Compounds and methods for degrading casein kinase 1 alpha

Abstract

Provided are a novel compound represented by formula (I), which degrades CK1 [alpha] (casein kinase 1 [alpha]), and a method for degrading CK1 [alpha] or preventing or treating leukemia by administering the compound to a subject in need thereof.

Description

Background Art

[0001] Casein kinase 1α (CK1α) is a serine / threonine protein kinase that is involved in various cellular pathways and functions, including Wnt signaling, NF-κB signaling, the p53 pathway, autophagy, and the cell cycle. Inhibition of CK1α or reduction of its expression levels has been shown to induce cell death in several types of cancer, such as acute myeloid leukemia (AML) and diffuse large B-cell lymphoma (DLBCL).

[0002] For example, it has been reported that CK1α inhibition causes reduced Rps6 phosphorylation and activation of p53, leading to the selective elimination of leukemic cells, revealing CK1α as a therapeutic target for the treatment of AML (Jaras et al., J. Exp. Med. 2014 Vol. 211 No. 4 605-612). In addition, it has been reported that in AML and multiple myeloma (MM), inhibition of CK1α, which acts through activation of the p53 pathway, shows promising preclinical activity (Janovska et al., Int. J. Mol. Sci. 2020, 21, 9026). Specifically, it has been previously discovered that CK1α controls signaling pathways involved in proliferation, survival, and stress in MM (Manni et al., Journal of Hematology and Oncology (2017) 10: 157). In addition, it has been reported that CK1α is overexpressed in AML patients and acts as a negative element in the prognosis of AML patients, so CK1α inhibits p53 downstream of MDM2-mediated autophagy and apoptosis, and the targeting of CK1α and autophagy can provide a therapeutic opportunity for treating AML (Xu et al., ONCOLOGY REPORTS 44:1895-1904, 2020). CK1α has also been found to dominate antigen receptor-induced NF-κB activation and human lymphoma cell survival.

[0003] Therefore, CK1α is known in the art as a target for preventing or treating blood cancers (such as leukemia, lymphoma and myeloma). Therefore, there is a need to develop CK1α degraders or inhibitors that can be used to prevent or treat blood cancers (such as leukemia, lymphoma and myeloma). Summary of the Invention

[0004] The present disclosure provides novel compounds or pharmaceutically acceptable salts, enantiomers, stereoisomers or prodrugs thereof, wherein the compounds are represented by the following formula (I).

[0005]

[0006] The compound of formula (I) or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof is used for reducing CK1α protein levels and for treating or preventing blood cancer.

[0007] In formula (I), in some embodiments, R 1 is: C1-C8 alkyl, C3-C8 alkenyl, C3-C8 alkynyl, C3-C8 cycloalkyl or C5-C 10 Bicycloalkyl, each optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, NH2, C1-C3 haloalkyl, optionally substituted C6-C 10 Aryl, C1-C3 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C5-C 10 bicycloalkyl or optionally substituted C3-C6 heterocycloalkyl.

[0008] In some embodiments, X is: a bond; or -(CH2) n –, optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, C1-C3 haloalkyl, C3-C6 cycloalkyl or C1-C6 alkyl, wherein the C1-C6 alkyl group may form, together with the atoms to which it is attached, a C3-C6 spiroalkyl ring, and wherein –(CH2) n – The group may contain 0-1 double or triple bonds.

[0009] In some embodiments, n is an integer from 1 to 6.

[0010] In some embodiments, A is: a bond; C4-C8 cycloalkyl, C5-C 10 Bicycloalkyl, C5-C 10 Heterobicycloalkyl or C3-C8heterocycloalkyl, each optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, CN, C1-C3 haloalkyl, C1-C3 alkyl or C3-C5 cycloalkyl.

[0011] In some embodiments, Y is a bond or -(CH2) m –.

[0012] In some embodiments, m is an integer of 1 or 2.

[0013] In some embodiments, Z is H or halogen.

[0014] In some embodiments, R 2 Yes:-NR 12 R 5 、-NHC(O)R 6 、-NHC(O)NR 8 (R 9 )、-C(O)NHR5 , five-membered or six-membered heteroaryl or C3-C8 heterocycloalkyl ring or C5-C 10 Heterobicycloalkyl ring, optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, CN, C1-C3 haloalkyl, C1-C3 alkyl, or C3-C5 cycloalkyl.

[0015] In some embodiments, R 5 is: H, C1-C5 alkyl, C3-C7 cycloalkyl, five-membered or six-membered heteroaryl, C5-C 10 Bicycloalkyl, (C3-C6 cycloalkyl)-C1-C3 alkyl, (C3-C6 heterocycloalkyl)-C1-C3 alkyl, (C6-C 10 (C1-C5-heteroaryl)-C1-C3 alkyl, or C3-C7 heterocycloalkyl, each optionally substituted with one or more halogen, CN, OH, O-(C1-C3 haloalkyl), O-(C1-C3 alkyl), C1-C3 haloalkyl, CH3SO2-, optionally substituted C3-C6 cycloalkyl, optionally substituted C3-C7 heterocycloalkyl, or C(O)NR 8 (R 9 ).

[0016] In some embodiments, R 6 is: C1-C6 alkyl, C3-C6 cycloalkyl or C3-C6 heterocycloalkyl, each optionally substituted with one or more halogen, C1-C3 haloalkyl, optionally substituted C3-C6 cycloalkyl or NR 8 (R 9 ).

[0017] In some embodiments, R 8 and R 9 are all independently H, substituted C1-C3 alkyl, or taken together with the nitrogen to which they are attached to form a 4-6 membered heterocyclic ring optionally substituted with one or more halogen or C1-C3 haloalkyl.

[0018] In some embodiments, R 12 is: H, C1-C5 alkyl, or the nitrogen to which it is attached and R 5 Together they form (i) a C3-C8 cycloalkyl ring or a heterocycloalkyl ring or (ii) a C5-C 10 A bicycloalkyl ring or a heterobicycloalkyl ring, each optionally substituted with one or more halogen, C1-C3 haloalkyl, OH, O-(C1-C3 alkyl), O-(C1-C3 haloalkyl) or CN.

[0019] In some embodiments, each stereocenter in the compound of Formula (I) is independently an R-enantiomer, an S-enantiomer, or a mixture of an R-enantiomer and an S-enantiomer.

[0020] In some embodiments, each double bond in the compound of Formula (I) is independently cis or trans.

[0021] In some embodiments, a compound or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof is provided, wherein the compound is represented by formula (Ia):

[0022]

[0023] In Formula (Ia), in some embodiments, X is a bond or -(CH2) n –.

[0024] In some embodiments, R 1 is: C1-C8 alkyl or C3-C8 cycloalkyl, each optionally substituted with one or more halogen, OH, NH2, C1-C3 alkyl, C1-C3 haloalkyl, O-(C1-C3 alkyl), optionally substituted C6 aryl, optionally substituted C3-C8 cycloalkyl, optionally substituted C3-C6 heterocycloalkyl or optionally substituted C6-C8 bicycloalkyl.

[0025] In some embodiments, A is: C4-C6 cycloalkyl, C4-C6 heterocycloalkyl, or C5-C7 bicycloalkyl, each optionally substituted with one or more halogen, OH, C1-C3 alkyl, or C1-C3 haloalkyl.

[0026] In some embodiments, Y is a bond or -CH2-.

[0027] In some embodiments, R 5 is: H, C1-C5 alkyl, C3-C6 cycloalkyl, 6-membered heteroaryl, C5 bicycloalkyl, (C3-C4 cycloalkyl)-C1-C2 alkyl, (C3-C4 heterocycloalkyl)-C1-C2 alkyl, (C6 aryl)-C1-C2 alkyl, (C5 heteroaryl)-C1-C2 alkyl or C3-C4 heterocycloalkyl, each optionally substituted with one or more halogen, CN, OH, O-(C1-C3 haloalkyl), O-(C1-C3 alkyl), C1-C3 haloalkyl, CH3SO2- or C(O)NR 8 (R 9 ).

[0028] In some embodiments, R 12 is H, C1-C5 alkyl, or the nitrogen to which it is attached and R 5Together they form a C3-C8 heterocycloalkyl ring optionally substituted with one or more halogen or C1-C3 haloalkyl.

[0029] In some embodiments, a compound or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof is provided, wherein the compound is represented by formula (Ib):

[0030]

[0031] In Formula (Ib), in some embodiments, X is a bond or -(CH2) n –.

[0032] In some embodiments, R 1 is: C3-C6 alkyl, optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, NH2, C1-C3 haloalkyl.

[0033] In some embodiments, A is: a bond; or C4-C6 cycloalkyl, optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, NH2, C1-C3 haloalkyl.

[0034] In some embodiments, Y is a bond.

[0035] In some embodiments, R 6 is optionally replaced with NR 8 (R 9 ) of a C1-C6 alkyl group.

[0036] In some embodiments, a compound or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof is provided, wherein the compound is represented by formula (Ic):

[0037]

[0038] In Formula (Ic), in some embodiments, X is a bond or -(CH2) n –.

[0039] In some embodiments, R 1 is: C1-C6 alkyl, optionally substituted with optionally substituted C3-C8 cycloalkyl or optionally substituted C3-C6 heterocycloalkyl.

[0040] In some embodiments, A is: a bond; C4-C6 cycloalkyl, C4-C6 heterocycloalkyl, or C5-C7 bicycloalkyl, each optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, NH2, or C1-C3 haloalkyl.

[0041] In some embodiments, Y is a bond.

[0042] In some embodiments, R 5 is H or C1-C3 alkyl.

[0043] In some embodiments, a compound or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof is provided, wherein the compound is represented by formula (Id):

[0044]

[0045] In Formula (Id), in some embodiments, R 1 is a C1-C3 alkyl group optionally substituted with an optionally substituted C3-C6 cycloalkyl group.

[0046] In some embodiments, A is optionally substituted C4-C6 cycloalkyl.

[0047] In some embodiments, R 2 It is: C1-C4 alkyl or (C3-C4 cycloalkyl)-C1-C2 alkyl, each optionally substituted with one or more halogen, CN, OH, O-(C1-C3 haloalkyl), O-(C1-C3 alkyl), C1-C3 haloalkyl or CH3SO2-.

[0048] In Formula (I), in some embodiments, each X and Y is a bond; A is a C4-C6 cycloalkyl group; R 1 is C1-C3 alkyl optionally substituted with optionally substituted C3-C6 cycloalkyl; and R 2 It is a 5-membered heteroaryl group.

[0049] In some embodiments, the compound is selected from Table A below.

[0050] [Table A]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056]

[0057]

[0058]

[0059]

[0060]

[0061]

[0062]

[0063]

[0064]

[0065]

[0066]

[0067]

[0068]

[0069]

[0070]

[0071]

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102]

[0103]

[0104]

[0105]

[0106]

[0107]

[0108]

[0109]

[0110]

[0111]

[0112]

[0113]

[0114]

[0115]

[0116]

[0117]

[0118]

[0119]

[0120]

[0121] In some embodiments, the compounds of the present disclosure are used to degrade and / or reduce casein kinase 1 alpha (CK1 alpha).

[0122] In some embodiments, the compounds of the present disclosure are used to inhibit casein kinase 1α (CK1α) activity.

[0123] In some embodiments, the compounds of the present disclosure are used to prevent or treat blood cancers.

[0124] In some embodiments, the compounds of the present disclosure are used in the preparation of a medicament for the treatment of blood cancer.

[0125] In some embodiments, the present disclosure provides a pharmaceutical composition comprising: a compound of the present disclosure (represented by Formula (I), Formula (Ia), Formula (Ib) or Formula (Ic)), or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof; and optionally a pharmaceutically acceptable excipient or carrier.

[0126] In some embodiments, the pharmaceutical composition is used to degrade and / or reduce casein kinase 1 alpha (CK1 alpha).

[0127] In some embodiments, the pharmaceutical composition is used to inhibit casein kinase 1α (CK1α) activity.

[0128] In some embodiments, the pharmaceutical composition is used to prevent or treat blood cancer.

[0129] In some embodiments, the blood cancer is leukemia, lymphoma, or myeloma.

[0130] The present disclosure also provides a method of degrading and / or reducing casein kinase 1α (CK1α) in a subject in need thereof, the method comprising administering to the subject an effective amount of the pharmaceutical composition of the present disclosure.

[0131] The present disclosure also provides a method of inhibiting casein kinase 1α (CK1α) activity in a subject in need thereof, the method comprising administering to the subject an effective amount of the pharmaceutical composition of the present disclosure.

[0132] The present disclosure also provides a method for preventing or treating blood cancer in a subject in need thereof, comprising administering to the subject an effective amount of the pharmaceutical composition of the present disclosure.

[0133] In some embodiments, the blood cancer is leukemia, lymphoma, or myeloma.

[0134] In some embodiments, the leukemia is selected from the group consisting of acute myeloid leukemia (AML), acute lymphoblastic leukemia or acute lymphoblastic leukemia (ALL), T-cell acute lymphoblastic leukemia (T-ALL), B-cell acute lymphoblastic leukemia (B-ALL), chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), T-cell prolymphocytic leukemia (T-PLL), large granular lymphocytic leukemia, adult T-cell leukemia, chronic eosinophilic leukemia (CEL) and myelodysplastic syndrome (MDS).

[0135] In some embodiments, the lymphoma is selected from the group consisting of Hodgkin lymphoma, diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, small lymphocytic lymphoma (SLL), mantle cell lymphoma (MCL), marginal zone lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, primary central nervous system (CNS) lymphoma, and peripheral T-cell lymphoma.

[0136] In some embodiments, the myeloma is selected from the group consisting of multiple myeloma, light chain myeloma, non-secretory myeloma, solitary plasmacytoma, extramedullary plasmacytoma, monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM), immunoglobulin D (IgD) myeloma, and immunoglobulin E (IgE) myeloma. DETAILED DESCRIPTION

[0137] Having benefited from the teachings provided in the foregoing description, those skilled in the art of the disclosed compositions and methods will recognize many modifications and other embodiments disclosed herein. Therefore, it will be understood that the disclosure is not limited to the specific embodiments disclosed, and that modifications and other embodiments are intended to be included within the scope of the appended claims. Those skilled in the art will recognize many variations and adaptations of the aspects described herein. Such variations and adaptations are intended to be included within the teachings of the present disclosure and are encompassed by the claims herein.

[0138] Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

[0139] Any enumerated method may be practiced in the order of events recited, or in any other order that is logically feasible. That is, unless expressly stated otherwise, it is not intended that any method or aspect described herein be construed as requiring that its steps be performed in a particular order. Therefore, in the absence of a specific limitation of steps to a particular order in a method claim or specification, no order is to be inferred in any respect. This applies to any possible non-express basis for interpretation, including matters of logic regarding the arrangement of steps or operational flows, ordinary meaning derived from grammatical organization or punctuation, or the number or type of aspects described in the specification.

[0140] All publications and patents cited in this specification are cited to disclose and describe the methods and / or materials in connection with the cited publications. All of these publications and patents are incorporated herein by reference as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. Such inclusion by reference is expressly limited to the methods and / or materials described in the cited publications and patents and does not extend to any dictionary definitions from the cited publications and patents. Any dictionary definitions in the cited publications and patents that are not explicitly repeated in this application should not be treated as such and should not be construed as defining any term appearing in the appended claims. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present disclosure is not entitled to antedate such publication by virtue of prior disclosure. In addition, the publication dates provided may be different from the actual publication dates, which may need to be independently confirmed.

[0141] The present disclosure describes novel compounds or pharmaceutically acceptable salts, enantiomers, stereoisomers or prodrugs thereof, which are represented by the following formula (I). In addition, the present disclosure describes analogs or free radical forms of the compounds represented by formula (I).

[0142]

[0143] In formula (I), in some embodiments, R 1is: C1-C8 alkyl, C3-C8 alkenyl, C3-C8 alkynyl, C3-C8 cycloalkyl or C5-C 10 Bicycloalkyl, each optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, NH2, C1-C3 haloalkyl, optionally substituted C6-C 10 Aryl, C1-C3 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C5-C 10 bicycloalkyl or optionally substituted C3-C6 heterocycloalkyl.

[0144] In some embodiments, X is: a bond; or -(CH2) n –, optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, C1-C3 haloalkyl, C3-C6 cycloalkyl or C1-C6 alkyl, wherein the C1-C6 alkyl group may form, together with the atoms to which it is attached, a C3-C6 spiroalkyl ring, and wherein –(CH2) n – The group may contain 0-1 double or triple bonds.

[0145] In some embodiments, n is an integer from 1 to 6.

[0146] In some embodiments, A is: a bond; C4-C8 cycloalkyl, C5-C 10 Bicycloalkyl, C5-C 10 Heterobicycloalkyl or C3-C8heterocycloalkyl, each optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, CN, C1-C3 haloalkyl, C1-C3 alkyl or C3-C5 cycloalkyl.

[0147] In some embodiments, Y is a bond or -(CH2) m –.

[0148] In some embodiments, m is an integer of 1 or 2.

[0149] In some embodiments, Z is H or halogen.

[0150] In some embodiments, R 2 Yes:-NR 12 R 5 、-NHC(O)R 6 、-NHC(O)NR 8 (R 9 )、-C(O)NHR 5 , five-membered or six-membered heteroaryl or C3-C8 heterocycloalkyl ring or C5-C 10 Heterobicycloalkyl ring, optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, CN, C1-C3 haloalkyl, C1-C3 alkyl, or C3-C5 cycloalkyl.

[0151] In some embodiments, R 5 is: H, C1-C5 alkyl, C3-C7 cycloalkyl, five-membered or six-membered heteroaryl, C5-C 10 Bicycloalkyl, (C3-C6 cycloalkyl)-C1-C3 alkyl, (C3-C6 heterocycloalkyl)-C1-C3 alkyl, (C6-C 10 (C1-C5-heteroaryl)-C1-C3 alkyl, or C3-C7 heterocycloalkyl, each optionally substituted with one or more halogen, CN, OH, O-(C1-C3 haloalkyl), O-(C1-C3 alkyl), C1-C3 haloalkyl, CH3SO2-, optionally substituted C3-C6 cycloalkyl, optionally substituted C3-C7 heterocycloalkyl, or C(O)NR 8 (R 9 ).

[0152] In some embodiments, R 6 is: C1-C6 alkyl, C3-C6 cycloalkyl or C3-C6 heterocycloalkyl, each optionally substituted with one or more halogen, C1-C3 haloalkyl, optionally substituted C3-C6 cycloalkyl or NR 8 (R 9 ).

[0153] In some embodiments, R 8 and R 9 are all independently H, substituted C1-C3 alkyl, or taken together with the nitrogen to which they are attached to form a 4-6 membered heterocyclic ring optionally substituted with one or more halogen or C1-C3 haloalkyl.

[0154] In some embodiments, R 12 is: H, C1-C5 alkyl, or the nitrogen to which it is attached and R 5 Together they form (i) a C3-C8 cycloalkyl ring or a heterocycloalkyl ring or (ii) a C5-C 10 A bicycloalkyl ring or a heterobicycloalkyl ring, each optionally substituted with one or more halogen, C1-C3 haloalkyl, OH, O-(C1-C3 alkyl), O-(C1-C3 haloalkyl) or CN.

[0155] In some embodiments, each stereocenter in the compound of Formula (I) is independently an R-enantiomer, an S-enantiomer, or a mixture of an R-enantiomer and an S-enantiomer.

[0156] In some embodiments, each double bond in the compound of Formula (I) is independently cis or trans.

[0157] In some embodiments, a compound or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof is provided, wherein the compound is represented by formula (Ia):

[0158]

[0159] In Formula (Ia), in some embodiments, X is a bond or -(CH2) n –.

[0160] In some embodiments, R 1 is: C1-C8 alkyl or C3-C8 cycloalkyl, each optionally substituted with one or more halogen, OH, NH2, C1-C3 alkyl, C1-C3 haloalkyl, O-(C1-C3 alkyl), optionally substituted C6 aryl, optionally substituted C3-C8 cycloalkyl, optionally substituted C3-C6 heterocycloalkyl or optionally substituted C6-C8 bicycloalkyl.

[0161] In some embodiments, A is: C4-C6 cycloalkyl, C4-C6 heterocycloalkyl, or C5-C7 bicycloalkyl, each optionally substituted with one or more halogen, OH, C1-C3 alkyl, or C1-C3 haloalkyl.

[0162] In some embodiments, Y is a bond or -CH2-.

[0163] In some embodiments, R 5 is: H, C1-C5 alkyl, C3-C6 cycloalkyl, 6-membered heteroaryl, C5 bicycloalkyl, (C3-C4 cycloalkyl)-C1-C2 alkyl, (C3-C4 heterocycloalkyl)-C1-C2 alkyl, (C6 aryl)-C1-C2 alkyl, (C5 heteroaryl)-C1-C2 alkyl or C3-C4 heterocycloalkyl, each optionally substituted with one or more halogen, CN, OH, O-(C1-C3 haloalkyl), O-(C1-C3 alkyl), C1-C3 haloalkyl, CH3SO2- or C(O)NR 8 (R 9 ).

[0164] In some embodiments, R 12 is H, C1-C5 alkyl, or the nitrogen to which it is attached and R 5 Together they form a C3-C8 heterocycloalkyl ring optionally substituted with one or more halogen or C1-C3 haloalkyl.

[0165] In some embodiments, a compound or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof is provided, wherein the compound is represented by formula (Ib):

[0166]

[0167] In Formula (Ib), in some embodiments, X is a bond or -(CH2) n –.

[0168] In some embodiments, R 1 is: C3-C6 alkyl, optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, NH2, C1-C3 haloalkyl.

[0169] In some embodiments, A is: a bond; or C4-C6 cycloalkyl, optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, NH2, C1-C3 haloalkyl.

[0170] In some embodiments, Y is a bond.

[0171] In some embodiments, R 6 is optionally replaced with NR 8 (R 9 ) of a C1-C6 alkyl group.

[0172] In some embodiments, a compound or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof is provided, wherein the compound is represented by formula (Ic):

[0173]

[0174] In Formula (Ic), in some embodiments, X is a bond or -(CH2) n –.

[0175] In some embodiments, R 1 is: C1-C6 alkyl, optionally substituted with optionally substituted C3-C8 cycloalkyl or optionally substituted C3-C6 heterocycloalkyl.

[0176] In some embodiments, A is: a bond; C4-C6 cycloalkyl, C4-C6 heterocycloalkyl, or C5-C7 bicycloalkyl, each optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, NH2, or C1-C3 haloalkyl.

[0177] In some embodiments, Y is a bond.

[0178] In some embodiments, R 5 is H or C1-C3 alkyl.

[0179] In some embodiments, a compound or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof is provided, wherein the compound is represented by formula (Id):

[0180]

[0181] In Formula (Id), in some embodiments, R 1 It is a C1-C3 alkyl group which is optionally substituted with an optionally substituted C3-C6 cycloalkyl group. For example, the C3-C6 cycloalkyl group may be substituted with one or more halogen, CN, OH, C1-C3 alkyl, O-(C1-C3 haloalkyl), O-(C1-C3 alkyl) or C1-C3 haloalkyl.

[0182] In certain embodiments, A is an optionally substituted C4-C6 cycloalkyl. For example, A may be substituted with one or more halogen, CN, OH, C1-C3 alkyl, O-(C1-C3 haloalkyl), O-(C1-C3 alkyl) or C1-C3 haloalkyl.

[0183] In some embodiments, R 2 is: C1-C4 alkyl or (C3-C4 cycloalkyl)-C1-C2 alkyl, each optionally substituted with one or more halogen, CN, OH, O-(C1-C3 haloalkyl), O-(C1-C3 alkyl), C1-C3 haloalkyl or CH3SO2-. For example, R 2 is C1-C3 alkyl optionally substituted with one or more halogen, O-(C1-C3 haloalkyl), O-(C1-C3 alkyl) or C1-C3 haloalkyl.

[0184] In Formula (I), in some embodiments, each X and Y is a bond; A is a C4-C6 cycloalkyl group; R 1 is C1-C3 alkyl optionally substituted with optionally substituted C3-C6 cycloalkyl; and R 2 It is a 5-membered heteroaryl group.

[0185] The present disclosure describes novel compounds represented by formula (I) below, or pharmaceutically acceptable salts, enantiomers, stereoisomers or prodrugs thereof. Additionally, the present disclosure describes analogs or free radical forms of the compounds represented by formula (I).

[0186] As used herein, unless otherwise indicated, the term "compound" refers to any specific chemical compound disclosed herein, and includes tautomers, positional isomers, geometric isomers, and applicable stereoisomers (including optical isomers (enantiomers) and other stereoisomers (diastereomers) thereof), as well as pharmaceutically acceptable salts and derivatives thereof (including prodrug forms) where applicable in the context. Within its use in this context, the term compound generally refers to a single compound, but may also include other compounds (such as stereoisomers, positional isomers and / or optical isomers (including racemic mixtures) and specific enantiomers or enantiomerically enriched mixtures of the disclosed compounds). In this context, the term also refers to a prodrug form of a compound, which has been modified to facilitate administration and delivery of the compound to the active site. It is noted that, in describing the present compounds, among other things, a number of substituents and variations associated therewith are described. It is understood by those of ordinary skill that the molecules described herein are stable compounds as generally described below.

[0187] It is understood that in any compound described herein with one or more chiral centers, if absolute stereochemistry is not clearly indicated, each center can independently be R-configuration or S-configuration or a mixture thereof. Therefore, the compound provided herein can be enantiomerically pure, enantiomerically enriched, or can be a stereoisomeric mixture, and includes all diastereoisomers and enantiomeric forms. In addition, it is understood that in any compound described herein with one or more double bonds that produce geometric isomers that can be defined as E or Z, each double bond can independently be E or Z or a mixture thereof. If desired, stereoisomers are obtained by methods such as stereoselective synthesis and / or by separating stereoisomers using a chiral chromatographic column. Similarly, it is understood that in any compound described, all tautomers and conformational isomers are also intended to be included. A conformational isomer is a structure as an isomer of conformation. Conformational isomerism is the phenomenon of molecules with the same structural formula but with different conformations (conformers) of atoms around a rotating bond.

[0188] As used herein, any "R" group represents a substituent that may be attached to the indicated atom. The R group may be substituted or unsubstituted. Whenever a group is described as "optionally substituted," the group may be unsubstituted or substituted with one or more of the indicated substituents. Similarly, when a group is described as "unsubstituted or substituted," if substituted, the substituents may be selected from one or more of the indicated substituents. If no substituents are indicated, it means that the indicated "optionally substituted" or "substituted" group may be individually and independently substituted with one or more groups individually and independently selected from alkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, aralkyl, heteroaralkyl, heterocyclyl(alkyl), hydroxy, alkoxy, cycloalkyloxy, aryloxy, acyl, mercapto, alkylthio, arylthio, cyano, halogen, C-amido, N-amido, C-carboxyl, O-carboxyl, isocyanato, thiocyanato, isothiocyanato, nitro, haloalkyl, haloalkoxy, amino (including monosubstituted and disubstituted amino), and alkylamino. When a group is not described as "optionally substituted," "unsubstituted," or "substituted," such group is unsubstituted unless a definition of such group is otherwise stated.

[0189] As used herein, where "a" and "b" are integers, a to C b ” or “C a -C b " refers to the number of carbon atoms in an alkyl, alkenyl, or alkynyl group, or the number of carbon atoms in a cycloalkyl, aryl, heteroaryl, heterocycloalkyl, or heterocyclyl ring. That is, the alkyl, alkenyl, alkynyl, cycloalkyl ring, aryl ring, heterocycloalkyl ring, heteroaryl ring, or heterocyclyl ring can contain from "a" to "b" (inclusive) carbon atoms.

[0190] As used herein, the term "alkyl" refers to a saturated monovalent chain of carbon atoms, which may be optionally branched. It is understood that in embodiments comprising an alkyl group, illustrative variations of those embodiments include lower alkyl groups (such as C1-C8 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, methyl, ethyl, propyl, 3-methylpentyl, etc.).

[0191] As used herein, the term "alkenyl" refers to a straight or branched hydrocarbon having at least 2 carbon atoms and at least one double bond. Alkenyl can include any number of carbons, such as C2, C2-C3, C2-C4, C2-C5, C2-C6, C2-C7, C2-C8, C3, C3-C4, C3-C5, C3-C6, C3-C7, C3-C8, C4, C4-C5, C4-C6, C4-C7, C4-C8, C5, C5-C6, C5-C7, C5-C8, C6, C6-C7, C6-C8, C7, C7-C8, and C8. Alkenyl can have any suitable number of double bonds, including but not limited to 1, 2, 3, 4, 5, or more. Examples of alkenyl groups include, but are not limited to, vinyl (vinyl), propenyl, isopropenyl, 1-butenyl, 2-butenyl, isobutenyl, butadienyl, 1-pentenyl, 2-pentenyl, isopentenyl, 1,3-pentadienyl, 1,4-pentadienyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 1,3-hexadienyl, 1,4-hexadienyl, 1,5-hexadienyl, 2,4-hexadienyl, or 1,3,5-hexatrienyl. The alkenyl group may be optionally substituted with one or more moieties selected from halogen, hydroxyl, amino, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxyde and cyano groups.

[0192] As used herein, the term "alkynyl" refers to a straight or branched chain hydrocarbon having at least 2 carbon atoms and at least one triple bond. Alkynyl can include any number of carbon atoms, such as C2, C2-C3, C2-C4, C2-C5, C2-C6, C2-C7, C2-C8, C3, C3-C4, C3-C5, C3-C6, C3-C7, C3-C8, C4, C4-C5, C4-C6, C4-C7, C4-C8, C5, C5-C6, C5-C7, C5-C8, C6, C6-C7, C6-C8, C7, C7-C8, and C8. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, 1-butynyl, 2-butynyl, isobutynyl, sec-butynyl, butadiynyl, 1-pentynyl, 2-pentynyl, isopentenyl, 1,3-pentadiynyl, 1,4-pentadiynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 1,3-hexadiynyl, 1,4-hexadiynyl, 1,5-hexadiynyl, 2,4-hexadiynyl, or 1,3,5-hexatriynyl. The alkynyl group may be optionally substituted with one or more moieties selected from halogen, hydroxyl, amino, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxyde and cyano groups.

[0193] As used herein, the term "haloalkyl" refers to an alkyl group in which one or more hydrogen atoms have been replaced by one or more halogen atoms. n-m Haloalkyl" or "C n-C m "Haloalkyl" refers to a C group having n to m carbon atoms and at least one to a maximum of {2(n to m) + 1} halogen atoms which may be the same or different. n-m Alkyl. In some embodiments, the halogen atom is a fluorine atom. In some embodiments, the haloalkyl group has 1 to 6 or 1 to 4 carbon atoms. Example haloalkyl groups include CF3, C2F5, CHF2, CH2F, CCl3, CHCl2, C2Cl5, etc. In some embodiments, the haloalkyl group is a fluoroalkyl group.

[0194] As used herein, the term "cycloalkyl" refers to a monovalent chain of carbon atoms a portion of which forms a ring. It is understood that in embodiments that include cycloalkyl, illustrative variations of those embodiments include lower cycloalkyl (such as C3-C8 cycloalkyl, C3-C7 cycloalkyl, C3-C6 cycloalkyl, C3-C5 cycloalkyl, C4-C8 cycloalkyl, C4-C7 cycloalkyl, C4-C6 cycloalkyl, cyclopropyl, cyclohexyl, 3-ethylcyclopentyl, etc.).

[0195] As used herein, the term "bicycloalkyl" refers to two cycloalkyl groups as defined above linked to each other to form a bridged, fused, or spiro bicyclic compound.

[0196] As used herein, the term "fused bicycloalkyl" refers to two cycloalkyl groups that share two adjacent atoms. In other words, the rings share one covalent bond, i.e., the so-called bridgehead atoms are directly connected.

[0197] As used herein, the term "bridged bicycloalkyl" refers to two cycloalkyl groups whose moieties have more than two atoms in common. That is, the two rings do not share adjacent carbon atoms, but rather three or more atoms, with the two bridgehead atoms separated by a bridge comprising at least one atom. Examples include, but are not limited to, bicyclo[3.2.1]heptyl ("norbornyl"), bicyclo[2.2.2]octyl, and the like.

[0198] As used herein, the term "spiro bicycloalkyl" refers to a bicyclic group wherein the two rings are connected at a single carbon atom as a member of each of the two rings. The term includes a spiro bicycloalkyl wherein the two rings are cycloalkyl rings connected at a single carbon atom as a member of each of the two rings and a spiro bicycloheteroalkyl wherein the two rings are connected at a single carbon atom as a member of each of the two rings and a ring wherein the two rings connected at a single carbon atom as a member of each of the two rings are heterocyclyl rings and the other ring is a cycloalkyl ring or a spiro bicycloheteroalkyl wherein the two rings connected at a single carbon atom as a member of each of the two rings are heterocyclyl rings. The example of spiro bicycloalkyl includes spiro [3.3] heptenyl, spiro [3.4] octanyl, azaspiro [3.3] heptanyl, oxazaspiro [3.3] heptanyl, oxazaspiro [3.3] heptanyl and azaspiro [3.4] octanyl.

[0199] As used herein, " heterocycloalkyl " refers to a non-aromatic ring with at least one heteroatom ring member independently selected from nitrogen, sulphur and oxygen. Heterocycloalkyl can include monocyclic or polycyclic (for example, fused, bridged or spiral) ring system. Heterocycloalkyl also includes one or more aromatic rings fused to non-aromatic heterocycloalkyl rings. The ring-forming carbon atoms of heterocycloalkyl can be replaced by oxygen substituents alternatively. The ring-forming heteroatoms of heterocycloalkyl can be oxidized to form N-oxide or sulfonyl. It is understood that in the embodiment comprising heterocycloalkyl, the illustrative variations of those embodiments include low cycloalkyl (such as C3-C8 heterocycloalkyl, C3-C7 heterocycloalkyl, C3-C6 heterocycloalkyl, C3-C5 heterocycloalkyl, C4-C8 heterocycloalkyl, C4-C7 heterocycloalkyl, C4-C6 heterocycloalkyl etc.). Examples of the heterocycloalkyl group include a morpholine ring, a pyrrolidine ring, a piperazine ring, a piperidine ring, a tetrahydropyran ring, a tetrahydropyridine, an azetidine ring, tetrahydrofuran and the like.

[0200] As used herein, "heterobicycloalkyl" refers to an unsubstituted or substituted bicycloalkyl structure in which at least one carbon atom is replaced with a heteroatom independently selected from oxygen, nitrogen, and sulfur.

[0201] As used herein, the term "aryl" refers to an aromatic carbocyclic ring system with any suitable number of ring atoms and any suitable number of rings. Aryl can include any suitable number of carbon ring atoms (such as 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 ring atoms) and 6 to 10, 6 to 12 or 6 to 14 ring members. Aryl can be monocyclic, fused to form a two-ring or three-ring group, or connected to form a biaryl group by a key. Representative aryl groups include phenyl, naphthyl and biphenyl. Other aryl groups include benzyl with a methylene linking group. Some aryl groups have 6 to 12 ring members (such as phenyl, naphthyl or biphenyl). Other aryl groups have 6 to 10 ring members (such as phenyl or naphthyl). Some other aryl groups have 6 ring members (such as phenyl). The aryl group may be optionally substituted with one or more moieties selected from alkyl, alkenyl, alkynyl, haloalkyl, halogen, hydroxy, amino, alkylamino, alkoxy, haloalkyl, carboxyl, alkylcarboxylate, amido, nitro, oxydimide, and cyano.

[0202] As used herein, the term "heteroaryl" refers to substituted and unsubstituted aromatic 5- or 6-membered monocyclic groups and 9- or 10-membered bicyclic groups having at least one heteroatom (O, S, or N) in at least one ring, the heteroatom-containing ring preferably having 1, 2, or 3 heteroatoms independently selected from O, S, and / or N. Each ring of the heteroaryl group containing heteroatoms can contain one or two oxygen atoms or sulfur atoms and / or one to four nitrogen atoms, provided that the total number of heteroatoms in each ring is four or less and each ring has at least one carbon atom. The fused rings constituting the bicyclic group are aromatic and can contain only carbon atoms. The nitrogen and sulfur atoms can be optionally oxidized, and the nitrogen atom can be optionally quaternized. The bicyclic heteroaryl group must only include aromatic rings. The heteroaryl group can be connected at any available nitrogen or carbon atom of any ring. The heteroaryl ring system can be unsubstituted or can contain one or more substituents. Examples of monocyclic heteroaryl groups include, but are not limited to, thiazolyl, oxazolyl, thienyl, furanyl, pyrrolyl, imidazolyl, isoxazolyl, pyrazolyl, triazolyl, thiadiazolyl, tetrazolyl, oxadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thiazolyl, and other similar groups. Examples of bicyclic heteroaryl groups include, but are not limited to, indolyl, benzothienyl, benzofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, quinolinyl, isoquinolinyl, purinyl, purinopyridinyl, oxochromenyl, dioxoisoindolinyl, pyrazolopyridinyl, pyrazolo[1,5-a]pyridinyl, and other similar groups.

[0203] In some embodiments, the compound is selected from Table A below.

[0204] [Table A]

[0205]

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221]

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232]

[0233]

[0234]

[0235]

[0236]

[0237]

[0238]

[0239]

[0240]

[0241]

[0242]

[0243]

[0244]

[0245]

[0246]

[0247]

[0248]

[0249]

[0250]

[0251]

[0252]

[0253]

[0254]

[0255]

[0256]

[0257]

[0258]

[0259]

[0260]

[0261]

[0262]

[0263]

[0264]

[0265]

[0266]

[0267]

[0268]

[0269]

[0270]

[0271]

[0272]

[0273]

[0274] In some embodiments, the compounds of the present disclosure are used to degrade and / or reduce casein kinase 1 alpha (CK1 alpha).

[0275] In some embodiments, the compounds of the present disclosure are used to inhibit casein kinase 1α (CK1α) activity.

[0276] In some embodiments, the compounds of the present disclosure are used to prevent or treat blood cancers.

[0277] In some embodiments, the present disclosure provides a pharmaceutical composition comprising: a compound of the present disclosure (represented by Formula (I), Formula (Ia), Formula (Ib) or Formula (Ic)), or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof; and optionally a pharmaceutically acceptable excipient or carrier.

[0278] The term "pharmaceutically acceptable" describes a material that is not biologically or otherwise undesirable, ie, does not induce unacceptable levels of undesired biological effects or interact in a deleterious manner.

[0279] The term "pharmaceutically acceptable carrier" is used herein to refer to a carrier useful for preparing a pharmaceutical composition that is generally safe, non-toxic, and neither biologically nor otherwise undesirable, and is acceptable for veterinary use as well as human pharmaceutical use. As used in the specification and claims, "pharmaceutically acceptable carrier" can include one or more than one such carrier. "Pharmaceutically acceptable" means that the carrier must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

[0280] As used herein, the term "pharmaceutically acceptable salt" means a salt of an active ingredient prepared with an acid or base, which, when administered in a therapeutically effective amount, is tolerated by the biological system or tolerated by the subject or tolerated by the biological system and tolerated by the subject. When the compounds of the present disclosure contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compound with a sufficient amount of the desired base (neat or in a suitable inert solvent). Examples of pharmaceutically acceptable base addition salts include, but are not limited to, sodium salts, potassium salts, calcium salts, ammonium salts, organic amine salts, magnesium salts, lithium salts, strontium salts, or similar salts. When the compounds of the present disclosure contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compound with a sufficient amount of the desired acid (neat or in a suitable inert solvent). Examples of pharmaceutically acceptable acid addition salts include, but are not limited to, salts derived from inorganic acids (e.g., hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid, or phosphorous acid, and the like) and salts derived from relatively non-toxic organic acids (e.g., acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and the like). Also included are salts of amino acids (e.g., arginine, and the like) and salts of organic acids (e.g., glucuronic acid or galacturonic acid, and the like).

[0281] The compound represented by formula (I) or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof degrades casein kinase 1α (CK1α).

[0282] In some embodiments, the pharmaceutical composition is used to degrade and / or reduce casein kinase 1 alpha (CK1 alpha).

[0283] In some embodiments, the pharmaceutical composition is used to inhibit casein kinase 1α (CK1α) activity.

[0284] In some embodiments, the pharmaceutical composition is used to prevent or treat blood cancer.

[0285] In some embodiments, the blood cancer is leukemia, lymphoma, or myeloma.

[0286] The present disclosure also provides a method of degrading and / or reducing casein kinase 1α (CK1α) in a subject in need thereof, the method comprising administering to the subject an effective amount of the pharmaceutical composition of the present disclosure.

[0287] The present disclosure also provides a method of inhibiting casein kinase 1α (CK1α) activity in a subject in need thereof, the method comprising administering to the subject an effective amount of the pharmaceutical composition of the present disclosure.

[0288] The present disclosure also provides a method for preventing or treating blood cancer in a subject in need thereof, comprising administering to the subject an effective amount of the pharmaceutical composition of the present disclosure.

[0289] As used herein, the term "blood cancer" refers to cancer that begins in cells of hematopoietic tissue (such as the bone marrow) or the immune system. In some embodiments, "blood cancer" can include any cancer that involves the uncontrolled proliferation of blood cells (particularly, white blood cells).

[0290] In some embodiments, the blood cancer is leukemia, lymphoma (Hodgkin's lymphoma and non-Hodgkin's lymphoma), or myeloma.

[0291] In some embodiments, the leukemia is selected from the group consisting of acute myeloid leukemia (AML), acute lymphoblastic leukemia or acute lymphoblastic leukemia (ALL), T-cell acute lymphoblastic leukemia (T-ALL), B-cell acute lymphoblastic leukemia (B-ALL), chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), T-cell prolymphocytic leukemia (T-PLL), large granular lymphocytic leukemia, adult T-cell leukemia, chronic eosinophilic leukemia (CEL) and myelodysplastic syndrome (MDS).

[0292] In some embodiments, the lymphoma is selected from the group consisting of Hodgkin lymphoma, diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, small lymphocytic lymphoma (SLL), mantle cell lymphoma (MCL), marginal zone lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, primary central nervous system (CNS) lymphoma, and peripheral T-cell lymphoma.

[0293] In some embodiments, the myeloma is selected from the group consisting of multiple myeloma, light chain myeloma, non-secretory myeloma, solitary plasmacytoma, extramedullary plasmacytoma, monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM), immunoglobulin D (IgD) myeloma, and immunoglobulin E (IgE) myeloma.

[0294] In some embodiments, compounds of the present disclosure can bind to cereblon (CRBN), altering the specificity of the complex to induce ubiquitination and degradation of CK1α, a transcription factor essential for the growth of proliferative diseases such as multiple myeloma.

[0295] In some embodiments, the present disclosure provides compounds and compositions comprising an E3 ligase binding portion that can bind to an E3 ligase (e.g., CRBN) and a target protein binding portion that can bind to a target protein, resulting in ubiquitination of the target protein and degradation of the target protein by the proteasome.

[0296] As used herein, the term "degradation" or variations thereof as used herein means degradation of a target protein mediated by an E3 ligase (e.g., CRBN), resulting in a decrease in protein levels. In one embodiment, the target protein may be a casein kinase 1 (CK1, e.g., CK1α, CK1β1, CK1γ1, CK1γ2, CK1γ3, CK1δ, CK1ε) protein. In certain embodiments, the target protein may preferably be CK1α.

[0297] As used herein, the term "CK1α" refers to casein kinase 1α, a kinase encoded by the CSNK1A1 gene in humans. CK1α has been shown to play a key role in the biology of AML (Jaras M et al, J Exp Med. 2014; 211(4): 605-612).

[0298] As used herein, the term "effective amount" refers to the amount of an agent administered that is sufficient to degrade CK1α and / or inhibit CK1α activity. In certain embodiments, the term "effective amount" refers to the amount of an agent administered that is sufficient to treat a CK1α-related disease (preferably, a blood cancer).

[0299] As used herein, the term "treat" or variations thereof refers to administering a compound or pharmaceutical composition to a subject for prophylactic and / or therapeutic purposes. The term "prophylactic treatment" refers to treating a subject who does not yet exhibit symptoms of a disease or condition, but is susceptible to or otherwise at risk for a particular disease or condition, thereby reducing the likelihood that the patient will develop the disease or condition. The term "therapeutic treatment" refers to administering treatment to a subject who already has a disease or condition.

[0300] As used herein, the term "prevent" refers to slowing down a disease or slowing down the onset of a disease or its symptoms. Preventing a disease or condition can include stopping the onset of the disease or its symptoms.

[0301] The pharmaceutical compositions described herein can be administered to human patients by themselves, or in the form of pharmaceutical compositions (wherein they are mixed with other active ingredients or excipients, or combinations thereof, as in combination therapy). The appropriate formulation depends on the chosen route of administration. The techniques for the preparation and administration of the compounds described herein are known to those skilled in the art.

[0302] The pharmaceutical compositions disclosed herein can be prepared in a manner known per se (e.g., by conventional mixing, dissolving, granulating, making dragees, grinding, emulsifying, encapsulating, embedding, or tableting processes). Additionally, the active ingredient is included in an amount effective to achieve its intended purpose. Many of the compounds used in the pharmaceutical combinations disclosed herein can be provided as salts with pharmaceutically compatible counterions.

[0303] There are a variety of techniques for administering compounds, salts and / or compositions in the art, including but not limited to oral delivery, rectal delivery, pulmonary delivery, local delivery, aerosol delivery, injection delivery, infusion delivery and parenteral delivery, including intramuscular injection, subcutaneous injection, intravenous injection, intramedullary injection, intrathecal injection, direct intraventricular injection, intraperitoneal injection, intranasal injection and intraocular injection. In some embodiments, the compounds described herein (including compounds of formula (I)) or pharmaceutically acceptable salts, enantiomers, stereoisomers or prodrugs thereof can be administered orally.

[0304] Compounds, salts and / or compositions can also be administered in a local rather than systemic manner, for example, by direct injection or implantation of the compound into the affected area, typically in the form of a reservoir or sustained-release formulation. In addition, the compound can be administered in a targeted drug delivery system (e.g., in a liposome coated with a tissue-specific antibody). The liposomes will target organs and be selectively absorbed by the organs. For example, intranasal delivery or pulmonary delivery of targeted respiratory diseases or conditions can be desirable.

[0305] If desired, the composition can be present in a package or dispensing device, which can include one or more unit dosage forms containing an active ingredient. Package can, for example, include metal or plastic foil (such as a blister pack). Package or dispensing device can be provided with instructions for administration. Package or dispensing device can also be provided with precautions associated with the container, which are in the form prescribed by a government agency that manages the production, use, or sale of the drug, and which reflect that the drug form has been approved by the agency for human or veterinary administration. Such precautions, for example, can be labels for prescription drugs approved by the U.S. Food and Drug Administration or product instructions for approval. Compositions comprising compounds described herein and / or salts can also be prepared in suitable containers, placed in compatible pharmaceutical excipients, and marked for the treatment of specified diseases.

[0306] Compound, salt and / or pharmaceutical composition can be provided to the administering physician or other healthcare professional in the form of a kit. Kit is a container containing the compound in a suitable pharmaceutical composition and a package containing instructions for administering the pharmaceutical composition to the subject. Kit may also optionally include one or more additional therapeutic agents. Kit may also include a single dose of compound or pharmaceutical composition for continuous or sequential administration. Kit may optionally include one or more diagnostic tools and instructions for use. Kit may include a suitable delivery device (e.g., syringe, etc.) and instructions for administering the compound and any other therapeutic agent. Kit may optionally include instructions for storing, reconstructing (if applicable) and administering any or all therapeutic agents included. Kit may include multiple containers reflecting the number of administrations given to the subject.

[0307] As will be apparent to those skilled in the art, useful in vivo dosages and specific modes of administration will vary according to age, body weight, severity of suffering, and species of mammal being treated, the specific compounds used, and the specific uses for which these compounds are used. The determination of effective dosage levels (i.e., dosage levels necessary to achieve the desired result) can be accomplished by those skilled in the art using conventional methods (e.g., human clinical trials and in vitro studies).

[0308] Dosage can be widely changed according to desired effect and treatment indication.Alternatively, as will be appreciated by those skilled in the art, dosage can be based on the surface area of ​​the patient and calculated according to the surface area of ​​the patient.Although the exact dosage will be determined based on drug analysis one by one, in most cases, some generalizations can be made about dosage.The daily dosing regimen of adult patients can be, for example, an oral dose between about 0.01mg and 3,000mg of each active ingredient (preferably, between about 1mg and 700mg, for example, about 5 to 200mg).According to the needs of the subject, dosage can be a single dose or a series of two or more doses given in the process of one or more days.In certain embodiments, the compound is administered during continuous treatment, for example, one week or several weeks, or several months or several years.

[0309] In cases where human dosages have been established for compounds for at least some conditions, those same dosages or dosages between about 0.1% and 500% (more preferably, between about 25% and 250%) of established human dosages may be used. In cases where human dosages have not been established, as in the case of newly discovered pharmaceutical compositions, the ED may be used. 50 or Id 50 Suitable human doses can be extrapolated from values ​​obtained from in vitro studies or other appropriate values ​​derived from in vivo studies (e.g., as determined by toxicity studies and efficacy studies in animals).

[0310] In the case of administering a pharmaceutically acceptable salt, the dosage can be calculated as the free base. As will be appreciated by those skilled in the art, in certain circumstances, it may be necessary to administer a compound disclosed herein in an amount exceeding or even far exceeding the above-mentioned preferred dosage range to effectively and rapidly treat a rapidly developing disease or infection.

[0311] The dosage and interval can be adjusted individually to provide a plasma level of the active moiety sufficient to maintain the modulatory effect or minimum effective concentration (MEC). The MEC will vary for each compound, but can be estimated by in vitro data. The dosage required to reach the MEC will depend on individual characteristics and route of administration. However, HPLC assays or bioassays can be used to determine plasma concentrations. Dosage intervals can also be determined using MEC values. The composition should be administered using a regimen that maintains plasma levels above the MEC for about 10 to 90% (preferably, about 30 to 90%, most preferably about 50 to 90%) of the time. In the case of local administration or selective absorption, the effective local concentration of the drug may be independent of the plasma concentration.

[0312] It should be noted that due to toxicity or organ dysfunction, the attending physician will know how and when to terminate, interrupt or regulate administration. On the contrary, if the clinical response is insufficient (excluding toxicity), the attending physician will also know to adjust the treatment to a higher level. The size of the dosage in the treatment of the illness of interest will vary with the severity of the illness to be treated and the route of administration. The severity of the illness can, for example, be assessed in part by standard prognostic assessment methods. In addition, dosage and possible dosage frequency will also vary according to age, body weight and reaction of individual patients. A scheme suitable with the scheme discussed above can be used in veterinary medicine.

[0313] The effectiveness and toxicity of the compounds disclosed herein can be assessed using known methods. For example, the toxicity of a specific compound or a subset of the compounds sharing certain chemical moieties can be established by measuring the toxicity of a cell line (such as a mammalian cell line, preferably a human cell line) in vitro. The results of such studies typically predict toxicity in animals (such as mammals, or more specifically, humans). Alternatively, the toxicity of a specific compound in an animal model (such as a mouse, rat, rabbit, or monkey) can be determined using known methods. The effectiveness of a specific compound can be determined using several generally recognized methods (such as in vitro methods, animal models, or human clinical trials). When selecting a model to determine effectiveness, the technician can select a suitable model, dosage, route of administration, and / or regimen under the guidance of prior art.

[0314] Example

[0315] I. Synthesis of Compounds

[0316] Hydrochloride form of compound 1

[0317] 3-(4-((4-Aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0318]

[0319] Step 1: tert-Butyl (4-oxobutyl)carbamate

[0320]

[0321] Under nitrogen atmosphere, at -78 DEG C, over 15 minutes, a solution of oxalyl chloride (5.4mL, 63.49mmol, 1.2 equivalents) in DCM (50mL) was added dropwise to a solution of DMSO (10mL, 1 volume) in DCM (100mL, 10 volumes), and stirred at the same temperature for 15 minutes. Over 15 minutes, a solution of tert-butyl (4-hydroxybutyl) carbamate 1 (10g, 52.91mmol, 1 equivalent) in DCM (50mL) was added dropwise. After the reaction mixture was stirred at -78 DEG C for 30 minutes, N, N-diisopropylethylamine (29.2mL, 158.7mmol, 3 equivalents) was added dropwise at -78 DEG C. Once the addition was complete, the reaction mixture was slowly warmed to 0 DEG C and stirred for 30 minutes. The reaction was completed (by TLC analysis (2:8 / EtOAc: petroleum ether, R f ~0.6, KMnO4) confirmed), the reaction mixture was diluted with DCM (200 mL) and washed with 10% aqueous citric acid (1×100 mL), water (1×200 mL) and brine (1×200 mL), dried over Na2SO4, filtered and concentrated under vacuum below 30°C to afford tert-butyl (4-oxobutyl)carbamate (10 g, crude product) as a light yellow liquid, which was used further without purification.

[0322] 1 H-NMR (400MHz, DMSO-d6): δ9.66(s,1H),6.84(s,1H),2.89-2.93(m,2H),2.42-2.44(m,2H),1.60-1.64(m,2H),1.38(s,9H).

[0323] Step 2: tert-Butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate

[0324]

[0325] Under a nitrogen atmosphere, a mixture of tert-butyl (4-oxobutyl) carbamate 2 (9.0 g, 0.0484 mol, 1.8 equivalents) and lenalidomide 3 (CAS#191732-72-6, 7.0 g, 0.0269 mol, 1 equivalent) was dissolved in 1,2-dichloroethane (70 mL, 10 volumes) and DMF (70 mL, 10 volumes). Acetic acid (4.8 mL, 0.0807 mol, 3.0 equivalents) was added thereto at room temperature. The resulting reaction mixture was stirred at room temperature for 2 hours. At this stage, NaCNBH (6.6 g, 0.1076 mol, 4 equivalents) was added in batches at 0 ° C, and stirred at room temperature for 16 hours. The reaction was completed (by TLC analysis (8: 2 / EtOAc: petroleum ether, R f After 4 hours of heating (1 hour ℃ of 5min), the reaction mixture was diluted with DCM (200 mL) and washed with water (3×200 mL) and brine (200 mL), dried over Na 2 SO 4 , filtered and concentrated. The resulting product was triturated with hexanes (50 mL), filtered and suction dried to afford tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 4 (8 g, 69%) as a white solid.

[0326] 1 H-NMR (400MHz, DMSO-d6): δ11.08(s,1H),7.28(t,J=7.60Hz,1H),6.93(d,J=7.20Hz,1H),6.8 2(t,J=5.60Hz,1H),6.75(d,J=8.00Hz,1H),5.58(t,J=5.60Hz,1H),5.10-5.14(m,1H),4.10-4 .25(m,2H),3.11-3.14(m,2H),2.90-2.98(m,3H),2.60-2.64(m,1H),2.28-2.32(m,1H),2.04-2.06(m,1H),1.46-1.58(m,4H),1.37(s,9H).LC-MS:429.2(MH),Rt(min):2.552,Area%-97.64.

[0327] Step 3: 3-(4-((4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0328]

[0329] To an ice-cold suspension of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 4 (800 mg, 1.86 mmol, 1 eq) in DCM (10 mL) was added HCl in EtOAc (1 M, 8 mL) and stirred at room temperature for 4 hours. The reaction progress was monitored by LCMS. The reaction mixture was concentrated and lyophilized to afford 3-(4-((4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (530 mg, 79%) as a yellow solid.

[0330] 1 H-NMR (400MHz, DMSO-d6): δ11.02(s,1H),8.07(s,3H),7.34(t,J=7.60Hz, 1H),7.05(d,J=7.60Hz,1H),6.92(d,J=8.00Hz,1H),5.10-5.15(m,1H),4.1 9-4.37(m,2H),3.19-3.20(m,2H),2.90-2.95(m,1H),2.80-2.89(m,2H),2 .51-2.52(m,1H),2.29-2.33(m,1H),2.03-2.06(m,1H),1.66-1.67(m,4H). 13 C-NMR (100 MHz, DMSO-d6): δ 173.34, 171.52, 168.67, 140.34, 133.14, 129.87, 117.06, 114.76, 52.08, 46.58, 44.46, 31.65, 24.90, 23.23. LCMS: 331.3 (M+H). Method: Mobile phase A: 0.1% formic acid in H2O. Mobile phase B: acetonitrile. Flow rate: 1.5 mL / min. Column: Atlantis dC18 (50 × 4.6) mm, 5 μm. Rt (min): 1.638; Area % -99.32. HPLC method: Mobile phase A: 0.1% TFA in H2O. Mobile phase B: acetonitrile. Flow rate: 1.0 mL / min. Column: Atlantis dC18 (250×4.6) mm, 5 μm. Rt (min): 7.403; Area %: 98.88.

[0331] Hydrochloride form of compound 2

[0332] 3-(4-((4-Aminobutyl)(methyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0333]

[0334] Step 4: tert-Butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(methyl)amino)butyl)carbamate

[0335]

[0336] Under a nitrogen atmosphere, tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 4 (2 g, 4.65 mmol, 1 eq) and paraformaldehyde (1.39 g, 46.5 mmol, 10 eq) were added together to a mixture of 1,2-dichloroethane (20 mL, 10 volumes) and DMF (10 mL, 5 volumes). Acetic acid (0.8 mL, 13.95 mmol, 3 eq) was added thereto, and the reaction mixture was stirred at room temperature for 1 hour. NaCNBH (1.15 g, 18.6 mmol, 4 eq) was then added portionwise, and the reaction mixture was stirred at room temperature for another 16 hours. The reaction was complete (by TLC analysis (100% EtOAc R f The mixture was stirred for 2 hours at room temperature for 10 minutes (10 min, 37 min) and then stirred for 2 hours at room temperature for 3 hours (10 min, 37 min) (4 min, 38 min, 8 min), and 4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(methyl)amino)butyl)carbamate (800 mg, 39%) as a white solid.

[0337] 1 H-NMR (400MHz, DMSO-d6): δ11.02(s,1H),7.37(t,J=7.60Hz,1H),7.15(d,J=7.20 Hz,1H),6.99(d,J=8.00Hz,1H),6.80(t,J=5.60Hz,1H),5.08-5.13(m,1H),4.33- 4.51(m,2H),2.90-2.96(m,3H),2.88(s,3H),2.52-2.62(m,2H),2.01-2.03(m,1H ), 1.42-1.48 (m, 4H), 1.40 (s, 9H). LC-MS: 445.4 (MH), Rt (min): 2.525, area % -99.11.

[0338] Step 5: 3-(4-((4-aminobutyl)(methyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0339]

[0340] To an ice-cold solution of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(methyl)amino)butyl)carbamate 5 (800 mg, 1.80 mmol) in DCM (10 mL) was added HCl in EtOAc (1 M, 8 mL) and stirred at room temperature for 4 hours. The reaction progress was monitored by LCMS. The reaction mixture was concentrated and lyophilized to afford 3-(4-((4-aminobutyl)(methyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (550 mg, 88%) as a light yellow solid.

[0341] 1 H-NMR (400MHz, DMSO-d6): δ11.01(s,1H),7.99(brs,3H),7.37-7.51(m,3H),5.10-5.15(m,1H),4.44-4.63(m,2H),3.3 0(s,2H),3.01(s,3H),2.92-2.98(m,2H),2.60-2.78(m,3H),2.45-2.50(m,1H),2.06-2.08(m,1H),1.66-1.67(m,4H). 13 C-NMR (100 MHz, DMSO-d6): δ 173.34, 171.30, 167.67, 134.36, 133.03, 130.23, 117.06, 122.84, 55.43, 52.21, 48.02, 42.50, 31.65, 23.02, 22.96. LCMS: 345.1 (M+H). Method: Mobile phase A: 0.1% formic acid in H2O. Mobile phase B: acetonitrile. Flow rate: 1.5 mL / min. Column: Atlantis dC18 (50 × 4.6) mm, 5 μm. Rt (min): 1.252; Area % -98.76. HPLC method: Mobile phase A: 0.1% TFA in H2O. Mobile phase B: acetonitrile. Flow rate: 1.0 mL / min. Column: Atlantis dC18 (250×4.6) mm, 5 μm. Rt (min): 6.313; Area %: 97.81.

[0342] Hydrochloride form of compound 3

[0343] 3-(4-((4-Aminobutyl)(isopropyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0344]

[0345] Step 6: tert-Butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(isopropyl)amino)butyl)carbamate

[0346]

[0347] tert-Butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 4 (1.5 g, 3.4 mmol, 1 eq) and 2-methoxypropylene (1.25 g, 17.5 mmol, 5 eq) were added together in 1,2-dichloroethane (30 mL, 20 volumes) and the suspension was cooled to 0 ° C. Trifluoroacetic acid (0.22 mL, 3.0 mmol, 0.9 eq) was added dropwise thereto. Sodium triacetoxyborohydride (1.07 g, 5.1 mmol, 1.5 eq) was then added portionwise at 0 ° C under a nitrogen atmosphere and the resulting reaction mixture was stirred at room temperature for 16 hours. The reaction was complete (by TLC analysis (100% EtOAc R f After 4 hours of heating (10 minutes, 1 hour, 1 hour, 3 hours of heating), the reaction mixture was diluted with DCM (100 mL) and washed with water (2 x 100 mL) and brine (100 mL), dried over NaSO, filtered and concentrated under vacuum. The crude product was purified by chromatography using 40-45% ethyl acetate in dichloromethane (column size: Biotage R snap cartridge, KP-Sil, 50 g, 230-400 silica gel) to afford tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(isopropyl)amino)butyl)carbamate (620 mg, 38%) as a white solid.

[0348] LCMS: 473.4 (M+H). Method: Column: Atlantis dC18 (50×4.6) mm 5 μm,

[0349] Mobile phase: A: 0.1% formic acid in H2O; B: ACN; Flow rate: 1.5 mL / min; Rt (min): 2.454; Area % -97.90.

[0350] Step 7: 3-(4-((4-aminobutyl(isopropyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0351]

[0352] To an ice-cold solution of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(isopropyl)amino)butyl)carbamate 6 (620 mg, 1.313 mmol) in DCM (4 mL) was added HCl in EtOAc (1 M, 4 mL) and stirred at room temperature for 12 hours. The reaction progress was monitored by LCMS. The reaction mixture was concentrated under vacuum and further lyophilized to afford the desired compound 3-(4-((4-aminobutyl(isopropyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (440 mg, 96%) as a light brown solid.

[0353] 1 H-NMR (400MHz, DMSO-d6): δ11.05(s,1H),7.98(s,3H),7.72(s,2H),5.12-5.08(m,1H),4.56(dd,J=18.00,56.40Hz,2H),3.82-3.51(m,1H),3.47(s ,2H),2.90-2.70(m,1H),2.68-2.65(m,3H),2.44-2.41(m,1H),2.09(d,J= 5.20Hz, 1H), 1.53 (t, J = 7.60Hz, 2H), 1.33 (d, J = 6.80Hz, 2H), 1.23 (s, 6H). 13 C-NMR (100 MHz, DMSO-d6): 173.37, 171.25, 167.38, 136.43, 134.63, 130.36, 126.85, 123.25, 60.12, 52.33, 49.80, 48.17, 40.49, 39.24, 38.53, 31.55, 24.53, 22.91, 18.92, 18.62. LCMS: 373.2 (M-HCl). Method: Column: Atlantis dC18 (50×4.6) mm 5 μm,

[0354] Mobile phase: A: 0.1% formic acid in H2O; B: ACN; Flow rate: 1.5 mL / min. Rt (min): 1.239; Area % -97.330. HPLC: 97.905%, Rt (min): 6.141.

[0355] Hydrochloride form of compound 4

[0356] 3-(4-((4-Aminobutyl)(propyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0357]

[0358] Step 8: tert-Butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(propyl)amino)butyl)carbamate

[0359]

[0360] Under a nitrogen atmosphere, tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 4 (1.5 g, 3.4 mmol, 1 eq) and propionaldehyde (2.0 g, 3.4 mmol, 10 eq) were dissolved in a mixture of 1,2-dichloroethane (20 mL) and DMF (10 mL). Acetic acid (0.58 mL, 10.2 mmol, 3 eq) was added thereto, and the resulting reaction mixture was stirred at room temperature for 1 hour. At this point, sodium cyanoborohydride (0.84 g, 1.3 mmol, 4 eq) was added portionwise, and stirred at room temperature for 16 hours. The reaction was complete (by TLC analysis (100% EtOAc R f The mixture was stirred at 4 ℃ for 1 hr at 4 ℃ for 2 h (10 min, 3 saturates / min).Then the mixture was stirred for 2 hours at 4 ℃ for 30 min (1 ...

[0361] LCMS: 473.4 (M+H). Method: Column: Atlantis dC18 (50×4.6) mm 5 μm,

[0362] Mobile phase: A: 0.1% formic acid in H2O; B: ACN; Flow rate: 1.5 mL / min; Rt (min): 2.709; Area % -99.720.

[0363] Step 9: 3-(4-((4-aminobutyl)(propyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0364]

[0365] To an ice-cold solution of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(propyl)amino)butyl)carbamate 7 (600 mg, 1.2695 mmol, 1 eq) in DCM (10 mL) was added HCl in EtOAc (1 M, 5 mL) and stirred at room temperature for 4 hours. The reaction progress was monitored by LCMS. The reaction mixture was concentrated and further lyophilized to afford 3-(4-((4-aminobutyl)(propyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (360 mg, 68%) as a light yellow solid.

[0366] 1 H-NMR (400MHz, DMSO-d6): δ11.01(s,1H),7.99(s,3H),7.53(s,2H),5.12( q,J=4.80Hz,1H),4.49(d,J=36.40Hz,2H),3.29(s,1H),2.97(s,1H),2.96- 2.88(m,1H),2.72(q,J=22.80Hz,2H),2.62(d,J=16.40Hz,1H),2.44(s,1H) ,2.06(t,J=5.20Hz,1H),2.05(s,1H),1.55(s,6H),0.84(t,J=7.20Hz,3H). 13 C-NMR (100 MHz, DMSO-d6): δ 173.34, 171.27, 167.49, 162.82, 134.55, 130.45, 125.14, 57.35, 55.02, 52.34, 48.12, 38.58, 36.28, 31.59, 31.26, 24.51, 22.94, 21.57, 19.04, 11.25. LCMS: 373.1 (M-HCl). Method: Column: Atlantis dC18 (50×4.6) mm 5 μm, Mobile Phase: A: 0.1% formic acid in H2O, B: ACN, Flow Rate: 1.5 mL / min. Rt (min): 1.386; Area %: -98.551. HPLC: 98.878%, Rt(min): 6.95.

[0367] The hydrochloride form of compound 6

[0368] 3-(4-((2-(1-(aminomethyl)cyclohexyl)ethyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0369]

[0370] Step 1: 2-(1-(((tert-Butoxycarbonyl)amino)methyl)cyclohexyl)acetic acid

[0371]

[0372] To a solution of 2-(1-(aminomethyl)cyclohexyl)acetic acid 1 (30 g, 175.4 mmol, 1 eq) in THF (300 mL, 10 vol) was added dropwise a solution of NaOH (7.0 g, 175.4 mmol, 1 eq) in water (100 mL, 5 vol) and Boc-anhydride (38.2 g, 175.4 mmol, 1 eq) at 0°C. The mixture was stirred at room temperature for 16 hours. The reaction was complete (by TLC analysis (20% EtOAc in petroleum ether, R f To the 4-thiazolinyl group (100mL) of ...

[0373] LCMS (ELSD): 270.3 (MH). Method: Column: Atlantis dC18 (50×4.6) mm 5 μm,

[0374] Mobile phase: A: 0.1% formic acid in H2O; B: ACN; Flow rate: 1.5 mL / min; Rt (min): 2.950; Area % -99.502.

[0375] Step 2: Methyl 2-(1-(((tert-Butoxycarbonyl)amino)methyl)cyclohexyl)acetate

[0376]

[0377] To a solution of 2-(1-(((tert-butoxycarbonyl)amino)methyl)cyclohexyl)acetic acid 2 (20 g, 73.8 mmol, 1 eq) in DMF (200 mL, 10 volumes) was added potassium carbonate (20.3 g, 147.6 mmol, 2 eq) and iodomethane (26 mL, 369 mmol, 5 eq) dropwise at 0°C. The mixture was stirred at room temperature for 16 hours. The reaction was complete (by TLC analysis (20% EtOAc in petroleum ether, R fAfter 4 hours of heating (1 hour after 4 hours of heating), the reaction mixture was diluted with ethyl acetate (200 mL) and washed with ice-cold H2O (3×500 mL). The organic layer was washed with brine (1×500 mL), dried over Na2SO4, filtered and concentrated under vacuum to afford methyl 2-(1-(((tert-butoxycarbonyl)amino)methyl)cyclohexyl)acetate 3 (15 g, 71%) as a light yellow liquid.

[0378] LCMS (ELSD): 230.3 (M-tert-butyl). Method: Column: Atlantis dC18 (50×4.6) mm 5 μm,

[0379] Mobile phase: A: 0.1% formic acid in H2O; B: ACN; Flow rate: 1.5 mL / min; Rt (min): 3.374; Area % -99.76.

[0380] Step 3: tert-Butyl 3-hydroxy-2-azaspiro[4.5]decane-2-carboxylate

[0381]

[0382] To a solution of methyl 2-(1-(((tert-butoxycarbonyl)amino)methyl)cyclohexyl)acetate 3 (8 g, 28.07 mmol, 1 eq) in DCM (150 mL) was added DIBAL-H (1 M in hexanes, 28.07 mL, 28.07 mmol, 1 eq) dropwise at -78°C. The mixture was stirred at 0°C for 2 h.

[0383] The reaction was complete (by TLC analysis (20% EtOAc in petroleum ether, R f ~0.5) and LCMS-ELSD confirmed), the reaction mixture was quenched with saturated NH4Cl solution (50 mL) and filtered through a celite bed and washed with DCM (250 mL). The separated organic layer was washed with brine (250 mL), dried over Na2SO4, filtered and concentrated under vacuum below 35°C to afford tert-butyl 3-hydroxy-2-azaspiro[4.5]decane-2-carboxylate 4 (6 g, crude product) as a light brown liquid, which was used without purification.

[0384] LCMS (ELSD): 202.1 (M-tert-butyl). Method: Column: Atlantis dC18 (50×4.6) mm 5 μm,

[0385] Mobile phase: A: 0.1% formic acid in H2O; B: ACN; Flow rate: 1.5 mL / min; Rt (min): 2.187; Area % -99.976.

[0386] Step 4: tert-Butyl ((1-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethyl)cyclohexyl)methyl)carbamate

[0387]

[0388] Under a nitrogen atmosphere, acetic acid (1.82 mL, 30.7 mmol, 4 equivalents) was added to a stirred solution of lenalidomide (CAS#191732-72-6, 2 g, 7.6 mmol, 1 equivalent) and 3-hydroxy-2-azaspiro [4.5] decane-2-carboxylic acid tert-butyl ester 4 (5.79 g, 2.3 mmol) in a mixture of 1,2-dichloroethane (20 mL) and DMF (20 mL). The resulting reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was then cooled to 0 ° C, and sodium cyanoborohydride (1.84 g, 30.7 mmol, 4 equivalents) was added portionwise and stirred at room temperature for 16 hours. The reaction was completed (by TLC analysis (100% EtOAc, R f After 4 hours of heating (1 hour after 4 hours of heating), the reaction mixture was diluted with DCM (100 mL), washed with water (3×50 mL) and brine (100 mL), dried over Na 2 SO 4 , filtered and concentrated. The resulting residue was purified by chromatography using 40-45% EtOAc in DCM (column size: Biotage R snap cartridge, KP-Sil, 25 g, 230-400 silica gel) to afford tert-butyl ((1-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethyl)cyclohexyl)methyl)carbamate 5 (620 mg, 16%) as a white solid.

[0389] LCMS: 443.4 (Mt-Bu). Method: Column: Atlantis dC18 (50×4.6) mm 5 μm,

[0390] Mobile phase: A: 0.1% formic acid in H2O; B: ACN; Flow rate: 1.5 mL / min; Rt (min): 3.072; Area % -98.042.

[0391] Step 5: 3-(4-((2-(1-(aminomethyl)cyclohexyl)ethyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0392]

[0393] To an ice-cold solution of tert-butyl ((1-(2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)ethyl)cyclohexyl)methyl)carbamate 5 (620 mg, 1.2437 mmol, 1 equiv) in DCM (10 mL) was added HCl in EtOAc (1 M, 6 mL) and stirred at room temperature for 12 h. After completion of the reaction was confirmed by LCMS, the reaction mixture was concentrated under vacuum to afford the crude product, which was co-distilled with Milli-Q HO (2×10 mL) and dried under vacuum to afford 3-(4-((2-(1-(aminomethyl)cyclohexyl)ethyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (480 mg, 98%) as a light yellow solid.

[0394] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),8.05(s,3H),7.43-7.31(m,1H),7.10(d,J=7.20H z,1H),7.01(d,J=7.60Hz,1H),5.12(q,J=5.20Hz,1H),4.39(d,J=17.20Hz,1H),4.24(d,J= 17.60Hz,1H),3.18-2.97(m,2H),2.94-2.89(m,1H),2.79(d,J=5.60Hz,2H),2.63(d,J=17. 20Hz,1H),2.31(q,J=4.40Hz,1H),2.06-2.04(m,1H),1.77(t,J=7.20Hz,2H),1.44(s,10H), 13 C-NMR (100 MHz, DMSO-d6): δ 173.32, 171.43, 168.37, 138.77, 133.32, 131.39, 129.94, 119.19, 116.76, 52.15, 46.70, 45.18, 41.16, 40.56, 34.88, 33.03, 31.64, 31.49, 25.84, 23.21, 21.11. LCMS: 399.1 (M-HCl). Method: Column: Atlantis dC18 (50×4.6) mm 5 μm,

[0395] Mobile phase: A: 0.1% formic acid in H2O; B: ACN; Flow rate: 1.5 mL / min. Rt (min): 1.551; Area % -99.47. HPLC: 99.499%, Rt (min): 8.979.

[0396] Compound 46 in the form of hydrochloride

[0397]

[0398] General procedure for the preparation of compound 6b

[0399]

[0400] To a solution of compound 4 (1.0 g, 2.32 mmol, 1 equivalent) and compound 5b (336 mg, 4.65 mmol, 2.0 equivalents) in DCE / DMF (10 mL) was added acetic acid (1.0 mL), molecular sieves (1.0 g), and stirred at room temperature for 1 hour under a nitrogen atmosphere. NaBH3CN (585 mg, 9.3 mmol, 4 equivalents) was added in batches, and stirred at room temperature (rt or RT) for 16 hours under a nitrogen atmosphere. After the reaction was complete (confirmed by TLC analysis (100% EA) and LCMS), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3 × 20 mL) and brine (20 mL), dried over Na2SO4. The resulting mixture was concentrated and purified by silica gel column chromatography (DCM / EA: 2 / 1) and P-HPLC (acetonitrile / H2O: 40%-60%) to obtain compound 6b (400 mg, 35%) as a white solid.

[0401] TLC: EA=100%

[0402] R f (Compound 4) = 0.5

[0403] R f (Compound 6b) = 0.6

[0404] LC-MS: 487.3[M+1] +

[0405] General procedure for the preparation of compound 46

[0406]

[0407] To a solution of compound 6b (400 mg, 0.873 mmol, 1 eq) in methanol (1 mL) was added HCl / EA (2 N, 8.73 mmol, 10 eq). The resulting solution was stirred at room temperature for 24 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by P-HPLC (acetonitrile / HO: 30%-40%) to afford compound 46 (60 mg, 16%) as a white solid.

[0408] TLC: DCM / methanol = 10 / 1

[0409] R f(Compound 6b) = 0.6

[0410] R f (Compound 46) = 0.2

[0411] LC-MS: 387.25[M+1] +

[0412] 1 H NMR(400MHz,D2O)δ7.84(s,1H),7.70(s,2H),5.07(d,J=8.7Hz,1H),4.55(s,3H),3.51(s,4H),3.18(s,1H ),2.73(d,J=7.9Hz,4H),2.41(d,J=7.5Hz,1H),2.16(s,1H),1.31(dd,J=93.7,45.1Hz,8H),0.65(s,3H).

[0413] Compound 47 in the form of hydrochloride

[0414] 3-(4-((4-Aminobutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0415]

[0416] Step 3: tert-Butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)carbamate

[0417] To a solution of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 4 (3 g, 6.97 mmol, 1 eq) and valeraldehyde (6.0 g, 69.7 mmol, 10 eq) in a mixture of 1,2-dichloroethane (60 mL, 20 vol) and DMF (30 mL, 10 vol) was added acetic acid (1.2 mL, 20.91 mmol, 3 eq) and the reaction mixture was stirred at room temperature for 2 h. NaCNBH (1.75 g, 27.9 mmol, 4 eq) was added portionwise and the reaction mixture was stirred at room temperature for an additional 16 h. TLC analysis (70% EtOAc in petroleum ether) indicated product formation and incomplete consumption of the starting material. Additional amounts of valeraldehyde (2 x 6.0 g, 69.7 mmol, 10 equiv), acetic acid (2 x 0.8 mL, 13.94 mmol, 2 equiv) and NaCNBH3 (0.87 g, 13.94 mmol, 2 equiv) were added and the resulting reaction mixture was stirred at room temperature for 36 hours. The reaction was complete (by TLC analysis (70% EtOAc in petroleum ether, Rf After 4 hours of heating (10 minutes, 1 hour, 1 hour, 3 hours of heating), the reaction mixture was diluted with DCM (100 mL), washed with water (3 x 100 mL), brine (300 mL), dried over anhydrous Na2SO4, filtered and concentrated. The resulting residue was purified by chromatography (Biotage R snapcartridge, KP-Sil, 100 g, 230-400 silica gel) using 65-70% ethyl acetate in petroleum ether to afford tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)carbamate 5 (2.5 g, 97%) as a white solid.

[0418] 1 H-NMR (400MHz, DMSO-d6): δ (s, 1H), 7.37 (t, J = 8.00Hz, 1H), 7.18 (d, J = 7.20H z,1H),7.07(d,J=8.00Hz,1H),6.78(t,J=5.60Hz,1H),5.11(q,J=5.20Hz,1H) ,4.42-4.27(m,2H),3.17(t,J=8.40Hz,4H),2.96-2.90(m,3H),2.68-2.61(m ,2H),2.03-2.00(m,1H),1.43(s,15H),1.38(t,J=10.80Hz,4H),0.86(s,3H).

[0419] LC-MS: 501.3 (M+H), Rt (min): 1.853, Area% -97.842.

[0420] Step 4: 3-(4-((4-aminobutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0421]

[0422] To an ice-cold solution of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)carbamate (2.5 g, 4.99 mmol) in DCM (25 mL) was added HCl (4 M in EtOAc, 12.5 mL) and stirred at room temperature for 2 h. The reaction was complete (by TLC analysis (100% EtOAc R fAfter 5% HCl (confirmed at 0.4 mmol / l), the reaction mixture was concentrated and lyophilized to afford 3-(4-((4-aminobutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride 47 (2.4 g, 98%) as an off-white solid.

[0423] 1 H-NMR (400MHz, DMSO-d6): δ11.01(s,1H),7.92(s,4H),7.39(s,2H),5.15-5.10(m,1H),3.28(s,4H),2.98-2.92(m,1H), 2.75(d,J=5.20Hz,2H),2.68-2.64(m,2H),2.08-2.04(m,1H),1.54(s,6H),1.26(d,J=13.60Hz,5H),0.87-0.81(m,3H).

[0424] LCMS: 401.2 (M+H). Method: Mobile phase A: 0.1% TFA in H2O. Mobile phase B: 0.1% TFA in ACN. Flow rate: 1.5 mL / min. Column: XBridge C8 (50 × 4.6 mm), 3.5 μm. Rt (min): 1.356; Area %: 96.152.

[0425] HPLC: Method: Mobile phase A: 0.1% TFA in H2O. Mobile phase B: acetonitrile. Flow rate: 2.0 mL / min. Column: X-Bridge C8 (50 × 4.6 mm, 3.5 μm). Rt (min): 2.341; Area %: 99.332.

[0426] Compound 48 in the form of hydrochloride

[0427]

[0428] General procedure for the preparation of compound 6d

[0429]

[0430] To a solution of compound 4 (2.0 g, 4.65 mmol, 1 eq.) and compound 5d (932 mg, 9.30 mmol, 2.0 eq.) in DCE / DMF (20 mL) was added acetic acid (2.0 ml) and molecular sieves (1.0 g), and stirred at room temperature under a nitrogen atmosphere for 1 h. NaBH3CN (1.17 g, 18.6 mmol, 4 eq.) was added portionwise, and stirred at room temperature under a nitrogen atmosphere for 16 h. After completion of the reaction (confirmed by TLC analysis (100% EA) and LCMS), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3 x 30 mL) and brine (30 mL), and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (DCM / EA: 2 / 1) and P-HPLC (acetonitrile / H2O: 40%-60%) to obtain compound 6d (660 mg, 28%) as a white solid.

[0431] TLC: EA=100%

[0432] R f (Compound 4) = 0.5

[0433] R f (Compound 6d) = 0.6

[0434] LC-MS: 515.40[M+1] +

[0435] General procedure for the preparation of compound 48

[0436]

[0437] To a solution of compound 6d (660 mg, 1.28 mmol, 1 eq) in DCM (6 mL) was added HCl / EA (2 N, 6.42 mmol, 5 eq). The resulting solution was stirred at 25 ° C for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by P-HPLC (acetonitrile / HO: 30%-40%) to obtain compound 48 (150.55 mg, 28%) as a white solid.

[0438] TLC: DCM / methanol = 10 / 1

[0439] R f (Compound 6d) = 0.6

[0440] R f (Compound 48) = 0.2

[0441] LC-MS: 415.40[M+1] +

[0442] 1 H NMR (400MHz, CDCl3): δ10.25(s,1H),7.75(s,3H),7.40–7.28(m,2H),6.98(s,1H),5.17(d,J=8.9Hz,1H),4.40(dd, J=58.4,16.5Hz,2H),3.13–2.54(m,9H),2.03(d,J=46.6Hz,7H),1.63(s,2H),1.31(d,J=63.7Hz,13H),0.85(s,3H).

[0443] Compound 49 in the form of hydrochloride

[0444]

[0445] General procedure for the preparation of compound 2e

[0446]

[0447] Under a nitrogen atmosphere, at -78 ° C, over a period of 15 minutes, a solution of DMSO (10 mL) in DCM (100 mL) was added dropwise to a solution of oxalyl chloride (9.5 g, 74.5 mmol, 1.2 equivalents) in DCM (50 mL), and stirred at the same temperature for 15 minutes. A solution of compound 1e (10.0 g, 62.1 mmol, 1.0 equivalents) in DCM (50 mL) was added dropwise over a period of 15 minutes. After the reaction mixture was stirred at -78 ° C for 30 minutes, TEA (18.8 g, 188.2 mmol, 3.0 equivalents) was added dropwise at -78 ° C. Once the addition was complete, the reaction mixture was slowly warmed to 0 ° C and stirred for 30 minutes. The reaction was complete (analyzed by TLC (PE / EA=4 / 1, R f After 4% HCl (0.5% NaHCO₄ or ninhydrin) was added, the reaction mixture was diluted with DCM (50 mL). The mixture was washed with 10% citric acid (500 mL), water (50 mL) and brine (50 mL), dried over Na₂SO₄, filtered and concentrated under vacuum below 30° C. to afford compound 2e (11.4 g, crude product) as a yellow oil, which was used without further purification.

[0448] TLC: PE / EA=4:1, KMnO4, ninhydrin

[0449] R f (Compound 1e) = 0.2

[0450] R f (Compound 2e) = 0.4

[0451] LC-MS: none found.

[0452] General procedure for the preparation of compound 5

[0453]

[0454] To a solution of compound 3 (0.2 g, 3.9 mmol, 1.0 equiv) and compound 4 (1.0 g, 3.9 mmol, 1.0 equiv) in DCE / DMF (1 / 1, 20 mL) was added acetic acid (0.7 g, 11.6 mmol, 3.0 equiv). The reaction mixture was stirred at room temperature for 2 h. At this stage, NaBH3CN (1.0 g, 15.4 mmol, 4.0 equiv) was added portionwise at 0 ° C and stirred at room temperature for 18 h. After the reaction was completed, the reaction mixture was diluted with DCM (100 mL). The mixture was washed with water (2 × 20 mL) and brine (2 × 20 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (DCM / methanol: 3: 1) to obtain compound 5 (0.75 g, 64%) as a white solid.

[0455] TLC: EA

[0456] R f (Compound 3) = 0.4

[0457] R f (Compound 5) = 0.5

[0458] LC-MS: 302.25[M+1] + ,300.25[M-1] -

[0459] 1 H NMR (400MHz, CDCl3): δ7.10(d,J=5.9Hz,1H),6.70(d,J=6.8Hz,1H),5.06(d,J=10.9Hz,1H),4.19(d,J=15.7Hz,1H),4.07(d,J =15.7Hz,1H),3.29(d,J=24.2Hz,1H),3.07(s,2H),2.75(s,2H),2.25(s,1H),2.09(s,1H),1.57(d,J=6.1Hz,2H),0.91(s,3H).

[0460] 1H NMR (400 MHz, DMSO-d6): δ 9.33 (d, J = 1.8 Hz, 1H), 8.47–8.39 (m, 2H), 8.21 (d, J = 7.4 Hz, 1H), and 8.14 (d, J = 8.4 Hz, 1H).

[0461] General procedure for the preparation of compound 6e

[0462]

[0463] To a solution of compound 5 (4.6 g, 15.3 mmol, 1.0 equivalent) and compound 2e (4.4 g, 27.5 mmol, 1.8 equivalent) in DCE / DMF (80 mL, 1 / 1) was added acetic acid (20 mL) and stirred at room temperature for 1 h under a nitrogen atmosphere. NaBH CN (3.8 g, 61.1 mmol, 4.0 equivalent) was added in portions and stirred at room temperature for 18 hours under a nitrogen atmosphere. After completion of the reaction (confirmed by TLC analysis (100% EA) and LCMS), the reaction mixture was diluted with DCM (400 mL). The mixture was washed with water (2 × 40 mL) and brine (2 × 40 mL) and dried over Na SO. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (DCM / EA: 2: 1) and P-HPLC (acetonitrile / H O: 40%-60%) to obtain compound 6e (0.7 g, 10%) as a white solid.

[0464] TLC: EA=100%

[0465] R f (Compound 5) = 0.5

[0466] R f (Compound 6e) = 0.6

[0467] LC-MS:445[M+1] + ,443[M-1] -

[0468] 1 H NMR (400MHz, CDCl3): δ8.14(s,1H),7.42(d,J=31.6Hz,2H),7.12(s,1H),5.22(d,J=10.3Hz,1H),4.45(s,2H),3.48 (s,1H),3.17(d,J=51.0Hz,5H),2.97–2.75(m,2H),2.21(s,1H),1.67(s,2H),1.42(d,J=17.8Hz,14H),0.87(s,3H).

[0469] General procedure for the preparation of compound 49

[0470]

[0471] To a solution of compound 6e (0.2 g, 0.4 mmol, 1.0 equiv) in methanol (2 mL) was added HCl / EA (2 mL, 2 N, 4.0 mmol, 10.0 equiv). The resulting solution was stirred at room temperature for 18 h. The resulting mixture was concentrated under reduced pressure and purified by P-HPLC (acetonitrile / HO: 30%-40%) to afford compound 49 (147 mg, 95%) as a white solid.

[0472] LC-MS: 345.35[M+1] +

[0473] 1 H NMR (400MHz, D2O): δ7.42–7.30(m,2H),7.24(d,J=7.5Hz,1H),5.00(dd,J=12.9,4.4Hz,1H),4.43(q,J=17.3Hz,2H),3.41(d,J=5.4Hz,2H),3.06– 2.93(m,4H),2.75(dd,J=25.3,11.4Hz,2H),2.41(dd,J=12.7,4.6Hz,1H),2.17–2.07(m,1H),1.28(dd,J=14.2,7.0Hz,2H),0.65(t,J=7.1Hz,3H).

[0474] Compound 50

[0475] N-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(propyl)amino)butyl)acetamide

[0476]

[0477] Step 1: N-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(propyl)amino)butyl)acetamide

[0478]

[0479] To a solution of 3-(4-((4-aminobutyl)(propyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl compound 4 (200 mg, 0.489 mmol, 1 eq) in pyridine (3 mL) at 0 °C was added acetic anhydride (0.231 mL, 2.445 mmol, 5 eq) and allowed to stir at room temperature for 12 h. After completion of the reaction (confirmed by LCMS), the reaction mixture was concentrated under vacuum to provide the crude compound, which was purified by reverse phase column chromatography (Grace column: C18 40 μm, 40 g; flow rate: 20 mL / min; 0.1% aqueous HCOOH / ACN mobile phase) and the pure fractions were lyophilized to afford N-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(propyl)amino)butyl)acetamide (86 mg, 0.207 mmol, 43.0% yield) as a yellow solid.

[0480] 1 H-NMR (400MHz, DMSO-d6): δ10.97(s,1H),7.77(t,J=5.20Hz,1H),7.38(t,J=7.6 0Hz,1H),7.18(t,J=0.40Hz,1H),7.07(d,J=7.60Hz,1H),5.10(q,J=5.20Hz,1H), 4.35(q,J=16.80Hz,2H),3.21-3.13(m,4H),3.00(q,J=6.40Hz,4H),2.57(s,1H) ,2.02(q,J=5.20Hz,1H),1.76(s,3H),1.48-1.36(m,6H),0.84(t,J=7.60Hz,3H).

[0481] LCMS: 415.1 (M+H), method: mobile phase: A: 0.1% FA in H2O, mobile phase: B: ACN, column: Atlantis dC18 (50×4.6 mm) 5 μm, flow rate: 1.5 mL / min Rt (min): 1.622, area %: 98.693.

[0482] HPLC: Method: Mobile phase: A: 0.1% FA in water, Mobile phase: B: ACN, Column: X-Bridge C8 (50×4.6) mm, 3.5 μm, Flow rate: 2.0 mL / min, Rt (min): 1.789, Area %: 99.953.

[0483] The hydrochloride form of compound 51

[0484]

[0485] General procedure for the preparation of compound 2g

[0486]

[0487] To a solution of compound 1g (5.0g, 48mmol, 1 equivalent) and tert-butylchlorodiphenylsilane (13.2g, 48mmol, 1.0 equivalent) in DMF (50mL) was added DIEA (18.6g, 144mmol, 3.0 equivalents) and stirred at room temperature for 16h under a nitrogen atmosphere. After completion of the reaction (confirmed by TLC analysis (PE / EA=2:1) ​​and LCMS), the reaction mixture was diluted with DCM (20mL). The mixture was washed with water (3×20mL) and brine (20mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA:10 / 1) to obtain compound 2g (7.2g, 44%) as a clear oil.

[0488] TLC: PE / EA=2 / 1KMnO4

[0489] R f (Compound 1g) = 0.1

[0490] R f (Compound 2g) = 0.6

[0491] 1 H NMR (400MHz, CDCl3): δ7.66(d,J=5.7Hz,4H),7.38(d,J=6.8Hz,6H),3.64(dd ,J=17.5,4.3Hz,4H),1.63–1.50(m,5H),1.43(d,J=6.3Hz,2H),1.04(s,10H).

[0492] General procedure for the preparation of compound 3g

[0493]

[0494] Under nitrogen atmosphere, at -78 ℃, over 15 minutes, to a solution of oxalyl chloride (3.2 g, 25.2 mmol, 1.2 equivalents) in DCM (40 mL) was added dropwise a solution of DMSO (7.5 mL, 1.0 V) in DCM (35 mL), and stirred at the same temperature for 15 minutes. At -78 ℃, a solution of compound 2g (7.2 g, 20.9 mmol, 1.0 equivalents) in DCM (20 mL) was added dropwise over 15 minutes. After the reaction mixture was stirred at -78 ℃ for 30 minutes, TEA (6.35 g, 62.9 mmol, 3.0 equivalents) was added dropwise at -78 ℃. Once the addition was complete, the reaction mixture was slowly warmed to 0 ℃ and stirred for 30 minutes. After completion of the reaction (confirmed by TLC analysis), the reaction mixture was diluted with DCM (50 mL) and washed with 10% citric acid (30 mL), water (50 mL), brine (50 mL), and dried over Na SO The solution was filtered and concentrated under vacuum below 30° C. to afford Compound 3g (7.03 g) as a clear oil which was used without further purification.

[0495] TLC: PE / EA=2 / 1

[0496] R f (Compound 2g) = 0.6

[0497] R f (Compound 3g) = 0.8

[0498] 1 H NMR (400MHz, CDCl3): δ9.74 (s, 1H), 7.66 (d, J = 7.3Hz, 4H), 7.48–7.33 (m, 6H), 3.67 (t, J = 6.1 Hz,2H),2.41(t,J=7.3Hz,2H),1.79–1.68(m,2H),1.60(dd,J=14.3,6.6Hz,3H),1.05(s,9H).

[0499] General procedure for the preparation of compound 5g

[0500]

[0501] To a solution of compound 4a (2.2 g, 5.1 mmol, 1 equivalent) and compound 3g (7.03 g, 20.6 mmol, 4.0 equivalents) in DCE / DMF (10 mL) was added acetic acid (2.2 mL) and molecular sieves (7.03 g) and stirred at room temperature under a nitrogen atmosphere for 1 h. NaBH3CN (1.6 g, 25.6 mmol, 5 equivalents) was added in portions and stirred at room temperature under a nitrogen atmosphere for 16 h. After completion of the reaction (confirmed by TLC analysis (100% EA) and LCMS), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3 x 20 mL) and brine (20 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA: 2 / 1) and P-HPLC (acetonitrile / H2O: 40%-60%) to obtain compound 5g (1.03 g, 27%) as a white solid.

[0502] TLC: EA=100%

[0503] R f (Compound 4a) = 0.5

[0504] R f (Compound 5g) = 0.7

[0505] LC-MS:755.60[M+1] +

[0506] General procedure for the preparation of compound 6g

[0507]

[0508] To compound 5g (1.03g, 1.37mmol, 1 equivalent) solution in THF, add TBAF / THF (13.7mL, 1N, 13.7mmol, 10 equivalents), and under nitrogen atmosphere, at room temperature, stir 4h. After reaction is complete (by TLC analysis and LCMS confirmation), reaction mixture is diluted with DCM (20mL). The mixture is washed with water (3 × 20mL) and brine (20mL), through Na SO Drying. The gained mixture is concentrated under reduced pressure and passed through silica gel column chromatography (PE / EA: 2 / 1) and P-HPLC (acetonitrile / H O: 40%-60%) purifying, to obtain compound 6g (580mg, 82%) as white solid.

[0509] TLC: EA=100%

[0510] R f (Compound 5g) = 0.7

[0511] R f(Compound 6g) = 0.4

[0512] LC-MS:517.30[M+1] +

[0513] General procedure for the preparation of compound 51

[0514]

[0515] To a solution of compound 6g (200 mg, 0.387 mmol, 1 eq) in methanol (1.0 mL) was added HCl (2.0 mL, 2N, 4.0 mmol, 10.0 eq). The resulting solution was stirred at room temperature for 24 h. The reaction was lyophilized to afford compound 51 (164.3 mg, 87%) as a white solid.

[0516] TLC: DCM / methanol = 10 / 1

[0517] R f (Compound 6g) = 0.4

[0518] R f (Compound 51) = 0.2

[0519] LC-MS:417.40[M+1] +

[0520] 1 H NMR (400MHz, D2O): δ7.88(d,J=7.2Hz,1H),7.77(d,J=7.7Hz,1H),7.70(t,J=7.6Hz,1H),5.10–5.01(m,1H),4.53(t,J=13.1Hz,3H ),3.56(d,J=6.9Hz,4H),3.32(t,J=5.7Hz,2H),2.76(dd,J=27.9,6.5Hz,4H),2.47–2.32(m,1H),2.16(s,1H),1.54–1.07(m,11H).

[0521] Compound 52 in the form of hydrochloride

[0522]

[0523] General procedure for preparing compound 2h

[0524]

[0525] Under nitrogen atmosphere, a solution of compound 1h (0.5 g, 3.47 mmol, 1.0 eq) in THF (5 mL) was added dropwise at 0° C. to a solution of LiAlH 4 (145 mg, 3.82 mmol, 1.1 eq) in THF (5 mL), and then stirred at room temperature for 4 h. The reaction was complete (analyzed by TLC (PE / EA=2 / 1, R f After 4% HCl (confirmed with KMnO), the reaction mixture was diluted with MTBE (30 mL) and NaSO·10H O was added. The mixture was stirred for 10 minutes, filtered and concentrated to afford compound 2h (350 mg, 77%) as a clear oil, which was used without further purification.

[0526] TLC: PE / EA=2:1, KMnO4

[0527] R f (Compound 1h) = 0.2

[0528] R f (Compound 2h) = 0.6

[0529] 1 H NMR (400MHz, CDCl3): δ3.62(t,J=5.7Hz,2H),1.59–1.44(m,3H),1.28(d,J=15.7Hz,5H),1.17(s,3H),0.85(d,J=5.6Hz,7H).

[0530] General procedure for the preparation of compound 3h

[0531]

[0532] To a solution of compound 2h (600 mg, 4.61 mmol, 1 eq) in DCM (30 mL) was added PCC (1.99 g, 9.22 mmol, 2.0 eq) and stirred at room temperature for 4 h. After completion of the reaction (confirmed by TLC analysis (PE / EA=3:1)), silica gel (4.04 g) was added to the reaction mixture and stirred for an additional 10 minutes. The reaction mixture was filtered and concentrated under reduced pressure below 30 ° C to obtain compound 3h (714 mg) as a clear oil, which was used without further purification.

[0533] TLC: PE / EA=4 / 1KMnO4

[0534] R f (Compound 2h) = 0.4

[0535] R f (Compound 3h) = 0.7

[0536] 1 H NMR (400MHz, CDCl3): δ9.75 (s, 1H), 2.41 (s, 2H), 1.56 (d, J = 29.1Hz, 3H), 1.24 (d, J = 50.5Hz, 5H), 0.85 (s, 6H).

[0537] General procedure for the preparation of compound 5h

[0538]

[0539] To a solution of compound 4a (799 mg, 1.86 mmol, 1 eq) and compound 3h (714 mg, 5.57 mmol, 3.0 eq) in DCE / DMF (6 mL) was added acetic acid (1 mL) and molecular sieves (1.0 g), and stirred at room temperature for 1 h under a nitrogen atmosphere. NaBH3CN (467 mg, 7.43 mmol, 4 eq) was added portionwise at room temperature under a nitrogen atmosphere and stirred for 16 h. After completion of the reaction (confirmed by TLC analysis (100% EA) and LCMS), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3×20 mL) and brine (20 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (DCM / EA: 2 / 1) and P-HPLC (acetonitrile / HO: 40%-60%) to obtain compound 5h (275 mg, 27%) as a white solid.

[0540] TLC: EA=100%

[0541] R f (Compound 4a) = 0.4

[0542] R f (Compound 5h) = 0.6

[0543] LC-MS:543.55[M+1] +

[0544] General procedure for the preparation of compound 52

[0545]

[0546] To a solution of compound 5h (200 mg, 0.368 mmol, 1.0 eq) in methanol (1.0 mL) was added HCl (1.8 mL, 2 N, 3.7 mmol, 10.0 eq). The resulting solution was stirred at room temperature for 24 h. The reaction mixture was lyophilized to afford compound 52 (170.7 mg, 89%) as a white solid.

[0547] TLC: DCM / methanol = 10 / 1

[0548] R f (Compound 5h) = 0.6

[0549] R f (Compound 52) = 0.1

[0550] LC-MS:443.45[M+1] +

[0551] 1 H NMR (400MHz, D2O): δ7.84(d,J=6.9Hz,1H),7.71(dd,J=21.3,7.2Hz,2H),5.04(d,J=9.6Hz,1H),4.51(t,J=13.0Hz,2H),3.51(s,4H),2.7 3(t,J=14.1Hz,4H),2.39(d,J=9.7Hz,1H),2.14(s,1H),1.31(dd,J=76.5,36.4Hz,7H),1.02(s,4H),0.82(s,2H),0.57(d,J=5.5Hz,6H).

[0552] Compound 53

[0553]

[0554] General procedure for the preparation of compound 2i

[0555]

[0556] Under a nitrogen atmosphere, at -78 ° C, over a period of 15 minutes, a solution of DMSO (1 mL) in DCM (10 mL) was added dropwise to a solution of oxalyl chloride (0.9 g, 6.9 mmol, 1.2 equivalents) at DCM (5 mL), and stirred at the same temperature for 15 minutes. A solution of compound 1i (1.0 g, 5.7 mmol, 1.0 equivalents) in DCM (5 mL) was added dropwise over a period of 15 minutes. After the reaction mixture was stirred at -78 ° C for 30 minutes, TEA (1.7 g, 17.1 mmol, 3.0 equivalents) was added dropwise at -78 ° C. Once the addition was complete, the reaction mixture was slowly warmed to 0 ° C and stirred for 30 minutes. The reaction was complete (analyzed by TLC (PE / EA=4 / 1, R fAfter 4% HCl (0.5% NaHCO₄ or ninhydrin) was added, the reaction mixture was diluted with DCM (10 mL). The mixture was washed with 10% citric acid (10 mL), water (10 mL) and brine (10 mL), dried over Na₂SO₄, filtered and concentrated under vacuum below 30° C. to afford compound 2i (0.9 g, crude product) as a yellow oil, which was used without further purification.

[0557] TLC: PE / EA=4:1, KMnO4, ninhydrin

[0558] R f (Compound 1i) = 0.2

[0559] R f (Compound 2i) = 0.4

[0560] LC-MS: none found.

[0561] 1 H NMR (400MHz, CDCl3): δ9.79 (s, 1H), 4.89 (s, 1H), 3.39 (s, 2H), 2.64 (d, J = 35.8Hz, 2H), 1.40 (s, 10H).

[0562] General procedure for the preparation of compound 6i

[0563]

[0564] To a solution of compound 2i (2.3 g, 13.3 mmol, 2.0 equivalents) and compound 5 (2.0 g, 6.6 mmol, 1.0 equivalents) in DCE / DMF (1 / 1, 40 mL) was added acetic acid (10 mL) and stirred at room temperature under a nitrogen atmosphere for 1 h. NaBH CN (1.7 g, 26.6 mmol, 4.0 equivalents) was added in portions and stirred at room temperature under a nitrogen atmosphere for 18 h. After completion of the reaction (confirmed by TLC analysis (100% EA) and LCMS), the reaction mixture was diluted with DCM (200 mL). The mixture was washed with water (2 × 20 mL) and brine (2 × 20 mL) and dried over Na SO. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (DCM / methanol: 2 / 1) and P-HPLC (acetonitrile / H O: 40%-60%) to obtain compound 6i (0.4 g, 26%) as a white solid.

[0565] TLC: EA

[0566] R f (Compound 2i) = 0.5

[0567] Rf (Compound 6i) = 0.6

[0568] LC-MS:459[M+1] + ,457.30[M-1] -

[0569] General procedure for the preparation of compound 53

[0570]

[0571] To a solution of compound 6i (0.2 g, 0.4 mmol, 1.0 equiv) in DCM / H2O (1 / 1, 2 mL) was added TFA (4 mmol, 10.0 equiv). The resulting solution was stirred at room temperature for 24 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by P-HPLC eluting with acetonitrile / H2O (30%-40%) to afford compound 53 (105 mg, 65%) as a white solid.

[0572] LC-MS:359.30[M+1] + ,357.20[M-1] -

[0573] 1 H NMR (400MHz, CD3OD): δ7.51–7.36(m,2H),7.29(d,J=7.5Hz,1H),5.14(dd,J= 12.9,4.2Hz,1H),4.46(s,2H),3.31(d,J=9.3Hz,5H),3.14(d,J=6.5Hz,2H),2 .92(dd,J=19.2,11.4Hz,3H),2.78(d,J=17.2Hz,1H),2.54(d,J=9.8Hz,1H), 2.19(d,J=5.7Hz,1H),1.82(s,2H),1.57–1.44(m,2H),0.88(t,J=6.8Hz,3H).

[0574] Compound 55 in the form of hydrochloride

[0575] 3-(4-((4-Aminobutyl)(5-aminopentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[0576]

[0577] Step 1: tert-Butyl (5-((tert-Butyldimethylsilyl)oxy)pentyl)carbamate

[0578]

[0579] To a solution of tert-butyl (5-hydroxypentyl)carbamate 1 (1.0 g, 4.92 mmol, 1.0 eq) in DCM (20 mL) was added imidazole (0.837 g, 12.30 mmol, 2.5 eq) and TBS-Cl (1.112 g, 7.38 mmol, 1.5 eq) at 0°C and allowed to stir at room temperature for 2 h. The reaction was complete (by TLC (50% EtOAc / petroleum ether, R f After 4 hours, 1% t-butyl-5-((tert-butyldimethylsilyl)oxy)pentyl)carbamate 2 (1.5 g, 4.70 mmol, 96% yield) was obtained. The reaction mixture was quenched with H2O (20 mL) and extracted with DCM (2 x 20 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum to afford the crude product (tert-butyl 5-((tert-butyldimethylsilyl)oxy)pentyl)carbamate 2 (1.5 g, 4.70 mmol, 96% yield) as a yellow liquid, which was used further without purification. 1 H-NMR (400MHz, DMSO-d6): δ6.74(s,1H),3.54-3.56(m,2H),2.88-2.90(m,2H),1.37-1.43(m,6H),1.156(s,9H),0.86(s,9H),0.03(s,6H).

[0580] Step 2: tert-Butyl (tert-butoxycarbonyl)(5-((tert-butyldimethylsilyl)oxy)pentyl)carbamate

[0581]

[0582] At 0 ℃, to a stirred solution of (5-((tert-butyldimethylsilyl)oxy)pentyl)t-butyl carbamate 2 (1.5g, 4.72mmol, 1.0 equivalent) in THF (20mL), n-BuLi (2.5M hexane solution, 2.267mL, 5.67mmol, 1.2 equivalents) was added, and the reaction mixture was stirred for 15min at 0 ℃. (Boc) was added to the reaction mixture at the same temperature. The resulting reaction mixture was warmed to room temperature and stirred for 1 hour. The reaction was completed (by TLC analysis (10% EtOAc / petroleum ether, R fAfter 4 hours, 1% t-butyl-2-oxo-1-oxo-4-oxo-6-nitropropane (1% t-butyl-2-oxo-1-oxo-4-nitropropane) was added to the mixture of 4, 2, 3, 4, 5-tetramethyl-2-oxo-1-oxo-4-nitropropane (1% t-butyl-2-oxo-1-oxo-4-nitropropane) and 4, 2, 3, 4, 5-tetramethyl-2-oxo-1-oxo-4-nitropropane) (1% t-butyl ...

[0583] 1 H-NMR (400MHz, DMSO-d6): δ3.55-3.58(m,2H),3.44-3.48(m,2H),1.25-1.43(m,6H),1.12(s,18H),0.88(s,9H),0.03(s,6H)

[0584] Step 3: tert-Butyl (tert-butoxycarbonyl)(5-hydroxypentyl)carbamate

[0585]

[0586] To a stirred solution of tert-butyl (tert-butoxycarbonyl)(5-((tert-butyldimethylsilyl)oxy)pentyl)carbamate 3 (1.9 g, 4.55 mmol) in THF (20 mL) was added TBAF (1 M, 9.10 mL, 9.10 mmol, 2.0 equiv) at room temperature. The resulting solution was stirred at room temperature overnight. The reaction was complete (by TLC analysis (20% EtOAc / petroleum ether, R f To the 4-thiazolinyl group (1.0g, 3.27mmol, 71.9% yield) of tert-butyl (tert-butoxycarbonyl) (5-hydroxypentyl) carbamate 4 (1.0g, 3.27mmol, 71.9% yield) of the petroleum ether solution of 15-20% EtOAc was added 4-thiazolinyl group (1.0g, 3.27mmol, 71.9% yield) of the ...

[0587] 1H-NMR (400MHz, DMSO-d6): δ4.36 (t, J=4.80Hz, 1H), 3.40-3.47 (m, 2H), 3.37-3.4 0(m,2H),1.48-1.50(m,2H),1.44(s,18H),1.40-1.42(m,2H),1.25-1.27(m,2H).

[0588] Step 4: tert-Butyl (tert-butoxycarbonyl)(5-oxopentyl)carbamate

[0589]

[0590] At-78 ℃, to oxalyl chloride (0.577mL, 6.59mmol, 2.0 equivalents) and CH cl (20mL) solution in add DMSO (0.936mL, 13.18mmol, 4.0 equivalents) at CH cl (5mL) solution in, and stirred 15 minutes at the same temperature.At-78 ℃, add (tert-butoxycarbonyl) (5-hydroxypentyl) t-butyl carbamate 4 (1.0g, 3.30mmol, 1.0 equivalents) at CH cl (5mL) solution in.-78 ℃ of stirring reaction mixture after 30 minutes, at-78 ℃, dropwise add triethylamine (2.76mL, 19.78mmol, 6.0 equivalents).Then, reaction mixture is slowly warmed to 0 ℃ and stirs 30min.Reaction is completed (analyze (20% EtOAc / petroleum ether, R by TLC)). f ~0.6, KMnO4) confirmed), the reaction mixture was diluted with DCM (40 mL) and washed with 10% aqueous citric acid (1×40 mL) and water (1×40 mL), dried over Na2SO4, filtered and concentrated under vacuum (below 30°C) to give tert-butyl (tert-butoxycarbonyl)(5-oxopentyl)carbamate 5 (0.95 g, 3.08 mmol, 93% yield) as a light yellow liquid, which was used further without purification.

[0591] 1 H-NMR (400MHz, DMSO-d6): δ0.00(s,1H),3.47(m,2H),2.42-2.46(m,2H),1.47-1.50(m,4H),1.42(s,18H).

[0592] Step 5: tert-Butyl (tert-butoxycarbonyl)(5-((4-((tert-butoxycarbonyl)amino)butyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)pentyl)carbamate

[0593]

[0594] To a stirred mixture of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 6 (0.7 g, 1.626 mmol, 1.0 equiv) and tert-butyl (tert-butoxycarbonyl)(5-oxopentyl)carbamate 5 (0.980 g, 3.25 mmol, 2.0 equiv) in CHCl (10 mL) and DMF (10 mL) was added trifluoroacetic acid (0.501 mL, 6.50 mmol, 4.0 equiv) and stirred at 0°C for 10 min. Sodium triacetoxyborohydride (1.378 g, 6.50 mmol, 4.0 equiv) was added portionwise at 0°C and the resulting colorless turbid material was stirred at room temperature for 16 h. The reaction was complete (by TLC analysis (100% EtOAc, R f To the 4-thiazolinyl group (0.70g, 0.891mmol, 54.8% yield) of tert-butyl (tert-butoxycarbonyl)(5-((4-((tert-butoxycarbonyl)amino)butyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)pentyl)carbamate 7 (0.65g, 0.891mmol, 54.8% yield) was added 4-thiazolinyl group (0.70g, 0.891mmol, 54.8% yield) of ...

[0595] 1 H-NMR (400MHz, DMSO-d6): δ0.00(s,1H),7.35-7.39(m,1H),7.18-7.20(m,1H),7 .06-7.08(m,1H),6.75-6.78(m,1H),5.08-5.13(m,1H),4.27-4.41(m,2H),3.45- 3.46(m,2H),2.89-2.92(m,4H),2.74-2.74(m,4H),2.51-2.52(m,2H),2.50-2.50 (m,1H),2.00-2.03(m,1H),1.45-1.49(m,15H),1.42(s,18H),1.20-1.26(m,2H),

[0596] LCMS: 616.4 (M-Boc). Method: Atlantis dC18 (50×4.6 mm) 5 μm, mobile phase: A: 0.1% FA in H2O, mobile phase: B: ACN, flow rate: 1.5 mL / min

[0597] HPLC: Method: Mobile phase A: 0.1% TFA in water, Mobile phase B: acetonitrile, Flow rate: 2.0 mL / min. Column: X-Bridge C8 (50×4.6) mm, 3.5 μm. RT: 4.972 min, Area: 98.095%

[0598] Step 6: 3-(4-((4-aminobutyl)(5-aminopentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[0599]

[0600] To a stirred solution of tert-butyl (tert-butoxycarbonyl)(5-((4-((tert-butoxycarbonyl)amino)butyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)pentyl)carbamate 7 (0.35 g, 0.489 mmol, 1.0 equiv) in dichloromethane (5 mL) was added HCl (4 M in ethyl acetate, 10 mL) at 0°C and then stirred at room temperature for 3 h. The reaction progress was monitored by LCMS. The reaction mixture was concentrated under vacuum at below 40°C to afford a light yellow solid. The obtained solid was dissolved in water (10 mL) and washed with MTBE (2×10 mL), and the aqueous layer was lyophilized to afford 3-(4-((4-aminobutyl)(5-aminopentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl (0.2 g, 0.404 mmol, 83% yield) as a light yellow solid.

[0601] 1 H-NMR (400MHz, CD3OD): δ8.20(s,2H),7.83(s,1H),5.22-5.27(m,1H),4.84-4.88(m,2H),3.72(m,4 H),2.87-2.97(m,6H),2.63-2.66(m,1H),2.27-2.28(m,1H),1.60-1.70(m,7H),1.42-1.47(m,3H).

[0602] LCMS: 416.3 (M+H). Method: Column: XBridge C8 (50×4.6 mm) 3.5 μm, Mobile phase: A: 0.1% TFA in H 2 O, Mobile phase: B: 0.1% TFA in acetonitrile, Flow rate: 1.5 mL / min.

[0603] HPLC: Method: Mobile phase A: 0.1% TFA in water, Mobile phase B: Methanol, Flow rate: 0.7 mL / min. Column: Atlantis dC18 (250×4.6) mm, 5 μm. RT: 12.223 min, Area: 98.472%.

[0604] Compound 56 in the form of hydrochloride

[0605]

[0606] General procedure for the preparation of compound 21

[0607]

[0608] To a solution of compound 11 (10 g, 85.6 mmol, 1 eq) and tert-butylchlorodiphenylsilane (23.25 g, 85.6 mmol, 1.0 eq) in DMF (100 mL) was added DIEA (32.7 g, 254 mmol, 3.0 eq) and stirred at room temperature under a nitrogen atmosphere for 16 h. After completion of the reaction (confirmed by TLC analysis (PE / EA=2:1) ​​and LCMS), the reaction mixture was diluted with DCM (20 mL) and washed with water (3×20 mL) and brine (20 mL), dried over Na 2 SO 4, and filtered. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA: 10 / 1) to obtain compound 21 (10.27 g, 35%) as a clear oil.

[0609] TLC: PE / EA=2 / 1KMnO4

[0610] R f (Compound 11) = 0.1

[0611] R f (Compound 21) = 0.6

[0612] 1 H NMR (400MHz, CDCl3): δ7.67(d,J=5.6Hz,4H),7.39(d,J=6.8Hz,6H),3.72–3.56(m,4H),1.58(t,J=14.0Hz,5H),1.31(d,J=37.5Hz,6H),1.05(s,9H).

[0613] General procedure for the preparation of compound 31

[0614]

[0615] Under nitrogen atmosphere, at -78 ℃, over 15 minutes, to a solution of oxalyl chloride (2.14g, 16.8mmol, 1.2 equivalents) in DCM (20mL), a solution of DMSO (5mL, 1.0V) in DCM (20mL) was added dropwise, and stirred at the same temperature for 15 minutes. Over 15 minutes, a solution of compound 21 (5.0g, 14.02mmol, 1.0 equivalents) in DCM (10mL) was added dropwise. After the reaction mixture was stirred at -78 ℃ for 30 minutes, TEA (4.3g, 42.1mmol, 3.0 equivalents) was added dropwise at -78 ℃. Once the addition was complete, the reaction mixture was slowly warmed to 0 ℃ and stirred for 30 minutes. After the reaction was complete (confirmed by TLC analysis), the reaction mixture was diluted with DCM (50mL). The mixture was washed with 10% citric acid (30 mL), water (50 mL) and brine (50 mL), dried over Na 2 SO 4 , filtered and concentrated under vacuum below 30° C. to afford compound 31 (4.9 g) as a clear oil, which was used without further purification.

[0616] TLC: PE / EA=2 / 1

[0617] R f (Compound 21) = 0.6

[0618] R f (Compound 31) = 0.8

[0619] General procedure for the preparation of compound 51

[0620]

[0621] To a solution of compound 4a (1.5 g, 3.45 mmol, 1 eq) and compound 3l (4.9 g, 13.8 mmol, 4.0 eq) in DCE / DMF (15 mL) was added acetic acid (1.5 mL) and molecular sieves (5.0 g) and stirred at room temperature under a nitrogen atmosphere for 1 h. NaBH3CN (1.08 g, 17.2 mmol, 5 eq) was added portionwise and stirred at room temperature under a nitrogen atmosphere for 16 h. After completion of the reaction (confirmed by TLC analysis (100% EA) and LCMS), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3 x 20 mL) and brine (20 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA: 2 / 1) and P-HPLC (acetonitrile / HO: 40%-60%) to obtain compound 5l (1.03 g, 29%) as a white solid.

[0622] TLC: EA=100%

[0623] R f (Compound 4a) = 0.5

[0624] R f (Compound 51) = 0.7

[0625] LC-MS:769.60[M+1] +

[0626] General procedure for the preparation of compound 61

[0627]

[0628] To a solution of compound 51 (1.03 g, 1.34 mmol, 1 equivalent) in THF was added TBAF / THF (13.4 mL, 1 N, 13.4 mmol, 10 equivalents), and stirred at room temperature for 4 h under a nitrogen atmosphere. After completion of the reaction (confirmed by TLC analysis and LCMS), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3 × 20 mL) and brine (20 mL), dried over Na SO dried. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA: 2 / 1) and P-HPLC (acetonitrile / H O: 40%-60%) to obtain compound 61 (620 mg, 87%) as a white solid.

[0629] TLC: EA=100%

[0630] R f (Compound 51) = 0.7

[0631] R f(Compound 6l)=0.4

[0632] LC-MS:531.45[M+1] +

[0633] General procedure for the preparation of compound 56

[0634]

[0635] To a solution of compound 61 (300 mg, 0.566 mmol, 1 eq) in methanol (1 mL) was added HCl (2.8 mL, 2 N, 5.6 mmol, 10 eq). The resulting solution was stirred at room temperature for 24 h. The reaction mixture was lyophilized to afford compound 56 (231 mg, 81%) as a white solid.

[0636] TLC: DCM / methanol = 10 / 1

[0637] R f (Compound 6l)=0.4

[0638] R f (Compound 56) = 0.2

[0639] LC-MS:431.40[M+1] +

[0640] 1 H NMR (400MHz, D2O): δ7.88(d,J=7.0Hz,1H),7.74(dd,J=22.2,7.6Hz,2H),5.07(d,J=8.9Hz,1H),4.54(t,J=13.1Hz,2H),3.55(d ,J=5.1Hz,4H),3.34(s,2H),2.87–2.67(m,4H),2.41(d,J=8.3Hz,1H),2.17(s,1H),1.35(dd,J=63.6,23.7Hz,9H),1.10(s,4H).

[0641] Compound 57 in the form of hydrochloride

[0642]

[0643] General procedure for the preparation of compound 2m

[0644]

[0645] To a solution of compound 1m (14.3 g, 138.6 mmol, 1.0 eq) and TEA (42.0 g, 415.8 mmol, 3.0 eq) in DCM (150 mL) was added dropwise a solution of (Boc) O (36.3 g, 166.3 mmol, 1.2 eq) followed by addition of DMAP (1.7 g, 13.9 mmol, 0.1 eq) and stirred at room temperature for 18 h. The reaction mixture was diluted with DCM (300 mL) and washed with brine (3 x 50 mL), dried over NaSO, filtered and concentrated under vacuum to afford compound 2m (7.1 g, 24%) as a colorless oil which was used further.

[0646] TLC: PE / EA=4:1, KMnO4, ninhydrin

[0647] R f (Compound 1m) = 0.2

[0648] R f (Compound 2m) = 0.4

[0649] LC-MS: none found.

[0650] General procedure for the preparation of compound 3m

[0651]

[0652] Under a nitrogen atmosphere, at -78 ° C, over a period of 15 minutes, a solution of oxalyl chloride (5.4 g, 42.4 mmol, 1.2 equivalents) in DCM (35 mL) was added dropwise to a solution of DMSO (7 mL, 1.0 V) in DCM (70 mL), and stirred at the same temperature for 15 minutes. A solution of compound 2m (7.1 g, 35.3 mmol, 1.0 equivalents) in DCM (35 mL) was added dropwise over a period of 15 minutes. After the reaction mixture was stirred at -78 ° C for 30 minutes, TEA (10.7 g, 105.9 mmol, 3.0 equivalents) was added dropwise at -78 ° C. Once the addition was complete, the reaction mixture was slowly warmed to 0 ° C and stirred for 30 minutes. The reaction was complete (analyzed by TLC (PE / EA=4 / 1, R f After the reaction mixture was diluted with DCM (70 mL) and the mixture was washed with 10% citric acid solution (35 mL), water (35 mL) and brine (35 mL), and dried over NaSO. The resulting mixture was filtered and concentrated under vacuum at below 30°C to afford compound 3m (7.4 g) as a yellow oil, which was used further without purification.

[0653] TLC: PE / EA=4:1, KMnO4, ninhydrin

[0654] R f (Compound 2m) = 0.2

[0655] R f (Compound 3m) = 0.4

[0656] LC-MS: none found.

[0657] General procedure for the preparation of compound 5m

[0658]

[0659] To a solution of compound 3m (7.8 g, 41.7 mmol, 1.8 equiv) and compound 4a (6.0 g, 23.1 mmol, 1.0 equiv) in DCE / DMF (1 / 1, 120 mL) was added acetic acid (30 mL). The reaction was stirred at room temperature for 2 h. At this stage, NaBH3CN (5.8 g, 92.6 mmol, 4.0 equiv) was added portionwise at 0°C and stirred at room temperature for 18 h. After the reaction was complete, the reaction mixture was diluted with DCM (600 mL). The mixture was washed with water (2 x 60 mL) and brine (2 x 60 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (DCM / EA: 3 / 1) to obtain compound 5m (2.0 g, 19%) as a white solid.

[0660] TLC: EA

[0661] R f (Compound 3m) = 0.4

[0662] R f (Compound 5m) = 0.6

[0663] LC-MS:445.35[M+1] + ,443.30[M-1] -

[0664] General procedure for the preparation of compound 6m

[0665]

[0666] To a solution of compound 5m (2.0 g, 4.5 mmol, 1.0 equivalent) and compound 6 (2.6 g, 45 mmol, 10.0 equivalent) in DCE / DMF (40 mL, 1 / 1) was added acetic acid (10 mL) and stirred at room temperature for 1 h under a nitrogen atmosphere. NaBH3CN (1.1 g, 18 mmol, 4.0 equivalent) was added in portions and stirred at room temperature for 18 h under a nitrogen atmosphere. After completion of the reaction (confirmed by TLC analysis (100% EA) and LCMS), the reaction mixture was diluted with DCM (200 mL). The mixture was washed with water (2 × 40 mL) and brine (2 × 40 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (eluting with DCM / EA: 4:1) and P-HPLC (acetonitrile / H2O: 40%-60%) to obtain compound 6m (0.87 g, 39%) as a white solid.

[0667] TLC: EA=100%

[0668] R f (Compound 5m) = 0.5

[0669] R f (Compound 6m) = 0.65

[0670] LC-MS:487.40[M+1] + ,485.40[M-1] -

[0671] General procedure for the preparation of compound 57

[0672]

[0673] To a solution of compound 6m (870 mg, 1.8 mmol, 1.0 equiv) in methanol (4 mL) was added HCl / EA (9 mL, 2 N, 18 mmol, 10.0 equiv). The resulting solution was stirred at 30°C for 18 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by P-HPLC (acetonitrile / HO: 30%-40%) to afford compound 57 (144 mg, 30%) as a white solid.

[0674] LC-MS: 387.40[M+1] + ,385.15[M-1] -

[0675] 1H NMR (400MHz, CD3OD): δ7.59(s,3H),5.18(d,J=10.0Hz,1H),4.57(s,2H),3.47(s,3H),3.00–2.74(m,5H ),2.55(d,J=10.6Hz,1H),2.21(s,1H),1.73–1.45(m,7H),1.39(d,J=6.4Hz,3H),0.90(t,J=6.3Hz,4H).

[0676] Compound 58 in the form of hydrochloride

[0677]

[0678] To a solution of compound 61 (0.2 g, 0.5 mmol, 1.0 eq) in THF (2 mL) was added HCl / EA (2.4 mL, 2 N, 4.8 mmol, 10.0 eq) and stirred for 2 days at 30° C. The resulting mixture was concentrated under reduced pressure and purified by P-HPLC (acetonitrile / HO: 30%-40%) to afford compound 58 (102 mg, 52%) as a white solid.

[0679] LC-MS:402.30[M+1] + ,400.20[M-1] -

[0680] 1 H NMR (400MHz, DMSO-d6): δ10.99(s,1H),7.25(s,1H),6.90(d,J=4.0Hz,1H),6.73(d,J=4.0Hz,1H),5.57(s,1H),5.59(d,J=12.0Hz,1 H),4.26-3.85(m,2H),3.05(s,2H),3.85-3.75(m,1H),2.70-2.50(m,1H),2.31-2.20(m,1H),1.99(s,1H),1.55(s,2H),0.89(s,3H).

[0681] The hydrochloride form of compound 60

[0682]

[0683] General procedure for the preparation of compound 2p

[0684]

[0685] Under a nitrogen atmosphere, at -78 ° C, over a period of 15 minutes, a solution of DMSO (4.2 mL, 1.0 V) in DCM (20 mL) was added dropwise to a solution of oxalyl chloride (2.9 g, 22.85 mmol, 1.2 equivalents) at DCM (10 mL), and stirred at the same temperature for 15 minutes. A solution of compound 1p (4.2 g, 19.32 mmol, 1.0 equivalents) in DCM (10 mL) was added dropwise over a period of 15 minutes. After the reaction mixture was stirred at -78 ° C for 30 minutes, TEA (5.8 g, 57.98 mmol, 3.0 equivalents) was added dropwise at -78 ° C. Once the addition was complete, the reaction mixture was slowly warmed to 0 ° C and stirred for 30 minutes. The reaction was complete (analyzed by TLC (PE / EA=4 / 1, R f After 4% HCl (5% NaHCO₄ or ninhydrin) was added, the reaction mixture was diluted with DCM (50 mL). The mixture was washed with 10% citric acid (30 mL), water (50 mL) and brine (50 mL), dried over Na₂SO₄, filtered and concentrated under vacuum below 30° C. to afford compound 2p (4.1 g, crude product) as a yellow oil, which was used without further purification.

[0686] TLC: PE / EA=2:1, KMnO4, ninhydrin

[0687] R f (Compound 1p) = 0.2

[0688] R f (Compound 2p) = 0.6

[0689] LC-MS: none found.

[0690] 1 H NMR (400MHz, DMSO-d6): δ9.63(s,1H),6.75(s,1H),2.86(d,J=5.2Hz,2H),2.38(s,2H),1.56–1.42(m,2H),1.34(s,11H),1.20(d,J=5.4Hz,3H).

[0691] General procedure for the preparation of compound 5p

[0692]

[0693] To a solution of compound 4a (2.07 g, 4.81 mmol, 1 eq) and compound 2p (4.1 g, 19.32 mmol, 4.0 eq) in DCE / DMF (20 mL) was added acetic acid (2.1 mL) and molecular sieves (4.1 g), and stirred at room temperature under a nitrogen atmosphere for 1 h. NaBH3CN (1.51 g, 24.07 mmol, 5 eq) was added portionwise, and stirred at room temperature under a nitrogen atmosphere for 16 h. After completion of the reaction (confirmed by TLC analysis (100% EA) and LCMS), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3 x 20 mL) and brine (20 mL), and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (DCM / EA: 2 / 1) and P-HPLC (acetonitrile / HO: 40%-60%) to obtain compound 5p (1.0 g, 34%) as a white solid.

[0694] TLC: EA=100%

[0695] R f (Compound 4a) = 0.5

[0696] R f (Compound 5p) = 0.6

[0697] LC-MS: 630.60[M+1] +

[0698] General procedure for the preparation of compound 60

[0699]

[0700] To a solution of compound 5p (200 mg, 0.318 mmol, 1 eq) in methanol (1 mL) was added HCl (1.6 mL, 2 N, 3.2 mmol, 10 eq). The resulting solution was stirred at room temperature for 24 h. The reaction mixture was lyophilized to afford compound 60 (130 mg, 76%) as a white solid.

[0701] TLC: DCM / methanol = 10 / 1

[0702] R f (Compound 5p) = 0.6

[0703] R f (Compound 60) = 0.1

[0704] LC-MS:430.40[M+1] +

[0705] 1H NMR (400MHz, D2O): δ7.89(d,J=7.1Hz,1H),7.80–7.67(m,2H),5.13–5.01(m,1H),4.53(t,J=13.3Hz,2H),3 .56(s,4H),2.72(d,J=6.9Hz,6H),2.40(d,J=7.9Hz,1H),2.16(s,1H),1.38(t,J=36.0Hz,8H),1.12(s,4H).

[0706] Compound 61

[0707]

[0708] General procedure for the preparation of compound 2q

[0709]

[0710] To a solution of compound 1q (5.0 g, 50 mmol, 1.0 equivalent) in MeOH (100 mL) was added concentrated sulfuric acid (0.58 mL) and stirred at 90 ° C for 18 h. The solution was cooled to 0 ° C, treated with NaHCO (464 mg), and stirred for 10 minutes. The reaction mixture was filtered and the solvent was removed in vacuo. The residue was diluted with DCM (50 mL), dried over Na SO, filtered and concentrated to obtain compound 2q (5.9 g) as a colorless oil, which was used without further purification.

[0711] TLC: PE / EA=4:1, KMnO4, ninhydrin

[0712] R f (Compound 1q) = 0.4

[0713] R f (Compound 2q) = 0.2

[0714] LC-MS: none found.

[0715] General procedure for the preparation of compound 3q

[0716]

[0717] Under a nitrogen atmosphere, at -78 ° C, over a period of 15 minutes, a solution of oxalyl chloride (7.6 g, 59.9 mmol, 1.2 equivalents) in DCM (33 mL) was added dropwise to a solution of DMSO (6.6 mL) in DCM (66 mL), and stirred at the same temperature for 15 minutes. A solution of compound 2q (6.6 g, 49.9 mmol, 1.0 equivalents) in DCM (33 mL) was added dropwise over a period of 15 minutes. After the reaction mixture was stirred at -78 ° C for 30 minutes, TEA (15.2 g, 149.8 mmol, 3.0 equivalents) was added dropwise at -78 ° C. Once the addition was complete, the reaction mixture was slowly warmed to 0 ° C and stirred for 30 minutes. The reaction was complete (analyzed by TLC (PE / EA=4 / 1, R f After 4% HCl (0.5% NaHCO₄ or ninhydrin) was added, the reaction mixture was diluted with DCM (10 mL). The mixture was washed with 10% citric acid (10 mL), water (10 mL) and brine (10 mL), dried over Na₂SO₄, filtered and concentrated under vacuum below 30° C. to afford compound 3q (7.45 g, crude product) as a yellow oil, which was used without further purification.

[0718] TLC: PE / EA=4:1, KMnO4, ninhydrin

[0719] R f (Compound 2q) = 0.2

[0720] R f (Compound 3q) = 0.4

[0721] LC-MS: none found.

[0722] 1 H NMR (400MHz, CDCl3): δ3.65–3.52(m,5H),2.29(td,J=7.2,3.8Hz,2H),1.88–1.75(m,2H),1.53(ddt,J=13.4,7.4,3.4Hz,2H).

[0723] General procedure for the preparation of compound 61

[0724]

[0725] To a solution of compound 3q (2.5 g, 19.4 mmol, 1.8 equivalents) and compound 5 (3.2 g, 10.8 mmol, 1.0 equivalents) in DCE / DMF (60 mL, 1 / 1) was added acetic acid (15 mL) and stirred at room temperature for 1 h under a nitrogen atmosphere. NaBH3CN (2.7 g, 43 mmol, 4.0 equivalents) was added in portions and stirred at room temperature under a nitrogen atmosphere overnight. After completion of the reaction (confirmed by TLC analysis (100% EA) and LCMS), the reaction mixture was diluted with DCM (400 mL). The mixture was washed with water (2 × 40 mL) and brine (2 × 40 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (DCM / EA: 2 / 1) and by P-HPLC (acetonitrile / H2O: 40%-60%) to obtain compound 61 (0.4 g, 13%) as a white solid.

[0726] TLC: EA=100%

[0727] R f (Compound 5) = 0.5

[0728] R f (Compound 61) = 0.6

[0729] LC-MS:416.35[M+1] +

[0730] 1 H NMR (400MHz, CD3OD): δ7.36(dd,J=25.2,7.3Hz,2H),7.17(d,J=7.8Hz,1H),5.19–5.08(m,1H),4.47(s,2H),3.60(s,3H),3.23–3.11(m,4H),2.83(d d,J=43.9,15.0Hz,2H),2.57(d,J=13.6Hz,1H),2.31(t,J=6.6Hz,2H),2. 18(d,J=12.4Hz,1H), 1.55(dd,J=34.0,6.5Hz,6H), 0.88(t,J=6.9Hz,3H).

[0731] Compound 63

[0732]

[0733] To a solution of compound 58 (120 mg, 0.3 mmol, 1.0 eq) in THF (2 mL) was added HATU (136 mg, 0.4 mmol, 1.2 eq), DIEA (151 mg, 1.5 mmol, 5.0 eq) and MeNH2.HCl (22.3 mg, 0.33 mmol, 1.1 eq). The resulting solution was stirred at room temperature for 18 h. The resulting mixture was concentrated under reduced pressure and purified by P-HPLC (acetonitrile / H2O: 30%-40%) to obtain compound 63 (63.4 mg, 50%) as a white solid.

[0734] LC-MS:415.35[M+1] + ,413.25[M-1] -

[0735] 1 H NMR (400MHz, CD3OD): δ7.82(s,1H),7.36(dd,J=25.5,6.7Hz,2H),7.18(d,J=7.4Hz,1H),5.13(d,J=12.6Hz,1H),4.46(s,2H),3.18(d,J=23.8Hz,5H),2 .83(dd,J=40.3,16.7Hz,2H),2.65(s,3H),2.56(d,J=13.2Hz,1H),2.14(s, 3H), 1.55 (d, J = 37.7Hz, 6H), 1.32 (d, J = 26.3Hz, 3H), 0.89 (d, J = 6.5Hz, 3H).

[0736] Compound 66 in the form of hydrochloride

[0737]

[0738] General procedure for the preparation of compound 2

[0739]

[0740] To a solution of compound 1 (0.5 g, 8.77 mmol, 1.0 equivalent) in DCM (10 mL) was added TEA (2.7 g, 26.3 mmol, 3.0 equivalents) and 9-fluorenylmethyl chloroformate (3.4 g, 13.2 mmol, 1.0 equivalents). The mixture was stirred at room temperature for 16 h under a nitrogen atmosphere. After completion of the reaction (confirmed by TLC analysis), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3 × 20 mL) and brine (20 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA: 10 / 1) to obtain compound 2 (0.4 g, 16%) as a white solid.

[0741] TLC: PE / EA=1 / 1, ninhydrin

[0742] R f (Compound 1) = 0.1

[0743] R f (Compound 2) = 0.8

[0744] 1 H NMR (400MHz, CDCl3): δ7.76(d,J=6.4Hz,2H),7.59(d,J=6.9Hz,2H),7.45-7.39(m,2H),7.39-7.27( m,2H),5.84(brs,1H),5.25-5.05(m,2H),4.82(s,1H),4.45-4.30(m,2H),4.22(s,1H),3.82(s,2H).

[0745] General procedure for the preparation of compound 3

[0746]

[0747] To a solution of compound 2 (0.3 g, 1.1 mmol, 1.0 eq) in DCM (10 mL) were added acrolein (69 mg, 1.2 mmol, 1.1 eq) and HG 2nd (34 mg, 0.05 mmol, 0.05 equiv). The mixture was stirred at 50 ° C overnight under a nitrogen atmosphere. After the reaction was completed (confirmed by TLC analysis), the reaction mixture was diluted with DCM (10 mL). The mixture was washed with water (10 mL), brine (10 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA: 4 / 1) to obtain compound 3 (0.28 g, 85%) as a yellow solid.

[0748] TLC: PE / EA=3 / 1

[0749] R f (Compound 2) = 0.4

[0750] R f (Compound 3) = 0.2

[0751] 1 H NMR (400MHz, CDCl3): δ9.56 (d, J = 6.1Hz, 1H), 7.75 (s, 2H), 7.57 (s, 2H), 7.47-7.25 (m, 4H), 6. 77(d,J=14.8Hz,1H),6.25-6.10(m,1H),5.03(s,1H),4.47(s,2H),4.21(s,1H),4.07(s,2H).

[0752] General procedure for the preparation of compound 5

[0753]

[0754] To a solution of compound 4 (450 mg, 1.7 mmol, 1.0 equiv) and compound 3 (1.1 g, 3.5 mmol, 2.0 equiv) in DCE / DMF (3 / 1, 8 mL) was added acetic acid (1.2 mL) and molecular sieves (1.0 g). The mixture was stirred at room temperature under a nitrogen atmosphere for 1 h, and then NaBH3CN (437 mg, 6.9 mmol, 4.0 equiv) was added portionwise. The reaction mixture was stirred at room temperature under a nitrogen atmosphere overnight. After completion of the reaction (confirmed by TLC analysis and LCMS), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3×20 mL), brine (20 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (DCM / EA: 2 / 1) to obtain compound 5 (157 mg, 19%) as a white solid.

[0755] TLC: EA=100%

[0756] R f (Compound 4) = 0.4

[0757] R f (Compound 5) = 0.5

[0758] LCMS: [C 32 H 30 N4O5] + Calculated value: 551, found value: 551.

[0759] General procedure for the preparation of compound 6

[0760]

[0761] To a solution of compound 5 (137 mg, 0.25 mmol, 1.0 equiv) in DCE / DMF (3 / 1, 8 mL) was added butyraldehyde (71.8 mg, 1.0 mmol, 4.0 equiv), acetic acid (1.0 mL) and molecular sieves (1.0 g). The mixture was stirred at room temperature under a nitrogen atmosphere for 1 h, and NaBH3CN (62.6 mg, 1.0 mmol, 4.0 equiv) was added portionwise. The reaction mixture was stirred at room temperature under a nitrogen atmosphere overnight. After completion of the reaction (confirmed by TLC analysis and LCMS), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3×20 mL), brine (20 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA: 1 / 1) to obtain compound 6 (41 mg, 27%) as a white solid.

[0762] TLC: EA=100%

[0763] R f (Compound 5) = 0.5

[0764] R f (Compound 6) = 0.6

[0765] LCMS: [C 36 H 38 N4O5] + Calculated value: 607, found value: 607.

[0766] General procedure for the preparation of compound 66

[0767]

[0768] To a solution of compound 6 (47 mg, 0.1 mmol, 1.0 equiv) in DCM (4.0 mL) was added piperidine (66.7 mg, 0.8 mmol, 8.0 equiv). The mixture was stirred at room temperature under a nitrogen atmosphere overnight. The reaction mixture was concentrated under reduced pressure and purified by P-HPLC (acetonitrile / HO: 20%-40%) to afford compound 66 (4.4 mg, 13%) as a white solid.

[0769] TLC: DCM / methanol = 10 / 1

[0770] R f (Compound 6) = 0.8

[0771] R f (Compound 66) = 0.1

[0772] LCMS: [C 21 H 28 N4O3] + Calculated value: 385, found value: 385.

[0773] 1 H NMR (400MHz, D2O): δ7.79(d,J=6.3Hz,1H),7.75-7.55(m,2H),5.80-5.70(m,1H),5.70-5.55(m,1H),5.07(d,J=12.5Hz,1H),4.55-4.42(m,2H) ,4.20-4.05(m,2H),3.61-3.37(m,2H),3.34(s,2H),2.87-2.68(m,1H) ,2.50-2.05(m,1H),1.37-1.24(m,2H),1.24-1.05(m,2H),0.67(s,3H).

[0774] Compound 67 in the form of hydrochloride

[0775]

[0776] General procedure for the preparation of compound 2

[0777]

[0778] To a solution of compound 1 (3.0 g, 34.8 mmol, 1.0 equiv) and tert-butylchlorodiphenylsilane (2.9 g, 10.5 mmol, 0.3 equiv) in THF (60 mL) was added TEA (17.6 g, 174.3 mmol, 5.0 equiv), DMAP (0.4 g, 3.5 mmol, 0.1 equiv). The mixture was stirred at room temperature for 17 hours under a nitrogen atmosphere. After completion of the reaction (confirmed by TLC analysis), the reaction mixture was diluted with DCM (50 mL). The mixture was washed with water (3 × 20 mL), brine (20 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA: 10 / 1) to obtain compound 2 (3.0 g, 88%) as a clear oil.

[0779] TLC: PE / EA=5 / 1

[0780] R f (Compound 1) = 0.1

[0781] R f (Compound 2) = 0.3

[0782] 1H NMR (400MHz, CDCl3): δ7.75-7.65(m,4H),7.50-7.77(m,6H),4.35(s,2H),4.19(s,2H),1.05(s,9H).

[0783] General procedure for the preparation of compound 3

[0784]

[0785] Under nitrogen atmosphere, at 0 ℃, over 15 minutes, to a solution of compound 2 (3.0 g, 9.3 mmol, 1.0 equivalent) in DCM (30 mL) was added a solution of TEA (2.8 g, 27.8 mmol, 3.0 equivalent) and MsCl (1.59 g, 13.9 mmol, 1.5 equivalent) in DCM (35 mL). The mixture was stirred at the same temperature for 15 minutes. The reaction mixture was then slowly warmed to room temperature and stirred for 4 hours. After the reaction was complete (confirmed by TLC analysis), the reaction mixture was diluted with DCM (50 mL) and washed with water (50 mL) and brine (50 mL). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE / EA: 10 / 1) to obtain compound 3 (2.9 g, 78%) as a clear oil.

[0786] TLC: PE / EA=2 / 1

[0787] R f (Compound 2) = 0.60

[0788] R f (Compound 3) = 0.63

[0789] LCMS: [C 21 H 26 O4SSi] + Calculated value of Na: 425, found: 425.

[0790] 1 H NMR (400MHz, CDCl3): δ7.75-7.65(m,4H),7.50-7.77(m,6H),4.81(s,2H),4.37(s,2H),3.02(s,3H),1.04(s,9H).

[0791] General procedure for the preparation of compound 4

[0792]

[0793] Compound 3 (2.9 g, 7.2 mmol, 1 equivalent) was dissolved in NH / MeOH (29 mL). The mixture was stirred at room temperature for 17 hours. After the reaction was complete (confirmed by TLC analysis and LCMS), the reaction mixture was concentrated under reduced pressure and purified by silica gel column chromatography (DCM / MeOH: 20 / 1) to obtain compound 4 (1.9 g, 82%) as a white solid.

[0794] TLC: DCM / EA=1 / 1

[0795] R f (Compound 3) = 0.9

[0796] R f (Compound 4) = 0.1

[0797] LCMS: [C 20 H 25 NOSi] + Calculated value: 324, found value: 324.

[0798] 1 H NMR (400MHz, CDCl3): δ8.26(s,1H),7.75-7.65(m,4H),7.50-7.77(m,6H),4.29(s,2H),3.52(s,2H),1.03(s,9H).

[0799] General procedure for the preparation of compound 5

[0800]

[0801] To a solution of compound 4 (2.08 g, 6.4 mmol, 1 eq) in DCM (20 mL) was added Fmoc-Osu (2.6 g, 7.7 mmol, 1.2 eq) and TEA (1.94 g, 19.3 mmol, 3.0 eq). The mixture was stirred at room temperature under a nitrogen atmosphere for 17 hours. After completion of the reaction (confirmed by TLC analysis and LCMS), the reaction mixture was diluted with DCM (20 mL) and washed with water (3×20 mL) and brine (20 mL). The organic layer was dried over Na 2 SO 4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE / EA: 1 / 1) to obtain compound 5 (2.6 g, 74%) as a white solid.

[0802] TLC: DCM / EA=1 / 1

[0803] R f (Compound 4) = 0.1

[0804] R f(Compound 5) = 0.8

[0805] LCMS: [C 35 H 35 NO3Si] + Calculated value of Na: 568, found: 568.

[0806] General procedure for the preparation of compound 6

[0807]

[0808] To a solution of compound 5 (2.6 g, 4.8 mmol, 1.0 equiv) in THF (1.0 mL) was added TEA-3HF (7.7 g, 47.7 mmol, 10.0 equiv). The resulting solution was stirred at room temperature overnight. After completion of the reaction (confirmed by TLC analysis and LCMS), the reaction was quenched with H2O (20 mL). The reaction mixture was extracted with EA (20 mL × 3). The organic layer was washed with water (3 × 20 mL), brine (20 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA: 3 / 1) to obtain compound 6 (1.0 g, 68%) as a white solid.

[0809] TLC: PE / EA=2 / 1

[0810] R f (Compound 5) = 0.8

[0811] R f (Compound 6) = 0.2

[0812] 1 H NMR (400MHz, CDCl3): δ7.76(d,J=7.5Hz,2H),7.58(d,J=7.0Hz,2H),7.40(t,J=7.4Hz,2 H),7.31(t,J=7.4Hz,2H),4.43(d,J=6.8Hz,2H),4.27(s,2H),4.22(s,1H),4.03(s,2H).

[0813] General procedure for the preparation of compound 7

[0814]

[0815] To a solution of compound 6 (1.5 g, 4.9 mmol, 1.0 equiv) in DCM (20 mL) was added MnO2 (4.24 g, 49.0 mmol, 10.0 equiv). The mixture was stirred at room temperature overnight. The reaction mixture was filtered and concentrated to afford compound 7 (1.15 g, 77%) as a white solid. It was used directly in the next step without further purification.

[0816] TLC: PE / EA=2 / 1

[0817] R f (Compound 6) = 0.2

[0818] R f (Compound 7) = 0.5

[0819] 1 H NMR (399MHz, CDCl3): δ9.18(s,1H),7.75(d,J=7.5Hz,2H),7.57(d,J=7.4Hz,2H),7.39(t,J =7.4Hz,2H),7.30(t,J=7.4Hz,2H),5.02(s,1H),4.45(d,J=6.6Hz,2H),4.27-4.10(m,2H).

[0820] General procedure for the preparation of compound 9

[0821]

[0822] To a solution of compound 8 (1.0 g, 3.8 mmol, 1.0 equivalent) and compound 7 (1.2 g, 3.8 mmol, 1.0 equivalent) in DCE / DMF (6 mL) was added acetic acid (1.5 mL) and molecular sieves (1.76 g). The mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. NaBH3CN (1.0 g, 15.3 mmol, 4.0 equivalents) was then added portionwise and stirred at room temperature for another 16 hours under a nitrogen atmosphere. After completion of the reaction (confirmed by TLC analysis and LCMS), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3 × 20 mL), brine (20 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by P-HPLC (acetonitrile / H2O: 40%-60%) to obtain compound 9 (1.3 g, 63%) as a white solid.

[0823] TLC: EA=100%

[0824] R f (Compound 8) = 0.4

[0825] Rf (Compound 9) = 0.5

[0826] General procedure for the preparation of compound 10

[0827]

[0828] To a solution of compound 9 (1.2 g, 2.2 mmol, 1.0 equiv) and butyraldehyde (0.6 g, 8.7 mmol, 4.0 equiv) in DCE / DMF (3 / 1, 4 mL) was added acetic acid (1.5 mL) and molecular sieves (1.8 g). The mixture was stirred at room temperature under a nitrogen atmosphere for 1 hour. NaBH3CN (546 mg, 8.7 mmol, 4.0 equiv) was then added portionwise and stirred at room temperature under a nitrogen atmosphere for an additional 16 hours. After completion of the reaction (confirmed by TLC analysis and LCMS), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3 x 20 mL), brine (20 mL) and dried over Na2SO4. The solvent was removed under reduced pressure and the residue was purified by P-HPLC (acetonitrile / H2O: 40%-60%) to obtain compound 10 (315 mg, 24%) as a white solid.

[0829] TLC: EA=100%

[0830] R f (Compound 9) = 0.5

[0831] R f (Compound 10) = 0.6

[0832] LCMS: [C 36 H 36 N4O5] + Calculated value: 605, found value: 605.

[0833] General procedure for the preparation of compound 11

[0834]

[0835] To a solution of compound 10 (587 mg, 1.0 mmol, 1.0 eq) in EtOH (5 mL) was added Lindela's catalyst (60 mg, 10%) and quinoline (12.5 mg, 0.1 mmol, 0.1 eq). Under N2 / H2 (50%), the mixture was stirred at room temperature for 30 minutes. The reaction mixture was filtered, concentrated under reduced pressure and purified by P-HPLC (acetonitrile / H2O: 40%-60%) to obtain compound 11 (100 mg, 17%) as a white solid.

[0836] TLC: DCM / EA=1 / 1

[0837] R f (Compound 10) = 0.7

[0838] R f (Compound 11) = 0.72

[0839] LCMS: [C 36 H 38 N4O5] + Calculated value: 607, found value: 607.

[0840] General procedure for the preparation of compound 67

[0841]

[0842] To a solution of compound 11 (100 mg, 0.17 mmol, 1.0 equiv) in DCM (5.0 mL) was added piperidine (280 mg, 3.3 mmol, 20.0 equiv). The mixture was stirred at room temperature for 17 hours. The reaction mixture was then concentrated under reduced pressure and purified by P-HPLC (acetonitrile / HO: 20%-45%) to afford compound 67 (14.2 mg, 19%) as a white solid.

[0843] TLC: DCM / methanol = 10 / 1

[0844] R f (Compound 11) = 0.9

[0845] R f (Compound 67) = 0.1

[0846] LCMS: [C 21 H 28 N4O3] + Calculated value: 385, found value: 385.

[0847] 1H NMR (400MHz, D2O): δ7.84(dd,J=15.1,7.8Hz,2H),7.69(t,J=7.8Hz,1H),5.72-5.62(m,1 H),5.62-5.50(m,1H),5.05(dd,J=13.3,5.2Hz,1H),4.51(q,J=17.5Hz,2H),4.23(d,J=7 .4Hz,2H),3.66-3.52(m,2H),3.38-3.21(m,2H),2.85-2.65(m,2H),2.39(td,J=12.9,7. 7Hz,1H),2.22-2.05(m,1H),1.35-1.20(m,2H),1.19-1.06(m,2H),0.63(t,J=7.3Hz,3H).

[0848] Compound 68 in the form of hydrochloride

[0849]

[0850] General procedure for the preparation of compound 2

[0851]

[0852] To a solution of compound 1 (1.0 g, 5.28 mmol, 1.0 eq) in EA (3 mL) was added HCl / EA (17.6 mL, 3 N, 52.8 mmol, 10.0 eq) and stirred at room temperature for 3 h under a nitrogen atmosphere. After completion of the reaction (confirmed by TLC analysis), the solution was concentrated in vacuo to afford compound 2 (1.2 g, crude product) as a yellow oil, which was used directly without further purification.

[0853] TLC: PE / EA=1 / 1KMnO4

[0854] R f (Compound 1) = 0.4

[0855] R f (Compound 2) = 0.1

[0856] General procedure for the preparation of compound 3

[0857]

[0858] To a solution of compound 2 (0.3 g, 3.4 mmol, 1.0 eq) in THF / HO (3 / 1, 8 mL) was added Fmoc-OSu (1.7 g, 5.1 mmol, 1.5 eq) and TEA (340 mg, 3.4 mmol, 1.0 eq). The mixture was stirred at room temperature overnight. After completion of the reaction (confirmed by TLC analysis), the reaction mixture was diluted with DCM (10 mL). The mixture was washed with water (10 mL), brine (5 mL) and dried over NaSO. The resulting mixture was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA: 5 / 1) to obtain compound 3 (160 mg, 21%) as a white solid.

[0859] TLC: PE / EA=2 / 1

[0860] R f (Compound 2) = 0.1

[0861] R f (Compound 3) = 0.7

[0862] 1 H NMR (400MHz, CDCl3): δ7.75(d,J=7.5Hz,2H),7.57(d,J=7.4Hz,2H),7.38(t,J=7.3Hz,2H),7.30( t,J=8.5,2H),4.50-4.4(m,2H),4.25-4.20(m,1H),3.65(s,2H),3.21(s,2H),1.75-1.37(m,4H).

[0863] General procedure for the preparation of compound 4

[0864]

[0865] Under nitrogen atmosphere, at -78 ℃, over 15 minutes, to a solution of oxalyl chloride (200mg, 1.6mmol, 1.2 equivalents) in DCM (3mL), a solution of DMSO (0.4mL, 1.0V) in DCM (2mL) was added dropwise, and stirred at the same temperature for 15 minutes. At -78 ℃, a solution of compound 3 (410mg, 1.32mmol, 1.0 equivalents) in DCM (3mL) was added to the previous solution over 15 minutes. After the mixture was stirred at -78 ℃ for 30 minutes, TEA (400mg, 3.96mmol, 3.0 equivalents) was added at -78 ℃. Once the addition was complete, the reaction mixture was slowly warmed to 0 ℃ and stirred for 30 minutes. After the reaction was completed (confirmed by TLC analysis), the reaction mixture was diluted with DCM (20mL) and quenched with 10% citric acid aqueous solution (10mL). The resulting solution was washed with water (20 mL), brine (20 mL) and dried over Na 2 SO 4 . The solvent was removed under vacuum below 30° C. to afford compound 4 (640 mg, crude product) as a yellow oil, which was used directly without further purification.

[0866] TLC: PE / EA=4:1, KMnO4, ninhydrin

[0867] R f (Compound 3) = 0.4

[0868] R f (Compound 4) = 0.6

[0869] General procedure for the preparation of compound 6

[0870]

[0871] To a solution of compound 5 (410 mg, 1.6 mmol, 1.0 equivalent) and compound 4 (979 mg, 3.2 mmol, 2.0 equivalent) in DCE / DMF (10 mL) was added acetic acid (1.5 mL) and molecular sieves (1.0 g). The mixture was stirred at room temperature under a nitrogen atmosphere for 1 hour. NaBH3CN (398 mg, 6.4 mmol, 4.0 equivalent) was then added portionwise and stirred at room temperature under a nitrogen atmosphere overnight. After the reaction was complete (confirmed by TLC analysis), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3 × 30 mL), brine (30 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by P-HPLC (acetonitrile / H2O: 40%-60%) to obtain compound 6 (315 mg, 35%) as a white solid.

[0872] TLC: EA=100%

[0873] R f (Compound 5) = 0.4

[0874] R f (Compound 6) = 0.5

[0875] General procedure for the preparation of compound 7

[0876]

[0877] To a solution of compound 6 (642 mg, 1.2 mmol, 1.0 equivalent) in THF (3.0 mL) was added dropwise a solution of crotonaldehyde (87.6 mg, 1.3 mmol, 1.1 equivalent) in H2SO4 (3.11 mL, 4N, 12.4 mmol, 10.7 equivalent) at 0 ° C. Then NaBH4 (26.8 mg, 0.7 mmol, 0.6 equivalent) was added at 0 ° C. and stirred at the same temperature for 40 minutes. The reaction mixture was poured into ice water and extracted with DCM (3 × 30 mL). The organic layer was washed with H2O, brine and concentrated under reduced pressure. The residue was purified by P-HPLC (acetonitrile / H2O: 40%-60%) to obtain compound 7 (120 mg, 17%) as a white solid.

[0878] TLC: EA=100%

[0879] R f (Compound 6) = 0.5

[0880] R f (Compound 7) = 0.6

[0881] LCMS: [C 36 H 38 N4O5] + Calculated value: 607, found value: 607.

[0882] General procedure for the preparation of compound 68

[0883]

[0884] To a solution of compound 7 (120 mg, 0.2 mmol, 1.0 eq) in DCM (4.0 mL) was added piperidine (171 mg, 2.0 mmol, 10.0 eq). The mixture was stirred at room temperature for 17 hours. After vacuum concentration, the residue was purified by P-HPLC (acetonitrile / HO: 20%-40%) to obtain compound 68 (42.5 mg, 46%) as a white solid.

[0885] TLC: DCM / methanol = 10 / 1

[0886] R f (Compound 7) = 0.5

[0887] R f (Compound 68) = 0.1

[0888] LCMS: [C 21 H 28 N4O3] + Calculated value: 385, found value: 385.

[0889] 1 H NMR (400MHz, D2O): δ7.76 (d, J = 7.4Hz, 1H), 7.69 (d, J = 8.1Hz, 1H), 7.60 (t, J = 7. 8Hz,1H),5.75-5.55(m,1H),5.16-5.04(m,1H),4.98(dd,J=13.3,5.1Hz,1H),4 .42(dd,J=35.2,17.5Hz,2H),3.96(d,J=7.2Hz,2H),3.62-3.41(m,2H),2.75-2 .55(m,4H),2.32(qd,J=13.0,5.1Hz,1H),2.10-2.01(m,1H),1.51-1.24(m,7H).

[0890] Compound 69 in the form of hydrochloride salt

[0891]

[0892] General procedure for the preparation of compound 2

[0893]

[0894] Under nitrogen atmosphere, to compound 1 (620mg, 8.8mmol, 1.0 equivalent) in DCM (30mL) solution was added MnO2 (7.7g, 88.5mmol, 10.0 equivalent), molecular sieves (3.1g), and stirred at room temperature for 17 hours. After the reaction was complete (confirmed by TLC analysis), the reaction mixture was filtered and concentrated under reduced pressure below 30°C to obtain compound 2 (110mg, 17%) as a clear oil, which was used directly without further purification.

[0895] TLC: PE / EA=3 / 1KMnO4

[0896] R f (Compound 1) = 0.3

[0897] R f(Compound 2) = 0.3

[0898] 1 H NMR (400MHz, CDCl3): δ9.13 (s, 1H), 2.05 (s, 3H).

[0899] General procedure for the preparation of compound 4

[0900]

[0901] To a solution of compound 3 (2.0 g, 4.6 mmol, 1.0 equivalent) and compound 2 (1.62 g, 18.4 mmol, 4.0 equivalents) in DCE / DMF (3 / 1, 8 mL) was added acetic acid (2.0 mL) and molecular sieves (4.0 g). The mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. NaBH3CN (1.2 g, 18.4 mmol, 4.0 equivalents) was then added portionwise and stirred at room temperature under a nitrogen atmosphere overnight. After completion of the reaction (confirmed by TLC analysis and LCMS), the reaction mixture was diluted with DCM (20 mL). The mixture was washed with water (3 × 20 mL), brine (20 mL) and dried over Na2SO4. The resulting mixture was concentrated under reduced pressure and purified by P-HPLC (acetonitrile / H2O: 40%-60%) to obtain compound 4 (110 mg, 5%) as a white solid.

[0902] TLC: EA=100%

[0903] R f (Compound 3) = 0.4

[0904] R f (Compound 4) = 0.5

[0905] LCMS: [C 26 H 34 N4O5] + Calculated value: 483, found value: 483.

[0906] General procedure for the preparation of compound 5

[0907]

[0908] To a solution of compound 4 (100 mg, 0.2 mmol, 1.0 eq) in EtOH (5 mL) was added Lindela's catalyst (10 mg, 10%) and quinoline (1.8 mg, 0.02 mmol, 0.1 eq). The mixture was stirred at room temperature for 30 minutes, filtered and concentrated under reduced pressure. The residue was purified by P-HPLC (acetonitrile / HO: 40%-60%) to obtain compound 5 (20 mg, 20%) as a white solid.

[0909] TLC: DCM / methanol = 1 / 1

[0910] R f (Compound 4) = 0.6

[0911] R f (Compound 5) = 0.62

[0912] LCMS: [C 26 H 36 N4O5] + Calculated value: 485, found value: 485.

[0913] General procedure for the preparation of compound 69

[0914]

[0915] To a solution of compound 5 (33 mg, 0.07 mmol, 1.0 eq) in MeOH (1 mL) was added HCl (0.4 mL, 2 N, 0.8 mmol, 11.0 eq). The mixture was stirred at room temperature for 24 hours and concentrated under reduced pressure. The residue was purified by Biotage (C18, acetonitrile / HO: 30%-60%) to afford compound 69 (17.8 mg, 57%) as a white solid.

[0916] TLC: DCM / methanol = 10 / 1

[0917] R f (Compound 5) = 0.5

[0918] R f (Compound 69) = 0.1

[0919] LCMS: [C 21 H 28 N4O3] + Calculated value: 385, found value: 385.

[0920] 1H NMR (400MHz, D2O): δ7.87-7.77(m,2H),7.72-7.61(m,1H),5.80-5.61(m,1H),5.27-5.12(m,1H),5.04(d,J=10.0Hz,1H),4.50(q,J=17.4Hz,2H ),4.16(d,J=6.2Hz,2H),3.70-3.55(m,2H),2.82-2.61(m,4H),2.48-2. 30(m,1H),2.20-2.04(m,1H),1.58-1.31(m,4H),1.22(d,J=5.2Hz,3H).

[0921] Compound 80 in the form of hydrochloride salt

[0922] 3-(4-((4-(Methylamino)butyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0923]

[0924] Step 1: tert-Butyl methyl(4-oxobutyl)carbamate

[0925]

[0926] Under nitrogen atmosphere, at-78 ℃, within 5 minutes, to the solution of oxalyl chloride (0.25mL, 2.95mmol, 1.2 equivalents) in DCM (5mL), dropwise add DMSO (0.419mL, 5.90mmol, 2.4 equivalents), and stirred 15 minutes at the same temperature.Within 5 minutes, dropwise add solution of (4-hydroxybutyl) (methyl) tert-butyl carbamate 1 (500mg, 5.90mmol, 1 equivalent) in DCM (2.5mL).After stirring reaction mixture 30 minutes at-78 ℃, dropwise add triethylamine (1.713mL, 12.30mmol, 5 equivalents) at-78 ℃.After interpolation, reaction mixture is slowly warmed to 0 ℃ and stirred 30 minutes.Reaction is completed (analyze (50% EtOAc / petroleum ether, R by TLC)). f ~0.4, KMnO4) confirmed), the reaction mixture was diluted with DCM (20 mL) and washed with 10% aqueous citric acid (20 mL), water (1×10 mL) and brine (1×10 mL), dried over anhydrous Na2SO4, filtered and concentrated under vacuum (below 30°C) to afford tert-butyl methyl(4-oxobutyl)carbamate 2 (450 mg, crude product) as a light yellow liquid, which was used further without purification.

[0927] 1 H-NMR (400MHz, DMSO-d6): δ9.67(s,1H),3.16(t,J=7.20Hz,2H),2.69(s,3H),2.43-2.38(m,2H),1.71(t,J=6.80Hz,2H),1.41(s,3H).

[0928] Step 2: tert-Butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)(methyl)carbamate

[0929]

[0930] Under nitrogen atmosphere, a mixture of tert-butyl methyl (4-oxobutyl) carbamate (279 mg, 1.389 mmol, 1.2 equivalents) and lenalidomide 3 (300 mg, 1.157 mmol, 1 equivalent) was dissolved in 1,2-dichloroethane (3 mL, 10 volumes) and DMF (3 mL, 10 volumes). TFA (0.08 mL, 1.041 mmol, 0.9 equivalents) and Na (OAc) 3BH (368 mg, 1.736 mmol, 1.5 equivalents) were added to the solution in portions at 0 ° C., and stirred at room temperature for 16 h. The reaction was completed (by TLC analysis (100% EtOAc, R f After 40 ℃ of drying (10 ℃ of dryings), the reaction mixture was diluted with DCM (20 mL), washed with water (3×20 mL), brine (20 mL), dried over Na 2 SO 4 , filtered and concentrated to afford the crude product, which was purified by Isolera (Biotage R snap cartridge, KP-Sil, 25 g, 230-400 mesh silica gel) using 80-85% ethyl acetate in petroleum ether. Pure fractions were collected and concentrated in vacuo to afford tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)(methyl)carbamate 4 (350 mg, 0.760 mmol, 65% yield) as an off-white solid.

[0931] 1H-NMR (400MHz, DMSO-d6): δ11.00(s,1H),7.29(t,J=7.60Hz,1H),6.93(d,J=6.80Hz,1H),6.75(d,J=8.40Hz,1H),5.61(s,1H),5.12(q,J=5.20Hz, 1H),4.23(d,1H),4.13(d,J=17.20Hz,1H),3.21-3.14(m,3H),2.88(q,2H ),2.72(t,3H),2.58(d,2H),2.46-2.28(m,2H),1.55(s,3H),1.37(s,9H).

[0932] LCMS: 345.1 (M-Boc), Rt (min): 2.267, area %: 96.498.

[0933] Step 3: tert-Butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)(methyl)carbamate

[0934]

[0935] Under nitrogen atmosphere, a mixture of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)(methyl)carbamate 4 (350 mg, 0.787 mmol, 1 eq) and valeraldehyde (339 mg, 3.94 mmol, 5 eq) was dissolved in a mixture of CHCl (3.5 mL, 10 vol) and DMF (3.5 mL, 10 vol). TFA (0.05 mL, 0.709 mmol, 0.9 eq) and STAB (250 mg, 1.181 mmol, 1.5 eq) were added portionwise at 0°C and stirred at room temperature for 16 h. The reaction was complete (by TLC analysis (100% EtOAc, R f0.241mmol, 30% yield).

[0936] 1 H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.38(t,J=8.00Hz,1H),7.19(d,J= 7.20Hz,1H),7.08(d,J=8.40Hz,1H),5.11(q,J=5.20Hz,1H),4.34(d,1H),4. 29(d,1H),3.22-3.11(m,6H),2.92-2.87(m,2H),2.70(q,J=2.00Hz,3H),2.5 7(s,2H),2.01(t,J=5.20Hz,1H),1.25-1.40(m,18H),0.84(t,J=7.20Hz,3H).

[0937] LCMS: 515.4 (M+H), Rt (min): 1.989, Area %: 95.589.

[0938] Step 4: 3-(4-((4-(methylamino)butyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[0939]

[0940] To an ice-cold suspension of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate (130 mg, 0.240 mmol, 1 eq) in DCM (1.5 mL) was added HCl (4 M in EtOAc, 0.7 mL) and stirred at room temperature for 2 h. The reaction progress was monitored by LCMS. The reaction mixture was concentrated, and the residue was triturated with MTBE (10 mL). The obtained solid was lyophilized to afford 3-(4-((4-(methylamino)butyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride 80 (70 mg, 63.2%) as a light yellow solid.

[0941] 1 H-NMR (400MHz, DMSO-d6): δ11.01(s,1H),8.83(s,2H),7.53(s,3H),5.15-5.10(m,1H),4.48-4.49(m,2H),3.31(s,4H),2.97-2.88(m,1H),2. 82(s,1H),2.74(s,2H),2.51-2.48(m,3H),1.91(s,1H),1.59(t,J=6.4 0Hz, 2H), 1.46 (d, J = 17.20Hz, 3H), 1.24 (s, 4H), 0.82 (t, J = 6.80Hz, 3H).

[0942] LCMS: 415.2 (M+H), method: mobile phase: A: 0.1% FA in H2O, mobile phase: B: ACN, column: Atlantis dC18 (50×4.6 mm) 5 μm, flow rate: 1.5 mL / min Rt (min): 1.599, area %: 95.709.

[0943] HPLC: Method: Mobile phase: A: 0.1% TFA in water, Mobile phase: B: ACN, Column: X-Bridge C8 (50×4.6) mm, 3.5 μm, Flow rate: 2.0 mL / min, Rt (min): 2.409, Area %: 97.597.

[0944] Formate salt form of compound 81

[0945] 3-(4-((4-(dimethylamino)butyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione formate

[0946]

[0947] To a solution of 3-(4-((4-aminobutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione 47 (300 mg, 0.749 mmol) and paraformaldehyde (450 mg, 14.98 mmol, 20 eq) in DCM (6 mL, 20 vol) was added AcOH (0.129 ml, 2.247 mmol, 3 eq) and the reaction mixture was stirred at room temperature for 2 h. NaCNBH4 (188 mg, 3.00 mmol, 4 eq) was added portionwise to the mixture and the reaction mixture was stirred at room temperature for another 40 h. The reaction was complete (by TLC analysis (EtOAc, R f After 40 ℃ of drying (confirmed), the reaction mixture was diluted with DCM (10 mL), washed with water (2×20 mL), brine (20 mL), dried over anhydrous Na 2 SO 4 , filtered and concentrated. The resulting residue was purified by preparative HPLC (HCOOH:ACN method) and lyophilized to afford 3-(4-((4-(dimethylamino)butyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione 81 (26 mg, 0.059 mmol, 7.90% yield) as a white solid.

[0948] 1 H-NMR (400MHz, DMSO-d6): δ11.01(s,1H),7.42-7.38(m,1H),7.22(t,J=7.20Hz,1H),7. 12(q,J=3.20Hz,1H),5.23-5.18(m,1H),5.14-5.05(m,1H),4.40(d,J=7.20Hz,1H),4.3 2(d,J=4.40Hz,1H),3.25-3.17(m,4H),2.82-2.68(m,3H),2.53-2.50(m,6H),2.34(s,2 H),1.63(q,J=3.20Hz,2H),1.44(d,J=6.40Hz,4H),1.27-1.23(m,4H),0.85-0.9(m,3H).

[0949] LCMS: 429.3 (M+H). Method: Mobile Phase: A: 0.1% TFA in H2O. Mobile Phase: B: 0.1% TFA in ACN. Flow Rate: 1.5 mL / min. Column: XBridge C8 (50 × 4.6 mm), 3.5 μm. Rt (min): 1.824; Area %: 97.584.

[0950] HPLC: Method: Mobile Phase A: 0.1% TFA in H2O. Mobile Phase B: Acetonitrile. Flow Rate: 2.0 mL / min. Column: X-Bridge C8 (50 x 4.6 mm, 3.5 μm). Rt (min): 3.658; Area %: 97.694.

[0951] Compound 82

[0952] N-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)acetamide

[0953]

[0954] To a solution of 3-(4-((4-aminobutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione 47 (200 mg, 0.499 mmol) in pyridine (3 mL) was added acetic anhydride (0.236 mL, 2.497 mmol, 5 eq) dropwise and the reaction mixture was stirred at room temperature for 13 h. The reaction was complete (by TLC analysis (10% MeOH in DCM, R f 0.8) and LCMS confirmed), the reaction mixture was concentrated under vacuum, diluted with EtOAc (10 mL), washed with water (2×20 mL), brine (20 mL), dried over anhydrous Na 2 SO 4 , filtered and concentrated. The resulting residue was purified by chromatography (Biotage R snap cartridge, KP-Sil, 25 g, 230-400 silica gel) with 3-4% MeOH in DCM and lyophilized to afford N-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)acetamide 82 (120 mg, 0.267 mmol, 53.5% yield) as an off-white solid.

[0955] 1H-NMR (400MHz, DMSO-d6): δ11.00(s,1H),7.78(t,J=5.60Hz,1H),7.38(t,J=7.60Hz,1H),7.19(d ,J=7.20Hz,1H),7.07(d,J=8.00Hz,1H),5.10(t,J=5.20Hz,1H),4.40(d,J=16.80Hz,1H),4.29(d, J=16.80Hz,1H),3.17(t,J=8.40Hz,4H),2.95(q,J=1.60Hz,3H),2.62-2.53(m,1H),2.48(s,1H),2 .02(d,J=5.20Hz,1H),1.76(s,3H),1.47-1.38(m,6H),1.28-1.22(m,4H),0.84(t,J=6.80Hz,3H).

[0956] LCMS: 443.3 (M+H). Method: Mobile Phase A: 0.1% TFA in H₂O. Mobile Phase B: 0.1% TFA in ACN. Flow Rate: 1.5 mL / min. Column: XBridge C8 (50 × 4.6 mm), 3.5 μm. Rt (min): 1.414; Area %: 98.439.

[0957] HPLC: Method: Mobile phase A: 0.1% TFA in H2O. Mobile phase B: acetonitrile. Flow rate: 2.0 mL / min. Column: X-Bridge C8 (50 × 4.6 mm, 3.5 μm). Rt (min): 2.445; Area %: 98.636.

[0958] Compound 87 in the form of hydrochloride salt

[0959] 3-(4-(((1S,4S)-4-aminocyclohexyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0960]

[0961] Step 1: tert-Butyl ((1S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)carbamate

[0962]

[0963] 3-(4-Bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione 1 (0.5 g, 1.547 mmol, 1 eq), tert-butyl ((1S,4S)-4-aminocyclohexyl)carbamate 2 (0.995 g, 4.64 mmol, 3 eq) and NaOt-Bu (0.446 g, 4.64 mmol, 3 eq) were dissolved in DMF (2 mL) and degassed with nitrogen for 20 min. Pd-PEPPSI-IHept(Cl) (0.075 g, 0.077 mmol, 0.05 eq) was added under nitrogen and the reaction was heated at 110° C. for 4 h. The reaction was complete (by TLC analysis (100% EtOAc R f 0.4) and LCMS confirmation), the reaction mixture was filtered through a bed of celite, washed with THF (10 mL), concentrated to a minimum volume (3 mL), and purified by reverse phase column chromatography ( The mixture was purified by HPLC-MS / ...

[0964] LCMS: 401.1 (M-Boc). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm. Mobile phase: A: 0.1% FA in H2O. B: ACN. Flow rate: 1.5 mL / min. Rt (min): 2.110; Area % -94.51.

[0965] 1 H-NMR (400MHz, DMSO-d6): δ11.02(s,1H),7.28(t,J=7.60Hz,1H),6.94(d,J =6.80Hz,1H),6.79(d,J=8.00Hz,1H),6.61(m,1H),5.14(q,J=5.20Hz,1H), 5.07(d,J=6.40Hz,1H),4.22(q,J=17.20Hz,2H),3.47-3.42(m,2H),2.98-2 .90(m,1H),2.06-2.04(m,1H),1.66(m,6H),1.58-1.54(m,4H),1.39(s,9H).

[0966] Step 2: tert-Butyl ((1S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)cyclohexyl)carbamate

[0967]

[0968] To a solution of tert-butyl ((1S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)carbamate 3 (110 mg, 0.241 mmol, 1 eq), valeraldehyde 4 (0.128 mL, 1.205 mmol, 5 eq) in DCM (4 mL) and DMF (4 mL) at 0°C was added TFA (0.017 ml, 0.217 mmol, 0.9 eq) followed by sodium triacetoxyborohydride (77.0 mg, 0.361 mmol, 1.5 eq). The reaction mixture was warmed to RT and stirred at the same temperature for 16 h. The reaction was complete (by TLC analysis (100% EtOAc R f ~0.5) and LCMS confirmed), the reaction mixture was diluted with ice-cold water (15 mL), extracted with ethyl acetate (2×15 mL), washed with brine (25 mL), dried over anhydrous Na2SO4, filtered and concentrated. The resulting residue was purified by reverse phase column chromatography ( The purified product was purified by HPLC-MS / MS (column: C18 40 μm, 40 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN aqueous mobile phase). Pure fractions from the column were lyophilized to afford tert-butyl ((1S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)cyclohexyl)carbamate 5 (74 mg, 0.140 mmol, 58% yield) as a white solid.

[0969] LCMS: 527.3 (M+H). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm Mobile phase: A: 0.1% FA in H2O B: ACN, Flow rate: 1.5 mL / min. Rt (min): 2.644; Area % -99.9.

[0970] 1H-NMR (400MHz, DMSO-d6): δ10.96(s,1H),7.41(t,J=7.60Hz,1H),7.26(q,J=7.60Hz,2 H),6.97(d,J=6.40Hz,1H),5.08(q,J=5.20Hz,1H),4.31(q,J=17.20Hz,2H),3.55(s,1H ),3.32-2.91(m,3H),2.96-2.80(m,1H),2.60-2.55(m,1H),2.04-2.00(m,1H),1.82-1. 79(m,2H),1.65-1.63(m,2H),1.46-1.40(m,12H),1.27(m,6H),0.81(t,J=7.20Hz,3H).

[0971] Step 3: 3-(4-(((1S,4S)-4-aminocyclohexyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0972]

[0973] To an ice-cold solution of tert-butyl ((1S, 4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)cyclohexyl)carbamate 5 (74 mg, 0.141 mmol, 1 eq) in DCM (4 mL) was added HCl (4 M in EtOAc, 4 mL) and stirred at room temperature for 1.5 h. The reaction progress was monitored by LCMS. The reaction mixture was concentrated, washed with MTBE (2 × 5 mL) and further lyophilized to afford 3-(4-(((1S, 4S)-4-aminocyclohexyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride 87 (44 mg, 0.093 mmol, 97% yield) as an off-white solid.

[0974] LCMS: 427.2 (M-HCl). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm. Mobile phase: A: 0.1% FA in H2O. B: ACN. Flow rate: 1.5 mL / min. Rt (min): 1.634. Area % -85.00.

[0975] HPLC: 97.53%, Rt (min): 9.103. Method: Column: X-Bridge C8 (50×4.6) mm, 3.5 μm, Mobile phase: A: 0.1% TFA in water, Mobile phase: B: ACN, Flow rate: 1.0 mL / min.

[0976] 1 H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),10.98(s,3H),7.45-7.34(m,3H),5.11(dd,J=5.20,13.20Hz,1H),4.41-4.32(m,2H),3.24(m,4H),2 .93-2.92(m,1H),2.63-2.50(m,2H),2.02(t,J=6.80Hz,1H),1.91-1.6 8(m,4H),1.65-1.58(m,4H),1.58-1.21(m,6H),0.80(t,J=6.80Hz,3H).

[0977] Compound 88 in the form of hydrochloride salt

[0978] 3-(4-(((1R,4R)-4-aminocyclohexyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0979]

[0980] Step 1: tert-Butyl ((1R,4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)carbamate

[0981]

[0982] 3-(4-Bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione 1 (0.5 g, 1.547 mmol, 1 eq), tert-butyl ((1R,4R)-4-aminocyclohexyl)carbamate 2 (0.995 g, 4.64 mmol, 3 eq) and NaOt-Bu (0.446 g, 4.64 mmol, 3 eq) were dissolved in DMF (2 mL) and degassed with nitrogen for 20 minutes. Pd-PEPPSI-IHept(Cl) (0.075 g, 0.077 mmol, 0.05 eq) was added under a nitrogen atmosphere and the reaction was heated at 110° C. for 4 h. The reaction was complete (by TLC analysis (100% EtOAc R f 0.4) and LCMS confirmation), the reaction mixture was filtered through celite, washed with THF (10 mL), concentrated to a minimum volume (3 mL), and purified by reverse phase column chromatography ( The mixture was purified by HPLC-MS / ...

[0983] LCMS: 401.1 (M-Boc). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm. Mobile phase: A: 0.1% FA in H2O. B: ACN. Flow rate: 1.5 mL / min. Rt (min): 2.188; Area % -98.18.

[0984] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),7.27(t,J=7.20Hz,1H),6.91(d,J= 7.20Hz,1H),6.80(t,J=8.00Hz,2H),5.29(d,J=6.80Hz,1H),5.12(q,J=4.80H z,1H),4.16(q,J=17.20Hz,2H),2.93-2.89(m,1H),2.70-2.60(m,2H),2.30- 2.20(m,2H),2.09-1.99(m,3H),1.80(m,2H),1.39(s,9H),1.32-1.24(m,4H).

[0985] Step 2: tert-Butyl ((1R,4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)cyclohexyl)carbamate

[0986]

[0987] To a stirred solution of tert-butyl ((1R,4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)carbamate 3 (0.040 g, 0.086 mmol, 1 eq), valeraldehyde (0.076 g, 0.876 mmol, 10 eq) in DCM (1.5 mL) and DMF (1.5 mL) at 0°C was added TFA (0.012 mL, 0.158 mmol, 1.8 eq) and sodium triacetoxyborohydride (56 mg, 0.262 mmol, 3 eq) and the reaction mixture was stirred at room temperature for 16 h. The reaction was complete (by TLC analysis (100% EtOAc R f~0.5) and LCMS confirmed), the reaction mixture was diluted with ice-cold water (15 mL), extracted with ethyl acetate (2×15 mL), washed with brine (25 mL), dried over anhydrous Na2SO4, filtered and concentrated. The resulting residue was purified by reverse phase column chromatography ( The purified product was purified by HPLC-MS / MS (column: C18 40 μm, 40 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN aqueous solution mobile phase). Pure fractions from the column were lyophilized to afford tert-butyl ((1R,4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)cyclohexyl)carbamate 5 (20 mg, 0.038 mmol, 43% yield) as a white solid.

[0988] LCMS: 527.3 (M+H). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm Mobile phase: A: 0.1% FA in H2O B: ACN, Flow rate: 1.5 mL / min. Rt (min): 1.876; Area % -99.04.

[0989] Step 3: 3-(4-(((1R,4R)-4-aminocyclohexyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0990]

[0991] To an ice-cold solution of tert-butyl ((1R, 4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)cyclohexyl)carbamate 5 (20 mg, 0.038 mmol, 1 eq) in DCM (1 mL) was added HCl (4 M in EtOAc, 0.5 mL) and stirred at room temperature for 3 h. The reaction progress was monitored by LCMS. The reaction mixture was concentrated, washed with MTBE (2×5 mL) and lyophilized to afford 3-(4-(((1R, 4R)-4-aminocyclohexyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride 88 (17 mg, 0.036 mmol, 99% yield) as an off-white solid.

[0992] LCMS: 427.2 (M-HCl). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm. Mobile phase: A: 0.1% FA in H2O. B: ACN. Flow rate: 1.5 mL / min. Rt (min): 1.579; Area % -98.76.

[0993] HPLC: 99.10%, Rt (min): 2.381. Method: Column: X-Bridge C8 (50×4.6) mm, 3.5 μm, Mobile phase: A: 0.1% TFA in water, Mobile phase: B: ACN, Flow rate: 2.0 mL / min.

[0994] 1 H-NMR(400MHz,DMSO-d6): δ10.98(s,1H),7.91(br s,3H),7.44-7.32(m,3H),5.13(dd,J=5.20,13.20Hz,1H),4.35-4.07(m,2H),3.11-2.88(m,5H),2.68-2.61(m,2 H),2.02-1.95(m,3H),1.78(s,2H),1.60-1.57(m,2H),1.40-1.31(m,2H),1.24-1.20(m,6H),0.81-0.80(m,3H).

[0995] Compound 89 in the form of hydrochloride salt

[0996] 3-(4-(((1S,4S)-4-(Aminomethyl)cyclohexyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[0997]

[0998] Step 1: tert-Butyl (((1S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)methyl)carbamate

[0999]

[1000] 3-(4-Bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione 1 (1.0 g, 3.09 mmol, 1 eq), tert-butyl (((1S,4S)-4-aminocyclohexyl)methyl)carbamate 2 (1.060 g, 4.64 mmol, 1.5 eq) and NaOt-Bu (0.892 g, 9.28 mmol, 3 eq) were dissolved in DMF (5 mL) and degassed with nitrogen for 20 min. Pd-PEPPSI-IHept(Cl) (0.151 g, 0.155 mmol, 0.05 eq) was added under nitrogen and the reaction was heated at 110° C. for 4 h. The reaction was complete (by TLC analysis (100% EtOAc R f0.4) and LCMS confirmation), the reaction mixture was filtered through a bed of celite, washed with THF (10 mL), concentrated to a minimum volume (3 mL), and purified by reverse phase column chromatography ( The mixture was purified by HPLC-MS / ...

[1001] LCMS: 415.1 (M-Boc). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm. Mobile phase: A: 0.1% FA in H2O. B: ACN. Flow rate: 1.5 mL / min. Rt (min): 2.325. Area % -89.09.

[1002] Step 2: tert-Butyl (((1S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)cyclohexyl)methyl)carbamate

[1003]

[1004] To a solution of tert-butyl (((1S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)methyl)carbamate 3 (80 mg, 0.170 mmol, 1 eq), valeraldehyde 4 (73.2 mg, 0.850 mmol, 5 eq) in DCM (2 mL) and DMF (2 mL) at 0°C was added TFA (0.012 mL, 0.153 mmol, 0.9 eq) followed by sodium triacetoxyborohydride (54 mg, 0.255 mmol, 1.5 eq). The reaction mixture was warmed to RT and stirred at the same temperature for 16 h. The reaction was complete (by TLC analysis (100% EtOAc R f ~0.5) and LCMS confirmed), the reaction mixture was diluted with ice-cold water (15 mL), extracted with ethyl acetate (2×15 mL), washed with brine (25 mL), dried over anhydrous Na2SO4, filtered and concentrated. The resulting residue was purified by reverse phase column chromatography ( (column: C18 40 μm, 40 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN aqueous mobile phase). Pure fractions from the column were lyophilized to afford tert-butyl (((1S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)cyclohexyl)methyl)carbamate 5 (35 mg, 0.060 mmol, 35.5% yield) as an off-white solid.

[1005] LCMS: 541.3 (M+H). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm Mobile phase: A: 0.1% FA in H2O B: ACN, Flow rate: 1.5 mL / min. Rt (min): 2.648; Area % -93.13.

[1006] Step 3: 3-(4-(((1S,4S)-4-(aminomethyl)cyclohexyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[1007]

[1008] To an ice-cold solution of tert-butyl (((1S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)cyclohexyl)methyl)carbamate 5 (35 mg, 0.065 mmol, 1 eq) in DCM (1 mL) was added HCl (4 M in EtOAc, 3 mL) and stirred at room temperature for 1.5 h. The reaction progress was monitored by LCMS. The reaction mixture was concentrated, washed with MTBE (2×5 mL) and lyophilized to afford 3-(4-(((1S,4S)-4-(aminomethyl)cyclohexyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride 89 (22.5 mg, 0.045 mmol, 69% yield) as an off-white solid.

[1009] LCMS: 441.3 (M-HCl). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm. Mobile phase: A: 0.1% TFA in H2O. B: 0.1% TFA in ACN. Flow rate: 1.5 mL / min. Rt (min): 1.408; Area % -94.03.

[1010] HPLC: 94.36%, Rt (min): 8.083. Method: Column: X-Bridge C8 (50×4.6) mm, 3.5 μm, Mobile phase: A: 0.1% TFA in water, Mobile phase: B: ACN, Flow rate: 1.0 mL / min.

[1011] 1 H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.84(s,3H),7.34(d,J=6.40Hz,3H),5.11(dd,J=4.80,12.20Hz,1H),4.38-4.24(m,2H),3.19-3.10(m, 3H),2.92-2.84(m,3H),2.69-2.68(m,2H),1.95(s,1H),1.75(s,1H),1.6 0-1.50(m,4H),1.49-1.30(m,4H),1.24-1.20(m,6H),0.80-0.78(m,3H).

[1012] Compound 90 in the form of a hydrochloride salt

[1013] 3-(4-(((1R,4R)-4-(aminomethyl)cyclohexyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[1014]

[1015] Step 1: tert-Butyl (((1R,4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)methyl)carbamate

[1016]

[1017] To a solution of 3-(4-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione 1 (0.500 g, 1.547 mmol, 1 eq) in DMF (2 mL) was added tert-butyl (((1R,4R)-4-aminocyclohexyl)methyl)carbamate 2 (1.060 g, 4.64 mmol, 3 eq) and sodium tert-butoxide (0.446 g, 4.64 mmol, 3 eq) at room temperature and degassed with nitrogen for 15 minutes. Under nitrogen conditions, Pd-PEPPSI-IHept(Cl) (0.075 g, 0.077 mmol, 0.05 eq) was added to the reaction mixture, degassed for 5 minutes and stirred at 110° C. for 4 hours. After the reaction was completed (confirmed by UPLC analysis), the reaction mixture was filtered through a celite bed and washed with CH 3 CN (5 mL), and the filtrate was concentrated under reduced pressure to obtain a brown thick liquid. The crude compound was purified by reverse phase chromatography ( Column: C18 40 μm, 120 g, eluting the product with 44% ACN in 0.1% HCOOH). Pure fractions were lyophilized to afford tert-butyl (((1R,4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)methyl)carbamate 3 as a yellow solid (0.100 g, 0.118 mmol, 7.60% yield).

[1018] LCMS: 415.2 (M-56). Method: Column: Xbridge C8 (50×4.6 mm) 3.5 μm, Mobile phase: A: 0.1% TFA in H2O, Mobile phase: B: 0.1% TFA in ACN, Flow rate: 1.5 mL / min, Area % -55.37%.

[1019] Step 2: tert-Butyl (((1R,4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)cyclohexyl)methyl)carbamate

[1020]

[1021] To a mixture of tert-butyl (((1R, 4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)methyl)carbamate 3 (0.100 g, 0.213 mmol, 1.0 eq) and valeraldehyde 4 (0.183 g, 2.125 mmol, 1 eq) in DCE (1.5 mL) and DMF (1.5 mL) was added TFA (0.033 mL, 0.425 mmol, 2.0 eq) and stirred at 0 ° C for 10 minutes. To this mixture was added sodium triacetoxyborohydride (0.135 g, 0.638 mmol, 3.0 eq) in portions at 0 ° C, and the resulting colorless turbid material was stirred at room temperature for 16 h (monitored by UPLC). The reaction mixture was quenched with ice-cold water (5 mL) and extracted with dichloromethane (3×10 mL). The combined organic layers were dried over Na2SO4 and concentrated under vacuum to afford the crude material as a light yellow liquid. The crude compound was purified by reverse phase chromatography (eluting with 50% ACN in 0.1% HCOOH) to afford tert-butyl (((1R,4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)cyclohexyl)methyl)carbamate (0.020 g, 0.037 mmol, 17.18% yield) as a white solid.

[1022] 1 H-NMR (400MHz, DMSO-d6): δ11.06(s,1H),7.81(t,J=37.20Hz,1H),7.24(d,J=6.00Hz,2H),5.26(d,J=Hz,1H),4.72(t,J=6.00Hz,2H) ,3.35(d,J=19.60Hz,3H),2.75(d,J=6.40Hz,3H),2.33(d,J=1.60Hz,2H),1.71(d,J=12.00Hz,5H),1.42-1.37(m,14H),0.79(s,3H).

[1023] LCMS: 541.4 (M+H) Method: Column: XBridge C8 (50×4.6 mm) 3.5 μm Mobile phase: A: 0.1% TFA in H 2 O Mobile phase: B: 0.1% TFA in ACN Flow rate: 1.5 mL / min Area % -94.78%.

[1024] HPLC: Method: Column: X-Bridge C8 (50×4.6) mm, 3.5 μm Mobile phase: A: 0.1% TFA in water Mobile phase: B: ACN, Flow rate: 2.0 mL / min, Area %-98.72%.

[1025] Step 3: 3-(4-(((1R,4R)-4-(aminomethyl)cyclohexyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[1026]

[1027] To a stirred solution of tert-butyl (((1R,4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)cyclohexyl)methyl)carbamate 5 (20 mg, 0.037 mmol, 1.0 equiv) in dichloromethane (1 mL) at 0°C was added HCl (4 M in dioxane, 0.5 mL) and then stirred at room temperature for 2 h. The results were analyzed by TLC (10% MeOH; DCM, R f The reaction mixture was concentrated under vacuum at a temperature of less than 40 ° C to obtain an off-white solid. The solid obtained was dissolved in water (5 mL) and washed with MTBE (2 × 5 mL), and the aqueous layer was freeze-dried to obtain 3- (4- (((1R, 4R) -4- (aminomethyl) cyclohexyl) (pentyl) amino) -1- oxoisoindolin-2-yl) piperidine -2,6- dione hydrochloride (17 mg, 0.035 mmol, 96% yield) as an off-white solid.

[1028] 1 H-NMR(400MHz,DMSO-d6): δ10.99(s,1H),7.86(br s,3H),7.43-7.29(m,3H),5.12(d,J=8.80Hz,1H),4.33-4.27(m,2H),3.50-3.30(m,2H),3.12-3.08(m,2H),2.96-2.87(m,1H),2. 68-2.59(m,3H),2.04-2.02(m,1H),1.80-1.75(m,4H),1.50-1.48(m,3H),1.21(s,6H),1.00-0.97(m,2H),0.79(t,J=6.40Hz,3H).

[1029] LCMS: 441.3 (M+H). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm, Mobile phase: A: 0.1% FA in H 2 O, Mobile phase: B: ACN, Flow rate: 1.5 mL / min, Rt-1.526 min, Area: 97.05%.

[1030] HPLC: Method: Column: X-Bridge C8 (50×4.6) mm, 3.5 μm, Mobile phase: A: 0.1% TFA in water, Mobile phase: B: ACN, Flow rate: 2.0 mL / min, Rt-2.287 min, Area: 99.27%.

[1031] Compound 91 in the form of hydrochloride salt

[1032]

[1033] Step 1: tert-Butyl ((1S,4S)-4-formylcyclohexyl)carbamate

[1034]

[1035] To a cooled solution of tert-butyl ((1S,4S)-4-(hydroxymethyl)cyclohexyl)carbamate (200 mg, 0.872 mmol, 1 eq) in DCM (5 mL) was added DMSO (1.5 mL), DIPEA (438 mg, 3.39 mmol, 3.89 eq) and pyridine sulfur trioxide (540 mg, 3.39 mmol, 3.89 eq) in DMSO (1.5 mL) dropwise and stirred at 0° C. for 1 h. The reaction was complete (by TLC (40% ethyl acetate in petroleum ether, R f To the 4-nitro-2-thiazolinone (50mg, 0.870mmol, 1.09mmol) of t-butyl carbamate 2A (200mg, 0.870mmol, 1.01% yield) of crude product ((1S, 4S)-4-formylcyclohexyl)carbamate 2A) (200mg, 0.880mmol, 101% yield) of colorless liquid is added.

[1036] Step 2: tert-Butyl ((1S,4S)-4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamate

[1037]

[1038] To a solution of 3-(4-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione 1 (200 mg, 0.771 mmol, 1 eq) and tert-butyl ((1S, 4S)-4-formylcyclohexyl)carbamate 2A (210 mg, 0.926 mmol, 5 eq) in a mixture of DCE (2 mL) and DMF (0.5 mL) was added TFA (79 mg, 0.694 mmol, 0.9 eq) and Na(OAc) 3BH (245 mg, 1.157 mmol, 1.5 eq) at 0 ° C., and allowed to stir at room temperature for 12 h. After completion of the reaction (confirmed by UPLC), the reaction mixture was quenched with ice water (25 mL) and extracted with DCM (2 × 10 mL). The combined organic layers were concentrated under vacuum to provide a crude product, which was purified by reverse phase chromatography (HPLC). Column: general C18, 40 g snap, 0.1% formic acid in ACN, 20 mL / min, 70-75% ACN in H2O) purification to afford tert-butyl ((1S,4S)-4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamate 3 (105 mg, 0.223 mmol, 28.9% yield) as an off-white solid.

[1039] 1 H-NMR (400MHz, DMSO-d6): δ11.03 (s, 1H), 7.28 (t, J = 8.00Hz, 1H), 6.91 (d, J = 7.20Hz, 1H),6.73(d,J=8.00Hz,2H),5.64(t,J=5.20Hz,1H),5.12(q,J=5.20Hz,1H),4.23(d,J =17.20Hz,1H),4.12(d,J=17.20Hz,1H),3.49(s,1H),3.03(t,J=6.00Hz,2H),2.92(t, J=8.00Hz,1H),2.06-2.02(m,1H),1.69(s,1H),1.50(t,J=13.20Hz,8H),1.39(s,9H).

[1040] LCMS: 470.0 (M+H), Rt (min): 2.306, Area %: 99.775.

[1041] HPLC: Rt (min): 4.229, Area %: 99.789.

[1042] Step 3: tert-Butyl ((1S,4S)-4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)methyl)cyclohexyl)carbamate

[1043]

[1044] To a solution of tert-butyl ((1S,4S)-4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamate (90 mg, 0.191 mmol, 1 eq) in DCE (5 mL) and DMF (0.5 mL) was added valeraldehyde (82 mg, 0.956 mmol, 5 eq), TFA (19.63 mg, 0.172 mmol, 0.9 eq) and Na(OAc)BH (48.6 mg, 0.230 mmol, 1.5 eq) at 0° C. and allowed to stir at room temperature for 12 h. After completion of the reaction (confirmed by LCMS), the reaction mixture was quenched with ice water (25 mL) and extracted with DCM (2×10 mL). The combined organic layers were concentrated under vacuum to afford the crude product, which was purified by Isolera (column size: Biotage R snap cartridge, KP-Sil, 10 g, silica gel - 230-400 mesh) using 70-80% ethyl acetate in petroleum ether. Pure fractions were collected separately and concentrated under vacuum to afford tert-butyl ((1S,4S)-4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)methyl)cyclohexyl)carbamate (30 mg, 0.055 mmol, 29.0% yield) as a white solid.

[1045] 1H-NMR (400MHz, DMSO-d6): δ10.99(s,1H),7.37(t,J=7.60Hz,1H),7.18(d,J=7.20Hz,1H),7.10(d,J=8.00H z,1H),6.66(d,J=7.20Hz,1H),5.11(q,J=4.80Hz,1H),4.38(d,J=16.80Hz,1H),4.28(d,J=17.20Hz,1H),3. 45(s,1H),3.34-3.16(m,2H),3.11(d,J=7.20Hz,2H),2.90(d,J=12.00Hz,1H),2.59(d,J=19.60Hz,2H),2.0 2(t,J=5.20Hz,1H),1.46(t,J=7.20Hz,4H),1.38(s,15H),1.24(t,J=10.00Hz,5H),0.84(t,J=6.80Hz,3H).

[1046] LCMS: 485.9 (M-56), Rt (min): 2.828, Area %: 99.796.

[1047] HPLC: Rt (min): 4.910, Area %: 98.887.

[1048] Step 4: 3-(4-((((1S,4S)-4-aminocyclohexyl)methyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl

[1049]

[1050] To an ice-cold solution of tert-butyl ((1S,4S)-4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)methyl)cyclohexyl)carbamate (30 mg, 0.055 mmol) in DCM (2 mL) was added HCl (4 M in ethyl acetate) (0.5 mL, 16.46 mmol) at 0° C. and stirred at room temperature for 6 h. After completion of the reaction (confirmed by LCMS), the reaction mixture was concentrated and washed with MTBE (10 mL) to afford the crude product, which was lyophilized to afford 3-(4-((((1S,4S)-4-aminocyclohexyl)methyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl (30 mg, 0.063 mmol, 113% yield) as an off-white solid.

[1051] 1H-NMR (400MHz, DMSO-d6): δ11.00(s,1H),7.78(s,6H),7.39(t,J=8.00Hz,1H) ,7.22(d,J=7.60Hz,1H),7.22(d,J=7.60Hz,1H),5.12(q,J=5.20Hz,1H),4.33 (dd,J=16.80,42.40Hz,2H),3.15(q,J=7.60Hz,5H),2.05-2.01(m,1H),1.59( s,5H),1.45(d,J=5.60Hz,6H),1.25(d,J=4.80Hz,4H),0.84(t,J=7.20Hz,3H).

[1052] LCMS: 442.0 (M+H), method: mobile phase: A: 0.1% formic acid in H2O solution B: ACN, column: Atlantis dC18 (50×4.6) 5μm, flow rate: 1.5mL / min; Rt (min): 1.691, area%: 99.845.

[1053] HPLC: Method: Mobile phase: A: 0.1% FA in H2O, Mobile phase: B: ACN, Column: Atlantis dC18 (50×4.6 mm) 5 μm, Flow rate: 1.5 mL / min, Rt (min): 2.839, Area %: 98.026.

[1054] Compound 92 in the form of a hydrochloride salt

[1055]

[1056] Step 1: tert-Butyl ((1R,4R)-4-formylcyclohexyl)carbamate

[1057]

[1058] To a cooled solution of tert-butyl ((1R,4R)-4-(hydroxymethyl)cyclohexyl)carbamate (500 mg, 2.180 mmol, 1 eq) in DCM (11 mL) was added DMSO (3.7 mL), DIPEA (0.388 mL, 2.180 mmol, 3.89 eq) and pyridine sulfur trioxide (347 mg, 2.180 mmol, 3.89 eq) in DMSO (3.7 mL) dropwise and stirred at 0°C for 1 h. The reaction was complete (by TLC (40% ethyl acetate in petroleum ether, R fTo the 4-nitro-2-thiazolinyl group, 1.5N HCl (0.4) was added 1.5N HCl (20 mL) to quench the reaction mixture and extract with DCM (2 × 20 mL). The organic layer was dried over anhydrous Na2SO4 and concentrated under vacuum to provide the crude product ((1R, 4R)-4-formylcyclohexyl) tert-butyl carbamate 2A (480 mg, 2.112 mmol, 97% yield) as a yellow liquid.

[1059] 1 H-NMR (400MHz, DMSO-d6): δ9.55(s,1H),3.64-3.50(m,1H),3.16-3.11(m,2H),2.16-2.11( m,1H),1.99(d,J=57.60Hz,2H),1.88(t,J=24.40Hz,3H),1.38(s,10H),1.18-1.26(m,11H).

[1060] LCMS: 172.4 (M-56), Rt (min): 2.593, Area %: 59.513.

[1061] Step 2: tert-Butyl ((1R,4R)-4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamate

[1062]

[1063] At 0 ° C, to 3- (4-amino -1- oxoisoindolin-2-yl) piperidine -2,6- dione 1 (350 mg, 1.350 mmol, 1 equivalent) and ((1R, 4R) -4- formylcyclohexyl) carbamic acid tert-butyl ester 2A (368 mg, 1.620 mmol, 5 equivalents) in a mixture of DCE (1.5 mL) and DMF (0.5 mL) was added TFA (139 mg, 1.215 mmol, 0.9 equivalents) and Na (OAc) 3BH (429 mg, 2.025 mmol, 1.5 equivalents), and it was stirred at room temperature for 12 h. After the reaction was completed (confirmed by TLC), the reaction mixture was quenched with ice water (25 mL) and extracted with DCM (2 × 10 mL). The combined organic layer was concentrated under vacuum to provide a crude product, which was purified by reverse phase chromatography (HPLC). 8-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamic acid tert-butyl ester 3 (410 mg, 0.834 mmol, 61.8% yield) was obtained by lyophilization of the purified product using a column: general C18, 40 g snap, 0.1% formic acid in ACN, 20 mL / min, 70-75% ACN in HO) and pure fractions were lyophilized to afford tert-butyl ((1R,4R)-4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamate 3 as a white solid (410 mg, 0.834 mmol, 61.8% yield).

[1064] 1 H-NMR (400MHz, DMSO-d6): δ11.00(s,1H),7.27(t,J=7.60Hz,1H),6.91(d,J=7.60Hz,1H),6.71(q,J=8.00Hz ,2H),5.62(t,J=5.60Hz,1H),5.12(q,J=4.80Hz,1H),4.23(d,J=17.20Hz,1H),4.12(d,J=17.20Hz,1H),3.1 9(d,J=8.00Hz,1H),2.98-2.89(m,3H),2.60(t,J=22.00Hz,2H),2.35-2.28(m,2H),2.06-2.02(m,1H),1.81 (t,J=20.00Hz,4H),1.50(t,J=7.20Hz,2H),1.37(s,9H),1.12(q,J=11.60Hz,3H),0.99(t,J=12.00Hz,2H),

[1065] LCMS: 415.3 (M-56), Rt (min): 2.127, Area %: 95.713.

[1066] HPLC: Rt (min): 4.211, Area %: 98.834.

[1067] Step 3: tert-Butyl ((1R,4R)-4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)methyl)cyclohexyl)carbamate

[1068]

[1069] To a solution of tert-butyl ((1R,4R)-4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamate (250 mg, 0.531 mmol, 1 eq) in DCE (2.5 mL) and DMF (0.5 mL) was added valeraldehyde (229 mg, 2.66 mmol, 5 eq), TFA (54.5 mg, 0.478 mmol, 0.9 eq) and Na(OAc)BH (169 mg, 0.797 mmol, 1.5 eq) at 0° C. and allowed to stir at room temperature for 12 h. After completion of the reaction (confirmed by LCMS), the reaction mixture was quenched with ice water (25 mL) and extracted with DCM (2×10 mL). The combined organic layers were concentrated under vacuum to afford the crude product, which was purified by Isolera (column size: Biotage R snap cartridge, KP-Sil, 25 g, silica gel - 230-400 mesh) using 70-80% ethyl acetate in petroleum ether. Pure fractions were collected separately and concentrated under vacuum to afford tert-butyl ((1R,4R)-4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)methyl)cyclohexyl)carbamate 5 (124 mg, 0.229 mmol, 43.2% yield) as a yellow solid.

[1070] 1 H-NMR (400MHz, DMSO-d6): δ10.97(s,1H),7.37(t,J=8.00Hz,1H),7.13(dd,J=8.00, 35.00Hz,2H),6.63(d,J=8.00Hz,1H),5.09(t,J=5.20Hz,1H),4.31(t,J=16.80Hz,2 H),3.31-3.15(m,3H),3.05-3.03(m,2H),2.00(s,1H),1.72(d,J=9.60Hz,5H),1.43 -1.36(m,14H),1.26-1.20(m,5H),0.97(d,J=27.20Hz,4H),0.83(t,J=6.80Hz,3H).

[1071] LCMS: 541.3 (M+H), Rt (min): 2.849, Area %: 95.954.

[1072] HPLC: Rt (min): 4.907, Area %: 95.462.

[1073] Step 4: 3-(4-((((1R,4R)-4-aminocyclohexyl)methyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl

[1074]

[1075] To an ice-cold solution of tert-butyl ((1R,4R)-4-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)methyl)cyclohexyl)carbamate (120 mg, 0.222 mmol) in DCM (5 mL) was added HCl (4 M in ethyl acetate) (0.12, 3.95 mmol) at 0° C. and stirred at room temperature for 6 h. After completion of the reaction (confirmed by LCMS), the reaction mixture was concentrated, washed with MTBE (10 mL) and dried to afford 3-(4-((((1R,4R)-4-aminocyclohexyl)methyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl 92 (30 mg, 0.061 mmol, 27.3% yield) as a light yellow solid.

[1076] 1 H-NMR (400MHz, DMSO-d6): δ11.00(s,1H),7.83(s,3H),7.39(t,J=7.60Hz,1H),7.20(d,J=7.20Hz,1H),7.11(d,J=8.00H z,1H),5.11(q,J=4.80Hz,1H),4.39(d,J=16.80Hz,1H),4.28(d,J=16.80Hz,1H),3.18(d,J=4.80Hz,2H),3.18(d,J=4.80 Hz,2H),2.93-2.88(m,2H),2.68(t,J=2.00Hz,1H),2.62(d,J=2.80Hz,1H),2.02(d,J=5.60Hz,1H),1.90(d,J=10.40Hz,2 H),1.77(d,J=12.00Hz,2H),1.41(t,J=6.80Hz,3H),1.17-1.20(m,6H),0.97(d,J=12.00Hz,2H),0.83(t,J=6.80Hz,3H).

[1077] LCMS: 441.3 (M+H), method: mobile phase: A: 0.1% FA in H2O, mobile phase: B: ACN, column: Atlantis dC18 (50×4.6 mm) 5 μm, flow rate: 1.5 mL / min, Rt (min): 1.695, area %: 96.479.

[1078] HPLC: Method: A: 0.1% TFA in H2O, B: ACN, Flow rate: 2.0 mL / min

[1079] Column: Xbridge C8 (50×4.6) mm, 3.5 μm, Rt (min): 2.950, Area %: 95.528.

[1080] Compound 93 in the form of a hydrochloride salt

[1081] (E)-3-(4-((4-Aminobutyl)(pent-3-en-1-yl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[1082]

[1083] Step 1: (E)-Pent-3-enal

[1084]

[1085] To a stirred solution of (E)-pent-3-enoic acid methyl ester 2 (2.5 g, 21.90 mmol, 1.0 eq) in CHCl (50 mL) was added a hexane solution of DIBAL-H (17.52 mL, 17.52 mmol, 0.8 eq) dropwise at -78°C. The resulting reaction mixture was then stirred at 78°C for 5 minutes. The reaction was complete (by TLC analysis (20% EtOAc in petroleum ether, R f After 40 ℃ of drying (confirmed), the reaction mixture was quenched with 1.5N HCl solution (10 mL) and extracted with DCM (20 mL). The separated organic layer was washed with brine (20 mL), dried over Na2SO4, filtered and used in the next step without concentration.

[1086] Step 2: tert-Butyl (E)-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pent-3-en-1-yl)amino)butyl)carbamate

[1087]

[1088] To a stirred solution of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 1 (0.5 g, 1.161 mmol, 1.0 eq) and (E)-pent-3-enal 3 (1.954 g, 23.23 mmol, 20.0 eq) in a mixture of CHCl (10 mL) and DMF (5 mL) was added AcOH (0.332 mL, 5.81 mmol, 5.0 eq) under a nitrogen atmosphere and stirred for 1 h. To this mixture was added NaCNBH (0.292 g, 4.65 mmol, 4.0 eq) at 0 ° C. The resulting reaction mixture was stirred at room temperature overnight. The reaction was complete (by TLC and UPLC analysis (100% EtOAc, R f The reaction mixture was diluted with DCM (30 mL), washed with water (2×30 mL), 10% LiCl solution (30 mL), and brine (30 mL), dried over Na 2 SO 4 , filtered, and concentrated to afford the crude product as a light yellow liquid. The crude compound was purified by preparative HPLC (column: X-Bridge-C18 19.1×250, mobile phase: 0.1% FA in water / ACN, flow rate: 15 mL / min) to afford tert-butyl (E)-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pent-3-en-1-yl)amino)butyl)carbamate 4 (35 mg, 0.068 mmol, 5.87% yield) as an off-white solid.

[1089] 1 H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.36-7.40(m,1H),7.18-7.20(m,1H) ),7.06-7.08(m,1H),6.78(s,1H),5.40-5.42(m,3H),5.09-5.13(m,1H),4.26 -4.42(m,2H),3.41-3.51(m,3H),3.19-3.22(m,4H),2.88-2.92(m,3H),2.11- 2.13(m,2H),2.03(m,1H),1.62-1.63(m,3H),1.59-1.60(m,2H),1.37(s,9H),

[1090] LCMS: 499.2 (M+H), Rt (min): 2.492, area-97%.

[1091] Step 3: (E)-3-(4-((4-aminobutyl)(pent-3-en-1-yl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[1092]

[1093] To a stirred solution of tert-butyl (E)-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pent-3-en-1-yl)amino)butyl)carbamate 4 (65 mg, 0.130 mmol, 1.0 equiv) in ethyl acetate (1 mL) at 0°C was added HCl (4 M in EtOAc, 2 mL) and then stirred at room temperature for 3 h. The reaction progress was monitored by TLC (100% EtOAc). The reaction mixture was concentrated under vacuum at below 40°C to afford an off-white semisolid. The solid obtained was dissolved in water (2 mL) and washed with MTBE (2 mL), and additional water (2 mL) was added and lyophilized to afford (E)-3-(4-((4-aminobutyl)(pent-3-en-1-yl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl (30 mg, 0.067 mmol, 79% yield) as an off-white solid.

[1094] 1 H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.84(s,3H),7.09-7.43(m,3H),5.40-5.41(m,2H),5.10-5.14(m,1H),4.32-4.39(m,2H), 3.23-3.48(m,4H),2.90-2.93(m,1H),2.74-2.78(m,2H),2.53-2.68(m,1H),2.12-2.14(m,2H),2.02-2.09(m,2H),1.52-1.60(m,7H)

[1095] LCMS: 399.2 (M+H). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm, Mobile phase: A: 0.1% FA in H2O, Mobile phase: B: acetonitrile, Flow rate: 1.5 mL / min

[1096] HPLC: Method: Mobile phase: A: 0.1% FA in water, Mobile phase: B: ACN, Flow rate: 2.0 mL / min, Column: X-Bridge C8 (50×4.6) mm, 3.5 μm, RT: 2.301 min, Area: 97.652%.

[1097] Trifluoroacetate salt form of compound 94

[1098] (Z)-3-(4-((4-Aminobutyl)(pent-3-en-1-yl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[1099]

[1100] Steps 1 and 2: 3-(1-oxo-4-(pent-3-yn-1-ylamino)isoindolin-2-yl)piperidine-2,6-dione

[1101]

[1102] At 0 ° C, to a solution of pent-3-yn-1-ol 2 (2.0 g, 23.78 mmol, 1.0 equivalent) and CH2Cl2 (100 mL) was added Dess-Martin periodinane (11.09 g, 26.2 mmol, 1.1 equivalents) and water (0.05 mL), and the resulting white suspension was stirred at room temperature for 1 h. After completion of the reaction (confirmed by TLC (20% EtOAc in petroleum ether), the reaction mixture was cooled to -78 ° C and diluted with n-pentane (50 mL) and filtered through a bed of silica gel. The bed was washed with n-pentane (20 mL), and the filtrate was concentrated to an amount of ~ 50% under vacuum at 25 ° C, and the solution of the resulting compound 3 was used directly in the next step.

[1103] In another RB flask, at room temperature, to a stirred solution of 3-(4-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione 1 (4.00 g, 15.43 mmol, 0.649 equivalent) in DMF (40 mL) was added a solution of the above compound 3. Acetic acid (1.753 mL, 30.6 mmol, 1.289 equivalent) was added to the reaction mixture and stirred at room temperature for 2 hours. At 0 ° C, sodium cyanoborohydride (1.520 g, 24.18 mmol, 1.017 equivalent) was added to the reaction mixture and stirred at room temperature overnight. The reaction was complete (by TLC and UPLC analysis (100% EtOAc, R f ~0.7) confirmed), the reaction mixture was diluted with DCM (100 mL) and washed with water (2×50 mL), dried over Na 2 SO 4 , filtered and concentrated to obtain the crude product as a light yellow liquid. The crude compound was purified by reverse phase ( Column: C18 40 μm, 120 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN in water mobile phase, eluting the product with 70% ACN in 0.1% HCO2H) to obtain 3-(1-oxo-4-(pent-3-yn-1-ylamino)isoindolin-2-yl)piperidine-2,6-dione 4 (1.0 g, 3.03 mmol, 12.75% yield) as a light yellow solid.

[1104] 1 H-NMR (400MHz, DMSO-d6): δ11.01(s,1H),7.30(t,J=8.00Hz,1H),6.96(d,J=7.20Hz, 1H),6.78(d,J=6.00Hz,1H),5.72(t,J=6.00Hz,1H),5.09-5.14(m,1H),4.23(d,J=16 .80Hz,1H),4.12(d,J=17.20Hz,1H),3.25-3.33(m,2H),2.93(m,1H),2.50-2.51(m,1 H),2.40-2.42(m,2H),2.27-2.38(m,1H),2.02-2.06(m,1H),1.75(t,J=2.00Hz,3H),

[1105] LCMS: 326.1 (M+H), Rt (min): 1.986, Area-98.611%.

[1106] Step 3: tert-Butyl (tert-butoxycarbonyl)(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pent-3-yn-1-yl)amino)butyl)carbamate

[1107]

[1108] To a stirred mixture of 3-(1-oxo-4-(pent-3-yn-1-ylamino)isoindolin-2-yl)piperidine-2,6-dione 4 (0.5 g, 1.537 mmol, 1.0 equiv) and tert-butyl (tert-butoxycarbonyl)(5-oxopentyl)carbamate 5 (0.88 g, 3.07 mmol, 2.0 equiv) in CHCl (15 mL) and DMF (15 mL) was added trifluoroacetic acid (0.106 mL, 1.383 mmol, 0.9 equiv) and stirred for 10 minutes at 0° C. Sodium triacetoxyborohydride (0.49 g, 2.305 mmol, 1.5 equiv) was added portionwise at 0° C. and the resulting colorless, turbid material was stirred at room temperature for 16 h. After the reaction was completed (confirmed by UPLC analysis), the reaction mixture was quenched with ice-cold water (50 mL) and extracted with dichloromethane (2×50 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 and concentrated under vacuum to obtain the crude product as a brown liquid. The crude compound was purified by ( Column: C18 40 μm, 120 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN in water mobile phase, eluting the product with 70% ACN in 0.1% HCO2H) to afford tert-butyl (tert-butoxycarbonyl)(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pent-3-yn-1-yl)amino)butyl)carbamate 6 (0.4 g, 0.649 mmol, 42.2% yield) as an off-white solid.

[1109] 1 H-NMR (400MHz, DMSO-d6): δ10.99(s,1H),7.39(t,J=10.00Hz,1H),7.23(d,J=9.60H z,1H),7.12(d,J=10.80Hz,1H),5.08-5.14(m,1H),4.41(d,J=22.80Hz,1H),4.31(d, J=22.40Hz,1H),3.40-3.47(m,2H),3.23-3.33(m,2H),2.87-2.93(m,1H),2.28-2.45 (m,1H),2.01-2.08(m,2H),1.69(t,J=2.00Hz,3H),1.49(s,18H),1.27-1.44(m,8H),

[1110] LCMS: 597.4 (M+H), Rt (min): 2.482, Area-96.894%.

[1111] Step 4: tert-Butyl (Z)-(tert-butoxycarbonyl)(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pent-3-en-1-yl)amino)butyl)carbamate

[1112]

[1113] To a stirred solution of tert-butyl (tert-butoxycarbonyl)(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pent-3-yn-1-yl)amino)butyl)carbamate 6 (150 mg, 0.251 mmol, 1.0 equiv), quinoline (39.0 mg, 0.302 mmol, 1.2 equiv) in DMF (2.0 mL) was added Lindela's catalyst (12 mg, 0.113 mmol, 0.449 equiv) and stirred at room temperature for 16 h under a hydrogen atmosphere. After completion of the reaction (confirmed by UPLC analysis), the reaction mixture was filtered through a bed of celite and washed with DMF (2 mL), and the filtrate was purified by reverse phase chromatography ( Column: C18 40 μm, 120 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN in water mobile phase, eluting the product with 70-75% ACN in 0.1% HCOOH) to afford tert-butyl (Z)-(tert-butoxycarbonyl)(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pent-3-en-1-yl)amino)butyl)carbamate 7 (120 mg, 0.182 mmol, 72.2% yield) as a light yellow gummy solid.

[1114] 1 H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.39(t,J=3.20Hz,1H),7.21(d,J=4.80Hz,1H),7.11(d, J=8.00Hz,1H),5.38-5.45(m,2H),5.09-5.14(m,1H),4.39(d,J=16.80Hz,1H),4.28(d,J=17.20Hz ,1H),3.36-3.47(m,3H),3.18-3.24(m,4H),3.18-3.20(m,1H),2.50-2.53(m,2H),2.44-2.50(m,1 H),2.16-2.18(m,2H),2.00-2.01(m,1H),1.59(d,J=4.80Hz,3H),1.49(s,18H),1.27-1.44(m,2H),

[1115] LCMS: 599.3 (M+H). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm, Mobile phase: A: 0.1% FA in H2O, Mobile phase: B: acetonitrile, Flow rate: 1.5 mL / min

[1116] HPLC: Method: Mobile phase: A: 0.1% TFA in water, Mobile phase: B: methanol, Flow rate: 1.0 mL / min, Column: Atlantis dC18 (250×4.6) mm, 5 μm, RT: 16.959 min, Area: 90.666%.

[1117] Step 5: (Z)-3-(4-((4-aminobutyl)(pent-3-en-1-yl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[1118]

[1119] To a stirred solution of tert-butyl (Z)-(tert-butoxycarbonyl)(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pent-3-en-1-yl)amino)butyl)carbamate 7 (60 mg, 0.100 mmol) in CHCl (1 mL) at 0°C was added TFA (2 mL, 26.0 mmol), followed by stirring at room temperature overnight. After completion of the reaction (confirmed by UPLC), the reaction mixture was concentrated under reduced pressure at 40°C to obtain a light yellow viscous liquid. The crude compound was purified by preparative HPLC (column: Atlantis 19x250 mm, mobile phase: 0.1% TFA in water / ACN, flow rate: 2 mg / mL) to afford (Z)-3-(4-((4-aminobutyl)(pent-3-en-1-yl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, TFA 94 (12 mg, 0.022 mmol, 22.15% yield) as a light yellow solid.

[1120] 1H-NMR (400MHz, DMSO-d6): δ11.00(s,1H),7.61(s,3H),7.41(t,J=7.60Hz,1H),7.24(d,J=6.80Hz,1H),7 .14(d,J=7.60Hz,1H),5.44-5.46(m,1H),5.38-5.39(m,1H),5.11-5.15(m,1H),4.39(d,J=16.80Hz,1H) ,4.28(d,J=16.80Hz,1H),3.22(t,J=8.40Hz,4H),2.79-2.80(m,1H),2.68-2.77(m,2H),2.63-2.68(m,2 H),2.50-2.51(m,1H),2.33-2.34(m,2H),2.17-2.19(m,1H),1.50(t,J=2.00Hz,3H),1.48-1.49(m,3H),

[1121] LCMS: 399.2 (M+H). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm, Mobile phase: A: 0.1% FA in H2O, Mobile phase: B: acetonitrile, Flow rate: 1.5 mL / min

[1122] HPLC: Method: Mobile phase: A: 0.1% TFA in water, Mobile phase: B: ACN, Flow rate: 1.0 mL / min, Column: Atlantis dC18 (250×4.6) mm, 5 μm, RT: 7.709 min, Area: 94.84%.

[1123] Compound 95 in the form of a hydrochloride salt

[1124] 3-(4-((4-Aminobutyl)(pent-3-yn-1-yl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl

[1125]

[1126] Steps 1 and 2: tert-Butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pent-3-yn-1-yl)amino)butyl)carbamate

[1127]

[1128] To a solution of pent-3-yn-1-ol 3 (2 g, 23.78 mmol, 1 equivalent) in DCM (40 mL) was added Dess-Martin periodinane (11.09 g, 26.2 mmol, 1.1 equivalents) at 0 ° C., and the resulting white suspension was stirred at room temperature for 1 h. The reaction mixture was quenched with a mixture of 5 g of saturated NaHCO (50 mL) solution and extracted with DCM (2 × 15 mL). To the combined organic layers was added tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate (0.500 g, 1.162 mmol, 1 equivalent) and acetic acid (0.714 g, 11.627 mmol) and stirred for 2 h. Then add NaCNBH (0.463g, 11.627mmol, 10 equivalents), and make it stir at room temperature 24h.After reaction is completed (confirmed by UPLC), reaction mixture is quenched with water (40mL) and extracted with DCM (2 × 40mL).By the organic layer merged through anhydrous Na sO dry and concentrate under vacuum to obtain crude product, by using the Isolera (Biotage R snap cartridge, KP-Sil, 25g, 230-400 silica mesh) purifying of the petroleum ether solution of 60-70% ethyl acetate.Pure fractions are concentrated under vacuum, to obtain (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl) (pent-3-alkynes-1-yl) amino) butyl) tert-butyl carbamate 3 (150mg, 0.113mmol, 0.474% productive rate) as off-white solid.

[1129] LCMS: 497.2 (M+H), Rt (min): 2.384, Area %: 37.322.

[1130] Step 3: 3-(4-((4-Aminobutyl)(pent-3-yn-1-yl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl

[1131]

[1132] To an ice-cold solution of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pent-3-yn-1-yl)amino)butyl)carbamate 3 (150 mg, 0.118 mmol, 1 eq) in DCM (3 mL) at 0 °C was added HCl (4 M in ethyl acetate, 0.294 mL, 1.178 mmol, 10 eq) and stirred at room temperature for 12 h. After completion of the reaction (confirmed by LCMS), the reaction mixture was concentrated and washed with MTBE (10 mL) to afford the crude product, which was purified by preparative HPLC (column: X-Bridge-C18 19.1X250; method: 0.1% HCl in water / ACN; flow rate: 15 mL / min) to afford 3-(4-((4-aminobutyl)(pent-3-yn-1-yl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl 95 (7.5 mg, 0.017 mmol, 14.16% yield) as a white solid.

[1133] 1 H-NMR (400MHz, DMSO-d6): δ11.00(s,1H),7.78(s,1H),7.41(t,J=7.60Hz,1H),7.25(d,J=7.20H z,1H),7.15(d,J=8.00Hz,1H),5.15-5.10(m,1H),4.45-4.30(m,2H),3.33(t,J=7.20Hz,2H),3. 24(t,J=6.40Hz,3H),2.93(s,1H),2.77-2.68(m,2H),2.33-2.28(m,2H),2.04(t,J=5.20Hz,1H) ,1.70(t,J=2.40Hz,2H),1.52(d,J=4.00Hz,4H),1.24(t,J=3.20Hz,1H),0.84(t,J=7.20Hz,1H).

[1134] LCMS: 397.3 (M+H), method: mobile phase: A: 0.1% TFA in H2O, mobile phase: B: 0.1% TFA in ACN, column: X Bridge C8 (50×4.6 mm) 3.5 μm, flow rate: 1.5 mL / min, Rt (min): 1.340, area %: 96.271.

[1135] HPLC: Method: Mobile phase: A: 0.1% TFA aqueous solution, Mobile phase: B: ACN, Column: X-Bridge C8 (50×4.6) mm, 3.5 μm, Flow rate: 2.0 mL / min, Rt (min): 2.314, Area %: 98.115.

[1136] Compound 96 in the form of a hydrochloride salt

[1137] 3-(4-(Bis(4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1138]

[1139] Step 3: Di-tert-butyl (((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)azanediyl)bis(butane-4,1-diyl))dicarbamate

[1140]

[1141] To a solution of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 4 (250 mg, 0.581 mmol), tert-butyl (tert-butoxycarbonyl)(4-oxobutyl)carbamate 5A (334 mg, 1.161 mmol, 2 eq) and TFA (0.179 mL, 2.323 mmol, 4 eq) in a mixture of dichloromethane (0.5 mL, 20 vol) and DMF (0.25 mL, 10 vol) was added sodium triacetoxyborohydride (492 mg, 2.323 mmol, 4 eq) and stirred at room temperature for 16 h. The reaction was complete (by TLC analysis (EtOAc, R f The mixture was stirred for 2 hours at room temperature for 1 hr at 4 0 ℃ for 1 hr at 8 0 ℃ for 2 h. After 3 0 s, the mixture was stirred for 2 hours at 4 0 ℃ for 3 hr at 8 0 ℃ for 4 0 s. After 3 0 s, the mixture was stirred for 2 hours at 8 0 ℃ for 3 0 s. After 4 0 s, the mixture was stirred for 2 hours at 8 0 ℃ for 3 0 s. After 4 0 s, the mixture was stirred for 2 hours at 8 0 ℃ for 3 0 s. The mixture was stirred for 2 hours at 4 0 s. After 3 0 s, the mixture was stirred for 2 hours at 8 0 ℃ for 3 0 s. The mixture was stirred for 2 hours at 8 0 ℃ for 3 0 s. The mixture was stirred for 2 hours at 8 0 ℃ for 3 0 s. After 4 0 s, the mixture was stirred for 2 hours at 8 0 ℃ for 3 0 s. The mixture was stirred for 2 hours at 8 0 ℃ for 3 0 s. The mixture was stirred for 2 hours at 8 0 ℃ for 3 0 s.

[1142] 1H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.37(t,J=7.60Hz,1H),7.20(d,J=7.20Hz,1H),7.08(d,J=8.00Hz,1H),6.78(d,J=5.20Hz,1H),5.11(q ,J=5.20Hz,1H),4.39(d,J=17.20Hz,1H),4.28(d,J=16.80Hz,1H),3.34 (s,2H),3.18(d,J=6.80Hz,4H),2.90(s,4H),2.00(s,2H),1.40(s,33H).

[1143] LC-MS: 702.4 (M+H), Rt (min): 2.83, Area %: 99.289.

[1144] Step 4: 3-(4-(bis(4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1145]

[1146] To an ice-cold solution of di-tert-butyl (((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)azanediyl)bis(butane-4,1-diyl))dicarbamate 5 (150 mg, 0.249 mmol) in DCM (5 mL) was added HCl (4 M in dioxane) (3 mL, 12.00 mmol) and stirred at room temperature for 2 h. After completion of the reaction as confirmed by TLC analysis (100% EtOAc), the reaction mixture was concentrated and lyophilized to afford 3-(4-(bis(4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl 96 (82 mg, 67.8% yield) (2.4 g, 98%) as a light yellow solid.

[1147] 1 H-NMR (400MHz, DMSO-d6): δ11.02(s,1H),8.07(s,6H),7.52(s,3H),5.12(q,J=5.20Hz,1H),4.65(s,8H),3.42-3.43(m 1H), 2.74 (d, J = 5.20Hz, 4H), 2.62 (d, J = 16.80Hz, 1H), 2.07 (t, J = 5.60Hz, 1H), 1.56 (d, J = 4.80Hz, 8H).

[1148] 1H-NMR (400MHz, CD3OD): δ7.93(s,2H),7.80(s,1H),5.24(q,J=4.80Hz,1H),4.48(q,J=400.80Hz,2 H), 3.66 (t, J = 26.40Hz, 4H), 3.00-2.92 (m, 6H), 2.68-2.61 (m, 1H), 2.28-2.24 (m, 1H), 1.69 (s, 1H).

[1149] LCMS: 402.3 (M+H). Method: Mobile Phase A: 0.1% TFA in H2O. Mobile Phase B: 0.1% TFA in ACN. Flow Rate: 1.5 mL / min. Column: XBridge C8 (50 × 4.6 mm), 3.5 μm. Rt (min): 0.887; Area %: 97.781.

[1150] HPLC: Method: Mobile phase A: 0.1% TFA in H2O. Mobile phase B: acetonitrile. Flow rate: 2.0 mL / min. Column: X-Bridge C8 (50 × 4.6 mm, 3.5 μm). Rt (min): 1.08; Area %: 95.532.

[1151] Compound 97 in the form of a hydrochloride salt

[1152] 3-(4-((4-Aminobutyl)(3-aminopropyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1153]

[1154] Step 1: tert-Butyl (3-oxopropyl)carbamate

[1155]

[1156] Under nitrogen atmosphere, at -78 DEG C, over 15 minutes, a solution of oxalyl chloride (0.599mL, 6.85mmol, 1.2 equivalents) in DCM (10mL) was added dropwise to a solution of DMSO (0.972mL, 13.70mmol, 2.4 equivalents) in DCM (5mL), and stirred at the same temperature for 15 minutes. Over 15 minutes, a solution of tert-butyl (3-hydroxypropyl) carbamate 5 (1g, 5.71mmol, 1 equivalent) in DCM (5mL) was added dropwise thereto. After stirring the reaction mixture for 30 minutes at -78 DEG C, TEA (3.98mL, 28.5mmol, 5.0 equivalents) was added dropwise at -78 DEG C. Once addition is complete, the reaction mixture is slowly warmed to 0 DEG C and stirred for 30 minutes. The reaction is complete (analyzed by TLC (3:7 / EtOAc: petroleum ether, R f ~0.4, KMnO4) confirmed), the reaction mixture was diluted with DCM (10 mL) and washed with 10% aqueous citric acid (1×10 mL), water (1×10 mL) and brine (1×10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure below 30°C to afford crude product tert-butyl (3-oxopropyl)carbamate 6 (0.950 g, 4.90 mmol, 86% yield) as a light yellow liquid, which was used further without purification.

[1157] LCMS: (no mass peak observed). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm Mobile phase: A: 0.1% FA in H2O solution B: ACN, Flow rate: 1.5 mL / min. Rt (min):

[1158] 2.644; Area%-89.26.

[1159] Step 2: tert-Butyl (4-((3-((tert-Butoxycarbonyl)amino)propyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate

[1160]

[1161] Under nitrogen atmosphere, tert-butyl (3-oxopropyl)carbamate 6 (0.805 g, 4.65 mmol, 4 eq) and tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 4 (0.5 g, 1.161 mmol, 1 eq) were added together to a mixture of 1,2-dichloroethane (10 mL, 10 volumes) and DMF (5 mL, 5 volumes). Acetic acid (0.332 mL, 5.81 mmol, 5 eq) was added thereto, and the reaction mixture was stirred at room temperature for 1 hour. NaCNBH (0.292 g, 4.65 mmol, 4 eq) was added in portions, and the reaction mixture was stirred at room temperature for another 16 hours. The reaction was complete (by TLC analysis (100% EtOAc R f ~0.5) and LCMS confirmed), the reaction mixture was diluted with ice-cold water (15 mL), extracted with DCM (2×15 mL), washed with brine (25 mL), dried over anhydrous Na 2 SO 4 , filtered and concentrated to afford the crude product as a white solid, which was purified by preparative HPLC (Method: Instrument: SC-DC-ARD-05-045; Column number: XBridge C8-250-X-Select-C18-19×150 mm; 0.1% TFA in water / ACN; Rt: 9.6 min) and lyophilized to afford tert-butyl (4-((3-((tert-butoxycarbonyl)amino)propyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 7 (60 mg, 0.092 mmol, 7.89% yield) as a white solid.

[1162] LCMS: 588.4 (M+H). Method: Column: Bridge C8 (50×4.6 mm) 3.5 μm. Mobile phase: A: 0.1% TFA in H2O. B: 0.1% TFA in ACN. Flow rate: 1.5 mL / min. Rt (min): 1.911; Area % -89.75.

[1163] 1H-NMR (400MHz, DMSO-d6): δ10.97(s,1H),7.37(t,J=7.60Hz,1H),7.14(d,J=7.20Hz,1H),7.08(d,J=8.00Hz,1H),6.82-6.76(m,2H),5.10(dd,J =5.20,13.20Hz,1H),4.35(q,J=17.20Hz,2H),3.20-3.16(m,4H),2.96- 2.89(m,5H),2.04-2.01(m,1H),1.58-1.51(m,2H),1.58-1.51(m,23H).

[1164] Step 3: 3-(4-((4-aminobutyl)(3-aminopropyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1165]

[1166] To an ice-cold solution of tert-butyl (4-((3-((tert-butoxycarbonyl)amino)propyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 7 (60 mg, 0.102 mmol, 1 eq) in ethyl acetate (2 mL) was added HCl (4 M in EtOAc, 4 mL) and stirred at room temperature for 1.5 h. The reaction progress was monitored by LCMS. The reaction mixture was concentrated, washed with MTBE (2×5 mL) and lyophilized to afford 3-(4-((4-aminobutyl)(3-aminopropyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione 97 (20 mg, 0.051 mmol, 50.1% yield) as an off-white solid.

[1167] LCMS: 388.3 (M-2×HCl). Method: Column: Atlantis dC18 (50×4.6 mm) 5μm Mobile phase: A: 0.1% FA in H2O solution B: ACN, flow rate: 0.6 mL / min. Rt (min):

[1168] 2.218; Area%-99.01.

[1169] HPLC: 93.99%, Rt (min): 6.421. Method: Column: Atlantis dC18 (250×4.6) mm, 5 μm, mobile phase A: 0.1% TFA in H2O, mobile phase B: methanol, flow rate: 1.0 mL / min.

[1170] 1H-NMR(400MHz,DMSO-d6): δ11.00(s,1H),7.94(br s,6H),7.43(t,J=7.60Hz,1H),7.29-7.21(m,2H),5.13(dd,J=4.80,13.20Hz,1H),4.45(d,J=17.20Hz,1H),4.31(d,J=16.80Hz ,1H),3.76-3.20(m,4H),2.98-2.89(m,1H),2.81-2.76(m,4H),2.59(s,2H),2.08-2.02(m,1H),1.76-1.72(m,2H),1.53(s,4H).

[1171] Compound 102 in the form of a hydrochloride salt

[1172] 3-(4-((4-Aminobutyl)((1S,4S)-4-aminocyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1173]

[1174] Step 1: tert-Butyl ((1S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)carbamate

[1175]

[1176] 3-(4-Bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione 1 (0.5 g, 1.547 mmol, 1 eq), tert-butyl ((1S,4S)-4-aminocyclohexyl)carbamate 2 (0.995 g, 4.64 mmol, 3 eq) and NaOt-Bu (0.446 g, 4.64 mmol, 3 eq) were dissolved in DMF (2 mL) and degassed with nitrogen for 20 minutes. Pd-PEPPSI-IHept(Cl) (0.075 g, 0.077 mmol, 0.05 eq) was added under a nitrogen atmosphere and the reaction was heated at 110° C. for 4 h. The reaction was complete (by TLC analysis (100% EtOAc R f After 40 mmol / L (0.5 mmol / L) and LCMS confirmation), the reaction mixture was filtered through a bed of celite, washed with THF (10 mL), concentrated to a minimum volume (3 mL) and purified by reverse phase column chromatography ( The mixture was purified by HPLC-MS / ...

[1177] LCMS: 456.0 (M+H). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm Mobile phase: A: 0.1% FA in H2O B: ACN, Flow rate: 1.5 mL / min. Rt (min): 2.223; Area % -95.48.

[1178] 1 H-NMR (400MHz, DMSO-d6): δ11.04(s,1H),7.28(t,J=7.60Hz,1H),6.79(d,J=8.00Hz,1H),6.63(br s,1H),5.17-5.09(m,2H),4.21(q,J=17.20Hz,2H),3.47-3.42(m,2H),2.98-2.90(m ,1H),2.68-2.65(m,1H),2.08-2.03(m,1H),1.66(m,6H),0.54(m,3H),1.39(m,10H).

[1179] Step 2: tert-Butyl (tert-Butoxycarbonyl)(4-(((1S,4S)-4-((tert-Butoxycarbonyl)amino)cyclohexyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate

[1180]

[1181] To a solution of tert-butyl ((1S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)carbamate 3 (130 mg, 0.285 mmol, 1 eq), tert-butyl (tert-butoxycarbonyl)(4-oxobutyl)carbamate 4 (0.164 g, 0.569 mmol, 2 eq) in DCM (4 mL) and DMF (4 mL) at 0°C was added TFA (0.074 mL, 1.139 mmol, 4 eq) followed by sodium triacetoxyborohydride (0.241 g, 1.139 mmol, 4 eq). The reaction mixture was warmed to RT and stirred at the same temperature for 16 h. The reaction was complete (by TLC analysis (100% EtOAc Rf ~0.5) and LCMS confirmed), the reaction mixture was diluted with ice-cold water (15 mL), extracted with ethyl acetate (2×15 mL), washed with brine (25 mL), dried over anhydrous Na2SO4, filtered and concentrated. The resulting residue was purified by reverse phase column chromatography ( The purified product was purified using a 40 μm, 40 g column (C18, 40 μm, 40 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN aqueous mobile phase). Pure fractions from the column were lyophilized to afford tert-butyl (tert-butoxycarbonyl)(4-(((1S,4S)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 5 (74 mg, 0.140 mmol, 58% yield) as a white solid.

[1182] LCMS: 726.7 (MH). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm Mobile phase: A: 0.1% FA in H2O B: ACN, Flow rate: 1.5 mL / min. Rt (min): 2.912; Area % -99.75.

[1183] 1 H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.40(t,J=7.60Hz,1H),7.26(dd,J=8.00,16 .60Hz,2H),6.97(d,J=7.60Hz,1H),6.97(q,J=7.60Hz,1H),4.30(q,J=17.20Hz,2H),3. 56(m,1H),3.36-3.16(m,2H),3.08(m,1H),2.91-2.88(m,1H),2.01-1.99(m,1H),1.785 -1.809(m,2H),1.65-1.62(m,3H),1.53(m,8H),1.44-1.40(m,27H),1.24-1.21(m,3H).

[1184] Step 3: 3-(4-((4-aminobutyl)((1S,4S)-4-aminocyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1185]

[1186] To an ice-cold solution of tert-butyl (tert-butoxycarbonyl)(4-(((1S,4S)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 5 (72 mg, 0.099 mmol, 1 eq) in DCM (1 mL) was added HCl (4 M in EtOAc, 2.5 mL) and stirred at room temperature for 2 h. The reaction progress was monitored by LCMS. The reaction mixture was concentrated, washed with MTBE (2×5 mL) and lyophilized to afford 3-(4-((4-aminobutyl)((1S,4S)-4-aminocyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride 102 (49 mg, 0.095 mmol, 96% yield) as an off-white solid.

[1187] LCMS: 427.2 (M-HCl). Method: Column: XBridge C8 (50×4.6 mm), 3.5 μm. Mobile phase: A: 0.1% TFA in H2O; B: 0.1% TFA in ACN. Flow rate: 1.5 mL / min. Rt (min): 0.932; Area % -94.26.

[1188] HPLC: 95.86%, Rt (min): 1.363. Method: Column: X-Bridge C8 (50×4.6) mm, 3.5 μm, Mobile phase: A: 0.1% TFA in water, Mobile phase: B: ACN, Flow rate: 2.0 mL / min.

[1189] 1 H-NMR (400MHz, DMSO-d6): δ10.99(s,1H),8.23(s,3H),7.92(s,3H),7.44(d,J=7.20Hz,3H),5.10(dd,J=5.20,12.80Hz,1H),4.47-4. 29(m,2H),3.25(s,4H),2.73-2.70(m,1H),2.68-2.68(m,4H),2.05-2.02(m,1H),1.92-1.83(m,4H),1.92-1.83(m,6H),1.32(s,2H).

[1190] The hydrochloride form of compound 103

[1191] 3-(4-((4-Aminobutyl)((1R,4R)-4-aminocyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl

[1192]

[1193] Step 1: tert-Butyl ((1R,4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)carbamate

[1194]

[1195] 3-(4-Bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione 1 (0.5 g, 1.547 mmol, 1 eq), tert-butyl ((1R, 4R)-4-aminocyclohexyl)carbamate 2 (0.995 g, 4.64 mmol, 3 eq) and NaOt-Bu (0.446 g, 4.64 mmol, 3 eq) were dissolved in DMF (2 mL) and degassed with nitrogen for 20 minutes. Pd-PEPPSI-IHept(Cl) (0.075 g, 0.077 mmol, 0.05 eq) was added under a nitrogen atmosphere and the reaction was heated at 110 ° C for 4 h. After the reaction was complete (confirmed by UPLC analysis), the reaction mixture was filtered through celite, washed with THF (10 mL), concentrated to a minimum volume (3 mL) and purified by reverse phase column chromatography ( The mixture was purified by HPLC-MS / ...

[1196] LCMS: 401.1 (M-Boc). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm Mobile phase: A: 0.1% FA in H2O B: ACN, Flow rate: 1.5 mL / min. Rt (min): 2.200, Area % -99.01

[1197] 1H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),7.27(t,J=7.20Hz,1H),6.91(d,J= 7.20Hz,1H),6.80(t,J=8.00Hz,2H),5.29(d,J=6.80Hz,1H),5.12(q,J=4.80H z,1H),4.16(q,J=17.20Hz,2H),2.93-2.89(m,1H),2.70-2.60(m,2H),2.30- 2.20(m,2H),2.09-1.99(m,3H),1.80(m,2H),1.39(s,9H),1.32-1.24(m,4H).

[1198] Step 2: tert-Butyl (tert-Butoxycarbonyl)(4-(((1R,4R)-4-((tert-Butoxycarbonyl)amino)cyclohexyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate

[1199]

[1200] To a solution of tert-butyl ((1R,4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)carbamate 3 (70 mg, 0.153 mmol, 1 eq), tert-butyl (tert-butoxycarbonyl)(4-oxobutyl)carbamate 4 (88 mg, 0.307 mmol, 2 eq) in DCM (5 mL) and DMF (0.5 mL) at 0°C was added TFA (0.047 mL, 0.613 mmol, 4 eq) followed by sodium triacetoxyborohydride (130 mg, 0.613 mmol, 4 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was complete (by TLC analysis (100% EtOAc R f ~0.5) and LCMS confirmed), the reaction mixture was diluted with ice-cold water (15 mL), extracted with DCM (2×10 mL), washed with brine (25 mL), dried over Na 2 SO 4 , filtered and concentrated. The resulting residue was purified by reverse phase column chromatography ( The purified product was purified using a 40 μm column (C18, 100 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN aqueous solution mobile phase). Pure fractions from the column were lyophilized to afford tert-butyl (tert-butoxycarbonyl)(4-(((1R,4R)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 5 (40 mg, 0.055 mmol, 35.8% yield) as a white solid.

[1201] LCMS: 728.5 (M+H). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm Mobile phase: A: 0.1% FA in H2O B: ACN, Flow rate: 1.5 mL / min. Rt (min): 2.913; Area % -99.75.

[1202] Step 3: 3-(4-((4-aminobutyl)((1R,4R)-4-aminocyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl

[1203]

[1204] To an ice-cold solution of tert-butyl (tert-butoxycarbonyl)(4-(((1R,4R)-4-((tert-butoxycarbonyl)amino)cyclohexyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 5 (40 mg, 0.055 mmol, 1 eq) in DCM (1 mL) was added HCl (4 M in EtOAc, 1 mL, 4.0 mmol) and stirred at room temperature for 2 h. The reaction was complete (by TLC analysis (10% MeOH / DCM, R f ~0.1) confirmed), the reaction mixture was concentrated, washed with MTBE (2×5 mL) and further lyophilized to afford 3-(4-((4-aminobutyl)((1R,4R)-4-aminocyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl103 (13.2 mg, 0.026 mmol, 47.8% yield) as an off-white solid.

[1205] LCMS: 428.3 (M+H) Method: Column: XBridge C8 (50×4.6 mm) 3.5 μm Mobile phase: A: 0.1% TFA in H2O Mobile phase: B: 0.1% TFA in ACN, Flow rate: 1.5 mL / min, Rt (min): 0.906; Area% -95.74.

[1206] HPLC: 99.63%, Rt (min): 11.860. Method: Column: Atlantis dC18 (250×4.6) mm, 5 μm, Mobile phase A: 0.1% TFA in water, Mobile phase B: Methanol, Flow rate: 0.7 mL / min

[1207] 1H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.96(s,3H),7.80(s,3H),7.44(d,J= 7.20Hz,1H),7.34(t,J=8.80Hz,2H),5.12(q,J=5.20Hz,1H),4.38-4.23(m,2H) ,3.13(s,2H),2.98-2.89(m,3H),2.68(q,J=1.60Hz,2H),2.61(d,J=18.00Hz,2 H),2.04-1.91(m,3H),1.78(s,2H),1.62-1.50(m,4H),1.45(t,J=25.60Hz,4H).

[1208] Compound 104 in the form of a hydrochloride salt

[1209] 3-(4-((4-aminobutyl)-((1S,4S)-4-(aminomethyl)cyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1210]

[1211] Step 1: tert-Butyl (((1S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)methyl)carbamate

[1212]

[1213] To a solution of 3-(4-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione 1 (1.0 g, 3.09 mmol, 1.0 equiv) in DMF (5 mL, 5 volumes) was added tert-butyl (((1S,4S)-4-aminocyclohexyl)methyl)carbamate 2 (1.060 g, 4.64 mmol, 1.5 equiv) and sodium tert-butoxide (0.892 g, 9.28 mmol, 3.0 equiv) at room temperature and degassed by bubbling with nitrogen for a period of 15 minutes. To the reaction mixture was added Pd-PEPPSI-IHept(Cl) (0.151 g, 0.155 mmol, 0.05 equiv) under a nitrogen atmosphere, degassed for 5 minutes and the mixture was stirred at 85° C. overnight. The reaction was complete (by TLC and UPLC analysis (100% EtOAc, R f0.2g, 0.350mmol, 11.30% yield)。

[1214] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),7.25-7.29(m,1H),6.90-6.93(m,1H),6.80-6.87(m,1H),6.77-6.79(m,1H),5.11-5.21(m,2H),4.15 -4.30(m,2H),3.47-3.51(m,1H),2.92-2.99(m,3H),2.67-2.68(m,1H), 2.30-2.34(m,1H),2.02-2.08(m,1H),1.41-1.46(m,3H),1.38(m,15H).

[1215] LCMS: 415.8 (M-tBu), Rt (min): 2.324, area-89.8%.

[1216] HPLC: Rt (min): 5.496; Area -82.3%.

[1217] Step 2: tert-Butyl (tert-butoxycarbonyl)(4-oxobutyl)carbamate

[1218]

[1219] At-78 ℃, under nitrogen atmosphere, within 15 minutes, the solution of oxalyl chloride (1.815mL, 20.73mmol, 2.0 equivalents) in DCM (20mL) was dropwise added to the solution of DMSO (2.94mL, 41.5mmol, 4.0 equivalents) in DCM (5mL), and stirred 15 minutes at the same temperature.Within 15 minutes, dropwise add (tert-butoxycarbonyl) (4-hydroxybutyl) tert-butyl carbamate 4 (3.0g, 10.37mmol) at CH cl 2 solution in (5mL).After reaction mixture was stirred 30 minutes at-78 ℃, dropwise add triethylamine (8.67mL, 62.2mmol, 6.0 equivalents) at-78 ℃.Then, reaction mixture was slowly warmed to 0 ℃ and stirred 30 minutes.Reaction was completed (analyzed (20% EtOAc / petroleum ether, R by TLC)). f ~0.6, KMnO4) confirmed), the reaction mixture was diluted with DCM (40 mL) and washed with 10% aqueous citric acid (1×40 mL) and water (1×40 mL), dried over Na2SO4, filtered and concentrated under vacuum (below 30°C) to afford tert-butyl (tert-butoxycarbonyl)(4-oxobutyl)carbamate 5 (2.9 g, 9.99 mmol, crude product) as a light yellow liquid, which was used further without purification.

[1220] 1 H-NMR (400MHz, DMSO-d6): δ9.66(s,1H),3.47-3.50(m,1H),2.51-2.55(m,2H),2.42-2.44(m,2H),1.69-1.75(m,2H),1.43-1.45(s,18H),

[1221] Step 3: tert-Butyl (tert-butoxycarbonyl)(4-(((1S,4S)-4-(((tert-butoxycarbonyl)amino)methyl)cyclohexyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate

[1222]

[1223] To a mixture of tert-butyl (((1S,4S)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)methyl)carbamate 3 (0.2 g, 0.425 mmol, 1.0 equiv) and tert-butyl (tert-butoxycarbonyl)(4-oxobutyl)carbamate 5 (0.244 g, 0.850 mmol, 2.0 equiv) in CHCl (10 mL) and DMF (10.00 mL) was added trifluoroacetic acid (0.131 mL, 1.700 mmol, 4.0 equiv) and stirred at 0°C for 10 minutes. Sodium triacetoxyborohydride (0.360 g, 1.700 mmol, 4.0 equiv) was added portionwise at 0°C and the resulting colorless turbid material was stirred at room temperature for 16 h (monitored by UPLC). Reactant mixture is cooled to 0 DEG C and add additional amount of (tert-butoxycarbonyl) (4-oxobutyl) tert-butyl carbamate 5 (0.244g, 0.850mmol, 2.0 equivalents), trifluoroacetic acid (0.131mL, 1.700mmol, 4.0 equivalents), and stir 10 minutes.Then, at 0 DEG C, add sodium triacetoxyborohydride (0.360g, 1.700mmol, 4.0 equivalents), and gained colorless turbid material is at room temperature stirred 16 hours.Reaction is completed (by TLC analysis (8:2 / EtOAc: petroleum ether, R f After 4 hours (confirmed by 4% HCl, 0.6% HCl), the reaction mixture was quenched with ice-cold water (20 mL) and extracted with dichloromethane (2 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum to obtain the crude product as a light yellow liquid. The crude compound was purified by Isolera chromatography (Biotage R snap cartridge, KP-Sil, 100 g, 230-400 silica gel) using 60-70% ethyl acetate in petroleum ether. Fractions were collected and concentrated under vacuum to obtain a light yellow liquid.

[1224] The obtained compound was further purified by reverse phase chromatography (eluting with 75% ACN in 0.1% HCO2H) and the pure fractions were lyophilized to afford tert-butyl (tert-butoxycarbonyl)(4-(((1S,4S)-4-(((tert-butoxycarbonyl)amino)methyl)cyclohexyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 6 (0.11 g, 0.146 mmol, 34% yield) as a white solid.

[1225] 1H-NMR (400MHz, DMSO-d6): δ10.96(s,1H),7.40-7.43(m,1H),7.27-7.33(m,2H), 6.78-6.81(m,1H),5.07-5.12(m,1H),4.24-4.36(m,2H),3.33-3.39(m,2H),3.08 -3.12(m,3H),2.91-2.95(m,3H),2.60-2.61(m,2H),2.53-2.56(m,2H),1.98-2.0 1(m,1H),1.62-1.65(m,3H),1.52-1.53(m,6H),1.48(m,27H),1.22-1.24(m,2H).

[1226] LCMS: 741.4 (M + ), Rt(min):2.945, area%-89.8%.

[1227] HPLC: Rt (min): 5.816; Area % - 99.9%.

[1228] Step 4: 3-(4-((4-aminobutyl)((1S,4S)-4-(aminomethyl)cyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1229]

[1230] To a stirred solution of tert-butyl (tert-butoxycarbonyl)(4-(((1S,4S)-4-(((tert-butoxycarbonyl)amino)methyl)cyclohexyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 6 (0.1 g, 0.135 mmol, 1.0 equiv) in dichloromethane (10 mL) was added HCl (4 M in dioxane, 10 mL) at 0°C and then stirred at room temperature for 2 h. The reaction progress was monitored by LCMS. The reaction mixture was concentrated under vacuum at below 40°C to give an off-white solid. The solid obtained was dissolved in water (10 mL) and washed with MTBE (2 x 10 mL), the aqueous layer was lyophilized to afford 3-(4-((4-aminobutyl)((1S,4S)-4-(aminomethyl)cyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl (62.5 mg, 0.116 mmol, 86% yield) as an off-white solid.

[1231] 1H-NMR (400MHz, DMSO-d6): δ11.00(s,1H),7.89-7.98(m,6H),7.33-7.44(m,3H),5.10-5.13(m,1H),4.30-4.37(m,2H) ,3.43-3.49(m,2H),3.14-3.17(m,2H),2.84-2.96(m,6H),2.05(m,1H),1.86(m,2H),1.32-1.64(m,9H),1.24(m,2H).

[1232] LCMS: 442.3 (M+H). Method: Column: XBridge C8 (50×4.6 mm) 3.5 μm, Mobile phase: A: 0.1% TFA in H 2 O, Mobile phase: B: 0.1% TFA in acetonitrile, Flow rate: 1.5 mL / min.

[1233] HPLC: Method: Mobile phase A: 0.1% TFA in water, Mobile phase B: Methanol, Flow rate: 1.0 mL / min. Column: Atlantis dC18 (250×4.6) mm, 5 μm. RT: 6.488 min, Area: 95.427%.

[1234] Compound 105 in the form of a hydrochloride salt

[1235] 3-(4-((4-Aminobutyl)((1R,4R)-4-(aminomethyl)cyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl

[1236]

[1237] Step 1: tert-Butyl (((1R,4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)methyl)carbamate

[1238]

[1239] 3-(4-Bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione 1 (1.0 g, 3.09 mmol, 1 eq), tert-butyl (((1R,4R)-4-aminocyclohexyl)methyl)carbamate 2 (2.120 g, 9.28 mmol, 3 eq) and NaOt-Bu (0.892 g, 9.28 mmol, 3 eq) were dissolved in DMF (4 mL) and degassed with nitrogen for 20 minutes. Pd-PEPPSI-IHept(Cl) (0.151 g, 0.155 mmol, 0.05 eq) was added under a nitrogen atmosphere and the reaction was heated at 110 ° C for 4 h. After the reaction was complete (confirmed by UPLC analysis), the reaction mixture was filtered through celite, washed with EtOAc (5 mL), concentrated to a minimum volume (3 mL), and purified by reverse phase column chromatography ( The mixture was purified by HPLC-MS / ...

[1240] LCMS: 401.1 (M-Boc). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm Mobile phase: A: 0.1% FA in H2O B: ACN, Flow rate: 1.5 mL / min. Rt (min): 2.262, Area % -96.25

[1241] 1 H-NMR (400MHz, DMSO-d6): δ10.97(s,1H),7.26(t,J=7.60Hz,1H),6.91(d,J=7.60Hz,1 H),6.85-6.78(m,2H),5.28(d,J=8.40Hz,1H),5.12(dd,J=5.20,13.20Hz,1H),4.16(d, J=17.2Hz,2H),4.09(d,J=17.2Hz,1H),2.66(t,J=14.40Hz,1H),2.34-2.27(m,1H),2. 04-1.99(m,3H),1.73-1.70(m,2H),1.38(s,11H),1.29-1.14(m,2H),1.05-1.03(m,2H)

[1242] Step 2: tert-Butyl (tert-butoxycarbonyl)(4-(((1R,4R)-4-(((tert-butoxycarbonyl)amino)methyl)cyclohexyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate

[1243]

[1244] To a solution of tert-butyl (((1R,4R)-4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)cyclohexyl)methyl)carbamate 3 (90 mg, 0.191 mmol, 1 eq), tert-butyl (tert-butoxycarbonyl)(4-oxobutyl)carbamate 4 (110 mg, 0.383 mmol, 2 eq) in DCM (5 mL) and DMF (2 mL) was added TFA (0.059 mL, 0.765 mmol, 4 eq) and sodium triacetoxyborohydride (162 mg, 0.765 mmol, 4 eq) at 0° C. The resulting reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (confirmed by LCMS), the reaction mixture was diluted with ice-cold water (10 mL), extracted with DCM (2×10 mL), washed with brine (25 mL), dried over Na 2 SO 4 , filtered and concentrated. The resulting residue was purified by reverse phase column chromatography ( The purified product was purified using a 40 μm column (C18, 100 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN aqueous solution mobile phase). Pure fractions from the column were lyophilized to afford tert-butyl (tert-butoxycarbonyl)(4-(((1R,4R)-4-(((tert-butoxycarbonyl)amino)methyl)cyclohexyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 5 (20 mg, 0.025 mmol, 14.03% yield) as a white solid.

[1245] LCMS: 742.5 (M+H). Method: Column: XBridge C8 (50×4.6 mm) 3.5 μm, Mobile Phase: A: 0.1% TFA in H2O, Mobile Phase: B: 0.1% TFA in ACN, Flow Rate: 1.5 mL / min. Rt (min): 2.238; Area % -98.70.

[1246] 1H-NMR (400MHz, DMSO-d6): δ10.97 (s, 1H), 7.40 (t, J = 7.60Hz, 1H), 7.26 (q, J=8.00Hz,2H),6.83-6.82(m,1H),5.09(q,J=4.80Hz,1H),3.12(t,J=6.00H z,2H),3.00(m,2H),2.90(d,J=3.60Hz,2H),1.70(d,J=9.60Hz,4H),1.69- 1.38(m,4H),1.24(s,33H),1.19(d,J=9.20Hz,2H),0.87(q,J=6.00Hz,2H).

[1247] Step 3: 3-(4-((4-aminobutyl)((1R,4R)-4-(aminomethyl)cyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl

[1248]

[1249] To an ice-cold solution of tert-butyl (tert-butoxycarbonyl)(4-(((1R,4R)-4-(((tert-butoxycarbonyl)amino)methyl)cyclohexyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate (20 mg, 0.027 mmol, 1 eq) in DCM (1 mL) was added HCl (4 M in EtOAc, 1 mL, 4.0 mmol) and stirred at room temperature for 2 h. The product was analyzed by TLC (10% MeOH:DCM, R f After confirming the completion of the reaction, the reaction mixture was concentrated and lyophilized to obtain 3-(4-((4-aminobutyl)((1R,4R)-4-(aminomethyl)cyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl (7.5 mg, 0.014 mmol, 52.3% yield) as a light yellow solid.

[1250] LCMS: 428.3 (M+H) Method: Column: XBridge C8 (50×4.6 mm) 3.5 μm, Mobile phase: A: 0.1% TFA in H2O, Mobile phase: B: 0.1% TFA in ACN, Flow rate: 1.5 mL / min, Rt (min): 0.8 / 66; Area% -94.61.

[1251] HPLC: 96.75%, Rt(min): 2.674. Method: Method information: A: 0.1% TFA in H2O, B: methanol, flow rate: 1.0 mL / min, column: YMC Hydrosphere C18 (150×4.6 mm), 3 μm

[1252] 1 H-NMR (400MHz, DMSO-d6): δ11.00(s,1H),7.87-7.86(m,6H),7.44(s,1H),7.33(s, 2H),5.13-5.09(m,1H),4.31(q,J=16.40Hz,2H),3.13(s,2H),2.98(t,J=6.00Hz,1H ),2.89(d,J=5.20Hz,1H),2.71(s,2H),2.68(d,J=1.60Hz,3H),2.03(d,J=10.80Hz ,1H),1.7-1.9(m,4H),1.53(t,J=7.20Hz,5H),1.24-1.25(m,2H),1.12-1.0(m,2H).

[1253] Compound 106 in the form of a hydrochloride salt

[1254] 3-(4-((4-Aminobutyl)(((1S,4S)-4-aminocyclohexyl)methyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl

[1255]

[1256] Step 1: tert-Butyl ((1S,4S)-4-formylcyclohexyl)carbamate

[1257]

[1258] To a cooled solution of tert-butyl ((1S,4S)-4-(hydroxymethyl)cyclohexyl)carbamate (500 mg, 2.180 mmol, 1 eq) in DCM (11 mL) was added DMSO (3.7 mL), DIPEA (1.510 mL, 8.48 mmol, 3.89 eq) and pyridine sulfur trioxide (1.35 g, 8.48 mmol, 3.89 eq) in DMSO (1.5 mL) dropwise and stirred at 0° C. for 1 h. The reaction was complete (by TLC (40% ethyl acetate in petroleum ether, R fTo the 4-nitro-2-thiazolinone (50mg, 1.584mmol, 72.6% yield) of crude product ((1S, 4S)-4-formylcyclohexyl) tert-butyl carbamate 2A (360mg, 1.584mmol, 72.6% yield) of colorless liquid was added.

[1259] 1 H-NMR (400MHz, DMSO-d6): δ9.58(s,1H),8.23(s,1H),6.74(s,1H),3.65-3.56(m,2H),2.55-2.50(m,13H),1.41(s,3H).

[1260] Step 2a: tert-Butyl ((1S,4S)-4-(((4-((tert-Butoxycarbonyl)amino)butyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamate

[1261]

[1262] To a solution of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate Int-4 (200 mg, 0.465 mmol, 1 eq) and tert-butyl ((1S,4S)-4-formylcyclohexyl)carbamate 2A (528 mg, 2.323 mmol, 5 eq) in a mixture of DCE (2 mL) and DMF (0.5 mL) at 0° C. was added TFA (47.7 mg, 0.418 mmol, 0.9 eq) and Na(OAc)3BH (35.0 mg, 0.557 mmol, 1.5 eq) and allowed to stir at room temperature for 12 h. After completion of the reaction (confirmed by UPLC), the reaction mixture was quenched with ice water (25 mL) and extracted with DCM (2×10 mL). The combined organic layers were concentrated under vacuum to obtain the crude product. The crude product was purified by reverse phase chromatography ( 1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamic acid tert-butyl ester 2 (130 mg, 0.202 mmol, 43% yield) was purified using a column: general C18, 40 g snap, 0.1% formic acid in ACN, 20 mL / min, 70-75% ACN in H2O) and product fractions were lyophilized to afford tert-butyl ((1S,4S)-4-(((4-((tert-butoxycarbonyl)amino)butyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamate 2 as an off-white solid (130 mg, 0.202 mmol, 43% yield).

[1263] 1 H-NMR (400MHz, DMSO-d6): δ10.99(s,1H),7.37(t,J=7.60Hz,1H),7.19(d,J=7.20Hz,1H),7.1 1(d,J=8.00Hz,1H),6.78(t,J=5.20Hz,1H),6.64(d,J=6.80Hz,1H),5.17-5.08(m,1H),4.27( d,J=17.20Hz,1H),4.38(d,J=16.80Hz,1H),3.44(s,1H),3.13(dd,J=6.80,28.00Hz,4H),2.8 9(t,J=6.00Hz,3H),2.04(q,J=4.80Hz,1H),1.52(t,J=21.20Hz,4H),1.36(d,J=9.20Hz,28H).

[1264] LCMS: 642.4 (M+H), Rt (min): 2.992, Area %: 99.757.

[1265] HPLC: Rt (min): 4.691, Area %: 99.938.

[1266] Step 3a: 3-(4-((4-aminobutyl)(((1S,4S)-4-aminocyclohexyl)methyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl

[1267]

[1268] To an ice-cold solution of tert-butyl ((1S,4S)-4-(((4-((tert-butoxycarbonyl)amino)butyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamate (130 mg, 0.203 mmol, 1 equiv) in DCM (2.5 mL) at 0 °C was added HCl (4 M in ethyl acetate, 1.3 mL, 5.20 mmol) and stirred at room temperature for 2 h. After completion of the reaction (confirmed by LCMS), the reaction mixture was concentrated, washed with MTBE (10 mL) and dried to afford 3-(4-((4-aminobutyl)(((1S,4S)-4-aminocyclohexyl)methyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl 106 (85 mg, 0.165 mmol, 81% yield) as a light yellow solid.

[1269] 1H-NMR (400MHz, DMSO-d6): δ10.99(s,1H),8.10(d,J=35.60Hz,5H),7.43(s,1H),7.26(s,1H),5.12(q,J=5.20Hz,1H), 4.39(q,J=16.80Hz,2H), 3.16(d,J=26.00Hz,4H),2.97-2.88(m,1H),2.76(d,J=4.80Hz,2H),2.6 3(s,1H),2.51(t,J=1.60Hz,1H),2.05(t,J=5.20Hz,1H),1.68(s,12H).

[1270] LCMS: 442.4 (M+H), method: mobile phase: A: 0.1% TFA in H2O; mobile phase: B: 0.1% TFA in ACN, column: XBridge C8 (50×4.6 mm) 3.5 μm, flow rate: 1.5 mL / min, Rt (min): 1.027, area %: 99.640.

[1271] HPLC: Method: A: 0.1% TFA in H2O, B: ACN, Flow rate: 2.0 mL / min, Column: Xbridge C8 (50×4.6) mm, 3.5 μm Rt (min): 1.655, Area %: 99.741.

[1272] Compound 107 in the form of a hydrochloride salt

[1273] 3-(4-((4-Aminobutyl)(((1R,4R)-4-aminocyclohexyl)methyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl

[1274]

[1275] Step 1: tert-Butyl ((1R,4R)-4-formylcyclohexyl)carbamate 2A

[1276]

[1277] To a cooled solution of tert-butyl ((1R, 4R)-4-(hydroxymethyl)cyclohexyl)carbamate (500 mg, 2.180 mmol, 1 eq) in DCM (11 mL) was added DMSO (3.7 mL), DIPEA (1.510 mL, 8.48 mmol, 3.89 eq) followed by dropwise addition of a solution of pyridine sulfur trioxide (1.35 g, 8.48 mmol, 3.89 eq) in DMSO (3.7 mL). The mixture was stirred at 0 ° C for 15 minutes. The reaction was complete (by TLC (40% ethyl acetate in petroleum ether, R f To the 4-nitro-2-thiazolinone (500mg, 2.167mmol, 1.03mmol) of t-butyl ((1R, 4R)-4-formylcyclohexyl)carbamate 2A (500mg, 2.123mmol, 97% yield) of crude product ((1R, 4R)-4-formylcyclohexyl)carbamate) was added 1.5N HCl (20mL) and extracted with DCM (2 × 20mL). The organic layer merged was concentrated under vacuum to provide the crude product ((1R, 4R)-4-formylcyclohexyl)carbamic acid tert-butyl ester 2A (500mg, 2.123mmol, 97% yield) as a yellow solid.

[1278] 1 H-NMR (400MHz, DMSO-d6): δ9.55(d,J=0.80Hz,1H),6.75(d,J=7.60Hz,1H),3.16(s,1H),2.52(q, J=2.00Hz,3H),2.09(d,J=17.60Hz,2H),1.92-1.83(m,5H),1.38(s,1H),1.19(d,J=8.80Hz,5H).

[1279] LCMS: 172 (M-56), Rt (min): 2.587, Area %: 96.528.

[1280] Step 2: tert-Butyl ((1R,4R)-4-(((4-((tert-Butoxycarbonyl)amino)butyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamate

[1281]

[1282] To a solution of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate Int-4 (200 mg, 0.465 mmol, 1 eq) and tert-butyl ((1R,4R)-4-formylcyclohexyl)carbamate 2A (528 mg, 2.323 mmol, 5 eq) in a mixture of DCE (2 mL) and DMF (0.5 mL) at 0° C. was added TFA (47.7 mg, 0.418 mmol, 0.9 eq) and Na(OAc)3BH (35.0 mg, 0.557 mmol, 1.5 eq) and allowed to stir at room temperature for 12 h. After completion of the reaction (confirmed by UPLC), the reaction mixture was quenched with ice water (25 mL) and extracted with DCM (2×10 mL). The combined organic layers were concentrated under vacuum to provide a crude product. The crude product was purified by Isolera (column size: Biotage R snap cartridge, KP-Sil, 25 g, silica gel 230-400 mesh) using 50-60% ethyl acetate in petroleum ether to obtain tert-butyl ((1R,4R)-4-(((4-((tert-butoxycarbonyl)amino)butyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamate 2 (60 mg, 0.093 mmol, 19.97% yield) as a light yellow solid.

[1283] 1 H-NMR (400MHz, DMSO-d6): δ10.99(s,1H),7.37(t,J=7.60Hz,1H),7.18(d,J=7.20Hz,1H),7.09(d,J=8.00Hz, 1H),6.78(t,J=5.60Hz,1H),6.65(d,J=8.00Hz,1H),5.13-5.08(m,1H),4.38(d,J=16.80Hz,1H),4.28(d,J=16 .80Hz,1H),4.03(q,J=7.20Hz,1H),3.17(d,J=6.80Hz,3H),3.03(d,J=6.80Hz,2H),2.93-2.87(m,3H),2.56( t,J=2.00Hz,2H),2.02(t,J=9.20Hz,1H),1.71(d,J=8.80Hz,4H),1.36(d,J=2.00Hz,24H),1.04-0.90(m,4H),

[1284] LCMS: 642.4 (M+H), Rt (min): 2.900, Area %: 99.216.

[1285] HPLC: Rt (min): 4.591, Area %: 97.770.

[1286] Step 3: 3-(4-((4-aminobutyl)(((1R,4R)-4-aminocyclohexyl)methyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl

[1287]

[1288] To an ice-cold solution of tert-butyl ((1R,4R)-4-(((4-((tert-butoxycarbonyl)amino)butyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)methyl)cyclohexyl)carbamate 2 (60 mg, 0.093 mmol, 1 eq) in DCM (1.7 mL) was added HCl (4 M in ethyl acetate, 0.89 mL, 3.56 mmol, 5 vol) and stirred at room temperature for 6 h. After completion of the reaction (confirmed by UPLC), the reaction mixture was concentrated and washed with MTBE (10 mL) to afford the crude product, which was lyophilized to afford 3-(4-((4-aminobutyl)(((1R,4R)-4-aminocyclohexyl)methyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl (35 mg, 0.072 mmol, 77% yield) as a light yellow solid.

[1289] 1 H-NMR (400MHz, DMSO-d6): δ11.00(s,1H),7.98(t,J=31.60Hz,6H),7.40(t,J=7.60Hz ,1H),7.20(t,J=22.40Hz,2H),5.12(q,J=5.20Hz,1H),4.35(q,J=17.20Hz,2H),3.20 (s,2H),3.06(d,J=6.40Hz,2H),2.96-2.90(m,2H),2.88-2.68(m,2H),2.61(d,J=18. 40Hz, 2H), 2.04 (q, J = 5.20Hz, 1H), 1.21 ( q, J = 12.40Hz, 2H), 0.98 ( q, J = 12.40Hz, 2H).

[1290] LCMS: 441.0 (MH), method: mobile phase: A: 0.1% FA in H2O; mobile phase: B: ACN; column: Atlantis dC18 (50×4.6 mm) 5 μm; flow rate: 1.5 mL / min; Rt (min): 0.995, area %: 98.488.

[1291] HPLC: Method: Mobile phase A: 0.1% TFA in water; Mobile phase B: acetonitrile; Column: Atlantis dC18 (250×4.6) mm, 5 μm; Flow rate: 1.0 mL / min; Rt (min): 6.025, Area %: 97.654.

[1292] Hydrochloride forms of Compound 108 and Compound 109

[1293] 3-(4-((4-Aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[1294]

[1295] Step 1: 3-(4-(((1S,4S)-4-methylcyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione and 3-(4-(((1R,4R)-4-methylcyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione

[1296]

[1297] To a stirred solution of 3-(4-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione 1 (3 g, 11.57 mmol, 1.0 eq) in TFA (30 mL) was added sodium triacetoxyborohydride (12.26 g, 57.9 mmol, 5.0 eq) and stirred at -15 °C for 10 minutes, followed by a solution of 4-methylcyclohexane-1-one 2 (10.38 g, 93 mmol, 8.0 eq) in DCM (10 mL) and the reaction was stirred for 16 h. The solvent was removed under reduced pressure to obtain the crude product. The crude compound was purified by reverse phase column chromatography ( The product was purified by column: C18 40 μm, 350 g; flow rate: 60 mL / min; 0.1% HCOOH / ACN aqueous solution mobile phase). The desired fraction was distilled under reduced pressure to obtain 3-(4-((4-methylcyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (0.56 g, 13.62% yield) as an off-white solid as a mixture of cis and trans forms. The cis-trans mixture was separated by preparative HPLC (column: X-Select-C18-19 x 250 mm, mobile phase: 0.1% TFA in water / ACN, flow rate: 15 mL / min) to afford 3-(4-(((1S,4S)-4-methylcyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione 108A (0.29 g, 7.05% yield) as a pale yellow solid and 3-(4-(((1R,4R)-4-methylcyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione 109A (0.15 g, 3.65% yield) as a pale yellow solid.

[1298] Step 2: tert-Butyl (tert-butoxycarbonyl)(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)((1S,4S)-4-methylcyclohexyl)amino)butyl)carbamate

[1299]

[1300] To a stirred solution of 3-(4-(((1S,4S)-4-methylcyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione 108A (0.098 g, 0.276 mmol, 1.0 equiv) and tert-butyl (tert-butoxycarbonyl)(4-oxobutyl)carbamate (0.475 g, 1.654 mmol, 6.0 equiv) in DCM (5 mL) and DMF (2 mL) was added TFA (0.189 g, 1.654 mmol, 6.0 equiv) and sodium triacetoxyborohydride (0.497 g, 2.344 mmol, 8.5 equiv) at 0°C. The reaction mixture was warmed to RT and stirred for 16 h. After completion of the reaction (confirmed by UPLC), the reaction mixture was quenched by ice water (50 mL) and extracted with DCM (2×30 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse phase column chromatography ( The product was purified by HPLC-MS / MS (column: C18 40 μm, 120 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN aqueous mobile phase) to afford the desired product, tert-butyl (tert-butoxycarbonyl)(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)((1S,4S)-4-methylcyclohexyl)amino)butyl)carbamate (0.07 g, 39.7% yield) as an off-white solid.

[1301] 1 H-NMR (400MHz, DMSO-d6): δ10.98 (s, 1H), 7.42 (t, J = 7.60Hz, 1H), 7.34-7. 27(m,2H),5.13-5.08(m,1H),4.36-4.25(m,2H),3.41-3.38(m,2H),3.13- 3.10(m,3H),2.94-2.86(m,1H),2.68-2.67(m,2H),2.01-1.98(m,1H),1.7 2-1.64(m,3H),1.48-1.44(m,25H),1.38-1.23(m,3H),0.94-0.92(m,3H),

[1302] LCMS: 627.4 (M+H), Rt (min): 3.41, Area % -97.91. HPLC: Rt (min): 4.88, Area: 99.57%.

[1303] Step 3: 3-(4-((4-aminobutyl)((1S,4S)-4-methylcyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[1304]

[1305] To an ice-cold suspension of tert-butyl (tert-butoxycarbonyl)(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)((1S,4S)-4-methylcyclohexyl)amino)butyl)carbamate 5 (0.07 g, 0.112 mmol, 1.0 equiv) in DCM (3 mL) was added HCl (4 M in EtOAc, 1 mL) at 0°C. The reaction mixture was stirred at RT for 2 h. The reaction mixture was concentrated and washed with methyl tert-butyl ether (5 mL). The solvent was decanted and concentrated under reduced pressure to give the crude product. The crude product was lyophilized to afford 3-(4-((4-aminobutyl)((1S,4S)-4-methylcyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl 108 (0.06 g, 106% yield) as an off-white solid.

[1306] 1 H-NMR (400MHz, DMSO-d6): δ10.99(s,1H),7.81(br s,4H),7.34-7.44(m,2H),5.12(t,J=8.40Hz,1H),4.31-4.35(m,2H),3.13(br s,2H),2.92(s,1H),2.60-2.71(m,3H),2.01-2.05(m,1H),1.69-1.77(m,4H),1.57-1.63(m,8H),1.24-1.31(m,2H),0.93-0.94(m,3H).

[1307] LCMS: 427.3 (M+H). Method: Mobile phase A: 0.1% TFA in H2O. Mobile phase B: acetonitrile. Flow rate: 1.5 mL / min. Column: XBridge C8 (50 × 4.6 mm), 3.5 μm. Rt (min): 1.340; Area % -99.13.

[1308] HPLC: Method: Mobile phase A: 0.1% TFA in H2O. Mobile phase B: acetonitrile. Flow rate: 2.0 mL / min. Column: Xbridge C8 (50 x 4.6 mm, 3.5 μm). Rt (min): 2.38; Area %: 98.95.

[1309] Step 2A: tert-Butyl (tert-butoxycarbonyl)(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)((1R,4R)-4-methylcyclohexyl)amino)butyl)carbamate

[1310]

[1311] To a stirred solution of 3-(4-(((1R,4R)-4-methylcyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione 109A (0.1 g, 0.281 mmol, 1.0 equiv) and tert-butyl (tert-butoxycarbonyl)(4-oxobutyl)carbamate (0.162 g, 0.563 mmol, 2.0 equiv) in DMF (5 mL):DCM (5 mL) was added TFA (0.160 g, 1.407 mmol, 4.0 equiv). After 10 minutes, sodium triacetoxyborohydride (0.239 g, 1.125 mmol, 4.0 equiv) was added portionwise to the mixture at 0°C. The reaction mixture was stirred at room temperature for 4 hours. The reaction was monitored for completion by UPLC, which indicated 39% SM. The reaction mixture is cooled at 0 DEG C and tert-butyl (tert-butoxycarbonyl) (4-oxobutyl) carbamate (0.485g, 1.688mmol, 6.0 equivalents) is added, followed by TFA (0.160g, 1.407mmol, 5.0 equivalents). After 10 minutes, sodium triacetoxyborohydride (0.507g, 2.391mmol, 8.5 equivalents) is added, and the reaction mixture is stirred at room temperature for 16 hours. The reaction is monitored by UPLC to complete. The reaction mixture is quenched by frozen water (20mL) and extracted with DCM (3 × 20mL). The organic layer merged is dried over sodium sulfate, filtered and concentrated under reduced pressure, to obtain crude product. The crude compound is purified by flash column chromatography (silica gel, 230-400 mesh size) using ethyl acetate / petroleum ether (50%) as eluent, to obtain the desired product as off-white solid. The product was repurified by reverse phase column chromatography (Grace column: C18 40 μm, 50 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN aqueous mobile phase) to obtain tert-butyl (tert-butoxycarbonyl)(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)((1R,4R)-4-methylcyclohexyl)amino)butyl)carbamate 5A (0.04 g, 20.87% yield) as an off-white solid.

[1312] 1H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.42-7.38(m,1H),7.30-7.23(m, 2H),5.11-5.07(m,1H),4.34-4.23(m,2H),3.12-3.11(m,2H),2.86-3.01(m, 2H),2.65-2.70(m,1H),2.01-1.98(m,1H),1.69-1.66(m,4H),1.53-1.46(m, 5H),1.37(s,18H),1.22-1.33(m,3H),0.99-0.91(m,2H),0.85-0.83(m,3H).

[1313] LCMS: 627.3 (MH), Rt (min): 2.27, Area%-92.32.

[1314] Step 3A: 3-(4-((4-aminobutyl)((1R,4R)-4-methylcyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[1315]

[1316] To an ice-cold solution of tert-butyl (tert-butoxycarbonyl)(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)((1R,4R)-4-methylcyclohexyl)amino)butyl)carbamate 5A (0.04 g, 0.064 mmol, 1.0 equiv) in DCM (3 mL) was added HCl (4 M in EtOAc, 1 mL) at 0°C. The reaction mixture was stirred at room temperature for 2 h. The reaction was complete (by TLC analysis (100% EtOAc R f After 4 hours of drying (10 min, 15 min), the reaction mixture was vacuum distilled to obtain a crude product. The crude product was washed with MTBE (5 mL), the solvent was decanted, dried under reduced pressure and lyophilized to afford the desired product 109, 3-(4-((4-aminobutyl)((1R,4R)-4-methylcyclohexyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (0.03 g, 101% yield), as an off-white solid.

[1317] 1H-NMR (400MHz, DMSO-d6): δ11.01(s,1H),7.99(brs,3H),7.37-7.51(m,3H),5.10-5.15(m,1H),4.44-4.63(m,2H),3.3 0(s,2H),3.01(s,3H),2.92-2.98(m,2H),2.60-2.78(m,3H),2.45-2.50(m,1H),2.06-2.08(m,1H),1.66-1.67(m,4H).

[1318] LCMS: 345.1 (M+H). Method: Mobile phase A: 0.1% TFA in H2O. Mobile phase B: acetonitrile. Flow rate: 1.5 mL / min. Column: XBridge C8 (50 × 4.6 mm), 3.5 μm. Rt (min): 1.351; Area % -99.19.

[1319] HPLC: Method: Mobile phase A: 0.1% TFA in H2O. Mobile phase B: acetonitrile. Flow rate: 2.0 mL / min. Column: Xbridge C8 (50 x 4.6 mm, 3.5 μm). Rt (min): 2.418; Area %: -99.64.

[1320] Compound 115 in the form of a hydrochloride salt

[1321]

[1322] To a solution of compound 10 (120 mg, 0.2 mmol, 1.0 equiv) in DCM (6.0 mL) was added piperidine (338 mg, 4.0 mmol, 20.0 equiv). The mixture was stirred at room temperature for 17 hours. The reaction mixture was then concentrated under reduced pressure and purified by P-HPLC (acetonitrile / HO: 20%-45%) to afford compound 115 (11.4 mg, 13%) as a white solid.

[1323] TLC: DCM / methanol = 10 / 1

[1324] R f (Compound 10) = 0.9

[1325] R f (Compound 115) = 0.1

[1326] LCMS: [C 21 H 26 N4O3] + Calculated value: 383, found value: 383.

[1327] 1H NMR (400MHz, D2O): δ7.72(d,J=7.3Hz,1H),7.66(d,J=7.9Hz,1H),7.60(t,J=7. 7Hz, 1H), 5.06 (dd, J=13.3, 4.9Hz, 1H), 4.53 (q, J=17.5Hz, 2H), 4.27 (s, 2H), 3. 60(s,2H),3.50-3.35(m,2H),2.97-2.62(m,2H),2.47-2.30(m,1H),2.25-2.10 (m,1H),1.37-1.20(m,2H),1.13(dd,J=14.5,7.2Hz,2H),0.65(t,J=7.2Hz,3H).

[1328] Compound 116 in the form of a hydrochloride salt

[1329]

[1330] To a solution of compound 4 (20 mg, 0.04 mmol, 1.0 equiv) in MeOH (1 mL) was added HCl (0.2 mL, 2 N, 0.4 mmol, 10.0 equiv). The mixture was stirred at room temperature for 24 hours and concentrated under reduced pressure. The residue was purified by Biotage (C18, acetonitrile / HO: 30%-60%) to afford compound 116 (14.0 mg, 74%) as a white solid.

[1331] TLC: DCM / methanol = 10 / 1

[1332] R f (Compound 4) = 0.5

[1333] R f (Compound 116) = 0.1

[1334] LCMS: [C 21 H 26 N4O3] + Calculated value: 383, found value: 383.

[1335] 1 H NMR (400MHz, D2O): δ8.30-8.20(m,2H),7.42-7.25(m,3H),4.98(d,J=12.6Hz,1H),4.38(q,J=17.2Hz,2H),3.67(s ,2H),3.11-2.98(m,2H),2.81-2.62(m,4H),2.42-2.22(m,1H),2.15-2.01(m,1H),1.53(s,3H),1.50-1.28(m,4H).

[1336] The hydrochloride form of compound 123

[1337] 3-(4-((4-amino-3,3-dimethylbutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[1338]

[1339] Step 1: 4-Methoxy-2,2-dimethyl-4-oxobutanoic acid

[1340]

[1341] To an ice-cold solution of 2,2-dimethylsuccinic acid 1 (100 g, 684 mmol, 1 eq) in methanol (1500 mL, 15 vol) was added concentrated H2SO4 (10 mL) dropwise and stirred at room temperature for 16 h. The reaction was complete (by TLC analysis (40% EtOAc in petroleum ether, R f ~0.5, KMnO4) confirmed), the reaction mixture was concentrated under vacuum at 30 ℃. The resulting residue was poured into an ice-cold solution of saturated NaHCO3 solution (1000mL) and washed with EtOAc (2×500mL). The separated aqueous layer was further acidified to pH~2 with HCl (6N) aqueous solution and extracted with EtOAc (2×800mL). The combined organic extracts were washed with brine (1×600mL), dried over Na2SO4, filtered and concentrated to obtain 4-methoxy-2,2-dimethyl-4-oxobutanoic acid 2 (40g, 235mmol, 34.3% yield) as a colorless liquid.

[1342] 1 H-NMR (400MHz, DMSO-d6): δ12.17(s,1H),3.57(s,3H),2.53(s,2H),1.18(s,6H).

[1343] Step 2: Methyl 4-amino-3,3-dimethyl-4-oxobutanoate

[1344]

[1345] To an ice-cold solution of 4-methoxy-2,2-dimethyl-4-oxobutanoic acid 2 (25 g, 156 mmol, 1 eq) in DCM (250 mL, 10 vol) was added thionyl chloride (111 g, 937 mmol, 6 eq) dropwise and stirred at room temperature for 3 h. The reaction was complete (by TLC analysis (20% EtOAc in petroleum ether, R fAfter 4% methyl ester (confirmed to be methyl ester), the reaction mixture was distilled under vacuum and completely dried under reduced pressure.

[1346] The gained acyl chloride mixture is dissolved in THF (250mL), and it is added dropwise to pre-cooled NH solution (0.5M THF solution, 375mL) at -78 ℃.The reaction mixture is warmed to room temperature and stirred for 30 minutes. The reaction is completed (by TLC analysis (60% EtOAc petroleum ether solution, R f After 4 hours, 4-amino-3,3-dimethyl-4-oxobutanoic acid methyl ester 3 (8 g, 50.3 mmol, 32.2% yield) was obtained.

[1347] 1 H-NMR (400MHz, DMSO-d6): δ6.75-7.07(m,2H),3.54(s,3H),2.51(s,2H),1.15(s,6H)

[1348] Step 3: 4-Amino-3,3-dimethylbutan-1-ol

[1349]

[1350] A solution of methyl 4-amino-3,3-dimethyl-4-oxobutanoate 3 (13 g, 82 mmol, 1 eq) in THF (100 mL, 8 vol) was treated dropwise with LiAlH4 (2 M in THF, 82 mL, 163 mmol, 2 eq) at 0°C over a period of 30 min. Once the addition was complete, the reaction mixture was allowed to warm to room temperature, slowly heated to 65°C and stirred for 2 h. The reaction was complete (by TLC analysis (100% EtOAc, R f After 4-amino-3,3-dimethylbutan-1-ol (7.8 g, 64.0 mmol, 78% yield) was obtained, the reaction mixture was cooled to 0 ° C. and quenched slowly with saturated Na2SO4 solution (30 mL). The precipitated inorganic salts were removed by filtration through a celite pad and washed with EtOAc (1×100 mL). The combined filtrates were dried over Na2SO4, filtered and concentrated to afford 4-amino-3,3-dimethylbutan-1-ol 4 (7.8 g, 64.0 mmol, 78% yield) as a light brown liquid, which was used without further purification.

[1351] 1H-NMR (400MHz, DMSO-d6): δ3.40-3.43(m,2H),2.30(s,2H),1.35-1.38(m,2H),0.80(s,6H).

[1352] Step 4: tert-Butyl (4-hydroxy-2,2-dimethylbutyl)carbamate

[1353]

[1354] To a solution of 4-amino-3,3-dimethylbutan-1-ol 4 (7.5 g, 57.2 mmol, 1 eq) in THF (150 mL, 20 vol) and water (40 mL, 5 vol) was added sodium bicarbonate (7.2 g, 86 mmol, 1.5 eq) and (Boc) O (14 mL, 57.2 mmol, 1 eq) dropwise at room temperature. The resulting reaction mixture was stirred at room temperature for 16 h. The reaction was complete (by TLC analysis (100% EtOAc, R f The product was concentrated under reduced pressure (100 mL) and the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.Then the mixture was stirred for 2 hours.

[1355] 1 H-NMR (400MHz, DMSO-d6): δ6.70(s,1H),4.28(t,J=6.40Hz,1H),3.41-3.47(m,2H),2.76(d,J=8.80Hz,2H),1.43(s,9H),0.83(s,6H).

[1356] Step 5: tert-Butyl 2-hydroxy-4,4-dimethylpyrrolidine-1-carboxylate

[1357]

[1358] At -78 ℃, to a solution of oxalyl chloride (0.5mL, 5.52mmol, 1.2 equivalents) in DCM (5mL) was added dropwise DMSO (1mL, 14.08mmol), and stirred at the same temperature for 15 minutes. Over a period of 15 minutes, a solution of tert-butyl (4-hydroxy-2,2-dimethylbutyl) carbamate (1.0g, 4.6mmol, 1 equivalent) in DCM (3mL) was added dropwise thereto and stirred at -78 ℃ for 30 minutes. Now, at -78 ℃, N, N-diisopropylethylamine (2.4mL, 13.81mmol, 3 equivalents) was added dropwise to the mixture and slowly warmed to 0 ℃. After stirring at 0 ℃ for 30 minutes, the mixture was analyzed by TLC (20% EtOAc in petroleum ether, R f The reaction mixture was diluted with DCM (20 mL) and washed with 10% aqueous citric acid (1×50 mL), water (1×30 mL) and brine (1×30 mL), dried over Na 2 SO 4 , filtered and concentrated under vacuum below 30° C. to afford tert-butyl 2-hydroxy-4,4-dimethylpyrrolidine-1-carboxylate (0.95 g, crude product) as a light yellow liquid, which was used further without purification.

[1359] GCMS: mass found MS: 215.1; Rt(min): 3.005. Method: HP5_SCAN350_MS.amx

[1360] Step 6: tert-Butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-2,2-dimethylbutyl)carbamate

[1361]

[1362] To a stirred mixture of 3-(4-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione (1.0 g, 3.86 mmol, 1.0 eq) and tert-butyl 2-hydroxy-4,4-dimethylpyrrolidine-1-carboxylate (0.996 g, 4.63 mmol, 1.2 eq) in CHCl (10 mL) and DMF (10.00 mL) was added trifluoroacetic acid (1.189 mL, 15.43 mmol, 4.0 eq) and stirred at 0°C for 10 minutes. To this mixture was added sodium triacetoxyborohydride (3.27 g, 15.43 mmol, 4.0 eq) in portions at 0°C, and the resulting colorless turbid material was stirred at room temperature for 16 hours. The reaction was complete (by TLC and LCMS analysis (100% EtOAc, R f7-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-2,2-dimethylbutyl)carbamic acid tert-butyl ester 8 (0.6 g, 1.049 mmol, 27.2% yield) was added to the crude product of 4- ((2- (2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-2,2-dimethylbutyl) carbamate (0.6 g, 1.049 mmol, 27.2% yield) using a petroleum ether solution of 75-85% ethyl acetate.

[1363] 1 H-NMR (400MHz, DMSO-d6): δ11.02(s,1H),7.29(t,J=10.40Hz,1H),6.93(d,J=10 .00Hz,1H),6.82-6.86(m,1H),6.75(d,J=10.80Hz,1H),5.49(s,1H),5.08-5.15( m,1H),4.08-4.25(m,2H),3.13-3.14(m,2H),2.73-2.91(m,3H),2.65(m,1H),2. 30-2.36(m,1H),2.01-2.08(m,1H),1.45-1.50(m,2H),1.38(s,9H),0.94(s,6H). LC-MS: 403.1 (M-tBu), Rt (min): 2.302, area% -80.229.

[1364] Step 7: tert-Butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)-2,2-dimethylbutyl)carbamate

[1365]

[1366] To a stirred mixture of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-2,2-dimethylbutyl)carbamate 8 (0.3 g, 0.654 mmol, 1.0 eq) and valeraldehyde 9 (0.564 g, 6.54 mmol, 10.0 eq) in CHCl (5 mL) and DMF (5.00 mL) was added trifluoroacetic acid (0.202 mL, 2.62 mmol, 4.0 eq) and stirred at 0°C for 10 minutes. Sodium triacetoxyborohydride (0.555 g, 2.62 mmol, 4.0 eq) was added portionwise at 0°C and the resulting colorless, turbid material was stirred at room temperature for 16 h. The reaction was complete (by TLC and LCMS analysis (100% EtOAc, R f 0.145mmol, 22.21% yield)。

[1367] 1 H-NMR (400MHz, DMSO-d6): δ10.99(s,1H),7.34-7.39(m,1H),7.16-7.18(m,1H),6.82-7.05(m,2H),5.09-5.13(m,1H),4.25-4. 42(m,2H),3.16-3.34(m,4H),2.87-2.92(m,1H),2.75-2.77(m,3H),2.02(m,1H),1.36(m,9H),1.25(s,9H),0.80-0.83(m,9H).

[1368] LCMS: 529.3 (M+H), Rt (min): 2.715, Area% -99.791.

[1369] Step 8: 3-(4-((4-amino-3,3-dimethylbutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride

[1370]

[1371] To a stirred suspension of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)-2,2-dimethylbutyl)carbamate 10 (80 mg, 0.151 mmol) in dichloromethane (2 mL) at 0°C was added HCl (4 M in ethyl acetate, 5 mL, 165 mmol) and then stirred at room temperature for 16 h. The reaction was complete (by TLC analysis (100% EtOAc, R f After 4 hours, the reaction mixture was concentrated under vacuum. The obtained solid was dissolved in water (10 mL) and washed with MTBE (10 mL). The aqueous layer was lyophilized to obtain 3-(4-((4-amino-3,3-dimethylbutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl 123 (70 mg, 0.145 mmol, 96% yield) as a light brown solid.

[1372] 1 H-NMR (400MHz, DMSO-d6): δ11.00(s,1H),7.87(s,3H),7.16-7.43(m,3H),5.11(s,1H),4.33-4.35(m,2H),3.22 (m,4H),2.90(m,1H),2.63(m,3H),2.50(m,3H),2.10(m,1H),1.43(m,3H),1.24(m,4H),0.94(m,5H),0.83(m,3H)

[1373] LCMS: 429.3 (M+H). Method: Column: X-BRIDGE C8 (50×4.6) 3.5 μm, Mobile phase: A: 0.1% TFA in H 2 O, B: 0.1% TFA in acetonitrile, Flow rate: 1.5 mL / min.

[1374] HPLC: Method: Mobile phase: A: 10 mM ammonium bicarbonate in water, Mobile phase: B: acetonitrile, Flow rate: 1.0 mL / min, Column: X-Bridge C8 (50×4.6) mm, 3.5 μm, RT: 5.333 min, Area: 96.027%.

[1375] Compound 124 in the form of a hydrochloride salt

[1376] 3-(4-((4-amino-3,3-dimethylbutyl)(4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1377]

[1378] Step 1: tert-Butyl (4-((tert-Butyldimethylsilyl)oxy)butyl)carbamate

[1379]

[1380] To a solution of tert-butyl (4-hydroxybutyl)carbamate (5.0 g, 26.4 mmol, 1.0 eq) in dichloromethane (100 mL) was added imidazole (4.5 g, 66.0 mmol, 2.5 eq) and TBS-Cl (4.78 g, 31.7 mmol, 1.5 eq) at 0° C. and allowed to stir at room temperature for 16 hours. The reaction was complete (by TLC (20% EtOAc / petroleum ether, R f To the 4- ((tert-butyldimethylsilyl)oxy)butyl)carbamic acid tert-butyl ester (8.0 g, 26.3 mmol, 99% yield) of 4- ((tert-butyldimethylsilyl)oxy)butyl)carbamate was added 1% ethyl acetate (50 mL) and 1% ethyl acetate (50 mL). The mixture was stirred for 2 hours at 37 ℃ for 3 hours at 4 ℃ for 10 minutes. After evaporation, 1% ethyl acetate (50 mL) was added 1% ethyl acetate (50 mL) and 1% ethyl acetate (50 mL). The mixture was stirred for 2 hours at 3 ℃ for 3 hours at 4 ℃ for 3 hours. After evaporation, 1% ethyl acetate (50 mL) was added 1% ethyl acetate (50 mL) and 1% ethyl acetate (50 mL). The mixture was stirred for 2 hours at 3 ℃ for 3 hours at 4 ℃ for 3 hours.

[1381] 1 H-NMR (400MHz, DMSO-d6): δ6.77(s,1H),3.56(t,2H),2.90(t,2H),1.16-1.20(m,4H),1.07(s,9H),0.86(s,9H),0.03(s,6H).

[1382] Step 2: tert-Butyl (tert-butoxycarbonyl)(4-((tert-butyldimethylsilyl)oxy)butyl)carbamate

[1383]

[1384] At 0 ℃, to a stirred solution of tert-butyl (4-((tert-butyldimethylsilyl)oxy)butyl)carbamate 2 (8.0 g, 26.4 mmol, 1.0 equivalent) in THF (80 mL), n-BuLi (2.5 M hexane solution, 12.65 mL, 31.6 mmol, 1.2 equivalents) was added, and stirred at 0 ℃ for 15 minutes. To the reaction mixture was added (Boc) at the same temperature a solution of O (7.34 mL, 31.6 mmol, 1.2 equivalents) in THF (20 mL). The resulting reaction mixture was warmed to room temperature and stirred for 1 h. The reaction was complete (by TLC analysis (10% EtOAc / petroleum ether, R f 7, KMnO 4 ) was added to the mixture, and the reaction mixture was quenched with water (50 mL) and extracted with dichloromethane (2×100 mL). The combined organic layers were dried over Na 2 SO 4 and concentrated under reduced pressure to afford the crude product tert-butyl (tert-butoxycarbonyl)(4-((tert-butyldimethylsilyl)oxy)butyl)carbamate 3 (11.0 g, 27.0 mmol, 102% yield) as a light yellow liquid, which was used further without purification.

[1385] 1 H-NMR (400MHz, DMSO-d6): δ3.58(t,2H),3.48(t,2H),1.16-1.20(m,4H),1.07(s,18H),0.86(s,9H),0.03(s,6H).

[1386] Step 3: tert-Butyl (tert-butoxycarbonyl)(4-hydroxybutyl)carbamate

[1387]

[1388] To a stirred solution of tert-butyl (tert-butoxycarbonyl)(4-((tert-butyldimethylsilyl)oxy)butyl)carbamate 3 (10 g, 24.77 mmol, 1.0 eq) in THF (160 mL) was added TBAF (1 M in THF, 24.77 mL, 24.77 mmol, 1.5 eq) at room temperature and stirred at the same temperature overnight. The reaction was complete (by TLC analysis (20% EtOAc / petroleum ether, R f53mmol, 83% productive rate).By the 1% tert-butyloxycarbonyl (4-hydroxybutyl) carbamate 4 (6g, 20.53mmol, 83% productive rate) of the petroleum ether solution of using 15-20% EtOAc, 4-hydroxybutyl carbamate 4 (6g, 20.53mmol, 83% productive rate).

[1389] 1 H-NMR (400MHz, DMSO-d6): δ4.40(t,J=6.80Hz,1H),3.41-3.48(m,2H),3.33-3.39(m,2H),1.44(m,4H),1.27(s,18H)

[1390] Step 4: tert-Butyl (tert-butoxycarbonyl)(4-oxobutyl)carbamate

[1391]

[1392] At-78 ℃, to oxalyl chloride (1.8mL, 20.73mmol, 2.0 equivalents) and CH cl 2 (30mL) solution adds DMSO (2.95mL, 41.5mmol, 4.0 equivalents) at CH cl 2 (5mL) in solution, and stirred 15 minutes at the same temperature.At-78 ℃, to this mixture, add (tert-butoxycarbonyl) (4-hydroxybutyl) tert-butyl carbamate 4 (3g, 10.37mmol, 1.0 equivalents) at CH cl 2 (10mL) in solution.After stirring reaction mixture at-78 ℃ for 30 minutes, at-78 ℃, drip triethylamine (9mL, 62.2mmol, 6.0 equivalents).Then, reaction mixture is slowly warmed to 0 ℃ and stirred 30 minutes.Reaction is completed (analyze (45% EtOAc / petroleum ether, R by TLC)). f After 40 ℃ of drying (10min, 10min, 10min of 4-nitropropanediol, 0.1mmol, 0.1mmol), the reaction mixture was diluted with DCM (20mL) and washed with 10% aqueous citric acid solution (1×100mL) and water (1×100mL), dried over Na 2 SO 4 and concentrated under reduced pressure (below 30° C.). The crude product was co-distilled with toluene to give tert-butyl (tert-butoxycarbonyl)(4-oxobutyl)carbamate 5 (3.2 g, 11.14 mmol, 107% yield) as a yellow liquid, which was used further without purification.

[1393] 1H-NMR (400MHz, DMSO-d6): δ9.65 (s, 1H), 3.48 (t, J = 9.20Hz, 2H), 2.41-2.50 (m, 2H), 1.70-2.30 (m, 2H), 1.44 (s, 18H),

[1394] Step 5: tert-Butyl (tert-butoxycarbonyl)(4-((4-((tert-butoxycarbonyl)amino)-3,3-dimethylbutyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate

[1395]

[1396] To a stirred mixture of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-2,2-dimethylbutyl)carbamate 8 (0.3 g, 0.654 mmol, 1.0 equiv) and tert-butyl (tert-butoxycarbonyl)(4-oxobutyl)carbamate 5 (0.376 g, 1.308 mmol, 2.0 equiv) in CHCl (5 mL) and DMF (5.00 mL) was added trifluoroacetic acid (0.202 mL, 2.62 mmol, 4.0 equiv) and stirred at 0° C. for 10 minutes. To this mixture was added sodium triacetoxyborohydride (0.555 g, 2.62 mmol, 4.0 equiv) portionwise at 0° C. and the resulting colorless turbid material was stirred at room temperature for 16 h. The reaction was complete (by TLC analysis (100% EtOAc, R f 75mg, 0.100mmol, 15.28% yield)。

[1397] 1H-NMR (400MHz, DMSO-d6): δ10.95(s,1H),7.35-7.39(m,1H),7.19-7.21(m ,1H),7.05-7.07(m,1H),6.79-6.81(m,1H),5.10-5.14(m,1H),4.26-4.41( m,2H),3.43-3.46(m,2H),3.17-3.19(m,4H),2.89-2.93(m,1H),2.68-2.75 (m,2H),2.00-2.02(m,2H),1.29-1.49(m,16H),1.24(s,18H),0.80(s,6H).

[1398] LCMS: 730.4 (M+H). Method: Atlantis dC18 (50×4.6 mm) 5 μm, mobile phase: A: 0.1% FA in H 2 O, mobile phase: B: acetonitrile, flow rate: 1.5 mL / min.

[1399] HPLC: Method: Mobile phase A: 0.1% TFA in water, Mobile phase B: acetonitrile, Flow rate: 2.0 mL / min. Column: X-Bridge C8 (50×4.6) mm, 3.5 μm. RT: 5.166 min, Area: 97.332%

[1400] Step 6: 3-(4-((4-amino-3,3-dimethylbutyl)(4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1401]

[1402] To a stirred solution of tert-butyl (tert-butoxycarbonyl)(4-((4-((tert-butoxycarbonyl)amino)-3,3-dimethylbutyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 9 (70 mg, 0.096 mmol, 1.0 equiv) in CHCl (3 mL) was added HCl (4 M in dioxane, 5 mL) at 0°C and then stirred at room temperature for 2 h. The reaction progress was monitored by LCMS. The reaction mixture was concentrated under vacuum at below 40°C to give a light yellow solid. The solid obtained was dissolved in water (10 mL) and washed with MTBE (2×10 mL), the aqueous layer was lyophilized to afford 3-(4-((4-amino-3,3-dimethylbutyl)(4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl 124 (40 mg, 0.077 mmol, 80% yield) as an off-white solid.

[1403] 1 H-NMR (400MHz, DMSO-d6): δ11.00(s,1H),7.93(s,6H),7.34-7.44(m,1H),7.16-7.28(m,2H),5.11-5.16(m,1H),4.30-4.45(m, 2H),3.23(m,4H),2.90-2.98(m,1H),2.76-2.77(m,2H),2.64-2.68(m,3H),2.02-2.05(m,1H),1.52(m,7H),0.95-0.96(m,6H),

[1404] LCMS: 430.2 (M+H). Method: Column: Atlantis dC18 (50×4.6) mm, 5 μm, Mobile phase: A: 0.1% FA in H 2 O, Mobile phase: B: acetonitrile, Flow rate: 1.5 mL / min.

[1405] HPLC: Method: Mobile phase: 0.1% TFA in water, Mobile phase B: acetonitrile, Flow rate: 2.0 mL / min. Column: X-Bridge C8 (50×4.6) mm, 3.5 μm. RT: 1.407 min, Area: 96.954%.

[1406] Compound 125 in the form of a hydrochloride salt

[1407] 3-(4-((5-amino-3,3-dimethylpentyl)(4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1408]

[1409] Step 1: 5-Ethoxy-3,3-dimethyl-5-oxopentanoic acid

[1410]

[1411] To an ice-cold solution of 4,4-dimethyldihydro-2H-pyran-2,6(3H)-dione 1 (10 g, 70.3 mmol, 1 eq) in ethanol (50 mL, 5 volumes) was added sodium ethoxide (0.479 g, 7.03 mmol, 0.1 eq) dropwise and stirred at 85 ° C for 16 h. After the reaction was complete (confirmed by LCMS analysis), the reaction mixture was distilled under reduced pressure. The resulting residue was poured into an ice-cold solution of saturated NaHCO solution (200 mL) and washed with EtOAc (2×50 mL). The separated aqueous layer was further acidified with aqueous HCl (6 N, 40 mL, pH~2) and extracted with EtOAc (2×50 mL). The combined organic extracts were washed with brine (200 mL), dried over Na 2 SO 4 , filtered and concentrated to afford 5-ethoxy-3,3-dimethyl-5-oxopentanoic acid 2 (6 g, 31.9 mmol, 45.3% yield) as a colorless liquid.

[1412] 1 H-NMR (400MHz, DMSO-d6): δ12.03(s,1H),4.04(q,J=7.20Hz,2H),2.37(s,2H),2.29(d,J=7.20Hz,2H),1.18(t,J=7.20Hz,3H),1.05(s,6H).

[1413] Step 2: Ethyl 5-amino-3,3-dimethyl-5-oxopentanoate

[1414]

[1415] To an ice-cold solution of 5-ethoxy-3,3-dimethyl-5-oxopentanoic acid 2 (6 g, 31.9 mmol, 1 eq) in DCM (60 mL, 10 vol) was added thionyl chloride (15 mL, 206 mmol, 6 eq) dropwise followed by a catalytic amount of DMF (2 drops) and stirred at room temperature for 3.5 hours. The reaction was complete (by TLC analysis (30% EtOAc in petroleum ether, R f After the reaction mixture was distilled and dried under reduced pressure, the methyl ester was confirmed to be methyl ester.

[1416] The obtained acyl chloride mixture was dissolved in THF (30 mL) and added dropwise to a pre-cooled ammonia solution (0.5 M in THF, 90 mL, 4159 mmol) at -78 ° C. The reaction mixture was warmed to room temperature and stirred for 30 minutes. The reaction was completed (by TLC analysis (30% EtOAc in petroleum ether, R fAfter 3 hours of drying (10 minutes after 4 hours of drying), the reaction mixture was concentrated and the resulting residue was poured into an ice-cold solution of saturated NaHCO solution (150 mL) and extracted with EtOAc (2×50 mL). The combined organic layers were washed with brine (150 mL), dried over NaSO, filtered and concentrated to afford ethyl 5-amino-3,3-dimethyl-5-oxopentanoate 3 (4.4 g, 159 mmol, 50% yield) as a yellow liquid.

[1417] 1 H-NMR (400MHz, DMSO-d6): δ7.21(s,1H),6.73(s,1H),4.04(q,J=7.20Hz,2H),2.36(s,2H),2.08(s,2H),1.18(t,J=7.20Hz,3H),1.03(s,6H).

[1418] LCMS: 188.1 (M+H), Rt (min): 1.694, Area %: 99.678.

[1419] Step 3: 5-Amino-3,3-dimethylpentan-1-ol

[1420]

[1421] A solution of ethyl 5-amino-3,3-dimethyl-5-oxopentanoate 3 (4.4 g, 23.50 mmol, 1 eq) in THF (15 mL, 8 vol) was treated dropwise with LAH (2 M in THF, 23.50 mL, 47.0 mmol, 2 eq) at 0 °C over a period of 5 minutes. Once the addition was complete, the reaction mixture was allowed to warm to room temperature and then slowly heated to 65 °C and stirred for 2 hours. The reaction was complete (by TLC analysis (EtOAc, R f After 4 hours, the reaction mixture was cooled to 0 ° C and quenched slowly with saturated Na2SO4 solution (60 mL), the precipitated salts were removed by filtration through a celite pad and washed with EtOAc (50 mL). The combined filtrate was dried over Na2SO4, filtered and concentrated to obtain 5-amino-3,3-dimethylpentan-1-ol 4 (2 g, 13.30 mmol, 56.6% yield) as a brown liquid.

[1422] 1 H-NMR (400MHz, CDCl3): δ3.70(t,J=7.60Hz,3H),2.73(q,J=8.00Hz,2H),1.54(t,J=7.60Hz,2H),1.44-1.40(m,2H),1.02(s,1H),0.93(s,9H).

[1423] LCMS: 132.2 (M+H), Rt (min): 0.603, Area %: 87.229.

[1424] Step 4: tert-Butyl (5-hydroxy-3,3-dimethylpentyl)carbamate

[1425]

[1426] At room temperature, to 5-amino-3,3-dimethylpentan-1-ol 4 (2g, 15.24mmol, 1 equivalent) and sodium bicarbonate (2.56g, 30.5mmol, 2 equivalents) in a mixture of THF (30mL, 15 volumes) and water (2mL, 1 volume) stirred suspension dropwise add Boc-anhydride (3.54mL, 15.24mmol, 1 equivalent). Gained reaction mixture is stirred at room temperature for 16h. After reaction is completed (confirmed by LCMS analysis), anhydrous Na is added to the reaction mixture sO , filter and wash with EtOAc (20mL). Gained filtrate is concentrated under reduced pressure, to provide the crude product as a yellow liquid. The crude product was purified by column chromatography (Biotage R snap cartridge, KP-Sil, 100 g, 100-200 silica gel) using 20-25% EtOAc in petroleum ether to afford tert-butyl (5-hydroxy-3,3-dimethylpentyl)carbamate 5 (1.7 g, 7.33 mmol, 48.1% yield) as a yellow liquid.

[1427] 1 H-NMR (400MHz, DMSO-d6): δ6.70 (s, 1H), 3.43 (d, J = 4.40Hz, 2H), 2.94-2.87 (m, 2H), 1.44-1.20 (m, 16H), 0.78 (s, 6H).

[1428] LCMS: 132.3 (M-Boc), Rt (min): 2.14, Area %: 99.812.

[1429] Step 5: tert-Butyl (5-((tert-butyldimethylsilyl)oxy)-3,3-dimethylpentyl)carbamate

[1430]

[1431] To a solution of tert-butyl (5-hydroxy-3,3-dimethylpentyl)carbamate 5 (1.7 g, 7.35 mmol, 1.0 eq) and imidazole (1.251 g, 18.37 mmol, 2.5 eq) in DCM (20 mL) was added TBS-Cl (1.329 g, 8.82 mmol, 1.2 eq) at 0°C and allowed to stir at room temperature for 13 h. The reaction was complete (by TLC (30% EtOAc in petroleum ether, R f After 4 hours, 1 hour, 1 hour, 2 hours, 3 hours of drying, 1 hour after 4 ...

[1432] 1 H-NMR (400MHz, DMSO-d6): δ6.69(t,J=5.20Hz,1H),3.63(t,J=7.60Hz,2H),2.90(q,J=5 .60Hz,2H),1.40(s,10H),1.37(s,2H),0.85(t,J=3.20Hz,14H),0.03(q,J=2.80Hz,6H).

[1433] LCMS: 246.2 (M-Boc), Rt (min): 3.581, Area %: 99.821.

[1434] Step 6: tert-Butyl (tert-butoxycarbonyl)(5-((tert-butyldimethylsilyl)oxy)-3,3-dimethylpentyl)carbamate

[1435]

[1436] At 0 ° C, to a solution of tert-butyl (5-((tert-butyldimethylsilyl)oxy)-3,3-dimethylpentyl)carbamate 6 (2.5g, 7.23mmol) in THF (25mL, 10 volumes) was added n-BuLi (2.5M in hexane, 3.47mL, 8.68mmol, 1.2 equivalents). The reaction mixture was stirred at 0 ° C for 15 minutes. At the same temperature, a solution of (Boc) O (1.316mL, 5.67mmol, 1.2 equivalents) in THF (10mL) was added to the reaction mixture. The resulting reaction mixture was warmed to room temperature and stirred for 1 hour. The reaction was completed (by TLC analysis (5% EtOAc in petroleum ether, R fAfter 4 hours, 1 hour, 1 hour, 2 hours, 3 hours of drying, 1 hour after 4 hours, 3 hours of drying (1 hour after 4 hours, 1 hour after 3 hours, 1 hour after 4 hours, 3 hours after 3 hours, 4 hours after 5 hours, 5 hours after 8 hours, 8 hours after 8 hours, 1 hour after 4 hours, 2 hours after 8 hours, 3 hours after 8 hours, 4 hours after 3 hours, 5 hours after 4 hours, 3 hours after 5 hours, 2 hours after 8 hours, 3 hours after 3 hours, 4 hours after 5 hours, 5 hours after 3 hours, 3 hours after 4 hours, 5 hours after 3 hours, 4 hours after 5 hours, 3 hours after 3 hours, 4 hours after 3 hours, 5 hours after 4 hours, 3 hours after 5 hours, 4 hours after 3 hours, 5 hours after 3 hours, 4 hours after 5 hours, 5 hours after 3 hours, 6 hours after 4 hours, 7 hours after 5 hours, 8 hours after 3 hours, 6 hours after 3 hours, 7 hours after 3 hours, 8 hours after 3 hours, 4 hours after 5 hours, 5 hours after 3 hours, 6 hours after 3 hours, 7 hours after 3 hours, 8 hours after 3 hours, 4 hours after 5 hours, 7 hours after 3 hours, 8 hours after 3 hours, 5 hours after 3 hours, 6 hours after 3 hours, 7 hours after 3 hours, 8 hours after 4 hours, 8 hours after 3 hours, 9 hours after 4 hours, 9 hours after 3 hours, 1 hour after 3 hours, 1 hour after 3 hours, 2 hours after 3 hours, 3 hours after 4 hours, 5 hours after

[1437] 1 H-NMR (400MHz, DMSO-d6): δ1.47 (s, 9H), 1.44-1.41 (m, 4H), 0.91 (s, 4H), 0.86 (t, J = 4.40Hz, 6H), 0.03 (d, J = 4.80Hz, 3H).

[1438] LCMS: 246.3 (M-200), Rt (min): 4.602, Area %: 98.718.

[1439] Step 7: tert-Butyl (tert-butoxycarbonyl)(5-hydroxy-3,3-dimethylpentyl)carbamate

[1440]

[1441] To a solution of tert-butyl (tert-butoxycarbonyl)(5-((tert-butyldimethylsilyl)oxy)-3,3-dimethylpentyl)carbamate 7 (3.5 g, 7.85 mmol) in THF (35 mL, 10 vol) was added TBAF (11.78 mL, 1 M in THF, 11.78 mmol, 1.5 eq) and stirred at room temperature for 16 h. The reaction was complete (by TLC analysis (30% EtOAc in petroleum ether, R f To the 4-nitro-2-oxo-1-oxo-2-nitro-4-oxo-6 ...

[1442] 1H-NMR (400MHz, DMSO-d6): δ4.26 (t, J = 4.80Hz, 1H), 3.50-3.44 (m, 4H), 1.45 (s, 16H), 1.38 (q, J = 7.60Hz, 5H), 0.88 (s, 6H).

[1443] Step 8: tert-Butyl (tert-butoxycarbonyl)(3,3-dimethyl-5-oxopentyl)carbamate

[1444]

[1445] Under nitrogen atmosphere, at -78 DEG C, a solution of oxalyl chloride (0.317mL, 3.62mmol, 1.2 equivalents) in DCM (10mL) was added dropwise to a solution of DMSO (0.514mL, 7.24mmol, 2.4 equivalents), and stirred at the same temperature for 15 minutes. Over 5 minutes, a solution of (tert-butoxycarbonyl) (5-hydroxy-3,3-dimethylpentyl) tert-butyl carbamate 8 (1g, 3.02mmol, 1 equivalent) in DCM (10mL) was added dropwise thereto. After the reaction mixture was stirred at -78 DEG C for 30 minutes, TEA (2.103mL, 15.09mmol, 5 equivalents) was added dropwise at -78 DEG C. Once the addition was complete, the reaction mixture was slowly warmed to 0 DEG C. The reaction was completed (by TLC analysis (30% EtOAc in petroleum ether, R f ~0.8, KMnO4) confirmed), the reaction mixture was diluted with DCM (10 mL) and washed with 10% aqueous citric acid (50 mL), water (50 mL) and brine (50 mL), dried over Na2SO4, filtered and concentrated under vacuum below 30 ° C to give tert-butyl (tert-butoxycarbonyl)(3,3-dimethyl-5-oxopentyl)carbamate 9 (930 mg, 2.82 mmol, 94% yield) as a yellow liquid, which was used further without purification.

[1446] 1 H-NMR (400MHz, DMSO-d6): δ9.76 (q, J=2.40Hz, 1H), 3.49-3.44 (m, 2H), 2.29 (d, J=2.80 Hz,1H),1.52(d,J=8.00Hz,1H),1.49(s,12H),1.44(s,1H),1.02(s,3H),0.88(s,1H).

[1447] Step 9: tert-Butyl (tert-butoxycarbonyl)(5-((4-((tert-butoxycarbonyl)amino)butyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-3,3-dimethylpentyl)carbamate

[1448]

[1449] To a solution of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate Int-4 (1 g, 2.323 mmol), tert-butyl (tert-butoxycarbonyl)(3,3-dimethyl-5-oxopentyl)carbamate 9 (930 mg, 2.82 mmol, 1.2 eq) and TFA (0.716 mL, 9.29 mmol, 4 eq) in a mixture of dichloromethane (10 mL, 10 vol) and DMF (10 mL, 10 vol) was added sodium triacetoxyborohydride (1.969 g, 9.29 mmol, 4 eq) and stirred at room temperature for 16 h. TLC analysis (EtOAc, R f 0.094mmol, 4.04% yield).

[1450] 1 H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.38(t,J=7.60Hz,1H),7.19(d,J=7.20Hz,1H), 7.08(d,J=7.60Hz,1H),6.77(s,1H),5.12(q,J=5.20Hz,1H),4.40(d,J=16.80Hz,1H),4.29 (d,J=17.20Hz,1H),3.46(t,J=9.20Hz,2H),3.25(d,J=28.00Hz,4H),2.93(t,J=7.20Hz,3H ), 2.04 (t, J = 7.60Hz, 1H), 1.44 (t, J = 8.40Hz, 20H), 1.37 (d, J = 14.00Hz, 12H), 1.02 (s, 6H).

[1451] LC-MS: 744.5 (M+H), Rt (min): 3.010, Area %: 99.774.

[1452] Step 10: 3-(4-((5-amino-3,3-dimethylpentyl)(4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1453]

[1454] To a stirred suspension of tert-butyl (tert-butoxycarbonyl)(5-((4-((tert-butoxycarbonyl)amino)butyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-3,3-dimethylpentyl)carbamate 10 (70 mg, 0.094 mmol) in dichloromethane (3 mL) at 0°C was added HCl (4 M in dioxane, 1 mL, 4.00 mmol) and then stirred at room temperature for 2 h. The reaction was complete (by TLC analysis (EtOAc, R f After 10 min (confirmation of 4% HCl, 0.1% HCl), the reaction mixture was concentrated under reduced pressure. The obtained solid was dissolved in water (30 mL) and washed with MTBE (2×30 mL). The aqueous layer was lyophilized to obtain 3-(4-((5-amino-3,3-dimethylpentyl)(4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl 125 (12 mg, 0.023 mmol, 24.65% yield) as an off-white solid.

[1455] 1 H-NMR (400MHz, DMSO-d6): δ11.01(s,1H),8.00(s,6H),7.54(s,2H),5.17-5.12(m,1H),4.49(d,J=31.20Hz,2H),3.35(s,4H) ,2.92(d,J=12.40Hz,1H),2.75-2.61(m,5H),2.07(t,J=5.20Hz,1H),1.51(q,J=8.40Hz,6H),1.39-1.30(m,2H),0.87(s,6H).

[1456] LCMS: 444.3 (M+H). Method: Mobile phase A: 0.1% formic acid in H2O. Mobile phase B: ACN. Flow rate: 1.5 mL / min. Column: Atlantis dC18 (50 x 4.6) 5 μm. Rt (min): 1.035; Area %: 99.826.

[1457] HPLC: Method: Mobile phase A: 0.1% formic acid in H2O. Mobile phase B: methanol. Flow rate: 1.0 mL / min. Column: X-Bridge C8 (50 × 4.6 mm, 3.5 μm). Rt (min): 2.679; Area %: 98.409.

[1458] Compound 126 in the form of a hydrochloride salt

[1459] 3-(4-((5-amino-4,4-dimethylpentyl)(4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1460]

[1461] Step 1: 5-((tert-Butyldimethylsilyl)oxy)-2,2-dimethylvaleronitrile

[1462]

[1463] A solution of LDA (2M in THF / heptane / ethylbenzene, 5.92 mL, 11.85 mmol, 1.5 equiv) in THF (10 mL) was cooled to -78 ° C and isobutyronitrile 2 (0.764 g, 11.06 mmol, 1.4 equiv) was added over 5 minutes. The reaction mixture was warmed to -40 ° C and stirred for 30 minutes and cooled to -78 ° C again. (3-bromopropoxy) (tert-butyl) dimethylsilane 1 (2.0 g, 7.90 mmol, 1.0 equiv) was then added dropwise to the reaction mixture and allowed to warm to 0 ° C and stirred for 1 h. The reaction was complete (by TLC and UPLC analysis (10% EtOAc / petroleum ether, R f After 4 hours, 5-(tert-butyldimethylsilyl)oxy)-2,2-dimethylvaleronitrile (2.2 g, 9.11 mmol, 115% yield) was obtained as a light yellow liquid, which was used further without purification.

[1464] 1 H-NMR (400MHz, DMSO-d6): δ3.66-3.69(m,2H),1.69-1.73(m,2H),1.59-1.63(m,2H),1.38(s,6H),0.91(s,9H),0.08(s,6H)

[1465] Step 2: tert-Butyl (5-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpentyl)carbamate

[1466]

[1467] To a stirred solution of 5-((tert-butyldimethylsilyl)oxy)-2,2-dimethylvaleronitrile 3 (1.0 g, 4.14 mmol, 1.0 equiv) in dry MeOH (30 mL) cooled to 0° C. was added (Boc) O (1.923 mL, 8.28 mmol, 2.0 equiv), NiCl 6H O (0.984 g, 4.14 mmol, 1.0 equiv), and then carefully added NaBH (1.097 g, 29.0 mmol, 7.0 equiv) in small portions. The reaction was exothermic and bubbling. The resulting reaction mixture was allowed to warm to room temperature and stirred overnight. The reaction was complete (by TLC analysis (5% EtOAc / petroleum ether, R f After 4 hours, the reaction mixture was diluted with DCM (20 mL) and filtered through a bed of celite and washed with DCM (20 mL). The filtrate was washed with saturated NaHCO solution (1×20 mL), the organic layer was dried over NaSO and concentrated under reduced pressure to give the crude product (tert-butyl 5-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpentyl)carbamate 4 (1.2 g, 3.39 mmol, 82% yield) as a light yellow liquid, which was used further without purification.

[1468] 1 H-NMR (400MHz, DMSO-d6): δ6.71(t,J=6.40Hz,1H),3.54(m,2H),2.73-2.76(m,2H),1.4 4(s,9H),1.22-1.38(m,2H),1.07-1.13(m,2H),0.89(s,9H),0.80(s,6H),0.02(s,6H).

[1469] Step 3: tert-Butyl (tert-butoxycarbonyl)(5-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpentyl)carbamate

[1470]

[1471] At 0 ° C, to a stirred solution of tert-butyl (5-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpentyl)carbamate 4 (2.5g, 7.23mmol, 1.0 equivalent) in THF (50mL) was added n-BuLi (2.5M in hexane, 3.47mL, 8.68mmol, 1.2 equivalents). The reaction mixture was stirred at 0 ° C for 15 minutes. At the same temperature, a solution of (Boc)2O (2.015mL, 8.68mmol, 1.2 equivalents) in THF (10mL) was added to the reaction mixture. The resulting reaction mixture was warmed to room temperature and stirred for 1h. The reaction was completed (by TLC analysis (10% EtOAc / petroleum ether, R f After 4 hours of drying (confirmed by adding 4% NaHCO₄) and 8% NaHCO₄), the reaction mixture was quenched with water (50 mL) and extracted with DCM (2×100 mL). The combined organic layers were dried over Na₂SO₄ and concentrated under reduced pressure to afford tert-butyl (tert-butoxycarbonyl)(5-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpentyl)carbamate 5 (3.2 g, 6.82 mmol, 94% yield) as a light yellow liquid, which was used further without purification.

[1472] 1 H-NMR (400MHz, DMSO-d6): δ3.59-3.54(m,2H),2.73-2.76(m,2H),1.42(s,18H) ,1.24-1.36(m,2H),1.07-1.13(m,2H),0.89(s,9H),0.80(s,6H),0.02(s,6H).

[1473] Step 4: tert-Butyl (tert-butoxycarbonyl)(5-hydroxy-2,2-dimethylpentyl)carbamate

[1474]

[1475] To a stirred solution of tert-butyl (tert-butoxycarbonyl)(5-((tert-butyldimethylsilyl)oxy)-2,2-dimethylpentyl)carbamate (3.2 g, 7.18 mmol, 1.0 eq) in THF (60 mL) was added TBAF (1 M in THF, 10.8 mL, 10.8 mmol, 1.5 eq) at room temperature, and the resulting solution was stirred at room temperature overnight. The reaction was complete (by TLC analysis (20% EtOAc / petroleum ether, R f4.82mmol, 67.1% yield)。

[1476] 1 H-NMR (400MHz, DMSO-d6): δ4.35 (t, J=5.20Hz, 1H), 3.36-3.39 (m, 2H), 3.32-3 .35(m,2H),1.47(s,18H),1.36-1.40(m,2H),1.12-1.16(m,2H),0.79(s,6H).

[1477] Step 5: tert-Butyl (tert-butoxycarbonyl)(2,2-dimethyl-5-oxopentyl)carbamate

[1478]

[1479] At -78 ℃, to oxalyl chloride (0.845mL, 9.65mmol, 2.0 equivalents) and CH cl (20mL) solution adds DMSO (1.370mL, 19.31mmol, 4.0 equivalents) CH cl (5mL) solution, and stirred 15 minutes at the same temperature. At -78 ℃, to this mixture, add (tert-butoxycarbonyl) (5-hydroxy-2,2-dimethylpentyl) t-butyl carbamate 6 (1.6g, 4.83mmol, 1.0 equivalents) in CH cl (5mL) solution in. After stirring reaction mixture 30 minutes at -78 ℃, at -78 ℃, dropwise add triethylamine (4.04mL, 29.0mmol, 6.0 equivalents). Then, reaction mixture is slowly warmed to 0 ℃ and stirred 30 minutes. Reaction is completed (by TLC analysis (20% EtOAc / petroleum ether, R f~0.6, KMnO4) confirmed), the reaction mixture was diluted with DCM (40 mL) and washed with 10% aqueous citric acid (1×40 mL), water (1×40 mL), dried over Na2SO4, and concentrated under vacuum (below 30°C) to afford tert-butyl (tert-butoxycarbonyl)(2,2-dimethyl-5-oxopentyl)carbamate 7 (1.55 g, 3.42 mmol, 70.8% yield) as a light yellow liquid, which was used further without purification.

[1480] 1 H-NMR (400MHz, DMSO-d6): δ9.68(s,1H),3.39(s,2H),2.42-2.50(m,2H),1.44(m,2H),1.42(s,18H),0.79(s,6H)

[1481] Step 6: tert-Butyl (tert-butoxycarbonyl)(5-((4-((tert-butoxycarbonyl)amino)butyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-2,2-dimethylpentyl)carbamate

[1482]

[1483] To a stirred mixture of tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)butyl)carbamate 8 (0.5 g, 1.161 mmol, 1.0 eq) and tert-butyl (tert-butoxycarbonyl)(2,2-dimethyl-5-oxopentyl)carbamate 7 (0.765 g, 2.323 mmol, 2.0 eq) in CHCl (10 mL) and DMF (10 mL) was added trifluoroacetic acid (0.358 mL, 4.65 mmol, 4.0 eq) and stirred at 0°C for 10 minutes. To this mixture was added sodium triacetoxyborohydride (0.985 g, 4.65 mmol, 4.0 eq) in portions at 0°C, and the resulting colorless turbid material was stirred at room temperature for 16 h. The reaction was complete (by TLC analysis (100% EtOAc, R f7-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-2,2-dimethylpentyl)carbamic acid tert-butyl ester 9 (0.25g, 0.335mmol, 28.9% yield) was obtained.

[1484] 1 H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.35-7.39(m,1H),7.06-7.19(m,2H),6.77-6.79(m,1H),5.09-5.13(m,1H),4.27-4.42( m,2H),3.10-3.21(m,4H),2.89-2.90(m,4H),2.67-2.68(m,6H),1.44(s,18H),1.43(m,4H),1.35(s,9H),1.24(m,2H),0.75(s,6H),

[1485] LCMS: 744.5 (M+H). Method: Atlantis dC18 (50×4.6 mm) 5 μm, mobile phase: A: 0.1% FA in H2O, mobile phase: B: acetonitrile, flow rate: 1.5 mL / min

[1486] HPLC: Method: Mobile phase A: 0.1% FA in water, Mobile phase B: acetonitrile, Flow rate: 2.0 mL / min. Column: X-Bridge C8 (50×4.6) mm, 3.5 μm. RT: 6.191 min, Area: 96.037%

[1487] Step 7: 3-(4-((5-amino-4,4-dimethylpentyl)(4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione dihydrochloride

[1488]

[1489] To a stirred solution of tert-butyl (tert-butoxycarbonyl)(5-((4-((tert-butoxycarbonyl)amino)butyl)(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-2,2-dimethylpentyl)carbamate 9 (0.15 g, 0.202 mmol, 1.0 equiv) in dichloromethane (10 mL) was added HCl (4 M in dioxane, 10 mL) at 0°C and then stirred at room temperature for 2 h. The reaction progress was monitored by LCMS. The reaction mixture was concentrated under vacuum at below 40°C to give a light yellow solid. The obtained solid was dissolved in water (10 mL) and washed with MTBE (1×10 mL), the aqueous layer was lyophilized to afford 3-(4-((5-amino-4,4-dimethylpentyl)(4-aminobutyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 2HCl (80 mg, 0.148 mmol, 73.4% yield) as a light yellow solid.

[1490] 1 H-NMR (400MHz, DMSO-d6): δ10.99(s,1H),7.94(s,6H),7.43(m,1H),7.25-7.28(m,2H),5.11-5.15(m,1H),4.12-4.50(m,2H),3.20-3.26(m, 4H),2.90-2.98(m,1H),2.76-2.89(m,2H),2.64-2.68(m,3H),2.02-2 .05(m,1H),1.54(m,4H),1.39(m,2H),1.24-1.26(m,3H),0.93(s,6H),

[1491] LCMS: 444.4 (M+H). Method: Column: Atlantis dC18 (50×4.6 mm) 5 μm, Mobile phase: A: 0.1% FA in H 2 O, Mobile phase: B: acetonitrile, Flow rate: 1.5 mL / min.

[1492] HPLC: Method: Mobile phase A: 0.1% TFA in water, Mobile phase B: Methanol, Flow rate: 1.0 mL / min. Column: X-Bridge C8 (50×4.6) mm, 3.5 μm. RT: 3.466 min, Area: 95.684%.

[1493] Compound 143

[1494] 1-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)-3-methylurea

[1495]

[1496] To a solution of 3-(4-((4-aminobutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl (200 mg, 0.499 mmol, 1 eq) and N-methyl-1H-imidazole-1-carboxamide (62.5 mg, 0.499 mmol, 1 eq) in DCM (4 mL) was added TEA (0.077 mL, 0.549 mmol, 1.1 eq) and the resulting reaction mixture was stirred at room temperature for 12 h. After completion of the reaction (confirmed by UPLC), the reaction mixture was concentrated under vacuum to obtain the crude product, which was purified by preparative HPLC (column: XBridge C8-150; method: HCl and ACN, flow rate: 20 mL / min) to afford 1-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)-3-methylurea (45 mg, 0.098 mmol, 19.56% yield) as a white solid.

[1497] 1 H-NMR (400MHz, DMSO-d6): δ11.03(s,1H),7.61(s,2H),5.13(q,J=4.80Hz,1H),4.74(s,2H),2.96-2.88(m,3H),2.65(q,J=17.60 Hz, 1H), 2.43 (q, J = 4.00Hz, 1H), 2.08 (t, J = 5.60Hz, 1H), 1.37 (t, J = 6.00Hz, 5H), 1.22 (d, J = 2.80Hz, 5H), 0.81 (t, J = 6.80Hz, 3H).

[1498] LCMS: 458.9 (M+H), method: mobile phase: A: 0.1% formic acid in H2O solution B: ACN; column: Atlantis dC18 (50×4.6) 5μm; flow rate: 1.5mL / min; Rt (min): 1.830, area%: 99.292.

[1499] HPLC: Method: Mobile phase: A: 0.1% FA in water, mobile phase D: ACN; Column: XBridge C8 (50×4.6) mm, 3.5 μm, Flow rate: 2.0 mL / min; Rt (min): 2.870, Area %: 99.180.

[1500] Compound 148

[1501] N-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)butanamide

[1502]

[1503] To a stirred solution of butyric acid 148a (0.150 g, 0.375 mmol, 1 eq) in DMF (2 mL) at 0° C., DIPEA (0.262 mL, 1.498 mmol, 4 eq) and HATU (0.214 g, 0.562 mmol, 1.5 eq) were added. 3-(4-((4-aminobutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl 47 (0.150 g, 0.375 mmol, 1 eq) was then added at the same temperature, and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was purified by preparative HPLC (instrument: SC-DC-ARD-05-044; flow rate: 15 mL / min; column: X-Select-C18-19 x 150 mm; mobile phase combination: 0.1% FA in water / MeOH). Pure fractions were lyophilized to afford N-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)butanamide 148 (37.43 mg, 0.079 mmol, 20.99% yield) as an off-white solid.

[1504] LCMS: 471.2 (M+H). Method: Column: Atlantis dC18 (50×4.6) 5 μm, Mobile Phase: A: 0.1% formic acid in H2O, B: ACN, Flow Rate: 1.5 mL / min. Rt (min): 2.127; Area % -99.68.

[1505] HPLC: 98.85%, Rt (min): 3.792. Method: Column: X-Bridge C8 (50×4.6) mm, 3.5 μm, Mobile phase: A: 0.1% FA in water, Mobile phase: B: ACN, Flow rate: 2.0 mL / min.

[1506] 1H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.71(d,J=5.20Hz,1H),7.37(t,J=7.60Hz,1H),7.19 (d,J=7.20Hz,1H),7.07(d,J=8.00Hz,1H),5.10(q,J=4.80Hz,1H),4.34(dd,J=16.80,43.20Hz, 2H),3.17(t,J=7.20Hz,4H),3.02(d,J=6.00Hz,2H),2.91(t,J=12.80Hz,1H),2.68(s,1H),2.6 2(s,1H),1.99(t,J=7.20Hz,3H),1.48-1.41(m,8H),1.25(d,J=3.20Hz,4H),0.86-0.80(m,6H).

[1507] Compound 149

[1508] N-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)-3-methylbutanamide

[1509]

[1510] To a stirred solution of 3-methylbutanoic acid 149a (35.1 mg, 0.343 mmol, 1 eq) in DMF (3 mL) at 0° C. was added HATU (196 mg, 0.515 mmol, 1.5 eq) and DIPEA (177 mg, 1.373 mmol, 4 eq), followed by 3-(4-((4-aminobutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl 47 (150 mg, 0.343 mmol, 1 eq), and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by UPLC), the reaction mixture was purified by reverse phase column chromatography (Grace column: C18 40 μm, 40 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN aqueous solution mobile phase). Pure fractions were lyophilized to afford N-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)-3-methylbutanamide 149 (51.28 mg, 0.106 mmol, 30.8% yield) as an off-white solid.

[1511] 1H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.73(d,J=5.20Hz,1H),7.37(t,J=7.60Hz,1H),7.18(d, J=7.20Hz,1H),7.07(d,J=7.60Hz,1H),5.11(q,J=5.20Hz,1H),4.39(d,J=16.80Hz,1H),4.28(d,J= 16.80Hz,1H),3.16(t,J=6.80Hz,4H),3.02(d,J=5.60Hz,2H),2.92(s,1H),2.62(s,1H),2.01(t,J= 5.60Hz, 1H), 1.92 (t, J = 11.20Hz, 3H), 1.42 (d, J = 7.20Hz, 6H), 1.25 (s, 4H), 0.84 (t, J = 6.00Hz, 9H).

[1512] LCMS: 485.3 (M+H), method: mobile phase: A: 0.1% FA in H2O; mobile phase: B: ACN; column: Atlantis dC18 (50×4.6 mm) 5 μm; column: Atlantis dC18 (50×4.6 mm) 5 μm; flow rate: 1.5 mL / min, Rt (min): 2.252, area%: 99.867.

[1513] HPLC: Method: Mobile phase A: 0.1% TFA in water; Mobile phase D: ACN; Column: XBridge C8 (50×4.6) mm, 3.5 μm; Flow rate: 2.0 mL / min, Rt (min): 3.324 min, Area %: 99.501.

[1514] Compound 150

[1515] N-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)-3,3-dimethylbutanamide

[1516]

[1517] To a stirred solution of 3,3-dimethylbutanoic acid 150a (39.9 mg, 0.343 mmol, 1 eq) in DMF (3 mL) at 0° C. was added HATU (196 mg, 0.515 mmol, 1.5 eq) and DIPEA (177 mg, 1.373 mmol, 4 eq), followed by 3-(4-((4-aminobutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl 47 (150 mg, 0.343 mmol, 1 eq), and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by UPLC), the reaction mixture was purified by reverse phase column chromatography (Grace column: C18 40 μm, 40 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN aqueous solution mobile phase). Pure fractions were lyophilized to afford N-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)-3,3-dimethylbutanamide 150 as an off-white solid (40.2 mg, 0.080 mmol, 23.28% yield).

[1518] 1 H-NMR (400MHz, DMSO-d6): δ10.98(s,1H),7.69(d,J=5.20Hz,1H),7.38(t,J=5.6 0Hz,1H),7.21(t,J=6.00Hz,1H),7.11(s,1H),5.14-5.08(m,1H),4.43-4.27(m, 2H),3.18(d,J=6.00Hz,4H),3.01(d,J=4.80Hz,3H),2.62(s,1H),2.02(d,J=6.0 0Hz, 1H), 1.91 (d, J = 5.60Hz, 2H), 0.92 (d, J = 5.20Hz, 9H), 0.84 (d, J = 5.60Hz, 3H).

[1519] LCMS: 499.3 (M+H), method: mobile phase: A: 0.1% FA in H2O; mobile phase: B: ACN; column: Atlantis dC18 (50×4.6 mm) 5 μm; Rt (min): 2.340 min, area %: 99.116.

[1520] HPLC: Method: Mobile phase A: 0.1% TFA in water; Mobile phase D: ACN; Column: XBridge C8 (50×4.6) mm, 3.5 μm; Rt (min): 3.570, Area %: 98.021.

[1521] Compound 151

[1522] 2-Cyclohexyl-N-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)acetamide

[1523]

[1524] To a stirred solution of 151a (0.150 g, 0.375 mmol, 1 eq) in DMF (2 mL) was added DIPEA (0.262 mL, 1.498 mmol, 4 eq) and HATU (0.214 g, 0.562 mmol, 1.5 eq) at 0°C. 3-(4-((4-aminobutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, HCl 47 (0.150 g, 0.375 mmol, 1 eq) was then added at the same temperature and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was purified by reverse phase column chromatography ( The purified product was purified by HPLC-MS / MS (column: C18 40 μm, 40 g; flow rate: 20 mL / min; 0.1% HCOOH / ACN in water mobile phase). Pure fractions were lyophilized to afford 2-cyclohexyl-N-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)(pentyl)amino)butyl)acetamide 151 (95.22 mg, 0.180 mmol, 48.2% yield) as an off-white solid.

[1525] LCMS: 525.3 (M+H). Method: Column: Atlantis dC18 (50×4.6) 5 μm, Mobile Phase: A: 0.1% TFA in H2O, B: 0.1% TFA in ACN, Flow Rate: 1.5 mL / min. Rt (min): 1.912; Area % -99.21.

[1526] HPLC: 99.40%, Rt (min): 4.020. Method: Column: X-Bridge C8 (50×4.6) mm, 3.5 μm, Mobile phase: A: 0.1% TFA in water, Mobile phase: B: ACN, Flow rate: 2.0 mL / min.

[1527] 1H-NMR (400MHz, DMSO-d6): δ10.99(s,1H),7.72(s,1H),7.37(t,J=7.60Hz,1H),7.18(d,J=7.20Hz,1H),7.0 6(d,J=8.40Hz,1H),5.11(q,J=4.80Hz,1H),4.39(d,J=16.80Hz,1H),4.28(d,J=16.80Hz,1H),3.17(d,J=6 .00Hz,4H),3.01(d,J=6.00Hz,2H),2.97-2.83(m,1H),2.01(t,J=6.00Hz,1H),1.89(d,J=6.80Hz,2H),1.5 9(d,J=8.80Hz,6H),1.42(d,J=6.80Hz,6H),1.25(s,4H),1.15(t,J=12.40Hz,3H),0.84(t,J=7.20Hz,5H).

[1528] The hydrochloride form of compound 163

[1529] 3-(4-((4-(Benzylamino)butyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione.HCl

[1530]

[1531] To a solution of 3-(4-((4-aminobutyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione.HCl 47 (200 mg, 0.500 mmol, 1 eq) and benzaldehyde (55.6 mg, 21.97 mmol, 1.05 eq) in DCM (4 mL) and DMF (1 mL) at 0°C was added TFA (0.034 mL, 0.430 mmol, 0.9 eq) and sodium triacetoxyborohydride (154 mg, 0.750 mmol, 1.5 eq) portionwise, and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (confirmed by UPLC), the reaction mixture was diluted with DCM (20 mL), washed with water (2×10 mL), brine (10 mL), dried over anhydrous Na 2 SO 4 , filtered and concentrated to provide the crude product, which was purified by preparative HPLC (column: XBridge C8-150; method: HCl and ACN, flow rate: 20 mL / min) to afford 3-(4-((4-(benzylamino)butyl)(pentyl)amino)-1-oxoisoindolin-2-yl)piperidine-2,6-dione 163 (29 mg, 0.058 mmol, 23.23% yield) as an off-white solid.

[1532] 1 H-NMR (400MHz, DMSO-d6): δ10.99 (s, 1H), 9.08 (s, 2H), 7.52 (q, J = 3.60Hz, 2H), 7.41-7.4 5(m,4H),7.26(s,2H),5.12(q,J=5.20Hz,1H),4.60-4.20(m,2H),4.09(t,J=6.00Hz,2H) ,3.22(s,2H),2.92-2.87(m,3H),2.63(t,J=18.00Hz,1H),2.33(d,J=1.60Hz,1H),1.64( t,J=7.60Hz,2H),1.45(d,J=24.80Hz,4H),1.28-1.21(m,4.8H),0.84(t,J=6.80Hz,3H).

[1533] LCMS: 491.2 (M+H), method: mobile phase: A: 0.1% TFA in H2O; mobile phase: B: 0.1% TFA in ACN; column: XBridge C8 (50×4.6 mm) 3.5 μm; flow rate: 1.5 mL / min; Rt (min): 1.852, area %: 98.144.

[1534] HPLC: Method: Mobile phase A: 0.1% TFA in water, Mobile phase D: ACN...

Claims

1. A compound or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein the compound is of formula (I): in, R 1 is: C1-C8 alkyl, C3-C8 alkenyl, C3-C8 alkynyl, C3-C8 cycloalkyl or C5-C 10 bicycloalkyl, each optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, NH2, C1-C3 haloalkyl, optionally substituted C6-C 10 Aryl, C1-C3 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C5-C 10 Bicycloalkyl or optionally substituted C3-C6 heterocycloalkyl; X is: a bond; or –(CH2) n –, optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, C1-C3 haloalkyl, C3-C6 cycloalkyl or C1-C6 alkyl, wherein the C1-C6 alkyl is able to form a C3-C6 spiroalkyl ring together with the atom to which it is attached, and wherein, –(CH2) n – The group can contain 0-1 double or triple bonds; n is an integer from 1 to 6; A is: bond; C4-C8 cycloalkyl, C5-C 10 Bicycloalkyl, C5-C 10 Heterobicycloalkyl or C3-C8heterocycloalkyl, each optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, CN, C1-C3 haloalkyl, C1-C3 alkyl or C3-C5 cycloalkyl; Y is a bond or –(CH2) m – m is an integer of 1 or 2; Z is H or halogen; R 2 Yes:-NR 12 R 5 、-NHC(O)R 6 、-NHC(O)NR 8 (R 9 )、-C(O)NHR 5 , a five-membered or six-membered heteroaryl or a C3-C8 heterocycloalkyl ring or a C5-C 10 a heterobicycloalkyl ring, optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, CN, C1-C3 haloalkyl, C1-C3 alkyl or C3-C5 cycloalkyl; R 5 is: H, C1-C5 alkyl, C3-C7 cycloalkyl, five-membered or six-membered heteroaryl, C5-C 10 Bicycloalkyl, (C3-C6 cycloalkyl)-C1-C3 alkyl, (C3-C6 heterocycloalkyl)-C1-C3 alkyl, (C6-C 10 (C1-C5-heteroaryl)-C1-C3 alkyl, or C3-C7 heterocycloalkyl, each optionally substituted with one or more halogen, CN, OH, O-(C1-C3 haloalkyl), O-(C1-C3 alkyl), C1-C3 haloalkyl, CH3SO2-, optionally substituted C3-C6 cycloalkyl, optionally substituted C3-C7 heterocycloalkyl, or C(O)NR 8 (R 9 ); R 6 is: C1-C6 alkyl, C3-C6 cycloalkyl or C3-C6 heterocycloalkyl, each optionally substituted with one or more halogen, C1-C3 haloalkyl, optionally substituted C3-C6 cycloalkyl or NR 8 (R 9 ); R 8 and R 9 are all independently H, substituted C1-C3 alkyl, or together with the nitrogen to which they are attached form a 4-6 membered heterocyclic ring optionally substituted with one or more halogen or C1-C3 haloalkyl; R 12 is: H, C1-C5 alkyl, or the nitrogen to which it is attached and R 5 Together they form (i) a C3-C8 cycloalkyl ring or a heterocycloalkyl ring or (ii) a C5-C 10 A bicycloalkyl ring or a heterobicycloalkyl ring, each optionally substituted with one or more halogen, C1-C3 haloalkyl, OH, O-(C1-C3 alkyl), O-(C1-C3 haloalkyl) or CN, Each stereocenter in the compound of formula (I) is independently an R-enantiomer, an S-enantiomer, or a mixture of an R-enantiomer and an S-enantiomer, and Each double bond in the compounds of formula (I) is independently cis or trans.

2. The compound according to claim 1 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: The compound is represented by formula (Ia), and X is a bond or –(CH2) n –, 3. The compound according to claim 2 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: R 1 It is: C1-C8 alkyl or C3-C8 cycloalkyl, each optionally substituted with one or more halogen, OH, NH2, C1-C3 alkyl, C1-C3 haloalkyl, O-(C1-C3 alkyl), optionally substituted C6 aryl, optionally substituted C3-C8 cycloalkyl, optionally substituted C3-C6 heterocycloalkyl or optionally substituted C6-C8 bicycloalkyl.

4. The compound according to claim 2 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: A is: C4-C6 cycloalkyl, C4-C6 heterocycloalkyl or C5-C7 bicycloalkyl, each optionally substituted with one or more halogen, OH, C1-C3 alkyl or C1-C3 haloalkyl.

5. The compound according to claim 2 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: Y is a bond or -CH2-.

6. The compound according to claim 2 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: R 5 is: H, C1-C5 alkyl, C3-C6 cycloalkyl, 6-membered heteroaryl, C5 bicycloalkyl, (C3-C4 cycloalkyl)-C1-C2 alkyl, (C3-C4 heterocycloalkyl)-C1-C2 alkyl, (C6 aryl)-C1-C2 alkyl, (C5 heteroaryl)-C1-C2 alkyl or C3-C4 heterocycloalkyl, each optionally substituted with one or more halogen, CN, OH, O-(C1-C3 haloalkyl), O-(C1-C3 alkyl), C1-C3 haloalkyl, CH3SO2- or C(O)NR 8 (R 9 ).

7. The compound according to claim 2 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: R 12 is H, C1-C5 alkyl, or the nitrogen to which it is attached and R 5 Together they form a C3-C8 heterocycloalkyl ring optionally substituted with one or more halogen or C1-C3 haloalkyl.

8. The compound according to claim 1 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: The compound is represented by formula (Ib), and X is a bond or –(CH2) n –, 9. The compound according to claim 8 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: R 1 is: C3-C6 alkyl, optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, NH2, C1-C3 haloalkyl.

10. The compound according to claim 8 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: A is: a bond; or C4-C6 cycloalkyl, optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, NH2, C1-C3 haloalkyl.

11. The compound according to claim 8 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: Y is a bond.

12. The compound according to claim 8 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: R 6 is optionally replaced with NR 8 (R 9 )'s C1-C6 alkyl.

13. The compound according to claim 1 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: The compound is represented by formula (Ic), and X is a bond or –(CH2) n –, 14. The compound according to claim 13 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: R 1 is: C1-C6 alkyl, optionally substituted with optionally substituted C3-C8 cycloalkyl or optionally substituted C3-C6 heterocycloalkyl.

15. The compound according to claim 13 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: A is: a bond; C4-C6 cycloalkyl, C4-C6 heterocycloalkyl or C5-C7 bicycloalkyl, each optionally substituted with one or more halogen, OH, O-C1-C3 alkyl, NH2, C1-C3 haloalkyl.

16. The compound according to claim 13 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: Y is a bond.

17. The compound according to claim 13, or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: R 5 It is H or C1-C3 alkyl.

18. The compound according to claim 1 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: Each X and Y is a bond; A is a C4-C6 cycloalkyl group; R 1 is C1-C3 alkyl optionally substituted with optionally substituted C3-C6 cycloalkyl; and R 2 It is a 5-membered heteroaryl.

19. A compound or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein the compound is of formula (Id): in, R 1 is C1-C3 alkyl optionally substituted with optionally substituted C3-C6 cycloalkyl; A is an optionally substituted C4-C6 cycloalkyl; R 2 is: C1-C4 alkyl or (C3-C4 cycloalkyl)-C1-C2 alkyl, each optionally substituted with one or more halogen, CN, OH, O-(C1-C3 haloalkyl), O-(C1-C3 alkyl), C1-C3 haloalkyl or CH3SO2-; Each stereocenter in the compound of formula (Id) is independently the R-enantiomer, the S-enantiomer, or a mixture of the R-enantiomer and the S-enantiomer.

20. The compound according to claim 19, or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein: R 2 is C1-C3 alkyl optionally substituted with one or more halogen, O-(C1-C3 haloalkyl), O-(C1-C3 alkyl) or C1-C3 haloalkyl.

21. The compound according to claim 1 or 19, wherein The compounds are used to degrade or reduce casein kinase 1 alpha (CK1 alpha).

22. The compound according to claim 1 or 19, wherein The compounds are used to inhibit casein kinase 1α (CK1α) activity.

23. The compound according to claim 1 or 19, wherein The compounds are used for preventing or treating blood cancer.

24. A pharmaceutical composition, comprising: A compound according to claim 1 or 19, or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, and Optionally, a pharmaceutically acceptable excipient or carrier.

25. The pharmaceutical composition according to claim 24, wherein The pharmaceutical composition is used for degrading or reducing casein kinase 1α (CK1α).

26. The pharmaceutical composition according to claim 24, wherein The pharmaceutical composition is used for inhibiting the activity of casein kinase 1α (CK1α).

27. The pharmaceutical composition according to claim 24, wherein The pharmaceutical composition is used for preventing or treating blood cancer.

28. The pharmaceutical composition according to claim 27, wherein The blood cancer is leukemia, lymphoma or myeloma.

29. The pharmaceutical composition according to claim 28, wherein The leukemia is selected from the group consisting of acute myeloid leukemia (AML), acute lymphoblastic leukemia or acute lymphocytic leukemia (ALL), T-cell acute lymphocytic leukemia (T-ALL), B-cell acute lymphocytic leukemia (B-ALL), chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), T-cell prolymphocytic leukemia (T-PLL), large granular lymphocytic leukemia, adult T-cell leukemia, chronic eosinophilic leukemia (CEL) and myelodysplastic syndrome (MDS).

30. The pharmaceutical composition according to claim 28, wherein The lymphoma is selected from the group consisting of Hodgkin lymphoma, diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, small lymphocytic lymphoma (SLL), mantle cell lymphoma (MCL), marginal zone lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, primary central nervous system (CNS) lymphoma and peripheral T-cell lymphoma.

31. The pharmaceutical composition according to claim 28, wherein The myeloma is selected from the group consisting of multiple myeloma, light chain myeloma, non-secretory myeloma, solitary plasmacytoma, extramedullary plasmacytoma, monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM), immunoglobulin D (IgD) myeloma and immunoglobulin E (IgE) myeloma.

32. A method of degrading or reducing casein kinase 1 alpha (CK1 alpha) in a subject in need thereof, the method comprising administering to the subject an effective amount of the pharmaceutical composition of claim 24.

33. A method of inhibiting casein kinase 1 alpha (CK1 alpha) activity in a subject in need thereof, the method comprising administering to the subject an effective amount of the pharmaceutical composition of claim 24.

34. A method for preventing or treating blood cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of the pharmaceutical composition according to claim 24.

35. The method of claim 34, wherein: The blood cancer is leukemia, lymphoma or myeloma.

36. The method of claim 35, wherein: The leukemia is selected from the group consisting of acute myeloid leukemia (AML), acute lymphoblastic leukemia or acute lymphocytic leukemia (ALL), T-cell acute lymphocytic leukemia (T-ALL), B-cell acute lymphocytic leukemia (B-ALL), chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), T-cell prolymphocytic leukemia (T-PLL), large granular lymphocytic leukemia, adult T-cell leukemia, chronic eosinophilic leukemia (CEL) and myelodysplastic syndrome (MDS).

37. The method of claim 35, wherein: The lymphoma is selected from the group consisting of Hodgkin lymphoma, diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, small lymphocytic lymphoma (SLL), mantle cell lymphoma (MCL), marginal zone lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, primary central nervous system (CNS) lymphoma and peripheral T-cell lymphoma.

38. The method of claim 35, wherein the myeloma is selected from the group consisting of multiple myeloma, light chain myeloma, non-secretory myeloma, solitary plasmacytoma, extramedullary plasmacytoma, monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM), immunoglobulin D (IgD) myeloma, and immunoglobulin E (IgE) myeloma.

39. A compound or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof, wherein the compound is: [Table A] 40. A pharmaceutical composition, comprising: The compound according to claim 39 or a pharmaceutically acceptable salt, enantiomer, stereoisomer or prodrug thereof; as well as Optionally, a pharmaceutically acceptable excipient or carrier.

41. The pharmaceutical composition according to claim 40, wherein The pharmaceutical composition is used for degrading or reducing casein kinase 1α (CK1α).

42. The pharmaceutical composition according to claim 40, wherein The pharmaceutical composition is used for inhibiting the activity of casein kinase 1α (CK1α).

43. The pharmaceutical composition according to claim 40, wherein The pharmaceutical composition is used for preventing or treating blood cancer.

44. The pharmaceutical composition according to claim 40, wherein The blood cancer is leukemia, lymphoma or myeloma.

45. The pharmaceutical composition according to claim 44, wherein The leukemia is selected from the group consisting of acute myeloid leukemia (AML), acute lymphoblastic leukemia or acute lymphocytic leukemia (ALL), T-cell acute lymphocytic leukemia (T-ALL), B-cell acute lymphocytic leukemia (B-ALL), chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), T-cell prolymphocytic leukemia (T-PLL), large granular lymphocytic leukemia, adult T-cell leukemia, chronic eosinophilic leukemia (CEL) and myelodysplastic syndrome (MDS).

46. ​​The pharmaceutical composition according to claim 44, wherein The lymphoma is selected from the group consisting of Hodgkin lymphoma, diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, small lymphocytic lymphoma (SLL), mantle cell lymphoma (MCL), marginal zone lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, primary central nervous system (CNS) lymphoma and peripheral T-cell lymphoma.

47. The pharmaceutical composition according to claim 44, wherein The myeloma is selected from the group consisting of multiple myeloma, light chain myeloma, non-secretory myeloma, solitary plasmacytoma, extramedullary plasmacytoma, monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM), immunoglobulin D (IgD) myeloma and immunoglobulin E (IgE) myeloma.

48. A method of degrading or reducing casein kinase 1 alpha (CK1 alpha) in a subject in need thereof, the method comprising administering to the subject an effective amount of a composition comprising a compound according to claim 39.

49. A method of inhibiting casein kinase 1 alpha (CK1 alpha) activity in a subject in need thereof, the method comprising administering to the subject an effective amount of a composition comprising a compound according to claim 39.

50. A method of preventing or treating blood cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of a composition comprising a compound according to claim 39.

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