Combination therapy methods for treating TP53-Y220C variant leukemia and TP53 wild-type leukemia.

Abstract

This disclosure provides methods for treating TP53-Y220C variant leukemias and TP53 wild-type leukemias, such as acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). In some embodiments, the methods disclosed herein include administering an indole derivative to a target in combination with one or more of the following: an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor. TIFF2026522611000101.tif178128

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims the benefits and priority of U.S. Provisional Patent Application No. 63 / 508,949, filed June 19, 2023, which is incorporated herein by reference in its entirety for all purposes.

[0002] Technical field This technology, as a whole, relates to a method for treating TP53-Y220C variant leukemia and TP53 wild-type leukemia, such as acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). [Background technology]

[0003] background The following background explanation of this technology is provided solely to aid in understanding this technology and does not constitute an endorsement of prior art.

[0004] The tumor suppressor network is an elaborate network capable of blocking cells that harbor activated oncogenes, damaged genomes, or other cancer-promoting mutations derived from replication. A central component of the tumor suppressor network is p53, one of the most potent tumor suppressors in cells. Both the wild-type and mutant conformations of p53 are involved in cancer progression. TP53-Y220C is a recurrent hotspot TP53 mutation predominantly found in solid tumors and hematological malignancies in AML and MDS. It occurs frequently as a subclone among TP53-WT cancer cells. On the other hand, the p53 reactivator PC14586 primarily exerts a cell division inhibitory effect, and therefore its apoptotic activity in TP53-Y220C leukemia cells is limited.

[0005] Consequently, there is an urgent need to identify clinically feasible treatment strategies for acute myeloid leukemia (AML) and MDS patients with the TP53-Y220C mutation.

Summary of the Invention

[0006] Summary of the Technology In one aspect, the present disclosure provides a method for treating wild-type p53 leukemia or TP53-Y220C leukemia in a patient in need thereof, the method comprising administering to the patient an effective amount of a TP53 reactivation indole derivative and an effective amount of an MDM2 inhibitor, a BCL-2 inhibitor, and / or an XPO-1 inhibitor, wherein the TP53 reactivation indole derivative has the formula (I): TIFF2026522611000002.tif29128 or a pharmaceutically acceptable salt thereof, wherein, each TIFF2026522611000003.tif2128 is independently a single bond or a double bond; X 6 is a carbon atom linked to CR 5 CR 5 R 6 N, NR 5 O, S, C=O, C=S, or Q 1 ; X 2 is a carbon atom linked to CR 7 CR 7 R 8 N, NR 7 O, S, C=O, C=S, or Q 1 ; X 3 is a carbon atom linked to CR 9 CR 9 R 10 N, NR 9 O, S, C=O, C=S, or Q 1 ;<0​​​​​​​​​11 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 5 CR 13 , N, or NR 13 and; X 1 , X 2 , X 3 , and X 4 At least one of them is Q 1 It is a carbon atom linked to; Q 1 C=O, C=S, C=CR 14 R 15 , C=NR 14 , alkylene, alkenylene, or alkynylene (each of these independently substituted or unsubstituted), bond; m is 1, 2, 3, or 4; Y is either N or O; R 1 is -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 , -R 16 Ure 17 , -R 16 Ure 17 R 18 , -SR 16 , -R 16 SR 17 , -R 16 SR 17 R 18 , -NR 16 R 17 , -R 16 NR 17 , -R 16 NR 17 R 18 , -NR 16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18、 alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which may be independently substituted or unsubstituted), or hydrogen; each R 3 and R 4 is independently -C(O)R 19 、 -C(O)OR 19 、 -C(O)NR 19 R 20 、 -SOR 19 、 -SO2R 19 、 alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which may be independently substituted or unsubstituted), or hydrogen; each R 2 、 R 5 、 R 6 、 R 7 、 R 8 、 R 9 、 R 10 、 R 11 、 R 12 、 R 13 、 R 14 、 R 15 、 R 16 、 R 17 、 and R 18 is independently -C(O)R 21 、 -C(O)OR 21 、 -C(O)NR 21 R 22 、 -OR 21 、 -SR 21 、 -NR 21 R 22 、 -NR 21 C(O)R 22 、 -OC(O)R 21 、 halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which may be independently substituted or unsubstituted), or hydrogen; each R 19 and R 20 is C(O)R 23 、 -C(O)OR 23 、 -C(O)NR 23 R 24, -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R 21 and R 22 is independently an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R 23 and R 24 These are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen.

[0007] In one aspect, the present disclosure provides a method for extending the survival time of patients with wild-type p53 leukemia or TP53-Y220C leukemia, comprising the step of administering to the patient an effective amount of a TP53 reactivating indole derivative and an effective amount of an MDM2 inhibitor, a BCL-2 inhibitor, and / or an XPO-1 inhibitor, wherein the TP53 reactivating indole derivative is of formula (I): Having the formula TIFF2026522611000004.tif29128, or a pharmaceutically acceptable salt thereof, During the ceremony, each TIFF2026522611000005.tif2128 is independently either a single bond or a double bond; X 1 CR 5 , CR 5 R 6 , N, NR 5 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X2 CR 7 , CR 7 R 8 , N, NR 7 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 3 CR 9 , CR 9 R 10 , N, NR 9 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 4 CR 11 , CR 11 R 12 , N, NR 11 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 5 CR 13 , N, or NR 13 and; X 1 , X 2 , X 3 , and X 4 At least one of them is Q 1 It is a carbon atom linked to; Q 1 C=O, C=S, C=CR 14 R 15 , C=NR 14 , alkylene, alkenylene, or alkynylene (each of these independently substituted or unsubstituted), bond; m is 1, 2, 3, or 4; Y is either N or O; R 1 is -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 , -R 16 Ure 17 , -R 16 Ure 17 R 18 , -SR 16 , -R16 SR 17 , -R 16 SR 17 R 18 , -NR 16 R 17 , -R 16 NR 17 , -R 16 NR 17 R 18 , -NR 16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 -C(O)R 19 , -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 , -SO2R 19 , alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently unsubstituted or substituted), or hydrogen; Each R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -NR21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 , halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently unsubstituted or substituted), or hydrogen; Each R 19 and R 20 C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R 21 and R 22 is independently an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R 23 and R 24 These are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen.

[0008] In one aspect, the present disclosure provides a method for selecting leukemia patients for treatment with TP53 reactivated indole derivatives and additional therapeutic substances, comprising the steps of detecting the mRNA or polypeptide expression of wild-type TP53 or TP53-Y220C in a biological sample obtained from a leukemia patient, and administering to the patient an effective amount of TP53 reactivated indole derivatives and an effective amount of an MDM2 inhibitor, a BCL-2 inhibitor, and / or an XPO-1 inhibitor, wherein the TP53 reactivated indole derivatives are of formula (I): Having the formula TIFF2026522611000006.tif29128, or a pharmaceutically acceptable salt thereof, During the ceremony, each TIFF2026522611000007.tif2128 is independently either a single bond or a double bond; X 1 CR 5 , CR 5 R 6 , N, NR 5 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 2 CR 7 , CR 7 R 8 , N, NR 7 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 3 CR 9 , CR 9 R 10 , N, NR 9 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 4 CR 11 , CR 11 R 12 , N, NR 11 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 5 CR 13 , N, or NR13 and; X 1 , X 2 , X 3 , and X 4 At least one of them is Q 1 It is a carbon atom linked to; Q 1 C=O, C=S, C=CR 14 R 15 , C=NR 14 , alkylene, alkenylene, or alkynylene (each of these independently substituted or unsubstituted), bond; m is 1, 2, 3, or 4; Y is either N or O; R 1 is -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 , -R 16 Ure 17 , -R 16 Ure 17 R 18 , -SR 16 , -R 16 SR 17 , -R 16 SR 17 R 18 , -NR 16 R 17 , -R 16 NR 17 , -R 16 NR 17 R 18 , -NR 16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 -C(O)R 19, -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 , -SO2R 19 , alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently unsubstituted or substituted), or hydrogen; Each R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 , halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently unsubstituted or substituted), or hydrogen; Each R 19 and R 20 C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R 21 and R 22 is independently an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R 23 and R 24 These are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen.

[0009] The process of administering to a leukemia patient may include administering an effective amount of a TP53 reactivated indole derivative and an effective amount of an MDM2 inhibitor. The process of administering to a leukemia patient may include administering an effective amount of a TP53 reactivated indole derivative and an effective amount of a BCL-2 inhibitor. The process of administering to a leukemia patient may include administering an effective amount of a TP53 reactivated indole derivative and an effective amount of an XPO-1 inhibitor. The process of administering to a leukemia patient may include administering an effective amount of a TP53 reactivated indole derivative, an effective amount of an MDM2 inhibitor, and an effective amount of a BCL-2 inhibitor. The process of administering to a leukemia patient may include administering an effective amount of a TP53 reactivated indole derivative, an effective amount of an MDM2 inhibitor, and an effective amount of an XPO-1 inhibitor. The process of administering to a leukemia patient may include administering an effective amount of a TP53 reactivated indole derivative, an effective amount of a BCL-2 inhibitor, and an effective amount of an XPO-1 inhibitor. The process of administering to a leukemia patient may include administering an effective amount of a TP53 reactivated indole derivative, an effective amount of an MDM2 inhibitor, an effective amount of a BCL-2 inhibitor, and an effective amount of an XPO-1 inhibitor.

[0010] Examples of MDM2 inhibitors include, but are not limited to, RG7112, RO5045337, Idasanutrin, Nutrin-3a, RG7388, AMG-232, KRT-232, APG-115, BI-907828, CGM097, Cilemadrin, HDM201, Mirademethan, BI907828, MEL23, MEL24, and DS-3032b, as well as pyrooxyindole MDM2 inhibitors tethered to lenalidomide (e.g., MI-1061), Nutrin derivatives tethered to lenalidomide analogs, YX-02-030 (aRG7112 derivative), and MDM2 degrading agents such as MS3227. Examples of BCL-2 inhibitors include, but are not limited to, venetoclax, ovatoclax, subatoclax, maritoclax, gossypol, apogossypol, TW-37, UMI-77, APG2575, and BDA-366. Examples of XPO-1 inhibitors include, but are not limited to, KPT330 (selinexol), XPOVIO, KPT8602 (ertanexol), KPT8602, KPT330, KPT335, verginexol, and KPT185.

[0011] Additionally or alternatively, in any embodiment of the methods disclosed herein, the MDM2 inhibitor, BCL-2 inhibitor, and / or XPO-1 inhibitor are administered sequentially, simultaneously, or separately with the TP53 reactivated indole derivative. In certain embodiments, the MDM2 inhibitor, BCL-2 inhibitor, XPO-1 inhibitor, and / or TP53 reactivated indole derivative are administered orally, intravenously, intramuscularly, intraperitoneally, or subcutaneously.

[0012] In any aspect of the methods disclosed herein, leukemia is acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS). Leukemia may be TP53-Y220C leukemia.

[0013] The TP53 reactivated indole derivative is given by formula (IA): It may have the formula TIFF2026522611000008.tif29128, or may be a pharmaceutically acceptable salt thereof. During the ceremony, X 1 CR 5 , NR 5 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 2 CR 7 CH, NR 7 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 3 CR 9 CH, NR 9 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 4 CR 11 CH, NR 11 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 5 CR 13 CH, or NR 13 and; X 1 , X 2 , X 3 , and X 4 At least one of them is Q 1 It is a carbon atom linked to; Q 1 It is a combination; m is 1, 2, 3, or 4; Y is either N or O; R 1 is -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 , -R 16 Ure 17 , -R 16 Ure 17 R 18 , -SR 16 , -R16 SR 17 , -R 16 SR 17 R 18 , -NR 16 R 17 , -R 16 NR 17 , -R 16 NR 17 R 18 , -NR 16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 -C(O)R 19 , -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 , -SO2R 19 , hydrogen, alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl, where each of alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl is independently unsubstituted, or substituted with one or two groups selected from alkyl or halogen, or R 3 and R 4 R 3 and R 4 It forms a ring together with the bonded Y atom, where the ring is either substituted or unsubstituted, or R 3 He is absent; Each R 2 , R 5 , R 7 , R 9 , R 11 , R 13 , R 16 , R 17, and R 18 -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 The elements are hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, and each alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl is independently unsubstituted or -C(O)R 25 , -C(O)OR 25 -C(O)NR 25 ,-CR 25 3, -OR 25 , -SR 25 , -NR 25 R 26 , -NR 25 C(O)R 26 -OC(O)R 25 It is substituted with one or two groups selected from alkyl, alkenyl, or alkynyl groups, R 25 and R 26 Each of these is independently an alkyl, hydrogen, or halogen; Each R 19 and R 20 C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R21 and R 22 is independently an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R 23 and R 24 These are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen.

[0014] The TP53 reactivated indole derivative is given by formula (IB): It may have the formula TIFF2026522611000009.tif38128, or may be a pharmaceutically acceptable salt thereof. During the ceremony, X 1 CR 5 , NR 5 , a carbon atom linked with O, S, C=O, C=S, or Y; X 2 CR 7 CH, NR 7 , a carbon atom linked with O, S, C=O, C=S, or Y; X 3 CR 9 CH, NR 9 , a carbon atom linked with O, S, C=O, C=S, or Y; X 4 CR 11 CH, NR 11 , a carbon atom linked with O, S, C=O, C=S, or Y; X 5 CR 13 CH, or NR 13 and; X 1 , X 2 , X 3 , and X 4 At least one of them is a carbon atom linked to Y; Y is either N or O; R1 is -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 , -R 16 Ure 17 , -R 16 Ure 17 R 18 , -SR 16 , -R 16 SR 17 , -R 16 SR 17 R 18 , -NR 16 R 17 , -R 16 NR 17 , -R 16 NR 17 R 18 , -NR 16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 -C(O)R 19 , -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 , -SO2R 19 , hydrogen, alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl, where each of alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl is independently unsubstituted, or substituted with one or two groups selected from alkyl or halogen, or R 3 and R 4 R 3 and R 4It forms a ring together with the bonded Y atom, where the ring is either substituted or unsubstituted, or R 3 He is absent; Each R 2 , R 5 , R 7 , R 9 , R 11 , R 13 , R 16 , R 17 , and R 18 -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 ,-CR 25 3, -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 , hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, and each of the alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl is independently unsubstituted or -C(O)R 25 , -C(O)OR 25 -C(O)NR 25 ,-CR 25 3, -OR 25 , -SR 25 , -NR 25 R 26 , -NR 25 C(O)R 26 -OC(O)R 25 It is substituted with one or two groups selected from alkyl, alkenyl, or alkynyl groups, R 25 and R 26 Each of these is independently an alkyl, hydrogen, or halogen; Each R 19 and R 20 C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R 21 and R 22 is independently an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R 23 and R 24 These are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen.

[0015] The TP53 reactivated indole derivative is given by formula: It may have TIFF2026522611000010.tif41128.

[0016] Additionally or alternatively, in any aspect of the methods disclosed herein, mRNA expression levels are detected by real-time quantitative PCR (qPCR), digital PCR (dPCR), reverse transcriptase-PCR (RT-PCR), Northern blotting, microarrays, dot or slot blotting, in-situ hybridization, or fluorescence in-situ hybridization (FISH). Additionally or alternatively, in certain aspects of the methods disclosed herein, polypeptide expression levels are detected by Western blotting, enzyme-linked immunosorbent assay (ELISA), dot blotting, immunohistochemistry, immunofluorescence, immunoprecipitation, immunoelectrophoresis, or mass spectrometry.

[0017] In any aspect of the methods disclosed herein, the patient is unresponsive to at least one prior cancer treatment, such as chemotherapy or immunotherapy. In some aspects, the chemotherapy is trioxide, azacitidine, cerubidine, cyclophosphamide, cytarabine, daunorubicin hydrochloride, daurismo, dexamethasone, doxorubicin hydrochloride, enasidenib mesylate, gemtuzumab ozogamicin, gilteritinib fumarate, grasdecib maleate, idamycin PFS, idarubicin hydrochloride, idhifa, ivosidenib, midostaurine, mitoxantrone hydrochloride, mylotarg, o The medication may include one or more of the following: rutasidenib, onureg, pemazyre, pemigatinib, prednisone, rezlidhia, rituxan, rituximab, rubidomycin, rydapt, tabloid, thioguanine, tibsovo, tisagenlecleucel, trisenox, venclexta, venetoclas, vinicristine sulfate, vyxeos, or xospata. Patients may be children or adults.

[0018] Also disclosed herein are kits comprising (a) a TP53 reactivating indole derivative (e.g., PC14586), (b) at least one of an MDM2 inhibitor, a BCL-2 inhibitor, and / or an XPO-1 inhibitor, and (c) instructions for treating wild-type p53 leukemia and / or TP53-Y220C leukemia. [Brief explanation of the drawing]

[0019] [Figure 1]PC14586 converts mutant p53 Y220C to the wild-type p53 conformation (upper panel), activates p53 transcriptional activity, and greatly induces p21 (lower panel). Figure 1 shows Western blots demonstrating the activity of PC14586 against mutant and wild-type p53 (upper panel) and Western blots demonstrating the transcriptional activity of p53 after treatment with PC14586 (lower panel). [Figure 2] Figures 2A-2B: PC14586 primarily inhibits cell proliferation in TP53 Y220C AML cells, but does not inhibit cell proliferation in TP53 WT AML cells, TP53 KO AML cells, or TP53 R175H mutant AML cells. Figure 2A: Dose-response survival curves for a group of AML cell lines containing TP53-WT, TP53-KO, TP53-R175H, or TP53-Y220C treated with PC14586 for 120 hours. Figure 2B: 7-aminoactinomycin (7AAD) / annexin V (AnnV) curves for a group of AML cell lines containing TP53-WT, TP53-KO, TP53-R175H, or TP53-Y220C treated with PC14586 for 120 hours. [Figure 3A] Figures 3A-3H: MDM2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 3A: 7AAD / AnnV curves of AML cells with TP53-WT treated for 72 hours with an MDM2 inhibitor, PC14586, or a combination of an MDM2 inhibitor and PC14586. [Figure 3B] Figures 3A-3H: MDM2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 3B: Dose-response survival curves of AML cells with TP53-WT treated for 72 hours with an MDM2 inhibitor, PC14586, or a combination of an MDM2 inhibitor and PC14586. [Figure 3C]Figures 3A-3H: MDM2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 3C: 7AAD / AnnV curves of AML cells with TP53-Y220C treated for 72 hours with an MDM2 inhibitor, PC14586, or a combination of an MDM2 inhibitor and PC14586. [Figure 3D] Figures 3A-3H: MDM2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 3D: Dose-response survival curves of AML cells with TP53-Y220C treated for 72 hours with an MDM2 inhibitor, PC14586, or a combination of an MDM2 inhibitor and PC14586. [Figure 3E] Figures 3A-3H: MDM2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 3E: 7AAD / AnnV curves of AML cells with TP53-KO treated for 72 hours with an MDM2 inhibitor, PC14586, or a combination of an MDM2 inhibitor and PC14586. [Figure 3F] Figures 3A-3H: MDM2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 3F: Dose-response survival curves of AML cells with TP53-KO treated for 72 hours with an MDM2 inhibitor, PC14586, or a combination of an MDM2 inhibitor and PC14586. [Figure 3G] Figures 3A-3H: MDM2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 3G: 7AAD / AnnV curves of AML cells with TP53-R175H treated for 72 hours with an MDM2 inhibitor, PC14586, or a combination of an MDM2 inhibitor and PC14586. [Figure 3H] Figures 3A-3H: MDM2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 3H: Dose-response survival curves of AML cells with TP53-R175H treated for 72 hours with an MDM2 inhibitor, PC14586, or a combination of an MDM2 inhibitor and PC14586. [Figure 4A] Figures 4A-4H: BCL-2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 4A: 7AAD / AnnV curves of AML cells with TP53-WT treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. [Figure 4B] Figures 4A-4H: BCL-2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 4B: Dose-response survival curves of AML cells with TP53-WT treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. [Figure 4C] Figures 4A-4H: BCL-2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 4C: 7AAD / AnnV curves of AML cells with TP53-Y220C treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. [Figure 4D]Figures 4A-4H: BCL-2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 4D: Dose-response survival curves of AML cells with TP53-Y220C treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. [Figure 4E] Figures 4A-4H: BCL-2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 4E: 7AAD / AnnV curves of AML cells with TP53-KO treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. [Figure 4F] Figures 4A-4H: BCL-2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 4F: Dose-response survival curves of AML cells with TP53-KO treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. [Figure 4G] Figures 4A-4H: BCL-2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 4G: 7AAD / AnnV curves of AML cells with TP53-R175H treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. [Figure 4H] Figures 4A-4H: BCL-2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 4H: Dose-response survival curves of AML cells with TP53-R175H treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. [Figure 5A] Figures 5A-5H: XPO-1 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 5A: 7AAD / AnnV curves of AML cells with TP53-WT treated for 72 hours with an XPO-1 inhibitor, PC14586, or a combination of an XPO-1 inhibitor and PC14586. [Figure 5B] Figures 5A-5H: XPO-1 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 5B: Dose-response survival curves of AML cells with TP53-WT treated for 72 hours with an XPO-1 inhibitor, PC14586, or a combination of an XPO-1 inhibitor and PC14586. [Figure 5C] Figures 5A-5H: XPO-1 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 5C: 7AAD / AnnV curves of AML cells with TP53-Y220C treated for 72 hours with an XPO-1 inhibitor, PC14586, or a combination of an XPO-1 inhibitor and PC14586. [Figure 5D] Figures 5A-5H: XPO-1 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 5D: Dose-response survival curves of AML cells with TP53-Y220C treated for 72 hours with an XPO-1 inhibitor, PC14586, or a combination of an XPO-1 inhibitor and PC14586. [Figure 5E]Figures 5A-5H: XPO-1 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 5E: 7AAD / AnnV curves of AML cells with TP53-KO treated for 72 hours with an XPO-1 inhibitor, PC14586, or a combination of an XPO-1 inhibitor and PC14586. [Figure 5F] Figures 5A-5H: XPO-1 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 5F: Dose-response survival curves of AML cells with TP53-KO treated for 72 hours with an XPO-1 inhibitor, PC14586, or a combination of an XPO-1 inhibitor and PC14586. [Figure 5G] Figures 5A-5H: XPO-1 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 5G: 7AAD / AnnV curves of AML cells with TP53-R175H treated for 72 hours with an XPO-1 inhibitor, PC14586, or a combination of an XPO-1 inhibitor and PC14586. [Figure 5H] Figures 5A-5H: XPO-1 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. Figure 5H: Dose-response survival curves of AML cells with TP53-R175H treated for 72 hours with an XPO-1 inhibitor, PC14586, or a combination of an XPO-1 inhibitor and PC14586. [Figure 6A]Figures 6A-6D: Regarding in vitro inhibition of AML cells with TP53-WT (WT) and AML cells with TP53-Y220C (Y220C), PC14586 combined with three other activators showed the best synergistic effect compared to PC14586 combined with one activator or PC14586 alone, followed by PC14586 combined with two other activators. Figure 6A: 7AAD / AnnV curves of AML cells with TP53-WT treated with PC14586, a BCL-2 inhibitor (BCL201), an MDM2 inhibitor (HDM201), an XPO-1 inhibitor (KPT-330), or a combination of PC14586 and one, two, or all three of these other activators after 72 hours of incubation. [Figure 6B] Figures 6A-6D: Regarding in vitro inhibition of AML cells with TP53-WT (WT) and AML cells with TP53-Y220C (Y220C), PC14586 combined with three other activators showed the best synergistic effect compared to PC14586 combined with one activator or PC14586 alone, followed by PC14586 combined with two other activators. Figure 6B: Dose-response survival curves of AML cells with TP53-WT treated with PC14586, a BCL-2 inhibitor, an MDM2 inhibitor, an XPO-1 inhibitor, or a combination of PC14586 and one, two, or all three of these other activators after 72 hours of incubation. [Figure 6C]Figures 6A-6D: Regarding in vitro inhibition of AML cells with TP53-WT (WT) and AML cells with TP53-Y220C (Y220C), PC14586 combined with three other activators showed the best synergistic effect compared to PC14586 combined with one activator or PC14586 alone, followed by PC14586 combined with two other activators. Figure 6C: 7AAD / AnnV curves of AML cells with TP53-Y220C treated with PC14586, a BCL-2 inhibitor, an MDM2 inhibitor, an XPO-1 inhibitor, or a combination of PC14586 and one, two, or all three of these other activators after 72 hours of incubation. [Figure 6D] Figures 6A-6D: Regarding in vitro inhibition of AML cells with TP53-WT (WT) and AML cells with TP53-Y220C (Y220C), PC14586 combined with three other activators showed the best synergistic effect compared to PC14586 combined with one activator or PC14586 alone, followed by PC14586 combined with two other activators. Figure 6D: Dose-response survival curves of AML cells with TP53-Y220C treated with PC14586, a BCL-2 inhibitor, an MDM2 inhibitor, an XPO-1 inhibitor, or a combination of PC14586 and one, two, or all three of these other activators after 72 hours of incubation. [Figure 7A]Figures 7A-7C: Molm13 TP53-Y220C cells treated with PC14586 alone or in combination with an XPO-1 inhibitor, MDM2 inhibitor, or BCL-2 inhibitor. Figure 7A: Molm13 treated for 24 hours with venetoclax (VEN, 5 nM or 10 nM), nutrin-3a (Nut, 2.5 μM or 5 μM), PC14586 (PC, 2 μM or 4 μM), KPT-8602 (KPT, 100 nM or 200 nM), or a combination of PC14586 and venetoclax (VEN / PC, 5 nM VEN and 2 μM PC, 10 nM VEN and 4 μM PC), a combination of PC14586 and nutrin-3a (Nut / PC, 2.5 μM Nut and 2 μM PC, 5 μM Nut and 4 μM PC), or a combination of PC14586 and KPT-8602 (KPT / PC, 100 nM KPT and 2 μM PC, 200 nM KPT and 4 μM PC). Western blot demonstrating protein levels in TP53-Y220C cells. [Figure 7B] Figures 7A-7C: Molm13 TP53-Y220C cells treated with PC14586 alone or in combination with an XPO-1 inhibitor, MDM2 inhibitor, or BCL-2 inhibitor. Figure 7B: Cell cycle distribution and apoptosis determined by flow cytometry of cells stained with 5-ethinyl-2'-deoxyuridine (EdU) and DNA dyes. Molm13 TP53-Y220C cells were treated for 72 hours with venetoclax (VEN, 10 nM), nutrin-3a (N3, 5 μM), KPT-8602 (KPT, 200 nM), PC14586 (4 μM), or a combination of PC14586 and venetoclax (4 μM PC14586 and 10 nM VEN), a combination of PC14586 and nutrin-3a (4 μM PC14586 and 5 μM N3), or a combination of PC14586 and KPT (4 μM PC14586 and 200 nM KPT). [Figure 7C]Figures 7A-7C: Molm13 TP53-Y220C cells treated with PC14586 alone or in combination with an XPO-1 inhibitor, MDM2 inhibitor, or BCL-2 inhibitor. Figure 7C: DNA content and PARP cleavage determined by flow cytometry of Molm13 TP53-Y220C cells treated for 72 hours with venetoclax (VEN, 10 nM), nutrin-3a (N3, 5 μM), KPT-8602 (KPT, 200 nM), PC14586 (4 μM), or a combination of PC14586 and venetoclax (4 μM PC14586 and 10 nM VEN), a combination of PC14586 and nutrin-3a (4 μM PC14586 and 5 μM), or a combination of PC14586 and KPT-8602 (4 μM PC14586 and 200 nM KPT). Cells were stained with EdU, DNA dyes, and antibodies against cleaved PARP. [Figure 8A] Figures 8A-8G: Cells derived from peripheral blood (PB) samples of AML patients, 77% of which had TP53-Y220C, under mesenchymal stromal cell (MSC) co-culture conditions (COX). Cells were stained with annexin V / 7-aminoactinomycin D (Ann V / 7-ADD) in the presence of counting beads, and cell death and viability were determined by flow cytometry. Figure 8A: CD45+, CD34+, and CD34+CD38- populations 48 hours after treatment with different concentrations of PC14586. [Figure 8B]Figures 8A-8G: Cells derived from peripheral blood (PB) samples of AML patients, 77% of which had TP53-Y220C, under mesenchymal stromal cell (MSC) co-culture conditions (COX). Cells were stained with annexin V / 7-aminoactinomycin D (Ann V / 7-ADD) in the presence of counting beads, and cell death and viability were determined by flow cytometry. Figure 8B: CD45+ population evaluated by Ann V / 7-ADD+ positive cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. [Figure 8C] Figures 8A-8G: Cells derived from peripheral blood (PB) samples of AML patients, 77% of which had TP53-Y220C, under mesenchymal stromal cell (MSC) co-culture conditions (COX). Cells were stained with annexin V / 7-aminoactinomycin D (Ann V / 7-ADD) in the presence of counting beads, and then cell death and viability were determined by flow cytometry. Figure 8C: CD45+ viable cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. [Figure 8D]Figures 8A-8G: Cells derived from peripheral blood (PB) samples of AML patients, 77% of which had TP53-Y220C, under mesenchymal stromal cell (MSC) co-culture conditions (COX). Cells were stained with annexin V / 7-aminoactinomycin D (Ann V / 7-ADD) in the presence of counting beads, and cell death and viability were determined by flow cytometry. Figure 8D: CD34+ cells evaluated by Ann V / 7ADD+ cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. [Figure 8E] Figures 8A-8G: Cells derived from peripheral blood (PB) samples of AML patients, 77% of which had TP53-Y220C, under mesenchymal stromal cell (MSC) co-culture conditions (COX). Cells were stained with annexin V / 7-aminoactinomycin D (Ann V / 7-ADD) in the presence of counting beads, and then cell death and viability were determined by flow cytometry. Figure 8E: CD34+ viable cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. [Figure 8F]Figures 8A-8G: Cells derived from peripheral blood (PB) samples of AML patients, 77% of which had TP53-Y220C, under mesenchymal stromal cell (MSC) co-culture conditions (COX). Cells were stained with annexin V / 7-aminoactinomycin D (Ann V / 7-ADD) in the presence of counting beads, and cell death and viability were determined by flow cytometry. Figure 8F: CD34+CD38- population evaluated by Ann V / 7ADD+ cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. [Figure 8G] Figures 8A-8G: Cells derived from peripheral blood (PB) samples of AML patients, 77% of which had TP53-Y220C, under mesenchymal stromal cell (MSC) co-culture conditions (COX). Cells were stained with annexin V / 7-aminoactinomycin D (Ann V / 7-ADD) in the presence of counting beads, and then cell death and viability were determined by flow cytometry. Figure 8G: CD34+CD38- viable cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. [Figure 9A] Figures 9A-9G: Cells derived from peripheral blood (PB) samples of AML patients, with 45.7% having TP53-Y220C, under MSC COX. Cell death and viability were determined by flow cytometry after staining with Ann V / 7-ADD in the presence of counting beads. Figure 9A: CD45+, CD34+, and CD34+CD38- populations 48 hours after treatment with different concentrations of PC14586. [Figure 9B]Figures 9A-9G: Cells derived from peripheral blood (PB) samples of AML patients with TP53-Y220C, under MSC COX. Cell death and viability were determined by flow cytometry after staining cells with Ann V / 7-ADD in the presence of counting beads. Figure 9B: CD45+ population evaluated by Ann V / 7-ADD+ positive cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. [Figure 9C] Figures 9A-9G: Cells derived from peripheral blood (PB) samples of AML patients with 45.7% TP53-Y220C under MSC COX. Cell death and viability were determined by flow cytometry after staining the cells with Ann V / 7-ADD in the presence of counting beads. Figure 9C: CD45+ viable cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. [Figure 9D] Figures 9A-9G: Cells derived from peripheral blood (PB) samples of AML patients with 45.7% having TP53-Y220C, under MSC COX. Cell death and viability were determined by flow cytometry after staining the cells with Ann V / 7-ADD in the presence of counting beads. Figure 9D: CD34+ evaluated by Ann V / 7ADD+ cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. [Figure 9E]Figures 9A-9G: Cells from peripheral blood (PB) samples of AML patients with 45.7% TP53-Y220C under MSC COX. Cell death and viability were determined by flow cytometry after staining cells with Ann V / 7-ADD in the presence of counting beads. Figure 9E: CD34+ viable cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. [Figure 9F] Figures 9A-9G: Cells derived from peripheral blood (PB) samples of AML patients with 45.7% having TP53-Y220C, under MSC COX. Cell death and viability were determined by flow cytometry after staining cells with Ann V / 7-ADD in the presence of counting beads. Figure 9F: CD34+CD38- population evaluated by Ann V / 7ADD+ cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. [Figure 9G] Figures 9A-9G: Cells derived from peripheral blood (PB) samples of AML patients with 45.7% having TP53-Y220C, under MSC COX. Cell death and viability were determined by flow cytometry after staining the cells with Ann V / 7-ADD in the presence of counting beads. Figure 9G: CD34+CD38- viable cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. [Figure 10A]Figures 10A-10D: Patient-derived xenograft cells with the Y220C mutation under MSC COX. Cells were stained with Ann V / 7-ADD in the presence of counting beads, and cell death and viability were determined by flow cytometry. Figure 10A: CD45+, CD34+, and CD34+CD38- populations 96 hours after treatment with different concentrations of PC14586. [Figure 10B] Figures 10A-10D: Patient-derived xenograft cells with the Y220C mutation under MSC COX. Cells were stained with Ann V / 7-ADD in the presence of counting beads, and cell death and viability were determined by flow cytometry. Figure 10B: CD45+ population evaluated by Ann V / 7-ADD+ positive cells 96 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; nutrin-3a, KPT-8602, and PC14586; venetoclax, KPT-8602, and PC14586; KPT-8602, nutrin-3a, and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586. [Figure 10C] Figures 10A-10D: Patient-derived xenograft cells with the Y220C mutation under MSC COX. Cells were stained with Ann V / 7-ADD in the presence of counting beads, and cell death and viability were determined by flow cytometry. Figure 10C: Venetoclax; Nutrin-3a (N3); KPT-8602; PC14586; Venetoclax and PC14586; Nutrin-3a and PC14586; KPT-8602 and PC14586; Nutrin-3a, KPT-8602, and PC14586; Venetoclax, KPT-8602, and PC14586; KPT-8602, Nutrin-3a, and PC14586; or CD45+ viable cells 96 hours after treatment with Venetoclax, Nutrin-3a, KPT-8602, and PC14586. [Figure 10D]Figures 10A-10D: Patient-derived xenograft cells with the Y220C mutation under MSC COX. Cells were stained with Ann V / 7-ADD in the presence of counting beads, and cell death and viability were determined by flow cytometry. Figure 10D: Venetoclax; Nutrin-3a (N3); KPT-8602; PC14586; Venetoclax and PC14586; Nutrin-3a and PC14586; KPT-8602 and PC14586; Nutrin-3a, KPT-8602, and PC14586; Venetoclax, KPT-8602, and PC14586; KPT-8602, Nutrin-3a, and PC14586; or CD34+ assessed by Ann V / 7ADD+ cells 96 hours after treatment with venetoclax, nutrin-3a, KPT-8602, and PC14586. [Figure 10E] Venetoclax; Nutrin-3a (N3); KPT-8602; PC14586; Venetoclax and PC14586; Nutrin-3a and PC14586; KPT-8602 and PC14586; Nutrin-3a, KPT-8602, and PC14586; Venetoclax, KPT-8602, and PC14586; KPT-8602, Nutrin-3a, and PC14586; or CD34+ viable cells 96 hours after treatment with Venetoclax, Nutrin-3a, KPT-8602, and PC14586. [Figure 10F] Venetoclax; Nutrin-3a (N3); KPT-8602; PC14586; Venetoclax and PC14586; Nutrin-3a and PC14586; KPT-8602 and PC14586; Nutrin-3a, KPT-8602, and PC14586; Venetoclax, KPT-8602, and PC14586; KPT-8602, Nutrin-3a, and PC14586; or CD34+CD38- population evaluated by Ann V / 7ADD+ cells 96 hours after treatment with venetoclax, nutrin-3a, KPT-8602, and PC14586. [Figure 10G]Venetoclax; Nutrin-3a (N3); KPT-8602; PC14586; Venetoclax and PC14586; Nutrin-3a and PC14586; KPT-8602 and PC14586; Nutrin-3a, KPT-8602, and PC14586; Venetoclax, KPT-8602, and PC14586; KPT-8602, Nutrin-3a, and PC14586; or CD34+CD38- viable cells 96 hours after treatment with Venetoclax, Nutrin-3a, KPT-8602, and PC14586. [Figure 11A] Figures 11A-11D: Normal bone marrow cells under MSC COX. Cells were stained with Ann V / 7-ADD in the presence of counting beads, and then cell death and viability were determined by flow cytometry. Figure 11A: CD45+ population evaluated by Ann V / 7-ADD+ positive cells 48 hours after treatment with venetoclax; nutrin-3a; KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586. [Figure 11B] Figures 11A-11D: Normal bone marrow cells under MSC COX. Cells were stained with Ann V / 7-ADD in the presence of counting beads, and cell death and viability were determined by flow cytometry. Figure 11B: CD45+ cell viability 48 hours after treatment with venetoclax; nutrin-3a; KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586. [Figure 11C]Figures 11A-11D: Normal bone marrow cells under MSC COX. Cells were stained with Ann V / 7-ADD in the presence of counting beads, and cell death and viability were determined by flow cytometry. Figure 11C: CD34+ population evaluated by Ann V / 7-ADD+ positive cells 48 hours after treatment with venetoclax; nutrin-3a; KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586. [Figure 11D] Figures 11A-11D: Normal bone marrow cells under MSC COX. Cells were stained with Ann V / 7-ADD in the presence of counting beads, and cell death and viability were determined by flow cytometry. Figure 11D: CD34+ cell viability 48 hours after treatment with venetoclax; nutrin-3a; KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586. [Figure 12A] Figures 12A-12F: Molm13 TP53-WT cells and TP53-Y220C cells were treated for 4 hours with Nutrin-3a (N3a, 5 μM), PC14586 (PC, 4 μM), or a combination thereof. RNA was isolated and subjected to RNA sequencing. Figure 12A: Principal component analysis. [Figure 12B] Figures 12A-12F: Molm13 TP53-WT cells and TP53-Y220C cells were treated for 4 hours with Nutrin-3a (N3a, 5 μM), PC14586 (PC, 4 μM), or a combination thereof. RNA was isolated and subjected to RNA sequencing. Figure 12B: Gene set enrichment analysis. [Figure 12C] Figures 12A-12F: Molm13 TP53-WT cells and TP53-Y220C cells were treated for 4 hours with nutrin-3a (N3a, 5 μM), PC14586 (PC, 4 μM), or a combination thereof. RNA was isolated and subjected to RNA sequencing. Figure 12C: Differently expressed genes. [Figure 12D] Figures 12A-12F: Molm13 TP53-WT cells and TP53-Y220C cells were treated for 4 hours with Nutrin-3a (N3a, 5 μM), PC14586 (PC, 4 μM), or a combination thereof. RNA was isolated and subjected to RNA sequencing. Figure 12D: Pathway analysis. [Figure 12E] Figures 12A-12F: Molm13 TP53-WT cells and TP53-Y220C cells were treated for 4 hours with Nutrin-3a (N3a, 5 μM), PC14586 (PC, 4 μM), or a combination thereof. RNA was isolated and subjected to RNA sequencing. Figure 12E: Volcano plots show comparisons of increased, unchanged, and decreased gene expression in treated cells compared to controls. [Figure 12F] Figures 12A-12F: Molm13 TP53-WT cells and TP53-Y220C cells were treated for 4 hours with Nutrin-3a (N3a, 5 μM), PC14586 (PC, 4 μM), or a combination thereof. RNA was isolated and subjected to RNA sequencing. Figure 12F: Western blot showing changes in several gene expressions. [Figure 13A] Figures 13A-13C: Patient-derived xenograft cells from AML patient samples bearing the TP53-Y220C (VAF 48%) mutation, TP53-P151A (VAF 47%) mutation, and NRAS (VAF 50%) mutation were injected into NSG mice via the tail vein and subsequently subjected to treatment. Treatments included a vehicle (negative control), PC14586 (100 mg / kg, daily) and the 50% active MDM2 inhibitor idasanutrin (RG7388) (40 mg / kg, initially 8 days on, then 2 days off / 5 days on), or both PC14586 (100 mg / kg, daily) and the 50% active MDM2 inhibitor idasanutrin (RG7388) (40 mg / kg, initially 8 days on, then 2 days off / 5 days on). Figure 13A: Flow cytometry analysis of human CD45+ cells in peripheral blood (PB) 4 weeks after treatment. [Figure 13B]Figures 13A-13C: Patient-derived xenograft cells from AML patient samples bearing the TP53-Y220C (VAF 48%) mutation, TP53-P151A (VAF 47%) mutation, and NRAS (VAF 50%) mutation were injected into NSG mice via the tail vein and subsequently subjected to treatment. Treatments included a vehicle (negative control), PC14586 (100 mg / kg, daily) and the 50% active MDM2 inhibitor idasanutrin (RG7388) (40 mg / kg, initially 8 days on, then 2 days off / 5 days on), or both PC14586 (100 mg / kg, daily) and the 50% active MDM2 inhibitor idasanutrin (RG7388) (40 mg / kg, initially 8 days on, then 2 days off / 5 days on). Figure 13B: Flow cytometry analysis of human CD45+ cells in peripheral blood (PB) 8 weeks after treatment. [Figure 13C] Figures 13A-13C: Patient-derived xenograft cells from AML patient samples bearing the TP53-Y220C (VAF 48%) mutation, TP53-P151A (VAF 47%) mutation, and NRAS (VAF 50%) mutation were injected into NSG mice via the tail vein and subsequently subjected to treatment. Treatments included a vehicle (negative control), PC14586 (100 mg / kg, daily) and the 50% active MDM2 inhibitor idasanutrin (RG7388) (40 mg / kg, initially 8 days on, then 2 days off / 5 days on), or both PC14586 (100 mg / kg, daily) and the 50% active MDM2 inhibitor idasanutrin (RG7388) (40 mg / kg, initially 8 days on, then 2 days off / 5 days on). Figure 13C: Survival data analyzed using Kaplan-Meier estimation and log-rank test for mouse survival time. [Figure 14] Western blots of p53 and BCL-2 derived from Molm13 TP53-WT cells and TP53-Y220C cells treated with Nutrin-3a (5 μM) or PC14586 (PC, 1.25 μM or 5 μM) for 4 hours. [Figure 15]Survival data of mice treated daily with Molm13 TP53-Y220C cells and either vehicle, PC14586 (100 mg / kg), venetoclax (50 mg / kg), or a combination thereof, analyzed using Kaplan-Meier estimation and log-rank tests. [Figure 16] Western blots of p53, lamin B1, and tubulin derived from TP53-Y220C Molm13 cells treated with PC14586 (4 μM), KPT-8602 (200 nM), or both for 24 hours. [Modes for carrying out the invention]

[0020] Detailed explanation To provide a substantial understanding of this technology, certain aspects, forms, embodiments, variations, and characteristics of this method are described below at various levels of detail. This disclosure is not limited to any particular use, method, reagent, compound, composition, or biological system, and these are, of course, modifiable. The terminology used herein is for illustrative purposes only and is not intended to limit it to any particular aspect.

[0021] In carrying out this method, many conventional techniques from molecular biology, protein biochemistry, cell biology, immunology, microbiology, and recombinant DNA are used. For example, Sambrook and Russell eds. (2001) Molecular Cloning: A Laboratory Manual, 3rd edition; the series Ausubel et al. eds. (2007) Current Protocols in Molecular Biology; the series Methods in Enzymology (Academic Press, Inc., NY); MacPherson et al. (1991) PCR 1: A Practical Approach (IRL Press at Oxford University Press); MacPherson et al. (1995) PCR 2: A Practical Approach;Harlow and Lane eds. (1999) Antibodies, A Laboratory Manual;Freshney (2005) Culture of Animal Cells: A Manual of Basic Technique, 5th edition;Gait ed. (1984) Oligonucleotide Synthesis;US Patent No. 4,683,195;Hames and Higgins eds. (1984) Nucleic Acid Hybridization;Anderson (1999) Nucleic Acid Hybridization;Hames and Higgins eds. (1984) Transcription and Translation;Immobilized Cells and Enzymes (IRL Press (1986));Perbal (1984) A Practical Guide to Molecular Cloning;Miller and Calos eds.See (1987) Gene Transfer Vectors for Mammalian Cells (Cold Spring Harbor Laboratory); Makrides ed. (2003) Gene Transfer and Expression in Mammalian Cells; Mayer and Walker eds. (1987) Immunochemical Methods in Cell and Molecular Biology (Academic Press, London); and Herzenberg et al. eds (1996) Weir's Handbook of Experimental Immunology. Methods for detecting and measuring polypeptide gene expression levels (i.e., gene translation levels) are well known in the art, including the use of polypeptide detection methods such as antibody detection and quantification techniques. (See also Strachan & Read, Human Molecular Genetics, Second Edition. (John Wiley and Sons, Inc., NY, 1999)).

[0022] definition Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which this art belongs. Where used herein and in the appended claims, the singular forms "a," "an," and "the" include plural nouns unless otherwise explicitly indicated. For example, a reference to "a cell" includes combinations of two or more cells and similar combinations. Generally, the nomenclature used herein, as well as the laboratory procedures in cell culture, molecular genetics, organic chemistry, analytical chemistry, nucleic acid chemistry, and hybridization described below, are well known and commonly used in the art.

[0023] When used in this disclosure, the phrase "and / or" will be understood to mean either any one of the members described individually, or any combination of two or more of them. For example, "A, B, and / or C" means "A, B, C, A and B, A and C, B and C, or any combination of A, B, and C."

[0024] Generally, a reference to a particular element, such as hydrogen or H, is intended to include all isotopes of that element. For example, if the R group is defined as containing hydrogen or H, then it also includes deuterium and tritium. Thus, tritium, C 14 , P 32 , and S 35 Compounds containing radioactive isotopes such as those mentioned above are within the scope of this technology. Procedures for inserting such labels into compounds of this technology will be readily apparent to those skilled in the art based on the disclosures herein.

[0025] As used herein, the term “about” with respect to numbers generally means, unless otherwise stated or otherwise evident from the context, a number that falls within 1%, 5%, or 10% in either direction (greater than or less than) of the number (except where such a number falls less than 0% or greater than 100% of the possible value).

[0026] As used herein, “administration” of an active substance or drug to a subject includes any route through which the compound is introduced or delivered to the subject to perform its intended function. Administration may be carried out by any preferred route, including but not limited to oral, intranasal, intrathecal, parenteral (intravenous, intramuscular, intraperitoneal, or subcutaneous), transrectal, intrathecal, intraocular, intradermal, transmucosal, iontophoresis, or topical. Administration includes self-administration and administration by a non-self-administered person.

[0027] As used herein, the terms “cancer” and “tumor” are interchangeable and refer to the presence of cells that have characteristics typical of cancer-causing cells, such as uncontrolled growth, immortality, metastatic ability, rapid proliferation and growth rate, and certain characteristic morphological features. While cancer cells often take the form of tumors, such cells can exist alone in animals or can be non-tumor-forming cancer cells. As used herein, the term “cancer cells” includes precancerous (e.g., benign) cells, malignant cells, premetastatic cells, metastatic cells, and nonmetastatic cells. Cancers of virtually any tissue are known to those skilled in the art, including solid tumors such as carcinomas, sarcomas, glioblastomas, and melanomas, and circulating cancers such as leukemia. Examples of cancers include, but are not limited to, ovarian cancer, breast cancer, colon cancer, lung cancer, prostate cancer, stomach cancer, pancreatic cancer, cervical cancer, liver cancer, bladder cancer, urinary tract cancer, thyroid cancer, kidney cancer, carcinomas, melanomas, head and neck cancers, and brain cancers. The terms "cancer load" or "tumor load" refer to the quantity of cancer cells or tumor volume in a subject. Consequently, reducing the cancer load may refer to reducing the number of cancer cells or tumor volume in a subject. The term "cancer cell" refers to cells that exhibit cancer-like characteristics, such as uncontrolled replication, resistance to antiproliferative signals, ability to metastasize, and loss of the ability to undergo programmed cell death (e.g., apoptosis), or cells derived from cancer cells, such as clones of cancer cells.

[0028] As used herein, “control” refers to a substitute sample used in an experiment for comparative purposes. A control may be “positive” or “negative.” For example, if the purpose of the experiment is to determine the correlation of the potency of therapeutic substances for the treatment of a particular type of disease, a positive control (a compound or composition known to exhibit the desired therapeutic effect) and a negative control (a subject or sample that receives no treatment or is given a placebo) are typically used.

[0029] As used herein, the term “effective dose” means an amount sufficient to achieve the desired therapeutic and / or preventive effect, for example, an amount that results in the inhibition or reduction of one or more signs or symptoms of the disease or condition described herein or associated with the disease or condition described herein. In the context of therapeutic or preventive use, the amount of composition administered to a subject will vary depending on the composition, the degree, type, and severity of the disease, as well as individual characteristics such as overall health, age, sex, weight, and drug tolerance. A person skilled in the art can determine the appropriate dosage in accordance with these and other factors. The composition may also be administered in combination with one or more additional therapeutic compounds. In the methods described herein, the therapeutic composition may be administered to a subject having one or more signs or symptoms of the disease or condition described herein. As used herein, the “effective dose” of the composition means the level of composition at which the physiological effects of the disease or condition are remitted or eliminated. The effective dose may be induced in one or more doses.

[0030] As used herein, “expression” includes one or more of the following: transcription of a gene into an mRNA precursor; splicing and other processing of the mRNA precursor to produce mature mRNA; mRNA stability; translation of mature mRNA into a protein (including codon use and tRNA availability); and glycosylation and / or other modifications of the translation product, if necessary for appropriate expression and function.

[0031] As used herein, the terms “polypeptide,” “peptide,” and “protein” are used interchangeably to mean polymers containing two or more amino acids linked together by peptide bonds, or modified peptide bonds, i.e., peptide equivalents. Polypeptides refer to both short chains, commonly called peptides, glycopeptides, or oligomers, and longer chains, commonly called proteins. Polypeptides may contain amino acids other than the 20 gene-coding amino acids. Polypeptides contain amino acid sequences modified by either natural processes such as post-translational processing or chemical modification techniques well known in the art.

[0032] As used herein, “sample” or “biological sample” refers to a fluid or tissue sample isolated from a subject. In some cases, a biological sample may consist of or include whole blood, platelets, erythrocytes, leukocytes, plasma, serum, urine, feces, epidermal samples, vaginal samples, skin samples, oral swabs, sperm, amniotic fluid, cultured cells, bone marrow samples, tumor biopsies, chorionic villus aspirates and / or chorionic villi, cultured cells, endothelial cells, synovial fluid, lymph, ascites, interstitial fluid or extracellular fluid, and similar substances. The term “sample” may also include fluids in the intercellular spaces, including gingival crevicular exudate, bone marrow, cerebrospinal fluid (CSF), saliva, mucus, spit, semen, sweat, urine, or any other body fluid. Samples can be obtained from a subject by any means, including but not limited to venous puncture, excretion, ejaculation, massage, biopsy, needle aspiration, washing, scraping, surgical incision, or intervention, or other means known in the art. Blood samples may be whole blood or any fraction thereof, including blood cells (red blood cells, white blood cells or leukocytes, and platelets), serum, and plasma.

[0033] As used herein, the term “separate” therapeutic use refers to the administration of at least two active ingredients by different routes at the same or substantially the same time.

[0034] As used herein, the term “sequential” therapeutic use refers to the administration of at least two active ingredients at different times. More specifically, sequential use means that the administration of one or the other active ingredient is initiated after the entire administration of one of the active ingredients. Thus, it is possible to administer one of the active ingredients over several minutes, hours, or days, and then administer the other or other active ingredient. In this case, there is no simultaneous treatment.

[0035] As used herein, the term “concurrent” therapeutic use means the administration of at least two active ingredients by the same route at the same or substantially the same time.

[0036] As used herein, the terms “subject,” “patient,” or “individual” may refer to an individual organism, such as a vertebrate, mammal, or human. In some embodiments, the subject, patient, or individual is a human.

[0037] As used herein, “synergistic therapeutic effect” refers to an additive therapeutic effect produced by a combination of at least two active ingredients that exceeds what would normally be produced by individual doses of the active ingredients. For example, lower doses of one or more active ingredients may be used to treat a disease or disorder.

[0038] As used herein, the term “therapeutic substance” is intended to mean a compound that, when present in an effective amount, imparts a desired therapeutic effect to a subject in need.

[0039] When used herein, “treatment” or “treatment” covers treatment of a disease or disorder described herein in a subject such as a human, and includes (i) inhibiting the disease or disorder, i.e., preventing its onset; (ii) alleviating the disease or disorder, i.e., causing regression of the disorder; (iii) slowing the exacerbation of the disorder; and / or (iv) inhibiting, alleviating, or slowing the exacerbation of one or more symptoms of the disease or disorder. In some embodiments, treatment means, for example, reducing, decreasing, curing, or improving the symptoms associated with the disease.

[0040] It will also be recognized that the various forms of treatment for disorders described herein, including but not limited to comprehensive treatments, are intended to mean “substantial” treatments in which some biological or medically relevant outcome is achieved. Treatment may be a continuous, long-term treatment for a chronic disease, or a single or multiple administration for the treatment of an acute condition.

[0041] Generally, a reference to a particular element, such as hydrogen or H, is intended to include all isotopes of that element. For example, if the R group is defined as containing hydrogen or H, then it also includes deuterium and tritium. Thus, tritium, C 14 , P 32 , and S 35 Compounds containing radioactive isotopes such as those mentioned above are within the scope of this technology. Procedures for inserting such labels into compounds of this technology will be readily apparent to those skilled in the art based on the disclosures herein.

[0042] Generally, "substituted" refers to an organic group (e.g., an alkyl group) in which one or more bonds to a hydrogen atom are replaced by bonds to a non-hydrogen or non-carbon atom. Substituted groups also include those in which one or more bonds to a carbon or hydrogen atom are replaced by one or more bonds to a heteroatom, including double or triple bonds. Therefore, unless otherwise specified, substituted groups are substituted with one or more substituents. In some embodiments, substituted groups are substituted with one, two, three, four, five, or six substituents. Examples of substituents include halogens (i.e., F, Cl, Br, and I); hydroxyl; alkoxy groups, alkenoxy groups, aryloxy groups, aralkyloxy groups, heterocyclyl groups, heterocyclylalkyl groups, heterocyclyloxy groups, and heterocyclylalkoxy groups; carbonyl (oxo); carboxylates; esters; urethanes; oximes; hydroxylamines; alkoxyamines; aralkoxyamines; thiols; sulfides; sulfoxides; sulfones; sulfonyls; pentafluorosulfanyl (i.e., SF5), sulfonamides; amines; N-oxides; hydrazines; hydrazides; hydrazones; azides; amides; ureas; amidines; guanidines; enamines; imides; isocyanates; isothiocyanates; cyanates; thiocyanates; imines; nitro groups; and nitriles (i.e., CN).

[0043] Substituted ring groups, such as substituted cycloalkyl groups, aryl groups, heterocyclyl groups, and heteroaryl groups, also include rings and ring systems in which bonds to hydrogen atoms are replaced by bonds to carbon atoms. Therefore, substituted cycloalkyl groups, aryl groups, heterocyclyl groups, and heteroaryl groups may also be substituted with substituted or unsubstituted alkyl groups, alkenyl groups, and alkynyl groups as defined below.

[0044] Alkyl groups include linear and branched alkyl groups having 1 to 12 carbon atoms, typically 1 to 10 carbon atoms, or in some embodiments, 1 to 8, 1 to 6, or 1 to 4 carbon atoms. Alkyl groups can be substituted or unsubstituted. Examples of linear alkyl groups include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, isobutyl, sec-butyl, tert-butyl, neopentyl, isopentyl, and 2,2-dimethylpropyl groups. Typical substituted alkyl groups may be one or more times substituted with substituents such as those listed above, and typical substituted alkyl groups include, without limitation, haloalkyls (e.g., trifluoromethyl), hydroxyalkyls, thioalkyls, aminoalkyls, alkylaminoalkyls, dialkylaminoalkyls, alkoxyalkyls, carboxyalkyls, and the like.

[0045] Cycloalkyl groups include monocyclic, bicyclic, or tricyclic alkyl groups having 3 to 12 carbon atoms in the ring, or in some embodiments, 3 to 10, 3 to 8, or 3 to 4, 5, or 6 carbon atoms. Cycloalkyl groups may be substituted or unsubstituted. Exemplary monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some embodiments, cycloalkyl groups have 3 to 8 ring members, while in other embodiments, the number of ring carbon atoms ranges from 3 to 5, 3 to 6, or 3 to 7. Bicyclic and tricyclic ring systems include, but are not limited to, bicyclo[2.1.1]hexane, adamantyl, dekalinyl, and similar groups, encompassing both cross-linked cycloalkyl groups and fused rings. Substituted cycloalkyl groups may be one or more times substituted with non-hydrogen and non-carbon groups as defined above. On the other hand, substituted cycloalkyl groups also include rings substituted with linear or branched alkyl groups as defined above. Typical substituted cycloalkyl groups are monosubstituted or substituted with substituents such as those listed above, including but not limited to, 2,2-, 2,3-, 2,4-, 2,5-, or 2,6-disubstituted cyclohexyl groups, which may be one-time hypersubstituted.

[0046] A cycloalkylalkyl group is an alkyl group as defined above, in which a hydrogen bond or carbon bond of the alkyl group is replaced by a bond to the cycloalkyl group as defined above. Cycloalkylalkyl groups can be substituted or unsubstituted. In some embodiments, cycloalkylalkyl groups have 4 to 16 carbon atoms, 4 to 12 carbon atoms, and typically 4 to 10 carbon atoms. A substituted cycloalkylalkyl group may have substitution in the alkyl moiety, the cycloalkyl moiety, or both the alkyl and cycloalkyl moieties of the group. Typical substituted cycloalkylalkyl groups are monosubstituted, or may be monosubstituted, disubstituted, or trisubstituted with substituents such as those listed above, and may be once-supersubstituted.

[0047] Alkenyl groups include the linear and branched alkyl groups defined above, except that they have at least one double bond between two carbon atoms. Alkenyl groups can be substituted or unsubstituted. Alkenyl groups have 2 to 12 carbon atoms, typically 2 to 10 carbon atoms, or in some embodiments, 2 to 8, 2 to 6, or 2 to 4 carbon atoms. In some embodiments, alkenyl groups have one, two, or three carbon-carbon double bonds. Examples include, but are not limited to, vinyl, allyl, -CH=CH(CH3), -CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), and -C(CH2CH3)=CH2. Typical substituted alkenyl groups are monosubstituted or can be monosubstituted, disubstituted, or trisubstituted with substituents such as those listed above, but are not limited to, and may be once hypersubstituted.

[0048] Cycloalkenyl groups include cycloalkyl groups as defined above, which have at least one double bond between two carbon atoms. Cycloalkenyl groups may be substituted or unsubstituted. In some embodiments, cycloalkenyl groups may have one, two, or three double bonds, but may not contain aromatic compounds. Cycloalkenyl groups have 4 to 14 carbon atoms, or in some embodiments, 5 to 14 carbon atoms, 5 to 10 carbon atoms, or even 5, 6, 7, or 8 carbon atoms. Examples of cycloalkenyl groups include cyclohexenyl, cyclopentenyl, cyclohexadienyl, cyclobutadienyl, and cyclopentadienyl.

[0049] A cycloalkenylalkyl group is an alkyl group as defined above, in which the hydrogen or carbon bond of the alkyl group is replaced by a bond to the cycloalkenyl group as defined above. Cycloalkenylalkyl groups can be substituted or unsubstituted. A substituted cycloalkenylalkyl group may be substituted in the alkyl moiety, the cycloalkenyl moiety, or both the alkyl and cycloalkenyl moieties of the group. Typical substituted cycloalkenylalkyl groups may be substituted once or multiple times with substituents, such as those listed above.

[0050] Alkynyl groups include the linear and branched alkyl groups defined above, except that they have at least one triple bond between two carbon atoms. Alkynyl groups can be substituted or unsubstituted. Alkynyl groups have 2 to 12 carbon atoms, typically 2 to 10 carbon atoms, or in some embodiments, 2 to 8, 2 to 6, or 2 to 4 carbon atoms. In some embodiments, alkynyl groups have one, two, or three carbon-carbon triple bonds. Examples include, but are not limited to, -C≡CH, -C≡CCH3, -CH2C≡CCH3, and -C≡CCH2CH(CH2CH3)2. Typical substituted alkynyl groups are monosubstituted or can be monosubstituted, disubstituted, or trisubstituted with substituents such as those listed above, but are not limited to, and can be once hypersubstituted.

[0051] Aryl groups are cyclic aromatic hydrocarbons that do not contain heteroatoms. Aryl groups as used herein include monocyclic, bicyclic, and tricyclic systems. Aryl groups may be substituted or unsubstituted. Therefore, aryl groups include, but are not limited to, phenyl, azlenyl, heptalenyl, biphenyl, fluorenyl, phenantrenyl, anthracenyl, indenyl, indanyl, pentarenyl, and naphthyl groups. In some embodiments, aryl groups contain 6 to 14 carbon atoms in the ring portion of the group, in other cases 6 to 12, or even 6 to 10 carbon atoms. In some embodiments, aryl groups are phenyl or naphthyl. The term "aryl group" includes groups containing fused rings, such as fused aromatic aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, and similar). Typical substituted aryl groups can be monosubstituted (e.g., tolyls) or once hypersubstituted. For example, monosubstituted aryl groups include, but are not limited to, 2-, 3-, 4-, 5-, or 6-substituted phenyl or naphthyl groups, which may be substituted with substituents such as those listed above.

[0052] An aralkyl group is an alkyl group as defined above, in which a hydrogen bond or carbon bond of the alkyl group is replaced by a bond to an aryl group as defined above. An aralkyl group may be substituted or unsubstituted. In some embodiments, an aralkyl group contains 7 to 16 carbon atoms, 7 to 14 carbon atoms, or 7 to 10 carbon atoms. A substituted aralkyl group may be substituted in the alkyl moiety, the aryl moiety, or both the alkyl and aryl moieties. Typical aralkyl groups include, but are not limited to, the benzyl and phenethyl groups, as well as condensed (cycloalkylaryl) alkyl groups such as 4-indanylethyl. Typical substituted aralkyl groups may be one or more times substituted with substituents such as those listed above.

[0053] Heterocyclyl groups include aromatic (also called heteroaryl) and non-aromatic ring compounds that contain three or more ring members, one or more of which are heteroatoms such as, but not limited to, N, O, and S. Heterocyclyl groups may be substituted or unsubstituted. In some embodiments, heterocyclyl groups contain one, two, three, or four heteroatoms. In some embodiments, heterocyclyl groups include monocyclic, bicyclic, and tricyclic rings having 3 to 16 ring members, while other such groups have 3 to 6, 3 to 10, 3 to 12, or 3 to 14 ring members. Heterocyclyl groups include aromatic ring systems such as imidazolyl, imidazolinyl, and imidazolidinyl groups, partially unsaturated ring systems, and saturated ring systems. The term "heterocyclyl group" refers to, for example, benzotriazolyl, 2,3-dihydrobenzo[1,4]dioxynyl, and benzo[1,3] Includes condensed ring species, including those containing condensed aromatic groups and condensed non-aromatic groups such as dioxoyl. The term also includes, but is not limited to, quinuclidyl, and bridging polycyclic ring systems containing heteroatoms. The term includes heterocyclyl groups having other groups such as alkyl groups, oxo groups, or halo groups bonded to one of the ring members, referred to as "substituted heterocyclyl groups." Heterocyclyl groups include azilidinyl group, azetidinyl group, pyrrolidinyl group, imidazolidinyl group, pyrazolidinyl group, thiazolidinyl group, tetrahydrothiophenyl group, tetrahydrofuranyl group, dioxolyl group, furanyl group, thiophenyl group, pyrrolyl group, pyrrolinyl group, imidazolyl group, imidazolinyl group, pyrazolyl group, pyrazolinyl group, triazolyl group, tetrazolyl group, oxazolyl group, isoxazolyl group, thiazolyl group, thiazolinyl group, isothiazolyl group, thiadiazolyl group, oxadiazolyl group, piperidyl group, piperazinyl group, morpholinyl group, thiomorpholinyl group, tetrahydropyranyl group, tetrahydrothiopyranyl group, oxatian group, and dioxyl. Group, dithianyl group, pyranyl group, pyridyl group, pyrimidinyl group, pyridadinyl group, pyrazinyl group, triazinyl group, dihydropyridyl group, dihydrodithinyl group, dihydrodithionyl group, homopiperazinyl group, quinuclidyl group, indolyl group, indolinyl group, isoindolyl group, azaindolyl(pyrrolopyridyl) group, indazolyl group, indolidinyl group, benzotriazolyl group, benzimidazolyl group, benzofuranyl group, benzothiophenyl group, benzothiazolyl group, benzoxadiazolyl group, benzoxazinyl group, benzodithinyl group, benzoxadinyl group, benzoxazolyl group, benzothiazolyl group, benzothiadiazolyl group, benzo[1,3] Dioxolyl group, pyrazolopyridyl group, imidazopyridyl (azabenzoimidazolyl) group, triazolopyridyl group, isoxazolopyridyl group, purinyl group, xanthinyl group, adeninyl group, guaninyl group, quinolinyl group, isoquinolinyl group, quinolidinyl group, quinoxalinyl group, quinazolinyl group, sinnolinyl group, phthalazinyl group, naphthylidinyl group, pteridinyl group, thianaphthyl group, dihydrobenzothiadinyl group, dihydrobe These include, but are not limited to, the dinzofuranyl group, dihydroindolyl group, dihydrobenzodioxynyl group, tetrahydroindolyl group, tetrahydroindazolyl group, tetrahydrobenzimidazolyl group, tetrahydrobenzotriazolyl group, tetrahydropyrrolopyridyl group, tetrahydropyrazolopyridyl group, tetrahydroimidazopyridyl group, tetrahydrotriazolopyridyl group, and tetrahydroquinolinyl group. Typical substituted heterocyclyl groups are monosubstituted, or 2-, 3-, 4-, 5-, or 6-substituted with various substituents such as those listed above, or disubstituted pyridyl or morpholinyl groups, but are not limited to these, and may also be once-supersubstituted.

[0054] A heteroaryl group is an aromatic ring compound containing five or more ring members, one or more of which are heteroatoms such as, but not limited to, N, O, and S. Heteroaryl groups can be substituted or unsubstituted. Examples of heteroaryl groups include pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, pyridadinyl, pyrimidinyl, pyrazinyl, thiophenyl, benzothiophenyl, furanyl, benzofuranyl, indolyl, azaindolyl (pyrrolopyridinyl), indazolyl, benzimidazolyl, imidazopyridinyl (azabenzimidazolyl), and pyrazolyl. Heteroaryl groups include, but are not limited to, groups such as zolopyridinyl, triazolopyridinyl, benzotriazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthyl, prinyl, xanthinyl, adeninyl, guanyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl. Heteroaryl groups include fused ring compounds in which all rings are aromatic, such as indolyl groups, and fused ring compounds in which only one of the rings is aromatic, such as 2,3-dihydroindolyl groups. Typical substituted heteroaryl groups may be one or more times substituted with various substituents, such as those listed above.

[0055] A heterocyclylalkyl group is an alkyl group as defined above, in which a hydrogen bond or carbon bond of the alkyl group is replaced by a bond to a heterocyclyl group as defined above. Heterocyclylalkyl groups can be substituted or unsubstituted. A substituted heterocyclylalkyl group may have substitution in the alkyl moiety, the heterocyclyl moiety, or both the alkyl and heterocyclyl moieties of the group. Representative heterocyclylalkyl groups include, but are not limited to, morpholine-4-ylethyl, furan-2-ylmethyl, imidazole-4-ylmethyl, pyridine-3-ylmethyl, tetrahydrofuran-2-ylethyl, and indole-2-ylpropyl. Representative substituted heterocyclylalkyl groups may have one or more substituents, such as those listed above.

[0056] A heteroaralkyl group is an alkyl group as defined above, in which a hydrogen bond or carbon bond of the alkyl group is replaced by a bond to a heteroaryl group as defined above. Heteroaralkyl groups can be substituted or unsubstituted. A substituted heteroaralkyl group may be substituted in the alkyl moiety, the heteroaryl moiety, or both the alkyl and heteroaryl moieties. Typical substituted heteroaralkyl groups may be substituted once or multiple times with substituents such as those listed above.

[0057] Groups described herein that have two or more addition points (i.e., divalent, trivalent, or polyvalent) in the compounds of this technology are designated by the use of the suffix "ene". For example, a divalent alkyl group is an alkylene group, a divalent aryl group is an arylene group, a divalent heteroaryl group is a divalent heteroarylene group, and so on. Substituted groups having a single addition point to the compounds of this technology are not referred to using the "ene" designation. Therefore, for example, chloroethyl is not referred to as chloroethylene herein.

[0058] An alkoxy group is a hydroxyl group (-OH) in which a bond to a hydrogen atom is replaced by a bond to a carbon atom of a substituted or unsubstituted alkyl group as defined above. Alkoxy groups can be substituted or unsubstituted. Examples of linear alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, pentoxy, hexoxy, and similar groups. Examples of branched alkoxy groups include, but are not limited to, isopropoxy, sec-butoxy, tert-butoxy, isopentoxy, isohexoxy, and similar groups. Examples of cycloalkoxy groups include, but are not limited to, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and similar groups. Typical substituted alkoxy groups may be one or more substituted with substituents such as those listed above.

[0059] The terms "alkyloyl" and "alkyloyloxy," as used herein, may refer to a -C(O)-alkyl group and an -OC(O)-alkyl group, respectively. Similarly, "aryloyl" and "aryloyloxy" may refer to a -C(O)-aryl group and an -OC(O)-aryl group.

[0060] The terms "aryloxy" and "arylalkoxy" refer to substituted or unsubstituted aryl groups and alkyl groups, respectively, that are bonded to an oxygen atom. Examples include, but are not limited to, phenoxy, naphthyloxy, and benzyloxy. Typical substituted aryloxy and arylalkoxy groups may be one or more substituted with substituents, such as those listed above.

[0061] As used herein, the term "carboxylate" refers to the -COOH group.

[0062] The term "ester" is used herein with respect to -COOR 70 This refers to the group and the -C(O)OG group. 70 is a substituted or unsubstituted alkyl group, cycloalkyl group, alkenyl group, alkynyl group, aryl group, aralkyl group, heterocyclylalkyl group, or heterocyclyl group as defined herein. G is a carboxylate protecting group. Carboxylate protecting groups are well known to those skilled in the art. A comprehensive list of protecting groups for carboxylate functionalization can be found in Protective Groups in Organic Synthesis, Greene, TW; Wuts, PGM, John Wiley & Sons, New York, NY, (3rd Edition, 1999), which can be added or removed using the procedures described therein, and this document is incorporated herein by reference in its entirety for any purpose as it is fully presented herein.

[0063] The term "amide" (or "amido") refers to the C-amide group and the N-amide group, i.e., -C(O)NR, respectively. 71 R 72 Base and -NR 71 C(O)R 72 Includes the group R 71 and R 72 This is independently hydrogen, or a substituted or unsubstituted alkyl group, alkenyl group, alkynyl group, cycloalkyl group, aryl group, aralkyl group, heterocyclylalkyl group, or heterocyclyl group as defined herein. Therefore, the amide group includes, but is not limited to, a carbamoyl group (-C(O)NH2) and a formamide group (-NHC(O)H). In some embodiments, the amide is -NR 71 C(O)-(C 1~5In other cases, the amide is -NHC(O)-alkyl, and this group is called "carbonylamino."

[0064] The terms "nitrile" or "cyano" as used herein refer to the -CN group.

[0065] The urethane group consists of N-urethane groups and O-urethane groups, i.e., -NR groups, respectively. 73 C(O)OR 74 Base and -OC(O)NR 73 R 74 It contains the group R 73 and R 74 R is independently a substituted or unsubstituted alkyl group, alkenyl group, alkynyl group, cycloalkyl group, aryl group, aralkyl group, heterocyclylalkyl group, or heterocyclyl group as defined herein. 73 It could also be H.

[0066] The term "amine" (or "amino") is used herein by -NR 75 R 76 It refers to the base, R 75 and R 76 This is independently hydrogen, or a substituted or unsubstituted alkyl group, alkenyl group, alkynyl group, cycloalkyl group, aryl group, aralkyl group, heterocyclylalkyl group, or heterocyclyl group as defined herein. In some embodiments, the amine is an alkylamino, dialkylamino, arylamino, or alkylarylamino. In other embodiments, the amine is NH2, methylamino, dimethylamino, ethylamino, diethylamino, propylamino, isopropylamino, phenylamino, or benzylamino.

[0067] The term "sulfonamide" refers to the S-sulfonamide group and the N-sulfonamide group, i.e., -SO2NR, respectively. 78 R 79Base and -NR 78 SO2R 79 Includes the group R 78 and R 79 This group is independently hydrogen, or a substituted or unsubstituted alkyl group, alkenyl group, alkynyl group, cycloalkyl group, aryl group, aralkyl group, heterocyclylalkyl group, or heterocyclyl group as defined herein. Therefore, sulfonamide groups include, but are not limited to, sulfamoyl (-SO2NH2) groups. In some embodiments herein, the sulfonamide is -NHSO2-alkyl and is referred to as an "alkylsulfonylamino" group.

[0068] The term "thiol" refers to the -SH group, while "sulfide" refers to the -SR group. 80 The group is included, and "sulfoxide" is -S(O)R 81 The group is included, and "sulfone" is -SO2R 82 The group is included, and "sulfonyl" is -SO2OR 83 The group is included, and "sulfonate" is -SO3 - Includes R 80 , R 81 , R 82 , and R 83 Each of these is independently a substituted or unsubstituted alkyl group, cycloalkyl group, alkenyl group, alkynyl group, aryl group, aralkyl group, heterocyclyl group, or heterocyclylalkyl group as defined herein. In some embodiments, the sulfide is an alkylthio group, i.e., an -S-alkyl group.

[0069] The term "urea" is -NR 84 -C(O)-NR 85 R 86 It refers to the base. R 84 Group, R 85 base, and R 86The group is independently hydrogen, or a substituted or unsubstituted alkyl group, alkenyl group, alkynyl group, cycloalkyl group, aryl group, aralkyl group, heterocyclyl group, or heterocyclylalkyl group as defined herein.

[0070] The term "amidine" is -C(NR 87 )NR 88 R 89 and -NR 87 C(NR 88 )R 89 It refers to R 87 , R 88 , and R 89 Each of these is independently hydrogen, or a substituted or unsubstituted alkyl group, cycloalkyl group, alkenyl group, alkynyl group, aryl group, aralkyl group, heterocyclyl group, or heterocyclylalkyl group as defined herein.

[0071] The term "guanidine" is -NR 90 C(NR 91 )NR 92 R 93 It refers to R 90 , R 91 , R 92 , and R 93 Each of these is independently hydrogen, or a substituted or unsubstituted alkyl group, cycloalkyl group, alkenyl group, alkynyl group, aryl group, aralkyl group, heterocyclyl group, or heterocyclylalkyl group as defined herein.

[0072] The term "enamin" is -C(R 94 )=C(R 95 )NR 96 R 97 and -NR 94 C(R 95 )=C(R 96 )R 97 It refers to R 94 , R 95 , R 96 , and R 97Each of these is independently hydrogen, a substituted or unsubstituted alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group, an aralkyl group, a heterocyclyl group, or a heterocyclylalkyl group as defined herein.

[0073] The terms "halogen" or "halo" as used herein refer to bromine, chlorine, fluorine, or iodine. In some embodiments, halogen is fluorine. In other embodiments, halogen is chlorine or bromine.

[0074] When used herein, the term "hydroxyl" refers to -OH or its ionized form, -O - This can refer to the following. The "hydroxyalkyl" group is a hydroxyl-substituted alkyl group such as HO-CH2-.

[0075] The term "imide" is -C(O)NR 98 C(O)R 99 It refers to R 98 and R 99 Each of these is independently hydrogen, or a substituted or unsubstituted alkyl group, cycloalkyl group, alkenyl group, alkynyl group, aryl group, aralkyl group, heterocyclyl group, or heterocyclylalkyl group as defined herein.

[0076] The term "imin" is -CR 100 (NR 101 ) group and -N(CR 100 R 101 ) refers to the base, R 100 and R 101 Each is independently hydrogen, or a substituted or unsubstituted alkyl group, cycloalkyl group, alkenyl group, alkynyl group, aryl group, aralkyl group, heterocyclyl group, or heterocyclylalkyl group as defined herein, but R 100 and R 101 The condition is that both are not hydrogen at the same time.

[0077] As used herein, the term "nitro" refers to the -NO2 group.

[0078] When used herein, the term "trifluoromethyl" refers to -CF3.

[0079] The term "trifluoromethoxy" as used herein refers to -OCF3.

[0080] The term "Azid" refers to -N3.

[0081] The term "trialkylammonium" refers to a -N(alkyl)3 group. The trialkylammonium group is positively charged and therefore typically has a bonded anion, such as a halogen anion.

[0082] The term "isocyano" refers to -NC.

[0083] The term "isothiocyan" refers to -NCS.

[0084] The term "pentafluorosulfanil" refers to -SF5.

[0085] As will be understood by those skilled in the art, "molecular weight" (also known as "relative molar mass") is a dimensionless quantity, but can be converted to molar mass by multiplying by 1 g / mol or 1 Da. For example, a compound with a weight-average molecular weight of 5,000 has a weight-average molar mass of 5,000 g / mol and a weight-average molar mass of 5,000 Da.

[0086] pharmaceutically acceptable salts of the compounds described herein are within the scope of this technology and include acid-addition salts or base-addition salts that retain the desired pharmacological activity and are not biologically undesirable (e.g., the salts are not excessively toxic, allergenic, or irritating, and are bioavailable). If the compounds of this technology have a basic group, such as an amino group, they can form pharmaceutically acceptable salts with inorganic acids (e.g., hydrochloric acid, hydroboric acid, nitric acid, sulfuric acid, and phosphoric acid), organic acids (e.g., alginates, formic acid, acetic acid, benzoic acid, gluconic acid, fumaric acid, oxalic acid, tartaric acid, lactic acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, and p-toluenesulfonic acid), or acidic amino acids (e.g., aspartic acid and glutamic acid). If the compounds of this technology have an acidic group, such as a carboxylic acid group, they can form pharmaceutically acceptable salts with metals such as alkali metals and alkaline earth metals (e.g., Na + Li + , K + Ca 2+ Mg 2+ Zn 2+ ), ammonia or organic amines (e.g., dicyclohexylamine, trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine), or basic amino acids (e.g., arginine, lysine, and ornithine) can form salts. Such salts can be prepared in situ during the isolation and purification of the compound, or by reacting the purified compound separately with a suitable acid or base in its free base or free acid form, respectively, and then isolating the salt formed.

[0087] Those skilled in the art will recognize that the compounds of this technology may exhibit phenomena of tautomerism, conformational isomerism, geometric isomerism, and / or stereoisomerism. Since the diagrams in this specification and the claims may only represent one of the possible tautomeristic, conformational isomeristic, stereochemical, or geometric isomeristic forms, it should be understood that this technology encompasses any tautomeristic, conformational isomeristic, stereochemical, and / or geometric isomeristic forms of compounds having one or more of the uses described herein, as well as mixtures of various different forms thereof.

[0088] A "tautomer" refers to an isomer of a compound that exists in equilibrium with itself. The presence and concentration of these isomers depend on the environment in which the compound is found, for example, whether the compound is a solid or in an organic solution or aqueous solution. For example, in aqueous solution, quinazolinone may exhibit the following isomers, which are referred to as tautomers of each other: TIFF2026522611000011.tif28128. As another example, guanidine can exhibit the following isomers in protic organic solutions, which are also referred to as tautomers of each other: TIFF2026522611000012.tif29128. Due to the limitations of representing compounds by structural formulas, it will be understood that all chemical formulas of compounds described herein represent all tautomerized forms of the compounds, and that they are within the scope of this art.

[0089] The stereoisomers (also known as optical isomers) of a compound include all chiral, diastereomer, and racemic forms of the structure unless a specific stereochemistry is explicitly stated. Therefore, the compounds used in this technique include optical isomers enriched or divided at any or all chiral atoms, as is evident from the description. Both racemic and diastereomer mixtures, as well as individual optical isomers, can be isolated or synthesized so as to be substantially free of their enantiomer or diastereomer partners, and all such stereoisomers are within the scope of this technique.

[0090] leukemia Leukemia is a type of blood cancer that typically begins in the bone marrow and results in a large number of abnormal blood cells. These blood cells do not fully develop and are called blast cells or leukemia cells. Symptoms may include bleeding and internal bleeding, bone pain, fatigue, fever, and an increased risk of infection. These symptoms result from the absence of normal blood cells. Diagnosis is typically made by blood tests or bone marrow biopsy.

[0091] There are four main types of leukemia: acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), and chronic myeloid leukemia (CML), in addition to several rare types. Both leukemia and lymphoma belong to a broader group of tumors that affect the blood, bone marrow, and lymphatic systems, known as tumors of hematopoietic and lymphoid tissues.

[0092] Mutations in TP53 are present in approximately 10% of patients with AML and myelodysplastic syndromes (MDS) and represent a specific subtype associated with poor outcomes. TP53 is located on chromosome 17, p13, and is essential for cell cycle control and the DNA damage response. TP53 mutations lead to a dominant-negative effect and typically occur in the founder clone that proliferates after cytotoxic stress. Patients with TP53-mutated AML have a very poor prognosis and essentially lack a persistent response to all current treatments. TP53-Y220C is a recurrent hotspot TP53 mutation found in numerous solid tumors and hematological malignancies.

[0093] TP53 reactivated indole derivative TP53 reactivated indole derivatives are known in the art and are described in U.S. Patent No. 10,640,485, the contents of which are incorporated herein by reference in their entirety.

[0094] The TP53 reactivated indole derivative is given by formula (I): It may have the formula TIFF2026522611000013.tif29128, or may be a pharmaceutically acceptable salt thereof. During the ceremony, each TIFF2026522611000014.tif2128 is independently either a single bond or a double bond; X 1 CR 5 , CR 5 R 6 , N, NR 5 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 2 CR 7 , CR 7 R 8 , N, NR 7 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 3 CR 9 , CR 9 R10 , N, NR 9 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 4 CR 11 , CR 11 R 12 , N, NR 11 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 5 CR 13 , N, or NR 13 and; X 1 , X 2 , X 3 , and X 4 At least one of them is Q 1 It is a carbon atom linked to; Q 1 C=O, C=S, C=CR 14 R 15 , C=NR 14 , alkylene, alkenylene, or alkynylene (each of these independently substituted or unsubstituted), bond; m is 1, 2, 3, or 4; Y is either N or O; R 1 is -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 , -R 16 Ure 17 , -R 16 Ure 17 R 18 , -SR 16 , -R 16 SR 17 , -R 16 SR 17 R 18 , -NR 16 R 17 , -R 16 NR 17 , -R 16 NR 17 R 18 , -NR16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 -C(O)R 19 , -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 , -SO2R 19 , alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently unsubstituted or substituted), or hydrogen; Each R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 , halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently unsubstituted or substituted), or hydrogen; Each R 19 and R 20 C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R 21 and R 22 is independently an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R 23 and R 24 These are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen.

[0095] The TP53 reactivated indole derivative is given by formula (IA): It may have the formula TIFF2026522611000015.tif29128, or may be a pharmaceutically acceptable salt thereof. During the ceremony, X 1 CR 5 , NR 5 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 2 CR 7 CH, NR 7 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 3 CR 9CH, NR 9 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 4 CR 11 CH, NR 11 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 5 CR 13 CH, or NR 13 and; X 1 , X 2 , X 3 , and X 4 At least one of them is Q 1 It is a carbon atom linked to; Q 1 It is a combination; m is 1, 2, 3, or 4; Y is either N or O; R 1 is -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 , -R 16 Ure 17 , -R 16 Ure 17 R 18 , -SR 16 , -R 16 SR 17 , -R 16 SR 17 R 18 , -NR 16 R 17 , -R 16 NR 17 , -R 16 NR 17 R 18 , -NR 16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 -C(O)R 19 , -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 , -SO2R 19 , hydrogen, alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl, where each of alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl is independently unsubstituted, or substituted with one or two groups selected from alkyl or halogen, or R 3 and R 4 R 3 and R 4 It forms a ring together with the bonded Y atom, where the ring is either substituted or unsubstituted, or R 3 He is absent; Each R 2 , R 5 , R 7 , R 9 , R 11 , R 13 , R 16 , R 17 , and R 18 -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21, hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, and each alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl is independently unsubstituted or -C(O)R 25 , -C(O)OR 25 , -C(O)NR 25 , -CR 25 3, -OR 25 , -SR 25 , -NR 25 R 26 , -NR 25 C(O)R 26 , -OC(O)R 25 , substituted with one or two groups selected from alkyl, alkenyl, or alkynyl, and R 25 and R 26 are each independently alkyl, hydrogen, or halogen; each R 19 and R 20 are -C(O)R 23 , -C(O)OR 23 , -C(O)NR[[ID=三十七]] 23 R 24 , -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 , -OC(O)R<00009​​​​​​​​​​​​These are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen.

[0096] The TP53 reactivated indole derivative is given by formula (IB): It may have the formula TIFF2026522611000016.tif38128, or may be a pharmaceutically acceptable salt thereof. During the ceremony, X 1 CR 5 , NR 5 , a carbon atom linked with O, S, C=O, C=S, or Y; X 2 CR 7 CH, NR 7 , a carbon atom linked with O, S, C=O, C=S, or Y; X 3 CR 9 CH, NR 9 , a carbon atom linked with O, S, C=O, C=S, or Y; X 4 CR 11 CH, NR 11 , a carbon atom linked with O, S, C=O, C=S, or Y; X 5 CR 13 CH, or NR 13 and; X 1 , X 2 , X 3 , and X 4 At least one of them is a carbon atom linked to Y; Y is either N or O; R 1 is -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 , -R 16 Ure 17 , -R 16 Ure 17 R18 、 -SR 16 、 -R 16 SR 17 、 -R 16 SR 17 R 18 、 -NR 16 R 17 、 -R 16 NR 17 、 -R 16 NR 17 R[[ID=*26]] 18 、 -NR 16 C(O)R 16 、 -OC(O)R 16 、 C=O、 C=S、 -CN、 -SiR 16 R 17 R 18 、 alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which may be independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 is independently, -C(O)R 19 、 -C(O)OR 19 、 -C(O)NR 19 R 20 、 -SOR 19 、 -SO2R 19 、 hydrogen, alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl, where each of alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl is independently unsubstituted or substituted with one or two groups selected from alkyl or halogen, or R 3 and R 4 is, R 3 and R 4 together with the Y atom to which they are attached form a ring, where the ring is substituted or unsubstituted, or R 3 is absent; Each R 2 、 R 5 、 R 7 、 R [[ID=7*6]] 9 、 R 11 、 R 13 Note: There seems to be a potential error in the original text where the numbering in "" and "" might be incorrect as they are not in sequential order as the rest. Also, the translation is done as per the provided rules while trying to maintain the integrity of the original text which appears to be a chemical or patent - related text with some complex notations., R 16 , R 17 , and R 18 -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 ,-CR 25 3, -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 , hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, and each of the alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl is independently unsubstituted or -C(O)R 25 , -C(O)OR 25 -C(O)NR 25 ,-CR 25 3, -OR 25 , -SR 25 , -NR 25 R 26 , -NR 25 C(O)R 26 -OC(O)R 25 It is substituted with one or two groups selected from alkyl, alkenyl, or alkynyl groups, R 25 and R 26 Each of these is independently an alkyl, hydrogen, or halogen; Each R 19 and R 20 C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R 21 and R 22 is independently an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R 23 and R 24 These are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen.

[0097] A non-limiting example of a TP53 reactivated indole derivative is the following formula: One of the following compounds is an example: TIFF2026522611000017.tif21064, TIFF2026522611000018.tif20475, or TIFF2026522611000019.tif134128.

[0098] In some embodiments, the indole derivative is given by formula: It is a compound of TIFF2026522611000020.tif23128, or a pharmaceutically acceptable salt thereof. During the ceremony, each TIFF2026522611000021.tif1128 is independently single-bonded or double-bonded;X 1 CR 5 , CR 5 R 6 , N, NR 5 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to X 2 CR 7 , CR 7 R 8 , N, NR 7 , O, S, C=O, C=S, or Q 1It is a carbon atom linked to X 3 CR 9 , CR 9 R 10 , N, NR 9 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to X 4 CR 11 , CR 11 R 12 , N, NR 11 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to X 5 CR 13 , N, or NR 13 X 1 , X 2 , X 3 , and X 4 At least one of them is Q 1 It is a carbon atom linked to; Q 1 C=O, C=S, C=CR 14 R 15 , C=NR 14 , alkylene, alkenylene, or alkynylene (each of which is independently substituted or unsubstituted), or a bond; m is 1, 2, 3, or 4; Y is N, O, or absent; R 1 is -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 , -SR 16 , -NR 16 R 17 , -NR 16 C(O)R 16 -OC(O)R 16 , -SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently substituted or unsubstituted), or hydrogen; each R 3 and R 4 -C(O)R 19 , -C(O)OR19 -C(O)NR 19 R 20 -SOR 19 , -SO2R 19 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently substituted or unsubstituted), or hydrogen, or R 3 and R 4 R 3 and R 4 It forms a ring together with the nitrogen atom to which it is bonded, and here the ring is either substituted or unsubstituted, or R 3 is absent; each R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which independently substituted or unsubstituted), or hydrogen or halogen; each R 19 and R 20 C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR 23 , -SR 23 , -NR 23 R24 , -NR 23 C(O)R 24 -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which independently substituted or unsubstituted), or hydrogen or halogen; each R 21 and R 22 R is independently an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and each R 23 and R 24 These are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen.

[0099] In some embodiments, the pattern of the dashed bond is selected to yield aromatic systems such as indole, indolene, pyrrolopyridine, pyrrolopyrimidine, or pyrrolopyrazine.

[0100] In some cases, X 1 CR 5 , CR 5 R 6 , or Q 1 It is a carbon atom linked to X. In some embodiments, X 2 CR 7 , CR 7 R 8 , or Q 1 It is a carbon atom linked to X. In some embodiments, X 3 CR 9 , CR 9 R 10 , or Q 1 It is a carbon atom linked to X. In some embodiments, X 4 CR 11 , CR 11 R 12 , or Q 1It is a carbon atom linked to X. In some embodiments, X 5 CR 13 , N, or NR 13 In some cases, X 1 Q 1 It is a carbon atom linked to X. In some embodiments, X 2 Q 1 It is a carbon atom linked to X. In some embodiments, X 3 Q 1 It is a carbon atom linked to X. In some embodiments, X 4 Q 1 It is a carbon atom linked to X. In some embodiments, X 5 It is N.

[0101] In some embodiments, the compound is of the formula: This is from TIFF2026522611000022.tif30128.

[0102] In some embodiments, the compound is of the formula: This is from TIFF2026522611000023.tif94128. In the formula, R 1 is -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 , -SR 16 , -NR 16 R 17 , -NR 16 C(O)R 16 -OC(O)R 16 , SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen.

[0103] In some embodiments, m is 1, 2, 3, or 4. In some embodiments, m is 1. In some embodiments, X 3Q 1 It is a carbon atom linked to a θ, where m is 1. In some embodiments, the compound is given by formula: This is from TIFF2026522611000024.tif73128. In the formula, R 1 is -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 , -SR 16 , -NR 16 R 17 , -NR 16 C(O)R 16 -OC(O)R 16 , SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen.

[0104] In some embodiments, R is alkyl, alkenyl, -C(O)R 16 , -C(O)OR 16 , or -C(O)NR 16 R 17 In some embodiments, R 1 R is a substituted alkyl group. 1 R may be substituted with one or more substituents selected from hydroxyl groups, sulfhydryl groups, halogens, amino groups, nitro groups, nitroso groups, cyano groups, azide groups, sulfoxide groups, sulfone groups, sulfonamide groups, carboxyl groups, carboxyaldehyde groups, imine groups, alkyl groups, haloalkyl groups, cyclic alkyl groups, alkenyl groups, haloalkenyl groups, alkynyl groups, haloalkynyl groups, alkoxy groups, aryl groups, aryloxy groups, aralkyl groups, arylalkoxy groups, heterocyclyl groups, acyl groups, acyloxy groups, carbamate groups, amide groups, urethane groups, and ester groups. In some embodiments, R 1 is an alkyl group substituted with an amine group. In some embodiments, R1 , NR 16 R 17 It is an alkyl group substituted with [a specific compound].

[0105] In some cases, Q 1 C=O, C=S, C=CR 14 R 15 , C=NR 14 , alkylene, alkenylene, or alkynylene (each of which is independently substituted or unsubstituted), or a bond. In some embodiments, Q 1 Q is alkylene, alkenylene, or alkynylene. In some embodiments, Q 1 is C1-alkylene. In some embodiments, each R 16 and R 17 Q is independently alkyl, alkenyl, aryl, heteroaryl, heterocyclyl, or hydrogen. In some embodiments, Q 1 It is a C1-alkylene, and R 16 is an aryl, and R 17 is alkyl. In some embodiments, Q 1 It is a C1-alkylene, and R 16 is an aryl, and R 17 Q is hydrogen. In some embodiments, Q 1 It is a C1-alkylene, and R 16 It is a heteroaryl, and R 17 is alkyl. In some embodiments, Q 1 It is a C1-alkylene, and R 16 It is a heteroaryl, and R 17 Q is hydrogen. In some embodiments, Q 1 It is a C1-alkylene, and R 16 is a substituted heteroaryl, and R 17 Q is hydrogen. In some embodiments, Q 1 It is a C1-alkylene, and R 16 is a substituted alkyl, and R 17 is hydrogen. In some embodiments, R 17R is an aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted with a halogen, alkyl, or hydroxyl. In some embodiments, R 16 is hydrogen, and R 17 R is a halogen- or alkyl-substituted or unsubstituted aryl or heteroaryl. In some embodiments, R 16 is alkyl, and R 17 is a heteroaryl substituted with a halogen or alkyl. In some embodiments, R 17 R is an aryl, heteroaryl, or heterocyclyl, each independently substituted or unsubstituted with an alkyl group. In some embodiments, R 17 is an aryl or heteroaryl group, each of which is independently substituted with an alkyl group, which may be substituted with fluorine, chlorine, bromine, iodine, or cyano.

[0106] In some embodiments, R 2 R is hydrogen or alkyl. In some embodiments, R 13 is an alkyl, alkenyl, hydrogen, or halogen. In some embodiments, R 2 is alkyl, and R 13 is alkyl. In some embodiments, R 2 is hydrogen, and R 13 is alkyl. In some embodiments, R 2 is methyl, ethyl, propyl, isopropyl, butyl, or tert-butyl. In some embodiments, R 13 is methyl, ethyl, propyl, isopropyl, butyl, or tert-butyl. In some embodiments, R 2 is hydrogen, and R 13 It is hydrogen.

[0107] In some embodiments, R 3 is -C(O)R 19 , -C(O)OR 19, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently substituted or unsubstituted), or hydrogen, R 4 is -C(O)R 19 , -C(O)OR 19 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen. In some embodiments, R 3 is hydrogen, and R 4 is a substituted alkyl group. In some embodiments, R 3 is hydrogen, and R 4 is an alkyl group substituted with an aryl group. In some embodiments, R 3 is alkyl, and R 4 is alkyl. In some embodiments, R 3 is alkyl, and R 4 That is Ariel.

[0108] In some embodiments, R 3 is hydrogen, and R 4 It is a heterocycline. In some embodiments, the formula is: This is from TIFF2026522611000025.tif105128.

[0109] In some embodiments, R 3 and R 4 R 3 and R 4 It forms a ring with the bonded nitrogen atom, and the ring is either substituted or unsubstituted. In some embodiments, R 3 and R 4 R 3 and R 4 Together with the bonded nitrogen atom, it forms a substituted heterocycle. In some embodiments, R 3 and R 4 R 3 and R 4Together with the nitrogen atom to which it is bonded, it forms a heterocycle substituted with a hydroxyl group, halogen, amino group, or alkyl group. In some embodiments, R 3 and R 4 R 3 and R 4 It forms a heterocycle with the nitrogen atom to which it is bonded, and the heterocycle is substituted by substituted or unsubstituted heterocycles.

[0110] In some embodiments, R 3 and R 4 R 3 and R 4 Together with the nitrogen atom to which it is bonded, it forms the following equation: It forms a ring in TIFF2026522611000026.tif64128.

[0111] In some embodiments, R 16 is an alkyl, alkenyl, aryl, heteroaryl, heterocyclyl, or hydrogen, and R 17 is an aryl, heteroaryl, or heterocyclyl. In some embodiments, R 17 These are phenyl, indolyl, piperidinyl, imidazolyl, thiazolyl, morpholinyl, pyrrolyl, or pyridinyl.

[0112] In some embodiments, the compound is of the formula: This is from TIFF2026522611000027.tif91128.

[0113] In some embodiments, the compound is of the formula: This is from TIFF2026522611000028.tif81128.

[0114] In some embodiments, the compound is of the formula: This is from TIFF2026522611000029.tif34128. In the formula, each Z 1 and Z 2 CR is independent.x or N; each R x -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently substituted or unsubstituted), hydrogen, or halogen; each R 25 and R 26 -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -SO2R 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 These are alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen.

[0115] In some embodiments, Z 1 is N. In some embodiments, Z 1 and Z 2 is N. In some embodiments, each R 25 and R 26 It is, independently, a halogen. In some embodiments, R 25 teeth, The filename is TIFF2026522611000030.tif20128.

[0116] In some embodiments, R 25 SO2R 21In some embodiments, R 25 SO2R 21 And R 21 is alkyl. In some embodiments, R 25 SO2R 21 And R 21 It is methyl.

[0117] Non-limiting examples of the compounds of this disclosure are listed below: TIFF2026522611000031.tif191128TIFF2026522611000032.tif20872TIFF202 6522611000033.tif196116TIFF2026522611000034.tif44129TIFF2026522611 000035.tif21364TIFF2026522611000036.tif19774TIFF2026522611000037.t if19774TIFF2026522611000038.tif19459TIFF2026522611000039.tif186128 Examples include TIFF2026522611000040.tif20869, TIFF2026522611000041.tif20887, TIFF2026522611000042.tif20387, TIFF2026522611000043.tif107128, TIFF2026522611000044.tif21195, TIFF2026522611000045.tif21578, TIFF2026522611000046.tif21660, TIFF2026522611000047.tif95128, or any pharmaceutically acceptable salt of any of the above.

[0118] Non-limiting examples of the compounds of this disclosure are listed below: TIFF2026522611000048.tif21669TIFF2026522611000049.tif21673TIFF2026522611000050.tif2147 4TIFF2026522611000051.tif20882TIFF2026522611000052.tif193106TIFF2026522611000053.tif20 Examples include 7102TIFF2026522611000054.tif26128TIFF2026522611000055.tif21673TIFF2026522611000056.tif20481TIFF2026522611000057.tif21280TIFF2026522611000058.tif174128, or their pharmaceutically acceptable salts.

[0119] Non-limiting examples of the compounds of this disclosure are listed below: TIFF2026522611000059.tif132128TIFF2026522611000060.tif141128TIFF2026522611000061.tif139128TIFF20265226110 00062.tif144128TIFF2026522611000063.tif157128TIFF2026522611000064.tif142128TIFF2026522611000065.tif127128T Examples include IFF2026522611000066.tif137128, TIFF2026522611000067.tif156155, TIFF2026522611000068.tif155151, TIFF2026522611000069.tif210153, TIFF2026522611000070.tif222137, TIFF2026522611000071.tif117157, or any pharmaceutically acceptable salt of any of the above.

[0120] In some embodiments, the compound is of the formula: It is either TIFF2026522611000072.tif23128 or a pharmaceutically acceptable salt thereof. In the formula, each Q 1a and Q1b These are independently C=O, C=S, and C=CR. 14’ R 15’ , C=NR 14’ , alkylene, alkenylene, or alkynylene (each of these independently substituted or unsubstituted), or bond; each R 1a and R 1b -C(O)R 16’ , -C(O)OR 16’ -C(O)NR 16’ R 7’ , -OR 16’ , -SR 16’ , -NR 16’ R 17’ , -NR 16’ C(O)R 16’ -OC(O)R 16’ , -SiR 16’ R 17’ R 8’ , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently substituted or unsubstituted), or hydrogen; each R 3a and R 3b R is independently alkylene, alkenylene, alkynylene, arylene, heteroarylene, or heterocyclylene (each of which is independently substituted or unsubstituted), or hydrogen; each R 4a and R 4b Independent, absent, -C(O)R 19’ , -C(O)OR 19’ -C(O)NR 19’ R 20’ -SOR 19’ , -SO2R 19’ , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently substituted or unsubstituted), or hydrogen; each R 2a , R 2b , R 13a , and R 13b -C(O)R 21’ , -C(O)OR 21’ -C(O)NR21’ R 22’ , -OR 21’ , -SR 21’ , -NR 21’ R 22’ , -NR 21’ C(O)R 22’ -OC(O)R 21’ , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; each R 19’ and R 20’ -C(O)R 23’ , -C(O)OR 23’ -C(O)NR 23’ R 24’ , -OR 23’ , -SR 23’ , -NR 23’ R 24’ , -NR 23’ C(O)R 24’ -OC(O)R 23’ , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; each R 21’ and R 22’ R is independently an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; each R 23’ and R 24’ L is independently an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; 1 is the linker part; and L 2 This is the linker section.

[0121] In some embodiments, each L 1 and L 2These are independently an ester group, an ether group, a thioether group, a polyethylene glycol (PEG) group, an alkylene group, an alkenylene group, an alkylylene group, a heteroalkylene group, a cycloalkylene group, a heterocyclylene group, an arylene group, a heteroarylene group, or a heterocycloalkylene group, any of which are substituted or unsubstituted. In some embodiments, each L 1 and L 2 These are independently alkylene, alkenylene, alkylylene, heteroalkylene, cycloalkylene, heterocycloalkylene, cycloarylene, or heterocycloalkylene. In some embodiments, L 1 It is alkylene, L 2 It is an ester.

[0122] The compounds described herein may include all of their stereoisomers, enantiomers, diastereomers, mixtures, racemates, atropisomers, and tautomers.

[0123] Non-limiting examples of optional substituents include hydroxyl groups, sulfhydryl groups, halogens, amino groups, nitro groups, nitroso groups, cyano groups, azide groups, sulfoxide groups, sulfone groups, sulfonamide groups, carboxyl groups, carboxyaldehyde groups, imine groups, alkyl groups, haloalkyl groups, alkenyl groups, haloalkenyl groups, alkynyl groups, haloalkynyl groups, alkoxy groups, aryl groups, aryloxy groups, aralkyl groups, arylalkoxy groups, heterocyclyl groups, acyl groups, acyloxy groups, carbamate groups, amide groups, ureido groups, epoxy groups, and ester groups.

[0124] Non-limiting examples of alkyl and alkylene groups include linear, branched, and cyclic alkyl and alkylene groups. Examples of alkyl or alkylene groups include substituted or unsubstituted C1, C2, C3, C4, C5, C6, C7, C8, C9, and C2 alkyl groups. 10 group, C 11 group, C 12 group, C13 group, C 14 group, C 15 group, C 16 group, C 17 group, C 18 group, C 19 group, C 20 group, C 21 group, C 22 group, C 23 group, C 24 group, C 25 group, C 26 group, C 27 group, C 28 group, C 29 group, C 30 group, C 31 group, C 32 group, C 33 group, C 34 group, C 35 group, C 36 group, C 37 group, C 38 group, C 39 group, C 40 group, C 41 group, C 42 group, C 43 group, C 44 group, C 45 group, C 46 group, C 47 group, C 48 group, C 49 base, or C 50 It could be the basis.

[0125] Non-limiting examples of linear alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl.

[0126] Branched alkyl groups include any linear alkyl group substituted with any number of alkyl groups. Non-limiting examples of branched alkyl groups include isopropyl, isobutyl, sec-butyl, and t-butyl.

[0127] Non-limiting examples of substituted alkyl groups include hydroxymethyl, chloromethyl, trifluoromethyl, aminomethyl, 1-chloroethyl, 2-hydroxyethyl, 1,2-difluoroethyl, and 3-carboxypropyl.

[0128] Non-limiting examples of cyclic alkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptlyl, and cyclooctyl groups. Cycloalkyl groups also include condensed bicyclic, crosslinked bicyclic, and spiro-bicyclic structures, as well as higher-order condensed, crosslinked, and spiro systems. Cycloalkyl groups may be substituted with any number of linear, branched, or cyclic alkyl groups. Non-limiting examples of cyclic alkyl groups include cyclopropyl, 2-methyl-cyclopropane-1-yl, cyclopropane-2-en-1-yl, cyclobutyl, 2,3-dihydroxycyclobuta-1-yl, cyclobuta-2-en-1-yl, cyclopentyl, cyclopenta-2-en-1-yl, cyclopenta-2,4-dien-1-yl, cyclohexyl, cyclohexa-2-en-1-yl, cycloheptyl, cyclooctanyl, 2,5-dimethylcyclopenta-1-yl, 3,5-dichlorocyclohexa-1-yl, 4- Examples include hydroxycyclohexa-1-yl, 3,3,5-trimethylcyclohexa-1-yl, octahydropentalenyl, octahydro-1H-indenyl, 3a,4,5,6,7,7a-hexahydro-3H-inden-4-yl, decahydroazlenyl, bicyclo-[2.1.1]hexanyl, bicyclo[2.2.1]heptanyl, bicyclo[3.1.1]heptanyl, 1,3-dimethyl[2.2.1]heptan-2-yl, bicyclo[2.2.2]octanyl, and bicyclo[3.3.3]undecanyl.

[0129] Non-limiting examples of alkenyl and alkenylene groups include linear, branched, and cyclic alkenyl groups. One or more olefins of an alkenyl group may be, for example, E, Z, cis, trans, terminal, or exomethylene. Alkenyl or alkenylene groups may be, for example, substituted or unsubstituted C2, C3, C4, C5, C6, C7, C8, C9, C 10 group, C 11 group, C 12 group, C 13 group, C 14 group, C 15 group, C 16 group, C 17 group, C 18 group, C 19 group, C 20 group, C 21 group, C 22 group, C 23 group, C 24 group, C 25 group, C 26 group, C 27 group, C 28 group, C 29 group, C 30 group, C 31 group, C 32 group, C 33 group, C 34 group, C 35 group, C 36 group, C 37 group, C 38 group, C 39 group, C 40 group, C 41 group, C 42 group, C 43 group, C 44 group, C 45 group, C 46 group, C 47 group, C 48 group, C 49 base, or C 50It may be a group. Non-limiting examples of alkenyl and alkenylene groups include ethenyl, propa-1-en-1-yl, isopropenyl, buta-1-en-4-yl; 2-chloroethenyl, 4-hydroxybuten-1-yl, 7-hydroxy-7-methylocta-4-en-2-yl, and 7-hydroxy-7-methylocta-3,5-dien-2-yl.

[0130] Non-limiting examples of alkynyl or alkynylene groups include linear, branched, and cyclic alkynyl groups. The triple bond in an alkylnyl or alkynylene group can be internal or terminal. Alkylnyl or alkynylene groups include, for example, substituted or unsubstituted C2, C3, C4, C5, C6, C7, C8, C9, C2, C6, C7, C8, C9, C2, C6, C7, C8, C8, C9, C2, C2, C6, C7, C8, C9, C2 10 group, C 11 group, C 12 group, C 13 group, C 14 group, C 15 group, C 16 group, C 17 group, C 18 group, C 19 group, C 20 group, C 21 group, C 22 group, C 23 group, C 24 group, C 25 group, C 26 group, C 27 group, C 28 group, C 29 group, C 30 group, C 31 group, C 32 group, C 33 group, C 34 group, C 35 group, C 36 group, C 37 group, C 38 group, C 39 group, C 40 group, C 41 group, C 42 group, C 43 group, C 44 group, C 45 group, C 46 group, C 47 group, C 48 group, C49 base, or C 50 It may be an alkynyl or alkynylene group. Non-limiting examples of alkynyl or alkynylene groups include ethinyl, propa-2-in-1-yl, propa-1-in-1-yl, and 2-methylhexa-4-in-1-yl; 5-hydroxy-5-methylhexa-3-in-1-yl, 6-hydroxy-6-methylhepta-3-in-2-yl, and 5-hydroxy-5-ethylhepta-3-in-1-yl.

[0131] A haloalkyl group can be any alkyl group substituted with any number of halogen atoms, such as fluorine, chlorine, bromine, and iodine atoms. A haloalkenyl group can be any alkenyl group substituted with any number of halogen atoms. A haloalkynyl group can be any alkynyl group substituted with any number of halogen atoms.

[0132] An alkoxy group can be, for example, an oxygen atom substituted with any alkyl group, alkenyl group, or alkynyl group. Ethers or ether groups include alkoxy groups. Non-limiting examples of alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, and isobutoxy.

[0133] The aryl group may be heterocyclic or nonheterocyclic. The aryl group may be monocyclic or polycyclic. The aryl group may be substituted with any number of substituents described herein, such as hydrocarbyl groups, alkyl groups, alkoxy groups, and halogen atoms. Non-limiting examples of aryl groups include phenyl, toluyl, naphthyl, pyrrolyl, pyridyl, imidazolyl, thiophenyl, and furyl. Non-limiting examples of substituted aryl groups include 3,4-dimethylphenyl, 4-tert-butylphenyl, 4-cyclopropylphenyl, 4-diethylaminophenyl, 4-(trifluoromethyl)phenyl, 4-(difluoromethoxy)phenyl, 4-(trifluoromethoxy)phenyl, 3-chlorophenyl, 4-chlorophenyl, 3,4-dichlorophenyl, 2-fluorophenyl, 2-chlorophenyl, 2-iodophenyl, 3-iodophenyl, 4-iodophenyl, 2-methylphenyl, 3-fluorophenyl, 3-methylphenyl, 3-methoxyphenyl, 4-fluorophenyl, 4-methylphenyl, 4-methoxyphenyl, 2,3-difluorophenyl, 3,4-difluorophenyl, 3,5-difluorophenyl, 2,3-dichlorophenyl, 3,4-dichlorophenyl, 3,5-dichlorophenyl, 2-hydroxyphenyl, 3-hydroxyphenyl, 4-hydroxyphenyl Droxyphenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2,3-dimethoxyphenyl, 3,4-dimethoxyphenyl, 3,5-dimethoxyphenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 2,3,4-trifluorophenyl, 2,3,5-trifluorophenyl, 2,3,6-trifluorophenyl, 2,4,5-trifluorophenyl, 2,4,6-trifluorophenyl, 2,4-dichlorophenyl, 2,5-dichlorophenyl, 2,6-dichlorophenyl, 3,4-dichlorophenyl, 2,3,4-trichlorophenyl, 2,3,5-trichlorophenyl, 2,3,6-trichlorophenyl, 2,4,5-trichlorophenyl, 3,4,5-trichlorophenyl, 2,4,6-trichlorophenyl, 2,3-dimethylphenyl, 2,4-dimethylphenyl, 2,5-dimethylphenyl, 2,Examples include 6-dimethylphenyl, 2,3,4-trimethylphenyl, 2,3,5-trimethylphenyl, 2,3,6-trimethylphenyl, 2,4,5-trimethylphenyl, 2,4,6-trimethylphenyl, 2-ethylphenyl, 3-ethylphenyl, 4-ethylphenyl, 2,3-diethylphenyl, 2,4-diethylphenyl, 2,5-diethylphenyl, 2,6-diethylphenyl, 3,4-diethylphenyl, 2,3,4-triethylphenyl, 2,3,5-triethylphenyl, 2,3,6-triethylphenyl, 2,4,5-triethylphenyl, 2,4,6-triethylphenyl, 2-isopropylphenyl, 3-isopropylphenyl, and 4-isopropylphenyl.

[0134] Non-limiting examples of substituted aryl groups include 2-aminophenyl, 2-(N-methylamino)phenyl, 2-(N,N-dimethylamino)phenyl, 2-(N-ethylamino)phenyl, 2-(N,N-diethylamino)phenyl, 3-aminophenyl, 3-(N-methylamino)phenyl, 3-(N,N-dimethylamino)phenyl, 3-(N-ethylamino)phenyl, 3-(N,N-diethylamino)phenyl, 4-aminophenyl, 4-(N-methylamino)phenyl, 4-(N,N-dimethylamino)phenyl, 4-(N-ethylamino)phenyl, and 4-(N,N-diethylamino)phenyl.

[0135] A heterocycle can be any ring containing a non-carbon ring atom, such as N, O, S, P, Si, B, or any other heteroatom. A heterocycle can be substituted with any number of substituents, such as alkyl groups and halogen atoms. A heterocycle can be aromatic (heteroaryl) or non-aromatic. Non-limiting examples of heterocycles include pyrrole, pyrrolidine, pyridine, piperidine, succinamide, maleimide, morpholine, imidazole, thiophene, furan, tetrahydrofuran, pyran, and tetrahydropyran.

[0136] Non-restrictive examples of heterocycles include diazilinyl, azilidinyl, azetidinyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolinyl, oxathiazolidinol, oxazolidinol, hydantoinyl, tetrahydrofuranyl, pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, dihydropyranyl, tetrahydropyranyl, piperidine-2-onyl, 2,3,4,5-tetrahydro-1H-azepinyl, 2,3-dihydro-1H-indole, and 1,2,3,4-tetrahydro-1H-azepinyl. ii) Heterocyclic units having a single ring containing one or more heteroatoms, with trahydroquinoline being a non-limiting example; and heterocyclic units having two or more rings, one of which is a heterocyclic ring, with hexahydro-1H-pyrrolidinyl, 3a,4,5,6,7,7a-hexahydro-1H-benzo[d]imidazolyl, 3a,4,5,6,7,7a-hexahydro-H-indolyl, 1,2,3,4-tetrahydroquinolinyl, and decahydro-1H-cycloocta[b]pyrrolyl being non-limiting examples.

[0137] Non-limiting examples of heteroaryls include, i) heteroaryl rings containing a single ring, such as 1,2,3,4-tetrazolyl, [1,2,3]triazolyl, [1,2,4]triazolyl, triazinyl, thiazolyl, 1H-imidazolyl, oxazolyl, isoxazolyl, isothiazolyl, furanyl, thiophenyl, pyrimidinyl, 2-phenylpyrimidinyl, pyridinyl, 3-methylpyridinyl, and 4-dimethylaminopyridinyl, as well as ii) Examples of heteroaryl rings include heteroaryl rings containing two or more fused rings, one of which is a heteroaryl ring, such as 7H-prinyl, 9H-prinyl, 6-amino-9H-prinyl, 5H-pyrrolo[3,2-d]pyrimidinyl, 7H-pyrrolo[2,3-d]pyrimidinyl, pyrido[2,3-d]pyrimidinyl, 4,5,6,7-tetrahydro-1-H-indolyl, quinoxalinyl, quinazolinyl, quinolinyl, 8-hydroxyquinolinyl, and isoquinolinyl.

[0138] In some embodiments, the TP53 reactivating indole derivative is PC14586 (also known as Lezatapopt; see https: / / classic.clinicaltrials.gov / ct2 / show / NCT04585750; PCT / US2023 / 067005, filed May 15, 2023; CAS number 2636846-41-6). PC14586 is a first-in-class oral small molecule p53 reactivator selective for the p53 Y220C mutation. PC14586 is designed to bind non-covalently to the fissure created by the Y220C mutation and restore WT p53 function. However, in a Phase 1 clinical trial of PC14586 in locally advanced or metastatic solid tumors with the TP53 Y220C mutation (NCT04585750), only limited activity, including limited cell death, was demonstrated. PC14586 is: It has the structure of TIFF2026522611000073.tif49128.

[0139] MDM2 inhibitors The mouse double microchromosome 2 homolog (MDM2) is a protein encoded by the MDM2 gene in humans and is also known as the E3 ubiquitin-protein ligase MDM2. MDM2 is an important negative regulator of the p53 tumor suppressor. The MDM2 protein functions both as an E3 ubiquitin ligase that recognizes the N-terminal transactivation domain (TAD) of the p53 tumor suppressor and as an inhibitor of p53 transcriptional activation. While MDM2 inhibitors target TP53 WT cells, they may selectively enrich TP53 mutant leukemia cells. MDM2 inhibitors activate p53, which in turn induces MDM2 and p21 levels, leading to cell cycle arrest but not apoptosis.

[0140] MDM2 inhibitors bind to the MDM2 protein, thereby preventing its binding to the transcriptional activation domain of the tumor suppressor protein p53. MDM2 inhibitors can be selective or non-selective small molecules or biomolecules. Examples of MDM2 inhibitors include, but are not limited to, RG7112, RO5045337, Idasanutrin, Nutrin-3a, RG7388, AMG-232, KRT-232, APG-115, BI-907828, CGM097, Cilemadrin, HDM201, Mirademethane, MEL23, MEL24, and DS-3032b.

[0141] MDM2 inhibitors may include MDM2 degrading agents that degrade the MDM2 protein. MDM2 inhibitors may also include proteolytic chimeras (PROTACs), which are heterobifunctional molecules consisting of a targeted ligand tethered to an E3 ubiquitin ligase recruiting ligand to induce selective degradation of MDM2. Examples of MDM2 degradation agents (PROTACs) include pyrooxyindole MDM2 inhibitors tethered to lenalidomide (e.g., MI-1061), nutrin derivatives tethered to lenalidomide analogs (e.g., idasanutrin, nutrin-3a), YX-02-030 (aRG7112 derivative), and MS3227. For further information, please refer to B. Wang et al. Development of selective small molecule MDM2 degraders based on nutlin, Eur. J. Med. Chem. 2019, 176, 476-491; CM Adams et al., Targeted MDM2 degradation reveals a new vulnerability for p53 inactivated triple negative breast cancer, Cancer Discov. 2023, 13(5), 1210-1229; and BK Marcellino, An MDM2 degrader for treatment of acute leukemias, Leukemia This is explained in more detail in 2022, 37, 370-378.

[0142] BCL-2 inhibitor B-cell lymphoma 2 (BCL-2) proteins are a class of proteins that regulate apoptosis. BCL-2 is variably highly expressed in many hematological malignancies, providing protection against cell death induced by oncogenic stress and external stress.

[0143] BCL-2 inhibitors bind to the BCL-2 protein, thereby blocking its protection from apoptosis. BCL-2 inhibitors can be selective or non-selective small molecules or biomolecules. Examples of BCL-2 inhibitors include, but are not limited to, venetoclax, ovatocrax, svatocrax, maritocrax, gossypol, apogossypol, TW-37, UMI-77, and BDA-366.

[0144] XPO-1 inhibitors Exportin 1 (XPO-1) is a eukaryotic protein that mediates the nuclear export of various proteins and RNAs, and is also known as chromosome region maintenance factor 1 (CRM1). XPO-1 mediates NES-dependent protein transport. It transports hundreds of different proteins out of the nucleus. XPO-1 is involved in the nuclear export of ribosomal subunits. XPO-1 is affected in several cancer types and is therefore considered a target for the development of anticancer drugs. Overexpression of XPO-1 plays a role in the development and progression of both solid tumors and hematological malignancies and is associated with a poor prognosis in patients.

[0145] XPO-1 inhibitors bind to the XPO-1 protein, thereby blocking or substantially reducing its nuclear export, which may lead to increased accumulation of tumor suppressor proteins, reduction of oncoproteins, and increased apoptosis. XPO-1 inhibitors can be selective or non-selective small molecules or biomolecules. Examples of XPO-1 inhibitors include, but are not limited to, KPT330 (selinexol), XPOVIO, KPT8602 (ertanexol), KPT335, verginexol, and KPT185.

[0146] Formulations comprising the TP53 reactivated indole derivative, MDM2 inhibitor, BCL-2 inhibitor, and / or XPO-1 inhibitor of this technology The pharmaceutical composition of this technology comprises a TP53 reactivated indole derivative (e.g., PC14586) and one or more additional therapeutic agents. The additional therapeutic agents may include MDM2 inhibitors, BCL-2 inhibitors, and / or XPO-1 inhibitors.

[0147] In some embodiments, the pharmaceutical composition comprises an effective amount of a TP53 reactivated indole derivative (e.g., PC14586) and an effective amount of an MDM2 inhibitor. In some embodiments, the pharmaceutical composition comprises an effective amount of a TP53 reactivated indole derivative (e.g., PC14586) and an effective amount of a BCL-2 inhibitor. In some embodiments, the pharmaceutical composition comprises an effective amount of a TP53 reactivated indole derivative (e.g., PC14586) and an effective amount of an XPO-1 inhibitor. In some embodiments, the pharmaceutical composition comprises an effective amount of a TP53 reactivated indole derivative (e.g., PC14586) as well as effective amounts of an MDM2 inhibitor and a BCL-2 inhibitor. In some embodiments, the pharmaceutical composition comprises an effective amount of a TP53 reactivated indole derivative (e.g., PC14586) as well as effective amounts of an MDM2 inhibitor and a BCL-2 inhibitor. In some embodiments, the pharmaceutical composition comprises an effective amount of a TP53 reactivated indole derivative (e.g., PC14586) as well as effective amounts of an MDM2 inhibitor and a XPO-1 inhibitor. In some embodiments, the pharmaceutical composition comprises an effective amount of a TP53 reactivated indole derivative (e.g., PC14586) and an effective amount of an MDM2 inhibitor, a BCL-2 inhibitor, and an XPO-1 inhibitor. In any embodiment, the pharmaceutical composition may comprise pharmaceutically acceptable pharmaceutical excipients, diluents, or carriers compatible with one or more therapeutic substances in the pharmaceutical composition and method of administration.

[0148] The pharmaceutical compositions of this technology can be manufactured by methods well known in the art, particularly by conventional granulation, mixing, dissolution, encapsulation, freeze-drying, or emulsification processes. The compositions may be produced in various forms, including granules, precipitates, or fine particles, powders including freeze-dried powders, tumble-dried powders, or spray-dried powders, amorphous powders, tablets, capsules, syrups, suppositories, injections, emulsions, elixirs, suspensions, or solutions. The formulations may optionally contain solvents, diluents, and other liquid vehicles, dispersions or suspension aids, surfactants, pH adjusters, isotonic agents, thickeners or emulsifiers, stabilizers and preservatives, solid binders, lubricants, and the like, to suit a particular desired dosage form. In certain embodiments, the compositions disclosed herein are formulated for administration to mammals such as humans.

[0149] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, the liquid dosage form may contain, for example, water, or other solvents, solubilizers, and emulsifiers commonly used in the art, such as water, or ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, cyclodextrin, dimethylformamide, oils (specifically, cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, as well as mixtures thereof. Beyond inert diluents, oral compositions may also contain auxiliary agents such as wetting agents, emulsifiers, and suspending agents, sweeteners, flavoring agents, and fragrances.

[0150] For example, injectable preparations such as sterile aqueous or oily suspensions for injection can be formulated according to known techniques using suitable dispersants or wetting agents and suspending agents. Sterile injectable preparations may also be sterile injectable solutions, suspensions, or emulsions in non-toxic, parenterally acceptable diluents or solvents, such as a solution in 1,3-butanediol. Acceptable vehicles and solvents that can be used include water, U.S. Pharmacopeia Ringer's solution, and isotonic sodium chloride solution. Furthermore, sterile non-volatile oils are conventionally used as solvents or suspension media. For this purpose, any non-irritating non-volatile oil, including synthetic monoglycerides or diglycerides, can be used. In addition, fatty acids such as oleic acid are used in the preparation of injectable preparations. Injectable preparations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating a sterilizing agent into a sterile solid composition in a form that can be dissolved or dispersed in sterile water or other sterile injectable media before use. Compositions formulated for parenteral administration may be administered by bolus injection or timed push, or by continuous infusion.

[0151] To extend the effects of the compounds of this disclosure, it is often desirable to slow down the absorption of the compounds from subcutaneous or intramuscular injection. This can be achieved by using liquid suspensions of crystalline or amorphous materials that have poor water solubility. In this way, the absorption rate of the compound depends on its dissolution rate, which may in turn depend on the crystal size and crystalline form. Alternatively, the absorption delay of parenterally administered compound forms is achieved by dissolving or suspending the compound in an oil vehicle. Injectable depot formulations are made by forming a microcapsule matrix of the compound in a biodegradable polymer such as polylactide-polyglycolide. Depending on the ratio of the compound to the polymer and the properties of the particular polymer used, the rate of compound release can be controlled. Other examples of biodegradable polymers include poly(orthoester) and poly(anhydrous). Depot injection formulations are also prepared by encapsulating the compound in liposomes or microemulsions that are compatible with body tissues.

[0152] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with sodium citrate or dicalcium phosphate, and / or at least one inert pharmaceutically acceptable pharmaceutical additive or carrier, such as a) fillers or bulking agents such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; b) binders such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose, and acacia; c) humectants such as glycerol; d) disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; e) solution flow retarders such as paraffin; f) absorption enhancers such as quaternary ammonium compounds; g) wetting agents such as cetyl alcohol and glycerol monostearate; h) absorbents such as kaolin and bentonite clay; and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also include buffering agents such as phosphates or carbonates.

[0153] Similar types of solid compositions may also be used as fillers in soft and hard-filled gelatin capsules using pharmaceutical additives such as lactose or milk sugar and high molecular weight polyethylene glycol and similar materials. Solid dosage forms of tablets, sugar-coated tablets, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings, controlled-release coatings, and other coatings well known in the field of pharmaceutical formulation. They may optionally contain opacifiers and may be compositions in which they release their active ingredients in the intestinal tract only or in a delayed manner. Examples of embedding compositions that can be used include polymers and waxes.

[0154] The active compound may also be in microencapsulated form together with one or more of the aforementioned pharmaceutical additives. In such solid dosage forms, the active compound may be mixed with at least one inert diluent, such as sucrose, lactose, or starch. Such dosage forms may also include additional substances other than inert diluents, as is common practice, such as tableting lubricants and other tableting aids, such as magnesium stearate and crystalline cellulose. In the case of capsules, tablets, and pills, the dosage form may also include buffers. These may optionally contain opacifiers and may be compositions that release the active ingredient in a delayed manner, either in the intestinal tract or in a specific part thereof. Examples of embedding compositions that can be used include polymers and waxes.

[0155] Treatment method of this technology In one aspect, the present disclosure provides a method for treating wild-type p53 leukemia or TP53-Y220C leukemia in a patient in need, comprising the step of administering to the patient an effective amount of a TP53 reactivated indole derivative and an effective amount of an MDM2 inhibitor, a BCL-2 inhibitor, and / or an XPO-1 inhibitor, wherein the TP53 reactivated indole derivative is of formula (I): Having the formula TIFF2026522611000074.tif29128, or a pharmaceutically acceptable salt thereof, During the ceremony, each TIFF2026522611000075.tif2128 is independently either a single bond or a double bond; X 1 CR 5 , CR 5 R 6 , N, NR 5 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 2 CR 7 , CR 7 R 8 , N, NR 7 , O, S, C=O, C=S, or Q 1It is a carbon atom linked to; X 3 CR 9 , CR 9 R 10 , N, NR 9 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 4 CR 11 , CR 11 R 12 , N, NR 11 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 5 CR 13 , N, or NR 13 and; X 1 , X 2 , X 3 , and X 4 At least one of them is Q 1 It is a carbon atom linked to; Q 1 C=O, C=S, C=CR 14 R 15 , C=NR 14 Alkylene, alkenylene, or alkynylene (each of which is independently substituted or unsubstituted), bond; m is 1, 2, 3, or 4; Y is either N or O; R 1 is -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 , -R 16 Ure 17 , -R 16 Ure 17 R 18 , -SR 16 , -R 16 SR 17 , -R 16 SR 17 R 18 , -NR 16 R 17 , -R16 NR 17 , -R 16 NR 17 R 18 , -NR 16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 -C(O)R 19 , -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 , -SO2R 19 , alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently unsubstituted or substituted), or hydrogen; Each R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently unsubstituted or substituted), or hydrogen; Each R 19 and R 20 C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R 21 and R 22 is independently an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R 23 and R 24 These are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen.

[0156] In one aspect, the present disclosure provides a method for selecting leukemia patients for treatment with a TP53 reactivated indole derivative and an additional therapeutic agent, comprising the steps of detecting the expression of wild-type TP53 or TP53-Y220C mRNA or polypeptide in a biological sample obtained from a leukemia patient, and administering to the leukemia patient an effective amount of a TP53 reactivated indole derivative and an effective amount of at least one additional therapeutic agent selected from MDM2 inhibitors, BCL-2 inhibitors, and XPO-1 inhibitors, wherein the TP53 reactivated indole derivative is of formula (I): Having the formula TIFF2026522611000076.tif29128, or a pharmaceutically acceptable salt thereof, During the ceremony, each TIFF2026522611000077.tif2128 is independently either a single bond or a double bond; X 1 CR 5 , CR 5 R 6 , N, NR 5 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 2 CR 7 , CR 7 R 8 , N, NR 7 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 3 CR 9 , CR 9 R 10 , N, NR 9 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 4 CR 11 , CR 11 R 12 , N, NR 11 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 5 CR 13 , N, or NR 13 and; X 1 , X 2 , X 3 , and X 4 At least one of them is Q 1 It is a carbon atom linked to; Q 1 C=O, C=S, C=CR 14 R 15 , C=NR 14, alkylene, alkenylene, or alkynylene (each of these independently substituted or unsubstituted), bond; m is 1, 2, 3, or 4; Y is either N or O; R 1 is -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 , -R 16 Ure 17 , -R 16 Ure 17 R 18 , -SR 16 , -R 16 SR 17 , -R 16 SR 17 R 18 , -NR 16 R 17 , -R 16 NR 17 , -R 16 NR 17 R 18 , -NR 16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 -C(O)R 19 , -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 , -SO2R 19 , alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently unsubstituted or substituted), or hydrogen; Each R 2, R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 , halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently unsubstituted or substituted), or hydrogen; Each R 19 and R 20 C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R 21 and R 22 is independently an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R23 and R 24 These are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen.

[0157] The biological sample may be whole blood, serum, or plasma.

[0158] Additionally or alternatively, in some embodiments of the methods disclosed herein, the TP53 reactivating indole derivative is PC14586.

[0159] Examples of MDM2 inhibitors include, but are not limited to, RG7112, RO5045337, idasanutrin, nutrin-3a, RG7388, AMG-232, KRT-232, APG-115, BI-907828, CGM097, cilemadrin, HDM201, mirademethan, MEL23, MEL24, and DS-3032b. Examples of BCL-2 inhibitors include, but are not limited to, venetoclax, ovatoclax, subatoclax, maritoclax, gossypol, apogossypol, TW-37, UMI-77, and BDA-366. Examples of XPO-1 inhibitors include, but are not limited to, KPT330 (selinexol), XPOVIO, KPT8602 (ertanexol), KPT335, verginexol, and KPT185.

[0160] In addition or alternatively, in some embodiments of the methods disclosed herein, MDM2 inhibitors, BCL-2 inhibitors, and / or XPO-1 inhibitors are administered sequentially, simultaneously, or separately with TP53 reactivated indole derivatives. In certain embodiments, MDM2 inhibitors, BCL-2 inhibitors, XPO-1 inhibitors, and / or TP53 reactivated indole derivatives are administered orally, intravenously, intramuscularly, intraperitoneally, or subcutaneously.

[0161] In any aspect of the methods disclosed herein, leukemia is acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS).

[0162] Additionally or alternatively, in some embodiments of the methods disclosed herein, mRNA expression levels are detected by real-time quantitative PCR (qPCR), digital PCR (dPCR), reverse transcriptase-PCR (RT-PCR), Northern blotting, microarrays, dot or slot blotting, in-situ hybridization, or fluorescence in-situ hybridization (FISH). Additionally or alternatively, in certain embodiments of the methods disclosed herein, polypeptide expression levels are detected by Western blotting, enzyme-linked immunosorbent assay (ELISA), dot blotting, immunohistochemistry, immunofluorescence, immunoprecipitation, immunoelectrophoresis, or mass spectrometry.

[0163] In some embodiments, the method includes administering an effective amount of a TP53 reactivated indole derivative (e.g., PC14586) and an effective amount of an MDM2 inhibitor to a patient. In certain embodiments, the method includes administering an effective amount of a TP53 reactivated indole derivative (e.g., PC14586) and an effective amount of a BCL-2 inhibitor to a patient. In other embodiments, the method includes administering an effective amount of a TP53 reactivated indole derivative (e.g., PC14586) and an effective amount of an XPO-1 inhibitor to a target. In any embodiment of the methods disclosed herein, the MDM2 inhibitor, BCL-2 inhibitor, XPO-1 inhibitor, and / or the TP53 reactivated indole derivative (e.g., PC14586) may be administered as a single composition or as separate compositions.

[0164] In any aspect of the methods disclosed herein, the patient is unresponsive to at least one prior cancer treatment, such as chemotherapy or immunotherapy. In some aspects, the chemotherapy is trioxide, azacitidine, selvidine, cyclophosphamide, cytarabine, daunorubicin hydrochloride, daurismo, dexamethasone, doxorubicin hydrochloride, enasidenib mesylate, gemtuzumab ozogamicin, gilteritinib fumarate, glassegib maleate, idamycin PFS, idarubicin hydrochloride, idohifa, ivosidenib, midostaurine, This includes one or more of the following: mitoxantrone hydrochloride, Mylotarg, ortasidenib, Onureg, Pemazil, pemigatinib, prednisone, Lezlidia, Rituxan, Rituximab, rubidomycin, Lydapt, Tabloid, thioguanine, Tibsovo, tisagenlecleucel, Trisenox, Vencrexta, Venetoclas, vinicristine sulfate, Bixeos, or Zospata. The patient may be a child or an adult. In certain embodiments, the patient is human. Additionally or alternatively, in any embodiment of the methods disclosed herein, the patient includes an acquired TP53-Y220C mutation.

[0165] Additionally or alternatively, some embodiments of the combination therapy methods disclosed herein improve the time to response and / or duration of response compared to monotherapy with a TP53 reactivated indole derivative (e.g., PC14586) or monotherapy with an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor.

[0166] Additionally or alternatively, in some embodiments of the methods disclosed herein, the TP53 reactivated indole derivative (e.g., PC14586) and the MDM2 inhibitor are administered sequentially, simultaneously, or separately. The TP53 reactivated indole derivative (e.g., PC14586) and / or the MDM2 inhibitor may be administered orally, parenterally, by inhalation spray, intranasally, orally, or via an implanted reservoir. The term "parenteral," as used herein, includes injection or infusion techniques into the subcutaneous, intravenous, intramuscular, intra-articular, intra-articular bursa, intrasternal, intrathecal, intrahepatic, intrafocal, and intracranial regions. In some embodiments, the composition is administered orally, intravenously, or subcutaneously. Formulations comprising the TP53 reactivated indole derivative (e.g., PC14586) and / or the MDM2 inhibitor disclosed herein may be designed to be short-acting, immediate-release, or long-acting. In other embodiments, the compound may be administered by local means rather than systemic means, such as administration at the tumor site (e.g., by injection).

[0167] Additionally or alternatively, in some embodiments of the methods disclosed herein, the TP53 reactivated indole derivative (e.g., PC14586) and the BCL-2 inhibitor are administered sequentially, simultaneously, or separately. The TP53 reactivated indole derivative (e.g., PC14586) and / or the BCL-2 inhibitor may be administered orally, parenterally, by inhalation spray, intranasally, orally, or via an implantable reservoir. The term "parenteral," as used herein, includes injection or infusion techniques into the subcutaneous, intravenous, intramuscular, intra-articular, intra-articular bursa, intrasternal, intrathecal, intrahepatic, intrafocal, and intracranial regions. In some embodiments, the composition is administered orally, intravenously, or subcutaneously. Formulations comprising the TP53 reactivated indole derivative (e.g., PC14586) and / or the BCL-2 inhibitor disclosed herein may be designed to be short-acting, immediate-release, or long-acting. In other embodiments, the compound may be administered by local means rather than systemic means, such as administration at the tumor site (e.g., by injection).

[0168] Additionally or alternatively, in some embodiments of the methods disclosed herein, the TP53 reactivated indole derivative (e.g., PC14586) and the XPO-1 inhibitor are administered sequentially, simultaneously, or separately. The TP53 reactivated indole derivative (e.g., PC14586) and / or the XPO-1 inhibitor may be administered orally, parenterally, by inhalation spray, intranasally, orally, or via an implantable reservoir. The term "parenteral," as used herein, includes injection or infusion techniques into the subcutaneous, intravenous, intramuscular, intra-articular, intra-articular bursa, intrasternal, intrathecal, intrahepatic, intrafocal, and intracranial regions. In some embodiments, the composition is administered orally, intravenously, or subcutaneously. Formulations comprising the TP53 reactivated indole derivative (e.g., PC14586) and / or the XPO-1 inhibitor disclosed herein may be designed to be short-acting, immediate-release, or long-acting. In other embodiments, the compound may be administered by local means rather than systemic means, such as administration at the tumor site (e.g., by injection).

[0169] Additionally or alternatively, in some embodiments of the methods disclosed herein, a TP53 reactivated indole derivative (e.g., PC14586) is administered to a patient with leukemia prior to the administration of an MDM2 inhibitor, BCL-2 inhibitor, or XPO-1 inhibitor (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 It can be administered one week prior, two weeks prior, three weeks prior, four weeks prior, five weeks prior, six weeks prior, eight weeks prior, or twelve weeks prior), simultaneously with or afterward (for example, 5 minutes later, 15 minutes later, 30 minutes later, 45 minutes later, 1 hour later, 2 hours later, 4 hours later, 6 hours later, 12 hours later, 24 hours later, 48 hours later, 72 hours later, 96 hours later, 1 week later, 2 weeks later, 3 weeks later, 4 weeks later, 5 weeks later, 6 weeks later, 8 weeks later, or twelve weeks later).

[0170] In some embodiments, a TP53 reactivated indole derivative (e.g., PC14586) and at least one of an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor are administered to a patient, such as a mammal such as a human, in an order and time interval such that the first-administered inhibitor acts together with the second-administered inhibitor to provide a greater benefit than if each inhibitor were administered alone. For example, the TP53 reactivated indole derivative (e.g., PC14586) and at least one of an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor may be administered at the same time or sequentially in any order at different time points, but if not administered at the same time, the TP53 reactivated indole derivative (e.g., PC14586) and at least one of an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor are administered with sufficient time intervals to obtain the desired therapeutic or prophylactic effect of the combination of at least two inhibitors. In one embodiment, a TP53 reactivated indole derivative (e.g., PC14586) and at least one of an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor exert their effects over overlapping time. In some embodiments, a TP53 reactivated indole derivative (e.g., PC14586) and at least one of an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor are each administered as separate dosage forms in any suitable form by any preferred route. In other embodiments, a TP53 reactivated indole derivative (e.g., PC14586) and at least one of an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor are administered simultaneously in a single dosage form.

[0171] It will be recognized that the frequency at which any of these therapeutic substances can be administered may be once or more than once over periods of approximately 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 20 days, 28 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, every 2 months, every 3 months, every 4 months, every 5 months, every 6 months, every 7 months, every 8 months, every 9 months, every 10 months, every 11 months, every 1 year, every 2 years, every 3 years, every 4 years, or every 5 years.

[0172] For example, TP53 reactivated indole derivatives (e.g., PC14586), MDM2 inhibitors, BCL-2 inhibitors, or XPO-1 inhibitors may be administered daily, weekly, bi-weekly, or monthly over a specific period. TP53 reactivated indole derivatives (e.g., PC14586), MDM2 inhibitors, BCL-2 inhibitors, or XPO-1 inhibitors may be taken daily over a 14-day period or twice daily over a 7-day period. ATP53 reactivated indole derivatives (e.g., PC14586), MDM2 inhibitors, BCL-2 inhibitors, or XPO-1 inhibitors may be administered daily over a 7-day period.

[0173] Alternatively, a TP53 reactivated indole derivative (e.g., PC14586), MDM2 inhibitor, BCL-2 inhibitor, or XPO-1 inhibitor may be administered daily, weekly, bi-weekly, or monthly for a specific period, followed by a period of no treatment. In some embodiments, a TP53 reactivated indole derivative (e.g., PC14586), MDM2 inhibitor, BCL-2 inhibitor, or XPO-1 inhibitor may be administered daily for 14 days, followed by 7 days of no treatment, and this cycle of 14 days of daily administration followed by 7 days of no treatment may be repeated more than twice. In some embodiments, a TP53 reactivated indole derivative (e.g., PC14586), MDM2 inhibitor, BCL-2 inhibitor, or XPO-1 inhibitor may be administered twice daily for 7 days, followed by 14 days of no treatment, and this cycle of 7 days of twice daily administration followed by 14 days of no treatment may be repeated one or more times.

[0174] In some embodiments, a TP53 reactivated indole derivative (e.g., PC14586), an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor is administered daily over a period of 14 days. In other embodiments, a TP53 reactivated indole derivative (e.g., PC14586), an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor is administered daily over a period of 12, 11, 10, 9, or 8 days. In other embodiments, a TP53 reactivated indole derivative (e.g., PC14586), an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor is administered daily over a period of 7 days. In other embodiments, a TP53 reactivated indole derivative (e.g., PC14586), an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor is administered daily over a period of 6, 5, 4, or 3 days.

[0175] In some embodiments, individual doses of a TP53 reactivated indole derivative (e.g., PC14586) and an MDM2 inhibitor, BCL-2 inhibitor, or XPO-1 inhibitor are administered within a time interval (e.g., within 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 5 days, 6 days, 1 week, or 2 weeks) that allows the two inhibitors to work together. In some embodiments, a specific duration of no-treatment period follows a treatment period in which the therapeutic substance is administered, during which the therapeutic substance is not administered to the patient. This no-treatment period may be followed by a series of subsequent treatment periods and no-treatment periods of the same or different durations and frequencies. In some embodiments, treatment periods and no-treatment periods are alternated. It will be understood that the treatment period in cycling therapy may continue until the patient achieves a complete or partial response, at which point treatment may be stopped. Alternatively, the duration of treatment in cycling therapy may be continued until the patient achieves a complete or partial response, at which point the duration of treatment may be continued for a specific number of cycles. In some embodiments, the length of the duration of treatment may be a specific number of cycles, regardless of the patient's response. In some other embodiments, the length of the duration of treatment may be continued until the patient relapses.

[0176] In some embodiments, TP53 reactivated indole derivatives (e.g., PC14586) and MDM2 inhibitors, BCL-2 inhibitors, or XPO-1 inhibitors are administered to the patient periodically. Cycling therapy involves administering a first activator (e.g., a first prophylactic or therapeutic agent) over a period of time, followed by administering a second activator and / or a third activator (e.g., a second and / or third prophylactic or therapeutic agent) over a period of time, and repeating this sequence of administrations. Cycling therapy can reduce the development of resistance to one or more of the treatments, avoid or reduce the side effects of one of the treatments, and / or improve the efficacy of the treatment.

[0177] In some embodiments, a TP53 reactivated indole derivative (e.g., PC14586) combined with an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor is administered at doses and schedules typically used for that activator during monotherapy. In other embodiments, a TP53 reactivated indole derivative (e.g., PC14586) and one of the MDM2 inhibitor, BCL-2 inhibitor, or XPO-1 inhibitor are administered concurrently, allowing for advantageous administration of one or both activators at lower doses than those typically administered when the activators are used during monotherapy, resulting in doses below the threshold for inducing adverse side effects.

[0178] The therapeutically effective or preferred dose of a combined TP53 reactivated indole derivative (e.g., PC14586), MDM2 inhibitor, BCL-2 inhibitor, and XPO-1 inhibitor depends on several factors, including the nature of the severity of the condition to be treated, the specific inhibitor, the route of administration, and the individual patient's age, weight, overall health, and response. In certain embodiments, the preferred dose level is one that achieves a therapeutic response as measured by a reduction in cancer cells, or by disease progression, progression-free survival, or other standard measures of overall survival. In other embodiments, the preferred dose level is one that, in addition to achieving this therapeutic response, minimizes any side effects associated with the administration of the therapeutic substance.

[0179] The preferred daily dose of an MDM2 inhibitor may generally range from about 10% to about 120% of the maximum tolerated dose as a monotherapy, in single doses, divided doses, or multiple doses. In certain embodiments, the preferred dose of an MDM2 inhibitor is about 20% to about 100% of the maximum tolerated dose as a monotherapy. In other embodiments, the preferred dose of an MDM2 inhibitor is about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 105%, about 110%, about 115%, or about 120% of the maximum tolerated dose as a monotherapy.

[0180] The preferred daily dose of a BCL-2 inhibitor may generally range from about 10% to about 120% of the maximum tolerated dose as a monotherapy, in single doses, divided doses, or multiple doses. In certain embodiments, the preferred dose of a BCL-2 inhibitor is about 20% to about 100% of the maximum tolerated dose as a monotherapy. In other embodiments, the preferred dose of a BCL-2 inhibitor is about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 105%, about 110%, about 115%, or about 120% of the maximum tolerated dose as a monotherapy.

[0181] The preferred daily dose of an XPO-1 inhibitor may generally range from about 10% to about 120% of the maximum tolerated dose as a monotherapy, in single doses, divided doses, or multiple doses. In certain embodiments, the preferred dose of an XPO-1 inhibitor is about 20% to about 100% of the maximum tolerated dose as a monotherapy. In other embodiments, the preferred dose of an XPO-1 inhibitor is about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 105%, about 110%, about 115%, or about 120% of the maximum tolerated dose as a monotherapy.

[0182] The preferred daily dose of a TP53 reactivated indole derivative (e.g., PC14586) may generally range from about 10% to about 120% of the maximum tolerated dose as a monotherapy, whether in single, divided, or multiple doses. In certain embodiments, the preferred dose of a TP53 reactivated indole derivative (e.g., PC14586) may range from about 20% to about 100% of the maximum tolerated dose as a monotherapy. In other embodiments, preferred drug doses of TP53 reactivated indole derivatives (e.g., PC14586) are about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 105%, about 110%, about 115%, or about 120% of the maximum tolerated dose as a monotherapy.

[0183] The drug dose, toxicity, and therapeutic efficacy of any therapeutic substance can be determined, for example, by standard pharmaceutical procedures in cell culture or experimental animals to determine the LD50 (the dose at which 50% of the population dies) and ED50 (the dose at which 50% of the population is therapeutically effective). The dose-to-toxicity ratio is the therapeutic index and can be expressed as the LD50 / ED50 ratio. Compounds exhibiting a high therapeutic index are advantageous. While compounds exhibiting toxic side effects may be used, care should be taken to design delivery systems that target such compounds to the site of the affected tissue, minimizing potential damage to uninfected cells and thereby reducing side effects.

[0184] Data obtained from cell culture assays and animal studies can be used to determine the range of drug doses for use in humans. The drug dose of such compounds may be within the range of circulating concentrations, including the ED50, which is nearly or completely toxic. Within this range, the drug dose may vary depending on the dosage form used and the route of administration utilized. For any compound used in the method, the therapeutically effective dose can first be estimated from cell culture assays. Doses can be determined in animal models to achieve the circulating plasma concentration range, including the IC50 (i.e., the concentration of the test compound that achieves half of the maximum symptom inhibition) determined in cell culture. Using such information, a useful dose in humans can be accurately determined. Plasma levels can be measured, for example, by high-performance liquid chromatography.

[0185] Typically, the effective dose of a TP53 reactivated indole derivative (e.g., PC14586), MDM2 inhibitor, BCL-2 inhibitor, or XPO-1 inhibitor sufficient to achieve a therapeutic or prophylactic effect may be in the range of about 0.000001 mg / kg body weight / day to about 10,000 mg / kg body weight / day. Preferably, the drug dose range is about 0.0001 mg / kg body weight / day to about 100 mg / kg body weight / day. For example, the drug dose may be 1 mg / kg body weight or 10 mg / kg body weight every day, every two days, or every three days, or in the range of 1 to 10 mg / kg every week, every two weeks, or every three weeks. In one embodiment, the single drug dose of a TP53 reactivated indole derivative (e.g., PC14586), MDM2 inhibitor, BCL-2 inhibitor, or XPO-1 inhibitor is in the range of 0.001 to 10,000 micrograms / kg body weight. In one embodiment, the concentration of the TP53 reactivated indole derivative (e.g., PC14586), MDM2 inhibitor, BCL-2 inhibitor, or XPO-1 inhibitor in the carrier is in the range of 0.2 to 2000 micrograms / delivery milliliter. Exemplary treatment regimens involve administration once daily or once weekly. For therapeutic use, relatively high doses at relatively short intervals may be required until disease exacerbations are reduced or terminated, or until the subject shows partial or complete remission of disease symptoms. Thereafter, a prophylactic regimen may be administered to the patient.

[0186] In some embodiments, the therapeutically effective dose of a TP53 reactivated indole derivative (e.g., PC14586), an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor is defined as a concentration of 10 in the target tissue of the TP53 reactivated indole derivative (e.g., PC14586), an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor. -12 ~10 -6 Molar concentration, for example, approximately 10 -7This can be defined as a molar concentration. This concentration can be delivered by a systemic dose of 0.001–100 mg / kg or an equivalent dose based on body surface area. The dose schedule will be optimized to maintain a therapeutic concentration in the target tissue, including not only single doses daily or weekly, but also continuous infusion (e.g., parenteral infusion or transdermal application).

[0187] Those skilled in the art will recognize that certain factors, including but not limited to the severity of the disease or disorder, prior treatment, the subject's overall health and / or age, and other pre-existing conditions, may influence the dosage and timing of treatment for a subject effectively. Furthermore, treatment of a subject with a therapeutically effective amount of the therapeutic composition described herein may include a single treatment or a series of treatments.

[0188] The mammals treated according to this method may be any mammal, including, for example, livestock such as sheep, pigs, cows, and horses; pet animals such as dogs and cats; and laboratory animals such as rats, mice, and rabbits. In some embodiments, the mammal is a human.

[0189] This technology kit This disclosure provides a kit for treating leukemia, comprising (a) a TP53 reactivating indole derivative (e.g., PC14586), (b) at least one of an MDM2 inhibitor, a BCL-2 inhibitor, and an XPO-1 inhibitor, and (c) instructions for treating the leukemia. In some embodiments, the leukemia is AML and / or MDS. Additionally or alternatively, in some embodiments, the leukemia comprises wild-type p53 or a TP53-Y220C mutation.

[0190] When co-administration is intended, the kit may comprise a TP53 reactivated indole derivative (e.g., PC14586), and at least one of the MDM2 inhibitor, BCL-2 inhibitor, and XPO-1 inhibitor, either formulated as a single pharmaceutical composition such as a tablet, or as separate pharmaceutical compositions. When the TP53 reactivated indole derivative (e.g., PC14586), and one or more of the MDM2 inhibitor, BCL-2 inhibitor, and XPO-1 inhibitors disclosed herein are not administered co-administered, the kit may comprise (a) a TP53 reactivated indole derivative (e.g., PC14586), and (b) at least one of the MDM2 inhibitor, BCL-2 inhibitor, and XPO-1 inhibitor, either formulated as separate pharmaceutical compositions in a single package or separate packages.

[0191] Additionally or alternatively, in some embodiments, the kit further comprises at least one chemotherapeutic agent and / or at least one immunotherapeutic agent useful for treating leukemia. Examples of such chemotherapeutic agents include arsenic trioxide, azacitidine, selvidine, cyclophosphamide, cytarabine, daunorubicin hydrochloride, daurismo, dexamethasone, doxorubicin hydrochloride, enasidenib mesylate, gemtuzumab ozogamicin, gilteritinib fumarate, glassegib maleate, idamycin PFS, idarubicin hydrochloride, idohifa, ivosidenib, midos Examples include taurine, mitoxantrone hydrochloride, Mylotarg, ortasidenib, Onureg, Pemazil, pemigatinib, prednisone, Lezlidia, Rituxan, rituximab, rubidomycin, Lydapt, Tabloid, thioguanine, Tibsovo, tisagenlecleucel, Trisenox, Vencrexta, Venetoclas, vinicristine sulfate, Bixeos, and Zospata. The kit may further comprise pharmaceutically acceptable pharmaceutical excipients, diluents, or carriers compatible with one or more kit components described herein. Optionally, the above components of the kit of this technology are packaged in a suitable container and labeled for the treatment of leukemia, including AML and / or MDS. The kit may optionally include instructions that are customarily included in the commercial packaging of therapeutic products, such instructions may include, for example, information regarding signs, usage, dosage, manufacture, administration, contraindications, and / or warnings relating to the use of such therapeutic products. [Examples]

[0192] This technology is further illustrated by the following embodiments, but these should not be interpreted as limiting it in any way.

[0193] Example 1: Materials and Method PC14586 is supplied by PMV Pharmaceuticals. For further details, see Dumbrava EE, Johnson ML, Tolcher AW, Shapiro G, Thompson JA, El-Khoueiry AB, et al. “First-in-human study of PC14586, a small molecule structural corrector of Y220C mutant p53, in patients with advanced solid tumors harboring a TP53 Y220C mutation.” J Clin Oncol (2022) 16_suppl:3003. doi: 10.1200 / JCO.2022.40.16_suppl.3003.

[0194] The BCL-2 inhibitor was venetoclax (BCL201). The MDM2 inhibitor was nutrin-3a (HDM201). The XPO1 inhibitors were KPT330 and KPT8602.

[0195] In vitro studies were conducted using AML cells with wild-type TP53, as well as isogenic AML cells modified to have TP53 knockout (KO), isogenic AML cells modified to have the TP53-R175H mutation, and isogenic AML cells modified to have the TP53-Y220C mutation.

[0196] Cell culture: AML cells were cultured in RPMI-1640 medium supplemented with 10% inactivated fetal bovine serum, 2 mM L-glutamine, 100 U / mL penicillin, and 100 μg / mL streptomycin. The cells were maintained at 37°C in a humidified 5% CO2 atmosphere.

[0197] Cell viability test:Apoptosis was evaluated by flow cytometry of cells stained with annexin V (AnnV). Cell membrane integrity was simultaneously assessed by elimination of 7-aminoactinomycin D (7AAD) in annexin V-stained cells. The number of viable cells was determined by flow cytometry using counting beads and expressed as a percentage of the control.

[0198] Example 2: PC14586 converts p53Y220C to the wild-type p53 conformation, activates p53 transcriptional activity, and greatly induces p21. Figure 1 shows Western blots (upper panel) demonstrating the activity of PC14586 against mutant and wild-type p53, and Western blots (lower panel) demonstrating the transcriptional activity of p53 after treatment with PC14586. The Western blots clearly show that PC14586 activates p53 and p53 transcriptional activity and induces p21 expression.

[0199] Example 3: PC14586 primarily inhibits cell proliferation in TP53 Y220C AML cells, but does not inhibit cell proliferation in TP53 WT AML cells, TP53 KO AML cells, or TP53 R175H mutant AML cells. Treatment with PC14586 was tested on a group of AML cell lines. The group included AML cell lines containing TP53-WT, TP53-KO, TP53-R175H, or TP53-Y220C. Figure 2A shows the dose-response survival curves for a group of AML cell lines treated with PC14586 for 120 hours. Figure 2B shows the 7AAD / AnnV curves for a group of AML cell lines treated with PC14586 for 120 hours. The survival curves show the percentage of viable cells in the sample, while the 7AAD / AnnV curves reveal the percentage of apoptotic cells in the sample.

[0200] The results from these tests revealed that PC14586 primarily suppresses the proliferation of AML cells with the TP53-Y220C p53 mutation. On the other hand, PC14586 did not substantially affect the percentage of apoptotic cells in any of the samples, including AML cells with the TP53-Y220C p53 mutation, indicating that PC14586 monotherapy does not induce apoptosis in these cell lines.

[0201] Example 4: MDM2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. The combination of an MDM2 inhibitor and PC14586 (MDM2i + PC14586) was tested in vitro and compared to treatment with the MDM2 inhibitor (MDM2i) or PC14586 alone. AML cell lines containing TP53-WT, TP53-KO, TP53-R175H, or TP53-Y220C were treated with the combination of MDM2 and PC14586 for 72 hours. The concentration of MDM2 was varied between 0 μM and 5.00 μM, and the concentration of PC14586 was varied between 0 μM and 4.0 μM.

[0202] Figure 3A shows the 7AAD / AnnV curves of AML cells with TP53-WT treated for 72 hours with an MDM2 inhibitor, PC14586, or a combination of the MDM2 inhibitor and PC14586. Figure 3B shows the dose-response survival curves of AML cells with TP53-WT treated for 72 hours with an MDM2 inhibitor, PC14586, or a combination of the MDM2 inhibitor and PC14586. The results clearly showed that the MDM2 inhibitor enhanced PC14586 activity in AML cells with TP53-WT, resulting in a significant increase in cell death and a decrease in cell proliferation compared to monotherapy.

[0203] Figure 3C shows the 7AAD / AnnV curves for AML cells with TP53-Y220C treated for 72 hours with an MDM2 inhibitor, PC14586, or a combination of the MDM2 inhibitor and PC14586. Figure 3D shows the dose-response survival curves for AML cells with TP53-Y220C treated for 72 hours with an MDM2 inhibitor, PC14586, or a combination of the MDM2 inhibitor and PC14586. The results revealed that the MDM2 inhibitor enhances PC14586 activity in AML cells with TP53-Y220C, and that the combination treatment has a synergistic effect, resulting in a significant increase in cell death and a decrease in cell proliferation compared to monotherapy.

[0204] The results clearly showed that neither the MDM2 inhibitor nor PC14586, nor any combination thereof, had substantial activity in TP53 KO AML cells or TP53 R175H AML cells. Figure 3E is a graph of the 7AAD / AnnV curve for TP53-KO AML cells treated for 72 hours with the MDM2 inhibitor, PC14586, or a combination of the MDM2 inhibitor and PC14586. Figure 3F is a graph of the dose-response survival curve for TP53-KO AML cells treated for 72 hours with the MDM2 inhibitor, PC14586, or a combination of the MDM2 inhibitor and PC14586. Figure 3G is a graph of the 7AAD / AnnV curve for TP53-R175H AML cells treated for 72 hours with the MDM2 inhibitor, PC14586, or a combination of the MDM2 inhibitor and PC14586. Figure 3H is a graph of dose-response survival curves for AML cells with TP53-R175H treated for 72 hours with an MDM2 inhibitor, PC14586, or a combination of an MDM2 inhibitor and PC14586.

[0205] Example 5: BCL-2 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. The combination of BCL-2 inhibitors and PC14586 (BCL-2i + PC14586) was tested in vitro and compared to treatment with BCL-2 inhibitors (BCL-2i) or PC14586 alone. AML cell lines containing TP53-WT, TP53-KO, TP53-R175H, or TP53-Y220C were treated with the combination of BCL-2 and PC14586 for 72 hours. The concentration of BCL-2 was varied between 0 nM and 20.00 nM, and the concentration of PC14586 was varied between 0 μM and 4.0 μM.

[0206] Figure 4A shows the 7AAD / AnnV curves for AML cells with TP53-WT treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. Figure 4B shows the dose-response survival curves for AML cells with TP53-WT treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. The results clearly show that combination therapy has a synergistic effect on AML cells with TP53-WT, resulting in increased cell death and reduced cell proliferation compared to monotherapy.

[0207] Figure 4C shows the 7AAD / AnnV curves for AML cells with TP53-Y220C treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. Figure 4D shows the dose-response survival curves for AML cells with TP53-Y220C treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. The results clearly show that combination therapy has a synergistic effect on AML cells with TP53-Y220C, resulting in increased cell death and reduced cell proliferation compared to monotherapy.

[0208] The combination of a BCL-2 inhibitor and PC14586 slightly increased the activity against TP53 KO AML cells and TP53 R175H AML cells. Figure 4E is a graph of the 7AAD / AnnV curves for AML cells with TP53-KO treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. Figure 4F is a graph of the dose-response survival curves for AML cells with TP53-KO treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. Figure 4G is a graph of the 7AAD / AnnV curves for AML cells with TP53-R175H treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586. Figure 4H is a graph of dose-response survival curves for AML cells with TP53-R175H treated for 72 hours with a BCL-2 inhibitor, PC14586, or a combination of a BCL-2 inhibitor and PC14586.

[0209] Example 6: XPO-1 inhibition enhances PC14586 activity in AML cells with TP53-WT, and this combination is highly synergistic in AML cells with TP53-Y220C. The combination of XPO-1 inhibitors and PC14586 (XPO-1i + PC14586) was tested in vitro and compared to treatment with XPO-1 inhibitor (XPO-1i) or PC14586 alone. AML cell lines containing TP53-WT, TP53-KO, TP53-R175H, or TP53-Y220C were treated with the XPO-1 and PC14586 combination for 72 hours. The concentration of XPO-1 was varied between 0 nM and 200.00 nM, and the concentration of PC14586 was varied between 0 μM and 4.0 μM.

[0210] Figure 5A shows the 7AAD / AnnV curves of AML cells with TP53-WT treated for 72 hours with an XPO inhibitor, PC14586, or a combination of an XPO inhibitor and PC14586. Figure 5B shows the dose-response survival curves of AML cells with TP53-WT treated for 72 hours with an XPO inhibitor, PC14586, or a combination of an XPO inhibitor and PC14586. The results clearly show that combination therapy has a synergistic effect on AML cells with TP53-WT, resulting in increased cell death and reduced cell proliferation compared to monotherapy.

[0211] Figure 5C shows the 7AAD / AnnV curves for AML cells with TP53-Y220C treated for 72 hours with an XPO inhibitor, PC14586, or a combination of an XPO inhibitor and PC14586. Figure 5D shows the dose-response survival curves for AML cells with TP53-Y220C treated for 72 hours with an XPO inhibitor, PC14586, or a combination of an XPO inhibitor and PC14586. The results clearly show that combination therapy has a synergistic effect on AML cells with TP53-Y220C, resulting in increased cell death and reduced cell proliferation compared to monotherapy.

[0212] The combination of the XPO-1 inhibitor and PC14586 slightly increased the activity against TP53 KO AML cells and TP53 R175H AML cells. Figure 5E is a graph of the 7AAD / AnnV curves for TP53-KO AML cells treated for 72 hours with the XPO inhibitor, PC14586, or a combination of the XPO inhibitor and PC14586. Figure 5F is a graph of the dose-response survival curves for TP53-KO AML cells treated for 72 hours with the XPO inhibitor, PC14586, or a combination of the XPO inhibitor and PC14586. Figure 5G is a graph of the 7AAD / AnnV curves for TP53-R175H AML cells treated for 72 hours with the XPO inhibitor, PC14586, or a combination of the XPO inhibitor and PC14586. Figure 5H shows the dose-response survival curves of AML cells with TP53-R175H treated for 72 hours with an XPO inhibitor, PC14586, or a combination of an XPO inhibitor and PC14586.

[0213] Example 7. Synergistic effect of combination therapy of this technology in AML cells with TP53-WT and AML cells with TP53-Y220C. PC14586 in combination with an MDM2 inhibitor, a BCL-2 inhibitor, or an XPO-1 inhibitor is more effective than each of these mono-inhibitors, while PC14586 in combination with any two other activators is more effective than PC14586 in combination with any one other activator. PC14586 in combination with an MDM2 inhibitor, a BCL-2 inhibitor, and an XPO-1 inhibitor is most effective in inducing cell death in both TP53 wild-type AML cells and Y220C mutant AML cells.

[0214] Figure 6A shows the 7AAD / AnnV curves for AML cells with TP53-WT treated for 72 hours with PC14586, a BCL-2 inhibitor, an MDM2 inhibitor, an XPO-1 inhibitor, or a combination of PC14586 and one, two, or all three of these inhibitors. Figure 6B shows the dose-response survival curves for AML cells with TP53-WT treated for 72 hours with PC14586, a BCL-2 inhibitor, an MDM2 inhibitor, an XPO-1 inhibitor, or a combination of PC14586 and one, two, or all three of these inhibitors. Figure 6C shows the 7AAD / AnnV curves for AML cells with TP53-Y220C treated for 72 hours with PC14586, a BCL-2 inhibitor, an MDM2 inhibitor, an XPO-1 inhibitor, or a combination of PC14586 and one, two, or all three of these inhibitors. Figure 6D shows dose-response survival curves of AML cells having TP53-Y220C treated for 72 hours with PC14586, a BCL-2 inhibitor, an MDM2 inhibitor, an XPO-1 inhibitor, or a combination of PC14586 and one, two, or all three of these inhibitors. Consequently, the combination therapies and methods disclosed herein are useful for treating leukemia in subjects where such treatment is necessary.

[0215] Example 8: Molm13 TP53-Y220C cells treated with PC14586 alone or in combination with an XPO-1 inhibitor, MDM2 inhibitor, or BCL-2 inhibitor. PC14586-induced p53 target protein was further upregulated when PC14586 was combined with an XPO-1 inhibitor or an MDM2 inhibitor. Figure 7A shows the results of 24-hour treatment with venetoclax (VEN, 5 nM or 10 nM), nutrin-3a (Nut, 2.5 μM or 5 μM), PC14586 (PC, 2 μM or 4 μM), KPT-8602 (KPT, 100 nM or 200 nM), or a combination of PC14586 and venetoclax (VEN / PC, 5 nM VEN and 2 μM PC, 10 nM VEN and 4 μM PC), a combination of PC14586 and nutrin-3a (Nut / PC, 2.5 μM Nut and 2 μM PC, 5 μM Nut and 4 μM PC), or a combination of PC14586 and KPT-8602 (KPT / PC, 100 nM KPT and 2 μM PC, 200 nM KPT and 4 μM PC). Molm13 TP53-Y220C cells This shows a Western blot at the protein level.

[0216] PC14586 exhibited limited activity in promoting the growth of TP53-Y220C AML cells. However, when combined with a BCL-2 inhibitor, it significantly blocked the transition from G1 to S cells and enhanced apoptosis. These effects were even more pronounced when PC14586 was combined with an XPO-1 inhibitor or an MDM2 inhibitor. Figure 7B shows the cell cycle distribution and apoptosis determined by flow cytometry of cells stained with 5-ethinyl-2'-deoxyuridine (EdU) and DNA dyes. Molm13 TP53-Y220C cells were treated for 72 hours with venetoclax (VEN, 10 nM), nutrin-3a (N3, 5 μM), KPT-8602 (KPT, 200 nM), PC14586 (4 μM), or a combination of PC14586 and venetoclax (4 μM PC14586 and 10 nM VEN), a combination of PC14586 and nutrin-3a (4 μM PC14586 and 5 μM N3), or a combination of PC14586 and KPT (4 μM PC14586 and 200 nM KPT). Figure 7C shows the DNA content and PARP cleavage determined by flow cytometry of Molm13 TP53-Y220C cells treated for 72 hours with venetoclax (VEN, 10 nM), nutrin 3a (N3, 5 μM), KPT-8602 (KPT, 200 nM), PC14586 (4 μM), or a combination of PC14586 and venetoclax (4 μM PC14586 and 10 nM VEN), a combination of PC14586 and nutrin-3a (4 μM PC14586 and 5 μM), or a combination of PC14586 and KPT-8602 (4 μM PC14586 and 200 nM KPT). Cells were stained with EdU, DNA dyes, and antibodies against cleaved PARP.

[0217] Example 9: PC14586 induced cell death in bulk AML cells and stem / progenitor cells in two-thirds of primary AML cell samples containing the Y220C mutation. This combination was highly synergistic in AML cells and stem / progenitor cells, regardless of the response to PC14586 alone. Cells derived from AML patient samples (Figures 8A-8G and 9A-9G), patient-derived xenografts (Figures 10A-10D), or normal myeloid cells (Figures 11A-11D) under mesenchymal stromal cell (MSC) co-culture conditions (COX) were treated for 48 or 96 hours with either PC14586; venetoclax; nutrin-3a; KPT-8062; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. After staining the cells with annexin V / 7-aminoactinomycin D (Ann V / 7-ADD) in the presence of counting beads, cell death and viability were determined by flow cytometry.

[0218] Cells from two patients in Figures 8A–8G and 9A–9G responded to PC14586 as a monotherapy. PC14586 as a monotherapy exhibited limited activity in patient-derived PDX cells in Figures 10A–10D. Combinations of PC14586 with MDM2 inhibitors, BCL-2 inhibitors, and / or XPO-1 inhibitors were synergistic in cells and stem / progenitor cells from all three AML patient samples in Figures 8A–8G, 9A–9G, and 10A–10D. All treatments using primary samples were performed under mesenchymal stromal cell (MSC) co-culture conditions (COX).

[0219] Figures 8A–8G show 47% peripheral blood (PB) samples with 77% of cells having the TP53-Y220C mutation. The samples had a complex cytogenetic composition with mutations in the NF1, JAK2, SMC1A, SRSF2, TET2, and CUX1 genes. Cells were stained with annexin V / 7-aminoactinomycin D (Ann V / 7-ADD) in the presence of counting beads, and then cell death and viability were determined by flow cytometry. Figure 8A shows the CD45+, CD34+, and CD34+CD38- populations 48 hours after treatment with different concentrations of PC14586. Tables 1 and 2 show the EC values ​​for the CD45+, CD34+, and CD34+CD38- cell populations treated with PC14586, respectively.50 Data and IC 50 Provide data.

[0220] (Table 1) EC of different cell types derived from PB samples shown in Figures 8A-8G after being treated with PC14586 for 48 hours 50 TIFF2026522611000078.tif31128

[0221] (Table 2) ICs for different cell types derived from PB samples shown in Figures 8A-8G after being treated with PC14586 for 48 hours 50 TIFF2026522611000079.tif30128

[0222] Figure 8B shows the CD45+ population evaluated by Ann V / 7-ADD+ positive cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. Table 3 provides combination index (CI) values ​​derived from Figure 8B. Figure 8C shows CD45+ viable cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586.

[0223] (Table 3) CI values ​​for different treatment combinations for CD45+ cells TIFF2026522611000080.tif23128

[0224] Figure 8D shows CD34+ as assessed by Ann V / 7ADD+ cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. Table 4 provides combination index (CI) values ​​derived from Figure 8D. Figure 8E shows CD34+ viable cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586.

[0225] (Table 4) CI values ​​for different treatment combinations for CD34+ cells TIFF2026522611000081.tif23128

[0226] Figure 8F shows the CD34+CD38- population evaluated by Ann V / 7ADD+ cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. Table 5 provides combination index (CI) values ​​derived from Figure 8F. Figure 8G shows CD34+CD38- viable cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586.

[0227] (Table 5) CI values ​​for different treatment combinations for CD34+CD38- cells TIFF2026522611000082.tif23128

[0228] Figures 9A–9G show cell samples from AML patients, containing 92% peripheral blood (PB) with 45.7% having TP53-Y220C, under MSC COX. The samples had a complex cytogenetic composition with mutations in TP-53-G105fs (39.7%) and the U2AF1 gene. After staining the cells with Ann V / 7-ADD in the presence of counting beads, cell death and viability were determined by flow cytometry. Figure 9A shows the CD45+, CD34+, and CD34+CD38- populations 48 hours after treatment with different concentrations of PC14586. PC14586 induced cell death in AML blasts and stem / progenitor cells. Tables 6 and 7 show the EC values ​​for the CD45+, CD34+, and CD34+CD38- cell populations treated with PC14586, respectively. 50 Data and IC 50 Provide data.

[0229] (Table 6) EC of different cell types derived from PB samples shown in Figures 9A-9G after being treated with PC14586 for 48 hours 50 TIFF2026522611000083.tif31128

[0230] (Table 7) ICs for different cell types derived from PB samples shown in Figures 9A-9G after being treated with PC14586 for 48 hours 50 TIFF2026522611000084.tif31128

[0231] The combination of PC14586 with BCL-2 inhibition, MDM2 inhibition, and / or XPO-1 inhibition synergistically induced cell death in TP53Y220C mutant AML blast cells. Figure 9B shows the CD45+ population evaluated by Ann V / 7-ADD+ cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. Table 8 provides combination index (CI) values ​​derived from Figure 9B. Figure 9C shows CD45+ viable cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586.

[0232] (Table 8) CI values ​​for different treatment combinations for CD45+ cells TIFF2026522611000085.tif24128

[0233] Figure 9D shows CD34+ as assessed by Ann V / 7ADD+ cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. Table 9 provides combination index (CI) values ​​derived from Figure 9D. Figure 9E shows CD34+ viable cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586.

[0234] (Table 9) CI values ​​for different treatment combinations for CD34+ cells TIFF2026522611000086.tif23128

[0235] Figure 9F shows the CD34+CD38- population evaluated by Ann V / 7ADD+ cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586. Table 10 provides combination index (CI) values ​​derived from Figure 9F. Figure 9G shows CD34+CD38- viable cells 48 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or nutrin-3a, KPT-8602, and PC14586.

[0236] (Table 10) CI values ​​for different treatment combinations for CD34+CD38- cells TIFF2026522611000087.tif24128

[0237] Figures 10A–10D show patient-derived xenograft (PDX) splenocytes, 99.5% of which were hCD45+ and had the TP53-Y220C mutation confirmed by sequencing. The samples had a complex karyotype, MECOM rearrangement, and a complex cytogenetic composition with mutations in the NRAS, KRAS, TP53 Y220C, and P151A genes. After staining the cells with Ann V / 7-ADD in the presence of counting beads, cell death and viability were determined by flow cytometry. Table 11 shows variants that are likely to be of somatic origin.

[0238] (Table 11) Variants in the PDX samples used in Figures 10A-10D that are likely to be of somatic origin TIFF2026522611000088.tif31157

[0239] Figure 10A shows CD45+, CD34+, and CD34+CD38- populations 96 hours after treatment with different concentrations of PC14586. PC14586 as a monotherapy induced limited cell death in AML blasts and stem / progenitor cells.

[0240] The combination of PC14586 with a BCL-2 inhibitor, an MDM2 inhibitor, and / or an XPO-1 inhibitor synergistically induced cell death in TP53Y220C-mutant AML blast cells. Figure 10B shows the CD45+ population evaluated by Ann V / 7-ADD+ positive cells 96 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; nutrin-3a, KPT-8602, and PC14586; venetoclax, KPT-8602, and PC14586; KPT-8602, nutrin-3a, and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586. Table 12 provides combination index (CI) values ​​derived from Figure 10B. Figure 10C: Venetoclax; Nutrin-3a (N3); KPT-8602; PC14586; Venetoclax and PC14586; Nutrin-3a and PC14586; KPT-8602 and PC14586; Nutrin-3a, KPT-8602, and PC14586; Venetoclax, KPT-8602, and PC14586; KPT-8602, Nutrin-3a, and PC14586; or CD45+ viable cells 96 hours after treatment with Venetoclax, Nutrin-3a, KPT-8602, and PC14586.

[0241] (Table 12) CI values ​​for different treatment combinations for CD45+ cells TIFF2026522611000089.tif23128

[0242] Figure 10D shows CD34+ as assessed by Ann V / 7ADD+ cells 96 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; nutrin-3a, KPT-8602, and PC14586; venetoclax, KPT-8602, and PC14586; KPT-8602, nutrin-3a, and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586. Table 13 provides combination index (CI) values ​​derived from Figure 10D. Figure 10E shows CD34+ viable cells 96 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; nutrin-3a, KPT-8602, and PC14586; venetoclax, KPT-8602, and PC14586; KPT-8602, nutrin-3a, and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586.

[0243] (Table 13) CI values ​​for different treatment combinations for CD34+ cells TIFF2026522611000090.tif23128

[0244] Figure 10F shows the CD34+CD38- population evaluated by Ann V / 7ADD+ cells 96 hours after treatment with venetoclax; nutrin-3a (N3); KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; nutrin-3a, KPT-8602, and PC14586; venetoclax, KPT-8602, and PC14586; KPT-8602, nutrin-3a, and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586. Table 14 provides combination index (CI) values ​​derived from Figure 10F. Figure 10G: Venetoclax; Nutrin-3a (N3); KPT-8602; PC14586; Venetoclax and PC14586; Nutrin-3a and PC14586; KPT-8602 and PC14586; Nutrin-3a, KPT-8602, and PC14586; Venetoclax, KPT-8602, and PC14586; KPT-8602, Nutrin-3a, and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586. CD34+CD38- viable cells 96 hours after treatment with these agents.

[0245] (Table 14) CI values ​​for different treatment combinations for CD34+CD38- cells TIFF2026522611000091.tif23128

[0246] Treatment with PC14586, and combinations of treatment with PC14586 and venetoclax, nutrin-3a, and / or KPT-8602, exhibited limited toxicity in normal myeloid cells and CD34+ stem / progenitor cells. Figures 11A–11D show normal myeloid cells under MSC COX. After staining the cells with Ann V / 7-ADD in the presence of counting beads, cell death and viability were determined by flow cytometry. Figure 11A shows the CD45+ population evaluated by Ann V / 7-ADD+ positive cells 48 hours after treatment with venetoclax; nutrin-3a; KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586. Figure 11B shows the CD45+ cell viability 48 hours after treatment with venetoclax; nutrin-3a; KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586. Figure 11C shows the CD34+ population evaluated by Ann V / 7-ADD+ positive cells 48 hours after treatment with venetoclax; nutrin-3a; KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586. Figure 11D shows the CD34+ cell viability 48 hours after treatment with venetoclax; nutrin-3a; KPT-8602; PC14586; venetoclax and PC14586; nutrin-3a and PC14586; KPT-8602 and PC14586; or venetoclax, nutrin-3a, KPT-8602, and PC14586.

[0247] Example 10: PC14586 reactivates p53 signaling, which is further enhanced by inhibition of MDM2 induced by PC14586-activated p53. Molm13 TP53-WT cells and TP53-Y220C cells were treated for 4 hours with Nutrin-3a (N3a, 5 μM), PC14586 (PC, 4 μM), or a combination thereof. RNA was isolated and subjected to RNA sequencing.

[0248] Figure 12A shows principal component analysis comparing Molm13 TP53-WT cells and TP53-Y220C cells treated for 4 hours with Nutrin-3a (N3a, 5 μM), PC14586 (PC, 4 μM), or a combination thereof.

[0249] Figure 12B shows gene set enrichment analysis by 77-step two-dimensional hierarchical clustering comparing Molm13 TP53-WT cells and TP53-Y220C cells treated for 4 hours with nutrin-3a (N3a, 5 μM), PC14586 (PC, 4 μM), or a combination thereof. Arrows in Figure 12B indicate that upregulation of the p53 pathway occurred in Molm13 TP53-WT cells treated with N3a or a combination of N3a and PC14586, but not with PC14586 alone. Arrows in Figure 12B also indicate that upregulation of the p53 pathway occurred in Molm13 TP53-Y220C cells treated with PC14586 or a combination of N3a and PC14586, but not with N3a alone.

[0250] Figure 12C shows a heatmap of genes that were expressed differently. From left to right, Molm13 TP53-Y220C cells were treated with a combination of nutrin-3a and PC14586 (n=3, 5 μM N3a, 4 μM PC); Molm13 TP53-WT cells were treated with nutrin-3a (n=3, N3a, 5 μM); Molm13 TP53-WT cells were treated with a combination of nutrin-3a and PC14586 (n=3, 5 μM N3a, 4 μM PC); Molm13 TP53-Y220C cells were treated with nutrin-3a (n=3, N3a, 5 μM); Molm13 TP53-Y220C cells were treated with vehicle alone as a control (n=3, con); and Molm13 TP53-Y220C cells were treated with PC14586 (n=3, 4 μM PC). TP53-Y220C cells were treated; Molm13 TP53-WT cells were treated with the vehicle alone as a control (n=3, con); and Molm13 TP53-WT cells were treated with PC14586 (n=3, 4 μM PC). The best upregulation occurred in Molm13 TP53-Y220C cells treated with the combination of nutrin-3a and PC14586, and the results showed that no change or disregard occurred in any group to the left of the group of Molm13 TP53-Y220C cells treated with nutrin-3a.

[0251] Figure 12D shows the pathway analysis. Figure 12E shows a volcano plot comparing the increase, remainder, and decrease in gene expression in the treated samples compared to the control sample. Figure 12F shows a Western blot illustrating the changes in gene expression at the protein level.

[0252] Example 11: The combination of PC14586 and RG7388 significantly extended survival in the TP53-Y220C PDX model, while PC14586 alone had no effect in models carrying the TP53-Y220C mutation, the TP53-P151A mutation, and the NAS mutation. PDX cells derived from AML patient samples carrying the TP53-Y220C (VAF 48%) mutation, the TP53-P151A (VAF 47%) mutation, and the NRAS (VAF 50%) mutation were transmitted via the tail vein to NSG mice (8 weeks old, male) (1.6 × 10⁶). 6The drug was injected into each mouse. Mice (10 mice / group) were treated with either the vehicle, PC14586 (100 mg / kg, daily) and the 50% active MDM2 inhibitor idasanutrin (RG7388) (40 mg / kg, initially on for 8 days, then off for 2 days / 5 days) or PC14586 (100 mg / kg, daily) and the 50% active MDM2 inhibitor idasanutrin (RG7388) (40 mg / kg, initially on for 8 days, then off for 2 days / 5 days) when circulating human CD45 positivity reached ≥1% as determined by flow cytometry. Disease progression and treatment response were monitored by flow cytometry of human CD45+ cells. Mouse survival time was estimated using the Kaplan-Meier method, and survival time data were analyzed using the log-rank test.

[0253] Figure 13A shows flow cytometry analysis of human CD45+ cells in peripheral blood (PB) 4 weeks after treatment. Figure 13B shows flow cytometry analysis of human CD45+ cells in peripheral blood (PB) 8 weeks after treatment. Figure 13C: Survival data analyzed using Kaplan-Meier estimation and log-rank test for mouse survival time.

[0254] Example 12: Unlike WTp53, which binds to and antagonizes anti-apoptotic BCL-2, PC14586-activated p53 does not bind to BCL-2. Molm13 TP53-WT cells and TP53-Y220C cells were treated with Nutrin-3a (5 μM) or PC14586 (PC, 1.25 μM or 5 μM) for 4 hours. p53 was co-immunoprecipitation using an antibody selective for the WT conformation of p53 (PAb1620, catalog number 102201, Caprico Bioscience) or an antibody selective for the mutant conformation of p53 (PAb240, NB200-103, Novus Biosciences). After co-immunoprecipitation, p53 and BCL-2 were determined by Western blotting. Figure 14 shows these Western blots of p53 and BCL-2 derived from Molm13 TP53-WT cells and TP53-Y220C cells treated with nutrin-3a (5 μM) or PC14586 (PC, 1.25 μM or 5 μM) for 4 hours. Although not constrained by any theory, the pattern of PC14586-activated p53 that does not bind to BCL-2 may be related to its lack of apoptosis-inducing activity in Molm13 TP53-Y220C cells.

[0255] Example 13: The combination of PC14586 and venetoclax significantly extended survival in the Mol13 TP53-Y220C xenograft model. Molm13 TP53-Y220C cells 0.5×10 6 Individual cells / mouse were injected into NSG mice (male, 6-10 weeks old) via the tail vein. Mice (5 / group) were treated daily with vehicle, PC14586 (100 mg / kg), venetoclax (50 mg / kg), or a combination. Mouse survival was estimated using the Kaplan-Meier method, and survival data were analyzed using the log-rank test. Figure 15 shows survival data for mice injected with Molm13 TP53-Y220C cells and treated daily with vehicle, PC14586, venetoclax, or a combination, analyzed using Kaplan-Meier estimation and the log-rank test.

[0256] Example 14: The combination of PC14586 and an XPO-1 inhibitor greatly increased the nuclear localization and transcriptional activity of p53. TP53-Y220C Molm13 cells were treated with PC14586 (4 μM), KPT-8602 (200 nM), or both for 24 hours. p53 localization and its target proteins were determined by Western blotting (MWM, molecular weight markers). For protein localization, cytoplasmic and nuclear proteins were fractionated. Tubulin and lamin B1 were used as cytoplasmic and nuclear loading controls, respectively. Figure 16 shows Western blots of p53, lamin B1, and tubulin from TP53-Y220C Molm13 cells treated with PC14586 (4 μM), KPT-8602 (200 nM), or both for 24 hours.

[0257] Equal portions This technology is not limited to the specific embodiments described herein, which are merely illustrative of the individual aspects of the technology. As will be apparent to those skilled in the art, many modifications and variations of this technology can be made without departing from its spirit and scope. In addition to those enumerated herein, functionally equivalent methods and apparatus within the scope of this technology will be apparent to those skilled in the art from the foregoing description. Such modifications and variations are intended to be included within the scope of this technology. This technology is not limited to specific methods, reagents, compound compositions, or biological systems, which can, of course, be modified. It will also be understood that the terminology used herein is for the purpose of describing specific embodiments only and is not intended to limit them.

[0258] Furthermore, if any feature or aspect of this disclosure is described in relation to the Markush group, a person skilled in the art will recognize that this disclosure is also described in relation to any individual member or subgroup of members of the Markush group.

[0259] For all purposes, specifically with regard to providing written explanations, it will be understood by those skilled in the art that all scopes disclosed herein also encompass all possible sub-scopes and combinations of sub-scopes. Any listed scope can be readily recognized as sufficient to explain and enable that the same scope can be divided into at least two, three, four, five, ten, and so on. As a non-limiting example, each scope discussed herein can easily be divided into a lower third, a middle third, an upper third, and so on. It will also be understood by those skilled in the art that all words such as “at most,” “at least,” “greater than,” “less than,” and similar terms include the number described and refer to a scope that can subsequently be divided into the sub-scopes discussed above. Finally, it will be understood by those skilled in the art that a scope includes each individual member. Thus, for example, a group having 1 to 3 cells refers to a group having 1, 2, or 3 cells. Similarly, groups having 1 to 5 cells refer to groups having 1, 2, 3, 4, or 5 cells, and so on.

[0260] All patents, patent applications, provisional applications, and publications referenced or cited herein, including all figures and tables, are incorporated by reference to the extent that they do not contradict the express teachings herein.

Claims

1. A method for treating wild-type p53 leukemia or TP53-Y220C leukemia in patients who require it, The process includes administering to the patient an effective amount of a TP53 reactivated indole derivative and an effective amount of an MDM2 inhibitor, a BCL-2 inhibitor, and / or an XPO-1 inhibitor. The TP53 reactivated indole derivative is of formula (I): Having the formula, or a pharmaceutically acceptable salt thereof, During the ceremony, each However, they are independently single or double bonds; X 1 However, CR 5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 2 is a carbon atom linked to 7 CR 7 R 8 NR 7 O, S, C=O, C=S, or Q 1 ; and X 3 However, CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 4 However, CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 5 However, CR 13 , N, or NR 13 And; X 1 , X 2 , X 3 , and X 4 At least one of them is Q 1 It is a carbon atom linked to; Q 1 However, C=O, C=S, C=CR 14 R 15 , C=NR 14 , alkylene, alkenylene, or alkynylene (each of these independently substituted or unsubstituted), bond; m is 1, 2, 3, or 4; Y is either N or O; R 1 However, -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 ,-R 16 Ure 17 ,-R 16 Ure 17 R 18 , -SR 16 ,-R 16 SR 17 ,-R 16 SR 17 R 18 , -NR 16 R 17 ,-R 16 NR 17 ,-R 16 NR 17 R 18 , -NR 16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 However, independently, -C(O)R 19 , -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 , -SO 2 R 19 , alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently unsubstituted or substituted), or hydrogen; Each R 2 、R 5 、R 6 、R 7 、R 8 、R 9 、R 10 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、and R 18 is, independently, -C(O)R 21 、-C(O)OR 21 、-C(O)NR 21 R 22 、-OR 21 、-SR 21 、-NR 21 R 22 、-NR 21 C(O)R 22 、-OC(O)R 21 、halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently unsubstituted or substituted), or hydrogen; Each R 19 and R 20 However, C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R 21 and R 22 However, independently, they are alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R 23 and R 24 However, independently, they are alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently substituted or unsubstituted), or hydrogen. The method.

2. A method for extending the survival period of patients with wild-type p53 leukemia or TP53-Y220C leukemia, The procedure includes administering to a leukemia patient an effective amount of a TP53 reactivated indole derivative, and an effective amount of at least one additional therapeutic substance selected from MDM2 inhibitors, BCL-2 inhibitors, and XPO-1 inhibitors. The TP53 reactivated indole derivative is of formula (I): Having the formula, or a pharmaceutically acceptable salt thereof, During the ceremony, each However, they are independently single or double bonds; X 1 However, CR 5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 2 However, CR 7 CR 7 R 8 , N, NR 7 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 3 However, CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 4 However, CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 5 However, CR 13 , N, or NR 13 And; X 1 , X 2 , X 3 , and X 4 At least one of them is Q 1 It is a carbon atom linked to; Q 1 However, C=O, C=S, C=CR 14 R 15 , C=NR 14 , alkylene, alkenylene, or alkynylene (each of these independently substituted or unsubstituted), bond; m is 1, 2, 3, or 4; Y is either N or O; R 1 However, -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 ,-R 16 Ure 17 ,-R 16 Ure 17 R 18 , -SR 16 ,-R 16 SR 17 ,-R 16 SR 17 R 18 , -NR 16 R 17 ,-R 16 NR 17 ,-R 16 NR 17 R 18 , -NR 16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 However, independently, -C(O)R 19 , -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 , -SO 2 R 19 , alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently unsubstituted or substituted), or hydrogen; Each R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 However, independently, -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 , halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently unsubstituted or substituted), or hydrogen; Each R 19 and R 20 However, C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R 21 and R 22 However, independently, they are alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R 23 and R 24 However, independently, they are alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently substituted or unsubstituted), or hydrogen. The method.

3. A method for selecting leukemia patients for treatment with TP53 reactivated indole derivatives and additional therapeutic substances, A step of detecting the expression of wild-type TP53 or TP53-Y220C mRNA or polypeptide in biological samples obtained from leukemia patients; and The procedure includes administering to a leukemia patient an effective amount of a TP53 reactivated indole derivative, and an effective amount of at least one additional therapeutic substance selected from MDM2 inhibitors, BCL-2 inhibitors, and XPO-1 inhibitors. The TP53 reactivated indole derivative is of formula (I): Having the formula, or a pharmaceutically acceptable salt thereof, During the ceremony, each However, they are independently single or double bonds; X 1 However, CR 5 CR 5 R 6 , N, NR 5 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 2 However, CR 7 CR 7 R 8 , N, NR 7 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 3 However, CR 9 CR 9 R 10 , N, NR 9 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 4 However, CR 11 CR 11 R 12 , N, NR 11 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 5 However, CR 13 , N, or NR 13 And; X 1 , X 2 , X 3 , and X 4 At least one of them is Q 1 It is a carbon atom linked to; Q 1 However, C=O, C=S, C=CR 14 R 15 , C=NR 14 , alkylene, alkenylene, or alkynylene (each of these independently substituted or unsubstituted), bond; m is 1, 2, 3, or 4; Y is either N or O; R 1 However, -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 ,-R 16 Ure 17 ,-R 16 Ure 17 R 18 , -SR 16 ,-R 16 SR 17 ,-R 16 SR 17 R 18 , -NR 16 R 17 ,-R 16 NR 17 ,-R 16 NR 17 R 18 , -NR 16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 However, independently, -C(O)R 19 , -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 , -SO 2 R 19 , alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently unsubstituted or substituted), or hydrogen; Each R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 However, independently, -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 , halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently unsubstituted or substituted), or hydrogen; Each R 19 and R 20 However, C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R 21 and R 22 However, independently, they are alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R 23 and R 24 However, independently, they are alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently substituted or unsubstituted), or hydrogen. The method.

4. The method according to any one of claims 1 to 3, wherein the step of administering to the leukemia patient includes the step of administering an effective amount of the TP53 reactivated indole derivative and an effective amount of the MDM2 inhibitor.

5. The method according to any one of claims 1 to 3, wherein the step of administering to the leukemia patient includes the step of administering an effective amount of the TP53 reactivated indole derivative and an effective amount of the BCL-2 inhibitor.

6. The method according to any one of claims 1 to 3, wherein the step of administering to the leukemia patient includes the step of administering an effective amount of the TP53 reactivated indole derivative and an effective amount of the XPO-1 inhibitor.

7. The method according to any one of claims 1 to 3, wherein the step of administering to the leukemia patient includes administering an effective amount of the TP53 reactivated indole derivative, an effective amount of the MDM2 inhibitor, and an effective amount of the BCL-2 inhibitor.

8. The method according to any one of claims 1 to 3, wherein the step of administering to the leukemia patient includes administering an effective amount of the TP53 reactivated indole derivative, an effective amount of the MDM2 inhibitor, and an effective amount of the XPO-1 inhibitor.

9. The method according to any one of claims 1 to 3, wherein the step of administering to the leukemia patient includes administering an effective amount of the TP53 reactivated indole derivative, an effective amount of the BCL-2 inhibitor, and an effective amount of the XPO-1 inhibitor.

10. The method according to any one of claims 1 to 3, wherein the step of administering to the leukemia patient includes administering an effective amount of the TP53 reactivated indole derivative, an effective amount of the MDM2 inhibitor, an effective amount of the BCL-2 inhibitor, and an effective amount of the XPO-1 inhibitor.

11. The method according to any one of claims 1 to 10, wherein the MDM2 inhibitor is selected from the group consisting of RG7112, RO5045337, idasanutrin, nutrin-3a, RG7388, AMG-232, KRT-232, APG-115, BI-907828, CGM097, cilemadrin, HDM201, mirademethane, MEL23, MEL24, and DS-3032b.

12. The method according to any one of claims 1 to 11, wherein the BCL-2 inhibitor is selected from the group consisting of venetoclax, ovatoclax, subatoclax, maritoclax, gossypol, apogossypol, TW-37, UMI-77, and BDA-366.

13. The method according to any one of claims 1 to 12, wherein the XPO-1 inhibitor is selected from the group consisting of KPT330, XPOVIO, KPT8602 (ertanexol), KPT335, verginexol, and KPT185.

14. The method according to any one of claims 1 to 13, wherein the MDM2 inhibitor, the BCL-2 inhibitor, and / or the XPO-1 inhibitor are administered sequentially, simultaneously, or separately with the TP53 reactivating indole derivative.

15. The method according to any one of claims 1 to 14, wherein the MDM2 inhibitor, the BCL-2 inhibitor, the XPO-1 inhibitor, or the TP53 reactivated indole derivative is administered orally, intravenously, intramuscularly, intraperitoneally, or subcutaneously.

16. The method according to any one of claims 1 to 15, wherein the leukemia is acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS).

17. The method according to any one of claims 1 to 16, wherein the leukemia is TP53-Y220C leukemia.

18. The TP53 reactivated indole derivative is of formula (IA): Having the formula, or a pharmaceutically acceptable salt thereof, During the ceremony, X 1 However, CR 5 , NR 5 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 2 However, CR 7 CH, NR 7 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 3 However, CR 9 CH, NR 9 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 4 However, CR 11 CH, NR 11 , O, S, C=O, C=S, or Q 1 It is a carbon atom linked to; X 5 However, CR 13 CH, or NR 13 And; X 1 , X 2 , X 3 , and X 4 At least one of them is Q 1 It is a carbon atom linked to; Q 1 However, it is a combination; m is 1, 2, 3, or 4; Y is either N or O; R 1 However, -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 ,-R 16 Ure 17 ,-R 16 Ure 17 R 18 , -SR 16 ,-R 16 SR 17 ,-R 16 SR 17 R 18 , -NR 16 R 17 ,-R 16 NR 17 ,-R 16 NR 17 R 18 , -NR 16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 However, independently, -C(O)R 19 , -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 , -SO 2 R 19 , hydrogen, alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl, where each of alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl is independently unsubstituted, or substituted with one or two groups selected from alkyl or halogen, or R 3 and R 4 However, R 3 and R 4 It forms a ring together with the bonded Y atom, and here, the ring is substituted or unsubstituted or R 3 However, he was absent; Each R 2 , R 5 , R 7 , R 9 , R 11 , R 13 , R 16 , R 17 , and R 18 However, independently, -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 , hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, where each alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl is independently unsubstituted or -C(O)R 25 , -C(O)OR 25 -C(O)NR 25 ,-CR 25 3 , -OR 25 , -SR 25 , -NR 25 R 26 , -NR 25 C(O)R 26 -OC(O)R 25 , substituted with one or two groups selected from alkyl, alkenyl, or alkynyl, R 25 and R 26 Each of these is independently an alkyl, hydrogen, or halogen; Each R 19 and R 20 However, C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R 21 and R 22 However, independently, they are alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R 23 and R 24 However, independently, they are alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently substituted or unsubstituted), or hydrogen. The method according to any one of claims 1 to 17.

19. The aforementioned TP53 reactivated indole derivative is of formula (IB): Having the formula, or a pharmaceutically acceptable salt thereof, During the ceremony, X 1 However, CR 5 , NR 5 , a carbon atom linked with O, S, C=O, C=S, or Y; X 2 However, CR 7 CH, NR 7 , a carbon atom linked with O, S, C=O, C=S, or Y; X 3 However, CR 9 CH, NR 9 , a carbon atom linked with O, S, C=O, C=S, or Y; X 4 However, CR 11 CH, NR 11 , a carbon atom linked with O, S, C=O, C=S, or Y; X 5 However, CR 13 CH, or NR 13 And; X 1 , X 2 , X 3 , and X 4 At least one of them is a carbon atom linked to Y; Y is either N or O; R 1 However, -C(O)R 16 , -C(O)OR 16 -C(O)NR 16 R 17 , -OR 16 ,-R 16 Ure 17 ,-R 16 Ure 17 R 18 , -SR 16 ,-R 16 SR 17 ,-R 16 SR 17 R 18 , -NR 16 R 17 ,-R 16 NR 17 ,-R 16 NR 17 R 18 , -NR 16 C(O)R 16 -OC(O)R 16 , C=O, C=S, -CN, -SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; Each R 3 and R 4 However, independently, -C(O)R 19 , -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 , -SO 2 R 19 , hydrogen, alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl, where each of alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl is independently unsubstituted, or substituted with one or two groups selected from alkyl or halogen, or R 3 and R 4 However, R 3 and R 4 It forms a ring together with the bonded Y atom, and here, the ring is substituted or unsubstituted or R 3 However, he was absent; Each R 2 , R 5 , R 7 , R 9 , R 11 , R 13 , R 16 , R 17 , and R 18 However, independently, -C(O)R 21 , -C(O)OR 21 -C(O)NR 21 R 22 ,-CR 25 3 , -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 -OC(O)R 21 , hydrogen, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, where each alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl is independently unsubstituted or -C(O)R 25 , -C(O)OR 25 -C(O)NR 25 ,-CR 25 3 , -OR 25 , -SR 25 , -NR 25 R 26 , -NR 25 C(O)R 26 -OC(O)R 25 , substituted with one or two groups selected from alkyl, alkenyl, or alkynyl, R 25 and R 26 Each of these is independently an alkyl, hydrogen, or halogen; Each R 19 and R 20 However, C(O)R 23 , -C(O)OR 23 -C(O)NR 23 R 24 , -OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen or halogen; Each R 21 and R 22 However, independently, they are alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of which is independently substituted or unsubstituted), or hydrogen; and Each R 23 and R 24 However, independently, they are alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl (each of these independently substituted or unsubstituted), or hydrogen. The method according to any one of claims 1 to 17.

20. The aforementioned TP53 reactivated indole derivative is given by formula: A method according to any one of claims 1 to 17, comprising:

21. The method according to any one of claims 3 to 20, wherein mRNA expression levels are detected by real-time quantitative PCR (qPCR), digital PCR (dPCR), reverse transcriptase-PCR (RT-PCR), Northern blotting, microarrays, dot or slot blotting, in-situ hybridization, or fluorescence in-situ hybridization (FISH).

22. The method according to any one of claims 3 to 21, wherein polypeptide expression levels are detected by Western blotting, enzyme-linked immunosorbent assay (ELISA), dot blotting, immunohistochemistry, immunofluorescence, immunoprecipitation, immunoelectrophoresis, or mass spectrometry.

23. The method according to any one of claims 1 to 22, wherein the patient is unresponsive to at least one prior cancer treatment.

24. The method according to claim 23, wherein the at least one prior cancer treatment is chemotherapy or immunotherapy.

25. The aforementioned chemotherapy regimens include trioxide, azacitidine, cerubidine, cyclophosphamide, cytarabine, daunorubicin hydrochloride, daurismo, dexamethasone, doxorubicin hydrochloride, enasidenib mesylate, gemtuzumab ozogamicin, gilteritinib fumarate, glassegib maleate, idamycin PFS, idarubicin hydrochloride, idhifa, ivosidenib, midostaurine, mitoxantrone hydrochloride, mylotarg, and ortasidenib. The method according to claim 24, comprising one or more of the following: nib, onureg, pemazyre, pemigatinib, prednisone, rezlidhia, rituxan, rituximab, rubidomycin, rydapt, tabloid, thioguanine, tibsovo, tisagenlecleucel, trisenox, venclexta, venetoclas, vinicristine sulfate, vyxeos, or xospata.

26. The method according to any one of claims 1 to 25, wherein the patient is a child or an adult.

27. The method according to any one of claims 3 to 26, wherein the biological sample is whole blood, serum, or plasma.

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