Compounds and methods for targeted degradation of KRAS

Abstract

The problem that the present invention aims to solve is to provide a heterobifunctional compound containing a target protein binding site and an E3 ubiquitin ligase binding site, as well as related methods of use. [Solution] The present invention relates to a bifunctional compound used as a regulator of Carsten Ras sarcoma protein (KRas or KRAS). In particular, the hetero-bifunctional compound of the present invention contains a site that binds to von Hippel-Lindau E3 ubiquitin ligase at one end and a site that binds to KRas at the other end, so that the target protein is positioned near the ubiquitin ligase, resulting in the degradation (and inhibition) of the target protein. The hetero-bifunctional compound of the present invention exhibits a broad range of pharmacological activity related to the degradation / inhibition of the target protein. Diseases or disorders resulting from abnormal control of the target protein can be treated or prevented with the compounds and compositions of the present invention.

Description

[Technical Field]

[0001] Cross-reference of related applications This disclosure claims the benefit and priority of U.S. Provisional Application No. 63 / 006,000, filed on April 6, 2020, entitled “COMPOUNDS AND METHODS FOR TARGETED DEGRADATION OF KRAS,” and U.S. Provisional Application No. 63 / 030440, filed on May 27, 2020, entitled “COMPOUNDS AND METHODS FOR TARGETED DEGRADATION OF KRAS” (which is incorporated herein by reference in its entirety for all purposes).

[0002] Built-in by reference All cited references, including U.S. Patent Application No. 15 / 074820, filed on 18 March 2016 and published as U.S. Patent Application Publication No. 2016 / 0272639; U.S. Patent Application No. 14 / 371956, filed on 11 July 2014 and published as U.S. Patent Application Publication No. 2014 / 0356322; U.S. Patent Application No. 16 / 224088, filed on 18 December 2018 and published as U.S. Patent Application Publication No. 2019 / 0127359; and U.S. Patent Application No. 16 / 375643, filed on 4 April 2019 and published as U.S. Patent Application Publication No. 2019 / 0315732, are incorporated herein by reference in their entirety.

[0003] Description of research funded by the federal government. This invention was made with government support under National Institutes of Health authorization number NIH R35CA197589. The government reserves certain rights in this invention.

[0004] This invention provides a heterobifunctional compound comprising a target protein binding site and an E3 ubiquitin ligase binding site, as well as related methods of use. The bifunctional compound is useful as a modulator of targeted ubiquitination of the Karsten ras sarcoma protein having a G12C mutation that is subsequently degraded and / or inhibited. [Background technology]

[0005] Most small molecule drugs bind to enzymes or receptors in tight, clearly defined pockets. Protein-protein interactions, on the other hand, are notoriously difficult to target using small molecules due to their large contact surfaces and the shallow grooves or flat interfaces involved. E3 ubiquitin ligases (hundreds of which are known in humans) confer substrate specificity for ubiquitination and are therefore more attractive therapeutic targets than common proteasome inhibitors due to their specificity for a given protein substrate. The development of ligands for E3 ligases has proven difficult, partly due to the fact that they must interfere with protein-protein interactions. However, recent developments have provided specific ligands that bind to these ligases. For example, from the discovery of nutrin, the first small molecule E3 ligase inhibitor, to the reporting of further compounds targeting E3 ligases,

[0006] The von Hippel-Lindau (VHL) tumor suppressor is a substrate-recognizing subunit of the E3 ligase complex VCB, consisting of elongin B and C, Cul2, and Rbx1. The major substrate of VHL is hypoxia-inducible factor 1α (HIF-1α), a transcription factor that upregulates genes such as the pro-angiogenic growth factor VEGF and the erythropoietin-inducing cytokine in response to hypoxic levels. The first small molecule ligand of von Hippel-Lindau (VHL) was generated for the substrate-recognizing subunit of the E3 ligase, and a crystal structure was obtained confirming that the compound mimics the binding mode of the transcription factor HIF-1α, the major substrate of VHL.

[0007] Bifunctional compounds, such as those described in U.S. Patent Application Publications 2015 / 0291562 and 2014 / 0356322 (incorporated herein by reference), function to recruit endogenous proteins to E3 ubiquitin ligases for ubiquitination and subsequent degradation in the proteasome degradation pathway. In particular, the publications cited above describe bifunctional or proteolytically targeted chimeric (PROTAC®) proteolytic compounds, which are used as modulators of targeted ubiquitination of a variety of polypeptides and proteins that are subsequently degraded and / or inhibited by bifunctional compounds.

[0008] The Carsten rat sarcoma (KRAS) gene is an oncogene that encodes KRas, a low molecular weight GTPase signaling protein. The Ras protein associates with the cell membrane and acts as a switch in the transmission of extracellular signals to intracellular responses, thereby regulating, for example, cell division. In normal cells, KRAS functions as a molecular switch, circulating between an "off" state bound to inactive GDP and an "on" state bound to active GTP (Milburn et al.; Ito, Y., et al., Regional polysterism in the GTP-bound form of the human c-Ha-Ras protein. Biochemistry 1997, 36(30), 9109-9119). This switch is tightly regulated by guanine nucleotide exchange factor (GEF) proteins that exchange GDP for GTP, and GTPase-activating proteins (GAPs) that enhance the inherently slow GTPase activity of KRAS (Bar-Sagi, D., The Sos (Son of Sevenless) protein. Trends Endocrinol Metab 1994, 5(4), 165-9; Pierre, S., et al., Understanding SOS (Son of Sevenless). Biochem Pharmacol 2011, 82(9), 1049-56; Harrell Stewart, DR, et al., Pumping the brakes on RAS - negative regulators and death effectors of RAS. J Cell Sci 2020, 133(3)).GEF and GAP effector proteins bind to one or both of two shallow binding pockets on KRAS, referred to as switch I (residues 30-38) and switch II (residues 59-76), and their conformation changes dramatically between GDP-bound and GTP-bound states (Ito et al.; Boriack-Sjodin, PA et al., The structural basis of the activation of Ras by Sos. Nature 1998, 394(6691), 337-43; Scheffzek, K. et al., The Ras-RasGAP complex: structural basis for GTPase activation and its loss in oncogenic Ras mutants. Science 1997, 277(5324), 333-8).

[0009] The KRAS gene is one of the most frequently mutated oncogenes in cancer (Prior, IA; Lewis, PD; Mattos, C., A comprehensive survey of Ras mutations in cancer. Cancer Res 2012, 72(10), 2457-67; Land, H.; Parada, LF; Weinberg, RA, Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes. Nature 1983, 304(5927), 596-602; Newbold, RF; Overell, RW, Fibroblast Immortality Is a Prerequisite for Transformation by Ej C-Ha-Ras Oncogene. Nature 1983, 304(5927), 648-651). KRAS encodes a small, membrane-bound GTPase that relays signals from receptor tyrosine kinases (RTKs), promoting cell proliferation, cell differentiation, or cell death(Milburn,MV,et al.,Molecular Switch for Signal Transduction - Structural Differences between Active and Inactive Forms of Protooncogenic Ras Proteins.Science 1990,247(4945),939-945;Simanshu,DK,et al.,RAS Proteins and Their Regulators in Human Disease.Cell 2017, 170(1), 17-33).Somatic KRAS mutations reduce the protein's GAP-mediated enzyme activity, leading to the accumulation of GTP-bound active KRAS and hyperactivation of downstream signaling, which can result in uncontrolled cell proliferation (Prior et al.; Simanshu et al.). Numerous activating or gain-of-function mutations in the KRas gene are known, and indeed, KRas is the most frequently mutated gene in cancer. Gain-of-function KRas 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 KRas' ability to switch between active and inactive states. The crucial role of mutant KRas is established in the onset, maintenance, progression, and metastasis of various cancers, and mutations frequently correlate with poor prognosis and increased resistance to chemotherapy and biological therapies, including those targeting the epidermal growth factor receptor. However, despite its critical role and high frequency in cancer, there are no effective therapies that directly target this oncogene, leading to the perception that it is "undrug-proof." Furthermore, despite its high prevalence in cancer and years of extensive research efforts, mutant KRAS remains a challenging therapeutic target, given the lack of conventional drug-prone pockets on its surface (Spencer-Smith, R. et al., Direct inhibition of RAS: Quest for the Holy Grail? Semin Cancer Biol 2019, 54, 138-148).

[0010] KRAS G12C mutations are frequently observed in lung adenocarcinoma (LUAD). G12C KRAS variants constitute over 50% of all KRAS-mutant LUAD tumors (13% of all LUAD tumors) (Prior et al. 2012). Additionally, 3% of colorectal cancers and 1% of all other solid tumors are KRAS-mutant. G12CThis mutation expresses KRAS (Campbell, JD, et al., Distinct patterns of somatic genome alterations in lung adenocarcinomas and squamous cell carcinomas. Nat Genet 2016, 48(6), 607-16). This mutation significantly reduces the intrinsic GTPase activity of KRAS, enabling the accumulation of GTP-bound KRAS (Lu, S., et al., GTP Binding and Oncogenic Mutations May Attenuate Hypervariable Region (HVR)-Catalytic Domain Interactions in Small GTPase K-Ras4B, Exposing the Effector Binding Site. J Biol Chem 2015, 290(48), 28887-900). Recent advances, first drawn by Shokat's group, are related to KRAS G12CMolecules that covalently and selectively bind to mutated cysteine ​​were identified (Ostrem, JM, et al., K-Ras(G12C) inhibitors allosterically control GTP affinity and effector interactions. Nature 2013, 503(7477), 548-51; Rudolph, J., et al., Selective inhibition of mutant Ras protein through covalent binding. Angew Chem Int Ed Engl 2014, 53(15), 3777-9; Ostrem, JM, et al., Direct small-molecule inhibitors of KRAS: from structural insights to mechanism-based design. Nat Rev Drug Discov 2016, 15(11), 771-785; Nnadi, CI, et al., Novel K-Ras G12C Switch-II Covalent Binders Destabilize Ras and Accelerate Nucleotide Exchange. J Chem Inf Model (2018, 58(2), 464-471). These compounds induce a novel drug-like pocket within the KRAS switch II region (Ostrem et al. 2013). Optimization of electrophilic interactions contributing to cysteine ​​conjugation and molecular interactions within the drug induction pocket leads to orally bioavailable KRAS G12CThis has led to the development of inhibitors. However, despite this success, rapid adaptive resistance and MAPK signaling reactivation after inhibitor treatment have already been reported (Ryan, MB et al., Vertical Pathway Inhibition Overcomes Adaptive Feedback Resistance to KRAS(G12C) Inhibition. Clin Cancer Res 2019; Xue, JY, et al., Rapid non-uniform adaptation to conformation-specific KRAS(G12C) Inhibition. Nature 2020, 577(7790), 421-425). Therefore, the development of complementary therapeutic strategies may help realize the full potential of targeting KRAS variants for cancer treatment.

[0011] There is a continuing need in the art for effective treatments for KRas-related diseases and disorders, such as 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. [Prior art documents] [Patent Documents]

[0012] [Patent Document 1] U.S. Provisional Application No. 63 / 006,000 [Patent Document 2] U.S. Provisional Application No. 63 / 030440 [Patent Document 3] U.S. Patent Application No. 15 / 074820, filed on March 18, 2016, was published as U.S. Patent Application Publication No. 2016 / 0272639. [Patent Document 4] U.S. Patent Application No. 14 / 371956, filed on July 11, 2014, was published as U.S. Patent Application Publication No. 2014 / 0356322. [Patent Document 5] U.S. Patent Application No. 16 / 224088, filed on December 18, 2018, was published as U.S. Patent Application Publication No. 2019 / 0127359. [Patent Document 6] U.S. Patent Application No. 16 / 375643, filed on April 4, 2019, was published as U.S. Patent Application Publication No. 2019 / 0315732. [Patent Document 7] U.S. Patent Application Publication 2015 / 0291562 [Patent Document 8] U.S. Patent Application Publication 2014 / 0356322 [Non-patent literature]

[0013] [Non-Patent Document 1] Milburn et al.;Ito,Y.,et al.,Regional polysterism in the GTP-bound form of the human c-Ha-Ras protein.Biochemistry 1997,36(30),9109-9119 [Non-Patent Document 2] Bar-Sagi, D., The Sos(Son of sevenless) protein.Trends Endocrinol Metab 1994,5(4),165-9 [Non-Patent Document 3] Pierre, S., et al., Understanding SOS (Son of Sevenless).Biochem Pharmacol 2011, 82(9), 1049-56 [Non-Patent Document 4] Harrell Stewart,DR,et al.,Pumping the brakes on RAS - negative regulators and death effectors of RAS.J Cell Sci 2020,133(3) [Non-Patent Document 5] Ito et al.;Boriack-Sjodin,PAet al.,The structural basis of the activation of Ras by Sos.Nature 1998,394(6691),337-43 [Non-Patent Document 6] Scheffzek, K. et al., The Ras-RasGAP complex: structural basis for GTPase activation and its loss in oncogenic Ras mutants.Science 1997, 277(5324), 333-8 [Non-Patent Document 7] Prior,IA;Lewis,PD;Mattos,C.,A comprehensive survey of Ras mutations in cancer.Cancer Res 2012,72(10),2457-67 [Non-Patent Document 8] Land, H.;Parada,LF;Weinberg,RA,Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes.Nature 1983,304(5927),596-602 [Non-Patent Document 9] Newbold,RF;Overell,RW,Fibroblast Immortality Is a Prerequisite for Transformation by Ej C-Ha-Ras Oncogene.Nature 1983,304(5927),648-651 [Non-Patent Document 10] KRAS encodes a small, membrane bound GTPase that relays signals from receptor tyrosine kinases (RTKs), promoting cell proliferation, cell differentiation or cell death (Milburn, M.V., et al., Molecular Switch for Signal Transduction - Structural Differences between Active and Inactive Forms of Protooncogenic Ras Proteins. Science 1990, 247(4945), 939-945 [Non-Patent Document 11] Simanshu, D.K., et al., RAS Proteins and Their Regulators in Human Disease. Cell 2017, 170(1), 17-33 [Non-Patent Document 12] Spencer-Smith, R. et al., Direct inhibition of RAS: Quest for the Holy Grail? Semin Cancer Biol 2019, 54, 138-148 [Non-Patent Document 13] Campbell, J.D., et al.., Distinct patterns of somatic genome alterations in lung adenocarcinomas and squamous cell carcinomas. Nat Genet 2016, 48(6), 607-16 [Non-Patent Document 14] Lu,S.,et al.,GTP Binding and Oncogenic Mutations May Attenuate Hypervariable Region(HVR)-Catalytic Domain Interactions in Small GTPase K-Ras4B,Exposing the Effector Binding Site.J Biol Chem 2015,290(48),28887-900 [Non-Patent Document 15] Ostrem, JM, et al., K-Ras(G12C) inhibitors allosterically control GTP affinity and effector interactions.Nature 2013,503(7477),548-51 [Non-Patent Document 16] Rudolph, J., et al.,Selective inhibition of mutant Ras protein through covalent binding.Angew Chem Int Ed Engl 2014,53(15),3777-9 [Non-Patent Document 17] Ostrem, JM, et al., Direct small-molecule inhibitors of KRAS: from structural insights to mechanism-based design. Nat Rev Drug Discov 2016, 15(11), 771-785 [Non-Patent Document 18] Nnadi,CI,et al.,Novel K-Ras G12C Switch-II Covalent Binders Destabilize Ras and Accelerate Nucleotide Exchange.J Chem Inf Model 2018,58(2),464-471 [Non-Patent Document 19] Ryan,MBet al.,Vertical Pathway Inhibition Overcomes Adaptive Feedback Resistance to KRAS(G12C)Inhibition.Clin Cancer Res 2019 [Non-Patent Document 20] Xue,JY,et al.,Rapid non-uniform adaptation to conformation-specific KRAS(G12C)inhibition.Nature 2020,577(7790),421-425 [Overview of the Initiative]

[0014] This disclosure describes heterobifunctional compounds that function to recruit Karsten Ras sarcoma protein (KRas or KRAS), e.g., mutant or gain-of-function KRas, to an E3 ubiquitin ligase for targeted ubiquitination and subsequent proteasomal degradation, as well as methods for constructing and using the same. Furthermore, this specification provides methods for using effective amounts of the compounds of this disclosure for the treatment or improvement of disease conditions, e.g., KRas-related diseases or disorders, e.g., accumulation or hyperactivity of KRas protein, or mutant or gain-of-function KRas protein or misfolded KRas protein, 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.

[0015] Thus, in one embodiment, the present disclosure provides a heterobifunctional compound comprising an E3 ubiquitin ligase binding site (i.e., a ligand for E3 ubiquitin ligase ("ULM" group)) and a site that binds to KRas or a variant thereof (i.e., a protein targeting site or "PTM" group, i.e., a KRas targeting ligand or "KTM" group), thereby positioning the KRas protein in close proximity to the ubiquitin ligase, resulting in ubiquitination and subsequent degradation (and / or inhibition) of the KRas protein. In a preferred embodiment, the ULM (ubiquitination ligase binding site) is the von Hippel-Lindou (VHL) E3 ubiquitin ligase binding site (VLM). For example, the structure of the bifunctional compound is: [ka] It can be shown as follows.

[0016] The positions of the PTM and ULM sites (e.g., VLM), and their respective numbers as shown herein, are provided as examples only and are not intended to limit the compounds in any way. As will be understood by those skilled in the art, the bifunctional compounds described herein can be synthesized such that the number and position of each functional site can be varied as desired.

[0017] In a given embodiment, the bifunctional compound further comprises a chemical linker ("L"). In this example, the structure of the bifunctional compound is: [ka] (In the formula, PTM is the KRas targeting site (KTM), L is a linker, e.g., a binding or chemical linking group that binds PTM to ULM, and ULM is the VHL E3 ubiquitin ligase binding site (VLM)).

[0018] For example, the structure of a bifunctional compound is: [ka] (In the formula, PTM may be represented as the KRas targeting site (KTM); "L" is a linker (e.g., a binding or chemical linking group) that binds PTM and VLM; and VLM is a VHL E3 ubiquitin ligase binding site that binds to VHL E3 ubiquitin ligase.)

[0019] In certain embodiments, the compounds described herein include a plurality of independently selected ULMs, a plurality of PTMs, a plurality of chemical linkers, or a combination thereof.

[0020] In any aspect or embodiment described herein, PTM is a small molecule that binds to KRas or its variants, for example, gain-of-function KRas. In any aspect or embodiment described herein, PTM is a small molecule that binds to KRas. In any aspect or embodiment described herein, PTM is a small molecule that binds to both KRas wild-type protein and KRas variants, for example, KRas proteins having gain-of-function mutations. In any aspect or embodiment described herein, PTM is a small molecule that binds to both KRas wild-type protein and KRas variants, for example, gain-of-function KRas variants, but not limited to. In any aspect or embodiment described herein, the small molecule that binds to KRas is as described herein.

[0021] In embodiments, the VLM is a derivative of trans-3-hydroxyproline, where the nitrogen and carboxylic acid in trans-3-hydroxyproline are functionalized as amides. Other intended VLMs are described in U.S. Patent Application Publication 2016 / 0272639 and U.S. Patent Application Publication 2014 / 0356322 (each of which is incorporated herein by reference in whole).

[0022] In a given embodiment, "L" is a linkage. In an additional embodiment, the linker "L" is a conjugate having a linear number of nonhydrogen atoms in the range of 1 to 40 (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, or 40). The conjugate "L" may contain one or more functional groups such as ethers, amides, alkanes, alkenes, alkynes, ketones, hydroxyls, carboxylic acids, thioethers, sulfoxides, and sulfones. The linker may contain aromatic, heteroaromatic, cyclic, bicyclic, or tricyclic moieties. Substitutions with halogens, such as Cl, F, Br, and I, or alkyls, such as methyl, ethyl, isopropyl, and tert-butyl, may be included in the linker. In the case of fluorine substitution, one or more fluorines may be included.

[0023] In additional embodiments, the Specified herein provides compositions comprising an effective amount of a compound described herein or a salt form thereof, and a pharmaceutically acceptable carrier. The therapeutic compositions may be used to induce targeted degradation and / or inhibition of KRas or a variant thereof in a patient or subject, e.g., an animal such as a human, to treat or improve one or more disease conditions, pathologies, or symptoms causally related to KRas or a variant thereof, the treatment being achieved through degradation or inhibition of the KRas protein or a variant thereof in the patient or subject, or through control or reduction of KRas protein levels or protein levels of the variant thereof. In certain embodiments, the therapeutic compositions described herein may be used to achieve the degradation of KRas, or its variants or misfolded forms, for the treatment or improvement of diseases such as the accumulation, aggregation, or hyperreactivity of KRas protein, its misfolded or mutated forms (e.g., gain-of-function KRas protein, 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).

[0024] In yet another aspect, the present disclosure provides a method for ubiquitinating KRas or a variant form in cells (e.g., in vitro or in vivo). In any aspect or embodiment described herein, the method comprises administering a hetero-bifunctional compound described herein, comprising a PTM bound to KRas or a variant form, and preferably a VLM linked via a chemical linker site described herein, in order to achieve degradation of the KRas protein or a variant form. While not wishing to be limited by theory, the inventors believe that according to the present invention, polyubiquitination of KRas wild-type or variant protein occurs when it is positioned near an E3 ubiquitin ligase via the use of a hetero-bifunctional compound, thereby causing degradation of the KRas or variant protein via the proteasome pathway and control or reduction of KRas protein levels in cells (such as target cells requiring such treatment). The control or reduction of the levels of KRas protein or its variant forms provided by this disclosure provides treatment for KRas-related disease conditions, pathologies, or associated symptoms, which are regulated by reducing the amount of KRas protein or its variant forms in the cells of interest.

[0025] Still in another aspect, the Specified provides a method for treating or improving a disease, condition, or symptom thereof in a subject or patient, such as an animal such as a human, that is causally related to KRas or a variant thereof, comprising administering to a subject in need a composition comprising an effective amount, for example, a therapeutically effective amount, of a hetero-bifunctional compound or a salt form thereof as described herein, and a pharmaceutically acceptable carrier, wherein the composition is effective in treating or improving the disease or disorder or symptom thereof in the subject.

[0026] In any embodiment or example described herein, the method involves administering a mutant KRas protein (e.g., KRas) to a target subject before administering the composition or compound of the Disclosure.G12C This further includes identifying the patient as having ).

[0027] In another aspect, this specification provides a method for identifying the effects of KRas protein degradation in biological systems using the compounds of this disclosure.

[0028] In another aspect, the Specified herein provides processes and intermediates for producing heterobifunctional compounds of the Disclosure that are capable of targeted ubiquitination and degradation of KRas proteins in cells (e.g., in vivo or in vitro).

[0029] The accompanying drawings, incorporated herein and forming part of this specification, illustrating some embodiments of this disclosure, serve to illustrate the principles of this disclosure together with this specification. The drawings are solely for the purpose of illustrating embodiments of this disclosure and should not be construed as limiting this disclosure. Further objects, features and advantages of this disclosure will become apparent from the following detailed description, construed together with the accompanying drawings illustrating exemplary embodiments of this disclosure. [Brief explanation of the drawing]

[0030] [Figure 1]A general explanation of the principle of hetero-bifunctional proteolytic compounds. A. An exemplary hetero-bifunctional proteolytic compound comprises a protein targeting site (PTM; dark shaded square), a ubiquitin ligase binding site (ULM; light shaded triangle), and optionally a linker site (L; black line) that binds the PTM to the ULM. B. illustrates the functional use of the hetero-bifunctional proteolytic compounds described herein (commercially known as PROTAC® proteolytic compounds). Briefly, the ULM (triangle) recognizes and binds to a specific E3 ubiquitin ligase, and the PTM (large square) binds to a target protein, recruits it, and brings it into contact with the E3 ubiquitin ligase. Typically, the E3 ubiquitin ligase is complexed with an E2 ubiquitin conjugate protein (E2) and, either alone or via the E2 protein, catalyzes the binding of multiple ubiquitin molecules (black circles) to lysine on the target protein via isopeptide bonds. Polyubiquitinated proteins (far right) are thus targeted for degradation by the cellular proteasome mechanism. [Figure 2A] The MRTX849-VHL bifunctional compound engages with and degrades endogenous KRASG12C in NCI-H2030 cells: Chemical structures of MRTX849, LC-1, LC-2 (active bifunctional proteolytic compound), and LC-2 epimer. [Figure 2B] The bifunctional compound MRTX849-VHL engages with and degrades endogenous KRASG12C in NCI-H2030 cells: LC-1 engages with KRASG12C in a dose-dependent manner. Quantitative analysis is shown on the right. [Figure 2C] The bifunctional compound MRTX849-VHL engages with and degrades endogenous KRASG12C in NCI-H2030 cells: LC-2 degrades KRASG12C in a dose-dependent manner. The right side shows quantitative analysis. Quantitative data represent mean ± SD. Not significant (NS); *p<0.05; **p<0.01; ****p<0.001. [Figure 3]The docking and LC-2 degradation of MRTX849 are specific to KRASG12C. (A) Docking of MRTX849 (MRTX) to the crystal structure of KRASG12C (PDB: 5V9U). MRTX is shown in yellow, and the black arrows indicate linker junctions. (B) LC-2 does not degrade KRASG12D in HCT116 cells. The right side shows quantitative results. Quantitative data are shown as mean ± SD. [Figure 4A] LC-2 induces KRASG12C degradation in multiple mutant cell lines. LC-2 (but not the LC-2 epimer) induces KRASG12C degradation in heterozygous H358 cells. Quantitative results are shown on the right. [Figure 4B] LC-2 induces KRASG12C degradation in multiple mutant cell lines. LC-2 induces KRASG12C in homozygous MIA PaCa-2 cells. Quantitative results are shown on the right. [Figure 4C] LC-2 induces KRASG12C degradation in multiple mutant cell lines. LC-2 induces KRASG12C degradation in homozygous MRTX849-resistant SW1573 cells. [Figure 4D] LC-2 induces KRASG12C degradation in multiple mutant cell lines. LC-2 induces KRASG12C degradation in heterozygous NCI-H23. The right side shows quantitative results. Quantitative data represent mean ± SD. *p<0.05;**p<0.01;***p<0.005;****p<0.001. [Figure 5A] Endogenous KRASG12C degradation is mediated by heterobifunctional compounds. The LC-2 epimer does not induce KRASG12C degradation at 2.5 μM; LC-2-induced degradation is rescued in NCI-H2030 cells by VHL ligand competition, proteasome inhibition with epixomicin (Epox), and nedilation inhibition with MLN4924 (MLN). Quantitative results are shown below. [Figure 5B]Endogenous KRASG12C degradation is mediated by heterobifunctional compounds. Inhibition of nedilation, rather than inhibition of lysosomal acidification, rescues LC-2-induced KRASG12C degradation in NCI-H23 cells. Quantitative results are shown below. Quantitative data represent mean ± SD. Non-significant (NS); ***p<0.005. [Figure 6] KRASG12C degradation is rapid, with maximal degradation induced as quickly as 4 hours: (A) Time course in NCI-H2030 cells. LC-2 and LC-2 epimers engage within 1 hour, maximal degradation observed by 8 hours, and maintained for up to 24 hours. Quantification is shown on the right. (B) Time course in SW1573 cells. LC-2 and LC-2 epimers engage with KRAS within 1 hour, maximal degradation observed at 12 hours, and maintained for up to 24 hours. Quantification is shown on the right. The LC-2 epimer is a quantification of the high molecular weight bifunctional compound epimer modification band to monitor the engagement of KRASG12C over time, rather than total KRAS levels. Quantitative data represent mean ± SD. Non-significant (NS); *p<0.05; **p<0.01; ***p<0.005; ****p<0.001. [Figure 7] Endogenous KRASG12C degradation is maintained over 72 hours in multiple cancer cell lines. (A) Time course over 72 hours in MIA PaCa-2 cells. Degradation begins at 6 hours and is maintained for a maximum of 72 hours. Quantitative analysis is shown on the right. (B) Time course over 72 hours in NCI-H23 cells. Degradation begins within 6 hours, reaches a maximum at 24 hours, and begins to rebound by 72 hours. Quantitative analysis is shown on the right. Quantitative data represent mean ± SD. Not significant (NS); **p<0.01; ***p<0.005; ****p<0.001 [Figure 8] LC-2-induced KRASG12C degradation is maintained in SW1573 for 72 hours. LC-2-induced KRASG12C occurs within 6 hours and is maintained for 72 hours. No changes are observed in the LC-2 epimer. [Figure 9A]Degradation of endogenous KRASG12C regulates Erk signaling in homozygous and heterozygous KRASG12C cell lines. Degradation of KRASG12C in homozygous NCI-H2030 cells dose-dependently attenuates pErk. (Right image shows quantitative analysis.) [Figure 9B] Endogenous KRASG12C degradation regulates Erk signaling in homozygous and heterozygous KRASG12C cell lines. KRASG12C degradation in heterozygous NCI-H23 cells dose-dependently reduces pErk. Quantitative results are shown on the right. See Tables 2 and 3 for statistical analysis. Quantitative data are expressed as mean ± SD. [Figure 10A] Effects of KRASG12C degradation and inhibition on Erk signaling over time. Inhibition and degradation of KRASG12C reduce pErk signaling at 6 and 24 hours in homozygous MIA PaCa-2 cells. Quantitative results are shown on the right. [Figure 10B] Effects of KRASG12C degradation and inhibition on Erk signaling over time. Inhibition and degradation of KRASG12C reduce pErk signaling at 6 and 24 hours in heterozygous NCI-H23. Quantitative results are shown on the right. See Tables 4 and 5 for statistical analysis. Quantitative data are expressed as mean ± SD. [Figure 11] Changes in Erk signaling in SW1573 cells during 24-hour LC-2 treatment. Erk signaling is regulated by LC-2. pErk decreases throughout the time course. The right side shows quantitative values. Quantitative data represent mean ± SD. See Table 6 for statistical analysis. [Figure 12] The chemical structure, linker length (from the carbon adjacent to the pyrrolidine nitrogen to the carbon adjacent to the VHL carbonyl), and activity are shown for exemplary compounds LC-1, LC-2, LC-3, LC-4, LC-5, and LC-6. Shorter linker lengths induce higher levels of degradation. Data from aNCI-H2030 cells. Data from bSW1573 cells. [Modes for carrying out the invention]

[0031] This disclosure describes compounds, compositions, and methods relating to the remarkable discovery that when an E3 ubiquitin ligase and a KRas protein are positioned in proximity via a bifunctional compound that binds to both the E3 ubiquitin ligase and the KRas protein, the E3 ubiquitin ligase (e.g., von Hippel-Lindou (VHL) E3 ubiquitin ligase) ubiquitinates the KRas protein or its variant forms. Accordingly, this disclosure provides compounds and compositions comprising an E3 ubiquitin ligase binding site ("ULM") bound to or by a chemical linking group (L) to a protein targeting site ("PTM") that targets the KRas protein, resulting in ubiquitination of the KRas protein and subsequent degradation of the KRas protein by the proteasome (see Figures 1A and 1B).

[0032] In some embodiments, this specification provides compounds to which PTMs bind to the KRas protein and / or its variant forms. This disclosure also provides a library of compositions and their use for resulting in targeted degradation of the KRas protein in cells.

[0033] In a given aspect, the Disclosure provides heterobifunctional compounds comprising ligands capable of binding to E3 ubiquitin ligases, such as von Hippel-Lindou E3 ubiquitin ligase, e.g., small molecule ligands (i.e., having molecular weights less than 2,000, 1,000, 500, or 200 Daltons). The compounds also include small molecule sites capable of binding to the KRas protein or its variants in such a manner that the KRas protein or variant is positioned near the ubiquitin ligase, resulting in ubiquitination and degradation (and / or inhibition) of the KRas protein or variant. "Small molecule" means, in addition to the above, that the molecule is non-peptidyl, i.e., it is not considered a peptide and contains, for example, fewer than 4, 3, or 2 amino acid residues. According to this Specification, each of the PTM, ULM, and heterobifunctional molecules is a small molecule.

[0034] When the term "KRas" is used throughout this specification, unless otherwise specifically indicated, it is intended to include wild-type KRas and its variant forms, such as gain-of-function KRas variant proteins or KRas proteins having codon 12 missense mutations, codon 12 missense mutations, exon 2 mutations, G12V, G12C, G12D, G12A, G13D, exon 3 mutations, codon 61 missense mutations, exon 4 mutations, G12R, Q61H, G12S, A146T, G13C, Q61R, Q61L, A146V, codon 117 missense mutations, K117N, Q61K, G12F, codon 59 missense mutations, A59T, or a combination thereof or one or more mutations selected from such combinations.

[0035] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art in which this disclosure pertains. The technical terms used in this specification are solely for the purpose of describing specific embodiments and are not intended to limit this disclosure.

[0036] Where a range of values ​​is provided, unless the context explicitly indicates otherwise, it is understood that each intervening value within that range, up to one-tenth of the lower limit (for example, in the case of a group containing many carbon atoms where the number of carbon atoms each falls within the range is provided), and any other described or intervening values ​​within that described range are included in this disclosure. The upper and lower limits of these smaller ranges are independently included within those smaller ranges and may also be included in this disclosure, depending on any particularly excluded limits within the described range. Where a described range includes one or both of the limiting values, the range excluding one or both of those included limiting values ​​is also included in this disclosure.

[0037] The following terms are used to describe this disclosure. If a term is not specifically defined herein, it is given the meaning that will be recognized in the art by a person skilled in the art as applied in the context of its use in describing this disclosure.

[0038] The articles "a" and "an," when used herein and in the appended claims, refer to one or more (i.e., at least one) grammatical objects of the article unless otherwise explicitly indicated by the context. For example, "an element" means one or more elements unless otherwise indicated.

[0039] In the claims and in the specification above, all transitional phrases such as “comprising,” “including,” “possessing,” “having,” “containing,” “involving,” “holding,” and “composed of” should be understood to be open-ended, meaning that they include but are not limited to. Only the transitional phrases “consisting of” and “essentially consisting of” shall be closed or semi-closed, respectively, as provided in Section 2111.03 of the U.S. Patent and Trademark Office Manual for Patent Examination Procedure.

[0040] In any method or process described herein that includes multiple steps or actions, it should be understood that, unless otherwise indicated by the context, the order of the steps or actions of the method is not necessarily limited to the order in which the steps or actions of the method are described.

[0041] The terms “co-administration” and “co-administration” or “combination therapy” refer to both co-administration (simultaneous administration of two or more therapeutic agents) and time-dependent administration (administration of one or more therapeutic agents at different times than the administration of additional therapeutic agents or drugs), as long as two or more therapeutic agents are present in the patient to some extent, preferably in effective amounts, at the same time. In certain preferred embodiments, one or more of the heterobifunctional compounds described herein are co-administered with at least one additional bioactive agent, such as an anticancer agent. In particularly preferred embodiments, co-administration of such compounds results in synergistic activity and / or therapy, such as anticancer activity.

[0042] The term “compound,” as used herein, means any specific heterodifunctional compound disclosed herein, its pharmaceutically acceptable salts and solvates, and any deuterated form (where applicable) of any of the aforementioned molecules, unless otherwise specified. A deuterated compound is one in which one or more hydrogen atoms in the drug molecule are replaced with deuterium. Such a deuterated compound preferably has one or more improved pharmacokinetic or pharmacodynamic properties (e.g., a longer half-life) compared to an equivalent “non-deuterated” compound.

[0043] The term "ubiquitin ligase" refers to a family of proteins that facilitate the transfer of one or more ubiquitins to a specific substrate protein. The addition of several ubiquitin chains (polyubiquitination) targets the substrate protein for degradation. For example, von Hippel-Lindou is an E3 ubiquitin ligase that, alone or in combination with E2 ubiquitin conjugates, ultimately causes the binding of four ubiquitin chains to a lysine residue on a target protein, thereby targeting that protein for degradation by the proteasome. Ubiquitin ligases are involved in polyubiquitination such that the first ubiquitin binds to the lysine on the target protein; the second ubiquitin binds to the first; the third ubiquitin binds to the second; and the fourth ubiquitin binds to the third. Such polyubiquitination marks the protein for degradation by the proteasome.

[0044] The terms “patient” or “subject” are used throughout this specification to describe animals, preferably humans or domesticated animals, to which treatment, including preventive measures, is provided with the compositions of this disclosure. For the treatment of diseases, conditions or symptoms specific to certain animals, such as human patients, the term “patient” refers to that particular animal, including domesticated animals, such as dogs or cats, or farm animals, such as horses, cattle, or sheep. Generally in this disclosure, the terms “patient” and “subject” refer to human patients unless otherwise stated or implied by the context of their use.

[0045] The terms “effective” and “therapeutically effective,” when used in the context of their intended use, are used to describe the amount of a compound or composition that, in a single dose or more preferably after multiple doses in the context of a treatment regimen, results in an intended outcome, such as improvement of a disease or condition, or improvement or reduction of one or more symptoms associated with a disease or condition. The terms “effective” and “therapeutically effective” encompass all other terms “effective amount” or “effective concentration” as otherwise described or used in this application.

[0046] Compounds and compositions

[0047] In one embodiment, this specification provides a heterobifunctional compound comprising an E3 ubiquitin ligase binding site ("ULM") which is a VHL E3 ubiquitin ligase binding site ("VLM"). The VLM is covalently bound to a protein targeting site (PTM) that binds to a protein, and the binding is direct by binding or structural: (A) PTM-L-VLM The binding occurs via a chemical linking group (L) by the KRas targeting site (KTM) (wherein L is a binding or chemical linking group, PTM is a protein targeting site that binds to the protein KRas or a variant form described herein, and PTM is the KRas targeting site (KTM)). The term VLM encompasses all VHL binding sites.

[0048] In any aspect or embodiment, VLM is used to achieve a maximum half-mass inhibitory concentration (IC) for E3 ubiquitin ligases less than approximately 200 μM (e.g., VHL E3 ubiquitin ligase). 50 ) demonstrates IC 50 This can be determined according to any preferred method known in the art, for example, a fluorescence polarization assay.

[0049] In certain embodiments, the hetero-bifunctional compounds described herein have an IC 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 50 or demonstrate a maximum half-decomposition concentration (DC 50 ).

[0050] In any aspect or embodiment described herein, the PTM has a chemical structure: [Chemical Formula] (wherein of PTM [Chemical Formula] is a binding site for VLM or L that binds VLM to PTM; [Chemical Formula] is a 6-membered aryl, 6-membered heteroaryl, or 6-membered heterocycloalkyl, each optionally substituted with 1 or 2 halogens (e.g., Cl, F, or Br); R PTM2 is -C(=O)C2-C4 alkenyl optionally substituted by methyl or halogen (e.g., Cl, F, Br); R PTM3A is H, phenyl, or naphthalene, each optionally substituted by 1, 2, or 3 groups independently selected from OH, halogen (e.g., F, Cl, Br), or linear or branched C1-C3 alkyl (e.g., methyl or ethyl); R PTM3B is H, halogen (e.g., Cl, F, Br), or -O-R PTM3C and RPTM3C is indazole (for example, [ka] ) and R PTM3B It is optionally substituted with one, two, or three groups independently selected from OH, halogens (e.g., F, Cl, Br), or linear or branched C1-C3 alkyl groups (e.g., methyl or ethyl); R PTM4A is one or two independently selected halogens (e.g., Cl, F, Br); R PTM4B (1) -CH2-CH2-CN or -CH2-CN, or (2) one or two independently selected C1-C3 alkyl groups (e.g., methyl or ethyl); each X PTM (Each is either CH or N) It is represented by [this].

[0051] The term "alkyl" in this context refers to a linear, branched, or cyclic fully saturated hydrocarbon radical, preferably C1-C1. 10 , preferably C1-C6, or more preferably C1-C3 alkyl groups (which may be optionally substituted with any one or more suitable functional groups). Examples of alkyl groups include, among others, 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. In certain embodiments, the alkyl group is end-capped with a halogen group (At, Br, Cl, F, or I).

[0052] The term "alkenyl" refers to a linear, branched, or cyclic C2-C bond containing at least one C=C bond. 10 (Preferably C2-C6) hydrocarbon groups are referred to.

[0053] The term "alkynyl" refers to a linear, branched, or cyclic C2-C bond containing at least one C≡C bond. 10 (Preferably C2-C6) hydrocarbon groups are referred to.

[0054] The term "alkylene" may be optionally substituted when used - (CH2) n - A group (n is usually an integer from 0 to 6) is used. If substituted, the alkylene group is preferably substituted with one or more methylene groups, with a C1-C6 alkyl group (including a cyclopropyl group or a t-butyl group), but may be substituted with one or more halo groups, preferably 1 to 3 halo groups or 1 or 2 hydroxyl groups, O-(C1-C6 alkyl) groups or amino acid side chains as otherwise disclosed herein. In certain embodiments, the alkylene group may be substituted with a urethane or alkoxy group (or other preferred functional group), and the urethane or alkoxy group (or other preferred group) may be further substituted with a polyethylene glycol chain (of 1 to 10, preferably 1 to 6, or more preferably 1 to 4 ethylene glycol units), which is substituted with an alkyl chain (preferably, but not exclusively, at the distal end of the polyethylene glycol chain) substituted with one halogen group, preferably a chlorine group. In further embodiments, the alkylene (e.g., methylene) group may be substituted with an amino acid side chain group, for example, a side chain group of a natural or unnatural amino acid, such as alanine, β-alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan, or tyrosine.

[0055] The term "unsubstituted" means that only hydrogen atoms are substituted. The range of carbon atoms including C0 means that carbon is absent and replaced by H. Therefore, the range of carbon atoms C0-C6 includes 1, 2, 3, 4, 5, and 6 carbon atoms, and in the case of C0, H is present in place of carbon.

[0056] The terms “substituted” or “optionally substituted” mean, independently (i.e., if multiple substituents exist, each substituent is selected independently of the other substituents) one or more substituents at any carbon (or nitrogen) position on any site on the molecule in the context (independently, up to five substituents, preferably up to three substituents, more preferably one or two substituents (which may include substituents that themselves may be further substituted) on the site in the compounds of this disclosure), where possible substituents: hydroxyl, thiol, carboxyl, cyano (C≡N), nitro (NO₂), halogen (preferably, in particular alkyl, especially one, two or three halogens on a methyl group, e.g., trifluoromethyl), alkyl (preferably C1-C 10, more preferably C1-C6), aryl (particularly phenyl and substituted phenyl, e.g., benzyl or benzoyl), alkoxy group (preferably C1-C6 alkyl or aryl, including phenyl and substituted phenyl), thioether (preferably C1-C6 alkyl or aryl), acyl (preferably C1-C6 acyl), alkylene ester (preferably substituted with a C1-C6 alkyl or aryl group, such that the bond is on the alkylene group rather than the ester functional group), ester or thioester (preferably C1-C6 alkyl or aryl), halogen (preferably F or Cl), amine (including 5 or 6-membered cyclic alkyleneamine, C1-C6 alkylamine) The material comprises a C1-C6 dialkylamine (which may further include an alkyl group that is substituted with one or two hydroxyl groups) or an optionally substituted -N(C0-C6 alkyl)C(O)(O-C1-C6 alkyl) group (which may further be substituted with a polyethylene glycol chain bonded to an alkyl group that contains a single halogen, preferably a chlorine substituent), hydrazine, amide (preferably independently substituted with one or two C1-C6 alkyl groups) (including carboxamides optionally substituted with one or two C1-C6 alkyl groups), alkanol (preferably C1-C6 alkyl or aryl), or alkanolic acid (preferably C1-C6 alkyl or aryl). Substituents according to this disclosure may include, for example, a -SiR1R2R3 group (wherein R1 and R2 are as otherwise described herein, and R3 is H or a C1-C6 alkyl group, preferably R1, R2, and R3 together are a C1-C3 alkyl group (including an isopropyl or t-butyl group)). Each of the above groups may be directly linked to the substitution site, or alternatively, the substituent may be optionally substituted -(CH2) m -or optionally substituted with -(OCH2) m -,-(OCH2CH2) m -or-(CH2CH2O) m- A group (which may be substituted with any one or more of the substituents described above) can be linked to the substitution site (preferably an aryl or heteroaryl site). Alkylene group-(CH2) m -or-(CH2) n - The group or other chain, for example, the ethylene glycol chain (specified above), may be substituted at any point on the chain. Preferred substituents on the alkylene group include halogens or C1-C6 (preferably C1-C3) alkyl groups (which may be optionally substituted with one or two hydroxyl groups, one or two ether groups (O-C1-C6 groups), up to three halo groups (preferably F), or amino acid side chains as otherwise described herein) and optionally substituted amides (preferably substituted carboxamides as described above) or urethane groups (which often have one or two C0-C6 alkyl substituents, the group(s) of which may be further substituted). In certain embodiments, the alkylene group (often a single methylene group) is substituted with one or two optionally substituted C1-C6 alkyl groups, preferably C1-C4 alkyl groups, most often methyl or O-methyl groups, or amino acid side chains as otherwise described herein. In this disclosure, a site in a molecule may be optionally substituted with up to five substituents, preferably up to three substituents. Most often in this disclosure, the substituted site is substituted with one or two substituents.

[0057] The term “substituted” (each substituent is independent of any other substituent) also means, in the context of its use, C1-C6 alkyl, C1-C6 alkoxy, halogen, amide, carboxamide, sulfone, keto, carboxy, C1-C6 ester (oxyester or carbonyl ester), C1-C6 keto, urethane-OC(O)-NR1R2 or -N(R1)-C(O)-O-R1, nitro, cyano and amine (in particular C1-C6 alkylene-NR1R2, mono or di-C1-C6 alkyl-substituted amines (which may optionally be substituted with one or two hydroxyl groups)). Each of these groups, in the context unless otherwise indicated, contains 1 to 6 carbon atoms. In a given embodiment, preferred substituents are, depending on the context of the substituent’s use, e.g., -NH-, -NHC(O)-, -O-, =O, -(CH2) m -(where m and n are 1, 2, 3, 4, 5, or 6 in the context), -S-, -S(O)-, SO2- or -NH-C(O)-NH-, -(CH2) n OH, -(CH2) n SH, -(CH2) n COOH, C1-C6 alkyl, -(CH2) n O-(C1-C6 alkyl), -(CH2) n C(O)-(C1-C6 alkyl),-(CH2) n OC(O)-(C1-C6 alkyl),-(CH2) n C(O)O-(C1-C6 alkyl),-(CH2) n NHC(O)-R1, -(CH2) n C(O)-NR1R2, -(OCH2) n OH, -(CH2O) n COOH, C1-C6 alkyl, -(OCH2) n O-(C1-C6 alkyl), -(CH2O) n C(O)-(C1-C6 alkyl),-(OCH2) n NHC(O)-R1, -(CH2O) n C(O)-NR1R2, -S(O)2-R S ,-S(O)-R S (R SC1-C6 alkyl or -(CH2) m The alkylene group may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein, but may also be substituted as otherwise disclosed herein Various arbitrarily substituted sites may be substituted with three or more substituents, preferably three or fewer substituents, and preferably one or two substituents. It should be noted that in compounds at a particular position on the molecule, substitution is required (mainly by valency), but if substitution is not indicated, then that substituent is interpreted or understood to be H unless otherwise suggested by the context of substitution.

[0058] The terms “aryl” or “aromatic” in this context refer to a substituted (as otherwise described herein) or unsubstituted monovalent aromatic group (e.g., a 5- to 16-membered ring) having a single ring (e.g., benzene, phenyl, benzyl, or a 5, 6, 7, or 8-membered ring) or a fused ring (e.g., naphthyl, anthracenyl, phenantrenyl, or a 10- to 16-membered ring), which may be attached to the compounds of this disclosure at any available stable position on the ring(s) or as otherwise shown in the presented chemical structures. Other examples of aryl groups, in context, may include, among others, heterocyclic aromatic ring systems, "heteroaryl" groups having one or more nitrogen, oxygen, or sulfur atoms in the ring (monocyclic), such as imidazole, furyl, pyrrole, furanyl, thien, thiazole, pyridine, pyrimidine, pyrazine, triazole, oxazole, or fused ring systems, such as indole, quinoline, indidine, azaindridine, benzofurazan, etc. (these may be optionally substituted as described above).Among the possible heteroaryl groups, in particular nitrogen-containing heteroaryl groups, such as pyrrole, pyridine, pyridone, pyridazine, pyrimidine, pyrazine, pyrazole, imidazole, triazole, triazine, tetrazole, indole, isoindole, indidine, azaindidine, purine, indazole, quinoline, dihydroquinoline, tetrahydroquinoline, isoquinoline, dihydroisoquinoline, tetrahydroisoquinoline, quinoridine, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, imidazopyridine, imidazotriazine, pyrazinopyridazine, acridine, phenanthridine, carbazole, carbazoline, pyrimidine, phenanthroline, phenacene, oxadiazole, benzyl The materials include midazoles, pyrrolopyridines, pyrrolopyrimidines, and pyridopyrimidines; sulfur-containing aromatic heterocycles, e.g., thiophenes and benzothiophenes; oxygen-containing aromatic heterocycles, e.g., furans, pyrans, cyclopentapyrans, benzofurans, and isobenzofurans; and aromatic heterocycles containing two or more heteroatoms selected from nitrogen, sulfur, and oxygen, e.g., thiazoles, thiadizols, isothiazoles, benzoxazoles, benzothiazoles, benzothiadiazoles, phenothiazines, isoxazoles, furazans, phenoxazines, pyrazoloxazoles, imidazothiazoles, thienofurans, phlopyrroles, pyridoxazines, phlopyridines, phlopyrimidines, thienofyrimidines, and oxazoles, all of which may be optionally substituted.

[0059] The term "substituted aryl" refers to an aromatic carbocyclic group comprising at least one aromatic ring or multiple fused rings (at least one of which is aromatic), where one or more rings are substituted with one or more substituents. For example, an aryl group is -(CH2) n OH, -(CH2) n -O-(C1-C6)alkyl, -(CH2) n -O-(CH2) n -(C1-C6) alkyl, -(CH2) n -C(O)(C0-C6)alkyl, -(CH2) n-C(O)O(C0-C6)alkyl, -(CH2) n -OC(O)(C0-C6) alkyl, amine, mono or di(C1-C6 alkyl)amine (wherein the alkyl group on the amine is optionally substituted with one or two hydroxyl groups or up to three halo (preferably F, Cl) groups), OH, COOH, C1-C6 alkyl, preferably CH3, CF3, OMe, OCF3, NO2, or CN group (each of these may be substituted at the ortho, meta and / or para position of the phenyl ring, preferably para), optionally substituted phenyl group (phenyl group The compound itself is preferably connected to a PTM containing a ULM group via a linker group), and / or F, Cl, OH, COOH, CH3, CF3, OMe, OCF3, NO2, or CN group (ortho, meta and / or para positions of the phenyl ring, preferably para), optionally substituted naphthyl group, optionally substituted heteroaryl, preferably optionally substituted isoxazole including methyl-substituted isoxazole, optionally substituted oxazole including methyl-substituted oxazole, and methyl-substituted thiazole. It may contain at least one of the following: optionally substituted thiazoles, optionally substituted isothiazoles including methyl-substituted isothiazoles, optionally substituted pyrroles including methyl-substituted pyrroles, optionally substituted imidazoles including methylimidazoles, optionally substituted benzimidazoles or methoxybenzylimidazoles, optionally substituted oxymidazoles or methyloxymidazoles, optionally substituted diazole groups including methyldiazole groups, optionally substituted triazole groups including methyl-substituted triazole groups, optionally substituted pyridine groups including halo-(preferably F) or methyl-substituted pyridine or oxapyridine groups (the pyridine group is linked to the phenyl group by oxygen), optionally substituted furans, optionally substituted benzofurans, optionally substituted dihydrobenzofurans, optionally substituted indoles, indolindine or azaindridine (2, 3, or 4-azandridine), optionally substituted quinolines, and substituents(s) selected from combinations thereof.

[0060] "Carboxyl" is represented by --C(O)OR (wherein R is hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, or substituted heteroaryl, while these comprehensive substituents have the same meaning as the definition of the corresponding group as defined herein).

[0061] The term "hetalial" or "hetalial" refers to 5- to 16-membered heteroaryls (e.g., 10- to 16-membered heteroaryls having a 5, 6, 7, or 8-membered monocyclic ring or multiple fused rings), optionally substituted quinolines (which may be bonded to a pharmacophore or substituted on any carbon atom within the quinoline ring), optionally substituted indoles (including dihydroindole), optionally substituted indridines, optionally substituted azaindridines (2, 3, or 4-azaidinidine), optionally substituted benzimidazoles, and benzodiazos. Electrolytes, benzoxofran, optionally substituted imidazole, optionally substituted isoxazole, optionally substituted oxazole (preferably methyl-substituted), optionally substituted diazole, optionally substituted triazole, tetrazole, optionally substituted benzofuran, optionally substituted thiophene, optionally substituted thiazole (preferably methyl and / or thiol-substituted), optionally substituted isothiazole, optionally substituted triazole (preferably methyl group, triisopropylsilyl group, optionally substituted -(CH2) m -O-C1-C6 alkyl group or optionally substituted -(CH2) m -C(O)-O-C1-C6 alkyl group substituted 1,2,3-triazoles), optionally substituted pyridines (2-,3, or 4-pyridines), or chemical structures: [ka] (In the formula, S c CHR SS , NR URE , or O; R HETThis can be H, CN, NO2, halo (preferably Cl or F), optionally substituted C1-C6 alkyl (preferably substituted with 1 or 2 hydroxyl groups or up to 3 halo groups (e.g., CF3)), optionally substituted O(C1-C6 alkyl) (preferably substituted with 1 or 2 hydroxyl groups or up to 3 halo groups), or optionally substituted acetylene group-C≡CR a (In the formula, R a (is H or a C1-C6 alkyl group (preferably a C1-C3 alkyl group); R SS is H, CN, NO2, halo (preferably F or Cl), optionally substituted C1-C6 alkyl (preferably substituted with 1 or 2 hydroxyl groups or up to 3 halo groups), optionally substituted O-(C1-C6 alkyl) (preferably substituted with 1 or 2 hydroxyl groups or up to 3 halo groups), or optionally substituted -C(O)(C1-C6 alkyl) (preferably substituted with 1 or 2 hydroxyl groups or up to 3 halo groups); R URE is H, C1-C6 alkyl (preferably H or C1-C3 alkyl) or -C(O)(C1-C6 alkyl) (each of which is optionally substituted with one or two hydroxyl groups or up to three halogens, preferably fluorine groups), or optionally substituted heterocycles, such as piperidine, morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, piperidine, piperazine (each of which is optionally substituted), Y C is N or CR YC (In the formula, R YC This can be H, OH, CN, NO2, halo (preferably Cl or F), optionally substituted C1-C6 alkyl (preferably substituted with 1 or 2 hydroxyl groups or up to 3 halo groups (e.g., CF3)), optionally substituted O(C1-C6 alkyl) (preferably substituted with 1 or 2 hydroxyl groups or up to 3 halo groups), or optionally substituted acetylene group-C≡CR a (In the formula, Ra (This is H or a C1-C6 alkyl group (preferably a C1-C3 alkyl group)) It may mean a basis by, but is not limited to, these.

[0062] The terms "aralkyl" and "heteroarylalkyl" refer to groups that include both aryl and heteroaryl groups, respectively, as well as alkyl and / or heteroalkyl and / or carbocyclic and / or heterocycloalkyl ring systems as defined above.

[0063] The term "arylalkyl," as used herein, refers to the aryl group defined above attached to the alkyl group defined above. An arylalkyl group is bonded to a parent site via an alkyl group, and the alkyl group consists of 1 to 6 carbon atoms. The aryl group in an arylalkyl group may be substituted as defined above.

[0064] The term "heterocyclic" refers to a cyclic group containing at least one heteroatom, e.g., N, O, or S, and which may be aromatic (heteroaryl) or non-aromatic. Therefore, heteroaryl moieties are included under the definition of heterocyclic, depending on the context of their use. Exemplary heteroaryl groups are described above herein.

[0065] Exemplary heterocycles include, in particular, azetidinyl, benzimidazolyl, 1,4-benzodioxanyl, 1,3-benzodioxolyl, benzoxazolyl, benzothiazolyl, benzothienyl, dihydroimidazolyl, dihydropyranyl, dihydrofuranyl, dioxanyl, dioxolanyl, ethyleneurea, 1,3-dioxolane, 1,3-dioxane, 1,4-dioxane, furyl, homopiperidinyl, imidazolyl, imidazolinyl, imidazolidinyl, indolinyl, indolyl, isoquinolinyl, isothiazolidinyl, isothiazolyl, isoqui This includes sazolidinil, isoxazolyl, morpholinil, naphthilidinil, oxazolidinil, oxazolyl, pyridone, 2-pyrrolidone, pyridine, piperazinil, N-methylpiperazinil, piperidinil, phthalimide, succinimide, pyrazinil, pyrazolinil, pyridyl, pyrimidinil, pyrrolidinil, pyrrolinil, pyrrolyl, quinolinil, tetrahydrofuranil, tetrahydrothiopyranil, tetrahydroquinoline, thiazolidinil, thiazolyl, thienyl, tetrahydrothiophene, oxane, oxetanil, oxathiolanil, and thian.

[0066] The heterocyclic group may be optionally substituted with members selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azide, cyano, halogen, hydroxyl, keto, thioketo, carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, oxo (=O), and -SO2-heteroaryl. Such a heterocyclic group may have a single ring or multiple fused rings. Examples of nitrogen heterocycles and heteroaryls include, but are not limited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indidine, isoindole, indole, indazole, purine, quinolidine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carborin, phenantholidine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, morpholino, piperidinyl, tetrahydrofuranil, and N-alkoxy-nitrogen-containing heterocycles. The term "heterocyclic" also includes bicyclic groups in which any of the heterocyclic rings is condensed with a benzene ring, a cyclohexane ring, or another heterocyclic ring (e.g., indolyl, quinolyl, isoquinolyl, tetrahydroquinolyl, etc.).

[0067] The term "cycloalkyl" means, but is not limited to, monocyclic or polycyclic alkyl groups or cycloalkanes as defined herein, including, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc., a monovalent group derived from a saturated monocyclic hydrocarbon group having 3 to 20 carbon atoms in the ring, for example, but is not limited to these. The term "substituted cycloalkyl" means, but is not limited to, a monocyclic or polycyclic alkyl group substituted with one or more substituents, for example, amino, halogen, alkyl, substituted alkyl, carbyloxy, carbyl mercapto, aryl, nitro, mercapto, or sulfo, while these comprehensive substituents have the same meaning as the definitions of the corresponding groups as defined in this legend.

[0068] "Hypercycloalkyl" 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 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, carbyl mercapto, aryl, nitro, mercapto, or sulfo, while these comprehensive substituents have the same meaning as the definitions of the corresponding groups as defined in this legend.

[0069] The term "hydrocarbyl" refers to a compound containing carbon and hydrogen, which may be fully saturated, partially unsaturated, or aromatic, and includes aryl, alkyl, alkenyl, and alkynyl groups.

[0070] The term "independently" is used herein to indicate that a variable element that is applied independently changes independently with each application.

[0071] The term "lower alkyl" refers to methyl, ethyl, or propyl alkyl groups.

[0072] The term "lower alkoxy" refers to methoxy, ethoxy, or propoxy compounds.

[0073] Exemplary VLM

[0074] In any aspect or embodiment described herein, ULM is VLM, and its chemical structure is: [ka] (In the formula, R 14 In any embodiment or configuration described herein, R 14 , R 14a , or R 14b As defined in; R 15 This is defined as in any embodiment or example described herein; R 16 This is defined as in any embodiment or example described herein; o is as defined in any aspect or embodiment described herein; [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.) It is represented by [this].

[0075] For example, in any aspect or embodiment described herein, ULM is VLM, and its chemical structure is: [ka] (In the formula, R 14 is H or linear or branched C1-C3 alkyl (e.g., methyl); R 15 This is a 5-membered heteroaryl having one or two heteroatoms selected from N, S, and O, optionally substituted with CN or methyl (e.g., [ka] ) and; R 16 is a halo, optionally substituted C1-C3 alkyl, optionally substituted C1-C3 haloalkyl, hydroxy, optionally substituted C1-C3 alkoxy, or optionally substituted C1-C3 haloalkoxy; o is an integer between 0 and 2 (for example, 0, 1, or 2); [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.) It is represented by [this].

[0076] In any aspect or embodiment described herein, ULM is VLM, and is based on ULM-a: [ka] (In the formula, [ka] This indicates the bonding of a chemical linker site that bonds at least one PTM, another ULM or VLM (i.e., ULM' or VLM'), or at least one PTM, ULM' or VLM' to the other end of the linker; X of formula ULM-a 1 , X 2 is bond, O, NR Y3 , CR Y3 R Y4 , independently selected from the groups C=O, C=S, SO, and SO2; R of formula ULM-a Y3 , RY4 is independently selected from the group of H, linear or branched C optionally substituted by one or more halos 1-6 alkyl, optionally substituted C 1-6 alkoxyl (for example, optionally substituted by 0 to 3 R P groups); R in formula ULM-a P is 0, 1, 2, or 3 groups independently selected from H, halo, -OH, C 1-3 alkyl, C=O; W in formula ULM-a 3 is optionally substituted T, optionally substituted -T-N(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-bisheteroaryl, optionally substituted -T-heterocycle, optionally substituted -T-bisheterocycle, optionally substituted -NR 1 -T-aryl, optionally substituted -NR 1 -T-heteroaryl, or optionally substituted -NR 1 -T-heterocycle selected from the group; X in formula ULM-a 3 is C=O, R 1 , R 1a , R 1b ; R 1 , R 1a , R 1b each of which is H, a linear or branched C1-C6 alkyl group optionally substituted by one or more halo or -OH groups, R Y3 C=O, R Y3 C=S, R Y3 SO, R Y3 SO2, 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)Independently selected from the group consisting of SO2; In formula ULM-a, T is an optionally substituted alkyl, -(CH2) n -Base, optionally substituted linear, branched-(CH2) n -O-C1-C6 alkyl, or optionally substituted -(CH2) n Selected from the group of -O-heterocyclyls, where each methylene group is a halogen, methyl, optionally substituted alkoxy, a linear or branched C1-C6 alkyl group optionally substituted with one or more halogens, or C(O)NR 1 R 1a , or NR 1 R 1a It is optionally substituted with one or two substituents selected from the group, or R 1 and R 1a These combine to form a heterocycle with any substitutions, or a side chain of an -OH group or an optionally substituted amino acid; W of formula ULM-a 4 This is an arbitrarily substituted -NR 1 -T-aryl (wherein the formula, the aryl group may be optionally substituted with an optionally substituted 5- to 6-membered heteroaryl or optionally substituted with an optionally substituted aryl), optionally substituted -NR1-T-heteroaryl group having an optionally substituted aryl or optionally substituted heteroaryl, or optionally substituted -NR1-T-heterocyclic ring (wherein the formula, -NR1 is X 2 It is covalently bonded to R1, where R1 is H or CH3, preferably H; n is 0 to 6, often 0, 1, 2, or 3, preferably 0 or 1; [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.) Includes a chemical structure selected from.

[0077] In any aspect or embodiment described herein, T is optionally substituted alkyl, -(CH2) n - Selected from the group consisting of groups, where each methylene group is a halogen, methyl, optionally substituted alkoxy, or a linear or branched C1-C6 alkyl group optionally substituted with one or more halogens, C(O)NR 1 R 1a , or NR 1 R 1a It is optionally substituted with one or two substituents selected from the group, or R 1 and R 1a These combine to form an optionally substituted heterocycle, or an -OH group or an optionally substituted amino acid side chain; n is 0 to 6, often 0, 1, 2, or 3, preferably 0 or 1.

[0078] In any aspect or embodiment described herein, the W of formula ULM-a 4 teeth, [ka] (In the formula, W 5 is arbitrarily substituted (for example, W 5 (wherein is an optionally substituted phenyl, optionally substituted naphthyl, or optionally substituted 5-10 member heteroaryl) (for example, W 5 (Optionally substituted with one or more [e.g., 1, 2, 3, 4, or 5] halo, CN, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkoxy, hydroxy, or optionally substituted haloalkoxy), R 14a、 R 14b H, haloalkyl (e.g., fluoroalkyl), optionally substituted alkyl, optionally substituted alkoxy, optionally substituted hydroxylalkyl, optionally substituted alkylamine, optionally substituted heteroalkyl, optionally substituted alkyl-heterocycloalkyl, optionally substituted alkoxy-heterocycloalkyl, COR 26 CONR 27a R27b NHCOR 26 , or NHCH3COR 26 Each of the groups is independently selected; R 14a and R 14b The other is H; or R 14a、 R 14b These, together with the carbon atoms to which they are bonded, form optionally substituted 3- to 5-membered cycloalkyl, heterocycloalkyl, spirocycloalkyl, or spiroheterocyclyl, where the spiroheterocyclyl is not an epoxide or aziridine; o is an integer between 0 and 4 (for example, 0, 1, 2, 3, or 4); R 16 (This is independently selected from the group consisting of halo, optionally substituted alkyl, optionally substituted haloalkyl, hydroxy, or optionally substituted haloalkoxy.) That is the case.

[0079] In any aspect or embodiment described herein, the W of formula ULM-a 5 This is selected from the group of optionally substituted phenyl, optionally substituted naphthyl, or optionally substituted 5-10 member heteroaryls (e.g., W 5 (Optionally substituted with one or more [e.g., 1, 2, 3, 4, or 5] halo, CN, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkoxy, hydroxy, or optionally substituted haloalkoxy), R of formula ULM-a 15 H, halogen, CN, OH, NO2, NR 14a R 14b , OR 14a CONR 14a R 14b , NR 14a COR 14b SO2NR 14a R 14b , NR 14a SO2R 14bSelected from the group consisting of optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy; optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, or optionally substituted cycloheteralkyl.

[0080] In any aspect or embodiment described herein, W for use in this disclosure 4 The substituents also include W, which is found in the specific compounds disclosed herein, in particular (but not limited to the specific compounds disclosed herein). 4 Substituents are included. These W 4 Each of the substituents may also be any number of W as disclosed herein. 3 It can be used in conjunction with substituents.

[0081] In any aspect or embodiment described herein, ULM-a has 0 to 3 R in the pyrrolidine moiety. P It is arbitrarily substituted by each R. P These are independently H, halo, -OH, C1-3 alkyl, and C=O.

[0082] In any aspect or embodiment described herein, the W of formula ULM-a 3 , W 4 It can be independently covalently bonded to a linker bonded to one or more PTM groups. [ka] This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.

[0083] In any aspect or embodiment described herein, ULM is VHL, and structure: [ka] (In the formula, W of formula ULM-b 3is an optionally substituted aryl, an optionally substituted heteroaryl, or [ka] Selected from the group; R9 and R of formula ULM-b 10 independently of R9, 10 , and the carbon atoms to which they are bonded form optionally substituted cycloalkyl groups; R of formula ULM-b 11 This includes arbitrarily substituted heterocyclyls, arbitrarily substituted alkoxys, arbitrarily substituted heteroaryls, and arbitrarily substituted aryls. [ka] Selected from the group; R of formula ULM-b 12 is selected from the group consisting of H or optionally substituted alkyl groups; R of formula ULM-b 13 is selected from the group consisting of 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 of formula ULM-b 14a、 R 14b H, haloalkyl (e.g., fluoroalkyl), optionally substituted alkyl, optionally substituted alkoxy, aminomethyl, alkylaminomethyl, alkoxymethyl, optionally substituted hydroxylalkyl, optionally substituted alkylamine, optionally substituted heteroalkyl, optionally substituted alkyl-heterocycloalkyl, optionally substituted alkoxy-heterocycloalkyl, CONR 27a R 27b CH2NHCOR 26, or (CH2)N(CH3)COR 26 Each of the groups is independently selected; R 14a and R 14b The other is H; or R 14a、 R 14b These, together with the carbon atoms to which they are bonded, form optionally substituted 3- to 6-membered cycloalkyl, heterocycloalkyl, spirocycloalkyl, or spiroheterocyclyl, where the spiroheterocyclyl is not an epoxide or aziridine; W of formula ULM-b 5 This is selected from the group of optionally substituted phenyl or optionally substituted 5-10 member heteroaryls (e.g., W 5 (Optionally substituted with one or more [e.g., 1, 2, 3, 4, or 5] halo, CN, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkoxy, hydroxy, or optionally substituted haloalkoxy), R of formula ULM-b 15 H, halogen, CN, OH, NO2, NR 14a R 14b , OR 14a CONR 14a R 14b , NR 14a COR 14b SO2NR 14a R 14b , NR 14a SO2R 14b , selected from the group of optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, or optionally substituted cycloheteralkyl; Each R in formula ULM-b 16 is independently selected from the group consisting of H, CN, halo, optionally substituted alkyl, optionally substituted haloalkyl, hydroxy, or optionally substituted haloalkoxy; In formula ULM-b, o is 0, 1, 2, 3, or 4; R of formula ULM-b 18is independently selected from the group consisting of H, halo, optionally substituted alkoxy, cyano, optionally substituted alkyl, haloalkyl, haloalkoxy, or linker; In formula ULM-b, p is 0, 1, 2, 3, or 4. [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.) It is represented by [this].

[0084] In any aspect or embodiment described herein, the R of formula ULM-b 15 H, halogen, CN, OH, NO2, NR 27a R 27b , OR 27a CONR 27a R 27b , NR 27a COR 27b SO2NR 27a R 27b , NR 27a SO2R 27b , selected from the group of optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl, each R 26 is H, optionally substituted alkyl or NR 27a R 27b Selected independently from; each R 27a and R 27b H is independently a substituted alkyl group, or R 27a and R 27b These, along with the nitrogen atom to which they are bonded, form a 4- to 6-membered heterocycline.

[0085] In any aspect or embodiment described herein, the R of formula ULM-b 15 teeth, [ka] (In the formula, R 17 H, halo, and C as arbitrarily substituted. 3-6 Cycloalkyl, optionally substituted C 1-6 Alkyl, optionally substituted C 1-6 Alkenyl and C 1-6 It is a haloalkyl group; Xa is either S or O. That is the case.

[0086] In any aspect or embodiment described herein, the R of formula ULM-b 17 The compound is selected from the group consisting of methyl, ethyl, isopropyl, and cyclopropyl.

[0087] In any aspect or embodiment described herein, the R of formula ULM-b 15 teeth, [ka] It is selected from the group consisting of the following.

[0088] In any aspect or embodiment described herein, the R of formula ULM-b 11 teeth, [ka] It is selected from the group consisting of the following.

[0089] In any aspect or embodiment described herein, the R of formula ULM-b 14a、 R 14b H, optionally substituted haloalkyl, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted hydroxylalkyl, optionally substituted alkylamine, optionally substituted heteroalkyl, optionally substituted alkyl-heterocycloalkyl, optionally substituted alkoxy-heterocycloalkyl, CH2OR 30 CH2NHR 30 CH2NCH3R 30 CONR 27a R 27b CH2CONR27a R 27b CH2NHCOR 26 , or CH2NCH3COR 26 Each of the groups is independently selected; R 14a and R 14b The other is H; or R 14a、 R 14b These, together with the carbon atoms to which they are bonded, form optionally substituted 3-6 membered cycloalkyl, heterocycloalkyl, spirocycloalkyl, or spiroheterocyclyl, and the spiroheterocyclyl is not an epoxide or aziridine, and the spirocycloalkyl or spiroheterocycloalkyl itself is alkyl, haloalkyl, or -COR 33 (In the formula, R 33 (which is optionally substituted with alkyl or haloalkyl) R 30 R is selected from H, alkyl, alkynylalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, arylalkyl or heteroarylalkyl (further optionally substituted); R 26 and R 27 This is as stated above.

[0090] In any aspect or embodiment described herein, the R of formula ULM-b 15 H, halogen, CN, OH, NO2, NR 27a R 27b , OR 27a CONR 27a R 27b , NR 27a COR 27b SO2NR 27a R 27b , NR 27a SO2R 27b, optionally substituted alkyl, optionally substituted haloalkyl (e.g., optionally substituted fluoroalkyl), optionally substituted haloalkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl, wherein any substitution of the aryl, heteroaryl, cycloalkyl and heterocycloalkyl is CH2OR 30 CH2NHR 30 CH2NCH3R 30 CONR 27a R 27b CH2CONR 27a R 27b CH2NHCOR 26 CH2NCH3COR 26 or [ka] R 26 , R 27 , R 30 and R 14 a is as described above.

[0091] In any aspect or embodiment described herein, the R of formula ULM-b 14a、 R 14b H, optionally substituted haloalkyl, optionally substituted alkyl, CH2OR 30 CH2NHR 30 CH2NCH3R 30 CONR 27a R 27b CH2CONR 27a R 27b CH2NHCOR 26 , or CH2NCH3COR 26 Each of the groups is independently selected; R 14a and R 14b The other is H; or R 14a、 R 14bThese, together with the carbon atoms to which they are bonded, form optionally substituted 3- to 6-membered spirocycloalkyl or spiroheterocyclyls, and the spiroheterocyclyl is not an epoxide or aziridine, and the spirocycloalkyl or spiroheterocycloalkyl itself is alkyl, haloalkyl, or -COR 33 It is arbitrarily replaced in R 33 is alkyl or haloalkyl, and R 30 is selected from H, alkyl, alkynylalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, arylalkyl or heteroarylalkyl (further optionally substituted); R of formula ULM-b 15 H, halogen, CN, OH, NO2, NR 27a R 27b , OR 27a CONR 27a R 27b , NR 27a COR 27b SO2NR 27a R 27b , NR 27a SO2R 27b , selected from optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl, wherein any substitution of the aryl, heteroaryl, cycloalkyl and heterocycloalkyl is CH2OR 30 CH2NHR 30 CH2NCH3R 30 CONR 27a R 27b CH2CONR 27a R 27b CH2NHCOR 26 CH2NCH3COR 26 or [ka] R 26 , R 27 , R 30and R 14 a is as described above.

[0092] In any aspect or embodiment described herein, ULM is [ka] (In the formula, In formulas ULM-c, ULM-d, and ULM-e, R1 is H, ethyl, isopropyl, tert-butyl, sec-butyl, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl; optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted hydroxyalkyl, optionally substituted heteroaryl, or haloalkyl; Formulas ULM-c, ULM-d, and ULM-e 14a is H, haloalkyl, optionally substituted alkyl, methyl, fluoromethyl, hydroxymethyl, ethyl, isopropyl, or cyclopropyl; Formulas ULM-c, ULM-d, and ULM-e 15 This is selected from the group consisting of H, halogens, CN, OH, NO2, optionally substituted heteroaryls, optionally substituted aryls; optionally substituted alkyls, optionally substituted haloalkyls, optionally substituted haloalkoxys, optionally substituted cycloalkyls, or optionally substituted cycloheteralkyls; In formulas ULM-c, ULM-d, and ULM-e, X is C, CH2, or C=O. In formulas ULM-c, ULM-d, and ULM-e, R3 is a 5- or 6-membered heteroaryl that is absent or optionally substituted; [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.) It has a chemical structure selected from the group.

[0093] In any aspect or embodiment described herein, ULM has a chemical structure: [ka] (In the formula, R of formula ULM-f 14a is H, haloalkyl, optionally substituted alkyl, methyl, fluoromethyl, hydroxymethyl, ethyl, isopropyl, or cyclopropyl; In formula ULM-f, R9 is H, R of formula ULM-f 10 These are H, ethyl, isopropyl, tert-butyl, sec-butyl, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl; R of formula ULM-f 11 teeth, [ka] ; or an optionally substituted heteroaryl; In formula ULM-f, p is 0, 1, 2, 3, or 4; Each R in formula ULM-f 18 These are independently halo, optionally substituted alkoxy, cyano, optionally substituted alkyl, haloalkyl, haloalkoxy, or linker; R of formula ULM-f 12 H is C=O; R of formula ULM-f 13 is H, an optionally substituted alkyl, an optionally substituted alkylcarbonyl, an optionally substituted (cycloalkyl)alkylcarbonyl, an optionally substituted aralkylcarbonyl, an optionally substituted arylcarbonyl, an optionally substituted (heterocyclyl)carbonyl, or an optionally substituted aralkyl. R of formula ULM-f 15 This is selected from the group consisting of H, halogens, Cl, CN, OH, NO2, optionally substituted haloalkyls, optionally substituted heteroaryls, and optionally substituted aryls; [ka] Selected from the group consisting of; Formula ULM-f [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.) It has a base due to [the following].

[0094] In any aspect or embodiment described herein, VLM is an R group (for example, R P , R 1 , R 1a , R 1b , R Y3 , R Y4 , R9, R 10 , R 11 , R 12 , R 13 , R 14a , R 14b , R 15 , R 16 , R 17 , R 18 , R 26 , R27a, R 27b , R 30 , R 33 ), W 3 , W 4 , W 5 , X, X 1 , X 2 , X 3 , or covalently bonded to a PTM or chemical linker group (L) via T.

[0095] In any aspect or embodiment described herein, VLM is R P , R 1 , R 1a , R 1b , R Y3 , R Y4 , R9, R 10 , R 11 , R 12 , R 13 , R 14a , R 14b , R 15 , R16 , R 17 , R 18 , R 26 , R27a, R 27b , R 30 , R 33 , W 3 , W 4 , W 5 , X, X 1 , X 2 , X 3 , or covalently bonded to a PTM or chemical linker group (L) via T.

[0096] In any aspect or embodiment described herein, R P , R 1 , R 1a , R 1b , R Y3 , R Y4 , R9, R 10 , R 11 , R 12 , R 13 , R 14a , R 14b , R 15 , R 16 , R 17 , R 18 , R 26 , R27a, R 27b , R 30 , R 33 , W 3 , W 4 , X, X 1 , X 2 , X 3 , or T may independently be covalently bonded to a linker and / or a linker bonded to one or more PTM, ULM, and VLM groups.

[0097] In any aspect or embodiment described herein, the ULM has the following structure: [ka] [ka] (In the formula, [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.) Selected from.

[0098] In any aspect or embodiment described herein, the ULM has the following structure: [ka] (In the formula, n is either 0 or 1, [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.) Selected from.

[0099] In any aspect or embodiment described herein, the ULM has the following structure: [ka] [ka] [ka] (In the formula, the phenyl rings in ULM-a1 to ULM-a15, ULM-b1 to ULM-b12, ULM-c1 to ULM-c15 and ULM-d1 to ULM-d9 are optionally substituted with fluorine, lower alkyl, and alkoxy groups.) [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to ULM-a.) Selected from.

[0100] In any aspect or embodiment described herein, the phenyl rings in ULM-a1 to ULM-a15, ULM-b1 to ULM-b12, ULM-c1 to ULM-c15 and ULM-d1 to ULM-d9 may be functionalized as esters to form part of a prodrug.

[0101] In any embodiment or configuration described herein, the hydroxyl groups on the pyrrolidine rings of ULM-a1 to ULM-a15, ULM-b1 to ULM-b12, ULM-c1 to ULM-c15, and ULM-d1 to ULM-d9 each contain an ester-linked prodrug moiety.

[0102] In any aspect or embodiment described herein, ULM and ULM', if present, each independently have the following chemical structure: [ka] or a pharmaceutically acceptable salt thereof (in the formula, R1 is H, an optionally substituted alkyl, or an optionally substituted cycloalkyl; R3 is an optionally substituted 5-6 member heteroaryl; W 5 is optionally substituted phenyl, optionally substituted naphthyl, or optionally substituted pyridinyl; R 14a and R 14b One of them is H, optionally substituted alkyl, optionally substituted haloalkyl (e.g., fluoroalkyl), optionally substituted alkoxy, optionally substituted hydroxylalkyl, optionally substituted alkylamine, optionally substituted heteroalkyl, optionally substituted alkyl-heterocycloalkyl, optionally substituted alkoxy-heterocycloalkyl, COR 26 CONR 27a R 27b NHCOR 26 , or NHCH3COR 26 And; R 14a and R14b The other is H; or R 14a , R 14b These, together with the carbon atoms to which they are bonded, form optionally substituted 3- to 6-membered cycloalkyl, heterocycloalkyl, spirocycloalkyl, or spiroheterocyclyl, where the spiroheterocyclyl is not an epoxide or aziridine; R 15 CN, optionally substituted fluoroalkyl, [ka] Arbitrarily replaced [ka] (for example, [ka] (In the formula, R 28a (is a halo, optionally substituted alkyl or fluoroalkyl), or [ka] and; Each R 16 This is independently selected from halo, CN, optionally substituted alkyl, optionally substituted haloalkyl, hydroxy, or haloalkoxy; Each R 26 These are independently H, optionally substituted alkyl or NR 27a R 27b and; Each R 27a and R 27b H is independently a substituted alkyl group, or R 27a and R 27b They, together with the nitrogen atom to which they are bonded, form a 4-6 membered heterocycline; R 28is H, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted heteroalkyl, optionally substituted alkylamine, optionally substituted hydroxyalkyl, amine, optionally substituted alkynyl, or optionally substituted cycloalkyl; o is 0, 1, or 2; [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.) It is a basis of [the system].

[0103] In any aspect or embodiment described herein, ULM is defined by the formula: [ka] (In the formula, X 4 , X 5 , and X 6 Each of them is selected from CH and N, with two or fewer being N; R 1 It is a C1-6 alkyl group; R3 is an optionally substituted 5-6 member heteroaryl; R 14a and R 14b One of them is H, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkoxy, optionally substituted hydroxylalkyl, optionally substituted alkylamine, optionally substituted heteroalkyl, optionally substituted alkyl-heterocycloalkyl, optionally substituted alkoxy-heterocycloalkyl, COR 26 CONR 27a R 27b NHCOR 26 , or NHCH3COR 26 And; R 14a and R 14b The other is H; or R 14a and R 14bThese, together with the carbon atoms to which they are bonded, form optionally substituted 3- to 5-membered cycloalkyl, heterocycloalkyl, spirocycloalkyl, or spiroheterocyclyl, where the spiroheterocyclyl is not an epoxide or aziridine; Each R 27a and R 27b H or C 1-6 It is alkyl; q is 1, 2, 3, or 4; R 15 teeth, [ka] or CN; R 28 H, methyl, CH2N(Me)2, CH2OH, CH2O(C 1-4 Alkyl), CH2NHC(O)C 1-4 Alkyl, NH2, [ka] and; R 28C is H, methyl, fluoro, or chloro; R 16 H, C 1-4 Alkyl, fluoro, chloro, CN, or C 1-4 It is an alkoxy; [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.) It belongs to them.

[0104] In any aspect or embodiment described herein, R 14a and R 14b H, C 1-4 Alkyl, C 1-4 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Hydroxyalkyl, C1-4 Alkyloxyalkyl, C 1-4 Alkyl-NR 27a R 27b and CONR 27a R 27b Selected from.

[0105] In any aspect or embodiment described herein, R 14a and R 14b At least one of them is H (for example, R 14a and R 14b (Both are H).

[0106] In any aspect or embodiment described herein, R 14a and R 14b At least one of them is an optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkoxy, optionally substituted hydroxylalkyl, optionally substituted alkylamine, optionally substituted heteroalkyl, optionally substituted alkyl-heterocycloalkyl, optionally substituted alkoxy-heterocycloalkyl, COR 26 CONR 27a R 27b NHCOR 26 , or NHCH3COR 26 Alternatively, in any aspect or embodiment described herein, R 14a and R 14b One of them is an optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkoxy, optionally substituted hydroxylalkyl, optionally substituted alkylamine, optionally substituted heteroalkyl, optionally substituted alkyl-heterocycloalkyl, optionally substituted alkoxy-heterocycloalkyl, COR 26 CONR 27a R 27b NHCOR 26 , or NHCH3COR 26 And; R 14a and R 14b The other is H.

[0107] In any aspect or embodiment described herein, R 14a and R 14b Together with the carbon atoms to which they are bonded, [ka] (In the formula, R 23 H, C 1-4 Alkyl, -C(O)C 1-4 Forms an alkyl group (selected from alkyl groups).

[0108] In any aspect or embodiment described herein, ULM and, if present, ULM' are each independently defined as having the following chemical structure: [ka] or a pharmaceutically acceptable salt thereof (in the formula, X is either CH or N; R1 is H, an optionally substituted alkyl, or an optionally substituted cycloalkyl; R3 is an optionally substituted 5-6 member heteroaryl; R 14a and R 14b One of them is H, optionally substituted alkyl, optionally substituted haloalkyl (e.g., fluoroalkyl), optionally substituted alkoxy, optionally substituted hydroxylalkyl, optionally substituted alkylamine, optionally substituted heteroalkyl, optionally substituted alkyl-heterocycloalkyl, optionally substituted alkoxy-heterocycloalkyl, COR 26 CONR 27a R 27b NHCOR 26 , or NHCH3COR 26 And; R 14a and R 14b The other is H; or R 14a、 R 14bThese, together with the carbon atoms to which they are bonded, form optionally substituted 3- to 6-membered cycloalkyl, heterocycloalkyl, spirocycloalkyl, or spiroheterocyclyl, where the spiroheterocyclyl is not an epoxide or aziridine; R 15 CN, optionally substituted fluoroalkyl, [ka] Arbitrarily replaced [ka] (for example, [ka] (In the formula, R 28a (is a halo, optionally substituted alkyl or fluoroalkyl), or [ka] and; Each R 26 These are independently H, optionally substituted alkyl or NR 27a R 27b and; Each R 27a and R 27b H is independently a substituted alkyl group, or R 27a and R 27b They, together with the nitrogen atom to which they are bonded, form a 4-6 membered heterocycline; R 28 is H, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted heteroalkyl, optionally substituted alkylamine, optionally substituted hydroxyalkyl, amine, optionally substituted alkynyl, or optionally substituted cycloalkyl; [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.) It is a basis of [the system].

[0109] In any aspect or embodiment described herein, R1 is C 1-6 It is alkyl.

[0110] In any of the embodiments or aspects described herein, R 14a and R 14b One of them is H, C 1-6 Alkyl, C 1-6 Haloalkyl, optionally substituted C 1-4 Alkylamine, C 1-6 Alkoxy, (CH2) q C 1-6 Alkoxy, (CH2) q C 1-6 Alkoxy-C3-C7 heterocycloalkyl, (CH2) q OH, (CH2) q NR 27a R 27b , (CH2) q NHCOC 1-6 Alkyl, C 3-6 Cycloalkyl, or NR 27a R 27b and; each R 26 H and C are independent of each other. 1-6 Alkyl or NR 27a R 27b and; each R 27a and R 27b H or C 1-6 It is alkyl; q is 1, 2, 3, or 4.

[0111] In any of the embodiments or aspects described herein, R 14a and R 14b One of them is H, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 alkoxy, optionally substituted C 1-4 Alkylamine, (CH2)q C 1-6 Alkoxy, (CH2) q C 1-6 Alkoxy-C3-C7 heterocycloalkyl, (CH2) q OH, (CH2) q NR 27a R 27b , (CH2) q NHCOC 1-6 Alkyl, C 3-6 Cycloalkyl, or NR 27a R 27b and; each R 26 H and C are independent of each other. 1-4 Alkyl or NR 27a R 27b and; each R 27a and R 27b H or C 1-4 It is alkyl; q is 1 or 2.

[0112] In any of the embodiments or aspects described herein, R 28 C 1-6 Alkyl, C 3-6 Cycloalkyl, C 1-6 Haloalkyl, (CH2) q OC 1-6 Alkyl, (CH2) q OH, (CH2) q NR 27a R 27b , (CH2) q NHCOC 1-6 alkyl, or [ka] and; R 29 H, C 1-6 Alkyl, NR 27a R 27b or q NHCOC 1-6 It is alkyl; q is either 1 or 2.

[0113] In any of the embodiments or aspects described herein, R 3is isoxazolyl, 4-chloroisoxazolyl, 4-fluoroisoxazolyl, or pyrazolyl. In any of the embodiments or models described herein, X is CH.

[0114] In any aspect or embodiment described herein, ULM is defined by the formula: [ka] or a pharmaceutically acceptable salt thereof (In the formula, R1, R 14a and R 14b This is as described herein; X is either CH or N; R 30 is H, F, or Cl; R 16 H, C 1-4 Alkyl, fluoro, chloro, CN, or C 1-4 It is an alkoxy; R 28 H, methyl, CH2N(Me)2, CH2OH, CH2O(C 1-4 Alkyl), CH2NHC(O)C 1-4 Alkyl, NH2, [ka] and; [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.) Follow the rules.

[0115] In any aspect or embodiment described herein, ULM is defined by the formula: [ka] or a pharmaceutically acceptable salt thereof (In the formula, R1, R 14a , R 14b Each of these is as described herein; R 30 is H, F, or Cl; [ka] (This indicates a binding site for a chemical linker that binds at least one PTM, another ULM (ULM'), or at least one PTM or ULM' or both to the ULM.) Follow the rules.

[0116] In any aspect or embodiment described herein, VLM is an R group (for example, R 1 , R 3 , R 14a , R 14b , R 15 , R 16 , R 23 , R 26 , R 27a , R 27b , R 28 , R 28a , R 28C , R 29 , R 30 ), X, X 4 , X 5 , or X 6 It is covalently bonded to the PTM or chemical linker group (L) via this.

[0117] In any aspect or embodiment described herein, VLM is R 1 , R 3 , R 14a , R 14b , R 15 , R 16 , R 23 , R 26 , R 27a , R 27b , R 28 , R 28a , R 28C , R 29 , R 30 , X, X 4 , X 5 , or X 6It is covalently bonded to the PTM or chemical linker group (L) via this.

[0118] In any aspect or embodiment described herein, R 1 , R 3 , R 14a , R 14b , R 15 , R 16 , R 23 , R 26 , R 27a , R 27b , R 28 , R 28a , R 28C , R 29 , R 30 , X, X 4 , X 5 , or X 6 It can independently be covalently bonded to a linker and / or a linker bonded to one or more PTM, ULM, and VLM groups.

[0119] In any of the embodiments or models described herein, the ULM (or ULM', if present) described herein may be a pharmaceutically acceptable salt, enantiomer, diastereomer, solvate, or polymorph thereof. Also in any of the embodiments or models described herein, the ULM (or ULM', if present) may be bonded to the PTM directly via bonding or by a chemical linker.

[0120] Exemplary linker In any embodiment or configuration described herein, the compounds described herein include PTM chemically linked to ULM (e.g., VLM) via a chemical linker (L). In a given embodiment, the linker group L is one or more covalently bonded structural units (e.g., -A L 1… (A L ) q -or-(A L ) q -)(In the formula, A L 1 is a group bonded to PTM, (A L ) qIt includes a group that is bound to ULM.

[0121] In any aspect or embodiment described herein, the linker(L) connection to the ULM (e.g., VLM) is a stable L-ULM connection. For example, in any aspect or embodiment described herein, if the linker(L) and ULM are connected via a heteroatom (e.g., N, O, S), any additional heteroatom, if present, is separated, for example, by at least a carbon atom (e.g., -CH2-), such as an acetal or aminal group. As a further example, in any aspect or embodiment described herein, if the linker(L) and ULM are connected via a heteroatom, the heteroatom is not part of the ester.

[0122] In any aspect or embodiment described herein, the linker group L is bonded or of formula -(A L ) q - is a chemical linker group represented by -, where A is the chemical site and q is an integer from 1 to 100 (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, 5 L is covalently bonded to both PTM and ULM, providing binding of PTM to the protein target and binding of ULM to the E3 ubiquitin ligase to achieve ubiquitination of the target protein. (0, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or 80)

[0123] In any aspect or embodiment described herein, the linker group L is bonded or of formula -(A L ) q- is a chemical linker group represented by -, where A is a chemical site, q is an integer from 6 to 30 (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, or 25), and L is covalently bonded to both PTM and ULM, providing binding of PTM to a protein target sufficiently close to result in ubiquitination of the target protein and binding of ULM to the E3 ubiquitin ligase.

[0124] In any aspect or embodiment described herein, the linker group L is -(A L ) q -(In the formula, (A L ) q This is the base that connects ULM (e.g., VLM) to PTM (KTM); The linker's q is an integer greater than or equal to 1; Each A L , combine, CR L1 R L2 O, S, SO, SO2, NR L3 SO2NR L3 , SONR L3 CONR L3 , NR L3 CONR L4 , NR L3 SO2NR 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(=CNO2)NR L4 , 1 to 6 R L1 and / or R L2 C arbitrarily substituted in the base 3-11 Cycloalkyl, 1 to 9 R L1 and / or R L2 C arbitrarily substituted in the base 5-13Spirocycloalkyl, 1 to 6 R L1 and / or R L2 C arbitrarily substituted in the base 3-11 Heterocycline, 1-8 R L1 and / or R L2 C arbitrarily substituted in the base 5-13 Spiroheterocyclyl, 1-6 R L1 and / or R L2 Aryl compounds arbitrarily substituted with a base, 1 to 6 R L1 and / or R L2 Independently selected from the group consisting of heteroaryls arbitrarily substituted with R L1 or R L2 Each of these can be independently and arbitrarily connected to other groups, forming 1 to 4 R L5 Forms optionally substituted cycloalkyl and / or heterocyclyl moieties with a group; R L1 , R L2 , R L3 , R L4 and R L5 These are H, Halo, and C, respectively, independently. 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 Heterocycline, OC 3-8 Cycloalkyl, SC 3-8 Cycloalkyl, NHC 3-8 Cycloalkyl, N(C 3-8 Cycloalkyl)2, N(C 3-8 Cycloalkyl)(C 1-8 Alkyl), OH, NH2, SH, SO2C 1-8 alkyl, P(O)(OC 1-8 Alkyl)(C 1-8 alkyl), P(O)(OC 1-8 Alkyl)2, CC-C 1-8 Alkyl, CCH, CH=CH(C 1-8 Alkyl), C(C 1-8 Alkyl)=CH(C 1-8Alkyl), 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, CO2H, Halogen, CN, CF3, CHF2, CH2F, NO2, SF5, SO2NHC 1-8 Alkyl, SO2N(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, NHCONH2, N(C 1-8 Alkyl)SO2NH(C 1-8 Alkyl), N(C 1-8 Alkyl)SO2N(C 1-8 Alkyl)2, NHSO2NH(C 1-8 Alkyl), NHSO2N(C 1-8 Alkyl)2, NHSO2NH2) That is the case.

[0125] In any aspect or embodiment described herein, q is an integer of 1 or more.

[0126] In any aspect or embodiment described herein, for example, if the linker q is greater than 2, (A L ) q is, A L 1 and (A L ) q The linker is a base that binds PTM to ULM.

[0127] In any aspect or embodiment described herein, for example, when the linker q is 2, A L2 is A L It is a base connected to 1 and ULM.

[0128] In any aspect or embodiment described herein, for example, when the linker q is 1, the structure of the linker group L is -A L 1- and A L 1 is a base that connects the ULM site to the PTM site.

[0129] In any aspect or embodiment described herein, the unit A of the linker (L) L teeth, -NR(CH2) n -(lower alkyl)-, -NR(CH2) n -(lower alkoxy)-, -NR(CH2) n -(lower alkoxyl)-OCH2-, -NR(CH2) n -(lower alkoxyl)-(lower alkyl)-OCH2-,-NR(CH2) n -(cycloalkyl)-(lower alkyl)-OCH2-,-NR(CH2) n -(heterocycloalkyl)-, -NR(CH2CH2O) n -(lower alkyl)-O-CH2-, -NR(CH2CH2O) n -(heterocycloalkyl)-O-CH2-, -NR(CH2CH2O) n -aryl-O-CH2-, -NR(CH2CH2O) n -(heteroaryl)-O-CH2-, -NR(CH2CH2O) n -(cycloalkyl)-O-(heteroaryl)-O-CH2-,-NR(CH2CH2O) n -(cycloalkyl)-O-aryl-O-CH2-,-NR(CH2CH2O) n -(lower alkyl)-NH-aryl-O-CH2-,-NR(CH2CH2O) n -(lower alkyl)-O-aryl-CH2,-NR(CH2CH2O) n -Cycloalkyl-O-aryl-,-NR(CH2CH2O) n -Cycloalkyl-O-(heteroaryl)l-,-NR(CH2CH2)n -(cycloalkyl)-O-(heterocyclyl)-CH 2、 -NR(CH2CH2) n -(heterocyrill)-(heterocyrill)-CH2, and -N(R1R2)-(heterocyrill)-CH2 (wherein, Linker's n can be between 0 and 10; The R in the linker can be H or a lower alkyl group; Linker R1 and R2 may form a ring with connectivity N. It includes a group represented by a general structure selected from the group consisting of the following.

[0130] In any aspect or embodiment described herein, the linker (L) is optionally replaced with C1-C 50 Alkyl (for example, C1, C2, C3, C4, C5, C6, C7, C8, C9, 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 , C 24 , C 25 , C 26 , C 27 , C 28 , C 29 , C 30 , C 31 , C 32 , C 33 , C 34 , C 35 , C 36 , C 37 , C 38 , C 39 , C 40 , C 41 , C 42 , C 43 , C 44 , C 45 , C 46 , C 47 , C 48 , C49 , or C 50 The alkyl group includes all implied secondary ranges, e.g., C1-C10, C1-C20; C2-C10, C2-20; C10-C20, C10-C50, etc.), and each carbon is optionally, independently, substituted or replaced with (1) a heteroatom selected from N, O, S, P, or Si atoms having an appropriate number of hydrogens, substitutions, or both, to align the valencies; (2) optionally substituted cycloalkyl or bicyclic cycloalkyl; (3) optionally substituted heterocycloalkyl or bicyclic heterocycloalkyl; (4) optionally substituted aryl or bicyclic aryl; or (5) optionally substituted heteroaryl or bicyclic heteroaryl. In any aspect or embodiment described herein, the linker (L) does not have heteroatom-heteroatom bonds (e.g., heteroatoms are not covalently bonded or located adjacent to each other).

[0131] In any aspect or embodiment described herein, the linker (L) is optionally replaced with C1-C 50 Alkyl (for example, C1, C2, C3, C4, C5, C6, C7, C8, C9, 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 , C 24 , C 25 , C 26 , C 27 , C 28 , C 29 , C 30 , C 31 , C 32 , C 33 , C 34 , C 35 , C 36 , C 37 , C 38 , C 39, C 40 , C 41 , C 42 , C 43 , C 44 , C 45 , C 46 , C 47 , C 48 , C 49 , or C 50 alkyl) (wherein, each carbon is CR L1 R L2 , O, S, SO, SO2, NR L3 , SO2NR L3 , SONR L3 , CONR L3 , NR L3 CONR L4 , NR L3 SO2NR 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(=CNO2)NR L4 , 1 to 6 R L1 and / or R L2 groups optionally substituted C 3-11 cycloalkyl, 1 to 9 R L1 and / or R L2 groups optionally substituted C 5-13 spirocycloalkyl, 1 to 6 R L1 and / or R L2 groups optionally substituted C 3-11 heterocyclyl, 1 to 8 R L1 and / or R L2 groups optionally substituted C 5-13 spiroheterocyclyl, 1 to 6 R L1 and / or R L2 groups optionally substituted aryl, 0 to 6 R L1 and / or R L2The heteroaryl group is arbitrarily substituted or replaced by the base, R L1 or R L2 Each of these can be independently and arbitrarily connected to other groups, forming 1 to 4 R L5 Forms optionally substituted cycloalkyl and / or heterocyclyl moieties with a group; R L1 , R L2 , R L3 , R L4 and R L5 These are H, Halo, and C, respectively, independently. 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 Heterocycline, OC 3-8 Cycloalkyl, SC 3-8 Cycloalkyl, NHC 3-8 Cycloalkyl, N(C 3-8 Cycloalkyl)2, N(C 3-8 Cycloalkyl)(C 1-8 Alkyl), OH, NH2, SH, SO2C 1-8 alkyl, P(O)(OC 1-8 Alkyl)(C 1-8 alkyl), P(O)(OC 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, CO2H, Halogen, CN, CF3, CHF2, CH2F, NO2, SF5, SO2NHC 1-8 Alkyl, SO2N(C 1-8 Alkyl) 2, SONHC 1-8Alkyl, 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, NHCONH2, N(C 1-8 alkyl)SO2NH(C 1-8 alkyl), N(C 1-8 alkyl)SO2N(C 1-8 alkyl)2, NHSO2NH(C 1-8 alkyl), NHSO2N(C 1-8 alkyl)2, NHSO2NH2) is included.

[0132] In any aspect or embodiment described herein, the linker group is optionally substituted C1-C 50 alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, 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 , C 24 , C 25 , C 26 , C 27 , C 28 , C 29 , C 30 , C 31 , C 32 , C 33 , C 34 , C 35 , C 36 , C 37 , C 38 , C 39 , C40 ,W 41 ,W 42 ,W 43 ,W 44 ,W 45 ,W 46 ,W 47 ,W 48 ,W 49 、またはC 50Alkyl (including all implied secondary ranges, e.g., C1-C10, C1-C20; C2-C10, C2-20; C10-C20, C10-C50, etc.)), where each carbon atom is an O, N, S, P, or Si atom having an appropriate number of hydrogens, substitutions (e.g., OH, halo, alkyl, methyl, ethyl, haloalkyl, hydroxyalkyl, alkoxy, methoxy, etc.), or both, to balance the valence; optionally substituted aryl (e.g., optionally substituted C5 or C6 aryl) or bicyclic aryl (e.g., optionally substituted C5-C20 bicyclic heteroaryl); optionally substituted heteroaryl (e.g., optionally substituted C5 or C6 heteroaryl) or bicyclic heteroaryl (e.g., selected from O, N, S, P, or Si having an appropriate number of hydrogens, substitutions (e.g., OH, halo, alkyl, methyl, ethyl, haloalkyl, hydroxyalkyl, alkoxy, methoxy, etc.), or both, to balance the valence; optionally substituted aryl (e.g., optionally substituted C5 or C6 heteroaryl) or bicyclic heteroaryl (e.g., selected from O, N, S, P, or Si having an appropriate number of hydrogens, substitutions (e.g., OH, halo, alkyl, methyl, ethyl, haloalkyl, hydroxyalkyl, alkoxy, methoxy, etc.), or both, to balance the valence) Optionally substituted heteroaryl or bicyclic heteroaryl having one or more heteroatoms; optionally substituted C1-C6 alkyl; optionally substituted C1-C6 alkenyl; optionally substituted C1-C6 alkynyl; optionally substituted cycloalkyl (e.g., optionally substituted C3-C7 cycloalkyl) or bicyclic cycloalkyl (e.g., optionally substituted C5-C20 bicyclic cycloalkyl); or optionally substituted heterocycloalkyl (e.g., optionally substituted 3, 4, 5, 6, or 7-membered heterocyclic groups) or bicyclic heteroalkyl (e.g., optionally substituted heterocycloalkyl or bicyclic heteroalkyl having one or more heteroatoms selected from N, O, S, P, or Si atoms having an appropriate number of hydrogens, substitutions (e.g., OH, halo, alkyl, methyl, ethyl, haloalkyl, hydroxyalkyl, alkoxy, methoxy, etc.), or both, to match the valence.In any aspect or embodiment described herein, the optionally substituted alkyl linker is optionally substituted with one or more OH groups, halo groups, linear or branched C1-C6 alkyl groups (such as methyl or ethyl groups), linear or branched C1-C6 haloalkyl groups, linear or branched C1-C6 hydroxyalkyl groups, or linear or branched C1-C6 alkoxy groups (e.g., methoxy groups).

[0133] In any aspect or embodiment described herein, the linker (L) does not have heteroatom-heteroatom bonds (for example, the heteroatoms are not covalently bonded or are not located adjacent to each other).

[0134] In any aspect or embodiment described herein, the linker (L) contains optionally substituted 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 units, wherein carbon or oxygen may be substituted or replaced with heteroatoms selected from N, S, P, or Si atoms having an appropriate number of hydrogens to match the valence.

[0135] In any aspect or embodiment described herein, linker(L) has the following chemical structure: [ka] (In the formula, Chemical linkage site [ka] This is a binding site for VLM or PTM; Y L2 is a bond, or an unsubstituted or substituted linear or branched C1-C4 alkyl (e.g., optionally substituted with halogen, C1-3 alkyl, methyl, or ethyl); W L3 These are 3- to 7-membered rings (e.g., 4- to 6-membered cycloalkyl or heterocycloalkyl) or 8- to 12-membered spiro rings, each having 0- to 4 heteroatoms (e.g., 0- to 4 heteroatoms independently selected from N, O, and S) and optionally substituted with halogens or methyl groups; Y L3 is a bond or C1-C35 alkyl (C1, C2, C3, C4, C5, C6, C7, C8, C9, 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 , C 24 , C 25 , C 26 , C 27 , C 28 , C 29 , C 30 , C 31 , C 32 , C 33 , C 34 , or C 35 It is alkyl, and one or more C atoms are O, [ka] Alternatively, each carbon is optionally replaced with NH, each carbon is optionally replaced with halogen, =O, methyl or ethyl, and each nitrogen is optionally replaced with halogen, methyl or ethyl; Y L4 It is a bonded, O, or unsubstituted or substituted linear or branched C1-C6 alkyl, where one or more carbons are optionally replaced with O, NH, or NCH3, and optionally substituted with halogens or methyl groups; W L4 This is a 3-8 member ring (for example, a 4-6 member cycloalkyl or heterocycloalkyl, or [ka] Alternatively, they are 5-8 membered spiro rings, each having 0-4 heteroatoms (e.g., 0-4 heteroatoms independently selected from N, O, and S), and optionally substituted with halogens (e.g., F, Cl, Br) or methyl groups; Y L5 (The C1-C6 alkyl group is bonded, unsubstituted, or substituted, and one or more C atoms are optionally replaced by O, and optionally substituted by a halo (e.g., F, Cl, Br), or methyl.) It is represented by [this].

[0136] In any aspect or embodiment described herein, the unit A of the linker (L) L teeth, [ka] [ka] [ka] [ka] (In the formula, [ka] The structure includes a structure selected from the group consisting of (where is a binding point with PTM or VLM).

[0137] In any aspect or embodiment described herein, the unit A of the linker (L) L teeth, [ka] [ka] [ka] [ka] (In the formula, [ka] (This indicates the junction with PTM or VLM.) Includes a structure selected from the group consisting of the following.

[0138] In any aspect or embodiment described herein, the linker (L) has the following structure: [ka] (In the formula, W L1 and W L2 Each is independent of non-existence, R Q A 4-8 membered ring having 0-4 heteroatoms arbitrarily substituted with each R Q These are independently H, halo, OH, CN, CF3, optionally substituted linear or branched C1-C6 alkyl, optionally substituted linear or branched C1-C6 alkoxy, or two R Q The groups, together with the atoms to which they are bonded, form a 4-8 membered ring system containing 0-4 heteroatoms; Y L1 Each is independently bonded, optionally substituted linear or branched C1-C6 alkyl (where one or more C atoms are O or NR). YL1 A C1-C6 alkoxy is a C1-C6 alkene (where any one or more C atoms are replaced by O), a C1-C6 alkyne (where any one or more C atoms are replaced by O), or a C1-C6 alkoxy is a C1-C6 alkoxy (where any one or more C atoms are replaced by O); R YL1 H, or optionally substituted linear or branched C 1-6 It is alkyl; n is between 0 and 10; [ka] (This indicates a binding site to the PTM or ULM site.) Includes structures selected from.

[0139] In any aspect or embodiment described herein, the linker (L) has the following structure: [ka] (In the formula, W L1 and W L2 These are, independently, non-existent, piperazine, piperidine, and morpholine (R Q (which is arbitrarily substituted by) and each R Q These are independently H, -Cl-, -F-, OH, CN, CF3, optionally substituted linear or branched C1-C6 alkyl (e.g., methyl, ethyl), and optionally substituted linear or branched C1-C6 alkoxy (e.g., methoxy, ethoxy); Y L1 Each is independently bonded, optionally substituted linear or branched C1-C6 alkyl (where one or more C atoms are O or NR). YL1 A C1-C6 alkoxy is a C1-C6 alkene (where any one or more C atoms are replaced by O), a C1-C6 alkyne (where any one or more C atoms are replaced by O), or a C1-C6 alkoxy is a C1-C6 alkoxy (where any one or more C atoms are replaced by O); R YL1 H, or optionally substituted linear or branched C 1-6 Alkyl (e.g., methyl, ethyl); n is between 0 and 10; [ka] (This indicates a binding site to the PTM or ULM site.) Includes structures selected from.

[0140] In any aspect or embodiment described herein, the linker (L) has the following structure: [ka] (In the formula, W L1 and W L2 These are, independently, non-existent, aryl, heteroaryl, cyclic, heterocyclic, and C. 1-6 Alkyl (any one or more C atoms are O or NR) YL1 (It is replaced by C) 1-6 Alkenes (where one or more C atoms are replaced by O), C 1-6 Alkynes (where one or more C atoms are replaced by O), bicyclic, biaryl, biheteroaryl, or biheterocyclic, each of which is R Q It is arbitrarily substituted, and each R Q These are independently H, halo, OH, CN, CF3, hydroxyl, nitro, C≡CH, C 2-6 Alkenil, C 2-6 Alkynyl, optionally substituted linear or branched C1-C6 alkyl, optionally substituted linear or branched C1-C6 alkoxy, optionally substituted OC 1-3 Alkyl (e.g., optionally substituted with one or more -F groups), OH, NH2, NR Y1 R Y2 , CN, or 2 R Q The groups, together with the atoms to which they are bonded, form a 4-8 membered ring system containing 0-4 heteroatoms; Y L1 These are, independently, combined, and NR. YL1 , O, S, NR YL2 , CR YL1 R YL2 , C=O, C=S, SO, SO2, optionally substituted linear or branched C1-C6 alkyl (where one or more C atoms are optionally replaced by O); optionally substituted linear or branched C1-C6 alkoxy; Q L It is arbitrarily bridged, with 0 to 6 R Q A 3-6 membered alicyclic, bicyclic, or aromatic ring having 0-4 heteroatoms optionally substituted with each R QThese are independently H, and optionally substituted linear or branched C 1-6 Alkyl (for example, one or more halos, C 1-6 (Optionally substituted with alkoxyl) or two R Q The groups, together with the atoms to which they are bonded, form a 3-8 membered ring system containing 0-2 heteroatoms; R YL1 , R YL2 Each of these is independently H, OH, and optionally substituted linear or branched C. 1-6 Alkyl (for example, one or more halos, C 1-6 (Optionally substituted with alkoxyl), or R 1 , R 2 Together with the atoms to which they are bonded, they form a 3-8 membered ring system containing 0-2 heteroatoms; n is between 0 and 10; [ka] (This indicates a binding site to the PTM or ULM site.) Includes structures selected from.

[0141] In any aspect or embodiment described herein, the linker (L) has the following structure: [ka] (In the formula, W L1 and W L2 These are, independently, absent, cyclohexane, cyclopentane, piperazine, piperidine, morpholine, and C. 1-6 Alkyl (any one or more C atoms are O or NR) YL1 (It is replaced by C) 1-6 Alkenes (where one or more C atoms are replaced by O), C 1-6 Alkenes (where one or more C atoms are replaced by O), or C 1-6 Alkynes (where one or more C atoms are replaced by O atoms), each of which is R QIt is arbitrarily substituted, and each R Q These are independently H, -Cl, -F, OH, CN, CF3, hydroxyl, optionally substituted linear or branched C1-C6 alkyl (e.g., methyl, ethyl), or optionally substituted linear or branched C1-C6 alkoxy; Y L1 These are, independently, combined, and NR. YL1 , O, CR YL1 R YL2 , C=O, optionally substituted linear or branched C1-C6 alkyl (any one or more C atoms are O or NR) YL1 (It is replaced by C) 1-6 Alkenes (where one or more C atoms are replaced by O), C 1-6 Alkynes (where one or more C atoms are optionally replaced by O), or optionally substituted linear or branched C1-C6 alkoxys; Q L These are 3-6 member heterocyclic rings, heterobicyclic rings, or heteroaryl rings (0-6 R). Q (which is arbitrarily substituted by) and each R Q These are independently H, or optionally substituted linear or branched C 1-6 Alkyl (for example, one or more halos, C 1-6 (Optionally substituted with alkoxyl); R YL1 , R YL2 These are H and optionally substituted linear or branched C, respectively, independently. 1-6 Alkyl (for example, one or more halos, C 1-6 It is a methyl or ethyl molecule optionally substituted with an alkoxyl; n is between 0 and 10; [ka] (This indicates a binding site to the PTM or ULM site.) Includes structures selected from.

[0142] Exemplary PTM In one aspect of this disclosure, the PTM group (also referred to as the KTM group) is a target protein, KRas or a variant thereof, for example, KRas G12C Combine.

[0143] The compositions described below include KRas binding sites (e.g., KRas) that can be used in accordance with the present invention. G12C The members of the binding site are shown as examples. These binding sites are located near ubiquitin ligases for ubiquitination and subsequent degradation of KRas proteins, for example, KRas G12C To present this feature, it is preferably linked to the ubiquitin ligase binding site (VLM) via a chemical linking group.

[0144] In a given context, the term “target protein” is used to refer to a KRas protein, a member of the RAS / MAPK pathway, which is a target protein to be ubiquitinated and degraded. In other contexts, the term “target protein” is used to refer to a KRas protein having a variant form, such as a gain-of-function KRas mutant protein or a KRas protein having one or more mutations selected from the group consisting of codon 12 missense mutations, exon 2 mutations, G12V, G12C, G12D, G12A, G13D, exon 3 mutations, codon 61 missense mutations, exon 4 mutations, G12R, Q61H, G12S, A146T, G13C, Q61R, Q61L, A146V, codon 117 missense mutations, K117N, Q61K, G12F, codon 59 missense mutations, A59T, or combinations thereof.

[0145] In any of the embodiments or examples described herein, the PTM is a KRas protein having at least one mutation that is a G12C mutation compared to the PTM that binds to wild-type KRas (e.g., KRas G12C PTM is a small molecule that selectively or preferentially binds to the KRas protein (e.g., KRas) having at least one mutation which is a G12C mutation. G12CA small molecule capable of selectively binding to a KRas protein having at least one mutation that is a G12C mutation (e.g., KRas), and its selectivity for KRas proteins having at least one mutation that is a G12C mutation is at least 1 to 60 times (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 times) compared to wild-type KRas. In any aspect or embodiment described herein, the PTM is a KRas protein having at least one mutation that is a G12C mutation (e.g., KRas G12C It is a small molecule that binds to ), and its selectivity for KRas proteins having at least one mutation that is a G12C mutation is at least 1 to 1000 times (e.g., 50, 100, 200, 300, 400, 500, 600, 700, 800, 900 times) compared to wild-type KRas.

[0146] The term "protein target site" or PTM refers to KRas or its variant forms, for example, KRas G12C This is used to describe small molecules that can bind to and be used to target proteins for ubiquitination and degradation.

[0147] The compositions described herein exemplify some of the uses of these PTMs.

[0148] In any aspect or embodiment described herein, PTM has a chemical structure: [ka] (In the formula, PTM [ka] This is the binding site for L that binds VLM or VLM to PTM; [ka] These are 6-membered aryl, 6-membered heteroaryl, or 6-membered heterocycloalkyl groups, each optionally substituted with one or two halogens (e.g., Cl, F, or Br); R PTM2 These include methyl, halogens (e.g., Cl, F, Br), amines (e.g., -NH2, -NHCH3, or -N(CH3)2), or 3- to 6-membered heterocycloalkyls (e.g., 6-membered heterocycloalkyls, heterocycloalkyls having heteroatoms selected from O and N, or [ka] It is a -C(=O)C2-C4 alkenyl arbitrarily substituted by; R PTM3A is H, phenyl, pyridinyl, isoquinoline, or naphthalene (for example, [ka] ) are, each being an OH, halogen (e.g., F, Cl, Br), amine (e.g., -NH2, -NHCH3, or -N(CH3)2), linear or branched C1-C3 haloalkyl (e.g., linear or branched C1-C3 fluoroalkyl or CF3), -R PTM3C , or optionally substituted with 1, 2, or 3 groups independently selected from linear or branched C1-C3 alkyl groups (e.g., methyl or ethyl), R PTM3C This refers to indazoles (e.g.,) optionally substituted with one, two, or three groups independently selected from OH, halogens (e.g., F, Cl, Br), or linear or branched C1-C3 alkyl groups (e.g., methyl or ethyl). [ka] (In the formula, each R PTM4D (i) is independently selected from hydrogen, C1-C3 alkyl (e.g., methyl), or halogen (e.g., F, Cl, Br); R PTM3Bis H, halogen (e.g., Cl, F, Br), or -OR PTM3C And R PTM3C It is optionally substituted with one, two, or three groups independently selected from OH, halogens (e.g., F, Cl, Br), or linear or branched C1-C3 alkyl groups (e.g., methyl or ethyl) (e.g., [ka] (In the formula, each R PTM4D (This is independently selected from hydrogen, C1-C3 alkyl (e.g., methyl), or halogen (e.g., F, Cl, Br)). PTM-I and PTM-III R PTM4A is either non-existent (or H) or one or two independently selected halogens (e.g., Cl, F, Br); R of PTM-II and PTM-IV PTM4A is either non-existent (or H) or a halogen (e.g., Cl, F, Br); R PTM4B is (1) H or absent, (2) -CH2-CH2-CN or -CH2-CN, or (3) one or two independently selected C1-C3 alkyl groups (e.g., methyl or ethyl); each X PTM (Each is either CH or N) It is represented by [this].

[0149] In any aspect or embodiment described herein, PTM has a chemical structure: [ka] [ka] (In the formula, PTM [ka] This is the binding site for L that binds VLM or VLM to PTM; RPTM3A This is an indazole optionally substituted with one or two groups independently selected from OH, methyl, and halogens (e.g., F, Cl, Br); R PTM4B is (1) absent (or H), (2) -CH2-CH2-CN or -CH2-CN, or (3) one or two independently selected C1-C3 alkyl groups (e.g., methyl or ethyl); R PTM4D is hydrogen, a C1-C3 alkyl group (e.g., methyl), or a halogen (e.g., F, Cl, Br); R PTM4C is H or a halogen (e.g., Cl, F, Br); R PTM4E is H, OH, or an amine (e.g., -NH2 or -NHCH3); R PTM4F These are hydrogen, C1-C3 alkyl (e.g., methyl), C1-C3 haloalkyl (e.g., C1-C3 fluoroalkyl or CF3), or halogen (e.g., F, Cl, Br); R PTM3B (This is an -O-indazole optionally substituted with one or two groups independently selected from OH, methyl, and halogens (e.g., F, Cl, Br)). It is represented by [this].

[0150] In any aspect or embodiment described herein, PTM has a chemical structure: [ka] (In the formula, PTM [ka] This is the binding site for L that binds VLM or VLM to PTM; R PTM3A teeth, [ka] and; R PTM4C (is H or F) It is represented by [this].

[0151] In any aspect or embodiment described herein, PTM has a chemical structure: [ka] [ka] (In the formula, R PTM4C , R PTM4D , and R PTM4E Each of these is defined independently as in any other embodiment or example described herein; PTM [ka] (This is the binding site for L that binds VLM or VLM to PTM.) It is represented by [this].

[0152] In any aspect or embodiment described herein, PTM has a chemical structure: [ka] [ka] (In the formula, R PTM4C , R PTM4D , and R PTM4E Each of these is defined independently as in any other embodiment or example described herein; PTM [ka] (This is the binding site for L that binds VLM or VLM to PTM.) It is represented by [this].

[0153] In any aspect or embodiment described herein, PTM is [ka] [ka] [ka] [ka] (In the formula, * indicates an atom that is a bonding site with a chemical linkage site or an atom shared with a chemical linkage site, PTM) [ka] (This is the binding site for L that binds VLM or VLM to PTM.) It is selected from the group consisting of the following.

[0154] In any aspect or embodiment described herein, PTM is [ka] (In the formula, [ka] is aryl, heteroaryl, cycloalkyl, or heterocycloalkyl; X PTM is C or N; W PTM This is selected from the group consisting of optionally substituted C3-C6 cycloalkyls, optionally substituted C3-C6 heteroalkyls, optionally substituted C3-C6 heterocycloalkyls, optionally substituted aryls (e.g., optionally substituted C5-C7 aryls), and optionally substituted heteroaryls (e.g., optionally substituted C5-C7 heteroaryls); R PTM1A , 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, optionally substituted -OC 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 OC 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 [ka] (For example, optionally substituted with at least one alkyl group, for example, *carbon may be optionally substituted with alkyl), optionally substituted [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] (For example, optionally substituted with at least one alkyl group, for example, *carbon may be optionally substituted with alkyl), optionally substituted [ka] (For example, optionally substituted with at least one alkyl group, for example, *carbon may be optionally substituted with alkyl), optionally substituted [ka] (For example, optionally substituted with at least one alkyl group, for example, *carbon may be optionally substituted with alkyl), optionally substituted [ka] (For example, optionally substituted with at least one alkyl group, for example, *carbon may be optionally substituted with alkyl), optionally substituted [ka] (For example, optionally substituted with at least one alkyl group, for example, *carbon may be optionally substituted with alkyl), optionally substituted [ka] (For example, optionally substituted with at least one alkyl, for example, *carbon may optionally be substituted with alkyl), where N* is the N atom of the linker (L) heterocycloalkyl (e.g., C4-C8 heterocycloalkyl); R PTM1B , 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 -OC 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, arbitrarily substituted aryl (e.g., arbitrarily substituted C5-C7 aryl), arbitrarily substituted O-aryl (e.g., arbitrarily substituted O-(C5-C7 aryl)), arbitrarily substituted heteroaryl (e.g., arbitrarily substituted C5-C7 heteroaryl), arbitrarily substituted O-heteroaryl (e.g., arbitrarily substituted O(C5-C7 heteroaryl)), arbitrarily substituted [ka] (For example, optionally substituted with at least one alkyl group, for example, *carbon may be optionally substituted with alkyl), optionally substituted [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] , arbitrarily replaced [ka] and; R PTM9 and R PTM10 These are, independently, H, an optionally substituted C1-C6 alkyl, an optionally substituted aliphatic amine, and an optionally substituted aliphatic amide; R PTM2 H, (C=O)R PTM2 , optionally substituted linear or branched alkyl; R PTM2’ This includes optionally substituted linear or branched alkyl groups, optionally substituted alkenes, and -N(R PTM8 )2, or -C(OH)2; R PTM3 These are alkyl, alkoxy, phenyl, or naphthalene compounds, each independently substituted with OH, H, or halogen; R PTM4A These are OH, H, halogens, and optionally substituted linear or branched C1-C6 alkyl groups; R PTM4BThese are OH, H, halogens, and optionally substituted linear or branched C1-C6 alkyl groups; R PTM5 This is selected from the group consisting of optionally substituted aryls, optionally substituted biaryls, optionally substituted heteroaryls, optionally substituted biheteroaryls, optionally substituted C3-C6 cycloalkyls, optionally substituted C3-C6 cycloheteroalkyls, halogens, H, optionally substituted linear or branched alkyls (e.g., optionally substituted linear or branched C1-C6 alkyls), OH, and alkoxys; R PTM8 is H or alkyl (e.g., C1 alkyl, C2 alkyl, C3 alkyl, or C4 alkyl); t is 0, 1, 2, 3, 4, 5, 6 (0, 1, 2, 3, etc.); [ka] (This indicates at least one linkage site from among linker, ULM, ULM', VLM, VLM', or a combination thereof.) It has a chemical structure represented by [the given formula / method].

[0155] In any aspect or embodiment described herein, the hetero-difunctional compound has the following chemical structure: [ka] [ka] [ka] [ka] [ka] (In the formula, [ka] These are 6-membered aryl, 6-membered heteroaryl, or 6-membered heterocycloalkyl groups, each optionally substituted with one or two halogens (e.g., Cl, F, or Br); each X PTM These are, individually, CH or N; R PTM2 These include methyl, halogens (e.g., Cl, F, Br), amines (e.g., -NH2, -NHCH3, or -N(CH3)2), or 3- to 6-membered heterocycloalkyls (e.g., 6-membered heterocycloalkyls, heterocycloalkyls having heteroatoms selected from O and N, or [ka] It is a -C(=O)C2-C4 alkenyl arbitrarily substituted by; R PTM3A is H, phenyl, pyridinyl, isoquinoline, or naphthalene (for example, [ka] ) are, each being an OH, halogen (e.g., F, Cl, Br), amine (e.g., -NH2, -NHCH3, or -N(CH3)2), linear or branched C1-C3 haloalkyl (e.g., linear or branched C1-C3 fluoroalkyl or CF3), -R PTM3C , or optionally substituted with 1, 2, or 3 groups independently selected from linear or branched C1-C3 alkyl groups (e.g., methyl or ethyl), R PTM3C This refers to indazoles (e.g.,) optionally substituted with one, two, or three groups independently selected from OH, halogens (e.g., F, Cl, Br), or linear or branched C1-C3 alkyl groups (e.g., methyl or ethyl). [ka] (In the formula, each R PTM4D (i) is independently selected from hydrogen, C1-C3 alkyl (e.g., methyl), or halogen (e.g., F, Cl, Br); RPTM3B is H, halogen (e.g., Cl, F, Br), or -OR PTM3C And R PTM3B It is optionally substituted with one, two, or three groups independently selected from OH, halogens (e.g., F, Cl, Br), or linear or branched C1-C3 alkyl groups (e.g., methyl or ethyl) (e.g., [ka] (In the formula, each R PTM4D (This is independently selected from hydrogen, C1-C3 alkyl (e.g., methyl), or halogen (e.g., F, Cl, Br)). R PTM4A is one or two independently selected halogens (e.g., Cl, F, Br); R PTM4B (1) -CH2-CH2-CN or -CH2-CN, or (2) one or two independently selected C1-C3 alkyl groups (e.g., methyl or ethyl); R 14 In any embodiment or configuration described herein, R 14 , R 14a , or R 14b As defined in; R 15 This is defined as in any embodiment or example described herein; R 16 This is defined as in any embodiment or example described herein; (o is defined as in any aspect or embodiment described herein.) It is represented by [this].

[0156] therapeutic composition The present invention further provides pharmaceutical compositions comprising a therapeutically effective amount of at least one of the difunctional compounds described herein, in combination with a pharmaceutically acceptable carrier, additive, or excipient.

[0157] In additional embodiments, this specification provides therapeutic compositions comprising an effective amount of the compounds described herein or a salt form thereof, and pharmaceutically acceptable carriers, additives or excipients, and optionally additional bioactive agents. The therapeutic compositions may be used to treat or improve disease conditions or pathologies in patients or subjects, e.g., animals such as humans, that result in targeted proteolysis and are regulated by the degradation of target proteins. In certain embodiments, the therapeutic compositions described herein may be used to achieve proteolysis for the treatment or improvement of KRas-related diseases or disorders, e.g., accumulation or hyperactivity of KRas proteins, or mutated or gain-of-function KRas proteins, misfolded KRas proteins, 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.

[0158] In alternative embodiments, this disclosure may include KRas proteins (e.g., wild-type KRas protein or KRas mutant proteins (e.g., gain-of-function KRas mutant proteins or codon 12 missense mutations, codon 12 missense mutations, exon 2 mutations, G12V, G12C, G12D, G12A, G13D, exon 3 mutations, codon 61 missense mutations, exon 4 mutations, G12R, Q61H, G12S, A146T, G13C, Q61R, Q61L, A146V, codon A method for treating a disease condition in a subject requiring it or for improving one or more symptoms of a disease or pathological condition by degrading a KRas protein having one or more mutations selected from the 117 missense mutation, K117N, Q61K, G12F, codon 59 missense mutation, A59T, or a combination thereof, comprising optionally a pharmaceutically acceptable carrier, excipient, etc. The composition provides a method which includes administering to the patient or subject in combination with an excipient, and optionally co-administering with an additional bioactive agent, and the composition is effective in treating or improving a disease or disorder or one or more symptoms in the subject. The method according to the Disclosure may be used to treat a given disease condition, pathology or symptom, including inflammatory diseases, autoimmune diseases, or cancer, by administering an effective amount of at least one compound described herein. For example, the method according to the Disclosure may be used to treat one or more of the following: accumulation or hyperactivity of KRas protein, mutated or gain-of-function KRas protein, misfolded KRas protein, 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. In any embodiment or example described herein, the method may involve administering a mutant KRas protein (e.g., KRas) to the subject before administering the composition or compound of the Disclosure to the subject. G12C This further includes identifying the patient as having ).

[0159] This disclosure further includes pharmaceutical compositions comprising pharmaceutically acceptable salts, in particular acid or base addition salts, of the compounds described herein. Acids used to prepare pharmaceutically acceptable acid addition salts of the aforementioned compounds useful in this embodiment include, among many, those that form non-toxic acid addition salts, i.e., salts containing pharmaceutically acceptable anions, such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acidic phosphate, acetate, lactate, citrate, acidic citrate, tartrate, bisulfate, succinate, maleate, fumarate, gluconate, saccharate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoic acid [i.e., 1,1'-methylene-bis-(2-hydroxy-3 naphthoate)] salts.

[0160] Pharmaceutically acceptable base addition salts may also be used to produce pharmaceutically acceptable salt forms of the compounds according to this disclosure. Chemical bases that can be used as reagents to prepare pharmaceutically acceptable base salts of the compounds are those that form non-toxic base salts with such compounds. Such non-toxic base salts include, but are not limited to, those derived from such pharmaceutically acceptable cations, e.g., alkali metal cations (e.g., potassium and sodium) and alkaline earth metal cations (e.g., calcium, zinc and magnesium), ammonium or water-soluble amine addition salts, e.g., N-methylglucamine-(meglumine), and lower alkanolammonium, as well as other base salts of pharmaceutically acceptable organic amines.

[0161] The compounds described herein may be administered orally, parenterally, or topically in single or divided doses in accordance with this disclosure. Administration of the active compound may range from continuous (intravenous) to several oral doses per day (e.g., QID), and among other routes of administration, may include oral, topical, parenteral, intramuscular, intravenous, subcutaneous, transdermal (may include osmotic enhancers), oral, sublingual, intranasal, intraocular, intrathecal, vaginal, and suppository administration. The term “parenteral,” as used herein, includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intrafocal, and intracranial injection or infusion techniques. Enteric-coated oral tablets may also be used to improve the bioavailability of the compound from the oral route of administration. The most effective form of administration depends on the pharmacokinetics of the specific drug selected, as well as the type, location, and severity of the disease, the pathology or symptoms, and the patient’s health status. The compounds according to this disclosure may also be administered as sprays, mists, or aerosols for intranasal, intratracheal, or pulmonary administration. Therefore, this disclosure also covers pharmaceutical compositions containing an effective amount of the compounds described herein, optionally combined with pharmaceutically acceptable carriers, additives, or excipients. The compounds according to this disclosure may be administered in immediate-release, intermediate-release, or sustained-release or controlled-release forms. Sustained-release or controlled-release forms are preferably administered orally, but may also be administered in suppositories and transdermal or other topical forms. Intramuscular injection in liposomal or depot formulations may also be used to control or sustain the release of the compounds at the injection site.

[0162] The compositions described herein may be formulated by conventional methods using one or more pharmaceutically acceptable carriers and may 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, e.g., human serum albumin, buffers, e.g., phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, e.g., prolamin sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and lanolin, and combinations thereof.

[0163] The sterile injectable forms of the compositions described herein may be aqueous or oily suspensions. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting and suspending agents. Sterile injectable preparations may also be sterile injectable solutions or suspensions in non-toxic, parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable vehicles and solvents that may be used include water, Ringer's solution, and isotonic sodium chloride solutions. Sterile fixative oils have also been conventionally used as solvents or suspension media. For this purpose, any sterile fixative oil, including synthetic mono or diglycerides, may be used. Fatty acids, such as oleic acid and its glyceride derivatives, as well as naturally pharmaceutically acceptable oils such as olive oil or castor oil, particularly their polyoxyethylated versions, are useful in the preparation of injectable preparations. These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants, such as Ph. Helv or similar alcohols.

[0164] The pharmaceutical compositions described herein may be administered orally in any orally acceptable dosage form, including but not limited to capsules, tablets, aqueous suspensions, or solutions. For tablets for oral use, commonly used carriers include lactose and corn starch, among those known in the art. For oral administration in capsule form, useful diluents include lactose and corn starch. Where an aqueous suspension is required for oral use, the active ingredient may be combined with emulsifiers and suspending agents. If desired, a specified sweetener, flavoring agent, or coloring agent may also be added. Lubricants, such as magnesium stearate, are also typically added.

[0165] Alternatively, the pharmaceutical compositions described herein may be administered in the form of suppositories for rectal administration. These may be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature, thereby dissolving in the rectum and releasing the drug. Such substances include cocoa butter, beeswax, and polyethylene glycol.

[0166] The pharmaceutical compositions described herein may also be administered topically. For topical use, the pharmaceutical compositions may be formulated as transdermal patches, which may be either reservoir patches or matrix patches, comprising an active compound combined with one or more carriers, buffers, and absorption enhancers, and providing continuous administration for 1 to 2 weeks.

[0167] Alternatively, the pharmaceutical compositions of this disclosure may be formulated as suitable ointments containing the active ingredient suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of this disclosure include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying waxes, and water.

[0168] Alternatively, the pharmaceutical compositions of this disclosure may be formulated as a suitable lotion or cream containing the active ingredient 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.

[0169] Alternatively, the pharmaceutical compositions of this disclosure may be formulated for ophthalmic use. For example, the pharmaceutical compositions may be formulated as an atomized suspension in isotonic pH-adjusted sterile saline, or preferably as a solution in isotonic pH-adjusted sterile saline with or without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic use, the pharmaceutical compositions may be formulated as an ointment such as petrolatum.

[0170] The pharmaceutical compositions described herein may also be administered by nasal aerosol or inhalation. Such compositions may be prepared according to techniques well known in the art of pharmaceutical formulations and may be prepared as solutions in physiological saline using benzyl alcohol or other suitable preservatives, absorption enhancers to increase bioavailability, carbon fluoride, and / or other conventional solubilizers or dispersants.

[0171] The amount of active pharmaceutical ingredient in the pharmaceutical compositions described herein, which can be combined with a carrier substance to produce a single dosage form, varies depending on the subject and disease state, the state or symptom being treated, the specific mode of administration, and the state of the subject. Preferably, the composition should be formulated to contain about 0.05 milligrams to about 750 milligrams or more, more preferably about 1 milligram to about 600 milligrams, and even more preferably about 10 milligrams to about 500 milligrams of the active ingredient, either alone or in combination with another compound according to this disclosure.

[0172] It should be understood that specific dosages and treatment regimens for any particular patient depend on a variety of factors, including the activity and bioavailability of the specific compounds used, age, weight, overall health status, sex, diet, timing of administration, rate of excretion, drug combinations, and the judgment of the treating physician and the severity of the specific disease or condition being treated.

[0173] Patients or subjects requiring therapy using compounds according to the methods described herein may be treated by administering an effective amount of the compound according to the present disclosure to the patient (subject), either alone or in combination with another known therapeutic agent, in optionally a pharmaceutically acceptable carrier or diluent, depending on a pharmaceutically acceptable salt or solvate thereof. In any embodiment or example described herein, the method involves administering a mutant KRas protein (e.g., KRas) to the subject before administering the composition or compound according to the present disclosure to the subject. G12C This further includes identifying the patient as having ).

[0174] In certain embodiments, the active compound may be combined with a pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver a therapeutically effective dose for the desired indication to the patient without causing excessive and serious toxic effects in the patient being treated. Preferred doses of the active compound for all the conditions listed herein range from about 10 nanograms per kilogram (ng / kg) to 300 milligrams per kilogram (mg / kg), preferably 0.1 to 100 mg / kg per day, more generally 0.5 to about 25 mg per kilogram of recipient / patient body weight per day. Typical topical doses range from 0.01 to 5% wt / wt in a suitable carrier.

[0175] In a given embodiment, the compound is conveniently administered in any suitable unit dosage form, including but not limited to those containing less than 1 milligram (mg), 1 mg to 3000 mg, or 5 mg to 500 mg of the active ingredient per unit dosage form. An oral dosage of approximately 25 mg to 250 mg is often convenient.

[0176] In a given embodiment, the active ingredient is administered to achieve a peak plasma concentration of the active compound, preferably about 0.00001 to 30 mmol (mM), more preferably about 0.1 to 30 micromol (μM). This can be achieved, for example, by intravenous injection of a solution or formulation of the active ingredient in saline or an aqueous medium, or by administration as a bolus of the active ingredient. Oral administration may also be appropriate to produce an effective plasma concentration of the activator.

[0177] The concentration of the active compound in a drug composition depends on the drug's absorption, distribution, inactivation, and excretion rate, as well as other factors known to those skilled in the art. It should be noted that the dosage values ​​will vary with the severity of the condition to be alleviated. It should be further understood that for any particular subject, a specific drug regimen should be adjusted over time according to individual needs and the professional judgment of the person administering or managing the composition, and that the concentration ranges shown herein are illustrative only and are not intended to limit the scope or implementation of the claimed composition. The active ingredient may be administered at once or divided into a number of smaller doses administered at different time intervals.

[0178] Oral compositions typically contain an inert diluent or food carrier. They may be encapsulated in gelatin capsules or compressed into tablets. For oral therapeutic administration, the active compound or its prodrug derivative may be incorporated with excipients and used in the form of tablets, lozenges, or capsules. Pharmaceutically compatible binders and / or adjuvants may be included as part of the composition.

[0179] Tablets, pills, capsules, lozenges, etc., may contain any of the following ingredients or compounds of similar properties: binders, e.g., microcrystalline cellulose, tragacanth gum, or gelatin; excipients, e.g., starch, or lactose; dispersants, e.g., alginic acid, Primogel, or corn starch; lubricants, e.g., magnesium stearate, or sterote; flow enhancers, e.g., colloidal silicon dioxide; sweeteners, e.g., sucrose, or saccharin; or flavoring agents, e.g., peppermint, methyl salicylate, or orange flavoring. If the dosage unit form is a capsule, it may contain a liquid carrier, e.g., fatty oil, in addition to the above types of substances. The dosage unit form may also contain various other substances that modify the physical form of the dosage unit, e.g., sugar coatings, shellac, or enteric coatings.

[0180] The active compound or a pharmaceutically acceptable salt thereof may be administered as an ingredient in elixirs, suspensions, syrups, wafers, chewing gums, etc. In addition to the active compound, the syrup may contain sucrose as a sweetener, as well as specified preservatives, pigments and colorants, and flavorings.

[0181] The active compound or a pharmaceutically acceptable salt thereof may also be mixed with other active substances that do not impair the desired effect, or substances that complement the desired effect, such as, in particular, the anticancer agents described herein. In certain preferred embodiments of this disclosure, one or more compounds according to this disclosure are co-administered with another bioactive agent, such as an anticancer agent or wound healing agent, including an antibiotic, as described elsewhere herein.

[0182] Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical applications may contain the following components: sterile diluents, e.g., water for injection, saline solution, fixative oil, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; antimicrobial agents, e.g., benzyl alcohol or methylparaben; antioxidants, e.g., ascorbic acid or sodium bisulfite; chelating agents, e.g., ethylenediaminetetraacetic acid; buffers, e.g., acetates, citrates, or phosphates; and agents for adjusting tension, e.g., sodium chloride or dextrose. Parenteral preparations may be sealed in ampoules, disposable syringes, or multi-dose vials made from glass or plastic.

[0183] When administered intravenously, the preferred carrier is physiological saline or phosphate-buffered saline (PBS).

[0184] In any embodiment or configuration, the active compound is prepared using a carrier that protects the compound from rapid excretion from the body, such as a controlled-release formulation comprising an implant and a microencapsulation delivery system. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoester, and polylactic acid may be used. Methods for preparing such formulations will become apparent to those skilled in the art.

[0185] Liposome suspensions can also be pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Patent No. 4,522,811 (which is incorporated herein by reference in its entirety). For example, a liposome formulation can be prepared by dissolving a suitable lipid(s) (e.g., stearoylphosphatidylethanolamine, stearoylphosphatidylcholine, aracadoylphosphatidylcholine, and cholesterol) in an inorganic solvent that will later be evaporated, leaving a thin film of dry lipid on the surface of a container. An aqueous solution of the active compound is then introduced into the container. The container is then rotated by hand to release the lipid material from the sides of the container, dispersing lipid aggregates and thereby forming a liposome suspension.

[0186] Treatment method

[0187] In additional embodiments, this Specified provides a therapeutic method comprising the administration of an effective amount of a compound described herein or a salt form thereof, and a pharmaceutically acceptable carrier. The therapeutic method is useful for inducing proteolysis in patients or subjects, such as animals, including humans, in order to treat or improve disease conditions, pathologies, or associated symptoms that can be treated via targeted proteolysis.

[0188] The terms “to treat,” “to treat,” and “treatment,” as used herein, refer to any action that provides a benefit to a patient to whom the Compound may be administered, including treatment of any disease condition, pathology, or symptom related to any protein to which the Compound binds. Disease conditions or pathologies, including cancer, that may be treated using the Compounds of this Disclosure are shown above.

[0189] This specification provides therapeutic methods for achieving the degradation of target proteins for the treatment or improvement of diseases, such as 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. Thus, in another embodiment, this specification provides methods for ubiquitinating / degrading target proteins in cells. In a given embodiment, the method comprises administering a bifunctional compound of the present invention. The control or reduction of specific protein levels in target cells provided by this disclosure provides treatment for a disease state, pathology, or symptom. In any embodiment or model, the method comprises administering an effective amount of the compound described herein, optionally comprising pharmaceutically acceptable excipients, carriers, adjuvants, other bioactive agents, or combinations thereof.

[0190] In additional embodiments, the Specified provides a method for treating or improving a disease, disorder or its symptoms in a subject or patient, such as an animal such as a human, comprising administering to a subject in need a composition comprising an effective amount, for example, a therapeutically effective amount, of a compound or salt form thereof described herein, and a pharmaceutically acceptable excipient, carrier, adjuvant, another bioactive agent or combination thereof, wherein the composition is effective in treating or improving the disease or disorder or its symptoms in the subject.

[0191] In any embodiment or example described herein, the method involves administering a mutant KRas protein (e.g., KRas) to a target subject before administering the composition or compound of the Disclosure. G12C This further includes identifying the patient as having ).

[0192] In another aspect, this specification provides a method for identifying the effect of compounds according to this disclosure on the degradation of a target protein in a biological system.

[0193] In another aspect, the Specified provides a process for producing a molecule that can cause degradation of KRas in cells (e.g., in vivo or in vitro), comprising: (i) providing a small molecule that binds to KRas or a variant thereof; (ii) providing an E3 ubiquitin ligase binding site (ULM), preferably a VLM as described herein; and (iii) covalently binding the small molecule from step (i) to the ULM from step (ii) via a chemical linking group (L) to form a compound that binds to both the VHL E3 ubiquitin ligase and the KRas protein and / or variant in cells, such that the VHL E3 ubiquitin ligase is in close proximity to the KRas protein bound thereto, ubiquitizes it, and then the ubiquitinated KRas is degraded.

[0194] In another aspect, the Specified herein relates to a method for detecting whether a molecule can cause degradation of a KRas protein in a cell (e.g., in vivo or in vitro), the method comprising the step of (i) providing a molecule whose ability to cause degradation of a KRas protein in a cell is detected, wherein the molecule has the structure: VLM-L-PTM (wherein VLM is a VHL E3 ubiquitin ligase binding site capable of binding to VHL E3 ubiquitin ligase in a cell, and VLM is a derivative of trans-3-hydroxyproline, as described herein, wherein both the nitrogen and carboxylic acid in trans-3-hydroxyproline are functionalized as amides; PTM is a protein targeting site which is a small molecule that binds to KRas and / or a variant KRas form thereof, wherein KRas is VHL E3 ubiquitin ligase bound to the VLM of the molecule. The present invention provides a method comprising the steps of: (ii) providing a molecule having at least one lysine residue available for ubiquitination by an E3 ubiquitin ligase (where L is a chemical linking group that covalently bonds VLM to PTM to form a molecule); (ii) incubating cells expressing the KRas protein in the presence of the molecule of step (i); and (iii) detecting whether the KRas protein has been degraded in the cells.

[0195] In any of the embodiments or models described herein, the small molecule capable of binding to KRas is a small molecule that binds to KRas. In a given embodiment, the small molecule that binds to KRas is as described herein.

[0196] In another aspect of the treatment described above, the Disclosure provides a method for treating a human patient requiring such treatment for a disease state, condition, or symptom causally related to KRas and / or KRas variant form, expression, overexpression, mutation, aggregation, accumulation, misfolding, or abnormal control, wherein the degradation of KRas produces a therapeutic effect in the patient, the method comprising administering to the patient an effective amount of the compound according to the Disclosure, optionally in combination with another bioactive agent.

[0197] A disease state, condition, or symptom may be caused by microbial substances or other exogenous substances, such as viruses, bacteria, fungi, protozoa, or other microorganisms, or may be a disease state caused by the expression, overexpression, mutation, misfolding, or abnormal regulation of proteins that lead to the disease state, condition, or symptom.

[0198] In another aspect, the Disclosure relates to a method for treating or improving at least one symptom of a disease or condition in a subject, The present invention provides a subject identified as having symptoms of a disease or condition causally related to the expression, overexpression, mutation, misfolding, or abnormal regulation of the KRas protein and / or its variant forms in the subject, wherein the symptoms of the disease or condition are treated or ameliorated by degrading the KRas protein and / or its variant forms in the cells of the subject; and a method comprising administering a compound containing a therapeutically effective amount to the subject such that the KRas protein and / or its variant forms are degraded, thereby treating or ameliorating at least one symptom of the disease or condition in the subject.

[0199] The term “disease state or pathology” is used to describe any disease state or pathology in which protein expression, overexpression, mutation, misfolding, or abnormal regulation (e.g., an increased amount of protein expressed in a patient) occurs, and the degradation of the KRas protein and / or its variant forms to reduce or stabilize the level of the KRas protein (whether mutated or not) in a patient provides a beneficial therapy or relief of symptoms for a patient in need. In the given examples, the disease state, pathology, or symptoms may be curable.

[0200] Disease conditions, pathologies, or symptoms that can be treated with the compounds of this disclosure include, for example, 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.

[0201] The term “bioactive agent” is used to describe agents other than the compounds described herein, which are used in combination with the compounds as agents having biological activity to assist in bringing about the intended therapeutic, inhibitory and / or preventive / preventive effects of the compounds. Preferred bioactive agents for use herein include those agents having pharmacological activity similar to that of the compounds used or administered, and include, for example, anticancer agents, antiviral agents (including, in particular, anti-HIV and anti-HCV agents), antibacterial agents, antifungal agents, and the like.

[0202] The term “additional anti-autoimmune disease agents” is used to describe anti-autoimmune disease therapeutic agents that may be combined with the compounds herein to treat autoimmune diseases. These agents include, for example, infliximab, tofacitinib, baricitinib, secukinumab, adalimumab etanercept, golimumab, certolizumab pepol, antiproliferative agents (e.g., mycophenolate mofetil), and corticosteroids.

[0203] The term “pharmaceutically acceptable derivative” is used throughout this specification to describe any pharmaceutically acceptable prodrug form (e.g., esters, amides, and other prodrugs) that, when administered to a patient, directly or indirectly provides the compound or its active metabolite. [Examples]

[0204] Abbreviation ACN Acetonitrile Acetic acid (ACOH) Boc tert-butoxycarbonyl DBA Dibenzylideneacetone DBU 1,8-Diazabicyclo[5.4.0]undes-7-en DCM Dichloromethane DMA (dimethylacetamide) DME (Dimethoxyethane) DMF Dimethylformamide DMSO (Dimethyl Sulfoxide) DMAC / DMA Dimethylacetamide DIEA N,N-diisopropylethylamine EDTA (Ethylenediaminetetraacetic acid) methoxy / EA ethyl acetate EtOH Ethanol FA Formic Acid HPLC (High-Pressure Liquid Chromatography) Hz (Hertz) IBX 2-Iodoxybenzoic acid LAH Lithium Aluminum Hydrogen LCMS Liquid Chromatography / Mass Spectrometry LiHMDS (Lithium Bis(Trimethylsilyl)amide) MHz (megahertz) NBS N-bromosuccinimide NCS N-chlorosuccinimide NMR nuclear magnetic resonance NMP N-methyl-2-pyrrolidone MeOH methanol MPLC (Medium Pressure Liquid Chromatography) MTBE methyl tert-butyl ether PE (Petroleum Ether) Psi = pound-force per square inch RT or rt Room temperature SFC Supercritical Fluid Chromatography TEA (Triethylamine) THF (Tetrahydrofuran) TFA (Trifluoroacetic Acid) TLC (Thin-Layer Chromatography) TMS (trimethylsilyl)

[0205] General synthetic approach

[0206] The synthesis and optimization of the bifunctional molecules described herein can be approached stepwise or modularly. For example, the identification of compounds that bind to a target protein, i.e., KRas, may involve a high- or moderate-throughput screening campaign if a suitable ligand is not readily available. The initial ligand does not typically require iterative design and optimization cycles to improve the suboptimal form identified by data from suitable in vitro and pharmacological and / or ADMET assays. Part of the optimization / SAR campaign would be to explore ligand locations that are resistant to substitution and may be suitable sites for binding to the chemical linking groups described herein. If crystallographic or NMR structural data are available, these may be used to focus on such synthetic outcomes.

[0207] Ligands for E3 ligases can be identified and optimized using a very similar method.

[0208] Using readily available PTMs and ULMs (e.g., VLMs), those skilled in the art can use known synthetic methods for their combinations with or without chemical linking groups. These chemical linking groups can be synthesized with varying compositions, lengths, and flexibility, and can be functionalized so that PTM and ULM groups can be sequentially linked to the distal ends of the linker. Thus, libraries of bifunctional molecules can be realized and profiled in in vitro and in vivo pharmacological and ADMET / PK studies. Like PTM and ULM groups, the final bifunctional molecules can be subjected to iterative design and optimization cycles to identify molecules with desired properties.

[0209] In some cases, protecting group strategies and / or functional group interconversions (FGIs) may be required to facilitate the preparation of the desired substance. Such chemical processes are well known to synthetic organic chemists, and many of them can be found in texts such as “Greene's Protective Groups in Organic Synthesis” by Peter GMWuts and Theodora W. Greene (Wiley) and “Organic Synthesis: The Disconnection Approach” by Stuart Warren and Paul Wyatt (Wiley).

[0210] Synthesis procedure

[0211] General synthesis scheme

[0212] Scheme A [ka]

[0213] Scheme B [ka]

[0214] Scheme C [ka]

[0215] Scheme 1 [ka]

[0216] Scheme 2 [ka]

[0217] Scheme 3 [ka]

[0218] Scheme 4 [ka]

[0219] Scheme 5 [ka]

[0220] Scheme 6 [ka]

[0221] Scheme 7 [ka]

[0222] Scheme 8 [ka]

[0223] Scheme 9 [ka]

[0224] Exemplary synthesis of tert-butyl(2S)-4-[6-chloro-2-(2,2-dimethoxyethoxy)-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate Step 1: Preparation of 1-benzyl 4-(tert-butyl)(R)-2-(hydroxymethyl)piperazine-1,4-dicarboxylate [ka] To a solution of tert-butyl(3R)-3-(hydroxymethyl)piperazine-1-carboxylate (10 g, 46.24 mmol, 1 e) in toluene (50 mL) and H2O (50 mL), NaHCO3 (11.66 g, 138.80 mmol, 5.40 mL, 3.00 eq) was added all at once, followed by CbzCl (11.88 g, 69.64 mmol, 9.90 mL, 1.51 eq). The resulting mixture was stirred at 0°C for 30 minutes, then at 10°C for 5 hours. The organic layer was separated and washed with water (10 mL). The aqueous phase was extracted with toluene (100 mL). The organic layers were combined, washed with water (3 x 30 mL) and brine (30 mL), dried over Na2SO4, and concentrated under reduced pressure. The resulting yellow liquid was purified by SiO2 column chromatography (25-50 SiO2 in petroleum ether) to obtain the desired product tert-butyl(2S)-4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-2-(2-oxoethoxy)quinazoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (15 g, 36.81 mmol, 80% yield, 86% purity) as a colorless liquid. LC / LC / MS(ESI) m / z: 373.1[M+Na] + . Step 2: Preparation of 1-benzyl 4-(tert-butyl)(R)-2-(((methylsulfonyl)oxy)methyl)piperazine-1,4-dicarboxylate [ka] Triethylamine (51.98 g, 513.69 mmol, 71.50 mL, 3 eq) was added all at once to a solution of tert-butyl(2S)-4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-2-(2-oxoethoxy)quinazoline-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (60 g, 171.23 mmol, 1 eq) in CH2Cl2 (500 mL). Methanesulfonyl chloride (29.42 g, 256.84 mmol, 19.88 mL, 1.5 eq) was gradually added to the solution at 0°C for 30 minutes, and the resulting mixture was stirred at 25°C for 2 hours. The reaction mixture was poured into water (500 mL), and the resulting mixture was extracted with ELISA (2 x 600 mL). The combined organic extracts were sequentially washed with saturated aqueous solution NH4Cl (500 mL), saturated aqueous solution NaHCO3 (500 mL), and brine, and then concentrated under reduced pressure to obtain the crude product 1-benzyl 4-(tert-butyl)(R)-2-(((methylsulfonyl)oxy)methyl)piperazine-1,4-dicarboxylate (70 g) as a yellow liquid. 1 H-NMR (400 MHz, CDCl3) δ 7.42 - 7.34 (m, 5H), 5.17 (s, 2H), 4.50 - 4.42 (m, 1H), 4.25 - 4.15 (m, 2H), 4.10 - 3.93 (m, 3H), 3.08 - 3.06 (m, 2H), 2.94 - 2.92 (m, 4H), 1.48(s, 9H). Step 3: Preparation of 1-benzyl 4-(tert-butyl)(S)-2-(cyanomethyl)piperazine-1,4-dicarboxylate [ka] To a solution of 1-benzyl 4-(tert-butyl)(R)-2-(((methylsulfonyl)oxy)methyl)piperazine-1,4-dicarboxylate (70 g, 163.36 mmol, 1 eq) in DMA (5 mL), KCN (16.06 g, 246.68 mmol, 10.57 mL, 1.51 eq) was added all at once, and the reaction mixture was stirred at 90°C for 8 hours. The reaction mixture was poured into ELISA (2 L), and the resulting mixture was washed with H2O (2 x 500 mL). The organic layer was washed with brine (50 mL) and then concentrated under reduced pressure. The resulting yellow liquid was purified by SiO2 column chromatography (10-20% SiO2 in petroleum ether) to obtain the desired product, 1-benzyl 4-(tert-butyl)(S)-2-(cyanomethyl)piperazine-1,4-dicarboxylate (28 g, 77.90 mmol, 47.69% yield), as a yellow liquid. LC / MS(ESI) m / z: 260.2[M-Boc+H] + . Step 4: Preparation of (S)-2-(piperazin-2-yl)acetonitrile [ka] To a solution of 1-benzyl 4-(tert-butyl)(S)-2-(cyanomethyl)piperazine-1,4-dicarboxylate (6.5 g, 25.07 mmol, 1 eq) in CH3OH (10 mL), NH4OH (4.84 g, 41.47 mmol, 5.32 mL, 30% purity, 1.65 eq) and Pd / C (1 g, 10% purity) were added in one go under N2. The suspension was degassed under vacuum, purged several times with H2, and then stirred at 10°C for 1 hour under H2 (50.53 mg, 25.07 mmol, 1 eq, 15 psi). The suspension was filtered, and the resulting clear solution was concentrated under reduced pressure. The resulting yellow liquid was reacted with HCl (4N in dioxane, 40.0 mL), and then concentrated to obtain the crude product (S)-2-(piperazin-2-yl)acetonitrile (3 g, 23.97 mmol, 95.61% yield) as a yellow liquid. Step 5: Preparation of tert-butyl(2S)-4-(7-bromo-2,6-dichloro-8-fluoroquinazolin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate [ka] To a solution of 7-bromo-2,4,6-trichloro-8-fluoroquinazoline (7.50 g, 22.69 mmol, 1 eq) and diisopropiethylamine (17.59 g, 136.13 mmol, 23.71 mL, 6 eq) in CH2Cl2 (50 mL), (S)-2-(piperazin-2-yl)acetonitrile (2.84 g, 22.69 mmol, 1 eq) in CH2Cl2 (10 mL) was added dropwise over 20 minutes, and the reaction mixture was stirred at 0°C for 30 minutes. Then, (Boc)2O (9.91 g, 45.39 mmol, 10.43 mL, 2 eq) was added little by little over 10 minutes, and the resulting mixture was stirred at 0°C for 30 minutes. The solution was concentrated under reduced pressure, and the resulting yellow solid was purified by SiO2 column chromatography (0-25% ethyl ether in petroleum ether) to obtain the desired product, tert-butyl(2S)-4-(7-bromo-2,6-dichloro-8-fluoroquinazolin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (8.1 g, 15.60 mmol, 68.75% yield), as a yellow solid. LC / MS(ESI) m / z: 520.1[M+H] + . Step 6: Preparation of tert-butyl(2S)-4-[7-bromo-6-chloro-2-(2,2-dimethoxyethoxy)-8-fluoroquinazolin-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate [ka] To a solution of tert-butyl(2S)-4-(7-bromo-2,6-dichloro-8-fluoroquinazolin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (6 g, 11.56 mmol, 1 eq) and 2,2-dimethoxyethanol (2.45 g, 23.11 mmol, 2 eq), DABCO (129.63 mg, 1.16 mmol, 127.09 uL, 0.1 eq) and Cs2CO3 (4.89 g, 15.02 mmol, 1.3 eq) was added in one step, and the reaction mixture was stirred at 50°C for 2 hours. The suspension was filtered, and the resulting clear yellow solution was concentrated under reduced pressure to obtain a yellow solid. Purification by SiO2 column chromatography (10-24% ethyl ether in petroleum ether) yielded tert-butyl(2S)-4-[7-bromo-6-chloro-2-(2,2-dimethoxyethoxy)-8-fluoroquinazolin-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (6.4 g, 10.87 mmol, 94.05% yield) as a yellow solid. Step 7: Preparation of tert-butyl(2S)-4-[6-chloro-2-(2,2-dimethoxyethoxy)-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate [ka] A mixture of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-ol (2.94 g, 10.87 mmol, 286.80 uL, 1 eq), tert-butyl(2S)-4-[7-bromo-6-chloro-2-(2,2-dimethoxyethoxy)-8-fluoroquinazolin-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (6.4 g, 10.87 mmol, 1 eq), K3PO4 (1.5 M, 21.74 mL, 3 eq), and XPhos Pd G3 (459.99 mg, 543.43 uL, 0.05 eq) in THF (15 mL) was degassed and then heated at 50 °C under an N2 atmosphere for 1 hour. The resulting suspension was filtered, and the yellow filtrate was concentrated under reduced pressure to obtain a yellow solid. Purification by prep-TLC (SiO2, 50% siRNA / petroleum ether) yielded tert-butyl(2S)-4-[6-chloro-2-(2,2-dimethoxyethoxy)-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (6.3 g, 9.66 mmol, 88.89% yield) as a yellow solid. LC / MS(ESI) m / z: 652.3[M+H] + .

[0225] Exemplary synthesis of tert-butyl(2S)-4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxo-ethoxy]quinazoline-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate Step 1: Preparation of methyl(R)-2-(benzyloxy)propanoae [ka] 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, and then thionyl chloride (39.61 g, 332.96 mmol, 24.2 mL, 3.00 eq) was added dropwise. 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) and then extracted with ethyl acetate (200 mL x 2). The combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 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 (400MHz, CDCl3) 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). Step 2: Preparation of (R)-2-(benzyloxy)propanal [ka] A solution of methyl(2R)-2-benzyloxypropanoate (20.96 g, 107.92 mmol, 1.00 eq) in dichloromethane (200 mL) was cooled to -78°C, and then diisobutylaluminum hydride (1 M, 110 mL, 1.00 eq) was added dropwise. The mixture was then stirred at -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) and then extracted with dichloromethane (100 mL x 2). The combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. Compound (2R)-2-benzyloxypropanal (15.70 g) was obtained as a colorless oil. LC / MS (ESI) m / z: 181.1 [M+17] + ; 1H-NMR (400MHz, CDCl3) 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). Step 3: Preparation of (R)-(((1,1-dimethoxypropan-2-yl)oxymethyl)benzene [ka] 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; pyridine (450 mg, 1.79 mmol, 0.02 eq) was added. The mixture was stirred at 25°C for 2 hours. The reaction mixture was diluted with water (100 mL) and then extracted with ethyl acetate (100 mL x 2). The combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the 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 (400MHz, CDCl3) 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). Step 4: Preparation of (R)-1,1-dimethoxypropan-2-ol [ka] To a solution of [(1R)-2,2-dimethoxy-1-methylethoxy]methylbenzene (9.00 g, 42.80 mmol, 1.00 eq) in methanol (80 mL), palladium-supported activated carbon (500 mg, 5% purity) and palladium hydroxide (500 mg, 5% purity) were added under nitrogen gas. The suspension was degassed under vacuum and purged several times with hydrogen gas. The mixture was stirred at 60°C for 8 hours under hydrogen gas (15 psi). The reaction mixture was filtered through Celite and concentrated under reduced pressure to obtain the residue. Compound (2R)-1,1-dimethoxypropan-2-ol (4.1 g) was obtained as a colorless oil. 1 H-NMR (400MHz, CDCl3) 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). Step 5: Preparation of tert-butyl(2S)-4-[7-bromo-6-chloro-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-8-fluoroquinazolin-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate [ka] To a solution of tert-butyl(2S)-4-(7-bromo-2,6-dichloro-8-fluoroquinazolin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (500 mg, 0.96 mmol, 1.00 eq) and (2R)-1,1-dimethoxypropan-2-ol (231 mg, 1.93 mmol, 2.00 eq) in CH3CN (5 mL), 1,4-diazabicyclo[2.2.2]octane (11 mg, 0.01 mmol, 0.10 eq) and Cs2CO3 (408 mg, 1.25 mmol, 1.30 eq) were added, and the reaction mixture was stirred at 45°C for 2 hours. The solvent was removed under reduced pressure, and the resulting residue was purified by SiO2 column chromatography (15-25% ethyl ether in petroleum ether) to obtain tert-butyl(2S)-4-[7-bromo-6-chloro-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-8-fluoroquinazoline-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (250 mg, 0.41 mmol, 43% yield) as a pale yellow solid. LC / MS(ESI) m / z: 604.2[M+H] + . Step 6: Preparation of tert-butyl(2S)-4-[6-chloro-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate [ka] A solution of tert-butyl(2S)-4-[7-bromo-6-chloro-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-8-fluoroquinazolin-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (250 mg, 0.41 mmol, 1.00 eq) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-ol (129 mg, 0.48 mmol, 1.15 eq) in THF (3.0 mL) is mixed with [2-(2-aminophenyl)phenyl]palladium (1+); dicyclohexyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphan; methanesulfonate (35 mg, 0.04 mmol, 0.10 eq) and K3PO4 (1.5 M, 0.8 mL). 3.00 eq) was added, the reaction mixture was degassed with N2 gas, and stirred at 45°C for 9 hours. Then water (30 mL) was added, and the resulting mixture was extracted with ELISA (2 x 50 mL). The combined organic extract was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated. The resulting residue was purified by prep-thin-layer chromatography (ELISA:petroleum ether = 2:3) to obtain tert-butyl(2S)-4-[6-chloro-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (180 mg, 0.27 mmol, 65% yield) as a pale red solid. LC / MS(ESI) m / z: 666.4[M+H] + . Step 7: Preparation of tert-butyl(2S)-4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxo-ethoxy]quinazoline-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate [ka] To a solution of tert-butyl(2S)-4-[6-chloro-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (130 mg, 0.20 mmol, 1.00 eq) in CH2Cl2 (0.5 mL), TFA (1.54 g, 13.51 mmol, 1.0 mL, 69.21 eq) was added, and the reaction mixture was stirred at 25 °C for 12 hours. The solvent was removed under reduced pressure, and the mixture was dried in vacuum. The resulting substance was incorporated into a mixture of THF (1.5 mL) and H2O (1.5 mL). Next, NaHCO3 (292 mg, 3.47 mmol, 20.00 eq) was added gradually, followed by di-tert-butyl dicarbonate (57 mg, 0.26 mmol, 1.50 eq), and the reaction mixture was stirred at 25°C for 12 hours. The mixture was diluted with H2O (20 mL) and extracted with siRNA (2 x 20 mL). The combined organic extract was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated. The obtained residue was purified by prep-thin-layer chromatography (CH2Cl2:CH3OH=30:1) to obtain tert-butyl(2S)-4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxo-ethoxy]quinazoline-4-yl]-2-(cyanomethyl)piperazine-1-carboxylate (30 mg, 0.05 mmol, 28% yield) as a yellow solid. LC / MS(ESI) m / z: 620.3[M+H] + and 720.3[M+Boc+1] + .

[0226] Exemplary synthesis of tert-butyl 4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-2-(((R)-1-oxopropan-2-yl)oxy)quinazoline-4-yl)piperazine-1-carboxylate 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 [ka] 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 mmol, 0.1 eq) were placed in a microwave tube in acetonitrile (10 mL). The sealed tube was heated under microwave conditions at 100°C for 2 hours. The reaction mixture was filtered, and the filtrate was concentrated under vacuum to obtain the residue. The residue was purified by silica gel column chromatography (0-15% ethyl acetate in petroleum ether) to obtain tert-butyl 4-[7-bromo-6-chloro-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-8-fluoroquinazolin-4-yl]piperazine-1-carboxylate (706 mg, 1.10 mmol, 22% yield, 87% purity) as a yellow solid. LC / MS(ESI) m / z: 565.1[M+1] + . Step 6: Preparation of tert-butyl 4-(6-chloro-2-(((R)-1,1-dimethoxypropan-2-yl)oxy)-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazoline-4-yl)piperazine-1-carboxylate [ka] 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) were added to a solution of tert-butyl 4-[7-bromo-6-chloro-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-8-fluoroquinazolin-4-yl]piperazine-1-carboxylate (608 mg, 1.08 mmol, 1 eq) and 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). The reaction mixture was degassed, charged with nitrogen three times, and then heated at 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 under vacuum to obtain the residue. The residue was purified by flash silica gel chromatography (0-60% ethyl acetate in petroleum ether) to obtain the crude product (600 mg). This crude product was purified by preparative reverse-phase HPLC. The collected fraction was concentrated under vacuum 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 x 2). The combined organic layers were dried over sodium sulfate and then concentrated under vacuum to obtain 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) as a pale yellow solid. LC / MS(ESI) m / z: 627.2[M+1] + . Step 7: Preparation of (2R)-2-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-4-(piperazine-1-yl)quinazoline-2-yl)oxy)propanal [ka] A solution of 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, 0.64 mmol, 1 eq) in dioxane (20 mL) was mixed with 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 under vacuum to obtain (2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1-yl-quinazolin-2-yl]oxypropanal (330 mg, hydrochloride) as a pale yellow, rubbery substance. LC / MS (ESI) m / z: 481.1 [M+1] + . Step 8: Preparation of tert-butyl 4-(6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-2-(((R)-1-oxopropan-2-yl)oxy)quinazoline-4-yl)piperazine-1-carboxylate [ka] A mixture of (2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1-ylquinazolin-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) was cooled to 0°C. Then saturated aqueous sodium sulfate (322 mg, 3.83 mmol, 6 mL, 6 eq) was 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 under vacuum to obtain the residue. The residue was purified by silica gel column chromatography with dichloromethane (50 mL) and then ethyl acetate (40 mL) to obtain 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 (380 mg) as a pale yellow solid. LC / MS (ESI) m / z: 581.2 [M+1] + ; 1 H-NMR (400MHz, CDCl3) 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).

[0227] Exemplary synthesis 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 Step 1: Preparation of tert-butyl(S)-(1-(4-bromophenyl)ethyl)carbamate [ka] In a 250 mL round-bottom flask, (1S)-1-(4-bromophenyl)ethane-1-amine (10.0 g, 49.98 mmol, 1.00 equivalent), triethylamine (10.0 g, 99.01 mmol, 2.00 equivalent), and di-tert-butyl dicarbonate (13.0 g, 59.63 mmol, 1.20 equivalent) were added in 100 mL of dichloromethane. The resulting solution was stirred at room temperature for 2 hours. The resulting mixture was concentrated under vacuum. The residue was applied to a silica gel column with ethyl acetate / petroleum ether (1:10). This yielded 15.0 g of tert-butyl N-[(1S)-1-(4-bromophenyl)ethyl]carbamate as a white solid. Step 2: Preparation of tert-butyl(S)-(1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)carbamate [ka] In a 250 mL round-bottom flask purged and maintained under an inert nitrogen atmosphere, a solution of tert-butyl N-[(1S)-1-(4-bromophenyl)ethyl]carbamate (15.0 g, 49.97 mmol, 1.00 equivalent), 4-methyl-1,3-thiazole (9.9 g, 99.84 mmol, 2.00 equivalent), potassium acetate (9.8 g, 99.86 mmol, 2.00 equivalent), and palladium(II) acetate (112.5 mg, 0.50 mmol, 0.01 equivalent) were added. The resulting solution was stirred at 120°C for 2 hours. 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 under vacuum. The residue was applied to a silica gel column in ethyl acetate / petroleum ether (1:5). This yielded 7.5 g (47%) of tert-butyl N-[(1S)-1-[4-(4-methyl-1,3-thiazole-5-yl)phenyl]ethyl]carbamate as a white solid. LC / MS(ESI) m / z: 319.13[M+Na] + . Step 3: Preparation of (S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethane-1-amine hydrochloride [ka] A solution of tert-butyl N-[(1S)-1-[4-(4-methyl-1,3-thiazole-5-yl)phenyl]ethyl]carbamate (7.5 g, 23.55 mmol, 1.00 equivalent) in methanol (20 mL) was placed in a 100 mL round-bottom flask, and hydrochloric acid (gas) was bubbling in at room temperature. The resulting solution was stirred at room temperature for 2 hours. The resulting mixture was concentrated under vacuum. This yielded 4.4 g (86%) of (1S)-1-[4-(4-methyl-1,3-thiazole-5-yl)phenyl]ethane-1-amine as a white solid. Step 4: Preparation of tert-butyl(2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carboxylate [ka] In a 100 mL round-bottom flask, (2S,4R)-1-[(tert-butoxy)carbonyl]-4-hydroxypyrrolidine-2-carboxylic acid (4.7 g, 20.32 mmol, 1.00 equivalent), N-ethyl-N-isopropylpropan-2-amine (7.8 g, 60.35 mmol, 3.00 equivalent), o-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.5 g, 30.26 mmol, 1.50 equivalent), and (1S)-1-[4-(4-methyl-1,3-thiazole-5-yl)phenyl]ethane-1-amine (4.4 g, 20.15 mmol, 1.00 equivalent) were added in 20 mL of N,N-dimethylformamide. The resulting solution was stirred at room temperature for 12 hours. 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, dried under reduced pressure in an oven, and concentrated under vacuum. The residue was applied to a silica gel column with ethyl acetate / petroleum ether (1:1). This yielded 5.0 g (57%) of tert-butyl(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methyl-1,3-thiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carboxylate as a yellow solid. LC / MS (ESI) m / z: 432.15 [M+1] + . Step 5: Preparation of (2S,4R)-4-hydroxy-N-((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide hydrochloride [ka] A solution of tert-butyl(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methyl-1,3-thiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carboxylate (5.0 g, 11.59 mmol, 1.00 equivalent) in methanol (200 mL) was placed in a 500 mL round-bottom flask, and hydrochloric acid (gas) was then bubbling over the reaction mixture at room temperature for 2 hours. The resulting mixture was concentrated under vacuum. This yielded 3.2 g (83%) of (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide as a red solid. Step 6: Preparation of tert-butyl((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)carbamate [ka] (2S)-2-[(tert-butoxy)carbonyl]amino-3,3-dimethylbutanoic acid (2.0 g, 8.65 mmol, 0.99 equivalents) in N,N-dimethylformamide (30 mL) was added to a 25 mL round-bottom flask. N-ethyl-N-isopropylpropan-2-amine (3.4 g, 3.00 equivalents), o-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (5.0 g, 1.50 equivalents), (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide hydrochloride (3.2 g, 8.70 mmol, 1.00 equivalent). The resulting solution was stirred at room temperature for 12 hours. The solution was extracted with ethyl acetate (60 mL x 3) and washed with water (100 mL x 2). The organic layers were combined, dried, and concentrated under vacuum. The residue was applied to a silica gel column in ethyl acetate / petroleum ether (1:3). This yielded 4.0 g (84%) of tert-butyl N-[(2S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methyl-1,3-thiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate as a yellow solid. LC / MS (ESI) m / z: 545.30[M+1] + . Step 7: Preparation 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 [ka] A solution of tert-butyl N-[(2S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methyl-1,3-thiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-yl]-3,3-dimethyl-1-oxobutan-2-yl]carbamate (4.0 g, 7.34 mmol, 1.00 equivalent) in methanol (30 mL) was placed in a 100 mL round-bottom flask, and hydrochloric acid (gas) was then bubbling over the reaction mixture at room temperature for 2 hours. The resulting mixture was concentrated under vacuum. This yielded 3.5 g of (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide hydrochloride as a yellow solid. LC / MS(ESI)m / z:445.05[M+1] + ; 1 H-NMR (400MHz, DMSO-d6) δ 8.99 (s, 1 H), 8.57-8.55 (d, J = 7.8 Hz, 1 H), 8.01 (b, 3 H), 7.46-7.43 (d, J = 8.4 Hz, 2 H), 7.39-7.37 (d, J = 8.4 Hz, 2 H), 4.98-4.90 (m, 1 H), 4.57-4.51 (m, 1 H), 4.34 (b, 1 H), 3.94-3.92 (m, 1 H), 3.69-3.66 (m, 1 H), 3.53-3.49 (m, 1 H), 2.52 (s, 3 H), 2.10-2.07 (m, 1 H), 1.83-1.81 (m, 1H), 1.40-1.30 (m, 3H), 1.03 (s, 9H).

[0228] Exemplary synthesis of (2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazole-5-yl)benzyl)-1-(3-methyl-2-(3-methylisoxazole-5-yl)butanoyl)pyrrolidine-2-carboxamide Step 1: Preparation of 2-(3-methylisoxazole-5-yl)acetic acid [ka] 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) was added dropwise under nitrogen at -78 °C. The mixture was stirred at -55 °C for 30 minutes, then carbon dioxide was bubbling into the mixture for 30 minutes, and the mixture was stirred at 25 °C for 1 hour. The mixture was quenched with saturated ammonium chloride solution (50 mL) and extracted with ethyl acetate (50 mL). The aqueous phase was adjusted to pH=2 with aqueous hydrochloric acid solution (2 M), and the mixture was extracted with ethyl acetate (50 mL, 3 times). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain 2-(3-methylisoxazole-5-yl)acetic acid (10 g, 70.86 mmol, 46% yield) as a brown solid. 1 H-NMR (400MHz, DMSO-d6) δ 12.74 (brs, 1H), 6.24 (s, 1H), 3.83 (s, 2H), 2.20 (s, 3H). Step 2: Preparation of methyl 2-(3-methylisoxazole-5-yl)acetate [ka] A solution of 2-(3-methylisoxazole-5-yl)acetic acid (10 g, 70.86 mmol, 1 eq) in methanol (100 mL) was mixed with 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 obtain the crude product. This crude product was diluted with ethyl acetate (200 mL), washed with water (200 mL), then saturated sodium bicarbonate aqueous solution (50 mL), then brine (50 mL), the organic phase was dried anhydrous, filtered, and the filtrate was condensed to obtain methyl 2-(3-methylisoxazole-5-yl)acetate (10 g, 64.45 mmol, 91% yield) as a brown oil. 1 H-NMR (400MHz, CDCl3) δ 6.11 (s, 1H), 3.80 (s, 2H), 3.76 (s, 3H), 2.30 (s, 3H). Step 3: Preparation of methyl 3-methyl-2-(3-methylisoxazole-5-yl)butanoate [ka] To a solution of methyl 2-(3-methylisoxazole-5-yl)acetate (10 g, 64.45 mmol, 1 eq) in tetrahydrofuran (100 mL), sodium hydride (3.87 g, 96.68 mmol, 60% purity, 1.5 eq) was added at 0°C, followed by the addition of 2-iodopropane (13.15 g, 77.34 mmol, 7.74 mL, 1.2 eq) at 0°C. The mixture was 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 an aqueous hydrochloric acid solution (1 M, 300 mL), extracted with ethyl acetate (200 mL, 3 times), the organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain methyl 3-methyl-2-(3-methylisoxazole-5-yl)butanoate (13 g) as a brown oil. Step 4: Preparation of 3-methyl-2-(3-methylisoxazole-5-yl)butanoic acid [ka] To a solution of methyl 3-methyl-2-(3-methylisoxazole-5-yl)butanoate (12.7 g, 64.39 mmol, 1 eq) in methanol (90 mL) and water (60 mL), sodium hydroxide (12.88 g, 321.96 mmol, 5 eq) was added, and the mixture was stirred at 25°C for 2 hours. The mixture was concentrated to remove methanol, the residue was then diluted with water (200 mL), extracted with ethyl acetate (200 mL), the aqueous phase was adjusted to pH=3 with aqueous hydrochloric acid (2 M), the mixture was then extracted with dichloromethane (200 mL, 3 times), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product as a brown oil. This crude product was purified by flash prep-HPLC to remove the acetonitrile fraction, 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 obtain the product 3-methyl-2-(3-methylisoxazole-5-yl)butanoic acid (7.5 g, 40.94 mmol, 63% yield) as a white solid. 1 H-NMR (400MHz, DMSO-d6) 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). Step 5: Preparation of 2-hydroxy-4-(4-methylthiazole-5-yl)benzonitrile [ka] 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 under nitrogen for 6 hours. The mixture was quenched with water (500 mL), and the aqueous phase was extracted with ethyl acetate (300 mL x 3). The combined organic phase was washed with brine (200 mL, twice), dried over anhydrous sodium sulfate, filtered, concentrated under vacuum, and then methyl tert-butyl ether (500 mL) was added to the mixture, and the organic phase was washed with water (100 mL) and brine (100 mL, twice). The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1 to 1 / 1). Compound 2-hydroxy-4-(4-methylthiazole-5-yl)benzonitrile (11 g, 50.87 mmol, 67% yield) was obtained as a yellow solid. Step 6: Preparation of 2-(aminomethyl)-5-(4-methylthiazole-5-yl)phenol [ka] A solution of 2-hydroxy-4-(4-methylthiazole-5-yl)benzonitrile (11 g, 50.87 mmol, 1 eq) in tetrahydrofuran (150 mL) was mixed with lithium aluminum hydride (7.72 g, 203.46 mmol, 4 eq) at 0°C, and the mixture was stirred at 50°C for 3 hours. The mixture was quenched with water (8 mL) at 0°C, then 15% aqueous sodium hydroxide solution (8 mL), then water (8 mL), and anhydrous sodium sulfate (30 g) were added. The mixture was stirred at 25°C for 30 minutes, filtered, and the solid was added to dichloromethane / methanol (4 / 1, 50 mL). The mixture was stirred at 25°C for 1 hour, filtered, and the combined filtrate was concentrated to obtain 2-(aminomethyl)-5-(4-methylthiazole-5-yl)phenol (7 g, 31.78 mmol, 62% yield) as a brown solid. 1H-NMR (400MHz, DMSO-d6) 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). Step 7: Preparation of tert-butyl(2S,4R)-4-hydroxy-2-((2-hydroxy-4-(4-methylthiazole-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate [ka] To a solution of 2-(aminomethyl)-5-(4-methylthiazole-5-yl)phenol (7 g, 31.78 mmol, 1 eq) and (2S,4R)-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), followed by HATU (13.29 g, 34.95 mmol, 1.1 eq), was added, and the mixture was stirred at 25°C for 2 hours. Additional (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) were added, and the mixture was stirred at 25°C for 5 hours. The mixture was diluted with water (300 mL), extracted with ethyl acetate (300 mL, twice), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product as a brown oil. This crude product was dissolved in tetrahydrofuran / water (1 / 2, 150 mL), 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), the pH was adjusted to 7 with aqueous hydrochloric acid (0.5 M), the mixture was extracted with ethyl acetate (300 mL, twice), the organic phase was dried over 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% metaneal in dichloromethane) to obtain tert-butyl(2S,4R)-4-hydroxy-2-[[2-hydroxy-4-(4-methylthiazole-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] + . Step 8: Preparation of (2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazole-5-yl)benzyl)pyrrolidine-2-carboxamide [ka] To a solution of tert-butyl(2S,4R)-4-hydroxy-2-[[2-hydroxy-4-(4-methylthiazole-5-yl)phenyl]methylcarbamoyl]pyrrolidine-1-carboxylate (6.9 g, 15.92 mmol, 1 eq) in methanol (30 mL), hydrogen chloride / 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 obtain the product as a yellow solid, and this crude product was triturated with ethyl acetate and petroleum ether (1:1, 20 mL). The mixture was filtered, and the solid was dried by rotary evaporator to obtain the product (2S,4R)-4-hydroxy-N-[[2-hydroxy-4-(4-methylthiazole-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (4.83 g, 13.06 mmol, 82% yield, hydrochloric acid) as a yellow solid. 1 H-NMR (400MHz, DMSO-d6) 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). Step 9: Preparation of (2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazole-5-yl)benzyl)-1-(3-methyl-2-(3-methylisoxazole-5-yl)butanoyl)pyrrolidine-2-carboxamide [ka] To a solution of (2S,4R)-4-hydroxy-N-[[2-hydroxy-4-(4-methylthiazole-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 the addition of 3-methyl-2-(3-methylisoxazole-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 for 2 hours. The mixture was diluted with water (200 mL), extracted with ethyl acetate (300 mL, twice), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by prep-HPLC to remove the acetonitrile fraction, 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 obtain the product (2S,4R)-4-hydroxy-N-[[2-hydroxy-4-(4-methylthiazole-5-yl)phenyl]methyl]1-[3-methyl-2-(3-methylisoxazole-5-yl)butanoyl]pyrrolidine-2-carboxamide (4.0 g, 8.02 mmol, 61% yield) as a white solid. 1 H-NMR (400MHz, CD3OD) δ 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).

[0229] Exemplary 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 Step 1: Preparation of benzyl(S)-2-(cyanomethyl)piperazine-1-carboxylate [ka] Trifluoroacetic acid (3.08 g, 27.00 mmol, 10.79 eq) was gradually added 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). The solution was stirred at 10°C for 2 hours. The solution was concentrated under reduced pressure to obtain benzyl(2S)-2-(cyanomethyl)piperazine-1-carboxylate (911 mg, 2.44 mmol, 97% yield, trifluoroacetate) as a yellow liquid. Step 2: 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 [ka] To a solution of benzyl(2S)-2-(cyanomethyl)piperazine-1-carboxylate (646 mg, 2.49 mmol, 1.00 eq, trifluoroacetate) and diisopropylethylamine (1.29 g, 9.96 mmol, 4.00 eq) in dimethyl sulfoxide (20 mL), tert-butyl 2,4-dichloro-5,6-dihydropyrido[3,4-d]pyrimidine-7(8H)-carboxylate (758 mg, 2.49 mmol, 1.00 eq) was added all at once. 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 and collected, washed with water (50 mL x 2) and brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a yellow liquid. The yellow liquid was purified by column chromatography (silicon dioxide, petroleum ether / ethyl acetate = 10 / 1 to 1 / 1) to obtain tert-butyl 4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-2-chloro-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-7-carboxylate (1.10 g, 2.09 mmol, 84% yield) as a yellow liquid. LC / MS(ESI) m / z: 527.1[M+1] + ; 1 H-NMR (400MHz, CDCl3) 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).

[0230] Exemplary synthesis of (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-(4-piperidyloxy)ethoxy]acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide Step 1: Preparation of tert-butyl 4-(2-(2-ethoxy-2-oxoethoxy)ethoxy)piperidine-1-carboxylate [ka] Diacetoxyrhodium (90 mg, 0.41 mmol, 0.05 eq) was added to a solution of tert-butyl 4-(2-hydroxyethoxy)piperidine-1-carboxylate (2 g, 8.15 mmol, 1 eq) in dichloromethane (20 mL). Ethyl 2-diazoacetate (2.79 g, 24.46 mmol, 3 eq) was added at 0°C, and the mixture was stirred at 25°C for 12 hours. The reaction mixture was concentrated under vacuum. The crude product was purified by flash silica gel chromatography (petroleum ether:ethyl acetate = 9:1~3:1). 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 (400MHz, CDCl3) 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). Step 2: Preparation of 2-(2-((1-(tert-butoxycarbonyl)piperidine-4-yl)oxy)ethoxy)acetic acid [ka] Lithium hydroxide monohydrate (405 mg, 9.66 mmol, 2 eq) was added 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), tetrahydrofuran (3 mL), and water (3 mL). 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 1 M hydrochloric acid, and the aqueous phase was extracted with dichloromethane and methanol (10:1, 30 mL x 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. 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. Step 3: Preparation of tert-butyl 4-(2-(2-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)ethoxy)piperidine-1-carboxylate [ka] 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-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2- in N,N-dimethylformamide (10 mL) To a solution of carboxamide (600 mg, 1.35 mmol, 1 eq), hydroxybenzotriazole (274 mg, 2.03 mmol, 1.50 eq), 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) were added. 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 over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (dichloromethane:methanol = 1:0 to 20:1). The compound tert-butyl 4-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-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, 70% yield) was obtained as a yellow oil. LC / MS (ESI) m / z: 730.4[M+1] + . Step 4: Preparation of (2S,4R)-1-((S)-3,3-dimethyl-2-(2-(2-(piperidine-4-yloxy)ethoxy)acetamide)butanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide [ka] A mixture of tert-butyl 4-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-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 removed again, and the mixture was dried under 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-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (470 mg, 705.42 ml, 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, washed with dichloromethane (15 mL), the filtrate was concentrated, and dried under vacuum. The compound (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-(4-piperidyloxy)ethoxy]acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (295 mg, 0.47 mmol, 66% yield) was obtained as a pale yellow solid.

[0231] (2S,4R)-1-((S)-2-(2-(2-((1-((R)-2-(((S)-4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazoline-2-yl)oxy)propyl)piperidine-4-yl)oxy)ethoxy)acetamide)-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide and (2 Exemplary synthesis of S,4R)-1-((S)-2-(2-(2-((1-((R)-2-(((R)-4-(4-acryloylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)quinazoline-2-yl)oxy)propyl)piperidine-4-yl)oxy)ethoxy)acetamide)-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide Step 1: 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-methylthiazole-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)ethoxy)piperidine-1-yl)propan-2-yl)oxy)-7-(3-hydroxynaphthalene-1-yl)quinazoline-4-yl)piperazine-1-carboxylate [ka] 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-piperidine To a solution of [(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (78 mg, 0.12 mmol, 1.2 eq), acetic acid (12 mg, 0.20 mmol, 2 eq) was added, followed by sodium borohydride cyanohydride (19 mg, 0.31 mmol, 3 eq) at 0°C. The mixture was stirred at 25°C for 3 hours. The mixture was concentrated under vacuum. The mixture was purified by prep-TLC (dichloromethane:methanol = 10:1) to obtain the product. The compound tert-butyl4-[6-chloro-8-fluoro-2-[(1R)-2-[4-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-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)quinazoline-4-yl]piperazine-1-carboxylate (63 mg) was obtained as a yellow solid. LC / MS (ESI) m / z: 1194.2 [M+1] + . Step 2: Preparation of (2S,4R)-1-((2S)-2-(2-(2-((1-((2R)-2-((6-chloro-8-fluoro-7-(3-hydroxynaphthalene-1-yl)-4-(piperazine-1-yl)quinazoline-2-yl)oxy)propyl)piperidine-4-yl)oxy)ethoxy)acetamide)-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide [ka] Trifluoroacetic acid (770 mg, 6.75 mmol, 0.5 mL, 132.28 eq) was added 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-methylthiazole-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)quinazoline-4-yl]piperazine-1-carboxylate (61 mg, 0.05 mmol, 1 eq) in dichloromethane (2 mL). The mixture was stirred at 25°C for 0.5 hours. The mixture was concentrated under vacuum. Compound (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1-ylquinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (61 mg, 0.05 mmol, 99% yield, trifluoroacetate) was obtained as a yellow oil. Step 3: Preparation 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)quinazoline-2-yl)oxy)propyl)piperidine-4-yl)oxy)ethoxy)acetamide)-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide [ka] (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1-ylquinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)- To a solution of 1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (61 mg, 0.05 mmol, 1 eq, trifluoroacetate), 2,6-lutidine (54 mg, 0.50 mmol, 10 eqs) was added, followed by the addition of prop-2-enoyl chloride (4 mg, 0.045 mmol, 0.9 eqs) in dichloromethane (4 mL) at -65°C. The mixture was stirred at -65°C for 10 minutes. Water (10 mL) was added. The aqueous phase was extracted with dichloromethane (15 mL x 3). The combined organic phase was concentrated under vacuum. The residue was purified by preparative 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-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (19 mg, 0.02 mmol, 31% yield, 99% purity, formate) was obtained as a white solid. LC / MS (ESI) m / z: 574.8 [M / 2+1] + ; 1H-NMR (400MHz, DMSO-d6) 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 (br d, J=8.3 Hz, 2H), 7.06 (br d, 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)。

[0232] Step 4: Separation of atropisomers 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-methylthiazole-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)quinazoline-4-yl]piperazine-1-carboxylate [ka] The atropisomers 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-methylthiazole-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)quinazoline-4-yl]piperazine-1-carboxylate (105 mg, 0.088 mmol, 1 eq) were separated by SFC (60% isopropanol in 0.1% NH4OH). Step 5: Preparation of (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1-ylquinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] 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-methylthiazole-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)quinazoline-4-yl]piperazine-1-carboxylate (56 mg, 0.047 mmol, 1 eq), trifluoroacetic acid (383 mg, 3.36 mmol, 71.72 eq) was added, and the reaction mixture was stirred at 20°C for 1 hour. The reaction mixture was concentrated under vacuum to obtain (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1-ylquinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (56 mg, 0.046 mmol, 99% yield, TFA salt) as a yellow, rubbery substance. Step 6: Preparation of (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1-ylquinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1- To a solution of [4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (56 mg, 0.046 mmol, 1 eq, TFA salt) and 2,6-lutidine (99 mg, 0.093 mmol, 20 eqs), prop-2-enoyl chloride (3.77 mg, 0.042 mmol, 0.9 eq) was added in CH2Cl2 (0.34 mL), and the reaction mixture was stirred at -70°C for 10 minutes. The reaction mixture was diluted with CH2Cl2 (30 mL) and water (20 mL). The organic layer was separated, and the aqueous layer was further extracted with CH2Cl2 (10 mL). The combined organic extract was dried over Na2SO4 and concentrated. The obtained residue was purified by preparative reverse-phase HPLC (36-56% CH3CN in water (0.1% TFA)) to obtain (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (26.5 mg, 0.021 mmol, 45% yield, 99% purity, TFA salt) as a colorless, rubbery substance. LC / MS(ESI)m / z:1148.5[M+H] + . 1H-NMR (400 MHz, DMSO-d6) δ 10.12 (s, 1H), 10.20 - 9.88 (m, 1H), 9.18 (s, 1H), 9.01 - 8.96 (m, 1H), 8.82 - 8.68 (m, 1H), 8.44 - 8.35 (m, 1H), 8.09 - 8.04 (m, 1H), 7.87 - 7.78 (m, 1H), 7.49 - 7.38 (m, 4H), 7.38 - 7.32 (m, 3H), 7.31 - 7.28 (m, 1H), 7.26 - 7.16 (m, 2H), 7.10 - 7.05 (m, 1H), 6.89 - 6.77 (m, 1H), 6.15 (s, 1H), 5.80 - 5.73 (m, 1H), 5.70 - 5.58 (m, 1H), 4.89 (br t, J = 7.0 Hz, 1H), 4.54 (dd, J = 7.0, 9.5 Hz, 1H), 4.44 - 4.39 (m, 1H), 4.32 - 4.22 (m, 2H), 3.96 (br d, J = 5.3 Hz, 3H), 3.91 (br s, 2H), 3.87 (br d, J = 1.0 Hz, 2H), 3.80 (br s, 2H), 3.66 (br s, 1H), 3.63 - 3.49 (m, 10H), 3.42 - 3.34 (m, 1H), 3.27 - 2.96 (m, 2H), 2.45 (s, 3H), 2.18 - 1.92 (m, 3H), 1.91 - 1.60 (m, 2H), 1.57 - 1.41 (m, 1H), 1.40 - 1.31 (m, 6H), 0.92 (br d, J = 4.0 Hz, 9H)。 The following opposite atropisomer was obtained by a similar method: (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] (TFA salt, colorless, rubbery substance). LC / MS (ESI) m / z: 1148.5 [M+1] + . 1H-NMR (400 MHz, DMSO-d6) δ 10.06 (br s, 1H), 9.44 - 9.19 (m, 1H), 8.99 (s, 1H), 8.40 (s, 1H), 8.07 (s, 1H), 7.44 (br d, J = 8.2 Hz, 3H), 7.39 - 7.10 (m, 6H), 7.08 - 7.04 (m, 1H), 6.89 - 6.77 (m, 1H), 6.24 - 6.14 (m, 1H), 5.80 - 5.73 (m, 1H), 5.72 - 5.56 (m, 1H), 4.89 (br t, J = 6.6 Hz, 1H), 4.55 (br t, J = 9.1 Hz, 1H), 4.45 - 4.40 (m, 2H), 4.29 (br s, 2H), 4.03 - 3.94 (m, 6H), 3.87 (br s, 2H), 3.80 (br s, 2H), 3.68 (br s, 1H), 3.64 - 3.49 (m, 8H), 3.43 - 3.34 (m, 1H), 3.27 - 2.97 (m, 2H), 2.45 (d, J = 2.0 Hz, 3H), 2.20 - 2.11 (m, 1H), 2.11 - 1.87 (m, 3H), 1.78 (br s, 2H), 1.49 - 1.31 (m, 6H), 0.93 (br d, J = 5.5 Hz, 9H).

[0233] Exemplary example of (2S,4R)-1-((S)-3,3-dimethyl-2-(2-(piperidine-4-ylmethoxy)acetamide)butanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide Step 1: Preparation of tert-butyl 4-[(2-ethoxy-2-oxo-ethoxy)methyl]piperidine-1-carboxylate [ka] To a solution of tert-butyl 4-(hydroxymethyl)piperidine-1-carboxylate (2.00 g, 9.29 mmol, 1.00 eq) in CH2Cl2 (50 mL), diacetoxyrhodium (1.20 g, 4.64 mmol, 0.50 eq) and ethyl 2-diazoacetate (12.00 g, 92.90 mmol, 10 mL, 10.00 eq) were added at 0°C, and the reaction mixture was stirred at 25°C for 12 hours. The solution was concentrated under reduced pressure and dried under vacuum. Purification by column chromatography with SiO2 (0-20% siRNA in petroleum ether) yielded tert-butyl 4-[(2-ethoxy-2-oxo-ethoxy)methyl]piperidine-1-carboxylate (1.50 g, 4.98 mmol, 54% yield) as a yellow oil. LC / MS(ESI)m / z:202.2[M-Boc+1] + . Step 2: Preparation of 2-[(1-tert-butoxycarbonyl-4-piperidyl)methoxyacetic acid] [ka] To a solution of tert-butyl 4-[(2-ethoxy-2-oxo-ethoxy)methyl]piperidine-1-carboxylate (1.50 g, 4.98 mmol, 1.00 eq) in THF (10 mL), CH3OH (5 mL), and H2O (5 mL), LiOH hydrate (700 mg, 14.93 mmol, 3.00 eq) was added, and the reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was acidified with dilute hydrochloric acid (pH=5), and the resulting mixture was concentrated under reduced pressure to obtain 2-[(1-tert-butoxycarbonyl-4-piperidyl)methoxy]acetic acid (1.00 g, 3.66 mmol, 74% yield) as a yellow oil. Step 3: Preparation of tert-butyl 4-[[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]methyl]piperidine-1-carboxylate [ka] CH2Cl2 (4 mL) contains 2-[(1-tert-butoxycarbonyl-4-piperidyl)methoxy]acetic acid (115 mg, 0.42 mmol, 1.00 eq) and (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (200 mg, To a solution of (0.42 mmol, 1.00 eq, hydrochloride), hydroxybenzotriazole (85 mg, 0.62 mmol, 1.50 eq), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (120 mg, 0.62 mmol, 1.50 eq), and diisopropylethylamine (270 mg, 2.08 mmol, 5.00 eq) were added, and the reaction mixture was stirred at 25°C for 12 hours. The solution was concentrated under reduced pressure and dried under vacuum. Purification by thin-layer chromatography (siRNA / CH3OH=20 / 1) yielded tert-butyl 4-[[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]methyl]piperidine-1-carboxylate (210 mg, 0.30 mmol, 72% yield) as a yellow oil. LC / MS(ESI) m / z: 700.4[M+H] + . Step 4: Preparation of (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-(4-piperidylmethoxy)acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] To a solution of tert-butyl 4-[[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]methyl]piperidine-1-carboxylate (100 mg, 0.14 mmol, 1.00 eq), HCl (4 M in dioxane, 2 mL) was added, and the reaction mixture was stirred at 25°C for 0.5 hours. The solution was concentrated under reduced pressure and dried under vacuum to obtain (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-(4-piperidylmethoxy)acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (90 mg, 0.14 mmol, 99% yield, hydrochloride) as a yellow solid.

[0234] Exemplary synthesis of tert-butyl 4-[[5-[1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2-methyl-propyl]isoxazole-3-yl]oxymethyl]piperidine-1-carboxylate Step 1: Preparation of tert-butyl 4-[[5-(1-methoxycarbonyl-2-methyl-propyl)isoxazole-3-yl]oxymethyl]piperidine-1-carboxylate [ka] To a solution of tert-butyl 4-(hydroxymethyl)piperidine-1-carboxylate (1.30 g, 6.02 mmol, 1.2 eq) and methyl 2-(3-hydroxyisoxazole-5-yl)-3-methyl-butanoate (1 g, 5.02 mmol, 1 eq) in THP (10 mL), Ph3P (1.58 g, 6.02 mmol, 1.2 eq) and diisopropyl azodicarboxylate (1.22 g, 6.02 mmol, 1.17 mL, 1.2 eq) were added, and the reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by flash silica gel chromatography (0-60% ethyl ether in petroleum ether) to obtain tert-butyl 4-[[5-(1-methoxycarbonyl-2-methyl-propyl)isoxazole-3-yl]oxymethyl]piperidine-1-carboxylate (1.8 g, 4.54 mmol, 90% yield) as a white solid. LC / MS(ESI) m / z: 297.2[M+H] + ; 1 H-NMR (400 MHz, CDCl3) δ 5.88 (s, 1H), 4.21 - 4.10 (m, 2H), 4.07 (d, J = 6.4 Hz, 2H), 3.73 (s, 3H), 3.49 (d, J = 8.7 Hz, 1H), 2.82 - 2.62 (m, 2H), 2.35 (qd, J = 7.1, 14.2 Hz, 1H), 1.96 (br d, J = 3.4 Hz, 1H), 1.77 (br d, J = 12.8 Hz, 2H), 1.46 (s, 9H), 1.29 - 1.22 (m, 2H), 1.00 (d, J = 6.7 Hz, 3H), 0.93 (d, J = 6.7 Hz, 3H). Step 2: Preparation of 2-[3-[(1-tert-butoxycarbonyl-4-piperidyl)methoxy]isoxazole-5-yl]-3-methylbutanoic acid [ka] To a solution of tert-butyl 4-[[5-(1-methoxycarbonyl-2-methyl-propyl)isoxazole-3-yl]oxymethyl]piperidine-1-carboxylate (1.8 g, 4.54 mmol, 1 eq) in THF (8 mL), CH3OH (5 mL), and H2O (3 mL), LiOH monohydrate (544 mg, 22.70 mmol, 5 eq) was added, and the reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was acidified by adding 1 M hydrochloric acid (pH=3), and the resulting precipitate was filtered to obtain crude 2-[3-[(1-tert-butoxycarbonyl-4-piperidyl)methoxy]isoxazole-5-yl]-3-methyl-butanoic acid (1.9 g) as a white solid. Step 3: Preparation of tert-butyl 4-[[5-[1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2-methyl-propyl]isoxazole-3-yl]oxymethyl]piperidine-1-carboxylate [ka] To a solution of (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (96 mg, 0.26 mmol, 1 eq, HCl salt) and 2-[3-[(1-tert-butoxycarbonyl-4-piperidyl)methoxy]isoxazole-5-yl]-3-methyl-butanoic acid (100 mg, 0.26 mmol, 1 eq), diisopropylethylamine (101 mg, 0.78 mmol, 137 μL, 3 eq) and O-(7-azabenzotriazole-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (149 mg, 0.39 mmol, 1.5 eq) were added at 0°C, and the reaction mixture was stirred at 20°C for 1 hour. The mixture was partitioned between H2O and siRNA. The organic phase was separated, washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by prep-TLC (SiO2, CH2Cl2:CH3OH=10:1) to obtain tert-butyl 4-[[5-[1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2-methyl-propyl]isoxazole-3-yl]oxymethyl]piperidine-1-carboxylate (130 mg, 0.18 mmol, 70% yield, 98% purity) as a yellow solid. LC / MS(ESI) m / z: 696.3[M+H] + This substance was separated by SFC to obtain tert-butyl 4-[[5-[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2-methyl-propyl]isoxazole-3-yl]oxymethyl]piperidine-1-carboxylate and tert-butyl 4-[[5-[(1R)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2-methyl-propyl]isoxazole-3-yl]oxymethyl]piperidine-1-carboxylate.

[0235] 1 1H NMR: tert-butyl 4-[[5-[(1R)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2-methyl-propyl]isoxazole-3-yl]oxymethyl]piperidine-1-carboxylate

[0236] 1 H-NMR (400 MHz, CDCl3) δ 8.68 (s, 1H), 7.48 (br d, J = 7.5 Hz, 1H), 7.39 (q, J = 8.3 Hz, 4H), 5.88 (s, 1H), 5.07 (t, J = 7.2 Hz, 1H), 4.67 - 4.60 (m, 2H), 4.19 - 4.02 (m, 4H), 3.80 (dd, J = 5.1, 10.5 Hz, 1H), 3.60 (dd, J = 3.7, 10.4 Hz, 1H), 3.53 - 3.48 (m, 1H), 2.78 - 2.66 (m, 2H), 2.58 - 2.47 (m, 4H), 2.46 - 2.35 (m, 1H), 1.96 (ddd, J = 5.0, 8.0, 12.8 Hz, 2H), 1.75 (br d, J = 12.1 Hz, 3H), 1.50 (d, J = 7.0 Hz, 3H), 1.46 (s, 9H), 1.28 - 1.19 (m, 2H), 1.05 (d, J = 6.6 Hz, 3H), 0.93 (d, J = 6.7 Hz, 3H).

[0237] 1 1H NMR: tert-butyl 4-[[5-[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2-methyl-propyl]isoxazole-3-yl]oxymethyl]piperidine-1-carboxylate

[0238] 1 H-NMR (400 MHz, CDCl3) δ 8.70 - 8.68 (m, 1H), 7.42 - 7.39 (m, 2H), 7.36 - 7.32 (m, 3H), 5.86 (s, 1H), 5.02 - 4.94 (m, 1H), 4.78 (dd, J = 4.3, 8.3 Hz, 1H), 4.66 (quin, J = 5.4 Hz, 1H), 4.16 - 4.09 (m, 1H), 4.08 - 4.01 (m, 2H), 3.74 - 3.68 (m, 1H), 3.60 - 3.48 (m, 2H), 2.78 - 2.62 (m, 3H), 2.52 - 2.41 (m, 1H), 2.02 - 1.93 (m, 2H), 1.75 (br d, J = 9.9 Hz, 6H), 1.46 (s, 9H), 1.38 (d, J = 7.0 Hz, 3H), 1.30 - 1.18 (m, 3H), 1.06 (d, J = 6.6 Hz, 3H), 0.94 (d, J = 6.7 Hz, 3H).

[0239] Exemplary synthesis of (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-[(5-chloro-6-fluoro-1H-indazole-4-yl)oxy]-4-(4-prop-2-enoylpiperazine-1-yl)pyrido[3,4-d]pyrimidine-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide Step 1: Preparation of 2,6-dichloro-3-fluoropyridine-4-carboxylic acid [ka] To a solution of 2,6-dichloro-3-fluoropyridine (14.2 g, 85.55 mmol, 1 eq) in THF (200 mL) cooled to -78°C, n-butyllithium (2.5 M, 41.07 mL, 1.2 eq) was added dropwise, and the reaction mixture was stirred at -78°C for 1 hour. CO2 (4.52 g, 102.66 mmol, dry ice, 1.2 eq) was added gradually, and the reaction mixture was stirred at -78°C for a further 2 hours. Saturated aqueous solution NH4Cl (30 mL), followed by 1N aqueous solution H2SO4, was added until the mixture was neutralized (pH=7). The resulting mixture was extracted with 2-methoxy-2-methylpropane (2 x 50 mL), and the combined organic extract was concentrated under vacuum. The aqueous extract was acidified by the addition of 1N aqueous solution H2SO4 (pH=1). The resulting mixture was extracted with toluene (2 x 150 mL), the combined organic extract was dried over Na2SO4, and concentrated under vacuum to obtain 2,6-dichloro-3-fluoropyridine-4-carboxylic acid (14 g, 66.67 mmol, 78% yield) as a yellow solid. 1 H-NMR (400 MHz, DMSO-d6) δ 7.85 (d, J = 4.4, 1H). Step 2: Preparation of 2,6-dichloro-3-fluoropyridine-4-carbonyl chloride [ka] To a solution of 2,6-dichloro-3-fluoropyridine-4-carboxylic acid (12 g, 57.15 mmol, 1 eq) in thionyl chloride (78.72 g, 661.68 mmol, 48.00 mL, 11.58 eq), DMF (0.02 g, 0.27 mmol, 4.79 e-3 eq) was added, and the reaction mixture was stirred at 80°C for 3 hours. The reaction mixture was concentrated under vacuum, and the resulting residue was added to toluene (50 mL). The mixture was concentrated to obtain 2,6-dichloro-3-fluoropyridine-4-carbonyl chloride (13 g, 56.91 mmol, 99% yield) as a yellow, rubbery substance. Step 3: Preparation of 2,6-dichloro-3-fluoro-N-(methylsulfanylcarbonimidoyl)pyridine-4-carboxamide [ka] To a solution of NaOH (10.24 g, 256.09 mmol, 4.5 eq) in H2O (200 mL), 2-methylisothiourea (15 g, 79.69 mmol, 1.40 eq, sulfate) was added gradually at 0°C, and the resulting mixture was stirred for 10 minutes. Then, a solution of 2,6-dichloro-3-fluoropyridine-4-carbonyl chloride (13 g, 56.91 mmol, 1 eq) in THF (150 mL) was added dropwise at 0-5°C, and the reaction mixture was stirred at 0-5°C for 30 minutes. siRNA (150 mL) and H2O (200 mL) were added, the layers were separated, and the aqueous layer was further extracted with siRNA (2 × 50 mL). The combined organic extracts were washed with H2O (2 × 80 mL), followed by brine (100 mL), dried over Na2SO3, and concentrated to obtain crude 2,6-dichloro-3-fluoro-N-(methylsulfanylcarbonimidoyl)pyridine-4-carboxamide (16 g) as a yellow solid. Step 4: Preparation of 6,8-dichloro-2-methylsulfanyl-3H-pyrido[3,4-d]pyrimidine-4-one [ka] To a solution of 2,6-dichloro-3-fluoro-N-(methylsulfanylcarbonimidoyl)pyridine-4-carboxamide (16 g, 56.71 mmol, 1 eq) in DMF (100 mL), Cs2CO3 (25.87 g, 79.40 mmol, 1.4 eq) was added, and the reaction mixture was stirred at 90°C for 5 hours. The mixture was cooled to 25°C, diluted with H2O (200 mL), and acidified by the addition of 3 M aqueous acetic acid (pH=6). The resulting precipitate was collected by filtration, the filter cake was washed with H2O (3 × 50 mL), and then dried under vacuum. The crude product obtained was purified by SiO2 column chromatography (0-20% phenyl in petroleum ether) to obtain 6,8-dichloro-2-methylsulfanyl-3H-pyrido[3,4-d]pyrimidine-4-one (2.8 g, 10.68 mmol, 19% yield) as a yellow solid. 1H-NMR (400 MHz, CDCl3) δ 13.21 (s, 1H), 7.85 (s, 1H), 1.45 (s, 9H), 2.61 (s, 3H). Step 5: Preparation of 4,6,8-trichloro-2-methylsulfanylpyrido[3,4-d]pyrimidine [ka] A mixture of 6,8-dichloro-2-methylsulfanyl-3H-pyrido[3,4-d]pyrimidine-4-one (2.5 g, 9.54 mmol, 1 eq) and POCl3 (30.94 g, 201.77 mmol, 18.75 mL, 21.15 eq) was heated at 130°C for 3 hours. The mixture was cooled to 25°C and then concentrated under vacuum. The resulting residue was dissolved in HCl (30 mL), the resulting organic mixture was washed with water (30 mL), dried over Na2SO4, and concentrated under vacuum. The resulting residue was purified by SiO2 column chromatography (0-20% HCl in petroleum ether) to obtain 4,6,8-trichloro-2-methylsulfanyl-pyrido[3,4-d]pyrimidine (750 mg, 2.67 mmol, 28% yield) as a yellow solid. Step 6: Preparation of tert-butyl 4-(6,8-dichloro-2-methylsulfanylpyrido[3,4-d]pyrimidine-4-yl)piperazine-1-carboxylate [ka] To a solution of 4,6,8-trichloro-2-methylsulfanylpyrido[3,4-d]pyrimidine (750 mg, 2.67 mmol, 1 eq) and triethylamine (541 mg, 5.35 mmol, 2 eq) in CH2Cl2 (12 mL), tert-butylpiperazine-1-carboxylate (448 mg, 2.41 mmol, 0.9 eq) was added, and the reaction mixture was stirred at 20°C for 3 hours. The mixture was concentrated under vacuum, and the resulting residue was purified by flash chromatography with SiO2 (0-10% ethyl ether in petroleum ether) to obtain tert-butyl 4-(6,8-dichloro-2-methylsulfanylpyrido[3,4-d]pyrimidine-4-yl)piperazine-1-carboxylate (680 mg, 1.58 mmol, 59% yield) as a yellow, rubbery substance. Step 7: Preparation of 3-bromo-5-fluoro-2-methylaniline [ka] To a mixture of 1-bromo-5-fluoro-2-methyl-3-nitrobenzene (15 g, 64.10 mmol, 1 eq) and NH4Cl (17.14 g, 320.48 mmol, 5 eq) in 30 mL of H2O and 150 mL of ethanol at 80°C, Fe (17.90 g, 320.48 mmol, 5 eq) was gradually added, and the reaction mixture was stirred at 80°C for 14 hours. The mixture was filtered, and the filtrate was extracted with HCl (3 x 20 mL). The combined organic extract was washed with brine (3 x 20 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The resulting residue was purified by column chromatography in SiO2 (0-100% HCl in petroleum ether) to obtain the desired product (4.593 g). An additional (4.684 g) of the substance was obtained by further purification of the impure fraction by semi-preparative reverse-phase HPLC (30-60% CH3CN in water (0.1% TFA)). Compound 3-bromo-5-fluoro-2-methylaniline (9.28 g, 45.34 mmol, 71% yield, 99% purity) was obtained as a brown oil. 1H-NMR (400 MHz, CDCl3) δ 7.75 (dd, J = 2.4, 10.0 Hz, 1H), 4.48 (s, 2H), 2.20 (s, 3H). Step 8: Preparation of 3-bromo-4-chloro-5-fluoro-2-methylaniline [ka] To a solution of 3-bromo-5-fluoro-2-methylaniline (9.28 g, 45.47 mmol, 1 eq) in isopropanol (50 mL), 1-chloropyrrolidine-2,5-dione (6.68 g, 50.01 mmol, 1.1 eq) was added, and the reaction mixture was stirred at 80°C for 2 hours. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by flash chromatography with SiO2 (10-30% ethyl phosphate in petroleum ether), followed by preparative reverse-phase HPLC (45-75% CH3CN in water (0.05% HCl)) to obtain 3-bromo-4-chloro-5-fluoro-2-methylaniline (4.87 g, 20.36 mmol, 45% yield, 99% purity) as a white solid. 1 H-NMR (400 MHz, DMSO-d6) δ 6.62 (d, J = 11.6 Hz, 1H), 2.18 (m, 3H). Step 9: Preparation of 4-bromo-5-chloro-6-fluoro-1H-indazole [ka] To a solution of 3-bromo-4-chloro-5-fluoro-2-methylaniline (4.87 g, 20.42 mmol, 1 eq) in acetic acid (40 mL), NaNO2 (1.80 g, 26.14 mmol, 1.28 eq) was added, and the reaction mixture was stirred at 25°C for 7 hours, followed by 40°C for 14 hours. The mixture was diluted with water (200 mL) and extracted with siRNA (3 x 50 mL). The combined organic extract was washed with brine (3 x 30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by column chromatography in SiO2 (10-100% CH2Cl2 in petroleum ether). The product was washed with CH2Cl2 (100 mL) to obtain pure 4-bromo-5-chloro-6-fluoro-1H-indazole (751 mg, 3.01 mmol, 14.74% yield) and an additional crude product (1.42 g). 1 H-NMR (400 MHz, DMSO-d6) δ 13.68 (s, 1H), 8.10 (s, 1H), 7.70 (d, J = 9.2 Hz, 1H). Step 10: Preparation of 4-bromo-5-chloro-6-fluoro-1-tetrahydropyran-2-yl-indazole [ka] To a solution of 4-bromo-5-chloro-6-fluoro-1H-indazole (1.2 g, 4.81 mmol, 1 eq) in CH2Cl2 (50 mL), p-toluenesulfonic acid (92 mg, 0.48 mmol, 0.1 eq) and 3,4-dihydro-2H-pyran (809 mg, 9.62 mmol, 2 eq) were added, and the reaction mixture was stirred at 20°C for 0.5 hours. The mixture was diluted with H2O (30 mL) and extracted with Âx (3 x 20 mL). The combined organic extract was washed with brine (3 x 30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by column chromatography in SiO2 (3-10% siRNA in petroleum ether) to obtain 4-bromo-5-chloro-6-fluoro-1-tetrahydropyran-2-yl-indazole (1.26 g, 3.78 mmol, 78% yield) as a yellow oil. 1H-NMR (400 MHz, CDCl3) δ 7.86 (s, 1H), 7.27 (d, J = 9.2 Hz, 1H), 5.53 (dd, J = 2.4, 9.2 Hz, 1H), 3.87 - 3.85 (m, 1H), 3.65 - 3.60 (m, 1H), 2.36 - 2.33 (m, 1H), 2.04 - 2.00 (m, 2H), 1.65 - 1.57 (m, 3H). Step 11: Preparation of 5-chloro-6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-4-ol [ka] To 4-bromo-5-chloro-6-fluoro-1-tetrahydropyran-2-yl-indazole (2.24 g, 6.71 mmol, 1 eq) in dioxane (30 mL), tris(dibenzylideneacetone)dipalladium (0) (307 mg, 0.34 mmol, 0.05 eq), ditert-butyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane (285 mg, 0.67 mmol, 0.1 eq), followed by KOH (1.13 g, 20.14 mmol, 3 eq) in H2O (8 mL). The reaction mixture was degassed, charged with N2 (3X), and then stirred at 90°C for 16 hours under an N2 atmosphere. The resulting residue was partitioned between petroleum ether (50 mL) and water. The aqueous layer was extracted with petroleum ether (3X 30 mL), and the combined organic extract was discarded. The aqueous phase was acidified by adding 1N aqueous HCl (pH=3), and then extracted with siRNA (3 x 30 mL). The combined organic extract was washed with water (50 mL), followed by brine (100 mL), dried over Na₂SO₄, and concentrated until dry. The resulting residue was purified by flash chromatography in SiO₂ (0-20% siRNA in petroleum ether) to obtain 5-chloro-6-fluoro-1-tetrahydropyran-2-ylindazole-4-ol (1.44 g, 5.31 mmol, 79% yield) as a yellow solid. 1H-NMR (400 MHz, CDCl3) δ 8.08 (s, 1H), 7.07 (d, J = 0.8 Hz, 1H), 6.31 (s, 1H), 5.60 (dd, J = 2.4, 9.2 Hz, 1H), 4.14 - 4.00 (m, 1H), 3.74 - 3.73 (m, 1H), 2.51 - 2.48 (m, 1H), 2.14 - 2.06 (m, 2H), 1.77 - 1.67 (m, 3H). Step 12: Preparation of tert-butyl 4-[6-chloro-8-(5-chloro-6-fluoro-1-tetrahydropyran-2-yl-indazole-4-yl)oxy-2-methylsulfanyl-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate [ka] A mixture of 5-chloro-6-fluoro-1-tetrahydropyran-2-yl-indazole-4-ol (300 mg, 1.11 mmol, 1 eq) and tert-butyl 4-(6,8-dichloro-2-methylsulfanylpyrido[3,4-d]pyrimidine-4-yl)piperazine-1-carboxylate (501 mg, 1.16 mmol, 1.05 eq) in dried DMA (6 mL) was mixed with Cs2CO3 (541.65 mg, 1.66 mmol, 1.5 eq), and the reaction mixture was stirred at 85°C for 5 hours. The mixture was cooled to 25°C and then concentrated under vacuum. The remaining residue was dissolved in Â(40 mL), the resulting organic mixture was washed with water (30 mL), dried over Na2SO4, and concentrated under vacuum. The obtained residue was purified by column chromatography in SiO2 (0-20% ethyl ether in petroleum ether) to obtain tert-butyl 4-[6-chloro-8-(5-chloro-6-fluoro-1-tetrahydropyran-2-yl-indazole-4-yl)oxy-2-methylsulfanyl-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (420 mg, 0.63 mmol, 57% yield) as a yellow solid. 1H-NMR (400 MHz, CDCl3) δ 7.87 (s, 1H), 7.34 (dd, J = 0.8, 8.4 Hz, 1H), 7.32 (s, 1H), 4.08 - 4.04 (m, 1H), 3.84 - 3.82 (m, 4H), 3.79 - 3.76 (m, 1H), 3.69 - 3.66 (m, 4H), 2.60 (s, 3H), 2.70 - 2.48 (m, 1H), 2.17 - 2.13 (m, 1H), 1.79 -1.71 (m, 3H), 1.52 (s, 9H). Step 13: Preparation of tert-butyl 4-(6-chloro-8-((5-chloro-6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-4-yl)oxy)-2-(methylsulfonyl)pyrido[3,4-d]pyrimidine-4-yl)piperazine-1-carboxylate [ka] To a solution of tert-butyl 4-[6-chloro-8-(5-chloro-6-fluoro-1-tetrahydropyran-2-yl-indazole-4-yl)oxy-2-methylsulfanylpyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (320 mg, 0.48 mmol, 1 eq) in CH2Cl2 (8 mL), 3-chloroperbenzoic acid (196 mg, 0.96 mmol, 85% purity, 2 eq) was added, and the reaction mixture was stirred at 20°C for 16 hours. The mixture was diluted with CH2Cl2 (30 mL), the resulting organic mixture was washed with water (30 mL), dried over Na2SO4, and concentrated under vacuum. The obtained residue was purified by prep-TLC (80% ethyl ether in petroleum ether) to obtain tert-butyl 4-[6-chloro-8-(5-chloro-6-fluoro-1-tetrahydropyran-2-yl-indazole-4-yl)oxy-2-methylsulfonyl-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (230 mg, 0.33 bmmol, 68% yield) as a yellow solid. LC / MS (ESI) m / z: 696.1 [M+H] + . Step 13: Preparation of tert-butyl 4-[6-chloro-8-(5-chloro-6-fluoro-1-tetrahydropyran-2-yl-indazole-4-yl)oxy-2-[(1R)-2,2-dimethoxy-1-methylethoxy]pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate [ka] To a solution of tert-butyl 4-[6-chloro-8-(5-chloro-6-fluoro-1-tetrahydropyran-2-yl-indazole-4-yl)oxy-2-methylsulfonylpyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (100 mg, 0.14 mmol, 1 eq) and (2R)-1,1-dimethoxypropan-2-ol (35 mg, 0.29 mmol, 2 eq), LiHMDS (1 M, 0.22, 1.5 eq) was added in 3 mL of THF at 0°C, and the reaction mixture was stirred at 0-5°C for 30 minutes. Saturated aqueous solution NH4Cl (15 mL) and siRNA (20 mL) were added, the organic layer was separated, dried over Na2SO4, and concentrated under vacuum. The obtained residue was purified by prep-TLC (33% ethyl ether in petroleum ether) to obtain tert-butyl 4-[6-chloro-8-(5-chloro-6-fluoro-1-tetrahydropyran-2-yl-indazole-4-yl)oxy-2-[(1R)-2,2-dimethoxy-1-methylethoxy]pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (70 mg, 0.095 mmol, 66% yield) as a yellow solid. Step 14: Preparation of (R)-2-((6-chloro-8-((5-chloro-6-fluoro-1H-indazole-4-yl)oxy)-4-(piperazine-1-yl)pyrido[3,4-d]pyrimidine-2-yl)oxy)propanal [ka] To a solution of tert-butyl 4-[6-chloro-8-(5-chloro-6-fluoro-1-tetrahydropyran-2-yl-indazole-4-yl)oxy-2-[(1R)-2,2-dimethoxy-1-methylethoxy]pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (70 mg, 0.095 mmol, 1 eq) in dioxane (3 mL), aqueous HCl (12 M, 0.5 mL, 63.14 eq) was added, and the reaction mixture was stirred at 15°C for 1 hour. The mixture was concentrated under vacuum to obtain (R)-2-((6-chloro-8-((5-chloro-6-fluoro-1H-indazole-4-yl)oxy)-4-(piperazin-1-yl)pyrido[3,4-d]pyrimidine-2-yl)oxy)propanal (61 mg, 0.095 mmol, 99% yield, hydrochloride) as a yellow solid. LC / MS (ESI) m / z: 505.9 [M+H] + . Step 15: Preparation of (2R)-2-[6-chloro-8-[(5-chloro-6-fluoro-1H-indazole-4-yl)oxy]-4-(4-prop-2-enoylpiperazine-1-yl)pyrido[3,4-d]pyrimidine-2-yl]oxypropanal [ka] A mixture of (2R)-2-[6-chloro-8-[(5-chloro-6-fluoro-1H-indazole-4-yl)oxy]-4-piperazine-1-ylpyrido[3,4-d]pyrimidine-2-yl]oxypropanal (61 mg, 0.11 mmol, 1 eq, hydrochloride) and NaHCO3 (6.48 g, 3 mL) in THF (0.9 mL) was mixed with a solution of prop-2-enoyl chloride (10 mg, 0.11 mmol, 9.16 μL, 1 eq) in THF (0.9 mL), and the reaction mixture was stirred at 15°C for 20 minutes. ELISA (20 mL) and water (20 mL) were added, the organic layer was separated, dried over Na2SO4, and concentrated under vacuum. The obtained residue was purified by prep-TLC (10% CH3OH in CH2Cl2) to obtain (2R)-2-[6-chloro-8-[(5-chloro-6-fluoro-1H-indazole-4-yl)oxy]-4-(4-prop-2-enoylpiperazin-1-yl)pyrido[3,4-d]pyrimidine-2-yl]oxypropanal (37 mg, 0.066 mmol, 59% yield) as a yellow solid. LC / MS (ESI) m / z: 560.1 [M+H] + . Step 16: Preparation of (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-[(5-chloro-6-fluoro-1H-indazole-4-yl)oxy]-4-(4-prop-2-enoylpiperazin-1-yl)pyrido[3,4-d]pyrimidine-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] To a solution of (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-(4-piperidyloxy)ethoxy]acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (50 mg, 0.075 mmol, 1.2 eq, hydrochloride) in CH3OH (1 mL), NaOAc (15 mg, 187.38 µl, 3 eq) was added, and the resulting mixture was stirred at 20°C for 20 minutes. Next, a solution of (2R)-2-[6-chloro-8-[(5-chloro-6-fluoro-1H-indazole-4-yl)oxy]-4-(4-prop-2-enoylpiperazin-1-yl)pyrido[3,4-d]pyrimidine-2-yl]oxypropanal (35 mg, 0.062 mmol, 1 eq) was added to CH2Cl2 (1 mL), and the resulting mixture was cooled to 0°C. NaBH4CN (8 mg, 0.12 mmol, 2 eq) was added, and the reaction mixture was stirred at 0-15°C for 48 hours. Water (5 mL) was added, and the resulting mixture was extracted with CH2Cl2 (5 x 3 mL). The combined organic extract was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The obtained residue was purified by prep-TLC (CH2Cl2:CH3OH=9:1) of SiO2 and freeze-dried to (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[6-chloro-8-[(5-chloro-6-fluoro-1H-indazole-4-yl)oxy]-4-(4-prop-2-enoylpiperazine-1-yl)pyrido[3,4-d [Pyrimidine-2-yl]oxypropyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (10.3 mg, 0.008 mmol, 31% yield, 90% purity) was obtained as a white solid. LC / MS (ESI) m / z: 1175.3[M+H]+.1H-NMR (400 MHz, DMSO-d6) δ 13.62 - 13.53 (m, 1H), 8.98 (s, 1H), 8.45 - 8.37 (m, 1H), 7.77 (s, 1H), 7.70 - 7.57 (m, 2H), 7.44 - 7.40 (m, 2H), 7.35 (br d, J = 8.4 Hz, 2H), 6.86 - 6.76 (m, 1H), 6.17 (dd, J = 2.3, 17.0 Hz, 1H), 5.77 - 5.70 (m, 1H), 5.41 - 5.20 (m, 1H), 5.16 - 5.09 (m, 1H), 4.94 - 4.83 (m, 1H), 4.57 - 4.49 (m, 1H), 4.46 - 4.38 (m, 1H), 4.31 - 4.24 (m, 1H), 3.94 (br s, 6H), 3.85 - 3.73 (m, 4H), 3.62 - 3.54 (m, 4H), 3.53 - 3.47 (m, 2H), 2.84 - 2.70 (m, 2H), 2.45 (s, 4H), 2.22 - 2.09 (m, 3H), 2.07 - 1.93 (m, 3H), 1.81 - 1.71 (m, 3H), 1.39 - 1.32 (m, 5H), 1.30 - 1.25 (m, 3H), 0.91 (s, 9H).

[0240] Exemplary synthesis of (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoylpiperazine-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide Step 1: Preparation of tert-butyl 4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-7-carboxylate [ka] 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 (2.50 g, 4.74 mmol, 1.00 eq) and (2R)-1,1-dimethoxypropan-2-ol (1.70 g, 14.23 mmol, 3.00 eq), Cs2CO3 (4.80 g, 14.23 mmol, 3.00 eq) and [2-(2-aminophenyl)phenyl]-methylsulfonyloxypalladium;dicyclohexyl-[2-(2,6-diisopropoxyphenyl)phenyl]phosphane (278 mg, 0.33 mmol, 0.07 eq) were added to dioxane (15 mL), and the reaction mixture was stirred at 90°C for 6 hours. The solution was concentrated, and the resulting substance was purified by column chromatography in SiO2 (10-100% SiO2 in petroleum ether) to obtain tert-butyl 4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-7-carboxylate (1.00 g, 1.64 mmol, 35% yield) as a yellow solid. LC / MS(ESI) m / z: 611.2[M+H] + . Step 2: Preparation of benzyl(2S)-2-(cyanomethyl)-4-[2-[(1R)-2,2-dimethoxy-1-methylethoxy]-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate [ka] To a solution of tert-butyl 4-[(3S)-4-benzyloxycarbonyl-3-(cyanomethyl)piperazin-1-yl]-2-[(1R)-2,2-dimethoxy-1-methylethoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-7-carboxylate (1.00 g, 1.64 mmol, 1.00 eq) in CH2Cl2 (10 mL), TFA (1.50 g, 13.51 mmol, 1 mL, 8.25 eq) was added, and the reaction mixture was stirred at 25°C for 2 hours. The mixture was neutralized by the addition of NaHCO3 (pH=7) and then extracted with ELISA (2 x 100 mL). The combined organic extracts were concentrated under reduced pressure to obtain benzyl(2S)-2-(cyanomethyl)-4-[2-[(1R)-2,2-dimethoxy-1-methylethoxy]-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (800 mg, 1.57 mmol, 96% yield) as a yellow solid. LC / MS(ESI) m / z: 511.5[M+H] + . Step 3: Preparation of benzyl(2S)-2-(cyanomethyl)-4-[2-[(1R)-2,2-dimethoxy-1-methylethoxy]-7-[3-(methoxymethoxy)-1-naphthyl]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate [ka] A solution of benzyl(2S)-2-(cyanomethyl)-4-[2-[(1R)-2,2-dimethoxy-1-methylethoxy]-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (800 mg, 1.57 mmol, 1.00 eq) and 1-bromo-3-(methoxymethoxy)naphthalene (465 mg, 1.72 mmol, 1.10 eq) in dioxane (10 mL) [2-(2-aminophenyl)phenyl]-methylsulfonyloxypalladium;dicyclohexyl-[2-(2,6-diisopropoxyphenyl)phenyl]phosphan (90 mg, 0.11 mmol, 0.07 eq), Cs2CO3 (1.50 g, 4.70 mmol, 3.00 eq), and Pd(OAc)2 (20 mg, 0.08 mmol, 0.05 eq) were added, and the reaction mixture was stirred at 90°C for 6 hours under an N2 atmosphere. The mixture was concentrated, and the resulting substance was purified by column chromatography in SiO2 (10-100% SiO2 in petroleum ether) to obtain benzyl(2S)-2-(cyanomethyl)-4-[2-[(1R)-2,2-dimethoxy-1-methylethoxy]-7-[3-(methoxymethoxy)-1-naphthyl]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (480 mg, 0.69 mmol, 44% yield) as a yellow solid. LC / MS(ESI) m / z: 697.2[M+H] + . Step 4: Preparation of 2-[(2S)-4-[2-[(1R)-2,2-dimethoxy-1-methylethoxy]-7-[3-(methoxymethoxy)-1-naphthyl]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-2-yl]acetonitrile [ka] CH 3To a solution of benzyl(2S)-2-(cyanomethyl)-4-[2-[(1R)-2,2-dimethoxy-1-methylethoxy]-7-[3-(methoxymethoxy)-1-naphthyl]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (460 mg, 0.66 mmol, 1.00 eq) in OH (15 mL), Pd / C carbon (100 mg) and NH4OH (0.3 mL, 37% purity) were added, and the reaction mixture was stirred at 25°C for 1 hour under an H2 atmosphere. The solution was concentrated to obtain 2-[(2S)-4-[2-[(1R)-2,2-dimethoxy-1-methylethoxy]-7-[3-(methoxymethoxy)-1-naphthyl]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazin-2-yl]acetonitrile (360 mg, 0.64 mmol, 96% yield) as a yellow solid. LC / MS(ESI) m / z: 563.4[M+H] + . Step 5: Preparation of 2-[(2S)-4-[7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxoethoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazin-2-yl]acetonitrile [ka] To a solution of 2-[(2S)-4-[2-[(1R)-2,2-dimethoxy-1-methylethoxy]-7-[3-(methoxymethoxy)-1-naphthyl]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazin-2-yl]acetonitrile (210 mg, 0.37 mmol, 1.00 eq) in CH2Cl2 (3 mL), HCl (4N in dioxane, 1 mL) was added, and the reaction mixture was stirred at 25°C for 30 minutes. The solution was concentrated under reduced pressure and basicized by the addition of NaHCO3 (pH=8). The resulting basic mixture was extracted with ELISA (2 x 50 mL), and the combined organic extract was concentrated to obtain 2-[(2S)-4-[7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxo-ethoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazin-2-yl]acetonitrile (170 mg, 0.36 mmol, 96% yield) as a yellow solid. Step 6: Preparation of tert-butyl(2S)-2-(cyanomethyl)-4-[7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxoethoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate [ka] To a solution of 2-[(2S)-4-[7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxoethoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazin-2-yl]acetonitrile (170 mg, 0.36 mmol, 1.00 eq) in THF (3 mL) and H2O (1 mL), di-tert-butyl dicarbonate (88 mg, 0.36 mmol, 1.10 eq) and NaHCO3 (90 mg, 1.08 mmol, 3.00 eq) were added, and the reaction mixture was stirred at 25°C for 3 hours. The solution was concentrated, and the remaining substance was purified by thin-layer chromatography (CH2Cl2 / CH3OH=10 / 1) to obtain tert-butyl(2S)-2-(cyanomethyl)-4-[7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxoethoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (90 mg, 0.16 mmol, 44% yield) as a yellow solid. LC / MS(ESI) m / z: 573.3[M+H] + . Step 7: Preparation of tert-butyl(2S)-2-(cyanomethyl)-4-[2-[(1R)-2-[4-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-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)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate [ka] tert-butyl(2S)-2-(cyanomethyl)-4-[7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxo-ethoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (45 mg, 0.08 mmol, 1.05 eq) and (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-[2-(4-piperidyloxy)ethoxy] in CH2Cl2 (1 mL) and CH3OH (1 mL). To a solution of acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (50 mg, 0.07 mmol, 1.00 eq, hydrochloride), NaOAc (20 mg, 0.23 mmol, 3.00 eq), borane;2-methylpyridine (40 mg, 0.38 mmol, 5.00 eq), and acetic acid (15 mg, 0.23 mmol, 3.00 eq) were added, and the reaction mixture was stirred at 40°C for 12 hours. The solution is concentrated under reduced pressure, and the remaining substance is purified by thin-layer chromatography (CH2Cl2 / CH3OH=10 / 1) to obtain tert-butyl(2S)-2-(cyanomethyl)-4-[2-[(1R)-2-[4-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl] Pyrrolidine-1-carbonyl]-2,2-dimethylpropyl]amino]-2-oxoethoxy]ethoxy]-1-piperidyl]-1-methylethoxy]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (40 mg, 0.03 mmol, 45% yield) was obtained as a yellow solid. LC / MS (ESI) m / z: 1186.5 [M+H] + . Step 8: Preparation of (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)piperazin-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] tert-butyl(2S)-2-(cyanomethyl)-4-[2-[(1R)-2-[4-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl]amino]-2-oxy To a solution of so-ethoxy]ethoxy]-1-piperidyl]-1-methylethoxy]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (30 mg, 0.03 mmol, 1.00 eq), HCl (4N in dioxane, 0.5 mL) was added, and the reaction mixture was stirred at 25°C for 0.5 hours. The solution was concentrated under reduced pressure, the remaining substance was incorporated into HCl and CH3CN, and the solution was concentrated multiple times under vacuum to obtain (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)piperazin-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (28 mg, 0.02 mmol, 96% yield, dihydrochloride) as a yellow solid. LC / MS(ESI)m / z:1086.2[M+H] + . Step 9: Preparation of (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoylpiperazine-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)piperazin-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4 in CH2Cl2 (15mL) To a solution of -hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (28 mg, 0.02 mmol, 1.00 eq, 2 hydrochloride), 2,6-dimethylpyridine (20 mg, 0.19 mmol, 8.00 eq) and prop-2-enoyl chloride (2 mg, 0.02 mmol, 1.00 eq) were added, and the reaction mixture was stirred at -75°C for 0.5 hours. The solution was concentrated under reduced pressure, and the remaining substance was subjected to prep-HPLC [water (10 mM)]. Purified with 30-60% CH3CN in NH4HCO3, (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoylpiperazin-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (4.0 mg, 14% yield, 97% purity) was obtained as a white solid. LC / MS(ESI)m / z:1140.4[M+H] + . 1H-NMR (400 MHz, CD3OD) δ 8.85 (s, 1H), 8.06 (d, J = 8.8 Hz, 1H), 7.62 (d, J = 8.0 Hz, 1H), 7.44 - 7.32 (m, 5H), 7.29 - 7.19 (m, 1H), 6.99 - 6.66 (m, 3H), 6.29 (d, J = 16.8 Hz, 1H), 5.83 (d, J = 11.2 Hz, 1H), 5.42 (d, J = 2.8 Hz, 1H), 5.18 - 5.03 (m, 1H), 5.01 - 4.95 (m, 1H), 4.82 - 4.48 (m, 4H), 4.47 - 4.29 (m, 1H), 4.21 - 3.97 (m, 6H), 3.89 - 3.35 (m, 9H), 3.12 - 2.81 (m, 6H), 2.73 (dd, J = 7.6, 13.2 Hz, 1H), 2.52 (dd, J = 2.8, 13.6 Hz, 1H), 2.48 - 2.40 (m, 3H), 2.39 - 2.25 (m, 2H), 2.19 (dd, J = 7.6, 12.8 Hz, 1H), 2.03 - 1.79 (m, 3H), 1.64 - 1.53 (m, 2H), 1.47 (d, J = 7.2 Hz, 3H), 1.33 (d, J = 6.0 Hz, 3H), 1.02 (s, 9H).

[0241] The following compound can be prepared by a method similar to that of 2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoylpiperazine-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide. 1. Exemplary compound (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoylpiperazine-1-yl]-7-(1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] (Formate, white solid). LC / MS (ESI) m / z: 1124.4 [M+H] + . 1 H-NMR (400 MHz, CD3OD) δ 8.88 (s, 1H), 8.54 (s, 1H), 8.28 - 8.21 (m, 1H), 7.91 - 7.84 (m, 1H), 7.62 (d, J=8.0 Hz, 1H), 7.54 - 7.47 (m, 2H),7.46 - 7.35(m, 5H), 7.21 (d, J=7.2 Hz, 1H), 6.81 (s, 1H), 6.31 (d, J=16.4 Hz, 1H), 5.85 (d, J=10.0 Hz, 1H), 5.50 (s, 1H), 5.19 - 4.93 (m, 2H), 4.86 - 4.77 (m, 1H), 4.65 - 4.54 (m, 2H), 4.48 - 4.35 (m, 1H), 4.29 - 4.01 (m, 7H), 3.88 - 3.80 (m, 1H), 3.79 - 3.59 (m, 6H), 3.58 - 3.36 (m, 3H), 3.29 - 2.59 (m, 12H), 2.51 - 2.43 (m, 3H), 2.26 - 2.17 (m, 1H), 2.06 - 1.91 (m, 3H), 1.74 (s, 2H), 1.49 (d, J=7.2 Hz, 3H), 1.39 (d, J=6 Hz, 3H), 1.05 (s, 9H). 2. Exemplary compound (2S,4R)-1-[(2S)-2-[[2-[2-[[1-[2-[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoylpiperazine-1-yl]-7-(8-methyl-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]ethyl]-4-piperidyl]oxy]ethoxy]acetyl]amino]-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] (Formate, white solid). LC / MS (ESI) m / z: 1123.6 [M+H] + . 1 H-NMR (400MHz, CD3OD) δ 8.94 - 8.76 (m, 1H), 8.52 (d, J = 1.6 Hz, 1H), 7.66 (dd, J = 7.6, 17.2 Hz, 2H), 7.50 - 7.15 (m, 8H), 6.81 (d, J = 2.4 Hz, 1H), 6.29 (d, J = 16.0 Hz, 1H), 5.83 (d, J = 11.2 Hz, 1H), 5.16 - 4.94 (m, 2H), 4.79 (d, J = 7.6 Hz, 1H), 4.65 - 4.64 (m, 1H), 4.64 - 4.47 (m, 3H), 4.49 - 4.27 (m, 1H), 4.26 - 3.98 (m, 5H), 3.90 - 3.41 (m, 11H), 3.28 - 2.94 (m, 9H), 2.90 (s, 4H), 2.84 - 2.59 (m, 3H), 2.46 (s, 3H), 2.38 - 2.14 (m, 1H), 2.07 - 1.69 (m, 5H), 1.61 - 1.39 (m, 3H), 1.03 (s, 9H).

[0242] Exemplary synthesis of (2S,4R)-1-[(2S)-2-[[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoylpiperazine-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide Step 1: Preparation of tert-butyl(2S)-2-(cyanomethyl)-4-[2-[(1R)-2-[4-[[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]methyl]-1-piperidyl]-1-methyl-ethoxy]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate [ka] (2S,4R)-1-[(2S)-3,3-dimethyl-2-[[2-(4-piperidylmethoxy)acetyl]amino]butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (45 mg, 0.07 mmol, 1.01 eq, hydrochloric acid) and tert-butyl(2S)-2-(shear) in CH2Cl2 (0.7 mL) and CH3OH (0.7 mL). To a solution of (40 mg, 0.07 mmol, 1.00 eq) of (nomethyl)-4-[7-(3-hydroxy-1-naphthyl)-2-[(1R)-1-methyl-2-oxoethoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (40 mg, 0.07 mmol, 1.00 eq), NaOAc (60 mg, 0.70 mmol, 10.00 eq) and acetic acid (cat.) were added at 25°C, and the resulting mixture was stirred at 25°C for 2 hours. 2-methylpyridineborane (40 mg, 0.37 mmol, 5.35 eq) was added at 0°C, and the reaction mixture was stirred at 40°C for 8 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by prep-thin-layer chromatography (CH2Cl2:CH3OH=10:1) to tert-butyl(2S)-2-(cyanomethyl)-4-[2-[(1R)-2-[4-[[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyllo [Zin-1-carbonyl]-2,2-dimethylpropyl]amino]-2-oxoethoxy]methyl]-1-piperidyl]-1-methylethoxy]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (50 mg, 0.04 mmol, 61% yield) was obtained as a brown solid. LC / MS (ESI) m / z: 1156.9 [M+H] + . Step 2: Preparation of (2S,4R)-1-[(2S)-2-[[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)piperazin-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] tert-butyl(2S)-2-(cyanomethyl)-4-[2-[(1R)-2-[4-[[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethylpropyl]amino]-2-oxo-ethoxy To a solution of [methyl]-1-piperidyl]-1-methyl-ethoxy]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-4-yl]piperazine-1-carboxylate (50 mg, 0.04 mmol, 1.00 eq), TFA (13.51 mmol, 1 mL, 312.38 eq) was added, and the reaction mixture was stirred at 25°C for 10 minutes. The mixture was concentrated under reduced pressure to obtain (2S,4R)-1-[(2S)-2-[[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)piperazin-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (50 mg, 0.04 mmol, 90% yield, 2 trifluoroacetates) as a red oil. LC / MS (ESI) m / z: 1056.7[M+H] + . Step 3: Preparation of (2S,4R)-1-[(2S)-2-[[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoylpiperazine-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] (2S,4R)-1-[(2S)-2-[[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)piperazin-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4 in CH2Cl2 (4 mL) at -78°C To a solution of -hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (50 mg, 0.04 mmol, 1.00 eq, 2-trifluoroacetate), 2,6-lutidine (0.40 mmol, 10.00 eq) and prop-2-enoyl chloride (4 mg, 0.04 mmol, 1.00 eq) were added, and the reaction mixture was stirred at -78°C for 30 minutes. Water was then added at -78°C, and the resulting mixture was concentrated under reduced pressure. The obtained residue was subjected to semi-preparative reverse-phase HPLC (water (0.01M)). (2S,4R)-1-[(2S)-2-[[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)-4-prop-2-enoylpiperazin-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (7.5 mg, 0.006 mmol, 16% yield, 96% purity) was obtained as a white solid after lyophilization. LC / MS(ESI)m / z:1110.6[M+H] + . 1H-NMR (400 MHz, CD3OD) δ 8.86 (s, 1H), 8.06 (d, J = 8.4 Hz, 1H), 7.61 (d, J = 8.0 Hz, 1H), 7.46 - 7.31 (m, 5H), 7.29 - 7.20 (m, 1H), 6.94 - 6.71 (m, 3H), 6.29 (d, J = 16.4 Hz, 1H), 5.83 (d, J = 10.0 Hz, 1H), 5.44 (s, 1H), 5.16 - 4.93 (m, 2H), 4.72 - 4.32 (m, 5H), 4.25 - 4.02 (m, 5H), 4.01 - 3.90 (m, 2H), 3.86 - 3.79 (m, 1H), 3.77 - 3.69 (m, 1H), 3.63 (s, 1H), 3.38 (d, J = 6.0 Hz, 3H), 3.13 - 2.85 (m, 6H), 2.75 (dd, J = 8.4, 13.6 Hz, 1H), 2.59 - 2.50 (m, 1H), 2.46 (s, 3H), 2.25 - 2.09 (m, 3H), 1.96 (dd, J = 4.0, 9.2 Hz, 1H), 1.81 - 1.59 (m, 3H), 1.58 - 1.43 (m, 3H), 1.41 - 1.10 (m, 6H), 1.02 (s, 9H).

[0243] Exemplary synthesis of (2S,4R)-1-[(2S)-2-[[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)-4-(2-fluoroprop-2-enoyl)piperazine-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] (2S,4R)-1-[(2S)-2-[[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)piperazin-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8-dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4 To a solution of -(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (100 mg, 0.09 mmol, 1.00 eq, hydrochloride) and 2-fluoroprop-2-enoic acid (8 mg, 0.09 mmol, 1.00 eq), HATU (55 mg, 0.14 mmol, 1.50 eq) and diisopropylethylamine (25 mg, 0.18 mmol, 2.00 eq) were added, and the reaction mixture was stirred at 25°C for 1 hour. The solution is concentrated under reduced pressure, and the remaining substance is purified by prep-HPLC (25-55% CH3CN in water (0.225% formic acid)) to obtain (2S,4R)-1-[(2S)-2-[[2-[[1-[(2R)-2-[[4-[(3S)-3-(cyanomethyl)-4-(2-fluoroprop-2-enoyl)piperazin-1-yl]-7-(3-hydroxy-1-naphthyl)-6,8- Dihydro-5H-pyrido[3,4-d]pyrimidine-2-yl]oxy]propyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (5.6 mg, 5% yield, 99% purity) was obtained as a white solid. LC / MS (ESI) m / z: 1129.6[M+H] + . 1H-NMR (400 MHz, CD3OD) δ 8.79 - 8.74 (m, 1H), 7.97 (d, J = 9.2 Hz, 1H), 7.52 (d, J = 7.6 Hz, 1H), 7.36 - 7.22 (m, 7H), 7.16 (s, 1H), 6.77 (d, J = 1.6 Hz, 1H), 6.69 (d, J = 2.4 Hz, 1H), 5.42 - 5.27 (m, 1H), 5.26 - 5.05 (m, 2H), 4.92 - 4.88 (m, 1H), 4.57 (s, 1H), 4.46 (s, 2H), 4.38 - 4.22 (m, 1H), 4.16 - 4.00 (m, 5H), 3.93 - 3.79 (m, 3H), 3.77 - 3.38 (m, 4H), 3.30 (d, J = 6.4 Hz, 4H), 3.09 - 3.01 (m, 4H), 3.00 - 2.89 (m, 2H), 2.67 - 2.51 (m, 1H), 2.36 (s, 3H), 2.32 - 2.00 (m, 4H), 1.84 (dd, J = 4.0, 12.8 Hz, 1H), 1.76 - 1.64 (m, 3H), 1.37 (d, J = 7.2 Hz, 3H), 1.26 (d, J = 6.4 Hz, 5H), 0.93 - 0.91 (m, 9H).

[0244] Exemplary synthesis of (2S,4R)-1-[(2S)-2-[[2-[[1-[3-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide Step 1: Preparation of ethyl 2-(4-piperidyloxy)acetate [ka] A solution of tert-butyl 4-(2-ethoxy-2-oxo-ethoxy)piperidine-1-carboxylate (2.7 g, 9.40 mmol, 1 eq) and HCl (4 M in dioxane, 15.00 mL, 6.39 eq) in CH2Cl2 (30 mL) was stirred at 20°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain ethyl 2-(4-piperidyloxy)acetate (2.1 g, 9.39 mmol, 99.9% yield, hydrochloric acid) as an off-white solid. Step 2: Preparation of ethyl 2-[[1-(3-benzyloxypropyl)-4-piperidyl]oxy]acetate [ka] To a solution of 3-benzyloxypropyl 4-methylbenzenesulfonate (1.19 g, 3.70 mmol, 1.2 eq) and ethyl 2-(4-piperidyloxy)acetate (690 mg, 3.08 mmol, 1 eq, hydrochloric acid) in DMF (3 mL), K2CO3 (853 mg, 6.17 mmol, 2 eq) was added, and the reaction mixture was stirred at 50°C for 12 hours. Then water (30 mL) was added, and the resulting mixture was extracted with RINKAN (2 x 20 mL). The combined organic extract was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained residue was purified by prep-TLC (CH2Cl2:CH3OH=10:1) with SiO2 to obtain ethyl 2-[[1-(3-benzyloxypropyl)-4-piperidyl]oxy]acetate (570 mg, 1.70 mmol, 55.09% yield) as a colorless oil. 1H-NMR (400 MHz, CDCl3) δ 7.27 (s, 5H), 4.44 (s, 2H), 4.22 - 4.12 (m, 2H), 4.06 - 3.98 (m, 2H), 3.51 - 3.28 (m, 3H), 2.77 - 2.66 (m, 2H), 2.43 - 2.29 (m, 2H), 2.07 (t, J = 9.7 Hz, 2H), 1.93 - 1.82 (m, 2H), 1.80 - 1.68 (m, 2H), 1.61 (dtd, J = 3.6, 9.3, 12.8 Hz, 2H), 1.23 (t, J = 7.1 Hz, 3H). Step 3: Preparation of methyl 2-[[1-(3-hydroxypropyl)-4-piperidyl]oxy]acetate [ka] A solution of ethyl 2-[[1-(3-benzyloxypropyl)-4-piperidyl]oxy]acetate (570 mg, 1.70 mmol, 1 eq) in CH3OH (10 mL) was mixed with Pd / C (50 mg, 5%) under an N2 atmosphere. The resulting suspension was degassed under vacuum and purged several times with H2. The resulting mixture was then stirred at 50°C for 12 hours under H2 (50 psi). The reaction product was filtered, and the filtrate was concentrated under reduced pressure to obtain crude methyl 2-[[1-(3-hydroxypropyl)-4-piperidyl]oxy]acetate (400 mg) as a colorless oil. Step 4: Preparation of tert-butyl 4-[7-bromo-6-chloro-8-fluoro-2-[3-[4-(2-methoxy-2-oxo-ethoxy)-1-piperidyl]propoxy]quinazoline-4-yl]piperazine-1-carboxylate [ka] A mixture of methyl 2-[[1-(3-hydroxypropyl)-4-piperidyl]oxy]acetate (217 mg, 0.94 mmol, 1.5 eq), tert-butyl 4-(7-bromo-2,6-dichloro-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate (300 mg, 0.62 mmol, 1 eq), Cs2CO3 (407 mg, 1.25 mmol, 2 eq), and 1,4-diazabicyclo[2.2.2]octane (21 mg, 0.19 mmol, 0.02 mL, 0.3 eq) in CH3CN (20 mL) was stirred at 50°C for 2 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by preparative reverse-phase HPLC (30-60% CH3CN in water (0.225% formic acid)) to obtain butyl 4-[7-bromo-6-chloro-8-fluoro-2-[3-[4-(2-methoxy-2-oxoethoxy)-1-piperidyl]propoxy]quinazolin-4-yl]piperazine-1-carboxylate (100 mg, 0.15 mmol, 23.7% yield) as a colorless oil. LC / MS (ESI) m / z: 676.2[M+H] + . Step 5: Preparation of tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[3-[4-(2-methoxy-2-oxoethoxy)-1-piperidyl]propoxy]quinazoline-4-yl]piperazine-1-carboxylate [ka] THF (20 mL) contains tert-butyl 4-[7-bromo-6-chloro-8-fluoro-2-[3-[4-(2-methoxy-2-oxo-ethoxy)-1-piperidyl]propoxy]quinazolin-4-yl]piperazine-1-carboxylate (100 mg, 0.15 mmol, 1 eq) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalene-2-ol ( To a solution of 40 mg, 0.15 mmol, 1 eq) of [unclear], K3PO4 (1.5 M, 0.3 mL, 3 eq) and methanesulfonate (2-dicyclohexylphosphino-2,4,6-tri-i-propyl-1,1-biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(II) (12.54 mg, 0.02 mmol, 0.1 eq) were added, and the reaction mixture was stirred at 50°C for 12 hours under an N2 atmosphere. Then, water (30 mL) was added, and the resulting mixture was extracted with ELISA (2 x 20 mL). The combined organic extract was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained residue was purified by prep-TLC (CH2Cl2:CH3OH=10:1) in SiO2 to obtain tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[3-[4-(2-methoxy-2-oxo-ethoxy)-1-piperidyl]propoxy]quinazoline-4-yl]piperazine-1-carboxylate (55 mg, 0.07 mmol, 45.8% yield, 91% purity) as a yellow oil. LC / MS (ESI) m / z: 738.3[M+H] + . Step 6: Preparation of 2-[[1-[3-[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]acetic acid [ka] Solutions of tert-butyl 4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-2-[3-[4-(2-methoxy-2-oxo-ethoxy)-1-piperidyl]propoxy]quinazoline-4-yl]piperazine-1-carboxylate (55 mg, 0.07 mmol, 1 eq) and LiOH (9 mg, 0.22 mmol, 3 eq) in water (0.5 mL), THF (0.5 mL), and CH3OH (0.5 mL) were stirred at 20°C for 0.5 hours. The mixture was concentrated under reduced pressure to obtain crude 2-[[1-[3-[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]acetic acid (53 mg, 0.07 mmol, 98.2% yield) as a colorless oil. Step 8: Preparation of tert-butyl 4-[6-chloro-8-fluoro-2-[3-[4-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]-1-piperidyl]propoxy]-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]piperazine-1-carboxylate [ka] (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (35 mg, 0.07 mmol, 1 eq, hydrochloric acid) and 2-[[1-[3-[4-(4-tert-butoxycarbonylpiperazine-1-yl)-6 To a solution of -chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]acetic acid (53 mg, 0.07 mmol, 1 eq), HATU (56 mg, 0.15 mmol, 2 eqs) and diisopropylethylamine (28 mg, 0.22 mmol, 0.04 mL, 3 eqs) were added, and the reaction mixture was stirred at 25°C for 0.5 hours. Then water (30 mL) was added, and the resulting mixture was extracted with RINKAN (2 x 20 mL). The combined organic extract was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by prep-TLC (CH2Cl2:CH3OH=9:1) in SiO2 to obtain tert-butyl 4-[6-chloro-8-fluoro-2-[3-[4-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]-1-piperidyl]propoxy]-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]piperazine-1-carboxylate (47 mg, 0.04 mmol, 55.8% yield) as a colorless oil. LC / MS (ESI) m / z: 1150.6[M+H] + . Step 9: Preparation of (2S,4R)-1-[(2S)-2-[[2-[[1-[3-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1-ylquinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] To a solution of tert-butyl 4-[6-chloro-8-fluoro-2-[3-[4-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]-1-piperidyl]propoxy]-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]piperazine-1-carboxylate (47 mg, 0.04 mmol, 1 eq) in CH2Cl2 (7 mL), TFA was added, and the reaction mixture was stirred at 20°C for 0.5 hours. The mixture was concentrated under reduced pressure to obtain (2S,4R)-1-[(2S)-2-[[2-[[1-[3-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1-ylquinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (47 mg, 0.04 mmol, 99% yield, TFA salt) as a colorless oil. LC / MS (ESI) m / z: 1050.4[M+H] + . Step 10: Preparation of (2S,4R)-1-[(2S)-2-[[2-[[1-[3-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] (2S,4R)-1-[(2S)-2-[[2-[[1-[3-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1-ylquinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5- To a solution of propyl-2-enoyl chloride (3.29 mg, 0.04 mmol, 0.003 mL, 0.9 eq) in CH2Cl2 (1 mL) was added dropwise to a solution of phenylethyl pyrrolidine-2-carboxamide (47 mg, 0.04 mmol, 1 eq, trifluoroacetic acid) and 2,6-lutidine (130 mg, 1.21 mmol, 0.14 mL, 30 eq), and the reaction mixture was stirred at -78°C for 30 minutes. Water (30 mL) was then added, and the resulting mixture was extracted with CH2Cl2 (2 x 20 mL). The combined organic extract was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained residue was purified by semi-preparative reverse-phase HPLC (20-50% CH3CN in water (0.225% formic acid)) to obtain (2S,4R)-1-[(2S)-2-[[2-[[1-[3-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazoline-2-yl]oxypropyl]-4-piperidyl]oxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (7.47 mg, 0.006 mmol, 15% yield, 94% purity, formic acid) as a white solid. LC / MS(ESI)m / z:1150.3[M+H] + . 1H-NMR (400 MHz, DMSO-d6) δ (s, 1H), 8.45 (d, J = 7.7 Hz, 1H), 8.37 (s, 1H), 8.01 (s, 1H), 7.80 (d, J = 8.3 Hz, 1H), 7.49 - 7.40 (m, 3H), 7.39 - 7.26 (m, 4H), 7.25 - 7.18 (m, 2H), 7.07 (d, J = 2.2 Hz, 1H), 6.83 (dd, J = 10.3, 16.7 Hz, 1H), 6.18 (dd, J = 2.4, 16.8 Hz, 1H), 5.79 - 5.69 (m, 1H), 4.88 (t, J = 7.2 Hz, 1H), 4.52 (d, J = 9.7 Hz, 1H), 4.48 - 4.32 (m, 3H), 4.27 (s, 1H), 4.02 - 3.46 (m, 12H), 2.75 - 2.68 (m, 1H), 2.53 - 2.52 (m, 2H), 2.45 (s, 3H), 2.44 - 2.39 (m, 3H), 2.33 (s, 1H), 2.13 - 2.02 (m, 3H), 1.93 - 1.74 (m, 5H), 1.52 - 1.43 (m, 2H), 1.36 (d, J = 7.1 Hz, 3H), 0.92 (s, 9H)。

[0245] (2S,4R)-1-[(2S)-2-[[2-[[1-[(1R,2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazoline-2-yl]oxycyclopentyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide and (2 Exemplary synthesis of S,4R)-1-[(2S)-2-[[2-[[1-[(1S,2S)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazoline-2-yl]oxycyclopentyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide Step 1: Preparation of benzyl 4-((2-(tert-butoxy)-2-oxoethoxy)methyl)piperidine-1-carboxylate [ka] To a mixture of benzyl 4-(hydroxymethyl)piperidine-1-carboxylate (10 g, 40.1 mmol, 1 eq) in THF (50 mL), NaH (3.2 g, 80.2 mmol, 60%, 2 eq) was gradually added, followed by tert-butyl 2-bromoacetate (15.7 g, 80.2 mmol, 2 eq). The reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was quenched with additional water (10 mL), and the resulting mixture was extracted with ELISA (3 x 50 mL). The combined organic extract was washed with brine (3 x 50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The resulting residue was purified by silica gel chromatography (10-20% ethyl ether in petroleum ether) to obtain benzyl 4-[(2-tert-butoxy-2-oxo-ethoxy)methyl]piperidine-1-carboxylate (3.6 g, 9.91 mmol, 25% yield) as a colorless oil.1 H-NMR (400 MHz, CDCl3) δ 7.42 - 7.30 (m, 4H), 5.14 (s, 2H), 4.19 (d, J = 18.2 Hz, 2H), 3.96 (s, 2H), 3.38 (d, J = 6.4 Hz, 2H), 2.81 (s, 2H), 1.79 (d, J = 13.2 Hz, 3H), 1.53 - 1.47 (m, 9H), 1.26 - 1.13 (m, 2H). Step 2: Preparation of tert-butyl 2-(piperidine-4-ylmethoxy)acetate [ka] A solution of benzyl 4-[(2-tert-butoxy-2-oxo-ethoxy)methyl]piperidine-1-carboxylate (3.1 g, 8.53 mmol, 1 eq) and NH4OH (107 mg, 0.85 mmol, 28% purity, 0.1 eq) in CH3OH (30 mL) was mixed with Pd / C (300 mg, 10%) under an N2 atmosphere. The resulting suspension was degassed under vacuum and purged several times with H2. The reaction mixture was then stirred at 25°C for 16 hours under H2 (15 psi). The mixture was filtered, and the filtrate was concentrated under vacuum. The resulting residue was purified by silica gel chromatography (5-10% CH3OH in CH2Cl2) to obtain tert-butyl 2-(4-piperidylmethoxy)acetate (1.8 g, 7.85 mmol, 92% yield) as a colorless oil. 1 H-NMR (400 MHz, DMSO-d6) δ 3.93 (s, 2H), 3.48 - 3.34 (m, 3H), 3.26 (d, J = 6.0 Hz, 2H), 2.94 (d, J = 12.0 Hz, 2H), 1.60 (d, J = 12.0 Hz, 3H), 1.45 - 1.40 (m, 9H), 1.18 - 0.95 (m, 2H). Step 3: Preparation of tert-butyl 2-((1-((1S,2S)-2-hydroxycyclopentyl)piperidine-4-yl)methoxy) acetate and tert-butyl 2-((1-((1R,2R)-2-hydroxycyclopentyl)piperidine-4-yl)methoxy) acetate [ka] 6-oxabicyclo[3.1.0]hexane (1.3g, 15.7 mmol, 2eq) was added to tert-butyl 2-(4-piperidyl methoxy) acetate (1.8g, 7.85 mmol, 1eq) in ethanol (20 mL), and the reaction mixture was stirred at 80°C for 3 hours. The mixture was concentrated under vacuum, and the resulting residue was purified by silica gel chromatography (10% CH3OH in CH2Cl2) to obtain tert-butyl 2-[[1-(2-hydroxycyclopentyl)-4-piperidyl]methoxy] acetate (1.8g, 5.74 mmol, 73% yield) as a yellow oil. 1 H-NMR (400 MHz, DMSO-d6) δ 4.50 (s, 1H), 3.98 - 3.83 (m, 3H), 3.33 - 3.24 (m, 3H), 3.14 - 3.04 (m, 1H), 2.92 - 2.80 (m, 1H), 2.43 - 2.34 (m, 1H), 2.07 - 1.85 (m, 2H), 1.83 - 1.70 (m, 2H), 1.67 - 1.51 (m, 4H), 1.43 (s, 9H), 1.41 - 1.38 (m, 2H), 1.22 - 1.08 (m, 2H). Step 4: Preparation of tert-butyl 4-(7-bromo-2-(((1R,2R)-2-(4-((2-(tert-butoxy)-2-oxoethoxy)methyl)piperidine-1-yl)cyclopentyl)oxy)-6-chloro-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate and tert-butyl 4-(7-bromo-2-(((1S,2S)-2-(4-((2-(tert-butoxy)-2-oxoethoxy)methyl)piperidine-1-yl)cyclopentyl)oxy)-6-chloro-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate [ka] To a mixture of tert-butyl 4-(7-bromo-2,6-dichloro-8-fluoroquinazolin-4-yl)piperazine-1-carboxylate (3.5 g, 7.18 mmol, 1.5 eq), tert-butyl 2-[[1-(2-hydroxycyclopentyl)-4-piperidyl]methoxy]acetate (1.5 g, 4.79 mmol, 1 eq), and 1,4-diazabicyclo[2.2.2]octane (54 mg, 0.48 mmol, 0.1 eq) in CH3CN (10 mL), Cs2CO3 (3.1 g, 9.57 mmol, 2 eq) was added, and the reaction mixture was stirred at 50°C for 16 hours. The mixture was filtered, and the filtrate was concentrated under vacuum. The obtained residue was purified by prep-TLC (50% ethyl acetate in petroleum ether) followed by prep-HPLC (75-100% CH3CN in water (10 mM NH4HCO3)). Further purification by SFC yielded tert-butyl 4-[7-bromo-2-[(1R,2R)-2-[4-[(2-tert-butoxy-2-oxo-ethoxy)methyl]-1-piperidyl]cyclopentoxy]-6-chloro-8-fluoroquinazolin-4-yl]piperazine-1-carboxylate (400 mg, 0.53 mmol, 11% yield) as a yellow solid and tert-butyl 4-[7-bromo-2-[(1S,2S)-2-[4-[(2-tert-butoxy-2-oxo-ethoxy)methyl]-1-piperidyl]cyclopentoxy]-6-chloro-8-fluoroquinazolin-4-yl]piperazine-1-carboxylate (400 mg, 0.53 mmol, 11% yield) as a yellow solid. tert-butyl4-[7-bromo-2-[(1R,2R)-2-[4-[(2-tert-butoxy-2-oxo-ethoxy)methyl]-1-piperidyl]cyclopentoxy]-6-chloro-8-fluoroquinazolin-4-yl]piperazine-1-carboxylate:SFC RT=2.021 min tert-butyl4-[7-bromo-2-[(1S,2S)-2-[4-[(2-tert-butoxy-2-oxo-ethoxy)methyl]-1-piperidyl]cyclopentoxy]-6-chloro-8-fluoroquinazolin-4-yl]piperazine-1-carboxylate:SFC RT=2.176 min Step 5: Preparation of tert-butyl 4-[2-[(1R,2R)-2-[4-[(2-tert-butoxy-2-oxo-ethoxy)methyl]-1-piperidyl]cyclopentoxy]-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]piperazine-1-carboxylate [ka] THF (10 mL) contains tert-butyl 4-[7-bromo-2-[(1R,2R)-2-[4-[(2-tert-butoxy-2-oxo-ethoxy)methyl]-1-piperidyl]cyclopentoxy]-6-chloro-8-fluoroquinazolin-4-yl]piperazine-1-carboxylate (400 mg, 0.53 mmol, 1 eq), methanesulfonate (2-dicyclohexylphosphino-2,4,6-tri-i-p A mixture of palladium(II)(2-amino-1,1-biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(II) (22 mg, 0.03 mmol, 0.05 eq) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-ol (128 mg, 0.48 mmol, 0.9 eq) was mixed with K3PO4 (1.5 N in water, 3 eq) under nitrogen, and the reaction mixture was stirred at 50°C for 16 hours. Water (30 mL) was then added, and the resulting mixture was extracted with ELISA (3 x 20 mL). The combined organic extract was washed with brine (3 x 20 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The obtained residue was purified by prep-TLC (10% CH3OH in CH2Cl2) to obtain tert-butyl 4-[2-[(1R,2R)-2-[4-[(2-tert-butoxy-2-oxo-ethoxy)methyl]-1-piperidyl]cyclopentoxy]6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]piperazine-1-carboxylate (210 mg, 0.26 mmol, 48% yield) as a yellow solid. LC / MS (ESI) m / z: 820.4[M+H] + . Step 6: Preparation of 2-[[1-[(1R,2R)-2-[4-(4-tert-butoxycarbonylpiperazine-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-2-yl]oxycyclopentyl]-4-piperidyl]methoxy]acetic acid [ka] A mixture of tert-butyl 4-[2-[(1R,2R)-2-[4-[(2-tert-butoxy-2-oxo-ethoxy)methyl]-1-piperidyl]cyclopentoxy]-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]piperazine-1-carboxylate (150 mg, 0.18 mmol, 1 eq) and LiOH (252 mg, 6 mmol, 33 eq) in CH3OH (2 mL), H2O (2 mL), and THF (2 mL) was stirred at 25°C for 1 hour. The pH of the reaction mixture was adjusted to 6 by adding 1H HCl (10 mL), and the resulting mixture was extracted with ELISA (3 x 30 mL). The combined organic extracts were washed with brine (3 x 30 mL), dried over anhydrous sodium 2SO4, filtered, and concentrated to obtain 2-[[1-[(1R,2R)-2-[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-2-yl]oxycyclopentyl]-4-piperidyl]methoxy]acetic acid (130 mg, 0.17 mmol, 93% yield) as a yellow solid. LC / MS MS(ESI) m / z: 764.4[M+H] + . Step 7: Preparation of tert-butyl 4-[6-chloro-8-fluoro-2-[(1R,2R)-2-[4-[[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]methyl]-1-piperidyl]cyclopentoxy]-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]piperazine-1-carboxylate [ka] 2-[[1-[(1R,2R)-2-[4-(4-tert-butoxycarbonylpiperazin-1-yl)-6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazoline-2-yl]oxycyclopentyl]-4-piperidyl]methoxy]acetic acid (130 mg, 0.17 mmol, 1 eq) in DMF (10 mL), (2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-meth A mixture of luchiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (98 mg, 0.20 mmol, 1.2 eq, hydrochloride), 1-hydroxybenzotriazole (46 mg, 0.34 mmol, 2 eq), and N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (65 mg, 0.34 mmol, 2 eq) was mixed with diisopropylethylamine (66 mg, 0.51 mmol, 3 eq), and the reaction mixture was stirred at 25°C for 16 hours. Then water (20 mL) was added, and the resulting mixture was extracted with RINKAN (3 x 30 mL). The combined organic extract was washed with brine (3 x 30 mL), dried over anhydrous sodium 2SO4, filtered, and concentrated. The resulting residue was purified by prep-TLC (10% CH3OH in CH2Cl2) to obtain tert-butyl 4-[6-chloro-8-fluoro-2-[(1R,2R)-2-[4-[[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]methyl]-1-piperidyl]cyclopentoxy]-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]piperazine-1-carboxylate (30 mg, 0.03 mmol, 15% yield) as a yellow solid. LC / MS (ESI) m / z: 1190.6[M+H] + . Step 8: Preparation of (2S,4R)-1-[(2S)-2-[[2-[[1-[(1R,2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1-ylquinazoline-2-yl]oxycyclopentyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] Trifluoroacetic acid (1.5 g, 13.5 mmol, 536 eq) was added to tert-butyl 4-[6-chloro-8-fluoro-2-[(1R,2R)-2-[4-[[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo-ethoxy]methyl]-1-piperidyl]cyclopentoxy]-7-(3-hydroxy-1-naphthyl)quinazoline-4-yl]piperazine-1-carboxylate (30 mg, 0.03 mmol, 1 eq) in CH2Cl2 (5 mL), and the reaction mixture was stirred at 25 °C for 0.5 hours. The mixture was concentrated to obtain (2S,4R)-1-[(2S)-2-[[2-[[1-[(1R,2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1-ylquinazoline-2-yl]oxycyclopentyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (30 mg, 0.02 mmol, 99% yield, trifluoroacetate) as a yellow solid. LC / MS (ESI) m / z: 1090.5[M+H] + . Step 9: Preparation of (2S,4R)-1-[(2S)-2-[[2-[[1-[(1R,2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazoline-2-yl]oxycyclopentyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide [ka] (2S,4R)-1-[(2S)-2-[[2-[[1-[(1R,2R)-2-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)-4-piperazine-1-ylquinazoline-2-yl]oxycyclopentyl]-4-piperidyl]methoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[( A mixture of 1S)-1-[4-(4-methylthiazole-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (30 mg, 0.02 mmol, 1 eq, trifluoroacetate) and 2,6-lutidine (27 m...

Claims

[Claim 1] The invention described in the specification.

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