Patent Application
20230065745
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 detrimental 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:
1) inhibition of cancer cell proliferation and induction of apoptosis,
2) disruption of trophic support from tumor stroma,
3) stimulation of immune cells, resulting in proliferation of T-cells, cytokine production and activation of NK (natural killer) cells (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).
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. There is thus a continuing need to provide novel cereblon-binding compounds which have pharmaceutically relevant properties.
Chemically-modified thalidomide-based derivatives, such as pomalidomide and lenalidomide, induce degradation of various neosubstrates, such as IKZF1, IKZF3, and/or CK1α. While degradation of IKZF1 and IKZF3 might be beneficial in the treatment of some cancer types (such as multiple myeloma), it may also contribute to dose-limiting toxicity of those compounds. Side effects resulting from lenalidomide's activity include neutropenia, thrombocytopenia, and hemorrhagic disorders (see: Sun X et al. PROTACs: great opportunities for academia and industry. Signal Transduct Target Ther. 2019 Dec. 24; 4:64 and Stahl M, Zeidan A M: Lenalidomide Use in Myelodysplastic Syndromes: Insights Into the Biologic Mechanisms and Clinical Applications. Cancer. 2017 May 15; 123(10):1703-1713). On the contrary, degradation of CK1α contributes to lenalidomide's therapeutic efficacy in myelodysplastic syndromes (see: Krönke J et al.: Lenalidomide induces ubiquitination and degradation of CK1α in del(5q) MDS. Nature. 2015 Jul. 9; 523(7559): 183-188. doi:10.1038/nature14610). Thus, new chemically-modified thalidomide-based derivatives capable of CK1α degradation and having more selective profile may be particularly useful in the treatment of cancer.
In accordance with a first aspect of the invention, there is provided a compound of Formula (Ia) or (Ib):
or a pharmaceutically acceptable salt or tautomer thereof,
In certain embodiments, the compound has the structure:
In other embodiments, the compound has the structure:
In some embodiments, T is C═O. In other embodiments, T is SO2.
In some embodiments, each R is independently hydrogen, halogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —NH2, —NHR″, —NR″2, —NR″C(O)R″, —NR″C(O)CH(OH)R″, —NR″C(O)OR″, —NR″SO2R″, —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″, or —S(O)2R″, —S(O)2OR″, —S(O)2NH2, —S(O)2NHR″, or —S(O)2NR″2.
In some embodiments, Z is S or NR2. In some embodiments, Z is NR2. In other embodiments, Z is S.
In some embodiments, L is hydrogen, alkyl, alkenyl, haloalkyl, haloalkenyl, —C(O)R″, —CH2C(O)OR″, —C(O)OR″, —C(O)NH2, —C(O)NHR″, —C(O)NR″2, —OR″, —NR″2, or —S(O)2R″. In some embodiments, L is hydrogen, alkyl, alkenyl, —CH2C(O)OR″, —OR″, —NR″2, or —S(O)2R″; optionally wherein L is hydrogen, alkyl, or alkenyl. In some embodiments, L is hydrogen, alkyl, —CH2C(O)OR″ or —OR″.
In some embodiments, L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —OR″, —NR″2, or —S(O)2R″. In other embodiments, 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, L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —OR″, —NR″2, or —S(O)2R″. In some embodiments, L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, or haloalkenyl. In other embodiments, L is —OR″, —NR″2, or —S(O)2R″ In some embodiments, L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, or benzyl. In some embodiments, L is hydrogen, alkyl, alkenyl, or aryl. In some embodiments, L is hydrogen, alkyl, or alkenyl. In some embodiments, L is hydrogen or alkyl. In some embodiments, L is hydrogen.
In some embodiments, R2 is hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —NR″2, —NR″C(O)R″, —N[C(O)R″]2, —NR″C(O)OR″, —C(O)R″, —C(O)OR″, —OR″, —OC(O)R″, —OC(O)OR″, —OC(O)NH2, —OC(O)NHR″, or —OC(O)NR″2. In some such embodiments, R2 is alkyl, benzyl, or —N[C(O)R″]2.
In some embodiments, the compound is of Formula (Ia) wherein one of Y1, Y2 and Y3 is N, and the remaining two of Y1, Y2 and Y3 are each CR. In some such embodiments, Y1 is N, and Y2 and Y3 are CR. In other such embodiments, Y2 is N; Y1 and Y3 are CR; and Z is S. In other such embodiments, Y3 is N; Y1 and Y2 are CR; and Z is S.
In some embodiments, the compound is of Formula (Ia), wherein one of Y1, Y2 and Y3 is CR; the remaining two of Y1, 112 and Y3 are each N; and Z is S. In some such embodiments, Y1 is CR; Y2 and Y3 are N. In other such embodiments, Y2 is CR; Y1 and Y3 are N. In other such embodiments, Y3 is CR, and Y1 and Y2 are N.
In some embodiments, the compound is of Formula (Ia) and Y1, Y2 and Y3 are each CR. In some such embodiments, Y1 is —C—NHC(O)R″, Y2 is CH, and Y3 is CH or CCl. In some such embodiments, L is hydrogen; Z is 5; R′ is H; T is C═O; Y1 is —C—NHC(O)R″; Y2 is CH; and Y3 is CH.
In some embodiments, the compound is of Formula (Ib), wherein one of Y1, Y2 and Y4 is N and the remaining two of Y1, 112 and Y4 are each CR, and wherein Z is S. In some such embodiments, Y1 is N, and Y2 and Y4 are CR. In other such embodiments, Y2 is N, and Y1 and Y4 are CR. In other such embodiments, Y4 is N, and Y1 and Y2 are CR.
In some embodiments, the compound is of Formula (Ib), wherein one of Y1, Y2 and Y4 is CR and the remaining two of Y1, Y2 and Y4 are each N, and wherein Z is S. In some such embodiments, Y1 is CR, and Y2 and Y4 are N. In other such embodiments, Y2 is CR, and Y1 and Y4 are N. In other such embodiments, Y4 is CR, and Y1 and Y2 are N.
In some embodiments, the compound is of Formula (Ib) and Y1, Y2 and Y4 are each CR. In some such embodiments, each R is independently hydrogen, halogen, alkyl, cycloalkyl, haloalkyl, heteroaryl, —OR″, —N[C(O)R″]2, —NR″C(O)R″, —NHC(O)OR″, —NHR″, —NH2, or —NHSO2R″CN. In some such embodiments, each R″ is independently alkyl, cycloalkyl, aryl or benzyl. In some such embodiments:
In some such embodiments each R″ is independently alkyl, cycloalkyl, aryl or benzyl. In some such embodiments, Y1 is CH; Y2 is CH or CCl; and Y4 is C—OR″ or C—NH2, optionally C—OMe or C—NH2.
In some embodiments of the compound of Formula (Ia) or Formula (Ib), each R is independently hydrogen, halogen, alkyl, cycloalkyl, haloalkyl, heteroaryl, —NR″C(O)R″, NR″C(O)OR″, —NR″C(O)CH(OH)R″, —NHR″, —NH2, —OR″, —CN, —C(O)NR″2, or —NR″SO2R″. In some such embodiments, each R is independently hydrogen, halogen, alkyl, cycloalkyl, haloalkyl, —OR″, —CN, —NHC(O)R″, —NHC(O)OR″, —NHR″, —NH2 or —NHSO2R″. In some embodiments, each R″ is independently alkyl, cycloalkyl, aryl or benzyl.
In some embodiments of the compound of Formula (Ia) or (Ib), X1 and X2 are O. In other embodiments, X1 is O and X2 is S. In other embodiments, X1 is S and X2 is 0. In other embodiments, X1 and X2 are S.
In some embodiments of the compound of Formula (Ia) or (Ib), n is 0. In other embodiments of the compound of Formula (Ia) or (Ib), n is 1 or 2. In some embodiments, n is 1. In other embodiments, n is 2.
In some embodiments, the compound is of Formula (Ia), wherein:
In some embodiments, the compound is of Formula (Ib), wherein:
In accordance with a second aspect of the invention, there is provided a compound of Formula (IIa) or (IIb):
or a pharmaceutically acceptable salt or tautomer thereof,
In some embodiments, the compound is of Formula (IIa).
In some embodiments, the compound is of Formula (IIb).
In some embodiments, the compound of Formula (IIa) or (IIb) has the structure:
In other embodiments, the compound of Formula (IIa) or (IIb) has the structure:
In some embodiments, when each is a double bond, Z is NR2, R2 is hydrogen, and Y3 and Y4 are CR, one of W1, W2, W3 and W4 is N, and the remaining three of W1, W2, W3 and W4 are each CR′, then at least one R′ is not hydrogen.
In some embodiments of the compound of Formula (IIa) or (IIb), Z is O. In other embodiments, Z is S. In other embodiments, Z is NR2.
In some embodiments of the compound of Formula (IIa) or (IIb), T is C═O. In other embodiments, T is SO2.
In some embodiments, L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, haloalkenyl, —C(O)R″, —C(O)OR″, —C(O)NH2, —C(O)NHR″, —C(O)NR″2, —NR″2, or —S(O)2R″; optionally wherein L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, —NR″2, or —S(O)2R″
In some embodiments of the compound of Formula (IIa) or (IIb), 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 (IIa) or (IIb), 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 (IIa) or (IIb), 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 (IIa) or (IIb), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, haloalkyl, or haloalkenyl. In other embodiments of the compound of Formula (IIa) or (IIb), L is —OR″, —NR″2, or —S(O)2R″ In some embodiments of the compound of Formula (IIa) or (IIb), L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, or benzyl. In some embodiments of the compound of Formula (IIa) or (IIb), L is hydrogen, alkyl, alkenyl, or aryl. In some embodiments of the compound of Formula (IIa) or (IIb), L is hydrogen, alkyl, or alkenyl. In some embodiments of the compound of Formula (IIa) or (IIb), L is hydrogen or alkyl. In some embodiments of the compound of Formula (IIa) or (IIb), L is hydrogen.
In some embodiments of the compound of Formula (IIa), Y3 is N. In other embodiments, Y3 is CR.
In some embodiments of the compound of Formula (IIb), Y4 is N. In other embodiments, Y4 is CR.
In some embodiments, indicates a single bond between W1 and W2, and W1 and W2 are each CR′2. In other embodiments,
indicates a double bond between W1 and W2, and W1 and W2 are each independently N or CR′. In either of the two aforementioned embodiments,
indicates a single bond between W3 and W4, and W3 and W4 are each CR′2. Alternatively,
indicates a double bond between W3 and W4, and W3 and W4 are each independently N or CR′.
In some embodiments of the compound of Formula (IIa) or (IIb), each is a double bond or each
is a single bond.
In some embodiments, each is a double bond. In some such 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, one of W1, W2 and W3 is N, and W4 is CR′. In some such embodiments W1 is N, and W2, W3 and W4 are each CR′. In other such embodiments, W2 is N, and W1, W3 and W4 are each CR′. In other such embodiments, W3 is N, and W1, W2 and W4 are each CR′. In other such embodiments, W4 is N, and W2 and W3 are each CR′. In other embodiments wherein each
is a double bond, two of W1, W2, W3 and W4 is 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 other embodiments wherein each is a double bond, one of W1, W2, W3 and W4 is CR′, and the remaining three of W1, W2, W3 and W4 are each N. In some such embodiments W1 is CR′, and W2, W3 and W4 are each N. In other such embodiments, W2 is CR′, and W1, W3 and W4 are each N. In other such embodiments, W3 is CR′, and W1, W2 and W4 are each N. In other such embodiments, W4 is CR′, and W1, W2 and W3 are each N.
In some embodiments, at least one R′ is not hydrogen.
In some embodiments of the compound of Formula (IIa) or (IIb), each is a single bond.
In some embodiments of the compound of Formula (IIa) or (IIb), each R is independently hydrogen, halogen or —NR″C(O)R″.
In some embodiments of the compound of Formula (IIa) or (IIb), each R′ is hydrogen.
In some embodiments of the compound of Formula (IIa) or (IIb), X1 and X2 are O. In other embodiments,
X1 is O and X2 is S. In other embodiments, X1 is S and X2 is 0. In other embodiments, X1 and X2 are S.
In some embodiments of the compound of Formula (IIa) or (IIb), n is 0. In other embodiments of the compound of Formula (IIa) or (IIb), n is 1 or 2. In some embodiments, n is 1. In other embodiments, n is 2.
In some embodiments, the compound is of Formula (IIa), wherein:
In some embodiments, the compound is of Formula (IIb), wherein:
In accordance with a third aspect, the present invention provides a compound of Formula (Ia), Formula (Ib), Formula (IIa) or Formula (IIb) selected from:
In accordance with a fourth 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, or a compound selected from
for use in medicine.
The invention also provides a compound or composition according to any of the above aspects of the present invention, or a compound selected from
for use in immune-oncology.
The invention also provides a compound or composition according to any of the above aspects of the present invention, or a compound selected from
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.
In some embodiments, the compound is a compound of any one of the first to third aspects of the present invention, or a compound selected from
In some embodiments, the compound is a compound of any one of the first to third aspects of the present invention.
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, or a compound selected from
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 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, an effective amount of a compound or composition according to any of the above aspects of the present invention is administered to the patient.
The present invention also provides a combined preparation of a compound of any one of the first to third aspects of the present invention, or a compound selected from
and at least one additional active agent, for simultaneous, separate or sequential use in therapy.
The present invention also provides a combined preparation of a compound of any one of the first to third 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 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. 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.
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 “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″, halogen, —NH2, —NHR″, —NR″2, —SO2R″, —C(O)R″, —CN, or —NO2. 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.
In some embodiments, all alkyl, cycloalkyl, heterocycolalkyl, alkenyl, alkynyl, aryl, heteroaryl, benzyl groups are unsubstituted.
As discussed above, the present invention provides compounds of Formulas (la), (Ib), (IIa) and (IIb):
wherein L, X1, X2, Y1, Y2, Y3, Y4, W1, W2, W3, W4, R1 and Z are as defined above.
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 in the following general procedure (carried out under Synthetic Conditions A or Synthetic Conditions B, as set out below:
Synthetic Conditions A
An appropriate acid (R′COOH in the above reaction scheme) (1.1 eq), DMAP (0.04 eq), and EDC (1.2 eq) were added to a solution of 3-aminopiperidine-2,6-dione (1 eq) and N-hydroxybenzotriazole (1.2 eq) in DMF (0.5 M). The reaction mixture was stirred overnight at room temperature (20-25° C.). Water (2×DMF volume) was added and the obtained solution was extracted with dichloromethane (3×DMF volume). The combined organic layers were washed with water, dried over Na2SO4, and concentrated under reduced pressure. The crude product was purified by preparative HPLC or by column chromatography.
Synthetic Conditions B
An appropriate acid RxCOOH in the above reaction scheme) (1 eq) and EDC (1.2 eq) were added to a solution of 3-aminopiperidine-2,6-dione (hydrochloride salt, 1.1 eq), triethylamine (1.2 eq) and N-hydroxybenzotriazole (1.2 eq) in DMA (0.5 M). The reaction mixture was stirred overnight at rt. Water (2×DMA volume) was added and obtained mixture was extracted with dichloromethane (3×DMA volume). The combined organic layers were washed with water, dried over Na2SO4, and concentrated under reduced pressure. The crude product was isolated by preparative HPLC or by column chromatography.
NCS (1.1 eq) was added to a solution of an appropriate starting material (1 eq) in DMF (0.5 M) and the reaction mixture was stirred for 2 h at room temperature (20-25° C.). The reaction mixture was poured into water (2×DMF volume) and occurred precipitate was filtered. The solids were washed with water and dried in vacuum to give the acid, ROOH.
LiOH (1.1 eq) was added to a solution of an appropriate ester (1 eq) in THF:water mixture (3:1 or 5:1, 85 mM) and the resulting mixture was stirred overnight at room temperature (20-25° C.). The mixture was concentrated under reduced pressure, diluted with water, and acidified with concentrated HCl to pH=2-3. The precipitate was filtered, washed with water, and dried in vacuum to give the target carboxylic acid.
A mixture of an appropriate amine (1 eq.), Ac2O (3 eq.), and DMAP (0.2 eq.) in dioxane (0.2 M) was heated to 80° C. for 2 h. Upon completion, the mixture was cooled down to room temperature (20-25° C.) and concentrated under reduced pressure. The residue was diluted with water (1× dioxane volume) and extracted with EtOAc (3× dioxane volume). The organic layers were washed with water, brine, dried over Na2SO4, and evaporated to dryness to afford an acylated product typically used without further purification.
Some examples of compounds of the present invention are shown below:
As also discussed in the Examples section, the present inventors have found that compounds of the present invention exhibits similar cereblon binding capabilities 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.
The compounds of the present invention are advantageous in terms of their synthetic feasibility. The synthesis of the compounds can be summarized in the following general procedure (carried out under Synthetic Conditions A or Synthetic Conditions B, as set out below:
Synthetic Conditions A
An appropriate acid (RCOOH in the above reaction scheme) (1.1 eq), DMAP (0.04 eq), and EDC (1.2 eq) were added to a solution of 3-aminopiperidine-2,6-dione (1 eq) and N-hydroxybenzotriazole (1.2 eq) in DMF (0.5 M). The reaction mixture was stirred overnight at room temperature (20-25° C.). Water (2×DMF volume) was added and the obtained solution was extracted with dichloromethane (3×DMF volume). The combined organic layers were washed with water, dried over Na2SO4, and concentrated under reduced pressure. The crude product was purified by preparative HPLC or by column chromatography.
Synthetic Conditions B
An appropriate acid RCOOH in the above reaction scheme) (1 eq) and EDC (1.2 eq) were added to a solution of 3-aminopiperidine-2,6-dione (hydrochloride salt, 1.1 eq), triethylamine (1.2 eq) and N-hydroxybenzotriazole (1.2 eq) in DMA (0.5 M). The reaction mixture was stirred overnight at rt. Water (2×DMA volume) was added and obtained mixture was extracted with dichloromethane (3×DMA volume). The combined organic layers were washed with water, dried over Na2SO4, and concentrated under reduced pressure. The crude product was isolated by preparative HPLC or by column chromatography.
Synthetic Conditions C
To a solution of appropriate acid (RCOOH in the above reaction scheme) (1 eq) and HATU (1.5 eq) in dry DMF were added 3-aminopiperidine-2,6-dione (hydrochloride salt, 1.2 eq) and DIPEA (3 eq). The reaction mixture was stirred overnight at rt. The crude product was purified by preparative HPLC or/and by preparative TLC.
Synthetic Conditions D
To a solution of appropriate acid (RCOOH in the above reaction scheme) (1 eq) 3-aminopiperidine-2,6-dione (hydrochloride salt, 1.2 eq) and DMAP (0.1 eq.) in an inert atmosphere in dry DMF were added DIPEA (2.2 eq.) and HATU (1.5 eq) in dry DMF. The reaction mixture was stirred overnight at rt. The crude product was purified by preparative HPLC or/and by preparative TLC.
N-chlorosuccinimide (1.1 eq) was added to a solution of an appropriate starting material (1 eq) in DMF (0.5 M) and the reaction mixture was stirred for 2 h at room temperature (20-25° C.). The reaction mixture was poured into water (2×DMF volume) and occurred precipitate was filtered. The solids were washed with water and dried in vacuum to give the acid, ROOH.
LiOH (1.1 eq) was added to a solution of an appropriate ester (1 eq) in THF:water mixture (3:1 or 5:1, 85 mM) and the resulting mixture was stirred overnight at room temperature (20-25° C.). The mixture was concentrated under reduced pressure, diluted with water, and acidified with concentrated HCl to pH=2-3. The precipitate was filtered, washed with water, and dried in vacuum to give the target carboxylic acid.
A mixture of an appropriate amine (1 eq.), Ac2O (3 eq.), and DMAP (0.2 eq.) in dioxane (0.2 M) was heated to 80° C. for 2 h. Upon completion, the mixture was cooled down to room temperature (20-25° C.) and concentrated under reduced pressure. The residue was diluted with water (1× dioxane volume) and extracted with EtOAc (3× dioxane volume). The organic layers were washed with water, brine, dried over Na2SO4, and evaporated to dryness to afford an acylated product typically used without further purification.
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (52% yield) using 2-((tert-butoxycarbonyl)amino)thiophene-3-carboxylic acid (43 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 11.09 (s, 1H), 10.88 (s, 1H), 8.63 (d, J=8.2 Hz, 1H), 7.38 (d, J=5.9 Hz, 1H), 6.97 (dd, J=5.8, 0.8 Hz, 1H), 4.68 (ddd, J=13.0, 8.2, 5.3 Hz, 1H), 2.78 (ddd, J=17.3, 13.5, 5.5 Hz, 1H), 2.59-2.54 (m, 1H), 2.15 (qd, J=13.0, 4.5 Hz, 1H), 1.96 (ddt, J=10.2, 5.3, 2.7 Hz, 1H), 1.48 (s, 9H).
LCMS (m/z [M+H]−): 352.0
Step A: 3-Aminopiperidine-2,6-dione (3.3 g, 25.8 mmol) and triethylamine (2.45 g, 24.2 mmol) were added to a solution of 1-methyl-1H,2H,4H-thieno[2,3-d][1,3]oxazine-2,4-dione (3.7 g, 20.2 mmol) in ethanol (20 mL). The reaction mixture was refluxed for 16 h and filtered. The precipitate was washed with water to give N-(2,6-dioxopiperidin-3-yl)-2-(methylamino)thiophene-3-carboxamide (19% yield).
Step B: Acetic anhydride (0.265 g, 2.60 mmol) and DMAP (0.026 g, 0.213 mmol) were added to a solution of N-(2,6-dioxopiperidin-3-yl)-2-(methylamino)thiophene-3-carboxamide (0.579 g, 2.17 mmol) and triethylamine (0.263 g, 2.60 mmol) in dioxane (10 mL). The reaction mixture was stirred at 60° C. for 16 h, washed with water and extracted with EtOAc (3×10 mL), dried over Na2SO4, concentrated under reduced pressure and purified by HPLC to give N-(2,6-dioxopiperidin-3-yl)-2-(N-methylacetamido)thiophene-3-carboxamide (11% yield).
1H NMR (500 MHz, DMSO) δ 10.86 (s, 1H), 8.53 (d, J=8.0 Hz, 1H), 7.56 (d, J=5.7 Hz, 1H), 7.34 (d, J=5.7 Hz, 1H), 4.72-4.62 (m, 1H), 3.09 (s, 3H), 2.83-2.71 (m, 1H), 2.58-2.53 (m, 1H), 2.19-2.02 (m, 1H), 1.99-1.90 (m, 1H), 1.81 (s, 3H).
LCMS (m/z [M+H]+): 310.2
Step A: Methyl 5-chloro-2-pentanamidothiophene-3-carboxylate was synthesized using Example Method 1, above (65% yield), using methyl 2-pentanamidothiophene-3-carboxylate as a starting material.
Step B: 5-chloro-2-pentanamidothiophene-3-carboxylic acid was synthesized using Example
Method 2, above (69% yield), using methyl 5-chloro-2-pentanamidothiophene-3-carboxylate as a starting material.
Step C: 5-chloro-N-(2,6-dioxopiperidin-3-yl)-2-pentanamidothiophene-3-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions A, above (40% yield), and 5-chloro-2-pentanamidothiophene-3-carboxylic acid as a starting material.
1H NMR (400 MHz, DMSO) δ 11.94 (s, 1H), 10.92 (s, 1H), 8.72-8.59 (m, 1H), 7.52 (s, 1H), 4.84-4.71 (m, 1H), 2.96-2.59 (m, 4H), 2.61-2.53 (m, 1H), 2.23-2.06 (m, 1H), 2.04-1.91 (m, 2H), 1.68-1.51 (m, 2H), 1.42-1.26 (m, 2H), 1.02-0.81 (m, 3H).
LCMS (m/z [M+H]+): 372.2
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above, (32% yield) with 2-acetamidothiophene-3-carboxylic acid as a starting material.
1H NMR (400 MHz, DMSO) δ 11.84 (s, 1H), 10.90 (s, 1H), 8.67 (d, J=8.1 Hz, 1H), 7.42 (d, J=5.8 Hz, 1H), 7.00 (d, J=5.7 Hz, 1H), 4.79-4.67 (m, 1H), 2.85-2.72 (m, 1H), 2.61-2.54 (m, 1H), 2.20 (s, 3H), 2.17-2.11 (m, 1H), 1.96-1.84 (m, 1H).
LCMS (m/z [M+H]+): 296.06
Triethylamine (0.236 g, 2.328 mmol), N-hydroxybenzotriazole (0.314, 2.3 mmol), 2-acetamidothiophene-3-carboxylic acid (0.359 g, 1.94 mmol) and EDC (0.361 g, 2.328 mmol) were added sequentially to a solution of 3-(methylamino)piperidine-2,6-dione (0.381 g, 2.134 mmol, hydrochloride salt) in DMA (30 mL) and the reaction mixture was stirred overnight at room temperature. Water (10 mL) was added and obtained solution was extracted with DCM, dried over Na2SO4 and concentrated under reduced pressure. The product was purified by HPLC to give N-(2,6-dioxopiperidin-3-yl)-2-acetamido-N-methylthiophene-3-carboxamide (25% yield).
1H NMR (400 MHz, DMSO) δ 10.99 (s, 1H), 10.54 (s, 1H), 7.13-7.02 (m, 1H), 7.01-6.89 (m, 1H), 5.13-4.87 (m, 1H), 2.87 (s, 3H), 2.83-2.71 (m, 1H), 2.61-2.52 (m, 2H), 2.15 (s, 3H), 2.07-1.95 (m, 1H).
LCMS (m/z [M+H]+): 310.0
Step A: Methyl 5-chloro-2-cyclopropaneamidothiophene-3-carboxylate was synthesized using Example Method 1, above (80% yield) using methyl 2-cyclopropaneamidothiophene-3-carboxylate as a starting material.
Step B: 5-chloro-2-cyclopropaneamidothiophene-3-carboxylic acid was synthesized using Example Method 2, above (86% yield) using methyl 5-chloro-2-cyclopropaneamidothiophene-3-carboxylate as a starting material.
Step C: 5-chloro-2-cyclopropaneamido-N-(2,6-dioxopiperidin-3-yl)thiophene-3-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above (30% yield), using 5-chloro-2-cyclopropaneamidothiophene-3-carboxylic acid as a starting material.
1H NMR (500 MHz, DMSO) δ 12.11 (s, 1H), 10.88 (s, 1H), 8.69-8.60 (m, 1H), 7.50 (s, 1H), 4.79-4.69 (m, 1H), 2.84-2.72 (m, 1H), 2.61-2.53 (m, 1H), 2.21-2.08 (m, 1H), 2.04-1.92 (m, 2H), 0.98-0.84 (m, 4H).
LCMS (m/z [M+H]+): 356.2
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above (15% yield), and N-(2,6-dioxopiperidin-3-yl)-3-acetamidothiophene-2-carboxamide as a starting material.
1H NMR (400 MHz, DMSO) δ 10.89 (s, 2H), 8.55 (d, J=8.1 Hz, 1H), 7.94 (d, J=5.3 Hz, 1H), 7.75 (d, J=5.4 Hz, 1H), 4.76-4.66 (m, 1H), 2.85-2.69 (m, 1H), 2.59-2.52 (m, 1H), 2.25-2.12 (m, 1H), 2.09 (s, 3H), 2.01-1.89 (m, 1H).
LCMS (m/z [M+H]+): 295.8
Step A: H2SO4 (1 mL) was added dropwise to a stirred suspension of methyl 2-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino)-4-oxo-4,5-dihydrothiophene-3-carboxylate (9.65 g, 24.4 mmol) in MeOH (200 mL). The reaction mixture was refluxed for 16 h, cooled to RT and filtered to give 2-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino)-4-methoxythiophene-3-carboxylate (63% yield).
Step B: Morpholine (13.5 g, 155 mmol) was added to a solution of methyl 2-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino)-4-methoxythiophene-3-carboxylate (6.3 g, 15.4 mmol) in dichloromethane (100 mL) and the reaction mixture was stirred overnight at room temperature, concentrated under reduced pressure, diluted with MTBE, filtered, and rinsed with small amount of MTBE. The filtrate was evaporated in vacuo to give crude methyl 2-amino-4-methoxythiophene-3-carboxylate, which was used in the next step without further purification.
Step C: methyl 2-acetamido-4-methoxythiophene-3-carboxylate was obtained in 73% yield using Example Method 3, above, with methyl 2-amino-4-methoxythiophene-3-carboxylate as a starting material.
Step D: 2-acetamido-4-methoxythiophene-3-carboxylic acid was obtained in 20% yield using Example Method 2, above, with methyl 2-acetamido-4-methoxythiophene-3-carboxylate as a starting material.
Step E: N-(2,6-dioxopiperidin-3-yl)-2-acetamido-4-methoxythiophene-3-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above (47% yield), and 2-acetamido-4-methoxythiophene-3-carboxylic acid as a starting material.
1H NMR (500 MHz, DMSO) δ 12.05 (s, 1H), 10.92 (s, 1H), 8.30 (d, J=7.1 Hz, 1H), 6.14 (s, 1H), 4.76-4.66 (m, 1H), 3.83 (s, 3H), 2.82-2.70 (m, 1H), 2.58-2.52 (m, 1H), 2.19 (s, 3H), 2.16-2.06 (m, 2H)
LCMS (m/z [M+H]+): 326.2
Step A: Ethyl 2-acetamidothiophene-3-carboxylate (11 g, 51.6 mmol) was dissolved in AcOH (110 mL) and solution of bromine (3.2 mL, 61.9 mmol) in AcOH (55 mL) was added dropwise over 15 min at RT. The reaction mixture was stirred at RT for 18 h, concentrated under reduced pressure and diluted water. The precipitate was filtered, washed with water and dried to give ethyl 5-bromo-2-acetamidothiophene-3-carboxylate (93% yield).
Step B: Zn(CN)2 (8.45 g, 72 mmol) and Pd(dppf)Cl2DCM (3.92 g, 4.8 mmol) were added to a solution of ethyl 5-bromo-2-acetamidothiophene-3-carboxylate (14 g, 48 mmol) in DMF (120 mL). Argon was bubbled through the reaction mixture for 10 min, then the reaction mixture was stirred at 150° C. for 16 h, cooled to RT, filtered and washed with EtOAc. The organic layer was dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give 5-cyano-2-acetamidothiophene-3-carboxylate (83% yield).
Step C: Ethyl 5-cyano-2-acetamidothiophene-3-carboxylate (9.45 g, 39.7 mmol) was dissolved in EtOH:THF solution (120 mL:360 mL), the solution was cooled to +5° C. and lithium hydroxide monohydrate (11.7 g, 278 mmol) in H2O (120 mL) was added dropwise over 20 min. The reaction mixture was stirred at RT for 18 h, concentrated under reduced pressure and acidified with 15% citric acid. The product was extracted with EtOAc, dried over Na2SO4 and evaporated under reduced pressure to give 5-cyano-2-acetamidothiophene-3-carboxylic acid (57% yield).
Step D: 5-cyano-N-(2,6-dioxopiperidin-3-yl)-2-acetamidothiophene-3-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions A, above (35% yield), and 5-cyano-2-acetamidothiophene-3-carboxylic acid as a starting material.
1H NMR (400 MHz, DMSO) δ 12.08 (s, 1H), 10.94 (s, 1H), 8.89 (d, J=7.9 Hz, 1H), 8.27 (s, 1H), 4.82-4.65 (m, 1H), 2.87-2.72 (m, 1H), 2.62-2.53 (m, 1H), 2.30 (s, 3H), 2.24-2.08 (m, 1H), 2.06-1.93 (m, 1H)
LCMS (m/z [M+H]+): 321.0
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above (23% yield), and 4-chloro-N-(2,6-dioxopiperidin-3-yl)-2-acetamidothiophene-3-carboxamide as a starting material.
1H NMR: (400 MHz, DMSO-d6) δ 11.21 (s, 1H), 11.05 (s, 1H), 8.30 (d, J=7.7 Hz, 1H), 6.57 (s, 1H), 4.94-4.79 (m, 1H), 2.88-2.74 (m, 1H), 2.63-2.52 (m, 1H), 2.16 (5, 3H), 2.13-2.04 (m, 2H), 2.03-1.95 (m, 2H), 0.98-0.83 (m, 2H), 0.72-0.64 (m, 1H), 0.64-0.55 (m, 1H)
LCMS (m/z [M+H]′): 336.3
Step A: methyl 4,5-dichloro-2-acetamidothiophene-3-carboxylate was prepared in 90% yield using Example Method 1, above, with methyl 4-chloro-2-acetamidothiophene-3-carboxylate as a starting material.
Step B: 4,5-Dichloro-2-acetamidothiophene-3-carboxylic acid was prepared in 70% yield using Example Method 2, above, with methyl 4,5-chloro-2-acetamidothiophene-3-carboxylate as a starting material.
Step C: 4,5-dichloro-N-(2,6-dioxopiperidin-3-yl)-2-acetamidothiophene-3-carboxamide was synthesized in 40% yield using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions A, above, and 4,5-dichloro-2-acetamidothiophene-3-carboxylic acid as a starting material.
1H NMR: (500 MHz, DMSO) d 11.20 (s, 1H), 11.09 (s, 1H), 8.74-8.67 (m, 1H), 4.93-4.80 (m, 1H), 2.88-2.75 (m, 1H), 2.62-2.52 (m, 1H), 2.19 (s, 3H), 2.14-2.02 (m, 2H).
LCMS (m/z [M−H]−): 362.1
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B above (22% yield), with 5-chloro-2-acetamidothiophene-3-carboxylic acid as a starting material.
1H NMR (500 MHz, DMSO) δ 11.84 (s, 1H), 10.91 (s, 1H), 8.67 (d, J=8.0 Hz 1H), 7.50 (s, 1H), 4.77-4.65 (m, 1H), 2.84-2.73 (m, 1H), 2.61-2.54 (m, 1H), 2.22 (s, 3H), 2.19-2.07 (m, 1H), 2.02-1.91 (m, 1H)
LCMS (m/z [M+H]+): 329.8
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above (26% yield), with 2-(methylsulfonamido)thiophene-3-carboxylic acid as a starting material.
1H NMR (400 MHz, DMSO) δ 11.12 (s, 1H), 10.92 (s, 1H), 8.80-8.65 (m, 1H), 7.46-7.36 (m, 1H), 7.17-7.06 (m, 1H), 4.82-4.69 (m, 1H), 3.17 (s, 3H), 2.85-2.72 (m, 1H), 2.60-2.53 (m, 1H), 2.20-2.04 (m, 1H), 2.02-1.92 (m, 1H)
LCMS (m/z [M+H]+): 332.2
Step A: 4-tert-butyl 2-ethyl 5-aminothiophene-2,4-dicarboxylate (3.71 g, 13.7 mmol) was added to 20% solution of methylamine in methanol (20 mL) and the reaction mixture was stirred for 5 days at 70° C., concentrated under reduced pressure and triturated with isopropyl alcohol:hexane (1:1). The precipitate was filtered to give tert-butyl 2-amino-5-(methylcarbamoyl)thiophene-3-carboxylate (93% yield).
Step B: Triethylamine (3.3 g, 32.6 mmol), DMAP (0.13 g, 1.06 mmol) and acetic acid (1.67 g, 27.8 mmol) were added to a solution of tert-butyl 2-amino-5-(methylcarbamoyl)thiophene-3-carboxylate (2.8 g, 10.9 mmol) in dry MeCN (30 mL). The reaction mixture was stirred overnight at 50° C., cooled to room temperature, diluted with water, extracted with DCM, dried over Na2SO4 and concentrated under reduced pressure to give tert-butyl 2-acetamido-5-(methylcarbamoyl)thiophene-3-carboxylate (95% yield).
Step C: 10% HCl in dioxane (20 mL) was added to a solution of tert-butyl 2-acetamido-5-(methylcarbamoyl)thiophene-3-carboxylate (3.1 g, 10.4 mmol) in DCM (20 mL) and the reaction mixture was stirred for 3 days at RT. The precipitate was filtered, washed with DCM and dried to give 2-acetamido-5-(methylcarbamoyl)thiophene-3-carboxylic acid (60% yield).
Step D: N4-(2,6-dioxopiperidin-3-yl)-5-acetamido-N2-methylthiophene-2,4-dicarboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above (44% yield), and 2-acetamido-5-(methylcarbamoyl)thiophene-3-carboxylic acid as a starting material.
1H NMR (400 MHz, DMSO) δ 11.84 (s, 1H), 10.92 (s, 1H), 8.78 (d, J=8.1 Hz, 1H), 8.35-8.25 (m, 1H), 7.96 (s, 1H), 4.81-4.68 (m, 1H), 2.85-2.74 (m, 1H), 2.73 (d, J=4.4 Hz, 3H), 2.62-2.52 (m, 1H), 2.24 (s, 3H), 2.20-2.07 (m, 1H), 2.04-1.93 (m, 1H)
LCMS (m/z [M+H]+): 352.9
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (24% yield) using 2-oxo-2,3-dihydrothiazole-4-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 11.62 (s, 1H), 10.89 (s, 1H), 8.69 (d, J=8.3 Hz, 1H), 7.17 (s, 1H), 4.68 (ddd, =12.5, 8.3, 5.4 Hz, 1H), 2.78 (ddd, =17.3, 13.2, 5.7 Hz, 1H), 2.60-2.52 (m, 1H), 2.11-2.00 (m, 1H), 1.96 (dtd, J=12.7, 5.5, 2.8 Hz, 1H).
LCMS (m/z [M+H]+): 256.2
Step A: N-chlorosuccinimide (0.884 g, 6.62 mmol) was added to a solution of 1-methyl-1H,2H,4H-thieno[2,3-d][1,3]oxazine-2,4-dione (1 g, 5.46 mmol) in mixture of toluene (4 mL) and acetic acid (4 mL). The reaction mixture was stirred at 70° C. for 2 h, concentrated under reduced pressure, diluted with water and filtered. The solids were washed with water and dried 6-chloro-1-methyl-1H,2H,4H-thieno[2,3-d][1,3]oxazine-2,4-dione (72% yield).
Step B: 3-Aminopiperidine-2,6-dione hydrochloride (0.655 g, 3.98 mmol) and triethylamine (0.483 g, 4.77 mmol) were added to a solution of 6-chloro-1-methyl-1H,2H,4H-thieno[2,3-d][1,3]oxazine-2,4-dione (0.865 g, 3.97 mmol) in ethanol (20 mL) and the reaction mixture was refluxed for 18 h, concentrated under reduced pressure and diluted with water. The precipitate was filtered, washed with water and isopropyl alcohol, and dried to give 5-chloro-N-(2,6-dioxopiperidin-3-yl)-2-(methylamino)thiophene-3-carboxamide (44% yield).
1H NMR (400 MHz, DMSO) δ 10.79 (s, 1H), 8.18-8.04 (m, 1H), 8.03-7.91 (m, 1H), 7.26 (s, 1H), 4.68-4.52 (m, 1H), 2.85 (s, 3H), 2.79-2.67 (m, 1H), 2.60-2.53 (m, 1H), 2.16-2.01 (m, 1H), 1.99-1.85 (m, 1H).
LCMS (m/z [M+H]+): 302.2
Step A: Oxalyl chloride (9.24 g, 72.8 mmol) and a drop of DMF were added to a solution of (2S)-2-[(tert-butyldimethylsilyl)oxy]propanoic acid (11.9 g, 58.2 mmol) in dry DCM (150 mL). The resulting mixture was stirred at RT for 2 h, concentrated under reduced pressure, dissolved in DCM (50 mL) and added dropwise to a cooled solution of methyl 2-aminothiophene-3-carboxylate (4.58 g, 29.1 mmol) and DIPEA (11.3 g, 87.4 mmol) in DCM (150 mL). The reaction mixture was stirred at RT for 2 h, washed with water, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give methyl 2-[(2S)-2-[(tert-butyldimethylsilyl)oxy]propanamido]thiophene-3-carboxylate (24% yield).
Step B: N-chlorosuccinimide (1.03 g, 7.71 mmol) was added to a solution of methyl 2-[(2.5)-2-[(tert-butyldimethylsilyl)oxy]propanamido]thiophene-3-carboxylate (2.4 g, 6.99 mmol) in DMF (30 mL) and the mixture was stirred at RT for 18 h. The reaction mixture was poured into water and extracted with EtOAc, dried over Na2SO4 and concentrated under reduced pressure to give methyl 2-[(2S)-2-[(tert-butyldimethylsilyl)oxy]propanamido]-5-chlorothiophene-3-carboxylate (91% yield).
Step C: 10% aqueous solution of LiOH (6 mL) was added to a solution of crude methyl 2-[(2S)-2-[(tert-butyldimethylsilyl)oxy]propanamido]-5-chlorothiophene-3-carboxylate (2.42 g, 6.40 mmol) in THF (12 mL) and the mixture was stirred at RT for 3 days. The reaction mixture was concentrated under reduced pressure, diluted with water and acidified with 10% HCl. The product was extracted into DCM, dried over Na2SO4, concentrated under reduced pressure and crystallized to give (S)-5-chloro-2-(2-hydroxypropanamido)thiophene-3-carboxylic acid (25% yield).
Step D: 3-Aminopiperidine-2,6-dione (0.421 g, 2.56 mmol, hydrochloride salt), 3H-[1,2,3]triazolo[4,5-b]pyridin-3-ol (0.191 g, 1.41 mmol), triethylamine (0.330 g, 3.27 mmol), and EDC (0.397 g, 2.56 mmol) were added sequentially to a solution of (S)-5-chloro-2-(2-hydroxypropanamido)thiophene-3-carboxylic acid (0.319 g, 1.28 mmol) in DMA (2.5 mL). The reaction mixture stirred at RT for 18 h, concentrated under reduced pressure and purified by HPLC to give 5-chloro-N-(2,6-dioxopiperidin-3-yl)-2-[(2S)-2-hydroxypropanamido]thiophene-3-carboxamide (29% yield).
1H NMR (400 MHz, DMSO) δ 12.59 (s, 1H), 10.91 (s, 1H), 8.65 (d, J=8.3 Hz, 1H), 7.54 (s, 1H), 6.25 (d, J=4.8 Hz, 1H), 4.82-4.71 (m, 1H), 4.34-4.23 (m, 1H), 2.85-2.71 (m, 1H), 2.61-2.53 (m, 1H), 2.20-2.04 (m, 1H), 2.01-1.90 (m, 1H), 1.32 (d, 3H)
LCMS (m/z [M−1-1]+): 359.9
Step A: 5-chloro-3-acetamidothiophene-2-carboxylic acid was obtained in 72% yield using Example Method 2, above, with methyl 5-chloro-3-acetamidothiophene-2-carboxylate as a starting material.
Step B: 5-chloro-N-(2,6-dioxopiperidin-3-yl)-3-acetamidothiophene-2-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions A, above (40% yield), and 5-chloro-3-acetamidothiophene-2-carboxylic acid as a starting material.
1H NMR (400 MHz, DMSO) δ 10.91 (s, 1H), 10.87 (s, 1H), 8.61 (d, J=8.2 Hz, 1H), 7.90 (s, 1H), 4.75-4.64 (m, 1H), 2.85-2.70 (m, 1H), 2.60-2.52 (m, 2H), 2.11 (s, 3H), 2.00-1.91 (m, 1H)
LCMS (m/z [M+H]+): 330.2
Step A: SO2Cl2 (0.207 g, 1.53 mmol) was added to a solution of methyl 5-cyclopropyl-2-acetamidothiophene-3-carboxylate (0.306 g, 1.28 mmol) in CHCl3 (15 mL). The reaction mixture was refluxed for 2 h, concentrated under reduced pressure and diluted with water. The product was extracted with EtOAc, dried over Na2SO4 and concentrated under reduced pressure to give methyl 4-chloro-5-cyclopropyl-2-acetamidothiophene-3-carboxylate (81% yield).
Step B: 4-chloro-5-cyclopropyl-2-acetamidothiophene-3-carboxylic acid was obtained in 78% yield using Example Method 2, above, with methyl 4-chloro-5-cyclopropyl-2-acetamidothiophene-3-carboxylate as a starting material.
Step C: HATU (0.370 g, 0.973 mmol) was added to the solution of 4-chloro-5-cyclopropyl-2-acetamidothiophene-3-carboxylic acid (0.211 g, 0.812 mmol), 3-aminopiperidine-2,6-dione (0.134 g, 1.05 mmol) and N-methylmorpholine (0.205 g, 2.03 mmol) in DMF (5 mL) at 0° C. The reaction mixture was stirred overnight at room temperature, diluted with water, extracted with AcOEt, dried over Na2SO4, concentrated under reduced pressure and purified by HPLC to give 4-chloro-5-cyclopropyl-N-(2,6-dioxopiperidin-3-yl)-2-acetamidothiophene-3-carboxamide (41% yield).
1H NMR (400 MHz, DMSO) δ 11.07 (s, 1H), 11.04 (s, 1H), 8.54 (d, J=8.2 Hz, 1H), 4.90-4.78 (m, 1H), 2.89-2.72 (m, 1H), 2.65-2.52 (m, 2H), 2.16 (s, 3H), 2.12-1.98 (m, 2H), 1.08-0.98 (m, 2H), 0.71-0.58 (m, 2H).
LCMS (m/z [M+H]+): 369.8
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above (17% yield), using 2-methoxythiophene-3-carboxylic acid as a starting material.
1H NMR (400 MHz, DMSO) δ 10.86 (s, 1H), 7.81 (d, J=7.5 Hz, 1H), 7.10 (d, J=5.9 Hz, 1H), 6.82 (d, J=5.9 Hz, 1H), 4.74-4.63 (m, 1H), 4.09 (s, 3H), 2.83-2.69 (m, 1H), 2.57-2.51 (m, 1H), 2.18-1.99 (m, 2H)
LCMS (m/z [M+H]+): 269.0
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions D, above, (54% yield) using 2-methoxythiophene-3-carboxylic acid (20 mg) and (S)-3-aminopiperidine-2,6-dione as a starting material.
1H NMR (500 MHz, DMSO) δ 10.86 (s, 1H), 7.81 (d, I=7.5 Hz, 1H), 7.11 (d, J=5.9 Hz, 1H), 6.83 (d, J=5.9 Hz, 1H), 4.76-4.61 (m, 1H), 4.05 (s, 3H), 2.77 (ddd, J=17.4, 13.3, 6.0 Hz, 1H), 2.48-2.44 (m, 1H), 2.17-2.01 (m, 2H).
LCMS (m/z [M+H]+): 268.9
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions D, above, (49.5% yield) using 2-methoxythiophene-3-carboxylic acid (20 mg) and (R)-3-aminopiperidine-2,6-dione as a starting material.
1H NMR (500 MHz, DMSO) δ 10.86 (s, 1H), 7.81 (d, J=7.5 Hz, 1H), 7.11 (d, J=5.9 Hz, 1H), 6.83 (d, J=5.9 Hz, 1H), 4.78-4.52 (m, 1H), 4.05 (s, 3H), 2.77 (ddd, J=17.4, 13.3, 6.0 Hz, 1H), 2.48-2.45 (m, 1H), 2.17-1.99 (m, 2H).
LCMS (m/z [M+H]+): 268.9
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (61% yield) using 2-methoxythiophene-3-carboxylic acid (20 mg) and 3-amino-1-methylpiperidine-2,6-dione trifluoroacetic acid salt (1.2 eq.) as a starting material.
1H NMR (500 MHz, DMSO) δ 7.86 (d, J=7.5 Hz, 1H), 7.11 (d, J=5.9 Hz, 1H), 6.82 (d, J=5.9 Hz, 1H), 4.76 (ddd, J=12.2, 7.4, 5.7 Hz, 1H), 4.05 (s, 3H), 3.01 (s, 3H), 2.85 (ddd, J=17.4, 13.1, 6.1 Hz, 1H), 2.67 (ddd, J=17.3, 4.5, 2.9 Hz, 1H), 2.17-2.01 (m, 2H).
LCMS (m/z [M+H]+): 283.1
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (44% yield) using 2-methoxythiophene-3-carboxylic acid (25.5 mg, 1.1 eq.) and methyl 2-(3-amino-2,6-dioxopiperidin-1-yl)acetate (trifluoroacetic acid salt, 1.0 eq.) as a starting materials.
1H NMR (500 MHz, DMSO) δ 7.88 (d, J=7.6 Hz, 1H), 7.11 (d, J=6.0 Hz, 1H), 6.83 (d, J=5.9 Hz, 1H), 4.84 (ddd, J=12.8, 7.6, 5.4 Hz, 1H), 4.50-4.34 (m, 2H), 4.05 (s, 3H), 3.66 (s, 3H), 3.03-2.91 (m, 1H), 2.76 (ddd, J=17.6, 4.3, 2.5 Hz, 1H), 2.24-2.13 (m, 1H), 2.13-2.05 (m, 1H).
LCMS (m/z [M+H]+): 341.2
To the cooled to 0° C. mixture of 3-aminopiperidine-2,6-dione hydrochloride (1.5 eq) and triethylamine (5 eq) in DCM (1.5 mL) was added 2-methoxythiophene-3-sulfonyl chloride (20 mg). Reaction was stirred for 18 h at RT, concentrated under reduced pressure and purified by flash column chromatography to give N-(2,6-dioxopiperidin-3-yl)-2-methoxythiophene-3-sulfonamide (66% yield).
1H NMR (500 MHz, DMSO) δ 10.77 (s, 1H), 7.79 (d, J=8.0 Hz, 1H), 6.98 (d, J=6.0 Hz, 1H), 6.86 (d, J=6.0 Hz, 1H), 4.15 (dtd, J=11.9, 9.3, 8.7, 5.8 Hz, 1H), 3.99 (s, 3H), 2.68 (td, J=12.1, 6.1 Hz, 1H), 2.48-2.44 (m, 1H), 1.92-1.78 (m, 2H).
LCMS (m/z [M+H]+): 305.1
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions D, above, (78% yield) using 2-ethoxythiophene-3-carboxylic acid (15 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.90 (s, 1H), 7.86 (d, J=6.7 Hz, 1H), 7.09 (d, J=5.9 Hz, 1H), 6.83 (d, J=5.9 Hz, 1H), 4.67 (ddd, J=12.3, 6.7, 5.3 Hz, 1H), 4.28 (q, 1=7.0 Hz, 2H), 2.77 (ddd, J=16.8, 13.3, 5.2 Hz, 1H), 2.57-2.52 (m, 1H), 2.18 (dtd, J=12.9, 5.4, 2.4 Hz, 1H), 2.01 (qd, J=12.8, 4.5 Hz, 1H), 1.45 (t, I=7.0 Hz, 3H).
LCMS (m/z [M+H]+): 283.1;
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (21% yield) using 2-methoxy-4-methylthiophene-3-carboxylic acid (10 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.83 (s, 1H), 7.97 (d, J=7.6 Hz, 1H), 6.47 (q, J=1.2 Hz, 1H), 4.66 (ddd, J=11.5, 7.6, 6.1 Hz, 1H), 3.98 (s, 3H), 2.75 (ddd, J=17.3, 12.5, 6.5 Hz, 1H), 2.52 (dd, J=8.0, 4.2 Hz, 1H), 2.23 (d, J=1.1 Hz, 3H), 2.12-1.99 (m, 2H).
LCMS (m/z [M−H]−): 281.1
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (76% yield) using 2-methoxy-5-methylthiophene-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.85 (s, 1H), 7.74 (d, J=7.5 Hz, 1H), 6.80 (q, J=1.2 Hz, 1H), 4.68 (ddd, J=11.9, 7.5, 6.0 Hz, 1H), 4.00 (s, 3H), 2.77 (ddd, J=17.3, 12.9, 6.3 Hz, 1H), 2.53 (q, J=1.5 Hz, 1H), 2.33 (d, J=1.3 Hz, 3H), 2.15-2.00 (m, 2H).
LCMS (m/z [M+H]+): 283.05
Step A: Tert-butyl N-({4-[(2,6-dioxopiperidin-3-yl)carbarmoyl]-5-methoxythiophen-2-yl}methyl)carbamate) was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (47% yield), and 5-(((tert-butoxycarbonyl)amino)methyl)-2-methoxythiophene-3-carboxylic acid (20 mg) as a starting material.
Step B: To a solution of tert-butyl N-({4-[(2,6-dioxopiperidin-3-yl)carbamoyl]-5-methoxythiophen-2-yl}methyl)carbamate) (8.7 mg, 0.022 mmol, 1 eq.) in dioxane (2 mL) was added 36% HCl (0.2 mL). The reaction was stirred at rt for 3 h and concentrated under reduced pressure to give a 5-(aminomethyl)-N-(2,6-dioxopiperidin-3-yl)-2-methoxythiophene-3-carboxamide hydrochloride (100% yield).
1H NMR (500 MHz, DMSO) δ 10.86 (s, 1H), 8.15 (s, 3H), 7.78 (d, J=7.6 Hz, 1H), 7.28 (s, 1H), 4.69 (ddd, J=12.7, 7.6, 5.5 Hz, 1H), 4.06 (s, 3H), 3.39 (s, 2H), 2.77 (ddd, J=17.2, 13.5, 5.7 Hz, 1H), 2.59-2.52 (m, 1H), 2.11 (qd, J=12.9, 4.4 Hz, 1H), 2.03 (ddt, J=10.2, 5.7, 2.9 Hz, 1H).
LCMS (m/z [M+H]+): 298.1
Step A: To a stirred solution of 3,5-dibromo-2-methoxythiophene (500.0 mg, 1.845 mmol) in toluene (9 mL) was added cyclopropyl boronic acid (206 mg, 2.399 mmol) and K3PO4 (784 mg, 3.69 mmol) in water (3 ml), the reaction mixture was purged with argon for 15 min and then Pd(PPh3)4 (320 mg, 0.277 mmol) was added. The reaction was stirred at 90° C. for 20 h, filtered through celite bed, concentrated under reduced pressure and purified by flash column chromatography to give 3-bromo-5-cyclopropyl-2-methoxythiophene (34% yield).
Step B: To a stirred solution of 3-bromo-5-cyclopropyl-2-methoxythiophene (700 mg, 3 mmol) in THF (20 mL) was added n-BuLi (1.8 M in THF) (3.4 mL, 6.005 mmol) dropwise at −78° C. Reaction mixture was stirred for 1 h at −78° C. and benzyl chloroformate (0.86 mL, 6 mmol) was added dropwise. The reaction was continued for 1 h, quenched with water, extracted with ethyl acetate and concentrated under reduced pressure. The product was purified by flash column chromatography to give benzyl 5-cyclopropyl-2-methoxythiophene-3-carboxylate (23% yield).
Step C: To a stirred solution of benzyl 5-cyclopropyl-2-methoxythiophene-3-carboxylate (350 mg, 1.215 mmol) in THF (6 mL) and methanol (6 mL) at 5-10° C. was added 50% aq. NaOH (12 ml). The reaction mixture was stirred at RT for 16 h and acidified with 6 M HCl. The solids were filtered, washed with pentane and dried to give 5-cyclopropyl-2-methoxythiophene-3-carboxylic acid (76% yield).
Step D: 5-cyclopropyl-N-(2,6-dioxopiperidin-3-yl)-2-methoxythiophene-3-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (76% yield) using 5-cyclopropyl-2-methoxythiophene-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.85 (s, 1H), 7.74 (d, J=7.5 Hz, 1H), 6.78 (d, J=1.0 Hz, 1H), 4.67 (ddd, J=12.1, 7.5, 5.8 Hz, 1H), 4.00 (s, 3H), 2.77 (ddd, J=17.3, 13.1, 6.1 Hz, 1H), 2.60-2.51 (m, 1H), 2.15-1.95 (m, 3H), 0.95-0.88 (m, 2H), 0.65-0.59 (m, 2H).
LCMS (m/z [M+H]+): 309.0
Step A: To 3,5-dibromo-2-methoxythiophene (4.0 g, 14.71 mmol) in dry THF (30 mL) was added 2.5M n-BuLi hexane solution (6.47 mL, 16.2 mmol) at −78° C. under argon atmosphere and the solution was stirred for 1 h. Tri-n-butyl borate (8.35 mL, 29.42 mmol) was added to the reaction mixture, the mixture was stirred for 1.5 h and warmed to RT. 20% Na2CO3 (33.6 mL), iodobenzene (1.65 mL, 14.71 mmol), and Pd(PPh3)4 (0.85 g, 0.73 mmol) were added and the reaction mixture was refluxed for 16 h. The reaction mixture was extracted with ether, dried over MgSO4, concentrated under reduced pressure and purified by flash column chromatography to give 3-bromo-2-methoxy-5-phenylthiophene (50% yield).
Step B: 3-Bromo-2-methoxy-5-phenylthiophene (900 mg, 3.34 mmol) was dissolved in THF (15 mL) and cooled to −78° C. 1.8M n-BuLi in hexane (3.7 mL, 6.68 mmol) was added dropwise at −78° C. Reaction mixture was stirred for 1 h at −78° C. and benzyl chloroformate (0.95 mL, 6.68 mmol) was added dropwise. The reaction was continued for 1 h, quenched with water, extracted with ethyl acetate and concentrated under reduced pressure. The product was purified by flash column chromatography to give benzyl 2-methoxy-5-phenylthiophene-3-carboxylate (23% yield).
Step C: Benzyl 2-methoxy-5-phenylthiophene-3-carboxylate (230 mg, 0.71 mmol) was dissolved in THF (5 mL). MeOH (5 mL) and 50% NaOH solution (10 mL) were added and the reaction mixture was stirred at RT for 16 h and acidified with 6 M HCl. The solids were filtered, washed with pentane and dried to give 2-methoxy-5-phenylthiophene-3-carboxylic acid (130 mg, 78%) as off white solid.
Step D: N-(2,6-dioxopiperidin-3-yl)-2-methoxy-5-phenylthiophene-3-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (71% yield) using 2-methoxy-5-phenylthiophene-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.88 (s, 1H), 7.87 (d, J=7.6 Hz, 1H), 7.63-7.57 (m, 2H), 7.51 (s, 1H), 7.48-7.38 (m, 2H), 7.33-7.27 (m, 1H), 4.73 (ddd, J=12.7, 7.6, 5.6 Hz, 1H), 4.12 (s, 3H), 2.79 (ddd, J=17.3, 13.4, 5.8 Hz, 1H), 2.57-2.52 (m, 1H), 2.19-2.03 (m, 2H).
LCMS (m/z [M+H]+): 345.2
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions D, above, (21% yield) using 5-bromo-2-methoxythiophene-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.85 (s, 1H), 7.82 (d, J=7.7 Hz, 1H), 7.19 (s, 1H), 4.77-4.61 (m, 1H), 4.04 (s, 3H), 2.76 (ddd, J=17.4, 13.5, 5.7 Hz, 1H), 2.56 (d, J=18.3 Hz, 1H), 2.14-2.04 (m, 1H), 2.01 (dtd, J=8.2, 5.6, 2.6 Hz, 1H).
LCMS (m/z [M+H]+): 346.8
Step A: To stirred solution of AlCl3 (2.1 g, 15.544 mmol) in DCM (20 mL) at −78° C. was added tert-butyl bromide (1.9 g, 13.472 mmol) in DCM (10 mL) dropwise at −78° C. and stirred for 20 min. 3-bromo-2-methoxythiophene (2 g, 10.363 mmol) in DCM (10 mL) was added dropwise stirred for 2 h. The reaction mixture was warmed to RT and stirred for another 16 h. The reaction mixture was quenched with water and extracted with DCM, concentrated under reduced pressure and purified by flash column chromatography to give 3-bromo-5-(tert-butyl)-2-methoxythiophene (31% yield).
Step B: To a stirred solution of 3-bromo-5-(tert-butyl)-2-methoxythiophene (900 mg, 3.614 mmol) in THF (22 mL) was added n-BuLi (1.8 M in THF) (4 ml, 7.229 mmol) dropwise at −78° C. Reaction mixture was stirred for 1 h at −78° C. and benzyl chloroformate (1.03 ml, 7.229 mmol) was added dropwise. The reaction was continued for 1 h, quenched with water, extracted with ethyl acetate and concentrated under reduced pressure. The product was purified by flash column chromatography to give benzyl 5-(tert-butyl)-2-methoxythiophene-3-carboxylate (220 mg, 20% yield) as light yellow oil.
Step C: To a stirred solution of benzyl 5-(tert-butyl)-2-methoxythiophene-3-carboxylate (450 mg, 1.47 mmol) in THF (8 mL) and methanol (8 mL) at 5° C. was added 50% aq. NaOH (16 mL). The reaction mixture was stirred at RT for 16 h and acidified with 6 M HCl. The solids were filtered, washed with pentane and dried to give 5-(tert-butyl)-2-methoxythiophene-3-carboxylic acid (69% yield).
Step D: 5-(tert-butyl)-N-(2,6-dioxopiperidin-3-yl)-2-methoxythiophene-3-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (75% yield) using 5-(tert-butyl)-2-methoxythiophene-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.85 (s, 1H), 7.75 (d, J=7.5 Hz, 1H), 6.83 (s, 1H), 4.68 (ddd, J=12.2, 7.5, 5.7 Hz, 1H), 4.02 (s, 3H), 2.77 (ddd, J=17.3, 13.3, 6.0 Hz, 1H), 2.57-2.52 (m, 1H), 2.17-1.99 (m, 2H), 1.31 (s, 9H).
LCMS (m/z [M+H]+): 325.2
Tert-butyl (3-((2,6-dioxopiperidin-3-yl)carbamoyl)thiophen-2-yl)carbamate (1.0 g, 2.8 mmol) was dissolved in dichloromethane (10 mL) and a 10% solution of HCl in dioxane (3 mL) was added dropwise. The reaction mixture was stirred for 48 h at room temperature. The mixture was concentrated under reduced pressure and purified by preparative HPLC to give 2-amino-N-(2,6-dioxopiperidin-3-yl)thiophene-3-carboxamide (4% yield).
1H NMR (500 MHz, DMSO) δ 10.78 (s, 1H), 7.96 (d, J=8.3 Hz, 1H), 7.22 (s, 2H), 7.07 (d, J=5.8 Hz, 1H), 6.28 (d, J=8.3 Hz, 1H), 4.69-4.61 (m, 1H), 2.82-2.68 (m, 1H), 2.57-2.52 (m, 1H), 2.15-2.04 (m, 1H), 1.96-1.84 (m, 1H).
LCMS (m/z [M+H]+): 254.0
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions A, above (24% yield), with 5-chloro-2-methoxythiophene-3-carboxylic acid as a starting material.
1H NMR (400 MHz, DMSO) δ 10.87 (s, 1H), 7.83 (d, J=7.6 Hz, 1H), 7.08 (s, 1H), 4.74-4.60 (m, 1H), 4.04 (s, 3H), 2.85-2.68 (m, 1H), 2.58-2.51 (m, 1H), 2.18-1.93 (m, 2H)
LCMS (m/z [M+H]+): 303.0
2-Nitrothiophene-3-sulfonyl chloride (3.4 g, 15 mmol) was added to a solution of 3-aminopiperidine-2,6-dione (2.06 g, 12.5 mmol, hydrochloride salt) in pyridine (20 mL) cooled to −10° C. The resulting mixture was stirred for 16 h at room temperature. The mixture was concentrated under reduced pressure, diluted with water, acidified to pH=3, and extracted with EtOAc (3×20 mL). The combined organic layers were washed with water, dried over Na2SO4, and concentrated under reduced pressure. The crude product was purified by preparative HPLC to give 0.895 g of N-(2,6-dioxopiperidin-3-yl)-2-nitrothiophene-3-sulfonamide (22% yield).
Fe (0.47 g, 8.42 mmol) was added to a solution of N-(2,6-dioxopiperidin-3-yl)-2-nitrothiophene-3-sulfonamide (0.895 g, 2.80 mmol) in acetic acid (6 mL). The reaction mixture was stirred for 3 h at 50° C. Acetic anhydride (0.315 g, 3.09 mmol) was added under room temperature and the resulting mixture was stirred for 16 h at 50° C. The mixture was concentrated under reduced pressure and the residue was purified by preparative HPLC to give 0.362 g of N-{3-[(2,6-dioxopiperidin-3-yl)sulfamoyl]thiophen-2-yl}acetamide (45% yield).
NCS (0.161 g, 1.21 mmol) was added to a solution of N-{3-[(2,6-dioxopiperidin-3-yl)sulfamoyl]thiophen-2-yl}acetamide (0.362 g, 1.09 mmol) in DMF (2 mL). The reaction mixture was stirred for 16 h at room temperature, then it was diluted with water and extracted with EtOAc (3×10 mL). The organic extracts were dried over Na2SO4, concentrated, and purified by preparative HPLC to give 0.146 g of N-{5-chloro-3-[(2,6-dioxopiperidin-3-yl)sulfamoyl]thiophen-2-yl}acetamide (37% yields).
1H NMR: (400 MHz, DMSO-d5) δ 10.90 (s, 1H), 10.33 (s, 1H), 8.37 (d, J=8.0 Hz, 1H), 7.10 (s, 1H), 4.41-4.24 (m, 1H), 2.75-2.63 (m, 1H), 2.59-2.52 (m, 1H), 2.26 (s, 3H), 2.00-1.86 (m, 2H)
LCMS (m/z [M+H]−): 365.8
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above (32% yield), and 2-(1H-pyrazol-1-yl)thiophene-3-carboxylic acid as a starting material.
1H NMR (500 MHz, DMSO) δ 10.87 (s, 1H), 8.80 (d, J=8.3 Hz, 1H), 8.39-8.36 (m, 1H), 7.75-7.70 (m, 1H), 7.41 (d, J=5.5 Hz, 1H), 7.19 (d, J=5.5 Hz, 1H) 6.52-6.46 (m, 1H), 4.73-4.64 (m, 1H), 2.83-2.71 (m, 1H), 2.57-2.51 (m, 1H), 2.13-2.02 (m, 1H), 2.01-1.93 (m, 1H).
LCMS (m/z [M+H]+): 305.1
Step A: N-chlorosuccinimide (2.2 g, 16.5 mmol) was added to a solution of 2-((tert-butoxycarbonyl)amino)thiophene-3-carboxylic acid (3.3 g, 13.6 mmol) in DMF (20 mL) and the reaction mixture was stirred at RT for 2 h. The mixture was diluted with water and filtered. The solids were washed with water and dried to give 2-{[(tert-butoxy)carbonyl]amino}-5-chlorothiophene-3-carboxylic acid (84% yield).
Step B: Tert-butyl N-{5-chloro-3-[(2,6-dioxopiperidin-3-yl)carbamoyl]thiophen-2-yl}carbamate was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions A, above (81% yield), and 2-{[(tert-butoxy)carbonyl]amino}-5-chlorothiophene-3-carboxylic acid as a starting material.
Step C: 10% HCl in dioxane (2 mL) was added dropwise to a solution of the tert-butyl N-{5-chloro-3-[(2,6-dioxopiperidin-3-yl)carbamoyl]thiophen-2-yl}carbamate (2.0 g, 5.16 mmol) in dichloromethane (15 mL) and the mixture was stirred in ultrasonic bath for 8 h, concentrated under reduced pressure and purified by HPLC to give 2-amino-5-chloro-N-(2,6-dioxopiperidin-3-yl)thiophene-3-carboxamide (15% yield).
1H NMR (400 MHz, DMSO) δ 10.81 (s, 1H), 7.97 (d, J=8.3 Hz, 1H), 7.41 (brs, 2H), 7.13 (s, 1H), 4.66-4.56 (m, 1H), 2.83-2.68 (m, 1H), 2.59-2.52 (m, 1H), 2.13-1.99 (m, 1H), 1.97-1.82 (m, 1H)
LCMS (m/z [M+H]+): 288.1
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above (55% yield), and 2-(1H-pyrrol-1-yl)thiophene-3-carboxylic acid as a starting material.
1H NMR (400 MHz, DMSO) δ 10.84 (s, 1H), 8.47 (d, J=8.3 Hz, 1H), 7.40 (d, J=5.7 Hz, 1H), 7.19 (d, J=5.7 Hz, 1H) 7.11-7.01 (m, 2H), 6.28-6.12 (m, 2H), 4.70-4.59 (m, 1H), 2.84-2.65 (m, 1H), 2.58-2.52 (m, 1H), 2.09-1.85 (m, 2H).
LCMS (m/z [M+H]+): 304.1
Step A: Triethylamine (0.397 g, 3.92 mmol) and acetic anhydride (0.400 g, 3.92 mmol) were added to a solution of ethyl 2-amino-5-(trifluoromethyl)thiophene-3-carboxylate (0.852 g, 3.56 mmol) in MeCN (15 mL). The reaction mixture was stirred overnight at 50° C., cooled to rt, concentrated under reduced pressure, and extracted with DCM, dried over Na2SO4, and concentrated to give ethyl 2-acetamido-5-(trifluoromethyl)thiophene-3-carboxylate (91% yield).
Step B: 10% solution of LiOH (0.081 g, 3.4 mmol) was added to a solution of ethyl 2-acetamido-5-(trifluoromethyl)thiophene-3-carboxylate (0.911 g, 3.24 mmol) in THF (15 mL) and the resulting mixture was stirred for 5 days at RT. The solvents were evaporated under reduced pressure, the residue was diluted with water and washed with MTBE. The aqueous layer acidified by citric acid and the precipitate was filtered, washed with water, and dried to give 2-acetamido-5-(trifluoromethyl)thiophene-3-carboxylic acid (28% yield).
Step C: N-(2,6-dioxopiperidin-3-yl)-2-acetamido-5-(trifluoromethyl)thiophene-3-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions A, above (26% yield), and 2-acetamido-5-(trifluoromethyl)thiophene-3-carboxylic acid as a starting material.
1H NMR (400 MHz, DMSO) δ 11.98 (s, 1H), 10.93 (s, 1H), 9.00-8.81 (m, 1H), 8.13 (s, 1H), 4.87-4.62 (m, 1H), 2.98-2.65 (m, 2H), 2.28 (s, 3H), 2.20-2.08 (m, 1H), 2.07-1.88 (m, 1H).
LCMS (m/z [M+H]+): 364.2
Step A: A solution of methyl 4-chloroacetoacetate (13.3 g, 88.3 mmol) in THF (30 mL) was added dropwise to a suspension of 60% NaH (4.45 g, 111 mmol) in THF (150 mL) at 0° C. After addition was completed, the reaction mixture was warmed to RT and stirred for 20 min. Then, the reaction mixture was cooled to 0° C. and a solution of acetyl isothiocyanate (8.92 g, 88.2 mmol) in THF (30 mL) was added dropwise. After the addition was completed, the reaction mixture was quenched at 0° C. by NH4Cl solution, concentrated under reduced pressure and diluted with water and ethyl acetate. The mixture was filtered and the solids were dried to give methyl 2-acetamido-4-oxo-4,5-dihydrothiophene-3-carboxylate (4% yield).
Step B: POCl3 (1.05 g, 6.85 mmol) was added to a suspension of methyl 2-acetamido-4-oxo-4,5-dihydrothiophene-3-carboxylate (0.74 g, 3.44 mmol) in dioxane (10 mL) and refluxed for 2 h. The reaction mixture was cooled and poured into iced water, the product was extracted with EtOAc, dried over Na2SO4 and concentrated under reduced pressure to give methyl 4-chloro-2-acetamidothiophene-3-carboxylate (26% yield).
Step C: 4-chloro-2-acetamidothiophene-3-carboxylic acid was synthesized using Example Method 2, above, with methyl 4-chloro-2-acetamidothiophene-3-carboxylate as a starting material.
Step D: 4-chloro-N-(2,6-dioxopiperidin-3-yl)-2-acetamidothiophene-3-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above (17% yield), and 4-chloro-2-acetamidothiophene-3-carboxylic acid as a starting material.
1H NMR (400 MHz, DMSO) δ 11.13 (s, 1H), 11.07 (s, 1H), 8.54 (d, J=8.2 Hz 1H), 7.09 (s, 1H), 4.93-4.79 (m, 1H), 2.89-2.74 (m, 1H), 2.62-2.52 (m, 1H), 2.19 (s, 3H), 2.15-2.01 (m, 2H).
LCMS (m/z [M−1-1]+): 330.0
Step A: methyl 5-cyclopropyl-2-acetamidothiophene-3-carboxylate was synthesized in 69% yield using Example Method 3, above, using 2-amino-5-cyclopropylthiophene-3-carboxylate as a starting material.
Step B: 5-cyclopropyl-2-acetamidothiophene-3-carboxylic acid was synthesized in 57% yield using Example Method 2, above, and methyl 5-cyclopropyl-2-acetamidothiophene-3-carboxylate as a starting material.
Step C: 5-cyclopropyl-N-(2,6-dioxopiperidin-3-yl)-2-acetamidothiophene-3-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above (56% yield), and 5-cyclopropyl-2-acetamidothiophene-3-carboxylic acid as a starting material.
1H NMR (400 MHz, DMSO) δ 11.73 (s, 1H), 10.89 (s, 1H), 8.52 (d, J=8.5 Hz, 1H), 7.09 (s, 1H), 4.76-4.64 (m, 1H), 2.85-2.71 (m, 1H), 2.62-2.53 (m, 1H), 2.22-2.04 (m, 4H), 2.04-1.89 (m, 2H), 1.00-0.87 (m, 2H), 0.68-0.57 (m, 2H)
LCMS (m/z [M+H]+): 336.2
Step A: methyl 2-benzamido-5-chlorothiophene-3-carboxylate was synthesized using Example
Method 1, above (69% yield), using methyl 2-benzamidothiophene-3-carboxylate as a starting material.
Step B: 2-benzamido-5-chlorothiophene-3-carboxylic acid was synthesized using Example Method 2, above (70% yield), using methyl 2-benzamido-5-chlorothiophene-3-carboxylate as a starting material.
Step C: 2-benzamido-5-chloro-N-(2,6-dioxopiperidin-3-yl)thiophene-3-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions A, above (50% yield), and 2-benzamido-5-chlorothiophene-3-carboxylic acid as a starting material.
1H NMR (500 MHz, DMSO) δ 13.20 (s, 1H), 10.92 (s, 1H), 8.86-8.73 (m, 1H), 8.00-7.84 (m, 2H), 7.72-7.50 (m, 4H), 4.95-4.81 (m, 1H), 2.89-2.74 (m, 1H), 2.67-2.56 (m, 1H), 2.22-2.08 (m, 1H), 2.06-1.93 (m, 1H)
LCMS (m/z [M+H]+): 392.2
Step A: Methyl 5-chloro-2-(2-phenylacetamido)thiophene-3-carboxylate was synthesized using Example Method 1, above (75% yield), using methyl 2-(2-phenylacetamido)thiophene-3-carboxylate as a starting material.
Step B: 5-chloro-2-(2-phenylacetamido)thiophene-3-carboxylic acid was synthesized using Example Method 2, above (82% yield), using methyl 5-chloro-2-(2-phenylacetamido)thiophene-3-carboxylate as a starting material.
Step C: 5-chloro-N-(2,6-dioxopiperidin-3-yl)-2-(2-phenylacetamido)thiophene-3-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions A, above (15% yield), with 5-chloro-2-(2-phenylacetamido)thiophene-3-carboxylic acid as a starting material.
1H NMR (400 MHz, DMSO) δ 11.94 (s, 1H), 10.91 (s, 1H), 8.72-8.54 (m, 1H), 7.66-7.45 (m, 2H), 7.43-7.21 (m, 4H), 4.82-4.64 (m, 1H), 3.90 (s, 2H), 2.89-2.71 (m, 1H), 2.61-2.53 (m, 1H), 2.17-2.02 (m, 1H), 2.01-1.88 (m, 1H)
LCMS (m/z [M+H]+): 406.2
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (53% yield) using 2-cyclopropylfuran-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.83 (s, 1H), 8.24 (d, J=8.5 Hz, 1H), 7.42 (d, J=2.0 Hz, 1H), 6.83 (d, J=2.1 Hz, 1H), 4.70 (ddd, J=12.4, 8.3, 5.3 Hz, 1H), 2.87 (tt, J=8.5, 5.2 Hz, 1H), 2.78 (ddd, J=17.4, 13.4, 5.6 Hz, 1H), 2.60-2.51 (m, 1H), 2.10 (qd, J=13.0, 4.4 Hz, 1H), 1.98-1.92 (m, 1H), 1.00-0.95 (m, 2H), 0.91-0.87 (m, 2H).
LCMS (m/z [M+H]+): 263.2
Step A: To a solution of tert-butyl (3-bromofuran-2-yl)carbamate (2 g, 7.6 mmol) in DMF (40 ml) was added sodium hydride (0.28 g, 11.5 mmol) at 0° C. under nitrogen and the reaction mixture was stirred at RT for 1 h. It was then re-cooled at 0° C., methyl iodide (1.42 ml, 23 mmol) was added and the reaction mixture was stirred for an additional 1 h at RT. The reaction mixture was diluted with ethyl acetate, washed with water and brine, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give tert-butyl (3-bromofuran-2-yl)(methyl)carbamate 1.27 g (60% yield).
Step B: n-butyllithium (3.37 ml, 5.43 mmol, 1.6 M in hexane) was added slowly to a THF (30 ml) solution of tert-butyl (3-bromofuran-2-yl)(methyl)carbamate (1.5 g, 5.43 mmol) at −78° C. under nitrogen. After 15 min of stirring, a stream of dry CO2 was bubbled into the solution for 30 min. The reaction was quenched with 1M HCl (10 ml), extracted with DCM, fried over Na2SO4, concentrated under reduced pressure to give 2-((tert-butoxycarbonyl)(methyl)amino)furan-3-carboxylic acid 500 mg (38% yield).
Step C: tert-butyl (3-((2,6-dioxopiperidin-3-yl)carbamoyl)furan-2-yl)(methyl)carbamate was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above, (59% yield) using 2-((tert-butoxycarbonyl)(methyl)amino)furan-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.84 (s, 1H), 8.28 (d, J=8.3 Hz, 1H), 7.52 (d, J=2.3 Hz, 1H), 6.89 (d, J=2.3 Hz, 1H), 4.70 (ddd, J=12.2, 8.2, 5.3 Hz, 1H), 3.03 (s, 3H), 2.78 (ddd, J=17.3, 13.3, 5.5 Hz, 1H), 2.54 (dd, J=4.5, 2.9 Hz, 1H), 2.13-2.00 (m, 1H), 1.94 (dtd, J=10.2, 5.5, 2.9 Hz, 1H), 1.45-1.19 (m, 9H).
LCMS (m/z [M−H]−): 350.3
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (24% yield) using 5-[(tert-butoxycarbonylamino)methyl]thiophene-2-carboxylic acid (23 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.85 (s, 1H), 8.68 (d, J=8.4 Hz, 1H), 7.59 (d, J=3.7 Hz, 1H), 6.94 (d, J=3.7 Hz, 1H), 4.70 (ddd, J=12.7, 8.4, 5.3 Hz, 1H), 4.27 (d, J=6.0 Hz, 2H), 2.78 (ddd, J=17.4, 13.3, 5.5 Hz, 1H), 2.52-2.51 (m, 2H), 2.15-2.04 (m, 1H), 2.00-1.92 (m, 1H), 1.40 (s, 9H).
LCMS (m/z [M−H]−): 366.1
To the suspension of tert-butyl ((5-((2,6-dioxopiperidin-3-yl)carbamoyl)thiophen-2-yl)methyl)carbamate (5 mg, 0.014 mmol, 1 eq) in DCM (0.1 mL) was added TFA (20 μL) and mixture was stirred for 2 days at RT. The reaction mixture was evaporated under reduced pressure and suspended in ACN, then 4N HCl in 1,4-dioxane was added (50 μL). Concentration under reduced pressure gave 5-(aminomethyl)-N-(2,6-dioxopiperidin-3-yl)thiophene-2-carboxamide hydrochloride (92% yield).
1H NMR (500 MHz, DMSO) δ 10.88 (s, 1H), 8.86 (d, J=8.4 Hz, 1H), 8.29 (s, 3H), 7.71 (d, J=3.8 Hz, 1H), 7.25 (d, J=3.8 Hz, 1H), 4.77-4.65 (m, 1H), 4.27 (s, 2H), 2.79 (ddd, J=17.4, 13.4, 5.5 Hz, 1H), 2.52-2.51 (m, 1H), 2.12 (qd, J=13.2, 4.7 Hz, 1H), 1.97 (dddd, J=10.9, 8.2, 5.4, 2.8 Hz, 1H).
LCMS (m/z [M−1-1]+): 268.0
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (31% yield) using 4-((tert-butoxycarbonyl)aminomethyl)thiophene-2-carboxylic acid (37.8 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.85 (s, 1H), 8.77 (d, J=8.4 Hz, 1H), 7.65 (d, J=1.3 Hz, 1H), 7.47 (d, J=1.2 Hz, 1H), 4.71 (ddd, J=12.6, 8.4, 5.3 Hz, 1H), 4.09 (d, J=5.9 Hz, 2H), 2.78 (ddd, J=17.4, 13.3, 5.5 Hz, 1H), 2.52-2.51 (m, 2H), 2.16-2.04 (m, 1H), 1.96 (dddd, J=10.9, 8.2, 5.4, 2.9 Hz, 1H), 1.39 (s, 9H).
LCMS (m/z [M−H]−): 366.2
To the suspension of tert-butyl ((5-((2,6-dioxopiperidin-3-yl)carbamoyl)thiophen-3-yl)methyl)carbamate (15.8 mg, 0.043 mmol) in DCM (0.5 mL) was added TFA (0.1 mL) and mixture was stirred for 2 days at RT. Mixture was evaporated and purified by HPLC to give 4-(aminomethyl)-N-(2,6-dioxopiperidin-3-yl)thiophene-2-carboxamide formate (63% yield).
1H NMR (500 MHz, DMSO) δ 10.85 (s, 1H), 8.78 (d, J=8.4 Hz, 1H), 7.78 (d, J=1.2 Hz, 1H), 7.71 (s, 1H), 4.73 (ddd, J=12.6, 8.4, 5.4 Hz, 1H), 3.93 (s, 2H), 2.79 (ddd, J=17.4, 13.3, 5.5 Hz, 1H), 2.56 (dd, J=4.4, 3.0 Hz, 1H), 2.52 (dd, J=4.3, 2.5 Hz, 1H), 2.11 (qd, J=12.9, 4.5 Hz, 1H), 1.96 (dddd, J=10.9, 8.2, 5.4, 2.9 Hz, 1H).
LCMS (m/z [M+H]+): 268.1
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions D, above, (20% yield) using 4-hydroxythiophene-2-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.86 (s, 1H), 9.75 (s, 1H), 8.67 (d, J=8.4 Hz, 1H), 7.35 (d, J=1.7 Hz, 1H), 6.55 (d, J=1.7 Hz, 1H), 4.71 (ddd, J=12.5, 8.4, 5.4 Hz, 1H), 2.85-2.73 (m, 1H), 2.58-2.52 (m, 1H), 2.10 (qd, J=13.0, 4.5 Hz, 1H), 2.02-1.92 (m, 1H).
LCMS (m/z [M−H]−): 253.1
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions D, above, (32% yield) using 4-methoxythiophene-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.89 (s, 1H), 8.12 (d, J=7.4 Hz, 1H), 8.09 (d, J=3.6 Hz, 1H), 6.81 (d, J=3.6 Hz, 1H), 4.72 (dt, J=10.3, 7.5 Hz, 1H), 3.89 (s, 3H), 2.85-2.72 (m, 1H), 2.54 (t, J=3.5 Hz, 1H), 2.15-2.05 (m, 2H).
LCMS (m/z [M+H]+): 269.0
Step A: (4-Bromo-3-methoxythiophen-2-yl)methanol (0.8 g, 3.58 mmol) was dissolved in DCM (10 mL). TBDMSCl (1.08 g, 7.17 mmol) and imidazole (0.6 g, 8.96 mmol) were added and the reaction mixture was stirred at RT for 48 h, diluted, washed with water, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give ((4-bromo-3-methoxythiophen-2-yl)methoxy)(tert-butyl)dimethylsilane (82% yield).
Step B: ((4-bromo-3-methoxythiophen-2-yl)methoxy)(tert-butyl)dimethylsilane (1.5 g, 4.45 mmol) was dissolved in THF (20 mL) and cooled to −78° C. n-BuLi (3.7 mL, 6.67 mmol) was added dropwise and the reaction mixture was stirred for 30 min. Methyl chloroformate (0.62 mL, 8.0 mmol) was added and stirring was continued for 2 h at −78° C. Ammonium chloride solution was added, the product was extracted with ethyl acetate, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give methyl 5-(((tert-butyldimethylsilyl)oxy)methyl)-4-methoxythiophene-3-carboxylate (51% yield).
Step C: methyl 5-(((tert-butyldimethylsilyl)oxy)methyl)-4-methoxythiophene-3-carboxylate (0.8 g, 2.52 mmol) was dissolved in THF (10 mL) and TBAF (1M solution in THF) (5.06 mL, 5.0 mmol) was added at 0° C. Reaction mixture was stirred at rt for 4 h, diluted with ethyl acetate and washed with water. Organic phase was dried over Na2SO4 and concentrated under reduced pressure to give methyl 5-(hydroxymethyl)-4-methoxythiophene-3-carboxylate (88% yield).
Step D: methyl 5-(hydroxymethyl)-4-methoxythiophene-3-carboxylate (0.2 g, 1.0 mmol) was dissolved in toluene (3 mL) and cooled to 0° C. DBU (0.19 mL, 1.3 mmol), and DPPA (0.26 mL, 1.2 mmol) were added and reaction mixture was stirred at RT for 16 h. The mixture was diluted with ethyl acetate, washed with water, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give methyl 5-(azidomethyl)-4-methoxythiophene-3-carboxylate (78% yield).
Step E: methyl 5-(azidomethyl)-4-methoxythiophene-3-carboxylate (40 mg, 0.176 mmol) was dissolved in MeOH (5 mL) and 10% Pd/C (20 mg) was added. Reaction mixture was stirred under H2 atmosphere at RT for 3 h, filtered through celite bed and concentrated under reduced pressure to give methyl 5-(aminomethyl)-4-methoxythiophene-3-carboxylate that was used directly in the next step.
Step F: methyl 5-(aminomethyl)-4-methoxythiophene-3-carboxylate (215 mg, 1.06 mmol) was dissolved in a dioxane-water (1:1; 6 mL). triethylamine (0.22 mL, 1.6 mmol) and Boc2O (0.29 mL, 1.28 mmol) were added and the reaction mixture was stirred at RT for 18 h, diluted with ethyl acetate, washed with water, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give methyl 5-(((tert-butoxycarbonyl)amino)methyl)-4-methoxythiophene-3-carboxylate (38% yield, 2 steps).
Step G: To 5-(((tert-butoxycarbonyl)amino)methyl)-4-methoxythiophene-3-carboxylate (200.0 mg, 0.66 mmol) in THF (1.0 mL) was added methanol (1.0 mL) and 50% aqueous NaOH (2 mL), the reaction mixture was stirred at RT for 16 h, diluted with water and acidified with citric acid. The product was extracted with ethyl acetate, concentrated and triturated with diethyl ether to give 5-(((tert-butoxycarbonyl)amino)methyl)-4-methoxythiophene-3-carboxylic acid (83% yield).
Step H: tert-butyl ((4-((2,6-dioxopiperidin-3-yl)carbamoyl)-3-methoxythiophen-2-yl)methyl)carbamate was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above (23% yield), and 5-(((tert-butoxycarbonyl)amino)methyl)-4-methoxythiophene-3-carboxylic acid (30 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.86 (s, 1H), 8.67 (d, J=8.4 Hz, 1H), 7.64 (s, 1H), 7.42 (t, J=5.8 Hz, 1H), 4.79-4.63 (m, 1H), 4.15 (d, J=6.0 Hz, 2H), 3.80 (s, 3H), 2.79 (ddd, J=17.4, 13.2, 5.5 Hz, 1H), 2.60-2.52 (m, 1H), 2.08 (qd, J=12.9, 4.4 Hz, 1H), 2.03-1.90 (m, 1H), 1.39 (s, 9H).
LCMS (m/z [M−H]−): 396.0
Step I: tert-butyl ((4-((2,6-dioxopiperidin-3-yl)carbamoyl)-3-methoxythiophen-2-yl)methyl) carbamate (5 mg) was dissolved in 2 mL of TFA and the solution was stirred at RT for 2 h. The volatiles were removed under reduced pressure to give 5-(aminomethyl)-N-(2,6-dioxopiperidin-3-yl)-4-methoxythiophene-3-carboxamide trifluoroacetate (87% yield).
1H NMR (500 MHz, DMSO) δ 10.88 (s, 1H), 8.88 (d, J=8.3 Hz, 1H), 8.18 (s, 3H), 7.77 (s, 1H), 4.79-4.65 (m, 1H), 4.10 (q, J=5.4 Hz, 2H), 2.79 (ddd, J=17.5, 13.3, 5.5 Hz, 1H), 2.54 (ddd, J=9.0, 6.1, 2.2 Hz, 1H), 2.17-2.07 (m, 1H), 2.03-1.94 (m, 1H).
Step A: methyl 2-((tert-butoxycarbonyl)amino)thiophene-3-carboxylate (100 mg, 1 eq.) was added to a stirred mixture of 60% NaH (1.2 eq.) suspended in mineral oil in dry DMF in an inert atmosphere, next was added Mel (1.2 eq.). The resulting mixture was stirred at room temperature for 18 hours, solvent was removed under reduced pressure and the residue was purified by flash column chromatography to give methyl 2-((tert-butoxycarbonyl)(methyl)amino)thiophene-3-carboxylate (49% yield).
Step B: 1M NaOH in H2O (10 eq.) was added to a solution of methyl 2-((tert-butoxycarbonyl)(methyl)amino)thiophene-3-carboxylate (52.0 mg, 1. eq.) in methanol and was stirring at room temperature for 18 hours. When the reaction was completed, to the acidified with 1M HCl, concentrated under reduced pressure and partitioned between ethyl acetate and water. Organic layer was washed with brine, dried over Na2SO4 and evaporated. 2-((tert-butoxycarbonyl)(methyl)amino)thiophene-3-carboxylic acid (100%) was used without further purification in the next step.
Step C: tert-butyl (3-((2,6-dioxopiperidin-3-yl)carbamoyl)thiophen-2-yl)(methyl)carbamate was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (57% yield) using 2-((tert-butoxycarbonyl)(methyl)amino)thiophene-3-carboxylic acid (49.4 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.84 (s, 1H), 8.30 (d, J=8.2 Hz, 1H), 7.38 (d, I=5.8 Hz, 1H), 7.25 (d, J=5.9 Hz, 1H), 4.70 (ddd, J=13.0, 8.1, 5.3 Hz, 1H), 3.10 (s, 3H), 2.78 (ddd, J=17.3, 13.3, 5.6 Hz, 1H), 2.59-2.53 (m, 1H), 2.11-2.01 (m, 1H), 1.95 (dtd, J=12.8, 5.4, 2.9 Hz, 1H), 1.32 (s, 9H).
LCMS (m/z [M-Boc+H]+): 268.0
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions D, above, (50% yield) using 5-phenylthiophene-3-carboxylic acid (16.7 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.87 (s, 1H), 8.64 (d, J=8.4 Hz, 1H), 8.15 (d, J=1.4 Hz, 1H), 7.89 (d, J=1.4 Hz, 1H), 7.71-7.65 (m, 2H), 7.51-7.42 (m, 2H), 7.40-7.33 (m, 1H), 4.83-4.73 (m, 1H), 2.81 (ddd, J=18.6, 13.2, 5.6 Hz, 1H), 2.60-2.53 (m, 1H), 2.18-2.04 (m, 1H), 2.00 (ddt, J=11.0, 8.4, 4.2 Hz, 1H).
LCMS (m/z [M]+): 315.0
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (88% yield) using 2-acetamido-5-phenylthiophene-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 11.84 (s, 1H), 10.92 (s, 1H), 8.72 (d, J=8.2 Hz, 1H), 7.87 (s, 1H), 7.62-7.55 (m, 2H), 7.48-7.39 (m, 2H), 7.35-7.26 (m, 1H), 4.83-4.70 (m, 1H), 2.81 (ddd, J=17.2, 13.4, 5.5 Hz, 1H), 2.58 (dt, J=16.8, 3.8 Hz, 1H), 2.24 (s, 3H), 2.17 (qd, J=12.9, 4.5 Hz, 1H), 2.01 (dtd, J=12.9, 5.4, 2.8 Hz, 1H).
LCMS (m/z [M+H]+): 372.3
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (83% yield) using 2-benzamido-5-phenylthiophene-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 13.21 (s, 1H), 10.96 (s, 1H), 8.87 (d, J=8.4 Hz, 1H), 7.99 (s, 1H), 7.97-7.91 (m, 2H), 7.74-7.68 (m, 1H), 7.68-7.61 (m, 4H), 7.51-7.43 (m, 2H), 7.38-7.29 (m, 1H), 4.94 (ddd, J=13.0, 8.3, 5.3 Hz, 1H), 2.90-2.79 (m, 1H), 2.60 (dt, J=17.7, 3.8 Hz, 1H), 2.23-2.12 (m, 1H), 2.07-2.02 (m, 1H).
LCMS (m/z [M+H]+): 434.2
Step A: To the solution of ethyl 5-phenyl-2-(2-(pyrrolidin-1-yl)acetamido)thiophene-3-carboxylate (20 mg, 0.056 mmol) in EtOH (0.6 mL) was added H2O (0.1 mL) followed by NaOH (4 eq). Reaction was stirred at 50° C. for 3 h. The reaction mixture was acidified with 1M HCl, concentrated under reduced pressure and the crude 5-phenyl-2-(2-(pyrrolidin-1-yl)acetamido)thiophene-3-carboxylic acid was used directly in the next step (98% yield).
Step B: N-(2,6-dioxopiperidin-3-yl)-5-phenyl-2-(2-(pyrrolidin-1-yl)acetamido)thiophene-3-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (15% yield) using 5-phenyl-2-(2-(pyrrolidin-1-yl)acetamido)thiophene-3-carboxylic acid (18 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 12.51 (s, 1H), 10.91 (s, 1H), 8.61 (d, J=8.5 Hz, 1H), 8.14 (s, 1H), 7.88 (s, 1H), 7.62-7.57 (m, 2H), 7.47-7.41 (m, 2H), 7.33-7.28 (m, 1H), 4.87 (ddd, J=12.4, 8.5, 5.5 Hz, 1H), 3.41 (s, 2H), 2.91-2.75 (m, 1H), 2.63 (td, J=4.9, 4.4, 2.3 Hz, 4H), 2.57 (ddd, J=17.3, 4.4, 2.9 Hz, 1H), 2.16-2.06 (m, 1H), 2.06-1.98 (m, 1H), 1.82-1.74 (m, 4H).
LCMS (m/z [M−1-1]1: 441.1
Step A: To the solution of methyl 2-(((tert-butoxycarbonyl)amino)methyl)thiophene-3-carboxylate (35 mg, 0.129 mmol, 1. eq.) in THF (1.0 mL) was added H2O (0.3 mL) followed by NaOH (6 eq). Reaction was stirred at RT for 18 h and at 50° C. for 4 h. The reaction mixture was acidified with 1M HCl, extracted with EtOAc, dried over Na2SO4 and concentrated to give 2-(((tert-butoxycarbonyl)amino)methyl)thiophene-3-carboxylic acid (86% yield).
Step B: Tert-butyl ((3-((2,6-dioxopiperidin-3-yl)carbamoyl)thiophen-2-yl)methyl)carbamate was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (37% yield) using 2-(((tert-butoxycarbonyl)amino)methyl)thiophene-3-carboxylic acid (30 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.84 (s, 1H), 8.47 (d, J=8.3 Hz, 1H), 7.39 (s, 2H), 4.78-4.67 (m, 1H), 4.57 (d, J=6.7 Hz, 2H), 2.78 (ddd, J=17.4, 13.3, 5.5 Hz, 1H), 2.57-2.51 (m, 2H), 2.10 (qd, J=13.0, 4.5 Hz, 1H), 2.03-1.91 (m, 1H), 1.40 (s, 9H).
LCMS (m/z [M−H]−): 366.0
To the suspension of tert-butyl ((3-((2,6-dioxopiperidin-3-yl)carbamoyl)thiophen-2-yl)methyl)carbamate (12.7 mg, 0.035 mmol, 1 eq) in DCM (0.2 mL) was added TFA (50 μl) and mixture was stirred for 2 days at RT. The reaction mixture was evaporated under reduced pressure and suspended in ACN, then 4N HCl in 1,4-dioxane was added (50 μL). Concentration under reduced pressure gave 2-(aminomethyl)-N-(2,6-dioxopiperidin-3-yl)thiophene-3-carboxamide hydrochloride (98% yield).
1H NMR (500 MHz, DMSO) δ 10.91 (s, 1H), 8.90 (d, J=8.3 Hz, 1H), 8.25 (s, 3H), 7.68 (d, J=5.4 Hz, 1H), 7.59 (d, J=5.4 Hz, 1H), 4.82-4.70 (m, 1H), 2.80 (ddd, J=17.4, 13.4, 5.5 Hz, 1H), 2.60-2.53 (m, 1H), 2.15 (qd, J=13.0, 4.5 Hz, 1H), 1.98 (dddd, J=10.8, 8.2, 5.3, 2.9 Hz, 1H).
LCMS (m/z [M+H]+): 268.0
To the mixture of 2-methoxythiophene-3-carboxylic acid (20.0 mg, 0.126 mmol, 1.000 eq) and CDI (30.8 mg, 0.189 mmol, 1.500 eq) was added DMF (1.0 mL) and the reaction mixture was stirred for 1 h at 50° C. After cooling to RT 3-aminopyrrolidine-2,5-dione hydrochloride (22.8 mg, 0.151 mmol, 1.200 eq) was added and the reaction mixture was stirred overnight. The solvent was removed under reduced pressure and the residue was purified by preparative TLC (78% yield).
1H NMR (500 MHz, DMSO) δ 11.19 (s, 1H), 8.14 (d, J=7.7 Hz, 1H), 7.08 (d, J=5.9 Hz, 1H), 6.83 (d, J=6.0 Hz, 1H), 4.65 (ddd, J=9.3, 7.8, 5.8 Hz, 1H), 4.05 (s, 3H), 2.89 (dd, J=17.4, 9.4 Hz, 1H), 2.60 (dd, J=17.4, 5.8 Hz, 1H).
LCMS (m/z [M+H]+): 254.8
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions D, above, (72% yield) using 2-(cyclopropyl)thiophene-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.83 (s, 1H), 8.32 (d, J=8.3 Hz, 1H), 7.27 (d, J=5.4 Hz, 1H), 7.24 (d, J=5.4 Hz, 1H), 4.76-4.68 (m, 1H), 2.96 (tt, I=8.4, 5.2 Hz, 1H), 2.78 (ddd, J=17.4, 13.3, 5.5 Hz, 1H), 2.57-2.51 (m, 1H), 2.11 (qd, J=13.0, 4.5 Hz, 1H), 1.98 (dddd, J=11.0, 8.3, 5.4, 2.9 Hz, 1H), 1.10 (ddd, J=8.4, 6.0, 4.0 Hz, 2H), 0.68-0.63 (m, 2H).
LCMS (m/z [M+H]+): 279.1
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (45% yield) using 5-methoxy-1,3-oxazole-4-carboxylic acid (7 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.80 (s, 1H), 8.11 (d, J=8.4 Hz, 1H), 8.01 (s, 1H), 4.67 (ddd, J=12.5, 8.4, 5.3 Hz, 1H), 4.12 (s, 3H), 2.81-2.73 (m, 1H), 2.58 (s, 1H), 2.20-2.09 (m, 1H), 1.97-1.89 (m, 1H).
LCMS (m/z [M+H]+): 254.0
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions D, above, (86% yield) using 5-ethoxy-2-methyloxazole-4-carboxylic acid (30 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.79 (s, 1H), 8.04 (d, J=8.3 Hz, 1H), 4.69-4.60 (m, 1H), 4.44 (q, J=7.1 Hz, 2H), 2.75 (ddd, J=17.4, 13.7, 5.5 Hz, 1H), 2.48 (d, J=2.8 Hz, 1H), 2.33 (s, 3H), 2.13 (qd, J=12.9, 4.5 Hz, 1H), 1.93 (dddd, J=10.8, 7.9, 5.4, 2.5 Hz, 1H), 1.32 (t, J=7.1 Hz, 3H).
LCMS (m/z [M+H]+): 282.1
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (39% yield), and 5-methoxy-2-phenyloxazole-4-carboxylic acid (9.4 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.83 (s, 1H), 8.18 (d, J=8.4 Hz, 1H), 7.97-7.92 (m, 2H), 7.58-7.50 (m, 3H), 4.73 (ddd, J=12.5, 8.4, 5.3 Hz, 1H), 4.23 (s, 3H), 2.84-2.73 (m, 1H), 2.59-2.52 (m, 1H), 2.18 (qd, J=12.9, 4.4 Hz, 1H), 1.97 (dtd, J=12.9, 5.4, 2.6 Hz, 1H).
LCMS (m/z [M+H]+): 330.0
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions D, above, (27% yield) using 5-(morpholin-4-yl)-2-phenyl-1,3-oxazole-4-carboxylic acid (8 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.82 (s, 1H), 8.15 (d, J=8.2 Hz, 1H), 7.94-7.88 (m, 2H), 7.56-7.45 (m, 3H), 4.67 (ddd, J=12.8, 8.1, 5.3 Hz, 1H), 3.73 (s, 8H), 2.76 (ddd, J=17.2, 13.6, 5.4 Hz, 1H), 2.58 (d, J=4.9 Hz, 1H), 2.18 (qd, J=12.9, 4.4 Hz, 1H), 2.03-1.93 (m, 1H).
LCMS (m/z [M+H]+): 385.1
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (31% yield) using 2-((tert-butoxycarbonyl)amino)thiazole-5-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 11.73 (s, 1H), 10.86 (s, 1H), 8.70 (d, J=8.4 Hz, 1H), 8.00 (s, 1H), 4.78-4.66 (m, 1H), 2.78 (ddd, J=17.5, 13.2, 5.6 Hz, 1H), 2.52-2.51 (m, 1H), 2.07 (qd, J=12.9, 4.5 Hz, 1H), 1.97 (dddd, J=11.3, 8.4, 5.5, 2.9 Hz, 1H), 1.50 (s, 9H).
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (29% yield) using 2-(trifluoromethyl)thiazole-5-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.87 (s, 1H), 7.89 (d, J=7.4 Hz, 1H), 7.75 (d, J=5.5 Hz, 1H), 7.14 (d, J=5.5 Hz, 1H), 4.78-4.63 (m, 1H), 3.98 (s, 3H), 3.31 (s, 3H), 2.77 (ddd, J=17.4, 13.1, 6.3 Hz, 1H), 2.54-2.51 (m, 1H), 2.17-2.04 (m, 2H).
LCMS (m/z [M−H]−): 305.8
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (83% yield) using 2-(tert-butyl)thiazole-5-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.88 (s, 1H), 8.88 (d, J=8.4 Hz, 1H), 8.26 (s, 1H), 4.73 (ddd, J=13.4, 8.3, 5.4 Hz, 1H), 2.79 (ddd, J=17.5, 13.3, 5.6 Hz, 1H), 2.58-2.51 (m, 1H), 2.15-2.03 (m, 1H), 1.98 (ddd, J=12.9, 5.4, 2.8 Hz, 1H), 1.39 (s, 9H).
LCMS (m/z [M+H]+): 295.9
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (68% yield) using 2-((((benzyloxy)carbonyl)amino)methyl)thiazole-5-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.88 (s, 1H), 8.91 (d, J=8.3 Hz, 1H), 8.28 (s, 1H), 8.25 (t, J=5.9 Hz, 1H), 7.38 (d, J=4.1 Hz, 3H), 7.33 (dd, J=8.8, 4.5 Hz, 1H), 5.09 (s, 2H), 4.78-4.69 (m, 1H), 4.48 (d, J=6.1 Hz, 2H), 2.79 (ddd, J=18.6, 13.3, 5.6 Hz, 1H), 2.52 (d, J=1.9 Hz, 1H), 2.09 (qd, J=12.9, 4.4 Hz, 1H), 1.99 (tdd, J=8.2, 5.4, 2.8 Hz, 1H).
LCMS (m/z [M+H]+): 403.1
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (16% yield) using 2-methoxythiazole-5-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.87 (s, 1H), 8.79 (d, J=8.3 Hz, 1H), 7.87 (s, 1H), 4.71 (ddd, J=12.6, 8.3, 5.4 Hz, 1H), 4.06 (s, 3H), 2.78 (ddd, J=17.5, 13.2, 5.6 Hz, 1H), 2.53-2.51 (m, 1H), 2.07 (qd, J=12.9, 4.5 Hz, 1H), 1.97 (dddd, J=11.0, 8.3, 5.5, 2.8 Hz, 1H).
LCMS (m/z [M+H]+): 269.9
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (43% yield) using 2-(methylthio)thiazole-5-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.88 (s, 1H), 8.90 (d, J=8.3 Hz, 1H), 8.26 (s, 1H), 4.73 (ddd, J=12.6, 8.3, 5.4 Hz, 1H), 2.79 (ddd, J=17.5, 13.2, 5.6 Hz, 1H), 2.72 (s, 3H), 2.52 (dd, J=4.0, 2.2 Hz, 1H), 2.13-2.03 (m, 1H), 1.99 (dddd, J=11.1, 8.3, 5.5, 2.8 Hz, 1H).
LCMS (m/z [M+H]+): 285.9
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (39% yield) using 2-(((tert-butoxycarbonyl)amino)methyl)thiazole-4-carboxylic acid (50 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.83 (s, 1H), 8.59 (d, J=8.5 Hz, 1H), 8.21 (s, 1H), 7.86 (t, J=6.1 Hz, 1H), 4.76 (ddd, J=12.6, 8.5, 5.3 Hz, 1H), 4.43 (d, J=6.1 Hz, 2H), 2.79 (ddd, J=17.3, 13.7, 5.5 Hz, 1H), 2.59-2.53 (m, 1H), 2.19 (qd, J=13.0, 4.5 Hz, 1H), 1.97 (dtd, J=12.9, 5.4, 2.6 Hz, 1H), 1.42 (s, 9H).
LCMS (m/z [M-Boc+H]+): 268.9
To a solution of 5-(methylamino)thiazole-4-carboxylic acid (1 eq.) in isopropanol and water were added 3-aminopiperidine-2,6-dione (hydrochloride salt, 3.0 eq.), N-methylmorpholine (3 eq.) and DMTMM (1.5 eq.). The reaction mixture was stirred overnight at RT, concentrated under reduced pressure and purified by HPLC to give N-(2,6-dioxopiperidin-3-yl)-5-(methylamino)thiazole-4-carboxamide (13% yield).
1H NMR (500 MHz, DMSO) δ 10.78 (s, 1H), 8.11 (d, J=0.9 Hz, 1H), 8.04 (d, J=8.3 Hz, 1H), 7.60 (q, J=5.0 Hz, 1H), 4.64 (ddd, J=12.4, 8.3, 5.3 Hz, 1H), 2.92 (d, J=5.0 Hz, 3H), 2.76 (ddd, J=17.2, 13.7, 5.5 Hz, 1H), 2.59-2.51 (m, 1H), 2.23-2.12 (m, 1H), 1.96 (dtd, J=13.0, 5.4, 2.6 Hz, 1H).
LCMS (m/z [M+H]+): 268.7
Step A: To a stirred solution of ethyl 5-bromothiazole-4-carboxylate (2.0 g, 8.475 mmol, 1 eq) in methanol (24 mL) was added NaOMe (25% in MeOH) (3.8 ml, 16.95 mmol, 2 eq). The reaction mixture was refluxed for 2 h, cooled to RT and quenched by saturated ammonium chloride solution (10 mL). The mixture was concentrated under reduced pressure and purified by flash column chromatography to give methyl 5-methoxythiazole-4-carboxylate (27% yield).
Step B: To a stirring solution of methyl 5-methoxythiazole-4-carboxylate (100 mg, 0.578 mmol, 1 eq) in a solution of THF, MeOH, H2O (4:2:1) (7 mL) was added LiOH, H2O (73 mg, 1.734 mmol, 3 eq). The reaction mixture was stirred at RT for 16 h, evaporated, redissolved in water and washed with ethyl acetate. The aqueous layer was acidified by 0.5 M HCl, extracted with 10% MeOH in DCM, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give 5-methoxythiazole-4-carboxylic acid (32% yield).
Step C: N-(2,6-dioxopiperidin-3-yl)-5-methoxythiazole-4-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (9% yield) using 5-methoxythiazole-4-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.81 (s, 1H), 8.51 (s, 1H), 8.23 (d, J=8.2 Hz, 1H), 4.67 (ddd, J=12.5, 8.2, 5.3 Hz, 1H), 4.04 (s, 3H), 2.83-2.73 (m, 1H), 2.52 (dt, J=3.9, 2.5 Hz, 1H), 2.19-2.09 (m, 1H), 1.97 (dtd, J=12.7, 5.5, 2.6 Hz, 1H).
LCMS (m/z [M+H]+): 269.8
Step A: To a stirring solution of methyl 2-amino-5-bromothiazole-4-carboxylate (1 g, 4.255 mmol, 1 eq) in methanol (30 mL) was added NaOMe (25% in MeOH) (2.3 ml, 10.638 mmol, 2.5 eq). The reaction mixture was refluxed for 1.5 h, cooled to RT and quenched by saturated ammonium chloride solution (10 mL). The mixture was concentrated under reduced pressure and purified by flash column chromatography to give methyl 2-amino-5-methoxythiazole-4-carboxylate (50% yield).
Step B: Methyl 2-amino-5-methoxythiazole-4-carboxylate (400 mg, 2.128 mmol. 1 eq.) was dissolved in DCM then were added triethylamine (0.532 mmol, 2 eq.) and Boc2O (0.532 mmol, 2 eq). The reaction mixture was stirred at RT for 18 h, diluted with DCM and washed successively with water and brine, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give methyl 2-((tert-butoxycarbonyl)amino)-5-methoxythiazole-4-carboxylate (49% yield).
Step C: To a stirring solution of methyl 2-((tert-butoxycarbonyl)amino)-5-methoxythiazole-4-carboxylate (300 mg, 1.042 mmol, 1 eq) in THF:MeOH:H2O 3:2:1 (12 mL) was added LiOH.H2O (131 mg, 3.125 mmol, 3 eq). The reaction mixture was stirred at RT for 16 h, evaporated, redissolved in water and washed with ethyl acetate. The aqueous layer was acidified by 0.5 M HCl, extracted with 10% MeOH in DCM, dried over Na2SO4, concentrated under reduced pressure and triturated with ether and pentane to give 2-((tert-butoxycarbonyl)amino)-5-methoxythiazole-4-carboxylic acid (49% yield).
Step D: Tert-butyl (4-((2,6-dioxopiperidin-3-yl)carbamoyl)-5-methoxythiazol-2-yl)carbamate was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (50% yield), and 2-((tert-butoxycarbonyl)amino)-5-methoxythiazole-4-carboxylic acid (20 mg) as a starting material.
Step E: To a solution of tert-butyl (4-((2,6-dioxopiperidin-3-yl)carbamoyl)-5-methoxythiazol-2-yl)carbamate (19.6 mg, 0.051 mmol, 1 eq.) in water (3 mL) and dioxane (3 mL) was added 36% HCl (1.5 mL). The reaction was stirred at RT for 3 h and concentrated under reduced pressure to give 2-amino-N-(2,6-dioxopiperidin-3-yl)-5-methoxythiazole-4-carboxamide hydrochloride (100% yield).
1H NMR (500 MHz, DMSO) δ 10.84 (s, 1H), 7.75 (d, J=7.8 Hz, 1H), 7.41-6.65 (m, 2H), 4.63 (ddd, J=12.0, 7.8, 5.8 Hz, 1H), 3.89 (s, 3H), 2.75 (ddd, J=17.3, 13.1, 6.2 Hz, 1H), 2.60-2.52 (m, 1H), 2.11-1.98 (m, 2H).
LCMS (m/z [M+H]+): 285.0
Step A: To a stirred solution of ethyl 5-bromothiazole-4-carboxylate (2.0 g, 8.475 mmol) in methanol (24 mL) was added NaOMe (25% in MeOH) (3.8 ml, 16.95 mmol, 2 eq). Reaction mixture was then refluxed for 2 h, cooled to RT and quenched by ammonium chloride solution. The product was extracted with ethyl acetate, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give methyl 5-methoxythiazole-4-carboxylate (27% yield).
Step B: To a solution of methyl 5-methoxythiazole-4-carboxylate (70 mg, 0.405 mmol, 1 eq) in THF (5 mL) was added N-bromosuccinimide (288 mg, 1.618 mmol, 4 eq) and the reaction mixture was stirred at RT for 24 h. The reaction mixture was diluted with ethyl acetate, washed with water and brine, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give methyl 2-bromo-5-methoxythiazole-4-carboxylate (73% yield).
Step C: To a solution of methyl 2-bromo-5-methoxythiazole-4-carboxylate (1.0 g, 3.968 mmol, 1 eq) in THF (30 mL) and water (15 mL) was added triethylamine (2.701 ml, 19.841 mmol, 5 eq) and the solution was purged with argon for 10 min. Xantphos (0.115 g, 0.198 mmol, 0.05 eq) and Pd(OAc)2 (44 mg, 0.198 mmol, 0.05 eq) were added and the reaction mixture was stirred at 60° C. under CO (50 psi) for 16 h. The reaction mixture was cooled to RT, diluted with water and washed with ethyl acetate. The aqueous layer was acidified by 2M HCl solution, extracted with 15% MeOH in DCM, dried over Na2SO4 and concentrated under reduced pressure to give 5-methoxy-4-(methoxycarbonyl)thiazole-2-carboxylic acid (30% yield).
Step D: To a solution of 5-methoxy-4-(methoxycarbonyl)thiazole-2-carboxylic acid (300 mg, 1.382 mmol, 1 eq) in tert-butanol (15 mL) was added 2-tert-butyl-1,3-diisopropylisourea (829 mg, 4.147 mmol, 3 eq) and the reaction mixture was stirred at RT for 16 h, diluted with ethyl acetate and washed by water, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give 2-tert-butyl 4-methyl 5-methoxythiazole-2,4-dicarboxylate (31% yield).
Step E: To a solution of 2-tert-butyl 4-methyl 5-methoxythiazole-2,4-dicarboxylate (220 mg, 0.806 mmol, 1 eq) in DCE (5 mL) was added trimethyltin hydroxide (728 mg, 4.029 mmol, 5 eq). Reaction mixture was stirred at 90° C. for 6 h, filtered, the filtrate was concentrated under reduced pressure and purified by HPLC to give 2-(tert-butoxycarbonyl)-5-methoxythiazole-4-carboxylic acid (11% yield).
Step F: tert-butyl 4-((2,6-dioxopiperidin-3-yl)carbamoyl)-5-methoxythiazole-2-carboxylate was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (39% yield) using 2-(tert-butoxycarbonyl)-5-methoxythiazole-4-carboxylic acid (16.5 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.83 (s, 1H), 8.24 (d, J=8.2 Hz, 1H), 4.77-4.65 (m, 1H), 4.14 (s, 3H), 2.77 (ddd, J=17.2, 13.8, 5.5 Hz, 1H), 2.60-2.53 (m, OH), 2.17 (qd, J=12.9, 4.5 Hz, 1H), 1.96 (dtd, J=12.9, 5.5, 2.6 Hz, 1H), 1.56 (s, 9H).
Step G: tert-butyl 4-((2,6-dioxopiperidin-3-yl)carbamoyl)-5-methoxythiazole-2-carboxylate (6 mg, 0.016 mmol) was dissolved in DCM (0.5 mL) and trifluoroacetic acid (0.092 mL) was added. The Reaction was stirred at RT for 2 h and concentrated under reduced pressure to give 4-((2,6-dioxopiperidin-3-yl)carbamoyl)-5-methoxythiazole-2-carboxylic acid (71% yield).
1H NMR (500 MHz, DMSO) δ 14.03 (s, 1H), 10.82 (s, 1H), 8.27 (d, J=8.3 Hz, 1H), 4.71 (ddd, J=12.5, 8.3, 5.4 Hz, 1H), 4.14 (s, 3H), 2.77 (ddd, J=17.1, 13.7, 5.5 Hz, 1H), 2.58 (s, 1H), 2.16 (qd, J=12.9, 4.5 Hz, 1H), 1.96 (dtd, J=12.9, 5.4, 2.6 Hz, 1H).
LCMS (m/z [M+H]+): 314.0
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (67% yield) using 5-cyclopropyl-1,3-thiazole-4-carboxylic (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.83 (s, 1H), 8.86 (d, J=0.7 Hz, 1H), 8.59 (d, J=8.3 Hz, 1H), 4.73 (ddd, J=12.6, 8.3, 5.3 Hz, 1H), 3.35-3.31 (m, 1H), 2.78 (ddd, J=17.4, 13.7, 5.6 Hz, 1H), 2.59-2.51 (m, 1H), 2.25-2.15 (m, 1H), 2.03-1.97 (m, 1H), 1.28-1.20 (m, 2H), 0.68 (pd, J=4.4, 1.9 Hz, 2H).
LCMS (m/z [M+H]+): 280.1
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (4% yield), and 4,5-dimethyl-2-pivalamidothiophene-3-carboxylic acid (22.5 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 11.17 (s, 1H), 10.91 (s, 1H), 7.93 (d, J=8.1 Hz, 1H), 4.80 (ddd, J=13.0, 8.1, 5.4 Hz, 1H), 2.81 (ddd, J=18.4, 13.5, 5.6 Hz, 1H), 2.59-2.51 (m, 1H), 2.24 (s, 3H), 2.22 (s, 3H), 2.14 (qd, J=13.0, 4.5 Hz, 1H), 2.07-2.01 (m, 1H), 1.21 (s, 9H).
LCMS (m/z [M+H]+): 366.15
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (27% yield) using 2-benzamido-4,5-dimethylthiophene-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 11.77 (s, 1H), 10.94 (s, 1H), 8.11 (d, J=8.2 Hz, 1H), 7.90 (dt, J=7.1, 1.4 Hz, 2H), 7.68-7.61 (m, 1H), 7.58 (dd, J=8.4, 6.9 Hz, 2H), 4.90-4.80 (m, 1H), 2.82 (ddd, J=18.5, 13.3, 5.7 Hz, 1H), 2.58 (d, J=3.5 Hz, 1H), 2.29 (s, 3H), 2.24 (s, 3H), 2.21-2.04 (m, 2H).
LCMS (m/z [M−1-1]1: 386.0
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (3.9% yield) 2-(4-methoxybenzamido)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid (30 mg) as a starting material.
1H NMR (500 MHz, DMSO, 300 K): δ 12.28 (s, 1H), 10.92 (s, 1H), 7.85 (d, J=8.8 Hz, 2H), 7.72 (d, J=8.0 Hz, 1H), 7.14 (d, J=8.6 Hz, 2H), 4.91-4.80 (m, 1H), 3.85 (s, 3H), 2.87-2.79 (m, 1H), 2.77 (d, J=5.5 Hz, 2H), 2.67 (s, 2H), 2.59-2.53 (m, 1H), 2.24-2.02 (m, 2H), 1.77 (s, 4H).
LCMS (m/z [M+H]+): 441.9
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (25% yield) using triethylamine instead of DIPEA and 2-(2-morpholinoacetamido)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid (18 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 11.84 (s, 1H), 10.86 (s, 1H), 7.69 (d, J=8.2 Hz, 1H), 4.77 (ddd, 0.1=13.0, 8.1, 5.4 Hz, 1H), 3.71-3.59 (m, 4H), 3.19 (s, 2H), 2.83 (ddd, J=17.4, 13.6, 5.7 Hz, 1H), 2.78-2.67 (m, 2H), 2.67-2.61 (m, 2H), 2.58-2.53 (m, 1H), 2.48 (d, J=4.2 Hz, 2H), 2.14 (qd, J=12.9, 4.4 Hz, 1H), 2.03 (ddt, J=13.6, 6.8, 3.3 Hz, 1H), 1.82-1.67 (m, 4H).
LCMS (m/z [M+H]+): 435.1
Step A: To a solution of methyl 2,4-dimethylthieno[3,4-b]pyridine-7-carboxylate (25.0 mg, 0.113 mmol, 1.000 eq) in a mixture of H2O (1.0 mL), THF (1.0 mL) and MeOH (1.0 mL) was added 1M LiOH (2.0 mL, 2.000 mmol, 17.7 eq) and the reaction was stirred at RT for 24 h and neutralized with 1M HCl. After concentration under reduced pressure 2,4-dimethylthieno[3,4-b]pyridine-7-carboxylic acid was used in the next step without further purification.
Step B: N-(2,6-dioxopiperidin-3-yl)-2,4-dimethylthieno[3,4-b]pyridine-7-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (53% yield), and 2,4-dimethylthieno[3,4-b]pyridine-7-carboxylic acid (23.4 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.95 (s, 1H), 10.08 (d, J=6.8 Hz, 1H), 8.52 (s, 1H), 7.02 (t, J=1.1 Hz, 1H), 4.84 (ddd, J=12.3, 6.8, 5.3 Hz, 1H), 2.82 (ddd, J=17.4, 13.6, 5.5 Hz, 1H), 2.61 (s, 3H), 2.59-2.53 (m, 4H), 2.32 (dtd, J=12.9, 5.4, 2.4 Hz, 1H), 2.12 (qd, J=12.9, 4.4 Hz, 1H).
LCMS (m/z [M+H]+): 317.9
4-chloro-2-(trifluoromethyl)thieno[3,4-b]pyridine-7-carboxylic acid (1 eq.) and DMF (1 μL) were dissolved in DCM (3 mL) and then oxalyl chloride (0.018 mL, 0.213 mmol, 3.000 eq) was added. The reaction mixture was stirred at RT for 2 h and concentrated under reduced pressure. The material was dissolved in DMF (2 mL), 3-aminopiperidine-2,6-dione hydrochloride (2 eq.) and DIPEA (2 eq.) were added and the reaction mixture was stirred at RT for 72 h and purified by preparative HPLC to give 4-chloro-N-(2,6-dioxopiperidin-3-yl)-2-(trifluoromethyl)thieno[3,4-b]pyridine-7-carboxamide (6% yield).
1H NMR (500 MHz, DMSO): δ 11.03 (s, 1H), 9.30 (d, J=6.4 Hz, 1H), 8.90 (s, 1H), 8.03 (s, 1H), 4.93-4.84 (m, 1H), 2.82 (ddd, J=18.8, 13.6, 5.5 Hz, 1H), 2.61-2.54 (m, 1H), 2.40 (ddd, J=7.2, 5.3, 2.7 Hz, 1H), 2.11-2.02 (m, 1H).
LCMS (m/z [M+H]+): 391.8
Step A: To a solution of methyl 4-hydroxy-2-(trifluoromethyl)thieno[3,4-b]pyridine-7-carboxylate (30.0 mg, 0.108 mmol, 1.000 eq) in MeOH (2.0 mL) was added NaOH (216 mg, 5.411 mmol, 50 eq). The reaction was stirred at RT for 24 h and neutralized with 1M HCl. After concentration under reduced pressure 4-hydroxy-2-(trifluoromethyl)thieno[3,4-b]pyridine-7-carboxylic acid was used in the next step without further purification.
Step B: N-(2,6-dioxopiperidin-3-yl)-4-hydroxy-2-(trifluoromethyl)thieno[3,4-b]pyridine-7-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (24% yield), and 4-hydroxy-2-(trifluoromethyl)thieno[3,4-b]pyridine-7-carboxylic acid (28 mg) as a starting material.
1H NMR (500 MHz, DMSO): δ 12.98 (s, 1H), 10.98 (s, 1H), 9.53 (d, J=6.6 Hz, 1H), 8.76 (s, 1H), 6.76 (s, 1H), 4.91-4.81 (m, 1H), 2.81 (ddd, J=18.6, 13.4, 5.4 Hz, 1H), 2.60-2.54 (m, 1H), 2.38-2.33 (m, 1H), 2.08-1.99 (m, 1H).
LCMS (m/z [M+H]+): 374.0
N-bromosuccinimide (96.8 mg, 0.544 mmol, 1.1 eq) was added to a suspension of N-(2,6-dioxopiperidin-3-yl)thieno[3,4-b]pyridine-7-carboxamide (143.0 mg, 0.494 mmol, 1.000 eq,) in DMF (4.9 mL) at ambient temperature. The reaction mixture was heated to 60° C. and stirred for 3 h. The obtained crude compound was purified by HPLC to give 5-bromo-N-(2,6-dioxopiperidin-3-yl)thieno[3,4-b]pyridine-7-carboxamide (15% yield).
1H NMR (500 MHz, DMSO) δ 10.97 (s, 1H), 9.53 (d, J=7.4 Hz, 1H), 8.87 (dd, J=4.0, 1.5 Hz, 1H), 8.11 (dd, J=8.9, 1.5 Hz, 1H), 7.41 (dd, J=8.9, 4.0 Hz, 1H), 4.91 (ddd, J=12.8, 7.3, 5.6 Hz, 1H), 2.84 (ddd, J=17.5, 13.4, 5.7 Hz, 1H), 2.57 (ddd, J=17.4, 4.3, 2.3 Hz, 1H), 2.30-2.13 (m, 2H)
LCMS (m/z [M+H]+): 368.37
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (15% yield) using 4-chloro-1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 12.40 (s, 1H), 10.82 (s, 1H), 8.38 (d, J=8.3 Hz, 1H), 8.22 (d, J=5.1 Hz, 1H), 7.96 (s, 1H), 7.25 (d, J=5.1 Hz, 1H), 4.79-4.72 (m, 1H), 2.79 (ddd, J=17.9, 9.7, 7.0 Hz, 1H), 2.59-2.52 (m, 1H), 2.12-2.01 (m, 2H).
LCMS (m/z [M+H]+): 306.9
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (61% yield) using 1-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.84 (s, 1H), 8.44 (dd, J=7.9, 1.6 Hz, 1H), 8.34 (dd, J=4.7, 1.6 Hz, 1H), 8.30 (d, J=8.3 Hz, 1H), 8.21 (s, 1H), 7.23 (dd, J=7.9, 4.7 Hz, 1H), 4.78 (ddd, J=12.2, 8.3, 5.3 Hz, 1H), 3.88 (s, 3H), 2.80 (ddd, I=17.3, 13.1, 5.5 Hz, 1H), 2.59-2.51 (m, 1H), 2.12 (qd, J=12.8, 4.5 Hz, 1H), 2.00 (dtd, J=12.8, 5.5, 3.0 Hz, 1H).
LCMS (m/z [M+H]+): 286.9
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (31% yield) using 5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 12.40 (s, 1H), 10.85 (s, 1H), 8.44 (d, J=2.5 Hz, 1H), 8.39 (d, J=8.4 Hz, 1H), 8.30 (d, J=2.4 Hz, 1H), 8.27 (d, I=2.9 Hz, 1H), 4.79 (ddd, J=12.2, 8.3, 5.3 Hz, 1H), 2.81 (ddd, J=17.3, 13.2, 5.5 Hz, 1H), 2.59-2.52 (m, 1H), 2.11 (qd, J=12.8, 4.4 Hz, 1H), 2.01 (dtd, J=13.0, 5.4, 2.9 Hz, 1H).
LCMS (m/z [M+H]+): 307.2
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (8% yield), and 4-chloro-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylic acid (20.0 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.82 (s, 1H), 8.55 (s, 1H), 8.49 (s, 1H), 8.36 (s, 1H), 8.06 (s, 1H), 4.76 (q, J=8.5 Hz, 1H), 2.78 (ddd, J=18.0, 10.3, 8.4 Hz, 1H), 2.56 (dt, 1=17.2, 3.8 Hz, 1H), 2.06 (h, J=5.0, 4.4 Hz, 2H).
LCMS (m/z [M+H]+): 308.0
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (40% yield) using 1H-pyrazolo[4,3-b]pyridine-3-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO): δ 14.01 (s, 1H), 10.93 (s, 1H), 9.07 (d, J=7.5 Hz, 1H), 8.69 (dd, J=4.4, 1.3 Hz, 1H), 8.19 (dd, J=8.5, 1.0 Hz, 1H), 7.51 (dd, J=8.5, 4.4 Hz, 1H), 4.92 (ddd, J=7.5, 6.7, 4.2 Hz, 1H), 2.90-2.78 (m, 1H), 2.60-2.54 (m, 1H), 2.25-2.11 (m, 2H).
LCMS (m/z [M+H]+): 273.8
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions B, above (19% yield), with 2-acetamido-4,5,6,7-tetrahydro-1-benzothiophene-3-carboxylic acid as a starting material.
1H NMR (500 MHz, DMSO) δ 10.99 (s, 1H), 10.82 (s, 1H), 7.99 (d, J=8.4 Hz, 1H), 4.91-4.76 (m, 1H), 2.82 (ddd, J=17.5, 13.1, 6.1 Hz, 1H), 2.73-2.54 (m, 5H), 2.16 (s, 3H), 2.15-2.00 (m, 2H), 1.84-1.67 (m, 4H).
LCMS (m/z [M−H]−): 347.8
N-chlorosuccinimide (0.059 g, 0.442 mmol, 1.1 eq) was added to a suspension of N-(2,6-dioxopiperidin-3-yl)thieno[3,4-b]pyridine-7-carboxamide (0.116 g, 0.401 mmol) in DMF (5 mL) at RT. The reaction mixture was heated to 60′C and stirred for 3 h. The obtained crude compound was purified by HPLC to give 5-chloro-N-(2,6-dioxopiperidin-3-yl)thieno[3,4-b]pyridine-7-carboxamide (43% yield).
1H NMR (400 MHz, DMSO) δ 10.97 (s, 1H), 9.51 (d, J=7.4 Hz, 1H), 8.91-8.83 (m, 1H), 8.19 (d, J=8.8 Hz, 1H), 7.41-7.33 (m, 1H), 4.95-4.84 (m, 1H), 2.89-2.74 (m, 1H), 2.62-2.55 (m, 1H), 2.28-2.11 (m, 2H)
LCMS (m/z [M+H]+): 323.8
Step A: To an ice-cold solution of 2-bromo-3-(bromomethyl)pyridine 2 (10.5 g, 42.0 mmol) in THF (100 mL) was added methyl thioglycolate (4.089 g, 18.124 mmol) followed by Et3N under stirring. The mixture was warmed to RT and stirred for further 30 min. The reaction mixture was diluted with water and extracted with DCM, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give methyl 2-{[(2-bromopyridin-3-yl)methyl]sulfanyl}acetate (53% yield).
Step B: A solution of methyl 2-{[(2-bromopyridin-3-yl)methyl]sulfanyl}acetate (4.5 g, 16.295 mmol) in THF (25 mL) was added slowly to a suspension of KH (1.307 g, 32.591 mmol) and stirred for 20 min at room temperature. The reaction mixture was then cooled to −78° C. and treated with saturated aqueous NH4Cl solution, warmed to RT, extracted with DCM, dried over Na2SO4, concentrated under reduced pressure and purified by flash column chromatography to give methyl 5H,7H-thieno[3,4-b]pyridine-7-carboxylate (56% yield).
Step C: To the stirred solution of methyl 5H,7H-thieno[3,4-b]pyridine-7-carboxylate (3 g, 15.385 mmol) in CHCl3 (25 mL) was added activated MnO2 (13.375 g, 153.846 mmol) and the reaction mixture was stirred at RT for 16 h, filtered through celite bed, concentrated under reduced pressure and purified by flash column chromatography to give methyl thieno[3,4-b]pyridine-7-carboxylate (46% yield).
Step D: To a stirred solution of methyl thieno[3,4-b]pyridine-7-carboxylate (1.5 g, 7.772 mmol) in THF:MeOH:H2O, 4:2:1 (14 mL) was added LiOH.H2O (1.304 g, 31.088 mmol) at 0° C. and then ice-bath was removed and the mixture was stirred at RT for 2.5 h. Saturated aqueous citric acid solution was added and the product was extracted with 10% MeOH in DCM, dried over Na2SO4, concentrated under reduced pressure and purified by HPLC to give thieno[3,4-b]pyridine-7-carboxylic acid (72 mg, 5%).
Step E: Synthesis of N-(2,6-dioxopiperidin-3-yl)thieno[3,4-b]pyridine-7-carboxamide was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (69% yield), and thieno[3,4-b]pyridine-7-carboxylic acid (25.0 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.95 (s, 1H), 9.71 (d, J=7.4 Hz, 1H), 8.83 (dd, J=4.0, 1.6 Hz, 1H), 8.59 (s, 1H), 8.32 (dd, J=8.8, 1.6 Hz, 1H), 7.29 (dd, J=8.7, 4.0 Hz, 1H), 4.92 (ddd, J=12.7, 7.4, 5.4 Hz, 1H), 2.89-2.79 (m, 1H), 2.60-2.52 (m, 1H), 2.30-2.11 (m, 2H).
LCMS (m/z [M+H]+): 290.0
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above, (50% yield) using 3-methoxythiophene-2-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.87 (s, 1H), 7.89 (d, J=7.4 Hz, 1H), 7.75 (d, J=5.5 Hz, 1H), 7.14 (d, J=5.5 Hz, 1H), 4.78-4.63 (m, 1H), 3.98 (s, 3H), 3.31 (s, 3H), 2.77 (ddd, J=17.4, 13.1, 6.3 Hz, 1H), 2.54-2.51 (m, 1H), 2.17-2.04 (m, 2H).
LCMS (m/z [M+H]+): 268.9
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions D, above, (42% yield) using 4-methoxythiophene-3-carboxylic acid (20 mg) and (S)-3-aminopiperidine-2,6-dione as a starting material.
1H NMR (500 MHz, DMSO) δ 10.89 (s, 1H), 8.12 (d, J=7.4 Hz, 1H), 8.09 (d, J=3.6 Hz, 1H), 6.81 (d, J=3.6 Hz, 1H), 4.78-4.65 (m, 1H), 3.89 (s, 3H), 2.84-2.72 (m, 1H), 2.56-2.52 (m, 1H), 2.15-2.06 (m, 2H).
LCMS (m/z [M+H]+): 268.9
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions D, above, (40% yield) using 4-methoxythiophene-3-carboxylic acid (20 mg) and (R)-3-aminopiperidine-2,6-dione as a starting material.
1H NMR (500 MHz, DMSO) δ 10.89 (s, 1H), 8.12 (d, J=7.4 Hz, 1H), 8.09 (d, J=3.6 Hz, 1H), 6.81 (d, J=3.6 Hz, 1H), 4.76-4.68 (m, 1H), 3.89 (s, 3H), 2.84-2.72 (m, 1H), 2.55-2.52 (m, 1H), 2.14-2.06 (m, 2H).
LCMS (m/z [M+H]+): 268.8
This compound was synthesized using the general procedure shown in Reaction Scheme 1 and Synthetic Conditions C, above (79% yield) using 2-(methoxymethyl)thiazole-5-carboxylic acid (20 mg) as a starting material.
1H NMR (500 MHz, DMSO) δ 10.89 (s, 1H), 8.95 (d, J=8.3 Hz, 1H), 8.34 (s, 1H), 4.77-4.70 (m, 3H), 4.72 (s, 2H), 3.40 (d, J=17.0 Hz, 3H), 2.79 (ddd, J=17.5, 13.3, 5.6 Hz, 1H), 2.52 (dd, J=5.7, 3.8 Hz, 1H), 2.10 (qd, J=13.0, 4.5 Hz, 1H), 1.99 (dddd, J=11.0, 8.3, 5.5, 2.8 Hz, 1H).
LCMS (m/z [M+H]+): 283.9
To the suspension of tert-butyl (5-((2,6-dioxopiperidin-3-yl)carbamoyl)thiazol-2-yl)carbamate (71, 30 mg, 0.085 mmol, 1 eq) in DCM (1.5 mL) was added TFA (0.2 mL) and mixture was stirred for 18 h at RT, concentrated under reduced pressure and purified by HPLC to give 2-amino-N-(2,6-dioxopiperidin-3-yl)thiazole-5-carboxamide (yield 37%).
1H NMR (500 MHz, DMSO) δ 10.81 (s, 1H), 8.35 (d, J=8.4 Hz, 1H), 7.64 (s, 1H), 7.49 (s, 2H), 4.64 (ddd, J=12.5, 8.4, 5.4 Hz, 1H), 2.76 (ddd, J=17.4, 13.3, 5.6 Hz, 1H), 2.53-2.51 (m, 2H), 2.05 (qd, J=12.8, 4.3 Hz, 1H), 1.93 (dddd, J=10.7, 8.1, 5.3, 2.9 Hz, 1H).
LCMS (m/z [M+H]+): 255.2
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).
Fluorescence Polarization (FP) Assay—Results
Compounds are categorized based on their affinity to CRBN defined as Ki. As reported in Table 2 below, the compounds of the present invention interact with CRBN-DDB1 protein within similar affinity range as reported for reference compounds.
The effect of various compounds of the invention and various reference compounds on CK1α 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 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: 54, 109, POMALIDOMIDE, CC-122, and LENALIDOMIDE at the concentrations 1, 10 and 20 μM. In addition, compounds 22, 21, 108 and 110 were tested at 20 μM. The treatment with all compounds was carried out for 24 h.
Densitometry values are normalized to the loading control ((3-ACTIN) and presented as % of DMSO control, using the following labels:
≤25% for 0-25% of CK1α protein reduction,
>25% for 26-74% of CK1α protein reduction,
≥75% for 75-100% of CK1α protein reduction.
The representative results for compounds: 54, 109, POMALIDOMIDE, CC-122, and LENALIDOMIDE are shown 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 6 or 24 h incubation (3TC, 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 compounds 109, 20, 108, 111, POM, CC-122, LEN, 106, 107 at the concentrations 1, 10 and 20 μM. The remaining compounds, listed in Table 6, were tested at 20 μM. The treatment with all compounds was carried out for 24 h. Densitometry values are normalized to the loading control (β-ACTIN) and presented as % of DMSO control, using the following labels:
≤25% for 0-25% of IKZF1 protein reduction,
>25% for 26-74% of IKZF1 protein reduction,
≥75% for 75-100% of IKZF1 protein reduction.
The representative results for compounds 109, 20, 108, 111, POM, CC-122, LEN, 106, 107 are shown 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, 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 (3TC, 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: compounds 109, 20, 108, 111, POM, CC-122, LEN, 106, 107 at the concentrations 1, 10 and 20 μM. The treatment with all compounds was carried out for 24 h.
Densitometry values are normalized to the loading control ((3-ACTIN) and presented as % of DMSO control, using the following labels:
≤25% for 0-25% of IKZF3 protein reduction,
>25% for 26-74% of IKZF3 protein reduction,
≥75% for 75-100% of IKZF3 protein reduction.
The representative results for compounds 109, 20, 108, 111, POM, CC-122, LEN, 106, 107 are shown in
In summary, the compounds of invention are capable of potent degradation of CK1α, a disease relevant protein kinase. By contrast, presented neosubstrates IKZF1, IKZF3 degradation tests results for the compounds of the present invention show no to low degradation of the proteins by the compounds, as opposed to the known CK1α degraders, Lenalidomide and Pomalidomide. This innovative profile renders the compounds of the present invention useful as more selective CK1α degraders.
A list of the abbreviations used in the present application is shown in Table 8, 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.
1. A compound of Formula (Ia) or (Ib):
3. The compound of embodiment 1, having the structure:
4. The compound of any preceding embodiment, wherein T is C═O.
5. The compound of any one of embodiments 1-3, wherein T is SO2.
6. The compound of any preceding embodiment, wherein Z is NR2.
7. The compound of any preceding embodiment, wherein R2 is alkyl, benzyl, or —N[C(O)R″]2.
8. The compound of any one of embodiments 1-5, wherein Z is S.
9. The compound of any one of embodiments 1-8, wherein L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, —OR″, —NR″2, or —S(O)2R″; optionally wherein L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, or benzyl.
10. The compound of embodiment 9, wherein L is hydrogen.
11. The compound of any preceding embodiment, wherein the compound is of Formula (Ia), wherein one of Y1, Y2 and Y3 is N, and the remaining two of Y1, Y2 and Y3 are each CR.
12. The compound of embodiment 11, wherein Y1 is N, and Y2 and Y3 are CR.
13. The compound of embodiment 11, wherein Y2 is N, and Y1 and Y3 are CR; and Z is S.
14. The compound of embodiment 11, wherein Y3 is N; Y1 and Y2 are CR; and Z is S.
15. The compound of any one of embodiments 1-10, wherein the compound is of Formula (Ia), wherein one of Y1, Y2 and Y3 is CR, the remaining two of Y1, Y2 and Y3 are each N; and Z is S.
16. The compound of embodiment 15, wherein Y1 is CR; Y2 and Y3 are N.
17. The compound of embodiment 15, wherein Y2 is CR; Y1 and Y3 are N.
18. The compound of embodiment 15, wherein Y3 is CR, and Y1 and Y2 are N.
19. The compound of any one of embodiments 1-10, wherein the compound is of Formula (Ia) and Y2 and Y3 are each CR.
20. The compound of embodiment 19, wherein
39. The compound embodiment 37, having the structure:
40. The compound of any one of embodiments 37-39, wherein Z is O.
41. The compound of any one of embodiments 37-39, wherein Z is S.
42. The compound of any one of embodiments 37-39, wherein Z is NR2.
43. The compound of any one of embodiments 37-42, wherein T is C═O.
44. The compound of any one of embodiments 37-42, wherein T is SO2.
45. The compound of any one of embodiments 37-44, wherein L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, benzyl, —OR″, —NR″2, or —S(O)2R″; optionally wherein L is hydrogen, alkyl, alkenyl, aryl, heteroaryl, or benzyl.
46. The compound of embodiment 45, wherein L is hydrogen.
47. The compound of any one of embodiments 37-46, wherein Y3 is N.
48. The compound of any one of embodiments 37-46, wherein Y3 is CR.
49. The compound of any one of embodiments 37-46, wherein Y4 is N.
50. The compound of any one of embodiments 37-46, wherein Y4 is CR.
51. The compound of any one of embodiments 37-50, wherein each is a double bond or wherein each
is a single bond.
52. The compound of embodiment 51, wherein each is a double bond.
53. The compound of embodiment 52, wherein one of W1, W2, W3 and W4 is N, and the remaining three of W1, W2, W3 and W4 are each CR′.
54. The compound of embodiment 52, wherein two of W1, W2, W3 and W4 is N, and the remaining two of W1, W2, W3 and W4 are each CR′.
55. The compound of embodiment 52, wherein one of W1, W2, W3 and W4 is CR′, and the remaining three of W1, W2, W3 and W4 are each N.
56. The compound of embodiment 51, wherein each is a single bond.
57. The compound of any one of embodiments 37-56, wherein each R is independently hydrogen, halogen or —NR″C(O)R″.
58. The compound of any one of embodiments 37-57, wherein each R′ is hydrogen.
59. The compound of any preceding embodiment, wherein X1 and X2 are O.
60. The compound of any one of embodiments 1-58 wherein X1 is O and X2 is S.
61. The compound of any one of embodiments 1-58, wherein X1 is S and X2 is O.
62. The compound of any one of embodiments 1-58, wherein X1 and X2 are S.
63. The compound of any preceding embodiment, wherein n is 0.
64. The compound of any one of embodiments 1-62, wherein n is 1.
65. The compound of any one of embodiments 1-62, wherein n is 2.
66. A compound of any one of the preceding embodiments, for use as a cereblon binder.
67. A pharmaceutical composition comprising a compound of any one of embodiments 1-65.
68. A compound of any one of embodiments 1-65, or a composition according to embodiment 67, for use in medicine.
69. A compound of any one of embodiments 1-65, or a composition according to embodiment 67, for use in immune-oncology.
70. A compound of any one of embodiments 1-65, or a composition according to embodiment 67, 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.
71. 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;
| Number | Date | Country | Kind |
|---|---|---|---|
| PCT/PL2019/000108 | Nov 2019 | PL | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2020/083597 | 11/27/2020 | WO |
