyl, optionally substituted -NR ₁ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w - NR _1N R _2N , optionally substituted -NR ₁ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -NR ₁ C(O)R _1N , optional substituted -NR ₁ -(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -aryl, optionally substituted -NR ₁ -(CH ₂ ) _n -aryl-heteroaryl, optionally substituted -NR ₁ -(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -heteroaryl or optionally substituted -NR ₁ -(CH ₂ ) _n -(C=O) _v NR ₁ (SO ₂ ) _w -heterocycle, optionally substituted -X ^R2' -alkyl group; an optionally substituted -X ^R2' - aryl group; an optionally substituted -X ^R2' - heteroaryl group; an optionally substituted -X ^R2' -heterocyclic group; optionally substituted;

R ^3' in ULM-g is optionally substituted alkyl, optionally substituted -(CH ₂ ) _n -(O) _u (NR ₁ ) _v (SO ₂ ) _w -alkyl, optionally substituted -(CH ₂ ) _n -C( O) _u (NR ₁ ) _v (SO ₂ ) _w -NR _1N R _2N , optionally substituted -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -NR ₁ C(O) R _1N , optionally substituted -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -C(O)NR ₁ R ₂ , optionally substituted -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -aryl, optionally substituted -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -heteroaryl, optionally substituted -(CH ₂ ) _n - C(O) _u (NR ₁ ) _v (SO ₂ ) _w -heterocycle, optionally substituted -NR ₁ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -alkyl, optionally substituted -NR ₁ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w - NR _1N R _2N , optionally substituted -NR ₁ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -NR ₁ C(O)R _1N , optionally substituted -NR ₁ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -aryl, optionally substituted -NR ₁ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -heteroaryl, optionally substituted -NR ₁ -(CH ₂ ) _n -C(O) _u (NR ₁ ) _v (SO ₂ ) _w -heterocycle, optionally substituted -O-(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -alkyl, optionally substituted -O-(CH ₂ ) _n -( C=O) _u (NR ₁ ) _v (SO ₂ ) _w -NR _1N R _2N , optionally substituted -O-(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -NR ₁ C(O)R _1N , optionally substituted -O-(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -aryl, optionally substituted -O-(CH ₂ ) _n -( C=O) _u (NR ₁ ) _v (SO ₂ ) _w -heteroaryl or optionally substituted -O-(CH ₂ ) _n -(C=O) _u (NR ₁ ) _v (SO ₂ ) _w -heterocycle; -(CH ₂ ) _n -(V) _n' -(CH ₂ ) _n -(V) _n' -alkyl group, optionally substituted -(CH ₂ ) _n -(V) _n' -(CH ₂ ) _n -( V) _n' -aryl group, optionally substituted -(CH ₂ ) _n -(V) _n' -(CH ₂ ) _n -(V) _n' -heteroaryl group, optionally substituted -(CH ₂ ) _n -(V) _n' -(CH ₂ ) _n -(V) _n' -heterocyclic group, optionally substituted -(CH ₂ ) _n -N(R _1' )(C=O) _m' -(V) _n' -alkyl group, optionally substituted -(CH ₂ ) _n -N(R _1' )(C=O) _m' -(V) _n' -aryl group, optionally substituted -(CH ₂ ) _n -N(R _1' )(C= O) _m' -(V) _n' -heteroaryl group, optionally substituted -(CH ₂ ) _n -N(R _1' )(C=O) _m' -(V) _n' -heterocyclic group, optionally substituted -X ^R3' - alkyl group; an optionally substituted -X ^R3' -aryl group; an optionally substituted -X ^R3' - heteroaryl group; optionally substituted -X ^R3 ' - heterocyclic group; optionally substituted;

R _1N and R _2N in ULM-g are each independently H, C ₁ -C ₆ alkyl, which is optionally substituted with one or two hydroxyl groups and up to three halogen groups, or optionally substituted -(CH ₂ ) _n -aryl, -( CH ₂ ) _n -heteroaryl or -(CH ₂ ) _n -heterocyclic group;

V in ULM-g is O, S or NR ₁ ;

R ₁ in ULM-g is the same as above;

R ¹ and R _1' in ULM-g each independently represent an H or a C ₁ -C ₃ alkyl group;

X ^R2' and X ^R3' in ULM-g are each independently optionally substituted -(CH ₂ ) _n -, -(CH ₂ ) _n -CH(X _v )=CH(X _v )- (cis or trans), -(CH ₂ ) _n -CH=CH-, -(CH ₂ CH ₂ O) _n - or C ₃ -C ₆ cycloalkyl group, where X _v represents H, halogen or C ₁ -C ₃ alkyl group, which is optionally substituted;

each m in ULM-g independently represents 0, 1, 2, 3, 4, 5, 6;

each m' in ULM-g is independently 0 or 1;

each n in ULM-g independently represents 0, 1, 2, 3, 4, 5, 6;

each n' in ULM-g is independently 0 or 1;

each u in ULM-g is independently a 0 or a 1;

each v in ULM-g is independently a 0 or a 1;

each w in ULM-g is independently a 0 or a 1; And

any one or more of R ^1' , R ^2' , R ^3' , X and X' in ULM-g is optionally modified to be covalently linked to a PTM group via a linker group when PTM is not ULM' or when PTM is ULM', any one or more of R ^1' , R ^2' , R ^3' , X and X' each of ULM and ULM' are optionally modified to be covalently linked to each other directly or through a linker group, or a pharmaceutically acceptable formulation thereof salt, stereoisomer, solvate or polymorph.

[001550] In any aspect or embodiment described herein, the ULM is a cereblon E3 ligase (CLM) binding moiety selected from the group consisting of thalidomide, lenalidomide, pomalidomide, analogs thereof, isosteres thereof, or derivatives thereof.

[001551] In any aspect or embodiment described herein, CLM has a chemical structure represented by:

Where:

W is selected from the group consisting of CH ₂ , CHR, C-O, SO ₂ , NH and N-alkyl;

each X is independently selected from the group consisting of O, S and H ₂ ;

Y is selected from the group consisting of CH ₂ , -C=CR', NH, N-alkyl, N-aryl, N-hetaryl, N-cycloalkyl, N-heterocyclyl, O and S;

Z is selected from the group consisting of O, S and H ₂ ;

G and G' are independently selected from the group consisting of H, optionally substituted linear or branched alkyl, OH, R'OCOOR, R'OCONRR'', CH ₂ -heterocyclyl, optionally substituted with R', and benzyl, optionally substituted with R';

Q ₁ , Q ₂ , Q ₃ and Q ₄ represent a C carbon substituted by a group independently selected from R', N or N-oxide;

A is independently selected from the group H, optionally substituted linear or branched alkyl, cycloalkyl, C1 and F;

R contains -CONR'R'', -OR', -NR'R'', -SR', -SO ₂ R', -SO ₂ NR'R'', -CR'R''-, -CR'NR'R''-,(-CR'O)_n'R'', -aryl, -hetaryl, optionally substituted linear or branched alkyl, -cycloalkyl, -heterocyclyl, -P(O)(OR')R'' , -P(O)R'R'', -OP(O)(OR')R'', -OP(O)R'R'', -Cl, -F, -Br, -I, -CF ₃ , -CN, -NR'SO ₂ NR'R'', -NR'CONR'R'', -CONR'COR'', -NR'C(=N-CN)NR'R'', -C (=N-CN)NR'R'', -NR'C(=N-CN)R'', -NR'C(=C-NO ₂ )NR'R'', -SO ₂ NR'COR'', -NO ₂ , -CO ₂ R', -C(C=N-OR')R'', -CR'=CR'R'', -CCR', -S(C=O)(C= N-R')R'', -SF ₅ and -OCF ₃ ;

R' and R'' are independently selected from the group consisting of a bond, H, alkyl, cycloalkyl, aryl, heteroaryl, heterocycle, -C(=O)R, heterocyclyl, each of which is optionally substituted;

represents a bond that may be stereospecific ((R) or (S)) or non-stereospecific; And

Rn contains 1-4 functional groups or atoms, for example, O, OH, N, C1-C6 alkyl, C1-C6 alkoxy, -alkyl-aryl (for example, -alkyl-aryl containing at least one of C1-C6 alkyl , C4-C7 aryl or combinations thereof), aryl (eg C5-C7 aryl), amine, amide or carboxy; And

n is an integer from 1-10,

Where

when n is 1, R _n is modified to be covalently linked to a linker group (L), and

when n is 2, 3 or 4, then one R _n is modified to be covalently linked to a linker group (L) and any other R _n is optionally modified to be covalently linked to a PTM, CLM, a second CLM having the same chemical the same structure as CLM, CLM', a second linker, or any more than one or combination thereof.

[001552] In any aspect or embodiment described herein, CLM has a chemical structure represented by:

Where:

W is independently selected from CH ₂ , CHR, C=O, SO ₂ , NH and N-alkyl;

Q ₁ , Q ₂ , Q ₃ , Q ₄ , Q ₅ are each independently carbon C or N,

substituted by a group independently selected from R', N or N-oxide;

R ¹ is selected from absence, H, OH, CN, C1-C3 alkyl, C=O;

R ² is selected from the group of absence, H, OH, CN, C1-C3 alkyl, CHF ₂ , CF ₃ , CHO, C(=O)NH ₂ ;

R ³ is selected from H, alkyl (for example, C1-C6 or C1-C3 alkyl), substituted alkyl (for example, substituted C1-C6 or C1-C3 alkyl), alkoxy (for example, C1-C6 or C1-C3 alkoxy), substituted alkoxy (eg substituted C1-C6 or C1-C3 alkoxy);

R ⁴ is selected from H, alkyl, substituted alkyl;

R ⁵ and R ⁶ are each independently H, halogen, C(=O)R', CN, OH, CF ₃ ;

X represents C, CH, C=O or N;

X ₁ represents C=O, N, CH or CH ₂ ;

R' is selected from H, halogen, amine, alkyl (eg C1-C3 alkyl), substituted alkyl (eg substituted C1-C3 alkyl), alkoxy (eg C1-C3 alkoxyl), substituted alkoxy (eg substituted C1- C3 alkoxyl), NR ² R ³ , C(=O)OR ² , optionally substituted phenyl;

n represents 0-4;

represents a single or double bond; And

CLM is covalently linked to PTM, a chemical linker group (L), ULM, CLM, CLM', or combinations thereof.

[001553] In any aspect or embodiment described herein, the ULM is a (MDM2) linking moiety (MLM) with a chemical moiety selected from the group consisting of substituted imidazolines, substituted spiroindolinones, substituted pyrrolidines, substituted piperidinones, substituted morpholinones, substituted pyrrolopyrimidines, substituted imidazolopyridines, substituted thiazoloimidazoline, substituted pyrrolopyrrolidinones and substituted isoquinolinones.

[001554] In any aspect or embodiment described herein, the MLM is selected from:

Where:

R1' and R2' of formulas A-1-1 to A-1-4 (i.e., A-1-1, A-1-2, A-1-3 and A-1-4) are independently selected from a group consisting of F, Cl, Br, I, acetylene, CN, CF ₃ and NO ₂ ;

R3' is selected from the group consisting of -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH ₂ F, -OCH ₂ CH ₂ OCH ₃ and -OCH(CH ₃ ) ₂ ;

R4' of formulas A-1-1 - A-1-4 is selected from the group consisting of H, halogen, -CH ₃ , -CF ₃ , -OCH ₃ , -C(CH ₃ ) ₃ , -CH(CH ₃ ) ₂ , -cyclopropyl, -CN, -C(CH ₃ ) ₂ OH, -C(CH ₃ ) ₂ OCH ₂ CH ₃ , -C(CH ₃ ) ₂ CH ₂ OH, -C(CH ₃ ) ₂ CH ₂ OCH ₂ CH ₃ , C(CH ₃ ) ₂ CH ₂ OCH ₂ CH ₂ OH, -C(CH ₃ ) ₂ CH ₂ OCH ₂ CH ₃ , -C(CH ₃ ) ₂ CN, C(CH ₃ ) ₂ C(O )CH ₃ , -C(CH ₃ ) ₂ C(O)NHCH ₃ , -C(CH ₃ ) ₂ C(O)N(CH ₃ ) ₂ , -SCH ₃ , -SCH ₂ CH ₃ , -S(O ) ₂ CH ₃ , -S(O ₂ )CH ₂ CH ₃ , -NHC(CH ₃ ) ₃ , -N(CH ₃ ) ₂ , pyrrolidinyl and 4-morpholinyl;

R5' of formulas A-1-1 - A-1-4 is selected from the group consisting of halogen, -cyclopropyl, -S(O) ₂ CH ₃ , -S(O) ₂ CH ₂ CH ₃ , 1-pyrrolidinyl, - NH ₂ , N(CH ₃ ) ₂ and -NHC(CH ₃ ) ₃ ; And

R6' of formulas A-1-1 - A-1-4 is selected from H,

where the linker is attached to the "*" in R ^6' or to the terminal atom in R ^4' ;

R7' of formula A-4-1 to A-4-6 represents one or more (eg 1, 2, 3, 4) halogens;

R8' of formula A-4-1 to A-4-6 represents one or more groups (for example, 1, 2, 3 or 4 groups) selected from the group consisting of H, -F, -Cl, -Br, -I, -CN, -NO ₂ , ethylnyl, cyclopropyl, methyl, ethyl, isopropyl, vinyl, methoxy, ethoxy, isopropoxy, -OH, other C1-6 alkyl, other C1-6 alkenyl and C1-6 alkynyl, mono- , di- or trisubstituted;

R9' of formula A-4-1 to A-4-6 is selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, alkenyl and substituted cycloalkenyl;

Z of formula A-4-1 - A-4-6 is selected from the group consisting of H, -OSH ₃ , -OSH ₂ CH ₃ and halogen;

R10' and R11' of formula A-4-1 to A-4-6 are each independently selected from the group consisting of H, (CH ₂ ) _n -R', (CH ₂ ) _n -NR'R'', (CH ₂ ) _n -NR'COR', (CH ₂ ) _n -NR'SO ₂ R'', (CH ₂ ) _n -COOH, (CH ₂ ) _n -COOR, (CH) _n -CONR'R'', (CH ₂ ) _n -OR, (CH ₂ ) _n -SR, (CH ₂ ) _n -SOR, (CH ₂ ) _n -CH(OH)-R, (CH ₂ ) _n -COR', (CH ₂ ) _n -SO ₂ R', (CH ₂ ) _n -SONR'R', (CH ₂ ) _n -SO ₂ NR'R'', (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -R, ( CH ₂ CH ₂ O) _m -(CH ₂ ) _n -OH, (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -OR, (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -NR'R'', (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -NR'COR'', (CH ₂ CH ₂ O) _m (CH ₂ ) _n -NR'SO ₂ R', (CH ₂ CH ₂ O) _m (CH ₂ ) _n -COOH, (CH ₂ CH ₂ O) _m (CH ₂ ) _n -COOR, (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -CONR'R'', ( CH ₂ CH ₂ O) _m -(CH ₂ ) _n -SO ₂ R, (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -COR, (CH ₂ CH ₂ O) _m -(CH ₂ ) _n - SONR'R'', (CH ₂ CH ₂ O) _m -(CH ₂ ) _n -SO ₂ NR'R', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n R, (CH2) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -OH, (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -OR, (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -NR'R'', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -NRCOR'', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -NR'SO ₂ R'', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -COOH, ( CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -COOR', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -CONR'R'', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -SO ₂ R', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -COR', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -SONR'R'', (CH ₂ ) _p -(CH ₂ CH ₂ O) _m -(CH ₂ ) _n -SO ₂ NR'R'', aryl-(CH ₂ ) _n -COOH and heteroaryl-alkyl-CO-alkyl-NR'R'm, where alkyl may be substituted with OR' and heteroaryl-(CH ₂ ) _n -heterocycle, where heterocycle may be optionally substituted with alkyl, hydroxyl, COOR' and COR'; where R' and R'' are selected from H, alkyl, halogen-substituted alkyl, hydroxyl, NH2, NH(alkyl), N(alkyl) ₂ , oxo, carboxy, cycloalkyl and heteroaryl;

m, n and p independently represent 0-6;

R12' of formula A-4-1 to A-4-6 is selected from the group consisting of -O-(alkyl), -O-(alkyl)-alkoxy, -C(O)-(alkyl), -C(OH )-alkyl-alkoxy, -C(O)-NH-(alkyl), -C(O)-N-(alkyl) ₂ , -S(O)-(alkyl), S(O) ₂ -(alkyl) , -C(O)-(cyclic amine) and -O-aryl-(alkyl), -O-aryl-(alkoxy);

R1'' of formula A-4-1 to A-4-6 is selected from the group consisting of H, alkyl, aryl-substituted alkyl, alkoxy-substituted alkyl, cycloalkyl, aryl-substituted cycloalkyl and alkoxy-substituted cycloalkyl; And

denotes attachment to L.

[001555] In any aspect or embodiment described herein, the MLM is selected from:

[001556] In any aspect or embodiment described herein, the ULM is an E3 ubiquitin ligase (ILM) binding IAP fragment containing the amino acids alanine (A), valine (V), proline (P), and isoleucine (I) or their non-natural mimetics.

[001557] In any aspect or embodiment described herein, the ULM is an E3 ubiquitin ligase binding IAP fragment (ILM) containing an AVPI tetrapeptide fragment or a derivative thereof.

[001558] In any aspect or embodiment described herein, the ILM is selected from the group consisting of the chemical structures represented by formulas (I), (II), (III), (IV) and (V):

Where:

R ¹ for formulas (I), (II), (III), (IV) n (V) is selected from H or alkyl;

R ² for formulas (I), (II), (III), (IV) and (V) is selected from H or alkyl;

R ³ for formulas (I), (II), (III), (IV) and (V) is selected from H, alkyl, cycloalkyl and heterocycloalkyl;

R ⁵ and R ⁶ for formulas (I), (II), (III), (IV) and (V) are independently selected from H, alkyl, cycloalkyl, heterocycloalkyl, or more preferably, R ⁵ and R ⁶ are taken together for formulas ( I), (II), (III), (IV) and (V) to form a pyrrolidine or piperidine ring optionally further fused with 1-2 cycloalkyl, heterocycloalkyl, aryl or heteroaryl rings, each of which can then be further fused with the other a cycloalkyl, heterocycloalkyl, aryl or heteroaryl ring;

R ³ and R ⁵ for formulas (I), (II), (III), (IV) and (V) taken together may form a 5-8 membered ring, optionally further fused with 1-2 cycloalkyl, heterocycloalkyl, aryl or heteroaryl;

R ⁷ for formulas (I), (II), (III), (IV) and (V) is selected from cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, aryl-C(O)-R ⁴ , arylalkyl, heteroaryl, heteroaryl -C(O)-R ⁴ , heteroaryl-R ⁴ , heteroaryl-naphthalene, heteroarylalkyl or -C(O)NH-R ⁴ , each of which is further optionally substituted with 1-3 substituents selected from halogen, alkyl, haloalkyl, hydroxyl , alkoxy, cyano, (hetero)cycloalkyl, aryl, (hetero)aryl, -C(O)NH-R ⁴ or C(O)-R ⁴ ; And

R ⁴ is selected from alkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, aryl or bicyclic aryl, arylalkyl, heteroaryl (eg, bicyclic heteroaryl), heteroarylalkyl, further optionally substituted with 1-3 substituents as described above.

[001559] In any aspect or embodiment described herein, the ILM is selected from the group consisting of:

[001560]

[001561] In any aspect or embodiment described herein, the linker (L) contains a chemical structure element represented by the formula:

-(A ^L ) _q -,

Where:

-(A ^L ) _q - represents a group that is associated with at least one of the ULM, RTM, or both;

q is an integer greater than or equal to 1;

each A ^L is independently selected from the group consisting of a link, CR ^L1 R ^L2 , O, S, SO, SO ₂ , NR ^L3 , SO ₂ NR ^L3 , SONR ^L3 , CONR ^L3 , NR ^L3 CONR ^L4 , NR ^L3 SO ₂ NR ^L4 , CO, CR ^L1 =CR ^L2 , C≡C, SiR ^L1 R ^L2 , P(O)R ^L1 , P(O)OR ^L1 , NR ^L3 C(=NCN)NR ^L4 , NR ^L3 C(=NCN) , NR ^L3 C(=CNO ₂ )NR ^L4 , C _3-11 cycloalkyl, optionally substituted with 0-6 R ^L1 and/or R ^L2 groups, C _3-11 heterocyclyl, optionally substituted with 0-6 R ^L1 and/or R ^L2 groups, aryl, optionally substituted with 0-6 R ^L1 and/or R ^L2 groups, heteroaryl, optionally substituted with 0-6 R ^L1 and/or R ^L2 groups, wherein R ^L1 or R ^L2 are each independently optionally linked to other groups to form cycloalkyl and/or heterocyclyl fragment, optionally substituted with 0-4 R ^L5 groups; And

R ^L1 , R ^L2 , R ^L3 , R ^L4 and R ^L5 are each independently H, halogen, C _1-8 alkyl, OC _1-8 alkyl, SC _1-8 alkyl, NHC _1-8 alkyl, N(C _{1 -8} alkyl) ₂ , C _3-11 cycloalkyl, aryl, heteroaryl, C _3-11 heterocyclyl, OS _1-8 cycloalkyl, SC _1-8 cycloalkyl, NHC _1-8 cycloalkyl, N(C _1-8 cycloalkyl) ₂ , N(C _1-8 cycloalkyl)(C _1-8 alkyl), OH, NH ₂ , SH, SO ₂ C _1-8 alkyl, P(O)(OS _1-8 alkyl)(C _1-8 alkyl), P(O)(OS _1-8 alkyl) ₂ , CC-C _1-8 alkyl, CCH, CH=CH(C _1-8 alkyl), C(C _1-8 alkyl)=CH(C _1-8 alkyl ), C(C _1-8 alkyl)=C(C _1-8 alkyl) ₂ , Si(OH) ₃ , Si(C _1-8 alkyl) ₃ , Si(OH)(C _1-8 alkyl) ₂ , SOS _1-8 alkyl, CO ₂ H, halogen, CN, CF ₃ , CHF ₂ , CH ₂ F, NO ₂ , SF ₅ , SO ₂ NHC _1-8 alkyl, SO ₂ N(C _1-8 alkyl) ₂ , SONHC _1-8 alkyl, SON(C _1-8 alkyl) ₂ , CONHC _1-8 alkyl, CON(C _1-8 alkyl) ₂ , N(C _1-8 alkyl)CONH(C _1-8 alkyl), N (C _1-8 alkyl)CON(C _1-8 alkyl) ₂ , NHCONH(C _1-8 alkyl), NHCON(C _1-8 alkyl) ₂ , NHCONH ₂ , N(C _1-8 alkyl)SO ₂ NH (C _1-8 alkyl), N(C _1-8 alkyl) SO ₂ N(C _1-8 alkyl) ₂ , NH SO ₂ NH(C _1-8 alkyl), NH SO ₂ N(C _1-8 alkyl ) ₂ , NH SO ₂ NH ₂ .

[001562] In any aspect or embodiment described herein, the linker (L) includes an optionally substituted C ₁ -C ₅₀ alkyl (for example, C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , C ₇ , C ₈ , C ₉ , C ₁₀ , C ₁₁ , C ₁₂ , C ₁₃ , C ₁₄ , C ₁₅ , C 16, C ₁₇ , C ₁₈ , C ₁₉ , C ₂₀ , _{C 21 ,} C ₂₂ , C ₂₃ , С ₂₄ , С ₂₅ , С ₂₆ , С ₂₇ , С ₂₈ , С ₂₉ , С ₃₀ , С _{31, С 32, С 33 ,} С ₃₄ , С ₃₅ , С ₃₆ , С ₃₇ , С ₃₈ , _{С 39 ,} С ₄₀ , C ₄₁ , C ₄₂ , C ₄₃ , C ₄₄ , C ₄₅ , C ₄₆ , C ₄₇ , C ₄₈ , C ₄₉ or C ₅₀ alkyl), where each carbon is optionally replaced by (1) a heteroatom selected from N, S, P or Si, which have the appropriate number of hydrogen atoms, substitutions, or both for total valency, (2) optionally substituted cycloalkyl or bicyclic cycloalkyl, (3) optionally substituted heterocycloalkyl or bicyclic heterocycloalkyl, (4) optionally substituted aryl bicyclic aryl, or ( 5) optionally substituted heteroaryl or bicyclic heteroaryl, provided that there is no heteroatom-heteroatom bond (for example, the heteroatoms are not covalently bonded or located adjacent to each other).

[001563] In any aspect or embodiment described herein, the linker element A ^L (L) contains a group represented by a general structure selected from the group consisting of:

-N(R)-(CH2) _m -O(CH2) _n -O(CH2) _o -O(CH2) _p -O(CH2) _q -O(CH2) _r -OCH2-,

-O-(CH2) _m -O(CH2) _n -O(CH2) _o -O(CH2) _p -O(CH2) _q -O(CH2) _r -OCH2-,

-O-(CH2) _m -O(CH2) _n -O(CH2) _o -O(CH2) _p -O(CH2) _q -O(CH2) _r -O-;

-N(R)-(CH2) _m -O(CH2) _n -O(CH2) _o -O(CH2) _p -O(CH2) _q -O(CH2)rO-;

-(CH2) _m -O(CH ₂ ) _n -O(CH2) _o -O(CH2) _p -O(CH2) _q -O(CH2) _r -O-;

-(CH2) _m -O(CH ₂ ) _n -O(CH2) _o -O(CH2) _p -O(CH2) _q -O(CH2) _r -OCH2-;

where each m, n, o, p, q and r, independently represents 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 , 17, 18, 19 or 20, with the proviso that when the amount is zero, there is no N-O or O-O bond, R is selected from the group of H, methyl and ethyl, and X is selected from the group of H and F;

[001564] In any aspect or embodiment described herein, the linker element A ^L (L) is selected from the group consisting of:

where each m and n are independently selected from 0, 1, 2, 3, 4, 5 or 6.

[001565] In any aspect or embodiment described herein, the linker element A ^L (L) is selected from the group consisting of:

where each m, n, o, p, q, r and s independently represents 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.

[001566] In any aspect or embodiment described herein, the linker element A ^L is selected from:

[001567] In any aspect or embodiment described herein, the linker (L) is a polyethyleneoxy group, optionally substituted with aryl or phenyl, containing from 1 to 10 ethylene glycol units.

[001568] In any aspect or embodiment described herein, the linker (L) contains the following chemical structure:

Where:

W ^L1 and W ^L2 are each independently absent, a 4-8 membered ring with 0-4 heteroatoms, optionally substituted with RQ, wherein each RQ is independently H, halogen, OH, CN, CF3, optionally substituted linear or branched C1- C6 alkyl, optionally substituted linear or branched C1-C6 alkoxy, or 2 RQ groups taken together with the atom to which they are attached to form a 4-8 membered ring system containing 0-4 heteroatoms;

Y ^L1 each independently represents a bond, optionally substituted straight or branched C1-C6 alkyl, and optionally one or more C atoms are replaced by O; or optionally substituted linear or branched C1-C6 alkoxy;

n represents 0-10; And

denotes the point of attachment to PTM or ULM fragments.

[001569] In any aspect or embodiment described herein, the linker (L) contains the following chemical structure:

Where:

W ^L1 and W ^L2 are each independently absent, represent aryl, heteroaryl, cyclic group, heterocycle, C _1-6 alkyl and optionally one or more C atoms replaced by O or N, C _1-6 alkene and optionally one or more C atoms replaced by O, a C _1-6 alkyne and optionally one or more C atoms are replaced by O, a bicyclic group, biaryl, biheteroaryl or biheterocycle, each optionally substituted by R ^Q , each R ^Q independently represents H, halogen, OH, CN, CF ₃ , hydroxyl, nitrogen, C≡CH, C _2-6 alkenyl, C _2-6 alkynyl, optionally substituted linear or branched C ₁ -C ₆ alkyl, optionally substituted linear or branched C ₁ -C ₆ alkoxy, optionally substituted OS _1-3 alkyl (e.g., optionally substituted with 1 or more -F), OH, NH ₂ , NR ^Y1 R ^Y2 , CN or 2 R ^Q groups taken together with the atom to which they are attached to form a 4-8 membered ring system , containing 0-4 heteroatoms; YL1 is each independently a bond, NR ^YL1 , O, S, NR ^YL2 , CR ^YL1 R ^YL2 , C=O, C=S, SO, SO ₂ , optionally substituted linear or branched C ₁ -C ₆ alkyl, and optionally one or more C atoms are replaced by O; optionally substituted linear or branched C ₁ -C ₆ alkoxy, 3-6 membered acyclic or aromatic ring with 0-4 heteroatoms;

Q ^L is a 3-6 membered alicyclic or aromatic ring with 0-4 heteroatoms, optionally bridged, optionally substituted with 0-6 R ^Q , each R ^Q independently represents H, optionally substituted linear or branched C _1-6 alkyl (eg , optionally substituted with 1 or more halogens or C _1-6 alkoxyl), or 2 R ^Q groups taken together with the atom to which they are attached to form a 3-8 membered ring system containing 0-2 heteroatoms;

R ^YL1 , R ^YL2 each independently represents H, OH, optionally substituted linear or branched C _1-6 alkyl (for example, optionally substituted with 1 or more halogens or C _1-6 alkoxy), or R ¹ , R ² together with an atom, to which they join, form a system of 3-8 membered rings containing 0-2 heteroatoms;

n represents 0-10; And

denotes the point of attachment to PTM or ULM fragments.

[001570] In any aspect or embodiment described herein, the linker element A ^L (L) is selected from the group consisting of:

[001571] In any aspect or embodiment described herein, at least one of:

(a) the PTM is selected from the combination of tables 4, 6, 8, 10 and 12 or the PTM of table 1;

(b) the ULM is selected from the combination of tables 4, 6, 8, 10 and 12 or the ULM of table 3;

(c) the linker element A ^L (L) is selected from:

(d) combinations thereof, where:

*N of heterocycloalkyl shared with PTM; A

each m, n, o, p, q, r and s is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[001572] In any aspect or embodiment described herein, the linker element A ^L or L is selected from the group consisting of:

where N* of the heterocycloalkyl is shared with PTM.

[001573] In any aspect or embodiment described herein, at least one of:

The PTM is selected from PTM-1, PTM-2, PTM-3, PTM-4, PTM-5 or PTM-6;

the linker is selected from L-1, L-2, L-3, L-4, L-5 or L-6;

The ULM is selected from ULM-1, ULM-2, ULM-3, ULM-4 or ULM-5; or

their combinations.

[001574] In any aspect or embodiment described herein, the compound is selected from the group consisting of: exemplary compounds 1-10.

[001575] In another aspect, the present invention provides a composition comprising an effective amount of a bifunctional compound of the present invention and a pharmaceutically acceptable carrier.

[001576] In any aspect or embodiment described herein, the composition further comprises at least one additional bioactive agent or other bifunctional compound of the present invention.

[001577] In any aspect or embodiment described herein, the additional bioactive agent is an anticancer agent (eg, an epidermal growth factor receptor inhibitor).

[001578] A further aspect of the present invention provides a composition comprising a pharmaceutically acceptable carrier and an effective amount of at least one compound of the present invention for treating a disease or disorder in a subject, the method comprising administering the composition to a subject in need thereof, wherein the compound is effective for treating or alleviation of at least one symptom of the disease or disorder.

[001579] In any aspect or embodiment described herein, the disease or disorder is associated with at least one of KRas accumulation, aggregation, overactivation, or combinations thereof.

[001580] In any aspect or embodiment described herein, the disease or disorder is a cancer that is associated with the accumulation, aggregation and/or overactivation of KRas.

[001581] In any aspect or embodiment described herein, the disease or disorder is pancreatic cancer, colon cancer, colorectal cancer, lung cancer, non-small cell lung cancer, biliary tract malignancies, endometrial cancer, cervical cancer, bladder cancer, liver cancer, myeloid leukemia and breast cancer.

[001582] In any aspect or embodiment described herein, the disease or disorder is pancreatic cancer, colon cancer, lung cancer, non-small cell lung cancer.

EXAMPLES

Analyzes and degradation data

[001583] Cell lines, antibodies and reagents. NCI-H2030 (CRL-5914) and SW1573 (CRL-2170) were purchased from ATCC. Both cell lines are homozygous for the G12C mutation in KRas. NCI-H2030 cells were cultured in RPMI-1640 medium containing 1% penicillin-streptomycin and 10% fetal bovine serum (FBS). SW1573 cells were cultured in DMEM containing 1% penicillin-streptomycin and 10% FBS. Antibodies for KRas detection (cat. no. LS-C175665) were purchased from LifeSpan Biosciences and used at a dilution of 1:2000. GAPDH was detected using an antibody purchased from Cell Signaling Technology (cat. no. 5174) at a dilution of 1:3000. Secondary anti-mouse and anti-rabbit detection antibodies were purchased from Cell Signaling Technology (cat. nos. 7076 and 7074, respectively).

[001584] Compound treatment and Western blotting according to Table 5. H2030 cells in 12-well plates were serum starved for 24 hours and then treated with 0.3 μM, 1 μM, and 3 μM of the indicated bifunctional compound for 24 hours. Cells were lysed in Cell Signaling Technology Cell Lysis Buffer (Cat #9803) supplemented with protease inhibitors, and proteins were separated by SDS-PAGE. KRas was detected by immunoblotting using the LSBio antibody (cat. no. LS-C175665).

[001585] Compound treatment and Western blotting according to Table 14. NCI-H2030 or SW1573 cells were plated in 12-well plates and allowed to adhere overnight at 37°C in an incubator containing 5% CO2. The next day, the medium was replaced with the corresponding medium without FBS to induce starvation. Cells were returned to the incubator for another 24 hours after changing the medium. Compounds were then added to the appropriate final concentration (0.3 μM, 1 μM, and 3 μM) in 0.1% DMSO. Cells were treated with VHL-based compounds for 24 hours and all other compounds for 72 hours. After treatment, cells were lysed in RIPA buffer (Thermo Fisher cat. no. 89900) containing phosphatase (Thermo Fisher cat. no. 1861277) and protease inhibitor cocktails (Thermo Fisher cat. no. 78429). Lysates were cleared at 13,000 rpm for 15 minutes, and supernatants were analyzed for total protein concentration using a Pierce BCA assay system (Thermo Fisher cat. #23225). For each sample, 10 μg of total protein was separated on a 12% Bis-Tris gel and then transferred to a nitrocellulose membrane. After blocking in 3% BSA in TBST (Tris-buffered saline with 0.1% Tween 20) for 1 hour at room temperature, membranes were probed with primary antibody (LS Bio cat. no. LS-C175665) overnight at 4°C. C. After incubation with the secondary antibody, the membranes were visualized the next day using SuperSignal™ West Femto Substrate (Thermo Fisher cat. #34095). The KRas signal in each lane was normalized to GAPDH, and the percentage of degradation was estimated to be less than 25% by visual inspection (C in Table 13), or quantified relative to the DMSO control lane using BIO-RAD Image Lab 5.2.1, and all data were built using GraphPad PRISM 6.07.

[001586] DC ₅₀ represents the half-maximal degradation concentration, that is, the concentration at which 50% degradation occurs. D _Max represents the maximum achievable degradation efficiency (i.e., maximum observed degradation) in the degradation data.

[001587] Figures 2A and 2B show the result of treating cells with either the bifunctional compound alone or the bifunctional compound and an E3 ubiquitin ligase inhibitor and then detecting KRas protein on the gel. The mobility shift in the detection of KRas on the gel shows that the bifunctional compounds covalently modify KRas. The top band represents KRas and the bifunctional compound, and the bottom band represents only KRas. Figure 2A is a representative Western blot of a highly potent disruptor, representative compound 399. Figure 2B is a representative Western blot of a less potent disruptor, representative compound 432. Both compounds covalently modify KRas ^G12C , as indicated by gel shift.

[001588] The bifunctional compounds of Tables 4, 6, 8, 10 and 12 demonstrated degradation of the target protein when tested under the conditions described above. For example, each of the bifunctional compounds from Table 4 had DC ₅₀ ranging from 500 nM to 1 μM and D _max as shown in Table 5. As a further example, the bifunctional compounds of Tables 6, 8, 10 and 12 resulted in KRas degradation. as shown in Table 14. In the tables, “BUT” means that a specific parameter, characteristic, etc. has not been defined for a specific compound.

[001589] A novel bifunctional molecule is described that contains a KRas-recruiting moiety and an E3 ligase (eg, CLM, VLM, ILM, or MLM) recruiting moiety through PROTAC technology. Bifunctional molecules according to the present invention actively destroy KRas, which leads to a strong suppression of cell proliferation and induction of apoptosis. PROTAC-mediated protein degradation represents a promising strategy to target “undruggable” pathological proteins such as KRas using traditional approaches.

[001590] The contents of all references, patents, pending patent applications and published patents cited in this application are hereby incorporated by reference in their entirety.

[001591] Those skilled in the art will recognize or be able to determine, using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be covered by the following claims. It is understood that the detailed examples and embodiments described herein are provided by way of example only for illustrative purposes and are not intended to limit the invention in any way. In light of this, various modifications or changes may be suggested to those skilled in the art, are included within the spirit and scope of this application, and are contemplated within the scope of the appended claims. For example, the relative amounts of ingredients may be changed to optimize desired effects, additional ingredients may be added, and/or similar ingredients may be replaced by one or more of the described ingredients. Additional useful features and functionality associated with the systems, methods and processes of the present invention will be apparent from the accompanying claims. Moreover, those skilled in the art will recognize or be able to determine, using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be covered by the following claims.

Cited links:

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Ma Y, Gu Y, Zhang Q, Han Y, Yu S, Lu Z, Chen J. Targeted degradation of KRAS by an engineered ubiquitin ligase suppresses pancreatic cancer cell growth in vitro and in vivo. Mol Cancer Ther. Mar 2013; 12(3):286-94.

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Yuan TL, Fellmann C, Lee CS, Ritchie CD, Thapar V, Lee LC, Hsu DJ, Grace D, Carver JO, Zuber J, Luo J, McCormick F, Lowe SW. Development of siRNA payloads to target KRAS-mutant cancer. Cancer Discov. 2014 Oct; 4(10):1182-1197.

Claims (133)

1. A compound having a chemical structure ULM-L-PTM, or a pharmaceutically acceptable salt thereof, where (a) L is represented by the formula -(A ^L ) _q -, where q is an integer from 1 to 50; each A ^L is independently selected from the group consisting of CR ^L1 R ^L2 , O, NR ^L3 , CO, C≡C, C _3-11 heterocycloalkyl containing 1 or 2 N atoms, optionally substituted with 1-3 R ^L1 groups and/or R ^L2 , and phenyl optionally substituted with 1-3 R ^L1 and/or R ^L2 groups; And R ^L1 , R ^L2 and R ^L3 are each independently H or C _1-8 alkyl; (b) ULM is , where W ³ is selected from , 5-10 membered heteroaryl or phenyl, wherein the heteroaryl contains 1 or 2 atoms independently selected from N, O or S, and wherein the heteroaryl and phenyl are optionally substituted with 1 or 2 C _1-6 alkyls; R ₉ and R ₁₀ independently represent hydrogen or C _1-6 alkyl; R ₁₁ represents or 5-10 membered heteroaryl, optionally substituted with 1 or 2 C _1-6 alkyls, the heteroaryl containing 1 or 2 atoms selected from N, O or S; R ₁₂ represents H or C _1-6 alkyl; R ₁₃ represents H, C _1-6 alkyl or C _1-6 alkylcarbonyl; each of R _14a, R _14b independently represents H or C _1-6 alkyl; W ⁵ represents a 5-10 membered heteroaryl or phenyl, wherein the heteroaryl contains 1 or 2 atoms independently selected from N, O or S, and wherein the heteroaryl and phenyl are optionally substituted with 1 or 2 C _1-6 alkyls; R ₁₅ represents H or 5-10 membered heteroaryl, optionally substituted with 1 or 2 C _1-6 alkyls, the heteroaryl containing 1 or 2 atoms selected from N, O or S; each R ₁₆ independently represents C _1-6 alkyl or C _1-6 alkoxy; o represents 0, 1, 2, 3 or 4; denotes the attachment point L; (c) PTM selected from Where represents phenyl or heterocycloalkyl, wherein heterocycloalkyl contains 1 N atom; X _PTM is N; R _PTM1A is NR _PTM9 R _PTM10 , O-(C _3-6 heterocycloalkyl), wherein the heterocycloalkyl contains 1 N atom, -OC _1-4 alkyl-C _3-6 heterocycloalkyl, optionally substituted with 1 or 2 substituents independently selected from C _1-6 alkyl or O-C _1-6 alkyl, wherein heterocycloalkyl contains 1 or 2 N atoms, , or wherein *carbon is replaced by two hydrogen atoms or a hydrogen atom and a C _1-6 alkyl and N* represents the N atom of a heterocycloalkyl contained in L, wherein the heterocycloalkyl contains 1 or 2 N atoms; R _PTM1B is H or ; each of R _PTM9 and R _PTM10 independently represents H, phenyl or C _1-6 alkyl, optionally substituted with 1 or 2 substituents independently selected from C(O)NH ₂ , C(O)N(C _1-6 alkyl) ₂ and C(O)N(H)(C _1-6 alkyl); R _PTM2 is H or (C=O)R _PTM2' ; R _PTM2' is C _1-6 alkyl, C _2-6 alkene, -N(R _PTM8 ) ₂ or -C(OH) ₂ ; R _PTM3 is phenyl or naphthalenyl, each independently substituted with 1 or 2 substituents independently selected from OH, H and halogen; R _PTM4A represents 1 or 2 substituents independently selected from OH, H and halogen; R _PTM4B is H or -CH ₂ CN; R _PTM8 is H or C _1-6 alkyl; t represents 0, 1, 2, 3, 4, 5, 6; And denotes the attachment site of L. 2. The compound according to claim 1, characterized in that L represents C ₁ -C ₅₀ alkyl, where each carbon is part of a methylene group or is replaced by N(H), N(C _1-6 alkyl) or phenyl. 3. Connection selected from: or a pharmaceutically acceptable salt thereof. 4. Connection according to any one of paragraphs. 1, 2, characterized in that the PTM is selected from: 5. Connection according to any one of paragraphs. 1-4, characterized in that the ULM is selected from 6. Connection according to any one of paragraphs. 1, 4 and 5, characterized in that L is selected from , 7. A pharmaceutical composition for the degradation of Kirsten rat sarcoma protein (KRas), containing an effective amount of a compound according to claim 1 or 3 and a pharmaceutically acceptable excipient. 8. Use of an effective amount of a compound according to claim 1 or 3 or a pharmaceutical composition according to claim 7 for treating in a subject a disease or disorder associated with the accumulation, aggregation and/or overactivation of Kirsten rat sarcoma protein (KRas). 9. The use of claim 8, wherein the disease or disorder associated with the accumulation, aggregation and/or overactivation of KRas is cancer. 10. Use according to claim 8 or 9, characterized in that the disease or disorder associated with the accumulation, aggregation and/or overactivation of KRas is pancreatic cancer, colon cancer, colorectal cancer, lung cancer, non-small cell lung cancer, malignant biliary tract neoplasms, endometrial cancer, cervical cancer, bladder cancer, liver cancer, myeloid leukemia and breast cancer. 11. Application according to any one of paragraphs. 8 or 9, wherein the disease or disorder associated with the accumulation, aggregation and/or overactivation of KRas is selected from pancreatic cancer, colon cancer, lung cancer and non-small cell lung cancer.