Cyclin modulator

Abstract

The present invention provides a cyclin modulator. Specifically, the present invention provides a compound as shown in Formula (I) or a pharmaceutically acceptable salt thereof.

Description

Cell cycle regulators Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and in particular relates to a cell cycle protein regulator. Background Art

[0002] Unlike most traditional drugs that directly inhibit the action of molecular targets, molecular glue degraders kill cancer cells by destroying target proteins through the ubiquitin-proteasome system. For example, the multiple myeloma drug lenalidomide is a molecular glue degrader that recruits E3 ubiquitin ligase to mark the target protein in the cell and then degrade it. Unlike traditional small molecule inhibitors, molecular glues drive target ubiquitination and induce degradation in a catalytic manner, which is a new strategy that can inactivate targets that are difficult to treat with traditional pharmacological methods. Molecular glues also cleverly circumvent the limitations of traditional inhibitors, turning some targets from "non-drugable" to "drugable."

[0003] The cell cycle is a fundamental process in cellular life, controlling the transition of cells from a quiescent phase to a proliferative phase. Cyclin-dependent kinases (CDKs) and cyclins are core molecules in the entire cell cycle regulatory mechanism. In normal cells, the activity of cyclins is strictly controlled by their cell cycle-specific transcription and protein degradation, as well as the influence of certain CDK inhibitory proteins. In addition to promoting cell division, cyclins are also responsible for regulating various cellular functions. This mechanism is jointly operated by cyclins and their catalytic partners, cyclin-dependent kinases (CDKs). However, these influencing factors often become uncontrolled in human cancers, leading to abnormal activation of cyclins. Abnormal activity of the cell cycle mechanism is present in virtually all types of tumors and is a driving force in tumorigenesis. Targeting specific cyclins may become an effective anti-cancer strategy.

[0004] To date, only a few CDK inhibitors have been marketed, all of which are CDK4 / 6 inhibitors. Inhibitors that selectively target other CDK families struggle to achieve the required specificity and are almost all multi-target inhibitors, exhibiting unacceptable toxicity in clinical practice, thus hindering their clinical application. Furthermore, while kinase inhibitors can lead to the removal of kinase subunits, they simultaneously maintain the integrity of cell cycle proteins, potentially triggering compensatory mechanisms. Kinase degradation can lead to effects that outlast the inhibitory effect.

[0005] While molecular glue degraders are highly desirable, clinically effective, and highly sought-after, few have been discovered to date, and most have been discovered by chance. CR8, a newly discovered cyclin K (Cyclin K) degrader, is primarily a multikinase inhibitor that inhibits the activity of multiple cyclins in the CDK family, resulting in toxicity that limits its clinical application. In addition to its poor selectivity, its activity in degrading Cyclin K is also modest, limiting its use in catalytic amounts.

[0006] In summary, there is an urgent need in the art to develop a class of cell cycle protein regulators such as cell cycle protein degraders with higher activity and / or lower toxicity. Summary of the Invention

[0007] The purpose of the present invention is to provide a cell cycle protein degrader with higher activity, or a novel cell cycle protein regulator.

[0008] In the first aspect of the present invention, a compound or a pharmaceutically acceptable salt thereof is provided, wherein the compound is represented by formula (I):

[0009] in,

[0010] R 1 Each independently is H or C 1-4 alkyl;

[0011] Subscript n1 is 1, 2, or 3;

[0012] Cyclic Ar 1 Select from the following group: C 6-10 Aromatic ring, 5- to 10-membered heteroaromatic ring, 5- to 10-membered bridged ring;

[0013] Cyclic Cr 1 Selected from the following group: H, C 3-10 Carbocyclic group, 3 to 10 membered heterocyclic group, C 6-10 aryl, and 5- to 10-membered heteroaryl;

[0014] R a and R b Each independently selected from the group consisting of: H, R e or R; or R a and R b With cyclic Ar 1 and cyclic Cr 1 Together in,

[0015] X 7 Each independently selected from the group consisting of: -O-, -S-, -N(R c )-、-C(R c )2-、-C(Rc )2-C(R c )2-;

[0016] Subscripts n5 and n6 are each independently 0, 1, 2 or 3;

[0017] R e Each independently selected from the group consisting of: hydroxyl, C 1-6 Alkyl, -OC 1-6 Alkyl, -OC 1-6 Alkylene-R f ;

[0018] Among them, R f Selected from the group consisting of: -CN, -OH, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2;

[0019] Subscripts n3 and n4 are each independently 0, 1, 2, 3 or 4;

[0020] R 2 Selected from the group consisting of H, CN, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-8 cycloalkyl, optionally substituted 3- to 8-membered heterocyclyl;

[0021] X 1 and X 2 Each independently is N or C(R c );

[0022] X 3 、X 4 and X 5 are each independently N or C;

[0023] X 6 N, N(R c )、C(R c ), O or S;

[0024] M 1 Select from the following groups: None, X 8 、(M 4 ) s ;in,

[0025] X 8 N(R c ) or C(R c )2;

[0026] M 4 Each independently selected from the group consisting of O, S, C(O)O, C(O), N(Rc ) and C 1-4 alkylene;

[0027] s is 1, 2, or 3;

[0028] M 2 is none or a ring as shown in formula A;

[0029] In formula A,

[0030] X 9 Representative and M 1 Connected position, and X 9 N or C(R m );

[0031] X 10 Representative and M 3 Connected position, and X 10 Selected from the group consisting of O, S, N, or C(R m );

[0032] Each X 11 are independently -C(R m )2-or-N(R m )-;

[0033] Subscripts m1 and m2 are each independently 0, 1, 2 or 3, and m1+m2≥2;

[0034] R m Each independently is R c or R 5 ; or, two R m Together form a single bond, optionally substituted C 1-4 alkylene or optionally substituted 1- to 4-membered heteroalkylene;

[0035] R 5 Each independently selected from the group consisting of: hydroxy, optionally substituted C 1-6 Hydroxyalkyl, optionally substituted -C(O)-NH2, optionally substituted -C(O)-NH(C 1-6- alkyl), optionally substituted -C(O)-N(C 1-6- alkyl)2, optionally substituted C 1-6 Alkyl, optionally substituted -C(O)-C 1-6- Alkyl and optionally substituted C 1-6 haloalkyl; or, two R on the same carbon atom 5 Together they form an oxo group (=O) or a thio group (=S);

[0036] M 3 Selected from the group consisting of: H, None, R 3 、-NH-R4 ;in,

[0037] R 3 Selected from the group consisting of hydroxy, optionally substituted C 1-6- Hydroxyalkyl, optionally substituted -C(O)-C 1-6- alkyl, optionally substituted-C(O)-NH2, optionally substituted-C(O)-NH(C 1-6- alkyl), optionally substituted -C(O)-N(C 1-6- alkyl)2, optionally substituted C 1-6- alkyl halide;

[0038] R 4 Selected from the group consisting of: H, optionally substituted C 1-6 Alkyl, optionally substituted C 1-6- Hydroxyalkyl, optionally substituted C 1-6- alkyl halide;

[0039] R c Each independently is H or C 1-4 alkyl;

[0040] Unless otherwise defined, the optional substitution refers to unsubstituted or one or more (such as 1, 2, 3 or 4) hydrogen in the group is replaced by a substituent R, and R is selected from the following groups: D, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -CN, -OR', -NO2, -NR'R", -SR', -OC(O)R', -C(O)R', -CO2R', -CONR', -OC(O)NR'R", -NR"C(O)R', -NR"-C(O)NR'R", -NR"C(O)2R', -S(O)R', -S(O)2R', -S(O)2NR'R", -NR"S(O)2R', C(O)2R', optionally substituted with one or more R' 3-10 Cycloalkyl, 4 to 10 membered heterocycloalkyl optionally substituted by one or more R'", C 6-10 aryl, 5- to 10-membered heteroaryl optionally substituted by one or more R'", -C 1-4 Alkylene-C 3-10 Cycloalkyl, optionally substituted with one or more R'"-C 1-4 Alkylene-4 to 10 membered heterocycloalkyl, optionally substituted with one or more R'"-C 1-4 Alkylene-C 6-10 Aryl, optionally substituted with one or more R'" -C1-4 Alkylene-5 to 10 membered heteroaryl;

[0041] Each R' is independently H, D, or a group selected from the group consisting of: C 1-6 Alkyl, C 3-10 Cycloalkyl, 4- to 10-membered heterocycloalkyl, C 6-10 Aryl, 5- to 10-membered heteroaryl, -C 1-4 Alkylene-C 3-10 Cycloalkyl, -C 1-4 Alkylene-4 to 10 membered heterocycloalkyl, -C 1-4 Alkylene-C 6-10 Aryl-C 1-4 Alkylene-5 to 10 membered heteroaryl;

[0042] Each R" is selected from the group consisting of H, D, C 1-4 Alkyl, C 1-4 Haloalkyl, and C 3-4 Cycloalkyl;

[0043] Each R"' is independently selected from the group consisting of D, halogen, hydroxy, nitro, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl.

[0044] In another preferred embodiment, the compound or a pharmaceutically acceptable salt thereof is used as a molecular glue degrading agent.

[0045] In another preferred embodiment, the compound is not CR8 as shown below:

[0046] In another preferred embodiment, the compound does not include the compounds in the following table:

[0047] In another preferred embodiment, R 1 is H. In another preferred embodiment, n1=1. In another preferred embodiment, R 1 is H, and n1=1. In another preferred embodiment, ring Ar 1 Select from the following group: C 6-10 aromatic ring, 5- to 10-membered heteroaromatic ring.

[0048] In another preferred embodiment, ring Ar 1 Selected from the group consisting of a benzene ring and a 5- to 10-membered heteroaromatic ring.

[0049] In another preferred embodiment, ring Ar 1 Select from the following groups:

[0050] Among them, * represents the ring Cr 1 Connection location; X a 、X b 、X c and X d are independently CH and N; X g Selected from the group consisting of NH, O, S; X h 、X i and X j Each is independently -CH2- or -CH2-CH2-.

[0051] In another preferred embodiment, for

[0052] In another preferred embodiment, ring Ar 1 middle, for

[0053] In another preferred embodiment, ring Ar 1 Select from the following groups:

[0054] Among them, * represents the ring Cr 1 Connection location; X a 、X b 、X c and X d are independently CH and N; X g Selected from the group consisting of NH, O, S.

[0055] In another preferred embodiment, ring Ar 1 for

[0056] Among them, * represents the ring Cr 1 The location of the connection.

[0057] In another preferred embodiment, for Where * represents the ring Cr 1 Connection location

[0058] In another preferred embodiment, the ring Cr 1 In the embodiment, the carbocyclic ring is a saturated or unsaturated carbocyclic ring containing 1 or 2 double bonds.

[0059] In another preferred embodiment, the ring Cr 1 In the C 3-10 Carbon ring is C 4-10 Carbocyclic ring; preferably, C 4-6 Carbon ring.

[0060] In another preferred embodiment, the ring Cr1 In the embodiment, the 3- to 10-membered heterocyclic group is a saturated 3- to 10-membered heterocyclic group.

[0061] In another preferred embodiment, the ring Cr 1 In the embodiment, the 3- to 10-membered heterocyclic group is a 4- to 10-membered heterocyclic group; preferably, it is a 4- to 6-membered heterocyclic group.

[0062] In another preferred embodiment, the ring Cr 1 Select from the following groups:

[0063] Among them, X d and X e are each independently N or CH; X f NH, S, O; X g is N or CH.

[0064] In another preferred embodiment, for

[0065] In another preferred embodiment, the ring Cr 1 Ring Ar 2 ; and cyclic Ar 2 Select from the following group: C 6-10 aryl, and 5- to 10-membered heteroaryl.

[0066] In another preferred embodiment, ring Ar 2 Select from the following groups:

[0067] Among them, X d and X e are each independently -N- or -CH-; X f is -NH-, -S-, -O-; X g is N or CH.

[0068] In another preferred embodiment, ring Ar 2 for In another preferred embodiment, ring Ar 2 It is phenyl.

[0069] In another preferred embodiment, when M 2 When it is nothing, the ring Ar 2 It is not a nitrogen-containing heteroaryl group (wherein the nitrogen-containing heteroaryl group is a heteroaryl group having 1 or 2 nitrogen heteroatoms in the ring and no other heteroatoms, such as pyridyl, pyrazolyl, imidazolyl and pyrazinyl).

[0070] In another preferred embodiment, when M2 is absent, the ring Ar 2 Not for

[0071] In another preferred embodiment, n3 is 0 (ie, ring Ar 1 is unsubstituted); or, n3 is 1, 2, 3 or 4 (i.e., ring Ar 1 1, 2, 3 or 4 R a substituted), and R a Each independently selected from the group consisting of: D, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl.

[0072] In another preferred embodiment, n3 is 0. In another preferred embodiment, n3 is 1, and R a R e .

[0073] In another preferred embodiment, n4 is 0 (ie, ring Cr 1 is unsubstituted); or, n4 is 1, 2, 3 or 4 (i.e., ring Cr 1 1, 2, 3 or 4 R b substituted), and R b Each independently selected from the group consisting of: D, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl.

[0074] In another preferred embodiment, n4 is 0. In another preferred embodiment, n4 is 1, and R b R e .

[0075] In another preferred embodiment, n3 is 0 (ie, ring Ar 1 is unsubstituted); or, n3 is 1, 2, 3 or 4 (i.e., ring Ar 1 1, 2, 3 or 4 R a substituted), and R a Each independently selected from the group consisting of: D, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl.

[0076] In another preferred embodiment, when M2 is zero, Not for

[0077] In another preferred embodiment, for

[0078] In another preferred embodiment, the compound is as shown in formula (II);

[0079] Among them, X 1 、X 2 、X 3 、X 4 、X 5、X 6 、M 1 、M 2 、M 3 、R 2 As defined above.

[0080] In another preferred embodiment, M 1 None or X 8 (Preferably none or N(R c )).

[0081] In another preferred embodiment, M 1 is None or NH.

[0082] In another preferred embodiment, M 2 is a ring as shown in formula A.

[0083] In another preferred embodiment, X 9 N or C(R c ); preferably X 9 is N or CH.

[0084] In another preferred embodiment, X 10 N or C(R m ); preferably, X 10 N or C(R c ); better X 10 is N or CH.

[0085] In another preferred embodiment, 2≤m1+m2≤4 (ie, the ring shown in formula A is a 4- to 6-membered ring).

[0086] In another preferred embodiment, m1 and m2 are each independently 1 or 2; preferably, m1=2, m2=2.

[0087] In another preferred embodiment, in formula A, when two R m Together form a single bond, optionally substituted C 1-4 When the two R m Located on the same carbon atom, the two R m Together they form an optionally substituted C 2-4 Alkylene or optionally substituted 2 to 4 membered heteroalkylene (ie, the ring shown in formula A is a spiro ring); or, if both R m Located on two adjacent ring atoms, the two R m Together they form an optionally substituted C 1-4 Alkylene or optionally substituted 1 to 4 membered heteroalkylene (ie, the ring shown in formula A is a cyclic ring); or, if the two R m Located on two ring atoms separated by at least one ring atom, then the two R m Together form a single bond, optionally substituted C1-3 Alkylene or optionally substituted 1 to 3 membered heteroalkylene (ie, the ring shown in formula A is a cyclic ring); preferably, the two R m Together they form C 1-4 Alkylene.

[0088] In another preferred embodiment, each X 11 All are -C(R m )2-.

[0089] In another preferred embodiment, R m Each independently is R c or R 5 .

[0090] In another preferred embodiment, X 11 There are at most two R m R 5 , the rest R m All R c (Preferably, the remaining R m All are H).

[0091] In another preferred embodiment, R 5 Each independently selected from the group consisting of: optionally substituted C 1-6 Hydroxyalkyl, optionally substituted -C(O)-NH2, and optionally substituted C 1-6 haloalkyl; or, two R on the same carbon atom 5 Together form an oxo group (=O); preferably, R 5 Each independently selected from the group consisting of: optionally substituted C 1-6 Hydroxyalkyl, and optionally substituted -C(O)-NH2, or two R on the same carbon atom 5 Together they form an oxo group (=O).

[0092] In another preferred embodiment, X 11 0, 1 or 2 of them are each independently -C(R c )(R 5 )-(preferably -CH(R 5 )-), -C(O)- or -C(S)-, the rest X 11 All are -C(R c )2-(preferably, -CH2-).

[0093] In another preferred embodiment, R c For H.

[0094] In another preferred embodiment, R 5 is optionally substituted C 1-6 hydroxyalkyl or optionally substituted -C(O)-NH2.

[0095] In another preferred embodiment, R 5 is optionally substituted C 1-6 Hydroxyalkyl; preferably, -CH2OH.

[0096] In another preferred embodiment, M 3 H or R 3 .

[0097] In another preferred embodiment, R 3 Selected from the group consisting of hydroxy, optionally substituted C 1-6- Hydroxyalkyl.

[0098] In another preferred embodiment, R 3 It is a hydroxyl group.

[0099] In another preferred embodiment, when M 1 is none and M 3 When H, X 11 1 or 2 (preferably 1) R m R 5 , the rest R m All R c (Preferably, the remaining R m All are H).

[0100] In another preferred embodiment, when M 1 is none and M 3 When H, X 11 One of them is -C(R c )(R 5 )-(Preferably, -CH(R 5 )-), -C(O)- or -C(S)-, the rest X 11 and X 12 All are -C(R c )2-(preferably, -CH2-).

[0101] In another preferred embodiment, when M 1 NH and M 3 R 3 When each X 11 All are -C(R c )2-(preferably, -CH2-).

[0102] In another preferred embodiment, for (Preferably, More preferably, ); where X 9 N; X 10 N or C(R m )(Preferably, N or CH; more preferably, N).

[0103] In another preferred embodiment, for Preferably, Wherein Y is O or S.

[0104] In another preferred embodiment, for

[0105] In another preferred embodiment, for

[0106] In another preferred embodiment, for Preferably,

[0107] In another preferred embodiment, for (Preferably, More preferably, ); where X 9 is N or CH (preferably, CH); X 10 is N or CH (preferably, CH).

[0108] In another preferred embodiment, for

[0109] In another preferred embodiment, Select from the following groups:

[0110] In another preferred embodiment, for

[0111] In another preferred embodiment, the compound is as shown in formula II-1 or II-2

[0112] Among them, R 2 、R 3 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 9 、X 10 and X 11 As defined above.

[0113] In another preferred embodiment, in Formula II-1, X 9N, X 10 N, X 11 One of them is -CH(R 5 )-), -C(O)- or -C(S)-, the rest X 11 It is -CH2-.

[0114] In another preferred embodiment, in Formula II-1, R 5 is optionally substituted C 1-6 hydroxyalkyl or optionally substituted -C(O)-NH2.

[0115] In another preferred embodiment, in Formula II-1, R 5 is optionally substituted C 1-6 Hydroxyalkyl; preferably, -CH2OH.

[0116] In another preferred embodiment, in Formula II-2, X 9 CH, X 10 CH, X 11 All -CH2-.

[0117] In another preferred embodiment, R 3 Selected from the group consisting of hydroxy, optionally substituted C 1-6- Hydroxyalkyl; preferably, R 3 It is a hydroxyl group.

[0118] In another preferred embodiment, X 1 N or C(R c ); preferably, N or CH.

[0119] In another preferred embodiment, X 2 N or C(R c ); preferably, N or CH.

[0120] In another preferred embodiment, X 3 N or C.

[0121] In another preferred embodiment, X 4 N or C.

[0122] In another preferred embodiment, X 5 N or C.

[0123] In another preferred embodiment, X 6 N, N(R c ) or C(R c ); preferably, N, NH or CH.

[0124] In another preferred embodiment, X 1 N, X 2 N or C(R c )(Preferably N or CH).

[0125] In another preferred embodiment, X 1 N or C(R c )(Preferably N or CH), X 2 is N.

[0126] In another preferred embodiment, X 1 N, X 2 is N.

[0127] In another preferred embodiment, R c Both are H.

[0128] In another preferred embodiment,

[0129] Select from the following groups:

[0130] In another preferred embodiment, Select from the following groups:

[0131] In another preferred embodiment, R 2 is optionally substituted C 1-6 In another preferred embodiment, R 2 C 1-6 In another preferred embodiment, R 2 is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl.

[0132] In another preferred embodiment, R 1 、R 2 、R 3 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、M 1 、M 2 、M 3 、M 4 , subscript n1, subscript n3, subscript n4, subscript n5, subscript n6, subscript m1, subscript m2, subscript m3, subscript s, Ar 1 、Ar 2 Cr 1 、R a 、R b 、Rc 、R e 、R f 、R m 、R 5 , R, R', R" and R'" are each independently a corresponding group in the Example compounds or the specific compounds in Table A, Table B1 and Table B2.

[0133] In another preferred embodiment, the compound is a compound selected from Table A.

[0134] In another preferred embodiment, the compound is a compound selected from Table B1 and Table B2.

[0135] In a second aspect of the present invention, a pharmaceutical composition is provided, comprising:

[0136] (i) the compound according to the first aspect or a pharmaceutically acceptable salt thereof; and

[0137] (ii) a pharmaceutically acceptable carrier.

[0138] In the third aspect of the present invention, there is provided a use of the compound of formula (I) or a pharmaceutically acceptable salt thereof as described in the first aspect in the preparation of a medicament for treating cancer.

[0139] In a fourth aspect of the present invention, a method for treating cancer is provided, comprising the steps of administering a safe and effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof as described in the first aspect to a subject in need.

[0140] In a fifth aspect of the present invention, there is provided a use of the compound of formula (I) or a pharmaceutically acceptable salt thereof as described in the first aspect in the preparation of a cell cycle protein degrader.

[0141] In another preferred embodiment, the cell cycle protein is cyclin K.

[0142] In a sixth aspect of the present invention, a method for degrading cell cycle proteins is provided, comprising the steps of treating a subject with the compound represented by formula (I) as described in the first aspect, thereby degrading the cell cycle proteins.

[0143] In another preferred embodiment, the cell cycle protein is cyclin K.

[0144] In another preferred embodiment, the object is a cell.

[0145] In another preferred embodiment, the object is HepG2 cells.

[0146] In another preferred embodiment, the method is non-therapeutic in vitro.

[0147] In the seventh aspect of the present invention, a conjugate or prodrug is provided, wherein the conjugate or prodrug is a compound comprising a molecular glue degrading agent portion or a pharmaceutically acceptable salt thereof, wherein the molecular glue degrading agent portion is derived from the compound as described in the first aspect.

[0148] In another preferred embodiment,

[0149] (i) The conjugate is a compound represented by Formula III or a pharmaceutically acceptable salt thereof; D -M CL -M CP (III)

[0150] Among them, M D It is the molecular glue degradation agent part; M CL No or connector part; M CP is a portion derived from a polypeptide element or a targeting ligand;

[0151] (ii) the prodrug is a compound of formula IV or a pharmaceutically acceptable salt thereof; D -M PL -M LG (IV)

[0152] Among them, M D It is the molecular glue degradation agent part; M PL Is none or a connecting group; M LG For the leaving part.

[0153] In another preferred embodiment, the conjugate or prodrug is a compound selected from Table C or a pharmaceutically acceptable salt thereof.

[0154] In an eighth aspect of the present invention, a pharmaceutical composition is provided, comprising:

[0155] (i) the conjugate according to the seventh aspect or a pharmaceutically acceptable salt thereof; and

[0156] (ii) a pharmaceutically acceptable carrier.

[0157] In the ninth aspect of the present invention, there is provided a use of the conjugate according to the seventh aspect or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cancer.

[0158] In the tenth aspect of the present invention, a method for treating cancer is provided, comprising the step of administering a safe and effective amount of the conjugate or a pharmaceutically acceptable salt thereof as described in the sixth aspect to a subject in need.

[0159] In the eleventh aspect of the present invention, there is provided a use of the conjugate according to the seventh aspect or a pharmaceutically acceptable salt thereof in the preparation of a cyclin degrader.

[0160] In another preferred embodiment, the cell cycle protein is cyclin K.

[0161] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0162] none DETAILED DESCRIPTION

[0163] After extensive and in-depth research, the inventors unexpectedly discovered a class of compounds with novel structures (such as those represented by Formula (I), Formula (II), Formula (II-1), and Formula (II-2) herein) that, by modifying functional groups elsewhere in the parent nucleus, also exhibit excellent effects in inducing cyclin K degradation. Based on this discovery, the inventors completed the present invention.

[0164] the term

[0165] Unless otherwise indicated, the bonds represented by dashed lines in each structural formula represent the points of attachment to other moieties.

[0166] As used herein, unless otherwise defined, the term "alkyl" by itself or as part of another substituent refers to a straight or branched chain hydrocarbon radical (i.e., C 1-6 Preferably, the alkyl group has 1 to 4 carbon atoms, i.e., C 1-4 Alkyl. Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc. The term "alkenyl" refers to an unsaturated alkyl group having one or more double bonds. Preferably, alkenyl groups have 2 to 4 carbon atoms, i.e., C 2-4 Similarly, the term "alkynyl" refers to an unsaturated alkyl group having one or more triple bonds. Preferably, the alkynyl group has 2 to 4 carbon atoms, i.e., C 2-4 Examples of such unsaturated alkyl groups include, but are not limited to, ethenyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologs and isomers.

[0167] Unless otherwise indicated, the term "heteroalkyl" by itself or in combination with other terms refers to a stable straight or branched chain group in which one or more (e.g., 1 or 2) carbon atoms of an alkyl group as defined above are replaced by a heteroatom, such as one selected from O, N, Si, and S, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen heteroatom is optionally quaternized. The heteroatoms O, N, and S may be located at any interior position of the heteroalkyl group. The heteroatom Si may be located at any position of the heteroalkyl group, including the position at which the alkyl group is attached to the remainder of the molecule.

[0168] The term "alkylene" by itself or as part of another substituent refers to a divalent group derived from an alkane, for example -CH2-. Preferably, in this application, a alkylene group having 1, 2, 3 or 4 carbon atoms (i.e., C 1-4 alkylene).

[0169] The term "heteroalkylene" by itself or as part of another substituent refers to a divalent radical derived from heteroalkyl.

[0170] As used herein, the term "carbocyclyl" or "carbocycle" refers to a carbocyclic ring having the specified number of ring atoms (e.g., C 3-10 Carbocyclic ring, C 4-10 Carbocyclic ring, C 4-6 The term "heterocyclyl" or "heterocycle" refers to a hydrocarbon ring (radical) that is fully saturated or has one or two double bonds between the ring vertices. The term also includes bicyclic and polycyclic hydrocarbon rings, such as bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. The term "heterocyclyl" or "heterocycle" refers to a carbocyclic ring (radical) containing 1 to 5 heteroatoms selected from N, O and S, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atom is optionally quaternized. The heterocyclic ring (radical) can be a monocyclic, bicyclic or polycyclic ring system, preferably a monocyclic ring. Non-limiting examples of heterocyclic rings include pyrrolidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, quinuclidine, etc. The heterocyclic ring can be attached to the rest of the molecule through a ring carbon or a heteroatom.

[0171] The term "cycloalkyl" refers to a group having the specified number of ring atoms (e.g., C 3-6 The cycloalkyl group may be monovalent or divalent.

[0172] The term "alkoxy" is used in its conventional sense to refer to those alkyl groups that are attached to the remainder of the molecule via an oxygen atom, an amino group, or a sulfur atom, respectively.

[0173] Unless otherwise indicated, the term "halo" or "halogen" by itself or as part of another substituent refers to a fluorine, chlorine, bromine, or iodine atom. In addition, terms such as "haloalkyl" are meant to include monohaloalkyl and polyhaloalkyl. For example, the term "C 1-4 "Haloalkyl" is meant to include trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0174] Unless otherwise stated, the term "aryl" represents a polyunsaturated (usually aromatic) hydrocarbon radical, which can be a monocyclic or polycyclic (maximum three rings) fused together or covalently attached. The term "heteroaryl" refers to an aryl (or ring) containing 1 to 5 heteroatoms selected from N, O, and S, wherein nitrogen and sulfur atoms are optionally oxidized, and nitrogen atoms are optionally quaternized. Heteroaryl can be attached to the remainder of the molecule through heteroatoms. The non-limiting example of aryl includes phenyl, naphthyl, and biphenyl, and the non-limiting example of heteroaryl includes pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, quinolyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl, purinyl, benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl, isobenzofuranyl, isobenzofuranyl, benzothiazolyl ...

[00145] In some embodiments, the aryl and heteroaryl rings may be radicals of the following types: aryl, benzothiazolyl, benzofuranyl, benzothiophenyl, indolyl, quinolinyl, isoquinolinyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl, thiazolyl, furanyl, thienyl, and the like. Substituents for each of the above aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below.

[0175] In some embodiments, the above terms (such as "alkyl," "aryl," and "heteroaryl") will include both substituted and unsubstituted forms of the designated groups. Preferred substituents for each type of group are provided below. For brevity, the terms aryl and heteroaryl will refer to substituted or unsubstituted forms as provided below, while the term "alkyl" and related aliphatic groups refer to the unsubstituted form unless substituted is indicated.

[0176] Substituents for alkyl groups (including those groups commonly referred to as alkylene, alkenyl, alkynyl and cycloalkyl) may be various groups selected from the group consisting of -halogen, -OR', -NR'R", -SR', -SiR'R"R"', -OC(O)R', -C(O)R', -C02R', -CONR'R", -OC(O)NR'R", -NR"C(O)R', -NR'-C(O)NR"R"', -NR"C(O)2R', -NH-C(NH2)=NH,

[0177] -NR'C(NH2)=NH, -NH-C(NH2)=NR', -S(O)R', -S(O)2R', -S(O)2NR'R", -NR'S(O)2R", -CN and -NO2, in a number ranging from zero to (2M'+1), where M' is the total number of carbon atoms in such a group. R', R" and R"' each independently represent hydrogen, unsubstituted C 1-8 Alkyl, unsubstituted heteroalkyl, unsubstituted aryl, aryl substituted with 1-3 halogens, unsubstituted C 1-8 Alkyl, C 1-8 Alkoxy or C 1-8 Thioalkoxy, or unsubstituted aryl-C 1-4 Alkyl. When R' and R" are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring. For example, -NR'R" includes 1-pyrrolidinyl and 4-morpholinyl. The term "acyl" used alone or as part of another group refers to a group in which both substituents on the carbon closest to the point of attachment of the group are replaced with a substituent =0 (e.g., -C(O)CH3, -C(O)CH2CH2OR', etc.).

[0178] Similarly, the substituents for aryl and heteroaryl groups are varied and are typically selected from: -halogen, -OR', -OC(O)R', -NR'R", -SR', -R', -CN, -NO2, -CO2R', -CONR'R", -C(O)R', -OC(O)NR'R", -NR"C(O)R', -NR"C(O)2R', -NR'-C(O)NR"R"', -NH- C(NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2)=NR', -S(O)R', -S(O)2R', -S(O)2NR'R", -NR'S(O)2R", -N3, perfluoro(C1-C4)alkoxy and perfluoro(C1-C4)alkyl, in a number from zero to the total number of open valences on the aromatic ring system; wherein R', R" and R"' are independently selected from hydrogen, C 1-8 Alkyl, C 3-6 Cycloalkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, unsubstituted aryl and heteroaryl, (unsubstituted aryl)-C 1-4 Alkyl and unsubstituted aryloxy-C 1-4 Other suitable substituents include each of the above aryl substituents attached to a ring atom via an alkylene chain of 1 to 4 carbon atoms.

[0179] As used herein, the term "heteroatom" is intended to include oxygen (O), nitrogen (N), sulfur (S), and silicon (Si).

[0180] For the compounds provided herein, a bond from a substituent (typically an R group) to the center of an aromatic ring (e.g., benzene, pyridine, etc.) will be understood to mean a bond providing attachment at any available vertex of the aromatic ring. In some embodiments, this description also includes attachment to a ring fused to an aromatic ring. For example, a bond drawn to the center of an indole benzene moiety will represent a bond attached to any available vertex of a six-membered or five-membered ring moiety of indole.

[0181] As used herein, "a portion derived from..." refers to the portion or fragment of an active substance (e.g., a polypeptide element such as an antibody or a targeting ligand) that remains after the active substance is linked to another portion by certain means (e.g., reacting an active group thereon, or introducing an active group therein for reaction), and the portion or fragment retains the function of the active substance (e.g., the ability to target a desired receptor). Specific linking groups formed by "derivation" include, but are not limited to, -NH-, -CONH-, -CO-, -SS-, and the like.

[0182] The term "pharmaceutically acceptable salt" is intended to include salts of the active compounds prepared with relatively nontoxic acids or bases, depending on the particular substituents on the compounds described herein. When the compounds of the present invention contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of salts derived from pharmaceutically acceptable inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, zinc, and the like. Salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary, and tertiary amines, including substituted amines, cyclic amines, naturally occurring amines, and the like, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydramin, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid, or phosphorous acid, and the like; and salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and the like. Also included are salts of amino acids such as arginine salts, and salts of organic acids such as glucuronic acid or galactunoric acid. Certain specific compounds of the present invention contain both basic and acidic functional groups, which can allow the compounds to be converted into either base or acid addition salts.

[0183] The neutral forms of the compounds can be regenerated by contacting the salt with a base or acid and isolating the parent compound in a conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties (e.g., solubility in polar solvents), but other than that, for the purposes of the present invention, those salts are equivalent to the parent form of the compound.

[0184] In addition to salt forms, the present invention provides compounds in prodrug form. Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present invention. Additionally, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in an ex vivo environment. For example, when placed in a transdermal patch reservoir containing an appropriate enzyme or chemical reagent, a prodrug can be slowly converted to a compound of the present invention.

[0185] Certain compounds of the present invention may exist in unsolvated forms as well as solvated forms, including hydrated forms. Solvated forms are generally equivalent to unsolvated forms and are intended to be encompassed within the scope of this invention. Certain compounds of the present invention may exist in polymorphic or amorphous forms. Generally, for the applications contemplated by the present invention, all physical forms are equivalent and are intended to be encompassed within the scope of this invention.

[0186] Certain compounds of the present invention possess asymmetric carbon atoms (optical centers) or double bonds; racemates, diastereomers, geometric isomers, regioisomers, and individual isomers (e.g., isolated enantiomers) are intended to be encompassed within the scope of the present invention. When compounds provided herein have defined stereochemistry (denoted as R or S, or indicated by dashed or wedge-shaped bonds), those skilled in the art will understand that those compounds are substantially free of other isomers (e.g., at least 80%, 90%, 95%, 98%, 99%, and up to 100% free of other isomers).

[0187] The compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of the isotopic atoms that make up such compounds. An unnatural proportion of an isotope can be defined as the amount of the atom in question found in nature to 100% of that atom. For example, a compound may incorporate a radioactive isotope, such as tritium ( 3 H), iodine-125 ( 125 I) or carbon-14 ( 14 C), or non-radioactive isotopes such as deuterium ( 2 H) or carbon-13 ( 13 C) In addition to those uses described herein, such isotopic variants may provide additional uses. For example, isotopic variants of the compounds of the present invention may have additional uses, including but not limited to, as diagnostic and / or imaging agents, or as cytotoxic / radiotoxic therapeutic agents. Furthermore, isotopic variants of the compounds of the present invention may have altered pharmacokinetic and pharmacodynamic properties, thereby contributing to increased safety, tolerability, or efficacy during treatment. All isotopic variants of the compounds of the present invention, whether or not radioactive, are intended to be encompassed by the present invention.

[0188] Molecular glue degrader

[0189] Current studies have found that the cyclin-dependent kinase (CDK) inhibitor CR8 is also a molecular glue degrader. CR8 can more effectively kill cancer cells by inducing CDK12 / cyclin K to directly form a complex with CUL4 / DDB1, causing cyclin K to be ubiquitinated and degraded through the proteasome system.

[0190] Further structural analysis of the protein-small molecule-protein complex CUL4-RBX1-DDB1-CR8-CDK12-cyclin revealed that CDK12 plays a role similar to that of a CRBN substrate receptor. The presence and correct orientation of the 2-pyridine moiety on the CDK12 surface and CR8 increase the gain of function of CR8, leading to the degradation of cyclin K. Specifically, CR8-phenylpyridine imparts molecular glue activity, inducing cyclin K degradation and increasing the toxicity of CR8.

[0191] By modifying the CR8 structure, the authors discovered that the activity of the CR8 molecular glue is largely dependent on a 2-pyridine moiety exposed on the kinase surface. They concluded that this chemical group enables CR8 to function as a molecular glue degrader. Therefore, chemically modifying the surface-exposed portion of inhibitors can transform them into molecular glue degraders for specific protein targets.

[0192] However, the inventors have found in their research that the compounds having the structures shown in the present invention, formula (I), formula (II), formula (II-1), and formula (II-2) still have excellent effects of inducing the degradation of cell cycle proteins such as cyclin K, regardless of the presence or absence of a pyridine substituent (or other nitrogen-containing heteroaromatic ring substituent) or whether they are pyridine substituents (or other nitrogen-containing heteroaromatic ring substituents). Based on this, the inventors have provided a series of novel cell cycle protein regulators (more specifically, molecular glue degraders).

[0193] In one aspect, the present invention provides a molecular glue degrader (also referred to herein as a degrader or a cell cycle protein degrader), wherein the molecular glue degrader is a compound as shown in formula (I) or a pharmaceutically acceptable salt thereof,

[0194] Wherein, each group is as defined in the first aspect.

[0195] In a preferred embodiment, the molecular glue degradation agent is a compound represented by formula (II), formula (II-1) or formula (II-2) or a pharmaceutically acceptable salt thereof;

[0196] Wherein, each group is as defined above.

[0197] In one aspect of the present invention, a molecular glue degradation agent is provided as shown below:

[0198] Wherein, n2 is 2-4 (i.e., the ring where n2 is located is a 4- to 6-membered ring), and ring Ar 1 Optionally, n3 R a Substituent substitution (not shown) and ring Cr 1 Also optionally n4 R b Substituents (not shown); R 1 、R 2 、R 3 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 8 、X 9 、Subscript n1、n3、n4、Ar 1 Cr 1 、R a and R b As defined elsewhere in this document.

[0199] In another aspect of the present invention, a molecular glue degradation agent is provided as shown below:

[0200] Wherein, n2 is 2-4 (i.e., the ring where n2 is located is a 4- to 6-membered ring); Ring Ar 1 Optionally, n3 R a Substituents (not shown) and ring Ar 2 Also optionally n4 R b Substituents (not shown); R 1 、R 2 、R 3 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 8 、X 9 、Subscript n1、n3、n4、Ar 1 Cr 1 、R a and R b As defined elsewhere in this document.

[0201] In another preferred embodiment, the compound is selected from Table A.

[0202] In another preferred embodiment, the compound is selected from the following Table B1 or Table B2;

[0203] Table B1

[0204] Table B2

[0205] Conjugate

[0206] The present invention also provides a conjugate, which is a compound comprising a molecular glue degrading agent portion or a pharmaceutically acceptable salt thereof, wherein the molecular glue degrading agent portion is derived from the molecular glue degrading agent as defined above.

[0207] In another preferred embodiment, the conjugate further comprises a polypeptide element or a targeting ligand portion.

[0208] In another preferred embodiment, the conjugate is a conjugate formed by linking the compound described in the first aspect with a polypeptide element or a targeting ligand.

[0209] polypeptide element

[0210] As used herein, the term "polypeptide element" includes peptide segments (e.g., short peptides of 3-20 aa) or proteins. In addition, the term also includes intact proteins or fragments thereof. Preferred polypeptide elements include antibodies (e.g., intact antibodies, single-chain antibodies, nanobodies, antibody fragments), especially antibodies against tumor cell markers (e.g., tumor markers located on the surface of tumor cells, such as cell surface receptors) or against inflammatory factors (e.g., inflammatory factors associated with autoimmune diseases).

[0211] As used herein, the terms "antibody" or "immunoglobulin" are heterotetrameric glycoproteins of approximately 150,000 daltons with identical structural features, consisting of two identical light chains (L) and two identical heavy chains (H). Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide bonds varies between heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bonds. Each heavy chain has a variable region (VH) at one end, followed by multiple constant regions. Each light chain has a variable region (VL) at one end and a constant region at the other end; the constant region of the light chain is opposite the first constant region of the heavy chain, and the variable region of the light chain is opposite the variable region of the heavy chain. Specific amino acid residues form an interface between the variable regions of the light and heavy chains.

[0212] As used herein, the terms "single-domain antibody" and "nanobody" have synonymous meanings and refer to the construction of a single-domain antibody consisting solely of a single heavy chain variable region by cloning the variable region of an antibody heavy chain. This is the smallest fully functional antigen-binding fragment. Typically, antibodies naturally lacking the light chain and heavy chain constant region 1 (CH1) are first obtained, and then the variable region of the antibody heavy chain is cloned to construct a single-domain antibody consisting solely of a single heavy chain variable region.

[0213] As used herein, the term "variable" refers to certain portions of the variable region of an antibody that differ in sequence, which contribute to the binding and specificity of each particular antibody for its specific antigen. However, variability is not evenly distributed throughout the variable region of an antibody. It is concentrated in three segments called complementarity determining regions (CDRs) or hypervariable regions in the variable regions of the light and heavy chains. The more conserved portions of the variable region are called framework regions (FRs). The variable regions of native heavy and light chains each contain four FR regions, which are generally in a β-pleated configuration and are connected by three CDRs that form a connecting loop, which in some cases can form a partially folded structure. The CDRs in each chain are closely together through the FR regions and, together with the CDRs of the other chain, form the antigen-binding site of the antibody. The constant regions are not directly involved in the binding of the antibody to the antigen, but they exhibit different effector functions, such as involvement in the antibody's antibody-dependent cellular toxicity.

[0214] The "light chains" of vertebrate antibodies (immunoglobulins) can be assigned to one of two distinct classes, called kappa and lambda, based on the amino acid sequence of their constant regions. Immunoglobulins can be divided into different classes based on the amino acid sequence of their heavy chain constant regions. There are five main classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, some of which are further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA, and IgA2. The heavy chain constant regions corresponding to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional configurations of the different classes of immunoglobulins are well known in the art.

[0215] Generally, an antibody's antigen-binding properties are described by three specific regions located in the variable regions of the heavy and light chains, known as the variable regions (CDRs). These regions are divided into four framework regions (FRs). The amino acid sequences of the four FRs are relatively conserved and do not directly participate in the binding reaction. These CDRs form a ring structure, spatially close to each other through the β-sheet formed by the FRs between them. The CDRs on the heavy chain and the corresponding CDRs on the light chain constitute the antibody's antigen-binding site. The amino acid sequences of antibodies of the same type can be compared to determine which amino acids constitute the FR or CDR regions.

[0216] In the present invention, polypeptide elements may include not only intact antibodies, but also immunologically active antibody fragments (e.g., Fab or (Fab')2 fragments; antibody heavy chains; or antibody light chains) or fusion proteins formed by antibodies and other sequences. Therefore, the present invention also includes fragments, derivatives, and analogs of such antibodies.

[0217] Targeting ligands

[0218] A targeting ligand (or target protein moiety or target protein ligand or ligand) is a small molecule that is capable of binding to a target protein of interest.

[0219] In some embodiments of the present application, the targeting ligand may be (or be derived from) a target molecule.

[0220] Some embodiments of the present application relate to target molecules, representative target molecules include but are not limited to: folic acid, Hsp90 inhibitors, kinase inhibitors, MDM2 inhibitors, compounds targeting proteins containing human BET bromodomains, compounds targeting cytoplasmic signaling protein FKBP12, HDAC inhibitors, human lysine methyltransferase inhibitors, angiogenesis inhibitors, immunosuppressive compounds, compounds targeting aryl hydrocarbon receptor (AHR) and compounds targeting tumor anaerobic microenvironment.

[0221] In certain embodiments, the targeting ligand is capable of binding to a kinase, a BET bromodomain-containing protein, a cytoplasmic signaling protein (e.g., FKBP12), a nuclear protein, a histone deacetylase, a lysine methyltransferase, a protein that regulates angiogenesis, a protein that regulates the immune response, the aryl hydrocarbon receptor (AHR), an estrogen receptor, an androgen receptor, a glucocorticoid receptor, or a transcription factor (e.g., SMARCA4, SMARCA2, TRIM24).

[0222] In certain embodiments, the kinases to which the targeting ligand can bind include, but are not limited to, tyrosine kinases (e.g., AATK, ABL, ABL2, ALK, AXL, BLK, BMX, BTK, CSF1R, CSK, DDR1, DDR2, EGFR, EPHA1, EPHA2, EPHA3, EPHA4, EPHA5, EPHA6, EPHA7, EPHA8, EPHA10, EPHB1, EPHB2, EPHB3, EPHB4, EPHB6, ERBB2, ERBB3, ERBB4, FER, FES, FGFR1, FGFR2, FGFR3, FGFR4, FGR, FLT1, FLT3, FLT4, FRK, FYN, GSG2, HCK, HRAS, HSP90, IGF1R, ILK, INSR, INSRR, IRAK4, ITK, JAK1, JAK2, JAK3, KDR, KIT, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD, KD RAS, KSP, KSR1, LCK, LMTK2, LMTK3, LTK, LYN, MATK, MERTK, MET, MLTK, MST1R, MUSK, NPR1, N RA S, NTRK1, NTRK2, NTRK3, PDGF RA , PDGF RB , PLK4, PTK2, PTK2B, PTK6, PTK7, RET, ROR1, ROR2, ROS1, RYK, SGK493, SRC, SRMS, STYK1, SYK, TEC, TEK, TEX14, TIE1, TNK1, TNK2, TNNI3K, TXK, TYK2, TYRO3, YES1, or ZAP70), serine / threonine kinases (e.g., casein kinase 2, protein kinase A, protein kinase B, protein kinase C, Raf kinase, CaM kinase, AKT1, AKT2, AKT3, ALK1, ALK2, ALK3, ALK4, AuroraA, AuroraB, AuroraC, CHK1, CHK2, CLK1, CLK2, CLK3, DAPK1, DAPK2, DAPK3, DMPK, ER , TSSK1, TSSK2, MLK1, or MLK2), cyclin-dependent protein kinases (e.g., Cdk1-Cdk11), and leucine-rich repeat kinases (e.g., LRRK2).

[0223] In another preferred embodiment, the conjugate is a compound as shown in Formula III or a pharmaceutically acceptable salt thereof; M D -M CL -M CP (III)

[0224] Among them, M D It is the molecular glue degradation agent part; M CL No or connector part; M CP is a portion derived from a polypeptide element or a targeting ligand;

[0225] In another preferred embodiment, the targeting ligand refers to a small molecule that can bind to an extracellular receptor.

[0226] In another preferred embodiment, the polypeptide element includes (but is not limited to): polypeptide, antibody, antibody fragment, fusion protein, or a combination thereof.

[0227] In another preferred embodiment, M CP Selected from the group consisting of a polypeptide, an antibody, an antibody fragment, a fusion protein, or a small molecule ligand portion capable of binding to an extracellular receptor.

[0228] In another preferred embodiment, the antibody includes (but is not limited to): nanobody, small molecule antibody (minibody), antibody fragment (such as scFv, Fab), dibody, monoclonal antibody (mAb), or a combination thereof.

[0229] In another preferred embodiment, the target of the polypeptide (such as a targeting polypeptide) includes but is not limited to: EGFR, FGFR, SSTR1-14, GnRH, TRPV1-6, RGD, iRGD, EphA2, or a combination thereof.

[0230] In another preferred embodiment, the targets that the small molecule ligand can bind to include (but are not limited to): FR, HSP90, PSMA, ASGPR, and combinations thereof.

[0231] In another preferred embodiment, the antibody can bind to an antigen or receptor selected from the following group (for example, one (i.e., monofunctional antibody) or two (i.e., bifunctional antibody) or more (i.e., multifunctional antibody) antigens and / or receptors selected from the following group): DLL3, EDAR, CLL1, BMPR1B, E16, STEAP1, 0772P, MPF, 5T4, NaPi2b, Sema 5b, PSCA hlg, ETBR, MSG783, STEAP2, TrpM4, CRIPTO, CD21, CD22, CD79b, CD19, CD37, CD38, CD138, FcRH2, B7-H4, HER2, NCA, MDP, IL20Rα, Brevican, EphB2R, ASLG659, PSCA, GEDA, BAFF-R, CD79a, CXCR5, HLA-DOB, P2X5, CD72, LY64, FcRH1, IRTA2, TENB2, PMEL17, TMEFF1, GDNF-Ra1, Ly6E, TMEM46, Ly6G6D, LGR5, RET, LY6K, GPR19, GPR54, ASPHD1, Tyrosinase, TM EM118, GPR172A, MUC1, CD70, CD71, MUC16, methotrexate, FOLR1, TroP1-2, gpNMB, EGFR, ENPP3, PSMA, CA6, GPC-3, PTK7, CD44, CD56, TIM-1, Cadherin-6, ASG-15ME, ASG-22ME, CanAg, AXL, CEACAM5, EphA4, cMet, FGFR2, FGFR3, CD123, Her3, LAMP1, LRRC15, TDGF1, CD66, CD25, BCMA, GCC, Noch3, cMet, EGFR and CD33, or receptors such as CD70, Trop2, PD-L1, CD47, CLDN-18.2.

[0232] In another preferred embodiment, the targeting ligand can also bind to receptors that can be targeted by specific small molecules, such as folic acid, HSP90, glucose transporter 1 (GLUT1), aminopeptidase N (APN), low-density lipoprotein receptor-related protein 1 (LRP1), prostate-specific membrane antigen (PSMA), integrin αvβ3, bombesin receptor, somatostatin receptor (SSTR), tumor hypoxic microenvironment, and carbonic anhydrase IX (CAIX) and other receptors.

[0233] Prodrug

[0234] The present invention also provides a prodrug, which is a compound comprising a molecular glue degrading agent portion or a pharmaceutically acceptable salt thereof, wherein the molecular glue degrading agent portion is derived from the molecular glue degrading agent as defined above.

[0235] In another preferred embodiment, the prodrug further comprises a leaving moiety.

[0236] In another preferred embodiment, the prodrug is a compound represented by Formula IV or a pharmaceutically acceptable salt thereof;

[0237] The prodrug is a compound represented by Formula IV or a pharmaceutically acceptable salt thereof;

[0238] MD-MPL-MLG(IV) where M D It is the molecular glue degradation agent part; M PL Is none or a connecting group; M LG For the leaving part.

[0239] In another preferred embodiment, the leaving portion is capable of leaving and releasing the molecular glue degrading agent when it is located at or reaches the target site (such as the tumor microenvironment).

[0240] In another preferred embodiment, the leaving portion comprises an enzyme recognition fragment and optionally one or more hydrophilic groups.

[0241] In another preferred embodiment, M PL is none or can be combined with M LG A group that forms a cleavable bond (e.g., -C(O)-).

[0242] In another preferred embodiment, M LG for

[0243] Among them, M LG is optionally substituted C 1-4 Alkylene (preferably, methylene); R LG represents one or more groups containing hydrophilic groups and / or enzyme recognition fragments.

[0244] In another preferred embodiment, the enzyme recognition fragment is selected from the following group:

[0245] In another preferred embodiment, the hydrophilic group includes (but is not limited to) one or more of the following: hydroxyl, -(CH2CH2O) 1-10 -、-SO3H、-PO3H2、-COOH.

[0246] In another preferred embodiment, the leaving part is selected from the following Table C1:

[0247] Table C1

[0248] In another preferred embodiment, M D As shown in Formula Ia

[0249] in,

[0250] M 3a No (single bond), R 3a 、-NH-;

[0251] R 3a Selected from the group consisting of: -O-, optionally substituted -OC 1-6- Alkylene-, optionally substituted-C(O)-C 1-6 Alkylene-, optionally substituted-C(O)-NH-, optionally substituted-C(O)-N(C 1-6- alkyl)-, optionally substituted-C 1-6 Haloalkylene-;

[0252] R 1 、R 2 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 7 、X 8 、X 9 、X 10 、X 11 、X 12 、M 1 、M 2, subscript n1, subscript n3, subscript n4, Ar 1 Cr 1 、R a and R b As defined in Formula I.

[0253] In another preferred embodiment, M D As shown in formula II-a

[0254] Among them, M 3a As defined in Formula Ia; R 2 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、M 1 and M 2 and M 3a As defined in Formula II.

[0255] In another preferred embodiment, M D As shown in formula II-1a

[0256] Among them, R 2 、X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、X 9 、X 10 and X 11 As defined in Formula II-1.

[0257] In another preferred embodiment, M D 、M PL and M GL Each is independently the corresponding group in the specific compound shown in Table C.

[0258] In another preferred embodiment, the conjugate is selected from Table C

[0259] Table C

[0260] Active ingredient

[0261] As used herein, the term "compound of the present invention" refers to a compound as defined in the first aspect (e.g., a compound of Formula I, Formula II, Formula II-1, or Formula II-2). The term also includes various crystalline forms or pharmaceutically acceptable salts of the compound of Formula I. In this article, the active ingredient may also be a conjugate formed by the compound and an antibody or polypeptide (e.g., a conjugate defined in the seventh aspect).

[0262] Pharmaceutical compositions and methods of administration

[0263] Because the compounds of the present invention have excellent activity in inducing cyclin K degradation, the compounds of the present invention and their various crystalline forms, pharmaceutically acceptable inorganic or organic salts, hydrates, or solvates, as well as pharmaceutical compositions containing the compounds of the present invention as the primary active ingredient, or conjugates formed from the compounds of the present invention and antibodies or polypeptides, can be used to treat or prevent diseases related to or involving cyclin K. According to prior art, the compounds of the present invention can be used to treat the following diseases: cancer, etc.

[0264] The pharmaceutical composition of the present invention comprises a safe and effective amount of a compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. "Safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects.

[0265] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0266] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration routes include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0267] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0268] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0269] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0270] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0271] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0272] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0273] Dosage forms for topical administration of the compounds of this invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0274] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0275] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage. Since the molecular glue degrader of the present invention has a more efficient tumor-killing effect than inhibitors, the degrader of the present invention only requires a low dose to achieve the effect of treating or preventing tumors compared to the use of inhibitors. Generally, for a person weighing 60 kg, the daily dosage is usually 0.01 to 10 mg (calculated as the degrader or the degrader portion of the conjugate or prodrug), preferably 0.05 to 5 mg, and more preferably 0.1-1 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health status, which are all within the skill of a skilled physician.

[0276] The main advantages of the present invention include

[0277] (a) The compound of the present invention has an excellent effect of inducing the degradation of cyclin K.

[0278] (b) Further studies have revealed that the compounds of the present invention also have the function of inducing the degradation of other cell cycle proteins, thereby further increasing their cytotoxicity.

[0279] (c) Inhibitors typically require high doses to inhibit the function of pathogenic proteins and achieve the goal of suppressing tumor growth. Unlike conventional inhibitors, the degraders provided by the present invention typically require only a catalytic dose to directly degrade the pathogenic target protein and prevent the development of mutational resistance. Clinically, the dosage of inhibitors typically ranges from several milligrams to several hundred milligrams, while molecular glue degraders can be used in amounts of less than 1 milligram.

[0280] (d) Since the anti-tumor therapeutic effect can be exerted at a lower dose, the compound of the present application has a higher therapeutic window.

[0281] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which no specific conditions are specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0282] A. Preparation Examples

[0283] General synthetic method

[0284] The compounds of the present invention can be prepared, isolated or obtained by any method that is obvious to those skilled in the art. The compounds of the present invention can also be prepared according to the exemplary preparation schemes provided below (such as the method in the examples). The reaction conditions, steps and reactants not provided in the exemplary preparation schemes are obvious and known to those skilled in the art. As used herein, the symbols and conventions used in these processes, schemes and examples, regardless of whether specific abbreviations are specifically defined, have meanings well known to those skilled in the art. Specifically, but not limited to, the following abbreviations may be used in the examples and throughout the specification: g (gram); mg (milligram); mL (milliliter); μL (microliter); mm (millimole); μM (micromolar); MHz (hertz); MHz (megahertz); mmol (millimole); hr or hrs (hours); min (minutes); MS (mass spectrometry); ESI (electrospray ionization); TLC (thin layer chromatography); HPLC (high performance liquid chromatography); THF (tetrahydrofuran); CDCl3 (deuterated chloroform); AcOH (acetic acid); DCM (dichloromethane); DMSO (dimethyl sulfoxide); EtOAc (ethyl acetate); MeOH (methanol); ; and BOC (tert-butyloxycarbonyl), etc.

[0285] Unless otherwise stated, the starting materials used in the examples were commercially available or synthesized in a manner known to those skilled in the art or in a manner analogous to that described in the examples.

[0286] Example 1: Synthesis of Compound UB2-001

[0287] Step 1: UB2-001: (S)-N-([1,1'-biphenyl]-4-ylmethyl)-9-isopropyl-2-(3-(trifluoromethyl)piperazin-1-yl)-9H-purin-6-amine

[0288] Compound UB2-001a (84 mg, 0.22 mmol), UB2-001b (140 mg, 0.91 mmol), and DIPEA (2 mL) were dissolved in NMP (4 mL). The reaction was stirred in a microwave oven at 180°C for 8 hours. After completion, the reaction solution was cooled to room temperature. Water was added and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried, filtered, and purified on a silica gel column (DCM / MeOH = 10 / 1) and lyophilized to obtain the target compound UB2-001 (69.77 mg, 64% yield) as a light yellow solid. LCMS [M+1] + =496.3. 1 H NMR (400MHz, CDCl3) δ7.62-7.49(m,5H),7.49-7.37(m,4H),7.37-7.31(m,1H),6.21(s,1H),4.99-4.73(m,3H) ,4.74-4.60(m,2H),3.41-3.26(m,1H),3.17-2.96(m,3H),2.85(td,J=11.3,3.2Hz,1H),1.55(d,J=6.8Hz,6H).

[0289] Example 2: Synthesis of Compound UB2-002

[0290] Step 1: UB2-002: (R)-N-([1,1'-biphenyl]-4-ylmethyl)-9-isopropyl-2-(3-(trifluoromethyl)piperazin-1-yl)-9H-purin-6-amine

[0291] Compound UB2-002a (58 mg, 0.15 mmol), UB2-002b (94 mg, 0.61 mmol), and DIPEA (1.5 mL) were dissolved in NMP (3 mL). The reaction was stirred in a microwave oven at 180°C for 10 hours. After completion, the reaction solution was cooled to room temperature. Water was added and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried, filtered, and purified on a silica gel column (DCM / MeOH = 10 / 1) and lyophilized to yield the target compound UB2-002 (42 mg, 55% yield) as a white solid. LCMS [M+1] + =496.3

[0292] 1H NMR (400MHz, CDCl3) δ7.65-7.50(m,5H),7.50-7.37(m,4H),7.37-7.30(m,1H),6.22(s,1H),4.99-4.72(m,3H),4.72-4. 56(m,2H),3.41-3.25(m,1H),3.05(ddd,J=23.0,18.0,11.0Hz,3H),2.85(td,J=11.3,3.1Hz,1H),1.56(d,J=6.8Hz,6H).

[0293] Example 3: General method for synthesizing compound UB2-003

[0294] Step 1: UB2-003: 4-(6-(([1,1'-biphenyl]-4-ylmethyl)amino)-9-isopropyl-9H-purin-2-yl)piperazin-2-one

[0295] Compound UB2-003a (60 mg, 0.16 mmol), 2-piperazinone (79.58 mg, 0.80 mmol), DIPEA (164.55 mg, 1.27 mmol), and N-methylpyrrolidone (5 mL) were added to a microwave tube and heated at 180°C for 8 hours. The mixture was cooled to room temperature, concentrated under reduced pressure, and purified by silica gel column chromatography (12 g, 40 mL / min; DCM: (10% MeOH in DCM) = 0-33%) to obtain the crude product as a yellow liquid. The crude product was further purified by preparative liquid chromatography (mobile phase: formic acid / water / acetonitrile) to obtain the product UB2-003 (33.43 mg, yield: 47.6%) as a white solid. LCMS [M+1] + =442.2

[0296] 1 H NMR (400MHz, DMSO-d6) δ8.15(s,1H),7.96(s,1H),7.89(s,1H),7.60(dd,J=15.0,7.8Hz,4H),7.49-7.39(m,4H),7.33(t,J =7.3Hz,1H),4.59(ddd,J=20.1,13.6,6.7Hz,3H),4.15(s,2H),3.85(t,J=5.2Hz,2H),3.21(s,2H),1.48(d,J=6.8Hz,6H).

[0297] Example 4: Synthesis of Compound UB2-004

[0298] Step 1: UB2-004b

[0299] Compound UB2-004a (500 mg, 2.05 mmol) was dissolved in ammonia-methanol solution (12 mL) and reacted at 100°C for 72 hours. The reaction solution was dried by rotary evaporation, water was added, and extraction was performed with ethyl acetate (30 mL x 3). The organic layer was washed with brine, dried over anhydrous sodium sulfate, and dried by rotary evaporation. Purification by column chromatography (dichloromethane / methanol = 10 / 1) afforded compound UB2-004b (310 mg, 66% yield) as a yellow solid. LCMS [M+1] + =230.1

[0300] Step 2: UB2-004c

[0301] Compound UB2-004b (310 mg, 1.36 mmol) was dissolved in dichloromethane (6 mL), and a solution of hydrogen chloride in dioxane (3 mL, 4 M) was added. The mixture was stirred at room temperature for 2 hours. The reaction mixture was dried to afford compound UB2-004c (300 mg, 100% yield) as a pale yellow solid, which was used directly in the next reaction. LCMS [M+1] + =130.1

[0302] Step 3: UB2-004

[0303] Compound UB2-004c (300 mg, 1.36 mmol) and compound UB2-004d (150 mg, 0.40 mmol) were dissolved in 8 mL of N-methylpyrrolidone, and N,N-diisopropylethylamine (4 mL) was added and microwave-treated at 180°C for 10 hours. The reaction solution was cooled to 20°C, quenched with water (20 mL), and extracted three times with ethyl acetate (20 mL). The organic layer was washed with brine, dried over anhydrous sodium sulfate, and spin-dried. Preparative chromatography was then performed to obtain compound UB2-004 (22.79 mg, 13% yield) as a white solid. LCMS [M+1] + =471.3. 1H NMR (400MHz, DMSO-d6) δ8.03 (s, 1H), 7.86 (s, 1H), 7.64-7.59 (m, 2H), 7.57 (d, J = 8.2Hz, 2H),7.46(d,J=7.0Hz,2H),7.43(d,J=7.8Hz,2H),7.33(t,J=7.3Hz,1H),7.27(s,1H),7 .13(s,1H),4.71-4.50(m,4H),4.33(d,J=12.6Hz,1H),3.14(dd,J=10.0,3.3Hz,1H),2. 92(d,J=12.1Hz,1H),2.85-2.73(m,2H),2.60(t,J=10.2Hz,1H),1.46(d,J=6.7Hz,6H).

[0304] Example 5: General method for synthesizing compound UB2-006

[0305] Step 1: UB2-006: 1-(6-(([1,1'-biphenyl]-4-ylmethyl)amino)-9-isopropyl-9H-purin-2-yl)piperidin-4-ol

[0306] Compound UB2-006a (60 mg, 0.16 mmol), 4-hydroxypiperidine (80.49 mg, 0.80 mmol), DIPEA (164.55 mg, 1.27 mmol), and N-methylpyrrolidone (5 mL) were added to a microwave tube and heated at 180°C for 3 hours. After concentration under reduced pressure, the mixture was purified by silica gel column chromatography (12 g, DCM:MeOH = 10:1) to obtain the crude product. Further purification by preparative liquid chromatography (mobile phase: formic acid / water / acetonitrile) afforded the product UB2-006 (36.6 mg, 52% yield) as a white solid. LCMS [M+1] + =443.3. 1 H NMR (400MHz, DMSO-d6) δ7.97(s,1H),7.83(s,1H),7.62(d,J=7.4Hz,2H),7.57(d,J=8.1Hz,2H),7.44(t,J=7.9Hz,4H),7.33(t,J=7.3Hz,1H),4.78-4 .47(m,4H),4.31(d,J=13.2Hz,2H),3.69-3.59(m,1H),3.08(dd,J=16.8,6 .4Hz,2H),1.71(d,J=9.8Hz,2H),1.46(d,J=6.7Hz,6H),1.29-1.21(m,2H).

[0307] Example 6: General method for synthesizing compound UB2-007

[0308] Step 1: UB2-007: (1r,4r)-4-((6-(([1,1′-biphenyl]-4-ylmethyl)amino)-9-isopropyl-9H-purin-2-yl)amino)cyclohexanol

[0309] Compound UB2-007a (60 mg, 0.16 mmol), trans-4-aminocyclohexanol (91.65 mg, 0.80 mmol), DIPEA (164.55 mg, 1.27 mmol), and N-methylpyrrolidone (5 mL) were added to a microwave tube and heated at 180°C for 3 hours. The mixture was cooled to room temperature, concentrated under reduced pressure, and purified by silica gel column chromatography (12 g, DCM:MeOH = 10:1) to obtain a crude product. This was further purified by preparative liquid chromatography (mobile phase: formic acid / water / acetonitrile) to obtain the product UB2-007 (29.4 mg, yield: 40.51%) as a white solid. LCMS [M+1] + =457.3. 1 H NMR (400MHz, DMSO-d6) δ7.89(s,1H),7.78(s,1H),7.62(d,J=7.4Hz,2H),7.57(d,J=8.1Hz,2H),7.44(t,J=7.3Hz,4H),7.33(t,J =7.3Hz,1H),6.03(d,J=7.4Hz,1H),4.81-4.37(m,4H),3.60(s,1H),1.95-1.67(m,4H),1.47(t,J=7.7Hz,6H),1.28-1.10(m,4H).

[0310] Example 7: General method for synthesizing compound UB2-011

[0311] Step 1: UB2-011b: 5-Isopropyl-4-nitro-1H-pyrazole-3-carboxylic acid

[0312] Dissolve UB2-011a (10 g, 64.863 mmol) in concentrated sulfuric acid. Add concentrated nitric acid (15 mL) dropwise under an ice bath. Heat to 100°C and stir for 4 hours. Pour the reaction solution into ice water. A white solid precipitates. Filter and dry the filter cake to obtain a white solid product (UB2-011b, 10 g, 77.4% yield). LCMS [M+1] + =200.1

[0313] Step 2: UB2-011c: 5-Isopropyl-4-nitro-1H-pyrazole-3-carboxamide

[0314] UB2-011b (10 g, 54.225 mmol) was dissolved in SOCl2 (60 mL), heated to 80°C and stirred for 3 hours, then the solvent was removed. The intermediate acid chloride was dissolved in THF (100 mL), and then NH4OH (20 mL) was added under ice bath and stirred for 1 hour. The reaction mixture was concentrated in vacuo to give the crude product (UB2-011c, 10 g, 100% yield) as a white solid. LCMS [M+1] + =199.1

[0315] Step 3: UB2-011d: 4-amino-5-isopropyl-1H-pyrazole-3-carboxamide

[0316] To a solution of UB2-011c (10 g, 50.201 mmol) in methanol (100 mL) was added Pd / C (1 g, 10% wt). The mixture was heated to 50°C under a hydrogen atmosphere and stirred for 18 hours. The mixture was filtered under reduced pressure, and the filtrate was concentrated in vacuo to give a white solid (UB2-011d, 10 g, 100% yield). LCMS [M+1] + =169.1

[0317] Step 4: UB2-011e: 3-Isopropyl-2,4-dihydro-5H-pyrazolo[4,3-d]pyrimidine-5,7(6H)-dione

[0318] Urea (1.5 g, 25 mmol) was added to UB2-011d (1.4 g, 8.32 mmol). The mixture was heated to 180°C and stirred for 1 hour. The mixture was slowly diluted with ice water (20 mL), extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo and purified by flash chromatography to obtain the desired compound as a brown solid (UB2-011e, 790 mg, 48.6% yield). LCMS [M+1] + =195.1

[0319] Step 5: UB2-011f: 5,7-dichloro-3-isopropyl-2H-pyrazolo[4,3-d]pyrimidine

[0320] UB2-011e (740 mg, 3.811 mmol) was dissolved in POCl3 solution (10 mL), heated to 100°C and stirred for 18 hours. The solvent was removed and the mixture was purified by flash chromatography to obtain a white solid (UB2-011f, 190 mg, 21.57% yield). LCMS [M+1] + =231.1

[0321] Step 6: UB2-011h: N-([1,1'-biphenyl]-4-ylmethyl)-5-chloro-3-isopropyl-2H-pyrazolo[4,3-d]pyrimidin-7-amine

[0322] To a solution of UB2-011f (190 mg, 0.822 mmol) in n-BuOH (5 mL) was added UB2-011g (180 mg, 0.983 mmol) and DIPEA (160 mg, 1.233 mmol). The mixture was heated to 120°C and stirred for 1 hour. The solvent was removed and the sample was directly purified by flash chromatography to afford a yellow solid (UB2-011h, 194 mg, 62.4% yield). LCMS [M+1] + =378.1

[0323] Step 7: UB2-011i: tert-Butyl-7-([1,1'-biphenyl]-4-ylmethyl)(tert-butoxycarbonyl)amino)-5-chloro-3-isopropyl-2H-pyrazolo[4,3-d]pyrimidine-2-carboxylate

[0324] To a solution of UB2-011h (440 mg, 1.16 mmol) in tetrahydrofuran (30 mL) was added butyl dicarbonate (3.05 g, 14 mmol), N,N-diisopropylethylamine (1.5 g, 11.6 mmol), and 4-dimethylaminopyridine (566.8 mg, 4.64 mmol). The reaction mixture was heated to 50°C and stirred overnight. The reaction was complete, as determined by LCMS [M+H]. + =578.2, post-treatment: the reaction solution was poured into water and extracted three times with ethyl acetate (3*50 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and pulled dry. The crude product was purified by column chromatography (eluent: ethyl acetate / petroleum ether = 0-5%) to obtain the target compound (UB2-011i, 503 mg, yield 75%) as a white solid. LCMS [M+1] + =578.2

[0325] Step 8: UB2-011k: tert-Butyl-(S)-7-([1,1'-biphenyl]-4-ylmethyl)(tert-butoxycarbonyl)amino)-5-(4-(tert-butoxycarbonyl)-2-(hydroxymethyl)piperazin-1-yl)-3-isopropyl-2H-pyrazolo[4,3-d]pyrimidine-2-carboxylate

[0326] To a 100 mL three-necked flask was added UB2-011i (403.6 mg, 0.7 mmol), UB2-011j (453 mg, 2.1 mmol), tris(dibenzylideneacetone)dipalladium (256.4 mg, 0.28 mmol), (±)-2,2'-bis-(diphenylphosphino)-1,1'-binaphthyl (348.7 mg, 0.56 mmol), cesium carbonate (684 mg, 2.1 mmol), and dioxane (40 mL). The reaction mixture was purged with argon several times, heated to 100°C, and stirred overnight. The reaction was complete as determined by LCMS [M+H]. + =758.4, post-treatment: the reaction solution was poured into water and extracted three times with ethyl acetate (3*50 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and pulled dry. The crude product was passed through a column chromatography [eluent: ethyl acetate / petroleum ether = 0-15%] to obtain the target compound (UB2-011k, ​​170 mg, yield 32%) as a yellow solid. LCMS [M+1] + =758.4

[0327] Step 9: UB2-011: (S)-(1-(7-(([1,1'-biphenyl]-4-ylmethyl)amino)-3-isopropyl-2H-pyrazolo[4,3-d]pyrimidin-5-yl)piperazin-2-yl)methanol

[0328] To a solution of UB2-011k (170 mg, 0.35 mmol) in dichloromethane (15 mL) was added dropwise a solution of hydrochloric acid-dioxane (7.5 mL, 4 mol / L). The reaction was stirred at 30°C overnight. The reaction was complete as determined by LCMS [M+H] + =458, post-processing: the reaction solution was directly drained, a small amount of ethyl acetate was added to slurry, filtered, and the filter cake was prepared (formic acid) and purified to obtain the target compound (UB2-011, 11.77 mg, yield 7.35%) as a white solid, LCMS [M+1] + =458.3.

[0329] 1H NMR(400MHz,DMSO-d6)δ12.15(s,1H),8.26(s,1H),7.67-7.58(m,4H),7.54-7.39( m,4H),7.38-7.32(m,1H),4.76-4.61(m,3H),4.47(d,J=13.6Hz,1H),3.78(t,J=9. 3Hz,1H),3.48-3.45(m,1H),3.30(s,1H),3.17(q,J=6.9Hz,1H),3.00(q,J=12.9,1 2.1Hz,2H),2.78(dd,J=12.3,4.3Hz,1H),2.71-2.63(m,1H),1.32(d,J=6.9Hz,6H).

[0330] Example 8: General method for synthesizing compound UB2-012

[0331] Step 1: UB2-012c: Ethyl N-[(4-isopropyl-1H-pyrazol-5-yl)aminomethylthio]carbamate

[0332] UB2-012a (4 g, 31.95 mmol) was dissolved in DCM (100 mL), and UB2-012b (4.61 g, 35.144 mmol) was added to the reaction mixture. The reaction mixture was stirred at 20°C for 18 hours. After concentration, the mixture was slurried with PE / EtOAc (10 / 1, 50 mL) and dried to obtain a yellow solid compound UB2-012c (5.5 g, 67% yield). [M+1] + =257.1.

[0333] Step 2: UB2-012d: 8-Isopropyl-2-thioloxy-2,3-dihydropyrazolo[1,5-a][1,3,5]triazin-4(1H)-one

[0334] To a mixture of UB2-012c (5.5 g, 21.5 mmol) in acetonitrile (100 mL) was added K2CO3 (7.43 g, 53.75 mmol). The reaction was stirred at 80°C for 18 hours. The reaction was quenched with water and the pH was adjusted to 7-8 with acetic acid. The solvent was drained and washed with water (500 mL). Filtered to afford UB2-012d (3.99 g, 88% yield) as a white solid. LCMS [M+1] + =211.1.

[0335] Step 3: UB2-012e: 2-Bromo-8-isopropylpyrazolo[1,5-a][1,3,5]triazin-4(3H)-one

[0336] UB2-012d (1000 mg, 4.762 mmol, 1 eq) was dissolved in 48% HBr (80 mL) and cooled to 0°C. A solution of Br2 (0.5 mL, 2 eq) in 48% HBr (20 mL) was then added dropwise to the reaction mixture. The reaction mixture was stirred at 0°C for half an hour. The reaction mixture was poured into 100 mL of water and the pH was adjusted to 7-8 with saturated aqueous Na2CO3. The mixture was then extracted with EtOAc (100 mL x 6). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to afford UB2-012e (1100 mg, 90% yield) as a yellow solid. LCMS [M+1] + =257.

[0337] Step 4: UB2-012f: 2,4-dichloro-8-isopropylpyrazolo[1,5-a][1,3,5]triazine

[0338] UB2-012e (1100 mg, 4.28 mmol), triethylamine hydrochloride (1770 mg, 12.84 mmol, 3 eq), and POCl3 (50 mL) were added to a reaction flask and stirred at 110°C for 18 hours. The reaction solution was concentrated, dissolved in dichloromethane (20 mL), then added with ice water (50 mL) and extracted with dichloromethane (50 mL x 2). The organic phases were combined, concentrated, and purified by column chromatography (PE:EA = 20:1) to afford UB2-012f (510 mg, 52% yield) as a yellow oil. LCMS [M+1] + =231.

[0339] Step 5: UB2-012h: N-([1,1'-biphenyl]-4-ylmethyl)-2-chloro-8-isopropylpyrazolo[1,5-a][1,3,5]triazin-4-amine

[0340] To a reaction flask, UB2-012f (510 mg, 2.2 mmol, 1 eq), acetonitrile (30 mL), DIEA (851 mg, 6.6 mmol, 3 eq), and UB2-012g (403 mg, 2.2 mmol, 1 eq) were added. The reaction mixture was stirred at 70°C for 2 hours. The reaction mixture was concentrated, and the crude product was purified on a silica gel column (PE / EtOAc = 10 / 1 to 2 / 1) to afford UB2-012h (760 mg, 91% yield) as a yellow solid. LCMS [M+1] + =378.1.

[0341] Step 6: UB2-012j: (S)-tert-Butyl 4-(4-(([1,1'-biphenyl]-4-ylmethyl)amino)-8-isopropylpyrazolo[1,5-a][1,3,5]triazin-2-yl)-3-(hydroxymethyl)piperazine-1-carboxylate

[0342] To a reaction flask were added UB2-012h (430 mg, 1.276 mmol), UB2-012i (828 mg, 3.826 mmol), RuPhos Pd G2 (198 mg, 0.2552 mmol), t-BuONa (245 mg, 2.552 mmol), RuPhos (298 mg, 0.638 mmol), and 1,4-dioxane (20 mL). The reaction mixture was purged with argon three times, then heated to 100°C and stirred overnight. After completion of the reaction, the reaction mixture was dried and purified on a silica gel column (DCM / DCM in 10% MeOH) = 0-15% to afford UB2-012j (100 mg, 14% yield) as a yellow solid. LCMS [M+1] + =558.3.

[0343] Step 7: UB2-012: (S)-(1-(4-(([1,1'-biphenyl]-4-ylmethyl)amino)-8-isopropylpyrazolo[1,5-a][1,3,5]triazin-2-yl)piperazin-2-yl

[0344] UB2-012j (100 mg, 0.18 mmol) was dissolved in DCM (5 mL) and HCl / dioxane (1 mL, 4 M) was added. The reaction mixture was stirred at room temperature overnight. The reaction mixture was purified by Prep-HPLC (FA) to obtain UB2-012 (21.5 mg, 26% yield) as a white solid. LCMS [M+1] + =458.3.

[0345] 1H NMR(400MHz,DMSO-d6)δ8.93(t,J=6.3Hz,1H),7.74(s,1H),7.62(dd,J=11.8,4.7Hz,4H), 7.53-7.41(m,4H),7.34(t,J=7.3Hz,1H),4.63(d,J=6.1Hz,3H),4.45(d,J=13.0Hz,1H),3 .88-3.74(m,1H),3.46(dd,J=10.1,5.1Hz,1H),3.24(d,J=12.4Hz,1H),3.04-2.85(m,3H) ,2.71(dd,J=12.3,3.8Hz,1H),2.59(dd,J=16.6,8.7Hz,1H),1.22(dd,J=6.9,1.4Hz,6H).

[0346] Example 9: Synthesis of Compound UB2-013

[0347] Step 1: UB2-013b

[0348] Compound UB2-013a (800 mg, 4.26 mmol) was added to a round-bottom flask and dissolved in dry DMF (40 mL). Potassium carbonate (2.94 g, 21.3 mmol) and 2-bromopropane (1.57 g, 12.77 mol) were then added. The reaction mixture was allowed to react at 80°C for 3 hours, extracted with ethyl acetate, and the organic phase was washed three times with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (petroleum ether / ethyl acetate = 5 / 2) to obtain compound UB2-013b (659 mg, 67% yield) as a white solid. LCMS [M+1] + =230. NOE confirmed at the same time.

[0349] Step 2: UB2-013d

[0350] Compound UB2-013b (389 mg, 1.69 mmol), UB2-013c (1.70 g, 9.30 mol), and triethylamine (4 mL) were dissolved in n-butanol (8 mL) and microwave-treated at 180°C for 20 hours. The reaction mixture was cooled to room temperature, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (petroleum ether / ethyl acetate = 5 / 2). The mixture was then dried under vacuum to yield the target compound (UB2-013d, 480 mg, 75% yield) as a yellow solid. LCMS [M+1] + =377.2.

[0351] Step 3: UB2-013e

[0352] Compound UB-211202d (360 mg, 0.95 mmol) was dissolved in tetrahydrofuran (10 mL), and 4-dimethylaminopyridine (58 mg, 0.48 mmol), diisopropylethylamine (368 mg, 2.85 mmol), and di-tert-butyl dicarbonate (622 mg, 2.85 mol) were added sequentially. The reaction mixture was allowed to react at 50°C for 3 hours. After completion of the reaction, the reaction mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (petroleum ether / ethyl acetate = 2 / 1) to obtain compound UB2-013e (358 mg, 79% yield) as a white solid. LCMS [M+1] + =477.2.

[0353] Step 4: UB2-013g

[0354] Compound UB2-013e (250 mg, 0.52 mmol), UB2-013f (566 mg, 2.62 mol), tris(dibenzylideneacetone)dipalladium (238 mg, 0.26 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (248 mg, 0.52 mmol), cesium carbonate (508 mg, 1.56 mmol), and 1,4-dioxane (10 mL) were added to a 100 ml round-bottom flask. The atmosphere was replaced with argon three times and stirred at 100°C for 19 hours. After completion of the reaction, the reaction mixture was concentrated to dryness under reduced pressure, purified by column chromatography (petroleum ether / ethyl acetate = 1 / 2), and dried in vacuo to yield the target compound (UB2-013g, 130 mg, 38% yield) as a yellow solid. LCMS [M+1] + =657.4.

[0355] Step 5: UB2-013

[0356] Compound UB-211202g (130 mg, 0.2 mmol) was dissolved in dichloromethane (4 mL) and a dioxane hydrochloride solution (2 mL, 4 M) was added at room temperature. The reaction mixture was allowed to react at 25°C for 2 hours. After completion of the reaction, the reaction mixture was concentrated to dryness under reduced pressure and purified by reverse phase preparative purification to obtain compound (UB2-013, 30 mg, 33% yield) as a white solid. LCMS [M+1] + =457.3

[0357] 1H NMR (400MHz, DMSO-d6) δ7.89 (s, 1H), 7.63-7.59 (m, 2H), 7.56 (d, J = 8.3Hz, 2H), 7.47-7.40 (m, 4H), 7.33 (t,J=7.2Hz,1H),7.11(t,J=6.0Hz,1H),5.90(s,1H),4.63(d,J=6.0Hz,2H),4.49(dt,J=13.2,6.8Hz,1H ),4.26(s,1H),3.84(d,J=12.4Hz,1H),3.78-3.69(m,1H),3.35(d,J=6.0Hz,1H),3.26(d,J=12.0Hz,1H) ,3.01(d,J=12.0Hz,1H),2.92(dd,J=13.6,10.8Hz,1H),2.80-2.64(m,2H),1.45(dd,J=6.8,1.6Hz,6H).

[0358] Example 10: General method for synthesizing compound UB2-015

[0359] Step 1: UB2-015c: N-([1,1'-biphenyl]-4-ylmethyl)-5-chloro-3-isopropylpyrazolo[1,5-a]pyrimidin-7-amine

[0360] UB2-015a (500 mg, 2.05 mmol), ethanol (50 mL), DIEA (1300 mg, 10 mmol), and UB2-015b (720 mg, 4.0 mmol) were added to the reaction flask. The reaction mixture was stirred at 90°C for 2 hours. The reaction mixture was concentrated, and the crude product was purified on a silica gel column (PE / EtOAc = 10 / 1 to 2 / 1) to obtain UB2-015c (711 mg, 90% yield) as a yellow solid. LCMS [M+1] + =377.1.

[0361] Step 2: UB2-015d: tert-Butyl ([1,1'-biphenyl]-4-ylmethyl)(5-chloro-3-isopropylpyrazolo[1,5-a]pyrimidin-7-yl)carbamate

[0362] UB2-015c (200 mg, 0.532 mmol), DCM (5 mL), (Boc)2O (128 mg, 0.585 mmol), DIEA (343 mg, 2.66 mmol), and DMAP (32 mg, 0.266 mmol) were added to the reaction flask. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated, and the crude product was purified on a silica gel column (PE / EtOAc = 0-10%) to give UB2-015d (280 mg) as a white oil. LCMS [M+1] + =477.2.

[0363] Step 3: UB2-015f: (S)-tert-Butyl 4-(7-(([1,1'-biphenyl]-4-ylmethyl)(tert-butoxycarbonyl)amino)-3-isopropylpyrazolo[1,5-a]pyrimidin-5-yl)-3-(hydroxymethyl)piperazine-1-carboxylate

[0364] UB2-015d (230 mg, 0.483 mmol), UB2-015e (314 mg, 1.45 mmol), Pd2(dba)3 (44 mg, 0.0483 mmol), Cs2CO3 (315 mg, 0.966 mmol), rac-BINAP (150 mg, 0.24 mmol), and 1,4-dioxane (15 mL) were added to the reaction flask. The reaction mixture was purged with argon three times, then heated to 100°C and stirred overnight. After the reaction was complete, the reaction mixture was dried and purified on a silica gel column (DCM / (DCM / MeOH=10 / 1)=0-15%) to afford UB2-015f (80 mg, 30% yield) as a yellow solid. LCMS [M+1] + =657.4.

[0365] Step 4: UB2-015: (S)-(1-(7-(([1,1'-biphenyl]-4-ylmethyl)amino)-3-isopropylpyrazolo[1,5-a]pyrimidin-5-yl)piperazin-2-yl)methanol

[0366] UB2-015f (85 mg, 0.15 mmol) was dissolved in DCM (100 mL) and HCl / dioxane (3 mL, 4 M) was added. The reaction mixture was stirred at room temperature overnight. The reaction mixture was purified by Prep-HPLC (FA) to give UB2-015 (30.5 mg, 50% yield) as a white solid. LCMS [M+1] + =457.3.

[0367] 1H NMR (400MHz, DMSO-d6) δ8.03(t,J=6.6Hz,1H),7.70(s,1H),7.66-7.59(m,4H),7.50(d,J=8.3Hz,2H),7. 45(dd,J=10.4,4.8Hz,2H),7.35(t,J=7.3Hz,1H),5.59(s,1H),4.60(d,J=6.6Hz,2H),4.42-4.21(m,2H), 3.75(dd,J=10.5,8.2Hz,1H),3.49(dd,J=10.5,5.0Hz,1H),3.38(d,J=12.6Hz,1H),3.16(d,J=11.9Hz,1 H),3.06(dd,J=20.7,8.3Hz,1H),3.02-2.90(m,2H),2.81(td,J=12.1,3.5Hz,1H),1.25(d,J=6.9Hz,6H).

[0368] Example 9: General method for synthesizing compound UB2-018

[0369] Step 1: UB2-018b: 2-Bromo-3-methylbutanal

[0370] UB2-018a (5.0 g, 58.15 mmol) was dissolved in 50 mL of dichloromethane, and L-proline (0.67 g, 5.82 mmol) was added. N-bromosuccinimide (12.4 g, 69.65 mmol) was added to the solution under ice-cooling, and stirred at room temperature for 2 hours. After filtration, the reaction mixture was concentrated and purified by flash chromatography (eluent: PE / EtOAc = 0-50%) to obtain a yellow solid compound (UB2-018b, 5.0 g, 52.4% yield). LCMS [M+1] + =165,167.

[0371] Step 2: UB2-018d: 8-bromo-6-chloro-3-isopropylimidazo[1,2-b]pyridazine

[0372] UB2-018b (5.0 g, 30.5 mmol) was dissolved in 50 ml of ethanol, and UB2-018c (1.25 g, 6.1 mmol) was added. The mixture was stirred at 80°C for 48 hours. Water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was collected, dried, and the solvent removed. The concentrate was purified by flash chromatography (eluent: PE / EtOAc = 0-50%) to obtain UB2-018d (1.5 g, 18% yield) as a white solid. LCMS [M+1] + =274,276.

[0373] Step 3: UB2-018f: N-([1,1'-biphenyl]-4-ylmethyl)-6-chloro-3-isopropylimidazo[1,2-b]pyridazin-8-amine

[0374] UB2-018d (1.5 g, 5.5 mmol) was dissolved in 40 mL of ethanol, and UB2-018e (1.0 g, 5.5 mmol) and N,N-diisopropylethylamine (2.2 g, 16.5 mmol) were added. The mixture was stirred at 80°C overnight. After removing the solvent, the concentrate was purified by flash chromatography (eluent: DCM / MeOH = 10:1) to obtain UB2-018f (1.1 g, 51.8% yield) as a white solid. LCMS [M+1] + =377.2.

[0375] Step 4: UB2-018g: tert-Butyl ([1,1'-biphenyl]-4-ylmethyl)(6-chloro-3-isopropylimidazo[1,2-b]pyridazin-8-yl)carbamate

[0376] UB2-018f (350 mg, 0.93 mmol) was dissolved in 8 mL of tetrahydrofuran, and 4-dimethylaminopyridine (11 mg, 0.09 mmol), N,N-diisopropylethylamine (1.2 g, 9.3 mmol), and di-tert-butyl dicarbonate (2 g, 9.3 mmol) were added. The mixture was stirred at 50°C overnight. After removing the solvent, the concentrate was purified by flash chromatography (eluent: DCM / MeOH = 10:1) to obtain UB2-018g (400 mg, 90.2% yield) as a white solid. LCMS [M+1] + =477.2.

[0377] Step 5: UB2-018i: (S)-tert-Butyl 4-(8-(([1,1'-biphenyl]-4-ylmethyl)(tert-butoxycarbonyl)amino)-3-isopropylimidazo[1,2-b]pyridazin-6-yl)-3-(hydroxymethyl)piperazine-1-carboxylate

[0378] UB2-018g (150 mg, 0.31 mmol) was dissolved in 20 mL of dioxane solution, and UB2-018h (204 mg, 0.95 mmol), Pd2(dba)3 (29 mg, 0.03 mmol), rac-BINAP (40 mg, 0.065 mmol), and cesium carbonate (320 mg, 1.0 mmol) were added. The mixture was stirred at 105°C overnight under an argon atmosphere. After removing the solvent, the concentrate was purified by flash chromatography (eluent: DCM / MeOH = 10:1) to obtain UB2-018i (137 mg, 66.5% yield) as a yellow solid. LCMS [M+1] + =657.4.

[0379] Step 6: UB2-018: (S)-(1-(8-(([1,1'-biphenyl]-4-ylmethyl)amino)-3-isopropylimidazo[1,2-b]pyridazin-6-yl)piperazin-2-yl)methanol

[0380] UB2-018i (137 mg, 0.19 mmol) was dissolved in 10 mL of dichloromethane, and 5 mL of 4 M hydrochloric acid in dioxane was added. The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the crude product was purified by preparative liquid chromatography (mobile phase: formic acid / water / acetonitrile) to obtain UB2-018 (32 mg, yield: 34%) as a white solid. LCMS [M+1] + =457.3. 1 H NMR (400MHz, DMSO-d6) δ7.65-7.55(m,5H),7.51-7.41(m,4H),7.34(t,J=7.3Hz,1H),7.08(s,1H ),5.80(s,1H),4.56(d,J=6.4Hz,2H),3.99-3.97(m,1H),3.85-3.82(m,1H),3.79-3.76(m,1H), 3.41(dd,J=10.3,4.6Hz,1H),3.25(t,J=9.6Hz,1H),3.18(dd,J=13.7,6.9Hz,1H),3.01(dd,J=2 1.5,11.7Hz,2H),2.83(dd,J=12.4,3.9Hz,1H),2.77-2.67(m,1H),1.29(dd,J=6.9,4.2Hz,6H).

[0381] Example 10

[0382] Other compounds listed in Table A can be synthesized by referring to the methods of Examples 1-9 by replacing the corresponding raw materials.

[0383] Table A

[0384] B. Test Examples

[0385] Test Example 1: Cell proliferation experiment

[0386] Reagents: RPMI-1640 medium, McCoy's 5A medium, IMDM medium, MEM medium, L-15 medium, fetal bovine serum, blue-chain double antibody, trypsin, 2-mercaptoethanol, NEAA, pyruvate, etc.

[0387] The test cells were routinely cultured and passaged for at least 2 generations before plating.

[0388] 1000 HepG2 cells in 25uL were seeded into a 384-well plate with black walls and transparent bottom, and 25uL of compounds at different concentrations were added. The cells were cultured overnight at 37°C and 5% CO2.

[0389] 1500 H82 and H69 cells were seeded in 25uL of 384-well plates with black walls and transparent bottoms, and 25uL of compounds at different concentrations were added to culture the cells at 37°C and 5% CO2 for 72 hours. The 384-well plates were equilibrated at room temperature, and 25uL of Cell Reagent, shake on a horizontal shaker for 2 minutes to lyse the cells, incubate at room temperature for 10 minutes to stabilize the luminescent signal, and then use Envision to detect the chemiluminescent signal.

[0390] All cells were subjected to IC of the corresponding test samples. 50 Determination.

[0391] The fluorescence intensity of each well was detected using the Alarm blue method, and the IC 50 .

[0392] IC 50 Calculated by the following formula: Y=Max+(Min-Max) / [1+(X / IC 50 )×Slope]

[0393] Where Min, Max, and Slope represent the minimum value, maximum value, and slope, respectively.

[0394] The results are shown in Table 1. The compounds of the present invention were subjected to cell inhibition experiments on tumor cell lines. The results showed that most of the compounds exhibited strong cell killing effects in tumor cells and have the potential to be developed as anti-tumor drugs.

[0395] Table 1

[0396] In the table, A≤50nM, 50nM <B≤100nM,100nM<C≤300nM,D> 300nM.

[0397] Test Example 2: Immunohistochemistry (IHC)

[0398] Add appropriate amount of compound to the cultured cells and incubate in a 37°C tissue culture incubator for 2 hours.

[0399] The cultured cells were washed twice with PBS and then fixed with 100 μL of 10% neutral buffered formalin for 20 minutes.

[0400] After washing three times with PBS, immunohistochemical staining was performed using CCNK antibody and a universal two-step detection kit (mouse / rabbit enhanced polymer detection system, PV-9000, Zsbio Inc, Beijing).

[0401] Specifically, 100 μL of the endogenous peroxidase blocker in the kit was added and incubated at room temperature for 10 minutes; and then rinsed with PBS buffer for 3 minutes × 3 times.

[0402] Add 100 μL of CCNK antibody diluted 1:5000 and incubate at 37°C for 60 minutes; then rinse with PBS buffer for 3 minutes × 3 times.

[0403] Add 100 μL of reaction enhancement solution and incubate at 37°C for 20 minutes; rinse with PBS buffer for 3 minutes × 3 times.

[0404] Add 100 μL of enhanced enzyme-labeled goat anti-mouse / rabbit IgG polymer and incubate at 37°C for 20 minutes; rinse with PBS buffer for 3 minutes × 3 times.

[0405] Place in 75% ethanol, soak for 3 minutes × 2 times; place in 95% ethanol, soak for 3 minutes × 2 times; after removing excess liquid, place in anhydrous ethanol, soak for 3 minutes × 3 times; after removing excess liquid, air dry, seal the slide and observe under a microscope and take pictures.

[0406] Test Example 3: Western blotting

[0407] A transparent 12-well plate was coated with 500 μL of 0.01 mg / mL poly-D-lysine hydrobromide and incubated at 37°C for 1 hour. The coating solution was removed and the plate was washed twice with 1 mL of PBS. 400,000 HEK293 cells were seeded into the 12-well plate. The cells were treated with the compound for 24 hours. The medium was removed, the plate was washed with PBS, and the cells were lysed by adding RIPA buffer. An appropriate volume of the cell lysate was added to the loading buffer and slowly added to the corresponding wells of the plate for SDS-PAGE (4%-12%). After the run, the plate was transferred to a PVDF membrane and blocked with 5% skim milk powder for 1 hour at room temperature. The membrane was then incubated with an anti-cyclin K primary antibody diluted in 5% skim milk powder and shaken overnight at 4°C. After the primary antibody incubation, the membrane was washed three times with TBST on a shaker. An anti-rabbit HRP secondary antibody corresponding to the primary antibody was added in 5% skim milk powder and shaken for 1 hour at room temperature. After secondary antibody incubation, wash the membrane three times with TBST on a shaker. Place the PVDF membrane flat in a dark box, evenly soak the bands with ECL developer, and photograph using a ChemDoc XRS+ gel imager. Quantitatively analyze protein band intensities using ImageJ software.

[0408] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that: The compound is as shown in formula (I); in, R 1 Each independently is H or C 1-4 alkyl; Subscript n1 is 1, 2, or 3; Cyclic Ar 1 Select from the following group: C 6-10 Aromatic ring, 5- to 10-membered heteroaromatic ring, 5- to 10-membered bridged ring; Cyclic Cr 1 Selected from the following group: H, C 3-10 Carbocyclic group, 3 to 10 membered heterocyclic group, C 6-10 aryl, and 5- to 10-membered heteroaryl; R a and R b Each independently selected from the group consisting of: H, R e or R; or R a and R b With cyclic Ar 1 and cyclic Cr 1 Jointly formed in, X 7 Each independently selected from the group consisting of: -O-, -S-, -N(R c )-、-C(R c )2-、-C(R c )2-C(R c )2-; Subscripts n5 and n6 are each independently 0, 1, 2 or 3; R e Each independently selected from the group consisting of: hydroxyl, C 1-6 Alkyl, -OC 1-6 Alkyl, -OC 1-6 Alkylene-R f ; Among them, R f Selected from the group consisting of: -CN, -OH, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2; Subscripts n3 and n4 are each independently 0, 1, 2, 3 or 4; R 2 Selected from the group consisting of H, CN, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-8 cycloalkyl, optionally substituted 3- to 8-membered heterocyclyl; X 1 and X 2 Each independently is N or C(R c ); X 3 、X 4 and X 5 are each independently N or C; X 6 N, N(R c )、C(R c ), O or S; M 1 Select from the following groups: None, X 8 、(M 4 ) s ;in, X 8 N(R c ) or C(R c )2; M 4 Each independently selected from the group consisting of O, S, C(O)O, C(O), N(R c ) and C 1-4 alkylene; s is 1, 2, or 3; M 2 is none or a ring as shown in formula A; In formula A, X 9 Representative and M 1 Connected position, and X 9 N or C(R m ); X 10 Representative and M 3 Connected position, and X 10 Selected from the group consisting of O, S, N, or C(R m ); Each X 11 are independently -C(R m )2-or-N(R m )-; Subscripts m1 and m2 are each independently 0, 1, 2 or 3, and m1+m2≥2; R m Each independently is R c or R 5 ; or, two R m Together form a single bond, optionally substituted C 1-4 alkylene or optionally substituted 1- to 4-membered heteroalkylene; R 5 Each independently selected from the group consisting of: hydroxy, optionally substituted C 1-6 Hydroxyalkyl, optionally substituted -C(O)-NH2, optionally substituted -C(O)-NH(C 1-6- alkyl), optionally substituted -C(O)-N(C 1-6- alkyl)2, optionally substituted C 1-6 Alkyl, optionally substituted -C(O)-C 1-6- Alkyl and optionally substituted C 1-6 haloalkyl; or, two R on the same carbon atom 5 Together they form an oxo group (=O) or a thio group (=S); M 3 Selected from the group consisting of: H, None, R 3 、-NH-R 4 ; Among them, R 3 Selected from the group consisting of hydroxy, optionally substituted C 1-6- Hydroxyalkyl, optionally substituted -C(O)-C 1-6- alkyl, optionally substituted-C(O)-NH2, optionally substituted-C(O)-NH(C 1-6- alkyl), optionally substituted -C(O)-N(C 1-6- alkyl)2, optionally substituted C 1-6- alkyl halide; R 4 Selected from the group consisting of: H, optionally substituted C 1-6 Alkyl, optionally substituted C 1-6- Hydroxyalkyl, optionally substituted C 1-6- alkyl halide; R c Each independently is H or C 1-4 alkyl; Unless otherwise defined, the optional substitution refers to unsubstituted or one or more (such as 1, 2, 3 or 4) hydrogen in the group is replaced by a substituent R, and R is selected from the following groups: D, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, -CN, -OR', -NO2, -NR'R", -SR', -OC(O)R', -C(O)R', -CO2R', -CONR', -OC(O)NR'R", -NR"C(O)R', -NR"-C(O)NR'R", -NR"C(O)2R', -S(O)R', -S(O)2R', -S(O)2NR'R", -NR"S(O)2R', C(O)2R', optionally substituted with one or more R' 3-10 Cycloalkyl, 4 to 10 membered heterocycloalkyl optionally substituted by one or more R'", C 6-10 aryl, 5- to 10-membered heteroaryl optionally substituted by one or more R'", -C 1-4 Alkylene-C 3-10 Cycloalkyl, optionally substituted with one or more R'"-C 1-4 Alkylene-4 to 10 membered heterocycloalkyl, optionally substituted with one or more R'"-C 1-4 Alkylene-C 6-10 Aryl, optionally substituted with one or more R'" -C 1-4 Alkylene-5 to 10 membered heteroaryl; Each R' is independently H, D, or a group selected from the group consisting of: C 1-6 Alkyl, C 3-10 Cycloalkyl, 4- to 10-membered heterocycloalkyl, C 6-10 Aryl, 5- to 10-membered heteroaryl, -C 1-4 Alkylene-C 3-10 Cycloalkyl, -C 1-4 Alkylene-4 to 10 membered heterocycloalkyl, -C 1-4 Alkylene-C 6-10 Aryl-C 1-4 Alkylene-5 to 10 membered heteroaryl; Each R" is selected from the group consisting of H, D, C 1-4 Alkyl, C 1-4 Haloalkyl, and C 3-4 Cycloalkyl; Each R"' is independently selected from the group consisting of D, halogen, hydroxy, nitro, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein The compound is as described in Formula II Among them, X 1 、X 2 、X 3 、X 4 、X 5 、X 6 、M 1 、M 2 、M 3 、R 2 As defined in Formula I.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein The compound is shown in formula II-1 or II-2 in, In formula II-1, X 9 N, X 10 N, X 11 One of them is -CH(R 5 )-, -C(O)- or -C(S)-, the rest X 11 is -CH2-; and R 5 is optionally substituted C 1-6 hydroxyalkyl or optionally substituted -C(O)-NH2; In formula II-2, X 9 CH, X 10 CH, X 11 All -CH2-; and R 3 Selected from the group consisting of hydroxy, optionally substituted C 1-6- Hydroxyalkyl.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein The compound is selected from Table A, Table B1 or Table B2.

5. A pharmaceutical composition comprising: (i) the compound according to claim 1 or a pharmaceutically acceptable salt thereof; as well as (ii) a pharmaceutically acceptable carrier.

6. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of (a) a drug for treating cancer and / or (b) a cell cycle protein degrader.

7. A conjugate or prodrug, characterized in that The conjugate or prodrug is a compound comprising a molecular glue degrading agent portion or a pharmaceutically acceptable salt thereof, wherein the molecular glue degrading agent portion is derived from the compound according to claim 1.

8. The conjugate or prodrug according to claim 7, wherein (i) The conjugate is a compound represented by Formula III or a pharmaceutically acceptable salt thereof; M D -M CL -M CP (III) Among them, M D It is the molecular glue degradation agent part; M CL No or connector part; M CP is a portion derived from a polypeptide element or a targeting ligand; (ii) the prodrug is a compound represented by Formula IV or a pharmaceutically acceptable salt thereof; M D -M PL -M LG (4) Among them, M D It is the molecular glue degradation agent part; M PL Is none or a connecting group; M LG For the leaving part.

9. A pharmaceutical composition comprising: (i) the conjugate or prodrug according to claim 7; as well as (ii) a pharmaceutically acceptable carrier.

10. Use of the conjugate or prodrug according to claim 7 in the preparation of a medicament for treating cancer.

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