Patent Grant
11013745
The present invention relates to pharmaceutical chemistry and pharmaceutical therapeutics, and generally relates to a triazolopyrimidine, triazolopyridine compounds, and pharmaceutical compositions, and their use in the treatment of tumor diseases. In particular, such compounds can be prepared for medicine of the treatment of PRC2-mediated diseases or conditions.
Polycomb Repressive Complex 2 (PRC2) is a core member of the Polycomb Group. It has histone methyltransferase activity, and can specifically catalyze the trimethylation modification (H3K27me3) of lysine at position 27 of histone H3 to suppress the expression of specific genes. The methyltransferase activity of PRC2 is derived from its catalytic member EZH2. However, EZH2 has no catalytic activity when it is alone. It needs to form a complex with at least two other members of PRC2, i.e., EED and SUZ12, to catalyze the methylation modification. Therefore, EZH2, EED and SUZ12 are considered as the core components of the PRC2 complex. Recent studies have found that the core components of PRC2 are overexpressed in a variety of tumor cells, and their abnormal activity is the direct reason of the onset and deterioration of various malignant tumors. At the same time, recent gene sequencing results of lymphoma patients showed that EZH2 exhibits activating mutation in patients with germinal center B cell lymphoma (GCB-DLBCL). The mutated EZH2 alters the substrate specificity of PRC2, thereby increases the level of H3K27me3 in cells. Down-regulating the expression of EZH2 or other core components by the siRNA method will significantly inhibit the proliferation of lymphoma cells, which indicates that the occurrence and development of GCB-DLBCL is closely related to the excessive activation of PRC2. Therefore, PRC2 is a very promising target for the development of anticancer drugs, and the discovery of inhibitors that target PRC2 is currently a hot topic in the pharmaceutical industry. Recently, two major pharmaceutical companies, Novartis and AbbVie, have invented a small molecules that inhibit PRC2 activity by targeting EED (Reference: Novartis EED226, US 2016/0176682, J. Med. Chem. 2017, 60, 2215-2226, J. Med. Chem. 2017, 60, 415-427, Nat. Chem. Biol. 2017, 13, 381-388; AbbVie's A-395, Nat. Chem. Biol. 2017, 13, 389-395), this kind of compound shows very strong inhibitory activity at the molecular level, cell level and animal experiments. In summary, the PRC2 complex is considered to be a key driver for the occurrence and development of a variety of malignant tumors, and the development of inhibitors that target EED to inhibit PRC2 activity is currently very hot in the industry and is conducive to the use in the related new drug development.
The present invention relates to a triazolopyrimidine, triazolopyndine compounds as shown in Formula I, and pharmaceutical compositions thereof, by combining the evaluation of binding activity and related biological experiments, it can be used for the preparation for the drugs for the treatment of EED and/or PRC2-mediated diseases or conditions.
It is an object of the present invention to provide a triazolopyrimidine, triazolopyridine compounds, pharmaceutically acceptable salts, enantiomers, diastereomers or racemates.
It is another object of the present invention to provide a method for preparing the compound.
It is a further object of the present invention to provide a pharmaceutical composition comprising a therapeutically effective amount of one or more of the above compounds or a pharmaceutically acceptable salt thereof.
It is yet another object of the present invention to provide the use of the above compound in the preparation of a medicament for treating a disease or condition mediated by EED and/or PRC2.
It is yet another object of the present invention to provide a method for treating a disease or condition mediated by EED and/or PRC2, characterized in that a therapeutically effective amount of one or more of the above-mentioned compounds or a pharmacological salt thereof is administered to a subject.
Specifically, according to one aspect of the invention, it provides a compound of formula I:
wherein
1) X1 is independently selected from N and C—CN;
2) R2 is independently selected from H, halogen, C1-C4 haloalkyl and C1-C4 alkyl;
3) A is independently selected from the following structures:
is a single bond or double bond;
R3, R4 and R5 are independently selected from H, halogen, C1-C4 alkyl, C1-C4 haloalkyl, —O—(C1-C4 alkyl), C1-C4 haloalkoxy, C3-C6 cycloalkyl;
R6 is independently selected from H, OH, ═O and C1-C4 alkyl;
R7 is independently selected from H, OH, halogen, CN and C1-C4 alkyl;
n is each independently selected from 0, 1 and 2;
X2 is independently selected from O, NRa and S(O)p heteroatoms;
Each Ra is independently selected from H, O, C1-C10 alkyl substituted by 0-2 Rb, C1-C6 haloalkyl, —O—(C1-C6 alkyl), C1-C6 haloalkoxy, C3-C6 cycloalkyl, —C(═O)(C1-C4 alkyl), —CO2 (C1-C4 alkyl), heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p, —C(═O)H, aryl, 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S;
Rb is independently selected from halogen, OH, NH2, NHC(═O)(C1-C4 alkyl), NHS(═O)2 (C1-C4 alkyl), ═O, CN, C1-C4 alkyl and C1-C4 alkoxy;
p is each independently selected from 0, 1 and 2;
4) R1 is independently selected from the following structures:
is a single bond or double bond;
4a) R1A is independently selected from H, hydroxy, halogen, CN, —(O)z—(C1-C10 alkyl comprising 0-2 of substituent Rc), C1-C6 alkyl group, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, SCF3, C3-C8 cycloalkyl, —C(═O)(C1-C4 alkyl), —C(═O)NH(C1-C4 alkyl), amino, C1-C6 linear, branched and cyclic alkylamino, heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p, —C(═O)H, aryl, 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S; wherein the aryl and heteroaryl may be substituted by 0-2 of R1X;
p is each independently selected from 0, 1 and 2;
Rc is independently selected from OH, halogen, CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C8 cycloalkyl, —(OCH2CH2)mORd, NHC(═O)NRdRe, NHC(═S)NRdRe, —NHC(═NH)NRdRe, (OCH2CH2)mNRdRe, —C(═O)Rd, —S(═O)Rd, —C(═O)NRdRe, —S(═O)2Rd, —NHC(═O)Rd, —NHC(═S)Rd, —NHS(═O)2Rd, —S(═O)2NHRd, heteroalkyl and heterocycloalkyl comprising carbon atoms and 1 to 2 heteroatoms selected from N, NRa, O and S(O)p, aryl, and heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, NRa, O and S(O)p, wherein the aryl and heteroaryl may be substituted by 0-2 of R1X;
Rd and Re are independently selected from H, C1-C6 alkyl comprising 0-2 of Rb, C1-C6 haloalkyl, C3-C6 cycloalkyl, —C(═O)(C1-C4 alkyl). —CO2(C1-C4 alkyl), —C(═O)NH(C1-C4 alkyl), C1-C6 branched or cyclic heteroalkyl comprising 0-2 of heteroatoms selected from O, N, and S(O)p, —C(═O)H, aryl and heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, NRa, O and S(O)p, wherein the aryl and heteroaryl may be substituted by 0-2 of R1X;
Each Ra is independently selected from H, O, C1-C10 alkyl substituted by 0-2 of Rb, C1-C6 haloalkyl, —O—(C1-C6 alkyl), C1-C6 haloalkoxy, C3-C6 cycloalkyl, —C(═O)(C1-C4 alkyl). —CO2(C1-C4 alkyl), heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p, —C(═O)H, aryl, 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S;
Rb is independently selected from halogen, OH, NH2, NHC(═O)(C1-C4 alkyl), NHS(═O)2(C1-C4 alkyl), ═O, CN, C1-C4 alkyl and C1-C4 alkoxy;
p is each independently selected from 0.1 and 2;
Rd and Re can be connected in the form of
wherein the Z1 may be selected from C1-C6 alkyl comprising 0-2 of substituent Rb, C1-C6 heteroalkyl comprising 0-2 of substituent heteroatoms of O, N, S(O)p, O, —N(C1-C6 alkyl), —NH, —N(C═O) C1-C6 alkyl, —NS(═O)2(C1-C6 alkyl), S(O)p; Rb is independently selected from halogen, OH, NH2, —NHC(═O)(C1-C4 alkyl). —NHS(═O)(C1-C4 alkyl), ═O, CN, C1-C4 alkyl and C1-C4alkoxy; p is each independently selected from 0, 1 and 2;
R1X is independently selected from halogen, OH, CN, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, C3-C8 cycloalkyl and cyclic heteroalkyl;
R1B and R1C are independently selected from H, OH, halogen, CN, —(O)—(C1-C10 alkyl comprising 0-2 of substituent Rc), C1-C6 alkyl group, C1-C6 alkoxy, C1-C6 haloalkyl, C1—C haloalkoxy, SCF3, C3-C8 cycloalkyl, —C(═O)(C1-C4 alkyl), —C(═O)NH(C1-C4 alkyl), heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p, —C(═O)H, aryl, 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S; wherein the aryl and heteroaryl may be substituted by 0-2 of R1X; p is each independently 0, 1 and 2;
R2B and R2C are independently selected from H, OH, halogen, CN, —(O)7—(C1-C10 alkyl comprising 0-2 of substituent Rc), C1-C6 alkyl group, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, SCF3, C3-C8 cycloalkyl, —C(═O)(C1-C4 alkyl), —C(═O)NH(C1-C4 alkyl), heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p, —C(═O)H, aryl, 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S; wherein the aryl and heteroaryl may be substituted by 0-2 of R1X; p is each independently 0, 1 and 2;
R3B and R3C are independently selected from H, OH, halogen, CN, —(O)z—(C1-C10 alkyl comprising 0-2 of substituent Rc), C1-C6 alkyl group, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, SCF3, C3-C8 cycloalkyl, —C(═O)(C1-C4 alkyl), —C(═O)NH(C1-C4 alkyl), heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p, —C(═O)H, aryl, 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S; wherein the aryl and heteroaryl may be substituted by 0-2 of R1X; p is each independently 0, 1 and 2;
Alternatively, R1B and R1C, R2B and R2C, R3B and R3C may form a carbonyl group (═O) or a thiocarbonyl group (═S) with a carbon atom to which they are attached;
R1D is independently selected from H, —OH, halogen, CN, —C(═O)H, —(O)7—(C1-C6 alkyl comprising 0-2 of substituent Rc), C1-C6 haloalkyl, C1-C6 haloalkoxy, SCF3, Rf, —ORf, —C(═O)R, NRdRe, —C(═O) NRdRe, —NHC(═O)Rc, —S(═O)2Rc, —S(═O)2, NRdRe, —NHS(═O)2Rd, —(OCH2CH2)mORd, —(OCH2CH2)mNRdRe;
Rf is independently selected from C3-C8 cycloalkyl, heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p, aryl, heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, NRa, O and S(O)p; wherein the aryl and heteroaryl are substituted by 0-2 of R1X;
M is independently selected from a 3 to 7 membered saturated or unsaturated cycloalkyl, a heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p, an aryl, 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S;
In the definition of R1B and R1C, R2B and R2C, R3B and R3C, R1D and Rf, the definitions of Ra, Rc, Rd, Re, p, z, m and R1X are the same as those of R1A in the part 4a);
n is each independently selected from 0, 1 and 2;
m is each independently selected from 0-4;
p is each independently selected from 0-2;
q is each independently selected from 0-3;
z is each independently selected from 0 and 1;
4a′) Preferably, R1 is independently selected from the following structures:
wherein
is a single bond or double bond;
R1A is independently selected from H, hydroxy, halogen, CN, —(O)z(C1-C10 alkyl comprising 0-2 of substituent Re), C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, SCF3, C3-C8 cycloalkyl, —C(═O)(C1-C4 alkyl), —C(═O)NH(C1-C4 alkyl), —C(═O)H;
Rc is independently selected from OH, halogen, CN, C1-C6 alkyl, carboxyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C8 cycloalkyl; R1B and R1C, R2B and R2C, and R3B and R3C are independently selected from H, OH, halogen, CN, —(O)z—(C1-C10 alkyl comprising 0-2 of substituent Rc), C1-C6 alkyl group, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, SCF3, C3-C8 cycloalkyl, —C(═O)(C1-C4 alkyl), —C(═O)NH(C1-C4 alkyl);
Alternatively, R1B and R1C, R2B and R2C, R3B and R1C may form a carbonyl group (═O) or a thiocarbonyl group (═S) with a carbon atom to which they are attached;
R1D is independently selected from H, —OH, halogen, CN. —C(═O)H, —(O)z—(C1-C6 alkyl comprising 0-2 of substituent Rc), C1-C6 haloalkyl, C1-C6 haloalkoxy, SCF3, C3-C8 cycloalkyl;
Rc is independently selected from OH, halogen, CN, C1-C6 alkyl, carboxyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C8 cycloalkyl;
M is independently selected from a 3 to 7 membered saturated or unsaturated cycloalkyl, a heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p, an aryl, 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S;
n is each independently selected from 0, 1 and 2;
m is each independently selected from 0-4;
p is each independently selected from 0-2;
q is each independently selected from 0-3;
z is each independently selected from 0 and 1;
4a″) more preferably, in R1,
R1A is independently selected from H, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C3-C8 cycloalkyl;
R1B and R1C, R2B and R2C, and R1B and R3C are independently selected from H, OH, halogen, C1-C6 alkyl group, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C3-C8 cycloalkyl;
Alternatively, R1B and R1C, R1B and R2C, R3B and R3C may form a carbonyl group (═O) or a thiocarbonyl group (═S) with a carbon atom to which they are attached;
R1D is independently selected from H, —OH, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, C3-C8 cycloalkyl;
M is independently selected from a 5 to 6 membered saturated or unsaturated cycloalkyl, a heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p, an aryl, 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S;
n is each independently selected from 0, 1 and 2;
m is each independently selected from 0-4;
p is each independently selected from 0-2;
q is each independently selected from 0-3;
z is each independently selected from 0 and 1;
4b) Y is independently selected from heteroatoms of O, NRg, S(O)p, etc., and CH2, C═O, —CRi(CH2)mNRgRh and —CRi(CH2)mORg;
Rg and Rh are independently selected from H, O, C1-C10 alkyl comprising 0-3 of substituent Rs, C1-C6 haloalkyl, C3-C6 cycloalkyl,
—C(═S)NHC(═O)—Rj, heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p, aryl, 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S; wherein the aryl and heteroaryl may be substituted by 0-2 of R1X;
Rs is independently selected from OH, CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C8 cycloalkyl, —(OCH2CH2)mORd, NHC(═O)NRdRe, NHC(═S)NRdRe, —NHC(═NH)NRdRe, (OCH2CH2)mNRdRe, —C(═O)Rd, —C(═S)Rd, —S(═O) Rd, —C(═O)NRdRe, —S(═O)2Rd, —NHC(═O)Rd, —NHC(═S)Rd. —NHS(═O)2Rd, —S(═O)2NRdRe, —NHS(═O)2NRdRe, —C(═S)NRdRe, NHC(═O)ORd, NHC(═S)ORd, —NHS(═O)2ORd, NHC(═O)SRd, NHC(═S)SRd, —NHC(═NH)ORd, —C(═O)ORd, —C(═O)SRd, —S(═O)2ORd, heteroalkyl and heterocycloalkyl comprising carbon atoms and 1 to 2 heteroatoms selected from N, NRa, O and S(O)p, aryl, heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, NRa, O and S(O)p; wherein the aryl and heteroaryl may be substituted by 0-2 of R1Y;
Rd and Re are independently selected from H, C1-C6 alkyl comprising 0-2 of Rb, C1-C6 haloalkyl, C3-C6 cycloalkyl, —C(═O)(C1-C4 alkyl), —CO2(C1-C4 alkyl), —C(═O)NH(C1-C4 alkyl), C1-C6 branched or cyclic heteroalkyl comprising 0-2 of heteroatoms selected from O, N, and S(O)p, —C(═O)H, aryl, and heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, NRa, O and S(O)p, wherein the aryl and heteroaryl may be substituted by 0-2 of R1X;
Rd and Re may be connected by the following manner
to form a ring, wherein Z1 can be selected from C1-C6 alkyl comprising from 0-2 of substituent Rb; C1-C6 alkyl comprising 0-2 heteroatoms of O, N, S(O)p, O; —N(C1-C6 alkyl); —NH; —N(C═O) C1-C6 alkyl; —NS(═O)2(C1-C6 alkyl); S(O)p;
Each Ra is independently selected from H; O; C1-C10 alkyl substituted by 0-2 of Rb; C1-C6 haloalkyl; —O—(C1-C6 alkyl); C1-C6 haloalkoxy; C3-C6 cycloalkyl; —C(═O)(C1-C4 alkyl); —CO2(C1-C4 alkyl); heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p; —C(═O)H; aryl; 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S;
Rb is independently selected from halogen, OH, NH2, NHC(═O)(C1-C4 alkyl), NHS(═O)2(C1-C4 alkyl), ═O, CN, C1-C4 alkyl and C1-C4 alkoxy; p is independently selected from 0.1 and 2;
R1X is independently selected from halogen, OH, CN, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, C3-C8 cycloalkyl and cyclic heteroalkyl;
R1Y is independently selected from C1-C10 alkyl; halogen; CN; —(O)2—(C1-C10 alkyl comprising 0-2 of the substituent Re); C1-C6 haloalkyl; C1-C6 haloalkoxy; SCF3; C3-C8 cycloalkyl; —C(═O)(C)—C4 alkyl); —CO2(C1-C4 alkyl); NRdRe; —C(═O)NRdRe; —S(═O)2Rd; —NHC(═O)Rd; —NHC(═S)Rd; —NHS(═O)2Rd; —NHC(═O)NRdRe; —NHC(═S)NRdRe; —NHS(═O)2NRdRe; —C(═S)NRdRe; —S(═O)2NHRd; heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p; —C(═O)H; p is each independently 0, 1 and 2;
wherein, Rc is the same definition of R as defined in the above part 4a);
wherein, Rd and Re may be connected by the manner of
wherein Z1 can be selected from C1-C6 alkyl comprising from 0-2 of substituent Rb; C1-C6 heteroalkyl comprising 0-2 heteroatoms of O, N, S(O)P; O; —N(C1-C6 alkyl); —NH; —N(C═O) C1-C6 alkyl; —NS(═O)2(C1-C6 alkyl); S(O)p;
Rb is independently selected from halogen, OH, NH2. NHC(═O)(C1-C4 alkyl), NHS(═O)2(C1-C4 alkyl), ═O, CN, C1-C4 alkyl and C1-C4 alkoxy; p is independently selected from 0, 1 and 2;
Rj and Rk are independently selected from H, CN, C1-C10 alkyl comprising 0-3 of substituent R5, C1-C6 haloalkyl, C3-C10 cycloalkyl, heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p, alkenyl or alkynyl group substituted by Ry
6 to 10 membered aryl, 5 to 10 membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O, and S; wherein the aryl and heteroaryl groups may be substituted by 0-2 of R1Y;
wherein, R1Y is the same definition of R1Y as defined in the above R5 in the part 4b);
Ry is independently selected from H; C1-C10 alkyl comprising 0-3 of substituent Re; C1-C6 haloalkyl; C3-C10 cycloalkyl; heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p; NRdRe; ORd aryl; 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S; wherein the aryl and heteroaryl may be substituted by 0-2 of R1X; R1X is independently selected from halogen, OH, CN, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, C3-C8 cycloalkyl and cyclic heteroalkyl;
wherein, Re is the same definition of R as defined in the above part 4a); Rd and Re are the same definition of Rd and Re as defined in the above R5 in the part 4b);
In particular, Rg and Rh, as well as Rj and Rk may be connected by the following manner
wherein Z1 may be selected from C1-C6 alkyl comprising 0-2 of substituent Rc; C1-C6 alkyl comprising 0-2 heteroatoms of O, N, S(O)p; O; —N(C1-C6 alkyl); —NH; —N(C═O) C1-C6 alkyl; —NS(═O)2(C1-C6 alkyl); S(O)p; p is each independently selected from 0, 1 and 2;
wherein, Rc is the same definition of Rc as defined in the above part 4a);
Ri is independently selected from H, CN, C1-C4 alkyl;
m is each independently selected from 0-4:
4b′) Preferably, Y is independently selected from O, NR5, S(O)p, —CRi(CH2)mNRgRh and —CRi(CH2)mORg;
Rg and Rh are independently selected from H; C1-C6 haloalkyl;
—C(═S)NHC(═O)—Rj; heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p; aryl; 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S; wherein the aryl and heteroaryl may be substituted by 0-2 of R1X;
Rj and Rk are independently selected from H; CN; C1-C10 alkyl comprising 0-3 of substituent R5; C1-C6 haloalkyl; C3-C10 cycloalkyl; heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S(O)p; C2-C10 alkenyl or alkynyl; 6 to 10 membered aryl; 5 to 10 membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O, and S; wherein the aryl and heteroaryl may be substituted by 0-2 of R1Y:
wherein, the definition of Rs is the same definition of Rs in the above part 4b);
p is each independently selected from 0, 1 and 2;
R1X is independently selected from halogen, OH, CN, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, C3-C8 cycloalkyl and cyclic heteroalkyl;
R1Y is independently selected from C1-C10 alkyl, halogen, CN, C1-C6 haloalkyl, C1-C6 haloalkoxy, C3-C8 cycloalkyl;
p is each independently 0, 1 and 2;
In particular, Rg and Rh, as well as Rj and Rk may be connected by the following manner
wherein Z1 may be selected from C1-C6 alkyl comprising 0-2 of substituent Rc; C1-C6 alkyl comprising 0-2 heteroatoms of O, N, S(O)p; O; —N(C1-C6 alkyl); —NH; —N(C═O) C1-C6 alkyl; —NS(═O)2(C1-C6 alkyl); S(O)p;
wherein, Rc is the same definition of R as defined in the above part 4a);
p is each independently selected from 0, 1 and 2:
R1 is independently selected from H, CN and C1-C4 alkyl;
m is each independently selected from 0-4;
4b″) More preferably, the Y is independently selected from O, NRg, S, —CRiNRgRh and —CRiORg;
Rg and Rh are independently selected from H; C1-C6 haloalkyl;
—C(═S)NHC(═O)—Rj; heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S; aryl; 5- to 6-membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O and S; wherein the aryl and heteroaryl may be substituted by 0-2 of R1X;
Rj and Rk are independently selected from H; CN; C1-C10 alkyl comprising 0-3 of substituent Rs; C1-C6 haloalkyl; C3-C10 cycloalkyl; heteroalkyl and heterocycloalkyl comprising carbon atoms and 1-4 heteroatoms selected from O, N, S; C2-C10 alkenyl or alkynyl; 6 to 10 membered aryl; 5 to 10 membered heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O, and S; wherein the aryl and heteroaryl may be substituted by 0-2 of R1Y;
Rs is independently selected from OH; CN; halogen; C1-C6 alkyl; C1-C6 haloalkyl; C1-C6 alkoxy; C1-C6 haloalkoxy; C3-C8 cycloalkyl, —(OCH2CH2)mORd, (OCH2CH2)mNRdRe, heteroalkyl and heterocycloalkyl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O, S; aryl; and heteroaryl comprising carbon atoms and 1 to 2 heteroatoms selected from N, O, and S; wherein the aryl and heteroaryl may be substituted by 0-2 of R1Y;
wherein, Rd, Re are the same definition of Rd, Re as defined in the above part 4b);
R1Y is independently selected from C1-C10 alkyl, halogen, CN, C1-C6 haloalkyl, C1-C6 haloalkoxy, C3-C8 cycloalkyl:
In particular, Rg and Rh, as well as Rj and Rk may be connected by the following manner
wherein Z1 may be selected from C1-C6 alkyl comprising 0-2 of substituent Re; C1-C6 alkyl comprising 0-2 heteroatoms of O, N, S(O)p; O; —N(C1-C6 alkyl); —NH; —N(C═O) C1-C6 alkyl; —NS(═O)2(C1-C6 alkyl); S(O)p;
wherein, Rc is the same definition of Rc as defined in the above part 4a):
p is each independently selected from 0, 1 and 2:
R1 is independently selected from H, CN and C1-C4 alkyl;
m is each independently selected from 0-4;
Preferably, the compound of formula I has formula Ia-1 or Ia-2;
Wherein, X1 is the same definition as defined in the part 1) of Formula I:
is a single bond or double bond;
X2, R3-R5, R6 and n are the same definition as defined in the part 3) of Formula I;
R1A, R1B and R1C, R2B and R2C, R3B and R3C, R1D, n, n, q, Y, M are the same definition as defined in the part 4) of Formula I;
Preferably, the compound of formula I has formula Ia-3 or Ia-4:
Wherein, X1 is the same definition as defined in the part 1) of Formula I;
is a single bond or double bond;
X2, R7 and n are the same definition as defined in the part 3) of Formula I;
R1A, R1B and R1C, R1B and R2C, R3B and R3C, R1D, n, m, q, Y, M are the same definition as defined in the part 4) of Formula I;
Preferably, the compound of formula I has formula Ia-5 or Ia-6:
Wherein, X1 is the same definition as defined in the part 1) of Formula I;
is a single bond or double bond;
Y is the same definition as defined in the part 4) of Formula I:
Preferably, the compound of formula I has formula Ia-7 or Ia-8:
Wherein, X1 is the same definition as defined in the part 1) of Formula I;
is a single bond or double bond;
The definition of Rg is the same definition as defined in the part 4b) of part 4) of Formula I;
Preferably, the compound of formula I has formula Ia-9:
wherein
is a single bond or double bond;
The definition of Rg is the same definition as defined in the part 4b) of part 4) of Formula I:
Preferably, the compound of formula I has one of the following formulae:
wherein
M is the same definition as defined in the part 4) of Formula I;
The definition of Rg is the same definition as defined in the part 4b) of part 4) of Formula I;
Preferably, the compound of formula I is selected from the following compounds:
Preferably, the compound also includes the stereoisomers, tautomers, atropisomers, isotopically labeled compounds (including deuterium substitution), medically acceptable salts, polymorphs, solvates thereof, which can be used to treat diseases or conditions mediated by EED and/or PRC2.
According to another aspect of the invention, methods and intermediates for preparing the compounds of the invention are provided.
Wherein, the method comprises the following steps:
(1a) treating 5-bromo-4-chloro-2-(methylthio) pyrimidine 1 with hydrated hydrazine to produce 5-bromo-4-hydrazinyl-2-(methylthio) pyrimidine 2,
(1b) converting 5-bromo-4-hydrazinyl-2-(methylthio) pyrimidine 2 with trimethyl orthoformate to triazole product 3,
(1c) conducting a substitution reaction of triazole product 3 with an amine NH2CH2A to produce compound 4,
(1d) conducting a suzuki coupling reaction of compound 4 with various of boric acid having R1 group or its equivalent under the action of palladium catalyst to obtain product 5.
wherein, the definitions of A, R1 are the same as defined above.
(2a) reacting the cyanoethyl amide 6 with ethyl propiolate to produce intermediate 7,
(2b) treating intermediate 7 with bromine to conduct a bromation reaction to obtain bromide 8,
(2c) reacting bromide 8 with phosphorus oxychloride to obtain intermediate 9,
(2d) treating intermediate 9 with hydrated hydrazine to produce intermediate 10,
(2e) converting intermediate 10 with trimethyl orthoformate to triazole intermediate 11,
(2f) conducting a substitution reaction of triazole intermediate 11 with various amines to produce compound 12,
(2g) conducting a suzuki coupling reaction of compound 12 with various of boric acid having R1 group or its equivalent under the action of palladium catalyst to obtain product 13.
wherein, the definitions of A, R1 are the same as defined above.
The amines described in Scheme 1 and Scheme 2 can be prepared according to the patent US2016/0176682A1 (such as the preparation of A1 in the following reaction formula), or can be purchased from a reagent company (the following reaction formula, furfurylamine A2, purchased from Bellingway Technology Co., Ltd.), the boric acid or its equivalent B may be purchased from a reagent company or may be prepared according to conventional literature.
(3a) removing the Boc protecting group in 14 using dichloromethane as a solvent and under the action of trifluoroacetic acid to obtain an amine compound 15,
(3b) reacting the amine compound 15 with a reagent or compound having Rg under basic conditions to give a compound 16, the agent or compound is, for example, but not limited to acid anhydrides, sulfonic anhydride, isocyanate, thioisocyanate, acyl chloride, sulfonyl chloride, carbonate, chloroformate, urethane, etc., the base is, for example, but not limited to, triethylamine, diisopropylethylamine, DMAP, potassium carbonate, sodium hydroxide, potassium hydroxide, potassium tert-butoxide, NaH, the organic solvents is, for example, but not limited to, methylene chloride, tetrahydrofuran, acetonitrile, 1,4-dioxanene,
wherein, the definitions of A, X1, R8, R1A, R1B, R1C, R2B, R2C, R3B, R3C, q, m are the same as defined above.
(4a) conducting a condensation reaction of product 15 obtained by removing the protective group in step (3a) of Scheme 3 with a carboxylic acid having an Rj group under the action of a condensing agent to obtain an amide compound 17, the condensing agent is, for example, but not limited to carbonyldiimidazole, dicyclohexylcarbodiimide, diisopropylcarbodiimide, 1-(-3-dimethylaminopropyl)-3-ethylcarbodiimide, 1-hydroxybenzotriazole, 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate, benzotriazole-N,N,N′,N′-tetramethylurea hexafluorophosphate, 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate, o-benzotriazole-N,N,N′,N′-tetramethylurea tetrafluoroborate, 6-chlorobenzotriazole-1,1,3,3-tetramethylurea tetrafluoroborate, 2-succinimidyl-1,1,3,3-tetramethylurea tetrafluoroborate and 2-(5-norbornene-2,3-dicarboximido)-1,1,3,3-tetramethylurea quaternary ammonium tetrafluoroborate, the condensation reaction can be performed in an organic solvent in the presence of a base, the base is, for example, but not limited to, triethylamine, diisopropylethylamine, 1,5-diazabicyclo[5.4.0]undec-5-ene, the organic solvent is, for example, but not limited to dichloromethane, chloroform, N,N-dimethylformamide, tetrahydrofuran,
wherein, the definitions of A, X1, Rj, R1A, R1B, R1C, R2B, R2C, R3B, R3C, q, m are the same as defined above.
(5a) dissolving 18 in a solvent, the solvent is, for example, but not limited to, methanol, ethanol, ethyl acetate, and tetrahydrofuran, adding a metal catalyst, the metal catalyst is, for example, but not limited to 10% palladium carbon. Pd(OH)2, Raney nickel, RhCl(PPh3)3, introducing hydrogen gas, and reacting at room temperature to obtain compound 19 with double bond reduction.
wherein, the definitions of A, X1, Y, R1A, R1B, R1C, R2B, R2C, R3B, R3C, q, m are the same as defined above.
(6a) obtaining the compound 21 by reduction reaction of 20, and then conducting an oxidation reaction with mCPBA (m-chloroperoxybenzoic acid) or hydrogen peroxide to obtain compound 22.
wherein, the definitions of A, X1, R1A, R1B, R1C, R2B, R2C, R3B, R3C, q, m are the same as defined above.
(7a) conducting an oxidation reaction of 20 with mCPBA or hydrogen peroxide to obtain compound 23.
wherein, the definitions of A, X1, R1A, R1B, R1C, R2B, R2C, R3B, R3C, q, m are the same as defined above.
Reducing (for example, but not limited to, under the conditions of hydrogenation reduction) the double bond of 15 to obtain 24, and then reacting with a reagent or compound having an R9 group in the presence of a base to obtain 25, said reagent or compound is, for example, but not limited to acid anhydrides, sulfonic anhydride, isocyanate, thioisocyanate, acyl chloride, sulfonyl chloride, carbonate, chloroformate, urethane, the base is, for example, but not limited to, triethylamine, diisopropylethylamine. DMAP, potassium carbonate, sodium hydroxide, potassium hydroxide, potassium tert-butoxide, NaH, the reaction can be performed in an organic solvent is, for example, but not limited to, methylene chloride, tetrahydrofuran, acetonitrile, 1,4-dioxane; or conducting a condensation reaction of 24 with various carboxylic acids in the presence of a condensing agent to obtain an amide compound 25, the condensing agent is, for example, but not limited to carbonyldiimidazole, dicyclohexylcarbodiimide, diisopropylcarbodiimide, 1-(-3-dimethylaminopropyl)-3-ethylcarbodiimide, 1-hydroxybenzotriazole, 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate, benzotriazole-N,N,N′,N′-tetramethylurea hexafluorophosphate, 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate, o-benzotriazole-N,N,N′,N′-tetramethylurea tetrafluoroborate, 6-chlorobenzotriazole-1,1,3,3-tetramethylurea tetrafluoroborate, 2-succinimidyl-1,1,3,3-tetramethylurea tetrafluoroborate and 2-(5-norbornene-2,3-dicarboximido)-1,1,3,3-tetramethylurea quaternary ammonium tetrafluoroborate, the condensation reaction can be performed in an organic solvent in the presence of a base, the base is triethylamine, diisopropylethylamine, 1,5-diazabicyclo[5.4.0]undec-5-ene, the organic solvent is dichloromethane, chloroform, N,N-dimethylformamide, tetrahydrofuran,
wherein, the definitions of A, X1, R1A, R1B, R1C, R2B, R2C, R3B, R3C, q, m are the same as defined above.
Preferably, the method comprises the following steps:
(1a) treating 5-bromo-4-chloro-2-(methylthio) pyrimidine 1 with hydrated hydrazine to produce 5-bromo-4-hydrazinyl-2-(methylthio) pyrimidine 2,
(1b) converting 5-bromo-4-hydrazinyl-2-(methylthio) pyrimidine 2 with trimethyl orthoformate to triazole product 3,
(1c) conducting a substitution reaction of triazole product 3 with an amine NH2CH2A to produce compound 4,
(1d) conducting a suzuki coupling reaction of compound 4 with various of boric acid having R1 group or its equivalent under the action of palladium catalyst to obtain product 5.
wherein, the definitions of A, R1 are the same as defined above;
(2a) reacting the cyanoethyl amide 6 with ethyl propiolate to produce intermediate 7,
(2b) treating intermediate 7 with bromine to conduct a bromation reaction to obtain bromide 8,
(2c) reacting bromide 8 with phosphorus oxychloride to obtain intermediate 9,
(2d) treating intermediate 9 with hydrated hydrazine to produce intermediate 10,
(2e) converting intermediate 10 with trimethyl orthoformate to triazole product 11,
(2f) conducting a substitution reaction of triazole product 11 with various amines to produce compound 12,
(2g) conducting a suzuki coupling reaction of compound 12 with various of boric acid having R1 group or its equivalent under the action of palladium catalyst to obtain product 13,
wherein, the definitions of A, R1 are the same as defined above;
(3a) removing the Boc protecting group in 14′ using dichloromethane as a solvent and under the action of trifluoroacetic acid to obtain an amine compound 15′.
(3b) further reacting the amine compound 15′ with an acid anhydride, a sulfonic anhydride, an isocyanate, and a thioisocyanate with an R9 group under basic conditions to obtain a compound 16′, the base is triethylamine, diisopropylethylamine, the reaction may be performed in an organic solvent, the organic solvent is dichloromethane, tetrahydrofuran, acetonitrile, 1,4-dioxane,
wherein, the definitions of A, X1, Rg are the same as defined above:
(4a) conducting a condensation reaction of product 15′ obtained by removing the protective group in step (3b) of Scheme 3 with a carboxylic acid having an Rj group under the action of a condensing agent to obtain an amide compound 17′, the condensing agent is selected from carbonyldiimidazole, dicyclohexylcarbodiimide, diisopropylcarbodiimide, 1-(-3-dimethylaminopropyl)-3-ethylcarbodiimide, 1-hydroxybenzotriazole, 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate, benzotriazole-N,N,N′,N′-tetramethylurea hexafluorophosphate, 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate, o-benzotriazole-N,N,N′,N′-tetramethylurea tetrafluoroborate, 6-chlorobenzotriazole-1,1,3,3-tetramethylurea tetrafluoroborate, 2-succinimidyl-1,1,3,3-tetramethylurea tetrafluoroborate and 2-(5-norbornene-2,3-dicarboximido)-1,1,3,3-tetramethylurea quaternary ammonium tetrafluoroborate, the condensation reaction can be performed in an organic solvent in the presence of a base, the base is triethylamine, diisopropylethylamine, 1,5-diazabicyclo[5.4.0]undec-5-ene, the organic solvent is dichloromethane, chloroform, N,N-dimethylformamide, tetrahydrofuran,
wherein, the definitions of A, X1, Rj are the same as defined above;
(5a) using methanol as a solvent, and catalyzed by 10% palladium on carbon, introducing hydrogen gas, reacting 18′ at room temperature for 6 hours to obtain a compound 19′ with the double bond reduced.
wherein, the definitions of X1, Y are the same as defined above. The above-mentioned compounds 14, 14′, 18, 18′ and 20 can be obtained by a Suzuki coupling reaction according to step (1d) or step (2g) in Scheme 1 or Scheme 2.
When an optically active form of a compound of the invention is required, it can be obtained by using optically active starting materials, or can be obtained by resolving the mixture of stereoisomers of the compound or intermediate by using standard procedures known to those skilled in the art, such as separation by a chiral chromatography column.
Similarly, when pure geometric isomers of the compounds of the present invention are required, they can be obtained by using pure geometric isomers as starting materials, or can be obtained by resolving the mixture of geometric isomers of compounds or intermediates by using standard procedures, such as chromatographic separation.
According to another aspect of the present invention, a pharmaceutical composition is provided, which comprises one or more of triazolopyrimidine, triazolopyridine compounds, pharmaceutically acceptable salts, enantiomers, diastereomers or racemates.
Preferably, the pharmaceutical composition further comprises at least one other therapeutic agent.
Preferably, at least one other therapeutic agent included in the pharmaceutical composition is selected from other anticancer agents, immunomodulators, antiallergic agents, antiemetics, pain relief agents, cytoprotective agents, and combinations thereof.
Preferably, the pharmaceutical composition comprises at least one compound of the present invention and at least one pharmaceutically acceptable carrier, diluent or excipient,
According to another aspect of the present invention, the use of the above compound or the pharmaceutical composition in the preparation of a medicament for treating a disease or condition mediated by EED and/or PRC2 is provided.
Preferably, the disease or condition includes diffuse large B-cell lymphoma, follicular lymphoma, other lymphoma, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, hepatocellular carcinoma, prostate Cancer, breast cancer, bile duct and gallbladder cancer, bladder cancer; brain tumors, including neuroblastoma, schwannoma, glioma, glioblastoma and astrocytoma; cervical cancer, colon cancer, melanin tumor, endometrial cancer, esophageal cancer, head and neck cancer, lung cancer, nasopharyngeal cancer, ovarian cancer, pancreatic cancer, renal cell cancer, rectal cancer, thyroid cancer, parathyroid tumor, uterine tumor and soft tissue sarcoma.
According to another aspect of the invention, a method of treating a disease or condition mediated by EED and/or PRC2 is provided, the method comprising
Providing a subject in need with a therapeutically effective amount of a compound of formula I or a pharmaceutical composition thereof.
Preferably, the disease or condition is selected from diffuse large B-cell lymphoma, follicular lymphoma, other lymphoma, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, hepatocellular carcinoma, prostate Cancer, breast cancer, bile duct and gallbladder cancer, bladder cancer; brain tumors, including neuroblastoma, schwannoma, glioma, glioblastoma and astrocytoma; cervical cancer, colon cancer, melanin tumor, endometrial cancer, esophageal cancer, head and neck cancer, lung cancer, nasopharyngeal cancer, ovarian cancer, pancreatic cancer, renal cell cancer, rectal cancer, thyroid cancer, parathyroid tumor, uterine tumor and soft tissue sarcoma.
All features disclosed in this specification, or the disclosed methods or steps can be combined in any way unless that the features and/or steps are mutually exclusive. The following examples are only used to explain partial scope of the present invention and are not intended to limit the protection scope of the present invention.
Bromide 4-1 (72 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-1 (84 mg, 0.4 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y1 as a white solid (41 mg, 57%).
1H NMR (400 MHz, MeOD-d4) δ 9.33 (s, 1H), 7.83 (s, 1H), 6.96 (s, 1H), 6.90-6.84 (m, 1H), 6.69-6.63 (m, 1H), 4.81 (s, 2H), 4.59 (t, J=8.7 Hz, 2H), 4.39 (m, 2H), 3.99 (t, J=5.5 Hz, 2H), 3.38 (m, 2H), 2.62 (m, 2H). LC-MS: [M+H]+=368.1.
Bromide 4-1 (72 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-2 (90 mg, 0.4 mmol) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y2 as a white solid (34 mg, 45%).
1H NMR (400 MHz, DMSO-d6) δ 9.49 (s, 1H), 8.70 (t, J=4.7 Hz, 1H), 7.67 (s, 1H), 7.26 (s, 1H), 6.94 (t, J=9.5 Hz, 1H), 6.70 (dd, J=8.6, 3.9 Hz, 1H), 4.68 (d, J=4.6 Hz, 2H), 4.53 (t, J=8.7 Hz, 2H), 3.36 (m, 2H), 3.29 (t, J=8.6 Hz, 2H), 2.86 (t, J=5.6 Hz, 2H), 2.74 (m, 2H). LC-MS: [M+H]+=384.1.
Bromide 4-1 (72 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-3 (123 mg, 0.4 mmol) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y3 as a white solid (67 mg, 72%).
1H NMR (400 MHz, MeOD-d4) δ 9.36 (s, 1H), 7.88 (s, 1H), 6.91-6.82 (m, 1H), 6.73 (s, 1H), 6.66 (dd, J=8.7, 3.9 Hz, 1H), 4.81 (s, 2H), 4.59 (t, J=8.7 Hz, 2H), 4.16 (s, 2H), 3.76-3.66 (m, 2H), 3.38 (t, J=8.7 Hz, 2H), 2.62 (s, 2H), 1.52 (s, 9H). LC-MS: [M+H]+=467.2.
Bromide 4-2 (58 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-1 (84 mg, 0.4 mmol) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y4 as a white solid (37 mg, 64%).
1H NMR (400 MHz, MeOD-d4) δ 9.24 (s, 1H), 7.70 (s, 1H), 7.46 (d, J=0.9 Hz, 1H), 7.06 (s, 1H), 6.44-6.31 (m, 2H), 4.77 (s, 2H), 4.37 (m, 2H), 3.96 (t, J=5.5 Hz, 2H), 2.60 (m, 2H). LC-MS: [M+H]+=298.1.
Bromide 4-2 (58 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-2 (90 mg, 0.4 mmol) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y5 as a white solid (37 mg, 60%).
1H NMR (400 MHz, MeOD-d4) δ 9.25 (s, 1H), 7.68 (s, 1H), 7.48 (d, J=0.9 Hz, 1H), 6.85-6.81 (m, 1H), 6.42-6.36 (m, 2H), 4.79 (s, 2H), 3.43-3.37 (m, 2H), 2.93 (t, J=5.7 Hz, 2H), 2.81 (dd, J=5.7, 2.0 Hz, 2H). LC-MS: [M+H]+=314.1.
Bromide 4-2 (58 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-3 (123 mg, 0.4 mmol) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y6 as a white solid (55 mg, 70%).
1H NMR (400 MHz, MeOD-d4) δ 9.24 (s, 1H), 7.68 (s, 1H), 7.47 (dd, J=1.8, 0.8 Hz, 1H), 6.94 (s, 1H), 6.40 (ddd, J=5.1, 3.2, 2.2 Hz, 2H), 4.78 (s, 2H), 4.15 (m, 2H), 3.69 (m, 2H), 2.62 (m, 2H), 1.51 (s, 9H). LC-MS: [M+H]+=397.1.
Bromide 4-2 (58 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-4 (83 mg, 0.4 mmol) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y7 as a white solid (38 mg, 64%).
1H NMR (400 MHz, MeOD-d4) δ 9.24 (s, 1H), 7.66 (s, 1H), 7.49 (s, 1H), 6.83 (s, 1H), 6.44-6.36 (m, 2H), 4.79 (s, 2H), 2.52 (s, 2H), 2.31 (s, 2H), 1.87 (dd, J=11.7, 6.1 Hz, 2H), 1.79-1.70 (m, 2H). LC-MS: [M+H]+=296.1.
Bromide 4-2 (58 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-5 (102 mg, 0.4 mmol) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y8 as a white solid (39 mg, 57%).
1H NMR (400 MHz, MeOD-d4) δ 9.24 (s, 1H), 7.65 (s, 1H), 7.48 (d, J=1.0 Hz, 1H), 7.20 (d, J=6.9 Hz, 1H), 7.12 (t, J=6.9 Hz, 1H), 7.03 (t, J=7.0 Hz, 1H), 6.80 (d, J=7.3 Hz, 1H), 6.46-6.38 (m, 2H), 6.34 (t, J=4.6 Hz, 1H), 4.81 (s, 2H), 2.91 (t, J=8.0 Hz, 2H), 2.46 (td, J=8.0, 4.7 Hz, 2H). LC-MS: [M+H]+=344.1.
Bromide 4-2 (58 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-6 (129 mg, 0.4 mmol) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y9 as a white solid (42 mg, 51%).
1H NMR (400 MHz, MeOD-d4) δ 9.23 (s, 1H), 7.62 (s, 1H), 7.47 (dd, J=1.8, 0.9 Hz, 1H), 6.76 (s, 1H), 6.39 (ddd, J=5.1, 3.2, 1.3 Hz, 2H), 4.77 (s, 2H), 3.75 (m, 1H), 2.68-2.51 (m, 3H), 2.25-2.14 (m, 1H), 2.04 (m, 1H), 1.80-1.68 (m, 1H), 1.46 (s, 9H). LC-MS: [M+H]+=411.2.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After being concentrated, the resultant was directly separated by HPLC. Mobile phase: 40% acetonitrile/60% water (containing 0.1% TFA), retention time 4.7 min, yield 70%.
LC-MS: [M+H]+=367.1.
Compound E-Y6 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After being concentrated, the resultant was directly separated by HPLC. Mobile phase: 40% acetonitrile/60% water (containing 0.1% TFA), retention time 5.4 min, yield 82%.
1H NMR (400 MHz, MeOD-d4) δ 9.27 (s, 1H), 7.77 (s, 1H), 7.45 (dd, J=1.8, 0.8 Hz, 1H), 7.02 (s, 1H), 6.38 (dt, J=3.2, 2.2 Hz, 2H), 4.79 (s, 2H), 3.92 (m, 2H), 3.50 (t, J=6.1 Hz, 2H), 2.95-2.85 (m, 2H). LC-MS: [M+H]+=297.1.
Compound E-Y9 (41 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After being concentrated, the resultant was directly separated by HPLC. Mobile phase: 40% acetonitrile/60% water (containing 0.1% TFA), retention time 5.4 min, yield 69%.
1H NMR (400 MHz, MeOD-d4) δ 9.29 (s, 1H), 7.79 (s, 1H), 7.46 (s, 1H), 6.61 (s, 1H), 6.45-6.34 (m, 2H), 4.79 (s, 2H), 3.51 (m, 1H), 2.70 (m, 2H), 2.41-2.32 (m, 1H), 2.20 (m, 1H), 1.91 (m, 2H). LC-MS: [M+H]+=311.1.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. methanesulfonic anhydride (Ms2O, 26 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y13 as a white solid (24 mg, yield in two steps 51%).
1H NMR (400 MHz, DMSO-d6) δ 9.46 (s, 1H), 8.67 (t, J=4.9 Hz, 1H), 7.71 (s, 1H), 7.32 (s, 1H), 7.01-6.88 (m, 1H), 6.70 (dd, J=8.6, 4.0 Hz, 1H), 4.69 (d, J=4.8 Hz, 2H), 4.54 (t, J=8.8 Hz, 2H), 3.94 (m, 2H), 3.40 (m, 2H), 3.29 (m, 2H), 2.95 (s, 3H), 2.70 (m, 2H). LC-MS: [M+H]+=445.1
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. p-toluenesulfonic acid anhydride (Ts2O, 48 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y14 as a white solid (16 mg, yield in two steps 32%).
1H NMR (400 MHz, MeOD-d4) δ 9.45 (s, 1H), 7.89-7.64 (m, 3H), 7.44 (d, J=7.9 Hz, 2H), 6.85 (t, J=9.4 Hz, 1H), 6.76 (s, 1H), 6.64 (dd, J=8.5, 3.6 Hz, 1H), 4.77 (s, 2H), 4.57 (t, J=8.6 Hz, 2H), 3.79 (s, 2H), 3.44-3.33 (m, 4H), 2.67 (s, 2H), 2.45 (s, 3H). LC-MS: [M+H]+=521.2.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Acetic anhydride (Ac2O, 15 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y15 as a white solid (14 mg, yield in two steps 35%).
1H NMR (400 MHz, DMSO-d6) δ 9.46 (d, J=1.1 Hz, 1H), 8.65 (s, 1H), 7.70 (d, J=8.3 Hz, 1H), 7.27 (d, J=21.1 Hz, 1H), 6.97-6.89 (m, 1H), 6.71 (dd, J=8.7, 3.8 Hz, 1H), 4.69 (d, J=4.2 Hz, 2H), 4.54 (t, J=8.7 Hz, 2H), 4.19 (d, J=25.2 Hz, 2H), 3.67 (dt, J=11.2, 5.7 Hz, 2H), 3.28 (t, J=8.7 Hz, 2H), 2.64 (s, 1H), 2.52 (s, 1H), 2.06 (d, J=15.7 Hz, 3H). LC-MS: [M+H]+=409.1.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Benzoic anhydride ((PhCO)2O, 34 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y16 as a white solid (14 mg, yield in two steps 30%).
1H NMR (400 MHz, DMSO-d6) δ 9.46 (s, 1H), 8.66 (s, 1H), 7.70 (s, 1H), 7.47 (d, J=3.9 Hz, 5H), 7.01-6.90 (m, 1H), 6.71 (dd, J=8.7, 3.9 Hz, 1H), 4.69 (d, J=4.9 Hz, 2H), 4.54 (t, J=8.8 Hz, 2H), 4.35 (s, 1H), 4.15 (s, 1H), 3.87 (s, 1H), 3.56 (s, 1H), 3.28 (t, J=8.6 Hz, 2H), 2.64 (s, 2H). LC-MS: [M+H]+=471.1.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Ethyl isocyanate (11 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y17 as a white solid (17 mg, yield in two steps 40%).
1H NMR (400 MHz, DMSO-d6) δ 9.45 (s, 1H), 8.64 (s, 1H), 7.69 (s, 1H), 7.27 (s, 1H), 6.95 (t, J=9.2 Hz, 1H), 6.78-6.64 (m, 1H), 6.51 (s, 1H), 4.69 (s, 2H), 4.54 (t, J=8.8 Hz, 2H), 4.03 (s, 2H), 3.54 (s, 2H), 3.27 (m, 2H), 3.15-2.98 (m, 2H), 1.03 (t, J=7.1 Hz, 3H). LC-MS: [M+H]t=438.2.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring, t-butyl isocyanate (11 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y18 as a white solid (15 mg, yield in two steps 33%).
1H NMR (400 MHz, MeOD-d4) δ 9.38 (s, 1H), 7.93 (s, 1H), 6.93-6.83 (m, 1H), 6.69 (s, 1H), 6.66 (dd, J=8.7, 3.9 Hz, 1H), 4.82 (s, 2H), 4.59 (t, J=8.7 Hz, 2H), 4.10 (d, J=2.9 Hz, 2H), 3.66 (t, J=5.6 Hz, 2H), 3.39 (t, J=7.7 Hz, 2H), 2.62 (s, 2H), 1.38 (s, 9H). LC-MS: [M+H]+=466.2.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Ethyl isothiocyanate (13 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y19 as a white solid (14 mg, yield in two steps 31%).
1H NMR (400 MHz, DMSO-d6) δ 9.46 (s, 1H), 8.66 (d, J=5.0 Hz, 1H), 7.71 (d, J=9.9 Hz, 2H), 7.27 (s, 1H), 7.02-6.90 (m, 1H), 6.71 (dd, J=8.5, 3.8 Hz, 1H), 4.70 (d, J=4.7 Hz, 2H), 4.54 (t, J=8.7 Hz, 2H), 4.41 (s, 2H), 4.07 (t, J=5.4 Hz, 2H), 3.55 (dd, J=12.3, 6.8 Hz, 2H), 3.27 (d, J=8.7 Hz, 2H), 2.61 (s, 2H), 1.12 (t, J=7.1 Hz, 3H). LC-MS: [M+H]+=454.1.
Compound E-Y6 (39 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Acetic anhydride (Ac2O, 15 mg, 0.15 mmol) was added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y20 as a white solid (14 mg, yield in two steps 42%).
1H NMR (400 MHz, MeOD-d4) δ 9.33 (s, 1H), 7.91 (s, 1H), 7.49 (s, 1H), 6.73 (brs, 1H), 6.48-6.36 (m, 2H), 4.83 (s, 2H), 4.30 (m, 2H), 3.84 (m, 2H), 2.67 (m, 2H), 2.20 (s, 3H). LC-MS: [M+H]+=339.1.
Compound E-Y6 (39 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Benzoic anhydride ((PhCO)2O, 15 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y21 as a white solid (13 mg, yield in two steps 32%).
1H NMR (400 MHz, MeOD-d4) δ 9.30 (s, 1H), 7.82 (s, 1H), 7.51 (m, 5H), 7.48 (s, 1H), 6.92 (s, 1H), 6.47-6.36 (m, 2H), 4.82 (s, 2H), 4.46 (m, 1H), 4.24 (m, 1H), 4.06 (m, 1H), 3.71 (m, 1H), 2.74 (m, 2H). LC-MS: [M+H]+=401.1
Compound E-Y6 (39 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Methanesulfonic anhydride (Ms2O, 26 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1. Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y22 as a white solid (18 mg, yield in two steps 47%).
1H NMR (400 MHz, DMSO-d6) δ 9.43 (s, 1H), 8.86 (s, 1H), 7.71 (s, 1H), 7.63 (s, 1H), 7.30 (s, 1H), 6.43 (m, 2H), 4.73 (s, 2H), 3.94 (m, 2H), 3.40 (m, 2H), 2.95 (s, 3H), 2.69 (m, 2H). LC-MS: [M+H]+=375.1.
Compound E-Y6 (39 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. p-toluenesulfonic acid anhydride (Ts2O, 48 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y23 as a white solid (16 mg, yield in two steps 35%).
1H NMR (400 MHz, MeOD-d4) δ 9.24 (s, 1H), 7.76 (d, J=7.9 Hz, 2H), 7.67 (s, 1H), 7.47 (s, 1H), 7.46 (d, J=8.4 Hz, 2H), 6.93 (s, 1H), 6.40 (m, 2H), 4.78 (s, 2H), 3.82 (m, 2H), 3.36 (m, 2H), 2.71 (m, 2H), 2.46 (s, 3H). LC-MS: [M+H]+=451.1.
Compound E-Y6 (39 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Ethyl isocyanate (11 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y24 as a white solid (20 mg, yield in two steps 55%).
1H NMR (400 MHz, MeOD-d4) δ 9.25 (s, 1H), 7.70 (s, 1H), 7.47 (d, J=1.0 Hz, 1H), 6.98 (m, 1H), 6.44-6.36 (m, 2H), 4.79 (s, 2H), 4.11 (d, J=2.8 Hz, 2H), 3.69 (t, J=5.6 Hz, 2H), 3.25 (q, J=7.2 Hz, 2H), 2.66 (d, J=16.7 Hz, 2H), 1.16 (t, J=7.2 Hz, 3H). LC-MS: [M+H]+=368.1.
Compound E-Y6 (39 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring, t-butyl isocyanate (11 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y25 as a white solid (14 mg, yield in two steps 37%).
1H NMR (400 MHz, MeOD-d4) δ 9.28 (s, 1H), 7.76 (s, 1H), 7.48 (m, 1H), 6.90 (s, 1H), 6.45-6.34 (m, 2H), 4.80 (s, 2H), 4.10 (d, J=2.5 Hz, 2H), 3.65 (t, J=5.6 Hz, 2H), 2.63 (m, 2H), 1.38 (s, 9H). LC-MS: [M+H]+=396.1.
Compound E-Y6 (39 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring, p-toluene isocyanate (20 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y26 as a white solid (20 mg, yield in two steps 46%).
1H NMR (400 MHz, DMSO-d6) δ 9.44 (s, 1H), 8.81 (m, 1H), 8.46 (s, 1H), 7.72 (s, 1H), 7.63 (s, 1H), 7.37 (d, J=8.4 Hz, 2H), 7.32 (s, 1H), 7.05 (d, J=8.2 Hz, 2H), 6.43 (s, 2H), 4.74 (d, J=4.8 Hz, 2H), 4.22 (s, 2H), 3.69 (t, J=5.4 Hz, 2H), 2.62 (s, 2H), 2.24 (s, 3H). LC-MS: [M+H]+=430.1.
Compound E-Y6 (39 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. p-chlorobenzene isocyanate (23 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1. Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y27 as a white solid (23 mg, yield in two steps 51%).
1H NMR (400 MHz, DMSO-d6) δ 9.44 (s, 1H), 8.81 (s, 1H), 8.70 (s, 1H), 7.72 (s, 1H), 7.63 (s, 1H), 7.54 (d, J=8.9 Hz, 2H), 7.32 (s, 1H), 7.29 (d, J=8.9 Hz, 2H), 6.43 (s, 2H), 4.74 (s, 2H), 4.23 (s, 2H), 3.71 (t, J=5.6 Hz, 2H), 2.63 (s, 2H). LC-MS: [M+H]+=450.1.
Compound E-Y6 (39 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Benzyl isocyanate (20 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y28 as a white solid (20 mg, yield in two steps 47%).
1H NMR (400 MHz, DMSO-d6) δ 9.43 (s, 1H), 8.79 (s, 1H), 7.69 (s, 1H), 7.63 (s, 1H), 7.37-7.25 (m, 5H), 7.21 (t, J=6.6 Hz, 1H), 7.16 (dd, J=14.8, 9.1 Hz, 1H), 6.43 (d, J=1.7 Hz, 2H), 4.74 (s, 2H), 4.29 (d, J=5.8 Hz, 2H), 4.11 (s, 2H), 3.60 (t, J=5.6 Hz, 2H), 2.55 (s, 2H). LC-MS: [M+H]+=430.1.
Compound E-Y6 (39 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Glutaric anhydride (17 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y29 as a white solid (20 mg, yield in two steps 47%).
1H NMR (400 MHz, MeOD-d4) δ 9.32 (s, 1H), 7.89 (d, J=4.5 Hz, 1H), 7.49 (s, 1H), 6.76 (d, J=26.4 Hz, 1H), 6.46-6.34 (m, 2H), 4.83 (s, 2H), 4.31 (m, 2H), 3.92-3.79 (m, 2H), 2.67 (m, 2H), 2.60-2.49 (m, 2H), 2.46-2.36 (m, 2H), 1.95 (m, 2H). LC-MS: [M+H]+=411.1.
Compound E-Y9 (41 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Acetic anhydride (Ac2O, 15 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y30 as a white solid (12 mg, yield in two steps 33%).
1H NMR (400 MHz, MeOD-d4) δ 9.33 (s, 1H), 7.94 (s, 1H), 7.49 (dd, J=1.8, 0.8 Hz, 1H), 6.49 (s, 1H), 6.47-6.37 (m, 2H), 4.84 (s, 2H), 4.06 (m, 1H), 2.62 (m, 3H), 2.32-2.15 (m, 1H), 2.07 (m, 1H), 1.99 (s, 3H), 1.80 (m, 1H). LC-MS: [M+H]+=353.1.
Compound E-Y9 (41 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Benzoic anhydride ((PhCO)2O, 15 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y31 as a white solid (12 mg, yield in two steps 27%).
1H NMR (400 MHz, MeOD-d4) δ 9.29 (s, 1H), 7.90-7.83 (m, 2H), 7.81 (s, 1H), 7.56 (m, 1H), 7.51-7.44 (m, 3H), 6.73 (m, 1H), 6.47-6.37 (m, 2H), 4.81 (s, 2H), 4.30 (m, 1H), 2.73 (m, 3H), 2.21 (m, 1H), 2.05 (m, 1H), 1.97 (m, 1H). LC-MS: [M+H]+=415.1.
Compound E-Y9 (41 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Methanesulfonic anhydride (Ms2O, 26 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y32 as a white solid (11 mg, yield in two steps 29%).
1H NMR (400 MHz, MeOD-d4) δ 9.32 (s, 1H), 7.89 (s, 1H), 7.49 (dd, J=1.8, 0.8 Hz, 1H), 6.51 (m, 1H), 6.45-6.38 (m, 2H), 4.83 (s, 2H), 3.66 (m, 1H), 3.03 (s, 3H), 2.67 (m, 3H), 2.38-2.25 (m, 1H), 2.23-2.14 (m, 1H), 1.94-1.79 (m, 1H). LC-MS: [M+H]+=389.1.
Compound E-Y9 (41 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Glutaric anhydride (17 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y33 as a white solid (16 mg, yield in two steps 38%).
1H NMR (400 MHz, DMSO-d6) δ 12.21 (brs, 1H), 9.41 (s, 1H), 8.75 (s, 1H), 7.85 (d, J=6.9 Hz, 1H), 7.64 (d, J=4.0 Hz, 2H), 7.19 (s, 1H), 6.42 (s, 2H), 4.72 (s, 2H), 3.85 (m, 1H), 2.60 (m, 2H), 2.48-2.33 (m, 2H), 2.21 (m, 2H), 2.11 (m, 2H), 1.91 (m, 1H), 1.73 (m, 2H), 1.61 (m 1H). LC-MS: [M+H]+=425.1.
Compound E-Y6 (39 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) were added and dissolved by stirring. HATU (76 mg, 0.2 mmol), 2,2-difluoropropionic acid (18 mg, 0.16 mmol) were added at room temperature, and the reaction was performed for 6 hours. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y34 as a white solid (10 mg, yield in two steps 25%).
1H NMR (400 MHz, MeOD-d4) δ 9.29 (s, 1H), 7.82 (m, 1H), 7.49 (dd, J=1.8, 0.8 Hz, 1H), 6.91 (m, 1H), 6.50-6.35 (m, 2H), 4.81 (s, 2H), 4.51 (m, 1H), 4.34 (m, 1H), 4.03 (t, J=5.6 Hz, 1H), 3.93 (t, J=5.6 Hz, 1H), 2.76 (m, 2H), 1.87 (td, J=19.9, 6.4 Hz, 3H). LC-MS: [M+H]+=389.1.
Compound E-Y6 (39 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) were added and dissolved by stirring. HATU (76 mg, 0.2 mmol) and N,N-dimethylglycine (17 mg, 0.16 mmol) were added at room temperature, and the reaction was performed for 6 hours. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1. Rf=0.5), the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y35 as a white solid (11 mg, yield in two steps 28%).
1H NMR (400 MHz, MeOD-d4) δ 9.29 (s, 1H), 7.77 (d, J=8.5 Hz, 1H), 7.48 (m, 1H), 6.92 (d, J=30.7 Hz, 1H), 6.47-6.32 (m, 2H), 4.81 (s, 2H), 4.38 (s, 1H), 4.35 (m, 1H), 4.32 (s, 1H), 4.20 (m, 1H), 3.93 (t, J=5.8 Hz, 1H), 3.70 (t, J=5.7 Hz, 1H), 3.00 (d, J=3.4 Hz, 6H). LC-MS: [M+H]+=382.1.
Compound E-Y6 (39 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) were added and dissolved by stirring. HATU (76 mg, 0.2 mmol) and morpholin-4-ylacetic acid (23 mg, 0.16 mmol) were added at room temperature, and the reaction was performed for 6 hours. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y36 as a white solid (9 mg, yield in two steps 21%).
LC-MS: [M+H]+=424.2.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Benzyl isocyanate (20 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y37 as a white solid (12 mg, yield in two steps 23%).
1H NMR (400 MHz, DMSO-d6) δ 9.45 (s, 1H), 8.64 (t, J=4.9 Hz, 1H), 7.69 (s, 1H), 7.30 (dd, J=12.0, 5.0 Hz, 6H), 7.25-7.12 (m, 2H), 7.00-6.91 (m, 1H), 6.70 (dd, J=8.6, 3.8 Hz, 1H), 4.69 (d, J=4.9 Hz, 2H), 4.54 (t, J=8.7 Hz, 2H), 4.28 (d, J=5.6 Hz, 2H), 4.10 (s, 2H), 3.28 (t, J=8.8 Hz, 3H), 2.55 (s, 2H). LC-MS: [M+H]+=500.1.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. p-chlorobenzene isocyanate (23 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y38 as a white solid (18 mg, yield in two steps 34%).
1H NMR (400 MHz, DMSO-d6) δ 9.46 (s, 1H), 8.67 (d, J=21.7 Hz, 2H), 7.72 (s, 1H), 7.54 (d, J=8.1 Hz, 2H), 7.29 (d, J=8.4 Hz, 3H), 6.95 (s, 1H), 6.72 (s, 1H), 4.70 (s, 2H), 4.53 (d, J=8.2 Hz, 2H), 4.23 (s, 2H), 3.70 (s, 2H), 3.29 (s, 2H), 2.63 (s, 2H).
LC-MS: [M+H]+=520.1.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Benzoyl isothiocyanate (24 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y39 as a white solid (10 mg, yield in two steps 19%).
1H NMR (400 MHz, DMSO-d6) δ 10.89 (d, J=17.5 Hz, 1H), 9.47 (d, J=5.7 Hz, 1H), 8.69 (s, 1H), 7.98 (t, J=8.1 Hz, 2H), 7.75 (d, J=25.1 Hz, 1H), 7.63 (s, 1H), 7.53 (d, J=8.1 Hz, 2H), 7.21 (s, 1H), 7.01-6.91 (m, 1H), 6.79-6.63 (m, 1H), 4.76 (s, 1H), 4.70 (d, J=5.0 Hz, 2H), 4.54 (t, J=8.6 Hz, 2H), 4.36 (s, 2H), 3.82 (s, 1H), 3.29 (t, J=8.7 Hz, 3H), 2.79 (s, 2H). LC-MS: [M+H]+=530.1.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) were added and dissolved by stirring. HATU (CAS No.: 148893-10-1) (76 mg, 0.2 mmol) and morpholin-4-ylacetic acid (23 mg, 0.16 mmol) were added at room temperature, and the reaction was performed for 6 hours. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y40 as a white solid (5 mg, yield in two steps 10%).
1H NMR (400 MHz, DMSO-d6) δ 9.48 (d, J=2.5 Hz, 1H), 8.72 (s, 1H), 7.73 (d, J=9.0 Hz, 1H), 7.31 (d, J=17.2 Hz, 1H), 6.95 (t, J=9.4 Hz, 1H), 6.71 (dd, J=8.6, 3.7 Hz, 1H), 4.70 (m, 2H), 4.54 (t, J=8.4 Hz, 2H), 4.44 (d, J=25.8 Hz, 2H), 4.23 (d, J=30.0 Hz, 2H), 3.96 (m, 2H), 3.78 (m, 2H), 3.61 (m, 2H), 3.44 (m, 2H), 3.29 (m, 2H), 3.17 (m, 2H), 2.67 (m, 2H). LC-MS: [M+H]+=494.2.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) were added and dissolved by stirring. HATU (76 mg, 0.2 mmol) and oxazole-5-carboxylic acid (18 mg, 0.16 mmol) were added at room temperature, and the reaction was performed for 6 hours. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y41 as a white solid (10 mg, yield in two steps 22%).
1H NMR (400 MHz, MeOD-d4) δ 9.32 (s, 1H), 8.41 (s, 1H), 7.81 (s, 1H), 7.77 (s, 1H), 6.95 (s, 1H), 6.91-6.81 (m, 1H), 6.66 (dd, J=8.6, 3.9 Hz, 1H), 4.80 (s, 2H), 4.59 (t, J=8.7 Hz, 3H), 4.43 (m, 1H), 4.05 (m, 2H), 3.38 (t, J=8.7 Hz, 2H), 2.93-2.65 (m, 2H). LC-MS: [M+H]+=462.2.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) were added and dissolved by stirring. HATU (76 mg, 0.2 mmol) and 1,3-dimethyl-1H-pyrazole-5-carboxylic acid (22 mg, 0.16 mmol) were added at room temperature, and the reaction was performed for 6 hours. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y42 as a white solid (10 mg, yield in two steps 19%).
1H NMR (400 MHz, MeOD-d4) δ 9.40 (s, 1H), 7.98 (s, 1H), 6.95-6.80 (m, 1H), 6.76-6.52 (m, 2H), 6.37 (s, 1H), 4.82 (s, 2H), 4.59 (t, J=8.7 Hz, 2H), 4.40 (d, J=24.2 Hz, 2H), 4.01 (d, J=10.7 Hz, 1H), 3.87 (s, 4H), 3.39 (t, J=8.5 Hz, 2H), 2.72 (s, 2H), 2.28 (s, 3H). LC-MS: [M+H]+=489.2.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) were added and dissolved by stirring. HATU (76 mg, 0.2 mmol) and tetrahydropyran-4-carboxylic acid (21 mg, 0.16 mmol) were added at room temperature, and the reaction was performed for 6 hours. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y43 as a white solid (10 mg, yield in two steps 21%).
1H NMR (400 MHz, MeOD-d4) δ 9.36 (s, 1H), 7.86 (d, J=5.7 Hz, 1H), 6.91-6.58 (m, 3H), 4.79 m, 2H), 4.58 (m, 2H), 4.38 (s, 1H), 4.27 (s, 1H), 3.99 (m, 2H), 3.87 (m, 2H), 3.55 (m, 2H), 3.37 (m, 2H), 3.04 (m, 1H), 2.65 (m, 2H), 1.83 (m, 2H), 1.68 (m, 2H). LC-MS: [M+H]+=479.2.
Bromide 4-1 (72 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-7 (137 mg, 0.4 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y44 as a white solid (18 mg, 36%).
1H NMR (400 MHz, DMSO-d6) δ 9.46 (s, 1H), 8.67 (s, 1H), 7.69 (s, 1H), 7.38 (dd, J=7.5, 4.3 Hz, 6H), 7.24 (s, 1H), 7.03-6.88 (m, 1H), 6.70 (dd, J=8.7, 3.9 Hz, 1H), 5.12 (d, J=12.1 Hz, 2H), 4.68 (s, 2H), 4.61-4.46 (m, 2H), 4.17 (m, 2H), 3.65 (m, 2H), 3.28 (t, J=8.7 Hz, 2H), 2.58 (m, 2H). LC-MS: [M+H]+=501.1.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Acrylic anhydride (20 mg, 0.15 mmol) was added at room temperature, and the reaction was further performed for 1 hour. When the reaction was completed as indicated by TLC (DCM:MeOH=10:1, Rf=0.5), the resultant was directly separated by column chromatography (DCM:MeOH=30:1) to obtain the target compound E-Y45 as a white solid (14 mg, yield in two steps 35%).
1H NMR (400 MHz, MeOH-d4) δ 9.38 (s, 1H), 7.92 (s, 1H), 6.94-6.80 (m, 2H), 6.79-6.68 (m, 1H), 6.65 (dd, J=8.6, 3.9 Hz, 1H), 6.27 (d, J=16.7 Hz, 1H), 5.81 (d, J=10.6 Hz, 1H), 4.81 (s, 2H), 4.59 (t, J=8.7 Hz, 2H), 4.37 (m, 2H), 3.92 (m, 2H), 3.38 (m, 2H), 2.68 (m, 2H). LC-MS: [M+H]+=421.1.
Bromide 4-3 (62 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-3 (123 mg, 0.4 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y46 as a white solid (32 mg, 40%).
1H NMR (400 MHz, MeOD-d4) δ 9.43 (s, 1H), 7.71 (s, 1H), 7.47 (m, 1H), 7.16 (m, 1H), 6.99 (m, 2H), 4.91 (s, 2H), 4.08 (m, 2H), 3.57 (m, 2H), 2.78 (m, 2H), 1.44 (s, 9H). LC-MS: [M+H]+=412.1.
Compound E-Y1 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (5 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography (DCM:MeOH=20:1) to obtain the target compound E-Y47 as a white solid (8 mg, 80%).
1H NMR (400 MHz, MeOD-d4) δ 9.33 (s, 1H), 7.74 (s, 1H) , 6.93-6.80 (m, 1H), 6.66 (dd, J=8.7, 3.8 Hz, 1H), 4.78 (s, 2H), 4.59 (t, J=8.7 Hz, 2H), 4.09 (d, J=11.2 Hz, 2H), 3.71-3.57 (m, 2H), 3.38 (t, J=8.7 Hz, 2H), 3.21 (m, 1H), 1.96 (m, 4H). LC-MS: [M+H]+=370.1.
Compound E-Y17 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (5 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography (DCM:MeOH=20:1) to obtain the target compound E-Y48 as a white solid (5 mg, 50%).
1H NMR (400 MHz, MeOD-d4) δ 9.27 (s, 1H), 7.58 (s, 1H), 6.99-6.77 (m, 1H), 6.66 (dd, J=8.5, 3.7 Hz, 1H), 4.76 (s, 2H), 4.59 (t, J=8.7 Hz, 2H), 4.20 (m, 2H), 3.37 (m, 2H), 3.23 (dd, J=14.2, 7.1 Hz, 2H), 2.96 (t, J=11.7 Hz, 2H), 2.04 (m, 4H), 1.15 (t, J=7.2 Hz, 3H). LC-MS: [M+H]+=440.2.
Compound E-Y18 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (5 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography (DCM:MeOH=20:1) to obtain the target compound E-Y49 as a white solid (6 mg, 60%).
1H NMR (400 MHz, DMSO-d6) δ 9.40 (s, 1H), 8.44 (s, 1H), 7.49 (s, 1H), 7.02-6.87 (m, 1H), 6.70 (dd, J=8.6, 3.9 Hz, 1H), 4.64 (d, J=4.8 Hz, 2H), 4.54 (t, J=8.8 Hz, 2H), 4.10 (d, J=12.9 Hz, 2H), 3.29 (t, J=8.7 Hz, 2H), 2.96 (s, 1H), 2.77-2.64 (m, 2H), 1.90-1.68 (m, 4H), 1.27 (s, 9H). LC-MS: [M+H]+=468.2.
Compound E-Y15 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (5 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography (DCM:MeOH=20:1) to obtain the target compound E-Y50 as a white solid (7 mg, 72%).
1H NMR (400 MHz, MeOD) δ 9.28 (s, 1H), 7.63 (s, 1H), 6.92-6.78 (m, 1H), 6.66 (dd, J=8.7, 3.9 Hz, 1H), 4.77 (s, 2H), 4.71 (m, 1H), 4.59 (t, J=8.7 Hz, 2H), 4.09 (m, 1H), 3.37 (t, J=8.6 Hz, 2H), 3.30-3.14 (m, 1H), 2.80 (m, 1H), 2.25-2.18 (m, 1H), 2.17 (s, 3H), 2.06 (m, 2H), 1.91 (m, 1H), 1.81 (m, 1H). LC-MS: [M+H]+=411.2.
Bromide 13-2 (63 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-3 (123 mg, 0.4 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y51 as a white solid (17 mg, 20% o).
1H NMR (400 MHz, MeOD-d4) δ 9.48 (s, 1H), 7.50 (s, 1H), 7.34 (s, 1H), 6.95 (s, 1H), 6.47 (m, 2H), 5.08 (s, 2H), 4.18 (m, 2H), 3.72 (m, 2H), 2.62 (m, 2H), 1.48 (s, 9H). LC-MS: [M+H]+=421.2.
Bromide 13-1 (77 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-1 (84 mg, 0.4 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y52 as a white solid (20 mg, 21%). LC-MS: [M+H]+=392.2.
Bromide 13-1 (77 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-3 (123 mg, 0.4 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound E-Y53 as a white solid (13 mg, 14%). LC-MS: [M+H]+=491.2.
Compound E-Y3 (46 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) were added and dissolved by stirring. HATU (76 mg, 0.2 mmol) and N,N-dimethylglycine (17 mg, 0.16 mmol) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound E-Y54 (6 mg).
1H NMR (400 MHz, DMSO-d6) δ 9.54 (d, J=3.3 Hz, 1H), 8.79 (s, 1H), 7.70 (d, J=6.1 Hz, 1H), 7.30 (s, 1H), 6.95 (t, J=9.4 Hz, 1H), 6.70 (dd, J=8.5, 3.7 Hz, 1H), 4.69 (d, J=4.6 Hz, 2H), 4.53 (t, J=8.7 Hz, 2H), 4.24 (d, J=41.1 Hz, 2H), 3.71 (s, 2H), 3.48 (s, 2H), 3.29 (t, J=8.7 Hz, 2H), 2.60 (d, J=38.4 Hz, 2H), 2.37 (s, 6H). LC-MS: [M+H]+=452.2.
Bromide 4-1 (72 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-8 (89 mg, 0.4 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound SL-ZYE-07 (47 mg, 62%). 1H NMR (400 MHz, DMSO-d6) δ 9.39 (s, 1H), 8.51 (s, 1H), 7.56 (s, 1H), 6.91 (t, J=9.3 Hz, 1H), 6.69 (m, 2H), 4.65 (d, J=3.8 Hz, 2H), 4.51 (t, J=8.5 Hz, 2H), 3.71 (m, 2H), 3.62 (m, 2H), 3.26 (m, 2H), 2.79 (m, 2H), 2.45 (m, 2H). LC-MS: [M+H]+=382.2.
Compound SL-ZYE-07 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (5 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography (DCM:MeOH=20:1) to obtain the target compound SL-ZYE-08 (8 mg, 82%).
1H NMR (400 MHz, CDCl3+MeOD-d4) δ 9.05 (s, 1H), 7.41 (s, 1H), 6.82-6.64 (m, 1H), 6.56 (dd, J=8.6, 3.9 Hz, 1H), 4.60 (s, 2H), 4.52 (t, J=8.7 Hz, 2H), 3.91-3.77 (m, 2H), 3.69 (ddd, J=19.0, 10.7, 6.3 Hz, 2H), 3.27 (t, J=8.7 Hz, 2H), 3.16 (d, J=7.8 Hz, 1H), 2.05-1.91 (m, 4H), 1.84 (dd, J=9.0, 5.1 Hz, 2H). LC-MS: [M+H]+=384.2.
SL-ZYE-08 was further separated by chiral chromatography to obtain a pair of optically pure compounds SL-ZYE-08-S and SL-ZYE-08-R. The separation conditions were: (chiral column: Chiralcel OD-3, 4.6 mm×250 mm, particle size: 3 μm, mobile phase: n-hexane:isopropanol=50:50, flow rate=1 ml/min, two fractions were obtained, fraction 1 (Rt=17.085 min); fraction 2 (Rt=18.627 min). The chiral chromatography separation method was a conventional method known to those skilled in the art.
E-Y2 (5 mg) was dissolved in 1 mL of dichloromethane, mCPBA (m-chloroperoxybenzoic acid) (2 mg) was added thereto and the reaction system was stirred at room temperature for 120 min under Ar atmosphere protection. After being concentrated under reduced pressure, the resultant was separated by column chromatography to obtain the target compound SL-ZYE-09 (2 mg).
1H NMR (400 MHz, DMSO-d6) δ 9.46 (s, 1H), 8.70 (t, J=5.1 Hz, 1H), 7.76 (s, 1H), 7.12 (t, J=4.6 Hz, 1H), 7.01-6.85 (m, 1H), 6.70 (dd, J=8.7, 3.9 Hz, 1H), 4.69 (d, J=4.9 Hz, 2H), 4.54 (t, J=8.7 Hz, 2H), 3.99 (s, 2H), 3.37 (t, J=6.2 Hz, 2H), 3.28 (t, J=8.7 Hz, 2H), 3.17 (d, J=5.3 Hz, 2H). LC-MS: [M+H]+=416.1.
Bromide 4-4 (69 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-1 (84 mg, 0.4 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound SL-ZYE-11 (17 mg).
1H NMR (400 MHz, CDCl3+MeOD-d4) δ 9.08 (s, 1H), 7.56 (s, 1H), 7.14 (s, 1H), 7.02 (d, J=7.4 Hz, 1H), 6.79 (d, J=8.0 Hz, 1H), 6.66 (d, J=7.9 Hz, 1H), 4.53 (t, J=8.7 Hz, 2H), 4.35 (s, 2H), 3.92 (t, J=5.3 Hz, 2H), 3.30 (s, 2H), 3.19 (t, J=8.4 Hz, 2H), 2.53 (s, 2H). LC-MS: [M+H]J=350.2.
Compound SL-ZYE-11 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (5 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography (DCM:MeOH=20:1) to obtain the target compound SL-ZYE-14 (5 mg).
1H NMR (400 MHz, DMSO-d6) δ 9.41 (s, 1H), 8.55 (t, J=5.4 Hz, 1H), 7.45 (s, 1H), 7.06 (t, J=7.8 Hz, 1H), 6.85 (d, J=7.5 Hz, 1H), 6.69 (d, J=7.9 Hz, 1H), 4.63 (d, J=5.3 Hz, 2H), 4.54 (t, J=8.7 Hz, 2H), 3.96 (dd, J=10.8, 3.6 Hz, 2H), 3.51-3.44 (m, 2H), 3.22 (t, J=8.7 Hz, 2H), 3.09 (ddd, J=15.4, 7.9, 3.7 Hz, 1H), 2.01-1.87 (m, 2H), 1.82 (d, J=12.5 Hz, 2H). LC-MS: [M+H]+=352.2.
Bromide 4-5 (68 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-1 (84 mg, 0.4 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound SL-ZYE-17 (17 mg).
1H NMR (400 MHz, MeOD-d4) δ 9.26 (s, 1H), 7.78 (d, J=2.3 Hz, 1H), 7.74 (s, 1H), 7.47 (d, J=7.9 Hz, 1H), 7.30 (d, J=7.6 Hz, 2H), 7.08 (s, 1H), 7.03 (s, 1H), 5.07 (s, 2H), 4.39 (d, J=2.6 Hz, 2H), 3.98 (t, J=5.5 Hz, 2H), 2.63 (s, 2H). LC-MS: [M+H]+=348.1.
Bromide 4-5 (68 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-3 (123 mg, 0.4 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound SL-ZYE-18 (15 mg).
1H NMR (400 MHz, CDCl3) δ 8.77 (s, 1H), 7.71 (s, 1H), 7.67 (d, J=2.3 Hz, 1H), 7.51 (s, 1H), 7.31-7.29 (m, 4H), 6.90 (s, 1H), 5.08 (d, J=5.3 Hz, 2H), 4.18 (s, 2H), 3.70 (s, 2H), 2.64 (s, 2H), 1.51 (s, 9H). LC-MS: [M+H]+=447.1.
Compound E-Y20 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (3 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography (DCM:MeOH=20:1) to obtain the target compound E-Y20-H (4 mg, 40%).
1H NMR (400 MHz, MeOD-d4) δ 9.23 (s, 1H), 7.57 (d, J=0.7 Hz, 1H), 7.47 (dd, J=1.8, 0.9 Hz, 1H), 6.42-6.36 (m, 2H), 4.77 (s, 2H), 4.73 (m, 1H), 4.09 (m, 1H), 3.35-3.17 (m, 2H), 2.80 (dd, J=12.9, 10.2 Hz, 1H), 2.24-2.18 (m, 1H), 2.17 (s, 3H), 2.07 (m, 1H), 1.87 (m, 2H). LC-MS: [M+H]+=341.1.
Compound E-Y13 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (3 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography (DCM:MeOH=20:1) to obtain the target compound E-Y13-H (7 mg, 71%).
1H NMR (400 MHz, DMSO-d6) δ 9.41 (s, 1H), 8.48 (t, J=5.2 Hz, 1H), 7.50 (s, 1H), 6.98-6.90 (m, 1H), 6.70 (dd, J=8.6, 3.9 Hz, 1H), 4.64 (d, J=5.0 Hz, 2H), 4.54 (t, J=8.7 Hz, 2H), 3.69 (d, J=11.6 Hz, 2H), 3.29 (t, J=8.7 Hz, 2H), 3.01 (m, 1H), 2.91 (s, 3H), 2.85 (dd, J=8.7, 6.0 Hz, 2H), 2.07-1.85 (m, 4H). LC-MS: [M+H]+=447.2.
Compound E-Y (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (3 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and the solvent was dried by rotatory evaporator to obtain the target compound E-Y2-H LC-MS: [M+H]+=386.1.
5 mg of E-Y2-H was dissolved in 1 mL of dichloromethane solution, mCPBA (5 mg) was added to the solution, and the reaction was performed at room temperature for 3 h, and the resultant was separated by column chromatography to obtain the target compound SL-ZYE-34 (1.5 mg)
1H NMR (400 MHz, MeOD-d4) δ 9.27 (s, 1H), 7.63 (s, 1H), 6.92-6.78 (m, 1H), 6.65 (dd, J=8.6, 3.7 Hz, 1H), 4.76 (s, 2H), 4.59 (t, J=8.7 Hz, 2H), 3.45-3.33 (m, 5H), 3.17 (d, J=11.8 Hz, 2H), 2.55-2.33 (m, 4H). LC-MS: [M+H]+=418.2.
Bromide 4-1 (72 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-10 (101 mg, 0.4 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (11) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound SL-ZYE-23 (14 mg).
1H NMR (400 MHz, DMSO-d6) δ 9.46 (s, 1H), 8.67 (s, 1H), 7.72 (s, 1H), 7.27 (s, 1H), 7.00-6.87 (m, 1H), 6.70 (dd, J=8.7, 3.8 Hz, 2H), 4.70 (d, J=4.8 Hz, 2H), 4.54 (t, J=8.6 Hz, 2H), 4.30 (d, J=5.7 Hz, 2H), 3.27 (t, J=8.7 Hz, 2H), 2.68 (s, 2H), 1.45 (s, 9H), 1.29 (d, J=6.4 Hz, 3H), 1.09 (d, J=6.4 Hz, 3H). LC-MS: [M+H]+=495.3.
Bromide 4-1 (72 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-1 (95 mg, 0.4 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound SL-ZYE-24 (21 mg).
1H NMR (400 MHz, CDCl3) δ 8.93 (s, 1H), 7.66 (s, 1H), 7.24 (s, 1H), 6.88-6.70 (m, 1H), 6.61 (dd, J=8.7, 4.0 Hz, 1H), 6.35 (s, 1H), 4.79 (d, J=5.5 Hz, 2H), 4.61 (t, J=8.7 Hz, 2H), 4.51 (s, 1H), 3.94-3.82 (m, 1H), 3.50 (m, 1H), 3.38 (t, J=8.8 Hz, 2H), 2.45 (t, J=16.4 Hz, 2H), 1.38 (d, J=6.2 Hz, 3H), 1.35 (d, J=6.8 Hz, 3H). LC-MS: [M+H]+=396.2.
Bromide 4-1 (72 mg, 0.2 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-12 (103 mg, 0.4 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (7.3 mg, 0.02 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (21.2 mg, 0.04 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography (DCM:MeOH=20:1), to obtain the target compound SL-ZYE-28 (27 mg, 32%).
1H NMR (400 MHz, MeOD-d4) δ 9.44 (s, 1H), 8.07 (s, 1H), 7.23-7.14 (m, 1H), 6.94-6.79 (m, 4H), 6.68 (dd, J=8.6, 3.9 Hz, 1H), 6.17 (t, J=3.9 Hz, 1H), 4.92 (d, J=3.9 Hz, 2H), 4.88 (s, 2H), 4.61 (t, J=8.7 Hz, 2H), 3.43 (t, J=8.7 Hz, 2H). LC-MS: [M+H]+=416.2.
Compound E-Y54 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (5 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography (DCM:MeOH=20:1) to obtain the target compound E-Y54-H (5 mg).
1H NMR (400 MHz, MeOD-d4) δ 9.27 (s, 1H), 7.57 (s, 1H), 6.91-6.81 (m, 1H), 6.65 (dd, J=8.6, 3.9 Hz, 1H), 4.76 (s, 2H), 4.70 (d, J=13.5 Hz, 1H), 4.58 (t, J=8.7 Hz, 2H), 4.10 (d, J=13.9 Hz, 1H), 3.60 (dd, J=16.9, 6.6 Hz, 2H), 3.36 (dd, J=10.0, 7.6 Hz, 2H), 3.25 (d, J=16.0 Hz, 2H), 2.84 (m, 1H), 2.52 (s, 6H), 2.07 (m, 2H), 1.91 (m, 2H). LC-MS: [M+H]+=454.2.
Bromide 4-1 (36 mg, 0.1 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-13 (64 mg, 0.2 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (3.5 mg, 0.01 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (11 mg, 0.02 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography to obtain the target compound SL-E1 (11 mg). LC-MS: [M+H]+=481.2.
Compound SL-E1 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (5 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E2 (7 mg). LC-MS: [M+H]+=483.2.
Referring to Example 56, SL-E2 was separated by a chiral chromatography column to obtain optically pure compounds SL-E2-S and SL-E2-R.
Compound SL-E3 (9 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E3 (5 mg). LC-MS: [M+H]+=497.2.
Compound SL-E3 (9 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E4 (5 mg). LC-MS: [M+H]+=398.1.
Compound E-Y10 (20 mg) was dissolved in 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg) and 2-(1-pyrrolidinyl) acetic acid (CAS: 37386-15-5) (13 mg) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E5 (3 mg). LC-MS: [M+H]+=478.2.
Compound E-Y10 (20 mg) was dissolved in 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg, 0.1 mmol) and N. N-diethylglycine (13 mg, 0.1 mmol) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E6 (6 mg). LC-MS: [M+H]+=480.2.
Compound E-Y10 (20 mg) was dissolved in 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg) and methoxyacetic acid (CAS: 625-45-6) (9 mg) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E7 (2 mg). LC-MS: [M+H]+=439.2.
Compound E-Y10 (20 mg) was dissolved in 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg, 0.1 mmol) and acid (oxetane-3-carboxylic acid) (10 mg, 0.1 mmol) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E8 (2.5 mg). LC-MS: [M+H]+=451.2.
Compound E-Y10 (20 mg) was dissolved in 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg) and 2-(2-methoxyethoxy) acetic acid (CAS No.: 16024-56-9) (14 mg) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E9 (5 mg). LC-MS: [M+H]+=483.2.
Compound E-Y10 (20 mg) was dissolved in 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg, 0.1 mmol) and 1-methylpiperidine-4-carboxylic acid (15 mg, 0.1 mmol) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E10 (6 mg). MS: [M+H]+=492.2.
Compound E-Y10 (20 mg) was dissolved in 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg, 0.1 mmol) and acid (CAS No.: 1158712-36-7) (22 mg, 0.1 mmol) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E11 (7 mg). LC-MS: [M+H]+=562.2.
The compound SL-E1 (96 mg, 0.2 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After being concentrated, the resultant was directly separated by HPLC to obtain the product SL-E12 (50 mg), LC-MS: [M+H]+=381.2.
SL-E12 (25 mg) was dissolved in 1 mL of dichloromethane and 0.1 mL of triethylamine (TEA) by stirring. Acetic anhydride (Ac2O, 15 mg) was added at room temperature, and the reaction was performed for 1 hour. After the reaction was completed as indicated by TLC, the resultant was separated by column chromatography to obtain the target compound SL-E13 (6 mg). LC-MS: [M+H]+=423.2.
SL-E12 (10 mg) was dissolved in 1 mL of dichloromethane and 0.1 mL of triethylamine (TEA) by stirring. Methanesulfonic anhydride (Ms2O, 8 mg) was added at room temperature, and the reaction was performed for 1 hour. After the reaction was completed as indicated by TLC, the resultant was separated by column chromatography to obtain the target compound SL-E14 (3 mg). 1H NMR (400 MHz, DMSO-d6) δ 9.44 (s, 1H), 8.63 (m, 1H), 7.67 (s, 1H), 6.99-6.90 (m, 1H), 6.82 (t, J=5.9 Hz, 1H), 6.70 (dd, J=8.7, 3.8 Hz, 1H), 4.68 (d, J=4.9 Hz, 2H), 4.54 (t, J=8.7 Hz, 2H), 4.03 (d, J=6.0 Hz, 2H), 3.57-3.48 (m, 2H), 3.31-3.24 (m, 2H), 2.91 (s, 3H), 2.83 (m, 2H), 1.91 (m, 2H). LC-MS: [M+H]+=459.2.
Compound SL-E12 (19 mg, 0.05 mmol) was dissolved in 1 mL of DCM and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg, 0.1 mmol) and N, N-dimethylglycine (11 mg, 0.1 mmol) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E15 (2 mg). LC-MS: [M+H]+=466.3.
Compound SL-E12 (19 mg, 0.05 mmol) was dissolved in 1 mL of DCM and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg, 0.1 mmol) and 2-(1-pyrrolidinyl) acetic acid (CAS: 37386-15-5) (14 mg) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E16 (2 mg). LC-MS: [M+H]+=492.2.
Compound SL-E12 (19 mg, 0.05 mmol) was dissolved in 1 mL of DCM and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg, 0.1 mmol) and N, N-diethylglycine (13 mg) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E17 (2 mg). LC-MS: [M+H]+=494.4.
Compound SL-E12 (19 mg, 0.05 mmol) was dissolved in 1 mL of DCM and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg) and methoxyacetic acid (CAS: 625-45-6) (11 mg) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E18 (3 mg). LC-MS: [M+H]+=453.2.
Compound SL-E12 (19 mg, 0.05 mmol) was dissolved in 1 mL of DCM and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg, 0.1 mmol) and acid (oxetane-3-carboxylic acid) (10 mg, 0.1 mmol) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E19 (1.5 mg). LC-MS: [M+H]+=465.2.
Compound SL-E12 (19 mg, 0.05 mmol) was dissolved in 1 mL of DCM and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg) and 2-(2-methoxyethoxy) acetic acid (CAS No.: 16024-56-9) (15 mg) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E20 (2 mg). LC-MS: [M+H]+=497.2.
Compound SL-E12 (19 mg, 0.05 mmol) was dissolved in 1 mL of DCM and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg, 0.1 mmol) and 1-methylpiperidine-4-carboxylic acid (15 mg, 0.1 mmol) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E21 (3 mg). LC-MS: [M+H1]=506.2.
Compound SL-E12 (19 mg, 0.05 mmol) was dissolved in 1 mL of DCM and 0.1 mL of diisopropylethylamine (DIEPA) by stirring. HATU (38 mg, 0.1 mmol) and acid (CAS No.: 1158712-36-7) (22 mg, 0.1 mmol) were added at room temperature, and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E22 (0.8 mg). LC-MS: [M+H]+=576.2.
Bromide 4-1 (36 mg, 0.1 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-14 (44 mg, 0.2 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (3.5 mg, 0.01 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (11 mg, 0.02 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography, to obtain the target compound SL-E23 as a white solid (6 mg). 1H NMR (400 MHz, DMSO-d6) δ 9.47 (d, J=6.2 Hz, 11H), 8.67 (d, J=4.9 Hz, 1H), 7.71 (d, J=8.4 Hz, 1H), 7.30 (m, 1H), 6.94 (t, J=9.4 Hz, 1H), 6.70 (dd, J=8.5, 3.8 Hz, 1H), 4.69 (d, J=4.8 Hz, 2H), 4.57 (m, 2H), 3.47 (m, 2H), 3.28 (t, J=8.8 Hz, 2H), 2.97 (m, 2H), 2.69 (m, 2H), 2.56 (m, 3H). LC-MS: [M+H]+=381.2.
Bromide 4-1 (36 mg, 0.1 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 aqueous solution, and borate B-15 (47 mg, 0.2 mmol)) was added thereto and the reaction system was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (3.5 mg, 0.01 mmol), 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (11 mg, 0.02 mmol) were added, and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography, to obtain the target compound SL-E24 (3.5 mg). 1H NMR (400 MHz, CD3OD+CDCl3) δ 9.31 (s, 1H), 7.94 (s, 1H), 7.18 (s, 1H), 6.83 (m, 1H), 6.65 (m, 1H), 4.79 (s, 2H), 4.60 (t, J=8.8 Hz, 2H), 3.64 (t, J=7.3 Hz, 2H), 3.37 (m, 2H), 3.06 (s, 3H), 2.99 (m, 2H). LC-MS: [M+H]+=395.1.
Compound SL-E23 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E25 (5 mg). 1H NMR (400 MHz, CD3OD) δ 9.29 (s, 1H), 7.61 (s, 1H), 6.90-6.80 (m, 1H), 6.63 (m, 1H), 4.76 (s, 2H), 4.66-4.51 (m, 2H), 3.58-3.44 (m, 2H), 3.40-3.34 (m, 2H), 3.23-3.11 (m, 1H), 3.09-2.95 (m, 2H), 2.83 (s, 3H), 2.27-2.17 (m, 4H). LC-MS: [M+H]+=383.2.
Compound SL-E24 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E26 (2 mg). 1H NMR (400 MHz, CDCl3) δ 8.94 (s, 1H), 7.45 (s, 1H), 6.84 (t, J=9.4 Hz, 1H), 6.66 (dd, J=8.6, 3.9 Hz, 1H), 6.37 (m, 1H), 4.77 (d, J=5.3 Hz, 2H), 4.63 (t, J=8.7 Hz, 2H), 3.49 (m, 2H), 3.39 (m, 3H), 2.99 (s, 3H), 2.75 (m, 2H), 2.33 (m, 2H). LC-MS: [M+H]+=397.2.
Compound SL-ZYE-23 (50 mg, 0.1 mmol) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After being concentrated, the resultant was directly separated by HPLC to obtain the product SL-E29 (38 mg), LC-MS: [M+H]+=395.2.
SL-E29 (10 mg) was dissolved in 1 mL of dichloromethane and 0.1 mL of triethylamine (TEA) by stirring. Acetic anhydride (Ac2O, 8 mg) was added at room temperature, and the reaction was performed for 1 hour. After the reaction was completed as indicated by TLC, the resultant was separated by column chromatography to obtain the target compound SL-E30 (4 mg). LC-MS: [M+H]+=437.2.
SL-E29 (10 mg) was dissolved in 1 mL of dichloromethane and 0.1 mL of triethylamine (TEA) by stirring. Methanesulfonic anhydride (Ms2O, 7 mg) was added at room temperature, and the reaction was performed for 1 hour. After the reaction was completed as indicated by TLC, the resultant was separated by column chromatography to obtain the target compound SL-E31 (3 mg). LC-MS: [M+H]+=473.2.
SL-E29 (12 mg) was dissolved in 1 mL of dichloromethane and 0.1 mL of triethylamine (TEA) by stirring. HATU (30 mg) and N,N-dimethylglycine (9 mg) were added at room temperature, and the reaction was performed for 1 hour. After the reaction was completed as indicated by TLC, the reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E32 (4 mg). LC-MS: [M+H]+=480.2.
Compound SL-E30 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E33 (4 mg). LC-MS: [M+H]+=439.2.
Compound SL-E31 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added, and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E34 (4 mg). LC-MS: [M+H]+=475.2.
Compound SL-E32 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E35 (4 mg). LC-MS: [M+H]+=482.2.
Compound SL-E5 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E36 (5 mg). LC-MS: [M+H]3=480.2.
Compound SL-E6 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E37 (5 mg). LC-MS: [M+H]+=482.2.
Compound SL-E7 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E38 (5 mg). LC-MS: [M+H]+=441.2.
Compound E-Y3 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 3 hour. The solution was filtered through celite. After the filtrate was dried by rotatory evaporation, the resultant was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the reaction system was stirred at room temperature for 30 min. After the solvent was concentrated, the residue was further dissolved in 1 mL of DMF and 0.1 mL of diisopropylethylamine (DIEPA), and HATU (15 mg) and acid (oxetane-3-carboxylic acid) (9 mg) were added at room temperature and the reaction was performed for 1 hour. The reaction solution was poured into water, and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E39 (0.7 mg). LC-MS: [M+H]+=453.4.
Compound SL-E9 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E40 (5 mg). LC-MS: [M+H]+=485.2.
SL-E10 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E41 (5 mg). LC-MS: [M+H]+=494.3
SL-E11 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E42 (5 mg). LC-MS: [M+H]+=564.2.
SL-E13 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E43 (5 mg) LC-MS: [M+H]+=425.2.
Referring to example 56, SL-E43 was separated by chiral chromatography to obtain optically pure compounds SL-E43-S and SL-E43-R.
SL-E14 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E44 (5 mg). LC-MS: [M+H]+=461.2.
Referring to example 56, SL-E44 was separated by chiral chromatography to obtain optically pure compounds SL-E44-S and SL-E44-R.
SL-E15 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E45 (5 mg). LC-MS: [M+H]+=468.2.
Referring to example 56, SL-E45 was separated by chiral chromatography to obtain optically pure compounds SL-E45-S and SL-E45-R.
SL-E16 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E46 (5 mg). LC-MS: [M+H]+=494.2.
Referring to example 56, SL-E46 was separated by chiral chromatography to obtain optically pure compounds SL-E46-S and SL-E46-R.
SL-E17 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E47 (4 mg). LC-MS: [M+H]+=496.2.
Referring to example 56, SL-E47 was separated by chiral chromatography to obtain optically pure compounds SL-E47-S and SL-E47-R.
SL-E18 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E48 (4 mg) LC-MS: [M+H]+=455.2.
Referring to example 56, SL-E48 was separated by chiral chromatography to obtain optically pure compounds SL-E48-S and SL-E48-R.
Compound SL-E2 (10 mg) was dissolved in 3 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature for 30 min. After the solvent was concentrated, the residue was dissolved in 1 mL of DMF and 0.1 mL of isopropylethylamine (DIEPA), and HATU (30 mg) and acid (oxetane-3-carboxylic acid) (9 mg) were added at room temperature and the reaction was performed for 1 hour. When the reaction was completed as indicated by TLC, the reaction solution was poured into water and extracted with a large amount of ethyl acetate. After being dried and concentrated, the resultant was separated by column chromatography to obtain the target compound SL-E49 (1.2 mg). LC-MS: [M+H]+=467.2.
Referring to example 56, SL-E49 was separated by chiral chromatography to obtain optically pure compounds SL-E49-S and SL-E49-R.
SL-E20 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E50 (4 mg). LC-MS: [M+H]+=499.2.
Referring to example 56, SL-E50 was separated by chiral chromatography to obtain optically pure compounds SL-E50-S and SL-E50-R.
SL-E21 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E51 (4 mg). LC-MS: [M+H]+=508.3.
Referring to example 56, SL-E51 was separated by chiral chromatography to obtain optically pure compounds SL-E51-S and SL-E51-R.
SL-E22 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-E52 (4 mg). LC-MS: [M+H]+=578.2.
Referring to example 56, SL-E52 was separated by chiral chromatography to obtain optically pure compounds SL-E52-S and SL-E52-R.
SL-ZYE-28 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatograph to obtain the target compound SL-E53 (4 mg). LC-MS: [M+H]+=418.2.
Bromide 4-1 (36 mg, 0.1 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 solution, and borate SL-B3 (48 mg, 0.2 mmol) was added thereto. The reaction solution was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (3.5 mg, 0.01 mmol) and 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (11 mg, 0.02 mmol) were added and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography to obtain the target compound SL-ZYE-119. (4.1 mg). 1H NMR (400 MHz, DMSO-d6) δ 9.44 (s, 1H), 8.61 (m, 1H), 7.66 (s, 1H), 7.30 (m, 1H), 6.94 (m, 1H), 6.70 (m, 1H), 4.69 (m, 2H), 4.54 (m, 2H), 4.30 (m, 2H), 3.29 (m, 2H), 2.40 (m, 2H), 1.23 (s, 6H). LC-MS: [M+H]+=396.2.
Bromide 4-1 (36 mg, 0.1 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 solution, and borate SL-B4 (48 mg, 0.2 mmol) was added thereto. The reaction solution was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (3.5 mg, 0.01 mmol) and 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (11 mg, 0.02 mmol) were added and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography to obtain the target compound SL-ZYE-120. (4.1 mg). 1H NMR (400 MHz, DMSO-d6) δ 9.44 (s, 1H), 8.62 (m, 1H), 7.67 (m, 1H), 7.27 (m, 1H), 6.93 (m, 1H), 6.70 (m, 1H), 4.69 (m, 2H), 4.52 (m, 2H), 3.84 (m, 2H), 3.26 (m 2H), 2.42 (m, 2H), 1.28 (s, 6H). LC-MS: [M+H]+=396.2.
SL-ZYE-119 (10 mg) was dissolved in 2 mL of methanol, and 10% Pd/C (4 mg) was added and the reaction was performed at room temperature for 6 hours. The resultant was filtered and separated by column chromatography to obtain the target compound SL-ZYE-121 (4 mg). 1H NMR (400 MHz, DMSO-d6) δ 9.32 (s, 1H), 8.32 (s, 1H), 7.38 (m, 1H), 6.85 (m, 1H), 6.62 (m, 1H), 4.56-4.47 (m, 4H), 3.68 (m, 2H), 3.20 (m, 3H), 1.70 (m, 4H), 1.09 (m, 6H). LC-MS: [M+H]+=398.2.
Bromide 4-1 (36 mg, 0.1 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 solution, and borate SL-B5 (50 mg, 0.2 mmol) was added thereto. The reaction solution was stirred at room temperature for 10 min under Ar atmosphere protection. 10%6 of allyl palladium (II) chloride dimer (3.5 mg, 0.01 mmol) and 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (11 mg, 0.02 mmol) were added and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography to obtain the target compound SL-ZYE-195 (4.1 mg). 1H NMR (400 MHz, DMSO-d6) δ 9.45 (s, 1H), 8.63 (m, 1H), 7.68 (m, 1H), 7.28 (m, 1H), 6.93 (m, 1H), 6.71 (m, 1H), 4.68 (d, J=4.0 Hz, 2H), 4.54 (J=8.0 Hz, 2H), 3.27 (J=8.0 Hz, 2H), 2.38 (m, 2H), 1.29 (s, 6H), 1.24 (s, 6H). LC-MS: [M+H]+=424.2.
Bromide 4-1 (36 mg, 0.1 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 solution, and borate SL-B6 (49 mg, 0.2 mmol) was added thereto. The reaction solution was stirred at room temperature for 10 min under Ar atmosphere protection. 10% of allyl palladium (II) chloride dimer (3.5 mg, 0.01 mmol) and 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (11 mg, 0.02 mmol) were added and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography to obtain the target compound SL-ZYE-196 (4.1 mg). 1H NMR (400 MHz, DMSO-d6) δ 9.44 (s, 1H), 8.61 (m, 1H), 7.66 (m, 1H), 7.32 (m, 1H), 6.94 (m, 1H), 6.70 (m, 1H), 4.68 (d, J=4.0 Hz, 2H), 4.54 (t, J=8.0 Hz, 2H), 4.34 (m, 2H), 3.70 (m, 1H), 3.29 (J=8.0 Hz, 2H), 2.56-2.21 (m, 2H), 1.25 (d, J=4.0 Hz, 3H). LC-MS: [M+H]+=382.2.
Bromide 4-1 (36 mg, 0.1 mmol) was dissolved in 6 mL of 1,4-dioxane and 2 mL of a 2 M Na2CO3 solution, and borate SL-B7 (49 mg, 0.2 mmol) was added thereto. The reaction solution was stirred at room temperature for 10 min under Ar atmosphere protection. 10%6 of allyl palladium (II) chloride dimer (3.5 mg, 0.01 mmol) and 20% of sodium 2′-dicyclohexylphosphino-2,6-dimethoxy-1,1′-biphenyl-3-sulfonate hydrate (11 mg, 0.02 mmol) were added and the reaction was performed at 90° C. for 40 min under Ar atmosphere protection. After being cooled and concentrated under reduced pressure, the resultant was separated by column chromatography to obtain the target compound SL-ZYE-197 (4.1 mg). 1H NMR (400 MHz, DMSO-d6) δ 9.44 (s, 1H), 8.63 (m, 1H), 7.67 (m, 1H), 7.28 (m, 1H), 6.94 (m, 1H), 6.71 (m, 1H), 4.69 (d, J=4.0 Hz, 2H), 4.53 (t, J=8.0 Hz, 2H), 4.37 (m, 1H), 4.06 (m, 1H), 3.68 (m, 1H), 3.28 (J=8.0 Hz, 2H), 2.60-2.32 (m, 2H), 1.26 (d, J=4.0 Hz, 3H). LC-MS: [M+H]+=382.2.
Compound E-Y3 (46 mg) was dissolved in 5 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Ethylsulfonyl chloride (20 mg) was added at room temperature, and the reaction was continued. When the reaction was completed as indicated by TLC, the resultant was directly separated by column chromatography to obtain the target compound SL-ZYE-144 (7 mg).
1H NMR (400 MHz, CDCl3) δ 9.00 (s, 1H), 7.64 (s, 1H), 7.24 (m, 1H), 6.78 (m, 1H), 6.61 (m, 2H), 4.80 (s, 2H), 4.61 (t, J=8.7 Hz, 2H), 4.06 (m, 2H), 3.61 (t, J=5.6 Hz, 2H), 3.38 (t, J=8.7 Hz, 2H), 3.04 (q, J=7.4 Hz, 2H), 2.72 (m, 2H), 1.40 (t, J=7.4 Hz, 3H). LC-MS: [M+H]+=459.1.
Compound E-Y3 (46 mg) was dissolved in 5 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Cyclopropylsulfonyl chloride (20 mg) was added at room temperature, and the reaction was continued. When the reaction was completed as indicated by TLC, the resultant was directly separated by column chromatography to obtain the target compound SL-ZYE-146 (6 mg).
1H NMR (400 MHz, CDCl3) δ 9.03 (s, 1H), 7.64 (s, 1H), 7.23 (m, 1H), 6.77 (m, 2H), 6.58 (dd, J=8.7, 3.9 Hz, 1H), 4.78 (d, J=5.3 Hz, 2H), 4.60 (t, J=8.7 Hz, 2H), 4.08 (m, 2H), 3.60 (t, J=5.7 Hz, 2H), 3.37 (t, J=8.7 Hz, 2H), 2.74 (m, 2H), 2.42-2.27 (m, 1H), 1.32-1.14 (m, 2H), 1.11-0.88 (m, 2H). LC-MS: [M−H]+=469.2.
Compound E-Y3 (46 mg) was dissolved in 5 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Methyl chloroformate (15 mg) was added at room temperature, and the reaction was continued. When the reaction was completed as indicated by TLC, the resultant was directly separated by column chromatography to obtain the target compound SL-ZYE-147 (5 mg).
1H NMR (400 MHz, DMSO-d6) δ 9.45 (s, 1H), 8.64 (m, 1H), 7.69 (s, 1H), 7.26 (s, 1H), 6.93 (m, 1H), 6.71 (m, 1H), 4.69 (m, 2H), 4.54 (m, 2H), 4.12 (m, 2H), 3.64 (s, 3H), 3.61 (m, 2H), 3.28 (m, 2H), 2.57 (m, 2H). LC-MS: [M+H]+=425.2.
Compound E-Y3 (46 mg) was dissolved in 5 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Ethyl chloroformate (15 mg) was added at room temperature, and the reaction was continued. When the reaction was completed as indicated by TLC, the resultant was directly separated by column chromatography to obtain the target compound SL-ZYE-148 (4 mg).
1H NMR (400 MHz, CDCl3) δ 9.12 (s, 1H), 7.64 (s, 1H), 7.26-7.09 (m, 1H), 7.00 (m, 1H), 6.74 (t, J=9.2 Hz, 1H), 6.57 (dd, J=8.6, 3.9 Hz, 1H), 4.78 (d, J=4.9 Hz, 2H), 4.58 (t, J=8.8 Hz, 2H), 4.23-4.10 (m, 4H), 3.72 (d, J=5.3 Hz, 2H), 3.35 (t, J=9.0 Hz, 2H), 2.63 (m, 2H), 1.29 (t, J=7.0 Hz, 3H). LC-MS: [M+H]+=439.2.
Compound E-Y3 (46 mg) was dissolved in 5 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Propionyl chloride (10 mg) was added at room temperature, and the reaction was continued. When the reaction was completed as indicated by TLC, the resultant was directly separated by column chromatography to obtain the target compound SL-ZYE-161 (4 mg), 1H NMR (400 MHz, CDCl3) δ 9.76 (m, 1H), 8.19 (m, 1H), 7.62 (m, 1H), 7.34-7.13 (m, 1H), 6.82-6.67 (m, 1H), 6.58 (m, 1H), 4.79 (m, 2H), 4.57 (m, 2H), 4.26 (m, 2H), 3.82 (m, 2H), 3.39 (m, 2H), 2.65 (m, 2H), 2.50-2.29 (m, 2H), 1.17 (m, 3H). LC-MS: [M+H]+=423.2.
Compound E-Y3 (46 mg) was dissolved in 5 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Cyclopropionyl chloride (10 mg) was added at room temperature, and the reaction was continued. When the reaction was completed as indicated by TLC, the resultant was directly separated by column chromatography to obtain the target compound SL-ZYE-162 (4 mg). 1H NMR (400 MHz, CDCl3) δ 9.35 (m, 1H), 7.81 (m, 1H), 7.64 (m, 1H), 7.33-7.10 (m, 1H), 6.74 (m, 1H), 6.57 (m, 1H), 4.77 (m, 2H), 4.57 (m, 2H), 4.29 (m, 2H), 3.85 (m, 2H), 3.34 (m, 2H), 2.77-2.60 (m, 2H), 1.84 (m, 1H), 0.85 (m, 4H). LC-MS: [M+H]+=435.2.
Compound E-Y3 (46 mg) was dissolved in 5 mL of dichloromethane (DCM) and 1 mL of trifluoroacetic acid (TFA), and the mixture was stirred at room temperature. After the solvent was concentrated, 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) were added and dissolved by stirring. Isopropylsulfonyl chloride (20 mg) was added at room temperature, and the reaction was continued. When the reaction was completed as indicated by TLC, the resultant was directly separated by column chromatography to obtain the target compound SL-ZYE-145 (7 mg).
1H NMR (400 MHz, DMSO-d6) δ 9.46 (s, 1H), 8.69 (m, 1H), 7.70 (s, 1H), 7.33 (m, 1H), 6.96 (d, J=8.7 Hz, 1H), 6.71 (d, J=3.9 Hz, 1H), 4.69 (m, 2H), 4.55 (d, J=8.7 Hz, 2H), 4.04 (m, 2H), 3.53 (d, J=5.7 Hz, 2H), 3.47-3.37 (m, 1H), 3.32-3.24 (m, 2H), 2.66 (m, 2H), 1.25 (s, 3H), 1.23 (s, 3H). LC-MS: [M+H]+=473.2.
SL-E12 (30 mg) was dissolved in 3 mL of dichloromethane and 0.1 mL of triethylamine (TEA) by stirring. Methyl chloroformate (15 mg) was added at room temperature, and the reaction was continued. When the reaction was completed as indicated by TLC, the resultant was directly separated by column chromatography to obtain the target compound SL-ZYE-183 (5 mg). 1H NMR (400 MHz, DMSO-d6) δ 9.43 (s, 1H), 8.60 (m, 1H), 7.62 (s, 1H), 6.98-6.90 (m, 1H), 6.82 (m, 1H), 6.69 (m, 1H), 4.67 (d, J=5.0 Hz, 2H), 4.53 (t, J=8.7 Hz, 2H), 4.07 (m, 2H), 3.59 (m, 5H), 3.28 (t, J=8.7 Hz, 2H), 2.72 (m, 2H), 1.86 (m, 2H). LC-MS: [M+H]+=439.2.
The TR-FRET method was used to measure the PRC2 enzyme activity. First, the enzyme was mixed with compounds at different concentrations and incubated for 30 min at room temperature. The biotin-labeled histone H3 peptide substrate and the cofactor S-adenosylmethionine (SAM) were added to initiate the enzymatic reaction. After reaction was performed at room temperature for 4 hour, Acceptor and Donor were added and incubated for half an hour. The multifunctional microplate reader EnVision (Perkin Elmer Inc.) was used to detect the fluorescence signal. Data were analyzed using GraphPad Prism 5.0 software, and IC50 values were obtained.
The compounds in Table 1 can be prepared by the methods described in the above examples. EED226 is the positive compound (Nat. Chem. Biol. 2017, 13, 381-388), and N/A represents unanalyzed.
Pfeiffer cells in exponential growth phase were seeded in a 24-well plate with a cell density of 1*10E5 cells/mL. Cells were treated on the same day with the compounds at different concentrations. At day 4 and day 7 of compound treatment, fresh media and compounds were changed. After treated with the compounds for 11 days, cell viability was measured using CellTiter-Glo reagent (Promega Corporation). The data was analyzed using GraphPad Prism 5.0 software and GI50 values were obtained.
The compounds in Table 2 can be prepared by the methods described in the above examples, and EED226 is the positive compound (Nat. Chem. Biol. 2017, 13, 381-388).
In a CO2 cell incubator (37° C., 5% CO2), Human diffuse large B-cell lymphoma (DLBCL) cell line, pfeiffer (obtained from ATCC, CRL-2632) were incubated with RPMI 1640 medium (Gibco, purchased from Life Technologies, 22400-089) containing 10% fetal bovine serum (Gibco, purchased from Life Technologies, 10099-141) and 1% antibiotics (penicillin and streptomycin, purchased from Life Technologies, 10378016). In the Experiment on growth inhibition of cells in a long term, pfeiffer cells in exponential growth phase were seeded in 24-well plates (purchased from Corning Corporation, 3524) with a volume of 1 mL/well and a cell density of 2*10E5 cells/mL. After seeding the cells, they were placed in a CO2 incubator and allowed to stand for 1 hour. In a 24-well plate containing cells, 2 μL of three-fold gradient of the compounds at 9 different concentrations or DMSO were added to each well, such that the final concentration of the compounds was 0.003-20 μM or 0.3-2000 nM, and the final concentration of DMSO was 0.2%. At day 4, day 7, and day 11 of compound treatment, the fresh medium and compounds were changed, and the cell density of the DMSO control well was diluted to 2*10E5 cells/mL. The cell dilution ratio for other compound wells was the same as that of the DMSO control well. Cell viability was determined by using CellTiter-Glo reagent (purchased from Promega Inc., G7572): cells treated with the compounds for 14 days were transferred to a white 384-well plate (OptiPlate-384, purchased from PerkinElmer, 6007299) at 40 μL/well, an equal volume of CellTiter-Glo reagent was further added. After incubating at room temperature for 10 min, the cold luminescence signal was detected with a multifunctional microplate reader EnVision (PerkinElmer) at a wavelength of 400 to 700 nm. Data were analyzed using GraphPad Prism 5.0 software, and IC50 values were obtained.
The compounds in Table 3 can be prepared by the methods described in the above examples, and EED226 is the positive compound (Nat. Chem. Biol. 2017, 13, 381-388).
In a CO2 cell incubator (37° C., 5% CO2), Human diffuse large B-cell lymphoma (DLBCL) cell line, Karpas-422, SU-DHL-4 (obtained from ATCC, CRL-2957) were incubated with RPMI 1640 medium (Gibco, purchased from Life Technologies, 22400-089) containing 10% fetal bovine serum (Gibco, purchased from Life Technologies, 10099-141) and 1% antibiotics (penicillin and streptomycin, purchased from Life Technologies, 10378016). In the Experiment on growth inhibition of cells in a long term, Karpas-422, SU-DHL-4 cells in exponential growth phase were seeded in 24-well plates (purchased from Corning Corporation, 3524) with a cell density of 1*10E5/mL and a cell medium volume of 1 mL. After incubating in a 24-well plate for 1 hour, 2 μL of the compounds or DMSO were added to each well. For each compound, there were 9 different concentrations, such that the final concentration in the cell medium was 0.003-20 μM or 0.3-2000 nM, and the final concentration of DMSO was 0.2%. At day 4, day 7 of compound treatment, the fresh cell medium and compounds were changed, and the cell density of the DMSO control well was diluted to 1*10E5/mL. The cell dilution ratio for compound wells was the same as that of the DMSO control well. Cell viability was determined by using CellTiter-Glo reagent (purchased from Promega Inc., G7572): cells treated with the compounds for 11 days were transferred to a white 384-well plate (OptiPlate-384, purchased from PerkinElmer, 6007299) at 40 μL/well, an equal volume of CellTiter-Glo reagent was further added. After incubating at room temperature for 10 min, the cold luminescence signal was detected with a multifunctional microplate reader EnVision (purchased from PerkinElmer) at a wavelength of 400 to 700 nm. Data were analyzed using GraphPad Prism 5.0 software, and IC50 values were obtained.
The compounds in Table 4 can be prepared by the methods described in the above examples, and EED226 is the positive compound (Nat. Chem. Biol. 2017, 13, 381-388).
From the data in the above Tables 1 to 4, it can be seen that the IC50 value of some compounds of the present invention on PRC2 enzyme can be up to a nM level, which is significantly higher than that of the positive control compound EED226; similarly, in the experiments on growth inhibition of Pfeiffer, Karpas-422, and SU-DHL-4 cells in a long term, the IC50 values of several compounds of the present invention also is up to a nM level in single digit, which are significantly higher than the positive control EED226 compound.
1. Healthy male SD rats were used as test animals, EED226, E-Y1, E-Y13, E-Y47, SL-ZYE-07 (3 mg/kg) were administered intragastrically, and the drug concentrations in rat plasma were determined by LC/MS/MS method at different time points after drug administration. The pharmacokinetic behavior of the compounds of the present invention in rats was studied, and the pharmacokinetic properties were evaluated.
2. The test animals were healthy adult male SD rats, with 3 rats in each group, purchased from Shanghai Xipuer-Bikai Laboratory Animal Limited Company.
3. Drug preparation: Compounds EED226, E-Y1, E-Y13, E-Y47 were dissolved in DMSO/0.5% HPMC (5/95, v/v) for preparation. The compound SL-ZYE-07 was dissolved in 0.5% HPMC (containing 0.5% Tween 80), vortexed, and sonicated to disperse the solid matter uniformly to obtain a pale white suspension.
4, Operation: EED226, E-Y1, E-Y13, E-Y47, SL-ZYE-07 were administered to rats by intragastric administration (3 mg/kg), 45 μL of blood was taken through the femoral vein at 0.25, 0.5, 1, 2, 4, 8, 24 hour after administration, and centrifuged in a heparinized centrifuge tube for 5 min, and the plasma samples were separated for analysis. The contents of test compounds in rat plasma after intragastric administration of different compounds were determined by Liquid chromatography-mass spectrometry (LC-MS/MS).
The pharmacokinetic parameters of the compounds of the present invention are as follows:
Conclusion: The compounds of the present invention show good pharmacokinetic absorption and have prominent advantages in pharmacokinetic.
Compound Information
Stock Solutions of Compounds E-Y1, E-Y13, E-Y15, E-Y40, E-Y43, E-Y47, E-Y50, E-Y54, E-Y54-H, SL-E23 (10 mM in DMSO)
1. JANUS and temperature control device was turned on. After being initialized, the pipes were flushed until there was no air bubble in the pipes.
2. Preparation of buffers used for the experiment
Preparation of Tris pH 7.4 buffer (0.1 M): 12.12 g of Tris was dissolved in 1000 mL of H2O, the resulting solution was adjusted to pH 7.4 with 2N HCl, aliquoted at 50 mL per tube, and stored at −20° C.
Preparation of H2O/0.1% BSA buffer: 200 μL of 25% BSA was added to 50 mL of H2O.
Preparation of VIVID stock solution: 1 mg of VIVID was dissolved in 1 mL of acetonitrile, aliquoted at 50 μL per tube, and stored at −20° C.
Preparation of MgCl2 solution (100 mM): 1.016 g of MgCl2 was dissolved in 50 mL Tris pH 7.4 buffer, and aliquoted at 1 mL per tube, and stored at −20° C.
Preparation of NADPH solution (10 mM): 355 mg of NADPH was dissolved in 42.6 mL of Tris pH 7.4 buffer, aliquoted at 1.8 mL per tube, and stored at −20° C.
Preparation of Blank control: 7.937 mL of Tris, 163 μL of RLM, 450 μL of MgCl2 solution, 450 μL of NADPH solution and 9 mL of methanol were mixed uniformly, aliquoted at 1 mL per tube, and stored at −20° C.
3. Preparation of Compound Working Solution
Dilution step 1:10 μL of compound stock solution was added to 90 μL of DMSO=1 mM stock solution.
Dilution step 2: 2 μL of 1 mM stock solution was added to 1 mL of H2O/0.1%6 BSA buffer=0.2 μM working solution.
Dilution step 3: 245 μL of the working solution was added to a 96-well compound plate and added with 5 μL of VIVID stock solution.
The 96-well compound plate was shaken on a shaker for 5 min.
4. The STM in the JANUS in the computer was open to run the EXCEL file, and the JANUS program was operated and run by following the instructions in the EXCEL file.
5. After the JANUS program was finished, acetonitrile/water 50:50 (V/V) was added to column 20 of the 384-well black plate, and blank control was added to column 21 of the 384-well black plate.
6. the plate (420 nm-465 nm) was read on a microplate reader to determine the VIVID fluorescence intensity.
7. After the plate was sealed, shaken and centrifuged, the sample plate was subjected to the LC/MS instrument for sample analysis.
Data Analysis:
A profile of the logarithmic value of the remaining rate of the drug in the incubation system versus the incubation time was plotted, and the slope k was obtained by linear regression. The intrinsic clearance rate in vivo (Clint, mL/min/g) value, the clearance rate in vivo (Cl, mL)/min), liver clearance (Clhep, mL/min) and metabolic bioavailability (% MF) were predicted according to the following equations:
Results of Liver Microsomal Stability of the Compounds:
Conclusion: The compounds of the present invention have good liver microsomal stability in human, rat, and mouse, and thus having prominent advantages.
| Number | Date | Country | Kind |
|---|---|---|---|
| 201710898099.5 | Sep 2017 | CN | national |
| 201711408714.6 | Dec 2017 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2018/102833 | 8/29/2018 | WO | 00 |
| Publishing Document | Publishing Date | Country | Kind |
|---|---|---|---|
| WO2019/062435 | 4/4/2019 | WO | A |
| Number | Name | Date | Kind |
|---|---|---|---|
| 9580437 | Chan | Feb 2017 | B2 |
| Number | Date | Country |
|---|---|---|
| 2016103155 | Jun 2016 | WO |
| Entry |
|---|
| Huang et al., “Discovery of First-in-Class, Potent, and Orally Bioavailable Embryonic Ectoderm Development (EED) Inhibitor with Robust Anticancer Efficacy,” J. Med. Chem., vol. 60, Jan. 16, 2017, pp. 2215-2226. |
| Yang et al., “Allosteric Inactivation of Polycomb Repressive Complex 2 (PRC2) by Inhibiting Its Adapter Protein: Embryotic Ectodomain Development (EED),” J. Med. Chem., vol. 60, Mar. 3, 2017, pp. 2212-2214. |
| Qi et al., “An Allosteric PRC2 Inhibitor Targeting the H3K27me3 Binding Pocket of EED,” Nature Chemical Biology, vol. 13, Jan. 30, 2017, pp. 381-388. |
| International Search Report and Written Opinion for International Application No. PCT/CN2018/102833 dated Nov. 30, 2018. |
| International Preliminary Report on Patentability for International Application No. PCT/CN2018/102833 dated Mar. 31, 2020. |
| Number | Date | Country | |
|---|---|---|---|
| 20200261459 A1 | Aug 2020 | US |
