Patent Application
20240307547
The present invention relates to novel compounds which bind to the protein cereblon and modulate the substrate specificity of CUL4-DDB1-RBX1-CRBN ubiquitin ligase complex (CRL4CRBN) Cereblon is a substrate recognition component of CRL4CRBN. Chemical modulation of cereblon may induce association of novel substrate proteins, followed by their ubiquitination and degradation.
Cereblon (CRBN) is a protein which associates with DDB1 (damaged DNA binding protein 1), CUL4 (Cullin-4), and RBX1 (RING-Box Protein 1). Collectively, the proteins form a ubiquitin ligase complex, which belongs to Cullin RING Ligase (CRL) protein family and is referred to as CRL4CRBN Cereblon became of particular interest to the scientific community after it was confirmed to be a direct protein target of thalidomide, which mediates the biological activity of cereblon. Thalidomide, a drug approved for treatment of multiple myeloma in the late 1990s, binds to cereblon and modulates the substrate specificity of the CRL4CRBN ubiquitin ligase complex. This mechanism underlies the pleiotropic effect of thalidomide on both immune cells and cancer cells (see Lu G et al.: The Myeloma Drug Lenalidomide Promotes the Cereblon-Dependent Destruction of Ikaros Proteins. Science. 2014 Jan. 17; 343(6168): 305-9).
Thalidomide's success in cancer therapy stimulated efforts towards development of analogues with higher potency and fewer side effects. As a results, various drug candidates were produced: lenalidomide, pomalidomide, CC-220, CC-122, CC-885, and TD-106. These compounds are collectively called Cereblon Modulating Agents (CMAs). For discussions of these compounds, see—for example—U.S. Pat. No. 5,635,517(B2), WO2008039489 (A2), WO2017197055 (A1), WO2018237026 (A1), WO2017197051 (A1), U.S. Pat. No. 8,518,972 (B2), EP 2057143 (B1), WO2019014100 (A1), WO2004103274 (A2), and Kim S A et al.: A novel cereblon modulator for targeted protein degradation. Eur J Med Chem. 2019 Mar. 15; 166: 65-74.
The clinical applicability of CMAs in numerous hematologic malignancies, such as multiple myeloma, myelodysplastic syndromes lymphomas and leukemia, has been demonstrated (see Le Roy A et al.: Immunomodulatory Drugs Exert Anti-Leukemia Effects in Acute Myeloid Leukemia by Direct and Immunostimulatory Activities. Front Immunol. 2018; 9: 977).
The antitumor activity of cereblon modulators is mediated by:
It has been demonstrated that chemically-modified thalidomide-based derivatives can significantly modify the substrate specificity of CRL4CRBN ubiquitin ligase. Thus, it is desired to progress development of cereblon modulating agents in order to achieve desired substrate specificity in the CMA-bound CRL4CRBN ubiquitin ligase complex (see Sievers Q L et al.: Defining the human C2H2 zinc finger degrome targeted by thalidomide analogues through CRBN. Science. 2018 Nov. 2; 362(6414)) to reach a desired safety profile as well as to achieve stable drugs without loss of their potency due to chemical degradation. There is thus a continuing need to provide novel cereblon-binding compounds which have pharmaceutically relevant properties while ensuring their high drug stability with improved resistance to hydrolytic degradation.
In accordance with a first aspect of the invention, there is provided a compound of Formula (I):
or a pharmaceutically acceptable salt, ester, optically active isomer, racemate, solvate, amino acid conjugate, or prodrug thereof,
In certain embodiments, the compound of Formula (I) has the structure:
In other embodiments, the compound of Formula (I) has the structure:
In some embodiments, each R′ is independently hydrogen, halogen, —NH2, —NO2, —C(O)NHCHR″2, —CHR″NHC(O)NHR″, —CHR″NHC(O)C(halogen)2R″ or —NHS(O)2R″.
In some embodiments, each R″ is independently hydrogen, alkyl, cycloalkyl, or aryl. In some such embodiments, the aryl is substituted with one or more groups selected from halogen, alkyl and O-haloalkyl. In some embodiments, the halogen is Cl, the alkyl is methyl and the O-haloalkyl is O—CF3.
In some embodiments, one of W1, W2, W3 and W4 is N, and the remaining three of W1, W2, W3 and W4 are each CR′. In some such embodiments, W1 is N, and W2, W3 and W4 are CR′. In other embodiments, W2 is N, and W1, W3 and W4 are CR′. In yet other embodiments, W3 is N, and W1, W2 and W4 are CR′. In other embodiments, W4 is N, and W1, W2 and W3 are CR′.
In some embodiments, W1, W2, W3 and W4 are each CR′.
In some embodiments, W1, W2, W3 and W4 are each CH.
In other embodiments, three of W1, W2, W3 and W4 are CH, and one of W1, W2, W3 and W4 is C-halogen, C-alkyl, C-alkenyl, C-alkynyl, C-aryl, C-heteroaryl, C-benzyl, C-haloalkyl, C-haloalkenyl, C—NH2, C—NHR″, C—NR″2, C—NR″C(O)R″, C—NR″C(O)OR″, C—NO2, C—CN, C—C(O)R″, C—C(O)OR″, C—C(O)NH2, C—C(O)NHR″, C—C(O)NR″2, C—C(O)NHCHR″2, C—CHR″NHC(O)NHR″, C—CHR″NHC(O)C(halogen)2R″, C—OR″, C—OC(O)R″, C—OC(O)OR″, C—OC(O)NH2, C—OC(O)NHR″, C—OC(O)NR″2, C—SR″, C—S(O)2R″, C—S(O)2OR″, C—S(O)2NH2, C—S(O)2NHR″, C—S(O)2NR″2, or C—NHS(O)2R″. In some such embodiments, one of W1, W2, W3 and W4 is C-halogen, C—NH2, C—NO2, C—NHR″, C—NR″2, C—C(O)NHCHR″2, C—CHR″NHC(O)NHR″, C—CHR″NHC(O)C(halogen)2R″ or C—NHS(O)2R″. In some such embodiments, one of W1, W2, W3 and W4 is C-halogen, C—NH2, C—NO2, C—C(O)NHCHR″2, C—CHR″NHC(O)NHR″, C—CHR″NHC(O)C(halogen)2R″ or C—NHS(O)2R″. In some such embodiments, one of W1, W2, W3 and W4 is C-halogen, C—NH2, C—NO2, C—C(O)NHCHR″2, C—CH2NHC(O)NHR″, C—CH2NHC(O)CF2R″ or C—NHS(O)2R″.
In some embodiments W2, W3 and W4 are each CH. In some such embodiments, W1 is C-halogen, C—NH2, C—NO2 or C—NHS(O)2R″. In some such embodiments, W1 is C—NH2 or C—NHS(O)2R″.
In other embodiments W1, W2 and W3 are each CH. In some such embodiments, W4 is C-halogen, C—NH2, C—NO2 or C—NHS(O)2R″. In some such embodiments, W4 is C—NH2.
In other embodiments W1, W2 and W4 are each CH. In some such embodiments, W2 is C—NH2, C—NO2 or C—NHS(O)2R″. In some such embodiments, W2 is C—NH2 or C—NHS(O)2R″.
In other embodiments W1, W3 and W4 are each CH. In some such embodiments, W3 is C—NH2, C—NO2, C—C(O)NHCHR″2, C—CH2NHC(O)NHR″, C—CH2NHC(O)CF2R″ or C—NHS(O)2R″. In some such embodiments, W3 is C—NH2, C—C(O)NHCHR″2, C—CH2NHC(O)NHR″, C—CH2NHC(O)CF2R″ or C—NHS(O)2R″. In some such embodiments, W3 is C—NH2, C—CH2NHC(O)NHR″, C—CH2NHC(O)CF2R″ or C—NHS(O)2R″.
In some embodiments, Q1 is N and Q2 is CR.
In other embodiments, Q1 is N and Q2 is N In other embodiments, Q1 is CR and Q2 is N. In some such embodiments, Q1 is C—H or C-alkyl. In some such embodiments, Q1 is C—H. In other embodiments, Q1 is C-methyl.
In other embodiments, two of W1, W2, W3 and W4 are N, and the remaining two of W1, W2, W3 and W4 are each CR′. In some such embodiments, W1 and W2 are each N, and W3 and W4 are each CR′. In other such embodiments, W1 and W3 are each N, and W2 and W4 are each CR′. In other such embodiments, W1 and W4 are each N, and W2 and W3 are each CR′. In other such embodiments, W2 and W3 are each N, and W1 and W4 are each CR′. In other such embodiments, W2 and W4 are each N, and W1 and W3 are each CR′. In other such embodiments, W3 and W4 are each N, and W1 and W2 are each CR′.
In yet other embodiments, three of W1, W2, W3 and W4 are N, and the remaining one of W1, W2, W3 and W4 is CR′. In some such embodiments, W1, W2 and W3 are N, and W4 is CR′. In other such embodiments, W1, W2 and W4 are N, and W3 is CR′. In other such embodiments, W1, W3 and W4 are N, and W2 is CR′. In other such embodiments, W2, W3 and W4 are N, and W1 is CR′.
In some embodiments of the compound of Formula (I), Q1 is N and Q2 is CR. In some embodiments of the compound of Formula (I), Q1 is CR and Q2 is N. In some embodiments of the compound of Formula (I), Q1 is N and Q2 is N.
In some embodiments of the compound of Formula (I), each R is independently hydrogen or alkyl. In some such embodiments, each R is independently hydrogen or C1-C4 alkyl. In some embodiments, the C1-C4 alkyl is methyl, ethyl, n-propyl or n-butyl. In some embodiments, the C1-C4 alkyl is methyl or ethyl.
In some embodiments, each R is independently hydrogen or methyl.
In some embodiments of the compound of Formula (I), X1 and X2 are O. In other embodiments, X1 is 0 and X2 is S. In other embodiments, X1 is S and X2 is O. In other embodiments, X1 and X2 are S.
In some embodiments of the compound of Formula (I), n is 0. In other embodiments, n is 1 or 2. In some embodiments, n is 1. In other embodiments, n is 2.
In some embodiments of the compound of Formula (I), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —OR″, —NR″2, or —S(O)2R″. In other embodiments of the compound of Formula (I), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —C(O)R″, —C(O)OR″, —C(O)NH2, —C(O)NHR″, or —C(O)NR″2. In some embodiments of the compound of Formula (I), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —OR″, —NR″2, or —S(O)2R″. In some embodiments of the compound of Formula (I), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, or haloalkenyl. In other embodiments of the compound of Formula (I), L is —OR″, —NR″2, or —S(O)2R″ In some embodiments of the compound of Formula (I), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, or benzyl. In some embodiments of the compound of Formula (I), L is hydrogen, alkyl, alkenyl, or aryl. In some embodiments of the compound of Formula (I), L is hydrogen, alkyl, or alkenyl.
In some embodiments of the compound of Formula (I), L is hydrogen or alkyl. In some embodiments of the compound of Formula (I), L is hydrogen.
In some embodiments, the compound of Formula (I) is:
In some such embodiments, the compound of Formula (I) is:
In other embodiments, the compound of Formula (I) is:
In some such embodiments, the compound of Formula (I) is:
In some embodiments, the compound of Formula (I) is selected from:
In some embodiments, the compound of Formula (I) is selected from:
In some embodiments, the compound of Formula (I) is selected from:
In one embodiment, the compound of Formula (I) is:
In accordance with a second aspect of the invention, there is provided a compound of Formula (IIa), (IIb), or (IIc):
In some embodiments, the compound of Formula (IIa), (IIb), or (IIc) has the structure:
In other embodiments, the compound of Formula (IIa), (IIb), or (IIc) has the structure:
In some embodiments of the compound of Formula (IIa) or (IIc), W1 is N. In some embodiments of the compound of Formula (IIa) or (IIb), W2 is N. In some embodiments of the compound of Formula (IIb) or (IIC), W3 is N.
In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), one of W1, W2 and W3 is N, and the other of W1, W2 and W3 is CRa. In some such embodiments, one of W1, W2 and W3 is N, and the other of W1, W2 and W3 is CH.
In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), W1, W2 and W3 are each CRa.
In some embodiments of the compound of Formula (IIa) or (IIc), W1 is C—NH2, C—NHRb or C—NRb2. In some such embodiments, W1 is C—NH2.
In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), W1, W2 and W3 are each N.
In some embodiments, the compound is of Formula (IIc).
In other embodiments, the compound is of Formula (IIb).
In other embodiments, the compound is of Formula (IIa).
In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), Z is O. In other embodiments of the compound of Formula (IIa), (IIb) or (IIc), Z is S. In other embodiments of the compound of Formula (IIa), (IIb) or (IIc), Z is NH. In other embodiments of the compound of Formula (IIa), (IIb) or (IIc), Z is N-alkyl. In some such embodiments, Z is N-Me.
In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), Q1 is N and Q2 is CR. In some embodiments of the compound of Formula (IIa), (IIb) or (IIc) Q1 is CR and Q2 is N. In some embodiments, Q1 is C—H or C-alkyl. In some such embodiments, Q1 is C-methyl. In other embodiments, Q1 is C—H.
In other embodiments of the compound of Formula (IIa), (IIb) or (IIc) Q1 is N and Q2 is N.
In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), each R is independently hydrogen or alkyl. In some such embodiments, each R is independently hydrogen or C1-C4 alkyl. In some embodiments, C1-C4 alkyl is methyl, ethyl, n-propyl or n-butyl. In some embodiments, C1-C4 alkyl is methyl or ethyl. In some embodiments, each R is independently hydrogen or methyl.
In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), each R′ is independently hydrogen, —NH2, —NHRb or —NRb2. In some such embodiments, each Ra is independently hydrogen or —NH2. In some such embodiments, each Ra is hydrogen.
In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), X1 and X2 are O. In other embodiments, X1 is 0 and X2 is S. In other embodiments, X1 is S and X2 is O. In other embodiments, X1 and X2 are S.
In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), n is 0. In other embodiments, n is 1 or 2. In some embodiments, n is 1. In other embodiments, n is 2.
In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —ORb, —NRb2, or —S(O)2Rb. In other embodiments of the compound of Formula (IIa), (IIb) or (IIc), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —C(O)Rb, —C(O)ORb, —C(O)NH2, —C(O)NHRb, or —C(O)NRb2. In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —ORb, —NRb2, or —S(O)2Rb. In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, or haloalkenyl. In other embodiments of the compound of Formula (IIa), (IIb) or (IIc), L is —ORb, —NRb2, or —S(O)2Rb. In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, or benzyl. In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), L is hydrogen, alkyl, alkenyl, or aryl. In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), L is hydrogen, alkyl, or alkenyl. In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), L is hydrogen or alkyl. In some embodiments of the compound of Formula (IIa), (IIb) or (IIc), L is hydrogen.
In some embodiments, the compound of Formula (IIa), (IIb) or (IIc) is selected from:
In some embodiments, the compound is:
In accordance with a third aspect of the invention, there is provided a compound of Formula (III):
In some embodiments, the compound of Formula (III) has the structure:
In other embodiments, the compound of Formula (III) has the structure:
In some embodiments of the compound of Formula (III), X1 and X2 are O. In other embodiments, X1 is O and X2 is S. In other embodiments, X1 is S and X2 is O. In other embodiments, X1 and X2 are S.
In some embodiments of the compound of Formula (III), n is 0. In other embodiments, n is 1 or 2. In some embodiments, n is 1. In other embodiments, n is 2.
In some embodiments of the compound of Formula (III), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —ORb, —NRb2, or —S(O)2Rb. In other embodiments of the compound of Formula (III), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —C(O)Rb″, —C(O)ORb, —C(O)NH2, —C(O)NHRb, or —C(O)NRb2. In some embodiments of the compound of Formula (III), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —ORb, —NRb2, or —S(O)2Rb. In some embodiments of the compound of Formula (III), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, or haloalkenyl. In other embodiments of the compound of Formula (III), L is —ORb, —NRb2, or —S(O)2Rb. In some embodiments of the compound of Formula (III), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, or benzyl. In some embodiments of the compound of Formula (III), L is hydrogen, alkyl, alkenyl, or aryl. In some embodiments of the compound of Formula (III), L is hydrogen, alkyl, or alkenyl.
In some embodiments of the compound of Formula (III), L is hydrogen or alkyl. In some embodiments of the compound of Formula (III), L is hydrogen.
In accordance with a fourth aspect of the invention, there is provided a compound of formula (IV):
In some embodiments, the compound of Formula (IV) has the structure:
In some embodiments, the compound of Formula (IV) has the structure:
In some embodiments of the compound of Formula (IV), X1 and X2 are O. In other embodiments, X1 is 0 and X2 is S. In other embodiments, X1 is S and X2 is O. In other embodiments, X1 and X2 are S.
In some embodiments of the compound of Formula (IV), one of Q1, Q2, Q3, Q4 and Q5 is N, and the remaining four of Q1, Q2, Q3, Q4 and Q5 are each CR. In some such embodiments, Q1 is N. In other such embodiments, Q2 is N. In other such embodiments, Q3 is N. In other such embodiments, Q4 is N. In other such embodiments, Q5 is N.
In some embodiments of the compound of Formula (IV), two of Q1, Q2, Q3, Q4 and Q5 are N, and the remaining three of Q1, Q2, Q3, Q4 and Q5 are each CR. In some such embodiments, Q1 and Q2 are N, and Q3, Q4 and Q5 are each CR. In other such embodiments, Q2 and Q3 are N, and Q1, Q4 and Q5 are each CR.
In other such embodiments, Q1 and Q3 are N, and Q2, Q4 and Q5 are each CR. In other such embodiments, Q2 and Q4 are N, and Q1, Q3 and Q5 are each CR. In other such embodiments, Q1 and Q4 are N, and Q2, Q3 and Q5 are each CR.
In some embodiments of the compound of Formula (IV), three of Q1, Q2, Q3, Q4 and Q5 are N, and the remaining two of Q1, Q2, Q3, Q4 and Q5 are each CR.
In some embodiments of the compound of Formula (IV), each R is independently hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —NH2, —NHRb, —NRb2, —NRbC(O)Rb, —NRbC(O)ORb, —NO2, —CN, —C(O)Rb, —C(O)ORb, —C(O)NH2, —C(O)NHRb, —C(O)NRb2, —ORb, —OC(O)Rb, —OC(O)ORb, —OC(O)NH2, —OC(O)NHRb, —OC(O)NRb2, —SRb, S(O)2Rb. In some embodiments, each R is independently hydrogen or —NH2. In some embodiments, each R is hydrogen.
In some embodiments of the compound of Formula (IV), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —ORb, —NRb2, or —S(O)2Rb. In other embodiments of the compound of Formula (IV), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —C(O)Rb, —C(O)ORb, —C(O)NH2, —C(O)NHRb, or —C(O)NRb2. In some embodiments of the compound of Formula (IV), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —ORb, —NRb2, or —S(O)2Rb. In some embodiments of the compound of Formula (IV), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, or haloalkenyl. In other embodiments of the compound of Formula (IV), L is —ORb, —NRb2, or —S(O)2Rb. In some embodiments of the compound of Formula (IV), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, or benzyl. In some embodiments of the compound of Formula (IV), L is hydrogen, alkyl, alkenyl, or aryl. In some embodiments of the compound of Formula (IV), L is hydrogen, alkyl, or alkenyl.
In some embodiments of the compound of Formula (IV), L is hydrogen or alkyl. In some embodiments of the compound of Formula (IV), L is hydrogen.
In accordance with a fifth aspect of the invention, there is provided a pharmaceutical composition comprising a compound according to any of the above aspects of the present invention.
The invention also provides a compound according to any of the above aspects of the present invention for use as a cereblon binder.
The invention also provides a compound or composition according to any of the above aspects of the present invention, for use in medicine.
The invention also provides a compound or composition according to any of the above aspects of the present invention, for use in immune-oncology.
The invention also provides a compound or composition according to any of the above aspects of the present invention, for use in the treatment of cancer, autoimmune diseases, macular degeneration (MD) and related disorders, diseases and disorders associated with undesired angiogenesis, skin diseases, pulmonary disorders, asbestos-related disorders, parasitic diseases and disorders, immunodeficiency disorders, atherosclerosis and related conditions, hemoglobinopathy and related disorders, or TNFα related disorders.
The present invention also provides a method for the treatment of cancer, autoimmune diseases, macular degeneration (MD) and related disorders, diseases and disorders associated with undesired angiogenesis, skin diseases, pulmonary disorders, asbestos-related disorders, parasitic diseases and disorders, immunodeficiency disorders, atherosclerosis and related conditions, hemoglobinopathy and related disorders, or TNFα related disorders; wherein the method comprises administering to a patient in need thereof an effective amount of a compound or composition according to any of the above aspects of the present invention.
In some embodiments of the method, the method further comprises administering at least one additional active agent to the patient. In some embodiments, the at least one additional active agent is an anti-cancer agent or an agent for the treatment of an autoimmune disease. In some embodiments, the at least one additional active agent is a small molecule, a peptide, an antibody, a corticosteroid, or a combination thereof. In some embodiments, the at least one additional active agent is at least one of bortezomib, dexamethasone, and rituximab.
The present invention also provides a combined preparation of a compound of any one of the first to fourth aspects of the present invention and at least one additional active agent, for simultaneous, separate or sequential use in therapy.
In some embodiments of the combined preparation, the at least one additional active agent is an anti-cancer agent or an agent for the treatment of an autoimmune disease. In some embodiments, the at least one additional active agent is a peptide, an antibody, a corticosteroid, or a combination thereof. In some embodiments, the at least one additional active agent is at least one of bortezomib, dexamethasone, and rituximab. In some embodiments, the therapy is the treatment of cancer, autoimmune diseases, macular degeneration (MD) and related disorders, diseases and disorders associated with undesired angiogenesis, skin diseases, pulmonary disorders, asbestos-related disorders, parasitic diseases and disorders, immunodeficiency disorders, atherosclerosis and related conditions, hemoglobinopathy and related disorders, or TNFα related disorders.
The present invention also provides a bifunctional compound having the structure:
CLM-[Link]-PTM,
or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate, polymorph or prodrug thereof, wherein:
In some embodiments, [Link] is selected from:
wherein
In some embodiments, [Link] is
In some embodiments, p is an integer from 4 to 11, from 5 to 10, from 6 to 9, or from 7 to 8.
In some embodiments, [Link] is
In some embodiments, [Link] is a bond
In some embodiments, the PTM targets BRD4.
In some embodiments, the PTM is
In some embodiments, at least one of R, R′, Ra, Rb, R1, R2 and R3 is modified so as to include a carboxylic acid group or an ester group.
In some embodiments, the bifunctional compound is selected from
As used herein the term “alkyl” is intended to include both unsubstituted alkyl groups, and alkyl groups which are substituted by one or more additional groups—for example —OH, —OR″, —NH2, —NHR″, —NR″2, —SO2R″, —C(O)R″, —CN, or —NO2. In some embodiments, the alkyl group is an unsubstituted alkyl group. In some embodiments, the alkyl group is a C1-C12 alkyl, a C1-C10 alkyl, a C1-C8 alkyl, a C1-C6 alkyl, or a C1-C4 alkyl group.
As used herein the term “cycloalkyl” is intended to include both unsubstituted cycloalkyl groups, and cycloalkyl groups which are substituted by one or more additional groups—for example —OH, —OR″, —NH2, —NHR″, —NR″2, —SO2R″, —C(O)R″, —CN, or —NO2. In some embodiments, the cycloalkyl group is an unsubstituted alkyl group. In some embodiments, the cycloalkyl group is a cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl group. In some embodiments, the cycloalkyl group is a cyclopentyl or cyclohexyl group. In some embodiments, the cycloalkyl group is a cyclohexyl group.
As used herein the term “alkenyl” is intended to include both unsubstituted alkenyl groups, and alkenyl groups which are substituted by one or more additional groups—for example —OH, —OR″, —NH2, —NHR″, —NR″2, —SO2R″, —C(O)R″, —CN, or —NO2. In some embodiments, the alkenyl group is an unsubstituted alkenyl group. In some embodiments, the alkenyl group is a C2-C12 alkenyl, a C2-C10 alkenyl, a C2-C8 alkenyl, a C2-C6 alkenyl, or a C2-C4 alkenyl group.
As used herein the term “alkynyl” is intended to include both unsubstituted alkynyl groups, and alkynyl groups which are substituted by one or more additional groups—for example —OH, —OR″, halogen, —NH2, —NHR″, —NR″2, —SO2R″, —C(O)R″, —CN, or —NO2. In some embodiments, the alkynyl group is an unsubstituted alkynyl group. In some embodiments, the alkynyl group is a C2-C12 alkynyl, a C2-C10 alkynyl, a C2-C8 alkynyl, a C2-C6 alkynyl, or a C2-C4 alkynyl group.
As used herein the term “aryl” is intended to include both unsubstituted aryl groups, and aryl groups which are substituted by one or more additional groups—for example —OH, —OR″—O-haloalkyl, alkyl, halogen, —NH2, —NHR″, —NR″2, —SO2R″, —C(O)R″, —CN, or —NO2. In some embodiments the aryl group is substituted with one or more additional groups selected from —R″, —O-haloalkyl, alkyl, halogen, —NR″2, —SO2R″, —C(O)R″, —CN, or —NO2. In some embodiments, the aryl group is substituted with one or more additional groups selected from halogen, alkyl and O-haloalkyl. In some embodiments, the aryl group is substituted with one or more additional groups selected from Cl, methyl and O—CF3. In some embodiments, the aryl group is an unsubstituted aryl group. In some embodiments, the aryl group is a C6-C10 aryl, a C6-C8 aryl, or a C6 aryl. As used herein the term “heteroaryl” is intended to include both unsubstituted heteroaryl groups, and heteroaryl groups which are substituted by one or more additional groups—for example —OH, —OR″, halogen, —NH2, —NHR″, —NR″2, —SO2R″, —C(O)R″, —CN, or —NO2. In some embodiments, the heteroaryl group is an unsubstituted heteroaryl group. In some embodiments, the heteroaryl group is a C6-C10 heteroaryl, a C6-C9 heteroaryl, a C6-C8 heteroaryl, or a C6 heteroaryl.
As used herein the term “benzyl” is intended to include both unsubstituted benzyl groups, and benzyl groups which are substituted by one or more additional groups—for example —OH, —OR″, halogen, —NH2, —NHR″, —NR″2, —SO2R″, —C(O)R″, —CN, or —NO2. In some embodiments, the benzyl group is an unsubstituted benzyl group.
As discussed above, the present invention provides compounds of Formulas (I), (IIa)-(IIc), (Ill) and (IV):
Binding of the above compounds to cereblon may alter the specificity of the CRL4CRBN complexes, and induce association of novel substrate proteins, followed by their ubiquitination and degradation. Examples of such proteins include, but are not limited to, IKZF1 and IKZF3.
The above compounds may modulate cereblon in a unique way allowing CRL4CRBN ubiquitin ligase complex to recognise different substrates to those which it would otherwise recognise, and target them for degradation. Consequently, the compounds of the present invention are expected to broaden/modify CRBN's antiproliferative activity, thus extending the range of cancer types sensitive to treatment with CMAs.
The compounds of the present invention are advantageous in terms of their synthetic feasibility. The synthesis of the compounds can be summarized as follows:
One example of a compound of the present invention is 3-(5-amino-2-methylquinolin-3-yl)piperidine-2,6-dione (Compound 1):
3-(5-amino-2-methylquinolin-3-yl)piperidine-2,6-dione (Compound 1) can be synthesised as follows:
wherein Step 1 involves reaction with m-CPBA and phosphoryl bromide; Step 2 involves reaction with 2,6-Bis(benzyloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine, tripotassium phosphate and Pd(dppf)Cl2 CH2Cl2; and Step 3 involves reaction with H2 gas in the presence of Pd on activated charcoal. Full experimental details for the synthesis of 3-(5-amino-2-methylquinolin-3-yl)piperidine-2,6-dione are given in the “Examples” section, below.
Other examples of compounds of the present invention are shown below:
In some embodiments, the compound is
As also discussed in the Examples section, the present inventors have found that various compoundsof the present invention exhibit similar or improved cereblon binding capability to that of the known CMA, CC-122. Despite the pharmaceutical activity of the known CMAs such as CC-122, patients often develop resistance to these compounds. The use of novel compounds—such as those of the present invention, as described above—may help to overcome this clinical obstacle.
One of the serious disadvantages of the currently available CMAs is their safety profile. For example, the teratogenicity of the CMAs is dependent upon the extent to which the CMAs induce degradation of SALL4 transcription factor. Known CMAs induce degradation of several proteins (including SALL4) which bind to CRL4CRBN ligase only in presence of the CMA. SALL4 degradation, observed under treatment with CMAs, is responsible (at least partly) for the teratogenicity of the CMAs. Compounds with diminished capability to induce SALL4 degradation may demonstrate an improved safety profile.
The compounds of the present invention may also possess pharmaceutically advantageous properties, such as increased stability and improved ADMET (absorption, distribution, metabolism, excretion, and/or toxicity) properties.
The compounds of the present invention may be useful in the treatment of various diseases and disorders, including (but not limited to):
The compounds of the present invention may also be useful in preventing, treating, or reducing the risk of developing graft versus host disease (GVHD) or transplant rejection.
The compounds of the present invention may also inhibit the production of certain cytokines including, but not limited to, TNF-α, IL-1β, IL-12, IL-18, GM-CSF, IL-10, TGF-β and/or IL-6. The present compounds may stimulate the production of certain cytokines, and also act as a costimulatory signal for T cell activation, resulting in increased production of cytokines such as, but not limited to, IL-12, IL-2, IL-10, TGF-β and/or IFN-γ. In addition, compounds provided herein can enhance the effects of NK cells and antibody-mediated cellular cytotoxicity (ADCC). Further, compounds provided herein may be immunomodulatory and/or cytotoxic, and thus may be useful as chemotherapeutic agents.
To a solution of appropriate 2-aminobenzaldehyde (1 equiv) in MeOH (0.5-1 M) was added 4-oxopentanoic acid (1 equiv) followed by 2 M NaOH (1.2 equiv). The reaction mixture was refluxed for 18 h, concentrated under reduced pressure and neutralized with acetic acid, the solids were filtered and washed with water, diethyl ether and pentane to give substitutes 2-(2-methylquinolin-3-yl)acetic acid.
To a solution of DCC (1.1 equiv) in DCM were added DMAP (0.8 equiv) and appropriate 2-(quinolin-3-yl)acetic acid (1 equiv) at 0° C. Tert-butanol (3 equiv) was added and the reaction mixture was warmed to RT and stirred for 12 h. The reaction mixture was diluted with water, extracted with ethyl acetate, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography.
To a solution of appropriate tert-butyl 2-(quinolin-3-yl)acetate (1 equiv) in DMF were added K2CO3 (1 equiv), benzyltriethylammonium chloride (1 equiv) and acrylonitrile (1 equiv) and the reaction mixture was stirred at RT for 16 h. The reaction mixture was diluted with water and the product was extracted with ethyl acetate. Combined organic phases were dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography.
To an ice cold solution of appropriate tert-butyl 4-cyano-2-(quinolin-3-yl)butanoate (1 equiv) in DMSO were added H2O2(5 equiv) and K2CO3 (0.1 equiv). The reaction mixture was warmed to RT and stirred for 16 h. The reaction mixture was diluted with water and the product was extracted with ethyl acetate. Combined organic phases were dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography.
In a vial were placed appropriate tert-butyl 5-amino-2-(2-methylquinolin-3-yl)-5-oxopentanoate (1 equiv), p-toluenesulfonic acid (5-10 equiv) and ACN and the reaction mixture was stirred at 80° C. for 2-48 h. The mixture was concentrated under reduced pressure and purified by flash column chromatography or preparative HPLC.
In a vial were placed appropriate nitro compound, 10% Pd/C, ethanol and the reaction mixture was stirred at RT under hydrogen atmosphere (1 bar) until full conversion was achieved. The mixture was filtered through Celite and concentrated under reduced pressure.
In a vial was placed appropriate amino compound, pyridine (0.01-0.1 M) was added followed by appropriate sulfonyl chloride and the reaction mixture was stirred at RT until full conversion was achieved. The mixture was concentrated under reduced pressure and purified by flash column chromatography or preparative HPLC.
To a solution of bromoarene (1 equiv) in dioxane were added KOAc (2 equiv), ((1-(tert-butoxy)vinyl)oxy)(tert-butyl)dimethylsilane (4 equiv) and Pd[P(o-Tol)3]2Cl2 (0.2 equiv) under inert gas and the reaction mixture was stirred at 130° C. for 48 h. The reaction mixture was filtered through Celite, concentrated under reduced pressure and purified by flash column chromatography to give appropriate tert-butyl arylacetate.
Step A: To an ice cold solution of 5-nitro-2-methyl quinoline (2.30 g, 12.22 mmol, 1 equiv) in DCM (25 mL) was added m-CPBA (2.3 g, 13.67 mmol, 1.1 equiv). The reaction mixture was warmed to RT and stirred for 16 h. The mixture was filtered and filtrates were washed with 1 M KOH solution, dried over Na2SO4, and concentrated under reduced pressure to give 2-methyl-5-nitroquinoline 1-oxide (88% yield).
Step B: To an ice cold solution of 2-methyl-5-nitroquinoline 1-oxide (500.0 mg, 2.44 mmol, 1 equiv) in DCM (5 mL) was added POBr3 (1.4 g, 4.9 mmol, 2 equiv) in DCM (5 mL). The reaction mixture was warmed to RT and stirred for 48 h. Ice water was added, the solution was neutralized with 10% NH3 solution, extracted with DCM, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give 2-methyl-3-bromo-5-nitroquinoline (14% yield).
Step C: The reaction was performed according to the general procedure H using 2-methyl-3-bromo-5-nitroquinoline (600 mg, 2.24 mmol, 1 equiv) to give tert-butyl 2-(2-methyl-5-nitroquinolin-3-yl)acetate (58% yield).
Step D: The reaction was performed according to the general procedure C using tert-butyl 2-(2-methyl-5-nitroquinolin-3-yl)acetate (200 mg, 0.662 mmol) to give tert-butyl 4-cyano-2-(2-methyl-5-nitroquinolin-3-yl)butanoate (40% yield).
Step E: The reaction was performed according to the general procedure D using tert-butyl 4-cyano-2-(2-methyl-5-nitroquinolin-3-yl)butanoate (120.0 mg, 0.338 mmol) to give tert-butyl 5-amino-2-(2-methyl-5-nitroquinolin-3-yl)-5-oxopentanoate (51% yield).
Step F: The reaction was performed according to the general procedure E using tert-butyl 5-amino-2-(2-methyl-5-nitroquinolin-3-yl)-5-oxopentanoate (250 mg, 0.670 mmol) to give 3-(2-methyl-5-nitroquinolin-3-yl)piperidine-2,6-dione (69% yield).
1H NMR (500 MHz, DMSO) δ 10.98 (s, 1H), 8.60 (s, 1H), 8.40-8.30 (m, 2H), 7.89 (dd, J=8.5, 7.7 Hz, 1H), 4.42 (dd, J=12.5, 4.7 Hz, 1H), 2.82 (ddd, J=17.8, 12.8, 5.3 Hz, 1H), 2.71 (s, 3H), 2.66-2.61 (m, 1H), 2.44 (dd, J=12.8, 4.3 Hz, 1H), 2.14 (ddt, J=10.0, 7.8, 3.9 Hz, 1H).
LCMS (m/z [M+H]+): 299.9
Step G: The reaction was performed according to the general procedure F using 3-(2-methyl-5-nitroquinolin-3-yl)piperidine-2,6-dione (139 mg, 0.464 mmol) to give 3-(5-amino-2-methylquinolin-3-yl)piperidine-2,6-dione (99% yield).
1H NMR (500 MHz, DMSO) δ 10.91 (s, 1H), 8.26 (s, 1H), 7.42-7.27 (m, 1H), 7.08 (d, J=8.3 Hz, 1H), 6.62 (dd, J=7.6, 0.8 Hz, 1H), 5.85 (s, 2H), 4.22 (dd, J=12.6, 4.8 Hz, 1H), 2.85 (ddd, J=18.1, 13.2, 5.3 Hz, 1H), 2.65 (dt, J=17.0, 3.4 Hz, 1H), 2.60 (s, 3H), 2.46 (dq, J=13.0, 4.1 Hz, 1H), 2.11 (dtd, J=13.0, 5.1, 2.9 Hz, 1H).
LCMS (m/z [M+H]+) 270.0
2-Methyl-5-nitro-8,8a-dihydroquinoline (19.8 g, 105.3 mmol) was dissolved in dichloromethane (250 mL) and cooled to 5° C. in an ice bath. m-CPBA (32.9 g, 133.4 mmol, 70%) was added in portions thereto and the reaction mixture was stirred at room temperature (20-25° C.) for 12 hrs. The mixture was washed with 2M NaOH solution (2×150 mL), dried over anhydrous sodium sulfate, and evaporated under vacuum to afford a yellow solid (22 g). The solid was dissolved in CHCl3 (200 mL), the obtained solution was cooled to 5° C. in the ice-bath, and phosphoryl bromide (62.6 g, 218.3 mmol) in CHCl3 (300 mL) was added dropwise to the reaction mixture. The mixture was stirred at room temperature (20-25° C.) for 12 hrs, poured into ice-water, basified to pH=12 with solid potassium carbonate, and extracted with CHCl3 (3×100 mL). The combined extracts were dried over anhydrous sodium sulfate and evaporated under vacuum. The crude product was purified by flash column chromatography (eluent Hexane-MTBE 0-100%) to afford 2.9 g of 3-bromo-2-methyl-5-nitro-8,8a-dihydroquinoline (10% yield) as a brown solid.
2,6-Bis(benzyloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (4.55 μg, 10.9 mmol), tripotassium phosphate (4.8 g, 22.6 mmol), and Pd(dppf)Cl2 CH2Cl2 (0.86 g, 1 mmol) were added sequentially to a solution of 3-bromo-2-methyl-5-nitro-8,8a-dihydroquinoline (2.9 g, 10.86 mmol) in 1,4-dioxane (50 mL) and water (5 mL). The obtained mixture was stirred at 100° C. for 12 hrs under an argon atmosphere. The solvents were removed under vacuum, the residue was diluted with EtOAc (100 mL) and filtered through a pad of silica gel. The filtrate was evaporated under vacuum and recrystallized from EtOAc to afford 2.05 g 3-[2,6-bis(benzyloxy)pyridin-3-yl]-2-methyl-5-nitro-8,8a-dihydroquinoline (4.3 mmol, 39% yield) as a pale yellow solid.
Pd on activated charcoal (1.2 g) was added to a solution of 3-[2,6-bis(benzyloxy)pyridin-3-yl]-2-methyl-5-nitro-8,8a-dihydroquinoline (2.05 g, 4.29 mmol) in THF/methanol (5:1, 300 mL). The reaction mixture was stirred under H2 atmosphere for 96 hrs. The catalyst was removed by filtration and the filtrate was evaporated under vacuum. The obtained crude product was purified by HPLC (eluent water-acetonitrile) to afford 0.05 g of the target compound 3-(5-amino-2-methylquinolin-3-yl)piperidine-2,6-dione (4% yield) as a white solid.
1H NMR: (500 MHz, DMSO-d6) δ 10.92 (s, 1H), 8.25 (s, 1H), 7.33 (t, J=7.9 Hz, 1H) 7.07 (d, J=8.2 Hz, 1H), 6.61 (d, J=7.5 Hz, 1H), 5.86 (brs, 2H), 4.25-4.17 (m, 1H), 2.89-2.79 (m, 1H), 2.69-2.61 (m, 1H), 2.59 (s, 3H), 2.46-2.36 (m, 1H), 2.15-2.08 (m, 1H)
LCMS (m/z [M+H]+): 270.2
Step A: The reaction was performed according to the general procedure A using 2-amino-6-fluorobenzaldehyde (1.0 g, 7.19 mmol) to give 2-(5-fluoro-2-methylquinolin-3-yl)acetic acid (38% yield).
Step B: The reaction was performed according to the general procedure B using 2-(5-fluoro-2-methylquinolin-3-yl)acetic acid (1.0 g, 4.56 mmol) to give tert-butyl 2-(5-fluoro-2-methylquinolin-3-yl)acetate (35% yield).
Step C: The reaction was performed according to the general procedure C using tert-butyl 2-(5-fluoro-2-methylquinolin-3-yl)acetate (500 mg, 1.81 mmol) to give tert-butyl 4-cyano-2-(5-fluoro-2-methylquinolin-3-yl)butanoate (50% yield).
Step D: The reaction was performed according to the general procedure D using tert-butyl 4-cyano-2-(5-fluoro-2-methylquinolin-3-yl)butanoate (500 mg, 1.52 mmol) to give tert-butyl 5-amino-2-(5-fluoro-2-methylquinolin-3-yl)-5-oxopentanoate (45% yield).
Step E: The reaction was performed according to the general procedure E using 5-amino-2-(5-fluoro-2-methylquinolin-3-yl)-5-oxopentanoate (5.0 mg, 14 μmol) to give 3-(5-fluoro-2-methylquinolin-3-yl)piperidine-2,6-dione (84% yield).
1H NMR (500 MHz, DMSO) δ 10.94 (s, 1H), 8.24 (s, 1H), 7.79 (d, J=8.5 Hz, 1H), 7.70 (td, J=8.2, 6.2 Hz, 1H), 7.37 (dd, J=10.0, 7.6 Hz, 1H), 4.36 (dd, J=12.7, 4.7 Hz, 1H), 2.82 (ddd, J=17.8, 13.2, 5.4 Hz, 1H), 2.68 (s, 3H), 2.61 (dd, J=17.4, 3.5 Hz, 1H), 2.57-2.51 (m, 1H), 2.12 (dtd, J=12.8, 5.1, 2.6 Hz, 1H).
LCMS (m/z [M+H]+): 272.9 Example 3: Synthesis of 3-(5-nitroquinolin-3-yl)piperidine-2,6-dione (Compound 12) and 3-(5-aminoquinolin-3-VI)piperidine-2,6-dione (Compound 14)
Step A: To a solution of 5-nitroquinoline (5.00 g, 28.7 mmol, 1 equiv) in AcOH (140 mL) was added portionwise N-bromosuccinimide (5.11, 43 mmol, 1.5 equiv) and the reaction mixture was refluxed for 16 h. The volatiles were removed under reduced pressure and the residue was neutralized with 6 M NaOH. The product was extracted with DCM, washed with water and brine, concentrated under reduced pressure and purified by flash column chromatography to give 3-bromo-5-nitroquinoline (3.80 g, 52% yield)
Step B: The reaction was performed according to the general procedure H using 3-bromo-5-nitroquinoline (1.00 g, 3.98 mmol, 1 equiv) to give tert-butyl 2-(5-nitroquinolin-3-yl)acetate (69% yield).
Step C: The reaction was performed according to the general procedure C using tert-butyl 2-(5-nitroquinolin-3-yl)acetate (800 mg, 2.78 mmol) to give tert-butyl 4-cyano-2-(5-nitroquinolin-3-yl)butanoate (45% yield).
Step D: The reaction was performed according to the general procedure D using tert-butyl 4-cyano-2-(5-nitroquinolin-3-yl)butanoate (430 mg, 1.257 mmol) to give tert-butyl 5-amino-2-(5-nitroquinolin-3-yl)-5-oxopentanoate (23% yield).
Step E: The reaction was performed according to the general procedure E using tert-butyl 5-amino-2-(5-nitroquinolin-3-yl)-5-oxopentanoate (30 mg, 0.083 mmol) to give 3-(5-nitroquinolin-3-yl)piperidine-2,6-dione (68% yield).
1H NMR (500 MHz, DMSO) δ 11.01 (s, 1H), 9.01 (d, J=2.1 Hz, 1H), 8.72-8.67 (m, 1H), 8.50-8.40 (m, 2H), 7.95 (dd, J=8.4, 7.7 Hz, 1H), 4.32 (dd, J=12.7, 4.8 Hz, 1H), 2.77 (ddd, J=17.4, 12.9, 5.4 Hz, 1H), 2.64-2.59 (m, 1H), 2.48-2.42 (m, 1H), 2.15 (dtd, J=13.0, 5.2, 2.9 Hz, 1H).
LCMS (m/z [M+H]+) 286.0
Step F: The reaction was performed according to the general procedure F using 3-(5-nitroquinolin-3-yl)piperidine-2,6-dione (13.7 mg, 0.048 mmol) to give 3-(5-aminoquinolin-3-yl)piperidine-2,6-dione (27% yield).
1H NMR (500 MHz, DMSO) δ 10.94 (s, 1H), 8.66 (d, J=2.1 Hz, 1H), 8.39 (s, 1H), 7.40 (t, J=8.0 Hz, 1H), 7.18 (d, J=8.2 Hz, 1H), 6.71 (dd, J=7.6, 1.1 Hz, 1H), 5.94 (s, 2H), 4.05 (dd, J=12.6, 4.9 Hz, 1H), 2.79 (ddd, J=17.7, 12.8, 5.3 Hz, 1H), 2.66-2.60 (m, 1H), 2.42-2.35 (m, 1H), 2.14 (dtd, J=13.3, 5.2, 3.1 Hz, 1H).
LCMS (m/z [M+H]+) 256.0
Step A: The reaction was performed according to the general procedure A using 2-amino-5-nitrobenzaldehyde (2.0 g, 12.05 mmol) to give 2-(2-methyl-6-nitroquinolin-3-yl)acetic acid (67% yield).
Step B: The reaction was performed according to the general procedure B using 2-(2-methyl-6-nitroquinolin-3-yl)acetic acid (1.0 g, 4.06 mmol) to give tert-butyl 2-(2-methyl-6-nitroquinolin-3-yl)acetate (40% yield).
Step C: The reaction was performed according to the general procedure C using tert-butyl 2-(2-methyl-6-nitroquinolin-3-yl)acetate (290 mg, 0.96 mmol) to give tert-butyl 4-cyano-2-(2-methyl-6-nitroquinolin-3-yl)butanoate (44% yield).
Step D: The reaction was performed according to the general procedure D using tert-butyl 4-cyano-2-(2-methyl-6-nitroquinolin-3-yl)butanoate (150 mg, 0.423 mmol) to give tert-butyl 5-amino-2-(2-methyl-6-nitroquinolin-3-yl)-5-oxopentanoate (28% yield).
Step E: The reaction was performed according to the general procedure E using tert-butyl 4-cyano-2-(2-methyl-6-nitroquinolin-3-yl)butanoate (30 mg, 0.080 mmol) to give 3-(2-methyl-6-nitroquinolin-3-yl)piperidine-2,6-dione (67% yield).
1H NMR (500 MHz, DMSO) δ 11.00 (s, 1H), 8.96 (d, J=2.6 Hz, 1H), 8.50 (s, 1H), 8.44-8.38 (m, 1H), 8.12 (d, J=9.1 Hz, 1H), 4.39 (dd, J=12.5, 4.7 Hz, 1H), 2.84 (ddd, J=17.6, 12.8, 5.2 Hz, 1H), 2.74 (s, 3H), 2.68-2.62 (m, 1H), 2.44-2.38 (m, 1H), 2.17 (dtd, J=13.1, 5.1, 2.9 Hz, 1H).
LCMS (m/z [M+H]+): 300.0
Step F: The reaction was performed according to the general procedure F using 3-(2-methyl-6-nitroquinolin-3-yl)piperidine-2,6-dione (16 mg, 0.053 mmol) to give 3-(6-amino-2-methylquinolin-3-yl)piperidine-2,6-dione (58% yield).
1H NMR (500 MHz, DMSO) δ 10.88 (s, 1H), 7.73 (s, 1H), 7.61 (d, J=8.9 Hz, 1H), 7.17-7.01 (m, 1H), 6.73 (d, J=2.5 Hz, 1H), 5.49 (s, 2H), 4.17 (dd, J=12.4, 4.8 Hz, 1H), 2.79 (ddd, J=17.7, 12.8, 5.3 Hz, 1H), 2.59 (dt, J=17.1, 3.7 Hz, 1H), 2.54 (s, 3H), 2.40 (dd, J=12.9, 4.3 Hz, 1H), 2.07 (dtd, J=13.2, 5.2, 3.1 Hz, 1H).
LCMS (m/z [M+H]+): 270.05
Step A: The reaction was performed according to the general procedure A using 2-amino-4-nitrobenzaldehyde (560 mg, 3.37 mmol) to give 2-(2-methyl-7-nitroquinolin-3-yl)acetic acid (quantitative).
Step B: The reaction was performed according to the general procedure B using 2-(2-methyl-7-nitroquinolin-3-yl)acetic acid (830 mg, 3.36 mmol) to give tert-butyl 2-(2-methyl-7-nitroquinolin-3-yl)acetate (45% yield).
Step C: The reaction was performed according to the general procedure C using tert-butyl 2-(2-methyl-7-nitroquinolin-3-yl)acetate (460 mg, 1.52 mmol) to give tert-butyl 4-cyano-2-(2-methyl-7-nitroquinolin-3-yl)butanoate (47% yield).
Step D: The reaction was performed according to the general procedure D using tert-butyl 4-cyano-2-(2-methyl-7-nitroquinolin-3-yl)butanoate (255 mg, 0.718 mmol) to give tert-butyl 5-amino-2-(2-methyl-7-nitroquinolin-3-yl)-5-oxopentanoate (35% yield).
Step E: The reaction was performed according to the general procedure E using tert-butyl 4-cyano-2-(2-methyl-7-nitroquinolin-3-yl)butanoate (30 mg, 0.080 mmol) to give 3-(2-methyl-7-nitroquinolin-3-yl)piperidine-2,6-dione (84% yield).
1H NMR (500 MHz, DMSO) δ 10.99 (s, 1H), 8.72 (d, J=2.4 Hz, 1H), 8.37 (s, 1H), 8.28 (dd, J=8.9, 2.3 Hz, 1H), 8.17 (d, J=8.9 Hz, 1H), 4.40 (dd, J=12.7, 4.7 Hz, 1H), 2.84 (ddd, J=18.0, 13.0, 5.3 Hz, 1H), 2.74 (s, 3H), 2.68-2.62 (m, 1H), 2.45 (td, J=12.9, 4.4 Hz, 1H), 2.16 (dtd, J=13.0, 5.2, 2.8 Hz, 1H).
LCMS (m/z [M+H]+): 300.05 Step F: The reaction was performed according to the general procedure F using 3-(2-methyl-7-nitroquinolin-3-yl)piperidine-2,6-dione (18 mg, 0.063 mmol) to give 3-(7-amino-2-methylquinolin-3-yl)piperidine-2,6-dione (95% yield).
1H NMR (500 MHz, DMSO) δ 10.85 (s, 1H), 7.75 (s, 1H), 7.50 (d, J=8.7 Hz, 1H), 6.90 (dd, J=8.7, 2.2 Hz, 1H), 6.84 (d, J=2.2 Hz, 1H), 5.63 (s, 2H), 4.12 (dd, J=12.3, 4.8 Hz, 1H), 2.79 (ddd, J=17.5, 12.7, 5.2 Hz, 1H), 2.60-2.55 (m, 1H), 2.30 (td, J=12.8, 4.2 Hz, 1H), 2.06 (dtd, J=13.2, 5.2, 3.2 Hz, 1H).
LCMS (m/z [M+H]+): 270.0
Step A: The reaction was performed according to the general procedure A using 2-amino-3-nitrobenzaldehyde (5.0 g, 30.04 mmol) to give 2-(2-methyl-8-nitroquinolin-3-yl)acetic acid (40% yield).
Step B: The reaction was performed according to the general procedure B using 2-(2-methyl-8-nitroquinolin-3-yl)acetic acid (3.0 g, 13.36 mmol) to give tert-butyl 2-(2-methyl-8-nitroquinolin-3-yl)acetate (35% yield).
Step C: The reaction was performed according to the general procedure C using tert-butyl 2-(2-methyl-8-nitroquinolin-3-yl)acetate (1.20 g, 3.97 mmol) to give tert-butyl 4-cyano-2-(2-methyl-8-nitroquinolin-3-yl)butanoate (22% yield).
Step D: The reaction was performed according to the general procedure D using tert-butyl 4-cyano-2-(2-methyl-8-nitroquinolin-3-yl)butanoate (310 mg, 0.871 mmol) to give tert-butyl 5-amino-2-(2-methyl-8-nitroquinolin-3-yl)-5-oxopentanoate (46% yield).
Step E: The reaction was performed according to the general procedure E using tert-butyl 4-cyano-2-(2-methyl-8-nitroquinolin-3-yl)butanoate (30 mg, 0.080 mmol) to give 3-(2-methyl-8-nitroquinolin-3-yl)piperidine-2,6-dione (65% yield).
1H NMR (500 MHz, DMSO) δ 10.98 (s, 1H), 8.36 (s, 1H), 8.22-8.13 (m, 2H), 7.72-7.64 (m, 1H), 4.38 (dd, J=12.7, 4.7 Hz, 1H), 2.84 (ddd, J=17.4, 13.1, 5.3 Hz, 1H), 2.68 (s, 3H), 2.64 (dd, J=16.9, 3.7 Hz, 1H), 2.44 (td, J=12.9, 4.2 Hz, 1H), 2.16 (dtd, J=12.8, 5.1, 2.8 Hz, 1H).
LCMS (m/z [M+H]+): 299.95
Step F: The reaction was performed according to the general procedure F using 3-(2-methyl-8-nitroquinolin-3-yl)piperidine-2,6-dione (16 mg, 0.053 mmol) to give 3-(8-amino-2-methylquinolin-3-yl)piperidine-2,6-dione (91% yield).
1H NMR (500 MHz, DMSO) δ 10.90 (s, 1H), 7.92 (s, 1H), 7.20 (t, J=7.7 Hz, 1H), 6.97 (dd, J=8.2, 1.4 Hz, 1H), 6.80 (dd, J=7.5, 1.3 Hz, 1H), 5.78 (s, 2H), 4.24 (dd, J=12.4, 4.8 Hz, 1H), 2.81 (ddd, J=17.8, 12.9, 5.3 Hz, 1H), 2.64 (s, 3H), 2.62-2.57 (m, 1H), 2.44-2.37 (m, 1H), 2.11 (dtd, J=13.1, 5.2, 3.0 Hz, 1H).
LCMS (m/z [M+H]+): 270.0
Step A: The reaction was performed according to the general procedure A using 2-amino-3-chlorobenzaldehyde (1.0 g, 6.42 mmol) to give 2-(8-chloro-2-methylquinolin-3-yl)acetic acid.
Step B: The reaction was performed according to the general procedure B using 2-(8-chloro-2-methylquinolin-3-yl)acetic acid to give tert-butyl 2-(8-chloro-2-methylquinolin-3-yl)acetate (38% yield, two steps).
Step C: The reaction was performed according to the general procedure C using tert-butyl 2-(8-chloro-2-methylquinolin-3-yl)acetate (400 mg, 1.47 mmol) to give tert-butyl 4-cyano-2-(8-chloro-2-methylquinolin-3-yl)butanoate.
Step D: The reaction was performed according to the general procedure D using tert-butyl 4-cyano-2-(8-chloro-2-methylquinolin-3-yl)butanoate to give tert-butyl 5-amino-2-(8-chloro-2-methylquinolin-3-yl)-5-oxopentanoate.
Step E: The reaction was performed according to the general procedure E using tert-butyl 5-amino-2-(8-chloro-2-methylquinolin-3-yl)-5-oxopentanoate to give 3-(8-chloro-2-methylquinolin-3-yl)piperidine-2,6-dione (9% yield, three steps).
1H NMR (400 MHz, DMSO) 10.96 (s, 1H), 8.22 (s, 1H), 7.91-7.84 (m, 2H), 7.50 (t, J=7.8 Hz, 1H), 4.34 (dd, J=12.5, 4.6 Hz, 1H), 2.88-2.77 (m, 1H), 2.71 (s, 3H), 2.66-2.57 (m, 1H), 2.48-2.38 (m, 1H), 2.18-2.12 (m, 1H).
LCMS (m/z [M+H]+): 289.2
Step A: The reaction was performed according to the general procedure A using 2-amino-3-methylbenzaldehyde (1.0 g, 7.39 mmol) to give 2-(2,8-dimethylquinolin-3-yl)acetic acid.
Step B: The reaction was performed according to the general procedure B using 2-(2,8-dimethylquinolin-3-yl)acetic acid to give tert-butyl 2-(2,8-dimethylquinolin-3-yl)acetate (39% yield, two steps).
Step C: The reaction was performed according to the general procedure C using tert-butyl 2-(2,8-dimethylquinolin-3-yl)acetate (500 mg, 1.84 mmol) to give tert-butyl 4-cyano-2-(2,8-dimethylquinolin-3-yl)butanoate (35% yield).
Step D: The reaction was performed according to the general procedure D using tert-butyl 4-cyano-2-(2,8-dimethylquinolin-3-yl)butanoate (200 mg, 0.616 mmol) to give tert-butyl 5-amino-2-(2,8-dimethylquinolin-3-yl)-5-oxopentanoate.
Step E: The reaction was performed according to the general procedure E using tert-butyl 5-amino-2-(2,8-dimethylquinolin-3-yl)-5-oxopentanoate to give 3-(2,8-dimethylquinolin-3-yl)piperidine-2,6-dione (30% yield, two steps).
1H NMR (400 MHz, DMSO) 10.92 (s, 1H), 8.07 (s, 1H), 7.70 (d, J=8.1 Hz, 1H), 7.54 (d, J=6.8 Hz, 1H), 7.40 (dd, J=8.1, 6.8 Hz, 1H), 4.29 (dd, J=12.4, 4.5 Hz, 1H), 2.80-2.65 (m, 1H), 2.69 (s, 3H), 2.67 (s, 3H), 2.65-2.53 (m, 1H), 2.50-2.35 (m, 1H), 2.17-2.08 (m, 1H).
LCMS (m/z [M+H]+): 269.3
Step A: The reaction was performed according to the general procedure A using 2-aminobenzaldehyde (5.0 g, 41.3 mmol) to give 2-(2-methylquinolin-3-yl)acetic acid (17% yield).
Step B: The reaction was performed according to the general procedure B using 2-(2-methylquinolin-3-yl)acetic acid (1.40 g, 6.96 mmol) to give tert-butyl 2-(2-methylquinolin-3-yl)acetate (44% yield).
Step C: The reaction was performed according to the general procedure C using tert-butyl 2-(2-methylquinolin-3-yl)acetate (800 mg, 3.11 mmol) to give tert-butyl 4-cyano-2-(2-methylquinolin-3-yl)butanoate (72% yield).
Step D: The reaction was performed according to the general procedure D using tert-butyl 4-cyano-2-(2-methylquinolin-3-yl)butanoate (700 mg, 2.25 mmol) to give tert-butyl 5-amino-2-(2-methylquinolin-3-yl)-5-oxopentanoate (67% yield).
Step E: The reaction was performed according to the general procedure E using 5-amino-2-(2-methylquinolin-3-yl)-5-oxopentanoate (100 mg, 0.304 mmol) to give 3-(2-methylquinolin-3-yl)piperidine-2,6-dione (76% yield).
1H NMR (500 MHz, DMSO) δ 10.93 (s, 1H), 8.12 (s, 1H), 7.94-7.90 (m, 1H), 7.90-7.85 (m, 1H), 7.69 (ddd, J=8.4, 6.9, 1.5 Hz, 1H), 7.52 (ddd, J=8.1, 6.9, 1.2 Hz, 1H), 4.29 (dd, J=12.5, 4.8 Hz, 1H), 2.83 (ddd, J=17.2, 12.9, 5.3 Hz, 1H), 2.66 (s, 3H), 2.65-2.56 (m, 1H), 2.42 (qd, J=12.9, 4.3 Hz, 1H), 2.13 (dtd, J=13.0, 5.1, 3.0 Hz, 1H).
LCMS (m/z [M+H]+) 255.0
Step A: The reaction was performed according to the general procedure A using 2-amino-4-bromobenzaldehyde (3.00 g, 15.0 mmol) to give 2-(7-bromo-2-methylquinolin-3-yl)acetic acid (45% yield).
Step B: The reaction was performed according to the general procedure B using 2-(7-bromo-2-methylquinolin-3-yl)acetic acid (500 mg, 1.78 mmol) to give tert-butyl 2-(7-bromo-2-methylquinolin-3-yl)acetate (31% yield).
Step C: In a pressure Schlenk flask were placed molybdenum hexacarbonyl (196.3 mg, 0.744 mmol, 1 equiv) and benzyltriethylammonium chloride (169.4 mg, 0.744 mmol, 1 equiv). Dioxane (10 mL) was added and the mixture was heated at 140° C. for 1 h. (S)-1-cyclohexylethylamine (189.2 mg, 1.487 mmol, 2 equiv) and tert-butyl 2-(7-bromo-2-methylquinolin-3-yl)acetate (250.0 mg, 0.744 mmol, 1 equiv) were added, and the reaction was continued at 150° C. for 16 h. The volatiles was removed under reduced pressure and the residue was purified by flash column chromatography to give tert-butyl (S)-2-(7-((1-cyclohexylethyl)carbamoyl)-2-methylquinolin-3-yl)acetate (199.0 mg, 65% yield).
Step D: The reaction was performed according to the general procedure C using tert-butyl (S)-2-(7-((1-cyclohexylethyl)carbamoyl)-2-methylquinolin-3-yl)acetate (150 mg, 0.365 mmol) to give tert-butyl 4-cyano-2-(7-(((S)-1-cyclohexylethyl)carbamoyl)-2-methylquinolin-3-yl)butanoate (77% yield).
Step E: The reaction was performed according to the general procedure D using tert-butyl 4-cyano-2-(7-(((S)-1-cyclohexylethyl)carbamoyl)-2-methylquinolin-3-yl)butanoate (130 mg, 0.280 mmol) to give tert-butyl 5-amino-2-(7-(((S)-1-cyclohexylethyl)carbamoyl)-2-methylquinolin-3-yl)-5-oxopentanoate (59% yield).
Step F: The reaction was performed according to the general procedure E using tert-butyl 5-amino-2-(7-(((S)-1-cyclohexylethyl)carbamoyl)-2-methylquinolin-3-yl)-5-oxopentanoate (75 mg, 0.156 mmol) to give N—((S)-1-cyclohexylethyl)-3-(2,6-dioxopiperidin-3-yl)-2-methylquinoline-7-carboxamide (60% yield).
1H NMR (500 MHz, DMSO) δ 10.97 (s, 1H), 8.49 (s, 1H), 8.41 (d, J=8.6 Hz, 1H), 8.20 (s, 1H), 7.99-7.92 (m, 2H), 4.34 (dd, J=12.5, 4.7 Hz, 1H), 3.92 (h, J=6.9 Hz, 1H), 2.85 (ddd, J=17.7, 12.9, 5.3 Hz, 1H), 2.71 (s, 3H), 2.65 (dt, J=17.2, 3.4 Hz, 1H), 2.46 (qd, J=13.0, 4.3 Hz, 1H), 2.21-2.12 (m, 1H), 1.81 (d, J=12.6 Hz, 2H), 1.74 (d, J=11.2 Hz, 2H), 1.64 (d, J=11.4 Hz, 1H), 1.49 (tdt, J=11.1, 7.0, 3.3 Hz, 1H), 1.31-1.08 (m, 6H), 1.01 (qd, J=12.5, 3.1 Hz, 2H).
LCMS (m/z [M+H]+) 408.1
Step A: The reaction was performed according to the general procedure C using tert-butyl 2-(7-bromo-2-methylquinolin-3-yl)acetate (600 mg, 1.78 mmol) to give tert-butyl 2-(7-bromo-2-methylquinolin-3-yl)-4-cyanobutanoate (31% yield).
Step B: The reaction was performed according to the general procedure D using tert-butyl 2-(7-bromo-2-methylquinolin-3-yl)-4-cyanobutanoate (350 mg, 0.899 mmol) to give tert-butyl 5-amino-2-(7-bromo-2-methylquinolin-3-yl)-5-oxopentanoate.
Step C: The reaction was performed according to the general procedure E using tert-butyl 5-amino-2-(7-bromo-2-methylquinolin-3-yl)-5-oxopentanoate to give 3-(7-bromo-2-methylquinolin-3-yl)piperidine-2,6-dione (67% yield, two steps).
Step D: In a flask were placed 3-(7-bromo-2-methylquinolin-3-yl)piperidine-2,6-dione (80.0 mg, 0.24 mmol, 1 equiv), zinc cyanide (84.6 mg, 0.72 mmol, 3 equiv) and Pd(PPh3)4(27.7 mg, 24 μmol, 0.1 equiv). DMF (2.0 mL) was added and the reaction mixture was stirred at 130° C. for 18 h. The volatiles were removed under reduced pressure and the residue was purified by flash column chromatography to give 3-(2,6-dioxopiperidin-3-yl)-2-methylquinoline-7-carbonitrile (55 mg, 82% yield).
Step E: In a flask were placed 3-(2,6-dioxopiperidin-3-yl)-2-methylquinoline-7-carbonitrile (30.0 mg, 0.107 mmol, 1 equiv), DMF (1.0 mL) and THE (2.0 mL). Raney Nickel (37.8 mg, 0.644 mmol, 6 equiv) was added followed by Boc2O (46.9 mg, 0.215 mmol, 2 equiv) and the reaction mixture was stirred at RT under hydrogen atmosphere (balloon) for 18 h. The reaction mixture was filtered through Celite, solids were washed with EtOH and the filtrates were concentrated under reduced pressure. The crude product was purified by flash column chromatography to give tert-butyl ((3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-7-yl)methyl)carbamate (29 mg, 70% yield).
Step F: In a vial was placed tert-butyl ((3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-7-yl)methyl)carbamate (5.5 mg, 14 μmol, 1 equiv). Dioxane (0.5 mL) was added followed by 12 M HCl (0.1 mL) and the reaction mixture was stirred at RT for 2 h. The volatiles were removed under reduced pressure and the residue was redissolved in DMF (1 mL). DIPEA (0.012 mL, 71 μmol, 5 equiv) was added followed by 3-chloro-4-methylphenylisocyanate (2.9 mg, 17 μmol, 1.2 equiv) and the reaction mixture was stirred at RT for 18 h. The volatiles were removed under reduced pressure and the crude product was purified by preparative HPLC to give 1-(3-chloro-4-methylphenyl)-3-{[3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-7-yl]methyl}urea (4.8 mg, 74% yield).
1H NMR (500 MHz, DMSO) δ 10.94 (s, 1H), 8.78 (s, 1H), 8.10 (s, 1H), 7.86 (d, J=8.4 Hz, 1H), 7.81 (s, 1H), 7.70 (d, J=2.1 Hz, 1H), 7.48 (dd, J=8.4, 1.6 Hz, 1H), 7.21 (d, J=8.4 Hz, 1H), 7.17 (dd, J=8.3, 2.1 Hz, 1H), 6.85 (t, J=6.0 Hz, 1H), 4.51 (d, J=5.9 Hz, 2H), 4.29 (dd, J=12.4, 4.7 Hz, 1H), 2.84 (ddd, J=17.7, 12.9, 5.3 Hz, 1H), 2.66 (s, 3H), 2.65-2.59 (m, 1H), 2.43 (qd, J=13.0, 4.4 Hz, 1H), 2.26 (s, 3H), 2.18-2.10 (m, 1H).
LCMS (m/z [M+H]+) 451.0
In a vial was placed tert-butyl ((3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-7-yl)methyl)carbamate (5.5 mg, 14 μmol, 1 equiv). Dioxane (0.5 mL) was added followed by 12 M HCl (0.1 mL) and the reaction mixture was stirred at RT for 2 h. The volatiles were removed under reduced pressure and the residue was redissolved in DMF (1 mL). 2-(4-Chlorophenyl)-2,2-difluoroacetic acid (4.4 mg, 21.5 μmol, 1.5 equiv) and DIPEA (12 μL, 72 μmol, 5 equiv) were added followed by HATU (8.2 mg, 21 μmol, 1.5 equiv) and the reaction mixture was stirred at RT for 18 h. The volatiles were removed under reduced pressure and the crude product was purified by preparative TLC to give 2-(4-chlorophenyl)-N-{[3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-7-yl]methyl}-2,2-difluoroacetamide (2.9 mg, 42% yield).
1H NMR (500 MHz, DMSO) δ 10.94 (s, 1H), 9.73 (t, J=6.1 Hz, 1H), 8.10 (s, 1H), 7.84 (d, J=8.4 Hz, 1H), 7.70 (s, 1H), 7.69-7.61 (m, 4H), 7.39 (dd, J=8.4, 1.6 Hz, 1H), 4.56 (d, J=6.1 Hz, 2H), 4.29 (dd, J=12.5, 4.7 Hz, 1H), 2.84 (ddd, J=17.7, 12.9, 5.3 Hz, 1H), 2.66 (s, 3H), 2.63 (dt, J=17.2, 3.9 Hz, 1H), 2.42 (qd, J=12.7, 4.0 Hz, 1H), 2.17-2.09 (m, 1H).
LCMS (m/z [M+H]+) 472.0
The reaction was performed according to the general procedure G using 3-(5-amino-2-methylquinolin-3-yl)piperidine-2,6-dione (5.1 mg, 18.9 μmol) to give N-(3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-5-yl)-2-(trifluoromethoxy)benzenesulfonamide (58% yield).
1H NMR (500 MHz, DMSO) δ 10.95 (s, 1H), 10.68 (s, 1H), 8.17 (s, 1H), 7.82 (dd, J=8.1, 1.7 Hz, 1H), 7.74 (s, 1H), 7.71-7.63 (m, 1H), 7.58 (s, 1H), 7.49-7.39 (m, 2H), 7.29 (d, J=6.3 Hz, 1H), 4.26 (dd, J=12.4, 4.7 Hz, 1H), 2.84 (ddd, J=17.7, 12.8, 5.2 Hz, 1H), 2.67 (dt, J=16.9, 3.6 Hz, 1H), 2.63 (s, 3H), 2.24 (qd, J=13.0, 4.3 Hz, 1H), 2.04 (dq, J=8.1, 4.1, 3.1 Hz, 1H).
LCMS (m/z [M+H]+) 494.2
The reaction was performed according to the general procedure G using 3-(6-amino-2-methylquinolin-3-yl)piperidine-2,6-dione (6.5 mg, 24.1 μmol) to give N-(3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-6-yl)-2-(trifluoromethoxy)benzenesulfonamide (65% yield).
1H NMR (500 MHz, DMSO) δ 10.93 (s, 2H), 8.09 (s, 1H), 8.05 (dd, J=7.9, 1.6 Hz, 1H), 7.88-7.81 (m, 1H), 7.75 (ddd, J=8.4, 7.6, 1.7 Hz, 1H), 7.58-7.47 (m, 4H), 4.27 (dd, J=12.6, 4.6 Hz, 1H), 2.82 (ddd, J=17.7, 13.1, 5.3 Hz, 1H), 2.68-2.58 (m, 4H), 2.43 (qd, J=13.0, 4.2 Hz, 1H), 2.14-2.06 (m, 1H).
LCMS (m/z [M+H]+) 494.05
The reaction was performed according to the general procedure G using 3-(7-amino-2-methylquinolin-3-yl)piperidine-2,6-dione (5.3 mg, 19.6 μmol) to give N-(3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-7-yl)-2-(trifluoromethoxy)benzenesulfonaimide (60% yield).
1H NMR (500 MHz, DMSO) δ 10.89 (s, 1H), 8.14 (s, 1H), 8.04 (dd, J=8.1, 1.7 Hz, 1H), 7.96 (s, 1H), 7.77-7.68 (m, 2H), 7.53 (dd, J=8.9, 6.4 Hz, 2H), 7.49 (d, J=2.2 Hz, 1H), 7.30 (dd, J=8.8, 2.3 Hz, 1H), 4.21 (dd, J=12.5, 4.7 Hz, 1H), 2.79 (ddd, J=17.7, 12.8, 5.3 Hz, 1H), 2.60 (dd, J=8.2, 4.5 Hz, 1H), 2.57 (s, 3H), 2.39-2.29 (m, 1H), 2.06 (dtd, J=13.0, 5.1, 3.0 Hz, 1H).
LCMS (m/z [M+H]+) 493.7
The reaction was performed according to the general procedure G using 3-(8-amino-2-methylquinolin-3-yl)piperidine-2,6-dione (5.3 mg, 20 μmol) to give N-(3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-8-yl)-2-(trifluoromethoxy)benzenesulfonamide (63% yield).
1H NMR (500 MHz, DMSO) δ 10.93 (s, 1H), 9.76 (s, 1H), 8.13 (s, 1H), 8.06 (dd, J=7.9, 1.7 Hz, 1H), 7.69 (td, J=7.9, 1.7 Hz, 1H), 7.58 (t, J=9.0 Hz, 2H), 7.52-7.44 (m, 2H), 7.41 (t, J=7.9 Hz, 1H), 4.30 (dd, J=12.6, 4.7 Hz, 1H), 2.81 (ddd, J=17.2, 13.0, 5.3 Hz, 1H), 2.65 (s, 3H), 2.64-2.57 (m, 1H), 2.45-2.34 (m, 1H), 2.10 (dtd, J=12.6, 5.0, 2.7 Hz, 1H).
LCMS (m/z [M+H]+) 493.8
Step A: The reaction was performed according to the general procedure H using 5-bromo-1,6-dimethyl-1H-pyrazolo[3,4-b]pyridine (2.00 g, 8.85 mmol, 1 equiv) to give tert-butyl 2-(1,6-dimethyl-1H-pyrazolo[3,4-b]pyridin-5-yl)acetate (77% yield).
Step B: The reaction was performed according to the general procedure C using tert-butyl 2-(1,6-dimethyl-1H-pyrazolo[3,4-b]pyridin-5-yl)acetate (1.80 g, 6.90 mmol) to give tert-butyl 4-cyano-2-(1,6-dimethyl-1H-pyrazolo[3,4-b]pyridin-5-yl)butanoate (64% yield).
Step C: The reaction was performed according to the general procedure D using tert-butyl 4-cyano-2-(1,6-dimethyl-1H-pyrazolo[3,4-b]pyridin-5-yl)butanoate (700 mg, 2.23 mmol) to give tert-butyl 5-amino-2-(1,6-dimethyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-5-oxopentanoate (48% yield).
Step D: The reaction was performed according to the general procedure E using tert-butyl 5-amino-2-(1,6-dimethyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-5-oxopentanoate (40.0 mg, 0.12 mmol) to give 3-(1,6-dimethyl-1H-pyrazolo[3,4-b]pyridin-5-yl)piperidine-2,6-dione (33% yield).
1H NMR (500 MHz, DMSO) δ 10.88 (s, 1H), 8.01 (s, 1H), 7.98 (s, 1H), 4.22 (dd, J=12.4, 4.8 Hz, 1H), 4.01 (s, 3H), 2.81 (ddd, J=17.3, 12.9, 5.3 Hz, 1H), 2.60 (s, 3H), 2.59-2.55 (m, 1H), 2.41-2.30 (m, 1H), 2.06 (dtd, J=13.1, 5.2, 3.0 Hz, 1H).
LCMS (m/z [M+H]+): 259.1
Step A: The reaction was performed according to the general procedure H using 5-bromothieno[2,3-b]pyridine (1.00 g, 4.67 mmol) to give tert-butyl 2-(thieno[2,3-b]pyridin-5-yl)acetate (51% yield).
Step B: The reaction was performed according to the general procedure C using tert-butyl 2-(thieno[2,3-b]pyridin-5-yl)acetate (500 mg, 2.00 mmol) to give tert-butyl 4-cyano-2-(thieno[2,3-b]pyridin-5-yl)butanoate (41% yield).
Step C: The reaction was performed according to the general procedure D using tert-butyl 4-cyano-2-(thieno[2,3-b]pyridin-5-yl)butanoate (200 mg, 0.632 mmol) to give tert-butyl 5-amino-5-oxo-2-(thieno[2,3-b]pyridin-5-yl)pentanoate.
Step D: The reaction was performed according to the general procedure E using tert-butyl 5-amino-5-oxo-2-(thieno[2,3-b]pyridin-5-yl)pentanoate to give 3-(thieno[2,3-b]pyridin-5-yl)piperidine-2,6-dione (20% yield, two steps).
1H NMR (400 MHz, DMSO) δ 10.90 (s, 1H), 8.46 (d, J=2.0 Hz, 1H), 8.15 (d, J=2.0 Hz, 1H), 7.88 (d, J=5.9 Hz, 1H), 7.43 (d, J=5.9 Hz, 1H), 4.09 (dd, J=12.4, 4.8 Hz, 1H), 2.80-2.70 (m, 1H), 2.63-2.52 (m, 1H), 2.41-2.31 (m, 1H), 2.15-2.06 (m, 1H).
LCMS (m/z [M+H]+) 247.2
Step A: The reaction was performed according to the general procedure A using 2-amino-4-methoxybenzaldehyde (600 mg, 3.96 mmol) to give 2-(7-methoxy-2-methylquinolin-3-yl)acetic acid (43% yield).
Step B: The reaction was performed according to the general procedure B using 2-(7-methoxy-2-methylquinolin-3-yl)acetic acid (400 mg, 1.72 mmol) to give tert-butyl 2-(7-methoxy-2-methylquinolin-3-yl)acetate (26% yield).
Step C: The reaction was performed according to the general procedure C using tert-butyl 2-(7-methoxy-2-methylquinolin-3-yl)acetate (130 mg, 0.452 mmol) to give tert-butyl 4-cyano-2-(7-methoxy-2-methylquinolin-3-yl)butanoate (75% yield).
Step D: The reaction was performed according to the general procedure D using tert-butyl 4-cyano-2-(7-methoxy-2-methylquinolin-3-yl)butanoate (100 mg, 0.293 mmol) to give tert-butyl 5-amino-2-(7-methoxy-2-methylquinolin-3-yl)-5-oxopentanoate.
Step E: The reaction was performed according to the general procedure E using 5-amino-2-(7-methoxy-2-methylquinolin-3-yl)-5-oxopentanoate to give 3-(7-methoxy-2-methylquinolin-3-yl)piperidine-2,6-dione (28% yield, two steps).
1H NMR (400 MHz, DMSO) δ 10.90 (s, 1H), 8.01 (s, 1H), 7.77 (d, J=8.9 Hz, 1H), 7.31 (d, J=2.1 Hz, 1H), 7.16 (dd, J=8.9, 2.2 Hz, 1H), 4.24 (dd, J=12.4, 4.8 Hz, 1H), 3.90 (s, 3H), 2.87-2.75 (m, 1H), 2.62 (s, 3H), 2.43-2.31 (m, 1H), 2.17-2.05 (m, 1H).
LCMS (m/z [M+H]+) 284.8
Step A: To a solution of tert-butyl 2-(2-methyl-8-nitroquinolin-3-yl)acetate (1.00 g, 3.30 mmol) in THE (20 mL) at −78° C. was added LDA (1M in THF, 7.26 mL, 7.26 mmol, 2.2 equiv). The solution was stirred for 30 min and bromoacetonitrile (0.920 mL, 13.2 mmol, 4 equiv) was added dropwise. The solution was warmed to RT and stirred for 12 h. The reaction mixture quenched with 1M HCl and the product was extracted with ethyl acetate. Combined organic phases were dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give tert-butyl 3-cyano-2-(2-methyl-8-nitroquinolin-3-yl)propanoate (20% yield).
Step B: The reaction was performed according to the general procedure D using tert-butyl 3-cyano-2-(2-methyl-8-nitroquinolin-3-yl)propanoate (200 mg, 0.585 mmol) to give tert-butyl 4-amino-2-(2-methyl-8-nitroquinolin-3-yl)-4-oxobutanoate.
Step C: The reaction was performed according to the general procedure E using tert-butyl 4-amino-2-(2-methyl-8-nitroquinolin-3-yl)-4-oxobutanoate to give 3-(2-methyl-8-nitroquinolin-3-yl)pyrrolidine-2,5-dione (20% yield, two steps).
Step D: The reaction was performed according to the general procedure F using 3-(2-methyl-8-nitroquinolin-3-yl)pyrrolidine-2,5-dione (25 mg, 0.087 mmol) to give 3-(8-amino-2-methylquinolin-3-yl)pyrrolidine-2,5-dione (84% yield).
1H NMR (400 MHz, DMSO) δ 11.44 (s, 1H), 7.97 (s, 1H), 7.21 (dd, J=8.0, 7.5 Hz, 1H), 6.98 (d, J=8.0 Hz, 1H), 6.80 (d, J=7.5 Hz, 1H), 5.78 (s, 1H), 4.54 (dd, J=9.6, 6.0 Hz, 1H), 3.21 (dd, J=18.0, 9.6 Hz, 1H), 2.81 (dd, J=18.0, 6.0 Hz, 1H), 2.67 (s, 3H).
LCMS (m/z [M+H]+): 256.1
Step A: In a flask were placed 3-(7-amino-2-methylquinolin-3-yl)piperidine-2,6-dione (71 mg, 0.265 mmol), tetrabutylammonium iodide (97.9 mg, 0.265 mmol, 1 equiv) and DMF (15 mL). DIPEA (185 μL, 1.02 mmol, 4 equiv) was added followed by tert-butyl bromoacetate (51.7 mg, 0.265 mmol, 1 equiv) and the reaction mixture was stirred at 60° C. for 4 h. The reaction mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography to give tert-butyl (3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-7-yl)glycinate (14% yield).
Step B: In a vial was placed tert-butyl (3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-7-yl)glycinate (14.0 mg, 0.037 mmol). Dioxane (1 mL) was added followed by 12M HCl (2 mL). The reaction mixture was stirred at RT for 1 h and concentrated under reduced pressure to give (3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-7-yl)glycine (quant.).
Step C: In a vial were placed (3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-7-yl)glycine (6.2 mg, 0.0.19 mmol), (S)—N-(8-aminooctyl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide (12.0 mg, 0.023 mmol, 1.2 equiv). DMF (1 mL) was added followed by DIPEA (26 μL, 0.152 mmol, 8 equiv) and HATU (8.7 mg, 0.023 mmol, 1.2 equiv), and the reaction mixture was stirred at RT for 2 h. The volatiles were removed under reduced pressure and the residue was purified by preparative HPLC to give 2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-N-(8-(2-((3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-7-yl)amino)acetamido)octyl)acetamide (35% yield).
1H NMR (500 MHz, DMSO) δ 10.85 (s, 1H), 8.12 (t, J=5.7 Hz, 1H), 7.94 (t, J=5.7 Hz, 1H), 7.78 (s, 1H), 7.54 (d, J=8.8 Hz, 1H), 7.47 (d, 2H), 7.42 (d, 2H), 7.01 (dd, J=8.9, 2.3 Hz, 1H), 6.65 (d, J=2.2 Hz, 1H), 6.48 (t, J=5.9 Hz, 1H), 4.50 (dd, J=8.0, 6.1 Hz, 1H), 4.14 (dd, J=12.2, 4.8 Hz, 1H), 3.73 (d, J=5.8 Hz, 2H), 3.30-3.13 (m, 2H), 3.12-3.03 (m, 4H), 2.78 (ddd, J=17.5, 12.7, 5.3 Hz, 1H), 2.59 (s, 3H), 2.57-2.54 (m, 1H), 2.53 (s, 3H), 2.40 (s, 3H), 2.37-2.25 (m, 1H), 2.10-2.01 (m, 1H), 1.62 (s, 3H), 1.44-1.34 (m, 4H), 1.32-1.14 (m, 8H).
LCMS (m/z [M+H]+): 836.3
Step A: In a flask were placed 3-(6-amino-2-methylquinolin-3-yl)piperidine-2,6-dione (18.0 mg, 0.067 mmol), tetrabutylammonium iodide (24.7 mg, 0.067 mmol, 1 equiv) and DMF (1 mL). DIPEA (47 μL, 0.268 mmol, 4 equiv) was added followed by tert-butyl bromoacetate (13.1 mg, 0.067 mmol, 1 equiv) and the reaction mixture was stirred at 60° C. for 3 h. Second portion of tert-butyl bromoacetate (13.1 mg, 0.067 mmol, 1 equiv) was added and heating was continued for another 3 h. The reaction mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography to give tert-butyl (3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-6-yl)glycinate (47% yield).
Step B: In a vial was placed tert-butyl (3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-6-yl)glycinate (12.3 mg, 0.032 mmol). Dioxane (2 mL) was added followed by 12M HCl (3 mL). The reaction mixture was stirred at RT for 1 h and concentrated under reduced pressure to give (3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-6-yl)glycine (quant.).
Step C: In a vial were placed (3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-6-yl)glycine (10.5 mg, 0.032 mmol) and (S)—N-(8-aminooctyl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide (18.1 mg, 0.032 mmol, 1 equiv). DMF (3 mL) was added followed by DIPEA (56 μL, 0.320 mmol, 10 equiv) and HATU (14.6 mg, 0.038 mmol, 1.2 equiv), and the reaction mixture was stirred at RT for 6 h. The volatiles were removed under reduced pressure and the residue was purified by preparative HPLC to give 2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-N-(8-(2-((3-(2,6-dioxopiperidin-3-yl)-2-methylquinolin-6-yl)amino)acetamido)octyl)acetamide (40% yield).
1H NMR (500 MHz, DMSO) δ 10.88 (s, 1H), 8.14 (t, J=5.7 Hz, 1H), 7.92 (t, J=5.8 Hz, 1H), 7.71 (s, 1H), 7.63 (d, J=9.0 Hz, 1H), 7.47 (d, J=8.7 Hz, 2H), 7.42 (d, J=8.7 Hz, 2H), 7.19 (dd, J=9.0, 2.5 Hz, 1H), 6.52 (d, J=2.5 Hz, 1H), 6.35 (t, J=5.8 Hz, 1H), 4.50 (dd, J=8.1, 6.0 Hz, 1H), 4.17 (dd, J=12.3, 4.8 Hz, 1H), 3.70 (d, J=5.7 Hz, 2H), 3.29-3.13 (m, 2H), 3.12-3.00 (m, 4H), 2.84-2.70 (m, 1H), 2.58 (s, 3H), 2.57-2.54 (m, 1H), 2.53 (s, 3H), 2.40 (s, 3H), 2.38-2.30 (m, 1H), 2.11-2.02 (m, 1H), 1.61 (s, 3H), 1.43-1.26 (m, 4H), 1.28-1.12 (m, 8H).
LCMS (m/z [M+H]+): 836.4
CRBN-DDB1 protein complex was mixed with Cy5-labelled thalidomide and a compound to be tested (the “test compound”). The test solution contained 50 mM Tris pH=7.0, 200 mM NaCl, 0.02% v/v Tween-20, 2 mM DTT, 5 nM Cy5-labelled thalidomide (the tracer), 25 nM CRBN-DDB1 protein, 2% v/v DMSO. The test solution was added to a 384-well assay plate.
The plate was spun-down (1 min, 1000 rpm, 22° C.) and then shaken using a VibroTurbulator for 10 min at room temperature (20-25° C.), with the frequency set to level 3. The assay plate with protein and the tracer was incubated for 60 min at room temperature (20-25° C.) prior to read-out with a plate reader. Read-out (fluorescence polarization) was performed by a Pherastar plate reader, using a Cy5 FP Filterset (590 nm/675 nm).
The FP experiment was carried out with various concentrations of the test compounds in order to measure Ki values.
The Ki values of competitive inhibitors were calculated using the equation based on the IC50 values of relationship between compound concentration and measured fluorescence polarization, the Kd value of the Cy5-T and CRBN/DDB1 complex, and the concentrations of the protein and the tracer in the displacement assay (as described by Z. Nikolovska-Coleska et al., Analytical Biochemistry 332 (2004) 261-273).
Compounds are categorized based on their activity to CRBN defined as Ki. As reported in Table 1, below, the compounds of the present invention interact with CRBN-DDB1 protein within similar affinity range as reported for reference compounds.
As can be seen from Table 1, above, the compounds of the present invention exhibited similar CRBN binding affinity (Ki in the same concentration range) as the reference compounds.
The effect of various compounds of the invention and various reference compounds on SALL4 degradation in the Kelly cell line was investigated, using the degradation assay protocol below.
Kelly cells were maintained in RPMI-1640 medium, supplemented with penicillin/streptomycin and 10% Fetal Bovine Serum (FBS). Cells were seeded on 6-well plates, and the compounds to be tested were added at the desired concentration range. Final DMSO concentration was 0.25%. After 24 h incubation (37° C., 5% CO2), cells were washed and cell lysates were prepared using RIPA lysis buffer. The amount of protein was determined via BCA assay, and the appropriate quantity was then loaded on the precast gel for the protein separation. After primary and secondary antibody staining, the membranes were washed and signals developed. The densitometry analysis was implemented to obtain the numeric values used later in the protein level evaluation process.
The compounds tested in this assay were Thalidomide, CC-122 and compound 1 of the present invention, at concentrations of 1-20 μM for 24 h. The results are shown in
As illustrated in
The effect of various compounds of the invention and various reference compounds on IKZF1 degradation in the H929 cell line was investigated, using the degradation assay protocol below.
H929 cells were maintained in RPMI-1640 medium, supplemented with penicillin/streptomycin, 10% Fetal Bovine Serum (FBS) and 0.05 mM 2-Mercaptoethanol. Cells were seeded on 6- or 12-well plates, and the compounds to be tested were added at the desired concentration range. Final DMSO concentration was 0.25%. After 24 h incubation (37° C., 5% CO2), cells were harvested, washed and cell lysates were prepared using RIPA lysis buffer. The amount of protein was determined via BCA assay, and the appropriate quantity was then loaded on the precast gel for the protein separation. After primary and secondary Ab staining, the membranes were washed and signals developed. The densitometry analysis was implemented to obtain the numeric values used later in the protein level evaluation process.
The compounds tested in this assay were Thalidomide, and compound 1 of the present invention, at concentrations of 1-20 μM for 24 h. The results are shown in
As illustrated in
The effect of various compounds of the invention and various reference compounds on IKZF3 degradation in the H929 cell line was investigated, using the degradation assay protocol below.
H929 cells were maintained in RPMI-1640 medium, supplemented with penicillin/streptomycin and 10% Fetal Bovine Serum (FBS) and 0.05 mM 2-Mercaptoethanol. Cells were seeded on 6- or 12-well plates, and the compounds to be tested were added at the desired concentration range. Final DMSO concentration was 0.25%. After 24 h incubation (37° C., 5% CO2), cells were harvested, washed and cell lysates were prepared using RIPA lysis buffer. The amount of protein was determined via BCA assay, and the appropriate quantity was then loaded on the precast gel for the protein separation. After primary and secondary Ab staining, the membranes were washed and signals developed. The densitometry analysis was implemented to obtain the numeric values used later in the protein level evaluation process.
The compounds tested in this assay were Thalidomide, and compound 1 of the present invention, at concentrations of 1-20 μM for 24 h. The results are shown in
As illustrated in
The effect of compound 1 of the invention on the viability of H929 (myeloma) was investigated, using the CTG assay protocol below.
Three thousand cells in 50 μL of culture medium were plated in 384-well plate, and incubated with 50, 13, and 2 μM of each compound for 72 hours. ATP content in the remaining cells after the treatment was quantitated with the CellTiter-Glo Luminescent Viability Assay Kit (Promega). The activity of the compound at each concentration was shown as percentage viability; 100% viability was the ATP content in the cells incubated with DMSO, the carrier of the compounds. The results are presented in Table 5.
As can be seen from Table 5, the compounds of the invention may be useful in the treatment of cancer.
The stability of various compounds of the present invention over a period of 48 hours' incubation at 37° C. in phosphate-buffered saline (PBS)/10% Fetal Bovine Serum (FBS) was analysed by liquid chromatography-mass spectrometry (LC-MS). The results are shown in
Aliquots of the compounds in DMSO (20 mM) were diluted in phosphate-buffered saline (PBS) with 10% Fetal Bovine Serum (FBS) to give the concentration of 0.5 mM. Samples were incubated at 37° C. Samples for LC-MS analyses were taken at the beginning of incubation (0 hours) and after 2, 4, 6, 8, 10, 24, 34, and 48 hours.
For the LC-MS analyses 30 μL of samples were taken, thoroughly mixed with 30 μL of acetonitrile, and vortexed. The samples were then centrifuged (10° C., 10 min, 15 000 xg). Supernatants were transferred to the HPLC vials. For Compounds 1, 15 and 18, supernatants were additionally diluted twice with water prior to analysis.
For Compound 4, Compound 15, Compound 18, and Lenalidomide Kinetex XB—C18 2.6 μm, 50×2.1 mm column kept at 40° C. LC-MS grade mobile phases of water+0.1% formic acid (A) and acetonitrile+0.1% formic acid (B). Elution gradient (flow 0.5 mL/min): 0 min 5% B, 4 min 95% B, 5 min 95% B, 5.2 min 5% B, 7 min 5% B.
Shim-pack Scepter C18-120 3 μm, 150×3 mm column kept in 40° C. LC-MS grade mobile phases of water+0.1% formic acid (A) and acetonitrile+0.1% formic acid (B). Elution gradient (flow 0.5 mL/min): 0 min 5% B, 15 min 95% B, 18 min 95% B, 19 min 5% B, 25 min 5% B.
Chromatograms were integrated and areas of observed peaks were calculated. Different wavelengths were used for quantification of the compounds: Compound 4 (306±4 nm), Compound 15 (244±4 nm), Compound 18 (252±4 nm), Lenalidomide (220±4 nm), CC-122 (235±4 nm), Compound 1 (271±4 nm).
As illustrated in
The effect of bifunctional compounds of the invention on BRD4 degradation in the H929 cell line can be investigated, using the degradation assay protocol below.
H929 cells are maintained in RPMI-1640 medium (ATCC modified, cat.: Gibco A1049101), supplemented with penicillin/streptomycin, 10% Fetal Bovine Serum (FBS) and 0.05 mM 2-Mercaptoethanol. Cells are seeded on 6-well plates (1×10{circumflex over ( )}6 cells/condition) and the compounds to be tested are added at the desired concentration range. Final DMSO concentration is 0.25%. After 6 h incubation (37° C., 5% CO2), cells are harvested and washed. Next, the cell lysates are prepared using RIPA lysis buffer. The amount of protein is determined via BCA assay, and the appropriate quantity is then loaded on pre-filled microplate. The analysis is performed using SIMPLE WESTERN™ technology (from Protein Simple), which is an automated, capillary-based immunoassay. The numeric values for the further protein level evaluation process are counted using the software dedicated for Simple Western analysis. Protein normalization is based on the Protein Normalization Reagent by Protein Simple. Numeric values are presented as % of DMSO control, using the following labels:
Bifunctional compounds of the invention induce degradation of BRD4 protein.
A list of the abbreviations used in the present application is shown in Table 6, below:
As used herein, the term “room temperature” means a temperature of between 20° C. and 25° C.
As used herein, the term “small molecule” means an organic compound with a molecular weight of less than 900 Daltons.
Further embodiments are described below with reference to the following numbered clauses:
Clause 1. A compound of Formula (I):
Clause 2. The compound of clause 1, having the structure:
Clause 3. The compound of clause 1, having the structure:
Clause 4. The compound of any preceding clause, wherein one of W1, W2, W3 and W4 is N, and the remaining three of W1, W2, W3 and W4 are each CR′.
Clause 5. The compound of clause 4, wherein W1 is N, and W2, W3 and W4 are CR′.
Clause 6. The compound of clause 4, wherein W2 is N, and W1, W3 and W4 are CR′.
Clause 7. The compound of clause 4, wherein W3 is N, and W1, W2 and W4 are CR′.
Clause 8. The compound of clause 4, wherein W4 is N, and W1, W2 and W3 are CR′.
Clause 9. The compound of any one of clauses 1-3, wherein W1, W2, W3 and W4 are each CR′.
Clause 10. The compound of clause 9, wherein W2, W3 and W4 are each CH.
Clause 11. The compound of clause 9 or clause 10, wherein W1 is C—NH2, C—NHR″ or C—NR″2; optionally C—NH2.
Clause 12. The compound of any one of clauses 1-3, wherein two of W1, W2, W3 and W4 are N, and the remaining two of W1, W2, W3 and W4 are each CR′.
Clause 13. The compound of any one of clauses 1-3, wherein three of W1, W2, W3 and W4 are N, and the remaining one of W1, W2, W3 and W4 is CR′.
Clause 14. The compound of any preceding clause, wherein L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —OR″, —NR″2, or —S(O)2R″; optionally wherein L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, or benzyl; further optionally wherein L is hydrogen.
Clause 15. The compound of any one of clauses 1-3, wherein the compound is:
Clause 16. The compound of any one of clauses 1-3, wherein the compound is:
Clause 17. The compound of clause 16, wherein the compound is:
Clause 18. A compound of Formula (IIa), (IIb), or (IIc):
Clause 19. The compound of clause 18, having the structure:
Clause 20. The compound of clause 18, having the structure:
Clause 21. The compound of any one of clauses 18-20, wherein W1 is N.
Clause 22. The compound of any one of clauses 18-21, wherein W2 is N.
Clause 23. The compound of any one of clauses 18-22, wherein W3 is N.
Clause 24. The compound of any one of clauses 18-23, wherein one of W1, W2 and W3 is N, and the other of W1, W2 and W3 is CR′.
Clause 25. The compound of clause 24, wherein one of W1, W2 and W3 is N, and the other of W1, W2 and W3 is CH.
Clause 26. The compound of any one of clauses 18-25, wherein W1, W2 and W3 are each CR′.
Clause 27. The compound of any one of clauses 18-25, wherein W1 is C—NH2, C—NHR″ or C—NR″2; optionally C—NH2.
Clause 28. The compound of any one of clauses 18-23, wherein W1, W2 and W3 are each N.
Clause 29. The compound of any one of clauses 18-28, wherein Z is O.
Clause 30. The compound of any one of clauses 18-28, wherein Z is S.
Clause 31. The compound of any one of clauses 18-28, wherein Z is NH.
Clause 32. The compound of any preceding clause, wherein Q1 is N and Q2 is CR.
Clause 33. The compound of any one of clauses 1-31, wherein Q1 is CR and Q2 is N.
Clause 34. The compound of any one of clauses 1-31, wherein Q1 is N and Q2 is N.
Clause 35. The compound of any preceding clause, wherein each R is independently hydrogen or alkyl; optionally hydrogen or C1-C4 alkyl; further optionally wherein the C1-C4 alkyl is methyl or ethyl; further optionally wherein each R is independently hydrogen or methyl.
Clause 36. The compound of any preceding clause, wherein each R′ is independently hydrogen, —NH2, —NHR″ or —NR″2; optionally hydrogen or —NH2.
Clause 37. A compound of Formula (III):
Clause 38. The compound of clause 37, having the structure:
Clause 39. The compound of clause 37, having the structure:
Clause 40. The compound of any preceding clause, wherein X1 and X2 are O.
Clause 41. The compound of any one of clauses 1-39, wherein X1 is O and X2 is S.
Clause 42. The compound of any one of clauses 1-39, wherein X1 is S and X2 is O.
Clause 43. The compound of any one of clauses 1-39, wherein X1 and X2 are S.
Clause 44. The compound of any preceding clause, wherein n is 0.
Clause 45. The compound of any one of clauses 1-43, wherein n is 1 or 2.
Clause 46. The compound of any one of clauses 1-43, wherein n is 1.
Clause 47. The compound of any one of clauses 1-43, wherein n is 2.
Clause 48. A compound of formula (IV):
Clause 49. The compound of clause 48, having the structure:
Clause 50. The compound of clause 48, having the structure:
Clause 51. The compound of any one of clauses 48-50, wherein X1 and X2 are O.
Clause 52. The compound of any one of clauses 48-50, wherein X1 is O and X2 is S.
Clause 53. The compound of any one of clauses 48-50, wherein X1 is S and X2 is O.
Clause 54. The compound of any one of clauses 48-50, wherein X1 and X2 are S.
Clause 55. The compound of any one of clauses 48-54, wherein one of Q1, Q2, Q3, Q4 and Q5 is N, and the remaining four of Q1, Q2, Q3, Q4 and Q5 are each CR.
Clause 56. The compound of clause 55, wherein Q1 is N.
Clause 57. The compound of clause 55, wherein Q2 is N.
Clause 58. The compound of clause 55, wherein Q3 is N.
Clause 59. The compound of any one of clauses 48-54, wherein two of Q1, Q2, Q3, Q4 and Q5 are N, and the remaining three of Q1, Q2, Q3, Q4 and Q5 are each CR.
Clause 60. The compound of clause 59, wherein Q1 and Q2 are N, and Q3, Q4 and Q5 are each CR.
Clause 61. The compound of clause 59, wherein Q2 and Q3 are N, and Q1, Q4 and Q5 are each CR.
Clause 62. The compound of clause 59, wherein Q1 and Q3 are N, and Q2, Q4 and Q5 are each CR.
Clause 63. The compound of clause 59, wherein Q2 and Q4 are N, and Q1, Q3 and Q5 are each CR.
Clause 64. The compound of clause 59, wherein Q1 and Q4 are N, and Q2, Q3 and Q5 are each CR.
Clause 65. The compound of any one of clauses 48-54, wherein three of Q1, Q2, Q3, Q4 and Q5 are N, and the remaining two of Q1, Q2, Q3, Q4 and Q5 are each CR.
Clause 66. The compound of any one of clauses 48-65, wherein each R is independently hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —NH2, —NHR″, —NR″2, —NR″C(O)R″, —NR″C(O)OR″, —NO2, —CN, —C(O)R″, —C(O)OR″, —C(O)NH2, —C(O)NHR″, —C(O)NR″2, —OR″, —OC(O)R″, —OC(O)OR″, —OC(O)NH2, —OC(O)NHR″, —OC(O)NR″2, —SR″, S(O)2R″; optionally wherein each R is hydrogen or alkyl, further optionally wherein each R is hydrogen.
Clause 67. The compound of any one of clauses 18-66, wherein L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —OR″, —NR″2, or —S(O)2R″; optionally wherein L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, or benzyl.
Clause 68. The compound of clause 67, wherein L is hydrogen.
Clause 69. A compound of any one of the preceding clauses, for use as a cereblon binder.
Clause 70. A pharmaceutical composition comprising a compound of any one of clauses 1-68.
Clause 71. A compound of any one of clauses 1-68, or a composition according to clause 70, for use in medicine.
Clause 72. A compound of any one of clauses 1-68, or a composition according to clause 70, for use in immune-oncology.
Clause 73. A compound of any one of clauses 1-68, or a composition according to clause 70, for use in the treatment of cancer, autoimmune diseases, macular degeneration (MD) and related disorders, diseases and disorders associated with undesired angiogenesis, skin diseases, pulmonary disorders, asbestos-related disorders, parasitic diseases and disorders, immunodeficiency disorders, atherosclerosis and related conditions, hemoglobinopathy and related disorders, or TNFα related disorders.
Clause 74. A method for the treatment of cancer, autoimmune diseases, macular degeneration (MD) and related disorders, diseases and disorders associated with undesired angiogenesis, skin diseases, pulmonary disorders, asbestos-related disorders, parasitic diseases and disorders, immunodeficiency disorders, atherosclerosis and related conditions, hemoglobinopathy and related disorders, or TNFα related disorders;
Clause 75. The method of clause 74, further comprising administering at least one additional active agent to the patient.
Clause 76. A combined preparation of a compound of any one of clauses 1-68 and at least one additional active agent, for simultaneous, separate or sequential use in therapy.
Clause 77. The combined preparation of clause 76, or the method of clause 75, wherein the at least one additional active agent is an anti-cancer agent or an agent for the treatment of an autoimmune disease.
Clause 78. The combined preparation of any one of clauses 76-77, or the method of clause 75 or 77, wherein the at least one additional active agent is a small molecule, peptide, an antibody, a corticosteroid, or a combination thereof.
Clause 79. The combined preparation or method of clause 78, wherein the at least one additional active agent is at least one of bortezomib, dexamethasone, and rituximab.
Clause 80. The combined preparation of any one of clauses 76-79, wherein the therapy is the treatment of cancer, autoimmune diseases, macular degeneration (MD) and related disorders, diseases and disorders associated with undesired angiogenesis, skin diseases, pulmonary disorders, asbestos-related disorders, parasitic diseases and disorders, immunodeficiency disorders, atherosclerosis and related conditions, hemoglobinopathy and related disorders, or TNFα related disorders.
| Number | Date | Country | Kind |
|---|---|---|---|
| PCT/PL2020/000099 | Dec 2020 | WO | international |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2021/087847 | 12/30/2021 | WO |
