Chemicals that target chromatin remodeling complexes and their use

Abstract

This disclosure relates to compounds, compositions, and methods for treating diseases or conditions mediated by abnormal bromodomain PHD finger transcription factor (BPTF) activity.

Description

[Technical Field]

[0001] (Cross-reference of related applications) This application claims the benefit of U.S. Provisional Patent Application No. 63 / 541,946, filed on 2 October 2023 under Section 119(e) of the U.S. Patent Act, which is incorporated herein by reference in its entirety.

[0002] (support) The project leading to this application is funded by the Innovative Medicines Initiative 2 Joint Undertaking (JU) under Grant Agreement No. 875510. The JU is supported by the European Union's Horizon 2020 Research and Innovation Programme, as well as EFPIA, and KUNGLIGA TEKNISKA HOEGSKOLAN, DIAMOND LIGHT SOURCE LIMITED, Ontario Institute for Cancer Research, and ROYAL INSTITUTION FOR THE ADVANCEMENT OF LEARNING MCGILL UNIVERSITY. [Background technology]

[0003] Bromodomain PHD finger transcription factor (BPTF), a member of the chromatin remodeler imitation switch (ISWI) family, is mutated or overexpressed in multiple oncological types. Chromatin modifications play a major role in tumorigenesis, chemotherapy resistance, and susceptibility to immune responses.

[0004] To date, no chemical probes that selectively degrade BPTF have been reported. Reported BPTF small molecule inhibitors have either exhibited off-target cytotoxic effects or have been shown to be ineffective in preventing tumor cell proliferation. Therefore, there is a need for compounds that provide effective inhibition of tumor cell proliferation while minimizing off-target cytotoxic effects in cancer treatment. [Overview of the Initiative] [Means for solving the problem]

[0005] A first aspect of this disclosure is a compound having a structure represented by formula I or II:

[0006] [ka] During the ceremony Degron binds to cerebron (CRBN) or von Hippel-Lindau (VHL) tumor suppressor. The focus is on compounds in which the linker represents the part that covalently bonds degron and a nitrogen atom, or their pharmaceutically acceptable salts or stereoisomers.

[0007] Another aspect of this disclosure relates to a pharmaceutical composition comprising a compound of formula I or II, or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier.

[0008] Another aspect of this disclosure provides a method for preparing compounds of formula (I) or (II).

[0009] Further aspects of this disclosure relate to methods for treating cancers with abnormal BPTF activity, comprising administering a compound of formula I or II, or a pharmaceutically acceptable salt or stereoisomer thereof, to a subject as needed. [Brief explanation of the drawing]

[0010] [Figure 1A] This is a heat map. [Figure 1B] This scatter plot shows that dBPTF-2 is selective for BPTF chemoproteomics using the cell line K562. [Figure 2A]This is an immunoblot showing the degradation of BPTF by the compound at the indicated concentrations. Figure 2A shows NVS-BPTF-1, dBPTF-1, dBPTF-2, and dBPTF-3 using the VAL cell line after 24 hours. [Figure 2B] This is an immunoblot showing the degradation of BPTF by the compound at the indicated concentrations. Figure 2B shows dBPTF-2 using the VAL cell line after 8 hours. [Figure 2C] This is an immunoblot showing the degradation of BPTF by the compound at the indicated concentrations. Figure 2C shows dBPTF-2, dBPTF-4, dBPTF-5, and dBPTF-6 using the VAL cell line after 24 hours. [Figure 2D] This is an immunoblot showing the degradation of BPTF by the compound at the indicated concentrations. Figure 2D shows dBPTF-2, dBPTF-6, dBPTF-7, and dBPTF-8 using the VAL cell line after 24 hours. [Figure 2E] This is an immunoblot showing the degradation of BPTF by the compound at the indicated concentrations. Figure 2E shows dBPTF-2 and dBPTF-7 using the VAL cell line after 6 hours. [Figure 2F] This is an immunoblot showing the degradation of BPTF by the compound at the indicated concentrations. Figure 2F shows dBPTF-2, dBPTF-9, dBPTF-10, and dBPTF-11 using the VAL cell line after 24 hours. [Figure 2G] This is an immunoblot showing the degradation of BPTF by the compound at the indicated concentrations. Figure 2G shows NVS-BPTF-1, dBPTF-2, dBPTF-12, and dBPTF-13 using the VAL cell line after 24 hours. [Figure 3A] This immunoblot demonstrates that CRBN knockout rescues degradation. Figure 3A shows BPTF degradation of dBPTF-2, dBPTF-9, and dBPTF-11 using cell line 293T after 24 hours. [Figure 3B]This immunoblot demonstrates that CRBN knockout rescues degradation. Figure 3B shows that dBPTF-2, dBPTF-9, and dBPTF-11 do not degrade BPTF after 24 hours when using cell line 293T CRBN KO. [Figure 4A] This is an immunoblot showing BPTF degradation in different cancer types. Figure 4A shows BPTF degradation of dBPTF-2, dBPTF-9, and dBPTF-11 using the MCF7 (milk) cell line after 24 hours. [Figure 4B] This is an immunoblot showing BPTF degradation in different cancer types. Figure 4B shows BPTF degradation of dBPTF-2, dBPTF-9, and dBPTF-11 using the NCIH2030 (lung) cell line after 24 hours. [Figure 5A] (6 hours) This graph shows the formation of ternary complexes with various compounds using 293T cells transfected with NanoLuc®-tagged BPTF and HaloTag®-tagged CRBN. [Figure 5B] This graph shows the formation of ternary complexes with various compounds using 293T cells transfected with NanoLuc®-tagged BPTF and HaloTag®-tagged CRBN (20 hours). [Figure 6A] This shows that NVS-BPTF-1B degraded BPTF in HiBiT cells using cell line 293T. Figure 6A is a graph showing the detection of Nano-Glo® HiBit lysis after 6 hours. [Figure 6B] This shows that NVS-BPTF-1B degraded BPTF in HiBiT cells using cell line 293T. Figure 6B is a graph showing the detection of Nano-Glo® HiBit lysis after 20 hours. [Figure 6C] This shows that NVS-BPTF-1B degraded BPTF in HiBiT cells using cell line 293T. Figure 6C is a graph showing cell viability after 20 hours. [Figure 7A]This demonstrates that NVS-BPTF-1B functioned consistently throughout the assay. Figure 7A shows immunoblots of BPTF degradation after 8 hours using the cell line VAL, with NVS-BPTF-1B, TP-238-2, and TP-238-3. [Figure 7B] This demonstrates that NVS-BPTF-1B functioned consistently throughout the assay. Figure 7B is a graph showing Nano-Glo® HiBit lysis detection after 6 hours. [Figure 8A] This shows that NVS-BPTF-1B degraded BPTF at an early stage. Figure 8A is an immunoblot showing BPTF degradation at 2, 4, 6, and 8 hours using the VAL cell line. [Figure 8B] This indicates that NVS-BPTF-1B degraded BPTF at an early stage. Figure 8B is a bar graph showing the %BPTF band density at the indicated time and concentration. [Figure 9A] This shows the rescue of BPTF degradation via NVS-BPTF-1B using cell line 293T. Figure 9A is an immunoblot showing BPTF degradation after 8 hours in a series of rescue experiments. [Figure 9B] This demonstrates BPTF degradation rescue via NVS-BPTF-1B using cell line 293T. Figure 9B is a bar graph showing the %BPTF band density. [Figure 10] NVS-BPTF-1B is an immunoblot that demonstrates the lift-off of BPTF from histones using the VAL cell line at the indicated time. [Figure 11A] This shows that BPTF remains present in the nucleus for more than 24 hours. Figure 11A is an immunoblot showing the presence of BPTF after 8 hours, using the VAL cell line. [Figure 11B] This shows that BPTF remains present in the nucleus for more than 24 hours. Figure 11B is an immunoblot showing the presence of BPTF after 24 hours, using the VAL cell line. [Figure 12]This immunoblot shows that BPTF protein levels begin to recover during compound washout. [Figure 13] This is a series of graphs that identify cell types that are sensitive to BPTF loss. [Figure 14A] This study demonstrates that the PAX3-FOXO1-dependent cell line RH4 is more susceptible to BPTF loss. [Figure 14B] This study demonstrates that the PAX3-FOXO1-dependent cell line RH4 is more susceptible to BPTF loss. [Figure 15A] This demonstrates that NURF and PAX3-FOXO1 physically interact. [Figure 15B] This demonstrates that NURF and PAX3-FOXO1 physically interact. [Figure 16A] This shows BPTF degradation in PAX3-FOXO1 fusion-dependent cells. Figure 16A is an immunoblot showing NVS-BPTF-1 and NVS-BPTF-1-B after 48 hours using the RH4 cell line. [Figure 16B] This shows BPTF degradation in PAX3-FOXO1 fusion-dependent cells. Figure 16B is an immunoblot showing NVS-BPTF-1 and NVS-BPTF-1-B after 48 hours, using the RH4 cell line and histone extracts. [Figure 17] This is a series of graphs showing that PAX3 / 7-FOXO1-dependent cells are sensitive to BPTF protein loss. [Figure 18A] This demonstrates that NVS-BPTF-1B can lift off the fusion protein from histones at a later time point. Figure 18A shows immunoblots of NVS-BPTF-1 and NVS-BPTF-1-B using cell line RH30 after 48 hours. [Figure 18B] This demonstrates that NVS-BPTF-1B can lift off the fusion protein from histones at a later time point. Figure 18B shows immunoblots of NVS-BPTF-1 and NVS-BPTF-1-B using cell line RH30 after 72 hours. [Figure 19A] This study demonstrates that functional loss of BPTF due to shRNA knockdown is accompanied by protein loss of PAX3-FOXO1. [Figure 19B] This study demonstrates that functional loss of BPTF due to shRNA knockdown is accompanied by protein loss of PAX3-FOXO1. [Figure 20] This is an immunoblot showing BPTF degradation by dBPTF-2, dBPTF-9, and dBPFT-11 using the RH4 cell line 24 hours later. [Figure 21] This is an immunoblot showing BPTF degradation and a decrease in PAX3-FOXO1 protein levels after 24 hours using the RH30 cell line, mediated by dBPTF-2, dBPTF-9, and dBPFT-11. [Figure 22A] This immunoblot demonstrates that CRBN knockout rescues degradation. Figure 22A shows dBPTF-2-mediated BPTF degradation using the RH4 cell line after 72 hours. [Figure 22B] This immunoblot demonstrates that CRBN knockout rescues degradation. Figure 22B shows that dBPTF-2 does not degrade BPTF after 72 hours when using the RH4 CRBN KO cell line. [Figure 23] This is a scatter plot of proteomic studies showing the selective degradation of dBPTF-9. [Figure 24] This graph shows that dBPTF-2, dBPTF-9, and dBPTF-11 are more potent inhibitors than NVS-BPTF-1 after 6 days using the RH4 cell line. [Figure 25] This shows a colony formation assay using RMS cell lines (fusion-negative, fusion-positive, and fusion-positive but lacking CRBN) treated with NVS-BPTF-1 and NVS-BPTF-1-B in gradually increasing doses for 14 days. [Figure 26A] This shows the cell viability of RH4 and RH28 fusion-positive RMS cell lines after treatment with BPTF-targeted compounds. [Figure 26B]This shows the cell viability of RH4 and RH28 fusion-positive RMS cell lines after treatment with BPTF-targeted compounds. [Figure 27A] This immunoblot shows the effect of dBPTF-9 on fusion protein levels on histones. Figure 27A shows dBPTF-9-mediated BPTF degradation using the RH4 cell line after 8 hours. [Figure 27B] This is an immunoblot showing the effect of dBPTF-9 on fusion protein levels on histones. Figure 27B shows dBPTF-9-mediated BPTF degradation in the RH4 cell line after 24 hours. [Figure 28A] This section presents xenograft studies of tumor-bearing mice treated with either a vehicle control or dBPTF-9. Figure 28A is a graph showing that treatment with dBPTF-9 slowed tumor progression. [Figure 28B] This section presents xenotransplant studies of tumor-bearing mice treated with either vehicle control or dBPTF-9. Figure 28B is a graph showing that treatment with dBPTF-9 extended animal survival. [Figure 28C] This section presents xenograft studies of tumor-bearing mice treated with either a vehicle control or dBPTF-9. Figure 28C is a graph showing that treatment with dBPTF-9 was well tolerated in mice and that the mice did not experience any change in body weight compared to the control group. [Figure 29A] This shows the selective degradation of dBPTF-13. Figure 29A is a graph showing the dissolution detection of the indicated compound using Nano-Glo® HiBit. [Figure 29B] This shows the selective degradation of dBPTF-13. Figure 29B is a graph showing the Nano-Glo® HiBit dissolution detection for the compound shown. [Figure 29C] This shows the selective degradation of dBPTF-13. Figure 29C is a graph showing the Nano-Glo® HiBit dissolution detection of dBPTF-13 at the indicated time. [Figure 29D]This shows the selective degradation of dBPTF-13. Figure 29D is a graph showing the dissolution detection of the indicated compound using Nano-Glo® HiBit. [Figure 29E] This shows the selective decomposition of dBPTF-13. Figure 29E is a scatter plot of proteome studies showing the selective decomposition of dBPTF-13. [Figure 29F] This shows the selective decomposition of dBPTF-13. Figure 29F is a model illustrating the binding interaction between dBPTF-13 and BPTF. [Figure 29G] This shows the selective decomposition of dBPTF-13. Figure 29G is a model illustrating the binding interaction between dBPTF-13 and BPTF. [Figure 30A] This shows the selective degradation of dBPTF-2. Figure 30A is a graph showing the dissolution detection of the indicated compound using Nano-Glo® HiBit. [Figure 30B] This shows the selective degradation of dBPTF-2. Figure 30B is a graph showing the dissolution detection of the indicated compound using Nano-Glo® HiBit. [Figure 30C] This shows the selective degradation of dBPTF-2. Figure 30C is a graph showing the Nano-Glo® HiBit dissolution detection of dBPTF-2 at the indicated time. [Figure 30D] This shows the selective degradation of dBPTF-2. Figure 30D is a graph showing the dissolution detection of the indicated compound using Nano-Glo® HiBit. [Figure 30E] This shows the selective degradation of dBPTF-2. Figure 30E is a scatter plot of a proteome study showing the selective degradation of dBPTF-13 at 8 hours and 24 hours. [Figure 30F] This shows the selective decomposition of dBPTF-2. Figure 30F is a series of bar graphs showing that dBPTF-2 decomposes BPTF, STRAP, and CSDE1. [Figure 30G]This shows the selective degradation of dBPTF-2. Figure 30G is a series of Western blots showing that dBPTF-2 degrades STRAP and CSDE1. [Figure 31] This graph shows the Nano-Glo(registered trademark) HiBit dissolution detection results and corresponding tabular data for the indicated compounds. [Figure 32] This graph shows the Nano-Glo(registered trademark) HiBit dissolution detection results and corresponding tabular data for the indicated compounds. [Modes for carrying out the invention]

[0011] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art of the field to which the subject matter of this specification pertains. Where used herein and in the appended claims, unless otherwise specified, the following terms have the meanings provided for the convenience of understanding this disclosure.

[0012] As used herein and in the appended claims, the singular forms “a,” “an,” and “the” refer to multiple subjects unless the context clearly indicates otherwise. For example, a reference to “a composition” includes mixtures of two or more such compositions, and a reference to “an inhibitor” includes mixtures of two or more such inhibitors, and so on.

[0013] Unless otherwise specified, the term "approximately" means within 10% of the specific value modified by the term "approximately," for example, within 5%, 2%, or 1%.

[0014] The transitional phrase "comprising," which is synonymous with "including," "containing," or "characterized by," is comprehensive or non-restrictive and does not exclude additional unlisted elements or method steps. When used in the context of the number of heteroatoms in a heterocyclic structure, it means that the heterocyclic group has the minimum number of heteroatoms. In contrast, the transitional phrase "consists of" excludes any elements, steps, or components not expressed in the claims. The transitional phrase "essentially consists of" limits the claims to the specific materials or steps of the disclosure "and not substantially affect the basic and novel features(s)."

[0015] With respect to the compounds of this disclosure, and to the extent that the following terms are used herein to further describe them, the following definitions apply.

[0016] As used herein, the term "alkyl" refers to a saturated linear or branched monovalent hydrocarbon group. In some embodiments, the alkyl group is a C1-C6 group. In some embodiments, for any one or more groups of the compound of formula I or II, to the extent not otherwise disclosed, the alkyl group is a C0-C6, C0-C5, C-0-C3, C1-C6, C1-C5, C1-C4, or C1-C3 group (wherein C0 alkyl refers to a bond). Examples of alkyl groups include methyl, ethyl, 1-propyl, 2-propyl, i-propyl, 1-butyl, 2-methyl-1-propyl, 2-butyl, 2-methyl-2-propyl, 1-pentyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, and 3,3-dimethyl-2-butyl. In some embodiments, the alkyl group is a C1-C3 alkyl group. In some embodiments, the alkyl group is a C1-C2 alkyl group. In some embodiments, the alkyl group is a methyl group.

[0017] As used herein, the term "alkylene" refers to a straight or branched divalent hydrocarbon chain consisting only of carbon and hydrogen, containing no unsaturation, and having 1 to 15 carbon atoms, with the rest of the molecule linked to a radical group, such as methylene, ethylene, propylene, n-butylene, etc. The alkylene chain may be bonded to the rest of the molecule via single bonds and to the radical group via single bonds. In some embodiments, the alkylene group contains 1 to 15 carbon atoms (C1-C15). 15 Alkylene). In some embodiments, the alkylene group contains 1 to 12 carbon atoms (C1 to C2). 12 Alkylene). In some embodiments, the alkylene group contains 1 to 10 carbon atoms (C1 to C1). 10Alkylene). In some embodiments, the alkylene group contains 1 to 8 carbon atoms (C1-C8 alkylene). In other embodiments, the alkylene group contains 1 to 5 carbon atoms (C1-C5 alkylene). In other embodiments, the alkylene group contains 1 to 4 carbon atoms (C1-C4 alkylene). In other embodiments, the alkylene group contains 1 to 3 carbon atoms (C1-C3 alkylene). In other embodiments, the alkylene group contains 1 to 2 carbon atoms (C1-C2 alkylene). In other embodiments, the alkylene group contains 1 carbon atom (C1 alkylene).

[0018] As used herein, the term "carbocyclic" (also known as "carbocyclyl") refers to a group used alone or as part of a larger part, containing saturated, partially unsaturated, or aromatic ring systems having 3 to 12 carbon atoms (e.g., alkylcarbocyclic groups). The term carbocyclyl includes monocyclic, dicyclic, tricyclic, fusion, bridging, and spirocyclic systems, as well as combinations thereof. In one embodiment, the carbocyclyl contains 3 to 10 carbon atoms (C3-C 10 In one embodiment, the carbocyclyl contains 3 to 6 carbon atoms (C3 to C6). In one embodiment, the carbocyclyl contains 5 to 6 carbon atoms (C5 to C6). In some embodiments, the carbocyclyl has two rings, C6 to C6. 10 It includes. In another embodiment, carbocyclyl is a spirosystem, C5-C 11 This includes. Representative examples of monocyclic carbocyclyls include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopenta-1-enyl, 1-cyclopenta-2-enyl, 1-cyclopenta-3-enyl, cyclohexyl, 1-cyclohexa-1-enyl, 1-cyclohexa-2-enyl, 1-cyclohexa-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, phenyl, and naphthalene.

[0019] As used herein, the term “heterocyclyl” is used alone or as part of a larger term to refer to a “carbocyclyl” containing saturated, partially unsaturated, or aromatic ring systems in which one or more carbon atoms, e.g., 1, 2, 3, 4, or 5, are replaced by heteroatoms or heteroatom-containing groups (e.g., O, N, N(O), S, S(O), or S(O)2). The term heterocyclyl includes monocyclic, dicyclic, tricyclic, fusion, bridging, and spirocyclic systems, as well as combinations thereof. In some embodiments, heterocyclyl refers to 3- to 12-membered heterocyclyl ring systems. In some embodiments, heterocyclyl refers to saturated ring systems, e.g., 3- to 12-membered saturated heterocyclyl ring systems. In some embodiments, heterocyclyl refers to heteroaryl ring systems, e.g., 5- to 12-membered heteroaryl ring systems. The term heterocyclyl also includes C2-C8 heterocycloalkyls, which are saturated or partially unsaturated monocyclic, bicyclic, or spirocyclic systems containing 2-8 carbon atoms and one or more (e.g., 1, 2, or 3) heteroatoms.

[0020] In some embodiments, the heterocyclyl group comprises 3 to 12 ring atoms and includes monocyclic, dicyclic, tricyclic, and spirocyclic systems, where the ring atoms are carbon and 1 to 5 ring atoms are heteroatoms such as nitrogen, sulfur, or oxygen. In some embodiments, the heterocyclyl comprises a 3 to 7-membered monocyclic ring having one or more heteroatoms selected from O, N, and S. In some embodiments, the heterocyclyl comprises a 4 to 6-membered monocyclic ring having one or more heteroatoms selected from O, N, and S. Representative examples of heterocyclils include oxyranil, azilidinil, thiranil, azetidinil, oxetanil, thietanil, pyrrolidinil, dihydro-1H-pyrrolyl, dihydrofuranil, tetrahydropyranil, dihydrothienyl, tetrahydrothienyl, imidazolidinil, piperidinil, piperazinil, morpholinil, thiomorpholinil, 1,1-dioxo-thiomorpholinil, dihydropyranil, tetrahydropyranil, hexahydrothiopyranil, hexahydropyrimidinil, oxazinil, thiadinil, thioxanil, homopiperazinil, homopiperidinil, azepanil, oxepanil, thiepanil, oxazepinil, oxazepanil, diazepanil, diazepinil Examples include thiazepinyl, thiazepanyl, tetrahydrothiopyranil, oxazolidinyl, thiazolidinyl, isothiazolidinyl, oxazolidinyl, imidazolidinyl, thiadinyl, thiophenyl, oxazinyl, thiadiadinyl, oxadiadinyl, dithiadinyl, dioxazinyl, oxathiadinyl, thiatriazinyl, oxatriazinyl, dithiadiadinyl, imidazolinyl, dihydropyrimidyl, tetrahydropyrimidyl, pyrrolinyl, indolinyl, thiapyranil, 2H-pyranil, 4H-pyranil, dioxanil, pyrazolinyl, pyrazolidinyl, dithianyl, dithiolanil, pyrimidinol, pyrimididionyl, piperadinyl, and piperazinedinoyl.Examples of five-membered heterocyclines containing a sulfur or oxygen atom and 1 to 3 nitrogen atoms include thiazolyl (e.g., thiazole-2-yl), thiadiazolyl (e.g., 1,3,4-thiadiazole-5-yl, and 1,2,4-thiadiazole-5-yl), oxazolyl (e.g., oxazol-2-yl), and oxadiazolyl (e.g., 1,3,4-oxadiazole-5-yl, and 1,2,4-oxadiazole-5-yl). Examples of five-membered heterocyclines containing 2 to 4 nitrogen atoms include imidazolyl (e.g., imidazole-2-yl), triazolyl (e.g., 1,3,4-triazol-5-yl, 1,2,3-triazol-5-yl, and 1,2,4-triazol-5-yl), and tetrazolyl (e.g., 1H-tetrazole-5-yl). Examples of six-membered heterocyclyls containing 1 to 3 nitrogen atoms and optionally sulfur or oxygen atoms include pyridyl (e.g., pyrido-2-yl, pyrido-3-yl, and pyrido-4-yl), pyrimidyl (e.g., pyrimido-2-yl, and pyrimido-4-yl), triazinyl (e.g., 1,3,4-triazin-2-yl, and 1,3,5-triazin-4-yl), pyridazinyl (e.g., pyridazin-3-yl), and pyrazinyl. In some embodiments, the heterocyclyl group comprises a heterocyclyl ring fused to one or more (e.g., 1 or 2) different cyclic groups (e.g., carbocyclic rings, heterocyclic rings), where the groups or bonds are located on the heterocyclyl ring, and in some embodiments, the bonds are heteroatoms contained within the heterocyclyl ring.

[0021] As used herein, the term "heteroaryl," used alone or as part of a larger phrase (e.g., "heteroarylalkyl" (also called "heteroaralkyl") or "heteroarylalkoxy" (also called "heteroaralkoxy")), refers to a monocyclic, bicyclic, or tricyclic ring system having 5 to 12 ring atoms, where at least one ring is aromatic and contains at least one heteroatom. In one embodiment, the heteroaryl comprises a 5- to 6-membered monocyclic aromatic group in which one or more ring atoms are O, N, or S. Representative examples of heteroaryl groups include thienyl, furyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxadiazolyl, tetrazolyl, thiatriazolyl, oxatriazolyl, pyridyl, pyrimidyl, imidazopyridyl, pyrazinyl, pyridadinyl, triazinyl, tetradinyl, tetrazolo[1,5-b]pyridazinyl, prinyl, deazaprinyl, and benzoxazolyl. Examples include lyl, benzofuryl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, benzimidazolyl, indolyl, 1,3-thiazole-2-yl, 1,3,4-triazole-5-yl, 1,3-oxazole-2-yl, 1,3,4-oxadiazole-5-yl, 1,2,4-oxadiazole-5-yl, 1,3,4-thiadiazole-5-yl, 1H-tetrazole-5-yl, and 1,2,3-triazole-5-yl. In some embodiments, the heteroaryl group comprises a heteroaryl ring fused to one or more (e.g., one or two) different cyclic groups (e.g., a carbocyclic ring or a heterocyclic ring), where the group or bond site is on the heteroaryl ring, and in some embodiments, the bond site is a heteroatom contained in the heterocyclyl ring.

[0022] Unless otherwise specified, to the extent not further defined for any particular group in the compound of Formula I or II, any of the groups described in the present specification may be substituted or unsubstituted. To the extent not otherwise disclosed for any particular group, representative examples of substituents include alkyl (e.g., C1~C6, C1~C5, C1~C4, C1~C3, C1~C2, C1), substituted alkyl (e.g., substituted C1~C6, C1~C5, C1~C4, C1~C3, C1~C2, C1), alkoxy (e.g., C1~C6, C1~C5, C1~C4, C1~C3, C1~C2, C1), substituted alkoxy (e.g., substituted C1~C6, C1~C5, C1~C4, C1~C3, C1~C2, C1), haloalkyl (e.g., CF3), alkenyl (e.g., C2~C6, C2~C5, C2~C4, C2~C3, C2), substituted alkenyl (e.g., substituted C2~C6, C2~C5, C2~C4, C2~C3, C2), alkynyl (e.g., C2~C6, C2~C5, C2~C4, C2~C3, C2), substituted alkynyl (e.g., substituted C2~C6, C2~C5, C2~C4, C2~C3, C2), cyclic (e.g., C3~C 12 , C5~C6), substituted cyclic (e.g., substituted C3~C 12 , C5~C6), carbocyclic (e.g., C3~C 12 , C5~C6), substituted carbocyclic (e.g., substituted C3~C 12 , C5~C6), heterocyclyl (e.g., 3~12-membered, 5~6-membered), substituted heterocyclyl (e.g., substituted 3~12-membered, 5~6-membered), aryl (e.g., benzyl and phenyl), substituted aryl (e.g., substituted benzyl or substituted phenyl), heteroaryl (e.g., pyridyl or pyrimidyl), substituted heteroaryl (e.g., substituted pyridyl or substituted pyrimidyl), aralkyl (e.g., benzyl), substituted aralkyl (e.g., substituted benzyl), halo, hydroxyl, aryloxy (e.g., C6~C 12 , C6), substituted aryloxy (e.g., substituted C6~C 12 , C6), alkylthio (e.g., C1~C6), substituted alkylthio (e.g., substituted C1~C6), arylthio (e.g., C6~C 12 , C6), substituted arylthio (e.g., substituted C6~C 12Examples of substituents include C6, cyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amide, substituted amide, thio, substituted thio, sulfinyl, substituted sulfinyl, sulfonyl, substituted sulfonyl, sulfinamide, substituted sulfinamide, sulfonamide, substituted sulfonamide, urea, substituted urea, carbamate, substituted carbamate, amino acid, and peptide group. In some embodiments, substituents are halo (e.g., F, and Cl), alkylhalo (e.g., CF3, CHF2, CH2F, CCl3, CHCl2, CH2Cl), cyano, hydroxy, (C1-C6 alkyl, (C1-C6) alkoxy, hydroxy(C1-C6) alkyl, amino, amino(C1-C6) alkyl, amino-di(C1-C6) alkyl, or (C3-C7) cycloalkyl.

[0023] In one embodiment, the compounds of the present disclosure are represented by formula I or II:

[0024] [ka] or a pharmaceutically acceptable salt or stereoisomer thereof, During the ceremony, Deglon binds to cereblon (CRBN) or von Hippel-Lindau (VHL) tumor suppressors. The linker represents the part that covalently bonds the degron to the nitrogen atom.

[0025] In some embodiments, the degron bound to cereblon is represented by one of the following structures:

[0026] [ka] During the ceremony, X is CH2 or C(O), R1 is absent, CH2,

[0027] [ka] A 5-11 member monocyclic or bicyclic heterocycline containing one or two heteroatoms independently selected from NH, O, or N, O, and S, X' is either nonexistent or C(O), Y is either N or CH. R2 is an optional substituent.

[0028] In some embodiments, the degron is of formula D1:

[0029] [ka]

[0030] In some embodiments, X is C(O). In some embodiments, X is CH2.

[0031] In some embodiments, R1 is absent, CH2,

[0032] [ka] It is either NH or O.

[0033] The disclosed compounds also encompass all combinations of the specific X and R1 groups disclosed above.

[0034] In some embodiments, the degron is of formula D2:

[0035] [ka]

[0036] In some embodiments, X' is absent.

[0037] In some embodiments, R2 is a halo (e.g., F, and Cl), an alkylhalo (e.g., CF3, CHF2, CH2F, CCl3, CHCl2, CH2Cl), a cyano, a hydroxyl, a (C1-C6)alkyl, a (C1-C6)alkoxy, a hydroxy(C1-C6)alkyl, an amino, an amino(C1-C6 alkyl, amino-di(C1-C6)alkyl, and a (C3-C7)cycloalkyl. In some embodiments, R2 is a fluoro.

[0038] In some embodiments, R1 is absent, CH2,

[0039] [ka] It is either NH or O.

[0040] In some embodiments, R1 is a six-membered monocyclic heterocycline containing one or two heteroatoms independently selected from N, O, and S. In some embodiments, R1 is piperazinyl. In some embodiments, R1 is piperidinyl.

[0041] In some embodiments, R1 is a nine-membered bicyclic heterocycline containing one or two heteroatoms independently selected from N, O, and S. In some embodiments, R1 is 2,7-diazaspiro[3.5]nonane

[0042] [ka] That is the case.

[0043] The disclosed compounds also encompass all combinations of the specific X', R1, and R2 groups disclosed above.

[0044] In some embodiments, the degron is of formula D3:

[0045] [ka]

[0046] In some embodiments, R1 is CH2,

[0047] [ka] It is either NH or O.

[0048] In some embodiments, the degron is of formula D4:

[0049] [ka]

[0050] In some embodiments, Y is CH.

[0051] In some embodiments, R2 is a halo (e.g., F or Cl), an alkylhalo (e.g., CF3, CHF2, CH2F, CCl3, CHCl2, CH2Cl), a cyano, a hydroxyl, a (C1-C6) alkyl, a (C1-C6) alkoxy, a hydroxy(C1-C6) alkyl, an amino, an amino(C1-C6 alkyl, amino-di(C1-C6 alkyl, and (C3-C7) cycloalkyl). In some embodiments, R2 is a methoxy.

[0052] In some embodiments, R1 is absent, CH2,

[0053] [ka] It is either NH or O.

[0054] The disclosed compounds also encompass all combinations of the specific Y, R1, and R2 groups disclosed above.

[0055] In some embodiments, the degron is of formula D5:

[0056] [ka]

[0057] In some embodiments, X is C(O).

[0058] In some embodiments, the degron is represented by one of the following structures:

[0059] [ka]

[0060] [ka]

[0061] In some embodiments, the degron coupled to VHL is represented by D6:

[0062] [ka] or its stereoisomer, During the ceremony, R3 is hydrogen, or an optionally substituted C1-C3 alkyl group, or R3 and R4, together with the carbon atoms to which they are bonded, form a cyclopropyl group. R4 is hydrogen, methyl, or

[0063] [ka] And, R5 is C(O)CR6R7R8,

[0064] [ka] And, R6 and R7 are either hydrogen or R6 and R7, together with the carbon atoms to which they are bonded, form a cyclopropyl group. R8 is hydrogen, fluoro, cyano, or NMe2. Y is hydrogen,

[0065] [ka] And, During the ceremony,

[0066] [ka] This is a bond between a degron and a linker, however, there is only one bond between a degron and a linker.

[0067] In some embodiments, R3 and R4 are hydrogen.

[0068] In some embodiments, R3 is hydrogen and R4 is methyl.

[0069] In some embodiments, R3 is an optionally substituted C1-C3 alkyl group, and R4 is hydrogen.

[0070] In some embodiments, R3 is an optionally substituted C1-C3 alkyl group, and R4 is methyl.

[0071] In some embodiments, R3 and R4, together with the carbon atoms to which they are bonded, form a cyclopropyl group.

[0072] In some embodiments, Y is hydrogen.

[0073] In some embodiments, Y is

[0074] [ka] That is the case.

[0075] In some embodiments, R5 is

[0076] [ka] That is the case.

[0077] In some embodiments, R5 is C(O)CR6R7R8.

[0078] In some embodiments, R6, R7, and R8 are hydrogen.

[0079] In some embodiments, R6 and R7, together with the carbon atoms to which they are bonded, form a cyclopropyl group, and R8 is hydrogen, fluoro, cyano, or NMe2.

[0080] In some embodiments, R3 is hydrogen, and R4 is

[0081] [ka] And Y is hydrogen.

[0082] In some embodiments, R3 is hydrogen, and R4 is

[0083] [ka] Y is hydrogen, R5 is C(O)CR6R7R8, where R6 and R7, together with the carbon atoms to which they are bonded, form a cyclopropyl group, and R8 is fluoro, cyano, or NMe2.

[0084] In some embodiments, Y is

[0085] [ka] R3 and R4 are hydrogen.

[0086] In some embodiments, Y is

[0087] [ka] R3 and R4 are hydrogen, R5 is C(O)CR6R7R8, where R6 and R7, together with the carbon atoms to which they are bonded, form a cyclopropyl group, and R8 is fluoro, cyano, or NMe2.

[0088] In some embodiments, R5 is

[0089] [ka] Y is hydrogen, and R3 and R4 are hydrogen.

[0090] In some embodiments, R5 is

[0091] [ka] Y is hydrogen, R3 is hydrogen, and R4 is methyl.

[0092] In some embodiments, R5 is

[0093] [ka] Y is hydrogen, R3 is an optionally substituted C1-C3 alkyl group, and R4 is hydrogen.

[0094] In some embodiments, R5 is

[0095] [ka] Here, Y is hydrogen, R3 is an optionally substituted C1-C3 alkyl, and R4 is methyl.

[0096] In some embodiments, the degron bound to the VHL is represented by one of the following structures:

[0097] [ka] In the formula, Y is O, NH, CH2, or a stereoisomer thereof.

[0098] In formula I or II, the linker ("L") represents the part that covalently bonds the degron to the nitrogen atom.

[0099] In some embodiments, the linker comprises at least one alkylene group, and in some embodiments, an alkylene chain (e.g., having 1 to 20 alkylene units) or a divalent alkylene chain, any of which is -O-, -S-, -N(R')-,

[0100] [ka] -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(NOR')-, -C(O)N(R')-, -C(O)N(R')C(O)-, -C(O)N(R')C(O)N(R')- , -N(R')C(O)-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -OC(O)N(R')-, -C(NR')-, -N(R')C(NR')-, -C(NR')N(R' )-, -N(R')C(NR')N(R')-, -OB(Me)O-, -S(O)2-, -OS(O)-, -S(O)O-, -S(O)-, -OS(O)2-, -S(O)2O-, -N(R')S(O )2-, -S(O)2N(R')-, -N(R')S(O)-, -S(O)N(R')-, -N(R')S(O)2N(R')-, -N(R')S(O)N(R')-, optionally substituted C3~C 12 The molecule may be interrupted by at least one of the following: a carbocyclene, an optionally substituted 3- to 12-membered heterocyclene, an optionally substituted 5- to 12-membered heteroarylene, or any combination thereof, and / or terminated at either or both ends by at least one thereof, where R' is H or a C1- to C6 alkyl group, and the interrupting group and the one or both terminal groups may be the same or different.

[0101] "Carbocyclene" refers to a divalent carbon ring group, which can be optionally substituted.

[0102] "Heterocyclene" refers to a divalent heterocyclyl group, which can be optionally substituted.

[0103] "Heteroarylene" refers to a divalent heteroaryl group, which can be optionally substituted.

[0104] In some embodiments, the linker comprises an alkylene chain having 3 to 11 alkylene units and terminated with C(O). In some embodiments, the linker comprises an alkylene chain having 4 to 11 alkylene units. In some embodiments, the linker comprises an alkylene chain having 3 to 8 alkylene units.

[0105] In some embodiments, the linker is -O-, -S-, -N(R')-,

[0106] [ka] -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(NOR')-, -C(O)N(R')-, -C(O)N(R')C(O)-, -C(O)N(R')C(O)N(R')- , -N(R')C(O)-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -OC(O)N(R')-, -C(NR')-, -N(R')C(NR')-, -C(NR')N(R' )-, -N(R')C(NR')N(R')-, -OB(Me)O-, -S(O)2-, -OS(O)-, -S(O)O-, -S(O)-, -OS(O)2-, -S(O)2O-, -N(R')S(O )2-, -S(O)2N(R')-, -N(R')S(O)-, -S(O)N(R')-, -N(R')S(O)2N(R')-, -N(R')S(O)N(R')-, optionally substituted C3~C 12 A polyethylene glycol (PEG) chain that may be interrupted by at least one of carbosislene, optionally substituted 3-12 member heterocyclene, optionally substituted 5-12 member heteroarylene, or any combination thereof, and / or terminated at either or both ends by at least one thereof, wherein R' is H or C1-C6 alkyl, and the interrupting group and one or both terminal groups may be the same or different.

[0107] In some embodiments, the linker comprises a polyethylene glycol chain having 1 to 5 PEG units and terminated with C(O).

[0108] Typical examples of linkers containing polyethylene glycol chains include the following:

[0109] [ka] In the formula, n is an integer between 2 and 10, and examples include the following:

[0110] [ka]

[0111] In some embodiments, the linker is of formula L0:

[0112] [ka] or its stereoisomer, in the formula, p1 is an integer selected from 0 to 6. p2 is an integer selected from 0 to 12. p3 is an integer selected from 1 to 12. Each W is independent, absent, CH2, O, S, NR 10 , or C(O)NR 10 And, Each R 10 These are independently hydrogen or C1-C6 alkyl groups. W1 and W2 are independently absent, (CH2) 1~3 , O, or NH, Z1 and Z2 are independently: nonexistent, -O-, -S-, -N(R 10 )-,

[0113] [ka] -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(NOR 10 )-,-C(O)N(R 10 )-,-C(O)N(R 10 )C(O)-, -C(O)N(R 10 )C(O)N(R10 )-, -N(R 10 )C(O)-, -N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)O-, -OC(O)N(R 10 )-, -C(NR 10 )-, -N(R 10 )C(NR 10 )-, -C(NR 10 )N(R 10 )-, -N(R 10 )C(NR 10 )N(R 10 )-, -OB(Me)O-, -S(O)2-, -OS(O)-, -S(O)O-, -S(O)-, -OS(O)2-, -S(O)2O-, -N(R 10 )S(O)2-, -S(O)2N(R 10 )-, -N(R 10 )S(O)-, -S(O)N(R 10 )-, -N(R 10 )S(O)2N(R 10 )-, -N(R 10 )S(O)N(R 10 )-, optionally substituted C3~C 12 carbocyclene, optionally substituted 3~12 membered heterocyclene, or optionally substituted 5~12 membered heteroarylene, the linker is covalently bonded to the degron via the position adjacent to W2 of the degron

[0114]

Chemical formula

[0115]

Chemical formula

[0116]

Chemical formula

[0117] [ka] It forms a covalent bond with the nitrogen atom via this.

[0118] In some embodiments, p1 is an integer selected from 0 to 3.

[0119] In some embodiments, p2 is an integer selected from 0 to 10. In some embodiments, p2 is an integer selected from 0 to 6. In some embodiments, p2 is an integer selected from 0 to 3.

[0120] In some embodiments, p3 is an integer selected from 1 to 10. In some embodiments, p3 is an integer selected from 1 to 6.

[0121] In some embodiments, each W is independently absent, O, or C(O)NR 10 That is the case.

[0122] In some embodiments, W1 and W2 are absent.

[0123] In some embodiments, Z1 and Z2 are independently absent, -O-, -N(R 10 )-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)N(R 10 )-,-N(R 10 The C(O)-, optionally substituted C6-carbocylene, optionally substituted 5- to 6-membered heterocyclene, or optionally substituted 5- to 6-membered heteroarylene.

[0124] In some embodiments, formula L0 is one of formulas L0a to L0g:

[0125]

Chemical formula

[0126] In some embodiments, the linker is as follows:

[0127]

Chemical formula

[0128]

Chemical formula

[0129] In some embodiments,

[0130]

Chemical formula

[0131]

Chemical formula

[0132] In some embodiments, the compound of Formula I is represented by any one of the following structures:

[0133]

Chemical formula

[0134]

Chemical formula

[0135]

Chemical formula

[0136] [ka]

[0137] [ka]

[0138] [ka] Or a pharmaceutically acceptable salt or stereoisomer thereof.

[0139] In some embodiments, the compound of formula II is represented by one of the following structures:

[0140] [ka]

[0141] [ka] Or a pharmaceutically acceptable salt or stereoisomer thereof.

[0142] In some embodiments, the compounds disclosed herein include:

[0143] [ka]

[0144] The figure refers to the following compound:

[0145] [ka]

[0146] [ka]

[0147] The compounds of this disclosure may be in the form of free acids, free bases, or pharmaceutically acceptable salts. A pharmaceutically acceptable salt of a compound of this disclosure may be formed, for example, by the reaction of a suitable free base of the compound of this invention with a suitable pharmaceutically acceptable acid in a suitable solvent under standard conditions well known in the art. See, for example, Gould, PL, "Salt selection for basic drugs," International Journal of Pharmaceutics, 33:201-217 (1986); Bastin, RJ, et al., "Salt Selection and Optimization Procedures for Pharmaceutical New Chemical Entities," Organic Process Research and Development, 4:427-435 (2000); and Berge, SM, et al., "Pharmaceutical Salts," Journal of Pharmaceutical Sciences, 66:1-19 (1977).

[0148] The compounds of this disclosure may have at least one chiral center and therefore may exist in the form of stereoisomers, which, as used herein, encompass all isomers of individual compounds that differ only in the orientation of their atoms in space. The term stereoisomer includes enantiomers (enantiomers of a compound, including the (R-) or (S-) configuration), mixtures of enantiomers of a compound (physical mixtures of enantiomers, and racemates or racemic mixtures), geometric (cis / trans, or E / Z, R / S) isomers of a compound, and isomers of a compound (diastereomers) that have one or more chiral centers that are not mirror images of each other. The chiral centers of a compound can undergo epimerization in vivo; therefore, for these compounds, administration of the compound in its (R-) form is considered equivalent to administration of the compound in its (S-) form. Accordingly, the compounds of this disclosure can be prepared and used in the form of individual isomers and substantially without other isomers, or in the form of mixtures of various isomers, for example, racemic mixtures of stereoisomers.

[0149] In some embodiments, the compound of formula I or II is an isotopic derivative in that it has at least one desired isotopic substitution of an atom in an amount exceeding the natural abundance of the isotope, i.e., an enriched amount. In one embodiment, the compound contains deuterium or a plurality of deuterium atoms. As used herein, the term "hydrogen," i.e., H, means protium ( 1 H), and deuterium 2 This refers to all isotopes of hydrogen, including H). As used herein, the term “compound” includes isotopic derivatives.

[0150] The compounds of this disclosure may also be in the form of N-oxides, crystalline forms (also known as polymorphs), cocrystals, active metabolites of compounds having the same type of activity, prodrugs, tautomers, and non-solvated and solvated (e.g., hydrated) forms of the compounds with pharmaceutically acceptable solvents such as water and ethanol. As used herein, the term “compound” encompasses all of these forms.

[0151] Synthesis method In some embodiments, this disclosure relates to methods for preparing compounds of formula I or II, or pharmaceutically acceptable salts or stereoisomers thereof. Broadly speaking, the compounds, or pharmaceutically acceptable salts or stereoisomers thereof, may be prepared by any method known to be applicable to the preparation of chemically related compounds. The compounds of this disclosure will be better understood in relation to the synthetic schemes described in various examples illustrating non-limiting methods by which the compounds of this disclosure may be prepared.

[0152] Pharmaceutical composition Another aspect of this disclosure relates to a pharmaceutical composition comprising a compound of formula I or II, or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier. The term “pharmaceutically acceptable carrier” means, as known in the art, a pharmaceutically acceptable material, composition, or vehicle suitable for administering the compounds of this disclosure to a mammal. Suitable carriers may include, for example, liquids (both aqueous and non-aqueous, and combinations thereof), solids, encapsulating materials, gases, and combinations thereof (e.g., semi-solids), and gases that function to transport or deliver the compound from one organ or part of body to another organ or part of body. The carrier is “acceptable” in the sense that it is physiologically inert to the other components of the formulation, compatible with the other components of the formulation, and not harmful to the subject or patient. Depending on the type of formulation, the composition may also include one or more pharmaceutically acceptable excipients.

[0153] More broadly, compounds of formula I or II, as well as their pharmaceutically acceptable salts and stereoisomers, can be formulated into a given type of composition according to conventional pharmaceutical practices such as conventional mixing, dissolution, granulation, sugar-coated tablet preparation, polishing, emulsification, encapsulation, encapsulation, and compression processes (e.g., Remington: The Science and Practice of Pharmacy (20)). thSee eds., ARGennaro, Lippincott Williams & Wilkins, 2000 and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and JCBoylan, 1988–1999, Marcel Dekker, New York (each of these is incorporated herein by reference in whole). The type of formulation depends on the mode of administration, which may include enteral (e.g., oral, oral, sublingual, and rectal), parenteral (e.g., subcutaneous (sc), intravenous (iv), intramuscular (im), and intrasternal injection or infusion techniques, intraocular, intra-arterial, intramedullary, intrathecal, intraventricular, percutaneous, intradermal, vaginal, intraperitoneal, mucosal, nasal, intratracheal infusion, bronchial infusion, and inhalation), and topical (e.g., percutaneous).

[0154] Generally, the most appropriate route of administration depends on various factors, including, for example, the properties of the drug (e.g., its stability in the gastrointestinal environment) and / or the condition of the subject (e.g., whether the subject can tolerate oral administration). For example, parenteral administration (e.g., intravenous) may also be advantageous in that it allows for relatively rapid administration of the compound, such as in single-dose treatments and / or in cases of acute conditions.

[0155] In some embodiments, the compound is formulated for intravenous administration (e.g., systemic intravenous injection).

[0156] Injectable preparations for parenteral administration may include sterile aqueous solutions or oily suspensions. They can be formulated according to standard techniques using appropriate dispersants or wetting agents and suspending agents. Sterile injectable preparations may be sterile injectable solutions, suspensions, or emulsions in non-toxic, parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable vehicles and solvents that may be used (included in the term "carrier" together with diluents) include water, Ringer's solution, USP, and isotonic sodium chloride solutions. Furthermore, sterile non-volatile oils have conventionally been used as solvents or suspension media. For this purpose, any non-irritating non-volatile oil, including synthetic mono or diglycerides, may be used. In addition, fatty acids such as oleic acid are used in the preparation of injectables. Injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable medium before use. The effect of a compound can be extended by delaying its absorption, which can be achieved by using a liquid suspension or a less water-soluble crystalline or amorphous material. Longer absorption of a compound from parenterally administered formulations can also be achieved by suspending the compound in an oily vehicle.

[0157] In some embodiments, compounds of formula I or II may be administered topically rather than systemically, for example, by direct injection of a conjugate into an organ, often in the form of a depot preparation or a sustained-release formulation. In certain embodiments, long-acting formulations are administered by implantation (e.g., subcutaneous or intramuscular) or by intramuscular injection. Injectable depot formulations are prepared by forming a microcapsule matrix of the compound with biodegradable polymers, such as polylactide-polyglycolide, poly(orthoester), and poly(in anhydrous). The release rate of the compound can be controlled by changing the ratio of the compound to the polymer and the properties of the particular polymer used. Depot injection formulations are also prepared by encapsulating the compound in liposomes or microemulsions compatible with body tissues. Furthermore, in other embodiments, the compound is delivered by a targeted drug delivery system, for example, liposomes coated with organ-specific antibodies. In such embodiments, the liposomes are targeted to an organ and selectively taken up by the organ.

[0158] The composition may contain excipients, most commonly known as preservatives, antioxidants, buffers, solubilizers, and surfactants. Typical examples of suitable preservatives include alcohols, quaternary amines, organic acids, parabens, and phenols. Typical examples of suitable antioxidants include ascorbic acid and its esters, sodium bisulfite, butylated hydroxytoluene, butylated hydroxyanisole, tocopherol, and chelating agents such as EDTA and citric acid. Typical examples of suitable buffers include citric acid, hydrochloric acid, and lactate buffer. Typical examples of suitable solubilizers include quaternary ammonium chloride, cyclodextrin, benzyl benzoate, lecithin, and polysorbate.

[0159] In other embodiments, the composition may be formulated for oral, sublingual, sublingual (examples include tablets, lozenges, and gels), inhalation, topical, ocular, and rectal administration.

[0160] Dosage The amount of compound I or II administered to a cancer patient may be effective in producing a desired therapeutic response in the patient. Therefore, the amount of compound I or II, or its pharmaceutically acceptable salt or stereoisomer, may induce a positive modification in the treated disease or disorder, prevent the onset or progression of the disease or disorder, alleviate one or more symptoms of the treated disease or disorder in the subject, kill or inhibit the proliferation of cancer cells, or reduce the amount of BPTF in cancer cells.

[0161] The total daily dose of a compound and its use may be determined, for example, by the attending physician using sound medical judgment and in accordance with standard medical practice. The specific dose for any particular subject may depend on a variety of factors, including the disease or disorder being treated and its severity (e.g., its current condition), the subject's age, weight, overall health, sex, and diet, the timing of administration, route of administration, and excretion rate of the specific compound used, the duration of treatment, any drugs used in combination with or concurrently with the compound, and similar factors well known in the medical field (e.g., Goodman and Gilma's *The Pharmacological Basis of Therapeutics*, 1). 0t See h Edition, A. Gilman, J. Hardman and L. Limbird, eds., McGraw-Hill Press, 155–173, 2001 (which is incorporated herein by reference in its entirety).

[0162] The compounds of formulas I and II, as well as their pharmaceutically acceptable salts and stereoisomers, may be effective over a wide dose range. In some embodiments, the total daily dose (e.g., for adults) may range from about 0.001 to about 1600 mg, 0.01 to about 1600 mg, 0.01 to about 500 mg, about 0.01 to about 100 mg, about 0.5 to about 100 mg, about 100 to 400 mg per day, about 1 to about 50 mg per day, and about 5 to about 40 mg per day, and in yet other embodiments, about 10 to about 30 mg per day. Individual doses may be formulated to contain the desired dose depending on the number of times the compound is administered per day. For example, capsules may be formulated using approximately 1 to approximately 200 mg of the compound (e.g., 1, 2, 2.5, 3, 4, 5, 10, 15, 20, 25, 50, 100, 150, and 200 mg). In some embodiments, individual doses may be formulated to contain a desired dose depending on the number of times the compound is administered per day.

[0163] How to use In some embodiments, the Disclosure relates to methods for treating cancer by reducing the level or activity of BPTF. The methods include administering a compound of formula I or II, or a pharmaceutically acceptable salt or stereoisomer thereof, to a subject of interest.

[0164] Cancer is associated with (e.g., characterized by or mediated by) abnormal BPTF activity (e.g., elevated levels of BPTF, or otherwise functionally abnormal BPTF, e.g., mutant BPTF activity) compared to non-pathological conditions.

[0165] As used herein, the term “Subject” (or “Patient”) includes all members of the animal kingdom who are susceptible to or have a disease or disorder described herein. In some embodiments, a subject is a mammal, e.g., a human, or a non-human mammal. A subject who “needs” treatment according to this disclosure may have a particular disease or disorder, be “suffering from or suspected of having,” be diagnosed positive for, or otherwise present with a sufficient number of risk factors, or a sufficient number or combination of signs or symptoms, that would allow a healthcare professional to diagnose or suspect that the subject has a disease or disorder. Thus, subjects who have a particular disease or disorder and those suspected of having a particular disease or disorder are not necessarily two separate groups.

[0166] Cancers that may be suitable for treatment with the disclosed compounds include carcinomas, sarcomas, and blood-borne (hematological) cancers. In some embodiments, cancers include melanoma, endometrial cancer, urothelial carcinoma of the bladder, invasive breast cancer, undifferentiated gastric adenocarcinoma, esophagogastric junction adenocarcinoma, cervical squamous cell carcinoma, non-small cell lung cancer, pleural mesothelioma, hepatocellular carcinoma, colorectal adenocarcinoma, pancreatic adenocarcinoma, ovarian epithelial tumors, sarcomas, ocular melanoma, non-testicular germ cell tumors, adrenocortical carcinoma, head and neck squamous cell carcinoma, mature B-cell neoplasms, pheochromocytoma, diffuse glioblastoma, renal non-clear cell carcinoma, thymic epithelial tumors, well-differentiated thyroid cancer, esophageal squamous cell carcinoma, glioblastoma, and pre- These include prostate adenocarcinoma, leukemia, clear cell carcinoma of the kidney, other neuroepithelial tumors, cervical adenocarcinoma, testicular tumors, cholangiocarcinoma, acute myeloid leukemia, low-grade glioblastoma of the brain, diffuse large B-cell lymphoma, glioblastoma multiforme, renal pigment neoplasm, clear cell carcinoma of the kidney, papillary cell carcinoma of the kidney, hepatocellular carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, ovarian serous cystadenocarcinoma, pancreatic adenocarcinoma, pheochromocytoma, paraganglioma, cutaneous melanoma, gastric adenocarcinoma, testicular germ cell tumor, thymoma, thyroid cancer, uterine sarcoma, endometrial cancer, or uveal melanoma.

[0167] Representative examples of sarcomas that can be treated in accordance with this disclosure include fibrosarcoma, malignant fibrous histiocytoma, dermatofibrosarcoma, liposarcoma, leiomyosarcoma, angiosarcoma, Kaposi's sarcoma, lymphangiosarcoma, synovial sarcoma, malignant peripheral nerve sheath tumor, neurofibrosarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, germ cell and alveolar soft tissue sarcoma, infantile angiopericytoma, extraskeletal myxoid chondrosarcoma, and extraskeletal mesenchymal sarcoma.

[0168] In some embodiments, the patient has rhabdoid sarcoma. In some embodiments, rhabdoid sarcoma affects the kidneys and other soft tissues. Also known as rhabdoid tumor, or malignant rhabdoid tumor (MRT), this is a rare tumor that afflicts children. In some embodiments, rhabdoid sarcoma grows in the central nervous system or brain and is also known as atypical teratomatous rhabdoid tumor (AT / RT).

[0169] In some embodiments, the sarcoma is a rhabdomyosarcoma.

[0170] These and other aspects of the present disclosure will be further understood by considering the following embodiments. These embodiments are intended to illustrate certain embodiments of the present disclosure, but are not intended to limit the scope defined by the claims. [Examples]

[0171] Example 1: Synthesis of 4-((4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)-4-oxobutyl)amino)-2-(2,6-dioxopiperidine-3-yl)isoindorin-1,3-dione (dBPTF-1, NVS-BPTF-1A)

[0172] [ka]

[0173] 7-Bromo-3-methyl-2H-pyrido[1,2-a]pyrimidine-2,4(3H)-dione To a solution of 5-bromopyridine-2-amine (20 g, 114.93 mmol) in diethyl 2-methyl malonate (28 mL), concentrated HCl (20 drops) was added, and the reaction mixture was stirred under nitrogen at 145°C for 110 hours. The reaction mixture was diluted with toluene (100 mL), filtered, and the solid was washed with toluene (20 mL). The filtrate was concentrated to obtain the title compound as a black solid (3.8 g, 10%). 1 H NMR (400MHz, DMSO) δ8.94(d,J=1.8Hz,1H),8.08(dd,J=9.3,2.2Hz,1H),7.35(dd,J=9.3,0.5Hz,1H),1.92(s,3H). LCMS:[M+H] + = 254.9.

[0174] 7-Bromo-2-chloro-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one A solution of 7-bromo-3-methyl-2H-pyrido[1,2-a]pyrimidine-2,4(3H)-dione (1 g, 5.78 mmol) in POCl3 (2 mL) was heated to 120 °C and stirred under nitrogen for 16 hours. The reaction mixture was concentrated under reduced pressure, and the residue was diluted with water (30 mL) and extracted with toluene (50 mL x 6). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under reduced pressure to obtain the residue. This residue was purified by silica gel chromatography using PE:toluene = 5:1 elution to obtain the title compound as a yellow solid (0.85 g, 90%). LCMS:[M+H] + =275.0.

[0175] 2-Chloro-7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one To a solution of 7-bromo-2-chloro-3-methyl-4H-pyridino[1,2-a]pyrimidine-4-one (200 mg, 0.73 mmol), KOAc (150 mg, 1.47 mmol), and 1-cyclopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (172 mg, 0.73 mmol) in 1,4-dioxane (2 mL) and H2O (0.4 mL), Pd(dppf)Cl2 (54 mg, 0.073 mmol) was added, and the reaction mixture was stirred under nitrogen at 100°C for 2 hours. The reaction mixture was poured into water (15 mL) and extracted with ethylethanol (10 mL x 3). The combined organic layers were washed with brine (20 mL x 2), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. This residue was purified by silica gel chromatography using PE:siRNA=3:2 elution to obtain the title compound as a yellow oily substance (0.11 g, 82%). 1 H NMR(400MHz,DMSO)δ9.03(s,1H),8.55(s,1H),8.33-8.21(m,1H),8.08(s,1H),7.69(d,J=9. 2Hz,1H),3.84-3.74(m,1H),2.23(s,3H),1.16-1.06(m,2H),1.05-0.96(m,2H).LCMS:[M+H] + = 300.9.

[0176] tert-butyl 4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-carboxylate In toluene (0.5 mL), a solution of 2-chloro-7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4H-pyridino[1,2-a]pyrimidine-4-one (20 mg, 0.063 mmol), Cs2CO3 (44 mg, 0.126 mmol), and tert-butyl 4-((4-amino-3-fluorophenyl)sulfonyl)piperazine-1-carboxylate (30 mg, 0.082 mmol) was added. Pd2(dba)3 (6 mg, 0.0063 mmol) and BINAP (8 mg, 0.0126 mmol) were added to the reaction mixture. The reaction mixture was degassed three times with nitrogen and stirred at 110°C for 2 hours. The reaction mixture was concentrated, diluted with water (10 mL), and extracted with ELISA (10 mL x 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. This residue was purified by silica gel chromatography using PE:siRNA=1:1.2 elution to obtain the title compound as a white solid (0.04 g, 92%). 1 H NMR(400MHz,DMSO)δ9.02(d,J=1.7Hz,1H),8.47(d,J=3.0Hz,2H),8.12-7.98(m,3H),7.61(d,J=10.3Hz,1H),7.56(d,J=8.5Hz,1H),7.41(d,J=9 .2Hz,1H),3.84-3.74(m,1H),3.45-3.40(m,4H),2.98-2.91(m,4H),2.1 8(s,3H),1.35(s,9H),1.13-1.07(m,2H),1.03-0.96(m,2H).LCMS:[M+H] + = 624.3.

[0177] 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one A solution of tert-butyl 4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyridino[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-carboxylate (40 mg, 0.064 mmol) in HCl / 1,4-dioxane (4 M, 0.5 mL) was stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a yellow solid (40 mg, crude), which was used without further purification.

[0178] 4-((2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-4-yl)aminobutanoic acid In DMSO (5 mL), a solution of 2-(2,6-dioxopiperidine-3-yl)-4-fluoroisoindoline-1,3-dione (500 mg, 1.81 mmol) was added to 4-aminobutyric acid (560 mg, 5.43 mmol) and DIEA (935 mg, 7.24 mmol). The reaction mixture was stirred at 120°C for 16 hours. The reaction mixture was poured into water (50 mL) and extracted with RINKAN (80 mL x 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the title compound as a yellow oil (800 mg, crude). This was used without further purification. LCMS: [M+H] + =360.0.

[0179] Perfluorophenyl 4-((2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-4-yl)aminobutanoate In 10 mL of DMF, a solution of 4-((2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-4-yl)aminobutyric acid (800 mg, crude, 2.23 mmol), 2,3,4,5,6-pentafluorophenol (492 mg, 2.68 mmol), and DCC (688 mg, 3.45 mmol) was stirred at room temperature for 16 hours. The reaction mixture was poured into 100 mL of water and extracted with RINKAN (80 mL x 3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. This residue was purified by flash chromatography on a silica gel column (0%~30% EA in PE) to obtain the title compound as a yellow solid (70 mg, 7% in 2 steps). LCMS: [M+H] + = 526.3.

[0180] 4-((4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)-4-oxobutyl)amino)-2-(2,6-dioxopiperidine-3-yl)isoindorin-1,3-dione In DMSO (4 mL), 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (40 mg, 0.0765 mmol) and perfluorophenyl 4-((2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-4-yl)amino)butanoate (70 mg, 0.13 mmol) were mixed with DIEA (99 mg, 0.775 mmol), and the reaction mixture was stirred at 25°C for 2 hours. The reaction mixture was diluted with water (40 mL), filtered, and the solid was purified by preparative HPLC to obtain the title compound as a yellow solid (29 mg, 44%). 1H NMR(400MHz,DMSO)δ11.07(s,1H),9.01(d,J=1.7Hz,1H),8.46(s,2H),8.13-7.97(m,3H),7.64-7.59(m,1H),7.58-7 .48(m,2H),7.40(d,J=9.1Hz,1H),7.10(d,J=8.6Hz,1H),6.97(d,J=7.0Hz,1H),6.62(t,J=6.1Hz,1H),5.06-4.98(m ,1H),3.80-3.71(m,1H),3.62-3.50(m,4H),3.28-3.24(m,2H),3.00-2.91(m,4H),2.90-2.82(m,1H),2.62-2.51(m, 2H), 2.37(t,J=6.8Hz,2H),2.18(s,3H),2.05-1.96(m,1H),1.80-1.69(m,2H),1.12-1.07(m,2H),1.03-0.97(m,2H). LCMS:[M+H] + = 865.4.

[0181] Example 2: Synthesis of 4-((4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)-4-oxobutyl)amino)-2-(2,6-dioxopiperidine-3-yl)isoindorin-1,3-dione (dBPTF-2, NVS-BPTF-1B)

[0182] [ka]

[0183] tert-butyl 4-(4-hydroxybutyl)piperazine-1-carboxylate In a solution of tert-butylpiperazine-1-carboxylate (5 g, 26.84 mmol) and 4-bromobutan-1-ol (6.2 g) in ACN (60 mL), DIEA (10 mL) was gradually added while stirring at 100°C under nitrogen gas for 4 hours. The reaction mixture was concentrated, water (100 mL) was added, and it was extracted with EA (50 mL x 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. This residue was purified by silica gel chromatography using DCM:MeOH=8% elution to obtain the title compound as a colorless oil (2.1 g, 95%).

[0184] tert-butyl 4-(4-oxobutyl)piperazine-1-carboxylate In 5 mL of DCM, a solution of oxalyl chloride (10.66 mL, 7.74 mmol) was added dropwise to DMSO (1.1 mL) under nitrogen at -78°C for 0.5 hours with stirring. Then, in 5 mL of DCM, a solution of tert-butyl 4-(4-hydroxybutyl)piperazine-1-carboxylate (0.5 g, 1.94 mmol) was added dropwise at -78°C for 0.5 hours. TEA (2.7 mL, 19.35 mmol) was slowly added to the reaction mixture, and the reaction mixture was stirred at 25°C for 0.5 hours. The reaction mixture was poured into saturated sodium carbonate aqueous solution and extracted with DCM (30 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. This residue was purified by silica gel chromatography using DCM:MeOH=25:1 elution to obtain the title compound as a yellow oily substance (0.12 g, 90%). LCMS:[M+H] + = 257.2.

[0185] tert-butyl 4-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)piperazine-1-carboxylate In 4 mL of DCM, 100 mg, 0.19 mmol, 100 mg, 0.38 The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. This residue was purified by silica gel chromatography using DCM:MeOH=20:1 elution to obtain the title compound as a white solid (0.15 g, 100%). LCMS:[M+H] + = 764.4.

[0186] 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-((4-(4-(piperazin-1-yl)butyl)piperazin-1-yl)sulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one A solution of tert-butyl 4-(4-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)piperazine-1-carboxylate (0.13 g) was stirred in HCl / dioxane (2 mL) for 2 hours at 25°C. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a yellow solid (0.13 g, 85%).

[0187] 4-((4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)-4-oxobutyl)amino)-2-(2,6-dioxopiperidine-3-yl)isoindorin-1,3-dione In DMSO (2 mL), 129 mg of DIEA (1 mmol) was added to a solution of 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-((4-(4-(piperazin-1-yl)butyl)piperazin-1-yl)sulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (130 mg, 0.2 mmol) and 2-(2,6-dioxopiperidine-3-yl)-5-fluoroisoindorin-1,3-dione (67 mg, 0.24 mmol). The reaction mixture was stirred under nitrogen at 25°C, and then under microwave at 120°C for 1 hour. The reaction product was purified by preparative HPLC to obtain the title compound as a yellow solid (37 mg). 1 H NMR(400MHz,DMSO)δ11.07(s,1H),9.00(d,J=1.7Hz,1H),8.47(d,J=7.6Hz,2H),8.08(dd,J=9.2,2.0Hz,1H),8.04-7.9 8(m,2H),7.63(t,J=10.0Hz,2H),7.56(d,J=8.6Hz,1H),7.41(d,J=9.2Hz,1H),7.28(s,1H),7.20(d,J=8.7Hz,1H),5.06 (dd,J=12.8,5.5Hz,1H),3.77(s,1H),3.38(d,J=5.1Hz,4H),2.96(s,4H),2.91-2.83(m,1H),2.55(d,J=9.2Hz,2H),2.4 4(s,8H),2.28(d,J=5.8Hz,4H),2.18(s,3H),2.05-1.97(m,1H),1.41(s,4H),1.09(d,J=3.6Hz,2H),1.03-0.95(m,2H). LCMS:[M+H] + = 920.4.

[0188] Example 3: Synthesis of 5-(4-(4-((4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyridino[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)piperazine-1-yl)-2-(1-methyl-2,6-dioxopiperidine-3-yl)isoindoline-1,3-dione (Me-dBPTF-2)

[0189] [ka]

[0190] In a solution of 4-(4-(2-(1-methyl-2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-5-yl)piperazine-1-yl)butanal (89 mg, 0.21 mmol) in DCE (1 mL), 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyridino[1,2-a]pyrimidine-4-one (109.8 mg, 0.21 mmol) and DIEA (41.3 mg) were added, and the reaction mixture was stirred at room temperature for 30 minutes. Then, STAB (133.5 mg, 0.63 mmol) was added, and the reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was diluted with water (50 mL) and extracted with DCM (20 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, and concentrated to obtain a residue, which was purified by preparative HPLC to obtain the title compound as a yellow solid (28 mg, yield 14%). 1H NMR(400MHz,DMSO)δ9.00(s,1H),8.47(d,J=6.3Hz,2H),8.14-7.94(m,3H),7.71-7.51(m,3H),7.40(d,J=9. 1Hz,1H),7.28(s,1H),7.21(d,J=8.8Hz,1H),5.13(dd,J=13.1,5.4Hz,1H),3.79-3.72(m,1H),3.42-3.35(m ,4H),3.00(s,3H),2.99-2.92(m,4H),2.81-2.64(m,2H),2.57-2.53(m,1H),2.47-2.37(m,8H),2.34-2.23( LCMS:[M+H] + =934.3.

[0191] Example 4: Synthesis of 5-(4-(6-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyridino[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)hexyl)piperazine-1-yl)-2-(2,6-dioxopiperidine-3-yl)isoindoline-1,3-dione (dBPTF-9, 1B-2)

[0192] [ka]

[0193] 6-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-5-yl)piperazine-1-yl)hexanal In 30 mL of ACN, a solution of 2-(2,6-dioxopiperidine-3-yl)-5-(4-(6-hydroxyhexyl)piperazine-1-yl)isoindoline-1,3-dione (100 mg, 0.226 mmol) was added to IBX (127 mg, 0.454 mmol), and the reaction mixture was stirred at 80°C for 2 hours. The reaction mixture was diluted with water (30 mL) and extracted with siRNA (30 mL x 3). The combined organic layers were concentrated under reduced pressure to obtain a residue, which was purified by silica gel column elution with MeOH:DCM (1:20) to obtain the title compound as a yellow solid (67 mg, 67%). LCMS: [M+H] + = 441.2.

[0194] 5-(4-(6-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)hexyl)piperazine-1-yl)-2-(2,6-dioxopiperidine-3-yl)isoindorin-1,3-dione In a solution of 6-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-5-yl)piperazine-1-yl)hexanal (60 mg, 0.136 mmol) and 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (85 mg, 0.136 mmol) in DCE (5 mL), HOAc (19.1 mg, 0.318 mmol) was added, and the reaction mixture was stirred for 30 minutes. Then, STAB (101 mg, 0.476 mmol) was added, and the mixture was stirred for 4 hours. The reaction mixture was diluted with water (30 mL) and extracted with DCM (30 mL x 3). The combined organic layers were concentrated under reduced pressure to obtain a residue, which was purified by preparative HPLC (5-95% acetonitrile containing 0.1% NH3·H2O in water) to obtain the title compound as a yellow solid (8 mg, 14%). 1H NMR(400MHz,DMSO)δ11.07(s,1H),9.01(s,1H),8.46(s,2H),8.14-7.96(m,3H),7.71-7.50(m,3 H),7.41(d,J=9.2Hz,1H),7.29(s,1H),7.21(d,J=8.5Hz,1H),5.12-5.00(m,1H),3.84-3.72(m,2 H),3.05-2.79(m,7H),2.69-2.56(m,5H),2.47-2.40(m,6H),2.26(t,J=6.8Hz,4H),2.18(s,3H) ,2.05-1.97(m,1H),1.46-1.31(m,4H),1.30-1.20(m,4H),1.13-1.06(m,2H),1.03-0.94(m,2H). LCMS:[M+H] + =948.5.

[0195] Example 5: Synthesis of 5-(4-(2-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)ethyl)piperazine-1-yl)-2-(2,6-dioxopiperidine-3-yl)isoindoline-1,3-dione (dBPTF-10, 1B-3)

[0196] [ka]

[0197] A solution of 2-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-5-yl)piperazine-1-yl)acetaldehyde (18 mg, 0.048 mmol), 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (50 mg, 0.096 mmol), and STAB (20.5 mg, 0.096 mmol) in THF (20 mL) was stirred at room temperature for 20 minutes. Furthermore, 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (18 mg, 0.048 mmol) and STAB (20.5 mg, 0.096 mmol) were added. The reaction mixture was diluted with H2O (30 mL) and extracted by DCM (30 mL x 3). The combined organic layers were concentrated under reduced pressure to obtain a residue, which was then subjected to preparative HPLC (0.1% NH3 in water). · The title compound was purified with acetonitrile containing H2O, 5% to 95%, to obtain it as a yellow solid (20 mg, 18%). 1 H NMR(400MHz,DMSO)δ11.06(s,1H),9.01(s,1H),8.46(s,2H),8.14-7.93(m,3H),7.69-7 .48(m,3H),7.41(d,J=9.0Hz,1H),7.30(s,1H),7.22(d,J=9.2Hz,1H),5.05(dd,J=12.7, 5.4Hz,1H),3.77(s,1H),3.52-3.35(m,7H),3.09-2.79(m,6H),2.61-2.55(m,5H),2.48 -2.41(m,5H),2.19(s,3H),2.06-1.92(m,1H),1.16-1.05(m,2H),1.00(d,J=5.7Hz,2H). LCMS:[M+H] + = 892.3.

[0198] Example 6: Synthesis of 4-(4-(8-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)octyl)piperazine-1-yl)-2-(2,6-dioxopiperidine-3-yl)isoindoline-1,3-dione (dBPTF-13, NVS-BPTF-A-2)

[0199] [ka]

[0200] tert-butyl4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-4-yl)piperazine-1-carboxylate To a solution of 2-(2,6-dioxopiperidine-3-yl)-4-fluoroisoindoline-1,3-dione (10 g, 36.2 mmol) in DMSO (50 mL), tert-butylpiperazine-1-carboxylate (10.11 g, 54 mmol) and DIEA (14 g, 108.6 mmol) were added. The mixture was heated to 130 °C and stirred for 16 hours. The solution was diluted with water (50 mL) and extracted with dimethyl phosphate (80 mL x 3). The combined organic layers were washed with brine (40 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column elution with DCM:MeOH (0-10%) to obtain the title compound as a green solid (13 g, 81%). 1 H NMR(400MHz,DMSO)δ11.10(s,1H),7.73(dd,J=8.3,7.2Hz,1H),7.38(dd,J=15.2,7.7Hz,2H),5.11(dd,J=12.8,5.4Hz,1H ),3.52(s,4H),3.26(d,J=4.3Hz,4H),2.95-2.81(m,1H),2.66-2.52(m,2H),2.08-1.99(m,1H),1.43(s,9H).LCMS:[M+H] + = 443.0.

[0201] 2-(2,6-dioxopiperidine-3-yl)-4-(piperazine-1-yl)isoindorin-1,3-dione To a solution of tert-butyl 4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-4-yl)piperazine-1-carboxylate (5 g, 11.31 mmol) in dioxane (20 mL), dioxane / HCl (4 mL) was added. The mixture was stirred at room temperature for 16 hours, and then concentrated under reduced pressure to obtain the title compound as a yellow solid (5.1 g, crude), which was used in the next step without further purification. LCMS:[M+H] + = 343.1.

[0202] 2-(2,6-dioxopiperidine-3-yl)-4-(4-(8-hydroxyoctyl)piperazine-1-yl)isoindorin-1,3-dione In 40 mL of ACN, a solution of 2-(2,6-dioxopiperidine-3-yl)-4-(piperazine-1-yl)isoindorin-1,3-dione (4.5 g, 13.15 mmol) was prepared, to which 8-bromooctan-1-ol (3.026 g, 14.47 mmol), K2CO3 (5.4 g, 39 mmol), and KI (1.941 g, 13.15 mmol) were added. The mixture was heated to 75°C, stirred for 16 hours, diluted with water (40 mL), extracted with DCM:MeOH = 10:1 (50 mL x 3), the organic layers were combined, washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column elution with DCM:MeOH (0-5%) to obtain the title compound as a green solid (2.7g, 43%). 1H NMR(400MHz,DMSO)δ11.09(s,1H),7.70(dd,J=8.3,7.3Hz,1H),7.35(t,J=8.0H z,2H),5.09(dd,J=12.7,5.4Hz,1H),4.37(t,J=5.2Hz,1H),3.36(d,J=6.5Hz,3H ),3.30(s,4H),2.86(dd,J=8.5,5.4Hz,1H),2.64-2.53(m,5H),2.34(s,2H),2.0 3(dd,J=9.0,3.6Hz,1H),1.50-1.38(m,4H),1.26(d,J=10.6Hz,9H).LCMS:[M+H] + = 471.1.

[0203] 8-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-4-yl)piperazine-1-yl)octanal A solution of 2-(2,6-dioxopiperidine-3-yl)-4-(4-(8-hydroxyoctyl)piperazine-1-yl)isoindorin-1,3-dione (2.5 g, 5.32 mmol) and PCC (2.3 g, 10.6 mmol) in DCM (20 mL) was stirred at room temperature for 4 hours. The mixture was diluted with water (50 mL) and extracted with DCM:MeOH = 10:1 (50 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column elution with DCM:MeOH (0-5%) to obtain the title compound as a yellow solid (1.03 g, 41%). LCMS:[M+H] + = 469.2.

[0204] 4-(4-(8-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyridino[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)octyl)piperazine-1-yl)-2-(2,6-dioxopiperidine-3-yl)isoindoline-1,3-dione (dBPTF-13, NVS-BPTF-A-2) Sodium triacetoxyborohydride (1.7 g, 7.94 mmol) was added to a solution of 8-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-4-yl)piperazine-1-yl)octanal (930 mg, 1.98 mmol) and 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (1.039 mg, 1.98 mmol) in DCE (20 mL). The mixture was stirred at room temperature for 16 hours, then diluted with water (30 mL), extracted with DCM (40 mL x 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was subjected to preparative HPLC (0.1% NH3 in water). · The title compound was purified with acetonitrile containing H2O (5% to 95%) to obtain a yellow solid (324.3 mg, 17%). 1 H NMR(400MHz,DMSO)δ11.08(s,1H),9.01(d,J=1.9Hz,1H),8.49-8.45(m,2H),8.08(dd,J=9.2,2.1Hz,1H),8.04-7.97(m, 2H),7.71-7.65(m,1H),7.64-7.52(m,2H),7.41(d,J=9.1Hz,1H),7.32(dd,J=13.9,7.7Hz,2H),5.08(dd,J=12.9,5.4Hz, 1H),3.81-3.73(m,1H),3.32-3.21(m,7H),2.99-2.82(m,5H),2.63-2.52(m,3H),2.46-2.38(m,4H),2.31-2.22(m,4H), 2.18(s,3H),2.06-1.95(m,1H),1.47-1.31(m,4H),1.27-1.18(m,8H),1.12-1.07(m,2H),1.03-0.96(m,2H).LCMS:[M+H] + =976.1.

[0205] Example 7: Synthesis of 4-(4-(8-(4-((4-((7-(1-cyclopropyl-1H-pyrazol-4-yl)-3-methyl-4-oxo-4H-pyridino[1,2-a]pyrimidin-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazin-1-yl)octyl)piperazin-1-yl)-2-(1-methyl-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (Me-dBPTF-13)

[0206]

Chemical Formula

[0207] To a solution of 8-(4-(2-(1-methyl-2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperazin-1-yl)octanal (20 mg, 0.043 mmol) and 7-(1-cyclopropyl-1H-pyrazol-4-yl)-2-((2-fluoro-4-(piperazin-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidin-4-one (23 mg, 0.043 mmol) in DCE (10 mL) was added STAB (36 mg, 0.17 mmol), and the reaction mixture was stirred at room temperature for 4 hours. Water (20 mL) was added, the mixture was extracted with DCM (10 mL × 3), dried over Na2SO4, and concentrated to obtain a residue, which was purified by preparative HPLC (0.1% NH3 in water · containing H2O, acetonitrile 5%~95%) to give the title compound as a yellow solid (8 mg, yield 19%). 1H NMR(400MHz,DMSO)δ9.01(s,1H),8.46(s,2H),8.08(dd,J=9.3,1.9Hz,1H),8.04-7.97(m,2H),7.68(t,J=7.7Hz,1H),7.57(dd,J=22.4, 9.9Hz,2H),7.41(d,J=9.1Hz,1H),7.32(dd,J=12.0,7.8Hz,2H),5.15(dd,J=13.1,5.3Hz,1H),3.76(m,J=11.1,3.9Hz,1H),3.29-3.23(m ,4H),3.01(s,3H),2.98-2.92(m,4H),2.79-2.71(m,J=17.0Hz,1H),2.59-2.52(m,4H),2.46-2.42(m,6H),2.30-2.22(m,J=15.1,8.4Hz ,4H),2.18(s,3H),2.06-2.00(m,1H),1.43-1.33(m,4H),1.26-1.19(m,8H),1.12-1.06(m,2H),1.02-0.96(m,J=7.2Hz,2H).LCMS:[M+H] + = 990.8.

[0208] Example 8: Synthesis of 4-(4-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)octyl)piperazine-1-yl)-2-(2,6-dioxopiperidine-3-yl)isoindoline-1,3-dione (dBPTF-14)

[0209] [ka]

[0210] In a solution of 4-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-4-yl)piperazine-1-yl)butanal (100 mg, 0.24 mmol) and 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (139 mg, 0.26 mmol) in DCE (10 mL), STAB (36 mg, 0.17 mmol) was added, and the reaction mixture was stirred at room temperature for 4 hours. Water (20 mL) was added, and the mixture was extracted with DCM (10 mL x 3), dried over Na2SO4, and concentrated to obtain a residue, which was then subjected to preparative HPLC (0.1% NH3 in water). · The title compound was purified with acetonitrile containing H2O (5% to 95%) to obtain a yellow solid (22 mg, 10% yield). 1 H NMR(400MHz,DMSO)δ8.99(s,1H),8.44(s,1H),8.02(d,J=9.1Hz,1H),7.99-7.89(m,2H),7.72-7.64( m,1H),7.57-7.44(m,2H),7.39-7.28(m,3H),7.28-7.20(m,4H),5.05(dd,J=13.1,5.9Hz,1H),3.80-3 .72(m,1H),3.49(d,J=10.6Hz,4H),3.28-3.25(m,4H),3.02-2.88(m,4H),2.85-2.74(m,1H),2.62-2 .55(m,2H),2.47-2.38(m,7H),2.16(s,3H),2.02-1.96(m,2H),1.12-1.07(m,2H),1.03-0.96(m,2H). LCMS:[M+H] + = 920.4.

[0211] Example 9: Synthesis of 4-(4-((4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)methyl)benzyl)piperazine-1-yl)-2-(2,6-dioxopiperidine-3-yl)isoindoline-1,3-dione (dBPTF-15)

[0212] [ka]

[0213] In a solution of 4-((4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-4-yl)piperazine-1-yl)methyl)benzaldehyde (26 mg, 0.057 mmol) and 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (30 mg, 0.057 mmol) in MeOH (9 mL), Pic-BH3 (6 mg, 0.057 mmol) was added, and the reaction mixture was stirred at room temperature for 2 hours. Water (20 mL) was added, and the mixture was extracted with DCM (10 mL x 3), dried over Na2SO4, and concentrated to obtain a residue, which was then subjected to preparative HPLC (0.1% NH3 in water). · The title compound was purified with acetonitrile containing H2O (5% to 95%) to obtain a yellow solid (6 mg, 10% yield). 11H NMR (400 MHz, DMSO) δ 11.08 (s, 1H), 9.01 (d, J=1.7 Hz, 1H), 8.51-8.42 (m, J=5.4 Hz, 2H), 8.12-8.11 (m, J=11.2 Hz, 1H), 8.04-7.95 (m, 2H), 7.70-7.52 (m, 3H), 7.41 (d, J=9.0 Hz, 1H), 7.41 (d, J=9.0 Hz, 1H), 7.33 (d, J=7.4 Hz, 1H), 5.07 (dd, J=12.7, 5.3 Hz, 1H), 3.83-3.72 (m, 1H), 3.32-3.29 (m, 4H), 3.26-3.21 (m, 4H), 3.00-2.90 (m, J=7.1, 4.7 Hz, 4H), 2.88-2.80 (m, 1H), 2.62-2.53 (m, 3H), 2.46-2.38 (m, J=8.0, 6.7 Hz, 4H), 2.32-2.26 (m, J=5.0, 3.9 Hz, 3H), 2.18 (s, 3H), 2.04-1.96 (m, 1H), 1.50-1.35 (m, 4H), 1.13-1.05 (m, 2H), 1.05-0.96 (m, 2H). LCMS: [M+H] + =968.2.

[0214] Example 10: Synthesis of 3-(6-(7-(4-(4-((4-((7-(1-cyclopropyl-1H-pyrazol-4-yl)-3-methyl-4-oxo-4H-pyridino[1,2-a]pyrimidin-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazin-1-yl)butyl)-2,7-diazaspiro[3.5]nonan-2-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (dBPTF-16)

[0215]

Chemical Formula

[0216] tert-butyl 2-(2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)-2,7-diazaspiro[3.5]nonane-7-carboxylate To a solution of 3-(6-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (2.5 g, 7.7 mmol) in dioxane (50 mL), tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate (1.7 g, 7.7 mmol), PEPPSI-Pd (749 mg, 0.77 mmol), and Cs2CO3 (7.6 g, 23.2 mmol) were added. The reaction mixture was stirred overnight at 100°C. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (120 mL x 3). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using elution with 5% MeOH in DCM to obtain the title compound as a white solid (1 g, 28%). LCMS: [M+H] + = 469.2.

[0217] 3-(1-oxo-6-(2,7-diazaspiro[3.5]nonane-2-yl)isoindoline-2-yl)piperidine-2,6-dione In 10 mL of DCM, a solution of tert-butyl 2-2-(2,6-dioxopiperidine-3-yl)-3-oxoisoindorin-5-yl)-2,7-diazaspiro[3.5]nonane-7-carboxylate was added, to which 5 mL of TFA was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a white solid (200 mg, crude), which was used without further purification. LCMS: [M+H] + =369.1.

[0218] 3-(6-(7-(3-(1,3-dioxolan-2-yl)propyl)-2,7-diazaspiro[3.5]nonan-2-yl)-1-oxoisoindorin-2-yl)piperidine-2,6-dione In a solution of 3-(1-oxo-6-(2,7-diazaspiro[3.5]nonan-2-yl)isoindorin-2-yl)piperidine-2,6-dione (200 mg, 0.54 mmol) in ACN (30 mL), 2-(3-bromopropyl)-1,3-dioxolane (117 mg, 0.60 mmol), K2CO3 (298 mg, 2.16 mmol), and KI (99 mg, 0.60 mmol) were added, and the reaction mixture was stirred overnight at 80°C. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using elution with 5%-10% MeOH in DCM to obtain the title compound as a white solid (130 mg, 50%). LCMS:[M+H] + = 483.2.

[0219] 4-(2-(2-(2,6-dioxopiperidine-3-yl)-3-oxoisoindorin-5-yl)-2,7-diazaspiro[3.5]nonane-7-yl)butanal In a solution of 3-(6-(7-(3-(1,3-dioxolan-2-yl)propyl)-2,7-diazaspiro[3.5]nonan-2-yl)-1-oxoisoindorin-2-yl)piperidine-2,6-dione (190 mg, 0.39 mmol) in DCM (10 mL), TFA (5 mL) was added, and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a white solid (190 mg, crude), which was used without further purification. LCMS:[M+H] + = 439.2.

[0220] 3-(6-(7-(4-(4-((4-((7-(1-Cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)-2,7-diazaspiro[3.5]nonane-2-yl)-1-oxoisoindorin-2-yl)piperidine-2,6-dione In a solution of 4-(2-(2-(2,6-dioxopiperidine-3-yl)-3-oxoisoindorin-5-yl)-2,7-diazaspiro[3.5]nonan-7-yl)butanal (84 mg, 0.16 mmol) and 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (70 mg, 0.16 mmol) in DCE (10 mL), STAB (144 mg, 0.6317 mmol) was added, and the reaction mixture was stirred at room temperature for 4 hours. Water (20 mL) was added, and the mixture was extracted with DCM (10 mL x 3), dried over Na2SO4, and concentrated to obtain a residue, which was then subjected to preparative HPLC (0.1% NH3 in water). · The title compound was purified with acetonitrile containing H2O (5% to 95%) to obtain a yellow solid (17 mg, yield 11%). 1 H NMR(400MHz,DMSO)δ10.97(s,1H),9.01(s,1H),8.47(s,2H),8.09(d,J=9.2Hz,1H),8.07-8.00(m,2H),7.60(dd,J=23.4,10.1Hz,2 H),7.41(d,J=9.2Hz,1H),7.35(d,J=8.8Hz,1H),6.64(s,2H),5.07(dd,J=13.2,5.2Hz,1H),4.34-4.13(m,2H),3.82-3.72(m,1H), 3.02-2.93(m,4H),3.02-2.93(m,4H),2.91-2.85(m,1H),2.64-2.56(m,2H),2.48-2.42(m,4H),2.39-2.33(m,2H),2.32-2.23(m,4) LCMS:[M+H] + =946.3.

[0221] Example 11: Synthesis of 3-((4-(4-(6-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)hexyl)piperazine-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (dBPTF-17)

[0222] [ka]

[0223] tert-butyl 4-(2-fluoro-4-nitrophenyl)piperazine-1-carboxylate In 50 mL of DMF, a solution of 1,2-difluoro-4-nitrobenzene (40 g, 251 mmol) was added to tert-butylpiperazine-1-carboxylate (46.7 g, 251 mmol) and K2CO3 (103.9 g, 753 mmol). The reaction mixture was stirred at 130°C for 2 hours. The reaction mixture was diluted with water (150 mL) and extracted with ethyl acetate (60 mL x 3). The combined organic layers were washed with brine (90 mL), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using elution with 20% ethyl acetate in petroleum ether to obtain the title compound as a yellow solid (80 g, 98%). LCMS: [M+H] + = 270.1.

[0224] tert-butyl 4-(4-amino-2-fluorophenyl)piperazine-1-carboxylate In 20 mL of MeOH, a solution of tert-butyl 4-(2-fluoro-4-nitrophenyl)piperazine-1-carboxylate (10 g, 30.77 mmol) was added, to which Pd / C (1 g) was added. The reaction mixture was stirred overnight at room temperature. The reaction mixture was filtered through Celite® and washed with ethyl acetate. The filtrate was concentrated to obtain the title compound as a yellow solid (8.5 g, crude). LCMS: [M+H] + = 296.2.

[0225] tert-butyl 4-(4-((2,6-dioxopiperidine-3-yl)amino)-2-fluorophenyl)piperazine-1-carboxylate To a solution of tert-butyl 4-(4-amino-2-fluorophenyl)piperazine-1-carboxylate (8.5 g, 28.81 mmol) in DMSO (20 mL), DIEA (19.1 mL) and 3-bromopiperidine-2,6-dione (6.74 g, 34.57 mmol) were added, and the reaction mixture was stirred at 130 °C for 12 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using elution with 5% MeOH in DCM to obtain the title compound as a green solid (3.5 g, 30%). LCMS: [M+H] + = 407.2.

[0226] 3-((3-fluoro-4-(piperazine-1-yl)phenyl)amino)piperidine-2,6-dione To a solution of tert-butyl 4-(4-((2,6-dioxopiperidine-3-yl)amino)-2-fluorophenyl)piperazine-1-carboxylate (700 mg, 1.72 mmol) in dioxane (5 mL), HCl / dioxane (5 mL) was added at 0°C, and the reaction mixture was stirred for 4 hours. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a green solid (525 mg, crude), which was used without further purification. LCMS:[M+H] + = 307.2.

[0227] 6-(4-(4-((2,6-dioxopiperidine-3-yl)amino)-2-fluorophenyl)piperazine-1-yl)hexanoic acid In 10 mL of ACN, a solution of 3-((3-fluoro-4-(piperazin-1-yl)phenyl)amino)piperidine-2,6-dione (525 mg, 1.72 mmol) was mixed with DIEA (885 mg, 6.86 mmol), 6-bromohexanoic acid (1.00 g, 5.16 mmol), and KI (314 mg, 1.89 mmol). The reaction mixture was stirred overnight at 80°C. The reaction mixture was concentrated to obtain a residue, which was purified by silica gel column elution with 20% MeOH in DCM to obtain the title compound as a green solid (402 mg, 56%). LCMS: [M+H] + = 421.1.

[0228] S-Octyl 6-(4-(4-((2,6-Dioxopiperidine-3-yl)amino)-2-fluorophenyl)piperazine-1-yl)Hexanethionate In a solution of 6-(4-(4-((2,6-dioxopiperidine-3-yl)amino)-2-fluorophenyl)piperazine-1-yl)hexanoic acid (402 mg, 0.96 mmol) in DCM (3 mL), octan-1-thiol (116.8 mg, 0.8 mmol), EDCI (183.4 mg, 0.96 mmol), and DMAP (19.5 mg, 0.16 mmol) were added at 0°C and the mixture was stirred overnight. The reaction mixture was concentrated to obtain a residue, which was purified by silica gel column elution with 20% MeOH in DCM to obtain the title compound as a green solid (162 mg, 34%). LCMS:[M+H] + = 549.4.

[0229] 6-(4-(4-((2,6-dioxopiperidine-3-yl)amino)-2-fluorophenyl)piperazine-1-yl)hexanal In 5 mL of DCM, a solution of S-octyl 6-(4-(4-((2,6-dioxopiperidine-3-yl)amino)-2-fluorophenyl)piperazin-1-yl)hexanethionate (90 mg, 0.16 mmol) was added, to which Pd(OAc)2 (3.6 mg) and Et3SiH (37.21 mg, 0.32 mmol) were added, and the mixture was stirred overnight at room temperature. The reaction mixture was filtered through a Celite® pad and washed with DCM. The filtrate was purified by preparative TLC to obtain the title compound as a green solid (32 mg, 50%). LCMS:[M+H] + = 405.2.

[0230] 3-((4-(4-(6-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)hexyl)piperazine-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione In a solution of 6-(4-(4-((2,6-dioxopiperidine-3-yl)amino)-2-fluorophenyl)piperazin-1-yl)hexanal (32 mg, 0.079 mmol) in DCE (5 mL), 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazin-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (41.4 mg, 0.079 mmol) and DIEA (15 mg) were added, and the reaction mixture was stirred at room temperature for 30 minutes. STAB (200 mg, 0.94 mmol) was added, and the reaction mixture was then stirred at room temperature for 30 minutes. The reaction mixture was diluted with water (50 mL) and extracted with DCM (20 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by preparative HPLC to obtain the title compound as a white solid (15 mg, 21%). 1H NMR(400MHz,DMSO)δ10.77(s,1H),9.01(s,1H),8.51-8.42(m,2H),8.08(d,J=9.0Hz,1H),8.05-7.96(m,2H),7.61(d,J=10.1Hz,1H),7. 55(d,J=8.7Hz,1H),7.41(d,J=9.2Hz,1H),6.79(t,J=9.2Hz,1H),6.49(d,J=15.1Hz,1H),6.40(d,J=9.2Hz,1H),5.78(d,J=7.9Hz,1H),4 .29-4.18(m,1H),3.80-3.72(m,1H),3.02-2.89(m,4H),2.86-2.76(m,4H),2.76-2.56(m,3H),2.46-2.37(m,7H),2.29-2.20(m,4H),2.1 9(s,3H),2.12-2.02(m,1H),1.88-1.80(m,1H),1.45-1.33(m,4H),1.28-1.18(m,4H),1.13-1.05(m,2H),1.05-0.96(m,2H).LCMS:[M+H] + = 912.7.

[0231] Example 12: Synthesis of 3-((4-(4-(1-(4-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)-3,3-difluoropiperazine-4-yl)piperazine-1-yl)-3-fluorophenyl)amino)piperidine-2,6-dione (dBPTF-18, A-3)

[0232] [ka]

[0233] In 10 mL of DCE, a solution of 4-(4-(4-(4-((2,6-dioxopiperidine-3-yl)amino)-2-fluorophenyl)piperazine-1-yl)-3,3-difluoropiperidine-1-yl)butanal (42 mg, 0.08 mmol) and 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (39.7 mg, 0.08 mmol) was added, and STAB (57 mg, 0.27 mmol) was added. The reaction mixture was stirred overnight at room temperature. 50 mL of water was added, and the mixture was extracted with DCM (30 mL x 3), dried over Na2SO4, and concentrated to obtain a residue. This residue was then subjected to preparative HPLC (0.1% NH3 in water). · The title compound was purified with acetonitrile containing H2O (5% to 95%) to obtain a white solid (11 mg, yield 13%). 1 H NMR(400MHz,MeOH)δ9.06(s,1H),8.47(t,J=8.1Hz,1H),8.23(s,1H),8.03(d,J=9.1Hz,1H),7.93(s,1H),7.70-7.62(m,2H) ),7.46(d,J=9.2Hz,1H),6.83(t,J=8.9Hz,1H),6.50-6.37(m,2H),4.23-4.15(m,1H),3.76-3.68(m,1H),3.39-3.34(m,6H) ),3.26-3.20(m,2H),3.17-3.10(m,3H),3.09-2.96(m,8H),2.85-2.63(m,5H),2.62-2.36(m,2H),2.31-2.19(m,4H),2.12 -2.00(m,1H),1.96-1.86(m,2H),1.84-1.74(m,2H),1.73-1.63(m,2H),1.38-1.24(m,1H),1.19-1.05(m,4H).LCMS:[M+H] + = 1003.5.

[0234] Example 13: Synthesis of 3-((4-(4-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)piperazine-1-yl)3-fluorophenyl)amino)piperidine-2,6-dione (dBPTF-19, A-4)

[0235] [ka]

[0236] A 10 mL solution of DCE (10 mL) containing 4-(4-(4-((2,6-dioxopiperidine-3-yl)amino)-2-fluorophenyl)piperazine-1-yl)butanal (50 mg, 0.13 mmol), 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (35 mg, 0.07 mmol), and DIEA (0.3 mL) was stirred at room temperature for 30 minutes. STAB (57 mg, 0.27 mmol) was added, and the reaction mixture was stirred at room temperature overnight. Water (50 mL) was added, extracted with DCM (30 mL x 3), dried with Na2SO4, concentrated to obtain a residue, which was then subjected to preparative HPLC (0.1% NH3 in water). · The title compound was purified with acetonitrile containing H2O (5% to 95%) to obtain a white solid (11 mg, 9% yield). 1H NMR(400MHz,MeOH)δ9.08(d,J=1.8Hz,1H),8.25-8.18(m,2H),8.05-8.00(m,1H),7.93(s,1H),7 .65-7.55(m,2H),7.48-7.41(m,1H),6.79(t,J=8.9Hz,1H),6.45-6.35(m,2H),4.25-4.15(m,1H) ),3.77-3.69(m,1H),3.38-3.31(m,4H),3.24-3.00(m,10H),2.84-2.65(m,6H),2.54(t,J=6.6H z,2H),2.34-2.17(m,4H),1.97-1.74(m,3H),1.71-1.58(m,2H),1.19-1.06(m,4H).LCMS:[M+H] + = 884.4.

[0237] Example 14: Synthesis of N-(4-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)-3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoamine (dBPTF-20)

[0238] [ka]

[0239] tert-butyl(4-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)carbamate In 3 mL of NMP, a solution of 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (100 mg, 0.19 mmol) was added, to which tert-butyl 4-bromobutyl)carbamate (289 mg, 1.15 mmol), KI (63 mg, 0.38 mmol), and K2CO3 (262 mg, 1.9 mmol) were added. The reaction mixture was stirred in a microwave at 150°C for 2 hours. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using elution with 20% MeOH in DCM ether to obtain the title compound as a white solid (60 mg, 45%). LCMS:[M+H] + = 695.4.

[0240] 2-((4-((4-(4-aminobutyl)piperazine-1-yl)sulfonyl)-2-fluorophenyl)amino)-7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one In a solution of tert-butyl(4-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)carbamate (50 mg, 0.072 mmol) in dichloromethane (2 mL), HCl / dioxane (2 mL, 4 M) was added, and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a brown solid (50 mg, crude), which was used without further purification. LCMS:[M+H] + = 595.2.

[0241] N-(4-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)-3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzamide In 5 mL of DCM, a solution of 2-((4-((4-(4-aminobutyl)piperazin-1-yl)sulfonyl)-2-fluorophenyl)amino)-7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (35 mg, 0.059 mmol), 3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoic acid (25 mg, 0.059 mmol), and DIPEA (38 mg, 0.295 mmol) was mixed with HATU (30 mg, 0.078 mmol). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with water and extracted with DCM. The combined organic layer was concentrated to obtain a residue, which was purified by preparative HPLC to obtain the title compound as a white solid (10 mg, 14%). 1 H NMR(400MHz,DMSO)δ10.34(s,1H),9.02(d,J=2.0Hz,1H),8.47(d,J=3.2Hz,2H),8.30(d,J=5.4Hz,1H),8.08(dd,J=9.2, 2.1Hz,1H),8.03-7.96(m,2H),7.82(dd,J=8.6,2.2Hz,1H),7.74(d,J=2.2Hz,1H),7.64-7.50(m,2H),7.41(d,J=9.3Hz, 1H),7.14(d,J=8.8Hz,1H),3.83(s,3H),3.80-3.73(m,1H),3.60-3.54(m,2H),3.23-3.18(m,2H),2.95(s,4H),2.69-2. 65(m,2H),2.43(s,4H),2.33-2.27(m,2H),2.18(s,3H),1.51-1.38(m,4H),1.14-1.06(m,2H),1.00(m,2H).LCMS:[M+H] + = 841.4.

[0242] Example 15: Synthesis of N-(6-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)-3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoamine (dBPTF-21)

[0243] [ka]

[0244] tert-butyl(6-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)hexyl)carbamate In 5 mL of DCE, a solution of 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (200 mg, 0.38 mmol) was added, to which DIEA (74 mg, 0.57 mmol) was added. The reaction mixture was stirred at room temperature for 30 minutes. In 5 mL of DCE, a solution of tert-butyl(6-oxohexyl)carbamate (83 mg, 0.38 mmol) and STAB (242 mg, 1.14 mmol) was added, and the reaction mixture was stirred overnight. The reaction mixture was diluted with water and extracted with DCM. The combined organic layers were dried over Na2SO4 and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using elution with 10% MeOH in DCM to obtain the title compound as a yellow solid (120 mg, 44%). LCMS:[M+H] + = 723.4.

[0245] 2-((4-((4-(6-aminohexyl)piperazine-1-yl)sulfonyl)-2-fluorophenyl)amino)-7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one In a solution of tert-butyl(6-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazin-1-yl)hexyl)carbamate (120 mg, 0.17 mmol) in DCM (5 mL), HCl / dioxane (5 mL, 4 M) was added, and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a yellow solid (100 mg, crude), which was used without further purification. LCMS:[M+H] + = 623.8.

[0246] N-(6-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)hexyl)-3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzamide In 5 mL of DCM, a solution of 2-((4-((4-(6-aminohexyl)piperazin-1-yl)sulfonyl)-2-fluorophenyl)amino)-7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4H-pyridino[1,2-a]pyrimidine-4-one (100 mg, 0.16 mmol) was mixed with DIEA (207 mg, 1.61 mmol) and perfluorophenyl 3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoate (138 mg, 0.32 mmol). The reaction mixture was stirred at room temperature for 20 minutes. The reaction mixture was diluted with water and extracted with DCM. The combined organic layer was concentrated to obtain a residue, which was purified by preparative HPLC to obtain the title compound as a white solid (28 mg, 18%). 1H NMR(400MHz,DMSO)δ10.34(s,1H),9.01(d,J=1.9Hz,1H),8.54-8.40(m,2H),8.31(t,J=5.6Hz,1H),8.14-8.05(m,1H),8.04-7.98(m,2 H),7.82(dd,J=8.7,2.2Hz,1H),7.75(d,J=2.2Hz,1H),7.61(d,J=10.4Hz,1H),7.58-7.51(m,1H),7.41(d,J=9.2Hz,1H),7.15(d,J=8. 8Hz,1H),3.84(s,3H),3.80-3.73(m,1H),3.58(t,J=6.5Hz,2H),3.24-3.16(m,2H),3.07-2.81(m,4H),2.72-2.64(m,2H),2.49-2.35( m,4H),2.35-2.21(m,2H),2.18(s,3H),1.53-1.43(m,2H),1.42-1.32(m,2H),1.31-1.21(m,4H),1.14-1.05(m,2H),1.05-0.95(m,2H). LCMS:[M+H] + = 869.6.

[0247] Example 16: Synthesis of 6-(4-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)-2-(2,6-dioxopiperidine-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione (dBPTF-22)

[0248] [ka]

[0249] 6-(3-(1,3-dioxolan-2-yl)propyl)-2-(2,6-dioxopiperidine-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione In 8 mL of ACN, a solution of 2-(2,6-dioxopiperidine-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione (66 mg, 0.22 mmol) was mixed with DIEA (114 mg, 0.88 mmol), 2-(3-bromopropyl)-1,3-dioxolane (129 mg, 0.66 mmol), and KI (40 mg, 0.24 mmol). The reaction mixture was stirred in a microwave at 80°C for 1 hour. The reaction mixture was concentrated to obtain a residue, which was purified by flash silica gel chromatography (10% MeOH in DCM) to obtain the title compound as a yellow solid (90 mg, 99%). LCMS: [M+H] + = 414.1.

[0250] 4-(6-(2,6-dioxopiperidine-3-yl)-5,7-dioxo-3,5,6,7-tetrahydropyrrolo[3,4-f]isoindole-2(1H)-yl)butanal In a solution of 6-(3-(1,3-dioxolan-2-yl)propyl)-2-(2,6-dioxopiperidine-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione (90 mg, 0.22 mmol) in DCM (4 mL), TFA (3 mL) was added, and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a yellow solid (70 mg, crude), which was used without further purification. LCMS: [M+H] + =370.1.

[0251] 6-(4-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)-2-(2,6-dioxopiperidine-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione In a solution of 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (76 mg, 0.15 mmol) in DCE (5 mL), DIEA (285 mg, 0.22 mmol) was added, and the reaction mixture was stirred at room temperature for 30 minutes. 4-(6-(2,6-dioxopiperidine-3-yl)-5,7-dioxo-3,5,6,7-tetrahydropyrrolo[3,4-f]isoindole-2(1H)-yl)butanal (70 mg, 0.19 mmol) and STAB (93 mg, 0.44 mmol) were added, and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with water and extracted with DCM. The combined organic layers were concentrated to obtain a residue, which was purified by preparative HPLC to obtain the title compound as a white solid (11 mg, 8%). 1 H NMR(400MHz,DMSO)δ11.10(s,1H),9.00(d,J=1.9Hz,1H),8.53-8.42(m,2H),8.10-8.00(m,2H),7.99(s,1H),7.73(s,2H) ),7.61(dd,J=10.2,1.9Hz,1H),7.56(dd,J=8.5,1.8Hz,1H),7.39(d,J=9.2Hz,1H),5.12(dd,J=12.9,5.4Hz,1H),3.90(s ,4H),3.80-3.73(m,1H),3.03-2.92(m,4H),2.92-2.83(m,1H),2.68-2.60(m,3H),2.58-2.53(m,2H),2.46-2.42(m,3H), 2.34-2.29(m,2H),2.18(s,3H),2.10-1.99(m,1H),1.51-1.42(m,4H),1.12-1.04(m,2H),1.04-0.96(m,2H).LCMS:[M+H] + = 877.6.

[0252] Example 17: Synthesis of N-(2-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)butyl)-3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoamine (dBPTF-23)

[0253] [ka]

[0254] 3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoic acid 3-amino-4-methoxybenzoic acid (5 g, 29.9 mmol) was suspended in acrylic acid (8.62 g, 119.6 mmol), and the resulting suspension was stirred at 100°C for 3 hours, then cooled to room temperature. Acetic acid (33 mL) was added, and the reaction mixture was stirred at 100°C for 10 minutes. Urea (11 g, 185.4 mmol) was added, and the reaction mixture was stirred at 120°C overnight. The reaction mixture was added to a solution of ice water (150 mL) and concentrated HCl aqueous solution (37%, 10 mL), and stirred overnight. The reaction mixture was filtered, the solid was washed with water, and dried. The solid was pulverized with HCl (0.05 M, 300 mL), filtered, and washed with tert-butyl methyl ether to obtain the title compound as a brown solid (6.4 g, 84%). LCMS:[M+H] + = 265.1.

[0255] Perfluorophenyl 3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoate In 5 mL of DCM, a solution of 3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoic acid (304 mg, 1.15 mmol), 2,3,4,5,6-pentafluorophenol (212 mg, 1.15 mmol), DMAP (329 mg, 1.73 mmol), and EDCI (14 mg, 0.12 mmol) was stirred at room temperature for 3 hours. The solution was diluted with water and extracted with ethyl acetate. The combined organic layers were dried over Na2SO4 and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using 60% ethyl acetate in PE to obtain the title compound as a white solid (299 mg, 60%). LCMS: [M+H] + = 431.0.

[0256] tert-butyl(2-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)ethyl)carbamate In 5 mL of DCE, a solution of 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (100 mg, 0.19 mmol) was added, to which DIEA (37 mg, 0.29 mmol) was added, and the reaction mixture was stirred at room temperature for 30 minutes. In 5 mL of DCE, a solution of tert-butyl(2-oxoethyl)carbamate (91 mg, 0.57 mmol) and STAB (122 mg, 0.57 mmol) was added, and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with water and extracted with DCM. The combined organic layers were dried over Na2SO4 and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using elution with 10% MeOH in DCM to obtain the title compound as a yellow solid (60 mg, 47%). LCMS:[M+H] + = 667.3.

[0257] 2-((4-((4-(2-aminoethyl)piperazin-1-yl)sulfonyl)-2-fluorophenyl)amino)-7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one To a solution of tert-butyl(2-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)ethyl)carbamate (50 mg, 0.07 mmol) in DCM (2 mL), HCl / dioxane (2 mL, 4 M) was added, and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a yellow solid (35 mg, crude), which was used without further purification. LCMS:[M+H] + = 567.2.

[0258] N-(2-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)ethyl)-3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzamide In a solution of 2-((4-((4-(2-aminoethyl)piperazin-1-yl)sulfonyl)-2-fluorophenyl)amino)-7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (35 mg, 0.06 mmol) in DCM (5 mL), DIEA (39 mg, 0.30 mmol) and perfluorophenyl 3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoate (15 mg, 0.05 mmol) were added, and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with water and extracted with DCM. The combined organic layer was concentrated to obtain a residue, which was purified by preparative HPLC to obtain the title compound as a white solid (33 mg, 67%). 1H NMR(400MHz,DMSO)δ10.34(s,1H),9.01(d,J=1.9Hz,1H),8.46(s,2H),8.29(t,J=5.6Hz,1H),8.07(dd,J=9.2,2.1Hz,1H),8.02-7 .95(m,2H),7.80(dd,J=8.6,2.3Hz,1H),7.73(d,J=2.2Hz,1H),7.61(dd,J=10.2,1.9Hz,1H),7.57-7.52(m,1H),7.39(d,J=9.2Hz, 1H),7.14(d,J=8.8Hz,1H),3.81(s,3H),3.80-3.74(m,1H),3.56(t,J=6.6Hz,2H),3.36-3.33(m,1H),3.31-3.28(m,1H),3.03-2.8 8(m,4H),2.70-2.62(m,2H),2.56-2.51(m,4H),2.48-2.43(m,2H),2.18(s,3H),1.12-1.05(m,2H),1.04-0.97(m,2H).LCMS:[M+H] + = 813.5.

[0259] Example 18: Synthesis of (2S,4R)-1-((S)-2-(11-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)undecaneamide)-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)pyrrolidine-2-formamide (dBPTF-24)

[0260] [ka]

[0261] Methyl 11-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)undecanoate In 2 mL of DMF, a solution of 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (100 mg, 0.19 mmol) was added, to which methyl 11-bromoundecanoate (53 mg, 0.19 mmol), KI (2 mg, 0.019 mmol), and NaHCO3 (24 mg, 0.28 mmol) were added. The reaction mixture was stirred in a microwave at 100°C for 1 hour. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using elution with 20% MeOH in DCM ether to obtain the title compound as a white solid (70 mg, 51%). LCMS:[M+H] + = 722.2.

[0262] 11-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)undecanoic acid In a solution of methyl 11-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazin-1-yl)undecanoate (70 mg, 0.097 mmol) in MeOH:THF:H2O (4:2:1) (3 mL), LiOH (9 mg, 0.388 mmol) was added, and the reaction mixture was stirred for 4 hours. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure to obtain the title compound as a white solid (50 mg, crude), which was used without further purification. LCMS:[M+H] + = 706.2.

[0263] (2S,4R)-1-((S)-2-(11-(4-((4-((7-(1-Cyclopropyl-1H-Pyrazole-4-yl)-3-Methyl-4-oxo-4H-Pyrido[1,2-a]Pyrimidine-2-yl)amino)-3-Fluorophenyl)Sulfonyl)Piperadin-1-yl)Undecanamide)-3,3-Dimethylbutanoyl)-4-Hydroxy-N-((S)-1-(4-(4-Methylthiazole-5-yl)phenyl)Ethyl)Pyrrolidine-2-Carboxamide In a solution of 11-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)undecanoic acid (50 mg, 0.071 mmol), (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (35 mg, 0.078 mmol), and DIEA (28 mg, 0.213 mmol), HATU (30 mg, 0.078 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with water (20 mL) and extracted with DCM (10 mL x 3). The combined organic layers were dried over Na2SO4 and concentrated to obtain a residue, which was purified by preparative HPLC (5% to 95% acetonitrile containing 0.1% FA in water) to obtain the title compound as a white solid (21 mg, 26%). 1H NMR(400MHz,DMSO)δ9.02(d,J=1.8Hz,1H),8.98(s,1H),8.47(s,2H),8.36(d,J=7.8Hz ,1H),8.09(dd,J=9.2,2.1Hz,1H),8.04-7.98(m,2H),7.76(d,J=9.3Hz,1H),7.61(dd,J =10.2,1.9Hz,1H),7.55(dd,J=8.5,1.9Hz,1H),7.45-7.43(m,1H),7.43-7.40(m,2H), 7.39-7.35(m,2H),5.08(d,J=3.5Hz,1H),4.94-4.87(m,1H),4.50(d,J=9.3Hz,1H),4.4 1(t,J=8.0Hz,1H),4.27(s,1H),3.81-3.73(m,1H),3.63-3.54(m,2H),2.95(s,4H),2. 45(d,J=2.5Hz,3H),2.42(s,4H),2.27-2.21(m,3H),2.18(s,3H),2.11-2.05(m,1H),2. 04-1.92(m,2H),1.82-1.75(m,1H),1.50-1.38(m,3H),1.36(d,J=7.0Hz,4H),1.23(s, 2H),1.20(s,9H),1.13-1.08(m,2H),1.02-0.98(m,2H),0.94-0.90(m,9H).LCMS:[M+H] + = 1134.4.

[0264] Example 19: Synthesis of N-(3-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)propyl)-3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoamine (dBPTF-25)

[0265] [ka]

[0266] 3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoic acid 3-amino-4-methoxybenzoic acid (5 g, 29.9 mmol) was suspended in acrylic acid (8.62 g, 119.6 mmol), and the resulting suspension was stirred at 100°C for 3 hours, then cooled to room temperature. Acetic acid (33 mL) was added, and the reaction mixture was stirred at 100°C for 10 minutes. Urea (11 g, 185.4 mmol) was added, and the reaction mixture was stirred at 120°C overnight. The reaction mixture was added to a solution of ice water (150 mL) and concentrated HCl aqueous solution (37%, 10 mL), and stirred overnight. The reaction mixture was filtered, the solid was washed with water, and dried. The solid was pulverized with HCl (0.05 M, 300 mL), filtered, and washed with tert-butyl methyl ether to obtain the title compound as a brown solid (6.4 g, 84%). LCMS:[M+H] + = 265.1.

[0267] Perfluorophenyl 3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoate In 5 mL of DCM, a solution of 3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoic acid (304 mg, 1.15 mmol), 2,3,4,5,6-pentafluorophenol (212 mg, 1.15 mmol), DMAP (329 mg, 1.73 mmol), and EDCI (14 mg, 0.12 mmol) was stirred at room temperature for 3 hours. The solution was diluted with water and extracted with ethyl acetate. The combined organic layers were dried over Na2SO4 and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using 60% ethyl acetate in PE to obtain the title compound as a white solid (299 mg, 60%). LCMS: [M+H] + = 431.0.

[0268] N-(2-(1,3-dioxolan-2-yl)ethyl)-3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzamide In a 5 mL solution of 2-(1,3-dioxolan-2-yl)ethane-1-amine, DIEA (207 mg, 1.6 mmol) was added, and the reaction mixture was stirred at room temperature for 30 minutes. Perfluorophenyl 3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzoate (135 mg, 0.32 mmol) was added, and the reaction mixture was stirred at room temperature overnight. The reaction mixture was diluted with water and extracted with DCM. The combined organic layers were dried over Na2SO4 and concentrated to obtain the residue, which was purified by silica gel column chromatography using elution with 10% MeOH in DCM to obtain the title compound as a white solid (100 mg, 86%). LCMS:[M+H] + =364.1.

[0269] 3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxy-N-(3-oxopropyl)benzamide In a solution of N-(2-(1,3-dioxolan-2-yl)ethyl)-3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzamide (20 mg, 0.06 mmol) in DCM (2 mL), TFA (2 mL) was added, and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a yellow solid (15 mg, crude), which was used without further purification. LCMS:[M+H] + =320.1.

[0270] N-(3-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)propyl)-3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxybenzamide In a solution of 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (21 mg, 0.04 mmol) in DCE (5 mL), DIEA (8 mg, 0.06 mmol) was added, and the reaction mixture was stirred at room temperature for 30 minutes. 3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-4-methoxy-N-(3-oxopropyl)benzoamide (15 mg, 0.05 mmol) and STAB (25 mg, 0.12 mmol) were added, and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with water and extracted with DCM. The combined organic layers were concentrated to obtain a residue, which was purified by preparative HPLC to obtain the title compound as a white solid (4 mg, 13%). 1 H NMR(400MHz,DMSO)δ10.35(s,1H),9.01(d,J=1.8Hz,1H),8.48(s,1H),8.46(s,1H),8.43(s,1H),8.38-8.27(m,1H),8.07(dd, J=9.2,2.0Hz,1H),8.04-7.98(m,2H),7.81(dd,J=8.7,2.1Hz,1H),7.76-7.73(m,1H),7.64-7.59(m,1H),7.59-7.53(m,1H),7 .41(d,J=9.2Hz,1H),7.15(d,J=8.7Hz,1H),3.84(s,3H),3.81-3.72(m,1H),3.57(t,J=6.4Hz,2H),3.24-3.17(m,4H),2.96(s ,4H),2.53-2.52(m,2H),2.48-2.42(m,4H),2.18(s,3H),1.67-1.58(m,2H),1.14-1.06(m,2H),1.03-0.96(m,2H).LCMS:[M+H] + = 827.6.

[0271] Example 20: Synthesis of (2S,4R)-1-((S)-2-(13-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)tridecaneamino)-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (dBPTF-26)

[0272] [ka]

[0273] Methyl 13-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)tridecanoate In 3 mL of DMF, a solution of 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (120 mg, 0.23 mmol) was added, to which methyl 13-bromotridecanoate (70 mg, 0.23 mmol), KI (2 mg, 0.023 mmol), and NaHCO3 (30 mg, 0.34 mmol) were added. The reaction mixture was stirred in a microwave at 100°C for 1 hour. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using elution with 20% MeOH in DCM ether to obtain the title compound as a white solid (110 mg, 64%). LCMS:[M+H] + = 750.3.

[0274] 13-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)tridecanoic acid In a solution of methyl 13-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazin-1-yl)tridecanoate (70 mg, 0.093 mmol) in MeOH:THF:H2O (4:2:1) (3 mL), LiOH (9 mg, 0.374 mmol) was added, and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure to obtain the title compound as a white solid (50 mg, crude), which was used without further purification. LCMS:[M+H] + = 736.2.

[0275] (2S,4R)-1-((S)-2-(13-(4-((4-((7-(1-Cyclopropyl-1H-Pyrazole-4-yl)-3-Methyl-4-oxo-4H-Pyrido[1,2-a]Pyrimidine-2-yl)amino)-3-Fluorophenyl)Sulfonyl)Piperadin-1-yl)Tridecanamino)-3,3-Dimethylbutanoyl)-4-Hydroxy-N-((S)-1-(4-(4-Methylthiazole-5-yl)phenyl)Ethyl)Pyrrolidine-2-Carboxamide In a solution of 13-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)tridecanoic acid (50 mg, 0.068 mmol), (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazole-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (33 mg, 0.074 mmol), and DIEA (26 mg, 0.204 mmol), HATU (28 mg, 0.074 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with water (20 mL) and extracted with DCM (10 mL x 3). The combined organic layers were dried over Na2SO4 and concentrated to obtain a residue, which was purified by preparative HPLC (5% to 95% acetonitrile containing 0.1% FA in water) to obtain the title compound as a white solid (15 mg, 18%). 1 H NMR(400MHz,DMSO)δ9.02(d,J=1.5Hz,1H),8.98(s,1H),8.47(s,2H),8.40 -8.33(m,1H),8.12-8.06(m,1H),8.05-7.98(m,2H),7.76(d,J=9.2Hz,1H) ,7.60(d,J=10.2Hz,1H),7.55(d,J=8.4Hz,1H),7.44-7.36(m,5H),5.25-5 .06(m,1H),4.94-4.78(m,1H),4.50(d,J=9.3Hz,1H),4.41(t,J=8.0Hz,1H ),4.27(s,1H),3.82-3.73(m,1H),3.64-3.55(m,2H),3.39(s,1H),2.95(s ,4H),2.45(s,3H),2.41(s,4H),2.27-2.21(m,3H),2.18(s,3H),2.11-1.9 7(m,2H),1.84-1.74(m,1H),1.51-1.41(m,2H),1.37(d,J=7.0Hz,4H),1.2 0(s,16H),1.12-1.07(m,2H),1.03-0.97(m,2H),0.92(s,9H).LCMS:[M+H] + = 1162.4.

[0276] Example 21: Synthesis of 6-(3-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)propyl)-2-(2,6-dioxopiperidine-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione (dBPTF-27)

[0277] [ka]

[0278] 2-((4-((4-(2-(1,3-dioxolan-2-yl)ethyl)piperazine-1-yl)sulfonyl)-2-fluorophenyl)amino)-7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one In 3 mL of DMF, a solution of 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyridino[1,2-a]pyrimidine-4-one (140 mg, 0.27 mmol) was added to DIEA (139 mg, 1.08 mmol), 2-(2-bromoethyl)-1,3-dioxolane (147 mg, 0.81 mmol), and KI (49 mg). The reaction mixture was stirred in a microwave at 80°C for 1 hour. This reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by silica gel column elution with 10% MeOH in DCM to obtain the title compound as a yellow solid (60 mg, 36%). LCMS: [M+H] + = 624.0.

[0279] 3-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)propanal A solution of 2-((4-((4-(2-(1,3-dioxolan-2-yl)ethyl)piperazine-1-yl)sulfonyl)-2-fluorophenyl)amino)-7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (60 mg, 0.1 mmol) in formic acid (2 mL) was stirred at 50°C for 10 hours. The reaction mixture was concentrated to obtain a residue, which was purified by silica gel column elution with 5% MeOH in DCM to obtain the title compound as a white solid (15 mg, 26%). LCMS:[M+H] + = 580.1.

[0280] 6-(3-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)propyl)-2-(2,6-dioxopiperidine-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione In a solution of 2-(2,6-dioxopiperidine-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione (8 mg, 0.02 mmol) in DCM (5 mL), DIEA (5 mg, 0.04 mmol) was added, and the reaction mixture was stirred at room temperature for 30 minutes. 3-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)propanal (15 mg, 0.02 mmol) and STAB (17 mg, 0.08 mmol) were added, and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with water and extracted with DCM. The combined organic layers were concentrated to obtain a residue, which was purified by preparative HPLC to obtain the title compound as a white solid (3 mg, 7%). 1 H NMR(400MHz,DMSO)δ11.11(s,1H),9.01(d,1H),8.53-8.48(m,1H),8.48-8.46(m,1H),8.13-8.13(m,1H),8.10-8.06(m,1H),8.05 -7.97(m,2H),7.74(s,2H),7.66-7.60(m,1H),7.58-7.54(m,1H),7.42(d,J=9.1Hz,1H),5.12(dd,J=12.9,5.2Hz,1H),3.91(s,4H) ,3.80-3.73(m,1H),3.05-2.91(m,4H),2.87(d,J=12.4Hz,1H),2.67-2.62(m,2H),2.58-2.54(m,1H),2.47-2.43(m,3H),2.40-2.3 2(m,2H),2.19(s,3H),2.08-1.97(m,2H),1.71-1.54(m,2H),1.52-1.40(m,1H),1.15-1.05(m,2H),1.05-0.94(m,2H).LCMS:[M+H] + = 863.6.

[0281] Example 22: Synthesis of 6-(6-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)hexyl)-2-(2,6-dioxopiperidine-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione (dBPTF-28)

[0282] [ka]

[0283] 6-(5-(1,3-dioxolan-2-yl)pentyl)-2-(2,6-dioxopiperidine-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione In 2 mL of ACN, a solution of 2-(2,6-dioxopiperidine-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione (100 mg, 0.33 mmol) was mixed with DIEA (129 mg, 1.00 mmol), 2-(5-bromopentyl)-1,3-dioxolane (82.1 mg, 0.37 mmol), and KI (61.1 mg, 0.37 mmol). The reaction mixture was stirred in a microwave at 80°C for 1 hour. The reaction mixture was concentrated to obtain a residue, which was purified by flash silica gel chromatography (10% MeOH in DCM) to obtain the title compound as a yellow solid (78 mg, 35%). LCMS: [M+H] + = 442.3.

[0284] 6-(6-(2,6-dioxopiperidine-3-yl)-5,7-dioxo-3,5,6,7-tetrahydropyrrolo[3,4-f]isoindole-2(1H)-yl)hexanal In a solution of 6-(5-(1,3-dioxolan-2-yl)pentyl)-2-(2,6-dioxopiperidine-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione (50 mg, 0.11 mmol) in DCM (2 mL), TFA (2 mL) was added, and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a yellow solid (40 mg, crude), which was used without further purification. LCMS: [M+H] + =398.1.

[0285] 6-(6-(4-((4-((7-(1-cyclopropyl-1H-pyrazole-4-yl)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-2-yl)amino)-3-fluorophenyl)sulfonyl)piperazine-1-yl)hexyl)-2-(2,6-dioxopiperidine-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione In a 5 mL solution of DCE containing 7-(1-cyclopropyl-1H-pyrazole-4-yl)-2-((2-fluoro-4-(piperazine-1-ylsulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (53 mg, 0.10 mmol), DIEA (19 mg, 0.15 mmol) was added, and the reaction mixture was stirred at room temperature for 30 minutes. 6-(6-(2,6-dioxopiperidine-3-yl)-5,7-dioxo-3,5,6,7-tetrahydropyrrolo[3,4-f]isoindole-2(1H)-yl)hexanal (40 mg, 0.10 mmol) and STAB (64 mg, 0.30 mmol) were added, and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with water and extracted with DCM. The combined organic layers were concentrated to obtain a residue, which was purified by preparative HPLC to obtain the title compound as a white solid (14 mg, 9%). 1H NMR(400MHz,DMSO)δ11.11(s,1H),8.99(d,J=1.6Hz,1H),8.51-8.42(m,2H),8.10-8.05(m,1H),8.04-7.98(m,2H),7.72(s,2H),7. 61(dd,J=10.2,1.8Hz,1H),7.55(d,J=8.6Hz,1H),7.39(d,J=9.2Hz,1H),5.12(dd,J=12.9,5.3Hz,1H),3.91(s,4H),3.80-3.73(m, 1H),2.94(d,J=13.1Hz,4H),2.91-2.83(m,1H),2.68-2.58(m,3H),2.58-2.53(m,1H),2.44(s,4H),2.28(t,J=6.2Hz,2H),2.17(s, 3H),2.11-2.02(m,1H),1.52-1.43(m,2H),1.43-1.35(m,2H),1.35-1.19(m,5H),1.13-1.05(m,2H),1.05-0.96(m,2H).LCMS:[M+H] + =905.4.

[0286] Example 23: Synthesis of 5-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-5-yl)piperazine-1-yl)-N-(2-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)ethyl)pentanamide (dBPTF-29)

[0287] [ka]

[0288] The title compound was prepared according to a similar procedure to that described in the above examples. LCMS:[M+H] + =965.5.

[0289] Example 24: Synthesis of 7-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-5-yl)piperazine-1-yl)-N-(2-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)ethyl)heptanamide (dBPTF-30)

[0290] [ka]

[0291] The title compound was prepared according to a similar procedure to that described in the above examples. LCMS:[M+H] + = 993.6.

[0292] Example 25: Synthesis of 2-(4-(4-((2,6-dioxopiperidine-3-yl)amino)-2-fluorophenyl)piperazine-1-yl)-N-(2-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)ethyl)acetamide (dBPTF-31)

[0293] [ka]

[0294] The title compound was prepared according to a similar procedure to that described in the above examples. LCMS:[M+H] + = 887.5.

[0295] Example 26: Synthesis of 11-(2-(2,6-dioxopiperidine-3-yl)-1-oxoisoindorin-4-yl)-N-(2-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)ethyl)undeca10amide (dBPTF-32)

[0296] [ka]

[0297] The title compound was prepared according to a similar procedure to that described in the above examples. LCMS:[M+H] + =947.6.

[0298] Example 27: Synthesis of N1-(2-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)ethyl)-N8-((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)octanediamide (dBPTF-33)

[0299] [ka]

[0300] The title compound was prepared according to a similar procedure to that described in the above examples. LCMS:[M+H] + = 1123.6.

[0301] Example 28: Synthesis of (2S,4R)-N-(2-((7-((2-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)ethyl)amino)-7-oxoheptyl)oxy)-4-(4-methylthiazole-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamide)-3,3-dimethylbutanol)-4-hydroxypyrrolidine-2-carboxamide (dBPTF-34)

[0302] [ka]

[0303] The title compound was prepared according to a similar procedure to that described in the above examples. LCMS:[M+H] + = 1183.8.

[0304] Example 29: Synthesis of (2S,4R)-N-(2-((7-((2-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)ethyl)amino)-7-oxoheptyl)oxy)-4-(4-methylthiazole-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamide)-3,3-dimethylbutanol)-4-hydroxypyrrolidine-2-carboxamide (dBPTF-38)

[0305] [ka]

[0306] 2-(2,6-dioxopiperidine-3-yl)-5-(piperazine-1-yl)isoindorin-1,3-dione In a solution of tert-butyl 4-2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-5-yl)piperazine-1-carboxylate (200 mg, 0.45 mmol) in DCM (2 mL), 4 M HCl was added in a dioxane solution (1 mL), and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a yellow solid (200 mg, crude), which was used without further purification. LCMS: [M+H] + = 343.1.

[0307] 5-(4-(3-(1,3-dioxolan-2-yl)propyl)piperazine-1-yl)-2-(2,6-dioxopiperidine-3-yl)isoindorin-1,3-dione In 20 mL of MeCN, a solution of 2-(2,6-dioxopiperidine-3-yl)-5-(piperazine-1-yl)isoindoline-1,3-dione (200 mg, 0.58 mmol) was mixed with 2-(3-bromopropyl)-1,3-dioxolane (226 mg, 1.17 mmol), K2CO3 (242 mg, 1.75 mmol), and KI (106 mg, 0.64 mmol). The reaction mixture was stirred at 75°C for 3 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 mL x 2), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using elution with 5% MeOH in DCM to obtain the title compound as a yellow solid (200 mg, 75%). LCMS: [M+H] + = 457.3.

[0308] 4-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-5-yl)piperazine-1-yl)butanal In a solution of 5-(4-(3-(1,3-dioxolan-2-yl)propyl)piperazin-1-yl)-2-(2,6-dioxopiperidine-3-yl)isoindoline-1,3-dione (50 mg, 0.11 mmol) in DCM (1.5 mL), TFA (1.5 mL) was added, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a yellow solid (400 mg, crude), which was used without further purification. LCMS: [M+H] + = 413.1.

[0309] 7-(1-(azetidine-3-yl)-1H-pyrazole-4-yl)-2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one In a solution of tert-butyl 3-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)azetidine-1-carboxylate (45 mg, 0.069 mmol) in DCM (2 mL), TFA (0.5 mL) was added, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a white solid (45 mg, crude), which was used without further purification. LCMS:[M+H] + = 553.1.

[0310] 2-(2,6-dioxopiperidine-3-yl)-5-(4-(4-(3-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)azetidine-1-yl)butyl)piperazine-1-yl)isoindoline-1,3-dione DCE (6 mL) contains 7-(1-(azetidine-3-yl)-1H-pyrazole-4-yl)-2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (31 mg, 0.048 mmol) and 4-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-5-yl)piperazine-1-yl)butanal (20 mg, 0.048 mmol). To a solution of 0.19 mmol (41.16 mg) of STAB (0.19 mmol), the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with water (100 mL) and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (30 mL x 2), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by preparative HPLC (5% to 95% acetonitrile containing 0.1% NH3·H2O in water) to obtain the title compound as a yellow solid (4 mg, 7%). 1 H NMR(400MHz,DMSO)δ11.08(s,1H),9.07-9.03(m,1H),8.60(s,1H),8.50(s,1H),8.14(s,1H),8.13-8.08(m,2H),8.03-7.97 (m,1H),7.67(d,J=8.5Hz,1H),7.62(dd,J=10.2,1.7Hz,1H),7.55(dd,1H),7.43(d,J=9.2Hz,1H),7.34(s,1H),7.26(dd,J= 8.6Hz,1H),5.10-5.00(m,2H),3.85-3.76(m,2H),3.51-3.42(m,9H),2.97(s,4H),2.91-2.82(m,1H),2.64-2.54(m,4H),2. 52(s,1H),2.42-2.32(m,6H),2.18(d,J=10.1Hz,6H),2.06-1.95(m,1H),1.56-1.45(m,2H),1.43-1.34(m,2H).LCMS:[M+H] + =949.3.

[0311] Example 30: Synthesis of 2-(2,6-dioxopiperidine-3-yl)-4-(4-(8-(3-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)azetidine-1-yl)octyl)piperazine-1-yl)isoindoline-1,3-dione (dBPTF-39)

[0312] [ka]

[0313] 1-((3-fluoro-4-nitrophenyl)sulfonyl)-4-methylpiperazine In a 20 mL solution of 1-methylpiperazine (836 mg, 8.34 mmol) in DCM, TEA (2.5 g, 25.04 mmol) and 3-fluoro-4-nitrobenzenesulfonyl chloride (2 g, 8.34 mmol) were added at 0°C. The reaction mixture was stirred at 0°C for 0.3 hours. The reaction mixture was concentrated to obtain a residue, which was purified by silica gel column elution with 40% MeOH in DCM to obtain the title compound as a yellow solid (2.3 g, 91%). LCMS: [M+H] + =304.0.

[0314] 2-Fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)aniline In 20 mL of EtOH, a solution of 1-((3-fluoro-4-nitrophenyl)sulfonyl)-4-methylpiperazine (2.3 g, 7.59 mmol) was added, and Pd / C (230 mg) was passed through. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was filtered through a Celite® pad and washed with DCM. The filtrate was concentrated under reduced pressure to obtain the title compound as a yellow solid (2 g, crude). LCMS: [M+H] + =274.0.

[0315] tert-butyl3-(4-(2-chloro-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)azetidine-1-carboxylate To a solution of 7-bromo-2-chloro-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (200 mg, 0.732 mmol) in dioxane (20 mL) and H2O (5 mL), tert-butyl 3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-1-yl)azetidine-1-carboxylate (255 mg, 0.73 mmol), Na2CO3 (155 mg, 1.46 mmol), and Pd(dppf)Cl2 (53 mg, 0.073 mmol) were added, and the reaction mixture was stirred at 100°C for 3 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 mL x 2), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using 60% EA in PE to obtain the title compound as a yellow solid (200 mg, 66%). LCMS: [M+H] + =360.0.

[0316] tert-butyl3-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)azetidine-1-carboxylate To a solution of tert-butyl 3-4-(2-chloro-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)azetidine-1-carboxylate (200 mg, 0.48 mmol) in toluene (1.5 mL), 2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)aniline (157 mg, 0.57 mmol), RuPhospdG3 (40 mg, 0.048 mmol), and Cs2CO3 (314 mg, 0.96 mmol) were added, and the reaction mixture was stirred at 110°C for 3 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 mL x 2), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using elution with 5% MeOH in DCM to obtain the title compound as a pink solid (200 mg, 60%). LCMS:[M+H] + = 653.6.

[0317] 7-(1-(azetidine-3-yl)-1H-pyrazole-4-yl)-2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one In a solution of tert-butyl 3-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)azetidine-1-carboxylate (45 mg, 0.069 mmol) in DCM (2 mL), TFA (0.5 mL) was added, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a white solid (45 mg, crude), which was used without further purification. LCMS:[M+H] + = 553.5.

[0318] 2-(2,6-dioxopiperidine-3-yl)-4-(4-(8-(3-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)azetidine-1-yl)octyl)piperazine-1-yl)isoindoline-1,3-dione In a solution of 7-(1-(azetidine-3-yl)-1H-pyrazole-4-yl)-2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (40 mg, 0.072 mmol) and 8-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-4-yl)piperazine-1-yl)octanal (30 mg, 0.072 mmol), STAB (61 mg, 0.28 mmol) was added, and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with water (100 mL) and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (30 mL x 2), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by preparative HPLC (5% to 95% acetonitrile containing 0.1% NH3·H2O in water) to obtain the title compound as a yellow solid (5 mg, 7%). 1H NMR(400MHz,DMSO)δ11.09(s,1H),9.05(d,J=1.8Hz,1H),8.60(s,1H),8.52(s,1H),8.17(s,1H),8.14(s,1H),8.10(dd,J=9. 2,2.0Hz,1H),8.02-7.95(m,1H),7.73-7.67(m,1H),7.62(dd,J=10.2,1.9Hz,1H),7.56(dd,J=8.5,1.8Hz,1H),7.43(d,J=9. 2Hz,1H),7.39-7.32(m,2H),5.19-5.05(m,2H),3.91(d,J=106.8Hz,4H),2.97(s,4H),2.93-2.68(m,6H),2.59(d,J=18.4Hz, 2H),2.53(d,J=5.1Hz,3H),2.41(s,7H),2.19(s,6H),2.06-1.98(m,1H),1.51(s,2H),1.40(s,2H),1.27(s,9H).LCMS:[M+H] + = 1005.5.

[0319] Example 31: Synthesis of 2-(2,6-dioxopiperidine-3-yl)-5-(4-(6-(3-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)azetidine-1-yl)hexyl)piperazine-1-yl)isoindoline-1,3-dione (dBPTF-40)

[0320] [ka]

[0321] 5-(4-(5-(1,3-dioxolan-2-yl)pentyl)piperazine-1-yl)-2-(2,6-dioxopiperidine-3-yl)isoindorine-1,3-dione In 20 mL of ACN, a solution of 2-(2,6-dioxopiperidine-3-yl)-5-(piperazine-1-yl)isoindoline-1,3-dione (200 mg, 0.58 mmol) was added to 2-(5-bromopentyl)-1,3-dioxolane (260 mg, 1.17 mmol), DIEA (301 mg, 2.34 mmol), and KI (106 mg, 0.64 mmol). The reaction mixture was stirred at 80°C for 16 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (30 mL x 2), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by silica gel column chromatography using elution with 3% MeOH in DCM to obtain the title compound as a yellow solid (160 mg, 87%). LCMS: [M+H] + = 485.3.

[0322] 6-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-5-yl)piperazine-1-yl)hexanal To a solution of 5-(4-(5-(1,3-dioxolan-2-yl)pentyl)piperazin-1-yl)-2-(2,6-dioxopiperidine-3-yl)isoindoline-1,3-dione (50 mg, 0.11 mmol) in DCM (1 mL), TFA (1.5 mL) was added, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain the title compound as a yellow solid (400 mg, crude), which was used without further purification. LCMS: [M+H] + = 441.1.

[0323] 2-(2,6-dioxopiperidine-3-yl)-5-(4-(6-(3-(4-(2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-7-yl)-1H-pyrazole-1-yl)azetidine-1-yl)hexyl)piperazine-1-yl)isoindoline-1,3-dione In a solution of 7-(1-(azetidine-3-yl)-1H-pyrazole-4-yl)-2-((2-fluoro-4-((4-methylpiperazine-1-yl)sulfonyl)phenyl)amino)-3-methyl-4H-pyrido[1,2-a]pyrimidine-4-one (40 mg, 0.072 mmol) and 6-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-5-yl)piperazine-1-yl)hexanal (31 mg, 0.072 mmol), STAB (61.44 mg, 0.29 mmol) was added, and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with water (100 mL) and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (30 mL x 2), dried over Na2SO4, and concentrated to obtain a residue. This residue was purified by preparative HPLC (5% to 95% acetonitrile containing 0.1% NH3·H2O in water) to obtain the title compound as a yellow solid (5 mg, 7%). 1 H NMR(400MHz,DMSO)δ11.07(s,1H),9.03(d,J=1.7Hz,1H),8.59(s,1H),8.48(s,1H),8.15(s,1H),8.12-8.07(m,2H),8.03-7.97(m,1H),7.68 -7.64(m,1H),7.61(dd,J=10.2,1.9Hz,1H),7.55(dd,J=8.5,1.8Hz,1H),7.42(d,J=9.2Hz,1H),7.32(d,J=1.8Hz,1H),7.24(dd,J=8.6,2.0H) z,1H),5.10-4.94(m,2H),3.76-3.68(m,2H),3.41(d,J=5.3Hz,6H),3.03-2.82(m,5H),2.58(dd,J=19.0,4.5Hz,2H),2.53-2.51(m,2H),2.4 9-2.46(m,4H),2.41-2.36(m,4H),2.34-2.28(m,2H),2.17(d,J=10.9Hz,6H),2.06-1.95(m,1H),1.52-1.41(m,2H),1.32(s,6H).LCMS:[M+H] + =977.3.

[0324] Example 32: Biological Data Cell viability assay The CellTiter-Glo® assay was performed according to the manufacturer's instructions (Promega®). Briefly, 1,000 cells per well were seeded in a 96-well plate and treated with a range of compound concentrations. Cell viability was measured at the stated time points based on luminescence obtained by the CellTiter-Glo® assay (Promega®) and read using an EnVision® 2104 (PerkinElmer) according to the manufacturer's protocol.

[0325] Imbubrot Cells growing in culture were lysed for whole cell lysates. Nuclear lysates were prepared using the NE-PER® nuclear lysate kit (Thermo Fisher Scientific®) according to the manufacturer's protocol. Chromatin lysates were prepared using the whole histone extraction kit (Epigentek). Briefly, equal volumes of proteins were separated by Western blotting using 4%–12% Bis-Tris NuPAGE® gel (Thermo Fisher Scientific®), transcribed, and immunoblotted with primary antibodies against BPTF, PARP, cleaved PARP, β-actin, GSPT1, and IKZF1. The secondary antibody was IRDye® conjugate antibody (Licor).

[0326] BPTF is a member of the chromatin remodeling complex (NURF) that binds to chromatin and adds covalent modifications to histone proteins, influencing downstream transcription. Compounds of formula I or II (e.g., NVS-BPTF-1B / dBPTF-2) remove BPTF from histones, while NVS-BPTF-1 does not. BPTF has two contact sites with histones, and NVS-BPTF-1 can target only one of them, while compounds of formula I or II can target both simultaneously in a short time (Figure 10).

[0327] Complete removal of BPTF from histones was achieved by 8 hours, but the presence of BPTF in the nucleus at 24 hours indicated that it was not completely degraded. Removing the entire protein from DNA interrupted its disease-promoting function, even without complete proteolysis. BPTF is a component of the NURF chromatin remodeling complex, and the entire complex is a member that interacts with DNA. Complete removal of BPTF from histones destabilizes NURF as a whole (Figures 11A-11B).

[0328] Colony formation assay Colony assays were performed by reseeding cells in 6-well dishes with 500 cells per well, growing them in standard growth medium for 10 days, followed by 100% methanol fixation, 0.05% crystal violet staining, and subsequent quantification.

[0329] Lentivirus infection (shRNA) ShRNA targeting BPTF was ordered as a bacterial stock and transformed into competent E. coli cells. Bacterial DNA was collected and isolated to generate lentiviral particles produced in 293T cells. Plasmids were transfected into 293T cells using Lipofectamine® 2000 (Invitrogen®) along with pMD2.G (Addgene_12259) and psPAX2 (Addgene_12260) to generate viral particles using standard methodology. Stable Cas9-expressing RH4 cells were infected with BPTF lentiviral particles and selected with puromycin. Knockdown efficiency was established by immunoblotting.

[0330] Animal testing For tumor studies, 6-8 week old female NSG mice (The Jackson Laboratory) were subcutaneously transplanted with 2 × 10⁶ RH4 cells in 50% Matrigel / PBS. The mice were assigned to two groups: vehicle (n=15) or dBPTF-9 (50 mg / kg / day, n=15) administered via intravenous injection. Tumors engrafted and reached 100 mm. 3 Treatment with small molecule inhibitors was initiated when the target was reached. Mice were treated until the endpoint was reached, and their survival was tracked. Tumors were measured with a caliper, and mice were weighed every three days. If the tumor reached 2,000 mm in length or width, or if the animal became mortally ill, the animal was euthanized according to facility guidelines.

[0331] Example 33: BPTF assay Hela cell lines were manipulated using the HiBit system, and changes in protein levels were reported. Cell lines were treated with compounds, and fluorescence signals were measured at the indicated time points (6 hours or 20 hours). A single decrease in dose-response mode was observed, and DCs were also identified. 50 It was used for measurement.

[0332] [Table 1] NA - Unavailable (not observed due to low activity or lack of degradation).

[0333] All patent and non-patent publications represent the level of skill of a person skilled in the art to which this disclosure pertains. All of these publications (including any specific portion thereof referenced) are incorporated herein by reference to the same extent as each individual publication is specifically and individually indicated as being incorporated by reference.

[0334] While the disclosures herein are described with reference to specific embodiments, it should be understood that these embodiments are merely illustrative of the principles and uses of the disclosure. Therefore, it should be understood that numerous modifications may be made to the exemplary embodiments and other configurations may be devised without departing from the spirit and scope of the disclosure as defined by the appended claims.

Claims

1. A compound having a structure represented by formula I or II: 【Chemistry 1】 During the ceremony, Degron binds to cerebron (CRBN) or von Hippel-Lindau (VHL) tumor suppressors. A compound in which the linker represents the part that covalently bonds degron with a nitrogen atom, or a pharmaceutically acceptable salt or stereoisomer thereof.

2. Degron is represented by a structure selected from the following group: 【Chemistry 2】 During the ceremony, X is CH ２ , or C(O), R １ However, it does not exist, CH ２ , 【Transformation 3】 A 5- to 11-membered monocyclic or bicyclic heterocycline containing one or two heteroatoms independently selected from NH, O, or N, O, and S, X' is either absent or C(O), Y is N or CH, R ２ The compound according to claim 1, wherein the substituent is optional.

3. The compound according to claim 2, wherein degron is represented by formula D1.

4. X is CH ２ The compound according to claim 3.

5. The compound according to claim 3, wherein X is C(O).

6. The compound according to claim 2, wherein degron is of formula D2.

7. X' is nonexistent, R ２ The compound according to claim 6, wherein it is fluoro.

8. R １ The compound according to claim 2, wherein the ion is absent or NH.

9. R １ The compound according to claim 2, wherein the compound is a six-membered heterocycline containing one or two heteroatoms independently selected from N, O, and S.

10. R １ The compound according to claim 9, which is piperazinyl.

11. R １ The compound according to claim 2, wherein the compound is a nine-membered bicyclic heterocycline containing one or two heteroatoms independently selected from N, O, and S.

12. R １ The compound according to claim 11, wherein the compound is 2,7-diazaspiro[3.5]nonane.

13. The compound according to claim 2, wherein degron is represented by formula D4.

14. R ２ The compound according to claim 13, wherein the compound is methoxy.

15. The compound according to claim 2, wherein degron is represented by formula D5.

16. Deglon, D6: 【Chemistry 4】 or represented by its stereoisomer, During the ceremony, R ３ However, hydrogen, or optionally substituted C １ ~C ３ Alkyl, or R ３ , and R ４ However, together with the carbon atoms to which they are bonded, they form cyclopropyl, R ４ However, hydrogen, methyl, or 【Transformation 5】 And, R ５ Yes, C (O) C ６ R ７ R ８ 、 【Transformation 6】 And, R ６ , and R ７ However, it is hydrogen, or R ６ , and R ７ However, together with the carbon atoms to which they are bonded, they form cyclopropyl, R ８ However, hydrogen, fluoro, cyano, or NMee ２ And, Y is hydrogen, 【Transformation 7】 And, 【Transformation 8】 The compound according to claim 1, wherein the bond is between degron and linker, and there is only one bond between degron and linker.

17. A degron is represented by one of the following structures: 【Chemistry 9】 or its stereoisomer, In the formula, Y is O, NH, or CH ２ The compound according to claim 16.

18. The linker is -O-, -S-, -N(R')-, 【Chemistry 10】 -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(NOR')-, -C(O)N(R')-, -C(O)N(R')C(O)-, -C(O)N(R')C(O)N(R')-, -N(R')C(O )-, -N(R')C(O)O-, -OC(O)N(R')-, -C(NR')-, -N(R')C(NR')-, -C(NR')N(R')-, -N(R')C(NR')N(R')-, -OB(Me)O-, -S(O) ２ -, -OS(O)-, -S(O)O-, -S(O)-, -OS(O) ２ -, -S(O) ２ O-, -N(R')S(O) ２ -, -S(O) ２ N(R')-, -N(R')S(O)-, -S(O)N(R')-, -N(R')S(O) ２ N(R')-, -N(R')S(O)N(R')-, and C as arbitrarily substituted. ３ ~C １２ An alkylene chain that may be interrupted and / or terminated at either or both ends of at least one of the following: a carbocyclene, an optionally substituted 3- to 12-membered heterocyclene, an optionally substituted 5- to 12-membered heteroarylene, or any combination thereof, wherein R' is H or C １ ~C ６ The compound according to claim 1, wherein the alkyl group and one or both terminal groups may be the same or different.

19. The compound according to claim 18, wherein the linker is an alkylene chain having 3 to 11 alkylene units.

20. Linker, 【Chemistry 11】 【Chemistry 12】 The compound according to claim 18. 【Request Item 21】 【Chemistry 13】 【Chemistry 14】 【Chemistry 15】 【Chemistry 16】 【Chemistry 17】 [Chemistry 18] 【Chemistry 19】 【Chemistry 20】 【Chemistry 21】 The compound according to claim 1, or a pharmaceutically acceptable salt or stereoisomer thereof. 【Request Item 22】 【Chemistry 22】 It is a compound.

23. A pharmaceutical composition comprising a compound according to any one of claims 1 to 21, or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier.

24. A method for treating cancer accompanied by abnormal activity of bromodomain PHD finger transcription factor (BPTF), comprising administering to a subject as needed a compound according to any one of claims 1 to 21, or a pharmaceutically acceptable salt or stereoisomer thereof, or the pharmaceutical composition according to claim 23.

25. The method according to claim 24, wherein the cancer is a sarcoma.

26. The method according to claim 25, wherein the sarcoma is rhabdomyosarcoma.

27. The method according to claim 24, wherein the cancer is breast cancer, lung cancer, or lymphoma.

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