KR20210103973A - Protein Kinase Degradation Inducing Compounds, and Use thereof - Google Patents

Abstract

The present invention relates to a protein kinase degradation inducing compound and uses thereof. Specifically, the protein kinase degradation inducing compound of the present invention can degrade AURKA, FLT3 mutant, EGFR mutant, or c-KIT mutant protein, and thereby being able to treat diseases related to AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation.

Description

Protein Kinase Degradation Inducing Compounds and Their Uses {Protein Kinase Degradation Inducing Compounds, and Use thereof}

The present invention relates to a protein kinase degradation inducing compound, a method for preparing the same, and a use thereof. Specifically, the present invention relates to AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutagenesis-inducing compound and a method for preparing the same, and further relates to AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutagenesis through activity It relates to use for the treatment of disease.

Anticancer drugs have been developed into cytotoxic anticancer drugs (1st generation), targeted anticancer drugs (2nd generation), and immune anticancer drugs (3rd generation). In particular, the target anticancer agent is a protein kinase inhibitor, and the type of kinase inhibited and the presence or absence of a mutation determine the type of cancer that can be treated. For example, AURKA (Aurora A kinase) is a target of acute lymphoblastic leukemia (ALL) or other carcinomas, and FLT3 (Fms-like tyrosine kinase 3) is a gene frequently mutated in patients with acute myeloid leukemia (AML). For one, the FLT3 (ITD) mutation is a target for acute myeloid leukemia (AML) or other carcinomas. The c-KIT (D816V) mutation, a point mutation in exon 17 of the c-KIT gene, is a target for ALL or other carcinomas and is also a target for systemic mastocytosis (SM). In addition, Del19, C797S, Del19/C797S, T790M/C797S, and Del19/T790M/C797S mutations reported in the EGFR (Epidermal growth factor receptor) gene are known to be targets of various carcinomas such as lung cancer, colorectal cancer, glioblastoma, and head and neck cancer. have. However, these protein kinase inhibitors do not have sufficient activity to show a significant therapeutic effect in clinical practice or have a problem with side effects due to toxicity, so a technology capable of solving these problems is desired.

As described above, the low molecular weight compounds so far have been focused on inhibiting the function of disease-related proteins, but as a new concept, a selective proteolysis method that removes the disease-related protein itself is emerging. In particular, a ubiquitin-proteasome system (Ubiquitin-Proteasome System) using PROTAC (Proteolysis targeting chimera) is attracting attention.

PROTAC is a bifunctional small molecule in which a ligand molecule that binds to a disease-related target protein and an E3 ubiquitin ligase binding moiety are linked by a chemical linker. PROTAC technology can target undruggable proteins that cannot be targeted by existing drugs, overcome the phenomenon of drug resistance being expressed or a decrease in drug efficacy caused by activation of other signal transduction systems, and the binding and degradation process Target selectivity is improved because the target protein is removed through There are several advantages that can be achieved. Therefore, PROTAC technology is receiving great attention as a new drug development platform technology for low molecular weight compounds.

Recent Pat Anticancer Drug Discov. 2008, 3(3), 162-177 Leukemia, 2019, 33, 299-312 Expert Opin. Investing. Drugs, 2013, 22(1), 103-115 Ito, Takumi, et al. Science 　327.5971 (2010): 1345-1350. Chamberlain, Philip P., and Brian E. Cathers. Drug Discovery Today: Technologies (2019). Akuffo, Afua A., et al. Journal of Biological Chemistry 293.16 (2018): 6187-6200.] Burslem, George M., et al. ChemMedChem 13.15 (2018): 1508-1512. Schneekloth, John S., et al. Journal of the American Chemical Society 126.12 (2004): 3748-3754. Rodriguez-Gonzalez, A., et al. Oncogene 27.57 (2008): 7201. Buckley, Dennis L., et al. Journal of the American Chemical Society 134.10 (2012): 4465-4468. Buckley, Dennis L., et al. Angewandte Chemie International Edition 51.46 (2012): 11463-11467. Galdeano, Carles, et al. Journal of medicinal chemistry 57.20 (2014): 8657-8663. Soares, Pedro, et al. Journal of medicinal chemistry 61.2 (2017): 599-618.

It is an object of the present invention to provide a protein kinase degradation inducing compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. Another object of the present invention is to provide a method for producing a protein kinase degradation inducing compound. Another object of the present invention is to provide the use of a protein kinase degradation inducing compound, specifically, AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation-related disease comprising the compound as an active ingredient. A composition for treatment or prevention , AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation-related disease using the compound for the treatment or prevention of disease or AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation-related disease comprising administering the compound We want to provide a treatment method for

As a result of research efforts by the present inventors to achieve the above object, it was confirmed that the protein kinase degradation inducing compound represented by Formula 1 mentioned below degrades AURKA, FLT3 mutant, EGFR mutant, or c-KIT mutant protein, and the present invention was completed.

Protein kinase degradation inducing compounds

The present invention provides novel compounds that induce degradation of AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation. Specifically, the present invention provides a protein kinase degradation-inducing compound represented by the following formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Formula 1,

이고,TL (Target Ligand) is

ego,

는 Linker와 공유결합으로 연결됨을 나타냄),ELL (E3 Ligase Ligand) is an E3 ubiquitin ligase ligand selected from the group consisting of the following formulas A-1, B-1 and C-1 (wherein

indicates that it is covalently linked with the Linker),

[Formula A-1]

[Formula B-1]

{In the above formulas A-1 and B-1,

는

,

,

,

,

및

로 구성된 군에서 선택된 고리이고;

Is

,

,

,

,

and

a ring selected from the group consisting of;

X ₁ is a single bond, -C≡C-, -CH ₂ -, -NH-, -O-, -CO-, -COO-, -NHCO- or -CONH-;

X ₂ is -CH ₂ -, -CH(C _1-4 alkyl)-, -NH-, -N(C _1-4 alkyl)-, -O-, -CO-, -CH ₂ -CH ₂ -, -NH-CH ₂ -, -NH-CH(C _1-4 alkyl)-, -N=CH-, or -N=C(C _1-4 alkyl)-;

X ₃ is hydrogen or C _1-4 alkyl;

X ₄ is hydrogen, halogen, C _1-6 alkyl, CN, NH ₂ , NO ₂ , OH, COH, COOH or CF ₃ ,

X ₅ is -CO- or -CH ₂ -}

[Formula C-1]

{In the formula C-1,

n is an integer from 1 to 3;

는 C_5-6사이클로알킬, 페닐, 5 내지 6원 헤테로사이클로알킬 또는 헤테로아릴이고{상기 헤테로사이클로알킬 또는 헤테로아릴은 N, O 또는 S 원자를 1 내지 3개 포함함};

is C _5-6 cycloalkyl, phenyl, 5-6 membered heterocycloalkyl or heteroaryl, wherein said heterocycloalkyl or heteroaryl contains 1 to 3 N, O or S atoms;

Y ₁ is hydrogen or C _1-4 alkyl;

Y ₂ is C _1-4 alkyl, hydroxy(C _1-4 alkyl), —(C _0-2 alkyl)-COH, C _3-8 cycloalkyl, or phenyl;

또는

이고{여기서,

는 Y₃가 상기 화학식 C-1에 공유결합으로 연결되는 위치를 나타냄};Y ₃ is hydrogen,

or

and {here,

represents a position at which Y ₃ is covalently linked to Formula C-1;

Y ₄ is hydrogen, halogen, C _1-4 alkyl, —O(C _1-4 alkyl), C _3-6 cycloalkyl or 4-6 membered heterocycloalkyl [wherein one hydrogen of _{Y 4 is halogen,} -OH, -CN, -NHCOH, -NHCOCH ₃ , -COH or -COCH ₃ may be substituted];

Y ₅ is hydrogen or C _1-4 alkyl;

는 5 내지 12원 헤테로사이클로알킬 또는 헤테로아릴이고{상기 헤테로사이클로알킬은 N 원자를 1개 이상 포함함};

is 5 to 12 membered heterocycloalkyl or heteroaryl, wherein said heterocycloalkyl contains at least one N atom;

Y ₆ is C _1-6 alkyl, halogen or =O};

Linker is a device that connects TL and ELL.

Hereinafter, the target protein ligand (TL), the E3 ligase ligand (ELL) and the linker, which are moieties constituting the compound of the present invention, will be looked at in more detail.

(1) target protein ligand (TL)

이다. 상기 화합물은 일반명이 스타우로스포린(Staurosporine)이며, IUPAC 명칭은 (9S,10R,11R,13R)-2,3,10,11,12,13-Hexahydro-10-methoxy-9-methyl-11-(methylamino)-9,13-epoxy-1H,9H-diindolo[1,2,3-gh:3',2',1'-lm]pyrrolo[3,4-j][1,7]benzodiazonin-1-one인 인돌로카바졸 유도체이다. 스타우로스포린은 Streptomyces staurosporeus가 생산하는 미생물 알칼로이드의 일종으로, 항진균 또는 항고혈압 활성이 있음이 알려져 있으나 임상적으로 사용되지는 않고 있다. 스타우로스포린은 공지된 제법으로 합성하거나 미생물로부터 수득할 수 있고, 유통 중인 물질을 사용할 수도 있다.In the compound represented by Formula 1 of the present invention, TL, which is a moiety performing a target protein ligand function, is

am. The compound has a generic name of Staurosporine, and the IUPAC name is (9S,10R,11R,13R)-2,3,10,11,12,13-Hexahydro-10-methoxy-9-methyl-11- (methylamino)-9,13-epoxy-1H,9H-diindolo[1,2,3-gh:3',2',1'-lm]pyrrolo[3,4-j][1,7]benzodiazonin- It is a 1-one indolocarbazole derivative. Staurosporin is a kind of microbial alkaloid produced by Streptomyces staurosporeus . It is known to have antifungal or antihypertensive activity, but has not been used clinically. Staurosporin may be synthesized by a known method or may be obtained from a microorganism, or a commercially available material may be used.

(2) E3 ligase ligand (ELL)

According to one embodiment of the present invention, ELL is a CRBN E3 ubiquitin ligase binding moiety represented by formula A-1. In the present invention, CRBN means Cereblon E3 ubiquitin ligase. CRBN together with DDB1, Cul4A and ROC1 constitute the E3 ubiquitin ligase complex, where CRBN is the substrate recognition subunit of the complex. Some compounds capable of binding to CRBN E3 ubiquitin ligase are known in the art. For example, after it was known that thalidomide binds to CRBN E3 ubiquitin ligase (Ito et al. 2010), a number of imide-based small molecule compounds including lenalidomide and pomalidomide (immunomodulatory imide) drug; IMiD) has been reported to have CRBN binding capacity (Chamberlain and Brian. 2019, Akuffo et al. 2018), Burslem et al. 2018], etc.).

In one embodiment of the present invention, the CRBN E3 ubiquitin ligase ligand represented by Formula A-1 is represented by Formula A-2 below.

[Formula A-2]

In Formula A-2,

X ₂ is —CH ₂ —, —CH(C _1-4 alkyl)- or —CO—;

X ₃ is hydrogen or C _1-4 alkyl.

In one embodiment of the present invention, Formula A-2 may be selected from the group consisting of the following moieties.

An example of the CRBN E3 ubiquitin ligase ligand according to the present invention is as follows (Chamberlain and Brian. 2019 and Akuffo et al. 2018).

Another example of the CRBN E3 ubiquitin ligase binding moiety according to the present invention is as follows (Burslem et al. 2018).

According to another embodiment of the present invention, ELL is a CRBN E3 ubiquitin ligase binding moiety represented by formula B-1. This moiety is a structural variant of cereblon E3 ubiquitin ligase that exhibits an E3 ubiquitin binding ability similar to and equivalent to the cereblon E3 ubiquitin ligase represented by Formula A-1 described above.

In one embodiment of the present invention, the modified CRBN E3 ubiquitin ligase ligand represented by Formula B-1 is represented by Formula B-2 below.

[Formula B-2]

In Formula B-2,

X ₂ is -NH- or -N(C _1-4 alkyl)-;

X ₃ is hydrogen or C _1-4 alkyl.

In one embodiment of the present invention, Formula B-2 may be selected from the group consisting of the following moieties.

An example of the modified CRBN E3 ubiquitin ligase ligand according to the present invention is as follows (see International Publication No. WO2019/060693 A1, etc.).

According to another embodiment of the present invention, the ELL is a VHL E3 ubiquitin ligase ligand represented by the above formula (C-1). In the present invention, VHL means von Hippel-Lindau tumor suppressor. VHL together with Elongin B, Elongin C, CUL2 and Rbx1 constitute the VCB E3 ligase complex, where VHL is the substrate recognition subunit of the complex. Some compounds capable of binding to VHL E3 ubiquitin ligase are known in the art. For example, Ala-Leu-Ala-(Hy)Pro-Tyr-Ile-Pro heptapeptide (Schneekloth et al. 2004), Leu-Ala-(Hy)Pro-Tyr-Ile pentapeptide (Rodriguez- After peptides such as Gonzalez et al. 2008) were known, a small molecule VHL E3 ubiquitin ligase-binding compound that improved it was reported (Buckley et al. J. Am. Chem. Soc. 2012), literature [Buckley et al. Ang. Chem. Int. Ed. 2012], Galdeano et al. 2014, Soares et al. 2017, etc.).

In one embodiment of the present invention, the VHL E3 ubiquitin ligase ligand is a compound represented by Formula C-1. The definition of Formula C-1 is as described above.

이고, 구체적으로In one embodiment, in Formula C-1, Y ₃ is

and specifically

또는

이다.

or

am.

이고, 구체적으로

또는

이다.In one embodiment, in Formula C-1, Y ₃ is

and specifically

or

am.

In one embodiment of the present invention, the VHL E3 ubiquitin ligase ligand is a compound selected from the group consisting of the following formulas C-2-1 and C-2-2.

[Formula C-2-1]

[Formula C-2-2]

In the above formulas C-2-1 and C-2-2,

는 옥사졸, 이소옥사졸, 싸이아졸, 이소싸이아졸, 이미다졸, 피라졸, 트리아졸, 옥사디아졸, 피롤, 피롤리딘, 퓨란, 디하이드로퓨란 및 테트라하이드로퓨란으로 구성된 군에서 선택된 5원 헤테로아릴 고리이고;

is a 5-membered member selected from the group consisting of oxazole, isoxazole, thiazole, isothiazole, imidazole, pyrazole, triazole, oxadiazole, pyrrole, pyrrolidine, furan, dihydrofuran and tetrahydrofuran a heteroaryl ring;

Y ₁ is hydrogen or C _1-3 alkyl;

Y _{4 is hydrogen or C 1-4} alkyl or C _3-5 cycloalkyl which may be substituted with halogen.

In one embodiment, the VHL E3 ubiquitin ligase represented by Formula C-2-1 is selected from the following group.

In one embodiment, the VHL E3 ubiquitin ligase represented by Formula C-2-2 is selected from the following group.

Another example of the VHL E3 ubiquitin ligase ligand according to the present invention is as follows (Galdeano et al. (2014)).

Another example of the VHL E3 ubiquitin ligase ligand according to the present invention is as follows (Soares et al. 2017).

를 통해 Linker가 공유결합으로 연결될 수 있고,

가 표시되지 않은 E3 유비퀴틴 라이게이즈 리간드 화합물의 경우, 1개의 수소가 단일결합으로 치환되어 Linker와 연결될 수 있다.In the present invention, a linker may be attached to the E3 ubiquitin ligase ligand (ELL) at a position necessary to exert the dual function of PROTAC. In the present invention, represented by formulas A-1, A-2, B-1, B-2, C-1, C-2-1, C-2-2

Linkers can be covalently linked through

In the case of an E3 ubiquitin ligase ligand compound in which is not indicated, one hydrogen may be substituted with a single bond to link with a Linker.

(3) Linker

According to one embodiment of the present invention, Linker is represented by the following formula L-1.

[Formula L-1]

In the above formula,

로 구성된 군에서 선택된 기이고{여기서, 상기 기 내 수소는 할로겐, 히드록시, CF₃, C_1-6알킬 또는 C_3-6사이클로알킬로 치환될 수 있음};L _i is each independently a single bond, -CH ₂ -, -NH-, -O-, -S-, -SO-, -SO ₂ -, -CO-, -CH ₂ CH ₂ -, -CHCH-, -C≡C-, -CH ₂ CH ₂ O-, _{-OCH 2} CH ₂ -, -CH ₂ CH ₂ S-, -SCH ₂ CH ₂ -, -COO-, -CONH-, -NHCO- and

is a group selected from the group consisting of {wherein hydrogen in the group may be substituted with halogen, hydroxy, CF ₃ , C _1-6 alkyl or C _3-6 cycloalkyl};

은 3원 내지 14원 사이클로알킬, 헤테로사이클로알킬, 아렌 또는 헤테로아렌이고;

is 3-14 membered cycloalkyl, heterocycloalkyl, arene or heteroarene;

p is an integer from 0 to 30;

In one embodiment, the formula L-1 is represented by the following formula L-2.

[Formula L-2]

In the above formula,

를 통해 화학식 1에 정의된 TL 모이어티와 공유 결합하고, L_E는 이에 연결된

를 통해 화학식 1에 정의된 ELL 모이어티와 공유 결합하며, L_T는 및 L_E는 각각 독립적으로 -L_X1- 또는 -L_X1-L_C-L_X2-이고,L _T is connected to it

Via a TL moiety covalently bonded defined in formula (I) and, L _E is associated with it

covalently bonded to the ELL moiety as defined in formula 1 through L _T and L _E are each independently -L _X1 - or -L _X1 -L _C -L _X2 -

L _X1 and L _X2 are each independently a single bond, -CH ₂ -, -O-, -NH-, -CO-, -NHCO- or -CONH-,

L _C is C _1-10 alkyl, C _6-14 arene, 5-14 membered heteroarene, C _3-12 cycloalkyl or 5-14 membered heterocycloalkyl {wherein said alkyl, arene, heteroarene, cycloalkyl or heterocycloalkyl may be substituted with halogen, C _1-6 alkyl, C _1-6 haloalkyl, C _1-5 alkoxy, C _1-6 aminoalkyl, =O or =S}

each L _ii is independently -(CH ₂ )-, -CHCH-, -C≡C-, -CH ₂ CH ₂ O- or _{-OCH 2} CH ₂ -, wherein at least one hydrogen in _{L ii is C 1 -6} alkyl, halo, C _1-6 haloalkyl, C _1-6 aminoalkyl},

s is an integer from 0 to 20;

q and r are each independently an integer from 0 to 10.

,

,

,

또는

이다{상기 L_T에서, L_Y는 단일결합, -(CH₂)_a-, -CO-, -O-, -NH- 또는 -NHCO-이고, a는 1 내지 6의 정수이며, TL은 화학식 1에 정의된 TL 모이어티를 나타내고,

는 화학식 L-2의 나머지 모이어티에 공유결합으로 연결됨을 나타냄}.In one embodiment of Formula L-2, L _T is a single bond, -CO(CH ₂ ) _a -,

,

,

,

or

Is {In L _T , L _Y is a single bond, -(CH ₂ ) _a -, -CO-, -O-, -NH- or -NHCO-, a is an integer of 1 to 6, TL is the formula represents the TL moiety defined in 1,

indicates that it is covalently linked to the remaining moieties of formula L-2}.

,

, -L_Y-(CH₂)_a-NH-, -L_Y-CH₂CH₂O-, -L_Y-CH₂CH₂NH- 또는

이다{상기 L_E에서, L_Y는 단일결합, -(CH₂)_a-, -CO-, -O-, -NH- 또는 -NHCO-이고, a는 1 내지 6의 정수이며, ELL은 화학식 1에 정의된 ELL 모이어티를 나타내고,

는 화학식 L-2의 나머지 모이어티에 공유결합으로 연결됨을 나타냄}.In one embodiment of Formula L-2, L _E is a single bond, -CH ₂ -, -NH-, -O-,

,

, -L _Y -(CH ₂ ) _a -NH-, -L _Y -CH ₂ CH ₂ O-, -L _Y -CH ₂ CH ₂ NH- or

Is {In the L _E , L _Y is a single bond, -(CH ₂ ) _a -, -CO-, -O-, -NH- or -NHCO-, a is an integer of 1 to 6, ELL is the formula represents the ELL moiety as defined in 1,

indicates that it is covalently linked to the remaining moieties of formula L-2}.

In one embodiment of Formula L-2 above, L _ii is —CH ₂ — or —CH ₂ CH ₂ O—.

In one embodiment of Formula L-2 above, q, s, and r are integers such that their sum is 1 to 30 such that the minimum number of atoms covalently linked between the TL and ELL moieties is 1 to 30.

According to a specific embodiment of the present invention, the linker has the structure of any one of formulas L-3-1 to L-3-26 shown in the table below. In the table below, TL and ELL represented by Formulas L-3-1 to L-3-26 represent the TL and ELL moieties defined in Formula 1 above, respectively, and k is a covalent bond between the TL and ELL moieties. It is an integer such that the minimum number of connected atoms is 1 to 30.

According to a specific embodiment of the present invention, the compound represented by Formula 1 is selected from the group consisting of compounds shown in Table 1 below.

[Table 1]

In the present invention, unless otherwise specified, "alkyl" may mean a straight or branched acyclic, cyclic, or saturated hydrocarbon to which they are bonded. For example, "C _1-6 alkyl" means alkyl containing 1 to 6 carbon atoms. Acyclic alkyl may include, as an example, methyl, ethyl, n -propyl, n -butyl, isopropyl, sec -butyl, isobutyl, or tert -butyl, etc., It is not limited thereto. Cyclic alkyl may be used interchangeably with “cycloalkyl” herein, and may include, as an example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl, but is limited thereto. doesn't happen

In the present invention, "alkoxy" may mean -(O-alkyl) as an alkyl ether group, where alkyl is as defined above. For example, "C _1-6 alkoxy" may _{mean alkoxy containing C 1-6} alkyl, that is, -(OC _1-6 alkyl). For example, alkoxy is methoxy ), ethoxy (ethoxy) n - propoxy (n -propoxy), iso-propoxy (isopropoxy), n - butoxy (n -butoxy), isobutoxy (isobutoxy), sec - butoxy (sec -butoxy) , or tert - butoxy, and the like, but (tert -butoxy), but is not limited thereto.

In the present invention, "halo" may be F, Cl, Br, or I.

In the present invention, "haloalkyl" may mean a straight or branched chain alkyl (hydrocarbon) having one or more halo-substituted carbon atoms as defined herein. Examples of such haloalkyl include, but are not limited to, methyl, ethyl, propyl, isopropyl, isobutyl or n-butyl independently substituted with one or more halogens, such as F, Cl, Br, or I. .

As used herein, "aminoalkyl" may mean a straight or branched chain alkyl (hydrocarbon) having a carbon atom substituted with amino-(NR'R"), where R' and R" are each independently hydrogen. , and _{may be selected from the group consisting of C 1-6} alkyl, wherein the selected R′ and R″ may each independently be substituted or unsubstituted.

In the present invention, "heterocycloalkyl" may mean a ring containing 1 to 5 heteroatoms selected from N, O and S as atoms forming the ring, and may be saturated or partially unsaturated. Here, when unsaturated, it may be referred to as a heterocycloalkene. Unless otherwise stated, a heterocycloalkyl may be a single ring or multiple rings such as a spiro ring, a bridged ring or a fused ring. In addition, "3 to 12 membered heterocycloalkyl" may mean a heterocycloalkyl containing 3 to 12 atoms forming a ring, for example, heterocycloalkyl is pyrrolidine, piperidine, Imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, piperidine, pyrimidine-2,4( 1H , 3H ) -Dione, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine- S -oxide, thiomorpholine- S , S -oxide, piperazine, pyran, pyridone, 3-pyrroline , Sy O-pyran, Piron, tetrahydrofuran, tetrahydro-thiophene, quinuclidine, tropane, 2-aza-Spy to [3.3] heptane, (1 R, 5 S) -3- azabicyclo [3.2. ] octane, (1 s, 4 s) -2- azabicyclo [2.2.2] octane include, or the like (1 R, 4 R) -2- oxa-5-azabicyclo [2.2.2] octane can, but is not limited thereto.

In the present invention, "arene" may mean an aromatic hydrocarbon ring. The arene may be a monocyclic arene or a polycyclic arene. The ring carbon number of arene may be 5 or more and 30 or less, 5 or more and 20 or less, or 5 or more and 15 or less. Examples of arenes include benzene, naphthalene, fluorene, anthracene, phenanthrene, bibenzene, terbenzene, quaterbenzene, quinkbenzene, sexbenzene, triphenylene, pyrene, benzofluoranthene, chrysene, etc. , but not limited to these. In the present specification, the residue obtained by removing one hydrogen atom from the "arene" is referred to as "aryl".

In the present invention, "heteroarene" may be a ring including at least one of O, N, P, Si, and S as a heterogeneous element. The number of ring carbon atoms of heteroarene may be 2 or more and 30 or less, or 2 or more and 20 or less. The hetero arene may be a monocyclic hetero arene or a polycyclic hetero arene. The polycyclic heteroarene may have, for example, a two- or three-ring structure. Examples of heteroarenes include thiophene, purine, pyrrole, pyrazole, imidazole, thiazole, oxazole, isothiazole, oxadiazole, triazole, pyridine, bipyridyl, triazine, acridyl, pyridazine , pyrazine, quinoline, quinazoline, quinoxaline, phenoxazine, phthalazine, pyrimidine, pyridopyrimidine, pyridopyrazine, pyrazinopyrazine, isoquinoline, indole, carbazole, imidazopyridazine, imidazopyridine , imidazopyrimidine, pyrazolopyrimidine, imidazopyrazine or pyrazolopyridine, N -arylcarbazole, N -heteroarylcarbazole, N -alkylcarbazole, benzoxazole, benzoimidazole, benzothiazole , benzocarbazole, benzothiophene, dibenzothiophene, thienothiophene, benzofuran, phenanthroline, isoxazole, oxadiazole, thiadiazole, benzothiazole, tetrazole, phenothiazine , dibenzosilol and dibenzofuran, but is not limited thereto. In one embodiment of the invention heteroarenes may also include bicyclic heterocyclo-arenes, including arene rings fused to heterocycloalkyl rings or heteroarenes fused to cycloalkyl rings. In the present specification, the residue obtained by removing one hydrogen atom from the "heteroarene" is referred to as "heteroaryl".

In the present invention, the term "enantiomer" means a compound of the present invention or a salt thereof having the same chemical formula or molecular formula but sterically different. Each of these optical isomers and mixtures thereof are also included within the scope of the present invention. Unless otherwise specified, a solid line bond (-) connected to an asymmetric carbon atom may include a wedge-shaped solid line bond or a wedge-shaped dotted line bond indicating the absolute arrangement of stereocenters.

In the present invention, the pharmaceutically acceptable salt refers to any organic or inorganic acid addition salt that is a concentration having an effective action that is relatively non-toxic and harmless to a patient, and that side effects due to the salt do not reduce the beneficial efficacy of the compound represented by the formula (1). means For example, the pharmaceutically acceptable salt may be hydrochloric acid, phosphoric acid, sulfuric acid, or nitric acid as an inorganic acid, and methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, Can be tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid or hydroiodic acid However, it is not limited thereto.

Method for producing protein kinase degradation inducing compound

In the present invention, the protein kinase degradation-inducing compound represented by the above formula (1), a stereoisomer or a pharmaceutically acceptable salt thereof, is a synthetic method known in the art of organic chemistry or modification and derivatization obvious to those skilled in the art. It can be prepared by a reaction such as the following Schemes 1 to 3 by the technique.

[Scheme 1]

[Scheme 2]

[Scheme 3]

In Schemes 1 to 3, TL, Linker and ELL are as defined above, and RG ¹ , RG ² , RG ^2a , RG ^2b , RG ³ , RG ^3a , RG ^3b and RG ⁴ are covalent bonds in the field of organic synthesis It is a moiety comprising a suitable reactive group capable of linking together the intermediate PROTAC compound represented by formula (1) through formation. The covalent bond formation is amide formation, ester formation, carbamate formation, urea formation, ether formation, amine formation, and various carbon-to-carbon single bonds, double bonds formation, and click chemistry according to specific reactive groups. may be formed, but is not limited thereto.

A variation of each step in the above scheme may include one or multiple synthetic steps. Isolation and purification of the product can be accomplished by standard procedures known to those skilled in the art of organic chemistry.

In the present invention, the preparation method of Example 64 is presented as an example of Scheme 1, and the preparation methods of Examples 1-52, 54-63, 65-66, and 69-73 are presented as an example of Scheme 2, As an example of 3, the manufacturing methods of Examples 53, 67, and 68 were presented, but the manufacturing method of the present invention is not limited thereto.

In the above scheme, the compounds represented by the chemical formulas A-1, B-1 and C-1, which are the reactants represented by Staurosporine and ELL, as a reactant represented by TL, are known in the organic chemistry field and the practice of the present invention. It can be easily synthesized by those skilled in the art with reference to examples and the like.

Use of protein kinase degradation inducing compounds

One embodiment of the present invention is a composition for inducing degradation of AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutagenesis comprising the compound represented by Formula 1, a stereoisomer or a pharmaceutically acceptable salt thereof. Formula 1 is as defined above.

The composition for inducing degradation of the present invention is ARK5, AURKA, AURKB, AURKC, CAMK1G, CAMK2A, CAMK2D, CAMKK1, CAMKK2, DYRK1A, EGFR (L858R/T790M), EGFR (T790M), EGFR (L858R)), EGFR (D858R)) EGFR(C797S), EGFR(Del19/C797S), EGFR(T790M/C797S), EGFR(Del19/T790M/C797S), EGFR(L858R/T790M/C797S), FLT3(D835H), FLT3(D835V835Y), FLT3(D835V835Y) ), FLT3 (ITD), FLT3 (ITD/D835V), FLT3 (ITD/F691L), FLT3 (K663Q), FLT3 (N841I), FLT3 (R834Q), GRK1, GRK7, IKK-epsilon, IRAK3, JAK2 (JH1domain- catalytic), JAK3 (JH1domain-catalytic), KIT (A829P), KIT (D816H), KIT (D816V), MAP4K2, MARK1, MLK1, MLK3, MST2, PDGFRB, PHKG1, PKN1, PKN2, PLK4, PRKK2 , QSK, RET (V804M), RSK2 (Kin.Dom.1-N-terminal), RSK4 (Kin.Dom.1-N-terminal), SIK2, SNARK, TBK1, TNK1, TRKA, TRKB, or YSK4 You may. For reference, a parenthesis '( )' among the protein kinases means a mutation.

In an embodiment of the present invention, the compound of the present invention comprises AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation, specifically, AURKA, FLT3 (ITD), EGFR (Del19, C797S, Del19/C797S, T790M/C797S, Del19/T790M/C797S), it was confirmed that it effectively induced the degradation of c-KIT (D816V). Accordingly, the composition comprising the compound represented by Formula 1 of the present invention, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof may be usefully used for inducing degradation of AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation. can

One embodiment of the present invention provides an AURKA, FLT3 mutant, EGFR mutation, or c-KIT mutation, specifically, AURKA, FLT3 ( ITD), EGFR (Del19, C797S, Del19/C797S, T790M/C797S, Del19/T790M/C797S), or c-KIT (D816V) is a pharmaceutical composition for the treatment or prevention of related diseases. Formula 1 is as defined above.

In the present invention, diseases related to AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation can be treated, alleviated, delayed, inhibited or prevented from inducing degradation or inhibiting activity of AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation. any disease or condition with

In one embodiment of the present invention, AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation, specifically, AURKA, FLT3(ITD), EGFR(Del19, C797S, Del19/C797S, T790M/C797S, Del19/T790M/ C797S), or c-KIT (D816V)-associated disease may be cancer or mastocytosis.

In the present invention, the cancer may include both blood cancer and solid cancer. When the cancer is a blood cancer, it may include both acute leukemia and chronic leukemia. In addition, when the cancer is acute leukemia, it may include acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL). In the case of acute myeloid leukemia (AML), it may be a gene mutation AML. Specifically, acute myeloid leukemia (AML) may be FLT3 mutation AML or c-KIT mutation AML, and more specifically acute myeloid leukemia (AML) may be FLT3 (ITD) mutation AML or c-KIT (D816V) mutation AML. .

In the present invention, the mastocytosis may be systemic mastocytosis (SM).

The pharmaceutical composition of the present invention may further include one or more pharmaceutically acceptable carriers in addition to the compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof for administration. These formulations may be prepared by referring to a conventional method or literature [Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA] used for formulation in the art, and may be formulated into various formulations according to each disease or component.

The pharmaceutical composition of the present invention may be administered orally or parenterally (eg, intravenously, subcutaneously, intraperitoneally or topically) according to a desired method, and the dosage may vary depending on the patient's weight, age, sex, and health condition. , diet, administration time, administration method, excretion rate, and the severity of the disease, etc., the range varies.

The pharmaceutical composition of the present invention may further include one or more active ingredients exhibiting the same or similar efficacy in addition to the compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

In addition, according to another embodiment of the present invention, the present invention provides AURKA, FLT3 mutation, EGFR mutation, AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation, specifically, AURKA, FLT3 (ITD), EGFR (Del19, C797S, Del19/C797S, T790M/C797S, Del19/T790M/C797S) or c-KIT (D816V). .

In another embodiment of the present invention, AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation by administering the compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof to a sample in vitro method to decompose. The sample may be a cell, a cell culture, a body fluid or tissue of a mammal including a human, but is not limited thereto.

In addition, according to another embodiment of the present invention, the present invention provides AURKA, FLT3 mutation, EGFR mutation by administering a compound represented by Formula 1, a stereoisomer, or a pharmaceutically acceptable salt thereof to mammals including humans. , or a method for treating a c-KIT mutation-related disease.

In addition, according to another embodiment of the present invention, the present invention provides a compound represented by Formula 1 for the treatment or prevention of AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation-related disease, a stereoisomer thereof, or a pharmaceutical thereof Provided are the uses of the commercially acceptable salts.

The compounds of the present invention contain AURKA, FLT3 mutation, EGFR mutation, or c-KIT mutation, specifically, AURKA, FLT3 (ITD), EGFR (Del19, C797S, Del19/C797S, T790M/C797S, Del19/T790M/C797S), or c-KIT (D816V) has an effect inducing degradation. Therefore, the compound of the present invention is useful for preventing or treating AURKA, FLT3 (ITD), EGFR (Del19, C797S, Del19/C797S, T790M/C797S, Del19/T790M/C797S), or c-KIT (D816V) related diseases. can be used

1 is a diagram showing the degradation results of AUKRA, FLT3 (ITD) of compound 5 of the present invention in the MV4-11 cell line.
2 is a diagram showing the degradation results of AUKRA of compound 5 of the present invention in Molt4 cell line.
3 is a diagram showing the degradation results of c-KIT (D816V) of compound 5 of the present invention in HMC1.2 cell line.
4 is a diagram showing the degradation results of AUKRA and FLT3 of compound 5 of the present invention in Molm14 cell line.
5 is a diagram showing the results of inhibition of PROTAC-induced degradation of AUKRA and FLT3 of compound 5 of the present invention after pretreatment with MLN4924 at a concentration of 1 uM in Molm14 cell line for 4 hours. The MLN4924 is an NEDD8-Activating Enzyme (NAE) E1 inhibitor, and NAE is an essential component of the NEDD8 conjugation pathway that modulates the subtype activity of ubiquitin ligase to convert and regulate upstream proteins of the proteasome.
6 is a diagram showing the results of inhibition of degradation of AUKRA and FLT3 of the reference material of the present invention (negative control of Compound 5) in Molm14 cell line.

Hereinafter, the configuration and effect of the present invention will be described in more detail through Examples and Experimental Examples. These Examples and Experimental Examples are only for illustrating the present invention, and the scope of the present invention is not limited by these Examples.

Example 1. Preparation of compound 1

Step 1: To tert-Butyl3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3- dioxoisoindolin-4-yl)amino) Preparation of oxy) ethoxy) propanoate

To 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (1.0 equiv), tert-butyl3-(2-(2-aminoethoxy) Toxy)propanoate (1.1 equiv.) and DIPEA (2.0 equiv.) were dissolved in NMP (0.3 M), followed by stirring at 110° C. for 3 hours. After the reaction material was cooled to room temperature, water was poured into it, and the mixture was extracted with ethyl acetate (x3). The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by column chromatography to obtain the target compound (yield: 44%). MS (ESI): m/z 490 [M+1] ⁺

Step 2: 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy ) production of propanoic acid

tert-Butyl3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino) prepared in step 1 Ethoxy) ethoxy) propanoate (1.0 equivalent) was dissolved in 1:2 (v/v) of TFA and CH ₂ Cl ₂ , followed by stirring at room temperature for 1 hour. After filtration of the reaction solution, the solvent was removed under reduced pressure to obtain the target compound without purification (yield: 100%). MS (ESI): m/z 434 [M+1] ⁺

Step 3: 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy )-N-((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa Preparation of -4b,9a,15-triaza-5,9-methanedibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)-N-methylpropanamide

3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy) prepared in step 2 Ethoxy) propanoic acid (1.0 equiv.) and Staurosporine (1.0 equiv.) were dissolved in DMF (0.2 M), HATU (1.0 equiv.) and DIPEA (4.0 equiv.) were added, followed by stirring at room temperature for 2 hours. After adding saturated NaHCO ₃ , it was extracted three times with ethyl acetate. The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by prep-HPLC to obtain the target compound as a yellow solid (yield: 20%). MS (ESI): m/z 883 [M+1] ⁺

Examples 2 to 8. Preparation of compounds 2 to 8

The compounds of Examples 2 to 8 according to the present invention were prepared in a manner similar to that of Example 1. The chemical structural formulas of each Example are summarized in Table 1, and the compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 2.

Example 9. Preparation of compound 9

Step 1: Preparation of 2-(2-(((benzyloxy)carbonyl)amino)ethoxy)ethyl 4-methylbenzenesulfonate

2-[N-(benzyloxycarbonyl)aminoethoxy]ethanol (1.0 equiv.) and TEA (1.5 equiv. _{) were dissolved in CH 2} Cl ₂ (0.2 M), followed by p-toluenesulfonylchloride (1.0 equiv.) was added and stirred at room temperature for 16 hours. After the reaction, the reaction solution was removed under reduced pressure, and water was added to the resultant, followed by extraction with ethyl acetate. The organic layer was _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by column chromatography to obtain the target compound as a white solid (yield: 60%). MS (ESI): m/z 394 [M+1] ⁺

Step 2: Preparation of tert-butyl 3-(2-(2-(((benzyloxy)carbonyl)amino)ethoxy)ethoxy)benzoate

After dissolving tert-butyl 3-hydroxybenzoate (1.5 eq.) and K ₂ CO ₃ (3.0 eq.) in MeCN (0.2 M), 2-(2-(((benzyloxy)carbonyl) prepared in step 1 )Amino)ethoxy)ethyl 4-methylbenzenesulfonate (1.0 equiv) was added and stirred at 80° C. for 12 h. After cooling to room temperature, the solvent was removed under reduced pressure. The resultant was dissolved in ethyl acetate and washed sequentially with water and brine. The organic layer _{was dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by column chromatography to obtain the target compound (yield: 85%). MS (ESI): m/z 416 [M+1] ⁺

Step 3: Preparation of tert-butyl 3-(2-(2-aminoethoxy)ethoxy)benzoate

After dissolving tert-butyl 3-(2-(2-(((benzyloxy)carbonyl)amino)ethoxy)ethoxy)benzoate (1.0 equivalent) prepared in step 2 in methanol (0.2 M), Pd /C (0.1 equiv) was added. The mixture was stirred at room temperature under hydrogen gas for 12 hours. The reaction product was filtered through celite, and washed with ethyl acetate. The solvent was removed from the filtrate under reduced pressure to obtain the target compound without purification (yield: 92%). MS (ESI): m/z 282 [M+1] ⁺

Step 4: tert-Butyl 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy ) Preparation of ethoxy) benzoate

tert-Butyl 3-(2-(2-aminoethoxy)ethoxy)benzoate (1.0 equiv) prepared in step 3, 2-(2,6-dioxopiperidin-3-yl)-4-fluoro After dissolving loisoindoline-1,3-dione (1.0 equiv.) and DIPEA (2.0 equiv.) in NMP (0.3 M), the mixture was stirred at 110° C. for 3 hours. After the reaction material was cooled to room temperature, water was poured therein, and the mixture was extracted with ethyl acetate (x3). The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by column chromatography to obtain the target compound (yield: 38%). MS (ESI): m/z 538 [M+1] ⁺

Step 5: 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy ) Preparation of benzoic acid

tert-Butyl 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino) prepared in step 4 Ethoxy)ethoxy)benzoate (1.0 equivalent) was dissolved in 1:2 (v/v) TFA and CH ₂ Cl ₂ , followed by stirring at room temperature for 1 hour. After filtration of the reaction solution, the solvent was removed under reduced pressure to obtain the target compound without purification (yield: 100%). MS (ESI): m/z 482 [M+1] ⁺

Step 6: 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy )-N-((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa Preparation of -4b,9a,15-triaza-5,9-methanedibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)-N-methylbenzamide

3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy) prepared in step 5 After dissolving ethoxy)benzoic acid (1.0 equiv.) and Staurosporine (1.0 equiv.) in DMF (0.2 M), HATU (1.0 equiv.) and DIPEA (4.0 equiv.) were added, followed by stirring at room temperature for 2 hours. After adding saturated NaHCO ₃ , it was extracted three times with ethyl acetate. The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by prep-HPLC to obtain the target compound (yield: 30%).

MS (ESI): m/z 930 [M+1] ⁺

Examples 10 to 12. Preparation of compounds 10 to 12

Examples 10 to 12 compounds according to the present invention were prepared in a manner similar to that of Example 9. The chemical structural formulas of each Example are summarized in Table 1, and the compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 2.

Example 13. Preparation of compound 13

Step 1: Preparation of dibenzyl 3-((2-(2-hydroxyethoxy)ethyl)amino)phthalate

After dissolving dibenzyl 3-fluorophthalate (1.0 equiv.), 2-(2-aminoethoxy)ethan-1-ol (1.0 equiv.), and DIPEA (2.0 equiv.) in NMP (0.3 M), stirred for hours. After the reaction material was cooled to room temperature, water was poured into it, and the mixture was extracted with ethyl acetate (x3). The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by column chromatography to obtain the target compound (yield: 41%).

MS (ESI): m/z 450 [M+1] ⁺

Step 2: Preparation of dibenzyl 3-((2-(2-hydroxyethoxy)ethyl)amino)phthalate

After dissolving dibenzyl 3-((2-(2-hydroxyethoxy)ethyl)amino)phthalate (1.0 equiv.) and TEA (1.5 equiv.) prepared in step 1 in CH ₂ Cl ₂ (0.2 M), p -Toluenesulfonyl chloride (1.0 equivalent) was added and stirred at room temperature for 4 hours. The reaction solution was removed under reduced pressure, and water was added to the resultant, followed by extraction with ethyl acetate. The organic layer was _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by column chromatography to obtain the target compound (yield: 74%). MS (ESI): m/z 604 [M+1] ⁺

Step 3: Preparation of dibenzyl 3-((2-(2-(4-(tert-butoxycarbonyl)phenoxy)ethoxy)ethyl)amino)phthalate

After dissolving tert-butyl 4-hydroxybenzoate (1.5 eq.) and K ₂ CO ₃ (3.0 eq.) in MeCN (0.2 M), dibenzyl 3-((2-(2-hydroxyl) prepared in step 2 Ethoxy)ethyl)amino)phthalate (1.0 eq.) was added and stirred at 70° C. for 16 h. After cooling to room temperature, the solvent was removed under reduced pressure. The resultant was dissolved in ethyl acetate and washed sequentially with water and brine. The organic layer _{was dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by column chromatography to obtain the target compound (yield: 80%). MS (ESI): m/z 626 [M+1] ⁺

Step 4: Preparation of 3-((2-(2-(4-(tert-butoxycarbonyl)phenoxy)ethoxy)ethyl)amino)phthalic acid

Dibenzyl 3-((2-(2-(4-(tert-butoxycarbonyl)phenoxy)ethoxy)ethyl)amino)phthalate (1.0 equivalent) prepared in step 3 was dissolved in methanol (0.2 M) Then Pd/C (0.1 eq) was added. The mixture was stirred at room temperature under hydrogen gas for 1 hour. The reaction result was filtered through Celite, and washed with ethyl acetate. The solvent was removed from the filtrate under reduced pressure to obtain the target compound without purification (yield: 84%). MS (ESI): m/z 446 [M+1] ⁺

Step 5: tert-Butyl4-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy ) Preparation of ethoxy) benzoate

3-((2-(2-(4-(tert-butoxycarbonyl)phenoxy)ethoxy)ethyl)amino)phthalic acid (1.0 eq.) and 3-aminopiperidine-2 prepared in step 4, After adding 6-dione hydrochloride (1.0 equivalent) to pyridine, the mixture was stirred at 110° C. for 2 hours. The solvent was removed from the reaction product under reduced pressure. The concentrated material was purified using prep-HPLC to obtain the target compound (yield: 48%).

MS (ESI): m/z 538 [M+1] ⁺

Step 6: 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy ) Preparation of benzoic acid

tert-Butyl4-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino) prepared in step 5 4N HCl/dioxane was added to ethoxy)ethoxy)benzoate (1.0 equivalent) and stirred at room temperature for 1 hour. After filtration of the reaction solution, the solvent was removed under reduced pressure to obtain the target compound without purification (yield: 100%).

MS (ESI): m/z 482 [M+1] ⁺

Step 7: 4-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy )-N-((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa Preparation of -4b,9a,15-triaza-5,9-methanedibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)-N-methylbenzamide

3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy) prepared in step 6 After dissolving ethoxy)benzoic acid (1.0 equiv.) in CH ₂ Cl ₂ (0.2 M), SOCl ₂ (1.2 equiv.) was added at room temperature, and the temperature was raised to 45° C. and stirred for 1 hour. After the reaction, the solvent was removed under reduced pressure to remove 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino ) ethoxy) ethoxy) benzoyl chloride was obtained.

Staurosporine (1.0 equiv.) and TEA (3.0 equiv. _{) were dissolved in CH 2} Cl ₂ (0.4 M), and then 3-(2-(2-((2-(2,6-dioxopiperidine-3- A solution of yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)benzoyl chloride in CH ₂ Cl ₂ (0.4 M) was added and stirred at room temperature for 16 hours. After adding saturated NaHCO ₃ , it was extracted three times with ethyl acetate. The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified using prep-HPLC to obtain the target compound as a yellow solid (yield: 12%). MS (ESI): m/z 930 [M+1] ⁺

Example 14. Preparation of compound 14

Step 1: Preparation of 3-oxo-1-phenyl-2,7,10-trioxa-4-azadodecan-12-yl-4-methylbenzenesulfonate

Benzyl(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)carbamate (1.0 equiv.) and TEA (1.5 equiv. _{) were dissolved in CH 2} Cl ₂ (0.2 M), followed by p-toluenesulfonyl Chloride (1.0 equiv) was added and stirred at room temperature for 16 hours. The reaction solution was removed under reduced pressure, and water was added to the resultant, followed by extraction with ethyl acetate. The organic layer was _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by column chromatography to obtain the target compound as a white solid (yield: 90%). MS (ESI): m/z 438 [M+1] ⁺

Step 2: Preparation of tert-butyl4-((3-oxo-1-phenyl-2,7,10-trioxa-4-azadodecan-12-yl)oxy)benzoate

After dissolving tert-butyl4-hydroxybenzoate (1.5 eq.) and K ₂ CO ₃ (3.0 eq.) in MeCN (0.2 M), 3-oxo-1-phenyl-2,7,10 prepared in step 1 -Trioxa-4-azadodecan-12-yl-4-methylbenzenesulfonate (1.0 equiv) was added and stirred at 80° C. for 18 hours. After cooling to room temperature, the solvent was removed under reduced pressure. The resultant was dissolved in ethyl acetate and washed sequentially with water and brine. The organic layer _{was dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by column chromatography to obtain the target compound. (Yield: 83%) MS (ESI): m/z 460 [M+1] ⁺

Step 3: Preparation of tert-butyl4-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)benzoate

tert-Butyl4-((3-oxo-1-phenyl-2,7,10-trioxa-4-azadodecan-12-yl)oxy)benzoate (1.0 eq.) prepared in step 2 was mixed with methanol ( 0.2 M), then Pd/C (0.1 eq) was added. A balloon filled with hydrogen gas was inserted and stirred at room temperature for 3 hours. The reaction product was filtered through celite, and washed with ethyl acetate. The solvent was removed from the filtrate under reduced pressure to obtain the target compound without purification (yield: 87%). MS (ESI): m/z 326 [M+1] ⁺

Step 4: tert-Butyl4-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino) Preparation of ) ethoxy) ethoxy) ethoxy) benzoate

tert-Butyl4-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)benzoate (1.0 equiv) prepared in step 3, 2-(2,6-dioxopiperidine-3- yl)-4-fluoroisoindoline-1,3-dione (1.0 equiv.) and DIPEA (2.0 equiv.) were dissolved in NMP (0.3 M), followed by stirring at 110° C. for 3 hours. After the reaction material was cooled to room temperature, water was poured into it, and the mixture was extracted with ethyl acetate (x3). The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by column chromatography to obtain the target compound (yield: 27%). MS (ESI): m/z 582 [M+1] ⁺

Step 5: 4-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy ) Preparation of ethoxy) ethoxy) benzoic acid

tert-Butyl4-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl) prepared in step 4) ) Amino) ethoxy) ethoxy) ethoxy) benzoate (1.0 equivalent) was added to 4N HCl/dioxane and stirred at room temperature for 1 hour. After filtration of the reaction solution, the solvent was removed under reduced pressure to obtain the target compound without purification (yield: 100%). MS (ESI): m/z 526 [M+1] ⁺

Step 6: 4-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy )ethoxy)ethoxy)-N-((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H , 14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)-N -Preparation of methylbenzamide

4-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino) prepared in step 5 After dissolving ethoxy)ethoxy)ethoxy)benzoic acid (1.0 equiv.) in CH ₂ Cl ₂ (0.2 M), SOCl ₂ (1.2 equiv.) was added at room temperature, and the temperature was raised to 45° C. and stirred for 1 hour. After the reaction, the solvent was removed under reduced pressure to remove 4-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4- yl) amino) ethoxy) ethoxy) ethoxy) benzoyl chloride was obtained.

Staurosporine (1.0 equiv.) and TEA (3.0 equiv. _{) were dissolved in CH 2} Cl ₂ (0.4 M), and then 4-(2-(2-(2-((2-(2,6-dioxopiperidine) -3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)benzoylchloride in CH ₂ Cl ₂ (0.4 M) was added to a solution of 16 stirred for hours. After adding saturated NaHCO ₃ , it was extracted three times with ethyl acetate. The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified using prep-HPLC to obtain the target compound as a yellow solid (yield: 12%). MS (ESI): m/z 974 [M+1] ⁺

Examples 15 to 17. Preparation of compounds 15 to 17

Examples 15 to 17 compounds according to the present invention were prepared in a manner similar to that of Example 14. The chemical structural formulas of each Example are summarized in Table 1, and the compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 2.

Example 18. Preparation of compound 18

Step 1: tert-Butyl 3-(2-(3-((((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazole-5-) Preparation of yl)benzyl)carbamoyl)pyrrolidin))-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy)propanoate

3-(2-(3-(tert-butoxy)-3-oxopropoxy)ethoxy)propanoic acid (1.0 eq) and (2S,4R)-1-((S)-2-amino-3, 3-Dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (1.0 equiv.) was dissolved in DMF (0.2 M) After dissolving, HATU (1.0 equivalent) and DIPEA (4.0 equivalent) were added and stirred at room temperature for 16 hours. After adding saturated NaHCO ₃ , it was extracted three times with ethyl acetate. The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by prep-HPLC to obtain the target compound (yield: 42%). MS (ESI): m/z 675 [M+1] ⁺

Step 2: 3-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl) Preparation of carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy)propanoic acid

tert-Butyl3-(2-(3-((((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazole-(4-(4-methylthiazole-) prepared in step 1) 5-yl)benzyl)carbamoyl)pyrrolidin))-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy)propanoate (1.0 equivalent) was dissolved in 1:2 (v/v) of TFA and CH ₂ Cl ₂ , followed by stirring at room temperature for 1 hour. After filtration of the reaction solution, the solvent was removed under reduced pressure to obtain the target compound without purification (yield: 100%).

MS (ESI): m/z 619 [M+1] ⁺

Step 3: (2S,4R)-1-((S)-14-(tert-butyl)-2-((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo- 6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclo Penta[e]-as-indasen-7-yl)-3,12-dioxo-6,9-dioxa-2,13-diazapentadecane-15-oil)-4-hydroxy-N- Preparation of (4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

3-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl) prepared in step 2) Benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy)propanoic acid (1.0 eq) and Staurosporine ( 1.0 eq.) was dissolved in DMF (0.2 M), HATU (1.0 eq.) and DIPEA (4.0 eq.) were added and stirred at room temperature for 16 hours. After adding saturated NaHCO ₃ , it was extracted three times with ethyl acetate. The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by prep-HPLC to obtain the target compound (yield: 44%). MS (ESI): m/z 1068 [M+1] ⁺

Examples 19 to 21. Preparation of compounds 19 to 21

The compounds of Examples 19 to 21 according to the present invention were prepared in a manner similar to that of Example 18. The chemical structural formulas of each Example are summarized in Table 1, and the compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 2.

Example 22. Preparation of compound 22

Step 1: Preparation of methyl 3-(2-(2-(tosyloxy)ethoxy)ethoxy)propanoate

After dissolving methyl 3-(2-(2-hydroxyethoxy)ethoxy)propanoate (1.0 equiv.) and TEA (1.5 equiv.) in CH ₂ Cl ₂ (0.2 M), p-toluenesulfonylchloride ( 1.0 equivalent) and stirred at room temperature for 16 hours. The reaction solution was removed under reduced pressure, and water was added to the resultant, followed by extraction with ethyl acetate. The organic layer was _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by column chromatography to obtain the target compound (yield: 65%).

MS (ESI): m/z 347 [M+1] ⁺

Step 2: Preparation of tert-butyl 3-(2-(2-(3-methoxy-3-oxopropoxy)ethoxy)ethoxy)benzoate

After dissolving tert-butyl 3-hydroxybenzoate (1.5 eq.) and K ₂ CO ₃ (3.0 eq.) in MeCN (0.2 M), methyl 3-(2-(2-(tosyloxy) prepared in step 1) Ethoxy)ethoxy)propanoate (1.0 eq.) was added and stirred at 70° C. for 12 hours. After cooling to room temperature, the solvent was removed under reduced pressure. The resultant was dissolved in ethyl acetate and washed sequentially with water and brine. The organic layer _{was dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by column chromatography to obtain the target compound (yield: 66%). MS (ESI): m/z 369 [M+1] ⁺

Step 3: Preparation of 3-(2-(2-(3-(tert-butoxycarbonyl)phenoxy)ethoxy)ethoxy)propanoic acid

tert-Butyl 3-(2-(2-(3-methoxy-3-oxopropoxy)ethoxy)ethoxy)benzoate (1.0 equiv.) prepared in step 2 and LiOH (1.5 equiv.) were mixed with MeOH/H _{After dissolving in 2} O (1:1), the mixture was stirred at room temperature for 1 hour. The solvent was concentrated under reduced pressure, and the resulting material was purified using prep-HPLC to obtain the target compound (yield: 70%). MS (ESI): m/z 355 [M+1] ⁺

Step 4: tert-Butyl3-(2-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazole- 5-yl)benzyl)))) carbamoyl) pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) amino)-3-oxopropoxy) ethoxy) ethoxy ) Preparation of benzoate

3-(2-(2-(3-(tert-butoxycarbonyl)phenoxy)ethoxy)ethoxy)propanoic acid (1.0 eq.) and (2S,4R)-1-(( S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (1.0 equivalent) was dissolved in DMF (0.2 M), HATU (1.0 equivalent) and DIPEA (4.0 equivalent) were added and stirred at room temperature for 16 hours. After adding saturated NaHCO ₃ , it was extracted three times with ethyl acetate. The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by prep-HPLC to obtain the target compound (yield: 51%).

MS (ESI): m/z 767 [M+1] ⁺

Step 5: 3-(2-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl) )benzyl)carbamoyl)pyrrolidin)))-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy)ethoxy)benzoic acid Produce

tert-Butyl3-(2-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthio) prepared in step 4) azol-5-yl)benzyl))))carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy) Ethoxy) benzoate (1.0 equivalent) was dissolved in 1:2 (v/v) TFA and CH ₂ Cl ₂ , followed by stirring at room temperature for 1 hour. After filtration of the reaction solution, the solvent was removed under reduced pressure to obtain the target compound without purification (yield: 100%). MS (ESI): m/z 711 [M+1] ⁺

Step 6: (2S,4R)-4-hydroxy-1-((S)-2-(3-(2-(2-(3-(((5S,6R,7R,9R)-6-methyl) oxy-5-methyl))-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9-methanodi Benzo[b,h]cyclonona[jkl]cyclopenta[e]-indacenda-7-yl)(methyl)carbamoyl)phenoxy)ethoxy)ethoxy)propanamido)-3,3-dimethylbuta Preparation of noyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

3-(2-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazole-5) prepared in step 5) -yl)benzyl)carbamoyl)pyrrolidin)))-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy)ethoxy) Benzoic acid (1.0 equiv.) and staurosporine (1.0 equiv.) were dissolved in DMF (0.2 M), HATU (1.0 equiv.) and DIPEA (4.0 equiv.) were added, followed by stirring at room temperature for 16 hours. After adding saturated NaHCO ₃ , it was extracted three times with ethyl acetate. The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified using prep-HPLC to obtain the target compound as a yellow solid (yield: 40%). MS (ESI): m/z 1160 [M+1] ⁺

Examples 23 to 25. Preparation of compounds 23 to 25

Examples 23 to 25 compounds according to the present invention were prepared in a manner similar to Example 22. The chemical structural formulas of each Example are summarized in Table 1, and the compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 2.

Example 26. Preparation of compound 26

Step 1: Preparation of tert-butyl 4-(2-(2-(3-methoxy-3-oxopropoxy)ethoxy)ethoxy)benzoate

tert-Butyl 4-hydroxybenzoate (1.5 equiv.) and K ₂ CO ₃ (3.0 equiv.) were dissolved in MeCN (0.2 M), followed by methyl 3-(2-(2-(tosyloxy)ethoxy)ethoxy) ) propanoate (1.0 eq.) was added and stirred at 60° C. for 16 hours. After cooling to room temperature, the solvent was removed under reduced pressure. The resultant was dissolved in ethyl acetate and washed sequentially with water and brine. The organic layer _{was dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by column chromatography to obtain the target compound (yield: 78%). MS (ESI): m/z 369 [M+1] ⁺

Step 2: Preparation of 3-(2-(2-(4-(tert-butoxycarbonyl)phenoxy)ethoxy)ethoxy)propanoic acid

tert-Butyl 4-(2-(2-(3-methoxy-3-oxopropoxy)ethoxy)ethoxy)benzoate (1.0 equiv.) prepared in step 1 and LiOH (1.5 equiv.) were mixed with MeOH/H _{After dissolving in 2} O (1:1), the mixture was stirred at room temperature for 30 minutes. The solvent was concentrated under reduced pressure, and the resulting material was purified using prep-HPLC to obtain the target compound as a white solid (yield: 55%). MS (ESI): m/z 355 [M+1] ⁺

Step 3: tert-Butyl4-(2-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazole- 5-yl)benzyl)))) carbamoyl) pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) amino)-3-oxopropoxy) ethoxy) ethoxy ) Preparation of benzoate

3-(2-(2-(4-(tert-butoxycarbonyl)phenoxy)ethoxy)ethoxy)propanoic acid (1.0 eq.) and (2S,4R)-1-(( S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (1.0 equivalent) was dissolved in DMF (0.2 M), HATU (1.0 equivalent) and DIPEA (4.0 equivalent) were added and stirred at room temperature for 16 hours. After adding saturated NaHCO ₃ , it was extracted three times with ethyl acetate. The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by prep-HPLC to obtain the target compound (yield: 55%).

MS (ESI): m/z 767 [M+1] ⁺

Step 4: 4-(2-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl) )benzyl)carbamoyl)pyrrolidin)))-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy)ethoxy)benzoic acid Produce

tert-Butyl4-(2-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthio) prepared in step 3) azol-5-yl)benzyl))))carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy) Ethoxy) benzoate (1.0 equivalent) was dissolved in 1:2 (v/v) TFA and CH ₂ Cl ₂ , followed by stirring at room temperature for 1 hour. After filtration of the reaction solution, the solvent was removed under reduced pressure to obtain the target compound without purification (yield: 100%). MS (ESI): m/z 711 [M+1] ⁺

Step 5: (2S,4R)-4-hydroxy-1-((S)-2-(3-(2-(2-(4-(((5S,6R,7R,9R)-6-methyl) oxy-5-methyl))-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9-methanodi Benzo[b,h]cyclonona[jkl]cyclopenta[e]-indacenda-7-yl)(methyl)carbamoyl)phenoxy)ethoxy)ethoxy)propanamido)-3,3-dimethylbuta Preparation of noyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

4-(2-(2-(3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazole-5) prepared in step 4) -yl)benzyl)carbamoyl)pyrrolidin)))-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethoxy)ethoxy) Benzoic acid (1.0 equiv.) and Staurosporine (1.0 equiv.) were dissolved in DMF (0.2 M), HATU (1.0 equiv.) and DIPEA (4.0 equiv.) were added, followed by stirring at room temperature for 16 hours. After adding saturated NaHCO ₃ , it was extracted three times with ethyl acetate. The collected organic layers were washed with brine, _{dried over Na 2} SO ₄ , filtered, and the solvent was concentrated under reduced pressure. The concentrated material was purified by prep-HPLC to obtain the target compound (yield: 13%).

MS (ESI): m/z 1160 [M+1] ⁺

Examples 27 to 29. Preparation of compounds 27 to 29

Examples 27 to 29 compounds according to the present invention were prepared in a manner similar to that of Example 26. The chemical structural formulas of each Example are summarized in Table 1, and the compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 2.

[Table 2]

Examples 30 to 33. Preparation of compounds 30 to 33

The compounds of Examples 30 to 33 according to the present invention were prepared in a manner similar to that of Example 1. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

Example 34. Preparation of compound 34

Step 1: Preparation of tert-butyl4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butanoate

2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (1.0 equiv.), tert-butyl4-aminobutanoate (1.1 equiv) was dissolved in DMF After dissolution, DIPEA (2.0 equiv.) was added and the mixture was ^{stirred at 80 °} C for 16 h. LC-MS was used to confirm the product. After adding water, the reaction mixture was extracted 3 times with EA. The _{remaining water was removed using Na 2} SO ₄ , concentrated and purified by MPLC to obtain the target compound. (Yield: 89%) MS (ESI): m/z 360 [M+1] ⁺

Step 2: Preparation of 4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butanoic acid

tert-Butyl4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butanoate prepared in step 1 was mixed with 4.0 M After dissolving in HCl/Dioxane (1 mL), the mixture was stirred at room temperature for 1 hour. After confirming the product using LC-MS, the product was concentrated, and the next reaction proceeded without further purification. MS (ESI): m/z 360 [M+1] ⁺

Step 3: 4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-N-((5S,6R,7R, 9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5, Preparation of 9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)-N-methylbutanamide

4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butanoic acid (1.0 equiv) prepared in step 2, HATU ( 3.0 equiv.) was dissolved in DMF, and Staurosporine (1.0 equiv.) and DIPEA (2.0 equiv.) were also dissolved in DMF and stirred for 30 minutes each. The two reaction mixtures were mixed and stirred at room temperature for 1 hour. The product was confirmed by LC-MS and purified by prep-HPLC to obtain the target compound. (two-step yield: 6%) MS (ESI): m/z 808 [M+1] ⁺

Examples 35 to 40. Preparation of compounds 35 to 40

The compounds of Examples 35 to 40 according to the present invention were prepared in a manner similar to that of Example 34. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

Example 41. Preparation of compound 41

Step 1: Preparation of tert-butyl(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)glycinate

2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (1.0 eq.) was dissolved in DMSO (0.1 M), followed by tert-butylglycinate ( 1.0 equiv) and DIPEA (2.0 equiv) added to 90 The mixture was stirred at °C for 16 hours. After the reaction was cooled to room temperature, water and brine were added, and the organic material was extracted with EtOAc. The collected organic layers were concentrated after removing the remaining water using _{Na 2} SO _{4 .} The reaction mixture was purified by MPLC. The title compound was obtained as a yellow solid. (Yield: 30%)

MS (ESI): m/z 332 [M-(t-Bu)+1] ⁺

Step 2: Preparation of (2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)glycine

tert-butyl(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)glycinate (1.0 equiv.) prepared in step 1 was mixed with DCM (0.4 After dissolving in M), TFA (30 equivalents) was added and stirred at room temperature for 30 minutes. The reaction mixture was concentrated and purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 33%) MS (ESI): m/z 332 [M+1] ⁺

Step 3: tert-Butyl 1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-2-oxo-6,9 , Preparation of 12-trioxa-3-azapentadecane-15-oate

After dissolving (2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)glycine prepared in step 2 in DMF (0.05 M), HATU (2.0 equivalent) and DIPEA (3.0 equivalents) were added and stirred at room temperature for 30 minutes. To the reaction mixture was added tert-butyl3-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)propanoate (1.1 equivalents) and stirred at room temperature for 1 hour. Water and brine were added to the reaction mixture, and organics were extracted with EtOAc. The collected organic layers were concentrated after removing the remaining water using _{Na 2} SO _{4 .} The obtained target compound as a brown liquid was used in the next reaction without further purification. (Yield: 100%, MS (ESI): m/z 535 [M-(t-Bu)+1] ⁺ )

Step 4: 1-(2-(2-6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-2-oxa-6,9,12-trioxa- Preparation of 3-azapentadecane-15-oic acid

tert-Butyl1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-2-oxo-6 prepared in step 3 ,9,12-trioxa-3-azapentadecane-15-oate (1.0 equiv.) was dissolved in DCM (0.1 M), TFA (30 equiv) was added and stirred for 1 hour. The reaction mixture was concentrated and purified by prep-HPLC to obtain the target compound as a yellow solid. (Yield: 12%) MS (ESI): m/z 535 [M+1] ⁺

Step 5: 3-(2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino) )acetamido)ethoxy)ethoxy)ethoxy)-N-((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15 , 16-hexahydro-5H, 14H-17-oxa-4b, 9a, 15-triaza-5,9-methanodibenzo [b, h] cyclonona [jkl] cyclopenta [e] -as-indacene Preparation of -7-yl)-N-methylpropanamide

1-(2-(2-6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-2-oxa-6,9,12-tri prepared in step 4 After oxa-3-azapentadecane-15-oic acid (1.0 equiv.) was dissolved in DMF (0.05 M), HATU (3.0 equiv.) and DIPEA (2.0 equiv.) were added, followed by stirring at room temperature for 30 minutes. Staurosporine (1.0 equivalent) was added to the reaction mixture and stirred for 1 hour. Water and brine were added to the reaction mixture, and organics were extracted with EtOAc. The collected organic layers were concentrated after removing the remaining water using _{Na 2} SO _{4 .} The reaction mixture was purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 16%) MS (ESI): m/z 984 [M+1] ⁺

Example 42. Preparation of compound 42

The compound of Example 42 according to the present invention was prepared in a similar manner to Example 41. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

Example 43. Preparation of compound 43

Step 1: Preparation of tert-butyl2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetate

2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (1.0 equiv), tert-butyl2-bromoacetate (1.2 equiv), K ₂ CO ₃ (1.5 equivalents) was dissolved in DMF (0.2 M) and stirred at room temperature for 2 hours. Water and brine were added, and organic matter was extracted with EtOAc. The collected organic layer was concentrated after removing the remaining water using _{Na 2} SO _{4 .} The reaction mixture was purified through MPLC (DCM: MeOH) to give the title compound as a yellow solid. (Yield: 96%) MS (ESI): m/z [M-(t-Bu)+1] ⁺

Step 2: Preparation of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetic acid

tert-Butyl2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetate (1.0 equiv.) prepared in step 1 was After dissolving in TFA (0.1M), the mixture was stirred at room temperature for 1 hour. After concentrating the reaction mixture, it was solidified using ether, and the target compound as a yellow solid obtained was used in the next reaction without further purification. (Yield: 94%) MS (ESI): m/z 333 [M+1] ⁺

Step 3: tert-Butyl1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-2-oxo-6,9 , Preparation of 12-trioxa-3-azapentadecane-15-oate

2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetic acid (1.0 equiv) prepared in step 2, tert-butyl 3-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)propanoate (1.1 equiv.), HATU (2.0 equiv.), and DIPEA (3.0 equiv.) were dissolved in DMF (0.05 M) at room temperature stirred for 1 hour. Water and brine were added, and organic matter was extracted with EtOAc. The collected organic layer was concentrated after removing the remaining water using _{Na 2} SO _{4 .} The obtained target compound as a yellow solid was used in the next reaction without further purification. (Yield: 100%) MS (ESI): m/z 536 [M-(t-Bu)+1] ⁺

Step 4: 1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-2-oxo-6,9,12- Preparation of trioxa-3-azapentadecane-15-oic acid

tert-Butyl1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-2-oxo-6 prepared in step 3 ,9,12-trioxa-3-azapentadecane-15-oate (1.0 equivalent) was dissolved in 4 N HCl/1,4-Dioxane (0.1 M) and stirred at room temperature for 1 hour. The reaction mixture was concentrated and purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 20%) MS (ESI): m/z 536 [M+1] ⁺

Step 5: 3-(2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy )acetamido)ethoxy)ethoxy)ethoxy)-N-((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15 , 16-hexahydro-5H, 14H-17-oxa-4b, 9a, 15-triaza-5,9-methanodibenzo [b, h] cyclonona [jkl] cyclopenta [e] -as-indacene Preparation of -7-yl)-N-methylpropanamide

1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-2-oxo-6,9 prepared in step 4, 12-trioxa-3-azapentadecane-15-oic acid (1.0 equiv.), Staurosporine (1.1 equiv.), HATU (2.0 equiv.), and DIPEA (3.0 equiv.) were dissolved in DMF (0.05 M) and then at room temperature for 1 hour stirred for a while. Water and brine were added, and organic matter was extracted with EtOAc. The collected organic layers were concentrated after removing the remaining water using _{Na 2 SO4.} The reaction mixture was purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 58%) MS (ESI): m/z 984 [M+1] ⁺

Example 44. Preparation of compound 44

The compound of Example 44 according to the present invention was prepared in a manner similar to that of Example 43. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

Example 45. Preparation of compound 45

Step 1: Preparation of 2-(2,6-dioxopiperidin-3-yl)-4-(3-hydroxyprop-1-yn-1-yl)isoindoline-1,3-dione

4-bromo-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (1.0 equiv), prop-2-yn-1-ol (2.0 equiv) DMF: a the reaction mixture was 1 minutes N ₂ degassing, while maintaining the 80 ℃ CuI (0.2 eq), Pd (PPh ₃₎ Cl ₂ (0.1 eq.) dissolved in (1 ratio, 0.1 M 1) TEA and stirred for 16 hours. After confirming that the reaction was complete through LC-MS, the mixture was concentrated under reduced pressure to obtain the target compound as a brown oil through MPLC. (Yield: 78%) MS (ESI): m/z 313 [M+1] ⁺

Step 2: tert-Butyl16-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-4,7,10,13-tetraoxa Preparation of hexadec-15-inoate

2-(2,6-dioxopiperidin-3-yl)-4-(3-hydroxyprop-1-yn-1-yl)isoindoline-1,3-dione prepared in step 1 ( 1.0 equiv.) in DMF (0.1 M) in a reaction mixture of tert-butyl3-(2-(2-(2-iodoethoxy)ethoxy)ethoxy)propanoate (1.0 equiv.), Cs ₂ CO ₃ (2.0 equiv) was added and stirred at 40° C. for 30 min. After confirming that the reaction was completed through LC-MS, distilled water was added, the organic material was extracted with EA, the organic layer was washed with brine, the remaining water was removed with _{Na 2} SO _{4 , filtered, and concentrated.} The target compound was obtained without further purification. MS (ESI): m/z 517 [M-(t-Bu)+1] ⁺

Step 3: 16-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-4,7,10,13-tetraoxahexadec- Preparation of 15-inoic acid

tert-Butyl 16-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-4,7,10,13- prepared in step 2 Tetraoxahexadec-15-inoate (1.0 equivalent) was dissolved in 4 M HCl/1,4-Dioxane (0.1 M) and stirred at room temperature for 1 hour. The reaction mixture was concentrated and purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 23%) MS (ESI): m/z 517 [M+1] ⁺

Step 4: 16-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-N-((5S,6R,7R,9R)- 6-Methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9-meta of nodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)-N-methyl-4,7,10,13-tetraoxahexadec-15-inamide Produce

16-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-4,7,10,13-tetraoxahexa prepared in step 3 DIPEA (3.0 equiv.) and HATU (2.0 equiv.) were added to the reaction mixture in which Dec-15-inoic acid (1.0 equiv.) was dissolved in DMF (0.1 M), followed by stirring at room temperature for 30 minutes. Staurosporine (1.0 equivalent) was additionally added and stirred at room temperature for 16 hours. After confirming that the reaction was completed through LC-MS, the target compound was obtained by purification through prep-HPLC. (Yield: 33%) MS (ESI): m/z 965 [M+1] ⁺

Example 46. Preparation of compound 46

The compound of Example 46 according to the present invention was prepared in a similar manner to Example 45. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

Example 47. Preparation of compound 47

Step 1: Preparation of tert-butyl4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperazine-1-carboxylate

2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (1.0 equiv.) was dissolved in DMSO (0.3 M) in a reaction mixture of tert-butylpiperazine- 1-carboxylate (1.0 equiv.), DIPEA (3.0 equiv.) was added and stirred at 130° C. for 1 hour. After confirming the completion of the reaction through LC-MS, it was put in ice water, stirred for 30 minutes, and filtered to obtain the target compound. (Yield: 100%) MS (ESI): m/z 443 [M+1] ⁺

Step 2: Preparation of 2-(2,6-dioxopiperidin-3-yl)-4-(piperazin-1-yl)isoindoline-1,3-dione

tert-Butyl4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperazine-1-carboxylate prepared in step 1 ( 1.0 equivalent) was added to the reaction mixture in DCM (0.1 M), and TFA (10.0 equivalent) was added, followed by stirring at room temperature for 1 hour. After confirming that the reaction was completed through LC-MS, the target compound was obtained without further purification by concentration under reduced pressure. MS (ESI): m/z 343 [M+1] ⁺

Step 3: tert-Butyl3-(2-(2-(2-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl) ) piperazin-1-yl) ethoxy) ethoxy) ethoxy) propanoate

2-(2,6-dioxopiperidin-3-yl)-4-(piperazin-1-yl)isoindoline-1,3-dione (1.0 eq.) prepared in step 2 was mixed with DMF (0.1 M ), tert-butyl3-(2-(2-(2-iodoethoxy)ethoxy)ethoxy)propanoate (1.0 equiv.) and potassium carbonate (2.0 equiv.) were added to the reaction mixture dissolved in was stirred for 6 hours. After confirming that the reaction was completed through LC-MS, distilled water was added, the organic material was extracted with EA, the organic layer was washed with brine, the remaining water was removed with _{Na 2} SO _{4 , filtered, and concentrated.} The target compound was obtained without further purification. MS (ESI): m/z 603 [M+1] ⁺

Step 4: 3-(2-(2-(2-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperazine) Preparation of -1-yl) ethoxy) ethoxy) ethoxy) propanoic acid

tert-Butyl3-(2-(2-(2-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4 prepared in step 3) -yl)piperazin-1-yl)ethoxy)ethoxy)ethoxy)propanoate (0.1 eq) was dissolved in DCM (0.1 M), TFA (10 eq) was added to the reaction mixture and stirred at room temperature for 2 hours. did. After confirming that the reaction was complete through LC-MS, it was concentrated under reduced pressure and purified through prep-HPLC to obtain the target compound. (Yield: 21%) MS (ESI): m/z 547 [M+1] ⁺

Step 5: 3-(2-(2-(2-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperazine) -1-yl)ethoxy)ethoxy)ethoxy)-N-((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15 , 16-hexahydro-5H, 14H-17-oxa-4b, 9a, 15-triaza-5,9-methanodibenzo [b, h] cyclonona [jkl] cyclopenta [e] -as-indacene Preparation of -7-yl)-N-methylpropanamide

3-(2-(2-(2-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl) prepared in step 4) Piperazin-1-yl) ethoxy) ethoxy) ethoxy) propanoic acid (1.0 equiv.) was dissolved in DMF (0.1 M) to a reaction mixture, DIPEA (3.0 equiv.) and HATU (2.0 equiv.) were added to 1 at room temperature. time was stirred. Staurosporine (1.0 equivalent) was additionally added and stirred at room temperature for 16 hours. After confirming that the reaction was completed through LC-MS, the target compound was obtained by purification through prep-HPLC. (Yield: 65%) MS (ESI): m/z 996 [M+1] ⁺

Examples 48 to 49. Preparation of compounds 48 to 49

The compounds of Examples 48 to 49 according to the present invention were prepared in a manner similar to that of Example 47. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

Example 50. Preparation of compound 50

Step 1: tert-Butyl12-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperazin-1-yl)- Preparation of 12-oxododecanoate

After dissolving 2-(2,6-dioxopiperidin-3-yl)-4-(piperazin-1-yl)isoindoline-1,3-dione (1.0 equivalent) in DMF (0.1 M), 12-(tert-butoxy)-12-oxododecanoic acid (1.1 eq.), HATU (2.0 eq.), and DIPEA (4.0 eq.) were added and stirred at room temperature for 2 hours. Water and brine were added to the reaction mixture, and organics were extracted with EtOAc. The collected organic layers were concentrated after removing the remaining water using _{Na 2} SO _{4 .} The obtained target compound as a brown liquid was used in the next reaction without further purification. (Yield: 100%)

MS (ESI): m/z 555 [M-(t-Bu)+1] ⁺

Step 2: 12-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperazin-1-yl)-12-oxo Preparation of dodecanoic acid

tert-Butyl12-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperazin-1-yl prepared in step 1 )-12-oxododecanoate (1.0 eq.) was dissolved in DCM (0.4 M), TFA (30 eq.) was added and stirred at room temperature for 2 hours. The reaction mixture was concentrated and purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 41 %) MS (ESI): m/z 555 [M+1] ⁺

Step 3: 12-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperazin-1-yl)-N-( (5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a, Preparation of 15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)-N-methyl-12-oxododecanamide

12-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperazin-1-yl)-12 prepared in step 2 -Oxododecanoic acid (1.0 equiv.) was dissolved in DMF (0.05 M), Staurosporine (1.0 equiv.), HATU (2.0 equiv.), and DIPEA (4.0 equiv.) were added and stirred at room temperature for 2 hours. Water and brine were added to the reaction mixture, and organics were extracted with EtOAc. The collected organic layers were concentrated after removing the remaining water using _{Na 2} SO _{4 .} The reaction mixture was purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 61 %) MS (ESI): m/z 1004 [M+1] ⁺

Examples 51 to 52. Preparation of compounds 51 to 52

The compounds of Examples 51 to 52 according to the present invention were prepared in a manner similar to that of Example 50. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

Example 53. Preparation of compound 53

Example 53-1

Step 1: Preparation of tert-butyl(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl)carbamate

2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (1.0 equiv), tert-butyl(2-aminoethyl)carbamate (1.2 equiv) After dissolving in NMP (0.1 M), DIPEA (3.0 equivalents) was added and stirred at 90 °C for 12 hours. EtOAc and 10% citric acid were added to the reaction mixture, and organic matter was extracted. The collected organic layers were concentrated after removing the remaining water using _{Na 2} SO _{4 .} The reaction mixture was purified through MPLC to obtain the title compound as a yellow solid. (Yield: 33%) MS (ESI): m/z 417 [M-(t-Bu)+1] ⁺

Step 2: Preparation of 4-((2-aminoethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

tert-butyl(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl)carbamate ( 1.0 equiv) was dissolved in DCM (0.4 M), and TFA (30 equiv.) was added thereto, followed by stirring at room temperature for 30 minutes. After adding ether to the reaction mixture, it was filtered to obtain the target compound as a yellow solid. (Yield: 100%) MS (ESI): m/z 317 [M+1] ⁺

Example 53-2

Step 1: Methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9- Preparation of methanodibenzo [b, h] cyclonona [jkl] cyclopenta [e] -as-indasen-7-yl) (methyl) amino) dodecanoate

Staurosporine (1.0 equiv.) was dissolved in DMF (0.1 M), tert-butyl12-bromododecanoate (1.1 equiv.) and TEA (2.0 equiv.) were added, followed by stirring at 80 °C for 16 hours. Water and brine were added to the reaction mixture, and organics were extracted with EtOAc. The collected organic layers were concentrated after removing the remaining water using _{Na 2} SO _{4 .} The reaction mixture was purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 8.4%) MS (ESI): m/z 665 [M-(t-Bu)+1] ⁺

Step 2: 12-(((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17 -oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)(methyl)amino)dode manufacture of cansan

Methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5 prepared in step 1, 9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)(methyl)amino)dodecanoate (1.0 equiv.) was dissolved in hydrochloric acid (4.0M 1,4 -Dioxane solution, 1mL) and stirred at room temperature for 1 hour. The target compound as a brown liquid obtained after concentration was used in the next reaction without further purification. MS (ESI): m/z 665 [M+1] ⁺

Step 3: N-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl)-12-(((( 5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15 Preparation of -triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)(methyl)amino)dodecanamide

12-(((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H prepared in step 2 -17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)(methyl)amino ) After dissolving dodecanoic acid (1.0 equiv.) in DMF (0.05 M), HATU (3.0 equiv.), 4-((2-aminoethyl)amino)-2-(2,6-dioxopiperidin-3-yl ) Isoindoline-1,3-dione (1.2 equivalents) and DIPEA (2.0 equivalents) were added and stirred at room temperature for 1 hour. Water and brine were added to the reaction mixture, and organics were extracted with EtOAc. The collected organic layers were concentrated after removing the remaining water using _{Na 2} SO _{4 .} The reaction mixture was purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 33%) MS (ESI): m/z 964 [M+1] ⁺

Example 54. Preparation of compound 54

Step 1: tert-Butyl5-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl)amino) Preparation of 5-oxopentanoate

After dissolving 4-((2-aminoethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (1.0 equiv.) in DMF (0.1 M) , 5-(tert-butoxy)-5-oxopentanoic acid (1.1 eq.), HATU (2.0 eq.), and DIPEA (4.0 eq.) were added and stirred at room temperature for 2 hours. Water and brine were added to the reaction mixture, and organics were extracted with EtOAc. The collected organic layers were concentrated after removing the remaining water using _{Na 2} SO _{4 .} The obtained target compound as a brown liquid was used in the next reaction without further purification. (Yield: 100%)

MS (ESI): m/z 431 [M-(t-Bu)+1] ⁺

Step 2: 5-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl)amino)-5- Preparation of oxopentanoic acid

tert-Butyl5-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl) prepared in step 1 Amino)-5-oxopentanoate (1.0 equiv.) was added with hydrochloric acid (4M 1,4-dioxane solution, 20 equiv), followed by stirring at room temperature for 1 hour. The reaction mixture was concentrated and purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 34%) MS (ESI): m/z 431 [M+1] ⁺

Step 3: N ^One -(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl)-N ⁵ -((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b, 9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)-N ⁵ -Preparation of methylglutaramide

5-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl)amino)- prepared in step 2 After 5-oxopentanoic acid (1.0 equiv.) was dissolved in DMF (0.05 M), Staurosporine (1.0 equiv.), HATU (2.0 equiv.), and DIPEA (3.0 equiv) were added and stirred at room temperature for 2 hours. Water and brine were added to the reaction mixture, and organics were extracted with EtOAc. The collected organic layers were concentrated after removing the remaining water using _{Na 2} SO _{4 .} The reaction mixture was purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 43%) MS (ESI): m/z 879 [M+1] ⁺

Examples 55 to 57. Preparation of compounds 55 to 57

The compounds of Examples 55 to 57 according to the present invention were prepared in a manner similar to that of Example 54. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

Example 58. Preparation of compound 58

Step 1: Preparation of tert-butyl(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethyl)carbamate

2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (1.0 equivalent) was dissolved in THF (0.1 M) in a reaction mixture of tert-butyl (2- Hydroxyethyl)carbamate (1.0 equiv), PPh ₃ (1.5 equiv) was added. DIAD (2.0 equiv) was added dropwise at 0 °C and stirred at -15 °C for 16 h. After confirming that the reaction was completed through LC-MS, distilled water was added, the organic material was extracted three times with EA, the organic layer was washed with brine, the remaining water was removed with _{Na 2} SO _{4 , filtered, and concentrated.} The target compound was obtained without further purification. MS (ESI): m/z 318 [M+1] ⁺

Step 2: Preparation of 4-(2-aminoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

tert-butyl(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethyl)carbamate prepared in step 1 ( 1.0 equivalent) was added to the reaction mixture in DCM (0.1 M), and TFA (10.0 equivalent) was added, followed by stirring at room temperature for 1 hour. After confirming that the reaction was completed through LC-MS, the target compound was obtained by purification through prep-HPLC. (Yield: 38%) MS (ESI): m/z 318 [M+1] ⁺

Step 3: tert-Butyl5-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethyl)amino) Preparation of 5-oxopentanoate

4-(2-aminoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (1.0 equiv.) prepared in step 2, 5-(tert- DIPEA (3.0 equivalents) and HATU (2.0 equivalents) were added to a reaction mixture in which butoxy)-5-oxopentanoic acid (1.0 equivalent) was dissolved in ACN (0.1 M), followed by stirring at room temperature for 3 hours. After confirming that the reaction was completed through LC-MS, the target compound was obtained without further purification by concentration under reduced pressure. MS (ESI): m/z 488 [M+1] ⁺

Step 4: 5-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethyl)amino)-5- Preparation of oxopentanoic acid

tert-Butyl5-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethyl) prepared in step 3 To a reaction mixture of amino)-5-oxopentanoate (1.0 eq.) dissolved in DCM (0.1 M), TFA (10 eq.) was added, followed by stirring at room temperature for 2 hours. After confirming that the reaction was complete through LC-MS, it was concentrated under reduced pressure and purified through prep-HPLC to obtain the target compound. (Yield: 58%) MS (ESI): m/z 432 [M+1] ⁺

Step 5: N ^One -(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethyl)-N ⁵ -((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b, 9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[Jkl]cyclopenta[e]-as-indasen-7-yl)-N ⁵ -Preparation of methylglutaramide

5-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethyl)amino)- prepared in step 4 DIPEA (3.0 equiv.) and HATU (2.0 equiv.) were added to a reaction mixture in which 5-oxopentanoic acid (1.0 equiv.) was dissolved in DMF (0.1 M), followed by stirring at room temperature for 1 hour. Staurosporine (1.0 equivalent) was additionally added and stirred at room temperature for 3 hours. After confirming that the reaction was completed through LC-MS, the target compound was obtained by purification through prep-HPLC. (Yield: 78%) MS (ESI): m/z 880 [M+1] ⁺

Examples 59 to 61. Preparation of compounds 59 to 61

The compounds of Examples 59 to 61 according to the present invention were prepared in a manner similar to that of Example 58. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

Example 62. Preparation of compound 62

Step 1: Preparation of 4-(2-chloroethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (1.0 equiv), 1-bromo-2-chloroethane (1.4 equiv), hydrogen carbonate Sodium (6.0 equivalents) and potassium iodide (0.6 equivalents) were dissolved in DMF and stirred at 60 ^o C for 2 hours. After confirming the product by LC-MS, it was purified by prep-HPLC to obtain the target compound. (Yield: 27%)

Step 2: Preparation of 4-(2-azidoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

4-(2-chloroethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (1.0 equiv.) prepared in step 1, sodium azide ( 10 equivalents) was dissolved in anhydrous DMF and stirred at 90 ^° C for 16 hours. After confirming the product by LC-MS, it was filtered through Celite, and the filtrate was diluted with water and extracted with EA. After removing the remaining water with Na ₂ SO ₄ , filtration, and concentration, the next reaction proceeded without further purification.

Step 3: 5-(1-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethyl)-1H- Preparation of 1,2,3-triazol-4-yl)pentanoic acid

4-(2-azidoethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (1.0 equiv) prepared in step 2, hepta-6- Inosic acid (1.1 eq) was dissolved in DCM:t-BuOH (1:2). Sodium ascorbate (2.2 equiv) and copper(II) sulfate hydrate (0.3 equiv) were dissolved in water and then added to the mixture. The reaction mixture ^{was stirred at 60 °} C for 4 h. After confirming the product by LC-MS, it was diluted in water and extracted with DCM. The _{remaining water was removed with Na 2} SO ₄ , filtered, concentrated, and purified by MPLC to obtain the target compound. (Two-step yield: 43%) MS (ESI): m/z 470 [M+1] ⁺

Step 4: 5-(1-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethyl)-1H- 1,2,3-triazol-4-yl)-N-((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15, 16-hexahydro-5H, 14H-17-oxa-4b, 9a, 15-triaza-5,9-methanodibenzo [b, h] cyclonona [jkl] cyclopenta [e] -as-indacene- Preparation of 7-yl)-N-methylpentanamide

5-(1-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)ethyl)- prepared in step 3 Dissolve 1H-1,2,3-triazol-4-yl)pentanoic acid (1.0 equiv.) and HATU (3.0 equiv.) in DMF, and simultaneously dissolve Staurosporine (1.0 equiv.) and DIPEA (2.0 equiv.) in DMF for 30 minutes each. stirred. The two reaction mixtures were mixed and stirred at room temperature for 1 hour. The product was confirmed by LC-MS and purified by prep-HPLC to obtain the target compound. (Yield: 45%) MS (ESI): m/z 918 [M+1] ⁺

Example 63. Preparation of compound 63

The compound of Example 63 according to the present invention was prepared in a manner similar to that of Example 62. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

Example 64. Preparation of compound 64

Step 1: 3-methoxy-4-(((5R,7R,8R,9S)-8-methoxy-9-methyl-16-oxo-6,7,8,9,15,16-hexahydro- 5H,14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl) ( Preparation of methyl)amino)cyclobut-3-ene-1,2-dione

Staurosporine (1.0 equiv.) was dissolved in DCM:MeOH (2:1), followed by addition of 3,4-dimethoxycyclobut-3-ene-1,2-dione (2.0 equiv.) and TEA (3.0 equiv.) The mixture was stirred at ℃ for 3 hours. The reaction mixture was concentrated and purified by prep-HPLC to obtain the target compound as a white solid. (Yield: 49%)

MS (ESI): m/z 577 [M+2] ²⁺

Step 2: tert-Butyl(2-(2-(2-((2-(((5R,7R,8R,9S)-8-methoxy-9-methyl-16-oxo-6,7,8; 9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as Preparation of -indacen-7-yl) (methyl) amino) -3,4-dioxocyclobut-1-en-1-yl) amino) ethoxy) ethoxy) ethyl) carbamate

3-methoxy-4-(((5R,7R,8R,9S)-8-methoxy-9-methyl-16-oxo-6,7,8,9,15,16-hexa prepared in step 1 Hydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indacen-7-yl ) (methyl) amino) cyclobut-3-ene-1,2-dione (1.0 eq.) was dissolved in EtOH, followed by tert-butyl (2- (2- (2-aminoethoxy) ethoxy) ethyl) carba Mate (1.5 eq.) and TEA (3.0 eq.) were added and stirred at 50° C. for 3 hours. After the reaction mixture was concentrated, the target compound obtained as a yellow liquid was used in the next reaction without further purification. (Yield: 100%) MS (ESI): m/z 793 [M+1] ⁺

Step 3: 3-((2-(2-(2-aminoethoxy)ethoxy)ethyl)amino)-4-(((5R,7R,8R,9S)-8-methoxy-9-methyl- 16-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona Preparation of [jkl] cyclopenta [e] -as-indacen-7-yl) (methyl) amino) cyclobut-3-ene-1,2-dione

tert-butyl (2-(2-(2-((2-(((5R,7R,8R,9S)-8-methoxy-9-methyl-16-oxo-6,7, 8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e] -as-indacen-7-yl)(methyl)amino)-3,4-dioxocyclobut-1-en-1-yl)amino)ethoxy)ethoxy)ethyl)carbamate (1.0 equiv) After dissolving in DCM (0.1 M), TFA (30 equivalents) was added and stirred at room temperature for 1 hour. After adding ether to the reaction mixture, it was filtered to obtain the target compound as a yellow solid. (Yield: 100%) MS (ESI): m/z 693 [M+1] ⁺

Step 4: 2-(2,6-dioxopiperidin-3-yl)-4-((2-(2-(2-((2-(((5R,7R,8R,9S)-8-) Methoxy-9-methyl-16-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo [b,h]cyclonona[jkl]cyclopenta[e]-as-indacen-7-yl)(methyl)amino)-3,4-dioxocyclobut-1-en-1-yl)amino) Preparation of ethoxy) ethoxy) ethyl) amino) isoindoline-1,3-dione

3-((2-(2-(2-aminoethoxy)ethoxy)ethyl)amino)-4-(((5R,7R,8R,9S)-8-methoxy-9- prepared in step 3 Methyl-16-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h] After dissolving cyclonona [jkl] cyclopenta [e] -as-andacen-7-yl) (methyl) amino) cyclobut-3-ene-1,2-dione (1.0 equiv.) in DMSO (0.1 M) , K ₂ CO ₃ (3.0 equiv.), 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (1.0 equiv.) was added to 4 at 80 °C. stirred for hours. Water and brine were added to the reaction mixture, and organics were extracted with EtOAc. The collected organic layers were concentrated after removing the remaining water using _{Na 2} SO _{4 .} The reaction mixture was purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 3%)

MS (ESI): m/z 949 [M+1] ⁺

Example 65. Preparation of compound 65

Step 1: 2-(2,6-dioxopiperidin-3-yl)-4-((2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl)amino) Preparation of isoindoline-1,3-dione

2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (1.0 equiv.), 2-(2-(2-(2) in DMF (0.05 M) -Aminoethoxy)ethoxy)ethoxy)ethan-1-ol (1.5 eq.) and DIPEA (3.0 eq.) were dissolved, and the mixture was stirred at 80° C. for 4 hours. LC-MS confirmed the desired product. After cooling to room temperature, the mixture was purified using prep-HPLC to obtain the target compound as a yellow solid. (Yield: 50%)

MS (ESI): m/z 450 [M+1] ⁺

Step 2: 2-(2,6-dioxopiperidin-3-yl)-4-((2-(2-(2-(2-iodoethoxy)ethoxy)ethoxy)ethyl)amino) Preparation of isoindoline-1,3-dione

2-(2,6-dioxopiperidin-3-yl)-4-((2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl) prepared in step 1 After dissolving amino)isoindoline-1,3-dione (1.0 eq.) in dichloromethane (0.2 M), triphenylphosphine followed by imidazole was added to the mixture. Iodine was added little by little to the mixture and stirred at room temperature overnight. LC-MS identified the desired product, the mixture _{was diluted with H 2} O and extracted with DCM. The combined organic layers were dried over MgSO ₄ , filtered, and the filtrate was concentrated to proceed with the next reaction without further purification. MS (ESI): m/z 560 [M+1] ⁺

Step 3: tert-Butyl1-(2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-) Preparation of yl) amino) ethoxy) ethoxy) ethoxy) ethyl) piperidine-4-carboxylate

2-(2,6-dioxopiperidin-3-yl)-4-((2-(2-(2-(2-iodoethoxy)ethoxy) prepared in step 2 in DMF (0.05 M) ) ethoxy) ethyl) amino) isoindoline-1,3-dione (1.0 equiv.), tert-butylpiperidine-4-carboxylate (1.2 equiv.), K ₂ CO ₃ (2.0 equiv.) , the mixture was stirred at 80 °C for 1 hour. LC-MS confirmed the desired product. After cooling to room temperature, the mixture was subjected to prep-HPLC to obtain the target compound as a yellow solid. (Yield: 24%) MS (ESI): m/z 617 [M+1] ⁺

Step 4: 1-(2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino) Preparation of ) ethoxy) ethoxy) ethoxy) ethyl) piperidine-4-carboxylic acid

tert-Butyl1-(2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-) prepared in step 3 4M HCl (10 equivalents) dissolved in dioxane was added to 4-yl)amino)ethoxy)ethoxy)ethoxy)ethyl)piperidine-4-carboxylate, and the mixture was stirred at room temperature for 1 hour. The desired product was identified through LC-MS, and the mixture was concentrated and used in the next reaction without further purification. MS (ESI): m/z 561 [M+1] ⁺

Step 5: 1-(2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino) )ethoxy)ethoxy)ethoxy)ethyl)-N-((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16 - Hexahydro-5H, 14H-17-oxa-4b, 9a, 15-triaza-5,9-methanodibenzo [b, h] cyclonona [jkl] cyclopenta [e] -as-indacene-7 Preparation of -yl)-N-methylpiperidine-4-carboxamide

1-(2-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl) prepared in step 4) )Amino)ethoxy)ethoxy)ethoxy)ethyl)piperidine-4-carboxylic acid (1.0 equiv.) in DMF (0.2 M) was added with HATU (1.5 equiv.), followed by 30 °C at 50 °C. stirred for minutes. After adding staurosporine (1.0 eq.) and DIPEA (3.0 eq.) to the mixture, the mixture was stirred at 50 °C for 1 hour. LC-MS confirmed the desired product. The mixture was subjected to prep-HPLC to obtain the target compound as a yellow solid. (Yield: 23%) MS (ESI): m/z 1009 [M+1] ⁺

Example 66. Preparation of compound 66

Step 1: Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione

5-fluoroisobenzofuran-1,3-dione (1.0 equiv.), 3-aminopiperidine-2,6-dione hydrochloride (1.1 equiv.) and potassium acetate (3.1 equiv) were dissolved in AcOH (0.05 M). Then, the mixture was stirred at 90 °C for 4 hours. LC-MS confirmed the desired product. Water was added to the mixture at 0° C., and the obtained solid was filtered and dried under vacuum to give the title compound as a gray solid. (Yield: 96%) MS (ESI): m/z 277 [M+1] ⁺

Step 2: Preparation of tert-butyl1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carboxylate

2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (1.0 equiv) prepared in step 1 in DMF (0.05 M), tert-butylpiperidin After dissolving din-4-carboxylate (1.5 equivalents) and DIPEA (3.0 equivalents), the mixture was stirred at 80° C. for 2 hours. LC-MS confirmed the desired product. The mixture _{was diluted with H 2} O and extracted with EA. The combined organic layers were dried over MgSO ₄ , filtered, and the filtrate was concentrated and purified by MPLC. The target compound as a yellow solid was obtained. (Yield: 96%)

MS (ESI): m/z 442 [M+1] ⁺

Step 3: Preparation of 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carboxylic acid

tert-Butyl1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carboxylate prepared in step 2 After adding 4M HCl (10 eq) dissolved in dioxane, the mixture was stirred at room temperature for 1 hour. The desired product was identified through LC-MS, and the mixture was concentrated and used in the next reaction without further purification. MS (ESI): m/z 386 [M+1] ⁺

Step 4: 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-N-((5S,6R,7R,9R)- 6-Methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9-meta Preparation of nodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)-N-methylpiperidine-4-carboxamide

1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carboxylic acid prepared in step 3 (1.0 equiv. ) was added to a solution of HATU (1.5 equivalents) in DMF (0.2 M), followed by stirring at 50 °C for 30 minutes. Staurosporine (1.0 equiv.) and DIPEA (3.0 equiv) were added to the mixture, followed by stirring at 50° C. for 1 hour. LC-MS confirmed the desired product. The mixture was subjected to prep-HPLC to obtain the target compound as a yellow solid. (Yield: 23%)

MS (ESI): m/z 834 [M+1] ⁺

Example 67. Preparation of compound 67

Step 1: tert-Butyl(3-(((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H , 14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)(methyl Preparation of )carbamoyl)bicyclo[1.1.1]pentan-1-yl)carbamate

3-(tert-butoxycarbonyl)bicyclo[1.1.1]pentane-1-carboxylic acid (1.0 equiv.) and Staurosporine (1.1 equiv.) were dissolved in ACN 0.1M, followed by HATU (2.5 equiv.), DIPEA ( 3.0 equivalent) and stirred at room temperature for 2 hours. After confirming that the reaction was complete by LC-MS, the solvent was concentrated under reduced pressure. The concentrated compound was dissolved in EA (0.01M), filtered through a Celite filter, and concentrated again under reduced pressure. The obtained material was purified by MPLC to obtain the target compound. (Yield: 89%) MS (ESI): m/z 676 [M+1] ⁺

Step 2: 3-Amino-N-((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H, 14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)-N- Preparation of methylbicyclo[1.1.1]pentane-1-carboxamide

The compound prepared in step 1 was dissolved in 4M HCl/1,4-dioxane (0.05M) and stirred at room temperature for 4 hours. After confirming that the reaction was complete by LC-MS, it was concentrated under reduced pressure and used in the next reaction without further purification. MS (ESI): m/z 576 [M+1] ⁺

Step 3: 3-(3-(2-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino) )ethoxy)ethoxy)ethoxy)propanamido)-N-((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15 , 16-hexahydro-5H, 14H-17-oxa-4b, 9a, 15-triaza-5,9-methanodibenzo [b, h] cyclonona [jkl] cyclopenta [e] -as-indacene Preparation of -7-yl)-N-methylbicyclo[1.1.1]pentane-1-carboxamide

Compound 3-amino-N-((5S,6R,7R,9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro prepared in step 2 -5H,14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl) -N-Methylbicyclo[1.1.1]pentane-1-carboxamide (1.0 eq.) and 3-(2-(2-(2-((2-(2,6-dioxopiperidine-3-) yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)propanoic acid (1.0 equiv.) was dissolved in DMF (0.2M), followed by HATU (2.5 equiv.), DIPEA ( 6.0 equivalents) was added and stirred at room temperature for 2 hours. After confirming that the reaction was completed by LC-MS, the target compound was obtained by using prep-HPLC. (Yield: 22%) MS (ESI): m/z 1036 [M+1] ⁺

Example 68. Preparation of compound 68

The compound of Example 68 according to the present invention was prepared in a manner similar to that of Example 67. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

Example 69. Preparation of compound 69

Step 1: tert-Butyl 1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-3,6,9,12 -Preparation of tetraoxapentadecane-15-oate

2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (1.0 equiv), tert-butyl1-iodo-3,6,9,12- Tetraoxapentadecane-15-oate (1.1 eq.), Cs ₂ CO ₃ (2.5 eq.) was dissolved in DMF (0.1 M) and stirred at room temperature for 2 hours. After stirring at 50° C. for an additional 2 hours, water and brine were added, and the organic material was extracted with EtOAc. The collected organic layer was concentrated after removing the remaining water using _{Na 2} SO _{4 .} The reaction mixture was purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 22%) MS (ESI): m/z 523 [M-(t-Bu)+1] ⁺

Step 2: 1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-3,6,9,12-tetraoxa Preparation of pentadecane-15-oic acid

tert-Butyl1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-3,6,9 prepared in step 1 ,12-tetraoxapentadecane-15-oate (1.0 equivalent) was dissolved in 4 N HCl/1,4-Dioxane (0.1 M) and stirred at room temperature for 1 hour. The reaction mixture was concentrated and used in the next reaction without further purification.

MS (ESI): m/z 523 [M+1] ⁺

Step 3: 1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-N-((5S,6R,7R, 9R)-6-methoxy-5-methyl-14-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5, 9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)-N-methyl-3,6,9,12-tetraoxapentadecane-15- Preparation of amides

1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-3,6,9,12- prepared in step 2 Tetraoxapentadecane-15-oic acid (1.0 eq.), Staurosporine (1.1 eq.), HATU (2.0 eq.), and DIPEA (4.0 eq.) were dissolved in DMF (0.01 M) and stirred at room temperature for 1 hour. Water and brine were added, and organic matter was extracted with EtOAc. The collected organic layer was concentrated after removing the remaining water using _{Na 2} SO _{4 .} The reaction mixture was purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 17%) MS (ESI): m/z 971 [M+1] ⁺

Example 70. Preparation of compound 70

The compound of Example 70 according to the present invention was prepared in a manner similar to that of Example 69. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

Example 71. Preparation of compound 71

The compound of Example 71 according to the present invention was prepared in a similar manner to Example 45. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

Example 72. Preparation of compound 72

Step 1: tert-Butyl16-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazole- Preparation of 5-yl) -4,7,10,13-tetraoxahexadecanoate

tert-Butyl16-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl )-4,7,10,13-tetraoxahexadec-15-inoate (1.0 equivalent) was dissolved in ethyl acetate (0.1 M), 10% Pd/C was added, and the mixture was stirred at room temperature and under hydrogen for 4 hours. . LC-MS was used to identify the product, and the reaction mixture was filtered through a Celite filter. The filtered organic layer was concentrated under reduced pressure, and the target compound was obtained through MPLC. (Yield: 80%) MS (ESI): m/z 522 [M-(t-Bu)+1] ⁺

Step 2: 16-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl ) Preparation of -4,7,10,13-tetraoxahexadecanoic acid

tert-Butyl16-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imi prepared in step 1 Dazol-5-yl)-4,7,10,13-tetraoxahexadecanoate (1.0 equivalent) was dissolved in 4 N HCl/1,4-Dioxane (0.1 M) and stirred at room temperature for 1 hour. The reaction mixture was concentrated and used in the next reaction without further purification.

Step 3: 16-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl )-N-((5R,7R,8R,9S)-8-methoxy-9-methyl-16-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa -4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indasen-7-yl)-N-methyl4,7, Preparation of 10,13-tetraoxahexadecanamide

16-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazole-5 prepared in step 2 -yl)-4,7,10,13-tetraoxahexadecanoic acid (1.0 equiv.), Staurosporine (1.1 equiv.), HATU (2.0 equiv.), and DIPEA (4.0 equiv.) were dissolved in DMF (0.01 M) and then at room temperature Stirred for 1 hour. Water and brine were added, and organic matter was extracted with EtOAc. The collected organic layer was concentrated after removing the remaining water using _{Na 2} SO _{4 .} The reaction mixture was purified through prep-HPLC to obtain the target compound as a yellow solid. (Yield: 41%) MS (ESI): m/z 971 [M+1] ⁺

Example 73. Preparation of compound 73

The compound of Example 73 according to the present invention was prepared in a manner similar to that of Example 69. Chemical structural formulas of each Example are summarized in Table 1, and compound names, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

reference material. Preparation of reference substances

A reference material was prepared in a manner similar to that of Example 1. Chemical structural formulas of each Example compound and reference material are summarized in Table 1, and the compound name, ¹ H NMR, MS, HPLC data and yield are summarized in Table 3.

[Table 3]

Experimental Example 1. Evaluation of the enzyme inhibitory activity of the compound according to the present invention

In order to evaluate the inhibitory activity of the compound according to the present invention on the kinase, the following method was performed. Specifically, the substrate was prepared in a basic reaction buffer (20 mM Hepes (pH 7.5), 10 mM MgCl ₂ , 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM Na ₃ VO ₄ , 2 mM DTT, 1% DMSO) and then the cofactor required for the reaction was added. Next, after the corresponding kinase was added and mixed, each of the compounds prepared in Examples was added using Acoustic technology (Echo550; nanoliter range). After standing at room temperature for 20 minutes, 33P-ATP (specific activity 10 mCi/mL) was added to initiate the reaction. After reacting at room temperature for 2 hours, spotting was made on P81 exchange paper. After the reaction, kinase activity was detected using the filter-binding method.

Table 4 below summarizes the inhibitory activities of the compounds of Examples for AURKA, FLT3 (ITD) and c-KIT (D816V). _{When the IC 50} value of the measured kinase was less than 50 nM, it was classified as Grade A, when it was 50 or more and 100 nM or less, it was classified as Grade B, and when it exceeded 100 nM, it was classified as Grade C.

[Table 4]

Experimental Example 2. Evaluation of various kinase inhibitory activities of the compounds according to the present invention

In order to evaluate the inhibitory activity of the compounds according to the present invention to various enzymes, the following experiments were performed. Specifically, among the example compounds of this alias, the selected Example compound 1 was commissioned by DiscoverX to measure enzyme (kinase) selectivity, and the experiment was conducted using a ^{scanMAX TM Kinase analysis panel.}

At this time, the concentration of the drug treated with the enzyme was 1 uM in DMSO, and the control percentage (% control) was determined in the same way as in Equation 1 below.

[Equation 1]

(Example compound - positive control) / (negative control - positive control) × 100

Here, the positive control refers to a compound showing a percentage control of 0%, and the negative control indicates a percentage control of 100% with DMSO. In addition, the enzyme selectivity of the present invention was determined to have activity for each enzyme if the control percentage was <10% (ie, less than 10%) for each enzyme.

The compounds according to the present invention are ARK5, AURKA, AURKB, AURKC, CAMK1G, CAMK2A, CAMK2D, CAMKK1, CAMKK2, DYRK1A, EGFR(L858R,T790M), EGFR(T790M), FLT3(D835H), FLT3(D835V), FLT3(D835) D835Y), FLT3(ITD), FLT3(ITD,D835V), FLT3(ITD,F691L), FLT3(K663Q), FLT3(N841I), FLT3(R834Q), GRK1, GRK7, IKK-epsilon, IRAK3, JAK2(JH1domain) -catalytic), JAK3 (JH1domain-catalytic), KIT (A829P), KIT (D816H), KIT (D816V), MAP4K2, MARK1, MLK1, MLK3, MST2, PDGFRB, PHKG1, PKN1, PKN2, PLK4, PRKCE, PRKG2, PRKCE PYK2, QSK, RET (V804M), RSK2 (Kin.Dom.1-N-terminal), RSK4 (Kin.Dom.1-N-terminal), SIK2, SNARK, TBK1, TNK1, TRKA, TRKB, or YSK4 kinase It can be seen that the control percentage has a value smaller than 10% for . This indicates that the compound according to the present invention has inhibitory activity against the enzymes listed above, suggesting that there is a useful effect when used in diseases related to the enzymes listed above.

Experimental Example 3. Evaluation of cell inhibitory activity of the compound according to the present invention

MV4-11 cells were obtained from ATCC and cultured in Iscove's Modified Dulbecco's Media (IMDM) supplemented with 10% FBS, 100 U/mL penicillin and 100 μg/mL streptomycin at 37° C., 5% CO _{2 .} Molt-4 and Molm-14 cells were obtained from ATCC and Roswell Park Memorial Institute 1640 Media (RPMI-) supplemented with 10% FBS, 100 U/mL penicillin, and 100 μg/mL streptomycin _{at 37° C., 5% CO 2} 1640). HMC-1.2 cells were obtained from Sigma-Aldrich, Iscove's Modified Dulbecco's supplemented with 10% FBS, 100 U/mL penicillin and 100 μg/mL streptomycin, 1.2 mM α-thioglycerol _{at 37° C., 5% CO 2} Cultured in Media (IMDM). Aurora A (#14475), c-KIT antibody (#3308S) and GAPDH (#5174) antibodies were obtained from Cell Signaling Technologies. The FLT3 (#ab245116) antibody was obtained from Abcam.

Using MV4-11, Molt-4, and Molm-14 cell lines, 5,000 cells were cultured in 100 μl culture medium per well of a 96-well plate. For each compound, it was treated by sequentially diluting it at a ratio of 1/10 from the highest concentration of 1,000 mM, and then _{incubated at 37° C. CO 2 in an} incubator for 72 hours. The compound-treated plate was treated according to the standard procedure of Cell Titer Glo (Promega), band intensity was measured through a plate reader (BioTek Synergy2), and nonlinear regression (4 parameter) was performed using Prism software (GraphPad). _{GI 50} values were obtained through fitting.

Table 5 below shows the inhibitory activity of the example compounds against Molm-14 and MV4-11 cell lines. _{When the GI 50} value of the measured kinase was less than 5 nM, it was classified as A, 5 or more and less than 10 nM, B, 10 or more, 50 nM or less, C, and 50 nM or more, D.

[Table 5]

Table 6 below shows the inhibitory activity of the Example compounds against Molt-4 cell line. _{When the GI 50} value of the measured kinase was less than 100 nM, it was classified as A grade, when it was 100 or more and less than 500 nM, it was classified as B, 500 or more and less than 1000 nM was classified as C, and when it exceeded 1000 nM, it was classified as D grade.

[Table 6]

Experimental Example 4. Cell inhibitory activity of Ba / F3 - FLT3 (ITD), FLT3 (ITD / D835Y), FLT3 (ITD / F691L / D835Y), and EGFR (Del19 / T790M / C797S) cell lines of a compound according to the present invention evaluation

Ba/F3 cell lines expressing FLT3 mutant and EGFR (Del19/T790M/C797S) mutant genes were cultured in a culture medium containing 90% RPMI, 10% FBS, and antibiotics (Welgene). After putting 5,000 Ba/F3 cell lines into a TC-treated 96-well plate (SPL), 100 uL each, and then injecting 1 uL each of the test compound diluted in dimethyl sulfoxide (diluted by 3 points from 1 mM concentration, a total of 10 concentrations) . Thereafter, it was incubated for 72 hours in a cell culture incubator. After adding 50 uL of CellTiter Glo (Promega) solution and storing it at room temperature for 10 minutes, the luminescence intensity was measured using a reader (Envision, PerkinElmer). The luminescence intensity for each final concentration of the test compound was graphed, and the GI ₅₀ value was obtained using Prism5.0 (GraphPad) software. The parental Ba/F3 cell line without FLT3 mutant and EGFR (del19/T790M/C797S) mutant gene is mouseIL-3 ( R&D Systems) was cultured using a culture solution. The compound-treated plates were processed according to the standard procedure of CellTiter Glo (Promega), band intensity was measured through a plate reader (BioTek Synergy2), and nonlinear regression (4 parameter) was fitted using Prism software (GraphPad). The GI ₅₀ value was obtained through _{When the GI 50} value of the kinase measured at each Ba/F3 is less than 100 nM, it is classified as A, 100 or more and less than 500 nM, B, 500 or more and 1000 nM or less, C, and if it exceeds 1000 nM, it is classified as D, and the results are described below. Table 7 shows.

[Table 7]

Experimental Example 5. AURKA, FLT3 (ITD) or c-KIT (D816V) degradation evaluation in each cell line of Example compound 5 according to the present invention

MV4-11, Molt-4, HMC1.2, Molm14 cells (1 x 10 ⁶ ) were treated with the indicated compounds (Example compound 5) solubilized in DMSO for 4 hours. Cells were collected at 1500 rpm for 3 minutes. Cells were lysed with Roche propatase inhibitor complete cocktail and phosphatase inhibitor (10 mM sodium fluoride, 10 mM sodium pyrophosphate, 1 mM sodium orthovanadate and 20 mM β-glycerophosphate) in lysis buffer (25 mM Tris, 1% Triton, 0.25% deoxycholic acid). Total protein concentration was determined by Pierce BCA Protein Assay, and 10-20 μg of protein was loaded onto a 10% Tris-Glycine gel. After standard gel electrophoresis, the separated proteins were transferred to nitrocellulose by dry transfer. Immunoblots were then processed according to standard procedures and incubated with each antibody. Band intensities were quantified by Li-Cor's Image Studio software. The results are shown in FIGS. 1 to 6 .

As the present invention has been described in detail above, for those of ordinary skill in the art, it will be apparent that these specific descriptions are only preferred embodiments, and the scope of the present invention is not limited thereby. Accordingly, it is intended that the substantial scope of the present invention be defined by the appended claims and their equivalents.

Claims (11)

A protein kinase degradation inducing compound represented by the following formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula 1]

In Formula 1,
TL (Target Ligand) is

ego,
ELL (E3 Ligase Ligand) is an E3 ubiquitin ligase ligand selected from the group consisting of the following formulas A-1, B-1 and C-1 (wherein

indicates that it is covalently linked with the Linker),
[Formula A-1]

[Formula B-1]

{In the above formulas A-1 and B-1,

Is

,

,

,

,

and

a ring selected from the group consisting of;
X ₁ is a single bond, -C≡C-, -CH ₂ -, -NH-, -O-, -CO-, -COO-, -NHCO- or -CONH-;
X ₂ is -CH ₂ -, -CH(C _1-4 alkyl)-, -NH-, -N(C _1-4 alkyl)-, -O-, -CO-, -CH ₂ -CH ₂ -, -NH-CH ₂ -, -NH-CH(C _1-4 alkyl)-, -N=CH-, or -N=C(C _1-4 alkyl)-;
X ₃ is hydrogen or C _1-4 alkyl;
X ₄ is hydrogen, halogen, C _1-6 alkyl, CN, NH ₂ , NO ₂ , OH, COH, COOH or CF ₃ ,
X ₅ is -CO- or -CH ₂ -}
[Formula C-1]

{In the formula C-1,
n is an integer from 1 to 3;

is C _5-6 cycloalkyl, phenyl, 5-6 membered heterocycloalkyl or heteroaryl, wherein said heterocycloalkyl or heteroaryl contains 1 to 3 N, O or S atoms;
Y ₁ is hydrogen or C _1-4 alkyl;
Y ₂ is C _1-4 alkyl, hydroxy(C _1-4 alkyl), —(C _0-2 alkyl)-COH, C _3-8 cycloalkyl, or phenyl;
Y ₃ is hydrogen,

or

and {here,

represents a position at which Y ₃ is covalently linked to Formula C-1;
Y ₄ is hydrogen, halogen, C _1-4 alkyl, —O(C _1-4 alkyl), C _3-6 cycloalkyl or 4-6 membered heterocycloalkyl [wherein one hydrogen of _{Y 4 is halogen,} -OH, -CN, -NHCOH, -NHCOCH ₃ , -COH or -COCH ₃ may be substituted];
Y ₅ is hydrogen or C _1-4 alkyl;

is 5 to 12 membered heterocycloalkyl or heteroaryl, wherein said heterocycloalkyl contains at least one N atom;
Y ₆ is C _1-6 alkyl, halogen or =O};
Linker is a device that connects TL and ELL. The method of claim 1,
ELL represented by Formula A-1 is a CRBN E3 ubiquitin ligase ligand represented by Formula A-2, a protein kinase degradation inducing compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula A-2]

In Formula A-2,
X ₂ is —CH ₂ —, —CH(C _1-4 alkyl)- or —CO—;
X ₃ is hydrogen or C _1-4 alkyl. The method of claim 1,
ELL represented by Formula B-1 is a CRBN E3 ubiquitin ligase ligand represented by Formula B-2, a protein kinase degradation inducing compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula B-2]

In Formula B-2,
X ₂ is -NH- or -N(C _1-4 alkyl)-;
X ₃ is hydrogen or C _1-4 alkyl. The method of claim 1,
ELL represented by Formula C-1 is a VHL E3 ubiquitin ligase ligand selected from the group consisting of Formulas C-2-1 and C-2-2, a protein kinase degradation inducing compound, a stereoisomer thereof, or a pharmaceutical thereof Naturally acceptable salts:
[Formula C-2-1]

[Formula C-2-2]

In the above formulas C-2-1 and C-2-2,

is a 5-membered member selected from the group consisting of oxazole, isoxazole, thiazole, isothiazole, imidazole, pyrazole, triazole, oxadiazole, pyrrole, pyrrolidine, furan, dihydrofuran and tetrahydrofuran a heteroaryl ring;
Y ₁ is hydrogen or C _1-3 alkyl;
Y _{4 is hydrogen or C 1-4} alkyl or C _3-5 cycloalkyl which may be substituted with halogen. The method of claim 1,
Linker is a protein kinase degradation inducing compound represented by the following formula L-1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula L-1]

In the above formula,
L _i is each independently a single bond, -CH ₂ -, -NH-, -O-, -S-, -SO-, -SO ₂ -, -CO-, -CH ₂ CH ₂ -, -CHCH-, -C≡C-, -CH ₂ CH ₂ O-, _{-OCH 2} CH ₂ -, -CH ₂ CH ₂ S-, -SCH ₂ CH ₂ -, -COO-, -CONH-, -NHCO- and

is a group selected from the group consisting of {wherein hydrogen in the group may be substituted with halogen, hydroxy, CF ₃ , C _1-6 alkyl or C _3-6 cycloalkyl};

is 3-14 membered cycloalkyl, heterocycloalkyl, arene or heteroarene;
p is an integer from 0 to 30; 6. The method of claim 5,
Linker is a protein kinase degradation inducing compound represented by the following formula L-2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula L-2]

In the above formula,
L _T is connected to it

Via a TL moiety covalently bonded defined in formula (I) and, L _E is associated with it

Covalently binds to the ELL moiety defined in Formula 1 through
L _T and L _E are each independently -L _X1 - or -L _X1 -L _C -L _X2 -,
L _X1 and L _X2 are each independently a single bond, -CH ₂ -, -O-, -NH-, -CO-, -NHCO- or -CONH-,
L _C is C _1-10 alkyl, C _6-14 arene, 5-14 membered heteroarene, C _3-12 cycloalkyl or 5-14 membered heterocycloalkyl {wherein said alkyl, arene, heteroarene, cycloalkyl or heterocycloalkyl may be substituted with halogen, C _1-6 alkyl, C _1-6 haloalkyl, C _1-5 alkoxy, C _1-6 aminoalkyl, =O or =S},
each L _ii is independently -(CH ₂ )-, -CHCH-, -C≡C-, -CH ₂ CH ₂ O- or _{-OCH 2} CH ₂ -, wherein at least one hydrogen in _{L ii is C 1 -6} alkyl, halo, C _1-6 haloalkyl, C _1-6 aminoalkyl},
s is an integer from 0 to 20;
q and r are each independently an integer from 0 to 10. 7. The method of claim 6,
In Formula L-2, L _T is a single bond, -CO(CH ₂ ) _a -,

,

,

,

or

and {wherein L _T , TL represents the TL moiety defined in Formula 1;

indicates that it is covalently linked to the remaining moieties of formula L-2},
L _E is a single bond, -CH ₂ -, -NH-, -O-,

,

, -L _Y -(CH ₂ ) _a -NH-, -L _Y -CH ₂ CH ₂ O-, -L _Y -CH ₂ CH ₂ NH- or

and {wherein L _E above, ELL represents the ELL moiety defined in Formula 1,

indicates that it is covalently linked to the remaining moieties of formula L-2},
L _Y are each independently a single bond, -(CH ₂ ) _a -, -CO-, -O-, -NH- or -NHCO-, a is an integer of 1 to 6,
A protein kinase degradation inducing compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. 8. The method of claim 7,
Formula L-2 is a structure of any one of Formulas L-3-1 to L-3-26 shown in the following table, wherein TL and ELL shown in Formulas L-3-1 to L-3-26 are Formula 1 represents the TL and ELL moieties as defined in, respectively, and k is an integer such that the minimum number of atoms covalently linked between the TL and ELL moieties is 1 to 30; Pharmaceutically acceptable salts of:

The method of claim 1,
A protein kinase degradation inducing compound selected from the group consisting of the following compounds, a stereoisomer or a pharmaceutically acceptable salt thereof:

ARK5, AURKA, AURKB, AURKC, CAMK1G, CAMK2A, CAMK2D, CAMKK1, CAMKK2, DYRK1A, EGFR(L858R/T790M), EGFR(T790M), EGFR(L858R), EGFR(Del19), EGFR(C797S), EGFR(Del19/C797S), EGFR(T790M/C797S), EGFR(Del19/T790M/C797S) ), EGFR(L858R/T790M/C797S), FLT3(D835H), FLT3(D835V), FLT3(D835Y), FLT3(ITD), FLT3(ITD,D835V), FLT3(ITD,F691L), FLT3(K663Q), FLT3 (N841I), FLT3 (R834Q), GRK1, GRK7, IKK-epsilon, IRAK3, JAK2 (JH1domain-catalytic), JAK3 (JH1domain-catalytic), KIT (A829P), KIT (D816H), KIT (D816V), MAP4K2 , MARK1, MLK1, MLK3, MST2, PDGFRB, PHKG1, PKN1, PKN2, PLK4, PRKCE, PRKG2, PYK2, QSK, RET(V804M), RSK2(Kin.Dom.1-N-terminal), RSK4(Kin.Dom) .1-N-terminal), SIK2, SNARK, TBK1, TNK1, TRKA, TRKB, or a pharmaceutical composition for the treatment or prevention of a YSK4-related disease. The pharmaceutical composition according to claim 10, wherein the disease is cancer or mastocytosis.

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