Compound with activity of degrading AKT kinase as well as preparation method and pharmaceutical application of compound

Abstract

The invention belongs to the field of medicinal chemistry, and discloses a compound with AKT kinase degradation activity and a preparation method and pharmaceutical application thereof, and the compound has a B-L-D structure as shown in a formula (I).

Description

Technical Field

[0001] The present invention relates to bivalent compounds (e.g., bifunctional small molecule compounds) that degrade and / or disrupt serine / threonine kinase AKT (also known as protein kinase B or PKB), compositions comprising one or more of said bivalent compounds, and methods of use thereof for the treatment of AKT-mediated diseases in a subject in need thereof. Background Art

[0002] The PI3K / AKT / mTOR pathway composed of phosphatidylinositol 3-kinase (PI3K), its downstream protein AKT (also known as protein kinase B, PKB), and mammalian target of rapamycin (mTOR) is a very important signal transduction pathway in cells and plays extremely important biological functions in processes such as cell growth, survival, proliferation, apoptosis, angiogenesis, and autophagy. Abnormal activation of this pathway can cause a series of diseases, including cancer, neuropathy, autoimmune diseases, and hematolymphatic system diseases.

[0003] AKT, a class of serine / threonine kinases, affects cell survival, growth, metabolism, proliferation, migration, and differentiation through numerous downstream effectors. More than 50% of human tumors exhibit overactivation of AKT, especially in prostate cancer, pancreatic cancer, bladder cancer, ovarian cancer, and breast cancer. Overactivation of AKT can lead to tumorigenesis, metastasis, and drug resistance.

[0004] AKT has three subtypes: AKT1, AKT2, and AKT3. As typical protein kinases, each subtype consists of an amino-terminal PH domain (Pleckstrin homology domain), a central kinase domain that binds ATP, and a carboxyl-terminal regulatory domain. Approximately 80% of the amino acid sequences of the three subtypes are homologous, with only relatively large variations in the region connecting the PH domain and the kinase domain.

[0005] At present, the targeted drugs for the PI3K / AKT / mTOR signaling pathway are mainly PI3K inhibitors and mTOR inhibitors, and AKT is at the core of this signal transduction pathway. Inhibiting AKT activity can avoid the serious side effects caused by inhibiting upstream PI3K, and also avoid the negative feedback mechanism caused by inhibiting downstream mTOR affecting the efficacy. Therefore, finding effective and selective AKT inhibitors is an important direction for the current research and development of tumor targeted drugs. CN101631778A discloses a class of cyclopentadienyl [D] pyrimidine derivatives, CN101578273A discloses a class of hydroxylated and methoxylated cyclopentadienyl [D] pyrimidine derivatives, CN101511842A discloses a class of dihydrofuranyl pyrimidine derivatives, and CN101970415A discloses a class of 5H-cyclopentadienyl [d] pyrimidine derivatives. These compounds have an IC of less than 10 μM for inhibiting AKT1. 50 .

[0006] Unlike conventional enzyme inhibitors that only inhibit the catalytic activity of the target enzyme, the AKT degradation / destruction compounds disclosed herein ("AKT degraders") bind to and induce the degradation of AKT, thereby eliminating any scaffolding function of AKT in addition to eliminating the enzymatic activity of AKT. The AKT degraders disclosed herein are bivalent compounds that include an AKT ligand conjugated to a degradation / destruction tag via a linker.

[0007] The AKT degraders disclosed herein provide a novel mechanism for treating AKT-mediated diseases. Specifically, in contrast to merely inhibiting AKT catalytic activity, it is expected that the ability of AKT degraders to target AKT for degradation overcomes resistance, regardless of whether a specific drug was used in prior treatment and regardless of whether resistance is caused by genetic mutation, amplification, or otherwise. Summary of the invention

[0008] The present invention generally relates to bivalent compounds (e.g., bifunctional compounds) that degrade and / or destroy AKT, and methods for treating AKT-mediated cancers (i.e., cancers that depend on AKT proteins; or cancers with elevated AKT levels or AKT activity relative to wild-type tissues of the same species and tissue type). Since AKT degraders / destructors have dual functions (enzyme inhibition plus protein degradation / destruction), the bivalent compounds disclosed in the present invention can be significantly more effective therapeutic agents than existing AKT inhibitors that inhibit the enzymatic activity of AKT but do not affect AKT protein levels. The present invention further provides methods for identifying AKT degraders / destructors as described herein.

[0009] More specifically, the present invention provides a bivalent compound comprising an AKT ligand conjugated to a degradation / destruction tag via a linker, having a structure as described in Formula I:

[0010] B—L—D(I)

[0011] L is selected from —Ak1—C1—Ak2—C2—Ak3— or —Ak4—Cy1—Ak5—Cy2—Ak6—;

[0012] wherein Ak1, Ak2, and Ak3 are independently selected from or a bond;

[0013] C1 and C2 are independently selected from C 1-12 alkyl, C 2-12 alkenyl, or C 2-12 alkynyl, or a bond, and C1 and C2 are not both a bond. The C 1-12 alkyl, C 2-12 alkenyl, or C 2-12 alkynyl is optionally substituted by one or more C 1-4 alkyl or halogen;

[0014] wherein Ak4, Ak5, and Ak6 are independently selected from C optionally substituted by one or more oxygen, C 1-4 alkyl, or halogen, C 1-12 alkyl, or a bond;

[0015] Cy1 and Cy2 are independently selected from a 3- to 12-membered heterocycle, a 3- to 12-membered cycloalkyl, a 6- to 10-membered aryl, or a bond, and Cy1 and Cy2 are not both a bond. The 3- to 12-membered heterocycle, 3- to 12-membered cycloalkyl, and 6- to 10-membered aryl are optionally substituted by one or more halogen or C 1-4 alkyl;

[0016] B is

[0017] wherein X is selected from C or N;

[0018] R 1 is halogen or hydrogen;

[0019] R 2 is selected from H, CN, C 1-6 alkyl, or C 1-6 alkoxy, wherein the C 1-6 alkyl or C 1-6 alkoxy is optionally substituted by halogen, OH, or CN;

[0020] G is a 5- to 12-membered saturated heterocycle containing 1 to 2 nitrogen atoms optionally substituted by C 1-4 alkyl;

[0021] R 3 is NH, or absent;

[0022] D is selected from the structures shown below:

[0023]

[0024] When B is L is not —Ak1—C1—Ak2—C2—Ak3—.

[0025] In some specific embodiments,

[0026] L is selected from alkyl groups of C 1-12 alkenyl groups of C 2-12 or alkynyl groups of C 2-12 wherein the alkyl groups of C 1-12 the alkenyl groups of C 2-12 or the alkynyl groups of C 2-12 are optionally substituted by one or more alkyl groups of C 1-4 or halogens,

[0027] or —Ak4—Cy1—Ak5—Cy2—Ak6;

[0028] wherein Ak4, Ak5, Ak6 are independently selected from alkyl groups of C 1-12 optionally substituted by one or more oxygen atoms or a bond;

[0029] Cy1, Cy2 are independently selected from 3- to 12-membered heterocycles optionally substituted by one or more halogens, alkyl groups of C 1-4 or a bond, and Cy1 and Cy2 are not both a bond at the same time.

[0030] In some more specific embodiments,

[0031] L is selected from alkyl groups of C 1-4 optionally substituted by one or more alkyl groups of C 1-12 or halogens, or alkynyl groups of C 2-12 ;

[0032] or —Ak4—Cy1—Ak5—Cy2—Ak6;

[0033] wherein Ak4, Ak5, Ak6 are independently selected from alkyl groups of C 1-10 substituted by one or more oxygen atoms or a bond; further, Ak4, Ak5, Ak6 are independently selected from alkyl groups of C 1-10 or a bond;

[0034] Cy1, Cy2 are independently selected from 4- to 9-membered heterocycles or a bond, and Cy1 and Cy2 are not both a bond at the same time.

[0035] Furthermore, Cy1, Cy2 are independently selected from 4- to 9-membered heterocycles containing 1 to 2 N atoms or a bond, and Cy1 and Cy2 are not both a bond at the same time.

[0036] Further, Cy1 and Cy2 are independently selected from azetidinyl, pyrrolidinyl, piperidine, piperazine, cyclopropyl-azetidinyl, cyclopropyl-pyrrolidinyl, cyclopropyl-piperidinyl, cyclopropyl-piperazinyl, cyclobutyl-azetidinyl, cyclobutyl-pyrrolidinyl, cyclobutyl-piperidinyl, cyclobutyl-piperazinyl, cyclopentyl-azetidinyl, cyclopentyl-pyrrolidinyl, cyclopentyl-piperidinyl, cyclopentyl-piperazinyl, cyclohexyl-azetidinyl, cyclohexyl-pyrrolidinyl, azetidinyl-azetidinyl, azetidinyl-pyrrolidinyl, azetidinyl-piperidinyl, azetidinyl-piperazinyl, pyrrolidinyl-azetidinyl, pyrrolidinyl-pyrrolidinyl, pyrrolidinyl-piperidinyl, pyrrolidinyl-piperazinyl, cyclobutyl-spiro-azetidinyl, cyclobutyl-spiro-pyrrolidinyl, cyclobutyl-spiro-piperidinyl, cyclopentyl-spiro-azetidinyl, cyclopentyl-spiro-pyrrolidinyl, cyclohexyl-spiro-azetidinyl, azetidinyl-spiro-azetidinyl, azetidinyl-spiro-pyrrolidinyl, azetidinyl-spiro-piperidinyl, pyrrolidinyl-spiro-azetidinyl, pyrrolidinyl-spiro-pyrrolidinyl, cyclohexyl-spiro-azetidinyl, cyclobutyl-spiro-piperazine, azetidinyl-spiro-piperazine or a bond, and Cy1 and Cy2 are not simultaneously a bond.

[0037] Further, Cy1 and Cy2 are independently selected from piperidine, piperazine or cyclobutyl-spiro-piperidinyl or a bond, and Cy1 and Cy2 are not simultaneously a bond.

[0038] Further, in some more typical embodiments,

[0039] L is selected from an optionally one or more methyl substituted C 1-12 alkyl or C 2-12 alkynyl, more preferably an optionally one or more methyl substituted C 8-12 alkyl or C 8-12 alkynyl, and further preferably C 8-12 alkyl or C 8-12 alkynyl.

[0040] Further, in some more typical embodiments,

[0041] L is selected from the following structures:

[0042]

[0043] where the left side of L is connected to B and the right side is connected to D.

[0044] Further, L is selected from the following structures:

[0045]

[0046] or

[0047] where the left side of L is connected to B and the right side is connected to D.

[0048] Further, L is In some specific embodiments,

[0049] B is

[0050] where X is selected from C or N;

[0051] R 1 is chlorine or hydrogen; R 2 is a C1-C6 alkyl group, where the C1-C6 alkyl group is optionally substituted by OH;

[0052] G is a 5-12 membered saturated heterocycle containing 1-2 nitrogen atoms optionally substituted by methyl;

[0053] R 3 is NH, or absent.

[0054] Further, R 2 is methyl, ethyl or —(CH2)2OH.

[0055] Further, R 2 is methyl or ethyl.

[0056] Further R 1 is chlorine.

[0057] Further, G is one of the following groups optionally substituted by methyl:

[0058] The single wavy line is the position where G is connected to the carbonyl group, and the double wavy line is the position where G is connected to the pyrimidine.

[0059] Further, G is selected from the following groups:

[0060]

[0061] Further, G is selected from the following groups:

[0062]

[0063] Further, in some more typical embodiments, B is selected from the following structures:

[0064] Further, in some more typical embodiments, B is selected from the following structures:

[0065] Further, in some more typical embodiments, B is selected from the following structures:

[0066] Further, in some more typical embodiments, B is selected from the following structures:

[0067] In some specific embodiments, D is selected from the following structures:

[0068]

[0069] Further, in some specific embodiments, D is selected from the following structures:

[0070]

[0071] Further, in some specific embodiments, D has the following structure

[0072]

[0073] In some more typical embodiments, the present invention provides a compound of the structure shown in II below or a pharmaceutically acceptable salt thereof:

[0074]

[0075] L1 is selected from —Ak1—C1—Ak2—C2—Ak3— or —Ak4—Cy1—Ak5—Cy2—Ak6—;

[0076] Wherein Ak1, Ak2, and Ak3 are independently selected from Or a bond;

[0077] C1 and C2 are independently selected from C 1-12 Alkyl of, C 2-12 Alkenyl of or C 2-12 Alkynyl of or a bond, and C1 and C2 are not both a bond, and the C 1-12 Alkyl of, C 2-12 Alkenyl of or C 2-12 Alkynyl of is optionally substituted by one or more C 1-4 Alkyl or halogen of;

[0078] Wherein Ak4, Ak5, and Ak6 are independently selected from C optionally substituted by one or more oxygen, C 1-4 Alkyl of or halogen substituted C 1-12 Alkyl of or a bond;

[0079] Cy1 and Cy2 are independently selected from 3- to 12-membered heterocycles, 3- to 12-membered cycloalkyls, 6- to 10-membered aryls or a bond, and Cy1 and Cy2 are not simultaneously a bond. The 3- to 12-membered heterocycles, 3- to 12-membered cycloalkyls, 6- to 10-membered aryls are optionally substituted by one or more halogens or C 1-4 alkyls substituted;

[0080] D1 is selected from the following structures:

[0081]

[0082] Furthermore,

[0083] L1 is —Ak1—C1—Ak2—C2—Ak3—, where Ak1, Ak2, Ak3 are independently selected from or a bond;

[0084] C1, C2 are independently selected from C 1-12 alkyls, C 2-12 alkenyls or C 2-12 alkynyls or a bond, and C1 and C2 are not simultaneously a bond. Among them, the C 1-12 alkyls, C 2-12 alkenyls or C 2-12 alkynyls are optionally substituted by one or more C 1-4 alkyls or halogens.

[0085] Furthermore,

[0086] L1 is selected from C 1-4 alkyls optionally substituted by one or more C 1-12 alkyls or halogens, C 2-12 alkenyls or C 2-12 alkynyls. Furthermore, preferably C 1-12 alkyls optionally substituted by one or more methyl groups, C 2-12 alkenyls or C 2-12 alkynyls. Further preferably C 8-12 alkyls or C 8-12 alkynyls optionally substituted by one or more methyl groups. Further preferably C 8-12 alkyls or C 8-12 alkynyls.

[0087] Furthermore, L1 is selected from the following structures:

[0088]

[0089] Furthermore, L1 is selected from the following structures:

[0090]

[0091] Further, D1 is selected from the following structures:

[0092]

[0093] Further, D1 is selected from the following structures:

[0094]

[0095] Further, in some specific embodiments, D1 is selected from the following structures:

[0096]

[0097] Further, in some specific embodiments, D1 has the following structure

[0098]

[0099] In some more typical embodiments, the present invention provides a compound of the structure shown in III below or a pharmaceutically acceptable salt thereof:

[0100]

[0101] L2 is selected from —Ak1—C1—Ak2—C2—Ak3— or —Ak4—Cy1—Ak5—Cy2—Ak6—;

[0102] Wherein Ak1, Ak2, and Ak3 are independently selected from Or a bond;

[0103] C1 and C2 are independently selected from C 1-12 Of alkyl, C 2-12 Of alkenyl or C 2-12 Of alkynyl or a bond, and C1 and C2 are not both a bond at the same time, wherein the C 1-12 Of alkyl, C 2-12 Of alkenyl or C 2-12 Of alkynyl is optionally substituted by one or more C 1-4 Of alkyl or halogen;

[0104] Wherein Ak4, Ak5, and Ak6 are independently selected from C optionally substituted by one or more oxygen, C 1-4 Of alkyl or halogen 1-12 Of alkyl or a bond;

[0105] Cy1 and Cy2 are independently selected from 3- to 12-membered heterocycles, 3- to 12-membered cycloalkyls, 6- to 10-membered aryls or a bond, and Cy1 and Cy2 are not both a bond at the same time, wherein the 3- to 12-membered heterocycles, 3- to 12-membered cycloalkyls, 6- to 10-membered aryls are optionally substituted by one or more halogens or C1-4 Alkyl substitution;

[0106] D2 is selected from the structures shown below:

[0107]

[0108] Furthermore,

[0109] L2 is —Ak1—C1—Ak2—C2—Ak3—, where Ak1, Ak2, and Ak3 are independently selected from or a bond;

[0110] C1 and C2 are independently selected from C 1-12 alkyl of, C 2-12 alkenyl of or C 2-12 alkynyl of or a bond, and C1 and C2 are not both a bond at the same time, where the C 1-12 alkyl of, C 2-12 alkenyl of or C 2-12 alkynyl of is optionally substituted by one or more C 1-4 alkyl of or halogen.

[0111] Furthermore,

[0112] L2 is selected from C 1-4 alkyl of optionally substituted by one or more C 1-12 alkyl of or halogen, C 2-12 alkenyl of or C 2-12 alkynyl of, furthermore, preferably C 1-12 alkyl of optionally substituted by one or more methyl groups, C 2-12 alkenyl of or C 2-12 alkynyl of, furthermore, more preferably C 8-12 alkyl of optionally substituted by one or more methyl groups or C 8-12 alkynyl of, furthermore, more preferably C 8-12 alkyl of or C 8-12 alkynyl of.

[0113] Furthermore, L2 is selected from the following structures:

[0114]

[0115] Furthermore, L2 is selected from the following structures:

[0116]

[0117] Furthermore, D2 is selected from the following structures:

[0118]

[0119] Further, D2 is selected from the following structures:

[0120]

[0121] Further, in some specific embodiments, D2 is selected from the following structures:

[0122]

[0123] Further, in some specific embodiments, D2 has the following structure

[0124]

[0125] In some more typical embodiments, the present invention provides a compound of the structure shown in IV below or a pharmaceutically acceptable salt thereof:

[0126]

[0127] Wherein, L3 is —Ak4—Cy1—Ak5—Cy2—Ak6—;

[0128] Wherein Ak4, Ak5, Ak6 are independently selected from C optionally substituted with one or more oxygen, C 1-4 alkyl or halogen-substituted C 1-12 alkyl or a bond;

[0129] Cy1, Cy2 are independently selected from a 3- to 12-membered heterocyclic ring, a 3- to 12-membered cycloalkyl, a 6- to 10-membered aryl or a bond, and Cy1 and Cy2 are not simultaneously a bond, wherein the 3- to 12-membered heterocyclic ring, 3- to 12-membered cycloalkyl, 6- to 10-membered aryl are optionally substituted with one or more halogen or C 1-4 alkyl;

[0130] D3 is selected from the structures shown below:

[0131]

[0132] Further, L3 is —Cy1—Ak6—;

[0133] Ak6 is C alkyl substituted with one or more oxygen; further preferably C 1-10 alkyl; further preferably C 3-10 alkyl;

[0134] Cy1 is a 4- to 9-membered heterocyclic ring, further preferably a 4- to 9-membered heterocyclic ring containing 1-2 Ns, further preferably piperidine, piperazine or cyclobutyl spiroazacyclohexyl, and further preferably piperazine.

[0135] Further, L3 is selected from the following structures:

[0136]

[0137]

[0138] Furthermore, D3 has the following structure:

[0139]

[0140] Furthermore, D3 has the following structure:

[0141]

[0142] In some more typical embodiments, the present invention provides a compound having the structure shown by V below or a pharmaceutically acceptable salt thereof:

[0143]

[0144] wherein, L4 is —Ak4—Cy1—Ak5—Cy2—Ak6—;

[0145] wherein Ak4, Ak5, and Ak6 are independently selected from C optionally substituted by one or more oxygen, C 1-4 alkyl or halogen-substituted C 1-12 alkyl or a bond;

[0146] Cy1 and Cy2 are independently selected from a 3- to 12-membered heterocycle, a 3- to 12-membered cycloalkyl, a 6- to 10-membered aryl or a bond, and Cy1 and Cy2 are not simultaneously a bond, wherein the 3- to 12-membered heterocycle, 3- to 12-membered cycloalkyl, 6- to 10-membered aryl are optionally substituted by one or more halogen or C 1-4 alkyl;

[0147] D4 is selected from the structures shown below:

[0148]

[0149] Furthermore, L4 is

[0150] —Ak4—Cy1—Ak5—Cy2—Ak6—;

[0151] wherein Ak4, Ak5, and Ak6 are independently selected from C substituted by one or more oxygen 1-10 alkyl or a bond;

[0152] Cy1 and Cy2 are independently selected from a 4- to 9-membered heterocycle or a bond, and Cy1 and Cy2 are not simultaneously a bond.

[0153] Furthermore, Cy1 and Cy2 are independently selected from a 4- to 9-membered heterocycle containing 1 to 2 N or a bond, and Cy1 and Cy2 are not simultaneously a bond.

[0154] Further, Cy1 and Cy2 are independently selected from azetidinyl, pyrrolidinyl, piperidine, piperazine, cyclopropyl-fused azetidinyl, cyclopropyl-fused pyrrolidinyl, cyclopropyl-fused piperidinyl, cyclopropyl-fused piperazinyl, cyclobutyl-fused azetidinyl, cyclobutyl-fused pyrrolidinyl, cyclobutyl-fused piperidinyl, cyclobutyl-fused piperazinyl, cyclopentyl-fused azetidinyl, cyclopentyl-fused pyrrolidinyl, cyclopentyl-fused piperidinyl, cyclopentyl-fused piperazinyl, cyclohexyl-fused azetidinyl, cyclohexyl-fused pyrrolidinyl, azetidinyl-fused azetidinyl, azetidinyl-fused pyrrolidinyl, azetidinyl-fused piperidinyl, azetidinyl-fused piperazinyl, pyrrolidinyl-fused azetidinyl, pyrrolidinyl-fused pyrrolidinyl, pyrrolidinyl-fused piperidinyl, pyrrolidinyl-fused piperazinyl, cyclobutyl-spiro-azetidinyl, cyclobutyl-spiro-pyrrolidinyl, cyclobutyl-spiro-piperidinyl, cyclopentyl-spiro-azetidinyl, cyclopentyl-spiro-pyrrolidinyl, cyclohexyl-spiro-azetidinyl, azetidinyl-spiro-azetidinyl, azetidinyl-spiro-pyrrolidinyl, azetidinyl-spiro-piperidinyl, pyrrolidinyl-spiro-azetidinyl, pyrrolidinyl-spiro-pyrrolidinyl, cyclohexyl-spiro-azetidinyl, cyclobutyl-spiro-piperidine, azetidinyl-spiro-piperidine, or a bond, provided that Cy1 and Cy2 are not both a bond at the same time.

[0155] Further, Cy1 and Cy2 are independently selected from piperidine, piperazine, cyclobutyl-spiro-piperidinyl, or a bond, provided that Cy1 and Cy2 are not both a bond at the same time.

[0156] Further, in some more typical embodiments,

[0157] L4 is selected from the following structures:

[0158]

[0159] where the left side of L is connected to B and the right side is connected to D4.

[0160] Further, in some more typical embodiments,

[0161] L4 is selected from the following structures:

[0162] where the left side of L is connected to B and the right side is connected to D4.

[0163] Further, D4 is selected from the following structures:

[0164]

[0165] Further, D4 is selected from the following structures:

[0166]

[0167] In some more typical embodiments, the present invention provides a compound of the structure shown in VI below or a pharmaceutically acceptable salt thereof:

[0168]

[0169] Wherein, L5 is —Ak4—Cy1—Ak5—Cy2—Ak6—;

[0170] Wherein Ak4, Ak5, and Ak6 are independently selected from C alkyl optionally substituted with one or more oxygen, C 1-4 alkyl or halogen-substituted C 1-12 alkyl or a bond;

[0171] Cy1 and Cy2 are independently selected from a 3- to 12-membered heterocycle, a 3- to 12-membered cycloalkyl, a 6- to 10-membered aryl, or a bond, and Cy1 and Cy2 are not simultaneously a bond, wherein the 3- to 12-membered heterocycle, 3- to 12-membered cycloalkyl, and 6- to 10-membered aryl are optionally substituted with one or more halogen or C 1-4 alkyl; D5 is selected from the structures shown below:

[0172]

[0173] Further, L5 is —Cy1—Ak6—;

[0174] Ak6 is C alkyl substituted with one or more oxygen 1-10 alkyl; more preferably C 3-10 alkyl;

[0175] Cy1 is a 4- to 9-membered heterocycle, further preferably a 4- to 9-membered heterocycle containing 1 to 2 N, more preferably piperidine, piperazine, or cyclobutylspiroazacyclohexyl, and even more preferably piperazine.

[0176] Further, L6 is selected from the following structures:

[0177]

[0178] In some more specific embodiments, the structure of the compound of formula I is as shown below:

[0179]

[0180]

[0181]

[0182]

[0183]

[0184] The bifunctional compound of formula I can be in the form of a free acid or free base, or a pharmaceutically acceptable salt. As used herein, the term "pharmaceutically acceptable salt" in context means a salt of the compound that does not destroy the biological activity or properties of the compound and is relatively non-toxic, i.e., the compound in salt form can be administered to a subject without causing an undesirable biological effect (e.g., dizziness or stomach upset) or an interaction in a harmful manner with any other ingredient contained in the composition. The term "pharmaceutically acceptable salt" means a product obtained by the reaction of a compound of the present invention with a suitable acid or base. Examples of pharmaceutically acceptable salts of the present invention include those derived from suitable inorganic bases such as salts of Li, Na, K, Ca, Mg, Fe, Cu, Al, Zn, and Mn. Examples of addition salts with inorganic acids are hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, galacturonate, formate, benzoate, glutamate, mesylate, esylate, benzenesulfonate, etc. Certain compounds of the present invention can form pharmaceutically acceptable salts with various organic bases such as arginine, guanidine, diethanolamine, or metformin.

[0185] In some embodiments, the compounds of the invention are isotope derivatives because they have at least one atom of the desired isotope substituted in an amount higher than the natural abundance of the isotope, i.e., are enriched. In some embodiments, the compound includes deuterium. Substitution with a heavier isotope, i.e., 2 H, can provide certain therapeutic advantages because of higher metabolic stability, e.g., increased in vivo half-life or reduced dosage requirements, and may thus be advantageous in certain circumstances.

[0186] The bifunctional compounds of formula I may have at least one chiral center and may thus be in the form of stereoisomers, as used herein, which encompasses all isomers of the individual compounds, which differ only in the orientation of their atoms in space. The term "stereoisomers" includes enantiomers (including the (R-) or (S-) configurations of the compound), mixtures of enantiomers (physical mixtures of enantiomers, racemates or racemic mixtures), geometric (cis / trans or E / Z) isomers of compounds, and isomers of compounds having more than one chiral center and not being mirror images of each other (diastereoisomers). The chiral centers of the compound may epimerize in vivo; thus, for these compounds, administration of the compound in the (R-) form is considered equivalent to administration of the compound in the (S-) form. Accordingly, the bifunctional compounds of formula I can be prepared and used in the form of the individual isomers and substantially free of other isomers, or in the form of mixtures of the various isomers, such as a racemic mixture of stereoisomers.

[0187] In some embodiments, the bifunctional compounds of formula I are isotope derivatives because they have at least one atom with a desired isotope substitution in an amount higher than the natural abundance of the isotope, i.e., they are enriched. In some embodiments, the compound includes deuterium. Substitution with a heavier isotope, i.e., 2H, may provide certain therapeutic advantages because of higher metabolic stability, e.g., increased in vivo half-life or reduced dosage requirements, and may thus be advantageous in certain cases.

[0188] Methods of synthesis

[0189] In some embodiments, the present invention relates to a method for preparing a bifunctional compound of formula I or a pharmaceutically acceptable salt or stereoisomer thereof. Generally, the bifunctional compound or its pharmaceutically acceptable salts or stereoisomers can be prepared by any method known to be suitable for preparing chemically related compounds. The synthetic schemes described in the specific examples of the various operations will better understand the compounds of the present invention, and these synthetic schemes illustrate non-limiting methods by which the compounds of the present invention can be prepared.

[0190] The present invention discloses a method for preparing a compound of formula I, which comprises:

[0191] Method 1:

[0192]

[0193] wherein R 1 、R 2 、G, L, D are as defined in general formula I, R 1a is selected from halogen, preferably bromine, and R 1b is methyl or hydrogen.

[0194] The compound of general formula (I-1A) and the compound of general formula (I-1B) are coupled or undergo nucleophilic substitution under palladium catalysis to obtain the compound of general formula (I-1C). The compound of general formula (I-1C) is oxidized to obtain the compound of general formula (I-1D). The compound of general formula (I-2D) and the compound of general formula (I-1E) undergo reductive amination to obtain the compound of general formula (I-1). Before the oxidation reaction, the compound of general formula (I-2C) can be reduced first to reduce the alkynyl group on the linker to an alkenyl or alkyl group.

[0195] Method 2:

[0196]

[0197] Among them, R 1 、R 2 、G, L, and D are defined as in general formula I, and R 2a is selected from halogen, preferably bromine.

[0198] The compound of general formula (I-2A) and the compound of general formula (I-2B) are coupled or undergo nucleophilic substitution under palladium catalysis to obtain the compound of general formula (I-2C). The compound of general formula (I-2C) is oxidized to obtain the compound of general formula (I-2D). The compound of general formula (I-2D) and the compound of general formula (I-2E) undergo reductive amination to obtain the compound of general formula (I-2). Before the oxidation reaction, the compound of general formula (I-2C) can be reduced first to reduce the alkynyl group on the linker to an alkenyl or alkyl group.

[0199] Method 3

[0200]

[0201] Among them, R 1 、R 2 、G, L, and D are defined as in general formula I,

[0202] The compound of general formula (I-2D) and the compound of general formula (I-2E) undergo reductive amination to obtain the compound of general formula (I).

[0203] Pharmaceutical composition

[0204] Another aspect of the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a bifunctional compound of formula I or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" as known in the art refers to a pharmaceutically acceptable material, composition or carrier suitable for administering the compounds of the present invention to a mammal. Suitable carriers may include, for example, liquids (aqueous and non-aqueous analogs, and combinations thereof), solids, encapsulating materials, gases, and combinations thereof (e.g., semi-solids), and compounds that play a role in carrying or transporting a gas from one organ or part of the body to another organ or part of the body. The carrier is "acceptable" in the sense that it is physiologically inert with respect to the other components of the formulation, compatible with the other components of the formulation, and not harmful to the subject or patient. Depending on the type of formulation, the composition may contain one or more pharmaceutically acceptable excipients.

[0205] Method of Use

[0206] The present invention relates to a method for treating a disease or disorder involving AKT activity dysfunction or dysregulation, which requires administering to a subject in need a therapeutically effective amount of a bifunctional compound of formula I or a pharmaceutically acceptable salt or stereoisomer thereof.

[0207] A disease or disorder can be said to be characterized or mediated by dysregulated AKT activity (e.g., elevated AKT levels or other functionally abnormal AKT relative to a non-pathological state). A "disease" is generally considered to be a state of health of a subject in which the subject is unable to maintain homeostasis and in which the subject's health continues to deteriorate if the disease is not improved. In contrast, a "disorder" of a subject is a state of health in which the subject is able to maintain homeostasis, but the subject's health is not as good as it would be in the absence of the disorder. A disorder does not necessarily lead to a further decline in the animal's health if left untreated.

[0208] The terms subject or patient as used herein include all members of the animal kingdom that are susceptible to or suffer from a designated disease or disorder. In some specific embodiments, the subject is a mammal, such as a human or non-human mammal. The method is also applicable to companion animals such as dogs and cats, as well as livestock such as cows, horses, sheep, goats, pigs, and other domesticated and wild animals. A subject "in need" of treatment according to the present invention may "have or be suspected of having" a particular disease or disorder, may have been positively diagnosed or otherwise presented with a sufficient number of risk factors or a sufficient number or combination of signs or symptoms such that a medical professional can diagnose or suspect that the subject has a disease or disorder. Thus, subjects having and suspected of having a particular disease or disorder are not necessarily two distinct groups.

[0209] In some specific examples, the compounds of formula I can be used to treat cell proliferative diseases and disorders (e.g., cancer or benign tumors). As used herein, the term "cell proliferative disease or disorder" refers to a condition characterized by a disorder or abnormality or both of cell growth, including non-cancerous conditions such as tumors, pre-cancerous conditions, and benign tumors.

[0210] Medical kit

[0211] The bifunctional compounds of the present invention and / or compositions containing these compounds can be assembled into kits or drug systems. The kits or drug systems according to this aspect of the present invention include a carrier or packaging, such as a box, carton, tube, etc., which has one or more containers, such as vials, tubes, ampoules, or bottles, which are tightly restricted to contain the compound of formula I or its pharmaceutical composition. The kits or drug systems of the present invention may also include printed instructions for using the compounds and compositions.

[0212] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter of this application belongs. As used in the specification and the appended claims, unless the context indicates otherwise, the following terms have the indicated meanings to facilitate understanding of the present invention.

[0213] The "compounds" described in the present invention include all stereoisomers and tautomers.

[0214] The compounds of the present invention can be asymmetric, for example, having one or more stereoisomers. Unless otherwise specified, all stereoisomers are included, such as enantiomers and diastereomers. The compounds of the present invention containing asymmetric carbon atoms can be isolated in optically pure form or in racemic form. The optically pure form can be resolved from the racemic mixture or synthesized by using chiral starting materials or chiral reagents. Racemates, diastereomers, and enantiomers are all included within the scope of the present invention.

[0215] The compounds of the present invention also include tautomeric forms. Tautomeric forms result from the exchange of a single bond with an adjacent double bond and are accompanied by the migration of a proton.

[0216] In the present invention, Ak1, Ak2, and Ak3 of L can be simultaneously bonds. When they are simultaneously bonds, L is "-C1-C2-". If C1 is also a bond, then L is "-C2-". Similarly, Ak4, Ak5, and Ak6 can also be simultaneously bonds, and L is "-Cy1-Cy2-". If Cy1 is also a bond, then L is "-Cy2-". If Ak1 and C1 are simultaneously bonds, then L is "-Ak2-C2-Ak3-". If Ak4, Cy1, and Ak5 are simultaneously bonds, then L is "-Cy2-Ak6-". That is, if adjacent ones are simultaneously bonds, it means that only one linking bond exists.

[0217] The structure of B in the present invention is as follows:

[0218] When R 3 is absent, it refers to the following structure: This structure is directly connected to L.

[0219] The term "optional" or "optionally" means that the subsequent described event or situation may or may not occur, and this description includes the occurrence and non-occurrence of the described event or situation.

[0220] Examples of the term "halogen" include fluorine atom, chlorine atom, bromine atom, iodine atom.

[0221] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain or branched saturated hydrocarbon groups, and the hydrocarbon group has the number of carbon atoms shown. For example, the term "C 1-12 alkyl" includes C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C10 alkyl, C11 alkyl, C12 alkyl. Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, n-hexyl, 2-hexyl, 3-hexyl, etc. It can be divalent, such as methylene, ethylene.

[0222] The term "alkenyl" refers to a straight-chain or branched hydrocarbon group containing at least one carbon-carbon double bond. Alkenyl includes groups with "cis" and "trans" orientations, or alternatively, "E" and "Z" orientations. For example, the term "C 8-12 alkenyl" includes C8 alkenyl, C9 alkenyl, C10 alkenyl, C11 alkenyl, C12 alkenyl. Examples include, but are not limited to, octene, (E)-non-2-ene, (E)-dec-2-ene, (E)-undec-2-ene, etc.

[0223] The term "alkynyl" refers to a straight-chain or branched hydrocarbon group containing at least one carbon-carbon triple bond. For example, the term "C 8-12 alkynyl" refers to a hydrocarbon group having 8 to 12 carbon atoms and containing at least one triple bond, and can be, but not limited to, octyne, (E)-non-2-yne, (E)-dec-2-yne, (E)-undec-2-yne, etc.

[0224] The term "alkoxy" refers to a group having an alkyl-O- structure, and the alkyl is a straight-chain or branched saturated monovalent hydrocarbon group. Non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentyloxy, n-hexyloxy.

[0225] The term "heterocyclic group" or "heterocycle" refers to a saturated or unsaturated non-aromatic ring system, which can be a 3- to 8-membered monocyclic ring, a 4- to 12-membered bicyclic ring or a 10- to 15-membered tricyclic ring, and contains 1 to 3 heteroatoms selected from N, O or S. Preferably, it is a 4- to 9-membered heterocyclic group. The N in the ring of the heterocyclic group can be selectively substituted and can be oxidized to various oxidation states. The heterocyclic group can be attached to a heteroatom or a carbon atom, and can be attached with a bridged ring or a spiro ring. Examples of monocyclic saturated heterocyclic groups are oxetanyl, azetidinyl, pyrrolidinyl, 2-oxo-pyrrolidin-3-yl, tetrahydrofuranyl, tetrahydro-thienyl, pyrazolidinyl, imidazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholin-4-yl, azepanyl, diazepanyl, homopiperazinyl or oxaazepanyl. Examples of bicyclic saturated heterocyclic groups are 8-aza-bicyclo[3.2.1]octyl, quinuclidinyl, 8-oxa-3-aza-bicyclo[3.2.1]octyl, 9-aza-bicyclo[3.3.1]nonyl. Examples of partially unsaturated heterocyclic groups are dihydrofuranyl, imidolinyl, tetrahydro-pyridyl or dihydropyranyl.

[0226] The term "cycloalkyl" refers to a monocyclic saturated hydrocarbon system without heteroatoms and without double bonds. Examples of the term "3-12 membered cycloalkyl" include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

[0227] The term "aryl" refers to a substituted or unsubstituted aromatic hydrocarbon group having a monocyclic or fused ring, and the number of ring atoms in the aromatic ring includes, but is not limited to, 6 to 18, 6 to 12 or 6 to 10 carbon atoms. The aryl ring can be fused to a saturated or unsaturated carbocyclic or heterocyclic ring, and the ring connected to the parent structure is the aryl ring. Non-limiting examples include benzene ring, naphthalene ring.

[0228] The term "bond" means that it is directly connected by a chemical single bond at this place.

[0229] Single wavy line Refers to the connection of chemical bonds.

[0230] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological activity of the free acid and base of a specific compound without biological adverse effects. For example, acid (including organic acid and inorganic acid) addition salts or base addition salts (including organic base and inorganic base).

[0231] The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compounds containing acid radicals or basic groups by conventional chemical methods. Generally, the preparation method of such salts is: in water or an organic solvent or a mixture of both, these compounds in the form of free acid or base are reacted with a stoichiometric amount of an appropriate base or acid to prepare.

[0232] The term "effective amount" or "therapeutically effective amount" refers to a sufficient amount of a drug or agent that is non-toxic but capable of achieving the desired effect.

[0233] The content of the present invention will be further understood by considering the following examples, which are intended to illustrate certain specific embodiments of the present invention but do not limit its scope of protection.

[0234] Synthesis section

[0235] Preparation of Intermediate 1 (R)-4-((1S,6R)-5-((S)-2-(4-chlorophenyl)-3-(piperazin-1-yl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one

[0236]

[0237] a) Preparation of (R)-4-((1S,6R)-5-((S)-3-(4-benzylpiperazin-1-yl)-2-(4-chlorophenyl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one

[0238] (R)-4-((1S,6R)-5-((S)-3-amino-2-(4-chlorophenyl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (500 mg), N-benzyl-2-bromo-N-(2-bromoethyl)ethan-1-amine (437 mg), N,N-diisopropylethylamine (293 mg), and ethanol (5 mL) were added to a reaction flask, and the mixture was stirred at room temperature overnight. After the reaction was completed, the resulting mixture was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to obtain 210 mg of the title compound.

[0239] b) Preparation of (R)-5-methyl-4-((1S,6R)-5-((S)-2-phenyl-3-(piperazin-1-yl)propanoyl)-2,5-diazabicyclo[4.1.0]heptan-2-yl)-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one

[0240] (R)-4-((1S,6R)-5-((S)-3-(4-Benzylpiperazin-1-yl)-2-(4-chlorophenyl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (210 mg), palladium / carbon (210 mg), palladium hydroxide / carbon (210 mg), 1,4-dioxane hydrochloride solution (4 M, 0.01 mL), and methanol (5 mL) were added to a reaction flask, and the mixture was stirred overnight under a hydrogen atmosphere at 50 °C. After the reaction was completed, the mixture was filtered, the filter cake was washed with methanol, the organic phase was collected and concentrated to obtain a crude product for the next step.

[0241] Example 1 Preparation of 3-(4-(10-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)amino)decyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0242]

[0243] a) Preparation of 3-(4-bromo-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0244] To 7-bromo-1-methyl-1,3-dihydro-2H-benzo[d]imidazol-2-one (5.0 g) and 3-bromopiperidine-2,6-dione (6.3 g) was added N,N-dimethylformamide (50 mL), and then sodium hydride (1.6 g) was added. The mixture was stirred at room temperature under a nitrogen atmosphere for 4 hours. After the reaction was completed, the resulting mixture was extracted with ethyl acetate (200 mL × 3). The combined organic layers were washed with brine (300 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with ethyl acetate / petroleum ether, to obtain 6.0 g of the title compound.

[0245] b) Preparation of 3-(4-(10-hydroxydec-1-yn-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0246] 3-(4-Bromo-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (1.0 g), 9-decyn-1-ol (912 mg), triethylamine (897 mg), copper(I) iodide (112 mg), bis(triphenylphosphine)palladium(II) chloride (207 mg), and N,N-dimethylformamide (10 mL) were added to a reaction flask. The mixture was stirred at 70 °C for 3 h under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was extracted with ethyl acetate (70 mL × 3). The combined organic layers were washed with brine (70 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with methanol / dichloromethane, to give 600 mg of the title compound.

[0247] c) Preparation of 3-(4-(10-Hydroxydecyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0248] Ethyl acetate (5 mL) was added to 3-(4-(10-hydroxydec-1-yn-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (300 mg) and palladium / carbon (60 mg), and the mixture was stirred at 50 °C under a hydrogen atmosphere for 15 h. The resulting mixture was filtered and the filter cake was washed with methanol (10 mL × 3). The filtrate was concentrated under reduced pressure. 200 mg of the title compound was obtained.

[0249] d) Preparation of 10-(1-(2,6-Dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)decanal

[0250] (1,1,1-Triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (206 mg) was added to dichloromethane (1 mL) of 3-(4-(10-hydroxydecyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (100 mg). The mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. After completion of the reaction, the mixture was diluted with saturated sodium bicarbonate (20 mL), and the resulting mixture was extracted with dichloromethane (30 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with ethyl acetate / petroleum ether, to give 80 mg of the title compound.

[0251] e) Preparation of 3-[4-(10-{[(2S)-2-(4-chlorophenyl)-3-[3-(5-ethyl-7-oxo-6,8-dihydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl]-3-oxopropyl]amino}dec-1-yn-1-yl)-3-methyl-2-oxo-1,3-benzoxazol-1-yl]piperidine-2,6-dione

[0252] Dissolve 10-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)decanal (80 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octane-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (110 mg) in 1,2-dichloroethane (2 mL) and stir at room temperature for 30 minutes. Then add sodium triacetoxyborohydride (103 mg) to the mixture and stir at room temperature for 1 hour under a nitrogen atmosphere. After completion of the reaction, concentrate the resulting mixture under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 37.8 mg of the title compound.

[0253] 11H NMR (400 MHz, DMSO-d6) δ 11.09 (s, 1H), 10.82 (d, J = 19.3 Hz, 1H), 8.09 (d, J = 23.4 Hz, 1H), 7.39 (d, J = 2.1 Hz, 2H), 7.36 (s, 2H), 7.03–6.90 (m, 2H), 6.85 (d, J = 3.5 Hz, 1H), 5.36 (dd, J = 12.5, 5.3 Hz, 1H), 5.33 (dd, J = 12.5, 5.3 Hz, 2H), 4.59 (t, J = 8.0 Hz, 1H) 4.57 (t, J = 8.0 Hz, 1H), 4.52 (s, 1H), 4.32–4.03 (m, 1H), 3.61 (d, J = 4.7 Hz, 3H), 3.55 (dd, J = 7.6, 2.4 Hz, 3H), 3.15 (dd, J = 20.9, 11.3 Hz, 3H), 2.87 (m, 1H), 2.85 (m, 1H), 2.84 (m, 1H), 2.82 (m, 2H), 2.80 (m, 1H), 2.71 (m, 1H), 2.70 (s, 1H), 2.68-2.62 (m, 3H), 2.33 (p, J = 1.8 Hz, 1H), 2.17–1.82 (m, 1H), 1.73 (d, J = 7.8 Hz, 1H), 1.65 (t, J = 8.8 Hz, 1H), 1.57 (d, J = 7.9 Hz, 2H), 1.33 (s, 5H), 1.23 (s, 6H), 1.18 (s, 3H), 0.92 (t, J = 7.0 Hz, 1H), 0.85 (t, J = 7.0 Hz, 1H).

[0254] LCMS m / z = 867 [M+1] +

[0255] Example 2 Preparation of 3-[4-(10-{[(2S)-2-(4-chlorophenyl)-3-[3-(5-ethyl-7-oxo-6,8-dihydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl]-3-oxopropyl]amino}dec-1-yn-1-yl)-3-methyl-2-oxo-1,3-benzoxazol-1-yl]piperidine-2,6-dione

[0256]

[0257] a) Preparation of 10-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)non-9-ynal

[0258] To a solution of 3-(4-(10-hydroxydec-1-yn-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (100 mg) in dichloromethane (1 mL) was added (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (206 mg). The mixture was stirred at room temperature under a nitrogen atmosphere for 2 h. After completion of the reaction, the mixture was diluted with saturated sodium bicarbonate (20 mL), and the resulting mixture was extracted with dichloromethane (30 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with ethyl acetate / petroleum ether, to give the title compound (80 mg).

[0259] b) Preparation of 3-[4-(10-{[(2S)-2-(4-chlorophenyl)-3-[3-(5-ethyl-7-oxo-6,8-dihydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl]-3-oxopropyl]amino}dec-1-yn-1-yl)-3-methyl-2-oxo-1,3-benzoxazol-1-yl]piperidine-2,6-dione

[0260] 10-(1-(2,6-Dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)non-9-ynal (80 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octane-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (110 mg) were dissolved in 1,2-dichloroethane (2 mL) and stirred at room temperature for 30 min. Then sodium triacetoxyborohydride (103 mg) was added to the mixture, and the mixture was stirred at room temperature under a nitrogen atmosphere for 1 h. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give the title compound (19.8 mg).

[0261] 11H NMR (400 MHz, DMSO-d6) δ 11.10 (s, 1H), 10.82 (d, J = 18.4 Hz, 1H), 8.09 (d, J = 25.9 Hz, 1H), 7.51–7.24 (m, 4H), 7.11 (d, J = 7.7 Hz, 1H), 7.06–7.01 (m, 1H), 6.98 (t, J = 7.8 Hz, 1H), 5.38 (dd, J = 12.7, 5.4 Hz, 1H), 4.72–4.36 (m, 4H), 4.23–4.13 (m, 1H), 3.62 (dd, J = 9.0, 3.5 Hz, 3H), 3.55 (d, J = 3.7 Hz, 2H), 3.17 (d, J = 12.4 Hz, 1H), 3.04–2.77 (m, 3H), 2.76–2.52 (m, 5H), 2.45 (d, J = 7.0 Hz, 4H), 2.09–1.97 (m, 2H), 1.76–1.50 (m, 5H), 1.38 (s, 4H), 1.25 (d, J = 13.1 Hz, 7H), 0.88 (dt, J = 28.4, 7.0 Hz, 3H).

[0262] LCMS m / z = 863 [M+1] +

[0263] Example 3 Preparation of 3-[4-(11-{[(2S)-2-(4-chlorophenyl)-3-[3-(5-ethyl-7-oxo-6,8-dihydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl]-3-oxopropyl]amino}undecyl)-3-methyl-2-oxo-1,3-benzoxazol-1-yl]piperidine-2,6-dione

[0264]

[0265] a) Preparation of ethyl (4,6-dichloropyrimidin-5-yl)glycinate

[0266] A solution of 4,6-dichloropyrimidin-5-amine (11 g) in tetrahydrofuran (120 mL) was cooled to 0 °C under a nitrogen atmosphere, and then sodium hydride (3.22 g) was slowly added over 30 minutes. Subsequently, ethyl bromoacetate (16.80 g) was added dropwise at 0 °C. Finally, tetrabutylammonium iodide (49.55 g) was added portionwise to the above mixture. The resulting mixture was stirred at room temperature overnight. After completion of the reaction, the reaction was quenched with water at 0 °C. The resulting mixture was extracted with ethyl acetate (200 mL × 3), and the combined organic layers were washed with brine (100 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with ethyl acetate / petroleum ether, to give 8.9 g of the title compound.

[0267] b) Preparation of 4-chloro-8-[(3,4-dimethoxyphenyl)methyl]-5,6-dihydropteridin-7-one

[0268] Ethyl (4,6-dichloropyrimidin-5-yl)glycinate (8.9 g), 3,4-dimethoxybenzylamine (6.55 g), triethylamine (7.92 g), and isopropanol (150 mL) were added to a reaction flask. The above mixture was stirred overnight at 80 °C. After the reaction was completed, it was cooled to room temperature. It was filtered, and the filter cake was washed with ethanol (20 mL × 3) to obtain 8.5 g of the title compound.

[0269] c) Preparation of 4-chloro-8-[(3,4-dimethoxyphenyl)methyl]-5-ethyl-6H-pteridin-7-one

[0270] The N,N-dimethylformamide (100 mL) of 4-chloro-8-[(3,4-dimethoxyphenyl)methyl]-5,6-dihydropteridin-7-one (7.3 g) was cooled to -10 °C, maintained under a nitrogen atmosphere, then iodoethane (5.1 g) was added, and finally sodium hydride (1.05 g) was slowly added, and the reaction continued at -10 °C for 3 hours. After the reaction was completed, it was quenched with an aqueous ammonium chloride solution. The resulting mixture was extracted with ethyl acetate (100 mL × 3), and the combined organic layers were washed with brine (50 mL × 3) and dried over anhydrous sodium sulfate. After filtering out the solid, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with ethyl acetate / petroleum ether (0 - 50%), to obtain 3.4 g of the title compound.

[0271] d) Preparation of 4-chloro-5-ethyl-5,8-dihydropteridin-7(6H)-one

[0272] A solution of 4-chloro-8-[(3,4-dimethoxyphenyl)methyl]-5-ethyl-6H-pteridin-7-one (3.3 g) in trifluoroacetic acid (30 mL) and n-butanol (10 mL) was stirred at 80 °C for 2 hours. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The pH of the mixture was adjusted to 9 with saturated sodium bicarbonate solution. The resulting mixture was extracted with ethyl acetate (50 mL × 3), and the combined organic layers were dried over anhydrous sodium sulfate. After filtering out the solid, the filtrate was concentrated under reduced pressure. The target compound 1.8 g was obtained.

[0273] e) Preparation of tert-butyl 3-(5-ethyl-7-oxo-6,8-dihydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

[0274] A solution of 4-chloro-5-ethyl-5,8-dihydropteridin-7(6H)-one (1.8 g), N,N-diisopropylethylamine (3.28 g) and tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (2.70 g) in acetonitrile (20 mL) was stirred at 100 °C for 8 hours. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The residue was adjusted to pH 5 with dilute hydrochloric acid. The resulting mixture was extracted with ethyl acetate (50 mL × 3), and the combined organic layers were dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. 2.8 g of the target compound was obtained.

[0275] f) Preparation of 4-{3,8-diazabicyclo[3.2.1]oct-3-yl}-5-ethyl-6,8-dihydropteridin-7-one

[0276] tert-Butyl 3-(5-ethyl-7-oxo-6,8-dihydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (2.8 g) was added to a 1,4-dioxane hydrochloride solution (4 M, 30 mL), and then stirred at room temperature for 3 hours. After completion of the reaction, it was concentrated under reduced pressure to obtain 3 g of the title compound.

[0277] g) Preparation of tert-butyl N-[(2S)-2-(4-chlorophenyl)-3-[3-(5-ethyl-7-oxo-6,8-dihydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl]-3-oxopropyl]carbamate

[0278] A solution of 4-{3,8-diazabicyclo[3.2.1]oct-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (3 g), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (5.93 g), N,N-diisopropylethylamine (4.03 g) and (S)-3-((tert-butoxycarbonyl)amino)-2-(4-chlorophenyl)propanoic acid (3.74 g) in N,N-dimethylformamide (40 mL) was stirred at room temperature for 3 hours. After completion of the reaction, the resulting mixture was extracted with ethyl acetate (50 mL × 3). The combined organic layers were washed with brine (30 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with methanol / dichloromethane (0% - 10%) to obtain 4.2 g of the title compound.

[0279] h) Preparation of 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]oct-3-yl}-5-ethyl-6,8-dihydropteridin-7-one

[0280] tert-Butyl N-[(2S)-2-(4-chlorophenyl)-3-[3-(5-ethyl-7-oxo-6,8-dihydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl]-3-oxopropyl]carbamate (4.2 g) was added to a 1,4-dioxane hydrochloride solution (4 M, 40 mL), and then stirred at room temperature for 2 hours. After the reaction was completed, it was concentrated under reduced pressure, and the pH of the residue was adjusted to 9 with saturated sodium bicarbonate solution. The resulting mixture was extracted with ethyl acetate (100 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with methanol / dichloromethane (0% - 10%) to give 3.4 g of the title compound.

[0281] i) Preparation of 3-[4-(11-{[(2S)-2-(4-chlorophenyl)-3-[3-(5-ethyl-7-oxo-6,8-dihydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl]-3-oxopropyl]amino}undecyl)-3-methyl-2-oxo-1,3-benzoxazol-1-yl]piperidine-2,6-dione

[0282] 11-(1-(2,6-Dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)undecanal (50 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octan-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (55 mg) were dissolved in 1,2-dichloroethane (2 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (49.5 mg) was added to the mixture, and it was stirred at room temperature for 1 hour under a nitrogen atmosphere. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 8.4 mg of the title compound.

[0283] 11H NMR (400 MHz, DMSO-d6) δ 11.08 (s, 1H), 10.81 (d, J = 18.9 Hz, 1H), 8.08 (d, J = 23.1 Hz, 1H), 7.47–7.28 (m, 4H), 7.03–6.86 (m, 2H), 6.85 (dd, J = 5.4, 3.5 Hz, 1H), 5.36 (dd, J = 12.6, 5.4 Hz, 1H), 4.54 (dt, J = 29.8, 8.3 Hz, 3H), 4.20–4.05 (m, 1H), 3.61 (d, J = 4.5 Hz, 1H), 3.56 (d, J = 3.5 Hz, 1H), 3.54 (s, 3H), 3.30–3.23 (m, 2H), 3.13 (dt, J = 21.0, 10.7 Hz, 2H), 3.02–2.70 (m, 5H), 2.76–2.54 (m, 4H), 2.05–1.84 (m, 2H), 1.75 (dd, J = 11.3, 7.2 Hz, 2H), 1.62 (dd, J = 35.2, 7.7 Hz, 3H), 1.33 (s, 6H), 1.20 (d, J = 19.5 Hz, 11H), 0.88 (dt, J = 29.3, 7.0 Hz, 3H).

[0284] LCMS m / z = 881 [M+1] +

[0285] Example 4 Preparation of 3-(4-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydro-pteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0286]

[0287] a) Preparation of 3-(4-(11-hydroxyundec-1-yn-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione 3-(4-Bromo-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (1.0 g), undec-10-yn-1-ol (950 mg), triethylamine (897 mg), copper(I) iodide (112 mg), bis(triphenylphosphine)palladium(II) chloride (207 mg) and N,N-dimethylformamide (10 mL) were added to a reaction flask. The mixture was stirred at 70 °C for 3 hours under a nitrogen atmosphere. After the reaction was completed, the resulting mixture was extracted with ethyl acetate (70 mL × 3). The combined organic layers were washed with brine (70 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with methanol / dichloromethane (0% - 10%) to give 500 mg of the title compound.

[0288] b) Preparation of 11-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)undec-10-ynal

[0289] (1,1,1-Triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (199 mg) was added to a dichloromethane (1 mL) solution of 3-(4-(11-hydroxyundec-1-yn-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (100 mg). The mixture was stirred at room temperature for 2 hours under a nitrogen atmosphere. After the reaction was completed, the mixture was diluted with saturated sodium bicarbonate (20 mL), and the resulting mixture was then extracted with dichloromethane (30 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with ethyl acetate / petroleum ether (10% - 70%) to give 50 mg of the title compound.

[0290] c) Preparation of 3-[4-(11-([(2S)-2-(4-chlorophenyl)-3-[3-(5-ethyl-7-oxo-6,8-dihydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl]-3-oxopropyl]amino)undec-1-yn-1-yl)-3-methyl-2-oxo-1,3-benzoxazol-1-yl]piperidine-2,6-dione

[0291] 11-(1-(2,6-Dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)undec-10-ynal (50 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octan-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (61 mg) were dissolved in 1,2-dichloroethane (2 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (62.5 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 10.4 mg of the title compound.

[0292] 1 H NMR (400 MHz, DMSO-d6) δ 11.10 (s, 1H), 10.84–10.79 (d, 1H), 8.12 (s, 1H), 8.05–7.10 (m, 4H), 7.04 (d, J = 7.5 Hz, 1H), 6.98 (t, J = 7.8 Hz, 2H), 5.41–5.33 (m, 1H), 4.58–4.47 (m, 4H), 4.22 (d, J = 12.4 Hz, 1H), 4.16 (s, 3H), 3.65–3.60 (m, 1H), 3.60–3.58 (m, 1H), 3.56 (m, 1H), 2.89–2.86 (m, 3H), 2.86–2.70 (m, 5H), 2.37–2.31 (s, 2H), 2.11–2.05 (m, 1H), 2.02 (s, 2H), 1.73 (s, 3H), 1.66 (d, J = 7.0 Hz, 3H), 1.56–1.37 (m, 4H), 1.22 (d, J = 14.7 Hz, 9H), 0.92–0.85 (m, 3H).

[0293] LCMS m / z = 877 [M+1] +

[0294] Example 5 Preparation of 5-(11-(((S)-2-(4-chlorophenyl)-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl-3-oxopropyl)amino)undecyl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0295]

[0296] a) Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(11-hydroxyundec-1-yn-1-yl)isoindole-1,3-dione

[0297] 5-Bromo-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (1 g), undec-10-yn-1-ol (1 g), bis(triphenylphosphine)palladium(II) dichloride (0.21 g), and copper(I) iodide (0.11 g) were added to triethylamine (5 mL) and N,N-dimethylformamide (5 mL), and the reaction was carried out at 70 °C for 3 hours under a nitrogen atmosphere. After the reaction was completed, water (100 mL) was added to quench the reaction, and then the mixture was extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with brine (100 mL × 2) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with methanol / dichloromethane (0% - 10%) to give 900 mg of the title compound.

[0298] b) Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(11-hydroxyundecyl)isoindole-1,3-dione

[0299] Methanol (3 mL) was added to 2-(2,6-dioxopiperidin-3-yl)-5-(11-hydroxyundec-1-yn-1-yl)isoindole-1,3-dione (300 mg) and palladium / carbon (60 mg), and the mixture was stirred at 50 °C under a hydrogen atmosphere for 24 hours. The resulting mixture was filtered and the filter cake was washed with methanol (10 mL × 3). The filtrate was concentrated under reduced pressure to give 200 mg of the title compound.

[0300] c) Preparation of 11-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]undecanal

[0301] (1,1,1-Triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (398 mg) was added to a solution of 2-(2,6-dioxopiperidin-3-yl)-5-(11-hydroxyundecyl)isoindole-1,3-dione (200 mg) in dichloromethane (5 mL). The mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. After the reaction was completed, the mixture was quenched with water (100 mL), and then extracted with ethyl acetate (50 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with methanol / dichloromethane (0% - 10%) to give 70 mg of the title compound.

[0302] d) Preparation of 5-(11-(((S)-2-(4-chlorophenyl)-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undecyl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0303] Dissolve 11-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl]undecanal (100 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octan-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (110 mg) in 1,2-dichloroethane (1 mL) and stir at room temperature for 30 minutes. Then add sodium triacetoxyborohydride (159.5 mg) to the mixture and stir at room temperature for 1 hour under a nitrogen atmosphere. After completion of the reaction, concentrate the resulting mixture under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 13.2 mg of the title compound.

[0304] 1 H NMR (400 MHz, DMSO-d6) δ 10.83 (d, J = 19.4 Hz, 1H), 8.19 (s, 1H), 8.08 (d, J = 23.8 Hz, 1H), 7.83 (d, J = 7.7 Hz, 1H), 7.76 (s, 1H), 7.70 (d, J = 7.7 Hz, 1H), 7.38 (d, J = 13.0 Hz, 4H), 5.13 (dd, J = 13.0, 5.4 Hz, 1H), 4.60 (d, J = 14.2 Hz, 2H), 4.51 (s, 2H), 4.22–4.12 (m, 2H), 3.61 (d, J = 4.5 Hz, 1H), 3.56 (d, J = 3.7 Hz, 1H), 3.21–3.11 (m, 2H), 3.01–2.95 (m, 1H), 2.97–2.84 (m, 4H), 2.84–2.78 (m, 1H), 2.81–2.73 (m, 1H), 2.77–2.68 (m, 2H), 2.10–1.99 (m, 2H), 1.95–1.82 (m, 1H), 1.79–1.70 (m, 2H), 1.70–1.54 (m, 3H), 1.37–1.13 (m, 16H), 0.863–0.828 (m, 3H).

[0305] LCMS m / z = 881 [M+1] +

[0306] Preparation of Example 6 3-(4-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)amino)undecyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0307]

[0308] 11-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)undecanal (70 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octane-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (80.5 mg) were dissolved in 1,2-dichloroethane (2 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (72 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 9.1 mg of the title compound. 11H NMR (400 MHz, DMSO-d6) δ 10.83 (d, J = 18.9 Hz, 1H), 8.08 (d, J = 22.9 Hz, 1H), 7.56 (dd, J = 4.9, 3.7 Hz, 1H), 7.48–7.42 (m, 2H), 7.41–7.31 (m, 4H), 5.13 (dd, J = 13.2, 5.1 Hz, 1H), 4.70–4.46 (m, 3H), 4.43 (s, 1H), 4.29 (d, J = 17.1 Hz, 1H), 4.17 (ddd, J = 21.8, 19.0, 9.8 Hz, 1H), 3.61 (d, J = 4.4 Hz, 1H), 3.56 (d, J = 3.6 Hz, 1H), 3.15 (p, J = 10.0, 9.1 Hz, 1H), 2.96 (dd, J = 11.9, 5.9 Hz, 1H), 2.93–2.86 (m, 1H), 2.81 (q, J = 7.0 Hz, 1H), 2.73–2.65 (m, 1H), 2.62 (d, J = 5.2 Hz, 3H), 2.57 (d, J = 2.3 Hz, 3H), 2.41 (dd, J = 13.2, 9.0 Hz, 1H), 2.33 (p, J = 1.9 Hz, 1H), 2.11–1.94 (m, 1H), 1.90 (d, J = 7.1 Hz, 3H), 1.80 (m, 2H), 1.72–1.57 (m, 3H), 1.29 (d, J = 4.8 Hz, 6H), 1.19 (d, J = 19.3 Hz, 10H), 0.88 (dt, J = 28.7, 7.0 Hz, 3H).

[0309] LCMS m / z = 866 [M+1] +

[0310] Example 7 Preparation of 3-(5-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)amino)undecyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0311]

[0312] a) Preparation of 3-(5-(11-hydroxyundec-1-yn-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0313] 3-(5-Bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (1 g), undec-10-yn-1-ol (780 mg), copper(I) iodide (120 mg), triethylamine (940 mg), and bis(triphenylphosphine)palladium(II) chloride (250 mg) were added to N,N-dimethylformamide (10 mL). After purging with nitrogen, the reaction was carried out at 70 °C for 3 h. After completion of the reaction, the mixture was extracted with ethyl acetate (70 mL × 3). The combined organic phases were washed with saturated brine (200 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: methanol / dichloromethane) to give 1.1 g of the title compound.

[0314] b) Preparation of 3-(5-(11-hydroxyundecyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0315] 3-(5-(11-Hydroxyundec-1-yn-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (300 mg) and palladium / carbon (60 mg) were added to methanol (3 mL). The mixture was stirred at 50 °C for 15 h under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with methanol (10 mL × 3). The filtrate was concentrated under reduced pressure to give 270 mg of the title compound.

[0316] c) Preparation of 11-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)undecanal

[0317] 3-(5-(11-Hydroxyundecyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (100 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (204 mg) were added to dichloromethane (2 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 h. After the reaction, dichloromethane (30 mL) was added, and the organic phase was washed with saturated brine (50 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 35 mg of the title compound.

[0318] d) Preparation of 3-(5-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)amino)undecyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0319] 11-(2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)undecanal (50 mg) and 4-{8-[(2S)-3-Amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octan-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (68 mg) were dissolved in 1,2-dichloroethane (2 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (64 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 2.1 mg of the title compound.

[0320] 1 H NMR (400 MHz, DMSO-d6) δ 11.05 - 10.74 (m, 2H), 8.08 (m, 1H), 7.62 (d, J = 7.7 Hz, 1H), 7.46 - 7.28 (m, 6H), 5.09 (dd, J = 13.3, 5.1 Hz, 1H), 4.64 - 4.46 (m, 4H), 4.41 (d, J = 17.2 Hz, 1H), 4.31 - 4.24 (m, 1H), 4.23 - 4.10 (m, 2H), 3.58 (m, 2H), 3.01 - 2.86 (m, 1H), 2.86 - 2.76 (m, 2H), 2.72 - 2.65 (m, 1H), 2.62 - 2.57 (m, 3H), 2.56 - 2.53 (m, 1H), 2.43 - 2.28 (m, 3H), 2.12 - 1.86 (m, 1H), 1.77 - 1.71 (m, 1H), 1.67 - 1.52 (m, 4H), 1.41 - 1.06 (m, 18H), 0.80 - 0.96 (m, 3H).

[0321] LCMS m / z = 866 [M+1] +

[0322] Example 8 Preparation of 3-(5-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undecyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0323]

[0324] a) Preparation of 3-(5-(11-hydroxyundec-1-yn-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione 3-(5-Bromo-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (700 mg), undec-10-yn-1-ol (418 mg), copper(I) iodide (79 mg), triethylamine (419 mg), and bis(triphenylphosphine)palladium(II) chloride (145 mg) were added to N,N-dimethylformamide (7 mL). After purging with nitrogen, the reaction was carried out at 70 °C for 3 hours. After completion of the reaction, the mixture was extracted with ethyl acetate (70 mL × 3). The combined organic phases were washed with saturated brine (200 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: methanol / dichloromethane) to give 600 mg of the title compound.

[0325] b) Preparation of 3-(5-(11-hydroxyundecyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0326] 3-(5-(11-Hydroxyundec-1-yn-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (300 mg) and palladium / carbon (60 mg) were added to methanol (10 mL). The mixture was stirred overnight at 50 °C under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with methanol (10 mL × 3). The filtrate was concentrated under reduced pressure to give 250 mg of the title compound.

[0327] c) Preparation of 11-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)undecanal

[0328] 3-(5-(11-Hydroxyundecyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (100 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (197.5 mg) were added to dichloromethane (1 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 hours. After the reaction, dichloromethane (30 mL) was added, and the organic phase was washed with saturated brine (50 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 60 mg of the title compound.

[0329] d) Preparation of 3-(5-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undecyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0330] 11-(1-(2,6-Dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)undecanal (50 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octan-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (82.5 mg) were dissolved in 1,2-dichloroethane (1 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (50 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 5.4 mg of the title compound.

[0331] 1 H NMR (400 MHz, DMSO-d6) δ 10.81 (d, J = 19.2 Hz, 2H), 8.08 (m, 1H), 7.39 (d, J = 3.1 Hz, 2H), 7.36 (s, 2H), 7.06 - 6.92 (m, 2H), 6.85 (d, J = 8.0 Hz, 1H), 5.33 (dd, J = 12.7, 5.4 Hz, 1H), 4.62 - 4.46 (m, 3H), 4.27 - 4.01 (m, 2H), 3.58 (m, 2H), 3.26 - 3.04 (m, 2H), 3.00 - 2.84 (m, 2H), 2.80 (t, J = 7.1 Hz, 1H), 2.72 (d, J = 13.0 Hz, 2H), 2.67 (p, J = 1.9 Hz, 2H), 2.61 (d, J = 18.6 Hz, 3H), 2.14 - 1.94 (m, 2H), 1.94 - 1.62 (m, 4H), 1.57 (s, 3H), 1.23 (m, 18H), 0.88 (m, 3H).

[0332] LCMS m / z = 881 [M + 1] +

[0333] Preparation of 5-(11-(((S)-2-(4-chlorophenyl)-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0334] a) Preparation of 11-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)undec-1-ynal

[0335] 2-(2,6-Dioxopiperidin-3-yl)-5-(11-hydroxyundec-1-yn-1-yl)isoindoline-1,3-dione (170 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (340 mg) were added to dichloromethane (2 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 hours. After the reaction, dichloromethane (30 mL) was added. The organic phase was washed with saturated brine (50 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to obtain 106 mg of the title compound.

[0336] b) Preparation of 5-(11-(((S)-2-(4-chlorophenyl)-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0337] 11-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)undec-1-ynal (106 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octan-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (118 mg) were dissolved in 1,2-dichloroethane (2 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (160 mg) was added to the mixture, and the mixture was stirred at room temperature under a nitrogen atmosphere for 1 hour. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 11.6 mg of the title compound.

[0338] 11H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 10.83 (d, J = 20.4 Hz, 1H), 8.09 (d, J = 26.9 Hz, 1H), 7.89 (d, J = 8.1 Hz, 1H), 7.84 (dd, J = 5.5, 1.6 Hz, 2H), 7.48–7.30 (m, 4H), 5.15 (dd, J = 12.8, 5.4 Hz, 1H), 4.53 (m, 4H), 4.28–4.10 (m, 2H), 3.61 (d, J = 5.2 Hz, 1H), 3.56 (d, J = 4.0 Hz, 1H), 2.92 (m, 4H), 2.81 (d, J = 7.0 Hz, 3H), 2.67 (dt, J = 3.7, 1.9 Hz, 3H), 2.62 (s, 2H), 2.56 (d, J = 13.0 Hz, 3H), 2.12–1.91 (m, 2H), 1.73 (s, 1H), 1.56 (d, J = 7.7 Hz, 3H), 1.37 (s, 3H), 1.23 (d, J = 14.7 Hz, 8H), 0.92 (t, J = 7.0 Hz, 1H), 0.84 (t, J = 7.0 Hz, 1H). LCMS m / z = 876 [M+1] +

[0339] Example 10 Preparation of 3-(4-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0340]

[0341] 11-(2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindol-4-yl)undec-10-ynal (70 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octane-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (80 mg) were dissolved in 1,2-dichloroethane (2 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (72 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 6.5 mg of the title compound. 11H NMR (400 MHz, DMSO-d6) δ 10.83 (d, J = 19.2 Hz, 1H), 8.09 (d, J = 24.7 Hz, 1H), 7.70 (d, J = 7.5 Hz, 1H), 7.62 (d, J = 7.5 Hz, 1H), 7.51 (t, J = 7.6 Hz, 1H), 7.38 (d, J = 12.4 Hz, 4H), 5.15 (dd, J = 13.4, 5.1 Hz, 1H), 4.64–4.43 (m, 3H), 4.42 (s, 1H), 4.31 (s, 1H), 4.29–4.20 (m, 1H), 4.19–4.08 (m, 1H), 3.61 (d, J = 4.5 Hz, 1H), 3.56 (d, J = 3.7 Hz, 1H), 3.13 (dt, J = 20.8, 10.8 Hz, 1H), 2.93 (ddd, J = 22.7, 12.5, 5.7 Hz, 1H), 2.81 (q, J = 7.1 Hz, 1H), 2.73–2.63 (m, 1H), 2.57 (d, J = 3.6 Hz, 3H), 2.44 (d, J = 7.2 Hz, 2H), 2.43–2.37 (m, 1H), 2.33 (p, J = 1.9 Hz, 2H), 2.17 (d, J = 12.5 Hz, 1H), 2.12–1.96 (m, 1H), 1.90 (d, J = 7.2 Hz, 1H), 1.73 (d, J = 7.9 Hz, 1H), 1.66 (d, J = 8.6 Hz, 2H), 1.54 (dq, J = 14.0, 7.0 Hz, 1H), 1.32 (s, 2H), 1.23 (s, 3H), 1.19 (s, 3H), 0.92 (t, J = 7.1 Hz, 6H), 0.84 (t, J = 7.0 Hz, 3H).

[0342] LCMS m / z = 862 [M+1] +

[0343] Example 11 Preparation of 3-(5-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0344]

[0345] a) Preparation of 11-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)undec-10-ynal

[0346] 3-(5-(11-Hydroxyundec-1-yn-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (100 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (206 mg) were added to dichloromethane (2 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 hours. After the reaction, dichloromethane (30 mL) was added, and the organic phase was washed with saturated brine (50 mL×3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 80 mg of the title compound.

[0347] b) Preparation of 3-(5-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0348] 11-(2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)undec-10-ynal (80 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octane-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (138 mg) were dissolved in 1,2-dichloroethane (2 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (124 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 5.1 mg of the title compound. 11H NMR (400 MHz, DMSO-d6) δ 10.85 (s, 1H), 8.09 (d, J = 25.7 Hz, 1H), 7.68 (d, J = 7.9 Hz, 1H), 7.61 (s, 1H), 7.49 (d, J = 8.0 Hz, 1H), 7.42 - 7.33 (d, J = 11.1 Hz, 4H), 5.11 (dd, J = 13.3, 5.0 Hz, 1H), 4.64 - 4.55 (m, 2H), 4.54 - 4.88 (m, 2H), 4.44 (d, J = 17.5 Hz, 1H), 4.31 (d, J = 17.4 Hz, 1H), 4.27 - 4.16 (m, 1H), 4.18 - 4.07 (m, 1H), 3.61 (d, J = 5.1 Hz, 1H), 3.56 (d, J = 3.9 Hz, 1H), 3.20 - 3.03 (m, 1H), 3.00 - 2.89 (m, 1H), 2.88 - 2.77 (m, 1H), 2.72 - 2.66 (m, 2H), 2.64 - 2.58 (m, 2H), 2.47 - 2.43 (m, 1H), 2.42 – 2.39 (m, 1H), 2.38 - 2.31 (m, 1H), 2.12 - 2.06 (m, 1H), 2.04 - 1.95 (m, 1H), 1.93 - 1.85 (m, 1H), 1.79 - 1.71 (m, 1H), 1.68 - 1.63 (m, 1H), 1.60 - 1.49 (m, 2H), 1.45 - 1.32 (m, 5H), 1.29 - 1.15 (m, 9H), 0.92 (t, J = 7.0 Hz, 2H), 0.85 (t, J = 6.9 Hz, 2H).

[0349] LCMS m / z = 862 [M+1] +

[0350] Example 12 Preparation of 4-(11-(((S)-2-(4-chlorophenyl)-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undecyl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0351]

[0352] a) Preparation of 2-(2,6-dioxopiperidin-3-yl)-4-(11-hydroxyundec-1-yn-1-yl)isoindole-1,3-dione

[0353] 4-Bromo-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (1 g), undec-10-yn-1-ol (1 g), bis(triphenylphosphine)palladium(II) dichloride (0.21 g) and copper(I) iodide (0.11 g) were added to triethylamine (5 mL) and N,N-dimethylformamide (5 mL), and the reaction was carried out at 70 °C for 3 h under a nitrogen atmosphere. After the reaction was completed, water (100 mL) was added to quench the reaction, and then the mixture was extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with brine (100 mL × 2) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (mobile phase: methanol / dichloromethane) to give 900 mg of the title compound.

[0354] b) Preparation of 2-(2,6-dioxopiperidin-3-yl)-4-(11-hydroxyundecyl)isoindoline-1,3-dione

[0355] Methanol (7 mL) was added to 2-(2,6-dioxopiperidin-3-yl)-4-(11-hydroxyundec-1-yn-1-yl)isoindoline-1,3-dione (400 mg) and palladium / carbon (200 mg), and the mixture was stirred overnight at 50 °C under a hydrogen atmosphere. The resulting mixture was filtered and the cake was washed with methanol (10 mL × 3). The filtrate was concentrated under reduced pressure. 300 mg of the title compound was obtained.

[0356] c) Preparation of 11-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]undecanal

[0357] (1,1,1-Triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (200 mg) was added to dichloromethane (2 mL) of 2-(2,6-dioxopiperidin-3-yl)-4-(11-hydroxyundecyl)isoindoline-1,3-dione (200 mg). The mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. After the reaction was completed, the mixture was quenched with water (100 mL), and then extracted with ethyl acetate (50 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether = 0%-70%) to give 80 mg of the title compound.

[0358] d) Preparation of 4-(11-(((S)-2-(4-chlorophenyl)-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undecyl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0359] 11-[2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]undecanal (80 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octan-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (106 mg) were dissolved in 1,2-dichloroethane (1 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (79.5 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 25.7 mg of the title compound.

[0360] 1 H NMR (400 MHz, DMSO-d6) δ 10.81 (d, J = 17.3 Hz, 1H), 8.28 (s, 1H), 8.08 (d, J = 22.6 Hz, 1H), 7.81 - 7.72 (m, 2H), 7.69 (dd, J = 6.9, 1.9 Hz, 1H), 7.49 - 7.31 (m, 4H), 5.12 (dd, J = 12.9, 5.4 Hz, 1H), 4.67 - 4.43 (m, 4H), 4.28 - 4.12 (m, 2H), 3.61 (d, J = 4.3 Hz, 1H), 3.56 (d, J = 3.6 Hz, 1H), 3.17 (d, J = 11.5 Hz, 2H), 3.14 - 3.06 (m, 2H), 3.05 - 2.99 (m, 2H), 2.89 (dd, J = 5.5, 3.3 Hz, 1H), 2.83 (dd, J = 14.8, 6.7 Hz, 2H), 2.75 - 2.66 (m, 2H) 2.62 (d, J = 4.5 Hz, 2H), 2.13 - 1.99 (m, 2H), 1.90 (d, J = 7.1 Hz, 1H), 1.73 (d, J = 8.0 Hz, 2H), 1.63 (dd, J = 29.2, 8.3 Hz, 4H), 1.30 (s, 6H), 1.19 (d, J = 18.2 Hz, 8H), 0.88 (dt, J = 28.5, 7.0 Hz, 3H).

[0361] LCMS m / z = 880 [M+1] +

[0362] Preparation of 1-(6-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undecyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0363]

[0364] a) Preparation of 1-(6-(11-hydroxyundec-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0365] 1-(6-Bromo-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (1 g), undec-10-yn-1-ol (1.04 g), copper(I) iodide (118 mg), triethylamine (939 mg), and bis(triphenylphosphine)palladium(II) chloride (217 mg) were added to N,N-dimethylformamide (10 mL). After purging with nitrogen, the reaction was carried out at 70 °C for 3 hours. After completion of the reaction, the mixture was extracted with ethyl acetate (200 mL × 3). The combined organic phases were washed with saturated brine (200 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: methanol / dichloromethane) to give 400 mg of the title compound.

[0366] b) Preparation of 1-(6-(11-hydroxyundecyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0367] 1-(6-(11-Hydroxyundec-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (200 mg) and palladium / carbon (15.5 mg) were added to methanol (2 mL), and the mixture was stirred overnight at 50 °C under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with methanol (10 mL × 3). The filtrate was concentrated under reduced pressure to give 150 mg of the title compound.

[0368] c) Preparation of 11-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)undecanal

[0369] 1-(6-(11-Hydroxyundecyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (150 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (308 mg) were added to dichloromethane (2 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 h. After the reaction, the pH was adjusted to 8 with saturated aqueous sodium bicarbonate, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phase was washed with saturated brine (10 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 90 mg of the title compound.

[0370] d) Preparation of 1-(6-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undecyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0371] 11-(3-(2,4-Dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)undecanal (100 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octan-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (114 mg) were dissolved in 1,2-dichloroethane (1 mL) and stirred at room temperature for 30 min. Then sodium triacetoxyborohydride (103 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 h under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 20.7 mg of the title compound.

[0372] 11H NMR (400 MHz, DMSO-d6) δ 10.81 (d, J = 19.8 Hz, 1H), 10.52 (s, 1H), 8.31 (s, 2H), 8.08 (d, J = 23.7 Hz, 1H), 7.53 (d, J = 8.4 Hz, 1H), 7.44–7.34 (m, 5H), 6.96 (dd, J = 8.5, 1.3 Hz, 1H), 4.58 (q, J = 9.6, 8.4 Hz, 2H), 4.51 (s, 2H), 4.19 (dd, J = 19.3, 16.1, 10.1 Hz, 2H), 3.95 (s, 3H), 3.91 (t, J = 6.7 Hz, 2H), 3.61 (d, J = 4.3 Hz, 2H), 3.56 (d, J = 3.7 Hz, 2H), 3.21–3.14 (m, 2H), 3.13–3.07 (m, 1H), 2.97 (d, J = 12.4 Hz, 1H), 2.81 (q, J = 6.9 Hz, 1H), 2.75 (s, 2H), 2.72–2.60 (m, 4H), 2.12–2.04 (m, 2H), 1.64 (m, 4H), 1.40–1.24 (m, 4H), 1.24–1.19 (m, 9H), 0.92–0.85 (m, 3H).

[0373] LCMS m / z = 866 [M+1] +

[0374] Example 14 Preparation of 4-(11-(((S)-2-(4-chlorophenyl)-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0375]

[0376] a) Preparation of 11-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)undec-10-ynal

[0377] 2-(2,6-Dioxopiperidin-3-yl)-4-(11-hydroxyundec-1-yn-1-yl)isoindoline-1,3-dione (100 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (210 mg) were added to dichloromethane (2 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 h. After the reaction, dichloromethane (30 mL) was added. The organic phase was washed with saturated brine (50 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 40 mg of the title compound.

[0378] b) Preparation of 4-(11-(((S)-2-(4-chlorophenyl)-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0379] 11-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)undec-10-ynal (30 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octan-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (40 mg) were dissolved in 1,2-dichloroethane (0.5 mL) and stirred at room temperature for 30 min. Then sodium triacetoxyborohydride (30 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 h under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 37.1 mg of the title compound.

[0380] 11H NMR (400 MHz, DMSO-d6) δ 10.81 (d, J = 18.3 Hz, 1H), 8.25 (s, 1H), 8.08 (d, J = 25.7 Hz, 1H), 7.85 (dt, J = 6.8, 3.4 Hz, 1H), 7.83–7.76 (m, 2H), 7.38 (d, J = 12.8 Hz, 4H), 5.13 (dd, J = 12.8, 5.4 Hz, 1H), 4.60 (d, J = 14.1 Hz, 2H), 4.50 (t, J = 8.2 Hz, 2H), 4.28 - 4.10 (m, 2H), 3.61 (d, J = 4.7 Hz, 1H), 3.56 (d, J = 3.8 Hz, 1H), 3.14 (dd, J = 21.1, 11.0 Hz, 4H), 2.97 (d, J = 12.4 Hz, 1H), 2.94–2.87 (m, 1H), 2.83 (dd, J = 15.3, 6.8 Hz, 2H), 2.66 (d, J = 11.6 Hz, 2H), 2.12–1.99 (m, 2H), 1.96–1.85 (m, 1H), 1.73 (d, J = 7.8 Hz, 2H), 1.69 (dd, J = 27.8, 8.2 Hz, 2H), 1.60–1.53 (m, 2H), 1.49–1.40 (m, 2H), 1.34 (s, 2H), 1.22 (d, J = 13.7 Hz, 8H), 0.88 (dt, J = 28.2, 7.1 Hz, 3H).

[0381] LCMS m / z = 876 [M+1] +

[0382] Example 15 Preparation of 3-(5-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0383]

[0384] a) Preparation of 11-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)undec-10-ynal 3-(5-(11-Hydroxyundec-1-yn-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (100 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (199 mg) were added to dichloromethane (2 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 hours. After the reaction, dichloromethane (30 mL) was added, and the organic phase was washed with saturated brine (50 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to obtain 50 mg of the title compound.

[0385] b) Preparation of 3-(5-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0386] 11-(1-(2,6-Dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)undec-10-ynal (50 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octane-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (83 mg) were dissolved in 1,2-dichloroethane (1 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (50 mg) was added to the mixture, and the mixture was stirred at room temperature under a nitrogen atmosphere for 1 hour. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 6.2 mg of the title compound.

[0387] 11H NMR (400 MHz, DMSO-d6) δ 10.82 (d, J = 21.1 Hz, 2H), 8.11 - 8.05 (m, 1H), 7.45–7.28 (m, 4H), 7.22 (d, J = 1.2 Hz, 1H), 7.14–6.97 (m, 2H), 5.37 (dd, J = 12.8, 5.3 Hz, 1H), 4.71–4.00 (m, 5H), 3.61 (d, J = 5.1 Hz, 1H), 3.56 (d, J = 4.0 Hz, 1H), 3.25 (s, 3H), 3.16 (t, J = 11.5 Hz, 1H), 3.09 (d, J = 8.3 Hz, 1H), 3.00–2.87 (m, 1H), 2.81 (q, J = 7.6, 7.0 Hz, 2H), 2.73–2.57 (m, 5H), 2.39 (q, J = 7.3 Hz, 2H), 2.13–1.97 (m, 2H), 1.73 (d, J = 8.0 Hz, 1H), 1.65 (t, J = 8.8 Hz, 1H), 1.52 (dp, J = 13.9, 6.9 Hz, 2H), 1.34 (s, 4H), 1.22 (d, J = 16.4 Hz, 10H), 0.93 - 0.83 (m, 3H).

[0388] LCMS m / z = 877 [M+1] +

[0389] Example 16 Preparation of 1-(6-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0390]

[0391] a) Preparation of 11-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)undec-10-ynal

[0392] 1-(6-(11-Hydroxyundec-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (200 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (413 mg) were added to dichloromethane (2 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 h. After the reaction, dichloromethane (30 mL) was added. The organic phase was washed with saturated brine (50 mL×3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 40 mg of the title compound.

[0393] b) Preparation of 1-(6-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0394] 11-(3-(2,4-Dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)undec-10-ynal (40 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octane-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (46 mg) were dissolved in 1,2-dichloroethane (1 mL) and stirred at room temperature for 30 min. Then sodium triacetoxyborohydride (41.5 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 h under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 9 mg of the title compound.

[0395] 11H NMR (400 MHz, DMSO-d6) δ 10.83 (d, J = 21.5 Hz, 1H), 10.57 (s, 1H), 8.21 (s, 1H), 8.09 (d, J = 27.5 Hz, 1H), 7.69 (s, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.42–7.34 (m, 4H), 7.06 (dd, J = 8.5, 1.3 Hz, 1H), 4.58 (s, 2H), 4.50 (s, 2H), 4.19 (m, 1H), 3.97 (s, 3H), 3.92 (t, J = 6.6 Hz, 2H), 3.61 (d, J = 5.1 Hz, 1H), 3.55 (d, J = 4.1 Hz, 1H), 3.32 (s, 3H), 3.16 (t, J = 10.6 Hz, 1H), 2.97 (d, J = 12.5 Hz, 1H), 2.81 (d, J = 7.0 Hz, 2H), 2.75 (t, J = 6.6 Hz, 3H), 2.67 (dt, J = 3.7, 1.9 Hz, 3H), 2.33 (p, J = 1.8 Hz, 1H), 2.11–2.01 (m, 1H), 1.72 (s, 1H), 1.66 (d, J = 8.1 Hz, 1H), 1.54 (dd, J = 14.0, 7.3 Hz, 2H), 1.34 (s, 4H), 1.23 (d, J = 15.6 Hz, 7H), 0.88 (m, 3H).

[0396] LCMS m / z = 862 [M+1] +

[0397] Example 17 Preparation of 3-(6-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undecyl)-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione

[0398]

[0399] a) Preparation of 3-(6-bromo-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione

[0400] 6-Bromo-2H-phthalazin-1-one (1 g), 3-bromopiperidine-2,6-dione (1.02 g), cesium carbonate (2.9 g), and tetrahydrofuran (20 mL) were added to a reaction flask, and the mixture was stirred under a nitrogen atmosphere at 100 °C for 15 hours. After the reaction was completed, the solid was filtered off, and the filter cake was washed with ethyl acetate (80 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was separated by reverse-phase column chromatography (column: C18, mobile phase: acetonitrile / water (0.1% NH3.H2O)) to obtain 750 mg of the title compound.

[0401] b) Preparation of 3-(6-(11-hydroxyundec-1-yn-1-yl)-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione

[0402] 3-(6-Bromo-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (750 mg), undec-10-yn-1-ol (563 mg), copper(I) iodide (85 mg), triethylamine (677 mg), and bis(triphenylphosphine)palladium(II) chloride (157 mg) were added to N,N-dimethylformamide (10 mL). After purging with nitrogen, the reaction was carried out at 70 °C for 3 hours. After the reaction was completed, the mixture was extracted with ethyl acetate (70 mL × 3), the organic phases were combined and washed with saturated brine (70 mL × 3), and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: methanol / dichloromethane = 0% - 10%) to obtain 360 mg of the title compound.

[0403] c) Preparation of 3-(6-(11-hydroxyundecyl)-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione

[0404] 3-(6-(11-Hydroxyundec-1-yn-1-yl)-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (150 mg) and palladium / carbon (45 mg) were added to methanol (5 mL), and the mixture was stirred at 50 °C under a hydrogen atmosphere for 15 hours. The resulting mixture was filtered, and the filter cake was washed with methanol (10 mL × 3). The filtrate was concentrated under reduced pressure to obtain 90 mg of the title compound.

[0405] d) Preparation of 11-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)undecanal

[0406] 3-(6-(11-Hydroxyundecyl)-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (90 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (178.5 mg) were added to dichloromethane (2 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 h. After the reaction, the pH was adjusted to 8 with saturated aqueous sodium bicarbonate, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phase was washed with saturated brine (10 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 80 mg of the title compound.

[0407] e) Preparation of 3-(6-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undecyl)-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione

[0408] 11-(2-(2,6-Dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)undecanal (80 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octan-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (132.5 mg) were dissolved in 1,2-dichloroethane (2 mL) and stirred at room temperature for 30 min. Then sodium triacetoxyborohydride (119.5 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 h under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 26.5 mg of the title compound.

[0409] 1H NMR (400 MHz, DMSO-d6) δ 11.03 (s, 1H), 10.83 (m, 1H), 8.42 (s, 1H), 8.25 (s, 1H), 8.18 (d, J = 8.1 Hz, 1H), 8.08 (m, 1H), 7.77 (s, 1H), 7.76 - 7.71 (m, 1H), 7.43 - 7.38 (m, 2H), 7.36 (m, 2H), 5.80 (dd, J = 12.4, 5.4 Hz, 1H), 4.64 - 4.54 (m, 2H), 4.54 - 4.43 (m, 1H), 4.32–4.02 (m, 2H), 3.58 (m, 2H), 3.21–3.06 (m, 2H), 3.00 - 2.87 (m, 2H), 2.84 - 2.74 (m, 3H), 2.73–2.62 (m, 3H), 2.62–2.54 (m, 2H), 2.19–2.04 (m, 1H), 1.96 - 1.85 (m, 1H), 1.80–1.53 (m, 4H), 1.41 - 1.09 (m, 18H), 0.88 (m, 3H).

[0410] LCMS m / z = 879 [M+1]+

[0411] Example 18 Preparation of 3-(6-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione

[0412]

[0413] a) Preparation of 11-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)undec-10-ynal

[0414] 3-(6-(11-Hydroxyundec-1-yn-1-yl)-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (100 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (200 mg) were added to dichloromethane (2 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 hours. After the reaction, the pH was adjusted to 8 with saturated aqueous sodium bicarbonate, and the mixture was extracted with dichloromethane (30 mL × 3). The organic phase was washed with saturated brine (15 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 85 mg of the title compound.

[0415] b) Preparation of 3-(6-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-3-oxopropyl)amino)undec-1-yn-1-yl)-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione

[0416] 11-(2-(2,6-Dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)undec-10-ynal (80 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octan-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (132.5 mg) were dissolved in 1,2-dichloroethane (2 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (119.5 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 10.1 mg of the title compound.

[0417] 11H NMR (400 MHz, DMSO-d6) δ 11.06 (s, 1H), 10.83 (d, J = 21.6 Hz, 1H), 8.44 (s, 1H), 8.21 (d, J = 8.3 Hz, 1H), 8.09 (m, 1H), 8.00 (s, 1H), 7.82 (d, J = 8.4 Hz, 1H), 7.38 (d, J = 12.8 Hz, 4H), 5.80 (dd, J = 12.7, 5.3 Hz, 1H), 4.71–4.39 (m, 4H), 4.30–4.09 (m, 2H), 3.58 (dd, J = 22.2, 4.6 Hz, 2H), 3.22 - 3.07 (m, 3H), 3.02–2.87 (m, 2H), 2.81 (d, J = 7.3 Hz, 1H), 2.67 (t, J = 9.2 Hz, 2H), 2.20–2.01 (m, 2H), 1.90 (s, 1H), 1.80–1.50 (m, 6H), 1.49–1.10 (m, 14H), 0.88 (dt, J = 29.3, 7.1 Hz, 3H).

[0418] LCMS m / z = 875 [M+1] +

[0419] Preparation of Example 19 4-((1R,5S)-8-((S)-2-(4-chlorophenyl)-3-((11-(4-(6-oxo-1,6-dihydropyridazin-3-yl))-2-(trifluoromethyl)phenyl)undecyl)amino)propanoyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-5-ethyl-5,8-dihydropteridin-7(6H)-one

[0420]

[0421] a) Preparation of methyl 4-(4-bromo-3-(trifluoromethyl)phenyl)-4-oxobutanoate

[0422] To a solution of sodium cyanide (60 mg) in N,N-dimethylformamide (30 mL) was added 4-bromo-3-(trifluoromethyl)benzaldehyde (3 g), and the mixture was stirred for 30 minutes under a nitrogen atmosphere at 35 °C. Then methyl acrylate (1.22 g) was added in portions. The resulting mixture was used directly in the next step without further purification.

[0423] b) Preparation of 6-(4-bromo-3-(trifluoromethyl)phenyl)-4,5-dihydropyridazin-3(2H)-one

[0424] Hydrazine hydrate (1.48 g, 98%) was added to the mixture obtained in the previous step, and the mixture was stirred at 60 °C under a nitrogen atmosphere for 8 h. After the reaction was completed, it was quenched with sodium hypochlorite at room temperature. The solid was filtered off, and ethyl acetate (300 mL) was added to the organic phase. The combined organic layers were washed with brine (100 mL×3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to obtain 1.4 g of the title compound.

[0425] c) Preparation of 6-(4-bromo-3-(trifluoromethyl)phenyl)pyridazin-3(2H)-one

[0426] 6-(4-Bromo-3-(trifluoromethyl)phenyl)-4,5-dihydropyridazin-3(2H)-one (1.4 g), copper(II) chloride (1.17 g) and acetonitrile (14 mL) were added to a reaction flask and stirred at 80 °C for 1 h. After the reaction was completed, the precipitated solid was collected by filtration, and the filter cake was washed with acetonitrile (30 mL×3). After the filter cake was dried, 500 mg of the target product was obtained.

[0427] d) Preparation of 6-(4-(11-hydroxyundec-1-yn-1-yl)-3-(trifluoromethyl)phenyl)pyridazin-3(2H)-one

[0428] 6-(4-Bromo-3-(trifluoromethyl)phenyl)pyridazin-3(2H)-one (500 mg), undec-10-yn-1-ol (527 mg), copper(I) iodide (60 mg), triethylamine (476 mg), and bis(triphenylphosphine)palladium(II) chloride (110 mg) were added to N,N-dimethylformamide (5 mL). After purging with nitrogen, the reaction was carried out at 70 °C for 3 h. After the reaction was completed, it was extracted with ethyl acetate (70 mL×3). The combined organic phases were washed with saturated brine (70 mL×3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: methanol / dichloromethane) to obtain 570 mg of the title compound.

[0429] e) Preparation of 6-(4-(11-hydroxyundecyl)-3-(trifluoromethyl)phenyl)pyridazin-3(2H)-one

[0430] 6-(4-(11-Hydroxyundec-1-yn-1-yl)-3-(trifluoromethyl)phenyl)pyridazin-3(2H)-one (100 mg) and palladium on carbon (60 mg) were added to methanol (5 mL), and the mixture was stirred at 50 °C overnight under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with methanol (10 mL×3). The filtrate was concentrated under reduced pressure to obtain 50 mg of the title compound.

[0431] f) Preparation of 11-(4-(6-oxo-1,6-dihydropyridazin-3-yl)-2-(trifluoromethyl)phenyl)undecanal

[0432] 6-(4-(11-Hydroxyundecyl)-3-(trifluoromethyl)phenyl)pyridazin-3(2H)-one (50 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (103 mg) were added to dichloromethane (1 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 hours. After the reaction, the pH was adjusted to 8 with saturated aqueous sodium bicarbonate, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phase was washed with saturated brine (10 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 20 mg of the title compound.

[0433] g) Preparation of 4-((1R,5S)-8-((S)-2-(4-chlorophenyl)-3-((11-(4-(6-oxo-1,6-dihydropyridazin-3-yl))-2-(trifluoromethyl)phenyl)undecyl)amino)propanoyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-5-ethyl-5,8-dihydropteridin-7(6H)-one

[0434] 11-(4-(6-Oxo-1,6-dihydropyridazin-3-yl)-2-(trifluoromethyl)phenyl)undecanal (20 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octan-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (23 mg) were dissolved in 1,2-dichloroethane (1 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (21 mg) was added to the mixture, and the mixture was stirred at room temperature under a nitrogen atmosphere for 1 hour. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 4.2 mg of the title compound. 11H NMR (400 MHz, DMSO-d6) δ 10.84 (d, J = 18.5 Hz, 1H), 8.17–8.03 (m, 4H), 7.60 (d, J = 8.0 Hz, 1H), 7.38 (d, J = 12.8 Hz, 4H), 7.02 (d, J = 9.9 Hz, 1H), 4.59 (s, 2H), 4.51 (s, 2H), 4.21 (s, 1H), 4.15 (s, 1H). 3.61 (d, J = 4.5 Hz, 2H), 3.56 (d, J = 3.7 Hz, 2H), 3.17 (d, J = 12.4 Hz, 2H), 2.97 (d, J = 12.5 Hz, 1H), 2.81 (d, J = 7.2 Hz, 1H), 2.76 (s, 2H), 2.67 (dt, J = 3.8, 1.8 Hz, 1H), 2.33 (q, J = 1.8 Hz, 1H), 2.06 (d, J = 12.5 Hz, 1H), 1.91 (s, 1H), 1.82–1.63 (m, 2H), 1.58 (s, 2H), 1.33 (s, 4H), 1.25–1.14 (m, 12H), 0.89 (m, 3H).

[0435] LCMS m / z = 862 [M+1] +

[0436] Example 20 Preparation of 4-((1R,5S)-8-((S)-2-(4-chlorophenyl)-3-((11-(4-(6-oxo-1,6-dihydropyridazin-3-yl))-2-(trifluoromethyl)phenyl)undec-10-yn-1-yl)amino)propanoyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-5-ethyl-5,8-dihydropteridin-7(6H)-one

[0437] a) Preparation of 11-(4-(6-oxo-1,6-dihydropyridazin-3-yl)-2-(trifluoromethyl)phenyl)undec-10-ynal

[0438] 6-(4-(11-Hydroxyundec-1-yn-1-yl)-3-(trifluoromethyl)phenyl)pyridazin-3(2H)-one (100 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (209 mg) were added to dichloromethane (1 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 h. After the reaction, the pH was adjusted to 8 with saturated aqueous sodium bicarbonate, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phase was washed with saturated brine (10 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 40 mg of the title compound.

[0439] b) Preparation of 4-((1R,5S)-8-((S)-2-(4-chlorophenyl)-3-((11-(4-(6-oxo-1,6-dihydropyridazin-3-yl))-2-(trifluoromethyl)phenyl)undec-10-yn-1-yl)amino)propanoyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-5-ethyl-5,8-dihydropteridin-7(6H)-one

[0440] 11-(4-(6-Oxo-1,6-dihydropyridazin-3-yl)-2-(trifluoromethyl)phenyl)undec-10-ynal (38 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octan-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (45 mg) were dissolved in 1,2-dichloroethane (1 mL) and stirred at room temperature for 30 min. Then sodium triacetoxyborohydride (40 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 h under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 15.8 mg of the title compound.

[0441] 11H NMR (400 MHz, DMSO-d6) δ 13.71–13.03 (s, 1H), 10.82 (d, J = 18.3 Hz, 1H), 8.16 (d, J = 10.3 Hz, 2H), 8.08 (m, 2H), 7.72 (d, J = 8.1 Hz, 1H), 7.42–7.34 (m, 4H), 7.02 (d, J = 9.9 Hz, 1H), 4.58 (s, 2H), 4.50 (t, J = 10.6 Hz, 2H), 4.12 (m, 1H), 3.58 (m, 2H), 3.23–3.01 (m, 1H), 2.96 (d, J = 12.4 Hz, 1H), 2.81 (d, J = 7.1 Hz, 1H), 2.67 (t, J = 2.0 Hz, 1H), 2.37–2.25 (m, 2H), 2.08 (d, J = 12.6 Hz, 1H), 1.95–1.86 (m, 1H), 1.77–1.63 (m, 3H), 1.59–1.49 (m, 3H), 1.36 (m, 5H), 1.22 (m, 9H), 0.88 (m, 3H).

[0442] LCMS m / z = 858 [M+1] +

[0443] Example 21 3-(4-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydro-pteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)(methyl)amino)undecyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0444]

[0445] a) Preparation of (S)-3-((tert-butoxycarbonyl)(methyl)amino)-2-(4-chlorophenyl)propanoic acid

[0446] A solution of (2S)-3-[(tert-butoxycarbonyl)amino]-2-(4-chlorophenyl)propanoic acid (500 mg) in tetrahydrofuran (5 mL) was cooled to 0 °C, and then sodium hydride (100 mg) was added portionwise over 30 minutes under nitrogen protection. Keeping at 0 °C and under a nitrogen atmosphere, iodomethane (710 mg) was added portionwise. Then the mixture was stirred at room temperature for an additional 3 hours. After completion of the reaction, the temperature was lowered to 0 °C and the reaction was quenched with water. The resulting mixture was extracted with ethyl acetate (50 mL × 3), the combined organic layers were washed with saturated brine (10 mL), and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 400 mg of the title compound.

[0447] b) Preparation of tert-butyl ((S)-2-(4-chlorophenyl)-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)(methyl)carbamate

[0448] The hydrochloride of 4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-5-ethyl-5,8-dihydropteridin-7(6H)-one (200 mg), (S)-3-((tert-butoxycarbonyl)(methyl)amino)-2-(4-chlorophenyl)propanoic acid (232 mg), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (468 mg), N,N-diisopropylethylamine (796 mg), and N,N-dimethylformamide (2 mL) were added to a reaction flask, and the mixture was stirred at room temperature under a nitrogen atmosphere for 3 hours. After completion of the reaction, ethyl acetate (30 mL) was added, and the organic layer was washed with brine (10 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 100 mg of the title compound.

[0449] c) Preparation of 4-((1R,5S)-8-((S)-2-(4-chlorophenyl)-3-(methylamino)propanoyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-5-ethyl-5,8-dihydropteridin-7(6H)-one

[0450] tert-Butyl ((S)-2-(4-chlorophenyl)-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)(methyl)carbamate (100 mg) was added to a 1,4-dioxane hydrochloric acid solution (4 M, 5 mL), and the reaction mixture was further stirred at room temperature for 2 hours. After completion of the reaction, the resulting mixture was concentrated under reduced pressure to give 80 mg of the title compound.

[0451] d) Preparation of 3-(4-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)(methyl)amino)undecyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0452] 11-(1-(2,6-Dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)undecanal (50 mg) and 4-((1R,5S)-8-((S)-2-(4-chlorophenyl)-3-(methylamino)propanoyl)-3,8-diazabicyclo[3.2.1]octane-3-yl)-5-ethyl-5,8-dihydropteridin-7(6H)-one (45 mg) were dissolved in 1,2-dichloroethane (2 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (39.5 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 29.5 mg of the title compound.

[0453] 11H NMR (400 MHz, DMSO-d6) δ 11.08 (s, 1H), 10.82 (d, J = 18.6 Hz, 1H), 8.08 (d, J = 21.7 Hz, 1H), 7.52–7.20 (m, 4H), 6.95 (d, J = 4.8 Hz, 2H), 6.88–6.79 (m, 1H), 5.36 (dd, J = 12.5, 5.4 Hz, 1H), 4.70 (d, J = 9.1 Hz, 1H), 4.52 (dd, J = 39.0, 15.5 Hz, 3H), 4.30 (d, J = 12.6 Hz, 1H), 4.18 (dd, J = 15.8, 9.9 Hz, 2H), 3.64–3.50 (m, 3H), 3.12 (m, 3H), 2.95 (d, J = 12.0 Hz, 2H), 2.86–2.79 (m, 1H), 2.71 (dd, J = 12.6, 8.1 Hz, 2H), 2.62 (d, J = 18.3 Hz, 2H), 2.42 (s, 1H), 2.35 (s, 1H), 2.26 (d, J = 13.9 Hz, 2H), 2.17 (d, J = 12.9 Hz, 3H), 2.04–1.90 (m, 2H), 1.72 (q, J = 11.2, 10.2 Hz, 2H), 1.58 (d, J = 8.1 Hz, 2H), 1.31 (m, 8H), 1.15 (m, 8H), 0.89 (m, 3H).

[0454] LCMS m / z = 895 [M+1] +

[0455] Example 22 Preparation of 3-(4-(11-(((S)-2-(4-chlorophenyl)-3-((R)-3-methyl-4-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-3-oxopropyl)amino)undecyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0456]

[0457] a) Preparation of tert-butyl (R)-3-methyl-4-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)piperazine-1-carboxylate

[0458] (R)-4-Chloro-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (1 g), tert-butyl (R)-3-methylpiperazine-1-carboxylate (10 g), N,N-diisopropylethylamine (6.5 g), and acetonitrile (30 mL) were added to a reaction flask, and the mixture was stirred at 100 °C for 48 h under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to obtain 1.1 g of the title compound.

[0459] b) Preparation of (R)-5-methyl-4-((R)-2-methylpiperazin-1-yl)-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one

[0460] tert-Butyl (R)-3-methyl-4-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)piperazine-1-carboxylate (200 mg) was added to a 1,4-dioxane hydrochloride solution (4 M, 5 mL), and the reaction solution was stirred at room temperature for an additional 1 h. After completion of the reaction, the resulting mixture was concentrated under reduced pressure to obtain 120 mg of the title compound.

[0461] c) Preparation of tert-butyl ((S)-2-(4-chlorophenyl)-3-((R)-3-methyl-4-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-3-oxopropyl)carbamate

[0462] (S)-3-((tert-Butoxycarbonyl)amino)-2-(4-chlorophenyl)propanoic acid (138 mg), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (58 mg), N,N-diisopropylethylamine (20 mg), and N,N-dimethylformamide (2 mL) were added to a reaction flask. After the mixture was stirred at room temperature under a nitrogen atmosphere for 10 min, (R)-5-methyl-4-((R)-2-methylpiperazin-1-yl)-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (100 mg) was added, and stirring was continued at room temperature for 3 h. After completion of the reaction, ethyl acetate (30 mL) was added, and the organic layer was washed with brine (10 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to obtain 120 mg of the title compound.

[0463] d) Preparation of (R)-4-((R)-4-((S)-3-amino-2-(4-chlorophenyl)propanoyl)-2-methylpiperazin-1-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one

[0464] (R)-4-((R)-4-((S)-3-Amino-2-(4-chlorophenyl)propanoyl)-2-methylpiperazin-1-yl)-5-methyl-5,8-dihydropyridinyl[2,3-d]pyrimidin-7(6H)-one (120 mg) was added to a 1,4-dioxane hydrochloride solution (4 M, 5 mL), and the reaction solution was stirred at room temperature for an additional 2 hours. After completion of the reaction, the resulting mixture was concentrated under reduced pressure to give 80 mg of the title compound.

[0465] e) Preparation of 3-(4-(11-(((S)-2-(4-chlorophenyl)-3-((R)-3-methyl-4-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-3-oxopropyl)amino)undecyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0466] 11-(1-(2,6-Dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)undecanal (50 mg) and (R)-4-((R)-4-((S)-3-amino-2-(4-chlorophenyl)propanoyl)-2-methylpiperazin-1-yl)-5-methyl-5,8-dihydropyridinyl[2,3-d]pyrimidin-7(6H)-one (39 mg) were dissolved in 1,2-dichloroethane (2 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (72 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 5.7 mg of the title compound.

[0467] 11H NMR (400 MHz, DMSO-d6) δ 11.07 (s, 1H), 10.66 (d, J = 7.3 Hz, 1H), 8.41–8.23 (m, 1H), 7.48–7.23 (m, 4H), 7.04–6.89 (m, 2H), 6.88 - 6.82 (m, 1H), 5.35 (dd, J = 12.5, 5.4 Hz, 1H), 4.31 - 4.14 (m, 1H), 3.94 - 3.64 (m, 2H), 3.54 (s, 3H), 3.49 - 3.39 (m, 1H), 3.23 - 3.15 (m, 2H), 3.13 - 3.06 (m, 2H), 2.94–2.83 (m, 4H), 2.81–2.68 (m, 1H), 2.68–2.58 (m, 4H), 2.36 - 2.21 (m, 1H), 2.05–1.94 (m, 1H), 1.66 - 1.49 (m, 2H), 1.42–1.15 (m, 18H), 1.07 - 0.93 (m, 4H), 0.85 (t, J = 6.7 Hz, 1H), 0.71 (dd, J = 63.4, 6.3 Hz, 2H).

[0468] LCMS m / z = 854 [M+1] +

[0469] Example 23 Preparation of 3-(4-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)amino)dodecyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0470]

[0471] a) Preparation of 2-(dec-9-yn-1-yl)-2-methyl-1,3-dioxolane

[0472] A toluene (10 mL) solution of dodec-11-yn-2-one (1.2 g) and ethylene glycol (825 mg) was stirred at 110 °C for 12 h. After completion of the reaction, the reaction was quenched with water (200 mL) and extracted with ethyl acetate (60 mL × 3). The combined organic phases were washed with brine (30 mL × 3) and dried over anhydrous sodium sulfate. The solid was filtered off and the filter cake was washed with ethyl acetate. The filtrate was concentrated under reduced pressure and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 600 mg of the title compound.

[0473] b) Preparation of 3-(3-Methyl-4-(10-(2-methyl-1,3-dioxolan-2-yl)dec-1-yn-1-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0474] 3-(4-Bromo-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (1.085 g), 2-(dec-9-yn-1-yl)-2-methyl-1,3-dioxolane (600 mg), cuprous iodide (102 mg), triethylamine (5 mL), and bis(triphenylphosphine)palladium(II) chloride (188 mg) were added to N,N-dimethylformamide (5 mL). After purging with nitrogen, the reaction was carried out at 70 °C for 3 h. After completion of the reaction, water (200 mL) was added to quench the reaction. The mixture was extracted with ethyl acetate (70 mL × 3), and the combined organic phases were washed with saturated brine (70 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to obtain 110 mg of the title compound.

[0475] c) Preparation of 3-(3-Methyl-4-(10-(2-methyl-1,3-dioxolan-2-yl)decyl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0476] 3-(3-Methyl-4-(10-(2-methyl-1,3-dioxolan-2-yl)dec-1-yn-1-yl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (100 mg) and palladium / carbon (22 mg) were added to methanol (10 mL). The mixture was stirred at 50 °C under a hydrogen atmosphere for 15 h. The resulting mixture was filtered, and the filter cake was washed with methanol (10 mL × 3). The filtrate was concentrated under reduced pressure to obtain 95 mg of the title compound.

[0477] d) Preparation of 3-(3-Methyl-2-oxo-4-(11-oxododecyl)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0478] 3-(3-Methyl-4-(10-(2-methyl-1,3-dioxolan-2-yl)decyl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (95 mg) was added to 1,4-dioxane hydrochloride solution (4 M, 1 mL), and the reaction mixture was stirred at room temperature for an additional 2 h. After completion of the reaction, the resulting mixture was concentrated under reduced pressure to obtain 77 mg of the title compound.

[0479] e) Preparation of 3-(4-(11-(((S)-2-(4-chlorophenyl))-3-((1R,5S)-3-(5-ethyl-7-oxo-5,6,7,8-tetrahydropteridin-4-yl)-3,8-diazabicyclo[3.2.1]oct-8-yl)-3-oxopropyl)amino)dodecyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione 3-(3-Methyl-2-oxo-4-(11-oxododecyl)-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (40 mg) and 4-{8-[(2S)-3-amino-2-(4-chlorophenyl)propanoyl]-3,8-diazabicyclo[3.2.1]octane-3-yl}-5-ethyl-6,8-dihydropteridin-7-one (51 mg) were dissolved in 1,2-dichloroethane (2 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (58 mg) was added to the mixture, and the mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 4.2 mg of the title compound.

[0480] 1 H NMR (400 MHz, DMSO-d6) δ 11.09 (s, 1H), 10.83 (d, J = 20.2 Hz, 1H), 8.08 (d, J = 23.4 Hz, 1H), 7.38 (d, J = 13.3 Hz, 4H), 6.95 (d, J = 5.2 Hz, 2H), 6.89–6.75 (m, 1H), 5.36 (dd, J = 12.6, 5.5 Hz, 1H), 4.56 (d, J = 16.1 Hz, 1H), 4.49 (s, 2H), 4.20 (s, 1H), 4.09 (s, 1H), 3.61 (d, J = 4.3 Hz, 1H), 3.54 (s, 4H), 3.19 (dd, J = 9.8, 8.1 Hz, 4H), 2.95 (s, 4H), 2.87 (s, 1H), 2.80–2.72 (m, 2H), 2.70–2.64 (m, 1H), 2.60 (s, 1H), 2.07–1.91 (m, 2H), 1.73 (s, 2H), 1.57 (s, 3H), 1.42–1.29 (m, 3H), 1.35–1.07 (m, 13H), 0.98–0.87 (m, 3H), 0.84 (t, J = 7.1 Hz, 2H). LCMS m / z = 895 [M+1] +

[0481] Preparation of 1-(6-(7-(4-((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)piperazin-1-yl)heptyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0482]

[0483] a) Preparation of 1-(6-(7-hydroxyhept-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0484] 1-(6-Bromo-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (300 mg), hept-6-yn-1-ol (125 mg), copper(I) iodide (35.5 mg), triethylamine (471 mg), and bis(triphenylphosphine)palladium(II) chloride (65 mg) were added to N,N-dimethylformamide (9 mL). After purging with nitrogen, the reaction was carried out at 70 °C for 3 hours. After completion of the reaction, the mixture was extracted with ethyl acetate (200 mL × 3). The combined organic phases were washed with saturated brine (200 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 285 mg of the title compound.

[0485] b) Preparation of 1-(6-(7-hydroxyheptyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0486] 1-(6-(7-Hydroxyhept-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (285 mg) and palladium / carbon (284.5 mg) were added to ethyl acetate (5 mL). The mixture was stirred at 50 °C overnight under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with ethyl acetate (10 mL × 3). The filtrate was concentrated under reduced pressure to give 170 mg of the title compound.

[0487] c) Preparation of 7-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)heptanal

[0488] 1-(6-(7-Hydroxyhept-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (170 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (121 mg) were added to dichloromethane (2 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 h. After the reaction, the pH was adjusted to 8 with saturated aqueous sodium bicarbonate, and the mixture was extracted with dichloromethane (30 mL × 3). The organic phase was washed with saturated brine (10 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 100 mg of the title compound.

[0489] d) Preparation of 1-(6-(7-(4-((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)piperazin-1-yl)heptyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0490] 7-(3-(2,4-Dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)heptanal (80 mg), (R)-4-((1S,6R)-5-((S)-2-(4-chlorophenyl)-3-(piperazin-1-yl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (38 mg), titanium(IV) isopropoxide (106 mg), triethylamine (23 mg), and N,N-dimethylformamide (1 mL) were added to a reaction flask, and the mixture was stirred at 50 °C under a nitrogen atmosphere for 1 h. Then sodium triacetoxyborohydride (32 mg) was added to the mixture, and the mixture was stirred at room temperature under a nitrogen atmosphere for 1 h. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 11.3 mg of the title compound.

[0491] 11H NMR (400 MHz, DMSO-d6) δ 10.62–10.39 (m, 2H), 8.26–8.15 (m, 1H), 7.53 (d, J = 8.4 Hz, 1H), 7.45–7.38 (m, 1H), 7.36 (d, J = 9.5 Hz, 2H), 7.34–7.07 (m, 2H), 6.96 (dt, J = 8.5, 1.6 Hz, 1H), 4.49 (t, J = 7.1 Hz, 2H), 4.26–3.95 (m, 3H), 3.90 (t, J = 6.7 Hz, 2H), 3.82–3.69 (m, 1H), 3.59 (d, J = 11.9 Hz, 1H), 3.51 (s, 1H), 3.17 (d, J = 9.5 Hz, 2H), 3.02 (dd, J = 21.9, 13.9 Hz, 2H), 2.76 (q, J = 6.8 Hz, 3H), 2.70 (s, 2H), 2.45 (s, 4H), 2.32–2.03 (m, 6H), 2.01–1.94 (m, 1H), 1.62 (s, 2H), 1.29 (s, 5H), 1.24 (s, 5H), 1.08 (t, J = 7.7 Hz, 2H), 0.95 (d, J = 6.9 Hz, 2H).

[0492] LCMS m / z = 850 [M+1] +

[0493] Example 25 Preparation of 1-(6-(5-(4-((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)piperazin-1-yl)pentyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0494]

[0495] a) Preparation of 1-(6-(5-hydroxypent-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0496] 1-(6-Bromo-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (130 mg), pent-4-yn-1-ol (20 mg), copper(I) iodide (8 mg), triethylamine (102 mg), and bis(triphenylphosphine)palladium(II) chloride (14 mg) were added to N,N-dimethylformamide (2 mL). After purging with nitrogen, the reaction was carried out at 70 °C for 3 hours. After completion of the reaction, the mixture was extracted with ethyl acetate (200 mL × 3). The combined organic phases were washed with saturated brine (200 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 110 mg of the title compound.

[0497] b) Preparation of 1-(6-(5-Hydroxypentyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0498] 1-(6-(5-Hydroxypent-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (110 mg) and palladium / carbon (110 mg) were added to ethyl acetate (5 mL). The mixture was stirred overnight at 50 °C under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with ethyl acetate (10 mL × 3). The filtrate was concentrated under reduced pressure to give 100 mg of the title compound.

[0499] c) Preparation of 5-(3-(2,4-Dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)pentanal

[0500] 1-(6-(5-Hydroxypentyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (100 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (257 mg) were added to dichloromethane (2 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 hours. After the reaction, the pH was adjusted to 8 with saturated aqueous sodium bicarbonate, and the mixture was extracted with dichloromethane (30 mL × 3). The organic phase was washed with saturated brine (10 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 70 mg of the title compound.

[0501] d) Preparation of 1-(6-(5-(4-((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)piperazin-1-yl)pentyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0502] Dissolve 5-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)pentanal (51.5 mg) and (R)-4-((1S,6R)-5-((S)-2-(4-chlorophenyl)-3-(piperazin-1-yl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (80 mg) in 1,2-dichloroethane (1 mL) and stir at 50 °C for 30 minutes. Then add sodium triacetoxyborohydride (99 mg) to the mixture and stir at room temperature under a nitrogen atmosphere for 1 hour. After the reaction is completed, concentrate the resulting mixture under reduced pressure. The crude product is separated by preparative liquid chromatography to obtain 11.2 mg of the title compound.

[0503] 1 H NMR (400 MHz, DMSO-d6) δ 10.60–10.40 (m, 2H), 8.24–8.13 (m, 1H), 7.68–7.49 (m, 2H), 7.46–7.39 (m, 1H), 7.37 (d, J = 3.7 Hz, 1H), 7.35 (s, 2H), 7.00–6.92 (m, 1H), 4.49 (t, J = 7.0 Hz, 1H), 4.15–4.03 (m, 1H), 4.01–3.91 (m, 3H), 3.90–3.74 (m, 2H), 3.59 (d, J = 11.8 Hz, 1H), 3.50 (d, J = 6.2 Hz, 1H), 3.41 - 3.36 (m, 1H), 3.17 (d, J = 9.4 Hz, 3H), 3.09–2.90 (m, 2H), 2.75 (t, J = 6.7 Hz, 3H), 2.70 (d, J = 7.3 Hz, 2H), 2.56 - 2.5 (m, 5H), 2.27 (t, J = 15.8 Hz, 6H), 1.71–1.57 (m, 2H), 1.47 (d, J = 34.9 Hz, 2H), 1.33–1.20 (m, 3H), 1.08 (t, J = 8.1 Hz, 2H), 0.95 (d, J = 6.9 Hz, 2H).

[0504] LCMS m / z = 822 [M+1]+

[0505] Example 26 Preparation of 1-(6-(3-(4-((S)-2-(4-chlorophenyl)-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5),6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)piperazin-1-yl)propyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0506]

[0507] a) Preparation of tert-butyldiphenyl(prop-2-yn-1-yloxy)silane

[0508] Propargyl alcohol (2 g), imidazole (4.86 g), tert-butyldiphenylchlorosilane (14.7 g), and dichloromethane (20 mL) were added to a reaction flask, and the mixture was stirred for 12 hours under a nitrogen atmosphere at room temperature. After the reaction was completed, dichloromethane (180 mL) was added, and the organic layer was washed with brine (150 mL × 3) and then dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure to obtain the title compound (8 g).

[0509] b) Preparation of 1-(6-(3-((tert-butyldiphenylsilyl)oxy)prop-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0510] 1-(6-Bromo-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (500 mg), tert-butyldiphenyl(prop-2-yn-1-yloxy)silane (911 mg), copper(I) iodide (58 mg), 1,8-diazabicyclo(5.4.0)undec-7-ene (471 mg), and tetrakis(triphenylphosphine)palladium (178 mg) were added to tetrahydrofuran (5 mL). After purging with nitrogen, the reaction was carried out at 80 °C for 3 hours. After the reaction was completed, the solid was filtered off, and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (mobile phase: methanol / dichloromethane) to obtain 280 mg of the title compound.

[0511] c) Preparation of 1-(6-(3-hydroxyprop-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0512] 1-(6-(3-((tert-Butyldiphenylsilyl)oxy)prop-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (280 mg), tetrabutylammonium fluoride (272 mg), and tetrahydrofuran (3 mL) were added to a reaction flask, and the mixture was stirred under a nitrogen atmosphere at room temperature for 2 h. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was purified by column chromatography (mobile phase: methanol / dichloromethane) to give 120 mg of the title compound.

[0513] d) Preparation of 1-(6-(3-Hydroxypropyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0514] 1-(6-(3-Hydroxyprop-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (120 mg), palladium / carbon (60 mg), and palladium hydroxide (60 mg) were added to methanol (5 mL), and the mixture was stirred at 50 °C overnight under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with methanol (10 mL × 3). The filtrate was concentrated under reduced pressure to give 90 mg of the title compound.

[0515] e) Preparation of 3-(3-(2,4-Dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)propanal

[0516] 1-(6-(3-Hydroxypropyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (90 mg) and (1,1,1-Triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (252 mg) were added to dichloromethane (2 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 h. After the reaction, the pH was adjusted to 8 with saturated aqueous sodium bicarbonate, and the mixture was extracted with dichloromethane (30 mL × 3). The organic phase was washed with saturated brine (10 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 45 mg of the title compound.

[0517] f) Preparation of 1-(6-(3-(4-((S)-2-(4-Chlorophenyl)-3-((1R,6S)-5-((R)-5-Methyl-7-oxo-5),6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)piperazin-1-yl)propyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0518] Dissolve 3-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)propanal (17 mg) and (R)-4-((1S,6R)-5-((S)-2-(4-chlorophenyl)-3-(piperazin-1-yl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (30 mg, Intermediate 1) in 1,2-dichloroethane (1 mL) and stir at 50 °C for 30 minutes. Then add sodium triacetoxyborohydride (37 mg) to the mixture and stir at room temperature under a nitrogen atmosphere for 1 hour. After completion of the reaction, concentrate the resulting mixture under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 1.4 mg of the title compound.

[0519] LCMS m / z = 794 [M+1] +

[0520] Example 27 Preparation of 1-(6-(9-(4-((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)piperazin-1-yl)nonyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0521]

[0522] a) Preparation of 1-(6-(9-hydroxynon-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0523] Add 1-(6-bromo-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (150 mg), non-8-yn-1-ol (130 mg), copper(I) iodide (17 mg), triethylamine (140 mg), and bis(triphenylphosphine)palladium(II) chloride (32 mg) to N,N-dimethylformamide (2 mL). After purging with nitrogen, react at 70 °C for 3 hours. After completion of the reaction, extract with ethyl acetate (50 mL × 3), combine the organic phases, wash with saturated brine (50 mL × 3), and dry over anhydrous sodium sulfate. After filtering off the solid, concentrate the filtrate under reduced pressure and purify the crude product by column chromatography (mobile phase: methanol / dichloromethane) to obtain 130 mg of the title compound.

[0524] b) Preparation of 1-(6-(9-hydroxynonyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0525] 1-(6-(9-Hydroxynon-1-yn-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (130 mg) and palladium / carbon (40 mg) were added to ethyl acetate (5 mL), and the mixture was stirred at 50 °C overnight under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with ethyl acetate (10 mL × 3). The filtrate was concentrated under reduced pressure to obtain 98 mg of the title compound.

[0526] c) Preparation of 9-(3-(2,4-Dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)nonanal

[0527] 1-(6-(9-Hydroxynonyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (98 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (215 mg) were added to dichloromethane (2 mL). After purging with nitrogen, the mixture was stirred at room temperature for 2 hours. After the reaction, the pH was adjusted to 8 with saturated aqueous sodium bicarbonate, and the mixture was extracted with dichloromethane (30 mL × 3). The organic phase was washed with saturated brine (10 mL × 3) and dried over anhydrous sodium sulfate. After filtering off the solid, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to obtain 54 mg of the title compound.

[0528] d) Preparation of 1-(6-(9-(4-((S)-2-(4-Chlorophenyl))-3-((1R,6S)-5-((R)-5-Methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)piperazin-1-yl)nonyl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0529] 9-(3-(2,4-Dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)nonanal (30 mg) and (R)-4-((1S,6R)-5-((S)-2-(4-Chlorophenyl)-3-(piperazin-1-yl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (47 mg, Intermediate 1) were dissolved in 1,2-dichloroethane (1 mL) and stirred at room temperature for 30 minutes. Then sodium triacetoxyborohydride (49 mg) was added to the mixture, and the mixture was stirred at room temperature under a nitrogen atmosphere for 1 hour. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 3.2 mg of the title compound.

[0530] LCMS m / z = 878 [M+1] +

[0531] Example 28 Preparation of 1-(6-(4-(8-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)octyl)piperazin-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0532]

[0533] a) Preparation of tert-butyl 4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)piperazine-1-carboxylate

[0534] 1-(6-Bromo-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (1.5 g), tert-butyl 1-piperazinecarboxylate (2 g), N,N-diisopropylethylamine (3 g) and N-methylpyrrolidone (20 mL) were added to a reaction flask. After purging with nitrogen, the reaction was carried out at 90 °C for 16 hours. After completion of the reaction, filtration was performed by suction, the filtrate was concentrated, and the crude product was purified by column chromatography (mobile phase: methanol / dichloromethane) to obtain 500 mg of the title compound.

[0535] b) Preparation of 1-(1-methyl-6-(piperazin-1-yl)-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0536] tert-Butyl 4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)piperazine-1-carboxylate (303 mg) was dissolved in 5 mL of hydrochloric acid methanol solution and stirred at room temperature for 2 hours. After completion of the reaction, concentration under reduced pressure was carried out to obtain the title compound (260 mg).

[0537] c) Preparation of 1-(6-(4-(8-hydroxyoctyl)piperazin-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0538] 1-(1-Methyl-6-(piperazin-1-yl)-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (50 mg), 8-bromooctanol (28.5 mg), triethylamine (28 mg), and potassium iodide (25 mg) were dissolved in N,N-dimethylformamide (5 mL). After purging with nitrogen, the reaction was carried out at room temperature for 2 hours. After completion of the reaction, the mixture was extracted with ethyl acetate and water. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 50 mg of the title compound.

[0539] d) Preparation of 8-(4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)piperazin-1-yl)octanal

[0540] 1-(6-(4-(8-Hydroxyoctyl)piperazin-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (19 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (21 mg) were dissolved in dichloromethane (1 mL). After purging with nitrogen, the reaction was carried out at room temperature for 2 hours. After completion of the reaction, the product was separated by silica gel preparative plate (eluent: methanol / dichloromethane) to obtain 11 mg of the product.

[0541] e) Preparation of 1-(6-(4-(8-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)octyl)piperazin-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione

[0542] 8-(4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)piperazin-1-yl)octanal (11 mg) and (R)-4-((1S,6R)-5-((S)-3-amino-2-(4-chlorophenyl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (11 mg) were dissolved in N,N-dimethylformamide (1 mL). Sodium triacetoxyborohydride (8 mg) was added, and after purging with nitrogen, the reaction was carried out at room temperature for 2 hours. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 4.5 mg of the title compound.

[0543] 11H NMR (400 MHz, DMSO-d6) δ 10.49 (s, 1H), 10.46 (s, 1H), 8.36 (s, 1H), 8.20 (s, 1H), 7.46–7.43 (m, 1H), 7.38 (s, 1H), 7.35 (m, 2H), 6.91 (d, J = 8.9 Hz, 1H), 6.80 (d, J = 7.3 Hz, 1H), 4.45–4.42 (m, 1H), 4.11 (s, 2H), 3.89 (m, 5H), 3.20 (m, 6H), 3.15 (s, 4H), 2.76–2.69 (m, 3H), 2.67 (s, 1H), 2.35–2.20 (m, 4H), 2.03–1.99 (m, 1H), 1.45 (s, 3H), 1.39–1.13 (m, 14H), 1.10–1.06 (m, 1H), 0.96 (m, 3H), 0.87–0.84 (m, 1H).

[0544] LCMS m / z = 879 [M+1] +

[0545] Example 29 Preparation of 5-(4-(8-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)octyl)piperazin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0546]

[0547] a) Preparation of tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazine-1-carboxylate

[0548] 2-(2,6-Dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (2 g), tert-butyl piperazine-1-carboxylate (2.7 g), N,N-diisopropylethylamine (3.74 g) and N-methylpyrrolidone (25 mL) were added to a reaction flask. After purging with nitrogen, the reaction was carried out at 90 °C for 16 hours. After completion of the reaction, filtration was carried out by suction, the filtrate was concentrated, and the crude product was purified by column chromatography (mobile phase: methanol / dichloromethane) to obtain 1.3 g of the title compound.

[0549] b) Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindoline-1,3-dione

[0550] tert-Butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazine-1-carboxylate (1.3 g), trifluoroacetic acid (2.5 mL), and dichloromethane (5 mL) were added to a reaction flask and reacted at room temperature for 1 hour. After the reaction was completed, filtration was carried out by suction, the filtrate was concentrated, and the crude product was purified by column chromatography (mobile phase: methanol / dichloromethane) to obtain 900 mg of the title compound.

[0551] c) Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(4-(octan-8-yloxy)piperazin-1-yl)isoindoline-1,3-dione

[0552] 2-(2,6-Dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindoline-1,3-dione (150 mg), 8-bromooctan-1-ol (68 mg), potassium iodide (55 mg), and triethylamine (133 mg) were dissolved in N,N-dimethylformamide (4 mL) solution. After purging with nitrogen, the reaction was carried out at room temperature for 20 hours. After the reaction was completed, filtration was carried out by suction, the filtrate was concentrated, and the crude product was purified by column chromatography (mobile phase: methanol / dichloromethane) to obtain 118 mg of the title compound. d) Preparation of 8-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)octanal

[0553] 2-(2,6-Dioxopiperidin-3-yl)-5-(4-(octan-8-yloxy)piperazin-1-yl)isoindoline-1,3-dione (118 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (128 mg) were dissolved in dichloromethane (4 mL) solution. After purging with nitrogen, the reaction was carried out at room temperature for 2 hours. After the reaction was completed, filtration was carried out by suction, the filtrate was concentrated, and the crude product was purified by column chromatography (mobile phase: methanol / dichloromethane = 0% - 10%) to obtain 87 mg of the title compound.

[0554] e) Preparation of 5-(4-(8-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)octyl)piperazin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0555] 8-(4-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)octanal (20 mg), (R)-4-((1S,6R)-5-((S)-3-Amino-2-(4-chlorophenyl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (10 mg) were dissolved in N,N-dimethylformamide (3 mL) solution. Sodium triacetoxyborohydride (14 mg) was added. After purging with nitrogen, the reaction was carried out at room temperature for 2 hours. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 1.04 mg of the title compound.

[0556] 1 H NMR (400 MHz, DMSO-d6) δ 11.07 (s, 2H), 10.49 (d, J = 11.3 Hz, 1H), 8.19 (s, 1H), 7.67 (d, J = 8.5 Hz, 1H), 7.44–7.29 (m, 4H), 7.25 (dd, J = 8.7, 2.3 Hz, 1H), 5.07 (dd, J = 12.9, 5.4 Hz, 1H), 4.46 (t, J = 7.0 Hz, 1H), 4.11 (d, J = 9.3 Hz, 1H), 3.97–3.83 (m, 1H), 3.83–3.73 (m, 1H), 3.65 (d, J = 10.9 Hz, 1H), 3.56–3.38 (m, 6H), 3.23–3.05 (m, 4H), 2.95–2.64 (m, 4H), 2.64–2.52 (m, 2H), 2.48–2.37 (m, 4H), 2.34–2.20 (m, 3H), 2.01 (ddd, J = 8.0, 6.1, 2.2 Hz, 2H), 1.51–1.32 (m, 5H), 1.27–1.20 (m, 9H), 0.94 (d, J = 6.9 Hz, 2H).

[0557] LCMS m / z = 893 [M+1] +

[0558] Example 30 Preparation of 5-(4-(6-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)hexyl)piperazin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0559]

[0560] a) Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(4-(6-hydroxyhexyl)piperazin-1-yl)isoindoline-1,3-dione

[0561] Dissolve 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindoline-1,3-dione (150 mg), 6-bromohexan-1-ol (59 mg), potassium iodide (55 mg), and triethylamine (133 mg) in N,N-dimethylformamide (4 mL) solution. After purging with nitrogen, react at room temperature for 20 hours. After the reaction is completed, filter by suction, concentrate the filtrate, and purify the crude product by column chromatography (mobile phase: methanol / dichloromethane) to obtain 86 mg of the title compound.

[0562] b) Preparation of 6-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)hexanal

[0563] Dissolve 2-(2,6-dioxopiperidin-3-yl)-5-(4-(6-hydroxyhexyl)piperazin-1-yl)isoindoline-1,3-dione (86 mg) and (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (99 mg) in dichloromethane (4 mL) solution. After purging with nitrogen, react at room temperature for 2 hours. After the reaction is completed, filter by suction, concentrate the filtrate, and purify the crude product by column chromatography (mobile phase: methanol / dichloromethane) to obtain 57 mg of the title compound.

[0564] c) Preparation of 5-(4-(6-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)hexyl)piperazin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0565] 6-(4-(2-(2,6-Dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)hexanal (45 mg), (R)-4-((1S,6R)-5-((S)-3-Amino-2-(4-chlorophenyl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (15 mg) were dissolved in N,N-dimethylformamide (3 mL). Sodium triacetoxyborohydride (22 mg) was added. After purging with nitrogen, the reaction was carried out at room temperature for 2 hours. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 2.45 mg of the title compound.

[0566] 1 H NMR (400 MHz, DMSO-d6) δ 11.07 (s, 2H), 10.48 (d, J = 10.8 Hz, 1H), 8.20 (d, J = 8.4 Hz, 1H), 7.66 (t, J = 7.9 Hz, 1H), 7.36 (dd, J = 16.4, 11.8 Hz, 3H), 7.24 (d, J = 9.5 Hz, 1H), 5.07 (dd, J = 13.2, 5.4 Hz, 1H), 4.43 (d, J = 23.1 Hz, 1H), 4.12 (s, 1H), 4.05–3.8 (m, 3H), 3.75–3.49 (m, 10H), 3.25–3.03 (m, J = 12.4 Hz, 5H), 2.96–2.55 (m, 7H), 2.28 (q, J = 16.3, 14.9 Hz, 4H), 2.01 (d, J = 11.9 Hz, 1H), 1.47–1.13 (m, 7H), 1.08 (t, J = 8.4 Hz, 1H), 0.95 (t, J = 7.4 Hz, 2H).

[0567] LCMS m / z = 865 [M+1] +

[0568] Example 31 Preparation of 5-(4-(4-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)butyl)piperazin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0569]

[0570] a) Preparation of 4-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)butyraldehyde

[0571] Dissolve 2-(2,6-dioxopiperidin-3-yl)-5-(piperazin-1-yl)isoindoline-1,3-dione (236 mg), succinaldehyde (148 mg), sodium triacetoxyborohydride (147 mg), and triethylamine (93 mg) in acetonitrile (4 mL). After purging with nitrogen, react at room temperature for 2 hours. After the reaction is completed, filter by suction, concentrate the filtrate, and purify the crude product by column chromatography (mobile phase: methanol / dichloromethane) to obtain 116 mg of the title compound.

[0572] b) Preparation of 5-(4-(4-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)butyl)piperazin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0573] Dissolve 4-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)butyraldehyde (32 mg) and (R)-4-((1S,6R)-5-((S)-3-amino-2-(4-chlorophenyl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (11 mg) in N,N-dimethylformamide (3 mL). Add sodium triacetoxyborohydride (25 mg), purge with nitrogen, and react at room temperature for 2 hours. After the reaction is completed, concentrate the resulting mixture under reduced pressure. The crude product is separated by preparative liquid chromatography to obtain 6.64 mg of the title compound. LCMS m / z = 837 [M+1] +

[0574] Example 32 Preparation of 5-(4-(((S)-2-(4-chlorophenyl)-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]heptan-2-yl)-3-oxopropyl)amino)-[1,4'-bipiperidin]-1'-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0575]

[0576] a) Preparation of tert-butyl 4-(((S)-2-(4-chlorophenyl)-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)piperidine-1-carboxylate

[0577] (R)-4-((1S,6R)-5-((S)-3-amino-2-(4-chlorophenyl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (200 mg), tert-butyl 4-oxopiperidine-1-carboxylate (90 mg), and 1,2-dichloroethane (2 mL) were placed in a reaction flask, and the mixture was stirred at room temperature for 1 hour. Then, sodium triacetoxyborohydride (288 mg) was added to the above mixture, and the resulting mixture was stirred at room temperature for another 2 hours. After the reaction was completed, the mixture was concentrated and directly used for the next step.

[0578] b) Preparation of (R)-4-((1S,6R)-5-((S)-2-(4-chlorophenyl)-3-(piperidin-4-ylamino)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one

[0579] tert-Butyl 4-(((S)-2-(4-chlorophenyl)-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)piperidine-1-carboxylate (100 mg) was added to 1,4-dioxane hydrochloride solution (4 M, 2 mL), and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the mixture was concentrated and directly used for the next step.

[0580] c) Preparation of 5-(4-(((S)-2-(4-chlorophenyl)-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]heptan-2-yl)-3-oxopropyl)amino)-[1,4'-bipiperidin]-1'-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0581] (R)-4-((1S,6R)-5-((S)-2-(4-chlorophenyl)-3-(piperidin-4-ylamino)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (60 mg), 2-(2,6-dioxopiperidin-3-yl)-5-(4-oxopiperidin-1-yl)isoindoline-1,3-dione (40 mg), titanium(IV) isopropoxide (98 mg), and dichloromethane (1 mL) were added to a reaction flask, and the mixture was stirred for 1 h at room temperature under a nitrogen atmosphere. Then sodium triacetoxyborohydride (73 mg) was added to the mixture, and the mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 19.8 mg of the title compound.

[0582] 1 H NMR (400 MHz, DMSO-d6) δ 11.07 (s, 1H), 10.49 (d, J = 9.7 Hz, 1H), 8.30–8.12 (m, 2H), 7.65 (d, J = 8.5 Hz, 1H), 7.45–7.36 (m, 3H), 7.34–7.26 (m, 1H), 7.26–7.19 (m, 1H), 5.06 (dd, J = 12.9, 5.4 Hz, 1H), 4.39 (m, 1H), 4.20–4.10 (m, 4H), 4.07 (d, J = 14.3 Hz, 1H), 3.93–3.81 (m, 1H), 3.77 (t, J = 6.5 Hz, 1H), 3.64 (d, J = 12.6 Hz, 2H), 3.49 (s, 2H), 3.35 (s, 2H), 3.20–3.08 (m, 2H), 2.98–2.85 (m, 3H), 2.77 (dt, J = 16.5, 6.1 Hz, 4H), 2.58–2.52 (m, 1H), 2.35–2.20 (m, 1H), 2.11 (m, 2H), 2.01 (dd, J = 9.3, 4.1 Hz, 1H), 1.79 (s, 4H), 1.45 (dd, J = 15.3, 8.3 Hz, 2H), 1.22 (d, J = 13.4 Hz, 2H), 1.07 (dd, J = 10.2, 6.8 Hz, 1H), 1.00–0.88 (m, 2H).

[0583] LCMS m / z = 863 [M+1] +

[0584] Preparation of Example 33: 5-(4-((4-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0585]

[0586] a) Preparation of 4-piperidinecarbaldehyde

[0587] tert-Butyl 4-formylpiperidine-1-carboxylate (500 mg) was added to 1,4-dioxane hydrochloride solution (4 M, 5 mL), and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, the mixture was concentrated and used directly for the next step.

[0588] b) Preparation of 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carbaldehyde

[0589] 4-Piperidinecarbaldehyde (255 mg), 2-(2,4-dioxocyclohexyl)-5-fluoroisoindoline-1,3-dione (391 mg), N,N-diisopropylethylamine (1101 mg) and dimethyl sulfoxide (3 mL) were added to a reaction flask and stirred at 50 °C for 12 hours. After completion of the reaction, the mixture was concentrated, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to obtain 130 mg of the title compound.

[0590] c) Preparation of 5-(4-((4-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0591] (R)-4-((1S,6R)-5-((S)-2-(4-chlorophenyl)-3-(piperidin-4-ylamino)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (69 mg), 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carbaldehyde (30 mg), titanium(IV) isopropoxide (23 mg), and 1,2-dichloroethane (1 mL) were added to a reaction flask, and the mixture was stirred for 1 h under a nitrogen atmosphere at room temperature. Then sodium triacetoxyborohydride (52 mg) was added to the mixture, and the mixture was stirred at room temperature under a nitrogen atmosphere for 2 h. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 8.8 mg of the title compound.

[0592] 1 H NMR (400 MHz, DMSO-d6) δ 11.06 (s, 1H), 10.47 (d, J = 9.5 Hz, 1H), 8.24–8.18 (m, 1H), 7.64 (dd, J = 8.5, 1.4 Hz, 1H), 7.40 (d, J = 2.4 Hz, 4H), 7.37–7.25 (m, 1H), 7.21 (d, J = 9.3 Hz, 1H), 5.05 (dd, J = 12.9, 5.4 Hz, 1H), 4.36 (m, 1H), 4.22–3.98 (m, 1H), 3.93–3.73 (m, 2H), 3.51 (d, J = 6.0 Hz, 3H), 2.91 (m, 4H), 2.79–2.66 (m, 4H), 2.65–2.54 (m, 4H), 2.35–2.18 (m, 1H), 2.08 (d, J = 8.3 Hz, 4H), 2.02 (s, 2H), 1.82 (d, J = 56.7 Hz, 6H), 1.26–0.90 (m, 9H).

[0593] LCMS m / z = 877 [M+1] +

[0594] Example 34 Preparation of 5-(4-((4-((((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)methyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0595]

[0596] a) Preparation of 5-(4-((4-(dimethoxymethyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0597] 4-(Dimethoxymethyl)piperidine (215.5 mg), 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carbaldehyde (500 mg), titanium(IV) isopropoxide (1.15 g), and 1,2-dichloroethane (5 mL) were added to a reaction flask, and the mixture was stirred for half an hour under a nitrogen atmosphere at room temperature. Then sodium triacetoxyborohydride (861 mg) was added to the mixture, and the mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. After completion of the reaction, the mixture was concentrated, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to obtain 220 mg of the title compound.

[0598] b) Preparation of 1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidine-4-carbaldehyde

[0599] 5-(4-((4-(dimethoxymethyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (120 mg) was added to a 1,4-dioxane hydrochloride solution (4 M, 2 mL), and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, the mixture was concentrated and directly used for the next step.

[0600] c) Preparation of 5-(4-((4-((((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)methyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0601] (R)-4-((1S,6R)-5-((S)-3-Amino-2-(4-chlorophenyl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (24 mg), 1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidine-4-carbaldehyde (25 mg), and tetrahydrofuran (1 mL) were added to a reaction flask, and the mixture was stirred for 1 h under a nitrogen atmosphere at room temperature. Then sodium triacetoxyborohydride (159.5 mg) was added to the mixture, and the mixture was stirred at room temperature under a nitrogen atmosphere for 2 h. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 16.2 mg of the title compound.

[0602] 1 H NMR (400 MHz, DMSO-d6) δ 11.03 (s, 1H), 10.48 (d, J = 10.2 Hz, 1H), 8.42 (s, 1H), 7.64 (d, J = 8.5 Hz, 1H), 7.39 (d, J = 4.6 Hz, 2H), 7.35 (d, J = 4.4 Hz, 2H), 7.29 (s, 1H), 7.21 (d, J = 8.7 Hz, 1H), 5.06 (dd, J = 12.8, 5.5 Hz, 1H), 4.42 (t, J = 10.0 Hz, 1H), 4.20–4.06 (m, 1H), 4.06–3.96 (m, 2H), 3.86–3.73 (m, 1H), 3.64 (d, J = 7.4 Hz, 1H), 3.51 (d, J = 6.6 Hz, 2H), 3.21–3.05 (m, 3H), 2.96 (d, J = 12.3 Hz, 3H), 2.90 (d, J = 13.5 Hz, 4H), 2.83–2.76 (m, 2H), 2.74 (d, J = 8.1 Hz, 3H), 2.70–2.57 (m, 3H), 2.43–2.34 (m, 2H), 2.27 (t, J = 16.4 Hz, 1H), 2.14–1.94 (m, 1H), 1.87–1.65 (m, 3H), 1.59 (d, J = 16.7 Hz, 2H), 1.28 (s, 1H), 1.07 (d, J = 13.7 Hz, 3H), 0.96 (dd, J = 15.3, 6.7 Hz, 3H).

[0603] LCMS m / z = 891 [M+1] +

[0604] Preparation of Example 35 5-(2-(4-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)piperidin-1-yl)-7-azaspiro[3.5]nonan-7-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0605]

[0606] a) Preparation of 7-azaspiro[3.5]nonan-2-one

[0607] tert-Butyl 2-oxo-7-azaspiro[3.5]nonane-7-carboxylate (1 g) was added to a 1,4-dioxane hydrochloric acid solution (4 M, 10 mL), and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, the mixture was concentrated and used directly in the next step.

[0608] b) Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(2-oxo-7-azaspiro[3.5]nonan-7-yl)isoindoline-1,3-dione 7-Azaspiro[3.5]nonan-2-one (300 mg), 2-(2,4-dioxocyclohexyl)-5-fluoroisoindoline-1,3-dione (472 mg), N,N-diisopropylethylamine (1324 mg) and dimethyl sulfoxide (5 mL) were added to a reaction flask and stirred at 100 °C for 12 hours. After completion of the reaction, the mixture was concentrated, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to obtain 250 mg of the title compound.

[0609] c) Preparation of 5-(2-(4-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)piperidin-1-yl)-7-azaspiro[3.5]nonan-7-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (R)-4-((1S,6R)-5-((S)-2-(4-chlorophenyl)-3-(piperidin-4-ylamino)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (93 mg), 2-(2,6-dioxopiperidin-3-yl)-5-(2-oxo-7-azaspiro[3.5]nonan-7-yl)isoindoline-1,3-dione (70 mg), and 1,2-dichloroethane (7 mL) were added to a reaction flask, and the mixture was stirred for 1 hour at room temperature under a nitrogen atmosphere. Then sodium triacetoxyborohydride (112.5 mg) was added to the mixture, and the mixture was stirred at room temperature for 2 hours under a nitrogen atmosphere. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 7.1 mg of the title compound.

[0610] 1 H NMR(400MHz,DMSO-d6)δ11.07(s,1H),10.50(s,1H),8.20(s,1H),7.65(d,J=8.4Hz,1H),7.44(s,2H),7.34(d,J=8.2Hz,3H),7.25(s,1H),5.06(d,J=12.0Hz,1H),4.75–4.43(m,1H),4.21–4.04(m,1H),3.87–3.66(m,3H),3.52–3.43(m,5H),3.20(s,5H),2.88(s,2H),2.76(d,J=14.6Hz,2H),2.67(s,1H),2.60(s,1H),2.36–2.15(m,1H),2.17–2.08(m,1H),2.09–1.94(m,1H),1.82–1.71(m,6H),1.69–1.48(m,2H),1.35–1.18(m,5H),1.07(d,J=7.3Hz,1H),0.99–0.85(m,1H),0.87–0.76(m,4H).

[0611] LCMS m / z=903[M+1] +

[0612] Preparation of Example 36: 5-(4-(2-(4-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)piperidin-1-yl)ethyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0613]

[0614] a) Preparation of 2-(2,6-dioxopiperidin-3-yl)-5-(4-(2-hydroxyethyl)piperidin-1-yl)isoindoline-1,3-dione

[0615] 2-(Piperidin-4-yl)ethan-1-ol (300 mg), 2-(2,4-dioxocyclohexyl)-5-fluoroisoindoline-1,3-dione (641 mg), N,N-diisopropylethylamine (1.8 g) and N-methylpyrrolidone (3 mL) were added to a reaction flask and stirred at 90 °C for 16 h. After completion of the reaction, water (10 mL) was added and the mixture was extracted with ethyl acetate (15 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate. The solid was filtered off, the organic phase was concentrated, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 450 mg of the title compound.

[0616] b) Preparation of 2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)acetaldehyde

[0617] 2-(2,6-Dioxopiperidin-3-yl)-5-(4-(2-hydroxyethyl)piperidin-1-yl)isoindoline-1,3-dione (450 mg), (1,1,1-triacetoxy)-1,1-dihydro-1,2-benziodoxol-3(1H)-one (2.47 g), and dichloromethane (10 mL) were added to a reaction flask and stirred at room temperature for 1 h. After completion of the reaction, water (15 mL) was added and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate. The solid was filtered off, the organic phase was concentrated, and the crude product was purified by column chromatography (mobile phase: ethyl acetate / petroleum ether) to give 350 mg of the title compound.

[0618] c) Preparation of 5-(4-(2-(4-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)piperidin-1-yl)ethyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0619] (R)-4-((1S,6R)-5-((S)-2-(4-chlorophenyl)-3-(piperidin-4-ylamino)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (68 mg), 2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)acetaldehyde (50 mg), titanium(IV) isopropoxide (56 mg), and tetrahydrofuran (2 mL) were added to a reaction flask, and the mixture was stirred for half an hour under a nitrogen atmosphere at room temperature. Then sodium triacetoxyborohydride (55 mg) was added to the mixture, and the mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 20.8 mg of the title compound.

[0620] 11H NMR (400 MHz, DMSO-d6) δ 11.01 (s, 1H), 10.43 (d, J = 8.0 Hz, 1H), 8.13–8.10 (m, 1H), 7.58 (d, J = 8.5 Hz, 1H), 7.31 (m, 4H), 7.23 (d, J = 2.3 Hz, 1H), 7.15 (dd, J = 8.7, 2.2 Hz, 1H), 4.99 (dd, J = 12.9, 5.4 Hz, 1H), 4.41–4.30 (m, 1H), 4.41–4.24 (m, 1H), 4.09–4.01 (m, 1H), 4.00–3.96 (m, 1H), 3.96–3.93 (m, 1H), 3.76–3.70 (m, 1H), 3.70–3.66 (m, 1H), 3.63–3.58 (m, 1H), 3.21–3.17 (m, 1H), 3.16–3.13 (m, 1H), 3.12–3.09 (m, 1H), 3.09–3.03 (m, 1H), 2.92–2.85 (m, 2H), 2.84–2.79 (m, 1H), 2.79–2.66 (m, 4H), 2.58–2.48 (m, 2H), 2.33–2.21 (m, 2H), 2.18 (d, J = 15.9 Hz, 2H), 1.96 (dd, J = 6.7, 3.9 Hz, 1H), 1.92 (d, J = 3.8 Hz, 3H), 1.75–1.61 (m, 4H), 1.58–1.42 (m, 1H), 1.31 (q, J = 7.7 Hz, 2H), 1.21–1.08 (m, 2H), 1.03–0.90 (m, 1H), 0.88 (t, J = 6.5 Hz, 3H), 0.09 (q, J = 5.2 Hz, 1H).

[0621] LCMS m / z = 892 [M+1] +

[0622] Example 37 Preparation of 5-(4-(2-(4-(2-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)ethyl)piperidin-1-yl)ethyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0623]

[0624] a) Preparation of 5-(4-(2-(4-(2,2-dimethoxyethyl)piperidin-1-yl)ethyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0625] A solution of 2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)acetaldehyde (100 mg) and 4-(2,2-dimethoxyethyl)piperidine (45 mg) in 1,2-dichloroethane (2 mL) was stirred at room temperature for half an hour. Then sodium triacetoxyborohydride (110.5 mg) was added to the above mixture and stirring was continued at room temperature for 1.5 hours. After the reaction was completed, the resulting mixture was diluted with water (5 mL) and extracted with ethyl acetate (5 mL × 3). The combined organic layers were washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and used directly for the next step.

[0626] b) Preparation of 2-(1-(2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)ethyl)piperidin-4-yl)acetaldehyde

[0627] 5-(4-(2-(4-(2,2-dimethoxyethyl)piperidin-1-yl)ethyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (90 mg) was added to a 1,4-dioxane hydrochloride solution (4 M, 1 mL) and stirred at room temperature for 1 hour. After the reaction was completed, the pH was adjusted to 8 - 9 with an aqueous sodium bicarbonate solution, and then extracted with ethyl acetate (10 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate. The solid was filtered off, and after concentration of the organic phase, the crude product was separated by preparative liquid chromatography to obtain 60 mg of the title compound.

[0628] c) Preparation of 5-(4-(2-(4-(2-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)ethyl)piperidin-1-yl)ethyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0629] (R)-4-((1S,6R)-5-((S)-3-Amino-2-(4-chlorophenyl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (44.5 mg), 2-(1-(2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)ethyl)piperidin-4-yl)acetaldehyde (50 mg), and 1,2-dichloroethane (2 mL) were added to a reaction flask, and the mixture was stirred for half an hour under a nitrogen atmosphere at room temperature. Then sodium triacetoxyborohydride (43 mg) was added to the mixture, and the mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. After the reaction was completed, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain 2.5 mg of the title compound.

[0630] 1 H NMR (400 MHz, DMSO-d6) δ 11.08 (s, 1H), 10.50 (d, J = 9.0 Hz, 1H), 8.22–8.18 (m, 1H), 7.64 (d, J = 8.6 Hz, 1H), 7.43–7.37 (m, 2H), 7.37–7.32 (m, 2H), 7.29 (d, J = 2.3 Hz, 1H), 7.22 (dd, J = 8.8, 2.3 Hz, 1H), 5.06 (dd, J = 12.9, 5.4 Hz, 1H), 4.44–4.38 (m, 1H), 4.11 (d, J = 10.0 Hz, 1H), 4.02 (d, J = 13.1 Hz, 2H), 3.79–3.76 (m, 1H), 3.64–3.61 (m, 1H), 3.55–3.48 (m, 1H), 3.15–3.13 (m, 6H), 3.00–2.87 (m, 3H), 2.86–2.72 (m, 4H), 2.71–2.64 (m, 2H), 2.35–2.21 (m, 3H), 2.01 (dd, J = 9.4, 4.2 Hz, 1H), 1.77 (dd, J = 31.9, 12.1 Hz, 4H), 1.54 (t, J = 15.3 Hz, 3H), 1.43–1.08 (m, 9H), 1.12–1.03 (m, 2H), 0.96 (dd, J = 14.7, 6.7 Hz, 3H).

[0631] LCMS m / z = 919 [M+1] +

[0632] Preparation of Example 38: 5-(4-(2-(4-(2-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)ethyl)piperidin-1-yl)ethyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0633]

[0634] a) Preparation of tert-butyl 4-((1,3-dioxolan-2-yl)methyl)piperidine-1-carboxylate

[0635] tert-Butyl 4-(2-oxoethyl)piperidine-1-carboxylate (1 g), ethylene glycol (0.27 g), p-toluenesulfonic acid (0.04 g), and toluene (10 mL) were added to a reaction flask and stirred at 100 °C overnight under an air atmosphere. After completion of the reaction, the resulting mixture was concentrated under reduced pressure, ethyl acetate (60 mL) was added, the organic layer was washed with water (20 mL × 3), and dried over anhydrous sodium sulfate. The solid was filtered off, and the filtrate was concentrated to obtain 980 mg of the title compound.

[0636] b) Preparation of 4-((1,3-dioxolan-2-yl)methyl)piperidine

[0637] tert-Butyl 4-((1,3-dioxolan-2-yl)methyl)piperidine-1-carboxylate (900 mg), zinc bromide (747 mg), and hexafluoroisopropanol (10 mL) were added to a reaction flask and stirred at room temperature overnight. After completion of the reaction, the solid was filtered off, and the reaction solution was concentrated to obtain 500 mg of the crude product, which was directly used in the next step.

[0638] c) Preparation of 5-(4-((4-((1,3-dioxolan-2-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0639] A solution of 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carbaldehyde (431 mg), 4-((1,3-dioxolan-2-yl)methyl)piperidine (200 mg) and titanium(IV) isopropoxide (664 mg) in 1,2-dichloroethane (5 mL) was stirred at room temperature for half an hour. Then sodium triacetoxyborohydride (495 mg) was added to the above mixture and stirring was continued at room temperature for 1.5 hours. After completion of the reaction, water (2 mL) was added to the resulting mixture to quench it, and the crude product was separated by reverse-phase column (column: C18, mobile phase: acetonitrile / water (0.1% NH3.H2O)) to give 150 mg of the title compound.

[0640] d) Preparation of 2-(1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidin-4-yl)acetaldehyde 5-(4-((4-((1,3-dioxolan-2-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (50 mg) was added to trifluoroacetic acid (1 mL) and stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was concentrated to obtain the crude product, which was directly used for the next step.

[0641] e) Preparation of 5-(4-(2-(4-(2-(((S)-2-(4-chlorophenyl))-3-((1R,6S)-5-((R)-5-methyl-7-oxo-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-4-yl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-3-oxopropyl)amino)ethyl)piperidin-1-yl)ethyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0642] (R)-4-((1S,6R)-5-((S)-3-amino-2-(4-chlorophenyl)propanoyl)-2,5-diazabicyclo[4.1.0]hept-2-yl)-5-methyl-5,8-dihydropyrido[2,3-d]pyrimidin-7(6H)-one (46 mg), 2-(1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidin-4-yl)acetaldehyde (40 mg), titanium(IV) isopropoxide (47 mg) and 1,2-dichloroethane (2 mL) were added to a reaction flask, and the mixture was stirred at room temperature under a nitrogen atmosphere for half an hour. Then sodium triacetoxyborohydride (35 mg) was added to the mixture, and stirring was carried out at room temperature under a nitrogen atmosphere for 2 hours. After completion of the reaction, the resulting mixture was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to give 5 mg of the title compound.

[0643] 1H NMR (400 MHz, DMSO-d6) δ 11.06 (s, 1H), 10.48 (d, J = 10.2 Hz, 1H), 8.20 (d, J = 7.8 Hz, 1H), 7.64 (d, J = 8.5 Hz, 1H), 7.39 (s, 2H), 7.35 (d, J = 4.0 Hz, 1H), 7.29 (d, J = 2.4 Hz, 2H), 7.25 - 7.18 (m, 1H), 5.06 (dd, J = 12.8, 5.4 Hz, 1H), 4.40 (dd, J = 15.6, 7.8 Hz, 1H), 4.26 - 4.06 (m, 1H), 4.05 - 3.94 (m, 3H), 3.90 - 3.82 (m, 1H), 3.79 - 3.54 (m, 2H), 3.22 - 3.18 (m, 1H), 3.18 - 3.11 (m, 2H), 2.93 (q, J = 12.4 Hz, 4H), 2.83 - 2.65 (m, 4H), 2.63 - 2.56 (m, 2H), 2.55 - 2.53 (m, 1H), 2.26 (t, J = 15.7 Hz, 2H), 2.14 - 1.92 (m, 4H), 1.88 - 1.65 (m, 6H), 1.62 - 1.44 (m, 4H), 1.43 - 1.35 (m, 1H), 1.24 (s, 3H), 1.11 (d, J = 4.2 Hz, 2H), 1.09 - 0.98 (m, 2H).

[0644] LCMS m / z = 905 [M+1]+

[0645] II. Active moiety

[0646] The specific structure of the positive reference compound used in the active experiment part is as follows:

[0647] (Positive reference MS170, J. Med. Chem. 2021, 64, 24, 18054 - 18081)

[0648] Experiment 1 In vitro enzyme activity test

[0649] 1. Materials and reagents

[0650] Envision model microplate reader (Molecular Devices)

[0651] White 384-well microplate (product number #264706, Thermo)

[0652] Main reagents included in the HTRK kinEASE STK kit (product number #62TKOPEC, Cisbio)

[0653] STK - Biotin Substrate

[0654] Streptavidin - XL665

[0655] Eu - labeled tyrosine kinase substrate antibody

[0656] 5× Enzyme Reaction Buffer

[0657] HTRF Detection Buffer

[0658] AKT1 (Catalog No. #01 - 101, Carna)

[0659] AKT2 (Catalog No. #01 - 102, Carna)

[0660] AKT3 (Catalog No. #PV3185, Invitrogen)

[0661] ATP 10 mM (Catalog No. #PV3227, Invitrogen)

[0662] DTT 1 M (Catalog No. #D5545, Sigma)

[0663] MgCl2 1 M (Catalog No. #M8266, Sigma)

[0664] 2. Experimental Procedures

[0665] 2.1 Reagent Preparation

[0666] Table 1 Components and Concentrations of the Reaction System for Kinases

[0667]

[0668] (1) 1× Kinase Reaction Buffer

[0669] 1 mL of 1× kinase reaction buffer for kinase AKT1 / 2 / 3 contains 200 μL of 5× kinase reaction buffer, 5 μL of 1 M MgCl2, 1 μL of 1 M DTT, and 794 μL of ultrapure water.

[0670] (2) 5× STK - Biotin Substrate and ATP Working Solution

[0671] The specific concentrations of STK - biotin substrate and ATP are shown in Table 1.

[0672] Dilute the substrate and ATP to 5 times the reaction concentration with 1× kinase reaction buffer.

[0673] (3) 5× Kinase Working Solution

[0674] The concentrations used for enzyme screening are shown in Table 1. Prepare the 5× enzyme working solution with 1× kinase reaction buffer.

[0675] (4) 4×Streptavidin-XL665 working solution

[0676] The concentration of streptavidin-XL665 in the reaction is shown in Table 1. Prepare the 4×streptavidin-XL665 working solution with the detection buffer.

[0677] (5) 4× Europium-labeled tyrosine kinase substrate antibody working solution

[0678] Dilute the europium-labeled tyrosine kinase substrate antibody 100-fold with the detection reaction buffer as the working solution.

[0679] 2.2 Experimental procedure

[0680] After all the reagents are prepared according to the above methods, except for the enzyme, equilibrate them to room temperature and then start adding samples.

[0681] a) First, dilute the compound stock solution (10 mM DMSO solution) with DMSO to a 100 μM compound solution, and then dilute it to a 2.5 μM compound working solution (containing 2.5% DMSO) with 1× kinase reaction buffer. Prepare a 2.5% DMSO solution with 1× kinase reaction buffer, and then dilute the 2.5 μM compound working solution with the 2.5% DMSO solution, and perform 7-fold serial dilutions by a factor of 4 for a total of 8 concentrations (2500 nM, 625 nM, 156 nM, 39 nM, 9.8 nM, 2.4 nM, 0.6 nM, 0.15 nM) of the compound working solution. Except for the control wells, add 4 μL of the diluted compound working solution to all reaction wells, and add 4 μL of the previously prepared 2.5% DMSO / kinase buffer solution to the control wells.

[0682] b) Add 2 μL of the previously prepared STK-biotin substrate solution (the amount of substrate used in enzyme screening is shown in Table 1) to all reaction wells.

[0683] c) Add 2 μL of the previously prepared enzyme solution (the amount of enzyme used is shown in Table 1) to all reaction wells except the negative wells, and fill the negative wells with 2 μL of the enzyme corresponding to 1× kinase reaction buffer to make up the volume. Seal the plate with a sealing film, mix well, and incubate at room temperature for 10 min to allow the compound and the enzyme to fully react and bind.

[0684] d) Add 2 μL of the ATP solution to all reaction wells to initiate the kinase reaction (the ATP concentration and reaction time in enzyme screening are shown in Table 1).

[0685] e) Start preparing the detection solution 5 min before the end of the kinase reaction. Prepare the streptavidin-XL665 and europium-labeled tyrosine kinase substrate antibody (1:100) detection solution (the concentration of the detection reagent in enzyme screening is shown in Table 1) with the detection buffer in the kit.

[0686] f) After the kinase reaction is completed, add 5 μL of diluted streptavidin-XL665 to all reaction wells. After mixing, immediately add the diluted europium-labeled tyrosine kinase substrate antibody detection solution.

[0687] g) Seal the plate and mix well. After reacting at room temperature for 1 h, detect the fluorescence signal using an ENVISION (Perkinelmer) instrument (stimulated at 320 nm, emitted at 665 nm and 615 nm). Calculate the inhibition rate of each well by using the fully active wells and background signal wells. Take the average value of duplicate wells. At the same time, perform the fitting of the half-inhibitory activity (IC 50 ) for each test compound using the professional graph analysis software PRISM 5.0.

[0688] Table 2: Experimental sample addition process table

[0689]

[0690]

[0691] 2.3 Data analysis

[0692] ER = fluorescence value at 665 nm / fluorescence value at 615 nm

[0693] Inhibition rate = (ER 阳性对照 - ER 样品 ) / (ER 阳性对照 - ER 阴性对照 ) * 100%

[0694] 3. Experimental results

[0695] The experimental results are shown in Table 3:

[0696] Table 3: AKT inhibitory activity

[0697]

[0698] Experiment 2 Detection of the degradation activity of compounds on target proteins in cells by NanoLuc method 1. Materials and reagents

[0699] HEK293 cells

[0700] DMEM medium (product number #SH30243.01, Hyclone)

[0701] Opti-MEM I medium (product number #31985-070, Gibco)

[0702] Fetal bovine serum (product number #10099-141C, Gibco)

[0703] FuGENE HD Transfection Reagent (Cat. No. E2312, Promega)

[0704] DMSO (catalog number #D2650-100 ml, Sigma)

[0705] pNLF1-N[CMV / Hygro] Vector (Cat. No. N1351, Promega)

[0706] Nano-Glo Luciferase Assay System (Cat. #N1110, Promega)

[0707] Trans5α Chemically Competent Cell (Cat.#CD201-01, TransGen)

[0708] LB sterile liquid medium (catalog number #B540111, Biotech)

[0709] UNIQ-500 Endotoxin-free Plasmid Mass Extraction Kit (Cat. No. B11249, Bio-Tech)

[0710] Ultra-micro digital pipette (Model #D300e, Tecan)

[0711] Cell counter (model #RigelS2, Countstar)

[0712] Multifunctional microplate reader (Model #SPARK, Tecan)

[0713] Nucleic acid quantitative analyzer (model #NanoDrop One, Thermo)

[0714] 2. Experimental Procedure

[0715] 2.1 Construction and amplification of target plasmid

[0716] Using molecular biology techniques, the complete nucleic acid sequences encoding the proteins of AKT1 (NM_005163.2), AKT2 (NM_001626.6), and AKT3 (NM_005465.7) were cloned into the pNLF1-N[CMV / Hygro] plasmid. The restriction enzyme sites selected were: EcoRI at the 5' end and XbalI at the 3' end. After sequencing confirmed that the target sequence was correctly inserted into the pNLF1-N[CMV / Hygro] plasmid (the target plasmid), the target plasmid was transformed into Trans5α Chemically Competent Cell. Then, the target plasmid was amplified using LB sterile liquid medium (220 rpm, 16 - 18 h). After amplification, the target plasmid was purified using the UNIQ-500 endotoxin-free plasmid large extraction kit, and the concentration and purity of the target plasmid were detected using NanoDrop One.

[0717] 2.2 Transfection of the target plasmid

[0718] a) Dilute the target plasmids (AKT1, AKT2, AKT3) to 10 ng / μL using TE Buffer.

[0719] b) Adjust the density of HEK293 cells to 2E5 / mL using DMEM medium.

[0720] c) Dilute 2 μg of plasmid complex (10 ng target plasmid and 1990 ng pcDNA3.1(+) empty plasmid) in 200 μL of Opti-

[0721] MEM I medium and mix well.

[0722] d) Add 6 μL of FuGENE HD Transfection Reagent to the Opti-MEM I medium containing the plasmid, invert and mix 5 - 10 times, and incubate at room temperature for 20 min.

[0723] e) Add the incubated plasmid / transfection reagent mixture to 5 mL of HEK-293 cells, invert and mix 5 times, then transfer to a culture flask and culture in a 37°C, 5% CO2 incubator for 12 hours.

[0724] 2.3 Compound treatment

[0725] The cells in the culture flask were digested, centrifuged, and counted routinely. The HEK-293 cells transfected with the target plasmid were seeded into a white-bottom opaque 96-well plate at a density of 10,000 cells per well (100 μL of medium per well). After 24 h, different concentrations of the compound were added to the cells using an ultra-micro digital pipettor for the corresponding time (4 h to 24 h).

[0726] 2.4 Signal Detection and Data Analysis

[0727] a) Thaw the Substrate and Buffer in the Nano-Glo Luciferase Assay System kit overnight in a 4°C refrigerator. According to the required reagent volume for the experiment, mix 25 μL of Substrate with 1.25 mL of Buffer in proportion (the mixture is the detection working solution. After thawing, first pipette and mix the Substrate evenly, then add it to the Buffer and mix well). The prepared reagent is stored in a 4°C refrigerator for later use.

[0728] b) Equilibrate the detection working solution to room temperature before detection, and place the cells at room temperature for 10 min.

[0729] c) Add the detection working solution with the same volume as the cell culture medium to the 96-well plate and mix on a microplate oscillator for 3 min.

[0730] d) Use a multi-functional microplate reader to detect the chemiluminescence signal of each well.

[0731] e) Use Graphpad Prism software to calculate DC 50 (50% degradation concentration of the compound) and D MAX (maximum degradation degree of the target protein).

[0732] 3. Experimental Results

[0733] The experimental results are shown in Table 4

[0734] Table 4 Degradation Activity of Compounds on Target Proteins in Cells Detected by NanoLuc Method

[0735]

[0736]

[0737]

[0738]

[0739] Experiment 3: AKT1 / 2 / 3 Protein Degradation ICW (In-Cell Western) Experiment on PC-3 Cells

[0740] On the first day, inoculate PC-3 cells (human prostate cancer cells, culture medium: F12K + 10% FBS + 1% P / S), purchased from ATCC (product number: CRL-1435). Seed the above cells in the logarithmic growth phase at a cell density of 3000 cells / well in a 384-well plate and culture overnight in an incubator at 37°C and 5% CO2.

[0741] On the second day, incubate the compounds. Use an Echo 665 Liquid Handler to add the compounds, adding gradient-diluted compounds at 40 μl / well (starting concentration 10 μM, 3-fold serial dilution, a total of 10 doses, 0.1% DMSO), and culture in an incubator at 37°C and 5% CO2 for 24 h.

[0742] On the third day, centrifuge the plates for 1 minute. Use an Apricot fully automatic pipetting device, the Personal Pipettor 384-channel pipetting workstation, to add 40 μl / well of 8% PFA paraformaldehyde fixative and incubate at room temperature for 20 min. Remove the supernatant, wash twice with 40 μl / well of DPBS, then permeabilize the cells with 100% cold methanol and incubate at room temperature for 10 min. Remove the supernatant, wash once with 40 μl / well of DPBS, add 40 μl / well of Li-Cor blocking buffer, and incubate at room temperature for 1 h. Remove the supernatant, add 20 μl / well of primary antibody solution (dilute the antibody with the blocking buffer, AKT1 Rabbit mAb (CST, catalog number: 75692S) diluted 1:500; AKT2 Rabbit mAb (CST, catalog number: 3063S) diluted 1:1000, AKT3 Rabbit mAb (CST, catalog number: 14982S) diluted 1:200), and incubate overnight at 4°C.

[0743] On the fourth day, discard the supernatant, soak with 40 μl / well of TBST (0.05% Tween-20 in TBS) for 5 min each time, and wash a total of 3 times. Then, add 20 μl / well of secondary antibody solution (Alexa Fluor 488 goat anti-rabbit 1:1000 and Hoechst 1:5000 diluted with the blocking buffer), and incubate at room temperature for 1 h. Discard the supernatant, soak with 40 μl / well of PBST (0.05% Tween-20 in PBS) for 5 min each time, and wash a total of 3 times. Centrifuge the plates at 1000 rpm for 1 min, then use an Operetta high-content cell analyzer for detection. Use GraphPad Prism 5 software to fit the dose-response curve with log(inhibitor) vs. response - Variable slope, and respectively obtain the DC 50 value.

[0744] The compounds of the relevant examples have an activity of <100 μM.

[0745] Experiment 4: MDA-MB-468 cell proliferation inhibition experiment

[0746] The MDA-MB-468 human breast cancer cell line (purchased from KBP Biosciences, Cat: CBP60387) was cultured in DMEM medium containing 10% FBS (Fetal bovine serum). Cells in the logarithmic growth phase were seeded in 96-well plates (purchased from PerkinElmer, Cat: 6005181) at a density of 3000 cells / well. Additionally, a G0 control plate for detection on the second day was set up. The seeded cell plates were placed in an incubator at 37°C and 5% CO2 for 24 h.

[0747] On the first day of the experiment, the cell plates were taken out, and compounds were added using an automatic liquid dispenser D300e (Tecan). The initial concentration of the compounds was 30 mM, and they were serially diluted 3-fold, with a total of 9 gradients. The final concentration of DMSO in each well was 0.3%. After adding the drugs, the cell plates were placed in an incubator at 37°C and 5% CO2 for 120 h. Meanwhile, the G0 control plate was taken out of the incubator, and 50 μl of Cell Counting-Lite TM 2.0 solution (purchased from Vazyme) was added to each well, and the fluorescence signal was measured on an Envision (PerkinElmer) according to the operating procedure in the instruction manual.

[0748] On the sixth day of the experiment, the cell plates were taken out, and 50 μl of Cell Counting-Lite TM 2.0 solution (purchased from Vazyme) was added to each well, and the fluorescence signal was measured on an Envision (PerkinElmer) according to the operating procedure in the instruction manual. The dose-response curve was fitted using GraphPad Prism 5 software: log(inhibitor) vs. response - Variable slope, and the absolute GI 50 value of the compound on cell proliferation inhibitory activity was obtained. The formula for viability is:

[0749]

[0750] Where:

[0751] Signal value of the test substance: mean fluorescence signal of the cell + medium + compound group;

[0752] Signal value of the G0 group: mean fluorescence signal of the cell + medium group detected on the first day;

[0753] Signal value of the negative control group: mean fluorescence signal of the cell + medium group detected on the sixth day.

[0754] For the specific results, see Table 5

[0755] Table 5 Results of the inhibitory experiment on the proliferation of MDA-MB-468 cells

[0756] Example Title Compound <![CDATA[GI 50 (nM) <!-- 84 -->]]> Example 3 101.08 Yangshen Compound 322.6

[0757] Experiment 5: Pharmacokinetics experiment in mice

[0758] (1) Prescription preparation of the compound:

[0759] Mixed solvent: DMSO:Solutol HS: physiological saline = 5:10:85 (V / V / V)

[0760] Process of drug preparation:

[0761] Weigh the required weight of the test substance and place it in a brown glass bottle. Add DMSO accounting for 5% of the total volume of the solvent, mix well, then add Solutol HS accounting for 10% of the total volume of the solvent, mix well, and finally add physiological saline accounting for 85% of the total volume of the solvent. Ultrasonic until the solution is clear to obtain a clear and transparent solution with the required concentration.

[0762] (2) Experimental animals: Mice, strain ICR, source: Hunan Slack Jingda Experimental Animal Co., Ltd., age: 6 - 8 weeks, male.

[0763] (3) Experimental protocol:

[0764] The experimental animals are housed in the animal house of the Experimental Animal Center of Hunan University of Chinese Medicine. The animals are kept in rat cages. The animal house has good ventilation and is equipped with air conditioning. The temperature is maintained at 20 - 26 °C and the humidity is maintained at 40% - 70%. A time-controlled lighting system is used to provide a 12-hour light / 12-hour dark day-night cycle. During the study, the animal cages and water bottles will be replaced regularly. After normal feeding for at least 3 days, mice with good physical signs can be selected for this experiment after veterinary inspection. Each mouse is marked with a tail number. The animal experiment protocol is shown in Table 1 for details.

[0765] Table 1 Animal experiment protocol table

[0766]

[0767] After weighing the body weight, calculate the theoretical dosing volume for each mouse according to the following formula.

[0768]

[0769] One day before the experiment, the mice are fasted overnight (>12 hours), can drink water freely, and are fed 4 hours after dosing.

[0770] On the day of the experiment, the mice in the dosing group were respectively gavaged with 10 mg / kg of the dosing solution. After dosing, about 0.1 mL of blood was collected from the orbital cavity of the mice at each time point and placed in an EDTA-K2 anticoagulant tube. After gently inverting up and down to fully mix the anticoagulant (EDTA-2K) with the blood, it was immediately placed on wet ice and centrifuged rapidly within 1 h after blood collection to separate plasma, and the centrifugation conditions were set at 4 °C, 6800 g, and 6 minutes. After centrifugation, the plasma was loaded into a labeled EP tube and stored in a -20 °C refrigerator within 1 hour for biological sample analysis. An LC-MS / MS analytical method for determining the compound concentration in mouse plasma was established and used to determine the compound concentration in the biological samples obtained in this experiment. The non-compartmental model in Phoenix WinNonlin 8.3 was used to calculate the pharmacokinetic parameters.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, with the specific structure as follows: B—L—D(I) Wherein, L is selected from —Ak1—C1—Ak2—C2—Ak3— or —Ak4—Cy1—Ak5—Cy2—Ak6—; wherein Ak1, Ak2, and Ak3 are independently selected from or a bond; C1 and C2 are independently selected from C 1-12 alkyl, C 2-12 alkenyl or C 2-12 alkynyl or a bond, provided that C1 and C2 are not simultaneously a bond. The C 1-12 alkyl, C 2-12 alkenyl or C 2-12 alkynyl is optionally substituted by one or more C 1-4 alkyl or halogen; wherein Ak4, Ak5, and Ak6 are independently selected from optionally C substituted with one or more oxygen, C 1-4 alkyl or halogen substituted C 1-12 alkyl or bond; Cy1 and Cy2 are independently selected from a 3- to 12-membered heterocycle, a 3- to 12-membered cycloalkyl, a 6- to 10-membered aryl or a bond, and Cy1 and Cy2 are not simultaneously a bond. The 3- to 12-membered heterocycle, 3- to 12-membered cycloalkyl, 6- to 10-membered aryl are optionally substituted by one or more halogens or C 1-4 alkyl groups of; B is Wherein X is selected from C or N; R 1 is halogen or hydrogen; R 2 selected from H, CN, C 1-6 alkyl or C 1-6 alkoxy, wherein the C 1-6 alkyl or C 1-6 alkoxy is optionally substituted by halogen, OH, CN; G is a 5- to 12-membered saturated heterocycle containing 1 to 2 nitrogen atoms optionally substituted by C 1-4 alkyl; R 3 is NH, or does not exist; D is selected from the structures shown below: When B is , L is not —Ak1—C1—Ak2—C2—Ak3—.

2. The compound of formula (I) according to claim 1, wherein, L is selected from C 1-12 alkyl, C 2-12 alkenyl or C 2-12 alkynyl, wherein the C 1-12 alkyl, C 2-12 alkenyl or C 2-12 alkynyl is optionally substituted by one or more C 1-4 alkyl or halogen, or —Ak4—Cy1—Ak5—Cy2—Ak6, wherein Ak4, Ak5, and Ak6 are independently selected from optionally C-alkyl or a bond substituted by one or more oxygens 1-12 alkyl or bond, Cy1 and Cy2 are independently selected from optionally 3- to 12-membered heterocycles or bonds optionally substituted by one or more halogens, C 1-4 alkyl, and Cy1 and Cy2 are not simultaneously bonds; Preferably, L is selected from optionally C substituted by one or more C 1-4 alkyl or halogen-substituted C 1-12 alkyl or C 2-12 alkynyl, or —Ak4—Cy1—Ak5—Cy2—Ak6; wherein Ak4, Ak5, and Ak6 are independently selected from C alkyl or a bond substituted by one or more oxygens 1-10 ; further, Ak4, Ak5, and Ak6 are independently selected from C 1-10 alkyl or a bond Cy1 and Cy2 are independently selected from 4- to 9-membered heterocycles or bonds, and Cy1 and Cy2 are not both bonds at the same time; Preferably, L is selected from optionally C substituted by one or more methyl groups 1-12 alkyl or C 2-12 alkynyl, more preferably optionally C substituted by one or more methyl groups 8-12 alkyl or C 8-12 alkynyl, further preferably C 8-12 alkyl or C 8-12 alkynyl.

3. The compound of formula (I) according to claim 2, wherein, Cy1 and Cy2 are independently selected from 4- to 9-membered heterocycles or bonds, and Cy1 and Cy2 are not both bonds at the same time; Preferably, Cy1 and Cy2 are independently selected from 4- to 9-membered heterocycles containing 1 to 2 N atoms or bonds, and Cy1 and Cy2 are not both bonds at the same time; Preferably, Cy1 and Cy2 are independently selected from azetidinyl, pyrrolidinyl, piperidine, piperazine, cyclopropylazetidinyl, cyclopropylpyrrolidinyl, cyclopropylpiperidinyl, cyclopropylpiperazinyl, cyclobutylazetidinyl, cyclobutylpyrrolidinyl, cyclobutylpiperidinyl, cyclobutylpiperazinyl, cyclopentylazetidinyl, cyclopentylpyrrolidinyl, cyclopentylpiperidinyl, cyclopentylpiperazinyl, cyclohexylazetidinyl, cyclohexylpyrrolidinyl, azetidinylazetidinyl, azetidinylpyrrolidinyl, azetidinylpiperidinyl, azetidinylpiperazinyl, pyrrolidinylazetidinyl, pyrrolidinylpyrrolidinyl, pyrrolidinylpiperidinyl, pyrrolidinylpiperazinyl, cyclobutylspiroazetidinyl, cyclobutylspiropyrrolidinyl, cyclobutylspiropiperidinyl, cyclopentylspiroazetidinyl, cyclopentylspiropyrrolidinyl, cyclohexylspiroazetidinyl, azetidinylspiroazetidinyl, azetidinylspiropyrrolidinyl, azetidinylspiropiperidinyl, pyrrolidinylspiroazetidinyl, pyrrolidinylspiropyrrolidinyl, piperidinylspiroazetidinyl, cyclobutylspiropiperazinyl, azetidinylspiropiperazinyl, pyrrolidinylspiropiperazinyl, piperidinylspiropiperazinyl or bonds, and Cy1 and Cy2 are not both bonds at the same time; Preferably, Cy1 and Cy2 are independently selected from piperidine, piperazine or cyclobutylspiropiperidinyl or bonds, and Cy1 and Cy2 are not both bonds at the same time.

4. The compound of formula (I) according to claim 1, wherein L is selected from the following structures: where the left side of L is connected to B and the right side is connected to D; Preferably, L is selected from the following structures: Preferably, L is 5. The compound of formula (I) according to claim 1, wherein B is Wherein X is selected from C or N; R 1 is chlorine or hydrogen; R 2 is a C1-C6 alkyl group, wherein the C1-C6 alkyl group is optionally substituted by OH; G is an optionally methyl-substituted 5- to 12-membered saturated heterocycle containing 1 to 2 nitrogen atoms; R 3 is NH, or does not exist.

6. The compound of formula (I) according to claim 5, wherein R 2 is methyl, ethyl or —(CH2)2OH, preferably, R 2 is methyl or ethyl; R 1 wherein Cl is chlorine and G is any of the following groups optionally substituted with methyl: Preferably, G is selected from the following groups: Preferably, G is selected from the following groups:

7. The compound as shown in claim 1, wherein B is selected from the following structures: Preferably, B is selected from the following structures: Preferably, B is selected from the following structures: Preferably, B is selected from the following structures:

8. The compound of formula (I) as shown in claim 1, wherein D is selected from the following structures: Preferably, D is selected from the following structures: Preferably, D has the following structure 9. A compound of formula (II), formula (III), formula (IV), formula (V) or formula (VI) or a pharmaceutically acceptable salt thereof, the specific structures are as follows: L1 is selected from —Ak1—C1—Ak2—C2—Ak3— or —Ak4—Cy1—Ak5—Cy2—Ak6—; Wherein Ak1, Ak2, Ak3 are independently selected from Or a bond; C1 and C2 are independently selected from C 1-12 alkyl groups of 2-12 alkenyl groups of 2-12 alkynyl groups of 1-12 or a bond, provided that C1 and C2 are not simultaneously a bond. The C 2-12 alkyl groups of 2-12 alkenyl groups of 1-4 alkynyl groups of are optionally substituted by one or more C 1-4 alkyl groups of 1-12 or a halogen; Cy1 and Cy2 are independently selected from 3- to 12-membered heterocycles, 3- to 12-membered cycloalkyl groups, 6- to 10-membered aryl groups or a bond, provided that Cy1 and Cy2 are not simultaneously a bond. The 3- to 12-membered heterocycles, 3- to 12-membered cycloalkyl groups, 6- to 10-membered aryl groups are optionally substituted by one or more halogens or C 1-4 alkyl groups of D1 is selected from the structures shown below: L2 is selected from —Ak1—C1—Ak2—C2—Ak3— or —Ak4—Cy1—Ak5—Cy2—Ak6—; wherein Ak1, Ak2, Ak3 are independently selected from or a bond; C1 and C2 are independently selected from C 1-12 alkyl groups of 2-12 alkenyl groups of 2-12 alkynyl groups of 1-12 or a bond, provided that C1 and C2 are not simultaneously a bond. The C 2-12 alkyl groups of 2-12 alkenyl groups of 1-4 alkynyl groups of are optionally substituted by one or more C 1-4 alkyl groups of 1-12 or a halogen; Cy1 and Cy2 are independently selected from 3- to 12-membered heterocycles, 3- to 12-membered cycloalkyl groups, 6- to 10-membered aryl groups or a bond, provided that Cy1 and Cy2 are not simultaneously a bond. The 3- to 12-membered heterocycles, 3- to 12-membered cycloalkyl groups, 6- to 10-membered aryl groups are optionally substituted by one or more halogens or C 1-4 alkyl groups of D2 is selected from the structures shown below: wherein, L3 is —Ak4—Cy1—Ak5—Cy2—Ak6—; Wherein Ak4, Ak5, and Ak6 are independently selected from optionally C substituted by one or more oxygen, C 1-4 alkyl or halogen substituted C 1-12 alkyl or bond; Cy1 and Cy2 are independently selected from a 3- to 12-membered heterocyclic ring, a 3- to 12-membered cycloalkyl group, a 6- to 10-membered aryl group or a bond, provided that Cy1 and Cy2 are not simultaneously a bond, wherein the 3- to 12-membered heterocyclic ring, the 3- to 12-membered cycloalkyl group, the 6- to 10-membered aryl group are optionally substituted by one or more halogen atoms or C 1-4 alkyl groups; D3 is selected from the structures shown below: wherein, L4 is —Ak4—Cy1—Ak5—Cy2—Ak6—; Wherein Ak4, Ak5, and Ak6 are independently selected from optionally C substituted by one or more oxygen, C 1-4 alkyl or halogen substituted C 1-12 alkyl or bond; Cy1 and Cy2 are independently selected from 3- to 12-membered heterocycles, 3- to 12-membered cycloalkyls, 6- to 10-membered aryls or a bond, provided that Cy1 and Cy2 are not simultaneously a bond, wherein the 3- to 12-membered heterocycles, 3- to 12-membered cycloalkyls, 6- to 10-membered aryls are optionally substituted by one or more halogens or C 1-4 alkyl groups; D4 is selected from the structures shown below: wherein, L5 is —Ak4—Cy1—Ak5—Cy2—Ak6—; wherein Ak4, Ak5, and Ak6 are independently selected from optionally C substituted with one or more oxygen, C 1-4 alkyl or halogen substituted C 1-12 alkyl or bond; Cy1 and Cy2 are independently selected from a 3- to 12-membered heterocyclic ring, a 3- to 12-membered cycloalkyl group, a 6- to 10-membered aryl group or a bond, and Cy1 and Cy2 are not simultaneously a bond, wherein the 3- to 12-membered heterocyclic ring, the 3- to 12-membered cycloalkyl group, and the 6- to 10-membered aryl group are optionally substituted with one or more halogen atoms or C 1-4 alkyl groups of; D5 is selected from the structures shown below:

10. A compound having the following structure or a pharmaceutically acceptable salt thereof,

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