Fused tetracyclic compounds, compositions and uses thereof

Abstract

Provided herein are substituted fused tetracyclic compounds useful as inhibitors of MK2. The invention further provides pharmaceutical compositions of the compounds of the invention. The invention also provides medical uses of the substituted fused tetracyclic compounds.

Description

[Technical field]

[0001] Related Applications This application claims the benefit of priority to PCT International Application No. PCT / CN21 / 71062, filed January 11, 2021, which is hereby incorporated by reference in its entirety. [Background technology]

[0002] p38 mitogen-activated protein kinase (p38MAPK) transduces a range of extracellular signals that result in inflammatory responses, cell division and differentiation, apoptosis, and cell motility. p38MAPK was initially thought to be an ideal target for anti-inflammatory therapeutics. However, the failure of over a dozen chemically distinct compounds in the clinical phase suggests that p38MAPK may be an undesirable therapeutic target. Many of these compounds were found to be hepatotoxic to various degrees and resistant to anti-inflammatory effects that developed within weeks. Consequently, since p38MAPK controls activity towards over 60 substrates, failure in clinical trials due to undesirable side effects is perhaps not unexpected.

[0003] One of the downstream substrates of p38MAPK is mitogen-activated protein kinase-activated protein kinase-2 (MAPKAPK or MK2). Among other roles, MK2 controls the biosynthesis of tumor necrosis factor alpha and other cytokines. In addition, MK2 is activated after DNA damage, resulting in cell cycle arrest so that cells have the ability to repair DNA and continue to proliferate. MK2 also phosphorylates heat shock 27 (Hsp27), a prominent biomarker of cancer progression. Thus, MK2 serves as a promising anti-inflammatory and anti-cancer target and may improve the efficacy of chemotherapy without the unwanted side effects of affecting further upstream targets (i.e., p38MAPK). Summary of the Invention [Problem to be solved by the invention]

[0004] Thus, there is a continuing need to discover and develop new compounds that inhibit MK2 and may be useful therapeutics. [Means for solving the problem]

[0005] In certain embodiments, the present invention provides a compound having the structure of formula (I):

[0006] [ka] and pharma- ceutically acceptable salts thereof [wherein X 1 ~X 3 , Y 1 ~Y 3 and Z is as defined herein.

[0007] In some embodiments, the present invention relates to a pharmaceutical composition comprising a compound of formula (I) and a pharma- ceutically acceptable carrier.

[0008] The present invention also relates to a method of treating an MK2-associated disorder, comprising administering to a subject a compound of the present invention.

[0009] The present invention further relates to a method of inhibiting the proliferation of cancer cells, comprising contacting the cancer cells with a compound of the present invention.

[0010] The present invention also relates to a method of inhibiting MK2 activity in a cell, comprising contacting the cell with a compound of the present invention.

[0011] The present invention also relates to a method of treating or preventing a metabolic disorder, comprising administering to a subject a compound of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] In certain aspects, the present invention provides substituted fused tetracyclic compounds and pharmaceutical compositions thereof. In particular, such substituted fused tetracyclic compounds are useful as MK2 inhibitors and can therefore be used as anti-cancer, anti-inflammatory, or anti-diabetic agents.

[0013] I. Compound In certain embodiments, the present invention provides a compound having the structure of formula (I):

[0014] [ka] [In the formula, X 1 , X 2 , and X 3 are each independently CH, C-(alkyl), or N; Y 1 and Y 2 are each independently H or halogen; Y 3 is CN or a halogen; Z is -C(O)NZ 1 Z 2 , -NHC(O)NZ 3 Z 4 , -NHC(O)C(HZ 5 Z 6 ), or -NZ 7 Z 8 and; Z 1 is H or alkyl; Z 2 is an optionally substituted alkyl, aminoalkyl, cycloalkyl, heterocyclyl, or heterocycloalkyl; or Z 1 and Z 2 are joined together with the N to which they are attached to form an optionally substituted 4-, 5-, or 6-membered heterocycle or an optionally substituted heterobicycle; Z 3 and Z 4are joined together with the N to which they are attached to form an optionally substituted 4-, 5-, or 6-membered heterocycle; Z 5 and Z 6 are joined together with the C to which they are attached to form an optionally substituted 4-, 5-, or 6-membered heterocycle; Z 7 and Z 8 are linked together with the N to which they are attached to form an optionally substituted 5- or 6-membered cyclic amide, cyclic urea, or cyclic carbamate. Regarding.

[0015] In a particular embodiment, the present invention provides a method for producing

[0016] [ka] and pharma- ceutically acceptable salts thereof, wherein Z is as defined herein.

[0017] In certain embodiments, X 1 , X 2 and X 3 One of the is N, for example, X 1 is N and X 2 and X 3 Each is CH. In another embodiment, X 1 , X 2 and X 3 Two of them are N, for example, X 1 and X 3 are N and X 2 is CH or X 1 and X 2 are N and X 3 is CH or X 1 and X 2 are N and X 3 is CCH3. In another embodiment, X 1 , X 2 and X 3 Each of is N.

[0018] In certain embodiments, Z is -C(O)NZ 1 Z 2 It is.

[0019] In certain embodiments, Z 1 is H or CH3.

[0020] In certain embodiments, Z 2 is substituted alkyl or substituted aminoalkyl. In other embodiments, Z 2 is a substituted straight chain C1-C6 alkyl. 2 is a substituted branched C2-C6 alkyl.

[0021] In certain embodiments, Z 2 is substituted with alkylamino, e.g., methylamino or dimethylamino. In another embodiment, Z 2 is substituted with hydroxyalkyl.

[0022] In certain embodiments, Z is

[0023] [ka] and; n is 0, 1 or 2; R 1 , R 2 , R 3 , and R 4 is independently at each occurrence H, alkyl, or hydroxyalkyl; R 5 and R 6 is independently at each occurrence H or alkyl.

[0024] In certain embodiments, R 1 , R 2 , R 3 , and R 4 is independently at each occurrence H or CH; R 5 and R 6 Each occurrence of R is CH. 1, R 2 , R 3 , and R 4 is H for each occurrence; R 5 and R 6 is CH3 each time it occurs.

[0025] In certain embodiments, Z is

[0026] [ka] It is.

[0027] In certain embodiments, Z 2 is an optionally substituted heterocycloalkyl.

[0028] In certain embodiments, heterocycloalkyl includes azetidinyl, pyrrolidinyl, or piperidinyl. In other embodiments, azetidinyl, pyrrolidinyl, or piperidinyl, when substituted, are substituted with alkyl.

[0029] In certain embodiments, Z is

[0030] [ka] It is.

[0031] In certain embodiments, Z2 is an optionally substituted cycloalkyl, such as an optionally substituted cyclobutyl, cyclopentyl, or cyclohexyl. In other embodiments, the cyclobutyl, cyclopentyl, or cyclohexyl, when substituted, is substituted with an alkylamino, such as methylamino or dimethylamino.

[0032] In certain embodiments, Z is

[0033] [ka] It is.

[0034] In certain embodiments, Z 2 is an optionally substituted heterocyclyl, for example an optionally substituted azetidinyl, pyrrolidinyl, or piperidinyl. In other embodiments, the azetidinyl, pyrrolidinyl, or piperidinyl, when substituted, is substituted with alkyl, for example methyl or isopropyl.

[0035] In certain embodiments, Z is

[0036] [ka] It is.

[0037] In certain embodiments, Z is -C(O)NZ 1 Z 2 ;Z 1 and Z 2 are linked together with the N to which they are attached to form an optionally substituted 4-, 5-, or 6-membered heterocycle.

[0038] In certain embodiments, Z is

[0039] [ka] ;R 7 and R 8 are each independently H, CN, halo, alkyl, aminoalkyl, or alkylaminoalkyl; R 9 and R 10 are each independently H, CN, halo, hydroxyl, amino, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl.

[0040] In other embodiments, R 7 and R 8 are each independently H, alkyl, or alkylaminoalkyl; R 9 and R 10are each independently H, alkyl, alkylamino, or alkylaminoalkyl, such as methylamino, dimethylamino, methylaminoalkyl, or dimethylaminoalkyl.

[0041] In certain embodiments, Z is

[0042] [ka] ;X 6 is CH, NH, or N(alkyl); R 11 and R 12 are each independently H, CN, halo, hydroxyl, amino, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl; X 6 When is NH or N(alkyl), R 11 and R 12 is not hydroxyl, amino, or alkylamino. 11 and R 12 are each independently H, CN, alkyl, alkylamino, or alkylaminoalkyl, such as methylamino, dimethylamino, methylaminoalkyl, or dimethylaminoalkyl.

[0043] In certain embodiments, Z is

[0044] [ka] It is.

[0045] In certain embodiments, Z is

[0046] [ka] ;R 13 and R 14 are each independently H, CN, halo, alkyl, aminoalkyl, or alkylaminoalkyl; R 15 and R 16are each independently H, CN, halo, hydroxyl, amino, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl, or R 15 and R 16 are linked to form an optionally substituted 4-, 5-, or 6-membered nitrogen-containing heterocycle.

[0047] In other embodiments, R 13 and R 14 are each independently H, CN, alkyl, aminoalkyl, or alkylaminoalkyl. 15 and R 16 are each independently H, CN, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl, such as methylamino, dimethylamino, methylaminoalkyl, or dimethylaminoalkyl. 13 and R 14 are H and R, respectively. 15 and R 16 are linked to form an optionally substituted 4-, 5-, or 6-membered nitrogen-containing heterocycle.

[0048] In certain embodiments, the optionally substituted 4-, 5-, or 6-membered nitrogen-containing heterocycle is azetidine, pyrrolidine, or piperidine. In other embodiments, the azetidine, pyrrolidine, or piperidine, when substituted, is substituted with alkyl, such as methyl.

[0049] In certain embodiments, Z is

[0050] [ka] It is.

[0051] In certain embodiments, Z is -C(O)NZ 1 Z 2 and Z1 and Z2, together with the N to which they are attached, join to form an optionally substituted heterobicycle.

[0052] In certain embodiments, Z is

[0053] [ka] where R 17 is H, —CN, halo, hydroxyl, amino, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl.

[0054] In certain embodiments, Z is

[0055] [ka] It is.

[0056] In certain embodiments, Z is -NHC(O)NZ 3 Z 4 ;Z 3 and Z 4 are linked together with the N to which they are attached to form an optionally substituted 5-membered heterocycle.

[0057] In certain embodiments, Z is

[0058] [ka] ;R 18 and R 19 are each independently H, CN, halo, alkyl, aminoalkyl, or alkylaminoalkyl; R 20 and R 21 are each independently H, CN, halo, hydroxyl, amino, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl. 20 and R 21 are each independently H or alkylamino. In other embodiments, alkylamino is methylamino or dimethylamino.

[0059] In certain embodiments, Z is

[0060] [ka] It is.

[0061] In certain embodiments, Z is -NHC(O)C(HZ 5 Z 6 ) and Z 5 and Z 6 are linked together with the C to which they are attached to form an optionally substituted 5-membered heterocycle.

[0062] In certain embodiments, Z is

[0063] [ka] ;X 7 is CHR 22 or NR 23 ;R 22 is H, -CN, halo, hydroxyl, amino, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl; R 23 In another embodiment, Z is H, alkyl, alkylamino, or alkylaminoalkyl. 7 CHR 22 ;R 22 is alkylamino, e.g., methylamino or dimethylamino. In other embodiments, X 7 is NR 23 and; R 23 is H or alkyl, such as methyl or isopropyl.

[0064] In certain embodiments, Z is

[0065] [ka] It is.

[0066] In certain embodiments, Z is -NZ 7 Z 8 ;Z 7 and Z 8 are linked together with the N to which they are attached to form an optionally substituted 5- or 6-membered cyclic amide, cyclic urea, or cyclic carbamate.

[0067] In certain embodiments, Z is

[0068] [ka] ;X 8 CR 28 R 29 , N.R. 30 or O;R 24 and R 26 are each independently H or alkyl; R 25 is H or optionally substituted alkyl, aminoalkyl, alkylaminoalkyl, heterocycloalkyl, or heterocyclyl; R 27 is H, or optionally substituted alkyl, amino, alkylamino, aminoalkyl, alkylaminoalkyl, heterocycloalkyl, or heterocyclyl; R 28 and R 29 are each independently H, alkyl, alkylamino, aminoalkyl, or optionally substituted alkylaminoalkyl; R 30 is H, alkyl, alkylaminoalkyl, or optionally substituted cycloalkyl or heterocyclyl; X 8 NR 30 or O, then R 30 is not amino or alkylamino.

[0069] In certain embodiments, X 8 is NH or N-CH3. In another embodiment, X 8 is CH2. In other embodiments, X 8 is O.

[0070] In certain embodiments, R 25 and R 27 is H and R 25 and R 27 The other is,

[0071] [ka] m is 0, 1 or 2; R 30 and R 31 are each independently H or alkyl, or R 30 and R 31 form a carbocyclic ring together with the C to which they are attached; R 32 and R 33 are each independently H, alkyl, hydroxyalkyl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl, or together with the N to which they are attached form an optionally substituted heterocycle.

[0072] In certain embodiments, R 30 and R 31 are each H. In other embodiments, R 30 and R 31 are each alkyl, e.g., methyl.

[0073] In certain embodiments, R 32 and R 33 are each H. In other embodiments, R 32 and R 33 are each alkyl, e.g., methyl.

[0074] In certain embodiments, R 32 is H or CH; R 33is alkyl, hydroxyalkyl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl, such as cyclopropyl, cyclobutyl, azetidinyl, pyrrolidinyl, or piperidinyl. In other embodiments, the cycloalkyl or heterocyclyl, when substituted, is substituted with halo, alkyl, hydroxyl, hydroxyalkyl, or carbamate.

[0075] In certain embodiments, R 32 and R 33 are linked to form an optionally substituted azetidine, pyrrolidine, or piperidine. In other embodiments, the azetidinyl, pyrrolidinyl, or piperidinyl, when substituted, are substituted with halo, alkyl, or hydroxyl.

[0076] In certain embodiments, Z is

[0077] [ka] The file is TIFF2024504927000023.tif90148.

[0078] In certain embodiments, R 25 is H;R 27 is amino or an optionally substituted heterocyclyl. In another embodiment, the optionally substituted heterocyclyl is an optionally substituted oxetanyl, azetidinyl, pyrrolidinyl, piperidinyl, or oxazolidinonyl. In another embodiment, the oxetanyl, azetidinyl, pyrrolidinyl, piperidinyl, or oxazolidinonyl, when substituted, is substituted with alkyl, hydroxyalkyl, or carbamate.

[0079] In certain embodiments, Z is

[0080] [ka] It is.

[0081] In certain embodiments, Z is

[0082] [ka] The file is TIFF2024504927000026.tif61152.

[0083] In certain embodiments, X 8 CR 28 R 29 ;R 28 and R 29 are each independently H, alkyl, alkylamino, aminoalkyl, or optionally substituted alkylaminoalkyl; R 30 is alkyl, alkylaminoalkyl, or optionally substituted cycloalkyl or heterocyclyl. In other embodiments, R 28 and R 29 is H or CH3, and R 28 and R 29 The other is alkylamino, aminoalkyl, or an optionally substituted alkylaminoalkyl. In other embodiments, the alkylaminoalkyl, when substituted, is substituted with hydroxyl.

[0084] In certain embodiments, R 24 , R 25 , R 26 , and R 27 are H, respectively.

[0085] In certain embodiments, X 8 is NR 30 ;R 30is alkylaminoalkyl, or an optionally substituted cycloalkyl or heterocyclyl, such as optionally substituted cyclopropyl, cyclobutyl, oxetanyl, azetidinyl, pyrrolidinyl, or piperidinyl. In another embodiment, cyclopropyl or cyclobutyl, when substituted, is substituted with alkylamino, such as methylamino or dimethylamino. In another embodiment, oxetanyl, azetidinyl, pyrrolidinyl, or piperidinyl, when substituted, is substituted with alkyl, such as methyl.

[0086] In certain embodiments, Z is

[0087] [ka] The file is TIFF2024504927000028.tif43152.

[0088] In certain embodiments, Z is

[0089] [ka] ;X 9 CR 36 R 37 or NR 38 ;X 10 is N or CR 34 ;R 34 and R 35 are each independently H or optionally substituted alkyl; R 36 and R 37 are each independently H, alkyl, alkylamino, aminoalkyl, alkylaminoalkyl, or optionally substituted heterocyclyl; R 38 is alkyl, aminoalkyl or alkylaminoalkyl.

[0090] In other embodiments, Z is

[0091] [ka] In another embodiment, R 36 and R 37 is H or alkyl, and R 36 and R 37 The other is alkylaminoalkyl or an optionally substituted heterocyclyl. In another embodiment, the optionally substituted heterocyclyl is oxetanyl, azetidinyl, pyrrolidinyl, or piperidinyl. In another embodiment, the oxetanyl, azetidinyl, pyrrolidinyl, or piperidinyl, when substituted, is substituted with alkyl.

[0092] In other embodiments, Z is

[0093] [ka] In another embodiment, R 38 is alkylaminoalkyl.

[0094] In certain embodiments, Z is

[0095] [ka] It is.

[0096] In certain embodiments, Z is

[0097] [ka] ;R 39 and R 40 are each independently H or optionally substituted alkyl; R 41 is aminoalkyl, alkylaminoalkyl, or an optionally substituted heterocyclyl or heterocycloalkyl. 41 is aminoalkyl or alkylaminoalkyl. In other embodiments, R 41is an optionally substituted heterocyclyl or heterocycloalkyl. In other embodiments, the heterocyclyl or heterocycloalkyl-substituted alkyl, when substituted, is substituted with alkyl, for example methyl.

[0098] In certain embodiments, Z is

[0099] [ka] It is.

[0100] In certain embodiments, Z is

[0101] [ka] ;R 42 , R 43 , and R 46 are each independently H or optionally substituted alkyl; R 44 , R 45 , and R 47 are each independently H, alkylamino, aminoalkyl, or alkylaminoalkyl. 42 , R 43 , and R 46 are each H. In other embodiments, R 44 , R 45 , and R 47 are each alkylaminoalkyl.

[0102] In certain embodiments, Z is

[0103] [ka] It is.

[0104] In certain embodiments, Z is

[0105] [ka] ;R 48 , R 49 , R 51 , and R 52 are each independently H or optionally substituted alkyl; R 50 and R 53 are each independently H, alkyl, aminoalkyl, or alkylaminoalkyl. 48 , R 49 , R 51 , and R 52 are each H. In other embodiments, R 50 and R 53 are each alkyl.

[0106] In certain embodiments, Z is

[0107] [ka] be.

[0108] In certain embodiments, Z is

[0109] [ka] ;R 54 and R 55 are each independently H, alkyl, aminoalkyl, or alkylaminoalkyl. 54 and R 55 are each alkyl, e.g., methyl.

[0110] In certain embodiments, Z is

[0111] [ka] It is.

[0112] In certain embodiments, Z is

[0113] [ka] ;R 54 is H or optionally substituted alkyl or hydroxyalkyl; R 55 is H, alkyl, aminoalkyl, alkylaminoalkyl, or cycloalkyl. In other embodiments, R 54 is H or hydroxyalkyl. In other embodiments, R 55 is H or cycloalkyl, for example cyclopropyl.

[0114] In certain embodiments, Z is

[0115] [ka] It is.

[0116] Exemplary compounds of formula I are shown in Table 1. It is understood that the compounds in Table 1 include both the free base and the conjugate acid. For example, the compounds in Table 1 may be shown as free base forms, but the compounds as their corresponding complexes or salts with trifluoroacetic acid or hydrochloric acid or salts with other acids are equally within the scope of the present invention. The compounds may be isolated in either the free base form, as a salt (e.g., hydrochloride salt), or both forms. Standard chemical abbreviations may be used in the chemical structures shown below.

[0117] [Table 1] TIFF2024504927000044.tif236170TIFF2024504927000045.tif237170TIFF2024504927000046.tif235170TIF F2024504927000047.tif235170TIFF2024504927000048.tif235170TIFF2024504927000049.tif235170TIFF202 4504927000050.tif235170TIFF2024504927000051.tif235170TIFF2024504927000052.tif235170TIFF2024504 927000053.tif235170TIFF2024504927000054.tif235170TIFF2024504927000055.tif235170TIFF20245049270 00056.tif235170TIFF2024504927000057.tif235170TIFF2024504927000058.tif235170TIFF20245049270000 59.tif235170TIFF2024504927000060.tif235170TIFF2024504927000061.tif235170TIFF2024504927000062.t if235170TIFF2024504927000063.tif235170TIFF2024504927000064.tif235170TIFF2024504927000065.tif23 5170TIFF2024504927000066.tif235170TIFF2024504927000067.tif235170TIFF2024504927000068.tif189170

[0118] In certain embodiments, the compounds of the invention may be prodrugs of compounds of formula I, for example, where a hydroxyl in the parent compound is presented as an ester or carbonate, or a carboxylic acid present in the parent compound is presented as an ester. In certain such embodiments, the prodrug is metabolized in vivo to the active parent compound (e.g., the ester is hydrolyzed to the corresponding hydroxyl or carboxylic acid).

[0119] In certain embodiments, the compounds of the present invention may be racemic. In certain embodiments, the compounds of the present invention may be enriched for one enantiomer. For example, the compounds of the present invention may have more than 30% ee, more than 40% ee, more than 50% ee, more than 60% ee, more than 70% ee, more than 80% ee, more than 90% ee, or more than 95% ee or more. Certain compounds of the present invention have two or more stereocenters. As a result, such compounds can be enriched for one or more diastereomers. For example, the compounds of the present invention may have more than 30% de, more than 40% de, more than 50% de, more than 60% de, more than 70% de, more than 80% de, more than 90% de, or more than 95% de or more.

[0120] In certain embodiments, as described in detail below, the present invention relates to a method of treating or preventing cancer or inflammatory disorders with a compound of formula I, or a pharma- ceutically acceptable salt thereof. In certain embodiments, the administered therapeutic preparation may be enriched to provide predominantly one enantiomer of a compound (e.g., a compound of formula I). ​​An enantiomerically enriched mixture may, for example, contain at least 60 mol percent of one enantiomer, or more preferably at least 75, 90, 95, or even 99 mol percent of one enantiomer. In certain embodiments, a compound enriched for one enantiomer is substantially free of the other enantiomer, where substantially free means that the substance of interest constitutes less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1% when compared to the amount of the other enantiomer in, for example, a composition or compound mixture. For example, if a composition or compound mixture contains 98 grams of a first enantiomer and 2 grams of a second enantiomer, it would be said to contain 98 mole percent of the first enantiomer and only 2% of the second enantiomer.

[0121] In certain embodiments, a therapeutic preparation may be enriched to provide predominantly one diastereomer of a compound (e.g., of a compound of Formula I). ​​A diastereomerically enriched mixture can contain, for example, at least 60 mol percent of one diastereomer, or more preferably at least 75, 90, 95, or even 99 mol percent of one diastereomer.

[0122] In certain embodiments, the present invention provides a pharmaceutical preparation suitable for use in human patients in the treatment of cancer, inflammatory disorders, comprising an effective amount of any compound of formula I and one or more pharma- ceutically acceptable excipients. In certain embodiments, the pharmaceutical preparation may be for use in the treatment or prevention of conditions or diseases described herein. In certain embodiments, the pharmaceutical preparation has sufficiently low pyrogenic activity to be suitable for use in human patients.

[0123] Any of the compounds of the above structure can be used in the manufacture of a medicament for the treatment of any of the diseases or conditions disclosed herein.

[0124] The present invention is also intended to include all isotopes of atoms occurring in the compounds disclosed herein. Isotopes include those atoms having the same atomic number but different mass numbers. By way of general example and without limitation, isotopes of hydrogen include deuterium (D). Throughout this application, any designation of carbon in a structure, when used without further designation, is: 12 C. 13 C, or 14 It is intended to represent all isotopes of carbon, such as C. Throughout this application, when any designation for hydrogen in a structure is used without further designation, 1 H, 2 H, or 3It is intended to represent all isotopes of hydrogen, such as H. Isotopically labeled compounds can generally be prepared by conventional techniques known to those of skill in the art, substituting the appropriate isotopically labeled reagent for the non-labeled reagent utilized.

[0125] Exemplary compounds of formula I are shown in Examples 1-93 and Tables 101-137. It is understood that the compounds in Examples 1-93 and Tables 101-137 include both the free base and the conjugate acid. For example, the compounds in Examples 1-93 and 101-137 may be shown as complexes or salts with trifluoroacetic acid or hydrochloric acid, but the compounds in their corresponding free base form or as salts with other acids are equally within the scope of the present invention. The compounds may be isolated in either the free base form, as a salt (e.g., hydrochloride salt), or both forms. Standard chemical abbreviations may be used in the chemical structures shown below.

[0126] II. Use of MK2 inhibitors In certain aspects, the invention provides methods for treating or preventing disorders associated with mitogen-activated protein kinase-activated protein kinase-2 (MK2), comprising administering to a subject, for example, a therapeutically effective amount of a compound of formula I.

[0127] In certain embodiments, the MK2 associated disorder is an inflammatory disorder, hi other embodiments, the MK2 associated disorder is cancer.

[0128] In certain embodiments, the present invention provides methods of treating cancer, comprising administering to a subject, for example, a therapeutically effective amount of a compound of formula I.

[0129] In certain embodiments, the cancer is acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), adrenocortical carcinoma, AIDS-related cancers (Kaposi's sarcoma and lymphoma), anal cancer, appendix cancer, atypical terato / rhabdoid tumor, basal cell carcinoma, bile duct cancer (including extrahepatic), bladder cancer, bone cancer (including osteosarcoma and malignant fibrous histiocytoma), brain tumors (astrocytoma, brain and spinal cord tumors, brain stem glioma, central nervous system atypical terato / rhabdoid tumor, central nervous system embryonal tumor, craniopharyngioma, ependymoblastoma, ependymoma, medulloblastoma, medulloepithelioma, pineal parenchymal tumor of intermediate differentiation, supratentorial tumor ... primitive neuroectodermal tumor and pineoblastoma, etc.), breast cancer, bronchial tumor, Burkitt's lymphoma, basal cell carcinoma, bile duct cancer (including extrahepatic), bladder cancer, bone cancer (including osteosarcoma and malignant fibrous histiocytoma), carcinoid tumor, carcinoma of unknown primary, central nervous system (atypical teratoid tumor / rhabdoid tumor, embryonal tumor and lymphoma, etc.), cervical cancer, childhood cancer, chordoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myeloproliferative disorder, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma (mycosis fungoides and Sézary syndrome), bile duct, bile (extrahepatic), non Invasive Ductal Carcinoma (DCIS), Germ Cell Tumors (Central Nervous System), Endometrial Cancer, Ependymoblastoma, Ependymoma, Esophageal Cancer, Esthesioneuroblastoma, Ewing's Sarcoma Family of Tumors, Extracranial Germ Cell Tumors, Extragonadal Germ Cell Tumors, Extrahepatic Bile Duct Cancer, Eye Cancer (Intraocular Melanoma, Retinoblastoma), Fibrous Histiocytoma of Bone (Including Malignant and Osteosarcoma), Gallbladder Cancer, Gastric (Stomach) Cancer, Gastrointestinal Carcinoid Tumor, Gastrointestinal Mesenchymal Tumor (GIST), Germ Cell Tumors (Extracranial, Extragonadal, Ovarian), Gestational Trophoblastic Tumor, Glioblastoma, Glioma, Hairy Cell Leukemia, Head and Neck Cancer, Heart Cancer, Hepatocellular (Liver) ) cancer, histiocytosis, Langerhans cell, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumors (endocrine, pancreatic), Kaposi's sarcoma, kidney (including renal cell), Langerhans cell histiocytosis, laryngeal cancer, leukemia (including acute lymphoblastic (ALL), acute myeloid (AML), chronic lymphocytic (CLL), chronic myeloid (CML), hairy cell), lip and oral cavity cancer, liver cancer (primary), lobular carcinoma in situ (LCIS), lung cancer (non-small cell and small cell), lymphoma (AIDS-related, Burkitt, cutaneous T cell (mycosis fungoides and Sézary syndrome), Hodgkin,Non-Hodgkin's, Primary Central Nervous System (CNS), Male Breast Cancer, Malignant Fibrous Histiocytoma and Osteosarcoma of Bone, Medulloblastoma, Melanoma (including Intraocular (Eye)), Merkel Cell Carcinoma, Mesothelioma (Malignant), Metastatic Squamous Cell Neck Cancer of Unknown Primary, Midline Tract Carcinoma Involving the NUT Gene, Oral Cancer, Multiple Endocrine Neoplasia Syndrome, Multiple Myeloma / Plasma Cell Neoplasm, Mycosis Fungoides, Myelodysplastic Syndrome, Myelodysplastic / Myeloproliferative Neoplasm, Chronic Myeloid Leukemia (CML), Acute Myeloid Leukemia (AML), Myeloma and Multiple Myeloma, Myeloproliferative Disorders (Chronic), Nasal Cavity and Paranasal Sinus Cancer, Nasopharyngeal Cancer, Neuroblastoma, Non-Hodgkin's Lymphoma, Non-Small Cell Lung Cancer, Oral Cancer, Oral Cavity Cancer, Lip and Opharyngeal Cancer cancer, osteosarcoma and malignant fibrous histiocytoma of bone, ovarian cancer (including epithelial, germ cell, and low malignant potential tumors), pancreatic cancer (including islet cell tumors), papilloma, paraganglioma, paranasal sinus and nasal cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, intermediately differentiated pineal parenchymal tumor, pineoblastoma and supratentorial primitive neuroectodermal tumor, pituitary tumor, plasma cell neoplasm / multiple myeloma, pleuropulmonary blastoma, pregnancy and breast cancer, primary central nervous Central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, renal cell (kidney) cancer, renal pelvis and ureter, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma (Ewing sarcoma tumor family, Kaposi, soft tissue, uterine, etc.), Sézary syndrome, skin cancer (melanoma, Merkel cell carcinoma, non-melanoma, etc.), small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, squamous cell neck cancer of unknown primary site, metastatic gastric (stomach) cancer, supratentorial It may be one or a variant of the following: primitive neuroectodermal tumor, T-cell lymphoma (skin, mycosis fungoides and Sézary syndrome), testicular cancer, laryngeal cancer, thymoma and thymic cell carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastic tumor (gestational), unknown primary, unusual cancer of childhood, ureter and renal pelvis, transitional cell carcinoma, urethral cancer, uterine cancer, endometrium, uterine sarcoma, Waldenstrom's macroglobulinemia and Wilms' tumor.

[0130] In certain embodiments, the cancer is a KRAS or BRAF dependent cancer.

[0131] In certain embodiments, the cancer is a solid tumor. The subject is generally one who has been diagnosed with a cancerous tumor or who has previously been treated for a cancerous tumor (e.g., the tumor has previously been removed by surgery). The cancerous tumor may be a primary tumor and / or a secondary (e.g., metastatic) tumor.

[0132] In certain embodiments, the subject is a mammal, such as a human.

[0133] In certain embodiments, the cancer is associated with bladder, bone marrow, breast, colon, kidney, liver, lung, ovary, pancreas, prostate, skin, or thyroid tissue.

[0134] In certain embodiments, the method of treating cancer further comprises conjointly administering radiation therapy.

[0135] In some embodiments, the methods of treating cancer further comprise co-administering one or more additional chemotherapeutic agents.

[0136] Chemotherapeutic agents that may be administered in conjunction with the compounds of the invention include ABT-263, aminoglutethimide, amsacrine, anastrozole, asparaginase, AZD5363, bacille Calmette-Guerin vaccine (bcg), bicalutamide, bleomycin, bortezomib, buserelin, busulfan, campothecin, capecitabine, carboplatin, carfilzomib, carmustine, chlorambucil, chloroquine, cisplatin, cladribine, clodronate, cobimetinib, colchicine, cyclophosphamide, cyprotease, cyclosporine ... cerebrospinal fluid, cytarabine, dacarbazine, dactinomycin, daunorubicin, demethoxyviridin, dexamethasone, dichloroacetic acid, dienestrol, diethylstilbestrol, docetaxel, doxorubicin, epirubicin, estradiol, estramustine, etoposide, everolimus, exemestane, filgrastim, fludarabine, fludrocortisone, fluorouracil and 5-fluorouracil, fluoxymesterone, flutamide, gemcitabine, genistein, goserelin, hydroxyurea, idarubicin, ifosfamidis , imatinib, interferon, irinotecan, lenalidomide, letrozole, leucovorin, leuprolide, levamisole, lomustine, lonidamine, LY2603618, mechlorethamine, medroxyprogesterone, megestrol, melphalan, mercaptopurine, mesna, metformin (e.g., metformin HCl), methotrexate, miltefosine, mitomycin, mitotane, mitoxantrone, MK2206, nilutamide, nocodazole, octreotide, oxaliplatin, paclitaxel, pamidronate, pentosine, tatin, pazopanib, perifosine, PF-04691502, PF477736, plicamycin, pomalidomide, porfimer, procarbazine, raltitrexed, rituximab, romidepsin, selumetinib, sorafenib, streptozocin, sunitinib, suramin, tamoxifen, temozolomide, temsirolimus, teniposide, testosterone, thalidomide, thioguanine, thiotepa, titanocene dichloride, topotecan, trametinib, trastuzumab, tretinoin, vinblastine, vincristine, vindesine, vinorelbine,and vorinostat (SAHA).

[0137] For example, chemotherapeutic agents that may be administered in conjunction with the compounds of the invention include aminoglutethimide, amsacrine, anastrozole, asparaginase, bcg, bicalutamide, bleomycin, bortezomib, buserelin, busulfan, campothecin, capecitabine, carboplatin, carfilzomib, carmustine, chlorambucil, chloroquine, cisplatin, cladribine, clodronate, colchicine, cyclophosphamide, cyproterone, cytarabinose, and cyclosporine. bin, dacarbazine, dactinomycin, daunorubicin, demethoxyviridin, dichloroacetic acid, dienestrol, diethylstilbestrol, docetaxel, doxorubicin, epirubicin, estradiol, estramustine, etoposide, everolimus, exemestane, filgrastim, fludarabine, fludrocortisone, fluorouracil, fluoxymesterone, flutamide, gemcitabine, genistein, goserelin, hydroxyurea, idarubicin, ifo Sulfamide, imatinib, interferon, irinotecan, lenalidomide, letrozole, leucovorin, leuprolide, levamisole, lomustine, lonidamine, mechlorethamine, medroxyprogesterone, megestrol, melphalan, mercaptopurine, mesna, metformin (e.g., metformin HCl), methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, nocodazole, octreotide, oxaliplatin, paclitaxel, pami These include doronate, pentostatin, perifosine, plicamycin, pomalidomide, porfimer, procarbazine, raltitrexed, rituximab, sorafenib, streptozocin, sunitinib, suramin, tamoxifen, temozolomide, temsirolimus, teniposide, testosterone, thalidomide, thioguanine, thiotepa, titanocene dichloride, topotecan, trastuzumab, tretinoin, vinblastine, vincristine, vindesine, and vinorelbine. In certain embodiments, the chemotherapeutic agent is cisplatin. In certain embodiments, the additional chemotherapeutic agent is a CHK1 inhibitor. In certain embodiments, the additional chemotherapeutic agent is an alkylating agent.

[0138] "PF477736" refers to at least the following construct commercially available from Sigma-Aldrich (catalog number PZ0186):

[0139] [ka]

[0140] "LY2603618" refers to at least the following construct commercially available from Sigma-Aldrich (catalog number SML2855):

[0141] [ka]

[0142] Many combination therapies have been developed for cancer treatment. In certain embodiments, the compound of the present invention can be administered in combination with combination therapy. Examples of combination therapy that the compound of the present invention can be administered in combination with are listed in Table 2.

[0143] [Table 2] TIFF2024504927000072.tif255155TIFF2024504927000073.tif255152TIFF2024504927000074.tif255152 TIFF2024504927000075.tif255156TIFF2024504927000076.tif255156TIFF2024504927000077.tif109168

[0144] In some embodiments, the co-administered chemotherapeutic agent is an immuno-oncology therapeutic agent, such as an inhibitor of CTLA-4, indoleamine-2,3-dioxygenase, and / or PD-1 / PD-L1.

[0145] In certain embodiments, administering the MK2 inhibitor(s) of formula I in combination with one or more additional therapeutic agents (e.g., one or more additional chemotherapeutic agents) results in improved efficacy compared to administering the MK2 inhibitor (e.g., a compound of formula I) or each of the one or more additional therapeutic agents individually. In certain such embodiments, administering together results in an additive effect, where additive effect refers to the sum of the effects of administering the MK2 inhibitor and the one or more additional therapeutic agents individually. In certain embodiments, coadministration results in a synergistic effect.

[0146] In certain embodiments, the MK2 inhibitor and the one or more additional chemotherapeutic agents are administered simultaneously, hi alternative embodiments, the one or more additional chemotherapeutic agents are administered within about 5 minutes to about 168 hours before or after administration of the MK2 inhibitor.

[0147] In certain embodiments, the present invention relates to a method of inhibiting cancerous cell proliferation, comprising contacting a cancerous cell with an effective amount of a compound of formula I.

[0148] The present invention also relates to a method of inhibiting MK2 activity in a cell, comprising contacting the cell with a compound of formula I. In certain embodiments, the cell is a cancer cell. Such methods can be performed in vivo or in vitro.

[0149] The present invention also relates to a method of treating or preventing a metabolic disorder, comprising administering to a subject a compound of formula I. In certain embodiments, the metabolic disorder is diabetes, insulin resistance, obesity, or metabolic syndrome. In some embodiments, the diabetes is type I, type II, or gestational diabetes. In some embodiments, the treatment or prevention affects glycogenolysis or gluconeogenesis in the subject. In some embodiments, the treatment or prevention reduces hepatic glucose production, hyperglycemia, hepatic steatosis, insulin resistance, inflammation associated with insulin resistance, dyslipidemia associated with insulin resistance, or any combination thereof in the subject.

[0150] For example, in certain embodiments of the method of treating diabetes, the method further comprises the co-administration of one or more additional antidiabetic agents. Antidiabetic agents that may be administered in combination with the compounds of the invention include, but are not limited to, sulfonylureas, biguanides, alpha-glucosidase inhibitors, thiazolidinediones (TZDs), dipeptidyl peptidase inhibitors (DPP-4 inhibitors), non-sulfonylurea insulin secretagogues, glucagon-like peptide 1 analogs (GLP-1 analogs), and insulin. More particularly, antidiabetic agents include, but are not limited to, metformin (metformin HCl), glyburide, glimepiride, glipiride, glipizide, chlorpropamide, gliclazide, acarbose, miglitol, pioglitazone, troglitazone, rosiglitazone, isaglitazone, muraglitizar, peliglitazar, sitagliptin, saxagliptin, vildagliptin, alogliptin, linagliptin, dutogliptin, dutogliptin, repaglinide, nateglinide, mitiglindine, exenatide, liraglutide, albiglutide, and insulin. In certain preferred embodiments, the one or more additional antidiabetic agents comprises metformin.

[0151] III. Pharmaceutical Compositions In certain embodiments, the present invention provides a pharmaceutical composition comprising a compound of formula I and a pharma- ceutically acceptable carrier.

[0152] The compositions and methods of the present invention can be used to treat an individual in need thereof. In certain embodiments, the individual is a mammal, such as a human or a non-human mammal. When administered to an animal, such as a human, the compositions or compounds are preferably administered as a pharmaceutical composition, for example comprising the compound of the present invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers, such as aqueous solutions, such as water or physiologically buffered saline, or other solvents or vehicles, such as glycols, glycerol, oils, such as olive oil or injectable organic esters, are well known in the art. In a preferred embodiment, if such pharmaceutical compositions are for administration to humans, particularly for invasive routes of administration (i.e., routes such as injection or implantation that avoid transport or diffusion through epithelial barriers), the aqueous solutions are pyrogen-free or substantially pyrogen-free. Excipients can be selected, for example, to effect delayed release of the drug or to selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in unit dosage form such as tablets, capsules (such as sprinkle capsules and gelatin capsules), granules, lyophilized products to be reconstituted, powders, solutions, syrups, suppositories, injections, etc. The composition can also be in a transdermal delivery system, such as a skin patch. The composition can also be in a solution suitable for external administration, such as eye drops.

[0153] Pharmaceutically acceptable carriers may contain physiologically acceptable agents that act, for example, to stabilize, increase solubility, or increase absorption of compounds such as the compounds of the present invention. Such physiologically acceptable agents include carbohydrates such as glucose, sucrose, or dextran, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins, or other stabilizers or excipients. The choice of pharma-ceutically acceptable carriers, including physiologically acceptable agents, depends, for example, on the route of administration of the composition. The pharmaceutical composition preparation may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) may also be, for example, a liposome or other polymer matrix that can incorporate the compounds of the present invention therein. Liposomes, including, for example, phospholipids or other lipids, are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively simple to prepare and administer.

[0154] The phrase "pharmaceutical acceptable" is used herein to refer to those compounds, materials, compositions and / or dosage forms that are suitable, within the limits of sound medical judgment, for use in contact with the tissues of humans and animals, without causing excessive toxicity, hypersensitivity, allergic response or other problem or complication, and commensurate with a reasonable benefit / risk ratio.

[0155] The phrase "pharmaceutical acceptable carrier" as used herein means a pharma- ceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the patient. Some examples of materials that can serve as pharma- ceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and cellulose derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) soluble oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil. (10) oils, (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances used in pharmaceutical preparations.

[0156] The pharmaceutical compositions (preparations) can be administered to a subject by any of a number of routes of administration, including, for example, orally (e.g., drenches, such as aqueous or non-aqueous solutions or suspensions, tablets, capsules (such as sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes applied to the tongue, etc.); absorption through the oral mucosa (e.g., sublingually); anally, rectally, or vaginally (e.g., as a pessary, cream, or foam); parenterally (e.g., intramuscularly, intravenously, subcutaneously, or intrathecally, e.g., as a sterile solution or suspension); intranasally; intraperitoneally; subcutaneously; transdermally (e.g., as a patch applied to the skin); and topically (e.g., as a cream, ointment, or spray applied to the skin, or as eye drops). The compounds can also be formulated for inhalation. In certain embodiments, the compounds can simply be dissolved or suspended in sterile water. Details of suitable routes of administration and compositions suitable therefor can be found, for example, in U.S. Pat. Nos. 6,110,973, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, and in the patents cited therein.

[0157] The preparations may conveniently be in unit dosage form and may be prepared by any method known in the pharmaceutical art. The amount of active ingredient that can be mixed with carrier materials to produce a single dosage form will vary depending on the host being treated, the individual mode of administration. The amount of active ingredient that can be mixed with carrier materials to produce a single dosage form will generally be the amount of compound that produces a therapeutic effect. Generally, out of 100%, this amount will be within the range of about 1% to about 99% of active ingredient, preferably about 5% to about 70%, most preferably about 10% to about 30%.

[0158] Methods of preparing these formulations or compositions include the step of bringing into association an active compound, such as a compound of the present invention, with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0159] Formulations of the present invention suitable for oral administration may be in the form of capsules (such as sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (with a flavored base, usually sucrose and acacia or tragacanth), lyophilisates, powders, granules, or as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as a pastille (with an inert base such as gelatin and glycerin or sucrose and acacia) and / or as a mouthwash, etc. The composition or compound may also be administered as a bolus, electuary, or paste.

[0160] To prepare solid dosage forms for oral administration (such as capsules (such as sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules, and the like), the active ingredient is mixed with one or more pharma- ceutically acceptable carriers, such as sodium citrate or dicalcium phosphate and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants, such as glycerol. (4) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; (5) dissolution retarders such as paraffin; (6) absorption accelerators such as quaternary ammonium compounds; (7) wetting agents such as cetyl alcohol and glycerol monostearate; (8) absorbents such as kaolin and bentonite clay; (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof; (10) complexing agents such as modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (such as sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical compositions may also include buffering agents. Similar types of solid compositions may also be used as fillers for soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols and the like.

[0161] Tablets can be made by compression or molding, optionally with one or more accessory ingredients.Compressed tablets can be prepared using binders (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surface active agents or dispersants.Molded tablets can be made by molding a mixture of the powdered compound moistened with an inert liquid diluent in a suitable machine.

[0162] Tablets and other solid dosage forms of pharmaceutical compositions, such as dragees, capsules (such as sprinkle capsules and gelatin capsules), pills and granules, can be optionally scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the art of pharmaceutical compounding. They can also be formulated to provide slow or controlled release of the active ingredient, for example using hydroxypropylmethylcellulose in various proportions to provide the desired release profile, other polymer matrices, liposomes and / or microspheres. They can be sterilized, for example by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water or some other sterile injectable medium immediately before use. These compositions can also optionally contain opacifying agents, and can be compositions that release the active ingredient(s) only, or preferentially, in a certain part of the gastrointestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient can also be in microencapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0163] The liquid dosage form useful for oral administration includes pharma- ceutically acceptable emulsion, reconstituted lyophilisate, microemulsion, solution, suspension, syrup and elixir.In addition to active ingredient, liquid dosage form can contain inert diluent commonly used in the art, such as water or other solvent, cyclodextrin and its derivatives, solubilizer and emulsifier, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oil (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol and fatty acid ester of sorbitan, and mixtures thereof.

[0164] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0165] Suspensions may contain, in addition to the active compound, suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, tragacanth, and mixtures thereof.

[0166] Formulations of pharmaceutical compositions for rectal, vaginal or urethral administration can be present as suppositories, which can be prepared by mixing one or more active compounds with one or more suitable non-irritating excipients or carriers including, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, which are solid at room temperature but liquid at body temperature and thereby melt in the rectum or vaginal cavity to release the active compounds.

[0167] Formulations of the pharmaceutical composition for administration to the mouth may be present as a mouthwash or an oral spray or an oral ointment.

[0168] Alternatively or additionally, the compositions may be formulated for delivery via a catheter, stent, wire or other intraluminal device, which may be particularly useful for delivery to the bladder, urethra, ureter, rectum or intestine.

[0169] Formulations which are suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.

[0170] Dosage forms for external or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants.The active compound may be mixed under sterile conditions with a pharma- ceutical acceptable carrier and any preservatives, buffers or propellants that may be required.

[0171] The ointments, pastes, creams and gels may contain, in addition to the active compound, excipients such as animal and vegetable fats, oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, salicylic acid, talc and zinc oxide, or mixtures thereof.

[0172] Powders and sprays can contain, in addition to the active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances.In addition, sprays can contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0173] Transdermal patch has the additional advantage of providing controlled delivery of the compound of the present invention to the body.Such dosage forms can be prepared by dissolving or dispersing active compound in suitable medium.Absorption enhancers can also be used to increase the permeation of the compound through the skin.The rate of such permeation can be controlled by either providing a rate-controlling membrane or dispersing the compound in a polymer matrix or gel.

[0174] Ophthalmic formulations, eye ointments, powders, solutions, etc. are also contemplated within the scope of the present invention. Exemplary ophthalmic formulations are described in U.S. Patent Application Publication Nos. 2005 / 0080056, 2005 / 0059744, 2005 / 0031697, and 2005 / 004074, the contents of which are incorporated herein by reference, and U.S. Patent No. 6,583,124. If desired, liquid ophthalmic formulations have similar properties to tears, aqueous humor, or vitreous humor, or are compatible with such fluids. The preferred route of administration is topical administration (e.g., external administration such as eye drops or administration via implant).

[0175] The phrases "parenteral administration" and "administered parenterally" as used herein refer to modes of administration other than enteral and topical administration, usually by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, intrathecal, intraspinal and intrasternal injection and infusion.The pharmaceutical composition suitable for parenteral administration comprises one or more active compounds in combination with one or more pharmacologic acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders, which can be reconstituted into sterile injectable solutions or dispersions immediately before use, and can contain antioxidants, buffers, bacteriostatic agents, solutes that make the formulation isotonic with the blood of intended recipient, or suspending or thickening agents.

[0176] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained by using coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by using surfactants.

[0177] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifiers and dispersing agents. Prevention of microbial action may be ensured by the inclusion of various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol sorbic acid, etc. It may be desirable to include isotonic agents such as sugars, sodium chloride, etc. in the composition. In addition, prolonged absorption of injectable pharmaceutical forms may be brought about by the inclusion of agents that delay absorption, such as aluminum monostearate and gelatin.

[0178] In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be accomplished by using a liquid suspension of crystalline or amorphous material with poor water solubility. In this case, the absorption rate of the drug depends on the dissolution rate, which may also depend on the crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

[0179] Injectable depot forms are made by forming microencapsulated matrices of the target compound in biodegradable polymers such as polylactide-polyglycolide.The rate of drug release can be controlled depending on the ratio of drug to polymer and the properties of the particular polymer used.Other examples of biodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

[0180] When used in the methods of the invention, the active compounds may be administered as is or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably 0.5 to 90%) of the active ingredient in combination with a pharma- ceutically acceptable carrier.

[0181] The introduction method can also be provided by rechargeable or biodegradable devices.Various delayed release polymeric devices, including proteinaceous biologics, have been developed and tested in vivo in recent years for controlled drug delivery.Various biocompatible polymers, including both biodegradable and non-degradable polymers (such as hydrogels), can be used to form implants for sustained release of compounds at individual target sites.

[0182] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied to obtain an amount of the active ingredient effective to achieve a desired therapeutic response for a particular patient, administration composition and mode without causing toxicity to the patient.

[0183] The selected dosage level will depend on a variety of factors, such as the activity of the particular compound, the combination of compounds or their esters, salts or amides used, the route of administration, the time of administration, the rate of excretion of the particular compound(s) used, the duration of treatment, other drugs, compounds and / or materials used in combination with the particular compound(s) used, the age, sex, weight, illness, general health, past medical history of the patient being treated, and similar factors well known in the medical arts.

[0184] A physician or veterinarian of ordinary skill in the art can readily determine and prescribe the therapeutically effective amount of the pharmaceutical composition required. For example, a physician or veterinarian can begin the dosage of the pharmaceutical composition or compound at a level lower than that required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved. A "therapeutically effective amount" refers to an amount of a compound sufficient to induce the desired therapeutic effect. It is generally understood that the effective amount of a compound varies according to the subject's weight, sex, age, and medical history. Other factors that affect the effective amount include, but are not limited to, the severity of the patient's illness, the disorder being treated, the stability of the compound, and other types of therapeutic agents administered together with the compound of the present invention, if desired. A larger total dose can be delivered by repeated administration of the agent. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13 ed., 1814-1882, incorporated herein by reference).

[0185] In general, a suitable daily dose of an active compound used in the compositions and methods of the present invention will be that amount of the compound that is the minimum dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.

[0186] If desired, the effective daily dose of active compound can be administered separately at appropriate intervals throughout the day, as 1, 2, 3, 4, 5, 6 or more divided doses, optionally in unit dosage form.In certain embodiments of the present invention, active compound can be administered 2 or 3 times a day.In a preferred embodiment, active compound is administered once a day.

[0187] The patients to be treated are primates, particularly humans, and other mammals, such as horses, cattle, pigs and sheep, and generally any animal in need, such as poultry and pets.

[0188] In certain embodiments, the compounds of the present invention may be used alone or may be administered in combination with another type of therapeutic agent. As used herein, the phrase "administered in combination" refers to any form of administration of two or more different therapeutic compounds, such that the second compound is administered while the previously administered therapeutic compound is still effective on the body. (For example, the two compounds are effective on the patient at the same time, which may include a synergistic effect of the two compounds). For example, different therapeutic compounds may be administered either simultaneously or sequentially, either as the same formulation or as separate formulations. In certain embodiments, different therapeutic compounds may be administered within 1 hour, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, or 1 week of each other. Thus, an individual undergoing such treatment may benefit from the combined effect of different therapeutic compounds.

[0189] In certain embodiments, administering a compound of the invention in combination with one or more additional therapeutic agents (e.g., one or more additional chemotherapeutic agents) results in improved efficacy compared to administration of a compound of the invention (e.g., a compound of Formula I) or each of the one or more additional therapeutic agents individually. In certain such embodiments, administering together results in an additive effect, where additive effect refers to the sum of the effects of each of the administration of a compound of the invention and one or more additional therapeutic agents individually.

[0190] The present invention includes the use of pharma- ceutically acceptable salts of the compounds of the present invention in the compositions and methods of the present invention. As used herein, the term "pharma- ceutically acceptable salts" includes salts derived from inorganic or organic acids, including, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, phosphoric acid, formic acid, acetic acid, lactic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, glycolic acid, salicylic acid, citric acid, methanesulfonic acid, benzenesulfonic acid, benzoic acid, malonic acid, trifluoroacetic acid, trichloroacetic acid, naphthalene-2-sulfonic acid, and other acids. Pharmaceutically acceptable salt forms may include forms in which the ratio of the molecules that make up the salt is not 1:1. For example, a salt may contain more than one inorganic or organic acid molecule per base molecule, such as two hydrochloric acid molecules per molecule of the compound of formula I. As another example, a salt may contain less than one inorganic or organic acid molecule per base molecule, such as two compounds of formula I molecules per molecule of tartaric acid.

[0191] In further embodiments, contemplated salts of the present invention include, but are not limited to, alkyl, dialkyl, trialkyl, or tetraalkyl ammonium salts. In certain embodiments, contemplated salts of the present invention include, but are not limited to, L-arginine, benenthamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, contemplated salts of the present invention include, but are not limited to, Na, Ca, K, Mg, Zn, or other metal salts.

[0192] Pharmaceutically acceptable acid addition salts can also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, etc. Mixtures of such solvates can also be prepared. The source of such solvates can be from the solvent of crystallization, either inherent in the solvent of preparation or crystallization, or adventitious to such solvent.

[0193] Wetting agents, emulsifying agents and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition.

[0194] Examples of pharma- ceutically acceptable antioxidants include: (1) water-soluble antioxidants such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium disulfite, and sodium sulfite; (2) fat-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, and α-tocopherol; and (3) metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, and phosphoric acid.

[0195] definition The term "acyl" is art recognized and refers to a group represented by the general formula: hydrocarbylC(O)-, preferably alkylC(O)-.

[0196] The term "acylamino" is art recognized and refers to an amino group substituted with an acyl group and may be represented, for example, by the formula: hydrocarbylC(O)NH--.

[0197] The term "acyloxy" is art recognized and refers to a group represented by the general formula: hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0198] The term "alkoxy" refers to an alkyl group, preferably a lower alkyl group, having an oxygen attached thereto. Representative alkoxy groups include methoxy, -OCF3, ethoxy, propoxy, tert-butoxy, and the like.

[0199] The term "cycloalkyloxy" refers to a cycloalkyl group having an oxygen attached thereto.

[0200] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and can be represented by the general formula: alkyl-O-alkyl.

[0201] The term "alkylaminoalkyl" refers to an alkyl group substituted with an alkylamino group.

[0202] The term "alkenyl" as used herein means an aliphatic group containing at least one double bond, and is intended to include both "unsubstituted alkenyl" and "substituted alkenyl", the latter of which refers to an alkenyl moiety having a substituent replacing a hydrogen on one or more carbons of the alkenyl group. Such a substituent may be present on one or more carbons that are included or not included in one or more double bonds. In addition, such substituents include all of those considered for alkyl groups, as discussed below, except where stability would be prohibitive. For example, substitution of an alkenyl group with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is considered.

[0203] An "alkyl" group or "alkane" is a straight or branched chain non-aromatic hydrocarbon that is fully saturated. Typically, a straight or branched chain alkyl group has from 1 to about 20 carbon atoms, preferably from 1 to about 10 carbon atoms, unless otherwise specified. Examples of straight and branched chain alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl and octyl. C1-C6 straight or branched chain alkyl groups are also referred to as "lower alkyl" groups.

[0204] Furthermore, the term "alkyl" (or "lower alkyl"), as used throughout the specification, examples, and claims, is intended to include both "unsubstituted alkyl" and "substituted alkyl," the latter of which refers to an alkyl moiety having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. Such substituents, unless otherwise specified, include, for example, halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, where appropriate. For example, substituents of substituted alkyl include substituted and unsubstituted amino, azido, imino, amido, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamides, sulfamoyl and sulfonates), and silyl groups, as well as ethers, alkylthio, carbonyl (including ketones, aldehydes, carboxylates, and esters), -CF3, -CN, and the like. Exemplary substituted alkyls are described below. Cycloalkyls can be further substituted with alkyls, alkenyls, alkoxys, alkylthios, aminoalkyls, carbonyl-substituted alkyls, -CF3, -CN, and the like.

[0205] When used in connection with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, the term "C x-y " means to include groups containing x to y carbons in the chain. For example, the term "C x-y"Alkyl" means a substituted or unsubstituted saturated hydrocarbon group, including straight-chain and branched-chain alkyl groups containing x to y carbons in the chain, and includes haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl. CO alkyl refers to a hydrogen when the group is in a terminal position and to a bond when the group is internal. The term "C 2-y alkenyl" and "C 2-y "Alkynyl" refers to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one double or triple bond respectively.

[0206] The term "alkylamino," as used herein, means an amino group substituted with at least one alkyl group.

[0207] The term "alkylthio," as used herein, refers to a thiol group substituted with an alkyl group and can be represented by the general formula: alkyl-S-.

[0208] The term "alkynyl" as used herein means an aliphatic group containing at least one triple bond, and is intended to include both "unsubstituted alkynyl" and "substituted alkynyl", the latter of which refers to an alkynyl moiety having a substituent replacing a hydrogen on one or more carbons of the alkynyl group. Such a substituent may be present on one or more carbons that are included or not included in one or more triple bonds. In addition, such substituents include all of those considered for alkyl groups, as discussed above, except where stability would be disruptive. For example, substitution of an alkynyl group with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is considered.

[0209] The term "amide" as used herein refers to the group:

[0210] [ka]

[0211] wherein each R independently represents hydrogen or a hydrocarbyl group, or two R together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure. means.

[0212] The terms "amine" and "amino" are art recognized and refer to both unsubstituted and substituted amines, and salts thereof, such as:

[0213] [ka]

[0214] wherein each R independently represents hydrogen or a hydrocarbyl group, or two R together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure. "R" means a moiety that can be represented by the formula:

[0215] The term "aminoalkyl," as used herein, refers to an alkyl group substituted with an amino group.

[0216] The term "aralkyl," as used herein, refers to an alkyl group substituted with an aryl group.

[0217] The term "aryl" as used herein includes substituted or unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon. Preferably, the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to the two connected rings, where at least one of the rings is aromatic and the other cyclic ring can be, for example, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

[0218] The term "carbamate" is art recognized and refers to a group:

[0219] [ka]

[0220] [In the formula, R 90 and R 100 independently represent hydrogen or a hydrocarbyl group such as an alkyl group, or R 90 and R 100 together with the intervening atoms complete a heterocyclic ring having 4 to 8 atoms in the ring structure. The terms "carbocycle" and "carbocyclic" as used herein refer to a saturated or unsaturated ring in which each atom of the ring is carbon. The term carbocycle includes both aromatic and non-aromatic carbocycles. Non-aromatic carbocycles include both cycloalkane rings in which all carbon atoms are saturated, and cycloalkene rings that contain at least one double bond. "Carbocycle" includes 5-7 membered monocyclic rings and 8-12 membered bicyclic rings. Each ring of a bicyclic carbocycle can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. Carbocycles include bicyclic molecules in which one, two, or three or more atoms are shared between the two rings. The term "fused carbocycle" refers to a bicyclic carbocycle in which each of the rings shares two adjacent atoms with the other ring. Each ring of a fused carbocycle can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. In exemplary embodiments, an aromatic ring (e.g., phenyl) can be fused to a saturated or unsaturated ring (e.g., cyclohexane, cyclopentane, or cyclohexene). Any combination of saturated, unsaturated, and aromatic bicyclic rings is included in the definition of carbocycle, where valence permits. Exemplary "carbocycles" include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene, and adamantane. Exemplary fused carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hept-3-ene. A "carbocycle" may be optionally substituted at any one or more positions capable of carrying a hydrogen atom.

[0221] A "cycloalkyl" group is a cyclic hydrocarbon that is fully saturated. "Cycloalkyl" includes monocyclic and bicyclic rings. Typically, a monocyclic cycloalkyl group has 3 to about 10 carbon atoms, more typically 3 to 8 carbon atoms, unless otherwise specified. The second ring of a bicyclic cycloalkyl can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. Cycloalkyl includes bicyclic molecules in which one, two, or three or more atoms are shared between the two rings. The term "fused cycloalkyl" refers to a bicyclic cycloalkyl in which each of the rings shares two adjacent atoms with the other ring. The second ring of a fused bicyclic cycloalkyl can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. A "cycloalkenyl" group is a cyclic hydrocarbon that contains one or more double bonds.

[0222] The term "carbocyclylalkyl," as used herein, refers to an alkyl group substituted with a carbocycle group.

[0223] The term “carbonate” is art-recognized and refers to the group —OCO—R 10 (In the formula, R 10 represents a hydrocarbyl group).

[0224] The term "carboxy" as used herein means a group represented by the formula -CO2H.

[0225] The term "ester" as used herein refers to the group -C(O)OR 10 (In the formula, R 10 represents a hydrocarbyl group).

[0226] The term "ether" as used herein means a hydrocarbyl group linked to another hydrocarbyl group through an oxygen. Thus, an ether substituent of a hydrocarbyl group can be hydrocarbyl-O-. Ethers can be either symmetrical or asymmetrical. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include "alkoxyalkyl" groups, which can be represented by the general formula: alkyl-O-alkyl.

[0227] The terms "halo" and "halogen" as used herein mean halogens and include chloro, fluoro, bromo, and iodo.

[0228] The terms "hetaralkyl" and "heteroaralkyl," as used herein, refer to an alkyl group substituted with a hetaryl group.

[0229] The term "heteroalkyl," as used herein, means a saturated or unsaturated chain of carbon atoms and at least one heteroatom, where no two heteroatoms are adjacent.

[0230] The term "heteroalkylamino," as used herein, refers to an amino group substituted with a heteroalkyl group.

[0231] The terms "heteroaryl" and "hetaryl" include substituted or unsubstituted aromatic monocyclic structures, preferably 5-7 membered rings, more preferably 5-6 membered rings, whose ring structures contain at least one heteroatom, preferably 1-4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more cyclic rings, where two or more carbons are common to the two connected rings, where at least one of the rings is heteroaromatic and the other cyclic ring can be, for example, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine.

[0232] The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0233] The terms "heterocyclyl", "heterocycle", and "heterocyclic" refer to a substituted or unsubstituted non-aromatic ring structure, preferably 3-10 membered, more preferably 3-7 membered, whose ring structure contains at least one heteroatom, preferably 1-4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two connected rings, where at least one of the rings is heterocyclic and, for example, the other cyclic ring can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like. Heterocyclyl groups may also be substituted with an oxo group. For example, "heterocyclyl" includes both pyrrolidine and pyrrolidinone.

[0234] The term "heterocycloalkyl," as used herein, refers to an alkyl group substituted with a heterocycle group.

[0235] The term "heterocycloalkylamino," as used herein, refers to an amino group substituted with a heterocycloalkyl group.

[0236] The term "hydrocarbyl" as used herein means a group that does not have =O or =S substituents and typically has at least one carbon-hydrogen bond and a backbone that is primarily carbon, but may optionally contain heteroatoms, and is bonded through carbon atoms. That is, groups such as methyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered hydrocarbyl for the purposes of this application, while substituents such as acetyl (which has =O substituent on the bonding carbon) and ethoxy (which is linked through oxygen rather than carbon) are not considered hydrocarbyl. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocycle, heterocyclyl, alkyl, alkenyl, alkynyl, and combinations thereof.

[0237] The term "hydroxyalkyl," as used herein, refers to an alkyl group substituted with a hydroxy group.

[0238] The term "lower" when used in connection with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups with 10 or fewer non-hydrogen atoms, preferably 6 or fewer non-hydrogen atoms in the substituent. "Lower alkyl" means, for example, an alkyl group containing 10 or fewer carbon atoms, preferably 6 or fewer carbon atoms. In certain embodiments, the acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents defined herein are lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, respectively, whether they are present alone or in combination with other substituents (such as in the description of hydroxyalkyl and aralkyl (where, for example, atoms in an aryl group are not counted when counting the carbon atoms in an alkyl substituent)).

[0239] As used herein, the term "oxo" refers to a carbonyl group. When an oxo substituent appears on an otherwise saturated group, such as an oxo-substituted cycloalkyl group (e.g., 3-oxo-cyclobutyl), the substituted group is still intended to be a saturated group. When a group is described as being substituted with an "oxo" group, this may mean that a carbonyl moiety (i.e., -C(=O)-) is replaced with a methylene unit (i.e., -CH2-).

[0240] The terms "polycyclyl," "polycycle," and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl) in which two or more atoms are common to two connected rings (e.g., the rings are "fused rings"). Each of the rings of a polycycle can be substituted or unsubstituted. In certain embodiments, each ring of a polycycle contains from 3 to 10 atoms in the ring, preferably from 5 to 7 atoms.

[0241] The term "silyl" means a silicon moiety having three hydrocarbyl moieties attached thereto.

[0242] The term "substituted" refers to a moiety having a substituent replacing a hydrogen on one or more carbons of the backbone. It will be understood that "substituted" or "substituted with" includes the implicit proviso that such substitution is in accordance with the permissible valencies of the atom and substituent being substituted, as well as that the substitution results in a stable compound (e.g., one that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, and the like). As used herein, the term "substituted" is considered to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, heteroatoms such as nitrogen can have hydrogen substituents and / or any permissible substituent of organic compounds described herein that satisfies the valence of the heteroatom. Substituents include any of the substituents described herein, such as alkyl, alkylamino, aminoalkyl, alkylaminoalkyl, halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamide, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. Those skilled in the art will understand that the substituents may themselves be substituted, where appropriate. Unless specifically described as "unsubstituted," references to chemical moieties herein are understood to include substituted variations. For example, references to an "aryl" group or moiety implicitly include both substituted and unsubstituted variations.

[0243] The term "sulfate" is art recognized and refers to the group -OSO3H, or a pharma- ceutically acceptable salt thereof.

[0244] The term "sulfonamide" is art recognized and has the general formula:

[0245] [ka]

[0246] [In the formula, R 90 and R 100 independently represents hydrogen or a hydrocarbyl such as alkyl, or R 90 and R 100 together with the intervening atoms complete a heterocyclic ring having 4 to 8 atoms in the ring structure. It means a group represented by:

[0247] The term "sulfoxide" is art recognized and refers to the group -S(O)-R, where R represents hydrocarbyl.

[0248] The term "sulfonate" is art recognized and refers to the group SO3H, or a pharma- ceutically acceptable salt thereof.

[0249] The term "sulfone" is art recognized and refers to the group -S(O)2-R, where R represents hydrocarbyl.

[0250] The term "thioalkyl," as used herein, refers to an alkyl group substituted with a thiol group.

[0251] The term "thioester," as used herein, refers to the group -C(O)SR or -SC(O)R, where R 10 represents hydrocarbyl).

[0252] The term "thioether" as used herein is equivalent to an ether where the oxygen has been replaced with a sulfur.

[0253] The term "urea" is art recognized and has the general formula:

[0254] [ka]

[0255] [In the formula, R 90 and R 100 independently represents hydrogen or a hydrocarbyl such as alkyl, or R 90 and R 100 any occurrence of together with the intervening atoms completes a heterocycle having 4 to 8 atoms in the ring structure. It can be expressed as follows.

[0256] "Protective group" refers to a group of atoms that, when attached to a reactive functional group in a molecule, masks, reduces or prevents the reactivity of the functional group. Typically, a protective group can be selectively removed, if desired, during the synthetic process. Examples of protective groups can be found in Greene and Wuts, Protective Groups in Organic Chemistry, 3rd Ed., 1999, John Wiley & Sons, NY and Harrison et al., Compendium of Synthetic Organic Methods, Vols. 1-8, 1971-1996, John Wiley & Sons, NY. Representative nitrogen protecting groups include, but are not limited to, formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl ("CBZ"), tert-butoxycarbonyl ("Boc"), trimethylsilyl ("TMS"), 2-trimethylsilyl-ethanesulfonyl ("TES"), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl ("FMOC"), nitroveratryloxycarbonyl ("NVOC"), etc. Representative hydroxyl protecting groups include, but are not limited to, those in which the hydroxyl group is acylated (esterified) or alkylated, such as benzyl and trityl ethers, as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers (e.g., TMS or TIPS groups), glycol ethers (such as ethylene glycol and propylene glycol derivatives), and allyl ethers.

[0257] As used herein, a therapeutic agent that "prevents" a disorder or condition refers to a compound that reduces the occurrence of the disorder or condition in a statistical sample in a treated sample compared to an untreated control sample, or delays the onset of the disorder or condition or reduces the severity of one or more symptoms compared to an untreated control sample.

[0258] The term "treating" includes prophylactic and / or therapeutic treatment. The term "prophylactic or therapeutic" treatment is art-recognized and includes administration of one or more of the subject compositions to a host. When administered prior to the clinical manifestation of an undesirable condition (e.g., a disease or other undesirable condition of a host animal), the treatment is prophylactic (i.e., protects the host from developing the undesirable condition), whereas when administered after the manifestation of the undesirable condition, the treatment is therapeutic (i.e., is intended to eliminate, ameliorate, or stabilize an existing undesirable condition or its side effects).

[0259] The term "prodrug" is intended to encompass compounds that are converted under physiological conditions into the therapeutically effective agents of the present invention (e.g., compounds of formula I). ​​A common method for making prodrugs is to include one or more selected moieties that are hydrolyzed under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by the enzyme activity of the host animal. For example, esters or carbonates (e.g., esters or carbonates of alcohols or carboxylic acids) are preferred prodrugs of the present invention. In certain embodiments, some or all of the compounds of formula I in the formulations presented above can be replaced with the corresponding suitable prodrugs, for example, a hydroxyl in the parent compound is presented as an ester or carbonate, or a carboxylic acid present in the parent compound is presented as an ester. The term "MK2-associated disorder" refers to a disorder or condition in which MK2 plays a role in the pathology or symptoms of a disease or disorder. For example, disorders associated with MK2 include, but are not limited to, inflammatory diseases, autoimmune diseases, destructive bone disorders, cell proliferative disorders, angiogenic disorders, infectious diseases, neurodegenerative diseases, and viral diseases.

[0260] As used herein, the term "inflammatory disorder" or "inflammatory disease" includes diseases and disorders that are caused or primarily caused by inflammation, as well as diseases and disorders in which inflammation plays a role in the pathology or symptoms of the disease or disorder, the spread of the disease or disorder, the worsening of the symptoms of the disease or disorder, and / or the worsening of the patient prognosis or survival due to the disease or disorder.

[0261] [Example]

[0262] A. Chemical synthesis The general procedures used in the methods of preparing the compounds of this invention are described below.

[0263] General method All solvents and reagents were obtained from commercial sources and used without further purification unless otherwise indicated. NMR spectra were obtained on a Bruker Neo 400M spectrometer operating at 400 MHz. Chemical shifts are reported in parts per million (δ) from the tetramethylsilane resonance in the indicated solvent. LC-mass spectra were obtained using an Agilent 1260-6125B single quadrupole mass spectrometer, using a Welch Biomate column (C18, 2.7 um, 4.6×50 mm) and eluting with a mixture of solvents A (ACN with 0.05% TFA) and B (water with 0.05% TFA) using gradient elution. Detection was by DAD (254 nm and 210 nm). Ionization was by ESI. Spectra were analyzed using Chemstation software. Analytical HPLC was performed using gradient elution on a Waters ARC system, either under acidic conditions on a YMC Pack Pro column (C18 S-3um, 12nm, 150x2.0mm) eluting with a mixture of solvents A (ACN with 0.05% TFA) and B (water with 0.05% TFA); or under base-containing conditions on an Agilent Poroshell HPH C18 column (2.7um, 2.1x150mm) eluting with a mixture of solvents C (water with 0.1% NH4OH) and D (ACN with 0.1% NH4OH). Detection was by DAD (254nm and 210nm). Preparative HPLC was performed using a Welch C18 column (5um, 25x150mm) eluting with a mixture of solvents A and B, on a Waters AutoP system coupled with a single quadrupole mass spectrometer. Flash chromatography was performed using a Biotage Isolera Prime system using Welch WelFlash flash columns (40-63 um) eluted with mixtures of solvents as indicated in the experimental procedures.

[0264] [ka]

[0265] Methyl 4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (Int-A-1) A suspension of methyl 4-bromo-1H-pyrrole-2-carboxylate (100 g, 489 mmol), ((4-cyanophenyl)boronic acid (108 g, 734.6 mmol), K2CO3 (202 g, 1467 mmol, 3.0 equiv), Pd(dppf)Cl2 (17.9 g, 24.5 mmol, 0.05 equiv)) in dioxane / H2O (1.8 L, 5:1) was degassed and then heated to 100 °C for 20 h under N2. The reaction mixture was then cooled to room temperature and poured into ice water (2 L) with stirring. The formed precipitate was collected by filtration, which was redissolved in DCM (ca. 3 L) and washed with brine. The organic solvent was passed through a short silica gel column and eluted with PE:DCM (1:1). The product-containing fractions were combined and concentrated under reduced pressure. The residue was redissolved in DCM (approximately 1 L) and to this was slowly added 10 L of petroleum ether to precipitate the product. After stirring for 1 h, the solid was collected by filtration and dried to give the desired product (92 g, 93% purity) as a pale yellow solid. ESI: [M+H] + = 227.1; 1 H NMR (400MHz, DMSO-d6): δ12.30 (br. s., 1H), 7.84 (d, J =8.4 Hz, 2H), 7.79 - 7.69 (m, 3H), 7.33 (br. s., 1H), 3.80 (s, 3H).

[0266] (5-Bromo-2-nitrophenyl)methanol (Int-A-2) To a solution of 5-bromo-2-nitrobenzaldehyde (250 g, 1.086 mol) in MeOH (1.0 L) at 0° C. was added NaBH4 (12.4 g, 326 mmol) batchwise over 30 min. After the addition, the mixture was allowed to warm to room temperature and continued to stir for 10 min. The mixture was then quenched by slow addition of saturated NH4Cl (1 L) and concentrated under reduced pressure to remove volatile solvents. The residue was extracted with EA (3×1 L) and the combined organic layers were washed with brine, dried over Na2SO4, then concentrated under reduced pressure to give the desired product (252 g, 1086 mmol) as a white solid. ESI: [M+H] + = 232.2 / 234.2; 1 H NMR (400MHz, CDCl3): δ 7.98 - 7.91 (m, 2H), 7.55 (dd, J =1.8, 8.8 Hz, 1H), 4.97 (br. s., 2H), 2.77 (br. s, 1H).

[0267] Methyl 1-(5-bromo-2-nitrobenzyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (Int-A-3) To a solution of compound Int-A-2 (105 g, 442.5 mmol) and TEA (123 mL, 885 mmol, 2.0 equiv.) in DCM (1.5 L), MsCl (37.6 mL, 486.7 mmol, 1.1 equiv.) was added at 0° C., and the mixture was stirred at 20° C. for 0.5 h. Compound Int-A-1 (100 g, 442.5 mmol) was added to the solution, followed by a solution of tetrabutylammonium hydroxide (45 g, 44.2 mmol, 0.1 equiv.) and NaOH (2 M, 1.1 L, 5.0 equiv.) at 0° C. Then, the mixture was stirred at room temperature for 2 h. The resulting yellow solid was collected by filtration. To the residue, EtOH (200 mL) was added and stirred for 1 h. The precipitate was collected by filtration to give the desired product (280 g, 636 mmol, 71.9% yield) as an off-white solid. ESI: [M+H] + = 440.1 / 442.1; 1H NMR (400MHz, DMSO-d6): δ 8.08 (d, J = 8.8 Hz, 1H), 8.00 (d, J = 1.8 Hz, 1H), 7.84 - 7.76 (m, 5H), 7.55 (d, J = 1.8 Hz, 1H), 6.71 (d, J = 1.3 Hz, 1H), 5.86 (s, 2H), 3.29 (s, 3H).

[0268] Methyl 1-(2-amino-5-bromobenzyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (Int-A-4) A suspension of compound Int-A-3 (186 g, 423.4 mmol), Fe (118.2 g, 2.1 mol) and NH4Cl (226.5 g, 4.2 mol, 10.0 equiv) in EtOH (2 L), H2O (1 L) and THF (2 L) was heated to 80° C. for 2 h. The mixture was then concentrated to dryness and the residue was redissolved in hot THF (20 L*5). The mixture was filtered through a celite pad and the filtrate was concentrated. The residue was slurried with EtOH (1 L) and stirred for 1 h. The solid was collected by filtration to give the desired product (138 g). ESI: [M+H] + = 410.2 / 412.2; 1 H NMR (400MHz, DMSO-d6): δ 7.88 (br. s., 1H), 7.85 - 7.68 (m, 4H), 7.50 (br. s., 1H), 7.09 (d, J = 7.9 Hz, 1H), 6.62 (d, J = 8.4 Hz, 1H), 6.33 (br. s., 1H), 5.37 (br. s., 4H), 3.73 (br. s., 3H).

[0269] 4-(7-Bromo-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (Int-A-5) To a suspension of compound Int-A-4 (100 g, 244 mmol) in toluene (1 L), Me3Al (2M in toluene, 609 mL) was added dropwise at 0° C., and the mixture was stirred at 20° C. for 10 h under N2. The mixture was then poured into 1M ice-cold HCl (2M, 500 mL). The resulting mixture was filtered, and the filter cake was washed with NaHCO3 (1 L). The solid was slurried in EtOH (700 mL), stirred for 2 h, and then filtered off. After drying under vacuum, the desired product (86.2 g, 166 mmol, 93.7% yield) was obtained as a yellow solid. ESI: [M+H] + = 378.2 / 380.2; 1 H NMR (400MHz, DMSO-d6): δ 10.30 (s, 1H), 7.73 (s, 4H), 7.66 (dd, J = 2.0, 4.2 Hz, 2H), 7.50 (dd, J = 2.2, 8.4 Hz, 1H), 7.27 (d, J = 1.8 Hz, 1H), 7.13 (d, J = 8.8 Hz, 1H), 5.21 (s, 2H).

[0270] 4-(7-Bromo-11-chloro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (Int-A-6) A solution of compound Int-A-5 (80 g, 211 mol) in POCl3 (300 mL) was stirred at 90° C. for 18 h. The solution was concentrated under reduced pressure and the residue was dissolved in EA (3 L). The organic solution was poured into ice-cold NaHCO3 solution (2 L) and stirred for 30 min. The organic layer was separated, washed with brine and dried over Na2SO4. After concentration under reduced pressure, the crude product (76 g, 93.7% yield) was used immediately in the next step. ESI: [M+H] + = 396.2 / 398.2

[0271] 4-(7-Bromo-11-((2,2-dimethoxyethyl)amino)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (Int-A-7) To a solution of compound Int-A-6 (75 g, 190 mmol) in THF / dioxane (500 mL / 1 L) was added 2,2-dimethoxyethanamine (100 g, 949 mmol). The resulting solution was stirred at 100° C. for 18 h. Then the mixture was concentrated under reduced pressure. The residue was redissolved in EA (300 mL) and HCl (1 M, 1.2 L) was added slowly to the above solution. A precipitate formed during the addition, which was filtered off. The filter cake was washed several times with EA and dried under vacuum to give the desired product (wet solid), which was used in the next step without further purification. ESI: [M+H] + = 465.2 / 467.2

[0272] 4-(7-Bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (Int-A) To a solution of the above solid in dioxane (800 mL), HCl (1M, 400 mL) was added and the resulting mixture was stirred at 80° C. for 20 h. Then, the reaction mixture was cooled to room temperature and filtered. The filter cake was washed with saturated NaHCO3 solution (1 L) followed by deionized water (1 L). Then, the solid was reslurried in MeOH (300 mL). After filtration, the filter cake was collected and dried under vacuum to give the desired product as a white solid (36 g). ESI: [M+H] + = 401.2 / 403.2; 1 H NMR (400 MHz, DMSO-d6): δ 7.92 (d, J = 2.3 Hz, 1H), 7.84 (d, J = 1.4 Hz, 1H), 7.75-7.774(m, 4H), 7.65 (d, J = 1.9 Hz, 1H), 7.56 (d, J = 8.6 Hz, 1H), 7.25 (d, J = 1.4 Hz, 1H), 7.06 (d, J = 1.9 Hz, 1H), 5.24 (s, 2H).

[0273] [ka]

[0274] (S)-3,4-Dihydroxybutanamide (1-A) A solution of (S)-4-hydroxydihydrofuran-2(3H)-one (5 g, 49 mmol) in NH3 solution in MeOH (7 mol / L, 12 mL) was stirred at 25° C. for 19 h. The mixture was concentrated to give the desired product (6 g, 100% yield). ESI: [M+H] + =120.1

[0275] (S)-3-Hydroxy-4-(trityloxy)butanamide (1-B) To a solution of compound 011-A (6 g, 50.39 mmol) and pyridine (6 mL) in THF (25 mL) and DMF (25 mL), trityl chloride (15.45 g, 55.43 mmol) was added and stirred at 25° C. for 18 h. Then, to the mixture, NH4Cl aqueous solution (200 mL) and EA (200 mL) were added. After stirring for 30 min, the organic layer in the mixture was separated, washed with brine, dried over anhydrous Na2SO4, filtered and concentrated to give the desired compound as a yellow solid (5.3 g, yield: 29%). ESI: [M+H] = 362.1

[0276] (S)-4-(((2-oxooxazolidin-5-yl)methoxy)diphenylmethyl)benzene-1-ylium (1-C) To a solution of compound 1-B (5.67 g, 15.7 mmol) and 13% NaClO (25.2 mL) in THF (47 mL) was added NaOH (2.52 g 63.5 mmol) in H2O (15.75 mL). The resulting mixture was stirred at 60 °C for 1 h before the addition of water (200 mL) and EA (200 mL). The organic layer in the mixture was separated, washed with brine and dried over anhydrous Na2SO4. After filtration and concentration under reduced pressure, the residue was purified by chromatography on silica gel (DCM:MeOH = 40:1) to give the desired compound (3.87 g, yield: 68.6%). ESI: [M+H] + = 360.2

[0277] General Procedure I - Ullmann Reaction: (S)-4-(7-(2-oxo-5-((trityloxy)methyl)oxazolidin-3-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (1-D) A solution of Int-A (100 mg, 0.25 mmol), compound 1-C (134.25 mg, 0.37 mmol), CuI (4.75 mg, 0.025 mmol), Cs2CO3 (162.4 mg, 0.5 mmol), trans-N,N'-dimethylcyclohexane-1,2-diamine (7.09 mg, 0.05 mmol) in anhydrous DMSO (2 mL) was stirred under N2 at 115 °C for 16 h. The reaction mixture was then cooled to room temperature and H2O (10 mL), EA (10 mL), and NH4OH:NH4Cl = 1:10 (30 mL) were added. The resulting mixture was stirred and then filtered. The organic layer in the filtrate was separated, washed with brine, and dried over anhydrous Na2SO4. After filtration and concentration, the residue was purified by chromatography on silica gel (DCM:MeOH=30:1) to give the desired compound as a brown solid (230 mg, yield >99%). ESI: [M+H] + = 680.3

[0278] (S)-4-(7-(5-(hydroxymethyl)-2-oxooxazolidin-3-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (1-E) To a solution of compound 1-D (230 mg, 0.34 mmol) in DCM (10 mL) was added TFA (0.5 mL). After stirring at 25° C. for 30 min, NaHCO (20 mL) and DCM (20 mL) were added to the solution. The organic layer was separated, washed with brine, and dried over anhydrous NaSO. After filtration and concentration, the residue was purified by chromatography on silica gel (DCM:MeOH=100:1) to give the desired product as a brown solid (117.4 mg, yield: 73%). ESI: [M+H] + = 348.1

[0279] General Procedure II: (R)-4-(7-(5-((dimethylamino)methyl)-2-oxooxazolidin-3-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (1) To a solution of compound 1-E (100 mg, 0.23 mmol), TEA (46.25 mg, 0.46 mmol) in DCM (50 mL) was added MsCl (0.1 mL, 1.26 mmol) at 0° C., and the resulting solution was stirred at 0° C. for 3.5 h. The mixture was diluted with ice water (100 mL) and extracted with DCM (100 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, and filtered. After filtration and concentration, the crude compound was obtained as a brown solid (115 mg), which was used directly in the next step. ESI: [M+H] + = 516.1. A solution of the above compound (50 mg, 0.1 mmol) in DMF (2 mL), Me2NH (2 mL, 2 mmol) in THF was placed in a microwave reactor at 100 °C for 2 h. The mixture was diluted with H2O (50 mL) and extracted with DCM (50 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated. The resulting residue was purified by preparative HPLC to give the desired compound as a white solid (11.1 mg, yield: 23%). ESI: [M+H] + = 465.2; 1 H-NMR (400 MHz, DMSO-d6): δ 9.9 (brs,1H), 7.6-8.0 (m, 10H), 7.55 (s, 1H), 5.36 (s, 2H), 5.20 (s, 1H), 4.30 (t, 1H), 3.88 (s, 2H), 2.89 (s, 6H), 2.68 (s, 1H), 2.38 (s, 1H).

[0280] Using the procedures described above, the examples in Table 3 below were synthesized.

[0281] [Table 3]

[0282] [ka]

[0283] (R)-4-(7-(5-(((4-hydroxy-2-methylbutan-2-yl)amino)methyl)-2-oxooxazolidin-3-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (6-A) Compound 6-A was synthesized using the procedure described above. ESI: [M+H] + = 523.2.

[0284] (R)-4-(7-(5-((2,2-dimethylazetidin-1-yl)methyl)-2-oxooxazolidin-3-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (6) To a solution of compound 6-A in dry DCM (1 mL), SOCl2 (1 drop) was added and the resulting solution was stirred at 40°C for 30 min. After the reaction was completed, the mixture was concentrated to give the crude compound (11 mg). ESI: [M+H] + = 541.2. To a solution of the above compound (11 mg, 0.02 mmol) in dry DMF (1 mL), K2CO3 (10 mg) was added and the mixture was placed in a microwave reactor at 80 °C for 2 h. The reaction mixture was concentrated and purified by preparative HPLC to give the desired compound (2 mg, yield: 20%). ESI: [M+H] + = 505.2; 1H NMR (400 MHz, MeOH-d4): δ 8.01 (dd, J = 10.2, 2.2 Hz, 2H), 7.80 (dd, J = 8.9, 2.5 Hz, 1H), 7.78 - 7.73 (m, 4H), 7.72 - 7.68 (m, 2H), 7.65 (d, J = 1.8 Hz, 1H), 7.27 (d, J = 1.8 Hz, 1H), 5.37 (s, 2H), 5.03 (s, 1H), 4.36 (t, J = 9.1 Hz, 1H), 4.16 (s, 3H), 3.90 (dd, J = 9.2, 6.4 Hz, 1H), 3.59 (s, 2H), 2.57 (s, 1H), 2.29 (s, 1H), 1.69 (d, J = 6.3 Hz, 6H).

[0285] [ka]

[0286] 1-((S)-1-(4-Methoxyphenyl)ethyl)-5-oxopyrrolidine-3-carboxylic acid (7-A) (S)-1-(4-Methoxyphenyl)ethanamine in 1-methyl-2-pyrrolidinone (80 mL) A suspension of (13.5 g, 99.15 mmol) and itaconic acid (13 g, 98.2 mmol) was heated to 80° C. for 1 h. The solution was stirred at 120° C. for an additional 4 h. The reaction mixture was cooled to 25° C. and poured into 500 mL of demineralized water. The precipitate formed was filtered, washed with demineralized water and dried at 50° C. to give the desired compound as a white solid (21 g, 79.8 mmol, 89% yield).

[0287] (S)-Benzyl-1-((S)-1-(4-methoxyphenyl)ethyl)-5-oxopyrrolidine-3-carboxylate (7-B). A solution of compound 7-A (21 g, 79.8 mmol), benzyl bromide (10.34 mL, 87.8 mmol, 1.1 equiv.) and cesium carbonate (28.55 g, 87.8 mmol, 1.1 equiv.) in DMF (100 mL) was stirred at room temperature for 45 min. Water and ethyl acetate were added to the reaction mixture. The organic layer was separated and the aqueous layer was extracted with EtOAc (200 mL*2). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (solvent gradient: 5 percent to 50 percent petroleum ether / ethyl acetate) to give (S)-benzyl 1-((S)-1-(4-methoxyphenyl)ethyl)-5-oxopyrrolidine-3-carboxylate (11.2 g, 31.73 mmol, 39.7% yield) as the first eluate and (R)-benzyl 1-((S)-1-(4-methoxyphenyl)ethyl)-5-oxopyrrolidine-3-carboxylate (11.6 g, 32.86 mmol, 41.2% yield) as the second eluate. 7-B: 1 H NMR (400 MHz, CDCl3): δ 7.42 - 7.28 (m, 5H), 7.20 (t, J = 5.7 Hz, 2H), 6.88 - 6.82 (m, 2H), 5.44 (q, J = 7.1 Hz, 1H), 5.14 (s, 2H), 3.79 (s, 3H), 3.53 (dd, J = 9.4, 5.8 Hz, 1H), 3.22 - 3.05 (m, 2H), 2.73 (qd, J = 17.1, 8.4 Hz, 2H), 1.48 (d, J = 7.1 Hz, 3H) 7-B': 1H NMR (400 MHz, CDCl3): δ 7.38 - 7.30 (m, 3H), 7.27 - 7.22 (m, 2H), 7.18 (t, J = 5.7 Hz, 2H), 6.88 - 6.80 (m, 2H), 5.44 (q, J = 7.1 Hz, 1H), 5.06 (d, J = 1.5 Hz, 2H), 3.77 (d, J = 9.0 Hz, 3H), 3.58 - 3.47 (m, 1H), 3.28 - 3.14 (m, 2H), 2.80 - 2.61 (m, 2H), 1.49 (d, J = 7.1 Hz, 3H). ESI: [M+H] + = 354

[0288] (S)-4-(hydroxymethyl)-1-((S)-1-(4-methoxyphenyl)ethyl)pyrrolidin-2-one (007-C) To a solution of compound 7-B (11.2 g, 31.7 mmol) in EtOH (75 mL) was added NaBH4 (4.2 g, 111 mmol) at 0 °C. The mixture was stirred at room temperature for 8 h. The reaction was quenched by the addition of NH4Cl. EtOAc was added and the organic layer was separated. The aqueous layer was extracted with EtOAc (3*200 mL). The combined organic layers were washed with saturated salt, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (PE:EA=5:1 to DCM:MeOH=100:1) to give the desired alcohol (7.5 g, 30.12 mmol, 95% yield). ESI: [M+H] + =250

[0289] (S)-4-(hydroxymethyl)pyrrolidin-2-one-D) A solution of compound 7-C (7.5 g, 30.12 mmol, 1.0 equiv) in TFA (40 mL) was heated to 80 °C and stirred for 16 h. The volatile solvents were removed under reduced pressure. The residue was redissolved in MeOH (50 mL) and NH3·H2O was added to the above solution at room temperature. After stirring for 1 h, the reaction mixture was concentrated and the residue was purified by silica gel column chromatography (DCM:MeOH=100:1 to 10:1) to give the desired product (3.0 g, 26.1 mmol). ESI: [M+H] + = 116; 1 H NMR (400 MHz, MeOH-d4): δ 3.58 - 3.51 (m, 2H), 3.51 - 3.45 (m, 1H), 3.20 (dd, J = 10.1, 5.3 Hz, 1H), 2.72 - 2.56 (m, 1H), 2.42 (dd, J = 17.1, 9.1 Hz, 1H), 2.24 - 2.05 (m, 1H).

[0290] (S)-4-(7-(4-(hydroxymethyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (07-E) Compound 7-E was synthesized using the procedure described above. ESI: [M+H] + = 436

[0291] (R)-4-(7-(4-((dimethylamino)methyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (7) Using the procedure described above, compound 7 was synthesized. ESI: [M+H] + = 436.3; 1H NMR (400 MHz, MeOH-d4): δ 7.97 (d, J = 2.5 Hz, 1H), 7.78 (dd, J = 8.8, 2.5 Hz, 1H), 7.74 - 7.67 (m, 3H), 7.66 - 7.62 (m, 2H), 7.58 (d, J = 8.8 Hz, 1H), 7.53 (d, J = 1.9 Hz, 1H), 7.25 (d, J = 1.3 Hz, 1H), 7.05 (d, J = 1.9 Hz, 1H), 5.18 (s, 2H), 4.05 (dd, J = 9.7, 7.8 Hz, 1H), 3.76 - 3.67 (m, 1H), 2.83 - 2.71 (m, 2H), 2.51 (dd, J = 12.3, 7.7 Hz, 1H), 2.47 - 2.37 (m, 2H), 2.30 (s, 6H).

[0292] Using the procedures described above, the examples in Table 4 below were synthesized.

[0293] [Table 4] TIFF2024504927000089.tif238170TIFF2024504927000090.tif69170

[0294] [ka]

[0295] (S)-4-(7-(4-((dimethylamino)methyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (17) Using the procedure described above, compound 17 was synthesized: ESI: [M+H] + = 463.2; 1H NMR (400 MHz, MeOH-d4): δ 8.10 (dd, J = 15.3, 2.3 Hz, 2H), 7.87 (dd, J = 8.9, 2.5 Hz, 1H), 7.80 (d, J = 1.8 Hz, 1H), 7.78 (s, 1H), 7.77 - 7.73 (m, 4H), 7.71 (d, J = 2.1 Hz, 2H), 7.70 (d, J = 2.0 Hz, 1H), 7.33 (d, J = 1.8 Hz, 1H), 5.40 (s, 2H), 4.19 (dd, J = 9.7, 8.1 Hz, 1H), 3.80 (dd, J = 9.8, 7.3 Hz, 1H), 3.41 (dd, J = 7.3, 1.1 Hz, 2H), 3.14 - 3.06 (m, 1H), 2.99 (d, J = 3.3 Hz, 6H), 2.91 (dd, J = 16.9, 8.7 Hz, 1H), 2.58 (dd, J = 17.0, 8.5 Hz, 1H).

[0296] [ka]

[0297] (S)-1-(1-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-5-oxopyrrolidin-3-yl)-N,N,N-trimethylmethanaminium iodide (18) To a solution of compound 007 (10 mg, 0.02 mmol) in DMF (2 mL) was added iodomethane (0.5 mL) at room temperature. The mixture was stirred at 20° C. for 2 h. Then, the solvent was removed in vacuo. The residue was purified by preparative HPLC to give the desired product (1 mg, yield: 8%). ESI: [M+H] + = 477.2; 11H NMR (400 MHz, DMSO-d6): δ 8.04 (d, J = 2.2 Hz, 1H), 7.96 (s, 1H), 7.76 (dd, J = 16.3, 8.6 Hz, 6H), 7.67 (d, J = 8.8 Hz, 1H), 7.45 (s, 1H), 7.15 (s, 1H), 5.31 (s, 2H), 4.15 (t, J = 9.0 Hz, 1H), 3.72 (t, J = 9.1 Hz, 1H), 3.57 (d, J = 5.9 Hz, 2H), 3.13 (s, 9H), 2.85 (dd, J = 16.6, 8.7 Hz, 1H), 2.69 - 2.65 (m, 1H), 2.34 - 2.31 (m, 2H).

[0298] [Chemical formula]

[0299] Compound 19 was synthesized using the above procedure. ESI: [M+H] + = 503.2; 1H NMR (400 MHz, MeOH-d4): δ 7.97 (d, J = 2.4 Hz, 1H), 7.77 (dd, J = 8.8, 2.5 Hz, 1H), 7.74 - 7.67 (m, 3H), 7.67 - 7.62 (m, 2H), 7.58 (d, J = 8.8 Hz, 1H), 7.54 (d, J = 1.9 Hz, 1H), 7.25 (d, J = 1.4 Hz, 1H), 7.05 (d, J = 1.9 Hz, 1H), 5.19 (s, 2H), 4.58 (s, 1H), 4.03 (dd, J = 9.7, 7.4 Hz, 1H), 3.70 (dd, J = 9.8, 5.9 Hz, 1H), 2.74 (dd, J = 16.9, 8.1 Hz, 1H), 2.56 (dd, J = 16.7, 9.7 Hz, 3H), 2.41 (dd, J = 16.9, 6.9 Hz, 1H), 1.93 (t, J = 7.0 Hz, 2H), 1.25 (d, J = 2.5 Hz, 6H).

[0300] [ka]

[0301] tert-Butyl (1-hydroxy-2-methylpropan-2-yl)carbamate (20-A) To a solution of 2-amino-2-methylpropan-1-ol (44.5 g, 0.5 mol) and TEA (60.7 g, 0.6 mol) in DCM (200 mL) was added Boc2O (109.1 g, 0.5 mol) in portions at 0° C., and then the mixture was stirred at 20° C. for 16 h. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography eluting with PE / EA (10 / 1 to 3 / 1) to give the desired product (94.6 g, yield: 94%). ESI: [M+H] + = 190

[0302] tert-Butyl (2-methyl-1-oxopropan-2-yl)carbamate (20-B) To a solution of compound 20-A (20 g, 105.7 mmol) in DCM (200 mL) was added DMP (53.8 g, 126.8 mmol) slowly at 0° C., and then the mixture was stirred at 20° C. for 3 h. The mixture was then quenched with saturated aqueous NaHCO3 and Na2SO3 solutions. The mixture was then stirred for 0.5 h, extracted with DCM, washed with brine, and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography eluting with PE / EA (10 / 1 to 3 / 1) to give the desired compound (15.8 g, yield: 79.9%). ESI: [M+H] + = 188; 1 H NMR (400 MHz, CDCl3): δ 9.42 (s, 1H), 1.43 (s, 9H), 1.32 (s, 6H).

[0303] tert-Butyl (3-hydroxy-2-methyl-4-nitrobutan-2-yl)carbamate (20-C) To a solution of compound 20-B (8 g, 42.7 mmol) and nitromethane (2.6 g, 42.7 mmol) in EtOH (80 mL), NaOH (10 mol / L, 4.3 mL) was added dropwise at 0° C., and then the mixture was stirred at 20° C. for 16 h. The mixture was quenched with HOAc until pH<6 and extracted with EA. The organic phase was washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with PE / EA (10 / 1 to 1 / 1) to give the desired product (9.2 g, yield: 86.7%). ESI: [M+H] + = 249; 1 H NMR (400 MHz, CDCl3): δ 4.60 (s, 1H), 4.55 (dd, J = 12.3, 2.4 Hz, 1H), 4.38 (dd, J = 12.3, 9.5 Hz, 1H), 4.27 (d, J = 9.3 Hz, 1H), 1.44 (s, 9H), 1.42 (s, 3H), 1.29 (s, 3H).

[0304] tert-Butyl (4-amino-3-hydroxy-2-methylbutan-2-yl)carbamate (20-D) To a solution of compound 20-C (4.0 g, 16.1 mmol) in CF3CH2OH (100 mL) was added Pd / C (0.4 g, 10%) and the mixture was stirred at 50° C. for 18 h under 50 psi hydrogen atmosphere. The mixture was cooled to room temperature and filtered through a celite pad; the filtrate was concentrated under reduced pressure to give the desired product (3.2 g, yield: 91%). ESI: [M+H] + = 219.

[0305] tert-Butyl (2-(2-oxooxazolidin-5-yl)propan-2-yl)carbamate (20-E) To a solution of compound 20-D (2.2 g, 9.6 mmol) in THF (30 mL) was added CDI (4.7 g, 28.9 mmol) at 0 °C under N2, and the mixture was stirred at 70 °C for 18 h under N2. The mixture was cooled to room temperature and washed with 0.5 N HCl (100 mL) to adjust pH < 7. The resulting mixture was extracted with EA. The organic phase was washed with brine and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography eluting with DCM / MeOH (100 / 1 to 20 / 1) to give the desired product (1.6 g, yield: 60.8%). ESI: [M+H] + = 245; 1 H NMR (400 MHz, CDCl3): δ 5.41 (s, 1H), 5.00 (dd, J = 8.9, 7.4 Hz, 1H), 4.62 (s, 1H), 3.55 (dt, J = 16.2, 8.7 Hz, 2H), 1.42 (s, 9H), 1.38 (s, 3H), 1.29 (s, 3H).

[0306] General Procedure III: tert-Butyl (2-(3-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-2-oxooxazolidin-5-yl)propan-2-yl)carbamate (20-F) To a suspension of compound Int-A (2.0 g, 5 mmol), compound 20-E (1.50 g, 6 mmol) in DMA (30 mL) was added Pd2(dba)3 (0.92 g, 1.0 mmol), Xantphos (1.16 g, 2 mmol) and Cs2CO3 (3.25 g, 10 mmol) at 20° C., and the mixture was stirred at 80° C. for 24 h under N2. The reaction mixture was cooled to room temperature, diluted with DCM / MeOH (20 / 1, 200 mL) and filtered. The filtrate was concentrated in vacuo and the residue was poured into ice-cold water. The formed solid was collected by filtration and purified first by silica gel chromatography eluting with DCM / MeOH (100 / 1 to 20 / 1) and then repurified by flash chromatography eluting with DCM / EA (100 / 1 to 1 / 1) to give the desired compound (2.0 g, yield: 71.4%). ESI: [M+H] + = 565.2

[0307] tert-Butyl (2-(3-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-2-oxooxazolidin-5-yl)propan-2-yl)carbamate (20) To a solution of compound 20-F (0.12 g, 0.2 mmol) in DCM / MeOH (5 mL / 5 mL) was added HCl in dioxane (4N, 2 mL) at 0° C., and the mixture was stirred at 20° C. for 2 h. The reaction mixture was concentrated in vacuo to give the desired compound as an HCl salt. The product was then purified by preparative HPLC to give the desired product as a white solid (50 mg, yield: 54%). ESI: [M+H] + = 465.2; 1H-NMR (400 MHz, CD3OD): δ 8.09 (s, 1H), 8.05 (d, J = 2.5 Hz, 1H), 7.88 (dd, J = 9.0, 2.5 Hz, 1H), 7.83 - 7.72 (m, 5H), 7.70 (d, J = 8.5 Hz, 2H), 7.34 (s, 1H), 5.42 (s, 2H), 4.84 - 4.67 (m, 1H), 4.35 (t, J = 9.6 Hz, 1H), 4.12 (dd, J = 9.8, 6.9 Hz, 1H), 1.43 (d, J = 21.4 Hz, 6H).

[0308] [ka]

[0309] (S)-tert-Butyl (2-(3-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-2-oxooxazolidin-5-yl)propan-2-yl)carbamate (21-A) Compound 20-F was separated by chiral-HPLC under the following conditions to give peak 1 (2.12 g, ee: 99.7%) and peak 2 (2.2 g, ee: 98.5%): Column: IG-H Column size: 0.46cm ID*15cmL Mobile phase: CO2:EtOH(0.1%DEA)=60:40 Flow rate: 2.5mL Wavelength: UV254nm Temperature: 25℃ Peak 1 was arbitrarily assigned as the S-configuration.

[0310] (S)-4-(7-(5-(2-aminopropan-2-yl)-2-oxooxazolidin-3-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (21) To a solution of compound 021-A (2.12 g, 3.75 mmol) in DCM / MeOH (20 mL / 30 mL) was added HCl in dioxane (4N, 20 mL) at 0° C., and the mixture was stirred at 20° C. for 8 h. The reaction mixture was concentrated in vacuo to give the desired compound as an HCl salt (1.89 g, purity >95%). The product was then slurried in THF / MeOH (20 mL / 20 mL) to give the desired product as a pale white solid (1.12 g, yield: 59%). ESI: [M+H] + = 465.2; 1 H NMR (400 MHz, MeOH-d4): δ 8.17 (d, J = 2.2 Hz, 1H), 8.10 (d, J = 2.5 Hz, 1H), 7.97-7.69 (m, 8H), 7.39 (d, J = 1.8 Hz, 1H), 5.49 (s, 2H), 4.37 (t, J = 9.6 Hz, 1H), 4.17 (dd, J = 9.9, 6.9 Hz, 1H), 1.47 (s, 3H), 1.41 (s, 3H).

[0311] [ka]

[0312] tert-Butyl 6-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-7-oxo-2,6-diazaspiro[3.4]octane-2-carboxylate (22-A) Compound 21-A was synthesized using the procedure described above. ESI: [M+H] + = 547.2

[0313] 4-(7-(7-oxo-2,6-diazaspiro[3.4]octan-6-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (22) To a solution of compound 21-A (100 mg, 0.18 mmol) in MeOH (5 mL) was added HCl-dioxane (4 M, 2 mL). The resulting solution was stirred at room temperature for 1 h and then concentrated to dryness. The residue was purified by preparative HPLC to give the desired product (50 mg, yield: 62.1%). ESI: [M+H] + = 447.2; 1 H-NMR (400 MHz, MeOH-d4): δ 8.11 - 8.05 (m, 2H), 7.87 (dd, J = 8.9, 2.5 Hz, 1H), 7.76-7.73(m, 5H), 7.69 (d, J = 8.5 Hz, 2H), 7.32 (s, 1H), 5.39 (s, 2H), 4.28 (t, J = 5.6 Hz, 4H), 4.21 (d, J = 11.3 Hz, 2H), 3.08 (s, 2H).

[0314] [ka]

[0315] 4-(7-(2-methyl-7-oxo-2,6-diazaspiro[3.4]octan-6-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (23) To a solution of compound 022 (50 mg, 0.11 mmol) in MeOH (5 mL), 10% Pd / C (100 mg), HCHO in H2O (2 mL) were added. The resulting mixture was stirred at room temperature overnight under hydrogen atmosphere. After the reaction was completed, the mixture was filtered through a celite pad. The filtrate was concentrated and purified by preparative HPLC to give the desired product (7 mg, yield: 13.4%). ESI: [M+H] + = 461.2; 1H NMR (400 MHz, MeOH-d4): δ 8.08 (d, J = 15.3 Hz, 2H), 7.85 (dd, J = 8.9, 2.3 Hz, 1H), 7.81 - 7.67 (m, 7H), 7.32 (s, 1H), 5.39 (s, 2H), 4.36-4.28 (m, 6H), 3.08 (s, 2H), 2.99 (s, 3H).

[0316] [ka]

[0317] tert-Butyl-6-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-5-oxo-2,6-diazaspiro[3.4]octane-2-carboxylate (24-A) Compound 24-A was synthesized using the procedure described above. ESI: [M+H] + =547.2.

[0318] 4-(7-(5-oxo-2,6-diazaspiro[3.4]octan-6-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (24-B) To a solution of compound 24-A (70 mg, 0.13 mmol) in MeOH (5 mL), HCl-dioxane (4 M, 2 mL) was added and stirred at room temperature overnight. After the reaction was completed, the mixture was concentrated, basified with basic resin, filtered, and concentrated to give the crude product (65 mg, yield: 99%). ESI: [M+H] + = 447.2.

[0319] General Procedure IV 4-(7-(2-methyl-5-oxo-2,6-diazaspiro[3.4]octan-6-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (24) To a solution of compound 24-B (65 mg, 0.15 mmol) in MeOH (5 mL), NaBH3CN (27 mg, 0.45 mmol), HCHO (30 mg) were added and stirred at 40° C. overnight. After the reaction was completed, the mixture was quenched with NH4Cl (aq) and extracted with DCM. The organic layer was separated, dried over anhydrous Na2SO4, filtered and concentrated to dryness. The residue was purified by preparative HPLC to give the desired product (35 mg, yield: 52.4%). ESI: [M+H] + = 461.2; 1 H NMR (400 MHz, MeOH-d4): δ 8.14 (d, J = 2.4 Hz, 1H), 8.08 (d, J = 2.1 Hz, 1H), 8.02 (dd, J = 8.9, 2.0 Hz, 1H), 7.82 - 7.73 (m, 5H), 7.73 - 7.68 (m, 2H), 7.33 (d, J = 1.8 Hz, 1H), 5.42 (s, 2H), 4.59 (dd, J = 28.8, 16.8 Hz, 1H), 4.42 (s, 1H), 4.20 (d, J = 27.9 Hz, 2H), 3.96 (t, J = 7.0 Hz, 2H), 3.08(s, 1.5H), 3.02 (s, 1.5H), 2.61 (s, 2H).

[0320] [ka]

[0321] (S)-tert-Butyl(1-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo [1,2-a][1,4]diazepin-7-yl)-2-oxopyrrolidin-3-yl)carbamate (25-A) Using the procedure described above, 25-A was synthesized. ESI: [M+H] + = 521.2

[0322] (S)-4-(7-(3-amino-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (25-B) To a solution of compound 25-A (60 mg, 0.115 mmol) in dry DCM (2 mL) was added TFA (0.5 mL). The resulting mixture was stirred at room temperature for 2 h and then evaporated to dryness. The residue was redissolved in DCM / MeOH (10:1, 60 mL), washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered and concentrated. The crude product was purified by chromatography on silica gel to give the desired product (30 mg, yield: 62.5%). ESI: [M+H] + = 421.2

[0323] (S)-4-(7-(3-(dimethylamino)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (25) To a solution of compound 25-B (30 mg, 0.07 mmol) in MeOH / DCE (0.5 mL / 0.5 mL) was added HCHO (21 mg, 0.71 mmol) and 1 drop of AcOH. The solution was stirred for 30 min; then NaBH3CN (11 mg, 0.18 mmol) was added. The resulting mixture was stirred at room temperature for an additional 16 h before being diluted with DCM / MeOH (10:1, 60 mL). The solution was washed with water, brine, dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give the desired product (8 mg, yield: 32%). ESI: [M+H] + = 449.2; 1H NMR (400 MHz,MeOH-d4): δ 8.14 (d, J = 2.5 Hz, 1H), 8.03 (d, J = 1.9 Hz, 1H), 7.98 (dd, J = 8.9, 2.5 Hz, 1H), 7.79 -7.65 (m, 5H), 7.29 (d, J = 1.7 Hz, 1H), 5.40 (s, 2H), 4.60 (dd, J = 11.2, 8.7 Hz, 1H), 4.05 (dq, J = 16.3, 9.8 Hz, 2H), 3.03 (s, 6H), 2.79 - 2.61 (m, 1H), 2.57 - 2.39 (m, 1H).

[0324] [ka]

[0325] (S)-4-(7-(4-formyl-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (26-A) To a solution of compound 7-E (870 mg, 2.0 mmol) in DCM (20 mL) was added DMP (1.70 g, 4.0 mmol) in portions at 0° C., and the mixture was then stirred at 20° C. for 3 h. The mixture was then quenched with saturated aqueous NaHCO3 and Na2SO3 solutions; then stirred for 0.5 h and extracted with DCM. The combined organic layers were washed with brine and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography eluting with PE / EA (10 / 1 to 3 / 1) to give the desired compound.

[0326] General Procedure V 4-(7-((4R)-4-((3-fluoropyrrolidin-1-yl)methyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (26) To a solution of compound 26-A (50 mg, 0.12 mmol), TEA (3 drops) and 3-fluoropyrrolidine hydrochloride (60 mg, 0.475 mmol) in dry DCM (5 mL), NaBH(OAc)3 (48.7 mg, 0.23 mmol) was added, and the resulting mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was diluted with DCM (30 mL) and washed with aqueous NaHCO3 and brine. The solution was dried over anhydrous Na2SO4 and filtered; the residue was purified by preparative HPLC to give the desired product (20 mg, yield: 34%). ESI: [M+H] + = 507.2; 1 H NMR (400 MHz, MeOH-d4): δ 8.11 (d, J = 2.4 Hz, 1H), 8.04 (d, J = 2.0 Hz, 1H), 7.85 (dd, J = 8.9, 2.4 Hz, 1H), 7.80- 7.68 (m, 7H), 7.30 (d, J = 1.8 Hz, 1H), 5.39 (s, 2H), 4.20 (ddd, J = 11.2, 8.2, 3.2 Hz, 1H), 3.83 (ddd, J = 9.8, 7.4, 4.9 Hz, 2H), 3.65 - 3.50 (m, 4H), 3.11 - 2.80 (m, 4H), 2.61 (ddd, J = 16.9, 8.6, 2.6 Hz, 2H), 1.42 - 1.31 (m, 1H), 1.17 (t, J = 7.0 Hz, 1H).

[0327] Using the procedures described above, the examples in Table 5 below were synthesized.

[0328] [Table 5]

[0329] [ka]

[0330] cis-tert-Butyl 4-oxohexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate (30-A) To a solution of hexahydropyrrolo[3,4-c]pyrrol-1(2H)-one hydrochloride (50 mg, 0.31 mmol) and TEA (101 mg, 1.0 mmol) in DCM (10 mL) was added Boc2O (65 mg, 0.31 mmol) slowly at 0° C. The resulting mixture was stirred at 20° C. for 2 h. The mixture was diluted with water and extracted with EA. The organic layer was dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated to dryness under reduced pressure. The residue was purified by silica gel chromatography eluting with PE / EA (10 / 1 to 1 / 10) to give the desired product (25 mg, yield: 36.7%). ESI: [M+H] + =227.1

[0331] cis-tert-Butyl 5-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-4-oxohexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate (30-B) Compound 30-B was synthesized using the procedure described above. ESI: [M+H] + = 547.2

[0332] cis-4-(7-(1-oxohexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (30) To a solution of compound 30-B (40 mg, 0.07 mmol) in anhydrous DCM (10 mL) was added TFA (2 mL). The reaction was stirred at 25° C. for 3 h. After consumption of the starting material, the reaction was quenched with saturated NaHCO3 solution and adjusted to pH>7. The resulting mixture was extracted with DCM. The organic phase was washed with brine, dried over Na2SO4, filtered and concentrated to dryness in vacuo. The residue was purified by preparative HPLC to give the desired compound (20 mg, yield: 64.5%). ESI: [M+H] + = 447.2;1 H NMR (400 MHz, MeOH-d4): δ 8.15 (d, J = 2.5 Hz, 1H), 8.07 (d, J = 1.8 Hz, 1H), 7.97 (dd, J = 8.9, 2.5 Hz, 1H), 7.85 - 7.66 (m, 7H), 7.32 (d, J = 1.5 Hz, 1H), 5.41 (s, 2H), 4.30 (dd, J = 10.6, 7.5 Hz, 1H), 3.91 (dd, J = 10.7, 1.5 Hz, 1H), 3.79 - 3.56 (m, 4H), 3.45 - 3.35 (m, 2H).

[0333] [ka]

[0334] tert-Butyl 4-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2- a][1,4]diazepin-7-yl)-3-oxopiperazine-1-carboxylate (31-A) Compound 30-A was synthesized using the procedure described above. ESI: [M+H] + = 521.2

[0335] 4-(7-(2-oxopiperazin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (31-B) To a solution of compound 31-A (78 mg, 0.15 mmol) in MeOH (10 mL) was added HCl in dioxane (2 M, 5 mL) and the mixture was stirred at 25° C. overnight. The reaction mixture was then concentrated in vacuo to give the desired product (50 mg, yield: 79.4%). ESI: [M+H] + = 421.2

[0336] 4-(7-(4-cyclopropyl-2-oxopiperazin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (31) To a solution of compound 31-B (50 mg, 0.12 mmol), TFA (0.5 mL) in DCE (10 mL) was added (1-ethoxycyclopropoxy)trimethylsilane (62.4 mg, 0.35 mmol) and NaBH3CN (8.8 mg, 0.14 mmol). The mixture was stirred at 40° C. for 4 h and then concentrated to dryness under reduced pressure. The residue was purified by preparative HPLC to give the desired product (10 mg, yield: 18.2%). ESI: [M+H] + = 461.2; 1 H NMR (400 MHz, MeOH-d4): δ 8.14 (d, J = 2.1 Hz, 1H), 7.82-7.80 (m, 3H), 7.77-7.69 (m, 5H), 7.63 (dd, J = 8.7, 2.3 Hz, 1H), 7.35 (d, J = 1.8 Hz, 1H), 5.42 (s, 2H), 3.83-3.78 (m, 2H), 3.58 (s, 2H), 3.21-3.16 (m, 2H), 2.06-1.99 (m, 1H), 0.68 - 0.62 (m, 2H), 0.60 (dd, J = 7.4, 3.7 Hz, 2H).

[0337] [ka]

[0338] Methyl (S)-3-((tert-butoxycarbonyl)amino)-4-hydroxybutanoate (32-A) To a solution of (S)-2-((tert-butoxycarbonyl)amino)-4-methoxy-4-oxobutanoic acid (4.95 g, 20 mmol) in 50 mL of EA was added NHS (2.44 g, 21.2 mmol) at 0° C. A solution of DCC (4.17 g, 20.2 mmol) in EA (20 mL) was slowly added to the above solution. Then the mixture was stirred at 30° C. for 15 h. After the reaction was completed, the mixture was washed with saturated aqueous NaHCO3, dried over anhydrous Na2SO4, filtered and concentrated to dryness. To a solution of NaBH4 (1.25 g, 32.6 mmol) in THF (60 mL) and H2O (8 mL) was added dropwise a solution of the above residue in THF (10 mL) at 0° C. The resulting mixture was stirred at 0° C. for 30 min. After the reaction was completed, the mixture was quenched with aqueous NH4Cl and EA. The organic layer was separated, dried over anhydrous Na2SO4, filtered and concentrated to dryness. The residue was purified by chromatography on silica gel eluting with PE / EA (3:1) to give the desired alcohol (3.3 g, yield: 50%). ESI: [M+H] + =234.1

[0339] Methyl (S)-4-azido-3-((tert-butoxycarbonyl)amino)butanoate (32-B) To a solution of compound 31-A (2 g, 8.6 mmol) in DCM (30 mL), TEA (0.85 mL, 6.1 mmol) and MsCl (0.76 mL, 9.86 mmol) were added at 0° C., and the resulting solution was stirred at this temperature for 30 min. After the reaction was completed, the mixture was washed with H2O, dried over anhydrous Na2SO4, filtered, and concentrated in vacuum to obtain crude product (2 g). To a solution of the above product (6.4 mmol) in DMF (30 mL), NaN3 (583 mg, 8.97 mmol) was added, and the resulting mixture was stirred at 80° C. overnight under N2. After the reaction was completed, the mixture was diluted with EA, which was then washed with H2O, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by chromatography on silica gel eluting with PE / EA (8 / 1) to give the desired azide compound (500 mg, yield: 19.5% over two steps). ESI: [M+H] +=259.1

[0340] tert-Butyl (S)-(5-oxopyrrolidin-3-yl)carbamate (32-C) To a solution of compound 32-B (500 mg, 1.9 mmol) in MeOH (10 mL), Pd / C (50 mg, 10%) was added and stirred under H2 at 35°C for 5 h. After the reaction was completed, the mixture was filtered through a celite pad. After the filtrate was concentrated, the residue was purified by chromatography on silica gel (DMC:MeOH=30:1) to give the desired product (250 mg, yield: 65%). ESI: [M+H] + =201.1

[0341] General Procedure VI: tert-Butyl (S)-(1-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo [1,2-a][1,4]diazepin-7-yl)-5-oxopyrrolidin-3-yl)carbamate (32-D) To a solution of compound 32-C (50 mg, 0.25 mmol), compound Int-A (100 mg, 0.25 mmol), Pd(OAc)2 (5.6 mg, 0.025 mmol), BINAP (31 mg, 0.05 mmol) in toluene (4 mL) was added Cs2CO3 (243.7 mg, 0.75 mmol). The resulting mixture was purged with nitrogen and stirred at 110 °C overnight under N2. The mixture was cooled to room temperature, diluted with DCM / MeOH (20 / 1, 200 mL), and filtered. The filtrate was concentrated in vacuo, and the resulting residue was purified by chromatography on silica gel (DCM:MeOH=20:1) to give the desired product as a solid (20 mg, yield: 15%). ESI: [M+H] + =521.2

[0342] (S)-4-(7-(4-amino-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (32) To a solution of compound 32-D (20 mg, 0.04 mmol) in MeOH (5 mL), HCl-dioxane (4 M, 3 mL) was added, and the resulting solution was stirred at room temperature for 3 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was purified by preparative HPLC to give the desired product (16 mg, yield: 99%). ESI: [M+H] + = 421.2; 1 H NMR (400 MHz, CD3OD): δ 8.02 (d, J = 2.2 Hz, 1H), 8.07(d, J = 2.2 Hz, 1H), 7.91 (dd, J = 8.9, 2.5 Hz, 1H), 7.82 - 7.67 (m, 7H), 7.32 (d, J = 1.7 Hz, 1H), 5.41 (s, 2H), 4.43 (dd, J = 11.3, 7.0 Hz, 1H), 4.28 - 4.18 (m, 1H), 3.99 (dd, J = 11.3, 2.6 Hz, 1H), 3.23 - 3.17 (m, 1H), 2.71 (dd, J = 18.1, 3.1 Hz, 1H).

[0343] [ka]

[0344] Benzyl-1-((S)-1-(4-methoxyphenyl)ethyl)-3-methyl-5-oxopyrrolidine-3-carboxylate (33-A) To a solution of compound 7-B' (500 mg, 1.41 mmol) in dry DMF (5 mL) was added 60% NaH (227 mg, 5.67 mmol) and MeI (0.9 mL, 14.16 mmol) at 0 °C under N2. The resulting mixture was stirred at 40 °C for 18 h. Saturated aqueous NH4Cl (20 mL) was slowly added to the above mixture at 0 °C to quench the reaction. The resulting mixture was extracted with EA (30 mL x 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated to dryness. The residue was purified by chromatography on silica gel eluting with PE:EA (10:1 to 3:1) to give the desired product (480 mg, yield: 92.6%). ESI: [M+H] + = 368.2

[0345] 4-(Hydroxymethyl)-1-((S)-1-(4-methoxyphenyl)ethyl)-4-methylpyrrolidin-2-one (33-B) To a solution of compound 33-A (480 mg, 1.65 mmol) in dry MeOH (5 mL) was added NaBH4 (219 mg, 5.77 mmol) at 0 °C. The resulting mixture was stirred at room temperature for 18 h. Saturated aqueous NH4Cl (20 mL) was slowly added to the above mixture at 0 °C to quench the reaction. The resulting mixture was extracted with EA (30 mL x 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated to dryness to give the desired product (380 mg, 87.7% yield) as an off-white solid. ESI: [M+H] + = 264.2

[0346] 4-(Hydroxymethyl)-4-methylpyrrolidin-2-one (33-C) A solution of compound 33-B (380 mg, 1.45 mmol) in TFA (5 mL) was heated to 80 °C and stirred at this temperature for 18 h. The volatile solvents were removed under reduced pressure. The residue was redissolved in MeOH (50 mL) and NH3·H2O was added to the above solution at room temperature. After stirring for 2 h, the reaction mixture was concentrated and the residue was purified by silica gel column chromatography with PE:EA (10:1 to 1:1) to give the desired product (158 mg, yield: 68.7%). ESI: [M+H] + = 130.1

[0347] 4-(7-(4-(hydroxymethyl)-4-methyl-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (33-D) Using the procedure described above, compound 33-D was synthesized as an off-white solid. ESI: [M+H] + = 450.2

[0348] 4-(7-(4-((dimethylamino)methyl)-4-methyl-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (33) Using the procedure described above, compound 33 was synthesized. ESI: [M+H] + = 477.2; 1H NMR (400 MHz, CD3OD): δ 7.97 (d, J = 2.4 Hz, 1H), 7.77 (dd, J = 8.8, 2.5 Hz, 1H), 7.73-7.70 (m, 3H), 7.65 (d, J = 8.5 Hz, 2H), 7.59 (d, J = 8.8 Hz, 1H), 7.54 (d, J = 1.6 Hz, 1H), 7.26 (s, 1H), 7.06 (d, J = 1.6 Hz, 1H), 5.20 (s, 2H), 3.87 (d, J = 9.7 Hz, 1H), 3.64 (d, J = 9.7 Hz, 1H), 2.65 (d, J = 16.8 Hz, 1H), 2.48 (d, J = 2.7 Hz, 2H), 2.40 (s, 1H), 2.35 (s, 1H), 2.34(s, 6H), 1.27 (s, 3H).

[0349] [ka]

[0350] 3-(Hydroxymethyl)pyrrolidin-2-one (34-A) To a solution of ethyl 2-oxopyrrolidine-3-carboxylate (1.5 g, 10.0 mmol) in EtOH (100 mL) was added NaBH4 (2.0 g, 50.0 mmol) and the mixture was stirred at 25° C. for 3 h. Then 4N HCl in MeOH (30 mL) was added to quench the reaction and the resulting solution was stirred for 0.5 h and then concentrated to dryness. The residue was purified by silica gel chromatography to give the desired product (1.0 g, yield: 86.9%). ESI: [M+H] + =116.1

[0351] 4-(7-(3-(hydroxymethyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (34-B) Compound 34-B was synthesized using the procedure described above. ESI: [M+H]+ =436.2; 1 H NMR (400 MHz, CD3OD): δ 8.12 (d, J = 2.5 Hz, 2H), 7.96 (dd, J = 8.9, 2.5 Hz, 1H), 7.83 (s, 1H), 7.80 (s, 1H), 7.76 (d, J = 9.1 Hz, 3H), 7.71 (d, J = 8.6 Hz, 2H), 7.35 (s, 1H), 5.42 (s, 2H), 3.95 (dt, J = 6.1, 3.9 Hz, 3H), 3.82 (dd, J = 10.9, 3.7 Hz, 1H), 2.86 (dq, J = 13.0, 4.2 Hz, 1H), 2.37 (ddd, J = 15.0, 12.2, 5.8 Hz, 1H), 2.22 (dq, J = 12.7, 8.3 Hz, 1H).

[0352] 4-(7-(3-((dimethylamino)methyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (34) Using the procedure described above, compound 34 was synthesized. ESI: [M+H] + = 463.2; 1 H NMR (400 MHz, CD3OD): δ 8.08 (d, J = 2.5 Hz, 2H), 8.03 (dd, J = 8.9, 2.5 Hz, 1H), 7.81 - 7.69 (m, 7H), 7.34 (s, 1H), 5.41 (s, 2H), 4.06 - 3.99 (m, 2H), 3.69 - 3.59 (m, 1H), 3.38 (dd, J = 13.5, 5.0 Hz, 2H), 2.58 - 2.48 (m, 1H), 2.01 (tt, J = 12.6, 9.0 Hz, 1H). Compound 34 was separated by SFC on a chiral preparative column to give two enantiomers 35 and 36, whose stereochemistry was not determined. The first eluate was arbitrarily assigned as the S-isomer (740 mg) and the second eluate was assigned as the R-isomer (700 mg). ESI: [M+H]+ = 463.2; Peak 1 1 H NMR (400 MHz, CD3OD): δ 7.97 (d, J = 2.5 Hz, 1H), 7.79 (dd, J = 8.8, 2.5 Hz, 1H), 7.73 - 7.67 (m, 3H), 7.67 - 7.62 (m, 2H), 7.58 (d, J = 8.8 Hz, 1H), 7.53 (d, J = 1.9 Hz, 1H), 7.25 (d, J = 1.5 Hz, 1H), 7.05 (d, J = 1.9 Hz, 1H), 5.18 (s, 2H), 3.96 - 3.85 (m, 2H), 2.93 (ddd, J = 18.3, 9.6, 3.9 Hz, 1H), 2.84 (dd, J = 12.5, 3.9 Hz, 1H), 2.58 (dd, J = 12.4, 9.7 Hz, 1H), 2.52 - 2.41 (m, 1H), 2.33 (s, 6H), 2.01 (dq, J = 12.8, 9.0 Hz, 1H). A solution of the hydrochloride salt of compound 35 (400 mg, 0.86 mmol) in dioxane (5 mL, 5 mmol) was stirred at room temperature for 1 h. The reaction mixture was then concentrated in vacuo to give the hydrochloride salt as a white solid (440 mg, yield: 99%). 1 H NMR (400 MHz, CD3OD): δ 8.17 (d, J = 2.2 Hz, 1H), 8.12 (d, J = 2.5 Hz, 1H), 8.07 (dd, J = 8.9, 2.5 Hz, 1H), 7.87 (dd, J = 3.9, 2.0 Hz, 2H), 7.77 (dt, J = 19.5, 8.8 Hz, 5H), 7.39 (d, J = 1.8 Hz, 1H), 5.46 (d, J = 9.7 Hz, 2H), 4.10 - 3.98 (m, 2H), 3.76 - 3.69 (m, 1H), 3.69 - 3.62 (m, 2H), 3.58 (m, 1H), 3.00 (d, J = 9.9 Hz, 6H), 2.55 (d, J = 3.9 Hz, 1H), 2.10 - 1.94 (m, 1H). ピーク2 1 H NMR (400 MHz, CD3OD): δ 7.97 (d, J = 2.5 Hz, 1H), 7.80 (dd, J = 8.8, 2.5 Hz, 1H), 7.73 - 7.67 (m, 3H), 7.67 - 7.62 (m, 2H), 7.53 (d, J = 1.9 Hz, 1H), 7.25 (d, J = 1.5 Hz, 1H), 7.05 (d, J = 1.9 Hz, 1H), 5.18 (s, 2H), 3.97 - 3.86 (m, 2H), 2.93 (ddd, J = 18.3, 9.6, 3.9 Hz, 1H), 2.84 (dd, J = 12.5, 4.0 Hz, 1H), 2.58 (dd, J = 12.4, 9.7 Hz, 1H), 2.52 - 2.41 (m, 1H), 2.33 (s, 6H), 2.01 (dq, J = 12.6, 8.9 Hz, 1H).

[0353]

change

[0354] The above-mentioned formula is made of して and compound 37 is synthesized by した. ESI: [M+H] + =493.2; 1 H-NMR (400 MHz, CD3OD): δ 8.10 (d, J = 2.4 Hz, 1H), 8.03 (s, 1H), 7.98 (dd, J = 8.9, 2.4 Hz, 1H), 7.77- 7.70 (m, 7H), 7.30 (s, 1H), 5.39 (s, 2H), 4.02 (d, J = 7.9 Hz, 2H), 3.93 (t, J = 5.0 Hz, 2H), 3.80-3.70 (m, 1H), 3.60-3.50 (m, 1H), 3.43-3.41 (m, 2H), 3.06 (s, 3H), 2.55-2.53 (m, 1H), 2.13 - 1.90 (m, 1H).

[0355]

change

[0356] 3-(Hydroxydeuterated methyl)pyrrolidin-2-one (38-A) To a solution of ethyl 2-oxopyrrolidine-3-carboxylate (300 mg, 1.91 mmol) in CD3OD (3 mL) was added NaBD4 (280 mg, 6.69 mmol) at 0°C. The resulting mixture was stirred at room temperature for 16 h under argon atmosphere. To the reaction solution was added saturated NH4Cl (5 mL) at 0°C, then extracted with DCM / MeOH (10:1, 30 mL*3). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered and concentrated. The residue was purified by chromatography on silica gel to give the desired compound as an off-white solid (160 mg, yield: 71.4%). ESI: [M+H] + = 118.1; 1 H-NMR (400 MHz, CDCl3): δ 5.94 (s, 1H), 3.48 - 3.31 (m, 2H), 2.90 (s, 1H), 2.34 - 2.17 (m, 1H), 2.07 - 1.85 (m, 1H).

[0357] 4-(7-(3-(hydroxydeuterated methyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (38-B) Compound 38-B was synthesized using the procedure described above. ESI: [M+H] + = 438.2

[0358] 4-(7-(3-(aminodeuterated methyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (38-C) Compound 33-C was synthesized using the procedure described above. ESI: [M+H] + = 437.2

[0359] 4-(7-(3-((di-deuteromethylamino)deuteromethyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (038) To a solution of compound 38-C (18 mg, 0.12 mmol) in dry CD3OD / DCE (0.3 mL / 0.3 mL) was added CD2O (13 mg, 0.41 mmol) and cat.AcOH (2 mg). The resulting solution was stirred for 30 min before the addition of NaBD3CN (8 mg, 0.12 mmol). The resulting mixture was stirred at room temperature for 4 h and then diluted with DCM / MeOH (10:1, 60 mL). The solution was washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative TLC to give the desired product (15 mg, yield: 78.9%). ESI: [M+H] + = 471.3; 1 H NMR (400 MHz, MeOH-d4): δ 8.06(d, J = 2.4 Hz, 1H), 8.02-7.99 (m, 2H), 7.75-7.65 (m, 7H), 7.30 (d, J = 1.6 Hz, 1H), 5.39 (s, 2H), 4.18-3.86 (m, 2H), 3.42-3.35 (m, 1H), 2.53 (ddd, J = 8.8, 7.5, 2.9 Hz, 1H), 2.17-1.87 (m, 1H).

[0360] [ka]

[0361] Ethyl 3-methyl-2-oxopyrrolidine-3-carboxylate (39-A) To a solution of ethyl 2-oxopyrrolidine-3-carboxylate (500 mg, 3.2 mmol) in dry THF (5 mL) was added n-BuLi (2.5 M in THF, 1.4 mL, 3.5 mmol) dropwise at 0 °C and the resulting mixture was allowed to warm slowly to room temperature before cooling again to -78 °C. CHI (497 mg, 3.5 mmol) was added dropwise to the above solution at -78 °C. The reaction solution was allowed to warm to room temperature and continued to stir for an additional 16 h under argon atmosphere. To the above solution was added saturated aqueous NH4Cl dropwise at 0 °C. The resulting mixture was extracted with DCM / MeOH (10:1, 30 mL*3). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered and concentrated to dryness. The residue was purified by chromatography on silica gel to give the desired compound (350 mg, yield: 64.2%) as an oil. ESI: [M+H] + = 172.1; 1 H-NMR (400 MHz, CDCl3): δ 6.00 (s, 1H), 4.26 - 4.16 (m, 2H), 3.48 (dt, J = 9.1, 7.4 Hz, 1H), 3.35 (td, J = 8.8, 4.0 Hz, 1H), 2.64 (ddd, J = 12.9, 7.8, 4.0 Hz, 1H), 2.04 (ddd, J = 13.0, 8.4, 7.0 Hz, 1H), 1.45 (d, J = 5.0 Hz, 3H), 1.28 (dd, J = 9.5, 4.8 Hz, 3H).

[0362] 3-(Hydroxymethyl)-3-methylpyrrolidin-2-one (39-B) To a solution of compound 39-A (350 mg, 2.05 mmol) in EtOH (10 mL) was added NaBH4 (272 mg, 7.16 mmol) at 0°C. The resulting mixture was stirred at room temperature under argon gas for 48 h. To the above solution was added saturated aqueous NH4Cl dropwise at 0°C. The resulting mixture was extracted with DCM (30 mL*3). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered and concentrated to dryness. The residue was purified by chromatography on silica gel to give the desired compound (234 mg, yield: 88.6%) as an oil. ESI: [M+H] + = 130.1; 1 H NMR (400 MHz, CDCl3): δ 5.97 (s, 1H), 3.67 (d, J = 10.9 Hz, 1H), 3.55 (d, J = 10.9 Hz, 1H), 3.44 - 3.30 (m, 2H), 2.50 (s, 1H), 2.23 (dt, J = 12.7, 8.2 Hz, 1H), 1.86 (ddd, J = 12.7, 7.1, 4.3 Hz, 1H), 1.20 (s, 3H).

[0363] 4-(7-(3-(hydroxymethyl)-3-methyl-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (39-C) Compound 39-C was synthesized using the procedure described above. ESI: [M+H] + = 450.2

[0364] 4-(7-(3-((dimethylamino)methyl)-3-methyl-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (39) Using the procedure described above, compound 39 was synthesized. ESI: [M+H] + = 477.2; 1H NMR (400 MHz, CD3OD): δ 8.12 (d, J = 2.4 Hz, 1H), 8.06 (s, 1H), 7.99 (dd, J = 8.9, 2.5 Hz, 1H), 7.79 - 7.67 (m, 7H), 7.31 (d, J = 1.3 Hz, 1H), 5.40 (s, 2H), 4.05 (ddd, J = 18.7, 13.0, 5.1 Hz, 2H), 3.52 (q, J = 13.8 Hz, 2H), 3.00 (s, 6H), 2.37 - 2.14 (m, 2H), 1.42 (s, 3H).

[0365] [ka]

[0366] 3-(2-Nitroethyl)dihydrofuran-2(3H)-one (40-A) A 250 mL round bottom flask was charged with 3-methylideneoxolan-2-one (5 g, 50.97 mmol), DBU (1 g, 6.57 mmol), and CH3NO2 (100 mL). The resulting solution was stirred at 25° C. until completion. The resulting mixture was concentrated under vacuum, then taken up in 100 mL of DCM and washed successively with 3.0 M HCl, water, saturated aqueous NaHCO3, and brine. The organic solution was dried over anhydrous sodium sulfate and then concentrated under vacuum to give the desired compound (5.8 g, yield: 99%). ESI: [M+H] + = 160.1

[0367] 3-(2-Hydroxyethyl)pyrrolidin-2-one (40-B) A 250 mL round bottom flask was charged with compound 40-A (5.8 g, 36.47 mmol), Raney Ni (4.44 g), methanol (100 mL), and magnesium sulfate (4 g). The resulting solution was stirred at room temperature under hydrogen atmosphere until completion. The solids were filtered off and the filtrate was concentrated under vacuum to give the desired compound (2.3 g, yield: 48.5%). ESI: [M+H] + = 130.1

[0368] 4-(7-(3-(2-hydroxyethyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (40-C) Compound 40-C was synthesized using the procedure described above. ESI: [M+H] + = 450.2

[0369] 4-(7-(3-(2-(dimethylamino)ethyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (40) Using the procedure described above, compound 40 was synthesized. ESI: [M+H] + = 477.2; 1 H NMR (400 MHz, CD3OD): δ 8.09-8.08 (m, 2H), 7.98 (dd, J = 8.9, 2.5 Hz, 1H), 7.80 (d, J = 1.7 Hz, 1H), 7.79 - 7.73 (m, 4H), 7.70 (d, J = 8.5 Hz, 2H), 7.34 (d, J = 1.7 Hz, 1H), 5.41 (s, 2H), 3.98 (dd, J = 9.3, 4.7 Hz, 2H), 3.44 (ddd, J = 12.8, 9.7, 6.5 Hz, 1H), 3.27 (dd, J = 8.9, 4.1 Hz, 1H), 2.95 (s, 6H), 2.91 - 2.79 (m, 1H), 2.47 (qd, J = 8.8, 4.5 Hz, 1H), 2.26 (ddt, J = 13.7, 9.8, 6.8 Hz, 1H), 2.08 - 1.84 (m, 2H). Compound 40 was separated by SFC on a chiral preparative column to give two enantiomers 41 and 42, whose stereochemistry was not determined. The first eluent was assigned as the S-isomer and the second eluent was assigned as the R-isomer.

[0370] (S)-4-(7-(3-(2-(dimethylamino)ethyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (41) 1 H NMR (400 MHz, CD3OD): δ 7.98 (d, J = 2.5 Hz, 1H), 7.82 (dd, J = 8.8, 2.5 Hz, 1H), 7.73 - 7.62 (m, 5H), 7.59 (d, J = 8.8 Hz, 1H), 7.53 (d, J = 1.9 Hz, 1H), 7.26 (d, J = 1.5 Hz, 1H), 7.06 (d, J = 1.9 Hz, 1H), 5.19 (s, 2H), 3.99 - 3.84 (m, 2H), 3.04-2.92 (m, 1H), 2.91 - 2.72 (m, 2H), 2.62 (s, 6H), 2.50 - 2.36 (m, 1H), 2.18 (ddt, J = 12.8, 9.9, 6.3 Hz, 1H), 2.00 - 1.70 (m, 2H).

[0371] (R)-4-(7-(3-(2-(dimethylamino)ethyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (42) 1H NMR (400 MHz, CD3OD): δ 7.98 (d, J = 2.4 Hz, 1H), 7.79 (dd, J = 8.8, 2.5 Hz, 1H), 7.69-7.66 (m, 5H), 7.58 (d, J = 8.8 Hz, 1H), 7.54 (d, J = 1.8 Hz, 1H), 7.25 (d, J = 1.4 Hz, 1H), 7.05 (d, J = 1.9 Hz, 1H), 5.19 (s, 2H), 3.97 - 3.83 (m, 2H), 2.71 (ddd, J = 18.4, 9.1, 4.8 Hz, 1H), 2.62 - 2.37 (m, 4H), 2.31 (s, 6H), 2.14 (ddd, J = 21.3, 10.7, 5.9 Hz, 1H), 1.87 (dq, J = 12.4, 8.9 Hz, 1H), 1.73 - 1.53 (m, 1H).

[0372] [ka]

[0373] 4-(7-(3-methylene-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (43-A) To a 0°C solution of compound 34-B (1.31 g, 3.0 mmol), TEA (909 mg, 9.0 mmol) in DCM (15 mL) was added MsCl (689 mg, 6.00 mmol) and stirred for 1 h. The reaction was quenched with aqueous NaHCO3; the resulting mixture was extracted with DCM three times. The combined organic layers were dried over Na2SO4, filtered, and concentrated to dryness. The residue was then purified by chromatography on silica gel to give the desired product (1.22 g, yield: 79.3%), which was used directly. At room temperature, a solution of the aforementioned compound (0.85 g, 1.66 mmol), TEA (3 mL) in DMF (10 mL) was stirred for 5 h. The reaction solution was dried under reduced pressure, and the residue was purified by chromatography on silica gel to give the desired compound (0.52 g, yield: 63.4%). ESI: [M+H] + =418.2

[0374] (R)-4-(7-(3-(2-methyl-2-nitropropyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (43-B) A solution of compound 43-A (0.52 g, 1.32 mmol), 2-nitropropane (0.59 g, 6.6 mmol), DBU (1.00 g, 6.6 mmol) in THF (15 mL) was stirred under reflux overnight under N2. The reaction mixture was concentrated under reduced pressure and the residue was purified by chromatography on silica gel to give the desired product (0.4 g, yield: 59.8%). ESI: [M+H] + =507.2

[0375] (R)-4-(7-(3-(2-amino-2-methylpropyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (43) To a solution of compound 43-B (50 mg, 0.10 mmol), NH4Cl (26 mg, 0.50 mmol) in EtOH (8 mL), THF (4 mL), HO (4 mL) was added Fe (28 mg, 0.50 mmol). The reaction mixture was stirred at 80 °C overnight under N2. The reaction mixture was cooled to room temperature and filtered through a celite pad. The filtrate was concentrated and the residue was purified by flash chromatography followed by preparative HPLC to give the desired product (14 mg, yield: 29.2%). ESI: [M+H] + = 477.2; 1 H NMR (400 MHz, CD3OD): δ 8.10 (s, 1H), 8.09 (s, 1H), 7.99 (dd, J = 8.9, 2.5 Hz, 1H), 7.81 - 7.69 (m, 7H), 7.34 (d, J = 1.8 Hz, 1H), 5.42 (s, 2H), 4.02 - 3.93 (m, 2H), 3.07 - 2.98 (m, 1H), 2.53 (s, 1H), 2.30 - 2.21 (m, 1H), 1.96 (dd, J = 19.0, 7.0 Hz, 1H), 1.80 (dd, J = 14.9, 6.1 Hz, 1H), 1.45 (s, 3H), 1.44 (s, 3H).

[0376] [ka]

[0377] tert-Butyl 3-(2-(1,1-dimethylethylsulfinamido)propan-2-yl)-2-oxopyrrolidine-1-carboxylate (44-A) To a solution of tert-butyl 2-oxopyrrolidine-1-carboxylate (926.1 mg, 5 mmol) in THF (20 mL) was added NaHMDS (2M, 3 mL) at -78 °C. The mixture was stirred at -78 °C for 0.5 h, then a solution of 2-methyl-N-propan-2-ylidene-propane-2-sulfinamide (806.3 mg, 5 mmol) in THF (2 mL) was added at -78 °C. The resulting solution was stirred at -78 °C for another 0.5 h, and the reaction was quenched by addition of saturated aqueous NH4Cl solution. The mixture was extracted three times with EA. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure to give a crude oil, which was purified by flash chromatography eluting with PE / EA (10 / 1 to 1 / 1) to give the desired product (470 mg, yield: 27.1%) as a white solid. ESI: [M+H] + = 347.2

[0378] tert-Butyl (2-(2-oxopyrrolidin-3-yl)propan-2-yl)carbamate (44-B) A solution of compound 44-A (447 mg, 1.29 mmol) in HCl (4N in dioxane, 3 mL) was stirred at 20° C. for 2 h and then concentrated under reduced pressure to give the crude compound as HCl salt, which was used directly in the next step. To a solution of the above crude compound (185 mg, 1.3 mmol) and TEA (394 mg, 3.9 mmol) was added Boc2O (283 mg, 1.3 mmol). The solution was stirred at 20° C. for 2 h before the addition of H2O. The resulting mixture was extracted three times with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and concentrated to dryness under reduced pressure. The residue was purified by flash chromatography eluting with DCM / MeOH (100 / 1 to 20 / 1) to give the desired compound (190 mg, yield: 60.3%). ESI: [M+H] + = 243.2

[0379] tert-Butyl (2-(1-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-2-oxopyrrolidin-3-yl)propan-2-yl)carbamate (44-C) Compound 44-C was synthesized using the procedure described above. ESI: [M+H] + = 563.3

[0380] 4-(7-(3-(2-aminopropan-2-yl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (44) To a solution of compound 44-C (40.00 mg, 0.07 mmol) in DCM (10 mL), TFA (2 mL) was added and the resulting solution was stirred at 20° C. for 18 h. To the above solution, saturated aqueous NaHCO3 (20 mL) was slowly added; the resulting mixture was extracted with DCM three times. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give the desired product (12 mg, yield: 36.5%). ESI: [M+H] + = 463.2; 1 H-NMR (400 MHz, CD3OD): δ 8.07 (s, 1H), 8.00 (m, 2H), 7.71 (m, 7H), 7.29 (d, J = 4.2 Hz, 1H), 5.38 (s, 2H), 4.11 - 3.82 (m, 2H), 3.08 (m, 1H), 2.47 - 2.34 (m, 1H), 2.18 - 2.01 (m, 1H), 1.44 (s, 3H), 1.44 (s, 3H).

[0381] [ka]

[0382] 3-(tert-butyl) 6-ethyl (1S,5R,6R)-2-oxo-3-azabicyclo[3.1.0]hexane-3,6-dicarboxylate (45-A) A mixture of NaIO4 (1.68 g, 7.84 mmol) and RuCl3.3H2O (51 mg, 0.196 mmol) in water (20 mL) was stirred at room temperature for 30 min. 3-(tert-butyl) 6-ethyl (1R,5S,6s)-3-azabicyclo[3.1.0]hexane-3,6-dicarboxylate (1 g, 3.92 mmol) in EA (20 mL) was added to the above mixture. The resulting mixture was stirred at room temperature for 3 h and filtered. The EA layer in the filtrate was separated and concentrated to dryness. The residue was purified by silica gel chromatography eluting with PE / EA (10 / 1 to 3 / 1) to give the desired product (520 mg, yield: 49%). ESI: [M+H] + =270.1

[0383] Ethyl (1S,5R,6R)-2-oxo-3-azabicyclo[3.1.0]hexane-6-carboxylate (45-B) To a solution of compound 45-A (800 mg, 2.97 mmol) in DCM (20 mL) was added HCl in dioxane (4 M, 2 mL) at room temperature and stirred for 1 h. The reaction solution was then concentrated to dryness to give the desired compound (500 mg, yield: 99%) as a solid.

[0384] (1S,5S,6R)-6-(hydroxymethyl)-3-azabicyclo[3.1.0]hexan-2-one (45-C) To a solution of compound 45-B in dry THF was added DIBAL-H (1.5M in toluene, 4 mL) at 0° C. under N2. The resulting reaction solution was stirred at 0° C. to room temperature overnight before being slowly quenched with water. After filtration of the resulting mixture, the filtrate was concentrated to give the crude compound, which was purified by chromatography on silica gel (DCM:MeOH=10:1) to give the desired compound (120 mg, yield: 32%) as a yellow solid. ESI: [M+H] + =128.1

[0385] 4-(7-((1S,5S,6R)-6-(hydroxymethyl)-2-oxo-3-azabicyclo[3.1.0]hexan-3-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (45-D) Compound 45-D was synthesized using the procedure described above. ESI: [M+H] + = 448.2

[0386] 4-(7-((1R,5S,6R)-6-((dimethylamino)methyl)-2-oxo-3-azabicyclo[3.1.0]hexan-3-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (45) Using the procedure described above, compound 45 was synthesized. ESI: [M+H]+ =475.2; 1 H NMR (400 MHz, CD3OD): δ 8.25 - 8.02 (m, 2H), 7.93 (dd, J = 8.9, 2.3 Hz, 1H), 7.83 - 7.63 (m, 7H), 7.33 (s, 1H), 5.40 (s, 2H), 4.28 (dd, J = 11.6, 6.5 Hz, 1H), 3.92 (d, J = 11.5 Hz, 1H), 3.42 (dd, J = 13.6, 5.4 Hz, 1H), 3.03 - 2.86 (m, 7H), 2.63 - 2.41 (m, 2H), 1.82 (dd, J = 8.2, 5.5 Hz, 1H).

[0387] Using the procedure described above, compound 46 in Table 6 was synthesized.

[0388] [Table 6]

[0389] [ka]

[0390] tert-Butyl 3-methylene-2-oxopyrrolidine-1-carboxylate (47-A) To a solution of LiHMDS (1.0M in THF, 55.4 mL, 55.4 mmol) in dry THF (10 mL) was added dropwise a solution of tert-butyl 2-oxopyrrolidine-1-carboxylate (5.0 g, 27.03 mmol) in dry THF (10 mL) at -78 °C. The resulting mixture was stirred at room temperature for 40 min. Then 2,2,2-trifluoroethyl 2,2,2-trifluoroacetate (6.9 mL, 51.36 mmol) was added dropwise at room temperature and the resulting mixture was stirred at room temperature for another 20 min. After consumption of the starting material, the reaction was quenched with saturated aqueous NH4Cl solution at 0 °C and extracted with EA (30 mL * 3). The combined organic layers were washed with water, brine, dried over Na2SO4, filtered and concentrated. The crude product was dissolved in toluene (100 mL) and (CHO) n (4.1 g, 135.2 mmol) and K2CO3 (8.2 g, 59.5 mmol) were added to the above solution. The resulting mixture was stirred at 108 °C for 2 h. After consumption of the starting material, the reaction mixture was cooled to room temperature, diluted with water and extracted with EA (30 mL * 3). The combined organic layers were washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel to give the desired product (1.1 g, yield: 20.4%) as a solid. ESI: [M+H] + = 198; 1 H NMR (400 MHz, CDCl3): δ 6.19 (t, J = 2.8 Hz, 1H), 5.48 (t, J = 2.4 Hz, 1H), 3.82 - 3.65 (m, 2H), 2.75 (tt, J = 7.1, 2.7 Hz, 2H), 1.55 (s, 9H).

[0391] 5-(tert-butyl) 1-methyl 4-oxo-5-azaspiro[2.4]heptane-1,5-dicarboxylate (47-B) To a solution of methyl 2-bromoacetate (1.3 mL, 14.47 mmol) in dry MeCN (20 mL) was added DABCO (1.62 g, 14.47 mmol), and after 30 min, a solution of Cs2CO3 (4.71 g, 14.47 mmol) and compound 47-A (950 mg, 4.82 mmol) in dry MeCN (35 mL) was added. The resulting mixture was stirred at 80 °C for 22 h. The reaction mixture was cooled to room temperature, diluted with water, and extracted with EA (30 mL * 3). The combined organic layers were washed with water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel to give the desired product (300 mg, yield: 23%) as a solid. ESI [M-56 + H] + = 214; 1 H NMR (400 MHz, CDCl3): δ 3.86 - 3.78 (m, 2H), 3.76 - 3.65 (m, 3H), 2.40 - 2.26 (m, 1H), 2.25 - 2.16 (m, 2H), 1.69 (s, 2H), 1.61 (dd, J = 8.9, 4.1 Hz, 1H), 1.57 - 1.49 (m, 9H), 1.43 (dd, J = 6.1, 3.9 Hz, 2H), 1.31 - 1.22 (m, 1H).

[0392] Methyl 4-oxo-5-azaspiro[2.4]heptane-1-carboxylate (47-C) To a solution of compound 47-B (300 mg, 1.11 mmol) in dry DCM (3 mL) was added TFA (0.5 mL) dropwise at 0° C. The resulting mixture was stirred at room temperature for 2 h and then concentrated to dryness. The residue was purified by column chromatography on silica gel to give the desired product (174 mg, yield: 92.5%) as a solid. ESI [M-56+H] + = 170.

[0393] 1-(Hydroxymethyl)-5-azaspiro[2.4]heptan-4-one (47-D) To a solution of compound 47-C (170 mg, 0.92 mmol) in dry THF (3 mL) was added LiBHEt3 (1.0 M in THF, 4.6 mL, 4.62 mmol) dropwise at -78 °C. The resulting mixture was stirred at -78 °C for 2 h. After that time, the reaction was quenched with saturated aqueous NH4Cl at 0 °C and extracted with EA (30 mL * 3). The combined organic layers were washed with water, brine, dried over Na2SO4, filtered and concentrated to dryness. The residue was purified by column chromatography on silica gel to give the desired product (97 mg, yield: 74.6%) as an oil. ESI [M-56 + H] + = 142; 1 H NMR (400 MHz, MeOH-d4) δ 3.79 (dd, J = 11.7, 5.4 Hz, 1H), 3.48 (dd, J = 7.9, 6.8 Hz, 2H), 3.34 (s, 2H), 2.45 - 2.29 (m, 1H), 2.10 (ddd, J = 13.0, 12.3, 8.3 Hz, 1H), 1.62 - 1.48 (m, 1H), 1.14 (dd, J = 9.2, 4.3 Hz, 1H), 0.62 (dd, J = 6.3, 4.4 Hz, 1H).

[0394] 4-(7-(1-(hydroxymethyl)-4-oxo-5-azaspiro[2.4]heptan-5-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (47-E) Compound 47-E was synthesized using the procedure described above. ESI: [M+H] + = 462

[0395] 4-(7-(1-((dimethylamino)methyl)-4-oxo-5-azaspiro[2.4]heptan-5-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (47) Using the procedure described above, compound 47 was synthesized. ESI: [M+H] + = 489; 1H NMR (400 MHz, MeOH-d4): δ 8.09 (d, J = 2.4 Hz, 1H), 7.99 (d, J = 1.9 Hz, 1H), 7.94 (dd, J = 8.9, 2.5 Hz, 1H), 7.73-7.70 (m, 6H), 7.65 (d, J = 1.8 Hz, 1H), 7.27 (d, J = 1.8 Hz, 1H), 5.36 (s, 2H), 4.10 (dt, J = 9.6, 6.5 Hz, 2H), 3.54 (dd, J = 13.2, 4.9 Hz, 1H), 3.04 - 2.93 (m, 7H), 2.39 (dt, J = 13.0, 9.0 Hz, 1H), 2.26 (ddd, J = 12.8, 8.3, 4.6 Hz, 1H), 1.82 - 1.69 (m, 1H), 1.50 (dd, J = 8.7, 4.7 Hz, 1H), 1.11 - 1.01 (m, 1H).

[0396] [ka]

[0397] Ethyl (1S,2S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-1-(((methylsulfonyl)oxy)methyl)cyclopropane-1-carboxylate (48-B) To a solution of compound 48-A (1.4 g, 4.86 mmol) in DCM (5 mL) was added TEA (1 mL) and MsCl (1.1 g, 9.72 mmol) at 0° C. The resulting mixture was stirred for 1 h and then poured into NaHCO3(aq) (20 mL). The mixture was extracted with DCM (2×20 mL). The combined organic layers were washed with brine (2×20 mL), dried over Na2SO4, filtered, and concentrated to dryness. The crude product was purified by chromatography on silica gel eluting with DCM:MeOH (20:1) to give the desired product (1.7 g, yield: 98%).

[0398] Ethyl (1R,2S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-1-(cyanomethyl)cyclopropane-1-carboxylate (48-C) To a solution of compound 48-B (1.7 g, 4.6 mmol) in DMSO (5 mL) was added NaCN (455 mg, 9.29 mmol). The mixture was stirred at 95° C. for 3 h. The reaction mixture was cooled to room temperature, poured into NaHCO3 (20 mL), and extracted with DCM (20 mL x 2). The combined organic layers were washed with brine (20 mL x 2), dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was purified by chromatography on silica gel to give the desired product (800 mg, yield: 61.5%).

[0399] (1S,3R)-1-(((tert-butyldimethylsilyl)oxy)methyl)-5-azaspiro[2.4]heptan-4-one (48-D) To a solution of compound 48-C (400 mg, 1.34 mmol) in MeOH (5 mL) was added Raney Ni (455 mg, 9.29 mmol) and the resulting mixture was stirred under H2 at 65 °C for 3 h. The reaction mixture was cooled to room temperature, filtered through a celite pad and concentrated to dryness. The residue was purified by eluting silica gel chromatography to give the desired product (100 mg, yield: 29.15%). ESI: [M+H] + = 256.2

[0400] 4-(7-((1S,3R)-1-(((tert-butyldimethylsilyl)oxy)methyl)-4-oxo-5-azaspiro[2.4]heptan-5-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (48-E) Compound 48-E was synthesized using the procedure described above. ESI: [M+H] + =576.3

[0401] 4-(7-((1S,3R)-1-(hydroxymethyl)-4-oxo-5-azaspiro[2.4]heptan-5-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (48-F) To a solution of compound 48-E (80 mg, 0.14 mmol) in THF (2 mL) was added TBAF (1 M in THF, 0.5 mL) at room temperature. The mixture was stirred for 3 h and then poured into H2O (20 mL). The resulting mixture was extracted with DCM (2 x 20 mL). The combined organic layers were washed with brine (2 x 20 mL), dried over Na2SO4, filtered and concentrated to dryness. The crude product was purified by chromatography on silica gel eluting to give the desired product (60 mg, yield: 93.75%). ESI: [M+H] + =462.2

[0402] 4-(7-((1S,3R)-1-((dimethylamino)methyl)-4-oxo-5-azaspiro[2.4]heptan-5-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (48) Using the procedure described above, compound 48 was synthesized. ESI: [M+H] + = 489.2; 1 H NMR (400 MHz, MeOH-d4) δ 8.10 (d, J = 2.4 Hz, 1H), 8.03 (s, 1H), 7.95 (dd, J = 8.9, 2.5 Hz, 1H), 7.84 - 7.64 (m, 7H), 7.30 (s, 1H), 5.38 (s, 2H), 4.16-4.09 (m, 2H), 3.54 (dd, J = 13.2, 4.9 Hz, 1H), 2.47 - 2.33 (m, 1H), 2.29-2.23 (m, 1H), 1.84 - 1.66 (m, 1H), 1.50 (dd, J = 8.7, 4.6 Hz, 1H), 1.12 - 1.03 (m, 1H).

[0403] [ka]

[0404] Ethyl (R)-ethyl 1-(2-((1-(4-methoxyphenyl)ethyl)amino)acetyl)cyclopropanecarboxylate (49-A) To a solution of (R)-1-(4-methoxyphenyl)ethan-1-amine (6.45 g, 42.74 mmol) in CH3CN (50 mL) was added a solution of ethyl 1-(2-bromoacetyl)cyclopropane-1-carboxylate (10 g, 42.74 mmol), TEA (6.5 mL, 47 mmol) in CH3CN (10 mL) dropwise at 0 °C. The mixture was stirred under N2 at 0 °C for 3 h. After the reaction was complete, the mixture was concentrated and redissolved in EA (200 mL). The organic solution was extracted with 1N HCl (200 mL) and the aqueous solution was washed twice with EA before being neutralized with 1N NaOH (200 mL). The aqueous solution was extracted with EA. The organic layer was separated, dried over anhydrous Na2SO4, filtered and concentrated to give the desired product (6 g, yield: 45.9%). ESI: [M+H] + =306.2

[0405] (R)-Ethyl 1-(2-(2-(diethoxyphosphoryl)-N-(1-(4-methoxyphenyl)ethyl)acetamido)acetyl)cyclopropanecarboxylate (49-B) To a solution of 2-(diethoxyphosphoryl)acetic acid (4.6 g, 23.6 mmol) in THF (50 mL), CDI (4.1 g, 25.57 mmol) was added and stirred at 0 °C for 1 h. Compound 49-A (6 g, 19.67 mmol) in THF (10 mL) was added dropwise to the above solution at 0 °C, and the mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was diluted with EA and washed with H2O, 1N HCl (100 mL), NaHCO3 (100 mL) and brine. The organic layer was dried over anhydrous Na2SO4, filtered and concentrated to dryness. The residue was purified by chromatography on silica gel eluted with PE:EA (10:1 to 1:1) to give the desired product (3.2 g, yield: 25.8%). ESI: [M+H] + =484.2.

[0406] (R)-Ethyl 1-(1-(1-(4-methoxyphenyl)ethyl)-5-oxo-2,5-dihydro-1H-pyrrol-3-yl)cyclopropanecarboxylate (49-C) To a solution of compound 49-B (3.2 g, 6.6 mmol) in toluene (30 mL) was added t-BuOK (1.48 g, 13.2 mmol) at 0° C., and the mixture was stirred at room temperature for 1 h. Citric acid (aq) was added dropwise at 0° C. to adjust pH<7, then extracted with EA three times. The combined organic layers were washed with NaHCO3, brine, dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by chromatography on silica gel eluting with PE:EA (10:1 to 2:1) to give the desired product (1.3 g, yield: 59.8%). ESI [M+H] + =330.2

[0407] Ethyl 1-((S)-1-((R)-1-(4-methoxyphenyl)ethyl)-5-oxopyrrolidin-3-yl)cyclopropane-1-carboxylate (049-D) and (049-D') To a solution of compound 49-C (1.3 g, 4 mmol) in EA (30 mL) was added Pd / C (200 mg) and the mixture was stirred at room temperature overnight under H2 atmosphere. The mixture was filtered through a celite pad, concentrated and purified by chromatography on silica gel eluted with PE:EA (10:1 to 3:1) to give the desired product as a mixture of two isomers (1 g, yield: 75%). ESI: [M+H] + =332.2. The two diastereomers were further separated by preparative HPLC.

[0408] Ethyl (S)-1-(5-oxopyrrolidin-3-yl)cyclopropane-1-carboxylate (49-E) A solution of compounds 49-D and 49-D' (1:1, 470 mg, 1.42 mmol) in TFA (4 mL) was stirred at reflux for 3 h. The mixture was concentrated and purified by chromatography on silica gel eluting with DCM:MeOH (10:1 to 3:1) to give the desired product (250 mg, yield: 89.4%). ESI: [M+H] + =198.1

[0409] Ethyl (R)-1-(1-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-5-oxopyrrolidin-3-yl)cyclopropane-1-carboxylate (49-F) Compound 49-F was synthesized using the procedure described above. ESI: [M+H] + =518.2

[0410] (R)-1-(1-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a] [1,4]Diazepin-7-yl)-5-oxopyrrolidin-3-yl)cyclopropanecarboxylic acid (49-G) To a solution of compound 49-F (100 mg, 0.19 mmol) in THF (5 mL) was added a solution of LiOH (46 mg, 1.9 mmol) in water (5 mL), and the mixture was stirred at 40° C. overnight. The reaction mixture was adjusted to pH=2 with 6M HCl and extracted with DCM / MeOH (10 / 1, 20 mL*3). The combined organic layers were washed with brine and dried over Na2SO4. The organic solution was filtered and concentrated in vacuo to give the desired product (90 mg, yield: 94.7%). ESI: [M+H] + =490.2

[0411] tert-Butyl (S)-(1-(1-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-5-oxopyrrolidin-3-yl)cyclopropyl)carbamate (49-H) To a solution of compound 49-G (90 mg, 0.18 mmol) in toluene (20 mL), TEA (37 mg, 0.39 mmol), DPPA (55 mg, 0.2 mmol) were added and stirred at reflux for 1.5 h. To the above solution, t-BuOH (5 mL) was added and stirred at 100° C. overnight. The reaction mixture was cooled to room temperature and concentrated to dryness. The residue was purified by preparative TLC (DCM:MeOH=20:1) to give the desired compound (20 mg, yield: 19%). ESI: [M+H] + =561.3

[0412] (S)-4-(7-(4-(1-aminocyclopropyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (49) To a solution of compound 49-G (20 mg, 0.035 mmol) in MeOH (5 mL), HCl-dioxane (4 M, 3 mL) was added and the resulting solution was stirred at room temperature for 2 h. The mixture was concentrated and purified by preparative HPLC to give the desired compound (11 mg, yield: 68.5%). ESI: [M+H] + = 461.5; 1 H NMR (400 MHz, CD3OD): δ 8.12 (d, J = 2.4 Hz, 1H), 8.03 (s, 1H), 7.84 (dd, J = 8.9, 2.4 Hz, 1H), 7.78 - 7.69 (m, 7H), 7.29 (d, J = 1.7 Hz, 1H), 5.38 (s, 2H), 4.08 (dd, J = 9.8, 8.4 Hz, 1H), 3.81 - 3.69 (m, 1H), 3.04 - 2.93 (m, 1H), 2.80 (dd, J = 17.0, 8.8 Hz, 1H), 2.57 (dd, J = 17.0, 9.9 Hz, 1H), 1.10 - 1.02 (m, 4H).

[0413] Using the procedures described above, the examples in Table 7 below were synthesized.

[0414] [Table 7]

[0415] [ka]

[0416] (S)-4-(7-(4-(2-aminopropan-2-yl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (54) To a solution of compound 52 (100 mg, 0.21 mmol) in 10 mL of MeOH, (HCHO) n (63mg 2.1mmol) was added at 0°C and then stirred at room temperature for 30 min. To the above mixture, NaBH3CN (26mg, 0.42mmol) was added at room temperature and stirred for 1 h. The reaction mixture was concentrated and purified by preparative HPLC to give the desired product (50mg, yield: 50%) as a white solid. ESI: [M+H] + = 491.2; 1 H NMR (400 MHz, DMSO-d6): δ 8.00 - 7.89 (m, 2H), 7.81 (d, J = 1.4 Hz, 1H), 7.79 - 7.67 (m, 5H), 7.58 (d, J = 8.7 Hz, 1H), 7.23 (d, J = 1.4 Hz, 1H), 7.04 (d, J = 2.0 Hz, 1H), 5.24 (s, 2H), 3.93 - 3.68 (m, 2H), 2.87 - 2.70 (m, 1H), 2.56 (dd, J = 17.5, 9.3 Hz, 3H), 2.16 (s, 6H), 0.93 (s, 3H), 0.87 (s, 3H).

[0417] [ka]

[0418] 5-Hydroxy-1-(4-methoxybenzyl)-1,5-dihydro-2H-pyrrol-2-one (55-A) To a solution of 1-(4-methoxybenzyl)-1H-pyrrole-2,5-dione (19 g, 87 mmol) in DCM / MeOH (5:1) (20 mL) was added NaBH4 (3.97 g, 104 mmol) and CeCl3.7H2O (39 g, 104 mmol) at 0 °C. The resulting mixture was stirred for 1 h before being poured into ice-cold HCl (1M in water, 20 mL) and extracted with EA (300 mL*2). The combined organic layers were combined, washed with aq. NaHCO3, brine, dried over Na2SO4, filtered and concentrated to dryness. The residue was purified by chromatography on silica gel eluting with PE / EA (10 / 1 to 1 / 1) to give the desired compound as a white solid (13 g, yield: 68%). ESI: [M+H] + =220.1

[0419] 5-((tert-butyldiphenylsilyl)oxy)-1-(4-methoxybenzyl)-1,5-dihydro-2H-pyrrol-2-one (55-B) To a solution of compound 55-A (13 g, 86 mmol) in DCM (200 mL) was slowly added TBDPSCl (53.8 g, 95 mmol) and imidazole (8.8 g, 129 mmol) at 0° C., and the resulting solution was stirred at 20° C. for 1 h. The mixture was quenched with water and extracted with DCM three times. The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by silica gel chromatography eluting with PE / EA (10 / 1 to 3 / 1) to give the desired compound (26 g, yield: 65%) as a colorless oil. ESI: [M+H] + =458.2

[0420] 5-((tert-butyldiphenylsilyl)oxy)-1-(4-methoxybenzyl)-4-(2-nitropropan-2-yl)pyrrolidin-2-one (55-C) To a solution of compound 52-B (2.0 g, 4.4 mmol) in 2-nitropropane (50 mL), DBU (1.3 g, 8.8 mmol) was added and the resulting solution was stirred at room temperature for 48 h. The mixture was concentrated and purified by chromatography on silica gel eluted with PE / EA (10 / 1 to 1 / 1) to give the desired compound (980 mg, yield: 42%). ESI: [M+H] + =547.3

[0421] 5-Hydroxy-1-(4-methoxybenzyl)-4-(2-nitropropan-2-yl)pyrrolidin-2-one (55-D) To a solution of compound 55-C (980 mg, 0.18 mmol) in THF (10 mL) was added TBAF in THF (1 M, 4 mL) at room temperature, and the resulting solution was stirred at room temperature for 1 h. The reaction mixture was diluted with NH4Cl solution and extracted with DCM (20 mL*2). The combined organic layers were dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by silica gel chromatography eluting with PE / EA (10 / 1 to 1 / 1) to give the desired compound as a yellow oil (500 mg, yield: 89%). ESI: [M+H] + =309.1

[0422] 1-(4-Methoxybenzyl)-4-(2-nitropropan-2-yl)pyrrolidin-2-one (55-E) To a solution of compound 52-D (480 mg, 1.56 mmol) in DCM (30 mL) was added BF3.Et2O (444 mg, 3.12 mmol) and triethylsilane (364 mg, 3.12 mmol) at 0 °C under N2 atmosphere, and the resulting solution was stirred at 20 °C for 8 h. The reaction was quenched with H2O and the mixture was extracted with EA (20 mL * 2). The combined organic layers were dried over Na2SO4, filtered and concentrated to dryness. The residue was purified by chromatography on silica gel eluting with PE / EA (10 / 1 to 1 / 1) to give the desired product as a yellow oil (300 mg, yield: 66%). ESI: [M+H] + =293.1

[0423] 4-(2-aminopropan-2-yl)-1-(4-methoxybenzyl)pyrrolidin-2-one (55-F) To a suspension of compound 55-E (600 mg, 2.1 mmol) in 10 mL of HO and 10 mL of EtOH, Fe (1.2 g, 21 mmol) and NHCl (535 mg, 10.5 mmol) were added. The reaction mixture was then stirred at 80° C. for 2 h, cooled to room temperature, and filtered. The filtrate was concentrated, and the residue was purified by chromatography on silica gel to give the desired compound as a yellow solid (440 mg, yield: 80%). ESI: [M+H] + = 263.2

[0424] 4-(2-(azetidin-1-yl)propan-2-yl)-1-(4-methoxybenzyl)pyrrolidin-2-one (55-G) A mixture of compound 55-F (440 mg, 1.5 mmol), 1,3-dibromopropane (370 mg, 1.8 mmol) and K2CO3 (620 mg, 4.5 mmol) in 20 mL of CH3CN was stirred at 60°C overnight. The reaction mixture was cooled to room temperature, filtered and then concentrated to dryness. The residue (450 mg, crude) was used directly in the next step. ESI: [M+H] + = 303.2

[0425] 4-(2-(azetidin-1-yl)propan-2-yl)pyrrolidin-2-one (55-H) A mixture of compound 55-G (450 mg, crude) in TFA (10 mL) was placed in a microwave reactor at 140° C. for 2 h. The reaction mixture was concentrated to dryness under reduced pressure, and the residue was washed with Et2O (10 mL*3) to give the desired product as a yellow solid (270 mg, yield: 99%). ESI: [M+H] + = 183.1

[0426] 4-(7-(4-(2-(azetidin-1-yl)propan-2-yl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (55) Using the procedure described above, compound 52 was synthesized as a white solid (2 mg, yield: 2%). ESI: [M+H] + = 503.2; 1 H NMR (400 MHz, CD3OD): δ 7.96 (d, J = 2.2 Hz, 1H), 7.77 (dd, J = 8.8, 2.2 Hz, 1H), 7.74-7.43 (m, 7H), 7.18 (d, J = 8.7 Hz, 1H), 6.89 (d, J = 8.7 Hz, 1H), 5.12 (s, 2H), 3.92-3.81 (m, 2H), 3.53 (t, J = 6.7 Hz, 5H), 3.35 (s, 1H), 2.58 (s, 2H), 2.14 (dd, J = 14.5, 7.2 Hz, 2H), 1.09 (s, 6H).

[0427] [ka]

[0428] ((Benzyloxy)carbonyl)-L-asparagine (056-A) To a 0°C solution of L-asparagine (11.2 g, 91.7 mmol) in 1,4-dioxane (112 mL), Na2CO3 (2N, 200 mL) was added, followed by dropwise addition of CbzCl (16.4 g, 96.3 mmol). The resulting mixture was stirred at room temperature for 18 h and then washed with MTBE. The aqueous layer was acidified with HCl (4N) to adjust the pH to <1, resulting in the precipitation of a white solid, which was filtered off and dried in vacuum to give the desired compound (19 g, yield: 77.9%) as a white solid. ESI: [M+H] + = 267.1

[0429] (S)-3-((benzyloxy)carbonyl)-2-oxoimidazolidine-4-carboxylic acid (56-B) To a solution of NaOH (2.98 g, 74.4 mmol) in H2O (60 mL) at 0° C., Br2 (3.65 g, 24.3 mmol) was added slowly. Compound 56-A (6.0 g, 22.5 mmol) was added to the above solution at 0° C., and the resulting mixture became clear and colorless. The resulting solution was stirred at 55° C. for 3 h and cooled to room temperature. The solution was washed with MTBE, and then acidified with HCl (4N) to adjust pH<1. The white solid was filtered and dried in vacuum to give the desired compound (4 g, yield: 66.7%). ESI: [M+H] + = 265.1

[0430] (S)-1-Benzyl-5-methyl 2-oxoimidazolidine-1,5-dicarboxylate (56-C) To a 0°C solution of compound 56-B (2.0 g, 7.55 mmol) in MeOH (20 mL) was added SOCl2 (2.7 g, 22.65 mmol) slowly. The resulting mixture was stirred at 25°C for 3 h. The mixture was quenched with saturated aqueous NaHCO3, then extracted with EA three times. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with PE / EA (10 / 1 to 1 / 2) to give the desired compound as a white solid (1.7 g, yield: 81%). ESI: [M+H] + = 279.1

[0431] (S)-3-Benzyl-1-tert-butyl-4-methyl-2-oxoimidazolidine-1,3,4-tricarboxylate (56-D) To a 0°C solution of compound 56-C (0.23 g, 0.82 mmol) and DMAP (104 mg, 0.82 mmol) in CH3CN (20 mL) was added Boc2O (0.36 g, 1.65 mmol) slowly. The resulting mixture was stirred at 50°C for 3 h. After consumption of the starting material, water was added and the mixture was extracted with EA three times. The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated to dryness under reduced pressure. The residue was purified by chromatography on silica gel eluted with PE / EA (10 / 1 to 1 / 5) to give the desired product as a white solid (0.26 g, yield: 83.6%). ESI: [M+H] + = 379.1

[0432] (S)-1-tert-Butyl 4-methyl-2-oxoimidazolidine-1,4-dicarboxylate (56-E) To a solution of compound 56-D (0.26 g, 0.69 mmol) in MeOH (10 mL) was added Pd / C (10%, 50 mg). The resulting mixture was stirred at 50 °C for 16 h under 50 psi H2 atmosphere. After consumption of the starting material, the mixture was cooled and filtered through a celite pad, and the filtrate was concentrated under reduced pressure to give the desired product as a solid (0.16 g, yield: 94.1%). ESI: [M+H] + = 245.1

[0433] (S)-1-tert-butyl 4-methyl 3-methyl-2-oxoimidazolidine-1,4-dicarboxylate (56-F) To a 0°C solution of compound 56-E (0.16 g, 0.65 mmol) in anhydrous THF (10 mL), NaH (60%, 39 mg, 0.98 mmol) was carefully added. The resulting mixture was stirred at 0°C for 0.5 h. To the above solution, MeI (139 mg, 0.98 mmol) was added dropwise, and the resulting mixture was stirred at room temperature for 2 h. After the starting material was consumed, the mixture was quenched with saturated NH4Cl and extracted with EA three times. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and concentrated to dryness under reduced pressure. The residue was purified by silica gel chromatography eluting with PE / EA (10 / 1 to 1 / 5) to give the desired compound (0.12 g, yield: 75%). ESI: [M+H] + = 259.1

[0434] (S)-Methyl 3-methyl-2-oxoimidazolidine-4-carboxylate (56-G) To a 0° C. solution of compound 56-G (0.5 g, 2 mmol) in MeOH (10 mL) was added HCl in 1,4-dioxane (4N, 10 mL). The resulting mixture was stirred at 20° C. for 6 h. The mixture was concentrated in vacuo to give the desired product as a yellow solid (0.28 g, yield: 88%). ESI: [M+H] + = 159.1

[0435] (S)-Methyl 1-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo [1,2-a][1,4]diazepin-7-yl)-3-methyl-2-oxoimidazolidine-4-carboxylate (56-H) To a solution of Int-A (120 mg, 0.3 mmol) in toluene (10 mL) was added 56-G (48 mg, 0.3 mmol), Pd(OAc)2 (7 mg, 0.03 mmol), Xantphos (35 mg, 0.06 mmol) and Cs2CO3 (293 mg, 0.9 mmol). The reaction was purged with argon gas three times and stirred at 110 °C for 18 h. The reaction mixture was cooled to room temperature, diluted with DCM (30 mL) and filtered. The filtrate was concentrated to dryness and purified by flash chromatography eluting with DCM / MeOH (100 / 1 to 20 / 1) to give the desired compound (18 mg, yield: 12.5%). ESI: [M+H] + = 479.2

[0436] (S)-4-(7-(4-(hydroxymethyl)-3-methyl-2-oxoimidazolidin-1-yl)-9H- Benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (56-I) To a 0°C solution of compound 56-H (18 mg, 0.03 mmol) in EtOH (10 mL) was added NaBH4 (6 mg, 0.15 mmol). The reaction was stirred at 20°C for 2 h. After consumption of the starting material, the mixture was quenched with aqueous NH4Cl and extracted with EA. The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated to dryness in vacuo. The residue was purified by flash chromatography eluting with DCM / MeOH (100 / 1 to 20 / 1) to give the desired compound (15 mg, yield: 95%). ESI: [M+H] + = 451.2

[0437] (R)-4-(7-(4-((dimethylamino)methyl)-3-methyl-2-oxoimidazolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (56) Using the procedure described above, compound 56 was synthesized. ESI: [M+H] + = 478.2; 1H-NMR (400 MHz, CD3OD): δ 8.03 (m, 2H), 7.74 (m, 8H), 7.30 (s, 1H), 5.35 (s, 2H), 4.26 (t, J = 9.2 Hz, 1H), 4.21 - 4.10 (m, 1H), 3.96 - 3.86 (m, 1H), 3.60 (m, 1H), 3.49 - 3.38 (m, 1H), 3.02 (s, 6H), 2.96 (s, 3H).

[0438] [ka]

[0439] 4-(7-(3-(2-hydroxyethyl)-2-oxoimidazolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (57-A) Compound 57-A was synthesized using the procedure described above. ESI: [M+H] + = 451.2

[0440] 4-(7-(3-(2-(dimethylamino)ethyl)-2-oxoimidazolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (57) Using the procedure described above, compound 57 was synthesized. ESI: [M+H] + = 478.2; 1H NMR (400 MHz, CD3OD): δ 8.05 (d, J = 2.1 Hz, 1H), 8.00 (d, J = 2.5 Hz, 1H), 7.84 (dd, J = 9.0, 2.5 Hz, 1H), 7.79 (d, J = 1.8 Hz, 1H), 7.78 - 7.70 (m, 6H), 7.70 (s, 1H), 7.31 (d, J = 1.8 Hz, 1H), 5.37 (s, 2H), 4.01 (dd, J = 9.2, 6.8 Hz, 2H), 3.73 - 3.69 (m, 2H), 3.66 (dd, J = 9.1, 6.9 Hz, 2H), 3.44 - 3.39 (m, 3H), 2.99 (s, 6H).

[0441] [ka]

[0442] tert-Butyl 3-(2-oxoimidazolidin-1-yl)azetidine-1-carboxylate (58-A) To a solution of tert-butyl 3-aminoazetidine-1-carboxylate (1 g, 5.8 mmol) and TEA (0.81 mL, 5.8 mmol) in dioxane (30 mL) was added 2-chloroethyl isocyanate (0.54 mL, 6.38 mmol) in portions over 3 min. The mixture was stirred at ambient temperature for 18 h. Sodium hydride (60% dispersion in mineral oil, 350 mg) was then added to the above solution. The resulting reaction mixture was stirred at ambient temperature for 48 h and quenched by slow addition of water (5 mL). The mixture was concentrated under reduced pressure to 30 mL, and the resulting solution was directly purified by chromatography on silica gel (DCM:MeOH=10:1) to give the desired product (600 mg, yield: 43%). ESI: [M+H] + =242.1

[0443] tert-Butyl 3-(3-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo [1,2-a][1,4]diazepin-7-yl)-2-oxoimidazolidin-1-yl)azetidine-1-carboxylate (58-B) Using the procedure described above, compound 052 was synthesized. ESI: [M+H] + =562.2

[0444] 4-(7-(3-(azetidin-3-yl)-2-oxoimidazolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (58-C) To a solution of compound 58-B (30 mg, 0.05 mmol) in MeOH (5 mL), HCl-dioxane (4N, 5 mL) was added and the resulting solution was stirred at 25° C. overnight. After the reaction was completed, the mixture was concentrated and redissolved in MeOH. The solution was neutralized with basic resin and concentrated to give the desired product (20 mg, yield: 81%). ESI: [M+H] + =462.2

[0445] 4-(7-(3-(1-methylazetidin-3-yl)-2-oxoimidazolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (58) Using the procedure described above, compound 58 was synthesized. ESI: [M+H] + = 476.2; 1 H-NMR (400 MHz, CD3OD): δ 8.06 (d, J = 2.0 Hz, 1H), 7.98 (s, 1H), 7.79 (d, J = 1.1 Hz, 1H), 7.74-7.69 (m, 7H), 7.32 (d, J = 1.7 Hz, 1H), 5.38 (s, 2H), 4.69 (dt, J = 15.6, 7.7 Hz, 2H), 4.54-4.52 (s, 2H), 4.29 (s, 1H), 4.01 (t, J = 7.9 Hz, 2H), 3.76-3.66 (m, 2H), 3.05 (s, 3H).

[0446] [ka]

[0447] Compound 59 was synthesized using the procedure described above. ESI: [M+H] + =504.2; 1 H NMR (400 MHz, CD3OD): δ 7.92 (s, 1H), 7.91(s, 1H), 7.79 (dd, J = 8.9, 2.6 Hz, 1H), 7.74 (d, J = 8.6 Hz, 2H), 7.71 - 7.63 (m, 4H), 7.57 (s, 1H), 7.22 (d, J = 1.7 Hz, 1H), 5.30 (s, 2H), 4.60 - 4.46 (m, 1H), 3.96 (dt, J = 15.1, 6.8 Hz, 3H), 3.78 - 3.62 (m, 2H), 2.94(s, 6H), 2.93 - 2.75 (m, 2H), 2.73 - 2.56 (m, 2H).

[0448] [ka]

[0449] (R)-4-Benzyl 1-tert-butyl 2-(hydroxymethyl)piperazine-1,4-dicarboxylate (60-A) To a stirred solution of tert-butyl (R)-2-(hydroxymethyl)piperazine-1-carboxylate (500 mg, 2.31 mmol) and triethylamine (707 mg, 3.93 mmol) in DCM (20 mL) was added CbzCl (787 mg, 4.62 mmol) dropwise at 0° C. The mixture was stirred at room temperature overnight, then washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel chromatography eluting with PE / EA (10 / 1 to 3 / 1) to give the desired compound (660 mg, yield: 82%). ESI: [M+H] + =351.2

[0450] (S)-4-Benzyl 1-tert-butyl 2-((1,3-dioxoisoindolin-2-yl)methyl)piperazine-1,4-dicarboxylate (60-B) To a stirred mixture of compound 60-A (660 mg, 1.88 mmol), PPh3 (1.5 g, 5.64 mmol), isoindoline-1,3-dione (550 mg, 3.76 mmol) in anhydrous THF (20 mL), DEAD (980 mg, 5.64 mmol) was added dropwise at 0 °C. The mixture was stirred at room temperature for 1 h and then concentrated in vacuo. The resulting residue was purified by silica gel chromatography eluting with PE / EA (100 / 1 to 10 / 1) to give the desired product as a white solid (900 mg, yield: 99%). ESI: [M+H] + =480.

[0451] (S)-4-Benzyl 1-tert-butyl 2-(aminomethyl)piperazine-1,4-dicarboxylate (60-C) To a stirred mixture of compound 60-B (900 mg, 1.88 mmol) in ethanol (20 mL) was added methanamine (580 mg, 18.8 mmol). The mixture was stirred at 90° C. for 3 h. The reaction mixture was concentrated to dryness, dissolved in ether, and filtered. The filtrate was concentrated to give the desired compound as a white solid (650 mg, yield: 99%, crude). ESI: [M+H] + =350.2

[0452] Benzyl (S)-3-(aminomethyl)piperazine-1-carboxylate (60-D) To a stirred mixture of compound 60-C (650 mg, 1.88 mmol) in MeOH (20 mL) was added hydrochloric acid (2 mL, 24.0 mmol). The mixture was stirred at room temperature for 3 h and then concentrated to give the desired product as the hydrochloride salt as a white solid (470 mg, yield: 78%, crude). ESI: [M+H] + = 250.2

[0453] (S)-Benzyl 3-oxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate (60-E) To a stirred mixture of compound 60-D (470 mg, 1.47 mmol), TEA (575 mg, 5.64 mmol) in DCM (20 mL) was added CDI (550 mg, 3.39 mmol). The mixture was stirred at room temperature for 3 h and then concentrated. The residue was purified by silica gel chromatography eluting with DCM / MeOH (100 / 1 to 10 / 1) to give the desired compound as a white solid (250 mg, yield: 62%). ESI: [M+H] + = 276.1

[0454] (S)-Benzyl-2-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo [1,2-a][1,4]diazepin-7-yl)-3-oxohexahydroimidazo[1,5-a]pyrazine-7(1H)-carboxylate (60-F) Compound 60-F was synthesized using the procedure described above. ESI: [M+H] + = 596.2

[0455] (R)-4-(7-(3-oxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (60-G) To a stirred solution of compound 60-F (170 mg, 0.28 mmol) in MeOH (20 mL) was added Pd / C (30 mg). The mixture was stirred at room temperature for 3 days under hydrogen atmosphere. The reaction mixture was filtered and the filtrate was concentrated to give the desired product (130 mg, yield: 99%, crude). ESI: [M+H] + = 462.2

[0456] (R)-4-(7-(7-methyl-3-oxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (60) To a stirred mixture of compound 60-G (130 mg, 0.28 mmol) in methanol (10 mL) was added paraformaldehyde (100 mg, 3.3 mmol) and sodium triacetoxyborohydride (190 mg, 0.89 mmol). The mixture was stirred at room temperature for 1 h and then diluted with NaHCO3 before being extracted three times with DCM. The combined organic layers were concentrated and the residue was purified by preparative HPLC to give the desired compound as a white solid (10 mg). ESI: [M+H] + =476.2; 1 H NMR (400 MHz, CD3OD): δ 8.04 (d, J = 2.0 Hz, 1H), 8.00 (d, J = 2.5 Hz, 1H), 7.82 (dt, J = 5.4, 2.7 Hz, 1H), 7.79 (t, J = 2.6 Hz, 1H), 7.77 -7.73 (m, 2H), 7.73 - 7.68 (m, 4H), 7.31 (d, J = 1.8 Hz, 1H), 5.36 (s, 2H), 4.27 - 4.10 (m, 3H), 3.75 (dd, J = 9.1, 3.4 Hz, 1H), 3.63 (t, J = 13.1 Hz, 1H), 3.53 (d, J = 12.0 Hz, 1H), 3.38 (m,1H), 3.16 - 3.00 (m, 2H), 2.96 (s, 3H).

[0457] [ka]

[0458] 1-((R)-2,2-Dimethyl-1,3-dioxolan-4-yl)ethan-1-ol (61-A) To a stirred mixture of (R)-2,2-dimethyl-1,3-dioxolane-4-carbaldehyde (12.0 g, 92.3 mmol) in tetrahydrofuran (200 mL), methylmagnesium bromide in diethyl ether (46 mL, 138 mmol, 3N) was added dropwise under ice cooling, and the resulting mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated to give the desired product (12 g, yield: 88%, crude).

[0459] 2-((S)-1-((S)-2,2-dimethyl-1,3-dioxolan-4-yl)ethyl)isoindoline-1,3-dione (61-B) and 2-((R)-1-((S)-2,2-dimethyl-1,3-dioxolan-4-yl)ethyl)isoindoline-1,3-dione (61-B') To a stirred mixture of isoindoline-1,3-dione (14.5 g, 97.0 mmol), compound 58-A (12 g, 82.0 mmol), PPh3 (25.4 g, 97.0 mmol) in THF (200 mL), DEAD (19.6 g, 97.0 mmol) was added dropwise at 0° C. The mixture was stirred at room temperature for 24 h. The reaction mixture was then diluted with water and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, concentrated, and purified by silica gel chromatography (PE:EA=1:1) to give compounds 61-B′ (4 g, yield: 18%) and 61-B (1 g, yield: 4.5%).

[0460] 2-((R)-1-((S)-2,2-dimethyl-1,3-dioxolan-4-yl)ethyl)isoindoline-1,3-dione (61-B') 1H NMR (400 MHz, CDCl3): δ 7.83 (m, 2H), 7.73 - 7.68 (m, 2H), 4.76 - 4.67 (m, 1H), 4.40 - 4.28 (m, 1H), 4.14 (dd, J = 8.6, 6.2 Hz, 1H), 3.83 (dd, J = 8.6, 5.1 Hz, 1H), 1.40 (t, J = 5.7 Hz, 3H), 1.37 (s, 4H), 1.29 (s, 3H).

[0461] 2-((S)-1-((S)-2,2-dimethyl-1,3-dioxolan-4-yl)ethyl)isoindoline-1,3-dione (61-B). 1 H NMR (400 MHz, CDCl3): δ 7.87 - 7.80 (m, 2H), 7.76 - 7.69 (m, 2H), 4.72 (dt, J = 9.2, 6.0 Hz, 1H), 4.33 (dq, J = 9.2, 6.9 Hz, 1H), 3.96 (dd, J = 8.4, 6.2 Hz, 1H), 3.68 (dt, J = 13.7, 6.9 Hz, 1H), 1.56 (d, J = 6.9 Hz, 3H), 1.44 (s, 3H), 1.38 (d, J = 6.4 Hz, 3H).

[0462] 2-((2S,3S)-3,4-dihydroxybutan-2-yl)isoindoline-1,3-dione (061-C) To a stirred mixture of compound 61-B (1 g, 3.63 mmol) in methanol (20 mL) was added p-toluenesulfonic acid (1.60 g, 9.08 mmol). The mixture was stirred at 60° C. for 4 h and then cooled to room temperature. The reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, concentrated and purified by silica gel chromatography (PE:EA=1:10) to give the desired product (850 mg, yield: 99%). ESI: [M+H] + =236.1

[0463] 2-((2S,3S)-3-Hydroxy-4-(trityloxy)butan-2-yl)isoindoline-1,3-dione (61-D) To a stirred mixture of compound 61-C, (chloromethanetriyl)tribenzene (1.27 g, 4.59 mmol), TEA (780 mg, 7.64 mmol) in DCM (30 mL) was added DMAP (24 mg, 0.20 mmol). The mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with EA and washed with water. The organic layer was dried over sodium sulfate, concentrated and purified by silica gel chromatography (PE:EA=1:1) to give the desired product (1.5 g, yield: 87%). ESI: [M+H] + =478.2

[0464] (2S,3S)-3-Amino-1-(trityloxy)butan-2-ol (61-E) To a stirred mixture of compound 61-D (1.5 g, 3.14 mmol) in ethanol (50 mL) was added hydrazine hydrate (1.5 g, 31.4 mmol). The mixture was stirred at 80° C. for 1 h and cooled to room temperature. The reaction mixture was filtered, and the filtrate was dried over sodium sulfate, filtered, and concentrated to give the desired product (1 g, yield: 92%). ESI: [M+H] + =348.2

[0465] (4S,5S)-4-Methyl-5-((trityloxy)methyl)oxazolidin-2-one (61-F) To a stirred mixture of compound 61-E (1.0 g, 2.88 mmol) in tetrahydrofuran (30 mL) was added N,N'-carbonyldiimidazole (740 mg, 4.32 mmol). The mixture was stirred at 65°C for 1 h and then concentrated to dryness. The residue was purified by silica gel chromatography (PE:EA=1:10) to give the desired compound (700 mg, yield: 65%). ESI: [M+H] + =374.2

[0466] 4-(7-((4S,5S)-4-Methyl-2-oxo-5-((trityloxy)methyl)oxazolidin-3-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (61-G) Using the procedure described above, compound 061-F was synthesized. ESI: [M+H] + =694.3

[0467] 4-(7-((4S,5S)-5-(hydroxymethyl)-4-methyl-2-oxooxazolidin-3-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (61-H) To a stirred solution of compound 61-G (200 mg, 0.29 mmol) in DCM (10 mL) was added TFA (160 mg, 1.44 mmol). The mixture was stirred at room temperature for 1 h and then diluted with DCM. The organic solution was washed with water, dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica gel chromatography (DCM:CH3OH=10:1) to give the desired compound (100 mg, yield: 77%). ESI: [M+H] + =452.2

[0468] 4-(7-((4S,5R)-5-((dimethylamino)methyl)-4-methyl-2-oxooxazolidin-3-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (61) Using the procedure described above, compound 61 was synthesized. ESI: [M+H] + = 479.2; 1H NMR (400 MHz, CD3OD): δ 8.01(d, J = 2.2 Hz, 1H), 7.99 (d, J = 2.0 Hz, 1H), 7.78 - 7.64 (m, 8H), 7.28 (d, J = 1.7 Hz, 1H), 5.38 (s, 2H), 5.29 - 5.20 (m, 1H), 4.97 - 4.89 (m, 1H), 3.85 - 3.74 (m, 1H), 3.60 (d, J = 13.8 Hz, 1H), 3.04 (s, 6H), 1.28 (d, J = 6.4 Hz, 3H).

[0469] [ka]

[0470] N-Methoxy-1-(4-methoxybenzyl)-N-methyl-5-oxopyrrolidine-3-carboxamide (62-A) To a solution of 1-(4-methoxybenzyl)-5-oxopyrrolidine-3-carboxylic acid (15.2 g, 60.8 mmol), N,O-dimethylhydroxylamine (6 g, 60.8 mmol), DIPEA (23.6 g, 182.4 mmol) in dry DMF (100 mL) was added HATU (23.1 g, 60.8 mmol) at 0° C. The resulting mixture was stirred at room temperature for 2 h and diluted with EA. The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated to dryness. The crude product was purified by silica gel chromatography to give the desired product as an oil (16 g, yield: 90%). ESI: [M+H] + = 293.1

[0471] 4-Acetyl-1-(4-methoxybenzyl)pyrrolidin-2-one (62-B) To a solution of compound 62-A (11.7 g, 40 mmol) in dry THF (120 mL) was added MeMgBr (60 mmol, 20 mL) at 0 °C. The resulting mixture was stirred at room temperature for 3 h before being poured into 1N HCl solution at 0 °C. After extraction with EA, the combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with PE / EA (1 / 1 to 1 / 20) to give the desired product as a yellow oil (7.3 g, yield: 73.9%). ESI: [M+H] + = 248.1

[0472] (S,E)-N-(1-(1-(4-methoxybenzyl)-5-oxopyrrolidin-3-yl)ethylidene)-2-methylpropane-2-sulfinamide (62-C) To a 0°C solution of compound 62-B (6.3 g, 25.4 mmol) in dry THF (100 mL), (S)-2-methylpropane-2-sulfinamide (4.6 g, 38.1 mmol) and Ti(O-iPr)4 (21.6 g, 76.2 mmol) were added. The resulting mixture was stirred at 20°C for 24 h. To the above solution, saturated aqueous NaHCO3 was added at 0°C to quench the reaction. The resulting mixture was filtered through a celite pad and the filtrate was extracted with EA. The organic phase was washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by chromatography on silica gel eluting with PE / EA (1 / 1 to 1 / 10) to give the desired product (7.7 g, yield: 86.5%). ESI: [M+H] + = 351.2

[0473] (S)-N-(2-(1-(4-methoxybenzyl)-5-oxopyrrolidin-3-yl)pent-4-en-2-yl)ethanesulfinamide (62-D) To a 0°C solution of compound 62-C (6.0 g, 17.1 mmol) in dry THF (50 mL) was added allylmagnesium bromide (25.6 mmol, 25.6 mL). The resulting reaction mixture was stirred at 20°C for 3 h, then quenched by adding 1N NH4Cl at 0°C and extracted with EA three times. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by chromatography on silica gel eluting with PE / EA (1 / 1 to 1:20) to give the desired compound as two isomers as an oil (1 g, P1; 1.8 g P2, yield: 42.5%). ESI: [M+H] + = 393

[0474] (S)-N-(2-(1-(4-methoxybenzyl)-5-oxopyrrolidin-3-yl)pent-4-en-2-yl)ethanesulfinamide (62-E) A -78 °C solution of compound 62-D (1.8 g, 4.5 mmol) in dry MeOH (30 mL) was purged with O3 until the resulting mixture turned blue. After the starting material was consumed, the solution was purged with N2. NaBH4 (684 mg, 18 mmol) was added to the solution and the resulting solution was stirred at 20 °C for 30 min before adding aqueous NH4Cl and extracting with EA. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography eluting with DCM / MeOH (100 / 1 to 20 / 1) to give the desired product as a white solid (810 mg, yield: 44%). ESI: [M+H] + = 397.2

[0475] 4-(1-((S)-tert-butylsulfinyl)-2-methylazetidin-2-yl)-1-(4-methoxybenzyl)pyrrolidin-2-one (62-F) To a solution of compound 62-E (0.7 g, 1.76 mmol) and PPh3 (2.31 g, 8.83 mmol) in dry THF (50 mL) was added DIAD (2.13 g, 10.56 mmol) at 20 °C. The reaction was stirred at 70 °C for 16 h and then concentrated in vacuo. The residue was purified by silica gel chromatography eluting with DCM / MeOH (100 / 1 to 10 / 1) to give the desired product as a yellow solid (0.5 g, yield: 75%). ESI: [M+H] + = 379.2

[0476] 1-(4-Methoxybenzyl)-4-(2-methylazetidin-2-yl)pyrrolidin-2-one (62-G) To a solution of compound 62-F (0.5 g, 1.32 mmol) in 1,4-dioxane (10 mL) was added HCl (12N, 2 mL) at 0° C. The reaction was stirred at 20° C. for 2 h and then concentrated in vacuo to give the desired compound as the HCl salt, which was used directly in the next step (0.41 g, yield: 99%). ESI: [M+H] + = 275.2

[0477] tert-Butyl-2-(1-(4-methoxybenzyl)-5-oxopyrrolidin-3-yl)-2-methylazetidine-1-carboxylate (62-H) To a solution of compound 62-G (0.4 g, 1.32 mmol) and TEA (404 mg, 4.0 mmol) in DCM (10 mL) was added Boc2O (0.29 g, 1.32 mmol) slowly at 0 °C. The resulting mixture was stirred at 20 °C for 2 h. The mixture was diluted with water and extracted with EA three times. The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with PE / EA (10 / 1 to 1 / 10) to give the desired product as a white solid (0.33 g, yield: 66.7%). ESI: [M+H] + = 375.2

[0478] tert-Butyl 2-methyl-2-(5-oxopyrrolidin-3-yl)azetidine-1-carboxylate (62-I) To a solution of compound 62-H (0.1 g, 0.27 mmol) in CH3CN (10 mL) was added CAN (740 mg, 1.35 mmol) at 0 °C. The resulting mixture was stirred at 20 °C for 2 h and diluted with water. After extraction with EA three times, the organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with DCM / MeOH (100 / 1 to 10 / 1) to give the desired compound (40 mg, yield: 58%). ESI: [M+H] + = 255.2

[0479] tert-Butyl 2-(1-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo [1,2-a][1,4]diazepin-7-yl)-5-oxopyrrolidin-3-yl)-2-methylazetidine-1-carboxylate (62-J) Compound 62-J was synthesized using the procedure described in General Procedure I. ESI: [M+H] + = 575.3

[0480] 4-(7-(4-(2-methylazetidin-2-yl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (62) To a solution of compound 62-J (28 mg, 0.05 mmol) in anhydrous DCM (10 mL) was added TFA (2 mL). The resulting reaction solution was stirred at room temperature for 3 h before being adjusted to pH>7 by adding saturated aqueous NaHCO3. The mixture was then extracted with DCM; the combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by preparative HPLC to give the desired compound (8 mg, yield: 33.3%). ESI: [M+H] + = 475.2 1H NMR (400 MHz, CD3OD): δ 8.13 (d, 2.4 Hz, 1H), 8.05 (s, 1H), 7.91 (m, 1H), 7.74 (m, 7H), 7.30 (s, 1H), 5.39 (s, 2H), 4.28 - 3.69 (m, 5H), 2.91 (m, 1H), 2.68 - 2.53 (m, 2H), 2.44 (m, 1H), 1.63 (s, 3H).

[0481] [ka]

[0482] Methyl 3-((tert-butoxycarbonyl)amino)oxetane-3-carboxylate (63-A) To a solution of methyl 3-aminooxetane-3-carboxylate (1.7 g, 13 mmol) and di-tert-butyl dicarbonate (3.1 g, 14.3 mmol) in DCM (17 mL) was added TEA (2.63 g, 26 mmol) and the mixture was stirred at 20° C. for 4 h under nitrogen atmosphere before diluting with DCM. The resulting solution was washed with water, brine and concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with PE / EA (10 / 1) to give the desired compound (1.0 g, yield: 33%). ESI: [M+H] + =232.2

[0483] tert-Butyl (3-(hydroxymethyl)oxetan-3-yl)carbamate (63-B) To a solution of compound 63-A (2.0 g, 8.7 mmol) in MeOH (20 mL) was added NaBH4 (1.0 g, 26.1 mmol) and the mixture was stirred at 20° C. overnight under nitrogen atmosphere before the reaction was quenched by adding aqueous NH4Cl. The resulting mixture was extracted three times with EtOAc. The combined organic layers were washed with brine and concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with PE / EA (1 / 1) to give the desired compound (1.7 g, yield: 94%). ESI: [M+H]+ =204.1; 1 H NMR (400 MHz, CDCl3): δ 5.09 (s, 1H), 4.66 (d, J = 6.7 Hz, 2H), 4.54 (d, J = 6.5 Hz, 2H), 4.03 (s, 2H), 1.45 (s, 9H).

[0484] tert-Butyl (3-formyloxetan-3-yl)carbamate (63-C) To a solution of compound 63-B (1.7 g, 8.4 mmol) in DCM (20 mL) was added Dess-Martin periodinane (5.3 g, 12.6 mmol) and the mixture was stirred at 20° C. overnight under nitrogen atmosphere. The mixture was then quenched with aqueous NaHCO3 and Na2SO3 and extracted with DCM. The organic phase was washed with brine and concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with PE / EtOAc (1 / 1) to give the desired compound (1.4 g, yield: 82%). ESI: [M+H] + =202.1

[0485] Ethyl-3-(3-((tert-butoxycarbonyl)amino)oxetan-3-yl)acrylate (63-D) A solution of 63-C (1.4 g, 7.0 mmol) and ethyl 2-(triphenyl-15-phosphanylidene)acetate (3.7 g, 10.5 mmol) in toluene (20 mL) was stirred at 110° C. overnight under nitrogen atmosphere. After cooling to room temperature, the mixture was concentrated under reduced pressure and the residue was purified by silica gel chromatography eluting with PE / EA (5 / 1) to give the desired compound (1.0 g, 53% yield). ESI: [M+H] + =272.1; 1H NMR (400 MHz, CDCl3): δ 7.33 (d, J = 15.7 Hz, 1H), 6.00 (d, J = 15.7 Hz, 1H), 5.29 (s, 1H), 4.77 (d, J = 6.5 Hz, 2H), 4.64 (s, 2H), 4.22 (q, J = 7.1 Hz, 2H), 1.44 (s, 9H), 1.31 (t, J = 7.1 Hz, 3H).

[0486] Ethyl 3-(3-((tert-butoxycarbonyl)amino)oxetan-3-yl)-4-nitrobutanoate (63-E) To a solution of compound 63-D (0.794 g, 2.93 mmol) in nitromethane (8 mL) was added DBU (2.3 g, 14.7 mmol) and the mixture was stirred at 70° C. for 3 h under nitrogen atmosphere. The mixture was diluted with DCM, washed with water, brine and concentrated to dryness under reduced pressure. The residue was purified by silica gel chromatography eluting with PE / EtOAc (3 / 1) to give the desired product (0.86 g, yield: 89%). ESI: [M+H] + =333.2

[0487] tert-Butyl (3-(5-oxopyrrolidin-3-yl)oxetan-3-yl)carbamate (63-F) To a solution of compound 63-E (0.39 g, 1.1 mmol) in MeOH (5 mL) was added Pd / C on carbon 10% (0.05 g, 0.44 mmol) and the mixture was stirred under H2 balloon at 45 °C for 1 h. The mixture was then cooled to room temperature, filtered through celite, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with PE / EtOAc (1 / 1) to give the desired product (0.2 g, yield: 67%). ESI: [M+H] + =257.1

[0488] tert-Butyl (3-(1-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo [1,2-a][1,4]diazepin-7-yl)-5-oxopyrrolidin-3-yl)oxetan-3-yl)carbamate (63-G) Compound 063-G was synthesized using the procedure described in General Procedure I. ESI: [M+H] + =577.2

[0489] 4-(7-(4-(4-(hydroxymethyl)-2-oxooxazolidin-4-yl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (63) To a solution of compound 63-G (3 mg, 0.005 mmol) in DCM (2 mL) was added TFA (0.5 mL), and the mixture was stirred at 20° C. for 1 h, then concentrated under reduced pressure. The residue was purified by preparative HPLC to give the title compound (1.4 mg, yield: 52%). ESI: [M+H] + =521.2; 1 H NMR (400 MHz, CD3OD): δ 8.11-8.06 (m, 2H), 7.98-7.91 (m, 1H), 7.80 (s, 1H), 7.75-7.70 (m, 7H), 7.31 (s, 1H), 5.40 (s, 2H), 4.45 - 4.27 (m, 2H), 4.11-4.01 (m, 1H), 3.97 - 3.83 (m, 1H), 3.67-3.54 (m, 2H), 3.0 -2.90 (m, 1H), 2.68 (dd, J = 9.5, 7.1 Hz, 2H).

[0490] [ka]

[0491] tert-Butyl 3-cyano-3-methylazetidine-1-carboxylate (64-A). To a solution of tert-butyl 3-cyanoazetidine-1-carboxylate (1.5 g, 8.2 mmol) in THF (30 mL) was added LiHMDS (1.0 M solution in THF, 9.0 mL, 9.0 mmol) dropwise over 25 min at -78°C. The yellow solution was stirred at this temperature for 30 min, then iodomethane (0.78 mL, 12.5 mmol) was added slowly. The reaction mixture was stirred at this temperature for an additional 30 min, then warmed to room temperature over 1 h. The reaction mixture was quenched with 10 mL of saturated aqueous NH4Cl, diluted with 10 mL of water, and then extracted twice with about 100 mL of EtOAc. The combined organic layers were washed with 20 mL of water and 20 mL of brine, then dried over sodium sulfate, filtered, and concentrated. The residue was chromatographed on silica gel with EtOAc / Hexanes (gradient: 0-30% EtOAc) to give the desired product (1.2 g, yield: 74.6%). ESI: [M+H] + =197.1

[0492] tert-Butyl 3-(aminomethyl)-3-methylazetidine-1-carboxylate (64-B) To a solution of compound 64-A (1.2 g, 6.12 mmol) in MeOH (20 mL), Raney Ni (3 g) was added and stirred at room temperature overnight under H2. The reaction mixture was then filtered and the filtrate was concentrated to give the desired product (900 mg, yield: 73.5%). ESI: [M+H] + =201.2

[0493] tert-Butyl 3-((2,5-dioxoimidazolidin-1-yl)methyl)-3-methylazetidine-1-carboxylate (064-C) To a solution of compound 64-B (900 mg, 4.5 mmol) in dry DCM (20 mL) was added ethyl 2-isocyanatoacetate (580 mg, 4.5 mmol) at 0° C., and the resulting solution was stirred at room temperature for 2 h before NaH (216 mg, 9 mmol) was added. The reaction mixture was stirred at room temperature for another 1 h, and then MeOH (10 mL) was added. The mixture was concentrated, and the residue was purified by chromatography on silica gel with (MeOH / DCM=1 / 10) to give the desired product (1 g, yield: 78.4%). ESI: [M+H] + =284.2.

[0494] tert-Butyl 3-((3-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo [1,2-a][1,4]diazepin-7-yl)-2,5-dioxoimidazolidin-1-yl)methyl)-3-methylazetidine-1-carboxylate (64-D) The desired compound was synthesized using the procedure described in General Procedure I. ESI: [M+H] + =604.3

[0495] 4-(7-(3-((3-methylazetidin-3-yl)methyl)-2,4-dioxoimidazolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (64) To a solution of compound 064-D (350 mg, 0.57 mmol) in MeOH (15 mL), HCl-dioxane (4 M, 5 mL) was added, and the resulting solution was stirred at room temperature for 1 h and then concentrated. The residue was purified by chromatography on silica gel to give the desired compound (264 mg, yield: 92.1%). ESI: [M+H] + =504.2; 1H NMR (400 MHz, CD3OD): δ 8.07 (d, J = 2.3 Hz, 1H), 8.05 (d, J = 2.3 Hz, 1H), 7.95 (dd, J = 8.9, 2.6 Hz, 1H), 7.80 - 7.69 (m, 7H), 7.30 (d, J = 1.8 Hz, 1H), 5.40 (s, 2H), 4.59 (s, 2H), 4.23 (d, J = 11.5 Hz, 2H), 3.89 - 3.67 (m, 4H), 1.43 (s, 3H).

[0496] 4-(7-(3-((1,3-dimethylazetidin-3-yl)methyl)-2,4-dioxoimidazolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (65) The desired compound was synthesized using the procedure described in General Procedure IV. ESI: [M+H] + =518.2; 1 H NMR (400 MHz, CD3OD): δ 8.06 (s,1H), 8.05(s, 1H), 7.96 (d, J = 8.8 Hz, 1H), 7.78-7.31 (m, 7H), 7.31 (d, J = 1.5 Hz, 1H), 5.39 (s, 2H), 4.59 (s, 2H), 4.50 (d, J = 11.0 Hz, 1H), 4.14 (d, J = 10.8 Hz, 1H), 4.00 (d, J = 10.6 Hz, 1H), 3.85-3.84 (m, 3H), 2.94 (s, 3H), 1.43 (s, 3H).

[0497] Using the procedures described above, the following examples in Table 8 were synthesized.

[0498] [Table 8]

[0499] [ka]

[0500] 4-(7-((diphenylmethylene)amino)-9H-benzo[e]imidazo[2,1-c]pyrrolo [1,2-a][1,4]diazepin-12-yl)benzonitrile (Int-B-1) A mixture of compound Int-A (1 g, 2.5 mmol), diphenylmethanimine (905 mg, 5 mmol), Pd2(dba)3 (566 mg, 0.36 mmol), BINAP (738 mg, 1.25 mmol) and t-BuONa (736 mg, 7.15 mmol) in toluene (20 mL) was stirred at 100 °C for 16 h under N2 atmosphere. The mixture was cooled to room temperature and diluted with DCM (50 mL) and MeOH (5 mL). The organic solution was washed with water (50 mL), brine and dried over Na2SO4. After filtration and concentration, the residue was purified by silica gel chromatography to give the desired compound (1.1 g, yield: 88.1%). ESI: [M+H] + = 502.2

[0501] 4-(7-Amino-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (Int-B) To a solution of compound Int-B-1 (1.1 g, 2.19 mmol) in MeOH (20 mL), aqueous HCl (1 M, 10 mL) was added and the resulting solution was stirred at room temperature for 2-3 h. The solid formed in the reaction was filtered and dried to give the desired product as the HCl salt (700 mg, yield: 94.7%). ESI: [M+H] + = 338.1

[0502] Phenyl(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)carbamate (70-A) To a solution of compound Int-A (100 mg, 0.30 mmol) and DIPEA (0.5 mL) in DCM (5 mL) was added phenyl carbonochloridate (70 mg, 0.44 mmol) at room temperature. The resulting solution was stirred for 2-3 h and then concentrated to give the crude desired compound, which was used directly in the next step. ESI: [M+H] + = 458.2

[0503] 1-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-3-(2,2-dimethoxyethyl)urea (70-B) To a solution of compound 70-A (crude, 0.30 mmol) in DCM (5 mL) was added 2,2-dimethoxyethan-1-amine (32 mg, 0.31 mmol) at room temperature, and the resulting solution was stirred for 16 h. The reaction solution was diluted with DCM (200 mL), washed with H2O (20 mL), brine, and dried over Na2SO4. After filtration and concentration, the residue was purified by chromatography on silica gel to give the desired product as a solid (53 mg, yield: 74.6%). ESI: [M+H] + = 469.2

[0504] 4-(7-(2-oxo-2,3-dihydro-1H-imidazol-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (70-C) To a solution of compound 70-B (104 mg, 0.22 mmol) in dioxane (5 mL), HCl (aq) (1 M, 4 mL) was added and stirred at 85 °C for 2-3 h. After cooling to room temperature, the solid formed in the reaction mixture was filtered and dried to give the desired product (50 mg, yield: 56.2%). ESI: [M+H] + = 405.1

[0505] 4-(7-(3-(2-(dimethylamino)ethyl)-2-oxo-2,3-dihydro-1H-imidazol-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (70) To a solution of compound 70-C (50 mg, 0.12 mmol) in DMF (2 mL) was added NaH (20 mg, 0.5 mmol) at 0° C., and the resulting solution was stirred at room temperature for 30 min before 2-bromo-N,N-dimethylethan-1-amine (28 mg, 0.18 mmol) was added. The solution was then stirred at room temperature for 2 h and then quenched with NH4Cl (20 mL); the resulting mixture was extracted with DCM (20 mL). The organic phase was washed with brine, dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative TLC to give the desired compound as a solid (13 mg, yield: 22.4%). ESI: [M+H] + = 476.2; 1 H NMR (400 MHz, CD3OD): δ 8.11 (d, J = 2.4 Hz, 1H), 8.06 (d, J = 2.0 Hz, 1H), 7.96 (dd, J = 8.8, 2.5 Hz, 1H), 7.82 (d, J = 8.8 Hz, 1H), 7.79 - 7.67 (m, 6H), 7.30 (d, J = 1.8 Hz, 1H), 7.12 (d, J = 3.2 Hz, 1H), 6.82 (d, J = 3.2 Hz, 1H), 4.15 (t, J = 5.8 Hz, 2H), 3.55 (t, J = 5.8 Hz, 2H), 2.99 (s, 6H).

[0506] [ka]

[0507] 4-(7-((2,2-dimethoxyethyl)amino)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (71-A) Using the procedure described above, compound 071-A was synthesized.

[0508] tert-Butyl 3-(3-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-3-(2,2-dimethoxyethyl)ureido)azetidine-1-carboxylate (71-B) To a solution of compound 71-A (59 mg, 0.14 mmol) in CH3CN (10 mL) was added 4-nitrophenyl carbonochloridate (30 mg, 0.15 mmol) slowly at 0° C. The resulting mixture was stirred at 20° C. for 3 h and then concentrated under reduced pressure to give the crude product (82 mg), which was used directly in the next step. ESI: [M+H] + = 591.2 To a solution of the above compound (82 mg, 0.14 mmol) in DMAc (3 mL) at 20° C., tert-butyl 3-aminoazetidine-1-carboxylate (72 mg, 0.42 mmol) was slowly added. The resulting mixture was placed in a microwave reactor at 120° C. for 3 h. The reaction mixture was diluted with DCM, washed with brine, and dried over Na2SO4. After filtration, the organic solution was concentrated in vacuo to give the desired crude compound (87 mg), which was used directly in the next step. ESI: [M+H] + = 624.3

[0509] tert-Butyl-3-(3-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-2-oxo-2,3-dihydro-1H-imidazol-1-yl)azetidine-1-carboxylate (71-C) To a solution of compound 71-B (87 mg, 0.14 mmol) in DCM (20 mL) was added HCl in dioxane (4N, 0.5 mL) slowly at 0° C. The resulting mixture was stirred at 20° C. for 3 h, then quenched with saturated aqueous NaHCO3 to adjust pH>7. The resulting mixture was extracted with DCM three times. The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative TLC to give the desired compound as a yellow solid (10 mg, yield: 12%). ESI: [M+H] + =560.2

[0510] 4-(7-(3-(azetidin-3-yl)-2-oxo-2,3-dihydro-1H-imidazol-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (71-D) To a solution of compound 71-C (10 mg, 0.02 mmol) in DCM (5 mL) was added TFA (1 mL). The reaction solution was stirred at room temperature for 1 h. The reaction was quenched with saturated aqueous NaHCO3 to adjust the pH > 7, and the mixture was extracted three times with DCM. The combined organic layers were washed with brine, dried over Na2SO4, and concentrated in vacuo to give the desired crude product (9 mg). ESI: [M+H] + = 460.2

[0511] 4-(7-(3-(1-methylazetidin-3-yl)-2-oxo-2,3-dihydro-1H-imidazol-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (71) Using the procedure described above, compound 71 was synthesized (2 mg, yield: 22.1%). ESI: [M+H] = 474.2; 1H NMR (400 MHz, CD3OD): δ 8.07 (d, J = 2.4 Hz, 1H), 7.93 (s, 1H), 7.78 - 7.66 (m, 5H), 7.54 (s, 1H), 7.48 (m, 1H), 7.35 (m, 1H), 7.22 (d, J = 1.5 Hz, 1H), 7.08 (d, J = 3.2 Hz, 1H), 6.81 (s, 1H), 5.35 (s, 2H), 5.11 - 4.99 (m, 1H), 4.72 - 4.30 (m, 4H), 3.12 (s, 3H).

[0512] [ka]

[0513] N-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-3-(dimethylamino)cyclopentane-1-carboxamide (72) To a solution of compound 66-A (50 mg, 0.11 mmol) in DCM (5 mL) was added N,N-dimethylpyrrolidin-3-amine (15 mg, 0.13 mmol). The reaction mixture was stirred at room temperature overnight and then diluted with DCM (100 mL). The organic phase was washed with H2O (20 mL), brine, dried over Na2SO4, and filtered. The solution was concentrated and the residue was purified by preparative HPLC to give the desired compound as a white solid (23 mg, yield: 45.1%). ESI: [M+H] + = 478.2; 1H NMR (400 MHz, CD3OD): δ 8.04-8.03 (d, J = 2.2 Hz, 1H), 7.90-7.89 (d, J = 2.0 Hz, 1H), 7.80-7.79 (d, J = 4 Hz, 1H),7.80-7.63 (m, 8H),7.31-7.30 (d, J = 4 Hz, 1H), 5.33 (s, 2H), 4.05-3.94 (m, 2H), 3.82-3.80(m, 1H), 3.66-3.59 (m, 2H), 2.98 (s, 6H), 2.54-2.53 (m, 1H),2.30-2.23(m, 1H).

[0514] Using the procedures described above, the examples in Table 9 below were synthesized.

[0515] [Table 9]

[0516] [ka]

[0517] 4-(7-(2,4-dioxoimidazolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (75-A) To a stirred solution of compound Int-B (300 mg, 0.89 mmol), TEA (180 mg, 1.80 mmol) in DMF (10 mL), 2-chloroacetyl isocyanate (127 mg, 1.07 mmol) was added under ice cooling, and the mixture was stirred at room temperature for 1 h. 1,8-diazabicyclo[5.4.0]undec-7-ene (270 mg, 1.80 mmol) was added, and the resulting solution was stirred at room temperature for 1 h. The mixture was diluted with DCM, washed with water and brine, and dried over Na2SO4. After filtration and concentration, the residue was purified by preparative HPLC to give the desired product as a white solid (250 mg, yield: 52%). ESI: [M+H] + =421.1

[0518] 4-(7-(3-(2-(dimethylamino)ethyl)-2,4-dioxoimidazolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (75) A mixture of compound 75-A (70 mg, 0.13 mmol), cesium carbonate (156 mg, 0.48 mmol) and 2-bromo-N,N-dimethylethan-1-amine (75 mg, 0.48 mmol) in DMF (10 mL) was stirred at 100° C. for 1 h. After cooling, the reaction mixture was filtered. The filtrate was concentrated and the residue was purified by preparative HPLC to give the desired compound as a trifluoroacetate salt as a white solid (25 mg, yield: 32%). ESI: [M+H] + = 492.2; 1 H NMR (400 MHz, CD3OD): δ 8.05 (d, J = 2.4 Hz, 1H), 8.01 (s, 1H), 7.96 (m, 1H), 7.78 - 7.73 (m, 4H), 7.70 (d, J = 8.5 Hz, 2H), 7.66 (s, 1H), 7.28 (s, 1H), 5.38 (s, 2H), 4.56 (s, 2H), 4.05-3.98 (m, 2H), 3.54 - 3.44 (m, 2H), 3.02 (s, 6H).

[0519] [ka]

[0520] General Procedure VII: tert-Butyl 3-((12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo [1,2-a][1,4]diazepin-7-yl)carbamoyl)pyrrolidine-1-carboxylate (76-A) A solution of compound Int-B (100 mg, 0.27 mmol) in DMF (10 mL), DIPEA (1 mL) and HATU (205 mg, 0.54 mmol) was stirred at 0° C. To the above solution, 1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid (116 mg, 0.54 mmol) was added. The resulting reaction solution was stirred at room temperature for 3 h and then quenched by the addition of NaHCO3 solution (10 mL). The mixture was extracted three times with DCM / MeOH (10 mL / 2 mL). The combined organic layers were washed with brine and dried over Na2SO4. After filtration, the solution was concentrated and then purified by preparative TLC (DCM:MeOH=15:1) to give the desired product (103 mg, yield: 71.5%). ESI: [M+H] + = 535.2

[0521] N-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4] Diazepin-7-yl)pyrrolidine-3-carboxamide (76) To a solution of compound 76-A (103 mg, 0.19 mmol) in DCM (8 mL) was added TFA (3 mL) dropwise. The reaction solution was stirred for 2 h, then the reaction was quenched with saturated NaHCO3 (10 mL). The mixture was extracted three times with DCM / MeOH (10 mL / 2 mL). The combined organic layers were washed with brine and dried over Na2SO4. After filtration, the solution was concentrated and purified by preparative TLC (DCM:MeOH=15:1) to give the desired product as a pale yellow solid (76 mg, yield: 90.5%). ESI: [M+H] + = 435.2; 1 H NMR (400 MHz, CD3OD): δ 8.08 (d, J =2.2 Hz, 2H), 7.87-7.67 (m, 8H), 7.34 (d, J =1.4 Hz, 1H), 5.38 (s, 2H), 3.68-3.64 (m, 1H), 3.53-3.35 (m, 4H), 2.48-2.39 (m, 1H), 2.31-2.21 (m, 1H).

[0522] N-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-1-isopropylpyrrolidine-3-carboxamide (77) To a solution of compound 76 (37 mg, 0.09 mmol) in THF (3 mL) was added acetone (3 mL) and NaBH3CN (16.1 mg, 0.26 mmol). The reaction mixture was stirred at room temperature for 3 h and quenched with aqueous NaHCO3. The resulting mixture was extracted three times with DCM. The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by preparative HPLC to give the desired product as a white solid (7.3 mg, yield: 18.7%). ESI: [M+H] + = 477.2; 1 H NMR (400 MHz, CD3OD): δ 8.05 (d, J = 6.2 Hz, 1H), 7.96 (s, 1H), 7.79 - 7.61 (m, 8H), 7.25 (d, J = 1.6 Hz, 1H), 5.32 (s, 2H), 3.95-3.60 (m, 5H), 2.68-2.55 (m, 1H), 2.45-2.35(m, 1H), 2.25-2.15 (m, 1H), 1.40 (d, J = 4.0 Hz, 6H).

[0523] Using the procedures described above, the examples in Table 10 below were synthesized.

[0524] [Table 10]

[0525] [ka]

[0526] 4-(7-hydrazinyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile hydrochloride (79-A) To a mixture of compound 79-A (150 mg, 0.45 mmol) in HCl (2 mL), sodium nitrite (30 mg, 0.45 mmol) was added under ice-cooling, the resulting mixture was stirred for 40 min, followed by the addition of SnCl2 (300 mg, 1.35 mmol) under ice-cooling. The reaction mixture was then stirred for another 2 h and filtered. The filter cake was washed with water and dried to give the desired product as a yellow solid (150 mg, yield: 68%).

[0527] (E)-2-(2-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)hydrazono)acetic acid (79-B) To a mixture of compound 79-A (150 mg, 0.31 mmol) in concentrated HCl (20 mL) was added 2-oxoacetic acid (80 mg, 0.54 mmol, 50 wt%). The mixture was stirred at room temperature for 2 h and filtered. The filter cake was washed with water and dried to give the desired product as a solid (130 mg, yield: 99%). ESI: [M+H] + =409.1

[0528] 4-(7-(5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (79-C) To a mixture of compound 79-B (110 mg, 0.27 mmol) in dry toluene (5 mL) was added TEA (82 mg, 0.81 mmol) and DPPA (110 mg, 0.40 mmol). The mixture was heated to reflux for 3 h. After cooling to room temperature, the reaction mixture was concentrated to remove toluene, and the residue was dissolved in DCM. The resulting solution was washed with water, brine, and dried over sodium sulfate. After filtration, the solution was concentrated and purified by silica gel chromatography (DCM:CH3OH=10:1) to give the desired compound as a solid (40 mg, 30% yield). ESI: [M+H] + =406.1

[0529] 4-(7-(4-(2-(dimethylamino)ethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (79) To a mixture of 79-C (40 mg, 0.10 mmol) in DMF (1.5 mL) was added 2-bromo-N,N-dimethylethan-1-amine hydrobromide (100 mg, 0.5 mmol) and Cs2CO3 (160 mg, 0.5 mmol). The mixture was placed in a microwave reactor at 100° C. for 2 h. The reaction mixture was concentrated and purified by preparative HPLC (TFA) to give the desired compound as a trifluoroacetate salt as a white solid (2 mg, yield: 4%). ESI: [M+H] + = 477.2; 1 H NMR (400 MHz, CD3OD): δ 8.36 (d, J = 2.4 Hz, 1H), 8.26 (dd, J = 8.9, 2.4 Hz, 1H), 8.12 (s, 1H), 8.02 (s, 1H), 7.81 (d, J = 8.9 Hz, 1H), 7.78 - 7.73 (m, 3H), 7.72 - 7.63 (m, 3H), 7.28 (s, 1H), 5.41 (s, 2H), 4.23 (t, J = 5.8 Hz, 2H), 3.60 (t, J = 5.8 Hz, 2H), 3.01 (d, J = 12.3Hz, 6H).

[0530] [ka]

[0531] tert-Butyl (R)-4-(((12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)amino)methyl)-2,2-dimethyloxazolidine-3-carboxylate (80-A) A solution of compound Int-B (800 mg, 2.37 mmol), tert-butyl (S)-4-formyl-2,2-dimethyloxazolidine-3-carboxylate (700 mg, 3.56 mmol) in methanol (40 mL) was stirred at room temperature for 1 h, then sodium cyanoborohydride (700 mg, 4.85 mmol) was added. The reaction mixture was stirred at room temperature for 1 h and quenched by the addition of aqueous NaHCO3. The mixture was extracted three times with DCM. The combined organic layers were dried over Na2SO4, filtered and concentrated to dryness. The residue was purified by silica gel chromatography (DCM:CH3OH=10:1) to give the desired compound as a white solid (300 mg, yield: 23%). ESI: [M+H] + =551.3

[0532] tert-Butyl (R)-4-((2-bromo-N-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)acetamido)methyl)-2,2-dimethyloxazolidine-3-carboxylate (80-B) To a mixture of compound 80-A (100 mg, 0.18 mmol) in DCM (10 mL), sodium hydride (48 mg, 1.06 mmol) was added under ice cooling, and the mixture was stirred at room temperature for 1 h. Before stirring at room temperature for 3 h, 2-bromoacetyl bromide (90 mg, 0.45 mmol) was added under ice cooling. The reaction mixture was poured into ice-cold 1N aqueous NaOH solution and extracted three times with DCM. The combined organic layers were dried over Na2SO4, filtered, and concentrated to give the crude product as a solid (80 mg), which was used directly in the next step. ESI: [M+H] =671.2 / 673.2

[0533] (R)-N-(2-amino-3-hydroxypropyl)-2-bromo-N-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)acetamide (80-C) To a solution of compound 80-B (80 mg, 0.12 mmol) in DCM (10 mL) was added TFA (100 mg, 0.87 mmol), and the resulting solution was stirred at room temperature for 1 h and concentrated to dryness. The residue was purified by preparative HPLC (TFA) to give the desired compound as a trifluoroacetate salt as a white solid (70 mg, yield: 90%). ESI: [M+H] + =531.1 / 533.1

[0534] (R)-4-(7-(5-(hydroxymethyl)-2-oxopiperazin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (80) A biphasic solution of compound 80-C (60 mg, 0.09 mmol) in DCM (5 mL) and aqueous NaOH (2 mL, 2N) was stirred at room temperature for 1 h. The reaction mixture was diluted with DCM and washed with water. The organic phase was dried over Na2SO4, filtered, concentrated to dryness and the residue was purified by preparative HPLC (TFA) to give the desired compound as a trifluoroacetate salt as a white solid (17 mg, yield: 34%). ESI: [M+H] + =451.2; 1 H NMR (400 MHz, CD3OD): δ 8.02 (s, 1H), 7.81 (d, J = 8.7 Hz, 1H), 7.75 (m, 4H), 7.70 (d, J = 8.6 Hz, 2H), 7.66 (s, 1H), 7.64 (dd, J = 8.6, 2.4 Hz, 1H), 7.28 (s, 1H), 5.38 (s, 2H), 4.11 (d, J = 13.1 Hz, 2H), 4.05 - 3.98 (m, 1H), 3.99 - 3.89 (m, 3H), 3.85 - 3.75 (m, 1H).

[0535] [ka]

[0536] 4-(7-Bromo-3-methyl-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile (Int-C) A solution of compound Int-A-6 (2.0 g, 5 mmol) and acetohydrazide (746 mg, 10 mmol) in dioxane (25 mL) was stirred at 110° C. for 16 h. The reaction mixture was concentrated under reduced pressure. The residue was diluted with ice water (20 mL) and extracted with hot ethyl acetate / THF (2:1, 25 mL*3). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give a dark brown solid. The solid was triturated with MeOH (10 mL) to give the desired compound (1.30 g, crude) as a pale yellow solid.

[0537] [ka]

[0538] 4-(7-(3-(2-hydroxyethyl)-2-oxopyrrolidin-1-yl)-3-methyl-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile (81-A) Compound 081-A was synthesized from compound Int-C using the procedure described above. ESI: [M+H] + = 465.2

[0539] 4-(7-(3-(2-(dimethylamino)ethyl)-2-oxopyrrolidin-1-yl)-3-methyl-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile (81) Using the procedure described above, compound 081 was synthesized. ESI: [M+H] + = 492.2; 1H NMR (400 MHz, CD3OD): δ 8.07 (dd, J = 13.2, 2.2 Hz, 1H), 7.94 - 7.85 (m, 1H), 7.72 (d, J = 8.4 Hz, 2H), 7.69 - 7.62 (m, 4H), 7.19 (d, J = 1.8 Hz, 1H), 5.23 (s, 2H), 3.98-3.95 (m, 2H), 3.50 - 3.39 (m, 1H), 3.25 (s, 1H), 2.95 (d, J = 2.2 Hz, 6H), 2.89-2.84 (m, 1H), 2.70 (s, 3H), 2.51 - 2.41 (m, 1H), 2.27-2.21 (m, 1H), 2.03 - 1.87 (m, 2H).

[0540] Using the procedures described above, the examples in Table 11 below were synthesized.

[0541] [Table 11]

[0542] [ka]

[0543] 4-(7-(3-(aminomethyl)-2-oxopyrrolidin-1-yl)-3-methyl-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile (86-A) Using the procedure described above, compound 86-A was synthesized. ESI: [M+H] + = 450.2

[0544] 4-(7-(3-((bis(methyl-d3)amino)methyl)-2-oxopyrrolidin-1-yl)-3-methyl-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile (86) To a mixture of compound 86-A (30 mg, 0.07 mmol), DCDO (30 mg, 0.94 mmol) in CD3OD (2 mL) was added NaBD3CN (30 mg, 0.94 mmol). The mixture was stirred at room temperature for 1 h. The reaction mixture was washed with water and extracted with dichloromethane. The organic phase was dried over sodium sulfate, concentrated and purified by preparative HPLC (TFA) to give the desired compound as a solid (12 mg, yield: 30%). ESI: [M+H] + = 484.3; 1 H NMR (400 MHz, CD3OD): δ 8.07 (d, J = 5.8 Hz, 1H), 7.93 (s, 1H), 7.73-7.62 (m, 6H), 7.19 (d, J = 1.7 Hz, 1H), 5.23 (s, 2H), 4.01 (d, J = 8.5 Hz, 2H), 3.64-3.58 (m, 1H), 3.38-3.31 (m, 2H), 2.69 (s, 3H), 2.57-2.46 (m, 1H), 2.04-1.94 (m, 1H)

[0545] [ka]

[0546] 4-(10-amino-7-bromo-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (Int-D-1) To a solution of compound Int-A-5 (1 g, 2.64 mmol) in DMF (20 mL) was added NaH (169 mg, 4.2 mmol, 60% purity) at 0° C. After stirring for 1 h, (aminooxy)diphenylphosphine oxide (738 mg, 3.2 mmol) was added and the mixture was stirred at 20° C. for 1 h. The mixture was poured into ice water (100 mL) and extracted with EtOAc (300 mL*3). The organic layer was washed with brine (100 mL), dried over MgSO4, and concentrated in vacuo. The residue was taken in MTBE (20 mL) and stirred for 1 h. The precipitate was collected by filtration to give the desired product (1.0 g, crude) as a yellow solid, which was used in the next step without further purification. ESI [M+H] + = 393.1 / 395.1

[0547] 4-(7-Bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazepin-12-yl)benzonitrile (Int-D) A solution of compound Int-D-1 (900 mg, 2.29 mmol) in formamide (10 mL) was stirred at 200° C. for 1.5 h. The mixture was cooled to room temperature, poured into ice water (20 mL), and filtered. The filter cake was triturated with EtOH (10 mL) for 1 h, then filtered to give the desired compound (600 mg) as a yellow solid. ESI [M+H] + = 402.1 / 404.1; 1 H NMR (400MHz, DMSO-d6): δ 8.34 (br. s., 1H), 7.93 (br. s., 1H), 7.78 (br. s., 6H), 7.40 - 7.11 (m, 2H), 5.38 (br. s., 2H).

[0548] [ka]

[0549] (S)-4-(7-(4-(hydroxymethyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazepin-12-yl)benzonitrile (87-A) Using the procedure described above, compound 87-A was synthesized as an off-white solid. ESI: [M+H] + = 437.2

[0550] (R)-4-(7-(4-((dimethylamino)methyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazepin-12-yl)benzonitrile (87) Using the procedure described above, compound 87 was synthesized. ESI: [M+H] + = 464.2; 1 H NMR (400 MHz, CD3OD): δ 8.13 (s, 1H), 8.00 (d, J = 2.4 Hz, 1H), 7.83 (d, J = 8.9 Hz, 1H), 7.67 - 7.62 (m, 3H), 7.61 - 7.56 (m, 3H), 7.17 (d, J = 1.8 Hz, 1H), 5.23 (s, 2H), 4.09 (dd, J = 9.6, 7.2 Hz, 1H), 3.69 (dd, J = 9.6, 7.2 Hz, 1H), 3.31 (d, J = 7.3 Hz, 2H), 3.05 - 2.95 (m, 1H), 2.89 (s, 6H), 2.81 (dd, J = 16.9, 8.3 Hz, 1H), 2.48 (dd, J = 16.9, 8.3 Hz, 1H).

[0551] [ka]

[0552] (S)-1-(12-(4-fluorophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-4-(hydroxymethyl)pyrrolidin-2-one (89-A) Int-E was synthesized similarly to the synthesis of Int-A. Compound 89-A was synthesized using the procedure described above. ESI: [M+H] + = 429.2

[0553] (R)-1-(12-(4-fluorophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-4-(((2-hydroxyethyl)(methyl)amino)methyl)pyrrolidin-2-one (89) Using the procedure described above, compound 89 was synthesized. ESI: [M+H] + =486.2; 1 H NMR (400 MHz, CD3OD): δ 8.21 - 7.89 (m, 3H), 7.74 (m, 2H), 7.67 - 7.46 (m, 3H), 7.20 (d, J = 1.8 Hz, 1H), 7.15 - 6.99 (m, 2H), 5.37 (s, 2H), 4.22 (dd, J = 9.7, 6.0 Hz, 1H), 3.94 (t, J = 5.3 Hz, 2H), 3.77 (d, J = 5.2 Hz, 1H), 3.48-3.46 (m, 4H), 3.29 - 3.24 (m, 1H), 3.05 (s, 3H), 2.56-2.54 (m, 1H), 2.14 - 1.93 (m, 1H).

[0554] [ka]

[0555] (R)-1-(12-(4-fluorophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-7-yl)-3-(((2-hydroxyethyl)(methyl)amino)methyl)pyrrolidin-2-one (90) Using the procedure described above, the title example was synthesized. ESI: [M+H] + =486.2; 1 H NMR (400 MHz, CD3OD): δ 8.04 (m, 2H), 7.85 (dd, J = 8.9, 2.4 Hz, 1H), 7.74 (d, J = 8.9 Hz, 1H), 7.66 (d, J = 1.6 Hz, 1H), 7.56 (td, J = 5.3, 3.0 Hz, 3H), 7.17 (d, J = 1.7 Hz, 1H), 7.12 - 6.99 (m, 2H), 5.34 (s, 2H), 4.20 (dd, J = 9.7, 8.2 Hz, 1H), 3.91 (t, J = 5.1 Hz, 2H), 3.82 (dd, J = 9.7, 7.3 Hz, 1H), 3.48 (d, J = 1.6 Hz, 1H), 3.35 (d, J = 4.2 Hz, 2H), 3.28 - 3.25 (m, 1H), 3.21 - 3.08 (m, 1H), 3.05-3.01 (m, 3H), 2.94-2.92 (m, 1H), 2.63-2.60 (m, 1H).

[0556] [ka]

[0557] 4-(7-Bromo-6-fluoro-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (91-B) Compound 91-A was synthesized similarly to the synthesis of Int-A. Compound 91-B was synthesized using the procedure described above. ESI: [M+H] + = 419.0 / 421.0

[0558] (S)-4-(6-fluoro-7-(4-(hydroxymethyl)-2-oxopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (91) Using the procedure described above, compound 91 was synthesized. ESI: [M+H] + = 481.2; 1 H NMR (400 MHz, CD3OD): δ 7.74 (d, J = 7.8 Hz, 1H), 7.72 - 7.69 (m, 3H), 7.67 - 7.63 (m, 2H), 7.55 (d, J = 11.2 Hz, 1H), 7.51 (d, J = 1.9 Hz, 1H), 7.27 (d, J = 1.5 Hz, 1H), 7.07 (d, J = 1.9 Hz, 1H), 5.18 (s, 2H), 3.97 (dd, J = 9.6, 7.7 Hz, 1H), 3.65 (dd, J = 9.6, 6.1 Hz, 1H), 2.86 (dt, J = 14.6, 7.4 Hz, 1H), 2.74 (dd, J = 17.0, 8.6 Hz, 1H), 2.55 (qd, J = 12.3, 7.5 Hz, 2H), 2.39 (dd, J = 17.0, 7.1 Hz, 1H), 2.33 (s, 6H).

[0559] [ka]

[0560] (R)-4-(7-(4-((dimethylamino)methyl)-2-oxopyrrolidin-1-yl)-8-fluoro-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (92) Using the procedure described above, compound 92 was synthesized. ESI: [M+H] + = 481.2; 1H NMR (400 MHz, CD3OD): δ 8.04 (d, J = 1.7 Hz, 1H), 7.81 (d, J = 1.7 Hz, 1H), 7.78 - 7.66 (m, 6H), 7.65 - 7.60 (m, 1H), 7.31 (d, J = 1.7 Hz, 1H), 5.49 (s, 2H), 4.08 (dd, J = 9.6, 8.0 Hz, 1H), 3.76 (dd, J = 9.5, 7.4 Hz, 1H), 3.41 (d, J = 7.6 Hz, 2H), 3.23 - 3.10 (m, 1H), 2.97 (s, 6H), 2.86 (dd, J = 17.0, 8.7 Hz, 1H), 2.54 (dd, J = 17.0, 8.4, 1H).

[0561] [ka]

[0562] Methyl 12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine-7-carboxylate (Int-AA-9) A suspension of Int-A (2.4 g, 6 mmol, 1.0 equiv), Pd(dppf)Cl2 (439 mg, 0.6 mmol, 0.05 equiv) and TEA (1.8 g, 18 mmol, 3.0 equiv) in DMF / MeOH (100 mL, 1:1) was degassed and then heated to 80-100 °C for 12 h under CO atmosphere (100 psi). The mixture was then cooled to room temperature and concentrated under reduced pressure. The resulting residue was partitioned between ethyl acetate (300 mL) and brine (300 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated to give the crude product (2.4 g), which was used in the next step without further purification.

[0563] 12-(4-Cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine-7-carboxylic acid (Int-AA) To a mixture of crude compound Int-AA-9 (1.9 g, 5 mmol, 1.0 equiv) in DMF / MeOH / THF (20 mL:10 mL:20 mL), 10% LiOH was added to adjust pH=10. The mixture was stirred at room temperature for 1 h. 1N HCl was added to adjust pH to 2-3, and the resulting precipitate was collected by filtration to give the desired acid (1.7 g) as a white solid.

[0564] [ka]

[0565] General Procedure A: 12-(4-Cyanophenyl)-N-(2-(dimethylamino)ethyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine-7-carboxamide (101) To a solution of compound Int-AA (160 mg, 0.44 mmol), DIPEA (170 mg, 1.32 mmol) and N,N-dimethylethane-1,2-diamine (39 mg, 0.48 mmol) in DCM (5 mL) was added HATU (167 mg, 0.44 mmol) and the mixture was stirred at 20° C. for 2 h under nitrogen atmosphere until TLC showed no SM remained. The reaction mixture was quenched with water (20 mL) and extracted with DCM (20 mL x 2). The combined organic layers were washed with brine (20 mL) and dried over anhydrous Na2SO4. After filtration, the solvent was concentrated under reduced pressure and the residue was purified by silica gel chromatography eluting with DCM / MeOH (100 / 1 to 20 / 1) to give the desired compound as a yellow solid (56 mg, yield: 29%). ESI: [M+H] + =437.2; 1H NMR (400 MHz, CD3OD): δ 8.21 (s, 1H), 8.08 (dd, J = 7.8, 5.6 Hz, 2H), 7.83 (d, J = 8.5 Hz, 1H), 7.78 - 7.72 (m, 3H), 7.70 (d, J = 8.3 Hz, 3H), 7.30 (s, 1H), 5.43 (s, 2H), 3.80 (t, J = 5.9 Hz, 2H), 3.41 (t, J = 5.8 Hz, 2H), 3.00 (s, 6H).

[0566] Examples 102-111 Compounds 102-111 in Table 12 were synthesized by using the procedures described above.

[0567] [Table 12] TIFF2024504927000153.tif246170

[0568] [ka]

[0569] 4-(7-(3-(hydroxymethyl)pyrrolidine-1-carbonyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (112-A) Compound 112-A was prepared by using general procedure A (45 mg, yield: 36.9%). ESI: [M+H] + = 450.2.

[0570] General Procedure B: 4-(7-(3-((dimethylamino)methyl)pyrrolidine-1-carbonyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (112) To a solution of 112-A (45 mg, 0.1 mol) and TEA (1 mL) in DCM (4 mL) was added MsCl (20 mg, 0.17 mol) at 0° C. under N2 atmosphere. The mixture was stirred for 1 h until TLC showed no SM remained. The mixture was poured into saturated aqueous NaHCO3 (20 mL) and then extracted with DCM (20 mL x 2). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, and filtered. The organic layers were concentrated under vacuum. The residue was purified by silica gel chromatography eluting with DCM / MeOH (100 / 1 to 20 / 1) to give the desired product as a solid (30 mg, yield: 57.7%). ESI [M+H] + =528.2. To a solution of the above alcohol (30 mg, 0.06 mol) in DMF (1 mL) was added dimethylamine in THF (1 mol / L, 1 mL) at 100° C. for 1 h under microwave conditions. The mixture was poured into water (20 mL) and then extracted with DCM (20 mL x 2). The combined organic layers were washed with brine (20 mL x 2), dried over Na2SO4 and filtered. The organic layers were concentrated under vacuum. The residue was purified by preparative HPLC to give the desired compound 112 as a solid (8.4 mg, yield: 31.1%). ESI: [M+H] + = 477.2; 1 H NMR (400 MHz, CD3OD): δ 8.01 - 7.99 (d, J = 10.7 Hz,1H), 7.88 - 7.86 (d, J = 7.0 Hz , 1H), 7.79 - 7.68 (m, 7H), 7.64 (m, 1H), 7.27 (s, 1H), 5.40-5.39 (d, J = 4.7 Hz ,2H), 3.81 - 3.61 (m, 4H), 3.17 (m, 2H), 2.97 (s, 3H), 2.85 (s, 3H), 2.74(m, 1H), 2.28 - 2.19 (m, 1H), 1.78-1.76 (m, 1H).

[0571] Examples 113-118 Compounds 113-118 in Table 13 were synthesized by using the same procedures described above.

[0572] [Table 13] TIFF2024504927000156.tif214170

[0573] [ka]

[0574] tert-Butyl-3-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine-7-carboxamide)azetidine-1-carboxylate (119-A) Compound 119-A was prepared by using general procedure A (30 mg, yield: 21%). ESI: [M+H] + =521.2.

[0575] General Procedure C: 2-(4-Cyanophenyl)-N-(1-methylazetidin-3-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine-7-carboxamide (119) To a stirred mixture of compound 119-A (30 mg, 0.06 mmol) in DCM (2 mL) was added TFA (1 mL) and the reaction solution was stirred at room temperature for 1 h until TLC showed no SM remained. The reaction mixture was concentrated under vacuum. The residue was dissolved in DCM (10 mL) and saturated aqueous NaHCO3 (20 mL) was added to adjust the pH to > 7. The mixture was extracted with DCM (2 x 20 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4 and filtered. The solution was concentrated under vacuum to give the amine (20 mg, crude). ESI: [M+H] + =421.2. To a solution of the above compound (20 mg, 0.05 mmol) and paraformaldehyde (15 mg, 0.5 mmol) in DCM (3 mL) and MeOH (3 mL), NaBH3CN (15 mg, 0.25 mmol) was added and the reaction mixture was stirred at room temperature for 1 h. The mixture was poured into saturated aqueous NaHCO3 (10 mL) and then extracted with DCM (2 x 10 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, and filtered. The organic solution was concentrated under vacuum and the residue was purified by preparative HPLC to give the desired compound as a white solid (10 mg, yield: 38%). ESI: [M+H] + = 435.2; 1 H NMR (400 MHz, CD3OD): δ 8.20 (s, 1H), 8.08 (s, 1H), 8.01 (s, 1H), 7.83 (d, J = 8.5 Hz, 1H), 7.72 (m, 5H), 7.63 (s, 1H), 7.26 (s, 1H), 5.42 (s, 2H), 4.84(m, 1H), 4.64 (m, 2H), 4.35 (m, 1H), 4.27 (m, 1H), 3.03 (d, J = 16.1 Hz, 3H).

[0576] [ka]

[0577] tert-Butyl (S)-2-(((methylsulfonyl)oxy)methyl)azetidine-1-carboxylate (120-A) To a solution of tert-butyl (S)-2-(hydroxymethyl)azetidine-1-carboxylate (10 g, 53.4 mmol) in THF (100 mL) was added TEA (6.47 g, 64 mmol) and TsCl (11.7 g, 61.4 mmol) at 0° C. The mixture was stirred at room temperature overnight. The mixture was filtered, and the filtrate was concentrated and purified by chromatography on silica gel eluted with PE:EA=10:1 to give the desired product (17.9 g) as a pale yellow oil.

[0578] tert-Butyl (S)-2-((bis(methyl-d3)amino)methyl)azetidine-1-carboxylate (120-B) To a solution of compound 120-A (2.53 g, 7.35 mmol) in DMA (10 mL), dimethylamine-d6 hydrochloride (3 g, 36.8 mmol) and K2CO3 (5.08 g, 36.8 mmol) were added at 25° C., and the mixture was stirred at room temperature for 30 min. The supernatant of the above mixture was transferred to a sealed tube and stirred at 75° C. overnight. After cooling to room temperature, the solution was then concentrated under reduced pressure. The residue was purified by chromatography on silica gel eluted with DCM:MeOH=20:1 to give the desired product (1.24 g, 76.8%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3): δ 4.29 (d, J = 8.0 Hz, 1H), 3.88-3.78(m, 2H), 2.76-2.72 (m, 1H), 2.57-2.51 (m, 1H), 2.34-2.33 (m, 1H), 2.06-1.98(m, 1H), 1.44 (s, 9H).

[0579] (S)-N-(azetidin-2-ylmethyl)-N-(methyl-d3)methanamine-d3(120-C) To a solution of compound 13 (1.24 g), HCl (8 M, 10 mL) was added at 0° C. The resulting solution was stirred at room temperature overnight and then washed with DCM (2×20 mL) and MTBE (2×20 mL). The aqueous layer was lyophilized to give the desired product (700 mg, 99%) in its HCl salt form as a pale yellow solid.

[0580] (S)-4-(7-(2-((bis(methyl-d3)amino)methyl)azetidine-1-carbonyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (120) A suspension of compound Int-AA (1.26 g, 3.45 mmol), compound 120-C (700 mg, 3.62 mmol), HATU (1.44 g, 3.79 mmol) and DIPEA (2.4 g, 18.63 mmol) in DMA (10 mL) was stirred at 0° C. for 10 min. After the reaction was completed, the mixture was concentrated and purified by chromatography on silica gel eluted with DCM:MeOH=20:1 to give the desired product (840 mg, 52.2%) as a white solid. ESI: [M+H] + = 469.3 1 H NMR (400 MHz, CDCl3): δ 7.83 (d, J = 1.5 Hz, 1H), 7.69 (d, J = 8.1 Hz, 1H), 7.60 (d, J = 8.5 Hz, 2H), 7.56 (d, J = 8.5 Hz, 2H), 7.45 (d, J = 8.3 Hz, 1H), 7.42 (d, J = 1.3 Hz, 1H), 7.33 (d, J = 1.3 Hz, 1H), 7.26 (s, 1H), 7.17 (d, J = 1.8 Hz, 1H), 7.11 (d, J = 1.8 Hz, 1H), 5.10 (s, 2H), 4.78 - 4.62 (m, 1H), 4.42-4.30 (m, 1H), 4.09 (dd, J = 15.2, 9.0 Hz, 1H),3.02-2.87 (m, 1H), 2.76-2.63 (m, 1H), 2.58-2.46 (m, 1H), 2.28-2.15 (s, 1H).

[0581] Examples 121-131 Compounds 121-131 in Table 14 were synthesized by using the same procedures described above.

[0582] [Table 14] TIFF2024504927000160.tif255168TIFF2024504927000161.tif79170

[0583] [ka]

[0584] tert-Butyl 3-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine-7-carboxamido)azetidine-1-carboxylate (132-A) Compound 132-A was prepared as a solid by using general procedure A (85 mg, yield: 64%). ESI: [M+H] + = 521.2

[0585] General Procedure D: tert-Butyl-3-(12-(4-cyanophenyl)-N-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine-7-carboxamide)azetidine-1-carboxylate (132-B) To a solution of compound 132-A (85 mg, 0.16 mmol) in DMF (5 mL) was added sodium hydride (15 mg, 0.64 mmol) at 0° C., and the resulting mixture was stirred for 30 min before adding MeI (40 mg, 0.28 mmol). The reaction mixture was stirred at 0° C. for 1 h and then quenched by the addition of saturated aqueous NH4Cl (20 mL). The resulting mixture was extracted with EA (2×20 mL); the combined organic layers were washed with brine (20 mL) and dried over anhydrous Na2SO4. After filtration, the organic solution was concentrated under vacuum. The residue was purified by silica gel chromatography eluting with DCM / MeOH (100 / 1 to 20 / 1) to give the desired compound (50 mg, yield: 57%). ESI [M+H] + = 535.2.

[0586] 12-(4-Cyanophenyl)-N-methyl-N-(1-methylazetidin-3-yl)-9H-benzo[e]imidazo[2,1c]pyrrolo[1,2-a][1,4]diazepine-7-carboxamide (132) Compound 132 was prepared as a solid by using general procedure C (2 mg, yield: 22%). ESI [M+H] + = 448.2; 1 H-NMR (400 MHz, CD3OD) δ 8.09 (s, 1H), 7.84 (d, J = 8.5 Hz, 2H), 7.79 - 7.66 (m, 8H), 7.33 (s, 1H), 5.44 (s, 2H), 4.71 - 4.16 (m, 4H), 3.03 (s, 6H).

[0587] Examples 133-135 Compounds 133-135 in Table 15 were synthesized by using the same procedures described above.

[0588] [Table 15]

[0589] [ka]

[0590] Compound 136 was a by-product during the synthesis of compound 135. ESI: [M+H] + = 467.2; 1 H-NMR (400 MHz, CD3OD): δ 8.43 (d, J = 1.9 Hz, 1H), 8.29 (dt, J = 10.6, 5.3 Hz, 1H), 8.07 (t, J = 4.0 Hz, 1H), 7.85 (d, J = 8.5 Hz, 1H), 7.78 - 7.65 (m, 6H), 7.31 (d, J = 1.7 Hz, 1H), 5.48 (s, 2H), 4.55 (d, J = 4.4 Hz, 2H), 3.34 (t, J = 7.6 Hz, 2H), 3.26 (d, J = 6.9 Hz, 2H), 2.81 (d, J = 10.1 Hz, 3H), 2.71-2.64 (m, 1H).

[0591] [ka]

[0592] (S)-4-(7-(2-(hydroxymethyl)azetidine-1-carbonyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (137-A) To a solution of (S)-azetidin-2-ylmethanol hydrochloride (605 mg, 4.9 mmol, 1.2 equiv.), compound Int-AA (1.5 g, 4.1 mmol, 1.0 equiv.) in dry DCM (20 mL), DIPEA (1 mL) and HATU (1.55 g, 4.1 mmol, 1.0 equiv.) were added. The mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was concentrated in vacuum. The residue was purified by flash chromatography (MeOH / DCM=0-5%) to give the desired product as a solid (1.05 g, yield: 58%). ESI: [M+H] + = 436

[0593] (S)-(1-(12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine-7-carbonyl)azetidin-2-yl)methyl methanesulfonate (137-B) To a solution of compound 137-A (1.05 g, 2.4 mmol, 1.0 equiv) in dry DCM (20 mL) was added TEA (0.67 mL, 4.8 mmol, 2.0 equiv) and MsCl (0.28 mL, 3.6 mmol, 1.5 equiv) at 0° C. The mixture was stirred at room temperature for 1 h. After the reaction was completed, the mixture was quenched with ice water (20 mL) and extracted with DCM (20 mL). The separated organic layer was washed with brine, dried over anhydrous Na2SO4, and filtered. After filtration and concentration, the crude compound was purified by flash chromatography (MeOH / DCM=0%-5%) to give the desired product as a solid (800 mg, yield: 65%). ESI: [M+H] + =513

[0594] (S)-4-(7-(2-((methylamino)methyl)azetidine-1-carbonyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (137). A mixture of compound 137-B (200 mg, 0.39 mmol, 1.0 equiv) and MeNH2 in THF (20 mL, 40 mmol) was stirred in a sealed tube at 70 °C for 16 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The resulting residue was purified by flash chromatography (MeOH / DCM = 0% to 5%), followed by preparative HPLC to give the desired product as a white solid (100 mg, yield: 56%). ESI: [M+H] + = 449; 1 H-NMR (400 MHz, DMSO-d6): δ 7.90 (d, J = 9.6 Hz, 2H), 7.78 - 7.72 (m, 6H), 7.68-7.63 (m, 2H), 7.26 (s, 1H), 7.06 (d, J = 1.8 Hz, 1H), 5.30 (s, 2H), 4.59-4.51 (m, 1H), 4.39-4.30 (m, 1H), 4.08-4.01 (m, 1H), 2.92-2.80 (m, 2H), 2.43-2.27 (m, 4H), 2.14-2.03 (m, 2H).

[0595] B. Biological Assays Materials and reagents for measuring inhibition of MK2 biochemical activity: 1) Assay plate: 384-well small volume black round bottom polystyrene NBS microplate (Corning, Cat. No. 4514) 2) MK2 Kinase Assay Kit: Z'LYTE Kinase Assay Kit (ThermoFisher, Catalog No. pv3177) 3) MAPKAPK2 recombinant human protein (ThermoFisher, Catalog No. pv3317) 4) DMSO (Vetec, catalog number V900090) 5) DTT (Sigma, catalog number D0632)

[0596] MK2 Kinase Assay Procedure: 1) Dispense 5 μl of the kinase / peptide mixture into each well of a 384-well plate. 2) Add 2.5 μl of 4X test compound to each well 3) Add 2.5 μl of 4X ATP solution to each well. 4) After 90 minutes of incubation at room temperature, add 5 μl of development solution to each well. 5) After 60 minutes of incubation at room temperature, add 5 μl of stop solution to each well. 6) The emission signals of coumarin (Ex. 400 nm, Em. 445 nm) and fluorescein (Ex. 400 nm, Em. 520 nm) are measured on an Envision plate reader (PerkinElmer).

[0597] Materials and reagents for measuring activity in THP-1 cells: 1) THP-1 cells (Shanghai Cell Bank) 2) PMA (Sigma, Catalog No. P1585) 3) LPS (Sigma, catalog number L4516) 4) 96-well tissue culture treated plates (Grenier, Cat. No. 655180) 5) FBS (Thermofisher, catalog number 10100147) 6) Complete EDTA-Free Protease Inhibitor Cocktail (Roche, Catalog No. 4693132001) 7) PhosSTOP (Roche, Catalog No. 4906845001) 8) Phosphor-Hsp27(Ser78) Sandwich Elisa Kit (CST, Cat. No. 7295)

[0598] Cell assay procedure: 1) Day 0: THP-1 cells are seeded at 50K / well in complete medium (1640 + 10% FBS + 50 μM β-ME) + 20 ng / mL PMA in 96-well plates. 2) Day 1: After 24 hours of incubation (37° C. / 5% CO2), the PMA-containing medium is removed and replaced with complete growth medium. Cells are incubated for an additional 48 hours to allow recovery. 3) Day 3: Cells are incubated with or without compounds at the indicated concentrations for 60 min before adding 20 ng / mL LPS. Cells are stimulated with LPS for 30 min, washed once with ice-cold PBS, and lysed on ice in the lysis buffer provided in the ELISA kit with protease & phosphatase inhibitors from Roche. Extracts are clarified by refrigerated centrifugation and stored at -80°C until pHsp27 / total Hsp27 ELISA. 4) Day 3 or 4: Cell Signaling Technology ELISA is performed for pSer78-Hsp27 with lysate diluted to approximately 2.5 μg / well. 5) Absorbance at 450 nm was read on a SpectraMax plus plate reader (Molecular Devices). Data was processed using GraphPad prism 8.0.

[0599] Using the procedures described above, the biological activities in Table 16 below were determined:

[0600] [Table 16] TIFF2024504927000167.tif255143TIFF2024504927000168.tif249153

[0601] Reference All publications and patents mentioned herein are incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.

[0602] Equivalent While specific embodiments of the present invention have been discussed, the above specification is illustrative and not restrictive. Numerous variations of the present invention will become apparent to those skilled in the art upon review of this specification and the following claims. The full scope of the present invention should be determined by reference to the claims and this specification, along with such variations, along with their full scope of equivalents.

Claims

1. A compound having the structure of formula (I) or a pharma- ceutically acceptable salt thereof: 【Chemistry 1】 [In the formula, X 1 , X 2 , and X 3 are each independently CH, C-(alkyl), or N; Y 1 and Y 2 are each independently H or halogen; Y 3 is CN or a halogen; Z is -C(O)NZ 1 Z 2 , -NHC(O)NZ 3 Z 4 , -NHC(O)C(HZ 5 Z 6 ), or -NZ 7 Z 8 and; Z 1 is H or alkyl; Z 2 is an optionally substituted alkyl, aminoalkyl, cycloalkyl, heterocyclyl, or heterocycloalkyl; or Z 1 and Z 2 are joined together with the N to which they are attached to form an optionally substituted 4-, 5-, or 6-membered heterocycle or an optionally substituted heterobicycle; Z 3 and Z 4 are joined together with the N to which they are attached to form an optionally substituted 4-, 5-, or 6-membered heterocycle; Z 5 and Z 6 are joined together with the C to which they are attached to form an optionally substituted 4-, 5-, or 6-membered heterocycle; Z 7 and Z 8 are linked together with the N to which they are attached to form an optionally substituted 5- or 6-membered cyclic amide, cyclic urea, or cyclic carbamate.

2. X 1 , X 2 , and X 3 The compound of claim 1, wherein at least one of is N.

3. Z is -C(O)NZ 1 Z 2 2. The compound of claim 1,

4. Z 1 is H or CH 3 and Z2 is substituted alkyl or substituted aminoalkyl; or 4. The compound of claim 3, wherein Z 1 and Z 2 , together with the N to which they are attached, join to form an optionally substituted 4-, 5-, or 6-membered heterocycle or an optionally substituted heterobicycle.

5. Z, 【Chemistry 2】 and; n is 0, 1 or 2; R 1 , R 2 , R 3 , and R 4 is independently at each occurrence H, alkyl, or hydroxyalkyl; R 5 and R 6 is, independently at each occurrence, H or alkyl.

6. Z, 【Chemistry 3】 6. The compound of claim 5,

7. Z 2 2. The compound of claim 1, wherein is optionally substituted heterocycloalkyl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl.

8. Heterocycloalkyl includes azetidinyl, pyrrolidinyl, or piperidinyl, and when substituted, is substituted with alkyl; cycloalkyl is optionally substituted cyclobutyl, cyclopentyl, or cyclohexyl, and when substituted, is substituted with alkylamino; and 8. The compound of claim 7, wherein the heterocyclyl is a substituted azetidinyl, pyrrolidinyl, or piperidinyl, and when substituted, is substituted with alkyl.

9. Z, 【Chemistry 4】 【change】 【change】 9. The compound of claim 8, wherein

10. Z 1 and Z 2 are linked together with the N to which they are attached to form an optionally substituted 4-, 5-, or 6-membered heterocycle, or an optionally substituted heterobicycle.

11. Z, 【Chemistry 5】 and; R 7 and R 8 are each independently H, CN, halo, alkyl, aminoalkyl, or alkylaminoalkyl; R 9 and R 10 are each independently H, CN, halo, hydroxyl, amino, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl; Z is 【Chemistry 6】 and; X6 is CH2, NH, or N(alkyl); R 11 and R 12 are each independently H, CN, halo, hydroxyl, amino, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl; When X 6 is NH or N(alkyl), R 11 and R 12 are not hydroxyl, amino, or alkylamino; Z, 【Chemistry 7】 and; R 13 and R 14 are each independently H, CN, halo, alkyl, aminoalkyl, or alkylaminoalkyl; R 15 and R 16 are each independently H, CN, halo, hydroxyl, amino, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl; R 15 and R 16 combine to form an optionally substituted 4-, 5-, or 6-membered nitrogen-containing heterocycle; or Z, 【Chemistry 8】 and 11. The compound of claim 10, wherein R17 is H, CN, halo, hydroxyl, amino, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl.

12. R 7 and R 8 are each independently H, alkyl, or alkylaminoalkyl; R 9 and R 10 are each independently H, alkyl, alkylamino, or alkylaminoalkyl; R 11 and R 12 are each independently H, CN, alkyl, alkylamino, or alkylaminoalkyl; R 13 and R 14 are each independently H, CN, alkyl, aminoalkyl, or alkylaminoalkyl; 12. The compound of claim 11, wherein R15 and R16 are each independently H, CN, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl, and taken together form an optionally substituted 4-, 5-, or 6-membered nitrogen-containing heterocycle.

13. Z, 【Chemistry 9】 【change】 【change】 12. The compound of claim 11, wherein

14. 【Catalog 10】 【change】 【change】 【change】 2. The compound of claim 1 having the structure:

15. Z is -NHC(O)NZ 3 Z 4 and Z 3 and Z 4 taken together with the N to which they are attached form an optionally substituted 5-membered heterocyclic ring; or The compound of claim 1, wherein Z is -NHC(O)C(HZ5Z6); and Z5 and Z6, together with the C to which they are attached, join to form an optionally substituted 5-membered cycloalkyl or heterocycle.

16. Z is 【Chemistry 11】 and; R 18 and R 19 are each independently H, CN, halo, alkyl, aminoalkyl, or alkylaminoalkyl; R 20 and R 21 are each independently H, CN, halo, hydroxyl, amino, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl; or Z is 【Chemistry 12】 and; X7 is CHR22 or NR23; R 22 is H, CN, halo, hydroxyl, amino, alkyl, aminoalkyl, alkylamino, or alkylaminoalkyl; R 23 is H, alkyl, alkylamino, or alkylaminoalkyl; preferably 16. The compound of claim 15, wherein X7 is CHR22, where R22 is alkylamino, or X7 is NR23, where R23 is H or alkyl.

17. R 20 and R 21 17. The compound of claim 16, wherein each is independently H or alkylamino, wherein alkylamino is methylamino or dimethylamino.

18. Z, 【Chemistry 13】 【change】 17. The compound of claim 16,

19. 【Catalogue 14】 3. The compound according to claim 1 or 2, having the structure:

20. Z is -NZ 7 Z 8 and Z7 and Z8, together with the N to which they are attached, combine to form an optionally substituted 5- or 6-membered cyclic amide, cyclic urea, or cyclic carbamate.

21. Z is 【Chemistry 15】 and; X 8 But, CR 28 R 29 , N.R. 30 or O; R 24 and R 26 are each independently H or alkyl; R 25 is H, or optionally substituted alkyl, aminoalkyl, alkylaminoalkyl, heterocycloalkyl, or heterocyclyl; R 27 is H, or optionally substituted alkyl, amino, alkylamino, aminoalkyl, alkylaminoalkyl, heterocycloalkyl, or heterocyclyl; R 28 and R 29 are each independently H, alkyl, alkylamino, aminoalkyl, or optionally substituted alkylaminoalkyl; R 30 is H, alkyl, alkylaminoalkyl, or optionally substituted cycloalkyl or heterocyclyl; X 8 NR 30 or O, then R 30 is not amino or alkylamino; Z is 【Chemistry 16】 and; X9 is CR36R37 or NR38; X10 is N or CR34; R 34 and R 35 are each independently H or optionally substituted alkyl; R 36 and R 37 are each independently H, alkyl, alkylamino, aminoalkyl, alkylaminoalkyl, or optionally substituted heterocyclyl; R 38 is alkyl, aminoalkyl or alkylaminoalkyl; Z, 【Chemistry 17】 and; R 39 and R 40 are each independently H or optionally substituted alkyl; R 41 is aminoalkyl, alkylaminoalkyl, or optionally substituted heterocyclyl or heterocycloalkyl; Z, 【Chemistry 18】 and; R 42 , R 43 , and R 46 are each independently H or optionally substituted alkyl; R 44 , R 45 , and R 47 are each independently H, alkylamino, aminoalkyl, or alkylaminoalkyl; Z, 【Chemistry 19】 and; R 48 , R 49 , R 51 , and R 52 are each independently H or optionally substituted alkyl; R 50 and R 53 are each independently H, alkyl, aminoalkyl, or alkylaminoalkyl; Z, 【Chemistry 20】 and; R 54 and R 55 are each independently H, alkyl, aminoalkyl, or alkylaminoalkyl; or Z, 【Chemistry 21】 and; R 54 is H, or optionally substituted alkyl or hydroxyalkyl; 21. The compound of claim 20, wherein R55 is H, alkyl, aminoalkyl, alkylaminoalkyl, or cycloalkyl.

22. R 25 and R 27 One of them is H and the other is R 25 and R 27 The other one is 【Chemical 22】 and; m is 0, 1 or 2; R 30 and R 31 are each independently H or alkyl, or R 30 and R 31 form a carbocyclic ring together with the C to which they are attached; R 32 and R 33 22. The compound of claim 21, wherein each R is independently H, alkyl, hydroxyalkyl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl, or R 32 and R 33 together with the N to which they are attached form an optionally substituted heterocycle.

23. Z, 【Chemistry 23】 【change】 【change】 22. The compound of claim 21, wherein

24. Z, 【Chemistry 24】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 22. The compound of claim 21, wherein

25.

25. 【change】 【change】 【change】 【change】 【change】 【change】 【change】 3. The compound according to claim 1 or 2, having the structure:

26. A pharmaceutical composition comprising a compound according to any one of claims 1 to 25 and a pharma- ceutically acceptable carrier.

27. Use of a compound according to any one of claims 1 to 25 or a pharmaceutical composition according to claim 26 in the manufacture of a medicament for treating or preventing a disorder associated with mitogen-activated protein kinase-activated protein kinase-2 (MK2).

28. 28. The use of claim 27, wherein the MK2 associated disorder is an inflammatory disorder or cancer, and the cancer is optionally selected from a KRAS- or BRAF-dependent cancer.

29. The use described in claim 27 or 28, wherein the MK2 associated disorder is cancer and the medicament is for co-administration or combination therapy with one or more additional chemotherapeutic agents, and the one or more additional chemotherapeutic agents are optionally selected from CHK1 inhibitors and alkylating agents.

30. Use of a compound according to any one of claims 1 to 25 or a pharmaceutical composition according to claim 26 in the manufacture of a medicament for treating or preventing a metabolic disorder.

31. 31. The use according to claim 30, wherein the metabolic disorder is diabetes, insulin resistance, obesity, or metabolic syndrome.

32. The use described in claim 30 or 31, wherein the MK2 associated disorder is diabetes or insulin resistance and the medicament is for co-administration with one or more additional antidiabetic agents, and the one or more additional antidiabetic agents include metformin.

33. A pharmaceutical composition comprising a compound according to any one of claims 1 to 25 for use in the treatment of a disorder associated with mitogen-activated protein kinase-activated protein kinase-2 (MK2).

34. The pharmaceutical composition for use according to claim 33, wherein the MK2 associated disorder is an inflammatory disorder or cancer, and the cancer is optionally selected from KRAS- or BRAF-dependent cancers.

35. A pharmaceutical composition for use according to claim 33 or 34, wherein the MK2 associated disorder is cancer and the pharmaceutical composition is for co-administration or combination therapy with one or more additional chemotherapeutic agents, the one or more additional chemotherapeutic agents optionally being selected from CHK1 inhibitors and alkylating agents.

36. A pharmaceutical composition comprising a compound according to any one of claims 1 to 25 for use in the treatment of a metabolic disorder.

37. The pharmaceutical composition for use according to claim 36, wherein the metabolic disorder is diabetes, insulin resistance, obesity, or metabolic syndrome.

38. A pharmaceutical composition for use according to claim 36 or 37, wherein the MK2 associated disorder is diabetes or insulin resistance, and the pharmaceutical composition is for co-administration with one or more additional antidiabetic agents, and the one or more additional antidiabetic agents include metformin.

Images