Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
  • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/50—Pyridazines; Hydrogenated pyridazines
  • A61K31/501—Pyridazines; Hydrogenated pyridazines not condensed and containing further heterocyclic rings
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/50—Pyridazines; Hydrogenated pyridazines
  • A61K31/5025—Pyridazines; Hydrogenated pyridazines ortho- or peri-condensed with heterocyclic ring systems
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
  • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
  • C07D471/04—Ortho-condensed systems

Definitions

• the present disclosure relates to compounds comprising a Bcl-xL inhibiting moiety covalently linked to a degradation signaling moiety (DSM) that binds to a degradation protein or degradation protein complex, e.g., an E3 ubiquitin ligase or an E3 ubiquitin ligase complex.
• DSM degradation signaling moiety
• the disclosure further relates to methods and compositions useful in the treatment of cancers that are responsive to decrease in Bcl-xL expression and/or activity.
• Apoptosis (programmed cell death) is an evolutionarily conserved pathway essential for tissue homeostasis, development and removal of damaged cells.
• Deregulation of apoptosis contributes to human diseases, including malignancies, neurodegenerative disorders, diseases of the immune system and autoimmune diseases (Hanahan and Weinberg, Cell. 2011 Mar 4;144(5):646-74; Marsden and Strasser, Annu Rev Immunol.2003;21:71-105; Vaux and Flavell, Curr Opin Immunol.2000 Dec;12(6):719-24).
• Evasion of apoptosis is recognized as a hallmark of cancer, participating in the development as well as the sustained expansion of tumors and the resistance to anti-cancer treatments (Hanahan and Weinberg, Cell.2000 Jan 7;100(1):57-70).
• the Bcl-2 protein family comprises key regulators of cell survival which can suppress (e.g., Bcl-2, Bcl-xL, Mcl-1) or promote (e.g., Bad, Bax) apoptosis (Gross et al., Genes Dev. 1999 Aug 1;13(15):1899-911, Youle and Strasser, Nat. Rev. Mol. Cell Biol.2008 Jan;9(1):47- 59). In the face of stress stimuli, whether a cell survives or undergoes apoptosis is dependent on the extent of pairing between the Bcl-2 family members that promote cell death with family members that promote cell survival.
• Bcl-2 homology 3 (BH3) domain of proapoptotic family members into a groove on the surface of pro-survival members.
• Bcl-2 homology 3 (BH) domain defines the membership of the Bcl-2 family, which is divided into three main groups depending upon the particular BH domains present within the protein.
• the prosurvival members such as Bcl-2, Bcl- xL, and Mcl-1 contain BH domains 1–4, whereas Bax and Bak, the proapoptotic effectors of mitochondrial outer membrane permeabilization during apoptosis, contain BH domains 1–3 (Youle and Strasser, Nat. Rev. Mol.
• Bcl-xL (also named BCL2L1, from BCL2-like 1) is frequently amplified in cancer (Beroukhim et al., Nature 2010 Feb 18;463(7283):899-905) and it has been shown that its expression inversely correlates with sensitivity to more than 120 anti- cancer therapeutic molecules in a representative panel of cancer cell lines (NCI-60) (Amundson et al., Cancer Res.2000 Nov 1;60(21):6101-10).
• T-cells isolated from the joints of rheumatoid arthritis patients exhibited increased Bcl-xL expression and were resistant to spontaneous apoptosis (Salmon et al., J Clin Invest. 1997 Feb 1;99(3):439-46. doi: 10.1172/JCI119178.PMID: 9022077).
• the use of BH3 mimetics has also shown benefit in pre- clinical models of diseases of the immune system and autoimmune diseases. Treatment with ABT-737 (Bcl-2, Bcl-xL, and Bcl-w inhibitor) resulted in potent inhibition of lymphocyte proliferation in vitro.
• mice treated with ABT-737 in animal models of arthritis and lupus showed a significant decrease in disease severity (Bardwell et al., J Clin Invest. 1997 Feb 1;99(3):439-46. doi: 10.1172/JCI119178.PMID: 9022077).
• ABT ⁇ 737 prevented allogeneic T ⁇ cell activation, proliferation, and cytotoxicity in vitro and inhibited allogeneic T ⁇ and B ⁇ cell responses after skin transplantation with high selectivity for lymphoid cells (Cippa et al., .Transpl Int. 2011 Jul;24(7):722-32. doi: 10.1111/j.1432- 2277.2011.01272.x.
• BH3 mimetics a new class of drugs named BH3 mimetics. These molecules are able to disrupt the interaction between the proapoptotic and anti-apoptotic members of the Bcl-2 family and are potent inducers of apoptosis.
• This new class of drugs includes inhibitors of Bcl-2, Bcl-xL, Bcl-w and Mcl-1.
• the first BH3 mimetics described were ABT-737 and ABT-263, targeting Bcl-2, Bcl-xL and Bcl-w (Park et al., J. Med.
• ABT-263 has shown activity in several hematological malignancies and solid tumors (Shoemaker et al., Clin. Cancer Res.2008 Jun 1;14(11):3268-77; Ackler et al., Cancer Chemother. Pharmacol.2010 Oct;66(5):869-80; Chen et al., Mol. Cancer Ther.2011 Dec;10(12):2340-9).
• ABT-263 exhibited objective antitumor activity in lymphoid malignancies (Wilson et al., Lancet Oncol.2010 Dec;11(12):1149-59; Roberts et al., J. Clin. Oncol.
• Mcl-1 selective inhibitors have shown promising in vivo activity in several types of hematological cell malignancies in preclinical models and three of them, S64315, AMG176 and AZD5991, are currently being investigated in clinical trials (Yang et al., Eur. J. Med. Chem.2019 May 8;177:63-75). Therefore, BH3 mimetics represent a highly attractive approach for the development of novel therapies in oncology and in the field of immune and autoimmune diseases.
• BH3 mimetics and more specifically Bcl-xL inhibitors due to the medical need, can be upgraded by converting their mechanism of action from inhibition to degradation via PROTAC (Proteolysis-targeting chimeras).
• degraders are small molecules including (i) a ligand targeting a protein of interest (BH3) to be degraded, (ii) an E3 ubiquitin ligase recruitment ligand (mainly Cereblon (CRBN) or Von Hippel-Lindau (VHL) ligands), and (iii) a chemical linker connecting the two ligands.
• BH3 mimetics have been converted into PROTACs to date, such as Bcl-xL/Bcl- 2 dual inhibitors (DT2216 and XZ-739 based on ABT-263 scaffold, Khan et al., Nat Med 2019 Dec;25(12):1938-1947; Zhang et al., Eur. J. of Med. Chem.2020 Apr 15;192:112186), Bcl-xL inhibitors (XZ-424 based on A-1155463 scaffold, Zhang et al., Chem. Commun.2019 Dec 5; 55(98):14765-14768), Mcl-1 inhibitors (dMCL1-2 based on A-1210477 scaffold, Papatzimas et al., J.
• potent PROTACs that selectively degrade the Bcl-xL protein.
• the present disclosure fulfills this need.
• the present disclosure more specifically relates, in some embodiments, to pro-apoptotic agents that are capable of binding and killing cancer cells.
• the pro-apoptotic agents are PROTAC compounds comprising a linker that attaches a Bcl-xL inhibitor to an E3 ubiquitin ligase recruitment ligand.
• PROTAC compounds of the present disclosure can be represented by Formula (A): D ⁇ L ⁇ DSM (A), or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein: DSM is a degradation signaling compound covalently attached to the linker L; L is a linker that covalently attaches DSM to D; and D is a Bcl-xL inhibitor compound of Formula (I) or Formula (II) covalently attached to the linker L: or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein: ⁇ R 1 and R 2 independently of one another represent a group selected from the group consisting of: hydrogen; a linear or branched C 1 -C 6 alkyl optionally substituted by a hydroxyl or a C 1 -C 6 alkoxy group;
• PROTAC compounds of the present disclosure can be represented by Formula (A): D ⁇ L ⁇ DSM (A), or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein: DSM is a degradation signaling compound (e.g., an E3 ubiquitin ligase recruitment ligand, such as a CRBN ligand or a VHL ligand) covalently attached to a linker L; L is a linker that covalently attaches DSM to D; and D is a Bcl-xL inhibitor compound of Formula (I) or Formula (II) covalently attached to the linker L: or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein: ⁇ R 1 and R 2 independently of one another represent a group selected from: hydrogen; linear or branchedidididisomer, and/
• the linker L comprises at least one group selected from the group consisting of: a linear or branched C 1 -C 20 alkylene optionally substituted by one to three groups selected from the group consisting of a C 1 -C 8 alkyl, a C 3 -C 8 cycloalkyl, trifluoromethyl, hydroxyl, a halogen, and a C 1 - C 6 alkoxy; a C 3 -C 10 cycloalkylene; a C 3 -C 8 heterocycloalkylene; ⁇ C(O) ⁇ ; ⁇ O ⁇ ; ⁇ S ⁇ ; ⁇ N(R 16 ) ⁇ ; ⁇ N(R 16 )-C(O) ⁇ ; ⁇ C(O)-N(R 16 ) ⁇ ; ⁇ CH 2 ⁇ C(O) ⁇ N(
• the linker L of formula (A) comprises at least one group selected from: a linear or branched C 1 - C 20 alkylene optionally substituted by one or two groups selected from C 1 -C 8 alkyl, C 3 - C 8 cycloalkyl, trifluoromethyl, hydroxyl, halogen, and C 1 -C 6 alkoxy; a C 3 -C 10 cycloalkylene; ⁇ C(O) ⁇ ; ⁇ O ⁇ ; ⁇ S ⁇ ; ⁇ N(R 16 ) ⁇ ; ⁇ N(R 16 )-C(O) ⁇ ; ⁇ C(O)-N(R 16 ) ⁇ ; ⁇ CH 2 ⁇ C(O) ⁇ N(R 16 ) ⁇ ; ⁇ N(R 16 ) ⁇ C(O) ⁇ CH 2 ⁇ ; a poly
• the linker L of formula (A) comprises a 1,2,3-triazolene group formed by reacting an azide- containing precursor with an alkyne-containing precursor; and the remaining variables are as described in the first, second, third or fourth embodiment.
• LK 1 is a bond, ⁇ NR 16 ⁇ or ⁇ C(O) ⁇
• LK 2 is a bond, ⁇ C(O) ⁇ or ⁇ N(R 16 ) ⁇ C(O) ⁇ CH 2 ⁇ *
• LK 3 is ⁇ C(O) ⁇ or ⁇ N(R 16 ) ⁇ C(O) ⁇ CH 2 ⁇ *
• LK 4 is a bond or ⁇ C(O) ⁇
• LK 5 is a bond or ⁇ C(O) ⁇
• LK 6 is a bond, ⁇ C(O) ⁇ , ⁇ O ⁇ CH 2 ⁇ C(O) ⁇ *, or ⁇ N(R 16 ) ⁇ C(
• the linker L of formula (A) is represented by formula (i), (ii), (iii), (iv), (v), or (vi): wherein: LK 1 is a bond or ⁇ C(O) ⁇ ; LK 2 is a bond, ⁇ C(O) ⁇ or ⁇ N(R 16 ) ⁇ C(O) ⁇ CH 2 ⁇ *; LK 3 is ⁇ C(O) ⁇ or ⁇ N(R 16 ) ⁇ C(O) ⁇ CH 2 ⁇ *; LK 4 is a bond or ⁇ C(O) ⁇ ; LK 5 is a bond or ⁇ C(O) ⁇ ; LK 6 is a bond, ⁇ O ⁇ CH 2 ⁇ C(O) ⁇ *, or ⁇ N(R 16 ) ⁇ C(O) ⁇ CH 2 ⁇ *; R 16 is H or
• the linker L of formula (A) is selected from (L1)-(L50).
• D comprises a compound of Formula (I): , or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein: ⁇ R 1 and R 2 independently of one another represent a group selected from the group consisting of: hydrogen; a linear or branched C 1 -C 6 alkyl optionally substituted by a hydroxyl or a C 1 -C 6 alkoxy group; a C 3 -C 6 cycloalkyl; trifluoromethyl; and a linear or branched C 1 -C 6 alkylene-heterocycloalkyl wherein the heterocycloalkyl group is optionally substituted by a linear or branched C 1 -C 6 alkyl group; or R 1 and R 2 form with
• D in Formula (A) comprises a compound of Formula (I): , wherein: ⁇ R 1 and R 2 independently of one another represent a group selected from: hydrogen; linear or branched C 1 -C 6 alkyl optionally substituted by a hydroxyl or a C 1 -C 6 alkoxy group; C 3 -C 6 cycloalkyl; trifluoromethyl; linear or branched C 1 -C 6 alkylene-heterocycloalkyl wherein the heterocycloalkyl group is optionally substituted by a a linear or branched C 1 -C 6 alkyl group; or R 1 and R 2 form with the carbon atoms carrying them a C 3 -C 6 cycloalkylene group, ⁇ R 3 represents a group selected from: hydrogen; C 3 branched C 1 -C 6 alkyl optionally substituted by a hydroxyl or a C 1 -C 6 alkoxy group; C 3 -C 6 cycloalkyl; triflu
• R 1 is linear or branched C 1 -C 6 alkyl and R 2 is H; and the remaining variables are as described in the ninth or tenth embodiment.
• R G4 is selected from the group consisting of hydrogen, a C 1 -C 6 alkyl optionally substituted by 1 to 3 halogen atoms, a C 1 -C 6 alkyl substituted by a hydroxyl, a C 1 -C 6 alkyl substituted by a C 1 -C 6 alkoxy group, a C 2 -C 6 alkenyl, a C 2 -C 6 alkynyl and a C 3 -C 6 cycloalkyl, and R G5 represents a hydrogen atom or a C 1 -C 6 alkyl group optionally substituted by 1 to 3 halogen atoms, ⁇ R 4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl, a hydroxyl or a methoxy group, ⁇ R 5 represents a group selected from the group consisting of: a C 1 -C 6 alkyl optionally substituted by 1 to 3 halogen atoms; a C 2 -C
• ⁇ A 4 and A 5 independently of one another represent a carbon or a nitrogen atom
• ⁇ Z 1 represents a bond, -N(R)-, or –O-, wherein R represents a hydrogen or a linear or branched C 1 -C 6 alkyl
• ⁇ R 1 represents a group selected from: hydrogen; linear or branched C 1 -C 6 alkyl optionally substituted by a hydroxyl or a C 1 -C 6 alkoxy group; C 3 -C 6 cycloalkyl; trifluoromethyl; linear or branched C 1 -C 6 alkylene-heterocycloalkyl wherein the heterocycloalkyl group is optionally substituted by a a linear or branched C 1 -C 6 alkyl group;
• ⁇ R 2 represents a hydrogen or a methyl;
• ⁇ R 3 represents a group selected from: hydrogen; linear or branched C 1 -C 4 alkyl; -X 1 -
• a 4 and A 5 both represent a nitrogen atom
• R 1 is linear or branched C 1- 6 alkyl
• R 2 is H
• n is 1
• the remaining variables are as described in the twelfth or thirteenth embodiment.
• D comprises a compound of formula (IA) or (IIA): or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein: ⁇ Z 1 represents a bond or –O-, ⁇ R 3 represents a group selected from the group consisting of: hydrogen; a C 3 - C 6 cycloalkyl; a linear or branched C 1 -C 6 alkyl; -X 1 -NR a R b ; -X 1 -N + R a R b R c ; -X 1 -O-R c ; - X 1 -N 3 and ⁇ R a and R b independently of one another represent a group selected from the group consisting of: hydrogen;
• D comprises a compound of formula (IA) or (IIA): wherein: ⁇ Z 1 represents a bond or –O-, ⁇ R 3 represents a group selected from: hydrogen; C 3 -C 6 cycloalkyl; linear or branched C 1 -C 6 alkyl; -X 1 -NR a R b ; -X 1 -N + R a R b R c ; -X 1 -O-R c ; -X 1 -N 3 and ⁇ R a and R b independently of one another represent a group selected from: hydrogen; linear or branched C 1 -C 6 alkyl optionally substituted by one or two hydroxyl groups; and C 1 -C 6 alkylene-SO 2 O-, ⁇ R c represents a hydrogen
• R 7 represents a group selected from: linear or branched C 1 -C 6 alkyl group; (C 3 - C 6 )cycloalkylene-R 8 ; or: wherein: Cy represents a C 3 -C 8 cycloalkyl; and the remaining variables are as described in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth or sixteenth embodiment.
• R 7 represents a group selected from: and the remaining variables are as described in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth or seventeenth embodiment.
• R 3 represents a group selected from the group consisting of: hydrogen; a linear or branched C 1 -C 6 alkyl; -X 1 -NR a R b ; -X 1 -N + R a R b R c ; -X 1 -O-R c ; - X 1 -N 3 and ⁇ for formula (IIB) or (IIC-1),
• Z 1 represents a bond
• R 3 represents hydrogen; or Z 1 represents –O-, and R 3 represents –X 1 -NR a R b ,
• G is selected from the group consisting of –C(O)OH and - C(O)NR G1 R G2 ; ⁇ for formula (IIC-1), G is selected from the group consisting of –
• D in Formula (A) comprises a compound of formula (IB), (IC), (IIB) or (IIC):
• R 3 represents a group selected from: hydrogen; linear or branched C 1 -C 6 alkyl ; -X 1 -NR a R b ; -X 1 -N + R a R b R c ; -X 1 -O-R c ; -X 1 -N 3 and for formula (IIB) or (IIC), Z1 represents a bond, and R 3 represents hydrogen; or Z 1 represents –O-, and R 3 represents –X 1 -NR a R b , ⁇ R a and R b independently of one another represent a group selected from: hydrogen; linear or branched C 1 -C 6 alkyl optionally substituted by one or two hydroxyl groups; and C 1 -C 6 alkylene-SO 2 O-, ⁇ R c represents a hydrogen or a linear or branched C 1 -C 6 alkyl group ⁇ R 6 represents a linear or branched –
• R6 represents ⁇ X 2 -O-R 7
• R 7 represents the following group: and the remaining variables are as described in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth or twentieth embodiment.
• R 6 represents a heteroarylene-R 7 group optionally substituted by a linear or branched C 1 -C 6 alkyl group
• R 7 represents a group selected from: and the remaining variables are as described in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth or twentieth embodiment.
• B 3 represents a C 3 -C 8 heterocycloalkyl group selected from a pyrrolidinyl group, a piperidinyl group, a piperazinyl group, a morpholinyl group, an azepanyl group, and a 4,4- difluoropiperidin-1-yl group; and the remaining variables are as described in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth, twentieth, twenty-first or twenty- second embodiment.
• B 3 represents a pyrrolidinyl group or a piperazinyl group and the remaining variables are as described in the twenty-third embodiment.
• B 3 represents a piperazinyl group; and the remaining variables are as described in the twenty- third embodiment.
• R 8 represents a group selected from the group consisting of: , wherein: is a bond to the linker; and the remaining variables are as described in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth, twentieth, twenty-first or twenty-second embodiment.
• R 8 represents a group selected from: wherein: is a bond to the linker; and the remaining variables are as described in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth, twentieth, twenty-first or twenty-second embodiment.
• R 3 represents a group selected from the group consisting of: hydrogen; a linear or branched C 1 -C 6 alkyl; -X 1 -NR a R b ; -X 1 -N 3 and ⁇ for formula (IIB) or (IIC-1), Z 1 represents a bond, and R 3 represents hydrogen, ⁇ for formula (IC-1), G is selected from the group consisting of –C(O)OH and - C(O)N(CH 3 ) 2 ; ⁇ for formula (IIC-1), G is selected from the group consisting of -C(O)NHS(O) 2 H, -C(O)NH 2 , -C(O)NHCH 3 , -C(O)NHC(CH 3 , -C(O)NHC(CH 3 ),
• the variables are defined as: ⁇ for formula (IB) or (IC), R 3 represents a group selected from: hydrogen; linear or branched C 1 -C 6 alkyl; -X 1 -N 3 and , for formula (IIB) or (IIC), Z 1 represents a bond, and R 3 represents hydrogen, ⁇ R6 represents a linear or branched –C 1 -C 6 alkylene-R 8 group, –X 2 -O-R 7 or a heteroarylene-R 7 group optionally substituted by a linear or branched C 1 -C 6 alkyl group, ⁇ R 7 represents a group selected from: ⁇ R 8 represents a group selected from: -NR’ a R’ b ; and -
• B 3 represents pyrrolidinyl group or a piperazinyl group and the remaining variables are as described in the twenty-eighth or twenty-ninth embodiment
• B 3 represents a piperazinyl group; and the remaining variables are as described in the twenty-eighth or twenty-ninth embodiment.
• R 8 represents a group selected from the group consisting of: , wherein: is a bond to the linker; and the remaining variables are as described in the twenty-eighth or twenty-ninth embodiment.
• R 8 represents a group selected from: w herein: is a bond to the linker; and the remaining variables are as described in the twenty-eighth or twenty-ninth embodiment.
• D represents any one of the following attached to L:
• D ⁇ L in Formula (A) comprises a formula selected from:
• DSM in Formula (A) is a E3 ligase recognition agent; and the remaining variables are as described in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth, twentieth, twenty-first, twenty-second, twenty-third, twenty-fourth, twenty-fifth, twenty-sixth, twenty-seventh, twenty-eighth, twenty-ninth, thirtieth, thirty-first, thirty-second, thirty-third, thirty-fourth or thirty-fifth embodiment.
• DSM in Formula (A) is a VHL ligand, a thalidomide cereblon binder or an inhibitor of apoptosis (IAP) E3 ligases; and the remaining variables are as described in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth, twentieth, twenty-first, twenty- second, twenty-third, twenty-fourth, twenty-fifth, twenty-sixth, twenty-seventh, twenty-eighth, twenty-ninth, thirtieth, thirty-first, thirty-second, thirty-third, thirty-fourth or thirty-fifth embodiment.
• DSM in Formula (A) represents any one of the following attached to L: or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein: represents a bond to the linker (L); and the remaining variables are as described in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth, twentieth, twenty-first, twenty-second, twenty-third, twenty-fourth, twenty-fifth, twenty-sixth, twenty-seventh, twenty-eighth, twenty-ninth, thirtieth, thirty-first, thirty-second, thirty-third, thirty-fourth or thirty-fif
• DSM represents the following attached to L: or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein: represents a bond to the linker (L); and the remaining variables are as described in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth, twentieth, twenty-first, twenty-second, twenty-third, twenty-fourth, twenty-fifth, twenty-sixth, twenty-seventh, twenty-eighth, twenty-ninth, thirtieth, thirty-first, thirty-second, thirty-third, thirty-fourth or thirty-fifth embodiment.
• the compound of the present disclosure is any one of the compounds in Table 7, or an enantiomer, a diastereomer, and/or a pharmaceutically acceptable salt thereof.
• the present disclosure also provides pharmaceutical compositions comprising a PROTAC compounds describe herein (e.g., the compound of the first to twenty-eighth embodiments described above) and a pharmaceutically acceptable carrier.
• the present disclosure also relates to a method of treating a subject having or suspected of having a cancer comprises administering to the subject a therapeutically effective amount of a compound described herein (e.g., the compound of the first to twenty- eighth embodiments described above) or a pharmaceutical composition thereof.
• the cancer is a solid tumor or a hematological cancer.
• the cancer is a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular cancer, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or spleen cancer.
• the PROTAC compound is administered as monotherapy. [059] In some embodiments, the PROTAC compound is administered adjunctive to another therapeutic agent or radiation therapy. [060] In some embodiments, the PROTAC compound is administered in an amount effective to sensitize the tumor cells to one or more additional therapeutic agents and/or radiation therapy. [061] In some embodiments, the methods described above further comprise administering to the subject in need thereof at least one additional therapeutic agent. [062] In some embodiments, the additional therapeutic agent is a Bcl-2 inhibitor, a taxane, a MEK inhibitor, an ERK inhibitor, or a RAF inhibitor.
• PROTAC compound of Formula (A) for use in a method described above (e.g., a method of treating a subject having or suspected of having a cancer).
• the present disclosure also relates to the use of a PROTAC compound of Formula (A) for the manufacture of a medicament for treating a subject having or suspected of having a cancer.
• DETAILED DESCRIPTION [064]
• the disclosed compositions and methods may be understood more readily by reference to the following detailed description . [065] Throughout this text, the descriptions refer to compositions and methods of using the compositions. Where the disclosure describes or claims a feature or embodiment associated with a composition, such a feature or embodiment is equally applicable to the methods of using the composition.
• compositions and methods which are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosed compositions and methods that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any sub-combination.
• PROTAC compounds can be identified using a naming convention in the general format of “DSM ⁇ linker ⁇ Bcl-xL inhibitor compound.” For example only, if a compound is referred to as “DSM1a-L1-D1a”, such a compound would comprise a DSM designated as DSM1a, a linker designated as L1, and a Bcl-xL inhibitor compound moiety designated as D1a. Similar designation can be used to identify components or moieties in the PROTAC compounds described herein. [070] Any formula given herein is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds.
• lsotopically labeled compounds have structures depicted by the formulae given herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number.
• Isotopes that can be incorporated into compounds of this disclosure include, for example, isotopes of hydrogen, carbon, nitrogen, oxygen fluorine and chlorine such as 3 H, 11 C, 13 C, 14 C, 15 N, 18 F and 36 Cl. Accordingly it should be understood that the present disclosure includes compounds that incorporate one or more of any of the aforementioned isotopes, including for example, radioactive isotopes, such as 3 H and 14 C, or those into which non-radioactive isotopes, such as 2 H and 13 C are present.
• Such isotopically labelled compounds are useful in metabolic studies (with 14 C), reaction kinetic studies (with, for example 2 H or 3 H), detection or imaging techniques, such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT) including drug or substrate tissue distribution assays, or in radioactive treatment of patients.
• PET positron emission tomography
• SPECT single-photon emission computed tomography
• an 18F or labeled compound may be particularly desirable for PET or SPECT studies.
• Isotopically-labeled compounds can generally be prepared by conventional techniques known to those skilled in the art, e.g., using an appropriate isotopically-labeled reagents in place of the non-labeled reagent previously employed.
• alkyl refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation.
• C 1 -C 6 alkyl refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to six carbon atoms, and which is attached to the rest of the molecule by a single bond.
• Non- limiting examples of "C 1 -C 6 alkyl” groups include methyl (a C 1 alkyl), ethyl (a C 2 alkyl), 1- methylethyl (a C 3 alkyl), n-propyl (a C 3 alkyl), isopropyl (a C 3 alkyl), n-butyl (a C 4 alkyl), isobutyl (a C 4 alkyl), sec-butyl (a C 4 alkyl), tert-butyl (a C 4 alkyl), n-pentyl (a C 5 alkyl), isopentyl (a C 5 alkyl), neopentyl (a C 5 alkyl) and hexyl (a C 6 alkyl).
• alkenyl refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond.
• C 2 -C 6 alkenyl refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond, having from two to six carbon atoms, which is attached to the rest of the molecule by a single bond.
• C 2 -C 6 alkenyl groups include ethenyl (a C 2 alkenyl), prop-1-enyl (a C 3 alkenyl), but-1-enyl (a C 4 alkenyl), pent-1-enyl (a C 5 alkenyl), pent- 4-enyl (a C 5 alkenyl), penta-1,4-dienyl (a C 5 alkenyl), hexa-1-enyl (a C 6 alkenyl), hexa-2-enyl (a C 6 alkenyl), hexa-3-enyl (a C 6 alkenyl), hexa-1-,4-dienyl (a C 6 alkenyl), hexa-1-,5-dienyl (a C 6 alkenyl) and hexa-2-,4-dienyl (a C 6 alkenyl).
• C 2 -C 3 alkenyl refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond, having from two to three carbon atoms, which is attached to the rest of the molecule by a single bond.
• Non-limiting examples of "C 2 - C 3 alkenyl” groups include ethenyl (a C 2 alkenyl) and prop-1-enyl (a C 3 alkenyl).
• alkylene refers to a bivalent straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms and containing no unsaturation.
• C 1 -C 6 alkylene refers to a bivalent straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to six carbon atoms.
• Non-limiting examples of "C 1 -C 6 alkylene” groups include methylene (a C 1 alkylene), ethylene (a C 2 alkylene), 1- methylethylene (a C 3 alkylene), n-propylene (a C 3 alkylene), isopropylene (a C 3 alkylene), n- butylene (a C 4 alkylene), isobutylene (a C 4 alkylene), sec-butylene (a C 4 alkylene), tert-butylene (a C 4 alkylene), n-pentylene (a C 5 alkylene), isopentylene (a C 5 alkylene), neopentylene (a C 5 alkylene), and hexylene (a C 6 alkylene).
• alkenylene refers to a bivalent straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms and containing at least one double bond.
• C 2 -C 6 alkenylene refers to a bivalent straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond, and having from two to six carbon atoms.
• Non- limiting examples of "C 2 -C 6 alkenylene” groups include ethenylene (a C 2 alkenylene), prop-1- enylene (a C 3 alkenylene), but-1-enylene (a C 4 alkenylene), pent-1-enylene (a C 5 alkenylene), pent-4-enylene (a C 5 alkenylene), penta-1,4-dienylene (a C 5 alkenylene), hexa-1-enylene (a C 6 alkenylene), hexa-2-enylene (a C 6 alkenylene), hexa-3-enylene (a C 6 alkenylene), hexa-1- ,4-dienylene (a C 6 alkenylene), hexa-1-,5-dienylene (a C 6 alkenylene) and hexa-2-,4-dienylene (a C 6 alkenylene)
• C 2 -C 3 alkenylene groups include ethenylene (a C 2 alkenylene) and prop-1- enylene (a C 3 alkenylene).
• aryl refers to a phenyl, naphthyl, biphenyl or indenyl group.
• cycloalkyl refers to any mono- or bi-cyclic non-aromatic carbocyclic group containing from 3 to 10 ring members, which may include fused, bridged or spiro ring systems.
• Non-limiting examples of fused bicyclic or bridged polycyclic ring systems include bicyclo[1.1.1]pentane, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, bicyclo[3.1.1] heptane, bicyclo[3.2.1]octane, bicyclo[2.2.2]octane and adamantanyl.
• Non-limiting examples monocyclic C 3 -C 8 cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl groups.
• cycloalkylene refers to a cycloalkyl, as defined herein, having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent cycloalkyl.
• examples of cycloalkylene include, but are not limited to, cyclopropylene, cyclobutylene, cyclopentylene and cyclohexylene.
• Cycloalkylenes of the present disclosure include monocyclic, bicylic and tricyclic ring structures.
• haloalkyl refers to a linear or branched alkyl chain substituted with one or more halogen groups in place of hydrogens along the hydrocarbon chain.
• halogen groups suitable for substitution in the haloalkyl group include Fluorine, Bromine, Chlorine, and Iodine.
• Haloalkyl groups may include substitution with multiple halogen groups in place of hydrogens in an alkyl chain, wherein said halogen groups can be attached to the same carbon or to another carbon in the alkyl chain.
• heteroaryl refers any mono- or bi-cyclic group composed of from 5 to 10 ring members, having at least one aromatic moiety and containing from 1 to 4 hetero atoms selected from oxygen, sulphur and nitrogen (including quaternary nitrogens).
• heterocycloalkyl means any mono- or bi-cyclic non-aromatic carbocyclic group, composed of from 3 to 10 ring members, and containing from one to 3 hetero atoms selected from oxygen, sulphur, SO, SO 2 and nitrogen, it being understood that bicyclic group may be fused or spiro type.
• C 3 -C 8 heterocycloalkyl refers to heterocycloalkyl having 3 to 8 ring carbon atoms.
• the heterocycloalkyl can have 4 to 10 ring members.
• heteroarylene and heterocycloalkylene mean divalent heteroaryl and heterocycloalkyl groups, including heterocyclic groups with bicylic and tricyclic ring structures.
• the alkyl, alkenyl, alkynyl, alkoxy, amino, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl groups may be optionally substituted by 1 to 4 groups selected from optionally substituted linear or branched (C 1 -C 6 )alkyl, optionally substituted linear or branched (C 2 -C 6 )alkenyl group, optionally substituted linear or branched (C 2 -C 6 )alkynyl group, optionally substituted linear or branched (C 1 -C 6 )alkoxy, optionally substituted (C 1 -C 6 )alkyl-S-, hydroxy, oxo (or N-oxide where appropriate), nitro, cyano, -C(O)-OR 0 ’, -O-C(O)-R 0 ’, -C(O)-NR 0 ’R 0 ”, - NR 0 ,
• linker refers to a chemical moiety in Formula (A) that connects D to DSM.
• polyoxyethylene refers to a linear chain, a branched chain or a star shaped configuration comprised of (OCH 2 CH 2 ) groups.
• polyalkylene glycol refers to a linear chain, a branched chain or a star shaped configuration comprised of (O(CH 2 ) m ) n groups.
• about means plus or minus 20%, 15%, 10%, 5%, 1%, 0.5%, or 0.1% of a numerical amount.
• the term “about” refers to a range of values which are 10% more or less than the specified value.
• the term “about” refers to a range of values which are 5% more or less than the specified value.
• the term “about” refers to a range of values which are 1% more or less than the specified value.
• the term “agent” is used herein to refer to a chemical compound, a mixture of chemical compounds, a biological macromolecule, an extract made from biological materials, or a combination of two or more thereof.
• chemotherapeutic agent refers to an agent that is capable of modulating a biological process and/or has biological activity.
• the Bcl-xL inhibitors and the PROTAC compounds comprising them, as described herein, are exemplary therapeutic agents.
• chemotherapeutic agent or “anti-cancer agent” is used herein to refer to all agents that are effective in treating cancer (regardless of mechanism of action). Inhibition of metastasis or angiogenesis is frequently a property of a chemotherapeutic agent.
• Chemotherapeutic agents include antibodies, biological molecules, and small molecules, and encompass the Bcl-xL inhibitors and DSM conjugates comprising them, as described herein.
• a chemotherapeutic agent may be a cytotoxic or cytostatic agent.
• cytostatic agent refers to an agent that inhibits or suppresses cell growth and/or multiplication of cells.
• cytotoxic agent refers to a substance that causes cell death primarily by interfering with a cell’s expression activity and/or functioning.
• PROTAC conjugate refers to one or more therapeutic compounds (e.g., a Bcl-xL inhibitor) that is covalently linked to a DSM such as an E3 ubiquitin ligase recruitment ligand.
• a Bcl-xL inhibitor e.g., a Bcl-xL inhibitor
• DSM degradation signaling moiety
• L a linker moiety
• D a drug moiety (e.g., a Bcl-xL inhibitor drug moiety).
• DSM degradation signaling moiety
• DSMs of this disclosure degrade targeted proteins by binding or recruiting at least one degradation protein, which is usually associated with the proteasome, the ubiquitin-proteasome pathways, or lysosomal proteolysis.
• DSMs of this disclosure include, but are not limited to, E3 ligase recognition or recruitment ligand.
• ubiquitin ligase refers to a family of proteins that facilitate the transfer of ubiquitin to a specific substrate protein, targeting the substrate protein for degradation. Cereblon, for example, is an E3 Ubiquitin Ligase protein that alone or in combination with an E2 ubiqutin-conjugating enzyme causes the attachment of ubiquitin to a lysine on a target protein, and subsequently targets the specific protein substrate for degradation by the proteasome.
• B-cell lymphoma-extra large refers to any native form of human Bcl-xL, an anti-apoptotic member of the Bcl-2 protein family.
• the term encompasses full-length human Bcl-xL (e.g., UniProt Reference Sequence: Q07817-1; SEQ ID NO:71), as well as any form of human Bcl-xL that may result from cellular processing.
• the term also encompasses functional variants or fragments of human Bcl-xL, including but not limited to splice variants, allelic variants, and isoforms that retain one or more biologic functions of human Bcl-xL (i.e., variants and fragments are encompassed unless the context indicates that the term is used to refer to the wild-type protein only).
• Bcl-xL can be isolated from human, or may be produced recombinantly or by synthetic methods.
• the term “inhibit” or “inhibition” or “inhibiting,” as used herein, means to reduce a biological activity or process by a measurable amount, and can include but does not require complete prevention or inhibition.
• “inhibition” means to reduce the expression and/or activity of Bcl-xL and/or one or more upstream modulators or downstream targets thereof.
• Bcl-xL inhibitor refers to an agent capable of reducing the expression and/or activity of Bcl-xL and/or one or more upstream modulators or downstream targets thereof.
• Exemplary Bcl-xL modulators are described in WO2010/080503, WO2010/080478, WO2013/055897, WO2013/055895, WO2016/094509, WO2016/094517, WO2016/094505, Tao et al., ACS Medicinal Chemistry Letters (2014), 5(10), 1088-109, and Wang et al., ACS Medicinal Chemistry Letters (2020), 11(10), 1829 ⁇ 1836, each of which are incorporated herein by reference as exemplary Bcl-xL modulators, including exemplary Bcl-xL inhibitors, that can be included as drug moieties in the PROTAC compounds described herein.
• a “Bcl-xL inhibitor drug moiety”, “Bcl-xL inhibitor moiety”, and the like refer to the component of the PROTAC compounds described herein that provides the structure of a Bcl-xL inhibitor compound or a compound modified for attachment to a DSM that retains essentially the same, similar, or enhanced biological function or activity as compared to the original compound.
• Bcl-xL inhibitor drug moiety is component (D) in a compound of Formula (A).
• cancer refers to the presence of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and/or certain morphological features.
• cancer cells can be in the form of a tumor or mass, but such cells may exist alone within a subject, or may circulate in the blood stream as independent cells, such as leukemic or lymphoma cells.
• the term "cancer” includes all types of cancers and cancer metastases, including hematological cancers, solid tumors, sarcomas, carcinomas and other solid and non- solid tumor cancers.
• Hematological cancers may include B-cell malignancies, cancers of the blood (leukemias), cancers of plasma cells (myelomas, e.g., multiple myeloma), or cancers of the lymph nodes (lymphomas).
• Exemplary B-cell malignancies include chronic lymphocytic leukemia (CLL), follicular lymphoma, mantle cell lymphoma, and diffuse large B-cell lymphoma.
• Leukemias may include acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myelomonocytic leukemia (CMML), acute monocytic leukemia (AMoL), etc.
• Lymphomas may include Hodgkin's lymphoma, non-Hodgkin's lymphoma, etc.
• Other hematologic cancers may include myelodysplasia syndrome (MDS).
• Solid tumors may include carcinomas such as adenocarcinoma, e.g., breast cancer, pancreatic cancer, prostate cancer, colon or colorectal cancer, lung cancer, gastric cancer, cervical cancer, endometrial cancer, ovarian cancer, cholangiocarcinoma, glioma, melanoma, etc.
• carcinomas such as adenocarcinoma, e.g., breast cancer, pancreatic cancer, prostate cancer, colon or colorectal cancer, lung cancer, gastric cancer, cervical cancer, endometrial cancer, ovarian cancer, cholangiocarcinoma, glioma, melanoma, etc.
• the cancer is a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular cancer, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin melanoma myelogenous leukemia myeloma oral cancer ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or spleen cancer.
• the cancer is a lymphoma or gastric cancer.
• tumor refers to any mass of tissue that results from excessive cell growth or proliferation, either benign or malignant, including precancerous lesions.
• the tumor is a breast cancer, gastric cancer, bladder cancer, brain cancer, cervical cancer, colorectal cancer, esophageal cancer, hepatocellular cancer, melanoma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, or spleen cancer.
• the tumor is a gastric cancer.
• tumor cell and “cancer cell” may be used interchangeably herein and refer to individual cells or the total population of cells derived from a tumor or cancer, including both non-tumorigenic cells and cancer stem cells.
• tumor cell and “cancer cell” will be modified by the term “non-tumorigenic” when referring solely to those cells lacking the capacity to renew and differentiate to distinguish those cells from cancer stem cells.
• subject and “patient” are used interchangeably herein to refer to any human or non-human animal in need of treatment. Non-human animals include all vertebrates (e.g., mammals and non-mammals) such as any mammal.
• Non-limiting examples of mammals include humans, chimpanzees, apes, monkeys, cattle, horses, sheep, goats, swine, rabbits, dogs, cats, rats, mice, and guinea pigs.
• Non-limiting examples of non-mammals include birds and fish.
• the subject is a human.
• the term “a subject in need of treatment,” as used herein, refers to a subject that would benefit biologically, medically, or in quality of life from a treatment (e.g., a treatment with any one or more of the exemplary compounds described herein).
• treatment refers to any improvement of any consequence of disease, disorder, or condition, such as prolonged survival, less morbidity, and/or a lessening of side effects which result from an alternative therapeutic modality.
• treatment comprises delaying or ameliorating a disease, disorder, or condition (i.e., slowing or arresting or reducing the development of a disease or at least one of the clinical symptoms thereof).
• treatment comprises delaying, alleviating, or ameliorating at least one physical parameter of a disease, disorder, or condition, including those which may not be discernible by the patient.
• treatment comprises modulating a disease disorder or condition either physically (eg stabilization of a discernible symptom), physiologically (e.g., stabilization of a physical parameter), or both.
• treatment comprises administration of a described compound or composition to a subject, e.g., a patient, to obtain a treatment benefit enumerated herein.
• the treatment can be to cure, heal, alleviate, delay, prevent, relieve, alter, remedy, ameliorate, palliate, improve, or affect a disease, disorder, or condition (e.g., a cancer), the symptoms of a disease, disorder, or condition (e.g., a cancer), or a predisposition toward a disease, disorder, or condition (e.g., a cancer).
• a composition disclosed herein in addition to treating a subject having a disease, disorder, or condition, can also be provided prophylactically to prevent or reduce the likelihood of developing that disease, disorder, or condition.
• the term “prevent”, “preventing,” or “prevention” of a disease, disorder, or condition refers to the prophylactic treatment of the disease, disorder, or condition; or delaying the onset or progression of the disease, disorder, or condition.
• a "pharmaceutical composition” refers to a preparation of a composition, e.g., an compound or composition, in addition to at least one other (and optionally more than one other) component suitable for administration to a subject, such as a pharmaceutically acceptable carrier, stabilizer, diluent, dispersing agent, suspending agent, thickening agent, and/or excipient.
• a pharmaceutically acceptable carrier such as a pharmaceutically acceptable sulfate, a pharmaceutically acceptable carrier, stabilizer, diluent, dispersing agent, suspending agent, thickening agent, and/or excipient.
• the pharmaceutical compositions provided herein are in such form as to permit administration and subsequently provide the intended biological activity of the active ingredient(s) and/or to achieve a therapeutic effect.
• the pharmaceutical compositions provided herein preferably contain no additional components which are unacceptably toxic to a subject to which the formulation would be administered.
• Pharmaceutically acceptable carriers may enhance or stabilize the composition or can be used to facilitate preparation of the composition.
• Pharmaceutically acceptable carriers can include solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, drug stabilizers, binders, excipients, disintegration agents, lubricants, sweetening agents, flavoring agents, dyes, and the like and combinations thereof, as would be known to those skilled in the art (see, for example, Remington's Pharmaceutical Sciences 18th Ed Mack Printing Company 1990 pp 1289- 1329). Except insofar as any conventional carrier is incompatible with the active ingredient, its use in the therapeutic or pharmaceutical compositions is contemplated.
• preservatives e.g., antibacterial agents, antifungal agents
• isotonic agents e.g., absorption delaying agents, salts, preservatives, drug stabilizers, binders, excipients, disintegration agents, lubricants, sweetening agents, flavoring agents
• the carrier may be selected to minimize adverse side effects in the subject, and/or to minimize degradation of the active ingredient(s).
• An adjuvant may also be included in any of these formulations.
• the term "excipient” refers to an inert substance added to a pharmaceutical composition to further facilitate administration of an active ingredient.
• Formulations for parenteral administration can, for example, contain excipients such as sterile water or saline, polyalkylene glycols such as polyethylene glycol, vegetable oils, or hydrogenated napthalenes.
• excipients include, but are not limited to, calcium bicarbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, ethylene-vinyl acetate co-polymer particles, and surfactants, including, for example, polysorbate 20.
• pharmaceutically acceptable salt refers to a salt which does not abrogate the biological activity and properties of the compounds of the present disclosure, and does not cause significant irritation to a subject to which it is administered.
• salts include, but are not limited to: (a) acid addition salts formed with inorganic acids, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid and the like; and salts formed with organic acids, for example, acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic acid, and the like; and (b) salts formed from elemental anions such as chlorine, bromine, and iodine.
• inorganic acids for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phospho
• the PROTAC compounds, linkers, Bcl-xL inhibitors and linker-Bcl-xL inhibitors described herein can contain a monovalent anionic counterion M 1 -. Any suitable anionic counterion can be used.
• the monovalent anionic counterion is a pharmaceutically acceptable monovalent anionic counterion.
• the monovalent anionic counterion M 1 - can be selected from bromide, chloride, iodide, acetate, trifluoroacetate, benzoate, mesylate, tosylate, triflate, formate, or the like.
• the monovalent anionic counterion M 1 - is trifluoroacetate or formate
• the term “therapeutically effective amount” or “therapeutically effective dose,” refers to an amount of a compound described herein, e.g., a PROTAC compound or composition described herein, to effect the desired therapeutic result (i.e., reduction or inhibition of an enzyme or a protein activity, amelioration of symptoms, alleviation of symptoms or conditions, delay of disease progression, a reduction in tumor size, inhibition of tumor growth, prevention of metastasis).
• a therapeutically effective amount does not induce or cause undesirable side effects.
• a therapeutically effective amount induces or causes side effects but only those that are acceptable by a treating clinician in view of a patient’s condition.
• a therapeutically effective amount is effective for detectable killing, reduction, and/or inhibition of the growth or spread of cancer cells, the size or number of tumors, and/or other measure of the level, stage, progression and/or severity of a cancer.
• the term also applies to a dose that will induce a particular response in target cells, e.g., a reduction, slowing, or inhibition of cell growth.
• a therapeutically effective amount can be determined by first administering a low dose, and then incrementally increasing that dose until the desired effect is achieved.
• a therapeutically effective amount can also vary depending upon the intended application (in vitro or in vivo), or the subject and disease condition being treated, e.g., the weight and age of the subject, the severity of the disease condition, the manner of administration and the like, which can readily be determined by one of ordinary skill in the art.
• the specific amount may vary depending on, for example, the particular pharmaceutical composition, the subject and their age and existing health conditions or risk for health conditions, the dosing regimen to be followed, the severity of the disease, whether it is administered in combination with other agents, timing of administration, the tissue to which it is administered, and the physical delivery system in which it is carried.
• a therapeutically effective amount of a PROTAC compound may reduce the number of cancer cells, reduce tumor size, inhibit (e.g., slow or stop) tumor metastasis, inhibit (e.g., slow or stop) tumor growth, and/or relieve one or more symptoms.
• the term “prophylactically effective amount” or “prophylactically effective dose,” refers to an amount of a compound disclosed herein, e.g., a PROTAC compound or composition described herein, that is effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease the prophylactically effective amount will be less than the therapeutically effective amount.
• a prophylactically effective amount can prevent the onset of disease symptoms, including symptoms associated with a cancer.
• PROTAC COMPOUNDS [114]
• the PROTAC compounds of the present disclosure include those with anti-cancer activity.
• the PROTAC compounds include a degradation signaling moiety (DSM) conjugated (i.e., covalently attached by a linker) to a drug moiety (e.g., a Bcl-xL inhibitor), wherein the drug moiety when not conjugated to a DSM has a cytotoxic or cytostatic effect.
• DSM degradation signaling moiety
• the drug moiety when not conjugated to a DSM is capable of reducing the expression and/or activity of Bcl-xL and/or one or more upstream modulators or downstream targets thereof.
• the PROTAC compounds disclosed herein may provide potent anti-cancer agents.
• the PROTAC compound may provide improved activity, better cytotoxic specificity, and/or reduced off-target killing as compared to the drug moiety when administered alone.
• the components of the PROTAC compounds are selected to (i) retain one or more therapeutic properties exhibited by the DSM in isolation, (ii) maintain the specific binding properties of the DSM; (iii) allow delivery, e.g., intracellular delivery, of the drug moiety via stable attachment to the DSM; (iv) retain PROTAC compound stability as an intact compound until transport or delivery to a target site; (v) allow for the therapeutic effect, e.g., cytotoxic effect, of the drug moiety after cleavage or other release mechanism in the cellular environment; (vi) exhibit in vivo anti-cancer treatment efficacy comparable to or superior to that of the DSM and drug moieties in isolation; (vii) minimize off- target killing by the drug moiety; and/or (viii) exhibit desirable pharmacokinetic and pharmacodynamics properties, formulatability, and toxicologic/immunologic profiles.
• PROTAC compounds comprising a degradation signaling moiety (DSM), a Bcl-xL inhibitor drug moiety (D), and a linker moiety (L) that covalently attaches DSM to D.
• DSM degradation signaling moiety
• D Bcl-xL inhibitor drug moiety
• L linker moiety
• PROTAC compounds of the present disclosure have the Formula (A): D ⁇ L ⁇ DSM (A), or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the forgoing, wherein DSM is a degradation signaling compound covalently attached to the linker L, L is a linker that covalently attaches DSM to D, and D is a Bcl-xL inhibitor compound that is covalently linked to the linker. 1.
• Bcl-xL Inhibitors [118]
• the Bcl-xL inhibitor compound (D) of Formula (A) is represented by Formula (I) or Formula (II): , or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein the definitions of variables depicted in Formula (I) and (II) are described above (e.g., in the first or second embodiment).
• the Bcl-xL inhibitor compound (D) is represented by Formula (I), or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein R 1 is linear or branched C 1 -C 6 alkyl; R 2 is H; and the remaining variables are as described above for Formula (I).
• the Bcl-xL inhibitor compound (D) is represented by Formula (II), or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein A 4 and A 5 both represent a nitrogen atom, R 1 is linear or branched C 1-6 alkyl, R 2 is H, n is 1, ------ in Formula (II) represents a single bond, and the remaining variables are as described above for Formula (II).
• the Bcl-xL inhibitor compound (D) is represented by Formula (IA) or (IIA): , or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein: ⁇ Z 1 represents a bond or –O-, ⁇ R 3 represents a group selected from the group consisting of: hydrogen; a C 3 - C 6 cycloalkyl; a linear or branched C 1 -C 6 alkyl; -X 1 -NR a R b ; -X 1 -N + R a R b R c ; -X 1 -O-R c ; - X 1 -N 3 and , ⁇ R a and R b independently of one another represent a group selected from the group consisting of: hydrogen; a linear or branched C 1 -C 6 alkyl optionally substituted by one or two hydroxyl groups; and
• ⁇ A 1 is –NH-, -N(C 1 -C 3 alkyl), O, S or Se
• ⁇ A 2 is N, CH or C(R 5 )
• ⁇ R 8 represents a group selected from the group consisting of: hydrogen; a linear or branched C 1 -C 6 alkyl, -NR’ a R’ b ; -NR’ a -CO-OR’ c ; -NR’ a -CO-R’ c ; -N + R’ a R’ b R’ c ; -O-R’ c ; - NH-X’ 2 -N + R’ a R’ b R’ c ; -O-X’ 2 -NR’ a R’ b ; -X’ 2 -NR’ a R’ b ; -X’ 2 -NR’ a R’ b ; -NR’ c -X’ 2 -N 3 and: , ⁇ R 10 represents a group selected from the group consisting of hydrogen, fluorine, chlorine, bromine, -CF 3 and methyl, ⁇ R 11 represents a group selected from the group selected from the
• the Bcl-xL inhibitor compound (D) is represented by Formula (IA) or (IIA): , or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein: ⁇ Z 1 represents a bond or –O-, ⁇ R 3 represents a group selected from: hydrogen; C 3 -C 6 cycloalkyl; linear or branched C 1 -C 6 alkyl; -X 1 -NR a R b ; -X 1 -N + R a R b R c ; -X 1 -O-R c ; -X 1 -N 3 and , ⁇ R a and R b independently of one another represent a group selected from: hydrogen; linear or branched C 1 -C 6 alkyl optionally substituted by one or two hydroxyl groups; and C 1 -C 6 alkylene-SO 2 O-, ⁇
• R 7 represents a group selected from: linear or branched C 1 -C 6 alkyl group; (C 3 -C 6 )cycloalkylene-R 8 ; or: wherein Cy represents a C 3 -C 8 cycloalkyl.
• R 7 represents a group selected from: [125]
• the Bcl-xL inhbitor compound (D) is represented by Formula (IB), (IC-1), (IIB) or (IIC-1):
• R 3 represents a group selected from the group consisting of: hydrogen; a linear or branched C 1 -C 6 alkyl; -X 1 -NR a R b ; -X 1 -N + R a R b R c ; -X 1 -O-R c ; - X 1 -N 3 and , ⁇ for formula (IIB) or (IIC-1), Z 1 represents a bond, and R 3 represents hydrogen; or Z 1 represents –O-, and R 3 represents –X 1 -NR a R b , ⁇ for formula (IC-1), G is selected from the group consisting of –C(O)OH and - C(O)NR G1 R G2 ; ⁇ for formula (IIC-1), G is selected from the group consisting of
• R 3 represents a group selected from: hydrogen; linear or branched C 1 -C 6 alkyl; -X 1 -NR a R b ; -X 1 -N + R a R b R c ; -X 1 -O-R c ; -X 1 -N 3 and for formula (IIB) or (IIC), Z1 represents a bond, and R 3 represents hydrogen; or Z 1 represents –O-, and R 3 represents –X 1 -NR a R b , ⁇ R a and R b independently of one another represent a group selected from: hydrogen; linear or branched C 1 -C 6 alkyl optionally substituted by one or two hydroxyl groups; and C 1 -C 6 alkylene-SO 2 O-, ⁇ R c represents a hydrogen or a linear
• R 6 represents ⁇ X 2 -O-R 7 , and R 7 represents the following group: .
• R 6 represents a heteroarylene-R 7 group optionally substituted by a linear or branched C 1 -C 6 alkyl group, and R 7 represents a group selected from: .
• B 3 represents a C 3 -C 8 heterocycloalkyl group selected from a pyrrolidinyl group, a piperidinyl group, a piperazinyl group, a morpholinyl group, an azepanyl group, and a 4,4- difluoropiperidin-1-yl group.
• B 3 represents a pyrrolidinyl group or a piperazinyl group.
• B 3 represents a piperazinyl group.
• R 8 represents a group selected from the group consisting of: , wherein: is a bond to the linker.
• R 8 represents a group selected from: , wherein is a bond to the linker.
• R 8 represents a group selected from: , wherein is a bond to the linker.
• the Bcl-xL inhibitor compound (D) is represented by Formula (IB), (IIB), (IC), (IC-1), (IIC) or (IIC-1), or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein: ⁇ for formula (IB) or (IC-1), R 3 represents a group selected from the group consisting of: hydrogen; a linear or branched C 1 -C 6 alkyl; -X 1 -NR a R b ; -X 1 -N 3 and , ⁇ for formula (IIB) or (IIC-1), Z 1 represents a bond, and R 3 represents hydrogen, ⁇ for formula (IC-1), G is selected from the group consisting of –C(O)OH and - C(O)N(CH 3 ) 2 ; ⁇ for formula (IIC-1), G is selected from the group consisting of -C(O)NHS
• the Bcl-xL inhibitor compound (D) is represented by Formula (IB), (IIB), (IC), (IC-1), (IIC) or (IIC-1) , or an enantiomer, a diastereoisomer, and/or a pharmaceutically acceptable salt of any one of the foregoing, wherein: ⁇ for formula (IB) or (IC), R 3 represents a group selected from: hydrogen; linear or branched C 1 -C 6 alkyl; -X 1 -N 3 and for formula (IIB) or (IIC), Z 1 represents a bond, and R 3 represents hydrogen, ⁇ R 6 represents a linear or branched –C 1 -C 6 alkylene-R 8 group, –X 2 -O-R 7 or a heteroarylene-R 7 group optionally substituted by a linear or branched C 1 -C 6 alkyl group, ⁇ R 7 represents a group selected from: , ⁇ R 8 represents a group selected from:
• the Bcl-xL inhibitor compound (D) comprises a Bcl-xL inhibitor known in the art, for example, ABT-737 and ABT-263.
• D represents a Bcl-xL inhibitor attached to the linker L by a covalent bond, wherein the Bcl-xL inhibitor is selected from a compound in Table 1, or an enantiomer, a diastereomer and/or a pharmaceutically acceptable salt thereof.
• Table 1 Table 1
• D represents a moiety selected from any one of the formulae in Table 2, or an enantiomer, a diastereomer, and/or a pharmaceutically acceptable salt thereof, where represents a bond to the linker (L).
• Table 2 Exemplary Bcl-xL Moieties Showing Point of Attachment to the Linker (L)
• a bifuctional linker compound can be used to covalently attach a degradation signaling compound to a Bcl-xL inhibitor drug compound to form the PROTAC compounds of the present disclosure comprising a degradation signaling moiety (DSM) and a Bcl-xL inhibitor drug moiety (D).
• the bifunctional linker compound has at one end a reactive group that can react with the Bcl-xL inhibitor compound and at the other end another reactive group that can react with the degradation signaling compound.
• the bifunctional linker compound is reacted with the drug moiety (e.g., the Bcl-xL inhibitor) under appropriate conditions.
• a drug-linker compound is subsequently reacted with the degradation signaling compound, under conditions to form the PROTAC compound of the present disclosure.
• the linker compound can first react with the degradation signaling compound, to form a linker-DSM compound, which can then react with the drug to obtain the PROTAC compound of the present disclosure.
• the Bcl-xL inhibitor compound when the Bcl-xL inhibitor compound comprises a -NR’ c -X’ 2 -N 3 or group, it can react with an alkyne or an azide group of the linker compound to form a triazole group, which is considered to be part of the linker moiety L.
• the linker (L) comprises at least one group selected from the group consisting of: a linear or branched C 1 -C 20 alkylene optionally substituted by one to three groups selected from the group consisting of a C 1 -C 8 alkyl, a C 3 -C 8 cycloalkyl, trifluoromethyl, hydroxyl, a halogen, and a C 1 -C 6 alkoxy; a C 3 -C 1 0cycloalkylene; a C 3 -C 8 heterocycloalkylene; ⁇ C(O) ⁇ ; ⁇ O ⁇ ; ⁇ S ⁇ ; ⁇ N(R 16 ) ⁇ ; ⁇ N(R 16 )-C(O) ⁇ ; ⁇ C(O)-N(R 16 ) ⁇ ; ⁇ CH 2 ⁇ C(O) ⁇ N(R 16 ) ⁇ ; ⁇ N(R 16 ) ⁇ C(O) ⁇ CH 2 ⁇ ; a polyoxyethylene (PEG) group; an ethylene (PEG) group;
• the linker (L) comprises at least one group selected from: a linear or branched C 1 -C 20 alkylene optionally substituted by one or two groups selected from C 1 -C 8 alkyl, C 3 -C 8 cycloalkyl, trifluoromethyl, hydroxyl, halogen, and C 1 -C 6 alkoxy; a C 3 - C 1 0cyclo-alkylene; ⁇ C(O) ⁇ ; ⁇ O ⁇ ; ⁇ S ⁇ ; ⁇ N(R 16 ) ⁇ ; ⁇ N(R 16 )-C(O) ⁇ ; ⁇ C(O)-N(R 16 ) ⁇ ; ⁇ CH 2 ⁇ C(O) ⁇ N(R 16 ) ⁇ ; ⁇ N(R 16 ) ⁇ C(O) ⁇ CH 2 ⁇ ; a polyoxyethylene (PEG) group; an arylene group optionally substituted by one or two groups selected from C 1 -C 8 alkyl, trifluoromethyl, hydroxy
• the linker (L) comprises a 1,2,3-triazolene group formed by reacting an azide-containing precursor with an alkyne-containing precursor.
• the linker (L) is represented by formula (i): wherein: LK 1 is a bond, ⁇ NR 16 ⁇ or ⁇ C(O) ⁇ ; LK 2 is a bond, ⁇ C(O) ⁇ or ⁇ N(R 16 ) ⁇ C(O) ⁇ CH 2 ⁇ *; R 16 is H or methyl; R17 is a C 1 -C 20 alkylene, a C 3 -10cycloalkylene, a C 3-10 cycloalkylene-CH 2 -**, phenylene, –C 1 -C 20 alkylene-OCH 2 CH 2 -**, –C 1 -C 20 alkylene-OCH 2 -**,-CH 2 -(OCH 2 CH 2 )p- OCH 2 -** or -(CH 2 CH
• the linker (L) is represented by formula (i): wherein: LK 1 is a bond or ⁇ C(O) ⁇ ; LK 2 is a bond, ⁇ C(O) ⁇ or ⁇ N(R 16 ) ⁇ C(O) ⁇ CH 2 ⁇ *; R 16 is H or methyl; R 17 is C 1 -C 20 alkylene, C 3-10 cycloalkylene, phenylene, -CH 2 -(OCH 2 CH 2 ) p -OCH 2 -, wherein C 1 -C 15 alkylene or phenylene is optionally substituted with one or two R 17a ; p is an integer from 1 to 7; R 17a , for each occurrence, is independently a linear or branched C 1 -6alkyl or a halogen, or two R 17a together with the carbon atom from which they are attached form a C 3-6 cycloalkyl, and wherein: is a bond to the Bcl-xL
• the linker (L) is represented by formula (ii): wherein d is an integer from 1 to 7, and wherein: is a bond to the Bcl-xL inhibitor compound; and is a bond to DSM.
• the linker (L) is represented by formula (iii): wherein: LK 3 is ⁇ C(O) ⁇ or ⁇ N(R 16 ) ⁇ C(O) ⁇ CH 2 ⁇ *; R 16 is H or methyl; R 18 is C 1 -20alkylene or –CH 2 CH 2 -(OCH 2 CH 2 )p-**, wherein ** indicates the attachment point to LK 3 ; p is an integer from 1 to 7; and wherein: is a bond to the Bcl-xL inhibitor compound; and is a bond to DSM.
• the linker (L) is represented by formula (iv): , wherein: LK 4 is a bond or ⁇ C(O) ⁇ ; R 19 is C 1-6 alkylene; and wherein: is a bond to the Bcl- xL inhibitor compound; and is a bond to DSM.
• the linker (L) is represented by formula (v): wherein: LK 5 is a bond or ⁇ C(O) ⁇ ; R 20 is C 3-10 cycloalkylene, phenylene, ⁇ S ⁇ or ⁇ N(R 16 ) ⁇ ; R 16 is H or methyl; and wherein: is a bond to the Bcl-xL inhibitor compound; and is a bond to DSM.
• the linker (L) is represented by formula (ii): wherein: LK 6 is a bond, -C(O)-, ⁇ O ⁇ CH 2 ⁇ C(O) ⁇ *, or ⁇ N(R 16 ) ⁇ C(O) ⁇ CH 2 ⁇ *; R 16 is H or methyl; R 21 is C 1-20 alkylene or –CH 2 -(OCH 2 CH 2 ) p -**, wherein ** indicates the attachment point to LK 6 ; p is an integer from 1 to 7; and wherein: is a bond to the Bcl-xL inhibitor compound; and is a bond to DSM.
• LK 6 is a bond, ⁇ O ⁇ CH 2 ⁇ C(O) ⁇ *, or ⁇ N(R 16 ) ⁇ C(O) ⁇ CH 2 ⁇ *.
• the linker (L) is represented by formula (vii): wherein: LK 7 is a bond or ⁇ NR 16 ⁇ ; LK 8 is a bond, ⁇ R 22 ⁇ , ⁇ O ⁇ R 22 ⁇ or ⁇ C(O) ⁇ R 22 ⁇ ; Ring A is a C 3 -C 8 heterocyloalkylene; R 16 is H or methyl; R 22 is a C 1 -C 6 alkylene; is a bond to the Bcl-xL inhibitor compound; and is a bond to DSM.
• L is represented by a formula selected from formulae (L1)-(L109) in Table 3, where represents a bond to the Bcl-xL inhibitor compound (D), and represents a bond to the degradation signaling compound (DSM).
• Table 3 Exemplary Linkers Showing the Points of Attachment to the Bcl-xL Inhibitor Compound (D) and the Degradation Signaling Compound (DSM)
• D ⁇ L in Formula (A) is represented by a formula in Table 4, or an enantiomer, diastereoisomer and/or a pharmaceutically acceptable salt thereof, where represents a bond to the degradation signaling compound (DSM).
• Table 4 Exemplary D ⁇ L Moieties Showing the Point of Attachement to the Degradation Signaling Compound (DSM)
• Degradation signaling compounds and moieties (DSMs) of the present disclosure include compounds and moieties thereof that induce degradation of the targeted Bcl proteins (e.g., Bcl-xL). DSMs degrade Bcl by binding or recruiting at least one degradation protein, which is usually associated with the proteasome, the ubiquitin-proteasome pathways, or lysosomal proteolysis. DSMs of this disclosure include, but are not limited to, E3 ubiquitin ligase recognition agents.
• the E3 ubiquitin ligase or component of the E3 ubiquitin ligase complex targeted is MDM2, cIAPI, VHL protein, CBRN or SCF ⁇ -TRCP .
• the E3 ligase recognition agent is any compound that effectively binds to an E3 ubiquitin ligase or an E3 ubiquitin ligase complex.
• the E3 ligase recognition agent is an E3 ubiquitin ligase ligand, such as a VHL ligand, a thalidomide cereblon binder, or an inhibitor of apoptosis (IAP) E3 ligases.
• thalidomide cereblon binder refers to thalidomide or thalidomide derivatives (e.g., pomalidomide or a modified version of pomalidomide) that binds to cereblon.
• E3 ligase recognition agents are those described in WO 2021/007307, WO 2020/163823, US 2019/0127359, WO 2019/144117, WO 2018/200981, WO 2016/149668, WO 2016/105518, WO 2017/184995, WO 2017/007612, WO 2015/160845, Girardini, M.
• DSM represents a degradation signaling compound attached to the linker by a covalent bond, where a degradation signaling compound (DSM compound) is selected from a compound in Table 5, or an enantiomer, a diastereomer, and/or a pharmaceutically acceptable salt thereof.
• DSM compound a degradation signaling compound
• Table 5 Exemplary Degradation Signaling Compounds
• DSM in Formula (A) is represented by a formula in Table 6, or an enantiomer, a diastereomer and/or a pharmaceutically acceptable salt thereof, where represents a bond to the linker (L).
• DSM is DSM1a, or an enantiomer, a diastereomer, and/or a pharmaceutically acceptable salt thereof, where represents a bond to the linker (L): 4.
• Bifunctional Bcl-xL Degrader Compounds [166] In some embodiments, the Bcl-xL degrader compound is represented by Formula (A) described above (e.g., a compound described in any one of the first to twenty-eighth embodiments).
• the Bcl-xL degrader compound is a compound in Table 7 or an enantiomer, a diastereoisomer and/or a pharmaceutically acceptable salt thereof. [167] Table 7. Exemplary Bifunctional Bcl-xL Degrader Compounds
• compositions and Methods of Treatment are therapeutic uses of the disclosed compounds and compositions.
• An exemplary embodiment is an compound, composition, or pharmaceutical composition (e.g., any of the exemplary compounds, compositions, or pharmaceutical compositions disclosed herein) for use in treating a subject having or suspected of having a cancer (e.g., a Bcl-xL-mediated cancer).
• Another exemplary embodiment is a use of a compound, composition, or pharmaceutical composition (e.g., any of the exemplary compounds, compositions, or pharmaceutical compositions disclosed herein) in treating a subject having or suspected of having a cancer (e.g., a Bcl-xL-mediated cancer).
• Another exemplary embodiment is a use of a compound, composition, or pharmaceutical composition (e.g., any of the exemplary compounds, compositions, or pharmaceutical compositions disclosed herein) in a method of manufacturing a medicament for treating a subject having or suspected of having a cancer (e.g., a Bcl-xL-mediated cancer).
• the therapeutic compositions used in the practice of the foregoing methods may be formulated into pharmaceutical compositions comprising a pharmaceutically acceptable carrier suitable for the desired delivery method.
• An exemplary embodiment is a pharmaceutical composition comprising compound of the present disclosure and a pharmaceutically acceptable carrier, e.g., one suitable for a chosen means of administration, e.g., intravenous administration.
• the pharmaceutical composition may also comprise one or more additional inactive and/or therapeutic agents that are suitable for treating or preventing, for example, a cancer (e.g., a standard-of-care agent, etc.).
• the pharmaceutical composition may also comprise one or more carrier, excipient, and/or stabilizer components, and the like. Methods of formulating such pharmaceutical compositions and suitable formulations are known in the art (see, e.g., "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, PA).
• Suitable carriers include any material that, when combined with the therapeutic composition, retains the anti-tumor function of the therapeutic composition and is generally non-reactive with the patient's immune system.
• Pharmaceutically acceptable carriers include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible.
• examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, mesylate salt, and the like, as well as combinations thereof.
• isotonic agents are included, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition.
• compositions of the present disclosure can be administered by a variety of methods known in the art.
• the route and/or mode of administration may vary depending upon the desired results.
• the therapeutic formulation is solubilized and administered via any route capable of delivering the therapeutic composition to the cancer site.
• Potentially effective routes of administration include, but are not limited to, parenteral (e.g., intravenous, subcutaneous), intraperitoneal, intramuscular, intratumor, intradermal, intraorgan, orthotopic, and the like.
• the administration is intravenous, subcutaneous, intraperitoneal, or intramuscular.
• the pharmaceutically acceptable carrier should be suitable for the route of administration, e.g., intravenous or subcutaneous administration (e.g., by injection or infusion).
• the active compound(s) i.e., the compound and/or any additional therapeutic agent, may be coated in a material to protect the compound(s) from the action of acids and other natural conditions that may inactivate the compound(s).
• Administration can be either systemic or local.
• the therapeutic compositions disclosed herein may be sterile and stable under the conditions of manufacture and storage, and may be in a variety of forms.
• liquid, semi-solid, and solid dosage forms such as liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories.
• liquid solutions e.g., injectable and infusible solutions
• dispersions or suspensions tablets, pills, powders, liposomes, and suppositories.
• the form depends on the intended mode of administration and therapeutic application.
• the disclosed compounds can be incorporated into a pharmaceutical composition suitable for parenteral administration.
• the injectable solution may be composed of either a liquid or lyophilized dosage form in a flint or amber vial, ampule, or pre-filled syringe, or other known delivery or storage device.
• one or more of the compounds or pharmaceutical compositions is supplied as a dry sterilized lyophilized powder or water free concentrate in a hermetically sealed container and can be reconstituted (e.g., with water or saline) to the appropriate concentration for administration to a subject.
• a therapeutically effective amount or efficacious amount of a disclosed composition e.g., a disclosed compound
• the composition e.g., one comprising a compound disclosed herein, may be formulated into a pharmaceutically acceptable dosage form by conventional methods known in the art.
• Dosages and administration protocols for the treatment of cancers using the foregoing methods will vary with the method and the target cancer, and will generally depend on a number of other factors appreciated in the art.
• Dosage regimens for compositions disclosed herein e.g., those comprising compounds alone or in combination with at least one additional inactive and/or active therapeutic agent, may be adjusted to provide the optimum desired response (e.g., a therapeutic response). For example, a single bolus of one or both agents may be administered at one time, several divided doses may be administered over a predetermined period of time, or the dose of one or both agents may be proportionally increased or decreased as indicated by the exigencies of the therapeutic situation.
• treatment involves single bolus or repeated administration of the compound preparation via an acceptable route of administration.
• the compound is administered to the patient daily, weekly, monthly, or any time period in between.
• specific dosage regimens may be adjusted over time according to the individual’s need, and the professional judgment of the treating clinician.
• Parenteral compositions may be formulated in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit contains a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.
• Dosage values for compositions comprising compounds disclosed herein and/or any additional therapeutic agent(s), may be selected based on the unique characteristics of the active compound(s), and the particular therapeutic effect to be achieved.
• a physician or veterinarian can start doses of the compound employed in the pharmaceutical composition at levels lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.
• effective doses of the compositions of the present disclosure, for the treatment of a cancer may vary depending upon many different factors, including means of administration, target site, physiological state of the patient, whether the patient is human or an animal, other medications administered, and whether treatment is prophylactic or therapeutic.
• the selected dosage level may also depend upon a variety of pharmacokinetic factors including the activity of the particular compositions of the present disclosure employed, or the ester, salt, or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compositions employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors. Treatment dosages may be titrated to optimize safety and efficacy. [176] Toxicity and therapeutic efficacy of compounds provided herein can be determined by standard pharmaceutical procedures in cell culture or in animal models.
• LD50, ED50, EC50, and IC50 may be determined, and the dose ratio between toxic and therapeutic effects (LD50/ED50) may be calculated as the therapeutic index.
• the data obtained from in vitro and in vivo assays can be used in estimating or formulating a range of dosage for use in humans.
• the compositions and methods disclosed herein may initially be evaluated in xenogeneic cancer models (e.g., an NCI-H929 multiple myeloma mouse model).
• a compound disclosed herein or a composition comprising a compound is administered on a single occasion. In other embodiments, a compound or composition comprising the compound is administered on multiple occasions.
• Intervals between single dosages can be, e.g., daily, weekly, monthly, or yearly. Intervals can also be irregular, based on measuring blood levels of the administered agent (e.g., the compound) in the patient in order to maintain a relatively consistent plasma concentration of the agent.
• the dosage and frequency of administration of a compound or composition comprising the compound may also vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic applications, a relatively low dosage may be administered at relatively infrequent intervals over a long period of time. Some patients continue to receive treatment for the rest of their lives. In therapeutic applications, a relatively higher dosage at relatively shorter intervals is sometimes required until progression of the disease is reduced or terminated, and preferably until the patient shows partial or complete amelioration of one or more symptoms of disease.
• compounds of the present disclosure may be administered in an amount effective to sensitize tumor cells to one or more additional therapeutic agents and/or radiation therapy.
• compounds of the present disclosure may be administered as monotherapy, while in other embodiments the compounds may be administered adjunctive to another therapeutic agent or radiation therapy.
• methods of the present disclosure involve the further administration (in addition of at least one PROTAC compound disclosed herein) to a subject in need thereof at least one additional therapeutic agent—such as, for example, a Bcl-2 inhibitor, a taxane, a MEK inhibitor, an ERK inhibitor, or a RAF inhibitor.
• kits for use in the therapeutic and/or diagnostic applications described herein are also provided. Such kits may comprise a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in a method disclosed herein.
• a label may be present on or with the container(s) to indicate that a compound or composition within the kit is used for a specific therapy or non-therapeutic application, such as a prognostic, prophylactic, diagnostic, or laboratory application.
• a label may also indicate directions for either in vivo or in vitro use, such as those described herein. Directions and or other information may also be included on an insert(s) or label(s), which is included with or on the kit.
• the label may be on or associated with the container.
• a label may be on a container when letters, numbers, or other characters forming the label are molded or etched into the container itself.
• a label may be associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert.
• the label may indicate that the compound or composition within the kit is used for diagnosing or treating a condition, such as a cancer a described herein.
• a kit comprises a compound or composition comprising the compound.
• the kit further comprises one or more additional components, including but not limited to: instructions for use; other reagents, e.g., a therapeutic agent (e.g., a standard-of-care agent); devices, containers, or other materials for preparing the compound for administration; pharmaceutically acceptable carriers; and devices, containers, or other materials for administering the compound to a subject.
• kits for use can include guidance for therapeutic applications including suggested dosages and/or modes of administration, e.g., in a patient having or suspected of having a cancer.
• the kit comprises a compound and instructions for use of the compound in treating, preventing, and/or diagnosing a cancer.
• elevated Bcl-xL expression correlates with resistance to radiation therapy and chemotherapy.
• Compounds of the present disclosure that may not be sufficiently effective as monotherapy to treat cancer can be administered in combination with other therapeutic agents (including non-targeted and targeted therapeutic agents) or radiation therapy (including radioligand therapy) to provide therapeutic benefit.
• the linked-drug conjugates described herein may sensitize tumor cells to the treatment with other therapeutic agents (including standard of care chemotherapeutic agents to which the tumor cells may have developed resistance) and/or radiation therapy.
• compounds described herein are administered to a subject having cancer in an amount effective to sensitize the tumor cells.
• the term “sensitize” means that the treatment with the compound increases the potency or efficacy of the treatment with other therapeutic agents and/or radiation therapy against tumor cells.
• Compound and compositions of the present disclosure may be administered alone or in combination with at least one additional inactive and/or active agent, e.g., at least one additional therapeutic agent, and may be administered in any pharmaceutically acceptable formulation, dosage, and dosing regimen.
• Treatment efficacy may be evaluated for toxicity as well as indicators of efficacy and adjusted accordingly.
• Efficacy measures include, but are not limited to, a cytostatic and/or cytotoxic effect observed in vitro or in vivo, reduced tumor volume, tumor growth inhibition, and/or prolonged survival.
• the present disclosure features a method of killing, inhibiting or modulating the growth of a cancer cell or tissue by disrupting the expression and/or activity of Bcl-xL and/or one or more upstream modulators or downstream targets thereof.
• the method may be used with any subject where disruption of Bcl-xL expression and/or activity provides a therapeutic benefit.
• Subjects that may benefit from disrupting Bcl-xL expression and/or activity include, but are not limited to, those having or at risk of having a cancer such as a tumor or a hematological cancer.
• the cancer is a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular cancer, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or spleen cancer.
• the cancer is a lymphoma or gastric cancer.
• Exemplary methods include the steps of contacting a cell with a compound or composition described herein, in an effective amount, i.e., an amount sufficient to kill the cell.
• the method can be used on cells in culture, e.g., in vitro, in vivo, ex vivo, or in situ.
• cells e.g., cells collected by biopsy of a tumor or metastatic lesion; cells from an established cancer cell line; or recombinant cells
• the contacting step can be affected by adding the compound or composition to the culture medium.
• the compound or composition can be administered to a subject by any suitable administration route (e.g., intravenous, subcutaneous, or direct contact with a tumor tissue) to have an effect in vivo.
• a suitable administration route e.g., intravenous, subcutaneous, or direct contact with a tumor tissue
• the in vivo effect of a therapeutic composition disclosed herein can be evaluated in a suitable animal model.
• xenogeneic cancer models can be used, wherein cancer explants or passaged xenograft tissues are introduced into immune compromised animals, such as nude or SCID mice (Klein et al. (1997) Nature Med. 3:402-8). Efficacy may be predicted using assays that measure inhibition of tumor formation, tumor regression or metastasis, and the like.
• xenografts from tumor bearing mice treated with the therapeutic composition can be examined for the presence of apoptotic foci and compared to untreated control xenograft-bearing mice. The extent to which apoptotic foci are found in the tumors of the treated mice provides an indication of the therapeutic efficacy of the composition.
• a disorder e.g., a cancer.
• the compositions described herein can be administered to a non-human mammal or human subject for therapeutic purposes.
• the therapeutic methods include administering to a subject having or suspected of having a cancer a therapeutically effective amount of a composition comprising an Bcl-xL inhibitor.
• An exemplary embodiment is a method of treating a subject having or suspected of having a cancer, comprising administering to the subject a therapeutically effective amount of a composition disclosed herein.
• the cancer is a solid tumor or a hematological cancer.
• the cancer is a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia colorectal cancer esophageal cancer hepatocellular cancer lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or spleen cancer.
• the cancer is a lymphoma or gastric cancer.
• An exemplary embodiment is a method of reducing or inhibiting the growth of a tumor in a subject, comprising administering to the subject a therapeutically effective amount of a PROTAC compound, composition, or pharmaceutical composition (e.g., any of the exemplary compounds, compositions, or pharmaceutical compositions disclosed herein).
• administration of the compound, composition, or pharmaceutical composition reduces or inhibits the growth of the tumor by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%, as compared to growth in the absence of treatment.
• the present disclosure further provides methods of reducing or slowing the expansion of a cancer cell population comprising administering a therapeutically effective amount of a PROTAC compound or composition comprising a PROTAC compound.
• the compounds or compositions of the present disclosure may be administered to a non-human mammal for veterinary purposes or as an animal model of human disease.
• a combination described herein comprises a PD-1 inhibitor.
• the PD-1 inhibitor is chosen from PDR001 (Novartis), Nivolumab (Bristol- Myers Squibb), Pembrolizumab (Merck & Co), Pidilizumab (CureTech), MEDI0680 (Medimmune), REGN2810 (Regeneron), TSR-042 (Tesaro), PF-06801591 (Pfizer), BGB- A317 (Beigene), BGB-108 (Beigene), INCSHR1210 (Incyte), or AMP-224 (Amplimmune).
• the PD-1 inhibitor is PDR001.
• PDR001 is also known as Spartalizumab.
• a combination described herein comprises a LAG-3 inhibitor.
• the LAG-3 inhibitor is chosen from LAG525 (Novartis), BMS-986016 (Bristol-Myers Squibb), or TSR-033 (Tesaro).
• a combination described herein comprises a TIM-3 inhibitor.
• the TIM-3 inhibitor is MBG453 (Novartis), TSR-022 (Tesaro), LY- 3321367 (Eli Lily), Sym23 (Symphogen), BGB-A425 (Beigene), INCAGN-2390 (Agenus), BMS-986258 (BMS), RO-7121661 (Roche), or LY-3415244 (Eli Lilly).
• a combination descdribed herein comprises a PDL1 inhibitor.
• the PDL1 inhibitor is chosen from FAZ053 (Novartis), atezolizumab (Genentech), durvalumab (Astra Zeneca), or avelumab (Pfizer).
• a combination described herein comprises a GITR agonist.
• the GITR agonist is chosen from GWN323 (NVS), BMS-986156, MK- 4166 or MK-1248 (Merck), TRX518 (Leap Therapeutics), INCAGN1876 (Incyte/Agenus), AMG 228 (Amgen) or INBRX-110 (Inhibrx).
• a combination described herein comprises an IAP inhibitor.
• the IAP inhibitor comprises LCL161 or a compound disclosed in International Application Publication No. WO 2008/016893.
• the combination comprises an mTOR inhibitor, e.g., RAD001 (also known as everolimus).
• the combination comprises a HDAC inhibitor, e.g., LBH589. LBH589 is also known as panobinostat.
• the combination comprises an IL-17 inhibitor, e.g., CJM112.
• a combination described herein comprises an estrogen receptor (ER) antagonist.
• the estrogen receptor antagonist is used in combination with a PD-1 inhibitor, a CDK4/6 inhibitor, or both.
• the combination is used to treat an ER positive (ER+) cancer or a breast cancer (e.g., an ER+ breast cancer).
• the estrogen receptor antagonist is a selective estrogen receptor degrader (SERD).
• SESDs are estrogen receptor antagonists which bind to the receptor and result in e.g., degradation or down-regulation of the receptor (Boer K. et al., (2017) Therapeutic Advances in Medical Oncology 9(7): 465-479).
• ER is a hormone-activated transcription factor important for e.g., the growth, development and physiology of the human reproductive system. ER is activated by, e.g., the hormone estrogen (17beta estradiol).
• the SERD is chosen from LSZ102, fulvestrant, brilanestrant, or elacestrant.
• the SERD comprises a compound disclosed in International Application Publication No. WO 2014/130310, which is hereby incorporated by reference in its entirety.
• the SERD comprises LSZ102.
• the LSZ102 has the chemical name: (E)-3-(4-((2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-hydroxybenzo[b]thiophen-3- yl)oxy)phenyl)acrylic acid.
• the SERD comprises fulvestrant (CAS Registry Number: 129453-61-8), or a compound disclosed in International Application Publication No. WO 2001/051056, which is hereby incorporated by reference in its entirety.
• the SERD comprises elacestrant (CAS Registry Number: 722533-56-4), or a compound disclosed in U.S. Patent No.7,612,114, which is incorporated by reference in its entirety.
• Elacestrant is also known as RAD1901, ER-306323 or (6R)-6- ⁇ 2-[Ethyl( ⁇ 4-[2- (ethylamino)ethyl]phenyl ⁇ methyl)amino]-4-methoxyphenyl ⁇ -5,6,7,8-tetrahydronaphthalen-2- ol.
• Elacestrant is an orally bioavailable, non-steroidal combined selective estrogens receptor modulator (SERM) and a SERD.
• SERM selective estrogens receptor modulator
• Elacestrant is also disclosed, e.g., in Garner F et al., (2015) Anticancer Drugs 26(9):948-56.
• the SERD is brilanestrant (CAS Registry Number: 1365888-06-7), or a compound disclosed in International Application Publication No. WO 2015/136017, which is incorporated by reference in its entirety.
• the SERD is chosen from RU 58668, GW7604, AZD9496, apeledoxifene, pipendoxifene, arzoxifene, OP-1074, or acolbifene, e.g., as disclosed in McDonell et al. (2015) Journal of Medicinal Chemistry 58(12) 4883-4887.
• a combination described herein comprises an inhibitor of Cyclin-Dependent Kinases 4 or 6 (CDK4/6).
• CDK4/6 Cyclin-Dependent Kinases 4 or 6
• the CDK4/6 inhibitor is used in combination with a PD-1 inhibitor, an estrogen receptor (ER) antagonist, or both.
• the combination is used to treat an ER positive (ER+) cancer or a breast cancer (e.g., an ER+ breast cancer).
• the CDK4/6 inhibitor is chosen from ribociclib, abemaciclib (Eli Lilly), or palbociclib.
• the CDK4/6 inhibitor comprises ribociclib (CAS Registry Number: 1211441-98-3), or a compound disclosed in U.S. Patent Nos. 8,415,355 and 8,685,980, which are incorporated by reference in their entirety.
• the CDK4/6 inhibitor comprises a compound disclosed in International Application Publication No. WO 2010/020675 and U.S. Patent Nos. 8,415,355 and 8,685,980, which are incorporated by reference in their entirety.
• the CDK4/6 inhibitor comprises ribociclib (CAS Registry Number: 1211441-98-3). Ribociclib is also known as LEE011, KISQALI®, or 7-cyclopentyl- N,N-dimethyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6- carboxamide.
• the CDK4/6 inhibitor comprises abemaciclib (CAS Registry Number: 1231929-97-7).
• Abemaciclib is also known as LY835219 or N-[5-[(4-Ethyl-1- piperazinyl)methyl]-2-pyridinyl]-5-fluoro-4-[4-fluoro-2-methyl-1-(1-methylethyl)-1H- benzimidazol-6-yl]-2-pyrimidinamine.
• Abemaciclib is a CDK inhibitor selective for CDK4 and CDK6 and is disclosed, e.g., in Torres-Guzman R et al. (2017) Oncotarget 10.18632/oncotarget.17778. [215]
• the CDK4/6 inhibitor comprises palbociclib (CAS Registry Number: 571190-30-2).
• Palbociclib is also known as PD-0332991, IBRANCE® or 6-Acetyl-8- cyclopentyl-5-methyl-2- ⁇ [5-(1-piperazinyl)-2-pyridinyl]amino ⁇ pyrido[2,3-d]pyrimidin-7(8H)- one.
• Palbociclib inhibits CDK4 with an IC50 of 11nM, and inhibits CDK6 with an IC50 of 16nM, and is disclosed, e.g., in Finn et al. (2009) Breast Cancer Research 11(5):R77.
• a combination described herein comprises an inhibitor of chemokine (C-X-C motif) receptor 2 (CXCR2).
• the CXCR2 inhibitor is chosen from 6-chloro-3-((3,4-dioxo-2-(pentan-3-ylamino)cyclobut-1-en-1-yl)amino)-2- hydroxy-N-methoxy-N-methylbenzenesulfonamide, danirixin, reparixin, or navarixin.
• the CSF-1/1R binding agent is chosen from an inhibitor of macrophage colony-stimulating factor (M-CSF), e.g., a monoclonal antibody or Fab to M-CSF (e.g., MCS110), a CSF-1R tyrosine kinase inhibitor (e.g., 4-((2-(((1R,2R)-2- hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide or BLZ945), a receptor tyrosine kinase inhibitor (RTK) (e.g., pexidartinib), or an antibody targeting CSF-1R (e.g., emactuzumab or FPA008).
• M-CSF macrophage colony-stimulating factor
• MCS110 monoclonal antibody or Fab to M-CSF
• CSF-1R tyrosine kinase inhibitor e
• the CSF-1/1R inhibitor is BLZ945.
• the CSF-1/1R binding agent is MCS110.
• the CSF- 1/1R binding agent is pexidartinib.
• a combination described herein comprises a c-MET inhibitor.
• c-MET a receptor tyrosine kinase overexpressed or mutated in many tumor cell types, plays key roles in tumor cell proliferation, survival, invasion, metastasis, and tumor angiogenesis. Inhibition of c-MET may induce cell death in tumor cells overexpressing c-MET protein or expressing constitutively activated c-MET protein.
• the c-MET inhibitor is chosen from capmatinib (INC280), JNJ-3887605, AMG 337, LY2801653, MSC2156119J, crizotinib, tivantinib, or golvatinib.
• a combination described herein comprises a transforming growth factor beta (also known as TGF- ⁇ TGF ⁇ , TGFb, or TGF-beta, used interchangeably herein) inhibitor.
• the TGF- ⁇ inhibitor is chosen from fresolimumab or XOMA 089.
• a combination described herein comprises an adenosine A2a receptor (A2aR) antagonist (e.g., an inhibitor of A2aR pathway, e.g., an adenosine inhibitor, e.g., an inhibitor of A2aR or CD-73).
• A2aR antagonist is used in combination with a PD-1 inhibitor, and one or more (e.g., two, three, four, five, or all) of a CXCR2 inhibitor, a CSF-1/1R binding agent, LAG-3 inhibitor, a GITR agonist, a c-MET inhibitor, or an IDO inhibitor.
• the combination is used to treat a pancreatic cancer, a colorectal cancer, a gastric cancer, or a melanoma (e.g., a refractory melanoma).
• the A2aR antagonist is chosen from PBF509 (NIR178) (Palobiofarma/Novartis), CPI444/V81444 (Corvus/Genentech), AZD4635/HTL-1071 (AstraZeneca/Heptares) Vipadenant (Redox/Juno) GBV-2034 (Globavir) AB928 (Arcus Biosciences), Theophylline, Istradefylline (Kyowa Hakko Kogyo), Tozadenant/SYN-115 (Acorda), KW-6356 (Kyowa Hakko Kogyo), ST-4206 (Leadiant Biosciences), or Preladenant/SCH 420814 (Merck/Scher
• a combination described herein comprises an inhibitor of indoleamine 2,3-dioxygenase (IDO) and/or tryptophan 2,3-dioxygenase (TDO).
• IDO indoleamine 2,3-dioxygenase
• TDO tryptophan 2,3-dioxygenase
• the IDO inhibitor is used in combination with a PD-1 inhibitor, and one or more (e.g., two, three, four, or all) of a TGF- ⁇ inhibitor, an A2aR antagonist, a CSF-1/1R binding agent, a c-MET inhibitor, or a GITR agonist.
• the combination is used to treat a pancreatic cancer, a colorectal cancer, a gastric cancer, or a melanoma (e.g., a refractory melanoma).
• the IDO inhibitor is chosen from (4E)-4-[(3- chloro-4-fluoroanilino)-nitrosomethylidene]-1,2,5-oxadiazol-3-amine (also known as epacadostat or INCB24360), indoximod (NLG8189), (1-methyl-D-tryptophan), ⁇ -cyclohexyl- 5H-Imidazo[5,1-a]isoindole-5-ethanol (also known as NLG919), indoximod, BMS-986205 (formerly F001287).
• a combination described herein comprises a Galectin, e.g., Galectin-1 or Galectin-3, inhibitor.
• the combination comprises a Galectin-1 inhibitor and a Galectin-3 inhibitor.
• the combination comprises a bispecific inhibitor (e.g., a bispecific antibody molecule) targeting both Galectin- 1 and Galectin-3.
• the Galectin inhibitor is used in combination with one or more therapeutic agents described herein.
• the Galectin inhibitor is chosen from an anti-Galectin antibody molecule, GR-MD-02 (Galectin Therapeutics), Galectin-3C (Mandal Med), Anginex, or OTX-008 (OncoEthix, Merck).
• a combination described herein comprises an inhibitor of the MAP kinase pathway including ERK inhibitors, MEK inhibitors and RAF inhibitors.
• a combination described herein comprises a MEK inhibitor.
• the MEK inhibitor is chosen from Trametinib, selumetinib, AS703026, BIX 02189, BIX 02188, CI-1040, PD0325901, PD98059, U0126, XL-518, G-38963, or G02443714. [225] In some embodiments, the MEK inhibitor is trametinib.
• Trametinib is also known as JTP- 74057, TMT212, N-(3- ⁇ 3-cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7- trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidin-1(2H)-yl ⁇ phenyl)acetamide, or Mekinist (CAS Number 871700-17-3).
• the MEK inhibitor comprises selumetinib which has the chemical name: (5-[(4-bromo-2-chlorophenyl)amino]-4-fluoro-N-(2-hydroxyethoxy)-1-methyl-1H- benzimid azole-6-carboxamide.
• Selumetinib is also known as AZD6244 or ARRY 142886, e.g., as described in PCT Publication No. WO2003077914.
• the MEK inhibitor comprises AS703026, BIX 02189 or BIX 02188.
• the MEK inhibitor comprises 2-[(2-Chloro-4-iodophenyl)amino]- N-(cyclopropylmethoxy)-3,4-difluoro-benzamide (also known as CI-1040 or PD184352), e.g., as described in PCT Publication No. WO2000035436.
• the MEK inhibitor comprises N-[(2R)-2,3-Dihydroxypropoxy]- 3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]- benzamide (also known as PD0325901), e.g., as described in PCT Publication No.
• the MEK inhibitor comprises 2’-amino-3’-methoxyflavone (also known as PD98059) which is available from Biaffin GmbH & Co., KG, Germany.
• the MEK inhibitor comprises 2,3-bis[amino[(2- aminophenyl)thio]methylene]-butanedinitrile (also known as U0126), e.g., as described in US Patent No.2,779,780.
• the MEK inhibitor comprises XL-518 (also known as GDC-0973) which has a CAS No.1029872-29-4 and is available from ACC Corp.
• the MEK inhibitor comprises G-38963. In some embodiments, the MEK inhibitor comprises G02443714 (also known as AS703206). [234] Additional examples of MEK inhibitors are disclosed in WO 2013/019906, WO 03/077914, WO 2005/121142, WO 2007/04415, WO 2008/024725 and WO 2009/085983, the contents of which are incorporated herein by reference. Further examples of MEK inhibitors include, but are not limited to, 2,3-Bis[amino[(2-aminophenyl)thio]methylene]-butanedinitrile (also known as U0126 and described in US Patent No.
• a combination described herein comprises a RAF inhibitor.
• RAF inhibitors include, but are not limited to, Vemurafenib (or Zelboraf®, PLX-4032, CAS 918504-65-1), GDC-0879, PLX-4720 (available from Symansis), Dabrafenib (or GSK2118436), LGX 818, CEP-32496, UI-152, RAF 265, Regorafenib (BAY 73-4506), CCT239065, or Sorafenib (or Sorafenib Tosylate, or Nexavar®).
• the RAF inhibitor is Dabrafenib.
• the RAF inhibitor is LXH254.
• a combination described herein comprises an ERK inhibitor.
• ERK inhibitors include, but are not limited to, LTT462, ulixertinib (BVD-523), LY3214996, GDC-0994, KO-947 and MK-8353.
• the ERK inhibitor is LTT462.
• LTT462 is 4-(3-amino-6- ((1S,3S,4S)-3-fluoro-4-hydroxy ⁇ cyclohexyl)pyrazin-2-yl)-N-((S)-1-(3-bromo-5-fluorophenyl)- 2-(methylamino)ethyl)-2-fluorobenzamide and is the compound of the following structure: [242]
• the preparation of LTT462 is described in PCT patent application publication WO2015/066188.
• LTT462 is an inhibitor of extracellular signal-regulated kinases 1 and 2 (ERK 1/2).
• a combination described herein comprises a taxane, a MEK inhibitor, an ERK inhibitor, or a RAF inhibitor.
• a combination described herein comprises at least two inhibitors selected, independently, from a MEK inhibitor, an ERK inhibitor, and a RAF inhibitor.
• a combination described herein comprises an anti-mitotic drug.
• a combination described herein comprises a taxane.
• Taxanes include, but are not limited to, docetaxel, paclitaxel, or cabazitaxel. In some embodiments, the taxane is docetaxel.
• a combination described herein comprises a topoisomerase inhibitor.
• Topoisomerase inhibitors include, but are not limited to, topotecan, irinotecan, camptothecin, diflomotecan, lamellarin D, ellipticines, etoposide (VP-16), teniposide, doxorubicin, daunorubicin, mitoxantrone, amsacrine, aurintricarboxylic acid, and HU-331.
• a combination described herein includes an interleukin-1 beta (IL- 1 ⁇ ) inhibitor.
• the IL-1 ⁇ inhibitor is chosen from canakinumab, gevokizumab, Anakinra, or R i lonacept.
• a combination described herein comprises an IL-15/IL-15Ra complex.
• the IL-15/IL-15Ra complex is chosen from NIZ985 (Novartis), ATL-803 (Altor) or CYP0150 (Cytune).
• a combination described herein comprises a mouse double minute 2 homolog (MDM2) inhibitor.
• the human homolog of MDM2 is also known as HDM2.
• an MDM2 inhibitor described herein is also known as a HDM2 inhibitor.
• the MDM2 inhibitor is chosen from HDM201 or CGM097.
• the MDM2 inhibitor comprises (S)-1-(4-chlorophenyl)-7-isopropoxy- 6-methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3-oxopiperazin-1- yl)cyclohexyl)methyl)amino)phenyl)-12-dihydroisoquinolin-3(4H)-one (also known as CGM097) or a compound disclosed in PCT Publication No. WO 2011/076786 to treat a disorder, e.g., a disorder described herein).
• a therapeutic agent disclosed herein is used in combination with CGM097.
• a combination described herein comprises a hypomethylating agent (HMA).
• HMA hypomethylating agent
• the HMA is chosen from decitabine or azacitidine.
• a combination described herein comprises an inhibitor acting on any pro-survival proteins of the Bcl2 family.
• a combination described herein comprises a Bcl-2 inhibitor.
• the Bcl-2 inhibitor is venetoclax (venetoclax).
• the Bcl-2 inhibitor is selected from the compounds described in WO 2013/110890 and WO 2015/011400.
• the Bcl-2 inhibitor comprises navitoclax (ABT-263), ABT-737, BP1002, SPC2996, APG-1252, obatoclax mesylate (GX15- 070MS), PNT2258, Zn-d5, BGB-11417, or oblimersen (G3139).
• the Bcl-2 inhibitor is N-(4-hydroxyphenyl)-3-[6-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H- isoquinoline-2-carbonyl]-1,3-benzodioxol-5-yl]-N-phenyl-5,6,7,8-tetrahydroindolizine-1- carboxamide, compound A1: (compound A1).
• the Bcl-2 inhibitor is (S)-5-(5-chloro-2-(3-(morpholinomethyl)- 1,2,3,4-tetrahydroisoquinoline-2-carbonyl)phenyl)-N-(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl)-N- (4-hydroxyphenyl)-1,2-dimethyl-1H-pyrrole-3-carboxamide), compound A2: (compound A2).
• the DSM-drug conjugates or combinations disclosed herein are suitable for the treatment of cancer in vivo.
• the combination can be used to inhibit the growth of cancerous tumors.
• the combination can also be used in combination with one or more of: a standard of care treatment (e.g., for cancers or infectious disorders), a vaccine (e.g., a therapeutic cancer vaccine), a cell therapy, a hormone therapy (e.g., with anti- estrogens or anti-androgens), a radiation therapy, surgery, or any other therapeutic agent or modality, to treat a disorder herein.
• a standard of care treatment e.g., for cancers or infectious disorders
• a vaccine e.g., a therapeutic cancer vaccine
• a cell therapy e.g., a hormone therapy (e.g., with anti- estrogens or anti-androgens)
• a radiation therapy e.g., surgery, or any other therapeutic agent or modality
• the combination can be administered together with an antigen of interest.
• a combination disclosed herein can be administered in either order or simultaneously.
• Microwave Heating Microwave heating was performed in CEM Discover® instrument, or with an Anton Paar Monowave Microwave Reactor.
• NMR 1 H-NMR measurements were performed on 400 MHz Bruker® Avance or 500 MHz Avance Neo spectrometer, using DMSO-d6 or CDCl3 as solvent. 1 H NMR data is in the form of delta values, given in part per million (ppm), using the residual peak of the solvent (2.50 ppm for DMSO-d6 and 7.26 ppm for CDCl3) as internal standard.
• Splitting patterns are designated as: s (singlet), d (doublet), t (triplet), q (quartet), quint (quintet), m (multiplet), br s (broad singlet), dd (doublet of doublets), td (triplet of doublets), dt (doublet of triplets), ddd (doublet of doublet of doublets).
• IR IR measurements were performed on a Bruker® Tensor 27 equipped with ATR Golden Gate® device (SPECAC).
• Mass Spectrometry High-Resolution MS measurements (HRMS) were performed on a LTQ OrbiTrap® Velos Pro mass spectrometer (ThermoFisher Scientific).
• Jet Stream parameters drying gas (N 2 ) flow and temperature: 10.0 l/min and 300 °C, respectively; nebulizer gas (N 2 ) pressure: 40 psi; capillary voltage: 2500 V; sheath gas flow and temperature: 300 °C and 10.0 l/min; QTOFMS parameters: fragmentor voltage: 100 V; skimmer potential: 65 V; OCT 1 RF Vpp:750 V.
• UPLC®-MS [267] UPLC®-MS data were acquired using an instrument with the following parameters (Table 8): Table 8. UPLC®-MS Parameters Preparative HPLC (Prep-HPLC): [268] Preparative HPLC (“Prep-HPLC”) data were acquired using an instrument with the parameters of Table 9, or using an instrument with the parameteres of Table 10: Table 9. Prep-HPLC Parameters (Interchim Method) Table 10.
• Bcl-xL payloads were synthesized using exemplary methods described in this example.
• Mitsunobu General Procedure To the mixture of 1.0-1.5 eq. of aliphatic alcohol, 1 eq. of carbamate/phenol, and 1-2 eq. triphenylphosphine in THF or toluene (5 mL/mmol) were added 1-3 eq. of ditertbutyl azodicarboxylate / diisopropyl azodicarboxylate in one portion.
• Step B [(hex-4-yn-1-yloxy)methyl]benzene [280]
• the product from Step A (19.5 g, 112 mmol, 1 eq) and tetrahydrofuran (200 mL) and the solution was cooled to -78 °C.
• n-Butyllithium (2M solution in hexanes, 66.9 mL, 135 mmol, 1.2 eq) was added dropwise over 30 min and the reaction was stirred for 1 h then iodomethane (10.5 mL, 168 mmol, 1.5 eq) was added dropwise and the mixture was allowed to warm to 0 °C over 1 h.
• reaction was quenched by the addition of saturated aqueous ammonium chloride (40 mL), diluted with water (40 mL), extracted with ethyl acetate (3 x 100 mL), and the combined organic extracts were successively washed with 2M aqueous sodium thiosulfate (200 mL) and brine (200 mL), dried (magnesium sulfate) and concentrated in vacuo.
• Step C 4-[3-(benzyloxy)propyl]-3,6-dichloro-5-methylpyridazine [281]
• a solution of 3,6-dichloro-1,2,4,5-tetrazine (5 g, 33.1 mmol, 1 eq) and the product from Step B (7.48 g, 39.8 mmol, 1.2 eq) in tetrahydrofuran (30 mL) was heated at 160°C for 19 h in a sealed flask. The reaction was allowed to cool to ambient temperature then concentrated in vacuo.
• Step D 3-(3,6-dichloro-5-methyl-pyridazin-4-yl)propan-1-ol [282]
• dichloromethane 100 mL
• boron trichloride solution 1 M in dichloromethane; 58.8 mL, 58.8 mmol, 2.5 eq
• the reaction was quenched by the addition of methanol and concentrated in vacuo.
• Step B methyl 3-bromo-6-(tert-butoxycarbonylamino)pyridine-2-carboxylate [284] To the product from Step A (42.7 g, 74.34 mmol) in DCM (370 mL) was added TFA (17.1 mL, 3 eq) at 0°C and the reaction mixture was stirred for 18 h.
• Step C methyl 3-bromo-6-[tert-butoxycarbonyl-[3-(3,6-dichloro-5-methyl-pyridazin -4- yl)propyl]amino]pyridine-2-carboxylate [285] After mixing the product of Step B (748 mg, 2.2 mmol), the product of Preparation 1 (500 mg, 1 eq), and PPh 3 (593 mg, 1 eq) in toluene (10 mL), DTAD (520 mg, 1 eq) was added, and stirred at 50°C for 30 min.
• DTAD 520 mg, 1 eq
• Step D methyl 3-bromo-6-[3-(3,6-dichloro-5-methyl-pyridazin-4-yl)propylamino] pyridine-2-carboxylate
• the product from Step C (17.5 g, 32.7 mmol) in 1,1,1,3,3,3-hexafluoroisopropanol (330 mL) was stirred at 110°C for 18 h. Purification by column chromatography (silica gel, heptane and EtOAc as eluents) afforded the desired product (9.9 g, 70%).
• Step B 3-bromo-5,7-dimethyl-1-adamantyl-methanol [288] To the product from Step A (34.3 g, 119 mmol) in THF (77.6 mL) was added slowly a 1 M solution of BH3-THF in THF (358 mL, 3 eq) and the reaction mixture was stirred for 18 h. After the addition of methanol and stirring for 30 min, purification by column chromatography (silica gel, heptane and MTBE as eluents) afforded the desired product (16.19 g, 49.6%).
• Step C 1-[(3-bromo-5,7-dimethyl-1-adamantyl)methyl]pyrazole [289]
• Step D 5-methyl-1-[(3-bromo-5,7-dimethyl-1-adamantyl)methyl]pyrazole [290] To the solution of the product from Step C (17.88 g, 55.3 mmol) in THF (277 mL) was added butyllithium (2.5 M in THF, 66 mL, 3 eq) at -78°C, then after 1 h, iodomethane (17.2 mL, 5 eq) was added. After 10 min, the reaction mixture was quenched with a saturated solution of NH 4 Cl, extracted with EtOAc and the combined organic layers were dried and concentrated to give the desired product (18.7 g, 100%), which was used in the next step without further purification.
• Step E 2-[[3,5-dimethyl-7-[(5-methylpyrazol-1-yl)methyl]-1-adamantyl]oxy]ethanol [291]
• the mixture of the product from Step D (18.7 g, 55.3 mmol), ethylene glycol (123 mL, 40 eq), and DIPEA (48.2 mL, 5 eq) was stirred at 120°C for 6 h. After the reaction mixture was diluted with water and extracted with EtOAc, the combined organic layers were dried and concentrated to give the desired product (185 g 105%) which was used in the next step without further purification.
• Step F tert-butyl-diphenyl-[2-[[3,5-dimethyl-7-[(5-methylpyrazol-1-yl)methyl]-1- adamantyl] oxy]ethoxy]silane [292]
• Step G tert-butyl-diphenyl-[2-[[3-[(4-iodo-5-methyl-pyrazol-1-yl)methyl]-5,7-dimethyl-1- adamantyl]oxy]ethoxy]silane [293]
• DMF 243 mL
• N-iodosuccinimide 13.6 g, 1.25 eq
• Step H tert-butyl-diphenyl-[2-[[3,5-dimethyl-7-[[5-methyl-4-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)pyrazol-1-yl]methyl]-1-adamantyl]oxy]ethoxy]silane [294] To the product from Step G (17.5 g, 25.6 mmol) in THF (128 mL) was added chloro(isopropyl)magnesium-LiCl (1.3 M in THF, 24 mL, 1.2 eq) at 0°C, stirred for 40 min, treated with 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (15.7 mL, 3 eq), and the reaction mixture was stirred for 10 min.
• Step B (4-methoxyphenyl)methyl 3-bromo-6-[3-(3,6-dichloro-5-methyl-pyridazin-4- yl)propylamino]pyridine-2-carboxylate [296]
• 4-methoxyphenyl)methanol 6.6 mL, 2 eq
• triphenylphosphine 13.9 g, 2 eq
• THF 20 ml
• Step C (4-methoxyphenyl)methyl 3-[1-[[3-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-5,7- dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]-6-[3-(3,6-dichloro-5-methyl- pyridazin-4-yl)propylamino]pyridine-2-carboxylate [297] The mixture of the product of Step B (27.0 g, 50.0 mmol), Preparation 3 (37.5 g, 1.1 eq), Cs 2 CO 3 (48.9 g, 3 eq), Pd(AtaPhos) 2 Cl2 (2.21 g, 0.1 eq) in 1,4-dioxane (300 mL) and H 2 O (50 mL) was kept at 80°C for 6 h.
• Step D (4-methoxyphenyl)methyl 3-[1-[[3-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-5,7- dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]-6-(3-chloro-4-methyl-6,7-dihydro- 5H-pyrido[2,3-c]pyridazin-8-yl)pyridine-2-carboxylate [298] A mixture of the product of Step C (26.0 g, 25.6 mmol) Cs 2 CO 3 (9.87 g, 2 eq), DIPEA (8.9 mL, 2 eq), and Pd(Ataphos) 2 Cl 2 (1.1 g, 0.1 eq) in 1,4-dioxane (128 mL) was stirred in a 200 ml pressure bottle at 110°C for 18 h After dilution with water the mixture was extracted with EtOAc.
• Step E (4-methoxyphenyl)methyl 6-(3-chloro-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl)-3-[1-[[3-(2-hydroxyethoxy)-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylate [299] To the product of Step D (3.40 g, 3.47 mmol) in THF (34.7 mL) was added a 1 M TBAF solution in THF (3.82 mL, 1.1 eq) at 0°C, and the mixture was stirred at RT for 90 min.
• Step F (4-methoxyphenyl)methyl 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7- dihydro-5H-pyrido[2,3-c]pyridazin-8-yl]-3-[1-[[3-(2-hydroxyethoxy)-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylate [300] The mixture of the product of Step E (4.70 g, 6.30 mmol), 1,3-benzothiazol-2-amine (1.90 g, 2 eq), DIPEA (3.30 mL, 3 eq), Pd 2 (dba) 3 (580 mg, 0.1 eq), and XantPhos (730 mg, 0.2
• Step G (4-methoxyphenyl)methyl 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7- dihydro-5H-pyrido[2,3-c]pyridazin-8-yl]-3-[1-[[3,5-dimethyl-7-[2-(p-tolylsulfonyl oxy)ethoxy]-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylate [301] To the product of Step F (3.83 g, 4.48 mmol) and triethylamine (1.87 mL, 3 eq) in DCM (45 mL) was added p-tolylsulfonyl 4-methylbenzenesulfonate (2.19 g, 1.5 eq) at 0°C and the mixture was stirred at RT for 2 h.
• Step H (4-methoxyphenyl)methyl 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7- dihydro-5H-pyrido[2,3-c]pyridazin-8-yl]-3-[1-[[3,5-dimethyl-7-[2-(methylamino) ethoxy]- 1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylate [302] To the product of Step G (2.00 g, 1.98 mmol) in MeCN (9.9 mL) was added a 2 M methanamine solution in THF (9.9 mL, 10 eq) and the mixture was stirred at 50°C for 18 h in a closed bottle.
• Desired product (68.4 g, 104%) was obtained after triturating in heptane-Et2O (3:2).
• 1 H NMR 500 MHz, DMSO-d6) ⁇ ppm 11.96 (br s, 1H), 7.86 (d, 1H), 7.65 (s, 1H), 7.51 (d, 1H), 7.38 (t, 1H), 7.21 (t, 1H), 2.37 (s, 3H); 13 C NMR (125 MHz, DMSO-d6) ⁇ ppm 130.3, 129.5, 126.6, 122.8, 122.3, 17.2;
• HRMS-ESI (m/z): [M+H] + calcd for C 12 H 10 ClN 4 S: 277.0315, found: 277.0305.
• Step B (Z)-N-(6-chloro-4-methyl-pyridazin-3-yl)-3-(2-trimethylsilylethoxymethyl)-1,3- benzothiazol-2-imine [305] To the product of Step A (68.4 g, 247 mmol) and diisopropylethylamine (129 mL, 3 eq) in DCM (1 L) was added 2-(chloromethoxy)ethyl-trimethyl-silane (48.1 mL, 1.1 eq) at 0°C and the mixture was stirred at RT for 15 min.
• Step B methyl 3-bromo-6-(5-chloropentylamino)pyridine-2-carboxylate [307]
• DCM dimethylethyl sulfoxide
• TFA 4.3 mL, 6 eq
• the reaction mixture was stirred at 40°C for 1 h.
• the combined organic phases were dried and concentrated to afford the desired product (3.0 g, 97%) LC/MS (C 12 H 17 BrClN 2 O 2 ) 335 [M+H] + .
• Step C (4-methoxyphenyl)methyl 3-bromo-6-(5-chloropentylamino)pyridine-2- carboxylate [308] After stirring the product of Step B (2.86 g, 8.5 mmol), (4-methoxyphenyl)methanol (1.27 mL, 1.2 eq), La(OiPr) 3 (270 mg, 0.1 eq), 2-(2-hydroxyethoxy)ethanol (0.080 mL, 0.1 eq), and 5A molecular sieves (2.86 g) in hexane (17 mL) at 65 °C for 48 h, the desired product (2.23 g, 59%) was isolated by column chromatography (silica gel, using heptane, EtOAc, as eluents).
• Step D (4-methoxyphenyl)methyl 3-[1-[[3-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-5,7- dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]-6-(5-chloropentylamino) pyridine- 2-carboxylate [309]
• Step E (4-methoxyphenyl)methyl 3-[1-[[3-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-5,7- dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]-6-[5-chloropentyl-[5-methyl-6- [(Z)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino] pyridazin-3- yl]amino]pyridine-2-carboxylate [310] The mixture of the product of Step D (3.23 g, 3.52 mmol), the product of Preparation 6 (2.15 g, 5.28 mmol, 1.5 eq), diisopropylethylamine (1.84 mL, 3 eq), Pd 2 (dba) 3 (322 mg, 0.1 eq), and XantPhos (407 mg, 0.2
• Step F (4-methoxyphenyl)methyl 6-[5-azidopentyl-[5-methyl-6-[(Z)-[3-(2-trimethyl silylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-3-[1-[[3-[2- [tert-butyl(diphenyl)silyl]oxyethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl- pyrazol-4-yl]pyridine-2-carboxylate [311] The mixture of the product of Step E (3.80 g, 3 mmol) and sodium-azide (2.30 g, 12 eq) in 1-methylpyrrolidin-2-one (30 mL) was stirred at 80°C for 18 h.
• Step G (4-methoxyphenyl)methyl 6-[5-azidopentyl-[5-methyl-6-[(Z)-[3-(2-trimethyl silylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-3-[1-[[3-(2- hydroxyethoxy)-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl] pyridine-2- carboxylate [312] To the product of Step F (3.40 g, 2.62 mmol) in THF (27 mL) was added a 1 M solution of TBAF in THF (2.89 mL, 1.1 eq) at 0°C and stirred for 2 h.
• Step H (4-methoxyphenyl)methyl 6-[5-azidopentyl-[5-methyl-6-[(Z)-[3-(2-trimethyl silylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-3-[1-[[3-[2- (dimethylamino)ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carboxylate [313] To the product of Step G (2.34 g, 2.2 mmol) and triethylamine (1.24 mL, 4 eq) in DCM (22 mL) was added p-tolylsulfonyl 4-methylbenzenesulfonate (1.44 g, 2 eq).
• reaction mixture was diluted with brine and extracted with EtOAc.
• the combined organic layers were extracted with 1 M solution of Na2S2O3 and brine, dried, concentrated and purified by flash chromatography (silica gel, using heptane as eluent) to give the desired product (60 g, 80%).
• Step B methyl 2-(tert-butoxycarbonylamino)-5-(3-hydroxyprop-1-ynyl)thiazole-4- carboxylate [315]
• Step C methyl 2-(tert-butoxycarbonylamino)-5-(3-hydroxypropyl)thiazole-4- carboxylate [316]
• the product was purified via flash chromatography column using heptane and EtOAc as eluents to give 31.9 g of the desired product (70.4%).
• Step D methyl 2-(tert-butoxycarbonylamino)-5-[3-(2-fluoro-4-iodo-phenoxy)propyl] thiazole-4-carboxylate [317]
• 2-fluoro-4-iodo-phenol 14 mmol
• 5.00 g of the product from Step C 16 mmol, 1.1 eq
• 4.10 g of PPh 3 1.1 eq
• DIAD 3.10 mL of DIAD (3.20 g, 1.1 eq).
• reaction mixture was directly injected onto a preconditioned silica gel column, and then it was purified via flash chromatography using heptane and EtOAc as eluents.
• the crude product was crystalized from MeOH to give 4.64 g of the desired product (66%).
• Step E methyl 2-[tert-butoxycarbonyl(5-trimethylsilylpent-4-ynyl)amino]-5-[3-(2-fluoro- 4-iodo-phenoxy)propyl]thiazole-4-carboxylate [318] After mixing the product of Step D (5.36 g, 10 mmol, 1 eq) and 5-trimethylsilyl pent-4- yn-1-ol (3.12 g, 20 mmol, 2 eq) in toluene (50 mL) with PPh3 (5.24 g, 2 eq), DIAD (3.9 mL, 2 eq) was added, and stirred at 50°C for 30 min.
• Step F methyl 5-[3-(2-fluoro-4-iodo-phenoxy)propyl]-2-(5-trimethylsilylpent-4- ynylamino)thiazole-4-carboxylate [319] After stirring the product of Step E (6.00 g, 8.9 mmol) in hexafluoroisopropanol (44 mL) at 100°C for 5 hours, volatiles were removed under reduced pressure and the crude product was purified by column chromatography (silica gel, heptane and EtOAc as eluents) to give the desired product (4.46 g, 87%).
• Step G methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluoro- phenoxy]propyl]-2-(5-trimethylsilylpent-4-ynylamino)thiazole-4-carboxylate [320] To the mixture of the product of Step F (4.46 g, 7.7 mmol), Pd(PPh 3 ) 2 Cl 2 (272 mg, 0.05 eq), and CuI (74 mg, 0.05 eq) in diisopropylamine (15 mL) and THF (30 mL) was added tert- butyl N-methyl-N-prop-2-ynyl-carbamate (2.62 g, 2 eq) and stirred at 60°C for 1 h.
• Step H methyl 5-[3-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]-2-fluoro- phenoxy]propyl]-2-[[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzo thiazol-2-ylidene]amino]pyridazin-3-yl]-(5-trimethylsilylpent-4-ynyl)amino]thiazole-4- carboxylate [321] The mixture of the product of Step G (4.44 g, 7.21 mmol), the product of Preparation 6 (3.52 g, 1.2 eq), diisopropylethylamine (3.77 mL, 3 eq), Pd2(dba)3 (660 mg, 0.1 eq), and XantPhos (834 mg, 0.2 eq) in 1,4-dioxane (36
• Step I methyl 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-pent-4-ynyl- amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl] thiazole-4- carboxylate [322]
• the product of Step H (350 mg, 0.35 mmol) in MeCN (3.5 mL) was treated with 70% HF in pyridine (4.57 mL, 100 eq) and stirred at 60°C for 2 h.
• Step J 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-pent-4-ynyl-amino]- 5-[3-[2-fluoro-4-[3-(methylamino)prop-1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid [323]
• the suspention of the product of Step I 160 mg, 0.23 mmol) in THF (2.34 mL) and water (0.47 mL) was treated with LiOH x H 2 O (19 mg, 2 eq), stirred at 60°C for 2 h, and purified by preparative HPLC (Interchim Method) (C18, 25mM aqeous NH 4 HCO 3 , MeCN) to give the desired product (30 mg, 19%).
• Step B methyl 2-[3-(3,6-dichloro-5-methyl-pyridazin-4-yl)propylamino]-5-[3-(2-fluoro-4- iodo-phenoxy)propyl]thiazole-4-carboxylate [325]
• the product from Step A (15.0 g, 20 mmol) in 1,1,1,3,3,3-hexafluoroisopropanol (81 mL) was stirred at 110°C for 18 h. After removing the volatiles under reduced pressure, purification by column chromatography (silica gel, heptane and EtOAc as eluents) afforded the desired product (8.6 g, 66%).
• Step C methyl 2-(3-chloro-4-methyl-6,7-dihydro-5H-pyrido[2,3-c]pyridazin-8-yl)-5-[3-(2- fluoro-4-iodo-phenoxy)propyl]thiazole-4-carboxylate [326]
• a suspension of 3.0 g of the product from Step B (4.69 mmol) and 1.81 g of Cs2CO 3 (2 eq) in 25 mL of 1,4-dioxane were stirred at 80°C for 3 h.
• the product was purified by flash chromatography using DCM-MeOH as eluents to give 2.67 g (94%) of the desired product.
• Step D methyl 2-(3-chloro-4-methyl-6,7-dihydro-5H-pyrido[2,3-c]pyridazin-8-yl)-5-[3-[2- fluoro-4-(2-trimethylsilylethynyl)phenoxy]propyl]thiazole-4-carboxylate [327] To 5.0 g of the product of Step C (8.29 mmol, 1 eq.), 2.34 mL of ethynyl(trimethyl) silane (2 eq), and 10 mL DIPEA in 40 mL of THF were added 182 mg Pd(PPh3) 2 Cl 2 (0.05 eq) and 79 mg CuI (0.05 eq).
• Step E methyl 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[2-fluoro-4-(2-trimethylsilylethynyl) phenoxy] propyl]thiazole-4- carboxylate [328] To 4.25 g of the product from Step D (7.4 mmol, 1.0 eq.), 2.23 g 1,3-benzo thiazol-2- amine (2 eq), and 3.87 mL of DIPEA (3 eq) in 40 mL of cyclohexanol were added 679 mg of Pd 2 (dba) 3 (0.10 eq) and 858 mg of XantPhos (0.20 eq) and the resulting mixture was stirred at 140°C for 30 min.
• Step F 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-(4-ethynyl-2-fluoro-phenoxy)propyl]thiazole-4-carboxylic acid [329]
• Step G 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[4-[3-(4-tert-butoxycarbonylpiperazin-1-yl)prop-1-ynyl]-2-fluoro- phenoxy]propyl]thiazole-4-carboxylic acid [330] The mixture of 200 mg of the product of Step F (0.33 mmol), paraformaldehyde (100 mg), CuI (63 mg) and tert-butyl piperazine-1-carboxylate (620 mg, 10 eq) in EtOH (3.3 mL) was irradiated in an Anton-Paar microwave reactor at 100°C for 1 h.
• Step H 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[2-fluoro-4-(3-piperazin-1-ylprop-1-ynyl)phenoxy]propyl] thiazole- 4-carboxylic acid [331]
• the mixture of the product of Step G (207 mg, 0.25 mmol) and HF x Pyr (10 eq.) in acetonitrile (43 mL) was stirred at 60°C for 25 h
• the product was purified by flash chromatography on silica gel column using DCM and MeOH as eluents to give 143 mg (79%) of the desired product.
• Step A 2-(3-chloro-4-methyl-6,7-dihydro-5H-pyrido[2,3-c]pyridazin-8-yl)-5-[3-(2-fluoro- 4-iodo-phenoxy)propyl]thiazole-4-carboxylic acid
• the mixture of the product of Step C of Preparation 9 (10.0 g, 16 mmol) and LiOH x H 2 O (6.80 g, 10 eq) in THF (81 mL) and water (81 mL) was stirred at 50°C for 6 h. After setting pH to 6 by the addition of HCl, desired product was filtered off (8.20 g, 86%).
• Step B (4-methoxyphenyl)methyl 2-(3-chloro-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl)-5-[3-(2-fluoro-4-iodo-phenoxy)propyl]thiazole-4-carboxylate [334] To the mixture of the product of Step A (8.20 g, 14 mmol), PPh 3 (7.30 g, 2 eq), and (4- methoxyphenyl)methanol (3.80 g, 2 eq) in toluene (70 mL) was added DIAD (5.5 mL, 2 eq). The reaction was stirred at 50°C for 30 min.
• Step C (4-methoxyphenyl)methyl 2-(3-chloro-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl)-5-[3-[2-fluoro-4-(3-piperazin-1-ylprop-1-ynyl)phenoxy]propyl] thiazole- 4-carboxylate [335]
• the mixture of the product of Step B was (7.50 g, 10.6 mmol), 1-prop-2-ynylpiperazine, hydrogen chloride (1:2) (5.00 g, 2.4 eq), Pd(PPh3) 2 Cl 2 (371 mg, 0.05 eq), and CuI (100 mg, 0.05 eq) in THF (52 mL) and DIPA (10 mL) was stirred at 60°C for 1 h.
• Step D (4-methoxyphenyl)methyl 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7- dihydro-5H-pyrido[2,3-c]pyridazin-8-yl]-5-[3-[2-fluoro-4-(3-piperazin-1-ylprop-1- ynyl)phenoxy]propyl]thiazole-4-carboxylate [336] The mixture of the product of Step C (5.30 g, 7.52 mmol), 1,3-benzothiazol-2-amine (3.39 g, 3 eq), DIPEA (3.9 mL, 3 eq), Pd 2 (dba) 3 (344 mg, 0.05 eq) and XantPhos (435 mg, 0.1 eq) in cyclohexanol (37 mL), was stirred at 140°C for 1 h.
• Preparation 14 3,6-dichloro-4-(3-iodopropyl)-5-methyl-pyridazine After stirring PPh 3 (59.3 g, 2 eq), imidazole (15.4 g, 2 eq), and iodine (57.4 g, 2 eq) in 560 mL of DCM for 15 min, 25.0 g of Preparation 1 (113 mmol) was added and stirred for 2 h. The product was purified via flash chromatography using heptane and EtOAc as eluents to give 34.7 g of the desired product (92%).
• Step A methyl 2-(tert-butoxycarbonylamino)-5-[3-[tert-butyl(diphenyl)silyl]oxypropyl] thiazole-4-carboxylate [338] To 77.0 g of the product of Step C of Preparation 8 (243 mmol), imidazole (33.1 g, 2 eq), and DMAP (1.49 g, 0.05 eq) in 970 mL of DMF was added tert-butyl-chloro-diphenyl- silane (93 mL, 1.5 eq) and stirred for 16 h.
• Step B methyl 2-[tert-butoxycarbonyl-[3-(3,6-dichloro-5-methyl-pyridazin-4-yl)propyl] amino]-5-[3-[tert-butyl(diphenyl)silyl]oxypropyl]thiazole-4-carboxylate [339]
• the product of Step A (35.0 g, 63 mmol), Preparation 14 (25.0 g, 1.2 eq), and Cs 2 CO 3 (41.0 g, 2 eq) in 315 mL of acetone were stirred for 1 h.
• Step C methyl 5-[3-[tert-butyl(diphenyl)silyl]oxypropyl]-2-[3-(3,6-dichloro-5-methyl- pyridazin-4-yl)propylamino]thiazole-4-carboxylate [340] After stirring the product of Step B (51.7 g, 60 mmol) in 1,1,1,3,3,3-hexafluoropropan- 2-ol (360 mL) at 100 °C for 18 h, the volatiles were removed under reduced pressure. The crude product was purified via flash chromatography using heptane and EtOAc as eluents to give 36.3 g of the desired product (92%).
• Step D methyl 5-[3-[tert-butyl(diphenyl)silyl]oxypropyl]-2-(3-chloro-4-methyl-6,7- dihydro-5H-pyrido[2,3-c]pyridazin-8-yl)thiazole-4-carboxylate [341] After mixing the product of Step C (36.0 g, 55 mmol) with Cs 2 CO 3 (35.7 g, 2 eq) in 1,4- dioxane (380 mL), the reaction mixture was stirred at 90 °C for 18 h. After diluting the mixture with water, the desired product was collected by filtration (34.0 g, 99%).
• Step E methyl 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[tert-butyl(diphenyl)silyl]oxypropyl]thiazole-4-carboxylate [342] After mixing the product of Step D (6.21 g, 10 mmol), 1,3-benzothiazol-2-amine (3.0 g, 2 eq), and DIPEA (8.7 mL, 2 eq) in cyclohexanol (50 mL), Pd 2 (dba) 3 (915 mg, 0.1 eq) and XantPhos (1.16 g, 0.2 eq) were added, and the reaction mixture was stirred at 140 °C for 1 h.
• Step F methyl 5-[3-[tert-butyl(diphenyl)silyl]oxypropyl]-2-[4-methyl-3-[(Z)-[3-(2- trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]-6,7-dihydro-5H- pyrido[2,3-c]pyridazin-8-yl]thiazole-4-carboxylate [343] After cooling the mixture of the product of Step E (1.64 g, 2.2 mmol), DIPEA (0.77 mL, 2 eq), and DMAP (13 mg, 0.05 eq) in DCM (12 mL) to -20 °C, 2-(chloromethoxy)ethyl-trimethyl- silane (0.61 mL, 1.55 eq) was added, and the reaction mixture was stirred for 18 h.
• Step G methyl 5-(3-hydroxypropyl)-2-[4-methyl-3-[(Z)-[3-(2- trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]-6,7-dihydro-5H- pyrido[2,3-c]pyridazin-8-yl]thiazole-4-carboxylate [344]
• the product of Step F (1.56 g, 1.8 mmol) and a 1 M THF solution of TBAF (2.1 mL, 1.2 eq) were stirred in THF (18 mL) for 4 h.
• Step H methyl 5-(3-iodopropyl)-2-[4-methyl-3-[(Z)-[3-(2-trimethylsilylethoxymethyl)-1,3- benzothiazol-2-ylidene]amino]-6,7-dihydro-5H-pyrido[2,3-c]pyridazin-8-yl]thiazole-4- carboxylate [345] After stirring PPh 3 (594 mg, 1.1 eq), imidazole (154 mg, 1.1 eq), and iodine (574 mg, 1.1 eq) in DCM (10 mL) for 30 min, the product of Step G (1.29 g, 2 mmol) in 1 mL of DCM was added at 0 °C.
• Step I methyl 5-[3-(methylamino)propyl]-2-[4-methyl-3-[(Z)-[3-(2-trimethylsilylethoxy methyl)-1,3-benzothiazol-2-ylidene]amino]-6,7-dihydro-5H-pyrido[2,3-c]pyridazin-8- yl]thiazole-4-carboxylate [346] To the product of Step H (850 mg, 1.1 mmol) in NMP (1.0 mL) and MeCN (11 mL) was added 4.0 mL of a 2 M solution of methanamine in THF (7 eq), and the reaction mixture was stirred at 50 °C for 3 h.
• Preparation 16 6-[5-azidopentyl-[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3- yl]amino]-3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [347] To Preparation 7 (100 mg, 0.092 mmol) in acetonitrile (1.9 mL) was added pyridine, hydrogen fluoride (1:1) (25 eq) and the reaction was stirred at 60 °C for 4 h.
• the reaction mixture was diluted with saturated brine, then it was extracted with EtOAc. The combined organic layers were extracted with 1 M Na 2 S 2 O 3 , then with brine again. Then dried over Na 2 SO 4 , filtered and the filtrate was concentrated under reduced pressure.
• the crude product was purified via flash column chromatography using heptane as eluent to obtain 60 g (156 mmol, 80%) of the desired product.
• Step B methyl 2-(tert-butoxycarbonylamino)-5-(3-hydroxyprop-1-ynyl)thiazole-4- carboxylate [350] A 500 mL oven-dried, one-necked, round-bottom flask was equipped with a PTFE- coated magnetic stirring bar and fitted with a reflux condenser.
• Step C methyl 2-(tert-butoxycarbonylamino)-5-(3-hydroxypropyl)thiazole-4- carboxylate [351]
• An 1 L oven-dried pressure bottle equipped with a PTFE-coated magnetic stirring bar was charged with 44.75 g of the product from Step B (143.3 mmol, 1 eq.), 7.62 g of Pd/C (7.17 mmol, 0.05 eq.) in 340 mL of ethanol, and then placed under a nitrogen atmosphere using hydrogenation system. After that it was filled with 4 bar H 2 gas and stirred at rt overnight. Full conversion was observed, but only the olefin product was formed.
• Step D methyl 2- ⁇ [(tert-butoxy)carbonyl]amino ⁇ -5-[3-(2-fluoro-4-iodophenoxy)propyl]- 1,3-thiazole-4-carboxylate
• Step E methyl 2-(tert-butoxycarbonylamino)-5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2- fluoro-phenoxy]propyl]thiazole-4-carboxylate [353]
• a 250 mL oven-dried, one-necked, round-bottom flask was equipped with a PTFE- coated magnetic stirring bar and fitted with a reflux condenser.
• Step D 10 mmol, 1 eq.
• N,N-dimethylprop-2-yn-1-amine 20 mmol, 2 eq.
• DIPA 142.7 mmol, 14.27 eq.
• 50 mL of dry THF were added and the system was flushed with argon.
• 220 mg of Pd(PPh3) 2 Cl2 0.5 mmol, 0.05 eq.
• 95 mg of CuI 0.5 mmol, 0.05 eq.
• Step F methyl 2-[tert-butoxycarbonyl-[4-(tert-butoxycarbonylamino)butyl]amino]-5-[3- [4-[3-(dimethylamino)prop-1-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate [354] Using Mitsunobu General Procedure starting from 250 mg of Step E (0.51 mmol, 1 eq.) and 193 mg of tert-butyl N-(4-hydroxybutyl)carbamate (1.02 mmol, 2 eq.) as the appropriate alcohol, 220 mg (65%) of the desired product were obtained.
• Step G methyl 2-[4-(tert-butoxycarbonylamino)butylamino]-5-[3-[4-[3- (dimethylamino)prop-1-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate [355] Using Deprotection with HFIP General Procedure starting from 215 mg of the product from Step F (0.33 mmol, 1 eq.) as the appropriate Boc protected amine, 137 mg (75%) of the desired product were obtained.
• Step H methyl 2-[4-(tert-butoxycarbonylamino)butyl-[5-methyl-6-[(Z)-[3-(2- trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-5- [3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylate [356] Using Buchwald General Procedure II starting from 133 mg of the product from Step G (0.24 mmol, 1 eq.) and 120 mg of Preparation 6 (0.29 mmol, 1.25 eq.) as the appropriate halide, 220 mg (98%) of the desired product were obtained.
• Step I 2-[4-aminobutyl-[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]amino]- 5-[3-[4-[3-(dimethylamino)prop-1-ynyl]-2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid [357] Using Deprotection and Hydrolysis General Procedure followed by repurification via reverse phase preparative chromatography (C18, 0.1% TFA in water : MeCN) starting from the product from Step H, the TFA-salt of the desired product was obtained.
• Step B methyl 6-[tert-butoxycarbonyl-[4-[tert- butoxycarbonyl(methyl)amino]butyl]amino]-3-[1-[[3-[2-[tert- butyl(diphenyl)silyl]oxyethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carboxylate [359] The mixture of 7.7 g (14.9 mmol) of the product of Step A and 10.2 g (1 eq) of Preparation 3 in a mixture of 90 mL of 1,4-dioxane and 15 mL of water was treated with 14.6 (3 eq) of Cs 2 CO 3 and 0.66 g (0.1 eq) of Pd(AtaPhos) 2 Cl 2 .
• Step C methyl 6-[tert-butoxycarbonyl-[4-[tert- butoxycarbonyl(methyl)amino]butyl]amino]-3-[1-[[3-(2-hydroxyethoxy)-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylate [360]
• Step D methyl 6-[tert-butoxycarbonyl-[4-[tert- butoxycarbonyl(methyl)amino]butyl]amino]-3-[1-[[3,5-dimethyl-7-[2-(p- tolylsulfonyloxy)ethoxy]-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2- carboxylate [361]
• the mixture of 3.6 g (4.78 mmol) of the product of Step C and 2 mL (3 eq) of triethylamine in 62 mL of DCM was treated with 2.34 g (1.5 eq) of p-tolylsulfonyl 4- methylbenzenesulfonate at 0 °C.
• Step E methyl 6-[tert-butoxycarbonyl-[4-[tert- butoxycarbonyl(methyl)amino]butyl]amino]-3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7- dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylate [362] After the treatment of 3.7 g (4.07 mmol) of the product of Step D with 20.37 mL (10 eq) of a 2 M solution of dimethylamine in MeOH at 50 °C for 2 h, the mixture was diluted with 10% aqueous K 2 CO 3 solution and extracted with DCM.
• Step F methyl 6-[4-[tert-butoxycarbonyl(methyl)amino]butylamino]-3-[1-[[3-[2- (dimethylamino)ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carboxylate [363]
• the product from Step E (3.17 g, 4.06 mmol) in 1,1,1,3,3,3-hexafluoroisopropanol (24 mL) was stirred at 110 °C for 18 h.
• Step G methyl 6-[4-[tert-butoxycarbonyl(methyl)amino]butyl-[5-methyl-6-[(Z)-[3-(2- trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]-3- [1-[[3-[2-(dimethylamino)ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carboxylate [364] The mixture of the product of Step F (700 mg, 1.03 mmol), the product of Preparation 6 (711.3 mg, 1.7 eq), diisopropylethylamine (0.54 mL, 3 eq), Cs 2 CO 3 (1.0 g, 3 eq), Pd 2 (dba) 3 (94 mg, 0.1 eq), and XantPhos (119 mg,
• Step H 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4- (methylamino)butyl] amino]-3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [365] The product of Step G (1100 mg, 0.84 mmol) in 1,4-dioxane (4 mL) was treated with a 1 M solution of hydrogen chloride in 1,4 dioxane (40 eq) for 1 h.
• Step I (4-methoxyphenyl)methyl 6-[[6-[(Z)-3H-1,3-benzothiazol-2-ylideneamino]-5- methyl-pyridazin-3-yl]-[4-(methylamino)butyl]amino]-3-[1-[[3-[2- (dimethylamino)ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carboxylate [366] The product of Step H (590 mg, 0.72 mmol) and (4-methoxyphenyl)methanol (267 uL, 3.0 eq) were suspended in dry toluene (15 mL), and then tetraethoxytitanium (30 uL, 0.2 eq) was added.
• Step G To the product of Preparation 4, Step G (500 mg) in acetonitrile (5 mL) was added pyrrolidine (6.5 eq) and the reaction mixture was stirred at 50 °C for 18 h. After treatment of the reaction with KOH (3.6 eq), the mixture was stirred at 50 °C for 2 h. The product was purified by preparative HPLC (using acetonitrile and 5mM aqueous NH 4 HCO 3 solution as eluents) to give the desired product.
• HRMS-ESI (m/z): [M+H] + calcd for C 44 H 54 N 9 O 3 S: 788,4064, found: 788.4068.
• Step B N-[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]piperidine-4-carboxamide [370]
• the mixture of the product of Step A (0.099 mmol) and a 4 M hydrogen chloride solution in 1,4-dioxane (10 eq) in 5 mL of 1,4-dioxane was stirred for 18 h. The volatiles were removed under reduced pressure and it gave 67 mg of the desired product.
• the crude product was purified by column chromatography (silica gel, heptane and EtOAc as eluents) or preparative chromatography (using acetonitrile and 25 mM aqueous TFA solution as eluents) to give the desired product.
• bifunctional Bcl-xL degrader compounds comprising a Bcl-xL payload covalently linked to a degrading compound (DSM), were synthesized using exemplary methods described in this example.
• DSM degrading compound
• General Procedures General procedure for production of VHL ligand-based degraders via alkylation [415] To the product of Preparation 4 and DIPEA (1.0 mL/mmol) in a mixture of MeCN (10 mL/mmol) and 1-methyl-2-pyrrolidinone (10 mL/mmol) was added the appropriate alkylating agent (1.5 eq) and the mixture was stirred at 70°C until appropriate conversion was achieved.
• Example 1 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[16-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methyl thiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl] amino]-16-oxo-hexadecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl] methyl] - 5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid Step A: (2S,4R)-4-hydroxy-1-[(2S)-2-(16-hydroxyhexadecanoylamin
• Step B [16-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl] ethyl] carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-16-oxo-hexadecyl]4- methylbenzenesulfonate [419] Using the General procedure for the tosylation of the hydroxyalkyl VHL ligand- derivatives starting from the product of Step A (90 mg), 49 mg of the desired product was obtained.
• Step C 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[16-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl] ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-16-oxo-hexadecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [420] Using the General procedure for production of VHL ligand-based degraders via alkylation starting from the product of Prepar
• Step B [12-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl] ethyl] carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-12-oxo-dodecyl]4- methyl benzenesulfonate [422] Using the General procedure for the tosylation of the hydroxyalkyl VHL ligand- derivatives starting from the product of Step A (110 mg), 100 mg of the desired product was obtained.
• Step C 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[12-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl] ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-12-oxo-dodecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [423] Using the General procedure for production of VHL ligand-based degraders via alkylation starting from the product of Prepar
• Step A (2S,4R)-1-[(2S)-2-(7-bromoheptanoylamino)-3,3-dimethyl-butanoyl]-4-hydroxy- N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide
• (2S,4R)-1-[(2S)- 2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] pyrrolidine-2-carboxamide hydrogen chloride (1:1) (0.42 mmol) and 7-bromoheptanoic acid as the appropriate acid, 181 mg of the desired product was obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[7-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl] ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-7-oxo-heptyl]-methyl -amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]- 5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [425] Using the General procedure for production of VHL ligand-based degraders via alkylation starting from the product of Preparation 4
• Step B [14-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl] ethyl] carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-14-oxo-tetradecyl]4- methylbenzenesulfonate [427] Using the General procedure for the tosylation of the hydroxyalkyl VHL ligand- derivatives starting from the product of Step A (100 mg), 105 mg of the desired product was obtained.
• Step C 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[14-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl] ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-14-oxo-tetradecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [428] Using the General procedure for production of VHL ligand-based degraders via alkylation starting from the product of Prepar
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[9-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-9-oxo-nonyl]-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]- 5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [430] Using the General procedure for production of VHL ligand-based degraders via alkylation starting from the product of Preparation 4 (30 mg)
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[8-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-8-oxo-octyl]-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [432] Using the General procedure for production of VHL ligand-based degraders via alkylation starting from the product of Preparation 4
• Step B [15-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-15-oxo-pentadecyl]4- methylbenzenesulfonate [434] Using the General procedure for the tosylation of the hydroxyalkyl VHL ligand- derivatives starting from the product of Step A (150 mg), 53 mg of the desired product was obtained.
• Step C 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[15-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl] ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-15-oxo-pentadecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [435] Using the General procedure for production of VHL ligand-based degraders via alkylation starting from the product of Pre
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[6-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl] ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-6-oxo-hexyl]-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]- 5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [437] Using the General procedure for production of VHL ligand-based degraders via alkylation starting from the product of Preparation 4 (30
• Step B 2-(2,6-dioxo-3-piperidyl)-4-[2-(2-iodoethoxy)ethylamino]isoindoline-1,3-dione [439] Using the General procedure for the iodination of hydroxyalkyl derivative of thalidomide starting from the product of Step A (60 mg), 55 mg of the desired product was obtained.
• Step C 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[2-[[2-[[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4- yl]amino]ethoxy] ethyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [440] Using the General procedure for the production of thalidomide-based degraders via alkylation starting from the product of Preparation 4 (30 mg) and the product of Step B as the appropriate alkylating agent, 27 mg of the desired product was obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[8-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5- yl]oxyoctyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol- 4-yl]pyridine-2-carboxylic acid [442] Using the General procedure for the production of thalidomide-based degraders via alkylation starting from the product of Preparation 4 (20 mg) and the product of Step A as the appropriate alkylating agent, 17 mg of the desired product was obtained.
• Step B 2-(2,6-dioxo-3-piperidyl)-4-[2-[2-[2-(2-iodoethoxy)ethoxy]ethoxy] ethylamino] isoindoline-1,3-dione [444]
• Step C 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[2-[2-[2-[2-[[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4- yl]amino] ethoxy]ethoxy]ethoxy]ethyl-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [445] Using the General procedure for the production of thalidomide-based degraders via alkylation starting from the product of Preparation 4 (40 mg) and the product of Step B as the appropriate alkylating agent, 7 mg of the desired product was obtained.
• Step B 2-(2,6-dioxo-3-piperidyl)-4-[2-[2-(2-iodoethoxy)ethoxy]ethylamino] isoindoline- 1,3-dione [447] Using the General procedure for the iodination of hydroxyalkyl derivative of thalidomide starting from the product of Step A (40 mg), 34 mg of the desired product was obtained.
• Step C 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[2-[2-[2-[[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4- yl]amino]ethoxy] ethoxy]ethyl-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [448] Using the General procedure for the production of thalidomide-based degraders via alkylation starting from the product of Preparation 4 (50 mg) and the product of Step B as the appropriate alkylating agent, 12 mg of the desired product was obtained.
• Step B 2-(2,6-dioxo-3-piperidyl)-4-[2-[2-[2-[2-(2-iodoethoxy)ethoxy]ethoxy] ethoxy]ethylamino]isoindoline-1,3-dione [450] Using the General procedure for the iodination of hydroxyalkyl derivative of thalidomide starting from the product of Step A (500 mg), 272 mg of the desired product was obtained.
• Step C 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[2-[2-[2-[2-[2-[2-[[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo- isoindolin-4-yl]amino] ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethyl-methyl- amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2- carboxylic acid [451] Using the General procedure for the production of thalidomide-based degraders via alkylation starting from the product of Preparation 4 (30 mg) and the product of Step B as the appropriate alkylating agent, 22 mg of the desired product
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[6-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5- yl]oxyhexyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol- 4-yl]pyridine-2-carboxylic acid [453] Using the General procedure for the production of thalidomide-based degraders via alkylation starting from the product of Preparation 4 (15 mg) and the product of Step A as the appropriate alkylating agent, 12 mg of the desired product was obtained.
• Step B 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[2-fluoro-4-[3-[4-[7-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[4-(4- methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-7-oxo-heptanoyl]piperazin-1-yl]prop-1-ynyl]phenoxy]propyl] thiazole-4- carboxylic acid [455] Using the General procedure for production of degraders via acylation starting from Preparation 9 (20 mg) and the product of Step A as the appropriate acid, the desired product was obtained (20 mg).
• Step B 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[2-fluoro-4-[3-[4-[8-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-8-oxo-octanoyl]piperazin-1-yl]prop-1-ynyl]phenoxy]propyl]thiazole-4- carboxylic acid [457] Using the General procedure for production of degraders via acylation starting from Preparation 9 (80 mg) and the product of Step A as the appropriate acid, the desired product was obtained
• Step B 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[2-fluoro-4-[3-[4-[7-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-7-oxo-heptanoyl]piperazin-1-yl]prop-1-ynyl]phenoxy]propyl] thiazole-4- carboxylic acid [459] Using the General procedure for production of degraders via acylation starting from Preparation 9 (80 mg) and the product of Step A as the appropriate acid, the desired product was obtained
• Step B 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[2-fluoro-4-[3-[4-[5-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[4-(4- methylthiazol-5-yl)phenyl] methylcarbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-5-oxo-pentanoyl] piperazin-1-yl]prop-1-ynyl]phenoxy]propyl] thiazole-4- carboxylic acid [461] Using the General procedure for production of degraders via acylation starting from Preparation 9 (37 mg) and the product of Step A as the appropriate acid, the desired product was obtained (10 mg).
• Step B 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[2-fluoro-4-[3-[4-[6-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[4-(4- methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-6-oxo-hexanoyl]piperazin-1-yl]prop-1-ynyl]phenoxy]propyl] thiazole-4- carboxylic acid [463] Using the General procedure for production of degraders via acylation starting from Preparation 9 (50 mg) and the product of Step A as the appropriate acid, the desired product was obtained (45 mg).
• Step B 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[2-fluoro-4-[3-[4-[8-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[4-(4- methylthiazol-5-yl)phenyl] methylcarbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-8-oxo-octanoyl] piperazin-1-yl]prop-1-ynyl]phenoxy]propyl]thiazole-4- carboxylic acid [465] Using the General procedure for production of degraders via acylation starting from Preparation 9 (25 mg) and the product of Step A as the appropriate acid, the desired product was obtained (15 mg).
• Step B 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[2-fluoro-4-[3-[4-[4-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[4-(4- methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-4-oxo-butanoyl]piperazin-1-yl]prop-1-ynyl]phenoxy]propyl]thiazole-4- carboxylic acid [467] Using the General procedure for production of degraders via acylation starting from Preparation 9 (40 mg) and the product of Step A as the appropriate
• Example 25 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3-c] pyridazin-8-yl]-5-[3-[4-[3-[4-[8-[2-[2-[[2-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4- yl]oxyacetyl]amino]ethoxy]ethylamino]-8-oxo-octanoyl]piperazin-1-yl]prop-1-ynyl]-2- fluoro-phenoxy]propyl]thiazole-4-carboxylic acid Step A: 8-[2-[2-[[2-[2-[2-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]oxyacetyl]
• Step B 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[4-[3-[4-[8-[2-[2-[[2-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin- 4-yl]oxyacetyl]amino]ethoxy]ethylamino]-8-oxo-octanoyl]piperazin-1-yl]prop-1-ynyl]-2- fluoro-phenoxy]propyl]thiazole-4-carboxylic acid [472] Using the General procedure for production of degraders via acylation starting from Preparation 9 (50 mg) and the product of Step A as the appropriate acid, the desired product was obtained (16 mg).
• Step B 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[4-[3-[4-[5-[2-[2-[[2-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin- 4-yl]oxyacetyl] amino]ethoxy]ethylamino]-5-oxo-pentanoyl]piperazin-1-yl]prop-1-ynyl]- 2-fluoro-phenoxy]propyl]thiazole-4-carboxylic acid [474] Using the General procedure for production of degraders via acylation starting from Preparation 9 (50 mg) and the product of Step A as the appropriate acid, the desired product was obtained (18 mg).
• Step B 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[3-[1-[2-[2-[2-[[2- [2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]oxyacetyl]amino]ethoxy] ethoxy]ethoxy] ethyl]triazol-4-yl]propyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino) prop- 1-ynyl]phenoxy] propyl]thiazole-4-carboxylic acid [476] To a solution of CuSO 4 x 5 H 2 O (9.87 mg, 0.056 mmol) in water (1.2 mL) was added THTPA (24.13 mg, 0.056 mmol).
• Step A tert-butyl 2-[2-(2-prop-2-ynoxyethoxy)ethoxy]acetate
• 2-(2-prop-2-ynoxyethoxy)ethanol (1 g, 6.94 mmol) in THF (30ml)
• sodium hydride 60% in mineral oil (282 mg, 7.08 mmol) at 0°C
• a solution of tert-butyl 2-bromoacetate (1.54 mL, 10.4 mmol) in THF (5ml) was added and the reaction was stirred at RT for 2 h.
• the reaction was quenched with water and extracted with ethyl acetate.
• Step B 2-[2-(2-prop-2-ynoxyethoxy)ethoxy]acetic acid [478]
• the product of Step A (785 mg, 3.04 mmol) in DCM was treated with a 4N solution of HCl in dioxane (3.79 mL, 15.2 mmol). The reaction was stirred for 4 h and additional HCl was added (3.79 mL, 15.2 mmol). After 16 h stirring, the reaction was concentrated to give the desired product (470 mg).
• Step C N-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindolin-4-yl]-2-[2-(2-prop-2-ynoxy ethoxy) ethoxy]acetamide
• 3-(4-amino-1-oxo-isoindolin-2-yl)piperidine-2,6-dione (0.137 mL, 0,77 mmol) and the product of Step B (155.98 mg, 0.771 mmol) in DMSO (8ml) were successively added HATU (293.3 mg, 0.77 mmol), HOAt (105.0 mg, 0.77 mmol), and DIEA (0.686 mL, 3.85 mmol) and the mixture was stirred for 1 h.
• Step D (4-methoxyphenyl)methyl 3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]-6-[5-[4-[2-[2-[[2-(2,6-dioxo-3-piperidyl)-1- oxo-isoindolin-4-yl]amino]-2-oxo-ethoxy]ethoxy]ethoxymethyl]triazol-1-yl]pentyl-[5- methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2- ylidene]amino]pyridazin-3-yl]amino]pyridine-2-carboxylate [480] To CuSO45H 2 O (8.19 mg, 46.1 ⁇ mol) in water (0.7 mL) was added Na-L-(2-
• Step E 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[4-[2-[2-[[2- (2,6-dioxo-3-piperidyl)-1-oxo-isoindolin-4-yl]amino]-2-oxo-ethoxy]ethoxy] ethoxymethyl] triazol-1-yl]pentyl]amino]-3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7- dimethyl-1-adamantyl] methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [481] To the product of Step D (30 mg, 19.6 ⁇ mol) in acetonitrile (0.1 mL) was added pyridine, hydrogen fluoride (1:1) (89 ⁇ L, 982 ⁇ mol) and the mixture was stirred at 60 °
• Example 29 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[3-[1-[2-[2-[2- [2-[2-[3-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[4-(4-methylthiazol-5-yl)phenyl]methyl carbamoyl] pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-3-oxo- propoxy]ethoxy]ethoxy]ethoxy] ethoxy]ethoxy]triazol-4-yl]propyl]amino]-5-[3-[2-fluoro- 4-[3-(methylamino)prop-1-ynyl] phenoxy]propyl]thiazole-4-carboxylic acid Step A: (2S,4R)-1-[(2S)-2-[3-[2-[2-[2-[2-[2-
• Step B 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[3-[1-[2-[2-[2-[2-[2-[2-[2-[2- [3-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[4-(4-methylthiazol-5-yl)phenyl]methyl carbamoyl] pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-3-oxo-propoxy] ethoxy]ethoxy] ethoxy] ethoxy]ethoxy]ethyl]triazol-4-yl]propyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop- 1-ynyl]phenoxy]propyl]thiazole-4-carboxylic acid [483] To CuSO4 x 5 H 2 O (9.86 mg,
• Step B 2-[[6-[(Z)-3H-1,3-benzothiazol-2-ylideneamino]-5-methyl-pyridazin-3-yl]-[3-[1- [8-[[2-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]oxyacetyl]amino]octyl] triazol- 4-yl] propyl]amino]-5-[3-[2-fluoro-4-[3-(methylamino)prop-1-ynyl]phenoxy] propyl]thiazole-4-carboxylic acid [485] To a solution of CuSO 4 x 5 H 2 O (4.8 mg, 0.027 mmol) in water (1.5 mL) was added THTPA (13.25 mg, 0.030 mmol).
• Example 31 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3-c] pyridazin-8-yl]-5-[3-[2-fluoro-4-[3-[4-[12-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-12-oxo-dodecyl]piperazin-1-yl]prop-1-ynyl]phenoxy]propyl] thiazole-4- carboxylic acid [486] After stirring the product of Preparation 11 (15 mg, 0.018 mmol), the product of Step B of Example 2 (23 mg, 1.6 eq), and DIPEA
• Example 32 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl- amino]-5-[3-[4-[3-[[2-[2-[[2-[2-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4- yl]oxyacetyl]amino]ethoxy]ethylamino]-2-oxo-ethyl]-methyl-amino]prop-1-ynyl]-2- fluoro-phenoxy]propyl]thiazole-4-carboxylic acid
• Step A ethyl 5-(3-chloropropyl)-2-(methylamino)thiazole-4-carboxylate [487] To the suspension of 2.25 g of methylthiourea (25.0 mmol, 1 eq.) in 100 mL of ethanol was added dropwise 7.46 g of ethyl 3-bromo-6-chloro-2-oxo-hexanoate (1.1 eq.) at 0 °C. After 15 min stirring at 0 °C, 7 mL of TEA (5.06 g, 2 eq.) was added. After stirring at RT for 18 h and concentration, the residue was portioned between EtOAc and water.
• Step B ethyl 5-(3-chloropropyl)-2-[methyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilyl ethoxy methyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate
• Step C ethyl 5-(3-iodopropyl)-2-[methyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxy methyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4-carboxylate [489]
• the mixture of 2.6 g of the product from Step B (4.1 mmol, 1 eq.), 1.23 g of NaI (2 eq.) in 20 mL of acetone was stirred at 60 °C for 3 days.
• Step D ethyl 5-[3-(2-fluoro-4-iodo-phenoxy)propyl]-2-[methyl-[5-methyl-6-[(Z)-[3-(2- trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino ]thiazole-4-carboxylate [490] To the mixture of the product of Step C (3.62 g, 5.0 mmol) and Cs 2 CO 3 (3.25 g, 2 eq) in acetone (25 mL) was added 2-fluoro-4-iodo-phenol (1.20 g, 1 eq). The reaction was stirred for 3 h.
• Step E 5-[3-(2-fluoro-4-iodo-phenoxy)propyl]-2-[methyl-[5-methyl-6-[(Z)-[3-(2-trimethyl silylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4- carboxylic acid [491]
• the mixture of the product of Step D (2.0 g, 2.4 mmol) and LiOH*H 2 O (1.0 g, 10 eq) was stirred in a mixture of 1,4-dioxane (10 mL), EtOH (75 mL), and water (22 mL) at 80 °C for 30 min.
• Step F (4-methoxyphenyl)methyl 5-[3-(2-fluoro-4-iodo-phenoxy)propyl]-2-[methyl-[5- methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino] pyridazin-3-yl]amino]thiazole-4-carboxylate [492] To the mixture of the product of Step E (1.49 g, 1.84 mmol) and Cs2CO 3 (1.20 g, 2 eq) in DMF (10 ml) was added 1-(chloromethyl)-4-methoxy-benzene (0.28 mL, 1.1 eq).
• reaction wasstirred at 70 °C for 18 h. After dilution with water, the mixture was extracted with EtOAc. The organic phases were dried, concentrated and purified by flash column chromatography column using heptane and EtOAc as eluents to give 612 mg of the desired product.
• Step G (4-methoxyphenyl)methyl 5-[3-[4-[3-[[2-[2-[[2-[2-[2-[2-[2-[2-[2-[2-[2-(2,6-dioxo-3-piperidyl)-1,3- dioxo-isoindolin-4-yl]oxyacetyl]amino]ethoxy]ethylamino]-2-oxo-ethyl]-methyl-amino] prop-1-ynyl]-2-fluoro-phenoxy]propyl]-2-[methyl-[5-methyl-6-[(E)-[3-(2-trimethylsilyl ethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3-yl]amino]thiazole-4- carboxylate [493] To the mixture of the product of Step F (215 mg, 0.23 mmol), Pd(PPh 3 ) 2 Cl 2 (33 mg,
• Step H 2-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-methyl-amino]-5-[3- [4-[3-[[2-[2-[[2-[2-[2-[2-[2-[2-(2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]oxyacetyl] amino] ethoxy]ethylamino]-2-oxo-ethyl]-methyl-amino]prop-1-ynyl]-2-fluoro-phenoxy]propyl] thiazole-4-carboxylic acid [494] After stirring the product of Step G (187 mg, 0.14 mmol) in MeCN (5 mL)
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[10-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5- yl]oxydecyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol- 4-yl]pyridine-2-carboxylic acid [525] Using the General procedure for the production of thalidomide-based degraders via alkylation starting from the product of Preparation 4 (35 mg) and the product of Step A as the appropriate alkylating agent, 9 mg of the desired product was obtained.
• Example 62 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[4-[2-[2-[2- [[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4- yl]amino]ethoxy]ethoxy]ethoxymethyl] triazol-1-yl]pentyl]amino]-3-[1-[[3-[2- (dimethylamino)ethoxy]-5,7-dimethyl-1-adamantyl] methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carboxylic acid [527] The product was synthesized according to the procedure described for Example 61, using 2-(2,6-dioxo-3-piperidyl)-4-[2-[2-(2-prop-2-ynoxyethoxy)ethoxy]ethylamino
• Step B (4-methoxyphenyl)methyl 3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]-6-[5-[4-[2-[2-[[2-[2-[2-[2-[2-(2,6-dioxo-3-piperidyl)- 1,3-dioxo-isoindolin-4-yl]oxyacetyl]amino]ethoxy]ethoxy]ethoxymethyl]triazol-1- yl]pentyl-[5-methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2- ylidene]amino] pyridazin-3-yl]amino]pyridine-2-carboxylate [529] The product was synthesized according to the procedure described for Step D of Example 28, using Step A as
• Step C 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[4-[2-[2-[[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2- (2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4- yl]oxyacetyl]amino]ethoxy]ethoxy]ethoxy methyl]triazol-1-yl]pentyl]amino]-3-[1-[[3-[2- (dimethylamino)ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carboxylic acid [530] The product was synthesized according to the procedure described for Step E of Example 28, using Step B as a starting material (56%).
• Step B (4-methoxyphenyl)methyl 3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]-6-[5-[4-[7-[[2-[2-(2,6-dioxo-3-piperidyl)-1,3- dioxo-isoindolin-4-yl]oxyacetyl]amino]heptyl]triazol-1-yl]pentyl-[5-methyl-6-[(Z)-[3-(2- trimethylsilylethoxymethyl)-1,3-benzothiazol-2-ylidene]amino]pyridazin-3- yl]amino]pyridine-2-carboxylate [532]
• the product was synthesized according to the procedure described for Step D of Example 28, using Step A as an appropriate acetylene (62%).
• Step C 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[4-[7-[[2-[2-(2,6- dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]oxyacetyl]amino]heptyl]triazol-1- yl]pentyl]amino]-3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7-dimethyl-1-adamantyl]methyl]- 5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [533] The product was synthesized according to the procedure described for Step E of Example 28, using Step B as a starting material (51%).
• Step B (4-methoxyphenyl)methyl 3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]-6-[5-[4-[2-[[2-[2-[2-(2,6-dioxo-3-piperidyl)-1,3- dioxo-isoindolin-4-yl]oxyacetyl]amino]ethoxy]ethoxymethyl]triazol-1-yl]pentyl-[5- methyl-6-[(Z)-[3-(2-trimethylsilylethoxymethyl)-1,3-benzothiazol-2- ylidene]amino]pyridazin-3-yl]amino]pyridine-2-carboxylate [535] The product was synthesized according to the procedure described for Step D of Example 28, using Step A as an appropriate acetylene (53%).
• Step C 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[5-[4-[2-[2-[[2-[2- (2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4- yl]oxyacetyl]amino]ethoxy]ethoxymethyl]triazol-1-yl]pentyl]amino]-3-[1-[[3-[2- (dimethylamino)ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carboxylic acid [536] The product was synthesized according to the procedure described for Step E of Example 28, using Step B as a starting material (56%).
• Step B methyl 5-[3-[[5-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-5-oxo- pentyl]-methyl-amino]propyl]-2-[4-methyl-3-[(Z)-[3-(2-trimethylsilylethoxymethyl)-1,3- benzothiazol-2-ylidene]amino]-6,7-dihydro-5H-pyrido[2,3-c]pyridazin-8-yl]thiazole-4- carboxylate [538] The mixture of 75 mg of Preparation 15 (0.11 mmol), the product of Step A (85 mg, 1.2 eq), DIPEA (0.11 mL) in MeCN
• Step C 2-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-5-[3-[[5-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-5-oxo- pentyl]-methyl-amino]propyl]thiazole-4-carboxylic acid [539] After stirring the product of Step B (40 mg, 0.034 mmol) with LiOH x H 2 O (14 mg, 10 eq) in THF (0.17 ml) and water (0.17 ml) at 50 °C for 5 h, concentrated HC
• Step A 3-[5-[1-(6-bromohexanoyl)-4-piperidyl]-1-oxo-isoindolin-2-yl]piperidine-2,6- dione [540] Using General procedure for the acylation of piperidinyl-isoindolinone starting from 3- [1-oxo-5-(4-piperidyl)isoindolin-2-yl]piperidine-2,6-dione, hydrochloride (1:1) (0.42 mmol) and 6-bromohexanoic acid as the appropriate acid, 165 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[6-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindolin-5-yl]-1- piperidyl]-6-oxo-hexyl]-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [541] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and the product of Step A as the appropriate alkylating agent, 9 mg of the desired product were obtained.
• Step A 3-[5-[1-(8-bromooctanoyl)-4-piperidyl]-1-oxo-isoindolin-2-yl]piperidine-2,6- dione [542] Using General procedure for the acylation of piperidinyl-isoindolinone starting from 3- [1-oxo-5-(4-piperidyl)isoindolin-2-yl]piperidine-2,6-dione, hydrochloride (1:1) (0.42 mmol) and 8-bromooctanoic acid as the appropriate acid, 220 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[8-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindolin-5-yl]-1- piperidyl]-8-oxo-octyl]-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [543] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and the product of Step A as the appropriate alkylating agent, 13 mg of the desired product were obtained.
• Step A 3-[5-[1-(12-bromododecanoyl)-4-piperidyl]-1-oxo-isoindolin-2-yl]piperidine-2,6- dione [544] Using General procedure for the acylation of piperidinyl-isoindolinone starting from 3- [1-oxo-5-(4-piperidyl)isoindolin-2-yl]piperidine-2,6-dione, hydrochloride (1:1) (0.42 mmol) and 12-bromododecanoic acid as the appropriate acid, 199 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[12-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindolin-5-yl]-1- piperidyl]-12-oxo-dodecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [545] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and the product of Step A as the appropriate alkylating agent, 17 mg of the desired product were obtained.
• Step A 3-[5-[1-(10-bromodecanoyl)-4-piperidyl]-1-oxo-isoindolin-2-yl]piperidine-2,6- dione [546] Using General procedure for the acylation of piperidinyl-isoindolinone starting from 3- [1-oxo-5-(4-piperidyl)isoindolin-2-yl]piperidine-2,6-dione, hydrochloride (1:1) (0.42 mmol) and 10-bromodecanoic acid as the appropriate acid, 71 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[10-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindolin-5-yl]-1- piperidyl]-10-oxo-decyl]-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [547] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and the product of Step A as the appropriate alkylating agent, 14 mg of the desired product were obtained.
• Step A 3-[5-[1-(9-bromononanoyl)-4-piperidyl]-1-oxo-isoindolin-2-yl]piperidine-2,6- dione
• hydrochloride (1:1) (0.42 mmol)
• 9-bromononanoic acid 9-bromononanoic acid
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[9-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindolin-5-yl]-1- piperidyl]-9-oxo-nonyl]-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [549] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and the product of Step A as the appropriate alkylating agent, 18 mg of the desired product were obtained.
• Step A (2S,4R)-N-[[2-(9-bromononoxy)-4-(4-methylthiazol-5-yl)phenyl]methyl]-1-[(2S)- 2-[(1-fluorocyclopropanecarbonyl)amino]-3,3-dimethyl-butanoyl]-4-hydroxy- pyrrolidine-2-carboxamide [550] Using General procedure for the alkylation of VHL ligand on hydroxy group starting from (2S,4R)-1-[(2S)-2-[(1-fluorocyclopropanecarbonyl)amino]-3,3-dimethyl-butanoyl]-4-hydroxy- N-[[2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (0.38 mmol) and 1,9-dibromononane as the appropriate reactant, 147 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[9-[2-[[[(2S,4R)-1-[(2S)-2-[(1- fluorocyclopropanecarbonyl)amino]-3,3-dimethyl-butanoyl]-4-hydroxy-pyrrolidine-2- carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]nonyl-methyl-amino]ethoxy]- 5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [551] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and the product of Step A
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[10-[2-[[[[(2S,4R)-1-[(2S)-2-[(1- fluorocyclopropanecarbonyl)amino]-3,3-dimethyl-butanoyl]-4-hydroxy-pyrrolidine-2- carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]decyl-methyl-amino]ethoxy]- 5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [553] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and the product of Step
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[16-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindolin-5-yl]-1- piperidyl]-16-oxo-hexadecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]- 5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [555] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and the product of Step A as the appropriate alkylating agent, 8 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[11-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindolin-5-yl]-1- piperidyl]-11-oxo-undecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [557] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[11-[2-[[[[(2S,4R)-1-[(2S)-2-[(1- fluorocyclopropanecarbonyl)amino]-3,3-dimethyl-butanoyl]-4-hydroxy-pyrrolidine-2- carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]undecyl-methyl- amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2- carboxylic acid [559] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and the product of Step A as
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[16-[2-[[[[(2S,4R)-1-[(2S)-2-[(1- fluorocyclopropanecarbonyl)amino]-3,3-dimethyl-butanoyl]-4-hydroxy-pyrrolidine-2- carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]hexadecyl-methyl- amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyr
• Step B [13-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindolin-5-yl]-1-piperidyl]-13-oxo- tridecyl] 4-methylbenzenesulfonate [563] Using General procedure for the tosylation of the hydroxyalkyl VHL ligand-derivatives starting from product of Step A (0.32 mmol), 56 mg of the desired product were obtained.
• Step C 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[13-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindolin-5-yl]-1- piperidyl]-13-oxo-tridecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [564] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (25 mg) and the product of Step B as the appropriate alkylating agent, 4 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[12-[(2R)-2-[(1-fluorocyclopropanecarbonyl)amino]-3- [(2S,4R)-4-hydroxy-2-[[4-(4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidin-1- yl]-1,1-dimethyl-3-oxo-propyl]sulfanyldodecyl-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [566] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[14-[(2R)-2-[(1-fluorocyclopropanecarbonyl)amino]-3- [(2S,4R)-4-hydroxy-2-[[4-(4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidin-1- yl]-1,1-dimethyl-3-oxo-propyl]sulfanyltetradecyl-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [568] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[8-[4-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]carbamoyl]-1-piperidyl]-8-oxo-octyl]-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [570] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[11-[4-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]carbamoyl]-1-piperidyl]-11-oxo-undecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[15-[2-[[[[(2S,4R)-1-[(2S)-2-[(1- fluorocyclopropanecarbonyl)amino]-3,3-dimethyl-butanoyl]-4-hydroxy-pyrrolidine-2- carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]pentadecyl-methyl- amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyr
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[10-[4-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]carbamoyl]-1-piperidyl]-10-oxo-decyl]-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[13-[2-[[[[(2S,4R)-1-[(2S)-2-[(1- fluorocyclopropanecarbonyl)amino]-3,3-dimethyl-butanoyl]-4-hydroxy-pyrrolidine-2- carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]tridecyl-methyl- amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2- carboxy
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[12-[4-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]carbamoyl]-1-piperidyl]-12-oxo-dodecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1- adamant
• Step B 2-[2-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]acetic acid [582] A mixture of 1.60 g of the product of Step A, DCM (6.0 mL) and TFA (2.4 mL, 9 eq) was stirred at RT for 18 h.1.25 g of the desired product was obtained after removal the volatiles.
• Step C 2-[2-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo- ethoxy]ethoxy]ethoxy]ethoxy]ethyl 4-methylbenzenesulfonate [583] Using General procedure for the acylation of VHL ligands starting from 300 mg of (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl] pyrrolidine-2-carboxamide, hydrogen chloride (1:1) (0.62 mmol) and the product
• Step D 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[2-[2-[2-[2-[2-[2-[[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-2-oxo-ethoxy]ethoxy]ethoxy]ethoxy]ethyl-methyl-amino]ethoxy]-5,7- dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [584] Using Degrader Synthesis by Alky
• Step A (2S,4R)-1-[(2S)-2-[3-[2-[2-(2-bromoethoxy)ethoxy]ethoxy]propanoylamino]-3,3- dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]pyrrolidine-2-carboxamide [585] Using General procedure for the acylation of VHL ligands starting from 300 mg (0.62 mmol) of (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl] pyrrolidine-2-carboxamide, hydrogen chloride (1:1) and 3-[2- [2-(2-bromoethoxy)ethoxy]ethoxy]propanoic acid
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[2-[2-[2-[2-[3-[[(1S)-1-[(2R,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-3-oxo-propoxy]ethoxy]ethoxy]ethyl-methyl-amino]ethoxy]-5,7-dimethyl- 1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [586] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure
• Step B 2-[2-[2-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]acetic acid [588] A mixture of 1.30 g of the product of Step A, DCM (6.0 mL) and TFA (1.8 mL, 9 eq) was stirred at RT for 18 h.1.00 g of the desired product was obtained after removal the volatiles.
• Step C 2-[2-[2-[2-[3-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-3-oxo- propoxy]ethoxy]ethoxy]ethyl 4-methylbenzenesulfonate [589] Using General procedure for the acylation of VHL ligands from 300 mg (0.62 mmol) of (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl] ethyl] pyrrolidine-2-carboxamide, hydrogen chloride (1:1) and the product of
• Step D 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[2-[2-[2-[2-[2-[2-[3-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethyl-methyl-amino]ethoxy]-5,7- dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [590] Using Degrader Synthesis by Al
• Step B 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[11-[[2-(2,6- dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]undecanoyl-methyl- amino]butyl]amino]-3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [592] Using Degrader Synthesis by Amide Coupling and Hydrolysis General Procedure starting from the product of Step A as the appropriate acid and 40 mg (0.04 mmol) of Preparation 19 as the appropriate amine, 20 mg of the desired product were obtained.
• Step B 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[9-[[2-(2,6-dioxo- 3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]nonanoyl-methyl-amino]butyl]amino]-3-[1- [[3-[2-(dimethylamino)ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carboxylic acid [594] Using Degrader Synthesis by Amide Coupling General Procedure and Hydrolysis General Procedure starting from the product of Step A as the appropriate acid and 40 mg (0.04 mmol) of Preparation 19 as the appropriate amine, 31 mg of the desired product were obtained.
• Step B 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[10-[[2-(2,6- dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]decanoyl-methyl- amino]butyl]amino]-3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [596] Using Degrader Synthesis by Amide Coupling and Hydrolysis General Procedure starting from the product of Step A as the appropriate acid and 40 mg (0.04 mmol) of Preparation 19 as the appropriate amine, 49 mg of the desired product were obtained.
• Step B 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[12-[[2-(2,6- dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]amino]dodecanoyl-methyl- amino]butyl]amino]-3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [598] Using Degrader Synthesis by Amide Coupling and Hydrolysis General Procedure starting from the product of Step A as the appropriate acid and 40 mg (0.04 mmol) of Preparation 19 as the appropriate amine, 20 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[10-[(2R)-2-[(1-fluorocyclopropanecarbonyl)amino]-3- [(2S,4R)-4-hydroxy-2-[[4-(4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidin-1- yl]-1,1-dimethyl-3-oxo-propyl]sulfanyldecyl-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [600] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Prepar
• Step B 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[10-[2-(2,6-dioxo- 3-piperidyl)-1,3-dioxo-isoindolin-4-yl]oxydecanoyl-methyl-amino]butyl]amino]-3-[1-[[3- [2-(dimethylamino)ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carboxylic acid [602] Using Degrader Synthesis by Amide Coupling and Hydrolysis General Procedure starting from the product of Step A as the appropriate acid and 40 mg (0.04 mmol) of Preparation 19 as the appropriate amine, 44 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[12-[3-[2-[(2S)-1-[(2S)-2-cyclohexyl-2-[[(2S)-2- (methylamino)propanoyl]amino]acetyl]pyrrolidin-2-yl]thiazole-4- carbonyl]phenoxy]dodecyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [604] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step A as the appropriate bromine and 50 mg (0.06 mmol) of Preparation 4 as the appropriate
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[8-[3-[2-[(2S)-1-[(2S)-2-cyclohexyl-2-[[(2S)-2- (methylamino)propanoyl]amino]acetyl]pyrrolidin-2-yl]thiazole-4- carbonyl]phenoxy]octyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [606] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step A as the appropriate bromine and 40 mg (0.05 mmol) of Preparation 4 as the appropriate amine
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[10-[3-[2-[(2S)-1-[(2S)-2-cyclohexyl-2-[[(2S)-2- (methylamino)propanoyl]amino]acetyl]pyrrolidin-2-yl]thiazole-4- carbonyl]phenoxy]decyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [608] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step A as the appropriate bromine and 30 mg (0.04 mmol) of Preparation 4 as the appropriate amine
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[4-[3-[2-[(2S)-1-[(2S)-2-cyclohexyl-2-[[(2S)-2- (methylamino)propanoyl]amino]acetyl]pyrrolidin-2-yl]thiazole-4- carbonyl]phenoxy]butyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [610] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step A as the appropriate bromine and 40 mg (0.05 mmol) of Preparation 4 as the appropriate amine, 43 mg
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[14-[3-[2-[(2S)-1-[(2S)-2-cyclohexyl-2-[[(2S)-2- (methylamino)propanoyl]amino]acetyl]pyrrolidin-2-yl]thiazole-4- carbonyl]phenoxy]tetradecyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]- 5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [612] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step A as the appropriate bromine and 40 mg (0.05 mmol) of Preparation 4 as the appropriate amine
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[6-[3-[2-[(2S)-1-[(2S)-2-cyclohexyl-2-[[(2S)-2- (methylamino)propanoyl]amino]acetyl]pyrrolidin-2-yl]thiazole-4- carbonyl]phenoxy]hexyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [616] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step A as the appropriate bromine and 40 mg (0.05 mmol) of Preparation 4 as the appropriate amine,
• Step B 3-[2-(p-tolylsulfonyloxy)ethoxy]propanoic acid [618]
• the product of Step A (1 g, 2.9 mmol) in DCM (14.5 mL) was treated with TFA (7.5 eq) at 0 °C and the mixture was stirred at room temperature until complete conversion was observed. The product was concentrated and used without further purification (802 mg, 95%).
• Step C 2-[3-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-3-oxo- propoxy]ethyl 4-methylbenzenesulfonate [619] Using General procedure for the acylation and deprotection of VHL ligands without the hydrolysis step, starting from 300 mg of (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4- hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] pyrrolidine-2-carboxamide, hydrogen chloride (1:1) (0.62 mmol) and the product of Step B as the appropriate acid,
• Step D 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[2-[3-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-3-oxo-propoxy]ethyl-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [620] Using General procedure for production of VHL ligand-based degraders via alkylation starting from the product of Step C as the
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[9-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5- yl]oxynonyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl- pyrazol-4-yl]pyridine-2-carboxylic acid [622] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step A as the appropriate bromide and 50 mg (0.06 mmol) of Preparation 4 as the appropriate amine, 44 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[11-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5- yl]oxyundecyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl- pyrazol-4-yl]pyridine-2-carboxylic acid [624] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step A as the appropriate bromide and 50 mg (0.06 mmol) of Preparation 4 as the appropriate amine, 49 mg of the desired product were obtained.
• Step A 5-(7-bromoheptoxy)-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[7-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5- yl]oxyheptyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl- pyrazol-4-yl]pyridine-2-carboxylic acid [626] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step A as the appropriate bromide and 50 mg (0.06 mmol) of Preparation 4 as the appropriate amine, 45 mg of the desired product were obtained.
• Step A 5-(5-bromopentoxy)-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[5-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5- yl]oxypentyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl- pyrazol-4-yl]pyridine-2-carboxylic acid [628] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step A as the appropriate bromide and 75 mg (0.09 mmol) of Preparation 4 as the appropriate amine, 42 mg of the desired product were obtained.
• Step A tert-butyl 3-(6-hydroxyhexoxy)propanoate
• hexane-1,6-diol 8.46 mmol
• benzyl(trimethyl)ammonium hydroxide (1:1) (0.77 mL, 1.69 mmol)
• tert-butyl prop-2-enoate 1.5 mL, 10.15 mmol
• the reaction was quenched with brine and extracted with DCM to give 100 mg of the desired product.
• Step B tert-butyl 3-[6-(p-tolylsulfonyloxy)hexoxy]propanoate [630] Using General procedure for the tosylation of the hydroxyalkyl VHL ligand-derivatives starting from 285 mg of the product of Step A, 333 mg of the desired product were obtained.
• Step C 3-[6-(p-tolylsulfonyloxy)hexoxy]propanoic acid [631]
• the product of Step B (333 mg) in DCM (4 mL) was treated with TFA (7.5 eq) at 0 °C and the mixture was stirred at room temperature until complete conversion was observed. The product was concentrated and used without further purification (280 mg, 97%).
• Step D 6-[3-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-3-oxo- propoxy]hexyl 4-methylbenzenesulfonate [632] Using General procedure for the acylation and deprotection of VHL ligands without the hydrolysis step starting from 250 mg of (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4- hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] pyrrolidine-2-carboxamide, hydrogen chloride (1:1) (0.52 mmol) and the product of Step C as the appropriate acid, 130
• Step E 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[6-[3-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-3-oxo-propoxy]hexyl-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-
• Step B tert-butyl 2-[9-(p-tolylsulfonyloxy)nonoxy]acetate
• the product was purified by column chromatography using heptane and ethyl acetate as eluents to give 167 mg of the desired product.
• Step C 2-[9-(p-tolylsulfonyloxy)nonoxy]acetic acid [636]
• the product of Step B (167 mg) in DCM (2 mL) was treated with TFA (7.5 eq) at 0 °C and the mixture was stirred at room temperature until complete conversion was observed. The product was concentrated and used without further purification (142 mg, 97%).
• Step D 9-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo- ethoxy]nonyl 4-methylbenzenesulfonate [637] Using General procedure for the acylation and deprotection of VHL ligands without the hydrolysis step starting from 125 mg of (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4- hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] pyrrolidine-2-carboxamide, hydrogen chloride (1:1) (0.26 mmol) and the product of Step C as the appropriate
• Step E 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[9-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-2-oxo-ethoxy]nonyl-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [638] Using General procedure for production of VHL ligand-based degraders via alkylation starting from the product of Step D
• Step B tert-butyl 2-[11-(p-tolylsulfonyloxy)undecoxy]acetate
• diacetoxyrhodium 2.65 mg, 0.015 eq
• tert-butyl 2-diazoacetate 1.14 g, 1.2 mmol
• Step C 2-[11-(p-tolylsulfonyloxy)undecoxy]acetic acid [641]
• the product of Step B (261 mg) in DCM (2 mL) was treated with TFA (7.5 eq) at 0 °C and the mixture was stirred at room temperature until complete conversion was observed. The product was concentrated and used without further purification (225 mg).
• Step D 11-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo- ethoxy]undecyl 4-methylbenzenesulfonate [642] Using General procedure for the acylation and deprotection of VHL ligands without the hydrolysis step starting from 200 mg of (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4- hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] pyrrolidine-2-carboxamide, hydrogen chloride (1:1) (0.42 mmol) and the product of Step C as the appropriate
• Step E 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[11-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-2-oxo-ethoxy]undecyl-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [643] Using General procedure for production of VHL ligand-based degraders via alkylation starting from the product of Step
• Step B tert-butyl 2-[13-(p-tolylsulfonyloxy)tridecoxy]acetate [645]
• diacetoxyrhodium 2.0 mg, 0.015 mmol
• tert-butyl 2-diazoacetate 128 mg, 0.90 mmol
• the product was purified by column chromatography using heptane and ethyl acetate as eluents to give 145 mg of the desired product.
• Step C 2-[13-(p-tolylsulfonyloxy)tridecoxy]acetic acid [646]
• the product of Step B (145 mg) in DCM (2 mL) was treated with TFA (7.5 eq) at 0 °C and the mixture was stirred at room temperature until complete conversion was observed. The product was concentrated and used without further purification (126 mg).
• Step D 13-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo- ethoxy]tridecyl 4-methylbenzenesulfonate [647] Using General procedure for the acylation and deprotection of VHL ligands without the hydrolysis step starting from 125 mg of (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4- hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] pyrrolidine-2-carboxamide, hydrogen chloride (1:1) (0.26 mmol) and the product of Step C as the
• Step E 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[13-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-2-oxo-ethoxy]tridecyl-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [648] Using General procedure for production of VHL ligand-based degraders via alkylation starting from the product of Step
• Step B 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[[12-[[(1S)-1- [(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-12-oxo- dodecanoyl]-methyl-amino]butyl]amino]-3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7- dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [650] Using Degrader Synthesis by Amide Coupling and Hydrolysis General Procedure starting from the product of Step A as the appropriate acid and 40 mg (
• Step B 2-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]acetic acid [652]
• TFA 0.68 mL, 7.5 eq
• the reaction mixture was stirred for 18 h.430 mg of the product was isolated after removing the volatiles under reduced pressure.
• Step C 2-[2-[2-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-2-oxo- ethoxy]ethoxy]ethoxy]ethyl 4-methylbenzenesulfonate [653] Using General procedure for the acylation and deprotection of VHL ligands without the hydrolysis step starting from 250 mg of (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4- hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] pyrrolidine-2-carboxamide, hydrogen chloride (1:1) (0.52 mmol
• Step D 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[2-[2-[2-[2-[2-[[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-2-oxo-ethoxy]ethoxy]ethoxy]ethyl-methyl-amino]ethoxy]-5,7-dimethyl- 1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [654] Using General procedure for production of VHL ligand-based de
• Step A 11-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-11-oxo- undecanoic acid [655] Using General procedure for the acylation and deprotection of VHL ligands starting from (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl] pyrrolidine-2-carboxamide, hydrogen chloride (1:1) (0.62 mmol) and 11-tert- butoxy-11-oxo-undecanoic acid as the appropriate acid, 152 mg of the desired product were obtained
• Step B 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[[11-[[(1S)-1- [(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-11-oxo- undecanoyl]-methyl-amino]butyl]amino]-3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7- dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [
• Step A 5-(13-bromotridecoxy)-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[13-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5- yl]oxytridecyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl- pyrazol-4-yl]pyridine-2-carboxylic acid [658] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step A as the appropriate bromide and 35 mg (0.04 mmol) of Preparation 4 as the appropriate amine, 11 mg of the desired product were obtained.
• Step A 14-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-14-oxo- tetradecanoic acid [659] Using General procedure for the acylation and deprotection of VHL ligands starting from 300 mg (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl] pyrrolidine-2-carboxamide, hydrogen chloride (1:1) (0.62 mmol) and 14-tert-butoxy-14-oxo-tetradecanoic acid as the appropriate acid, 455 mg
• Step B 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[[14-[[(1S)-1- [(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-14-oxo- tetradecanoyl]-methyl-amino]butyl]amino]-3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7- dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-car
• Step A (2S,4R)-1-[(2R)-3-(11-bromoundecylsulfanyl)-2-[(1- fluorocyclopropanecarbonyl)amino]-3-methyl-butanoyl]-4-hydroxy-N-[[4-(4- methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide [661] Using General procedure for the alkylation of VHL ligand on thiol group starting from 75 mg of (2S,4R)-1-[(2R)-2-[(1-fluorocyclopropanecarbonyl)amino]-3-methyl-3-sulfanyl- butanoyl]-4-hydroxy-N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (0.14 mmol) and 1,11-dibromoundecane as the appropriate dibromide, 97.4 mg
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[11-[(2R)-2-[(1-fluorocyclopropanecarbonyl)amino]-3- [(2S,4R)-4-hydroxy-2-[[4-(4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidin-1- yl]-1,1-dimethyl-3-oxo-propyl]sulfanylundecyl-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [662] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[13-[(2R)-2-[(1-fluorocyclopropanecarbonyl)amino]-3- [(2S,4R)-4-hydroxy-2-[[4-(4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidin-1- yl]-1,1-dimethyl-3-oxo-propyl]sulfanyltridecyl-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [664] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[15-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5- yl]oxypentadecyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl- pyrazol-4-yl]pyridine-2-carboxylic acid [666] Using Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Step A as the appropriate bromide and 40 mg (0.05 mmol) of Preparation 4 as the appropriate amine, 15 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3,5-dimethyl-7-(2-pyrrolidin-1-ylethoxy)-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]-N-[7-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-7-oxo- heptyl]pyridine-2-carboxamide [668] Using Degrader Synthesis by Amide Coupling General Procedure starting from the product of Step A as the appropriate amine and 30 mg (0.
• Step B 13-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-13-oxo- tridecanoic acid [670]
• the mixture of 89 mg (0.12 mmol) of the product of Step A in 0.6 mL of THF and 0.1 mL of water was treated with 10 eq of lithium hydroxide at 50 °C for 5 h.
• Step C 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[[13-[[(1S)-1- [(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-13-oxo- tridecanoyl]-methyl-amino]butyl]amino]-3-[1-[[3-[2-(dimethylamino)ethoxy]-5,7- dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [671] Using Degrader Synthesis by Amide Coupling and Hydrolysis General Procedure starting from the product of Step B as the appropriate acid and 50 mg
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3,5-dimethyl-7-(2-pyrrolidin-1-ylethoxy)-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]-N-[5-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-5-oxo- pentyl]pyridine-2-carboxamide [
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3,5-dimethyl-7-(2-pyrrolidin-1-ylethoxy)-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]-N-[3-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-3-oxo- propyl]pyridine-2-carboxamide [675] Using Degrader Synthesis by Amide Coupling General Procedure starting from the product of Step A as the appropriate amine and 43 mg (0.04
• Step A (2S,4R)-1-[(2S)-2-(9-aminononanoylamino)-3,3-dimethyl-butanoyl]-4-hydroxy- N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3,5-dimethyl-7-(2-pyrrolidin-1-ylethoxy)-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]-N-[9-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-9-oxo- nonyl]pyridine-2-carboxamide [677] Using Degrader Synthesis by Amide Coupling General Procedure starting from the product of Step A as the appropriate amine and 45 mg (0.05 m
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3,5-dimethyl-7-(2-pyrrolidin-1-ylethoxy)-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]-N-[11-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-11-oxo- undecyl]pyridine-2-carboxamide [679] Using Degrader Synthesis by Amide Coupling General Procedure starting from the product of Step A as the appropriate amine and 43 mg (0.04
• Step B 13-[(2R)-2-[(1-fluorocyclopropanecarbonyl)amino]-3-[(2S,4R)-4-hydroxy-2-[[4- (4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidin-1-yl]-1,1-dimethyl-3-oxo- propyl]sulfanyltridecanoic acid [683] The mixture of 84 mg (0.11 mmol) of the product of Step A in 0.6 mL of THF and 0.1 mL of water was treated with 10 eq of lithium hydroxide at 50 °C for 5 h.
• Step C 6-[[6-(1,3-benzothiazol-2-ylamino)-5-methyl-pyridazin-3-yl]-[4-[13-[(2R)-2-[(1- fluorocyclopropanecarbonyl)amino]-3-[(2S,4R)-4-hydroxy-2-[[4-(4-methylthiazol-5- yl)phenyl]methylcarbamoyl]pyrrolidin-1-yl]-1,1-dimethyl-3-oxo- propyl]sulfanyltridecanoyl-methyl-amino]butyl]amino]-3-[1-[[3-[2- (dimethylamino)ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carboxylic acid [684] Using Degrader Synthesis by Amide Coupling and Hydrolysis General Procedure starting from the
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3,5-dimethyl-7-(2-pyrrolidin-1-ylethoxy)-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]-N-[8-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-8-oxo- octyl]pyridine-2-carboxamide [687] Using Degrader Synthesis by Amide Coupling General Procedure starting from the product of Step A as the appropriate amine and 48 mg (0
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3,5-dimethyl-7-(2-pyrrolidin-1-ylethoxy)-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]-N-[10-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-10-oxo- decyl]pyridine-2-carboxamide [689] Using Degrader Synthesis by Amide Coupling General Procedure starting from the product of Step A as the appropriate amine and 46 mg (0.06
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3,5-dimethyl-7-(2-pyrrolidin-1-ylethoxy)-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]-N-[12-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-12-oxo- dodecyl]pyridine-2-carboxamide [691] Using Degrader Synthesis by Amide Coupling General Procedure starting from the product of Step A as the appropriate amine and 44 mg (0.05
• Step B 2-(2,6-dioxo-3-piperidyl)-4-[2-[2-[2-[2-(2- iodoethoxy)ethoxy]ethoxy]ethylamino]isoindoline-1,3-dione [693] Using the General procedure for the iodination of hydroxyalkyl derivative of thalidomide starting from the product of Step A (35 mg), 18 mg of the desired product were obtained.
• Step B [10-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-10-oxo- decyl] 4-methylbenzenesulfonate [696] Using the General procedure for the tosylation of the hydroxyalkyl VHL ligand- derivatives starting from the product of Step A (100 mg), 90 mg of the desired product were obtained.
• Step C 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[10-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-10-oxo-decyl]-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]- 5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [697] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (22 mg) and the
• Example 135 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[11-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-11-oxo-undecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid Step A: (2S,4R)-4-hydroxy-1-[(2S)-2-(11-hydroxyundecanoylamino)
• Step B [11-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-ylphenyl]ethyl] carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-11-oxo-undecyl] 4- methyl benzenesulfonate [700] Using the General procedure for the tosylation of the hydroxyalkyl VHL ligand- derivatives starting from the product of Step A (100 mg), 95 mg of the desired product were obtained.
• Step C 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[11-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-11-oxo-undecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [701] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and
• Step A (2S,4R)-4-hydroxy-1-[(2S)-2-(13-hydroxytridecanoylamino)-3,3-dimethyl- butanoyl]-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide
• hydrogen chloride (1:1) (0.42 mmol
• 13-hydroxytridecanoic acid as the appropriate acid
• Step B [13-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-13-oxo- tridecyl] 4-methylbenzenesulfonate [703] Using the General procedure for the tosylation of the hydroxyalkyl VHL ligand- derivatives starting from the product of Step A (100 mg), 88 mg of the desired product were obtained.
• Step C 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[[13-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-13-oxo-tridecyl]-methyl-amino]ethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [704] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[6-[2-[[[[(2S,4R)-1-[(2S)-2-[(1- fluorocyclopropanecarbonyl)amino]-3,3-dimethyl-butanoyl]-4-hydroxy-pyrrolidine-2- carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]hexyl-methyl-amino]ethoxy]- 5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [706] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and the product of Step
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[8-[2-[[[(2S,4R)-1-[(2S)-2-[(1- fluorocyclopropanecarbonyl)amino]-3,3-dimethyl-butanoyl]-4-hydroxy-pyrrolidine-2- carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]octyl-methyl-amino]ethoxy]- 5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid [708] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (25 mg) and the product of
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[12-[2-[[[[(2S,4R)-1-[(2S)-2-[(1- fluorocyclopropanecarbonyl)amino]-3,3-dimethyl-butanoyl]-4-hydroxy-pyrrolidine-2- carbonyl]amino]methyl]-5-(4-methylthiazol-5-yl)phenoxy]dodecyl-methyl- amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2- carboxylic acid [710] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (13 mg) and the product of Step
• Step A 5-(12-bromododecoxy)-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione [711] Using the General procedure for the alkylation of the 5-hydroxy thalidomide starting from 2-(2,6-dioxo-3-piperidyl)-5-hydroxy-isoindoline-1,3-dione (0.44 mmol) and 1,12- dibromododecane as the appropriate bromoalkane, 96 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[12-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5- yl]oxydodecyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl- pyrazol-4-yl]pyridine-2-carboxylic acid [712] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and the product of Step A as the appropriate alkylating agent, 27 mg of the desired product were obtained.
• Step A 4-(6-bromohexoxy)-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione [713] Using the General procedure for the alkylation of the 5-hydroxy thalidomide starting from 2-(2,6-dioxo-3-piperidyl)-4-hydroxy-isoindoline-1,3-dione (0.44 mmol) and 1,6- dibromohexane as the appropriate bromoalkane, 52 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[6-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4- yl]oxyhexyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol- 4-yl]pyridine-2-carboxylic acid [714] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (35 mg) and the product of Step A as the appropriate alkylating agent, 25 mg of the desired product were obtained.
• Step A 5-(14-bromotetradecoxy)-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione [715] Using the General procedure for the alkylation of the 5-hydroxy thalidomide starting from 2-(2,6-dioxo-3-piperidyl)-5-hydroxy-isoindoline-1,3-dione (0.44 mmol) and 1,14- dibromotetradecane as the appropriate bromoalkane, 70 mg of the desired product were obtained.
• Step B 6-[3-(1,3-benzothiazol-2-ylamino)-4-methyl-6,7-dihydro-5H-pyrido[2,3- c]pyridazin-8-yl]-3-[1-[[3-[2-[14-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5- yl]oxytetradecyl-methyl-amino]ethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl- pyrazol-4-yl]pyridine-2-carboxylic acid [716] Using the Degrader Synthesis by Alkylation and Hydrolysis General Procedure starting from the product of Preparation 4 (30 mg) and the product of Step A as the appropriate alkylating agent, 13 mg of the desired product were obtained.
• Step A 4-(8-bromooctoxy)-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione [717] Using the General procedure for the alkylation of the 5-hydroxy thalidomide starting from 2-(2,6-dioxo-3-piperidyl)-4-hydroxy-isoindoline-1,3-dione (0.44 mmol) and 1,8- dibromooctane as the appropriate bromoalkane, 140 mg of the desired product were obtained.

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