CN119060067A

CN119060067A - Intermediates and methods for compounds that degrade EGFR kinase

Info

Publication number: CN119060067A
Application number: CN202410696695.5A
Authority: CN
Inventors: 雷柏林; 刘华庆; 赵逸舟; 亓新柱; 刘冠军; 靳琛; 尹建新; 王志伟
Original assignee: Baiji Shenzhou Beijing Biotechnology Co ltd
Current assignee: Baiji Shenzhou Beijing Biotechnology Co ltd
Priority date: 2023-05-31
Filing date: 2024-05-31
Publication date: 2024-12-03

Abstract

Disclosed herein are novel bifunctional compounds formed by conjugating an EGFR inhibitor moiety with an E3 ligase ligand moiety that function to recruit a target protein to the E3 ubiquitin ligase for degradation, and methods of making and using the same.

Description

Intermediates and methods for compounds that degrade EGFR kinase

Technical Field

Background

Proteolytically targeted chimeras (PROTAC) consist of two covalently linked protein binding molecules, one capable of binding to E3 ubiquitin ligase and the other to the target Protein (POI), a target that is intended for degradation (Sakamoto KM et al, proc. Natl. Acad. Sci.2001,98:8554-9.; sakamoto K. M. Et al, methods enzymol.2005; 399:833-847.). Recruiting E3 ligase to a specific unwanted protein rather than inhibiting the enzymatic activity of the target protein results in ubiquitination of the target protein and subsequent degradation of the target protein by the proteasome. The entire process of ubiquitination and proteasome degradation is known as the ubiquitin-proteasome pathway (UPP) (Ardley H. Et al, essays biochem.2005,41,15-30; komander D. Et al, biochem.2012,81,203-229; grice G.L. Et al, cell Rep.2015,12,545-553; swatek K.N. Et al, cell Res.2016,26, 399-422). Proteasomes are protein complexes that degrade unwanted, misfolded or abnormal proteins into small peptides to maintain cell health and productivity. Ubiquitin ligases (also known as E3 ubiquitin ligases) directly catalyze the transfer of ubiquitin from E2 to target proteins for degradation. Although the human genome encodes more than 600 putative E3 ligases, only a limited number of E3 ubiquitin ligases are widely used by the small molecule PROTAC technology, cereblon (CRBN), von Hippel-Lindau (VHL), mouse double-minute 2 homolog (MDM 2) and apoptosis inhibitor protein (cIAP) (Philipp O. Et al, chem.biol.2017,12, 2570-2578), recombinant human ring finger protein 114 (RNF 114) (Spradlin, J.N. Et al, nat.chem.biol.2019,15, 747-755), and DDB1 and CUL4 related factor 16 (DCAF 16) (Zhang, X. Et al, nat.chem.biol.2019,15, 737-746). For example, cereblon (CRBN) forms an E3 ubiquitin ligase complex with damaged DNA binding protein 1 (DDB 1) and Cullin-4A (CUL 4A) to ubiquitinate many other proteins, which are then degraded via the proteasome. (Yi-An Chen et al SCIENTIFIC REPORTS, 5, 1-13). Immunomodulatory Drugs (IMiD), including thalidomide (thalidomide), lenalidomide (lenalidomide), and pomalidomide (pomalidomide), act as monovalent promoters of PPI by binding to the Cereblon (CRBN) subunit of the CRL4a ^CRBN E3 ligase complex and recruiting new substrate proteins. (MATYSKIELA, M.E. et al, nat Chem Biol 2018,14,981-987). Thus, the ability of thalidomide and its derivatives to recruit CRBN has been widely used in proteolytically targeted chimeric (PROTAC) related studies (Christopher t. Et al ACS chem. Biol.2019,14, 342-347; honorine l. Et al ACS cent. Sci.2016,2, 927-934). PROTAC have great potential to eliminate the traditional inhibitor "non-druggable (undruggable)" or as a protein target for non-enzymatic proteins. (Chu TT. et al, cell Chem biol.2016;23:453-461.Qin C. Et al, J Med Chem 2018;61:6685-6704.Winter GE. Et al, science 2015; 348:1376-1381). In recent years PROTAC has been reported in antitumor studies as a useful modulator to promote selective degradation of a variety of target proteins. (Lu J. Et al, chem biol.2015;22 (6): 755-763; ottis P. Et al, chem biol.2017;12 (4): 892-898.; crews C.M. Et al, J Med chem.2018;61 (2): 403-404; neklesa T.K. Et al, pharmacol Ther.2017,174:138-144.; cermakova K. Et al Molecules,2018.23 (8); an S. Et al, EBioMedicine, lebraud H. Et al, essays biochem.2017;61 (5): 517-527.; sun Y.H. Et al, cell Res.2018;28:779-81; toure M et al, ANGEW CHEM INT ED Engl.2016;55 (6: 1966-1973; ykeghun et al, leu.S. 25, pages 2020 and publications such as those described in pages 802, 220 and 35, pages 162, etc., pages 802, 220 and 220, respectively, and publications such as those described in WO patent publications, 220, and publications.

Epidermal Growth Factor Receptor (EGFR), which belongs to the ErbB family, is a transmembrane Receptor Tyrosine Kinase (RTK) that plays a fundamental key role in cell proliferation, differentiation and motility (Y. Yarden et al, nat. Rev. Mol. Cell biol.2001; 2:127-137.). Homodimerization or heterodimerization of EGFR and other ErbB family members activates cytoplasmic tyrosine kinase domains to initiate intracellular signaling. Overexpression or activation mutations in EGFR have been associated with the development of many types of cancers, such as pancreatic cancer, breast cancer, glioblastoma multiforme, head and neck cancer, and non-small cell lung cancer (Yewale C. Et al, biomaterials.2013,34 (34): 8690-8707.). Activating mutations in the EGFR tyrosine kinase domain (L858R mutation and exon 19 deletion) have been identified as oncogenic drivers of NSCLC (Konduri, K. Et al, cancer Discovery 2016,6 (6), 601-611.). First generation EGFR tyrosine kinase inhibitors (EGFR-TKI) gefitinib and erlotinib have been approved for use in NSCLC patients with EGFR activating mutations (M.Maemondo, N.Engl.J.Med.362 (2010) 2380-2388.). While most patients with EGFR mutant NSCLC respond to these treatments, patients often develop resistance after an average year of treatment. There are several mechanisms for gefitinib and erlotinib to develop acquired resistance, including the secondary threonine 790 to methionine 790 mutation (T790M), also known as the "goalkeeper" T790M mutation (Xu y et al, cancer Biol ther.2010,9 (8): 572-582). Thus, second generation EGFR-TKI afatinib (afatinib) and third generation EGFR-TKI octtinib (osimertinib) (AZD 9291) were developed as irreversible EGFR inhibitors that bind to Cys797 for the treatment of T790M mutant patients. In particular, in NSCLC patients with EGFR T790M, substantially no WT EGFR-targeted octenib has achieved a higher clinical response rate. However, several recent studies reported tertiary Cys797 to Ser797 (C797S) point mutations accompanying the clinical treatment of octenib (Thress KS et al, nat. Med.2015,21 (6): 560-562.). There is a need for agents that overcome EGFR (C797S) resistance disorders in non-small cell lung cancer (NSCLC). EGFR targeting PROTAC is used as a potential strategy to overcome resistance mediated by these mutants, which has been disclosed or discussed in patent publications such as WO2018119441、WO2019149922、WO2019183523、WO2019121562、US20190106417、WO202157882、WO2021123087、WO2021133809、WO2021168074、WO2021208918 and WO 2021216440.

Nevertheless, a number of EGFR targets PROTAC designed for the degradation of EGFR muteins have been disclosed (Zhang X. Et al, eur.J.Med. Chem.2020,192, 112199; zhang H et al, eur.J.Med. Chem.2020,189, 112061; lu X, med. Res. Rev.2018,38 (5): 1550-1581.He K. Et al, biorg. Med. Chem. Lett.2020,15,127167.). Most of the molecules published are based on first, second and third generation EGFR inhibitors (WO 2021023233, WO2019121562 and WO 2018119441) or allosteric EGFR inhibitors (WO 2021127561). However, no data indicate that those EGFR targets PROTAC degrade all major EGFR mutations, such as Del19, L858R, del19/T790M, L858R/T790M, del/T790M/C797S, L858R/T790M/C797S.

The present application provides novel intermediates and methods of bifunctional compounds useful in the treatment of severe disease.

Disclosure of Invention

It is an object of the present invention to provide compounds and derivatives formed by conjugation of an EGFR inhibitor moiety with an E3 ligase ligand moiety, the compounds and derivatives functioning to recruit a target protein to the E3 ubiquitin ligase for degradation, and methods of making and using the same.

The compounds described herein or salts thereof are useful in the treatment of diseases that may be affected by EGFR modulation. The present application provides the use of a compound as described herein, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disease that may be affected by EGFR modulation. The application further provides a compound described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of a disease that may be affected by EGFR modulation. The application further provides a method of treating a proliferative disorder comprising administering to a subject in need thereof a therapeutically effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof.

In one embodiment, the difunctional compound is of formula (X):

or an N-oxide thereof, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a tautomer thereof, or a deuterated analog thereof, or a prodrug thereof,

Wherein:

E ¹ is N or CR ⁵;

e ² is N or CR ⁶;

R ^1a、R^1b、R^2a and R ^2b are each independently absent, hydrogen, halogen, -C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, -C _1-8 alkoxy, -C _3-8 cycloalkyl or-CN, each of said-C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, -C _1-8 alkoxy or-C _3-8 cycloalkyl being optionally substituted with at least one substituent selected from hydrogen, halogen, -C _1-8 alkoxy, -C _3-8 cycloalkyl or-CN;

R ³ and R ⁴ are each independently hydrogen, -C _1-6 alkyl or-C _3-8 cycloalkyl, each of said-C _1-6 alkyl or-C _3-8 cycloalkyl being optionally substituted with at least one substituent selected from hydrogen, halogen, -C _1-6 alkoxy;

R ⁵ and R ⁶ are each independently absent, hydrogen, halogen, -C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, -C _1-8 alkoxy, -C _3-8 cycloalkyl or-CN, each of said-C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, -C _1-8 alkoxy or-C _3-8 cycloalkyl being optionally substituted by at least one substituent selected from hydrogen, halogen, -C _1-8 alkoxy, -C _3-8 cycloalkyl or-CN, or

R ⁵ and R ⁶ together with the carbon atom to which they are attached form a 3 to 12 membered ring containing 0 to 3 heteroatoms independently selected from nitrogen, oxygen or sulfur, said ring optionally being substituted with at least one substituent halogen, hydroxy or-C ₁-C₈ alkyl;

R ⁷ is each independently absent, hydrogen, halogen, -C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, -C _1-8 alkoxy, -C _3-8 cycloalkyl or-CN, each of said-C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, -C _1-8 alkoxy or-C _3-8 cycloalkyl being optionally substituted by at least one substituent selected from hydrogen, halogen, -C _1-8 alkoxy, -C _3-8 cycloalkyl or-CN, or

Two R ⁷ taken together with the carbon atom to which they are attached form a 3-to 12-membered ring containing 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, said ring optionally substituted with at least one substituent halogen, hydroxy, or-C ₁-C₈ alkyl;

R ⁸ and R ⁹ are each independently selected from hydrogen, halogen, -C ₁-C₆ alkyl or C ₃-C₈ cycloalkyl, -C ₁-C₆ alkyl or C ₃-C₈ cycloalkyl each optionally substituted with at least one substituent selected from hydrogen, halogen, hydroxy, -C _1-6 alkoxy;

R ¹⁰ is each independently selected from hydrogen, halogen, -C ₁-C₈ alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, C ₆-C₁₂ aryl, 5 to 12 membered heteroaryl 、-NR^10aR^10b、-OR^10a、-SR^10a、-C(O)R^10a、-CO₂R^10a、-C(O)NR^10aR^10b、-NR^10aCOR^10b、-NR^10aCO₂R^10b or-NR ^10aSO₂R^10b or-CN, -C ₁-C₈ alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, C ₆-C₁₂ aryl or 5 to 12 membered heteroaryl each optionally substituted with at least one R ^10c;

R ^10a and R ^10b are each independently selected from hydrogen, -C ₁-C₈ alkyl, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, C ₆-C₁₂ aryl, or 5 to 12 membered heteroaryl, each of said-C ₁-C₈ alkyl, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, C ₆-C₁₂ aryl, or 5 to 12 membered heteroaryl optionally substituted with at least one substituent R ^10d;

R ^10c and R ^10d are each independently selected from halogen, hydrogen, -C ₁-C₈ alkyl, -C ₁-C₈ alkoxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, C ₆-C₁₂ aryl, 5 to 12 membered heteroaryl, oxo (＝O)、-NR^10eR^10f、-OR^10e、-SR^10e、-SO₂R^10e、-SO₂NR^10eR^10f、-C(O)R^10e、-CO₂R^10e、-C(O)NR^10eR^10f、-NR^10eCOR^10f、-NR^10eCO₂R^8f or-NR ^10eSO₂R^10f or-CN;

R ^10e and R ^10f are each independently selected from hydrogen, -C ₁-C₈ alkyl, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, C ₆-C₁₂ aryl, or 5 to 12 membered heteroaryl;

R ^11a、R^11b、R^11c、R^11d、R^12a、R^12b、R^12c and R ^12d are each independently absent, oxo, hydrogen, halogen, -C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, -C _1-8 alkoxy, or-C _3-8 cycloalkyl, -C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, -C _1-8 alkoxy, or-C _3-8 cycloalkyl, each optionally substituted with at least one substituent selected from hydrogen, halogen, -C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, -C _1-8 alkoxy, or-CN;

L ¹ is independently selected from -O-、-NR^a-、-C(O)-、*^L1-C(O)NR^a-**^L1、*^L1-C(O)O-**^L1、*^L1-NR^aC(O)-**^L1、*^L1-OC(O)-**^L1、

Wherein said

Each optionally substituted with at least one R ^L1c;

wherein ^L1 is the sum of The position of the partial connection, and ^L1 refers to the position of the connection withThe position of the partial connection;

L ² is independently selected from -O-、-NR^a-、-C(O)-、*^L2-C(O)NR^a-**^L2、*^L2-C(O)O-**^L2、*^L2-NR^aC(O)-**^L2、*^L2-OC(O)-**^L2、

Wherein said Each optionally substituted with at least one R ^L2c;

Wherein ^L2 is the sum of The position of the partial connection, and ^L2 refers to the position of the connection withThe position of the partial connection;

l ³ is independently selected from -O-、-NR^a-、-C(O)-、*^L3-C(O)NR^a-**^L3、*^L3-C(O)O-**^L3、*^L3-NR^aC(O)-**^L3、*^L3-OC(O)-**^L3、 Wherein said Each optionally substituted with at least one R ^L3c;

wherein ^L3 is the sum of The position of the partial connection, and ^L3 refers to the position of the connection withThe position of the partial connection;

The R ^L1c、R^L2c and R ^L3c are each independently absent, oxo (= O), halogen, hydroxy, -CN, -C ₁-C₈ alkyl, -C ₁-C₈ alkoxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, C ₃-C₈ cycloalkyl, 3-to 8-membered heterocyclyl, C ₆-C₁₂ aryl or 5-to 12-membered heteroaryl, the-C ₁-C₈ alkyl, -C ₁-C₈ alkoxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, C ₃-C₈ cycloalkyl, 3-to 8-membered heterocyclyl, C ₆-C₁₂ aryl and 5-to 12-membered heteroaryl are each optionally substituted by at least one R ^Lca,

R ^Lca is independently absent, oxo (=O), halogen, hydroxy, -CN, -C ₁-C₈ alkyl, -C ₁-C₈ alkoxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, C ₃-C₈ cycloalkyl, 3-to 8-membered heterocyclyl, C ₆-C₁₂ aryl, or 5-to 12-membered heteroaryl, or

Two R ^L1c taken together with the atoms to which they are attached form a 3-to 12-membered ring containing 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, said ring optionally substituted with at least one substituent halogen, hydroxy, -C ₁-C₈ alkyl;

Two R ^L2c taken together with the atoms to which they are attached form a 3-to 12-membered ring containing 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, said ring optionally substituted with at least one substituent halogen, hydroxy, -C ₁-C₈ alkyl;

two R ^L3c taken together with the atoms to which they are attached form a 3-to 12-membered ring containing 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, said ring optionally substituted with at least one substituent halogen, hydroxy, -C ₁-C₈ alkyl;

Selected from the group consisting of

Z ¹,Z² and Z ³ are each independently N or CR ^z, provided that Z ¹、Z² and Z ³ are not both N;

R ^z is independently selected at each occurrence from the group consisting of absent, hydrogen, halogen, -C _1-8 alkyl, -NR ^ZaR^Zb、-OR^Za、-SR^Za、C₃-C₈ cycloalkyl, 3-to 8-membered heterocyclyl, or CN, -C _1-8 alkyl, C ₃-C₈ cycloalkyl, 3-to 8-membered heterocyclyl each optionally substituted with at least one R ^Zc;

Part is linked to either Z ¹ or Z ² (which is CR ^z and R ^z is absent) A portion;

R ^Za and R ^Zb are each independently selected from the group consisting of absent, hydrogen, -C ₁-C₈ alkyl, C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, C ₆-C₁₂ aryl, or 5 to 12 membered heteroaryl, each of said-C _1-8 alkyl, C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, C ₆-C₁₂ aryl, or 5 to 12 membered heteroaryl optionally substituted with at least one substituent R ^Zd;

R ^Zc and R ^Zd are each independently halogen, hydroxy, -C ₁-C₈ alkyl, -C _1-8 alkoxy, C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, C ₆-C₁₂ aryl, or 5 to 12 membered heteroaryl;

R ¹³ and R ¹⁴ are each independently selected from the group consisting of absent, hydrogen, halogen, -C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, -C _1-8 alkoxy, -C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, -C ₆-C₁₂ aryl, 5 to 12 membered heteroaryl 、-CN、-SO₂R^13a、-SO₂NR^13aR^13b、-COR^13a,-CO₂R^13a、-CONR^13aR^13b、-NR^13aR^13b、-NR^13aCOR^13b、-NR^13aCO₂R^13b or-NR ^13aSO₂R^13b;-C_1-8 alkyl-C _2-8 alkenyl, -C _2-8 alkynyl, -C _1-8 alkoxy, -C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, -C ₆-C₁₂ aryl or 5 to 12 membered heteroaryl each optionally substituted with halogen, -C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, -C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, C ₆-C₁₂ aryl, 5 to 12 membered heteroaryl, oxo 、-CN、-OR^13c、-SO₂R^13c、-SO₂NR^13cR^13d、-COR^13c、-CO₂R^13c、-CONR^13cR^13d、-NR^13cR^13d、-NR^13cCOR^13d、-NR^13cCO₂R^13d or-NR ^13cSO₂R^13d;

Each occurrence of R ^13a、R^13b、R^13c and R ^13d is independently absent, hydrogen, -C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, C ₆-C₁₂ aryl, or 5 to 12 membered heteroaryl;

L ⁴、L⁵ and L ⁶ are each independently selected from the group consisting of absent, single bond 、-O-、-NR^a-、-(CR^aR^b)_n8-、-O(CR^aR^b)_n8-、-NR^a(CR^aR^b)_n8-, or-C (O) -;

At each occurrence, X ¹、X² and X ⁷ are each independently selected from-CR ^a or N;

At the time of each occurrence of this, X ³、X⁴ and X ⁸ are each independently selected from the group consisting of-NR ^a -; -O-, -S-and-CR ^aR^b -;

At each occurrence, X ⁵ and X ⁶ are each independently selected from the group consisting of absent, single bond, -C (O) -, -NR ^a -, and-O-;

At each occurrence, R ^a and R ^b are each independently selected from hydrogen, hydroxy, halogen, CN, -C ₁-C₈ alkyl, -C ₁-C₈ alkoxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, -C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, -C ₆-C₁₂ aryl or 5 to 12 membered heteroaryl, said-C ₁-C₈ alkyl, -C ₁-C₈ alkoxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, -C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, -C ₆-C₁₂ aryl or 5 to 12 membered heteroaryl each optionally substituted with at least one substituent halogen, hydroxy, halogen, -C ₁-C₈ alkyl, -C ₁-C₈ alkoxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, -C ₃-C₈ cycloalkyl, -3 to 8 membered heterocyclyl, -C ₆-C₁₂ aryl or 5 to 12 membered heteroaryl substitution, or

R ^a and R ^b together with the carbon atom to which they are attached form a3 to 12 membered ring containing 0-3 heteroatoms independently selected from nitrogen, oxygen or sulfur, said ring optionally being substituted with at least one substituent halogen, hydroxy, -C ₁-C₈ alkyl, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, -C ₁-C₈ alkoxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, C ₃-C₈ cycloalkyl, 3 to 8 membered heterocyclyl, C ₆-C₁₂ aryl or 5 to 12 membered heteroaryl;

m1, m2, m3 and m4 are each independently 0, 1 or 2, provided that m1+m2+m3+m4.ltoreq.4;

m5, m6 and m7 are each independently 0, 1 or 2, provided that m5+m6+m7.gtoreq.1;

n1, n2, n3, n4 and n5 are each independently 0, 1,2 or 3;

n6 are each independently 0,1, 2, 3 or 4;

n7 and n8 are each independently 0, 1, 2 or 3;

s1 and s2 are each independently 0, 1, 2 or 3;

s5, s6 and s7 are each independently 0, 1,2 or 3;

Provided that it is

For either L ¹、L² or L ³, X ⁵ is a single bond, absent, -C (O) -, when X ¹ is N, and/or X ⁶ is a single bond, absent, -C (O) -, when X ² is N;

When L ¹ is When X ¹ is N, X ⁵ is a single bond, absent, -C (O) -, and/or X ³ is-CR ^aR^b -, when X ² is N, X ⁶ is a single bond, absent, -C (O) -, and/or X ⁴ is-CR ^aR^b -;

When L ² is When X ¹ is N, X ⁵ is a single bond, absent, -C (O) -, and/or X ³ is-CR ^aR^b -, when X ² is N, X ⁶ is a single bond, absent, -C (O) -, and/or X ⁴ is-CR ^aR^b -;

When L ³ is When X ¹ is N, X ⁵ is a single bond, absent, -C (O) -, and/or X ³ is-CR ^aR^b -, when X ² is N, X ⁶ is a single bond, absent, -C (O) -, and/or X ⁴ is-CR ^aR^b -.

It is another object of the present invention to provide intermediates and methods of compound (X) formed by conjugation of an EGFR inhibitor moiety with an E3 ligase ligand moiety, the function of which is to recruit a target protein to the E3 ubiquitin ligase for degradation, and methods of making and using the same.

Aspect 1. A compound of formula (I):

Wherein:

R ⁸ and R ⁹ are each independently selected from hydrogen, halogen, -C ₁-C₆ alkyl or C ₃-C₈ cycloalkyl, -C ₁-C₆ alkyl or C ₃-C₈ cycloalkyl each optionally substituted with at least one substituent selected from hydrogen, halogen, -C _1-6 alkoxy;

r ¹⁰ are each independently selected from halogen, Wherein said

Each optionally substituted with at least one R ^10a;

Each of said R ^10a is independently absent, oxo (= O), halogen, hydroxy, -CN, -C ₁-C₈ alkyl, -C ₁-C₈ alkoxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, C ₃-C₈ cycloalkyl, 3-to 8-membered heterocyclyl, C ₆-C₁₂ aryl or 5-to 12-membered heteroaryl, said-C ₁-C₈ alkyl, -C ₁-C₈ alkoxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, C ₃-C₈ cycloalkyl, 3-to 8-membered heterocyclyl, C ₆-C₁₂ aryl and 5-to 12-membered heteroaryl each optionally substituted with at least one R ^10b,

R ^10b is independently absent, oxo (=O), halogen, hydroxy, -CN, -C ₁-C₈ alkyl, -C ₁-C₈ alkoxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, C ₃-C₈ cycloalkyl, 3-to 8-membered heterocyclyl, C ₆-C₁₂ aryl, or 5-to 12-membered heteroaryl, or

Two R ^10a taken together with the atoms to which they are attached form a 3-to 12-membered ring containing 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, said ring optionally substituted with at least one substituent halogen, hydroxy, -C ₁-C₈ alkyl;

Each occurrence of X ¹ is independently selected from-CR ^a or N;

Each occurrence of X ² is independently selected from the group consisting of-NH-or-C (O) -;

At the time of each occurrence of this, X ³ are each independently selected from the group consisting of-NR ^a -; -O-, -S-and-CR ^aR^b -;

n1, n2, n3, n4 and n5 are each independently 0, 1,2 or 3;

s5, s6 and s7 are each independently 0, 1,2 or 3.

Aspect 2 the compound according to aspect 1, wherein the compound is selected from the group consisting of formula (IIa),

Preferably, the compound is selected from formulas (IIb) and (IIc):

Preferably, the compound is selected from formulas (IId) and (IIe):

preferably, the compound is selected from formulas (IIf) and (Ig):

Wherein R ³、R⁴、R⁷、R⁸、R⁹、R¹⁰, s5, s6 and s7 are as defined in aspect 1.

The compound of any one of the preceding aspects, wherein each of R ^1a、R^1b、R^2a and R ^2b is independently hydrogen, F, cl, br, I, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, -C _2-8 alkenyl, -C _2-8 alkynyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, hexyloxy, heptyloxy, octyloxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, or-CN, wherein each of the methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, -C _2-8 alkenyl, -C _2-8 alkynyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, hexyloxy, heptyloxy, octyloxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl is optionally substituted with at least one substituent selected from hydrogen, F, cl, br, I, methoxy, ethoxy, propoxy, butoxy, pentoxy, hexyloxy, heptyloxy, octyloxy, cyclopropyl, cyclohexyl, cyclooctyl;

Preferably, R ^1a、R^1b、R^2a and R ^2b are each independently hydrogen, F, cl, br, I, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, hexoxy, heptoxy, octoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl 、-CF₃、-CHF₂、-CN、-CH₂OCH₃、-CH₂OCH₂CH₃、-CH₂CH₂OCH₃、-CH₂CH₂OCH₂CH₃;

More preferably, R ^1a、R^1b、R^2a and R ^2b are each independently hydrogen, F, cl, methyl, methoxy, cyclopropyl, -CF ₃, or-CHF ₂、-CH₂OCH₃;

More preferably, R ^1a、R^1b、R^2a and R ^2b are each independently hydrogen.

A compound according to any one of the preceding aspects wherein R ³ and R ⁴ are each independently hydrogen, methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl, the methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl being optionally substituted with at least one substituent selected from hydrogen, F, cl, br, I, methoxy, ethoxy, propoxy, butoxy, pentoxy or hexoxy;

Preferably, R ³ and R ⁴ are each independently hydrogen, methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl, preferably R ³ is independently methyl and R ⁴ is hydrogen.

Aspect 5. The compound of any one of the preceding aspects, wherein each R ⁷ is independently hydrogen, F, cl, br, I, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, hexoxy, heptoxy, octoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl 、-CF₃、-CHF₂、-CN、-CH₂OCH₃、-CH₂OCH₂CH₃、-CH₂CH₂OCH₃、-CH₂CH₂OCH₂CH₃;

Preferably, each R ⁷ is independently hydrogen, F, cl, methyl, methoxy, cyclopropyl, -CF ₃, or-CHF ₂、-CH₂OCH₃.

Aspect 6. The compound of any of the preceding aspects, wherein two R ⁷ form, with the carbon atom to which they are attached, a 3, 4, 5, 6, 7, or 8 membered ring containing 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, the ring optionally being substituted with at least one substituent F, cl, br, I, hydroxy, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, or octyl;

Preferably, two R ⁷ form a3, 4, 5, or 6 membered ring with the carbon atom to which they are attached, said ring being optionally substituted with at least one substituent F, cl, br, I, hydroxy, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, or octyl.

Aspect 7. The compound of any one of the preceding aspects, wherein each of R ⁸ and R ⁹ is independently selected from hydrogen, F, cl, br, I, methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl, the methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl being optionally substituted with at least one substituent selected from hydrogen, F, cl, br, I, methoxy, ethoxy, propoxy, butoxy, pentoxy, or hexoxy;

Preferably, R ⁸ and R ⁹ are each independently hydrogen, F, cl, br, I, methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl, preferably, R ⁸ is independently hydrogen and R ⁹ is F or methyl.

Aspect 8 the compound of any one of the preceding aspects, wherein each R ¹⁰ is independently selected from hydrogen, -F, -Cl, -Br, -I,

Wherein said

Each optionally substituted with at least one R ^10a;

Each R ^10a is independently hydrogen, oxo (= O), F, cl, br, I, hydroxy, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, hexyloxy, heptyloxy, octyloxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 3 to 8 membered heterocyclyl, phenyl or 5 to 12 membered heteroaryl, each of said methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, hexyloxy, heptyloxy, octyloxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 3 to 8 membered heterocyclyl, phenyl and 5 to 12 membered heteroaryl being optionally substituted by at least one R ^10b,

R ^10b is independently oxo (= O), F, cl, br, I, hydroxy, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, hexyloxy, heptyloxy, octyloxy, -C ₂-C₈ alkenyl, -C ₂-C₈ alkynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 3-to 8-membered heterocyclyl, phenyl or 5-to 12-membered heteroaryl, or

Two R ^10a taken together with the carbon atom to which they are attached form a 3, 4, 5, 6, 7, or 8 membered ring containing 0, 1, 2, or 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, said ring optionally substituted with at least one substituent F, cl, br, I, hydroxy, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, or octyl;

Preferably, R ¹⁰ is each independently selected from hydrogen, -F, -Cl, -Br, -I, I,

Aspect 9A compound according to any one of the preceding aspects selected from

Detailed Description

The following terms have the indicated meanings throughout the specification:

unless defined otherwise herein, all other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

The following terms have the indicated meanings throughout the specification:

As used herein, including the appended claims, the singular forms of words such as "a," "an," and "the" include their corresponding plural referents unless the context clearly dictates otherwise.

The term "or" is used to mean, and is used interchangeably with, the term "and/or" unless the context clearly dictates otherwise.

The term "alkyl" includes hydrocarbon groups selected from straight and branched chain saturated hydrocarbon groups containing from 1 to 18 (such as from 1 to 12, further such as from 1 to 10, still further such as from 1 to 8, or from 1 to 6, or from 1 to 4) carbon atoms. Examples of alkyl groups containing 1 to 6 carbon atoms (i.e., C _1-6 alkyl groups) include, but are not limited to, methyl, ethyl, 1-propyl or n-propyl ("n-Pr"), 2-propyl or isopropyl ("i-Pr"), 1-butyl or n-butyl ("n-Bu"), 2-methyl-1-propyl or isobutyl ("i-Bu"), 1-methylpropyl or sec-butyl ("s-Bu"), 1-dimethylethyl or tert-butyl ("t-Bu"), 1-pentyl, 2-pentyl, 3-pentyl, 2-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, 3-dimethyl-2-butyl, and 3, 3-dimethyl-2-butyl.

The term "propyl" includes 1-propyl or n-propyl ("n-Pr"), 2-propyl or isopropyl ("i-Pr").

The term "butyl" includes 1-butyl or n-butyl ("n-Bu"), 2-methyl-1-propyl or isobutyl ("i-Bu"), 1-methylpropyl or sec-butyl ("s-Bu"), 1-dimethylethyl or tert-butyl ("t-Bu").

The term "pentyl" includes 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl.

The term "hexyl" includes 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2, 3-dimethyl-2-butyl and 3, 3-dimethyl-2-butyl.

The term "alkylene" refers to a divalent alkyl group produced by removing two hydrogens from an alkane. Alkylene groups include, but are not limited to, methylene, ethylene, propylene, and the like.

The term "halogen" includes fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

The term "alkenyl" includes hydrocarbyl groups selected from straight and branched chain hydrocarbyl groups containing at least one c=c double bond and from 2 to 18 (such as from 2 to 8, further such as from 2 to 6) carbon atoms. Examples of alkenyl groups (e.g., C _2-6 alkenyl groups) include, but are not limited to, vinyl (ethyl or vinyl), prop-1-enyl, prop-2-enyl, 2-methylprop-1-enyl, but-2-enyl, but-3-enyl, but-1, 3-dienyl, 2-methylbut-1, 3-dienyl, hex-1-enyl, hex-2-enyl, hex-3-enyl, hex-4-enyl, and hex-1, 3-dienyl.

The term "alkenylene" refers to a divalent alkenyl group produced by removing two hydrogens from an alkene. Alkenylene includes, but is not limited to, vinylidene, butenylene, and the like.

The term "alkynyl" includes hydrocarbyl groups selected from straight and branched chain hydrocarbyl groups containing at least one c≡c triple bond and from 2 to 18 (such as from 2 to 8, further such as from 2 to 6) carbon atoms. Examples of alkynyl groups (e.g., C _2-6 alkynyl) include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl (propargyl), 1-butynyl, 2-butynyl, and 3-butynyl.

The term "alkynylene" refers to a divalent alkynyl group produced by removing two hydrogens from an alkyne. Alkynylene includes, but is not limited to, ethynylene and the like.

The term "cycloalkyl" includes hydrocarbyl groups selected from saturated cyclic hydrocarbyl groups, including monocyclic and polycyclic (e.g., bicyclic and tricyclic) groups, including fused, bridged, or spiro cycloalkyl groups.

For example, cycloalkyl groups may contain 3 to 12 (such as 3 to 10, further such as 3 to 8, further such as 3 to 6, 3 to 5, or 3 to 4) carbon atoms. Even further for example, cycloalkyl groups may be selected from monocyclic groups containing 3 to 12 (such as 3 to 10, further such as 3 to 8, 3 to 6) carbon atoms. Examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl and cyclododecyl. In particular, examples of saturated monocyclic cycloalkyl groups (e.g., C _3-8 cycloalkyl groups) include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. In a preferred embodiment, cycloalkyl is a monocyclic ring containing 3 to 6 carbon atoms (abbreviated to C _3-6 cycloalkyl), including, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. Examples of bicyclic cycloalkyl groups include those having 7 to 12 ring atoms arranged as a fused bicyclic ring selected from the group consisting of [4,4], [4,5], [5,6] and [6,6] ring systems, or as a bridged bicyclic ring selected from the group consisting of bicyclo [2.2.1] heptane, bicyclo [2.2.2] octane and bicyclo [3.2.2] nonane. Other examples of bicyclic cycloalkyl groups include those arranged as bicyclic rings selected from the group consisting of the [5,6] and [6,6] ring systems.

The term "spirocycloalkyl" includes cyclic structures containing carbon atoms and formed by at least two rings sharing one atom.

The term "fused cycloalkyl" includes bicyclic cycloalkyl groups as defined herein which are saturated and formed by two or more rings sharing two adjacent atoms.

The term "bridged cycloalkyl" includes cyclic structures containing carbon atoms and formed by two rings that share two atoms that are not adjacent to each other. The term "7-to 10-membered bridged cycloalkyl" includes cyclic structures containing 7 to 12 carbon atoms and formed by two rings sharing two atoms that are not adjacent to each other.

Examples of fused cycloalkyl, fused cycloalkenyl, or fused cycloalkynyl include, but are not limited to, bicyclo [1.1.0] butyl, bicyclo [2.1.0] pentyl, bicyclo [3.1.0] hexyl, bicyclo [4.1.0] heptyl, bicyclo [3.3.0] octyl, bicyclo [4.2.0] octyl, decalin, and benzo3 to 8 membered cycloalkyl, benzoC _4-6 cycloalkenyl, 2, 3-dihydro-1H-indenyl, 1,2,3, 4-tetrahydronaphthyl, 1, 4-dihydronaphthyl, and the like. Preferred embodiments are 8 to 9 membered fused rings, which in the above examples refer to cyclic structures containing 8 to 9 ring atoms.

The term "aryl" used alone or in combination with other terms includes a group selected from the group consisting of:

5-and 6-membered carbocyclic aromatic rings, for example, phenyl;

bicyclic ring systems, such as 7-to 12-membered bicyclic ring systems, wherein at least one ring is carbocyclic and aromatic, e.g., naphthyl and indanyl, and

Tricyclic ring systems, such as 10 to 15 membered tricyclic ring systems, wherein at least one ring is carbocyclic and aromatic, e.g., fluorenyl.

The terms "aromatic hydrocarbon ring" and "aryl" are used interchangeably throughout the disclosure herein. In some embodiments, the monocyclic or bicyclic aromatic hydrocarbon ring has 5 to 10 ring-forming carbon atoms (i.e., a C _5-10 aryl group). Examples of monocyclic or bicyclic aromatic hydrocarbon rings include, but are not limited to, phenyl, naphthalen-1-yl, naphthalen-2-yl, anthracenyl, phenanthrenyl, and the like. In some embodiments, the aromatic hydrocarbon ring is a naphthalene ring (naphthalen-1-yl or naphthalen-2-yl) or a phenyl ring. In some embodiments, the aromatic hydrocarbon ring is a phenyl ring.

In particular, the term "bicyclic fused aryl" includes bicyclic aryl rings as defined herein. A typical bicyclic fused aryl is naphthalene.

The term "heteroaryl" includes groups selected from the group consisting of:

A 5-, 6-, or 7-membered aromatic monocyclic ring comprising at least one heteroatom selected from nitrogen (N), sulfur (S), and oxygen (O), e.g., 1 to 4 (or in some embodiments, 1 to 3, in some embodiments, 1 to 2) heteroatoms, the remaining ring atoms being carbon;

A 7-to 12-membered bicyclic ring comprising at least one heteroatom selected from N, O and S, for example 1 to 4 (or in some embodiments 1 to 3, or in other embodiments 1 or 2) heteroatoms, the remaining ring atoms being carbon, and wherein at least one ring is aromatic and at least one heteroatom is present in an aromatic ring, and

11-To 14-membered tricyclic ring containing at least one heteroatom selected from N, O and S, such as 1 to 4 (or in some embodiments 1 to 3, or in other embodiments 1 or 2) heteroatoms, the remaining ring atoms being carbon, and wherein at least one ring is aromatic and at least one heteroatom is present in an aromatic ring.

When the total number of S and O atoms in the heteroaryl group exceeds 1, those heteroatoms are not adjacent to each other. In some embodiments, the total number of S and O atoms in the heteroaryl group is no more than 2. In some embodiments, the total number of S and O atoms in the aromatic heterocycle is no more than 1. When the heteroaryl group contains more than one heteroatom ring member, the heteroatoms may be the same or different. The nitrogen atoms in one or more rings of the heteroaryl group may be oxidized to form an N-oxide.

In particular, the term "bicyclic fused heteroaryl" includes 7 to 12 membered, preferably 7 to 10 membered, more preferably 9 or 10 membered fused bicyclic heteroaryl rings as defined herein. Typically, bicyclic fused heteroaryl groups are 5-membered/5-membered, 5-membered/6-membered, 6-membered/6-membered or 6-membered/7-membered bicyclic. The group may be attached to the remainder of the molecule through any ring.

"Heterocyclyl", "heterocycle" or "heterocyclic" are interchangeable and include non-aromatic heterocyclyl groups containing one or more heteroatoms selected from nitrogen, oxygen or optionally oxidized sulfur as ring members, the remaining ring members being carbon, including monocyclic, fused, bridged and spiro rings, i.e., containing monocyclic, bridged, spiro and fused heterocyclyl groups.

The term "H" or "hydrogen" as disclosed herein includes hydrogen and the nonradioactive isotope deuterium.

The term "at least one substituent" as disclosed herein includes, for example, 1 to 4 (such as 1 to 3, further such as 1 or 2) substituents, provided that valence theory is met. For example, the "at least one substituent F" disclosed herein includes 1 to 4 (such as 1 to 3, further such as 1 or 2) substituents F.

The term "divalent" refers to a linking group capable of forming a covalent bond with two other moieties. For example, "divalent cycloalkyl" refers to cycloalkyl obtained by removing two hydrogens from the corresponding cycloalkane to form a linking group. The terms "divalent aryl", "divalent heterocyclic" or "divalent heteroaryl" are to be understood in a similar manner.

The compounds disclosed herein may contain asymmetric centers and thus may exist as enantiomers. "enantiomer" refers to two stereoisomers of a compound that are mirror images of each other that are not stackable. Where the compounds disclosed herein have two or more asymmetric centers, they may additionally exist as diastereomers. Enantiomers and diastereomers belong to a broader class of stereoisomers. It is intended to include all such possible stereoisomers in the form of substantially pure resolved enantiomers, racemic mixtures thereof and mixtures of diastereomers. It is intended to include all stereoisomers of the compounds disclosed herein and/or pharmaceutically acceptable salts thereof. Unless otherwise specifically indicated, references to one isomer apply to any possible isomer. All possible isomers are included as long as the isomer composition is not specified.

Unless otherwise indicated, when a compound disclosed herein contains an olefinic double bond, such double bond is intended to include both E and Z geometric isomers.

When the compounds disclosed herein contain disubstituted cyclic ring systems, the substituents found on such ring systems may take both cis and trans forms. The cis form means that both substituents are located on the upper side of the 2 substituent positions on the carbon, while the trans form means that they are located on opposite sides. For example, the disubstituted cyclic ring system may be a cyclohexyl ring or a cyclobutyl ring.

It may be advantageous to separate the reaction products from each other and/or from the starting materials. The desired product of each step or series of steps is isolated and/or purified (hereinafter isolated) to the desired degree of homogeneity by techniques commonly used in the art. Typically, such separations involve multiphase extraction, crystallization from a solvent or solvent mixture, distillation, sublimation, or chromatography. Chromatography may involve any number of methods including, for example, reversed and normal phase chromatography, size exclusion chromatography, ion exchange chromatography, high, medium and low pressure liquid chromatography methods and apparatus, small scale analytical chromatography, simulated moving bed ("SMB") chromatography and preparative thin or thick layer chromatography, as well as small scale thin and flash chromatography techniques. Those skilled in the art may select and apply the techniques most likely to achieve the desired separation.

"Diastereoisomers" refers to stereoisomers of a compound having two or more chiral centers that are not mirror images of each other. Mixtures of diastereomers can be separated into their individual diastereomers based on their physicochemical differences by methods well known to those skilled in the art, such as by chromatography and/or fractional crystallization. Enantiomers may be isolated by reaction with an appropriate optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher's acid chloride) to convert the enantiomeric mixture to a diastereomeric mixture, separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereomers to the corresponding pure enantiomers. Enantiomers can also be separated by using chiral HPLC columns.

Single stereoisomers (e.g., substantially pure enantiomers) may be obtained by resolution of a racemic mixture using a method such as the formation of diastereomers using optically active resolving agents (Eliel, E. And Wilen, S.Stereochemistry of Organic Compounds. New York: john Wiley & Sons, inc.,1994; lochmuller, C.H. et al, "Chromatographic resolution of enantiomers: selective review," J.color., 113 (3) (1975): pages 283-302). The racemic mixture of the chiral compounds of the present invention can be separated and isolated by any suitable method including (1) formation of ionic diastereomeric salts with the chiral compounds and separation by fractional crystallization or other means, (2) formation of diastereomeric compounds with chiral derivatizing reagents, separation of diastereomers and conversion to pure stereoisomers, and (3) separation of the substantially pure or enriched stereoisomers directly under chiral conditions. See Wainer, irving W.code, drug Stereochemistry: ANALYTICAL METHODS AND Pharmacology New York: MARCEL DEKKER, inc.,1993.

Some of the compounds disclosed herein may exist with different hydrogen attachment points, known as tautomers. For example, compounds comprising a carbonyl-CH ₂ C (O) -group (keto form) may undergo tautomerism to form a hydroxy-ch=c (OH) -group (enol form). Where applicable, it is also intended to include both the individual keto and enol forms as well as mixtures thereof. As another example, compounds comprising pyrazolyl groups may undergo tautomerism to form different rings, as shown below:

as another example, compounds that include a guanidino group in the ring may undergo tautomerism to form a different ring, as shown below:

"prodrug" refers to an active agent derivative that requires conversion in vivo to release the active agent. In some embodiments, the transformation is enzymatic. Prodrugs are typically (but not necessarily) pharmacologically inactive prior to conversion to the active agent.

"Deuterated analog" refers to a derivative of an active agent in which any hydrogen is replaced by deuterium. In some embodiments, the deuterated position is located on the warhead portion. In some embodiments, the deuterated position is located on a linker moiety. In some embodiments, the deuterated position is located on the down-resolution stator portion.

By "pharmaceutically acceptable salts" is meant those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts can be prepared in situ during the final isolation and purification of the compounds disclosed herein, or separately by reacting the free base functionality with a suitable organic acid or by reacting the acidic group with a suitable base. The term also includes salts of stereoisomers (such as enantiomers and/or diastereomers), tautomers and prodrugs of the compounds of the invention.

In addition, if the compounds disclosed herein are obtained as acid addition salts, the free base can be obtained by basifying a solution of the acid salt. Conversely, if the product is a free base, the addition salt, such as a pharmaceutically acceptable addition salt, can be prepared by dissolving the free base in a suitable organic solvent and treating the solution with an acid according to conventional methods for preparing acid addition salts from base compounds. Those of skill in the art will recognize a variety of synthetic methods that can be used to prepare non-toxic pharmaceutically acceptable addition salts without undue experimentation.

When applied to an animal, human, experimental subject, cell, tissue, organ, or biological fluid, the terms "administration", "Administration (ADMINISTERING)", "treatment" and "treatment" herein mean the contacting of an exogenous drug, therapeutic, diagnostic agent, or composition with the animal, human, subject, cell, tissue, organ, or biological fluid. Treatment of a cell encompasses contact of a reagent with a cell, as well as contact of a reagent with a fluid, wherein the fluid is in contact with the cell. The terms "administering" and "treatment" also mean the in vitro and ex vivo treatment of a cell, e.g., by an agent, diagnostic agent, binding compound, or by another cell. The term "subject" herein includes any organism, preferably an animal, more preferably a mammal (e.g., rat, mouse, dog, cat, and rabbit), and most preferably a human.

The term "treatment" or "treatment" as used herein also generally refers to obtaining a desired pharmacological and/or physiological effect. The effect may be prophylactic according to total or partial prophylaxis of the disease or symptoms thereof, and/or therapeutic according to partial or total stabilization or cure of the disease and/or side effects caused by the disease. As used herein, "treatment" or "treatment" encompasses any treatment of a disease in a patient, including (a) preventing a disease or condition in a patient who may be predisposed to the disease or condition but who has not yet been diagnosed with the disease or condition, (b) inhibiting the symptoms of the disease, i.e., preventing its progression, or (c) alleviating the symptoms of the disease, i.e., causing complete or partial regression of the disease or condition.

The term "effective amount" or "therapeutically effective amount" refers to an amount of an active ingredient (such as a compound) that, when administered to a subject to treat a disease, or at least one clinical symptom of a disease or disorder, is sufficient to affect treatment for the disease, disorder or symptom. The term "therapeutically effective amount" may vary with the compound, the disease, the disorder, and/or the symptoms of the disease or disorder, the disease, the disorder, and/or the severity of the symptoms of the disease or disorder, the age of the subject to be treated, and/or the weight of the subject to be treated. The appropriate amount may be readily apparent to one of ordinary skill in the art in any given situation, or may be determined by routine experimentation. In some embodiments, a "therapeutically effective amount" is an amount of at least one compound disclosed herein and/or at least one stereoisomer, tautomer, or prodrug thereof and/or at least one pharmaceutically acceptable salt thereof that is effective to "treat" a disease or disorder in a subject as defined herein. In the case of combination therapies, the term "therapeutically effective amount" refers to the total amount of the combination subject used to effectively treat the disease, disorder, or condition.

The term "disease" refers to any disease, disorder, illness, symptom, or indication, and is interchangeable with the term "disorder" or "condition.

Throughout the specification and the claims which follow, unless the context requires otherwise, the term "comprise" and variations such as "comprises" and "comprising" are intended to specify the presence of the following features but do not preclude the presence or addition of one or more other features. As used herein, the term "comprising" may be replaced with the terms "including", "comprising" or sometimes with "having".

Throughout the specification and the appended claims, the term "C _n-m" or "C _n-C_m" is indicative of a range including endpoints, where n and m are integers and indicates the number of carbons. Examples include C _1-8、C_1-6、C₁-C₈、C₁-C₆, and the like.

Percentages, ratios, or parts used in the present application are by weight or volume unless otherwise specified. The amount used in the present application is a weight or volume amount. Which can be readily determined by one skilled in the art.

Hereinafter, the present application will be described with reference to examples, which illustrate advantageous effects of the present application. Those skilled in the art will recognize that these embodiments are illustrative and not limiting. These examples are not intended to limit the scope of the application in any way. The experimental methods described in the examples below are conventional except as specified, and the reagents and materials used are commercially available except as specified.

Examples

The following examples are intended to be illustrative only and should not be construed as limiting in any way. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.), but some experimental errors and deviations should be accounted for. Unless indicated otherwise, temperatures are in degrees celsius. Reagents were purchased from commercial suppliers such as Sigma-Aldrich, ALFA AESAR or TCI and used without further purification unless indicated otherwise. Unless otherwise indicated, the reactions set forth below were carried out under positive pressure nitrogen or argon or with a desiccation tube in anhydrous solvent, the reaction flask was fitted with a rubber septum for introduction of the substrate and reagents via syringe, and glassware oven and/or heat dried.

¹ The H NMR spectra were recorded on an Agilent instrument operating at 400 MHz. ¹ HNMR (high-density magnetic resonance)

Spectra were obtained using CDCl ₃、CD₂Cl₂、CD₃OD、D₂O、d₆-DMSO、d₆ -acetone or (CD ₃)₂ CO as solvent and tetramethylsilane (0.00 ppm) or residual solvent (CDCl₃：7.25ppm;CD₃OD：3.31ppm;D₂O：4.79ppm;d₆-DMSO：2.50ppm;d₆- acetone: 2.05, (CD ₃)₃ CO: 2.05) when peak multiplets are reported, the abbreviations s (singlet), d (doublet), t (triplet), q (quartet), qn (quintet), sx (sextuply), m (multiplet), br (broad), dd (doublet), dt (doublet) when given are reported in hertz (Hz).

LCMS-1 LC-MS spectrometer (Agilent 1260 Informance), detector: MWD (190-400 nm), mass detector: 6120SQ, mobile phase: A: water with 0.1% formic acid, B: acetonitrile with 0.1% formic acid, column: poroshell EC-C18, 4.6X105 mm,2.7pm, gradient method: flow: 1.8mL/min, time (min) A (%) B (%)

LCMS, LCMS-3:LC-MS spectrometer (Agilent 1260 Informance II), detector: MWD (190-400 nm), mass detector: G6125C SQ, mobile phase: A: water containing 0.1% formic acid, B: acetonitrile containing 0.1% formic acid, column: poroshell EC-C18, 4.6X105 mm,2.7pm, gradient method: flow: 1.8mL/min, time (min) A (%) B (%)

Time (min)	A(%)	B(%)
			0.00	95	5
1.5	5	95
			2.0	5	95
2.1	95	5
			3.0	95	5

LCMS-2 LC-MS spectrometer (Agilent 1290 Infinicity II), detector: MWD (190-400 nm), mass detector: G6125 CSQ, mobile phase: A: water containing 0.1% formic acid, B: acetonitrile containing 0.1% formic acid, column: poroshell EC-C18, 4.6X105 mm,2.7pm, gradient method: flow: 1.2mL/min, time (min) A (%) B (%)

Time (min)	A(%)	B(%)
			0.00	90	10
1.5	5	95
			2.0	5	95
2.1	90	10
			3.0	90	10

Preparative HPLC was performed on a column (150X 21.2mm ID,5pm,Gemini NXC 18) at a flow rate of 20ml/min, an injection volume of 2ml, at room temperature and UV detection at 214nm and 254 nm.

In the examples below, the following abbreviations are used:

EXAMPLE 1((1S, 3R) -3- (((5-bromo-2-nitrophenyl) amino) methyl) cyclopentyl) methanol

Step1 3-oxabicyclo [3.2.1] octane-2, 4-dione

A2L round bottom flask equipped with a magnetic stirrer was charged with cis-cyclopentane-1, 3-dicarboxylic acid (100 g,633 mmol) and Ac ₂ O (600 ml). The mixture was stirred at 140 ℃ for 12 hours. The mixture was concentrated under reduced pressure. Wet milling the residue with PE to give a crude product (70g,496.4mmol,78.5％).¹H NMR(400MHz,DMSO)δ3.18(q,J＝3.9Hz,2H),2.35(d,J＝12.4Hz,1H),2.17–2.01(m,2H),1.90(t,J＝7.5Hz,2H),1.69(dt,J＝12.4,4.2Hz,1H).

Step 2 cis-3-carbamoyl cyclopentane-1-carboxylic acid

A2L round bottom flask equipped with a magnetic stirrer was charged with (1R, 5S) -3-oxabicyclo [3.2.1] octane-2, 4-dione (70 g,496.4 mmol), THF (700 ml) and NH ₃ -MeOH 7M (4 eq.) at 0deg.C. The mixture was stirred at room temperature for 12 hours. The residue was quenched with MeOH (500 ml) at 0 ℃. The mixture was concentrated under reduced pressure to give a crude product (56 g,354.4mmol, 74.4%). [ M+H ] ⁺ = 158.

Step 3 cis-3-carbamoyl cyclopentane-1-carboxylic acid methyl ester

A1L round bottom flask equipped with a magnetic stirrer was charged with cis-3-carbamoylcyclopentane-1-carboxylic acid (56 g,354.4 mmol) and MeOH (600 mL). The temperature was reduced to 0 ℃ and H ₂SO₄ (34.7 g,354 mmol) was added dropwise to the mixture while maintaining the temperature at 0 ℃. The resulting mixture was stirred at room temperature for 12 hours. The mixture was concentrated under reduced pressure to give the crude product (49 g,284.8mmol, 80.3%). [ M+H ] ⁺ = 172.

Step 4((cis) -3- (aminomethyl) cyclopentyl) methanol

A1L round bottom flask equipped with a magnetic stirrer was charged with cis-3-carbamoylcyclopentane-1-carboxylate (49 g,284.8 mmol) and THF (500 mL). The temperature was reduced to 0 ℃ and BH ₃ -THF (1140 ml,1140 mmol) was added dropwise to the mixture while maintaining the temperature at 0 ℃. The resulting mixture was stirred at room temperature for 24 hours. The residue was quenched with MeOH (500 ml) at 0 ℃. The mixture was concentrated under reduced pressure to give the crude product (35 g,269.2mmol, 79.5%). [ M+H ] ⁺ = 130.

Step 5((cis) -3- (((5-bromo-2-nitrophenyl) amino) methyl) cyclopentyl) methanol

A1L round bottom flask equipped with a magnetic stirrer was charged with ((cis) -3- (aminomethyl) cyclopentyl) methanol (35 g,269.2 mmol), 4-bromo-2-fluoro-1-nitrobenzene (64.8 g,295.9 mmol), DIEA (104.2 g,807.8 mmol) and DMSO (500 mL). The mixture was stirred at 80 ℃ under N ₂ for 4 hours. After cooling to room temperature, the mixture was poured into EA (500 mL), washed successively with brine (200 mL), water (3×200 mL), brine (200 mL), then dried over anhydrous Na ₂SO₄, and concentrated in vacuo. The residue was purified by column chromatography (EA/PE, 24%) to give the title product (20 g,60.8mmol, 22.6%). [ M+H ] ⁺ =329.

Step 6((1S, 3R) -3- (((5-bromo-2-nitrophenyl) amino) methyl) cyclopentyl) methanol

The crude product (20 g) was purified by preparative SFC under the following conditions (((Lux 3u Cellulose-34.6. Mu.m, 50mm,3 μm), temperature: 35 ℃, flow (mL/min): 4, solvent A: CO ₂, solvent B: IPA (0.5% 2mM NH ₃ -MeOH), gradient (B%): 10% to 50% over 2.0min, hold at 50% for 1.0min, hold time 0.971 min) to give ((1S, 3R) -3- (((5-bromo-2-nitrophenyl) amino) methyl) cyclopentyl) methanol (5.6 g,28.1%, ee=96.08%). [ M+H ] ⁺ =329. The crude product can also be purified by chiral HPLC on a column (CHIRALPAK IC-3.6X10 mm,3 μm), mobile phase (Hex (0.1% DEA): etOH=80:20), flow 1.0mL/min, temperature 25 ℃, retention time 2.836min.¹H NMR(300MHz,DMSO-d6)δ8.14(t,J＝5.6Hz,1H),7.98(d,J＝9.1Hz,1H),7.27(d,J＝2.1Hz,1H),6.83(dd,J＝9.1,2.0Hz,1H),4.45(t,J＝5.3Hz,1H),4.34(s,1H),3.43(s,1H),2.23(dt,J＝15.3,7.5Hz,1H),2.11–1.83(m,2H),1.81–1.54(m,1H),1.45–1.20(m,2H),1.06(t,J＝7.0Hz,2H),1.00–0.71(m,1H).

Step 7 methyl 2- (5- (((1S, 3R) -3- (((5-bromo-2-nitrophenyl) amino) methyl) cyclopentyl) methoxy) -1-methyl-1H-pyrazol-4-yl) -6-methyliisonicotinic acid

To a stirred solution of ((1S, 3R) -3- (((5-bromo-2-nitrophenyl) amino) methyl) cyclopentyl) methanol (5.1 g,15.5 mmol), methyl 2- (5-hydroxy-1-methyl-1H-pyrazol-4-yl) -6-methylisonicotinate (4.2 g,17.0 mmol) and PPh ₃ (4.9 g,18.6 mmol) in THF (100 mL) was added DIAD (3.8 g,18.6 mmol). The mixture was then stirred at room temperature for 2 hours. The mixture was then concentrated and purified by column chromatography on silica gel eluting with EtOAc/PE (0-80%) to give the product (9.3 g) mixed with PPh ₃ O, [ m+h ] ⁺ = 558.3.

Step 8 methyl 2- (5- (((1S, 3R) -3- (((2-amino-5-bromophenyl) amino) methyl) cyclopentyl) methoxy) -1-methyl-1H-pyrazol-4-yl) -6-methyliisonicotinic acid

To a stirred solution of methyl 2- (5- (((1 s,3 r) -3- (((5-bromo-2-nitrophenyl) amino) methyl) cyclopentyl) methoxy) -1-methyl-1H-pyrazol-4-yl) -6-methyliisonicotinic acid (9.3 g,16.7 mmol) in THF (100 mL) was added raney nickel (4.5 g). The resulting mixture was stirred at room temperature under a hydrogen atmosphere (1 atm) for 2 hours. The mixture was then filtered and the filtrate was concentrated. The resulting mixture (8.4 g) was used in the next step without purification. [ M+H ] ⁺ = 528.3.

Step 9 methyl 2- (5- (((1S, 3R) -3- ((6-bromo-2-imino-2, 3-dihydro-1H-benzo [ d ] imidazol-1-yl) methyl) cyclopentyl) methoxy) -1-methyl-1H-pyrazol-4-yl) -6-methyliisonicotinic acid methyl ester

A solution of 2- (5- (((1S, 3R) -3- (((2-amino-5-bromophenyl) amino) methyl) cyclopentyl) methoxy) -1-methyl-1H-pyrazol-4-yl) -6-methyliisonicotinic acid methyl ester (8.4 g,15.9 mmol) and BrCN (2.5 g,23.8 mmol) in MeOH (50 mL) was stirred at room temperature for 4 hours. The mixture was then concentrated and diluted with DCM (200 mL), and then washed with saturated aqueous NaHCO ₃ (3×100 mL) and brine (100 mL). The organic layer was dried over anhydrous Na ₂SO₄, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel eluting with MeOH/DCM (0-15%) to give the product (7.1 g, 80.7%); m+h ⁺ =553.4.

Step 10 (7 ¹R,7³S,E)-5⁶ -bromo-1 ¹,2⁶ -dimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazol-2 (2, 4) -pyridin-1 (4, 5) -pyrazole-7 (1, 3) -cyclopentacyclonon-tomato-3-one

To a stirred solution of methyl 2- (5- (((1 s,3 r) -3- ((6-bromo-2-imino-2, 3-dihydro-1H-benzo [ d ] imidazol-1-yl) methyl) cyclopentyl) methoxy) -1-methyl-1H-pyrazol-4-yl) -6-methyliisonicotinic acid (3.2 g,5.8 mmol) in 80mL THF was added LiHMDS (1N in THF, 14.5 mL) at room temperature for 15 min. The resulting mixture was stirred at room temperature for 1 hour. The mixture was then diluted with EA (100 mL), washed with saturated aqueous NH ₄ Cl (2 x 100 mL) and brine (100 mL). The organic layer was dried over anhydrous Na ₂SO₄, filtered and concentrated under reduced pressure. The residue was suspended in 100mL EA and stirred at room temperature for 1 hour. The mixture was then filtered to give the product (2.6 g, 86.2%); m+h ⁺ =521.2.

EXAMPLE 2 (7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-5 ⁶ - (piperazin-1-yl) -5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazole-7 (1, 3) -cyclopentadecalin-3-one

Step 1:4- ((7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridine-1 (4, 5) -pyrazole-7 (1, 3) -cyclopentylnonen-5 ⁶ -yl) piperazine-1-carboxylic acid tert-butyl ester

A solution of example 1 (1.0 g,1.92 mmol), tert-butyl piperazine-1-carboxylate (535 mg,2.88 mmol), pd ₂(dba)₃ (360 mg,0.4 mmol), ruphos (360 mg,0.8 mmol) and NaO ^t Bu (550 mg,5.75 mmol) in DMA (30 mL) was stirred at 100℃for 2 hours. The reaction was concentrated in vacuo and the residue was purified by silica gel column (DCM: CH ₃ oh=15:1) to give tert-butyl 4- ((7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridine-1 (4, 5) -pyrazole-7 (1, 3) -cyclopentylnon-5 ⁶ -yl) piperazine-1-carboxylate (1.0 g, 83.2%) [ m+h ] ⁺ =627.

Step 2 (7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-5 ⁶ - (piperazin-1-yl) -5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazol-2 (2, 4) -pyridin-1 (4, 5) -pyrazole-7 (1, 3) -cyclopentacyclonon-tomato-3-one

To a mixture of tert-butyl 4- ((7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridine-1 (4, 5) -pyrazole-7 (1, 3) -cyclonon-5 ⁶ -yl) piperazine-1-carboxylate (1.0 g,1.6 mmol) in DCM (20 mL) was added TFA (7 mL). The reaction mixture was stirred at room temperature for 2 hours and the resulting mixture was concentrated in vacuo: the combined organic phases were washed with brine (80 mL), dried over Na ₂SO₄, filtered and concentrated in vacuo to give (7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-5 ⁶ - (piperazin-1-yl) -5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridine-1 (4, 5) -pyrazole-7 (1, 3) -cyclopentacyclonon-3-one (700 mg, 83.3%) [ m+h ] ⁺ =527.

Example 3 (7 ¹R,7³S,E)-5⁶ - ((S) -3- (methoxymethyl) piperazin-1-yl) -1 ¹,2⁶ -dimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazol-2 (2, 4) -pyridin-1 (4, 5) -pyrazol-7 (1, 3) -cyclopentylnonen-3-one

The title compound was prepared in a similar manner to example 2. [ m+h ] ⁺ = 571.5

EXAMPLE 4 (7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-5 ⁶ - (2-oxa-5, 8-diazaspiro [3.5] nonan-5-yl) -5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridine-1 (4, 5) -pyrazole-7 (1, 3) -cyclopentadecalin-3-one

The title compound was prepared in a similar manner to example 2. [ m+h ] ⁺ =569.5

Example 5 (7 ¹S,7³R,E)-1¹,2⁶ -dimethyl-5 ⁶ - (piperazin-1-yl) -5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazole-7 (1, 3) -cyclopentadecalin-3-one

The title compound was prepared in a similar manner to examples 1 and 2. [ m+h ] ⁺ =527.5

Example A3- (4- ((3S, 4R) -4- (4- ((7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazole-7 (1, 3) -cyclonon-tomato-5- ⁶ -yl) piperazin-1-yl) -3-fluoropiperidin-1-yl) -3, 3-dimethyl-2-oxoindol-1-yl) piperidine-2, 6-dione

Step 1N- (2, 6-bis (benzyloxy) pyridin-3-yl) -2- (2, 6-dibromophenyl) acetamide

To a solution of 2- (2, 6-dibromophenyl) acetic acid (10 g,34.2 mmol), 2, 6-bis (benzyloxy) pyridin-3-amine (11.5 g,37.6 mml), DIEA (13.3 g,102.6 mmol) in DMF (200 ml) was added HATU (19.5 g,51.3 mmol) at 0 ℃. The mixture was stirred at room temperature overnight. The mixture was diluted with EA (500 mL) and then a solid filtrate was obtained. The mixture was concentrated under reduced pressure to give the crude product (15 g, 75.7%). [ M+H ] ⁺ =581.

Step 2 1- (2, 6-bis (benzyloxy) pyridin-3-yl) -4-bromoindol-2-one

A500 mL round bottom flask equipped with a magnetic stirrer was charged with N- (2, 6-bis (benzyloxy) pyridin-3-yl) -2- (2, 6-dibromophenyl) acetamide (15 g,25.9 mmol), ((2-bromoethoxy) methyl) benzene (15.4 g,71.5 mmol), K ₂CO₃ (17.8 g,129.5 mmol), cuCl (2.56 g,25.9 mmol), pentane-2, 4-dione (5.17 g,51.8 mmol) and NMP (200 mL). The mixture was degassed under vacuum and purged three times with N ₂. The resulting mixture was stirred at 85 ℃ under N ₂ for 3 hours. After cooling to room temperature, the mixture was diluted with ethyl acetate (300 mL) and then filtered through a pad of celite. The filtrate was washed with brine (300 mL), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA/PE, 15%) to give the product (8.00 g, 61.8%). [ M+H ] ⁺ =501.

Step 3 4- ((3S, 4R) -4- (4-Benzylpiperazin-1-yl) -3-fluoropiperidin-1-yl) -1- (2, 6-bis (benzyloxy) pyridin-3-yl) indolin-2-one

A solution of 1- (2, 6-bis (benzyloxy) pyridin-3-yl) -4-bromoindolin-2-one (1 g,2 mmol), 1-benzyl-4- ((3S, 4R) -3-fluoropiperidin-4-yl) piperazine (381 mg,3mmol, prepared in a similar manner to that in WO2023098656A 1), ^t- Buona (576 mg,6 mmol), pd-PEPSI-IPentCl (195 mg,0.2 mmol) in DMA (15 mL) was degassed under reduced pressure and purged five times with N ₂ and stirred at 90℃for 1 hour under N ₂. After cooling to room temperature, the mixture was diluted with DCM (100 mL) and filtered through a pad of celite. The filtrate was washed with brine (150 mL), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA/PE, 40%) to give the product (810 mg, 60%). [ M+H ] ⁺ = 698.3.

Step 4:4- ((3S, 4R) -4- (4-Benzylpiperazin-1-yl) -3-fluoropiperidin-1-yl) -1- (2, 6-bis (benzyloxy) pyridin-3-yl) -3, 3-dimethylindolin-2-one

DMF (10 mL) containing 4- ((3 s,4 r) -4- (4-benzylpiperazin-1-yl) -3-fluoropiperidin-1-yl) -1- (2, 6-bis (benzyloxy) pyridin-3-yl) indolin-2-one (697 mg,1 mmol) and stirring at 0 ℃ under N ₂. NaH (100 mg,2.5mmol,60% in oil) was added at this temperature. After 30 minutes, CH ₃ I (355 mg,2.5 mmol) was added and stirred at room temperature under N ₂ for 1 hour. The reaction solution was quenched with 20mL of water and extracted with DCM. The organic layer was washed with brine, saturated aqueous sodium thiosulfate solution and water, and dried over anhydrous sodium sulfate. It was filtered and the resulting residue was concentrated, which was purified by using column chromatography (EA/PE, 40%) to give the product (500 mg, 69%). [ m+h ] += 726.5.

Step 5:4- ((3S, 4R) -1- (1- (2, 6-dioxopiperidin-3-yl) -3, 3-dimethyl-2-oxoindol-4-yl) -3-fluoropiperidin-4-yl) piperazine-1-carboxylic acid tert-butyl ester

To a solution of 4- ((3 s,4 r) -4- (4-benzylpiperazin-1-yl) -3-fluoropiperidin-1-yl) -1- (2, 6-bis (benzyloxy) pyridin-3-yl) -3, 3-dimethylindol-2-one (726 mg,1 mmol) in DMF/^i- pro h (5 mL/5 mL) was added 10% Pd/C (100 mg) and (Boc) ₂ O at room temperature under N ₂. And then the mixture was exchanged with H ₂ twice and stirred under an atmosphere of H ₂ at 50 ℃ for 15 hours. The mixture was filtered through a pad of celite and washed with MeOH (50 mL). The filtrate was concentrated in vacuo to give the product (500 mg, 90%) [ m+h ] ⁺ = 558.5.

Step 63- (4- ((3S, 4R) -3-fluoro-4- (piperazin-1-yl) piperidin-1-yl) -3, 3-dimethyl-2-oxoindolin-1-yl) piperidine-2, 6-dione

To a solution of 4- ((3 s,4 r) -1- (1- (2, 6-dioxopiperidin-3-yl) -3, 3-dimethyl-2-oxoindol-4-yl) -3-fluoropiperidin-4-yl) piperazine-1-carboxylic acid tert-butyl ester (279 mg,0.5 mmol) in DCM (3 mL) was added TFA (1 mL). The reaction was stirred at room temperature for 2 hours, and then concentrated in vacuo. The residue was dissolved in DCM (20 mL), washed with saturated NaHCO ₃ solution (3×20 mL) and brine (50 mL), dried over Na ₂SO₄, filtered and concentrated under reduced pressure to give the product (180 mg, 78.6%); m+h ] += 458.5.

Step 7 3- (4- ((3S, 4R) -4- (4- ((7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazol-7 (1, 3) -cyclopentylnon-tomato-5- ⁶ -yl) piperazin-1-yl) -3-fluoropiperidin-1-yl) -3, 3-dimethyl-2-oxoindol-1-yl) piperidine-2, 6-dione

A mixture of example 1 (104 mg,0.2 mmol), 3- (4- ((3S, 4R) -3-fluoro-4- (piperazin-1-yl) piperidin-1-yl) -3, 3-dimethyl-2-oxoindolin-1-yl) piperidine-2, 6-dione (137 mg,0.3 mmol), pd ₂dba₃ (36.6 mg,0.04 mmol), ruphos (36.8 mg,0.08 mmol) and ^t- Buona (76.8 mg,0.8 mmol) in DMA (5 mL) was stirred in a round bottom flask at 90℃under N ₂ for 1 hour. Water (10 mL) was added and the mixture extracted with DCM (20 mL. Times.3). The combined organic layers were dried over Na ₂SO₄. The solvent was removed by evaporation and the residue was purified by column chromatography on silica gel to give the product (40mg,22％).¹H NMR(500MHz,DMSO)δ12.43(s,1H),11.07(s,1H),8.53(s,1H),7.89(s,1H),7.45(s,1H),7.38(d,J＝8.7Hz,1H),7.25(t,J＝8.0Hz,1H),7.10–7.00(m,2H),6.95–6.90(m,1H),6.83(s,1H),5.25–5.21(m,1H),5.15–5.06(m,1H),4.38–4.25(m,1H),4.22–4.16(m,1H),4.14–4.02(m,2H),3.74(s,3H),3.24–3.17(m,4H),3.15–2.97(m,2H),2.96–2.80(m,7H),2.64–2.54(m,9H),2.09–1.99(m,1H),1.98–1.76(m,6H),1.69–1.65(m,1H),1.43(dd,J＝8.3,3.8Hz,6H);[M+H]⁺＝898.6.

EXAMPLE B (R) -3- (4- (4- ((7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazole-7 (1, 3) -cyclopentanecyclononan-5 ⁶ -yl) piperazin-1-yl) -piperidin-1-yl) -2, 6-difluorophenyl) piperidine-2, 6-dione

Step 12, 6-bis (benzyloxy) -3- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyridine

The 4 reactions were performed in parallel.

To a solution of 2, 6-dibenzyloxy-3-bromo-pyridine (2.00 kg,5.40mol,1.00 eq.) in dioxane (12.0L) was added 4, 5-tetramethyl-1, 3, 2-dioxaborane (760 g,5.94mol,862ml,1.10 eq.) and TEA (1.09 kg,10.8mol,1.50L,2.00 eq.) were degassed and purged with N ₂, then Pd (PPh ₃)₂Cl₂ (189 g,270mmol,0.05 eq.) was added to the mixture, the mixture was stirred at 90 ℃ for 16 hours, the reaction mixture was filtered and concentrated, the 4 reactions were combined, the residue was purified by column chromatography (SiO ₂, petroleum ether/ethyl acetate=1/0 to 0/1) to give 2, 6-bis (benzyloxy) -3- (4, 5-tetramethyl-1, 3, 2-dioxaborane-3.80 kg [ 9.80M ] pyridine (3.37.35% h=3.37.13.80 mol)

Step 2 5-bromo-1, 3-difluoro-2-iodobenzene

The 5 reactions were performed in parallel.

LDA (2M, 2.59L) was added to a solution of 1-bromo-3, 5-difluoro-benzene (1.00 kg,5.18mol,595 mL) in THF (4.00L) at-70 ℃. The mixture was stirred at-70 ℃ for 1 hour. A solution of I ₂ (1.33 kg,5.23mol, 1.05L) in THF (1.00L) was then added to the mixture at-70 ℃. The mixture was stirred at-70 ℃ for 1 hour. The reaction mixture was poured into H ₂ O (5.00L), extracted with EtOAc (3.00 l×2), the 5 reactions combined, the combined organic layers washed with brine (5.00L), dried over Na ₂SO₄, filtered and concentrated in vacuo at 40 ℃ to give a residue. The crude product was wet-milled with petroleum ether (6.00L). The compound 5-bromo-1, 3-difluoro-2-iodobenzene (4.50 kg,14.1mol,54.4% yield) was obtained. [ M+H ] ⁺ = 318.8.

Step 32, 6-bis (benzyloxy) -3- (4-bromo-2, 6-difluorophenyl) pyridine

To a solution of 5-bromo-1, 3-difluoro-2-iodobenzene (4.50 kg,14.1 mol) and 2, 6-bis (benzyloxy) -3- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyridine (5.89 kg,14.1 mol) in dioxane (22.5L) and H ₂ O (4.50L) was added Pd (PPh ₃)₄(1.63kg,1.41mol)、K₃PO₄ (8.99 kg,42.3 mol.) the mixture was stirred at 90 ℃ for 12 hours, the mixture was filtered and the filtrate was extracted with EtOAc (5.00L), the combined organics were washed with brine (5.00L), dried over Na ₂SO₄, and the residue was purified by column chromatography (SiO ₂, petroleum ether: ethyl acetate=1:0 to 1:1:1.) to give the compound 2, 6-bis (benzyloxy) -3- (4-bromo-2, 6-difluorophenyl) pyridine (3.99 mol., 6.00 mol.) in a purity of 96.01.3% yield, 96.00 mol% ).¹H NMR(400MHz,CHLOROFORM-d)7.47-7.40(m,1H),7.39-7.34(m,2H),7.34-7.29(m,2H),7.29-7.25(m,4H),7.25-7.17(m,2H),7.12-7.05(m,2H),6.42(d,J＝8.0Hz,1H),5.32(s,2H),5.28(s,2H);[M+H]⁺＝482.1.

Step 4 8- (4- (2, 6-bis (benzyloxy) pyridin-3-yl) -3, 5-difluorophenyl) -1, 4-dioxa-8-azaspiro [4.5] decane

To a solution of 2, 6-bis (benzyloxy) -3- (4-bromo-2, 6-difluorophenyl) pyridine (30 g,62.24 mmol), 1, 4-dioxa-8-azaspiro [4.5] decane (10.68 g,74.69 mmol) and Cs ₂CO₃ (40.58 g,124.48 mmol) in 500mL dioxane was added Pd ₂(dba)₃ (2.85 g,3.11 mmol) and Xantphos (3.6 g,6.22 mmol) under an N ₂ atmosphere. The mixture was stirred under protection of N ₂ at 80℃for 16 hours. The mixture was diluted with EtOAc and filtered. The filtrate was concentrated in vacuo and purified by silica gel column chromatography (EA: pe=0-80%) to give the crude product. The crude material was recrystallized from MeOH and filtered. The filter cake was dried to give the title compound (26.8 g,79% yield); [ m+h ] ⁺ = 544.9.

Step 5 3- (2, 6-difluoro-4- (1, 4-dioxa-8-azaspiro [4.5] dec-8-yl) phenyl) piperidine-2, 6-dione

To a solution of 8- (4- (2, 6-bis (benzyloxy) pyridin-3-yl) -3, 5-difluorophenyl) -1, 4-dioxa-8-azaspiro [4.5] decane (26.8 g,49.26 mmol) in 400mL DMF and 80mL iPrOH was added Pd/C (27 g,10wt.%, wet). The mixture was stirred at 45 ℃ under an atmosphere of H ₂ (4 bar) for 16 hours. The mixture was filtered and the filter cake was washed with DMF. The combined liquids were concentrated in vacuo to give the title compound (15 g,83.2% yield); [ m+h ] ⁺ = 367.1.

Step 6 (R) -3- (2, 6-difluoro-4- (1, 4-dioxa-8-azaspiro [4.5] dec-8-yl) phenyl) piperidine-2, 6-dione

The title compound was purified by chiral HPLC (CHIRALPAK IF (2 x 25cm,5 um), mtBE (0.1% dea): (MeOH: dcm=1:1) =50:50, 100 bar, 20 ml/min) and corresponds to peak a at 0.990min/254nm (4.47 g,12g racemate yield 37%); m+h ] ⁺ = 367.1.

Step 7 (R) -3- (2, 6-difluoro-4- (4-oxopiperidin-1-yl) phenyl) piperidine-2, 6-dione

(R) -3- (2, 6-difluoro-4- (1, 4-dioxa-8-azaspiro [4.5] dec-8-yl) phenyl) piperidine-2, 6-dione (1 g,2.73 mmol) was placed in a 100mL round bottom flask with a magnetic stirring rod. Then, 10mL of 8N aqueous HCl was added. The mixture was stirred at room temperature for 30 minutes. The mixture was added dropwise to a saturated aqueous solution of NaHCO ₃, and finally, ph=6-7. The liquid was extracted with DCM and separated. The organic phase was concentrated in vacuo and purified by silica gel column chromatography (MeOH: dcm=0-5%) to give the title compound (850 mg,96.7% yield); [ m+h ] ⁺ =323.1.

Step 8 (R) -3- (4- (4- (4- ((7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazol-7 (1, 3) -cyclopentanecyclononan-5 ⁶ -yl) piperazin-1-yl) -2, 6-difluorophenyl) piperidin-2, 6-dione

To a solution of example 2 (80 mg,0.15 mmol) and (R) -3- (2, 6-difluoro-4- (4-oxopiperidin-1-yl) phenyl) piperidine-2, 6-dione (68 mg,0.21 mmol) in DCE (8 mL) was added STAB (95 mg,0.45 mmol). The mixture was then stirred at 50 ℃ overnight. The reaction was quenched with saturated aqueous NaHCO ₃ and extracted with DCM (3×15 m). The combined organic phases were washed with brine (1×10 mL), dried over Na ₂SO₄, filtered and concentrated in vacuo. The residue was purified by a silica gel column (DCM: CH ₃ oh=10:1) followed by purification by preparative HPLC chromatography to give the title product (33mg,26.1％).¹H NMR(500MHz,DMSO)δ12.42(s,1H),10.87(s,1H),8.52(s,1H),7.89(s,1H),7.45(s,1H),7.36(d,J＝8.7Hz,1H),7.01(s,1H),6.90(d,J＝8.7Hz,1H),6.66(s,1H),6.63(s,1H),4.30(s,1H),4.19(s,1H),4.05(dd,J＝12.5,5.0Hz,3H),3.81(d,J＝12.1Hz,2H),3.74(s,3H),3.16(s,4H),2.77(dd,J＝15.1,7.7Hz,3H),2.68(s,4H),2.65–2.57(m,2H),2.55(s,3H),2.53(d,J＝4.0Hz,2H),2.49–2.40(m,2H),2.09(qd,J＝13.4,4.9Hz,1H),1.99–1.79(m,6H),1.67(s,1H),1.54–1.43(m,2H).[M+H]⁺＝833.7

EXAMPLE C5- (3- (((S) -4- ((7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazole-7 (1, 3) -cyclonon-5- ⁶ -yl) -2- (methoxymethyl) piperazin-1-yl) methyl) azetidin-1-yl) -2- (2, 6-dioxopiperidin-3-yl) isoindoline-1, 3-dione

Step 12- (2, 6-Dioxopiperidin-3-yl) -5- (3- (hydroxymethyl) azetidin-1-yl) isoindoline-1, 3-dione

A mixture of 2- (2, 6-dioxopiperidin-3-yl) -5-fluoroisoindoline-1, 3-dione (2.76 g,1 mmol), azetidin-3-ylmethanol (870 mg,1 mmol) and DIEA (2.85 g,2 mmol) in DMSO (30 mL) was stirred in a round bottom flask at 100℃under N ₂ for 12 hours. The mixture was diluted with H ₂ O and extracted with EtOAc (100 ml×3). The combined organic layers were washed with brine (100 mL) and dried over anhydrous Na ₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel eluting with CH ₂Cl₂/MeOH (10:1). 2- (2, 6-Dioxopiperidin-3-yl) -5- (3- (hydroxymethyl) azetidin-1-yl) isoindoline-1, 3-dione (3 g, 87%) was obtained. [ m+h ] ⁺ = 344.3

Step 2 1- (2, 6-Dioxopiperidin-3-yl) -1, 3-dioxoisoindolin-5-yl) azetidine-3-carbaldehyde

A mixture of 2- (2, 6-dioxopiperidin-3-yl) -5- (3- (hydroxymethyl) azetidin-1-yl) isoindoline-1, 3-dione (1.71 g,0.5 mmol), 2-iodooxybenzoic acid (2.8 g,1 mmol) in DMSO (15 mL) was stirred in a round bottom flask at 25℃under an atmosphere of N ₂ for 12 hours. The mixture was diluted with H ₂ O and extracted with DCM (100 ml×3). The combined organic layers were washed with brine (100 mL) and dried over anhydrous Na ₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel eluting with CH ₂Cl₂/MeOH (10:1) to give the title compound (1.5 g, 88%). [ m+h ] ⁺ = 342.3 step 4:5- (3- (((S) -4- ((7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazole-7 (1, 3) -cyclopentylnon-tomato-5 ⁶ -yl) -2- (methoxymethyl) piperazin-1-yl) methyl) azetidin-1-yl) -2- (2, 6-dioxopiperidin-3-yl) isoindoline-1, 3-dione

To a solution of example 3 (85 mg,0.15 mmol) and 1- (2, 6-dioxopiperidin-3-yl) -1, 3-dioxoisoindolin-5-yl) azetidine-3-carbaldehyde (72 mg,0.21 mmol) in DCE (6 mL) was added STAB (96 mg,0.45 mmol). The mixture was then stirred at room temperature overnight. The reaction was quenched with saturated aqueous NaHCO ₃ (15 mL) and extracted with DCM (3×15 mL). The combined organic phases were washed with brine (1×10 mL), dried over Na ₂SO₄, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography (DCM: CH ₃ oh=10:1) followed by preparative HPLC chromatography to give the title product (60 mg, 45%).

¹H NMR(500MHz,DMSO)δ12.55(s,1H),11.08(s,1H),8.61(s,1H),7.99(s,1H),7.70(d,J＝8.3Hz,1H),7.57(s,1H),7.43(d,J＝8.7Hz,1H),7.13(d,J＝17.3Hz,1H),7.00(t,J＝7.5Hz,1H),6.84(d,J＝1.6Hz,1H),6.71(dd,J＝8.4,1.9Hz,1H),5.07(dd,J＝12.7,5.4Hz,1H),4.37–4.18(m,5H),4.15–4.04(m,3H),3.88–3.81(m,10H),3.76–3.72(m,3H),3.69–3.62(m,2H),3.58–3.51(m,1H),3.48–3.42(m,1H),3.32–3.21(m,2H),3.09–2.99(m,2H),2.94–2.84(m,1H),2.64–2.55(m,6H),2.06–1.99(m,1H),1.93–1.88(m,1H),1.84–1.79(m,2H),1.68–1.59(m,1H);[M+H]⁺＝896.7.

EXAMPLE D (R) -3- (4- (4- (5- ((7 ¹R,7³S,E)-1¹,2⁶ -dimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-9-oxa-4-aza-5 (2, 1) -benzo [ D ] imidazole-2 (2, 4) -pyridine-1 (4, 5) -pyrazole-7 (1, 3) -cyclopentanecyclononan-5 ⁶ -yl) -2-oxa-5, 8-diazaspiro [3.5] nonan-8-yl) piperidin-1-yl) -2, 6-difluorophenyl) piperidine-2, 6-dione

The title compound was prepared in analogy to example B using example 4 and (R) -3- (2, 6-difluoro-4- (4-oxopiperidin-1-yl) phenyl) piperidine-2, 6-dione .¹H NMR(500MHz,DMSO)δ12.50(s,1H),10.87(s,1H),8.52(s,1H),7.89(s,1H),7.45(s,1H),7.40(d,J＝8.5Hz,1H),7.02(s,1H),6.75(dd,J＝8.5,1.6Hz,1H),6.66(s,1H),6.63(s,1H),4.53(d,J＝6.1Hz,2H),4.38(d,J＝6.2Hz,2H),4.35–4.27(m,1H),4.19(t,J＝8.2Hz,1H),4.10(t,J＝8.4Hz,1H),4.05(dd,J＝12.6,5.1Hz,2H),3.80(d,J＝12.5Hz,2H),3.74(s,3H),3.26–3.18(m,2H),2.99(s,2H),2.84–2.74(m,3H),2.65–2.51(m,7H),2.49–2.46(m,4H),2.09(qd,J＝13.0,3.7Hz,1H),2.00–1.89(m,2H),1.83(d,J＝10.1Hz,4H),1.75–1.66(m,1H),1.56–1.46(m,2H).[M+H]⁺＝875.7.

Cellular p-EGFR inhibition assay

Cell treatment

On day 1, H1975-clone #23 (DEL 19/C797S) cells were seeded at 3X 10 ⁴ cells/well in cell culture medium [ RPMI1640 (Gibco, catalog No. 72400-047), 10% heat-inactivated FBS,1% PS (Gibco, catalog No. 10378) ] in Corning 96 well plates (catalog No. 3599).

On day 2, baF3-L858R cells were seeded at 2×10 ⁵ cells/well in cell culture medium [ RPMI1640 (Gibco, without phenol red, catalog No. 11835-030), 10% heat-inactivated FBS,1% PS (Gibco, catalog No. 10378) ] in Corning 96-well plates (catalog No. 3799) at a volume of 54 μl/well.

H1975- #23 and BaF3-L858R cells were treated with compounds diluted in 0.1% DMSO cell culture on day 2, incubated for 16H,37℃at 5% CO ₂. The final concentration of compound in all assays was from 10uM, 5-fold dilution, total 8 doses.

HTRF assay

After 16H treatment, 50 μl HTRF 1 Xlysis buffer was added to each well for H1975- #23 cells, 20 μl 4 Xlysis buffer was added to each well for BaF3-L858R cells, the plates were sealed and incubated for 1 hour at room temperature on a plate shaker, 16 μl of cell lysate was transferred to PE 384 well HTRF assay plates once the cells were lysed, 4 μl of premixed HTRF antibody was added to each well, the plates were covered with plate sealer, spun at 1000rpm for 1min, incubated overnight at room temperature, and read on BMG PheraStar using HTRF protocol (337 nm-665nm-620 nm).

The percent inhibition of the compounds was calculated by the following equation, percent inhibition of the compounds = 100-100× (signal-low control)/(high control-low control), where signal = low control per test compound group = lysis buffer alone, cell free, indicating complete inhibition of p-EGFR.

High control = DMSO-added and compound-free cell group, indicating no p-EGFR inhibited microplate reading.

Imax is the percent maximum inhibition.

The IC ₅₀ value for a compound can be obtained by fitting the following equation:

Y=floor+ (top-bottom)/(1+ ((IC ₅₀/X)/(schl slope))

Where X and Y are known values, and IC ₅₀, hill slope (Hillslope), top and bottom values are parameters obtained by fitting with software. Y is the percent inhibition (calculated from the equation), X is the concentration of the compound, and IC ₅₀ is the concentration of the compound at which 50% inhibition is achieved. The smaller the IC ₅₀ value, the more inhibitory the compound is. Conversely, the higher the IC ₅₀ value, the weaker the inhibition of the compound, the slope of the hill represents the slope of the fitted curve, typically about 1 x, the bottom represents the minimum of the curve obtained by data fitting, typically 0% ± 20%, and the top represents the maximum of the curve obtained by data fitting, typically 100% ± 20%. Experimental data were fitted by calculation and analysis with Dotmatics data analysis software.

TABLE 1 degradation (BaF 3-L858R and H1975-clone #8 (L858R/C797S)) results of examples

The foregoing examples and description of certain embodiments should be regarded as illustrative rather than limiting the invention as defined by the claims. It will be readily appreciated that numerous variations and combinations of the features described above may be utilized without departing from the present invention as set forth in the claims. All such variations are intended to be included within the scope of the present invention. All cited references are incorporated herein by reference in their entirety.

It will be understood that, if any prior art publication is referred to herein, such reference does not constitute an admission that the publication forms a part of the common general knowledge in the art in any country.

Claims

1. A compound of formula (I):

in:

R ^1a , R ^1b , R ^2a and R ^2b are each independently absent, hydrogen, halogen, -C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, -C _1-8 alkoxy, -C _3-8 cycloalkyl or -CN; each of the -C _1-8 alkyl, -C _2-8 alkenyl, -C _2-8 alkynyl, -C _1-8 alkoxy or -C _3-8 cycloalkyl is optionally substituted with at least one substituent selected from hydrogen, halogen, -C _1-8 alkoxy, -C _3-8 cycloalkyl or -CN;

R ³ and R ⁴ are each independently hydrogen, -C _1-6 alkyl or -C _3-8 cycloalkyl; each of the -C _1-6 alkyl or -C _3-8 cycloalkyl is optionally substituted by at least one substituent selected from hydrogen, halogen, -C _1-6 alkoxy;

R ⁷ is each independently absent, hydrogen, halogen, -C _{1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, -C 1-8 alkoxy, -C 3-8 cycloalkyl or -CN; each of said -C 1-8} _{alkyl, -C 2-8} _alkenyl _, _-C _2-8 _alkynyl _, -C _1-8 alkoxy or -C _3-8 cycloalkyl is optionally substituted with at least one substituent selected from hydrogen, halogen, -C _1-8 alkoxy, -C _3-8 cycloalkyl or -CN; or

Two R ⁷ together with the carbon atom to which they are attached form a 3- to 12-membered ring, the ring comprising 0-3 heteroatoms independently selected from nitrogen, oxygen or sulfur; the ring is optionally substituted with at least one substituent halogen, hydroxyl or -C ₁ -C ₈ alkyl;

R ⁸ and R ⁹ are each independently selected from hydrogen, halogen, -C ₁ -C ₆ alkyl or C ₃ -C ₈ cycloalkyl; -C ₁ -C ₆ alkyl or C ₃ -C ₈ cycloalkyl are each optionally substituted with at least one substituent selected from hydrogen, halogen, -C _1-6 alkoxy;

^R10 is independently selected from halogen, It is stated

each is optionally substituted with at least one R ^10a ;

The R ^10a are each independently absent, oxo (=O), halogen, hydroxyl, -CN, -C ₁ -C ₈ alkyl, -C ₁ -C ₈ alkoxy, -C ₂ -C _{8 alkenyl, -C 2 -C 8 alkynyl, C 3 -C 8} _cycloalkyl _, ₃ _to 8 membered heterocyclyl, C ₆ -C ₁₂ aryl, or 5 to 12 membered heteroaryl; the -C ₁ -C ₈ alkyl, -C ₁ -C ₈ alkoxy, -C ₂ -C ₈ alkenyl, -C ₂ -C ₈ alkynyl, C ₃ -C ₈ cycloalkyl, 3 to 8 membered heterocyclyl, C ₆ -C ₁₂ aryl, and 5 to 12 membered heteroaryl are each optionally substituted by at least one R ^10b ,

R ^10b is independently absent, oxo (═O), halogen, hydroxy, —CN, —C ₁ -C ₈ alkyl, —C ₁ -C ₈ alkoxy, —C ₂ -C ₈ alkenyl, —C ₂ -C ₈ alkynyl, C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl, C ₆ -C ₁₂ aryl, or 5- to 12-membered heteroaryl; or

Two R ^10a together with the atoms to which they are attached form a 3- to 12-membered ring, the ring comprising 0-3 heteroatoms independently selected from nitrogen, oxygen or sulfur; the ring is optionally substituted with at least one substituent halogen, hydroxyl, -C ₁ -C ₈ alkyl;

At each occurrence, ^X1 is independently selected from ^-CRa or N;

At each occurrence, ^X2 is independently selected from -NH- or -C(O)-;

At each occurrence, ^X3 is independently selected from ^-NRa- , -O-, -S- and ^-CRaRb- ^;

At each occurrence, ^Ra and ^Rb are each independently selected from hydrogen, hydroxy, halogen, CN, -Ci _- _C8 alkyl, -Ci _- _C8 alkoxy, _-C2 - _C8 alkenyl, -C2 _- C8 alkynyl, _{-C3-C8 cycloalkyl, 3 to 8 membered heterocyclyl, -C6-C12 aryl or 5 to 12 membered heteroaryl, wherein the -Ci-C8 alkyl, -Ci-C8 alkoxy, -C2-C8 alkenyl, -C2-C8 alkynyl, -C3-C8 cycloalkyl, 3} _to ₈ _membered _heterocyclyl _, _-C6 _- _C12 _aryl _or ₅ _to ₁₂ _membered _heteroaryl are each optionally substituted by at least one halogen, hydroxy, halogen, -Ci- _C8 alkyl, -Ci- _C8 alkoxy, _-C2 -C8 alkenyl, _-C2 - _C8 alkynyl, -C3-C8 cycloalkyl, ₃ to ₈ membered heterocyclyl, -C6-C12 aryl or 5 _to 12 membered heteroaryl. _8- membered alkynyl, -C ₃ -C ₈ cycloalkyl, 3- to 8-membered heterocyclyl, -C ₆ -C ₁₂ aryl, or 5- to 12-membered heteroaryl; or

^Ra and ^Rb together with the carbon atoms to which they are attached form a 3- to 12-membered ring comprising 0-3 heteroatoms independently selected from nitrogen, oxygen or sulfur; the ring is optionally substituted with at least one substituent selected from halogen, hydroxy, _-C1 - _C8 alkyl, _-C2 - _C8 alkenyl, _-C2 - _C8 alkynyl, _-C1 _- _C8 alkoxy, _-C2 - _C8 alkenyl, -C2- _C8 alkynyl, _C3 - _C8 cycloalkyl, 3- to 8-membered heterocyclyl, _C6 - _C12 aryl or 5- to 12-membered heteroaryl;

n1, n2, n3, n4 and n5 are each independently 0, 1, 2 or 3;

s5, s6 and s7 are each independently 0, 1, 2 or 3.

2. The compound according to claim 1, wherein the compound is selected from the group consisting of formula (IIa),

Preferably, the compound is selected from formula (IIb) and (IIc):

Preferably, the compound is selected from formula (IId) and (IIe):

Preferably, the compound is selected from formula (IIf) and (Ig):

wherein R ³ , R ⁴ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , s5 , s6 and s7 are as defined in claim 1 .

3. A compound as claimed in any one of the preceding claims, wherein R ^1a , R ^1b , R ^2a and R ^2b are each independently hydrogen, F, Cl, Br, I, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, -C _2-8 alkenyl, -C _2-8 alkynyl, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, heptyloxy, octyloxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl or -CN; wherein each of the methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, -C _2-8 alkenyl, -C 2-8 alkynyl, _2-8 alkynyl, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, heptyloxy, octyloxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl are optionally substituted with at least one substituent selected from hydrogen, F, Cl, Br, I, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, heptyloxy, octyloxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl or -CN;

Preferably, R ^1a , R ^1b , R ^2a and R ^2b are each independently hydrogen, F, Cl, Br, I, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyl, heptyloxy, octyloxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, -CF ₃ , -CHF ₂ , -CN, -CH ₂ OCH ₃ , -CH ₂ OCH ₂ CH ₃ , -CH ₂ CH ₂ OCH ₃ , -CH ₂ CH ₂ OCH ₂ CH ₃ ;

More preferably, R ^1a , R ^1b , R ^2a and R ^2b are each independently hydrogen, F, Cl, methyl, methoxy, cyclopropyl, -CF ₃ or -CHF ₂ , -CH ₂ OCH ₃ ;

More preferably, R ^1a , R ^1b , R ^2a and R ^2b are each independently hydrogen.

4. A compound as claimed in any one of the preceding claims, wherein R ³ and R ⁴ are each independently hydrogen, methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl; said methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl being optionally substituted with at least one substituent selected from hydrogen, F, Cl, Br, I, methoxy, ethoxy, propoxy, butoxy, pentyloxy or hexyloxy;

Preferably, R ³ and R ⁴ are each independently hydrogen, methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl; preferably, R ³ is independently methyl and R ⁴ is hydrogen.

5. A compound as claimed in any one of the preceding claims, wherein each R ⁷ is independently hydrogen, F, Cl, Br, I, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyl, heptyloxy, octyloxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, -CF ₃ , -CHF ₂ , -CN, -CH ₂ OCH ₃ , -CH ₂ OCH ₂ CH ₃ , -CH ₂ CH ₂ OCH ₃ , -CH ₂ CH ₂ OCH ₂ CH ₃ ;

Preferably, R ⁷ is each independently hydrogen, F, Cl, methyl, methoxy, cyclopropyl, -CF ₃ or -CHF ₂ , -CH ₂ OCH ₃ .

6. A compound as claimed in any one of the preceding claims, wherein two R ⁷ and the carbon atom to which they are attached form a 3, 4, 5, 6, 7 or 8 membered ring comprising 0-3 heteroatoms independently selected from nitrogen, oxygen or sulfur; the ring is optionally substituted with at least one substituent F, Cl, Br, I, hydroxyl, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl or octyl;

Preferably, two R ⁷ and the carbon atom to which they are attached form a 3-, 4-, 5- or 6-membered ring, which is optionally substituted with at least one substituent F, Cl, Br, I, hydroxy, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl or octyl.

7. A compound as claimed in any one of the preceding claims, wherein R ⁸ and R ⁹ are each independently selected from hydrogen, F, Cl, Br, I, methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl; said methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl being optionally substituted with at least one substituent selected from hydrogen, F, Cl, Br, I, methoxy, ethoxy, propoxy, butoxy, pentyloxy or hexyloxy;

Preferably, ^R8 and ^R9 are each independently hydrogen, F, Cl, Br, I, methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl; preferably, ^R8 is independently hydrogen and ^R9 is F or methyl.

8. A compound as claimed in any one of the preceding claims, wherein R ¹⁰ is each independently selected from hydrogen, -F, -Cl, -Br, -I,

It is stated each is optionally substituted with at least one R ^10a ;

The R ^10a are each independently hydrogen, oxo (=O), F, Cl, Br, I, hydroxyl, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyl, heptyloxy, octyloxy, -C ₂ -C ₈ alkenyl, -C ₂ -C ₈ alkynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 3 to 8 membered heterocyclyl, phenyl or 5 to 12 membered heteroaryl; the methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, heptyloxy, octyloxy, -C ₂ -C ₈ alkenyl, -C ₂ -C 8 R 10b is substituted with at _least one R ^10b ,

R ^10b is independently oxo (═O), F, Cl, Br, I, hydroxy, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, heptyloxy, octyloxy, —C ₂ -C ₈ alkenyl, —C ₂ -C ₈ alkynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 3- to 8-membered heterocyclyl, phenyl, or 5- to 12-membered heteroaryl; or

Two R ^10a together with the carbon atom to which they are attached form a 3, 4, 5, 6, 7 or 8 membered ring comprising 0, 1, 2 or 3 heteroatoms independently selected from nitrogen, oxygen or sulfur; the ring is optionally substituted with at least one substituent F, Cl, Br, I, hydroxy, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl or octyl;

Preferably, R ¹⁰ is independently selected from hydrogen, -F, -Cl, -Br, -I,

9. A compound as claimed in any one of the preceding claims, selected from