Fused tetracyclic compounds, compositions and uses thereof

Abstract

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

Description

[0001] Related applications

[0002] This application claims the benefit of priority to U.S. Provisional Patent Application No. 62 / 274,640, filed on January 4, 2016, which is incorporated herein by reference in its entirety. Background Art

[0003] p38 mitogen-activated protein kinase (p38MAPK) transduces a variety of extracellular signals that cause inflammatory responses, cell division and differentiation, apoptosis, and cell motility. Initially, p38MAPK was considered an ideal target for anti-inflammatory therapeutics. However, the failure of more than a dozen chemically different compounds in clinical trials suggested that p38MAPK may be a poor therapeutic target. Many of these compounds were found to have varying degrees of hepatotoxicity and to tolerate the anti-inflammatory effects that occurred within weeks. In hindsight, failure in clinical trials due to unwanted side effects may not be surprising, as p38MAPK regulates the activity of more than 60 substrates.

[0004] A downstream substrate of p38 MAPK is mitogen-activated protein kinase-activated protein kinase-2 (MAPKAPK or MK2). Among other functions, MK2 regulates the biosynthesis of tumor necrosis factor alpha and other cytokines. Furthermore, MK2 is activated following DNA damage that causes cell cycle arrest, enabling cells to repair their DNA and continue to proliferate. MK2 also phosphorylates heat shock 27 (Hsp27), a prominent biomarker for cancer progression. Therefore, MK2 may be used as a potential anti-inflammatory and anti-cancer target to improve the efficacy of chemotherapy without the unwanted side effects of affecting further upstream targets (i.e., p38 MAPK).

[0005] Therefore, there is a continuing need to discover and develop new compounds that inhibit MK2 and may be useful therapeutic agents. Summary of the Invention

[0006] In certain embodiments, the present invention relates to compounds having the structure of Formula (I) and pharmaceutically acceptable salts thereof:

[0007]

[0008] where R 1 -R 3 、X 1 -X 3 、X 5 -X 7 , Z and n are as defined in the specification.

[0009] In certain embodiments, the present invention relates to compounds having the structure of Formula (II) and pharmaceutically acceptable salts thereof:

[0010]

[0011] where R 1a 、R 1b 、R 2 、R 3 、X 1 -X 3 、X 6 、X 7 and Z are as defined in the specification.

[0012] In some embodiments, the present invention relates to pharmaceutical compositions of a compound of Formula (I) or Formula (II) and a pharmaceutically acceptable carrier.

[0013] The present invention also relates to methods of treating an MK2-associated disorder comprising administering to a subject a compound of the present invention.

[0014] 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.

[0015] The present invention also provides a method of inhibiting MK2 activity in a cell, comprising contacting the cell with a compound of the present invention.

[0016] The present invention also provides methods of treating or preventing metabolic disorders comprising administering to a subject a compound of the present invention. DETAILED DESCRIPTION

[0017] In certain aspects, the present invention provides substituted fused tetracyclic compounds and pharmaceutical compositions thereof. Specifically, the substituted fused tetracyclic compounds can be used as MK2 inhibitors, and thus can be used as anticancer agents, anti-inflammatory agents or antidiabetic agents.

[0018] I. Compounds

[0019] In certain embodiments, the present invention relates to a compound having a structure of Formula (I) or a pharmaceutically acceptable salt thereof:

[0020]

[0021] in

[0022] X 1 、X 2 and X 3 Each occurrence is independently CR 5 or N;

[0023] X 5 、X 6 and X 7Each occurrence is independently CR 7 or N;

[0024] R 1 Each occurrence is independently H, halo, -OH, -CN, or optionally substituted alkyl, alkoxy, ether, carbamate, or ester;

[0025] R 2 is H, halo, -CN, alkyl, or ester;

[0026] R 3 is H, alkyl or cycloalkyl;

[0027] R 5 is H, halo, -CN, or optionally substituted alkyl, alkoxy, aryl, heteroaryl, carbamate, or ester;

[0028] R 7 is H, halo, -OH, -CN, or optionally substituted alkyl, alkoxy, carbamate, or ester;

[0029] Z is halo or optionally substituted amino, alkylamino, heteroalkylamino, cycloalkylamino, or heterocycloalkylamino; and

[0030] n is an integer from 0 to 5.

[0031] In certain embodiments, the present invention relates to a compound having a structure of Formula (II) or a pharmaceutically acceptable salt thereof:

[0032]

[0033] where X 1 is N or CH;

[0034] X 2 and X 3 Each occurrence is independently CR 5 or N;

[0035] R 1a is H, halo, -CN, -OH or optionally substituted alkyl;

[0036] R 1b is H, halo, -CN, -OH or optionally substituted alkyl;

[0037] R 2 is H or halogen;

[0038] R 5 is H, halo, or optionally substituted alkyl; and

[0039] Z is halo or optionally substituted amino, alkylamino, heteroalkylamino, cycloalkylamino or heterocycloalkylamino.

[0040] In certain embodiments of Formulas I and II, X 1 is N; and X 2 and X 3 Is CH. In certain embodiments of Formulas I and II, X 1 and X 2 is N; and X 3 Is CH. In certain embodiments of Formulas I and II, X 1 and X 3 is N; and X 2 Is CH. In certain embodiments of Formulas I and II, X 1 、X 2 and X 3 It's N.

[0041] In certain embodiments of formula I and II, Z is halo, preferably bromo. In alternative embodiments, Z is optionally substituted amino, alkylamino, heteroalkylamino, cycloalkylamino, or heterocycloalkylamino.

[0042] In certain embodiments of Formulas I and II, Z is optionally substituted alkylamino, heterocycloalkylamino, cycloalkylamino, or -NR 8 R 9 ; and R 8 and R 9 are each independently H or optionally substituted alkyl, cycloalkyl or heterocycloalkyl; or R 8 and R 9 Together with the N to which it is attached, they combine to form an optionally substituted 4-, 5-, or 6-membered heterocyclic ring.

[0043] In certain embodiments of Formulas I and II, Z is and

[0044] R 20 、R 21 、R 22 and R 23 each independently H, halo, hydroxy, amino, or optionally substituted alkyl, alkoxy, aminoalkyl, alkylaminoalkyl, alkylamino, cycloalkyl, or heterocycloalkyl; or

[0045] R 23 and R 20 combine to form an optionally substituted 3-, 4-, 5-, or 6-membered ring;

[0046] R 21 and R 20 to form an optionally substituted 3-, 4-, 5-, or 6-membered ring; or

[0047] R 21 and R 22 In some embodiments, the optionally substituted 4-, 5-, or 6-membered ring comprises a heteroatom. In some embodiments, the heteroatom is N.

[0048] In certain embodiments of Formulas I and II, Z is

[0049]

[0050] In certain embodiments of Formulas I and II, Z is

[0051]

[0052]

[0053] In certain embodiments of Formulas I and II, Z is X 20 It's CR 24 R 26 , NH or O; and

[0054] R 24 、R 25 、R 26 and R 27 each independently H, amino, or optionally substituted alkyl, aminoalkyl, alkylaminoalkyl, alkylamino, cycloalkyl, or heterocycloalkyl; or

[0055] R 24 With R 26 In some embodiments, the optionally substituted 4-, 5-, or 6-membered ring comprises a heteroatom. In some embodiments, the heteroatom is N.

[0056] In certain embodiments of Formulas I and II, Z is

[0057]

[0058] In certain embodiments of Formulas I and II, Z is And R 28 and R 29 are each independently H, amino, or optionally substituted alkyl, alkoxy, aminoalkyl, alkylaminoalkyl, alkylamino, cycloalkyl, or heterocycloalkyl; or R 28 and R 29 In some embodiments, the optionally substituted 4-, 5-, or 6-membered ring comprises a heteroatom. In some embodiments, the heteroatom is N.

[0059] In certain embodiments of Formulas I and II, Z is

[0060]

[0061] In certain embodiments of Formulas I and II, Z is And X 21 is NH or O. In some embodiments, Z is

[0062] In certain embodiments of formulas I and II, Z is optionally substituted alkylamino, cycloalkylamino, or heterocycloalkylamino.

[0063] In certain embodiments of Formulas I and II, Z is

[0064]

[0065] In certain embodiments of Formulas I and II, Z is

[0066]

[0067] In certain embodiments of Formula I, R 1 Each occurrence is independently fluorine, chlorine, -CN, -OR 31 、-OCF3、-OC(O)-NR 31 R 32 OR-C(O)-OR 31 ; and R 31 and R 32 Each occurrence is independently optionally substituted alkyl, alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, aryl or aralkyl. In certain preferred embodiments of Formula I, R 1 is fluorine or -CN.

[0068] In certain embodiments of Formula II, R 1a and R 1b Each occurrence is independently fluorine, chlorine, -CN, -OR 31 、-OCF3、-OC(O)-NR 31 R 32 OR-C(O)-OR 31 ; and R 31 and R 32 Each occurrence is independently optionally substituted alkyl, alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, aryl or aralkyl. In certain preferred embodiments of Formula II, R 1a and R 1b Each occurrence is independently fluoro or -CN.

[0069] In certain embodiments of Formulas I and II, R 2is-C(O)-OR 41 ; and R 41 is an optionally substituted alkyl, alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, aryl, or aralkyl. In some embodiments, R 2 It is H or chlorine.

[0070] In certain embodiments of Formula I, R 3 is optionally substituted C 1-6 Alkyl or C 3-6 In some embodiments of Formula I, R 3 It’s H.

[0071] In certain embodiments of Formulas I and II, R 5 is optionally substituted alkyl, -OC(O)-NR 61 R 62 OR-C(O)-OR 61 ; R 61 and R 62 Each occurrence is independently optionally substituted alkyl, alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, aryl, or aralkyl.

[0072] In certain embodiments of Formulas I and II, R 7 Yes-OC(O)-NR 71 2 or -C(O)-OR 71 ; and R 71 is optionally substituted alkyl, alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, aryl or aralkyl.

[0073] In certain embodiments of Formula I, n is 0, 1, or 2.

[0074] In certain embodiments, the compounds of the invention may be prodrugs of compounds of Formula I or Formula II, for example, wherein a hydroxyl group in the parent compound is present as an ester or carbonate, or a carboxylic acid present in the parent compound is present as an ester. In certain such embodiments, the prodrug is metabolized in vivo to the active parent compound (e.g., an ester is hydrolyzed to the corresponding hydroxyl group or carboxylic acid).

[0075] In certain embodiments, the compounds of the present invention may be racemic. In certain embodiments, the compounds of the present invention may be enriched in one enantiomer. For example, the compounds of the present invention may have greater than 30% ee, 40% ee, 50% ee, 60% ee, 70% ee, 80% ee, 90% ee, or even 95% ee or greater. The compounds of the present invention have more than one stereocenter. Therefore, the compounds of the present invention may be enriched in one or more diastereomers. For example, the compounds of the present invention may have greater than 30% de, 40% de, 50% de, 60% de, 70% de, 80% de, 90% de, or even 95% de or greater.

[0076] In certain embodiments, as described in detail below, the present invention relates to a method for treating or preventing cancer or inflammatory conditions with a compound of Formula I or Formula II or a pharmaceutically acceptable salt thereof. In certain embodiments, therapeutic preparations can be enriched to mainly provide a kind of enantiomer of a compound (e.g., Formula I or II compound). The mixture enriched with an enantiomer can include (e.g.) at least 60mol% of a kind of enantiomer, or more preferably at least 75mol%, 90mol%, 95mol% or even 99mol%. In certain embodiments, the compound enriched with an enantiomer is substantially free of another enantiomer, wherein substantially free means that compared with the amount of another enantiomer, for example, in a composition or compound mixture, the substance accounts for less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1%. For example, if a composition or compound mixture contains 98 grams of a first enantiomer and 2 grams of a second enantiomer, it is said to contain 98 mol% of the first enantiomer and only 2% of the second enantiomer.

[0077] In certain embodiments, the therapeutic preparation can be enriched to provide primarily one diastereomer of a compound (e.g., a compound of Formula I or II). A diastereomerically enriched mixture can comprise, for example, at least 60 mol% of one diastereomer, or more preferably at least 75 mol%, 90 mol%, 95 mol%, or even 99 mol%.

[0078] In certain embodiments, the present invention provides pharmaceutical preparations suitable for treating cancer, inflammatory conditions in human patients, comprising an effective amount of any of the compounds of Formula I or Formula II and one or more pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical preparations can be used to treat or prevent conditions or diseases as described herein. In certain embodiments, the pharmaceutical preparations have sufficiently low pyrogenic activity to be suitable for use in human patients.

[0079] Compounds of any of the above structures may be used in the manufacture of a medicament for the treatment of any of the diseases or conditions disclosed herein.

[0080] Exemplary compounds of Formula I and Formula II are depicted in the Examples and Table 1. The compounds disclosed in the Examples and Table 1 should be understood to encompass both free bases and conjugate acids. For example, the compounds in the Examples and Table 1 may be depicted as complexes or salts with trifluoroacetic acid or hydrochloric acid, but compounds in their corresponding free base form or in salt form with other acids are also within the scope of the present invention. The compounds may be isolated in free base form, in salt form (e.g., hydrochloride), or in both forms. In the chemical structures shown below, standard chemical abbreviations are sometimes used.

[0081] Table 1: Exemplary compounds of Formula I and Formula II

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102]

[0103]

[0104]

[0105]

[0106]

[0107]

[0108]

[0109]

[0110]

[0111]

[0112]

[0113]

[0114]

[0115]

[0116]

[0117]

[0118]

[0119]

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127]

[0128]

[0129]

[0130]

[0131]

[0132]

[0133]

[0134]

[0135] II. Uses of MK2 Inhibitors

[0136] In certain aspects, the present invention provides methods of treating cancer comprising administering to a subject, eg, a therapeutically effective amount of a compound of Formula I or Formula II.

[0137] In certain embodiments, the cancer may be one or a variant of the following: acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), adrenocortical carcinoma, AIDS-related cancers (Kaposi Sarcoma and lymphoma), anal cancer, appendix cancer, atypical teratoid / rhabdoid tumor, basal cell carcinoma, bile duct cancer (including extrahepatic), bladder cancer, bone cancer (including osteosarcoma and malignant fibrous histiocytoma), brain tumors (e.g., astrocytomas, brain and spinal cord tumors, brain stem gliomas, central nervous system atypical teratoid / rhabdoid tumors, central nervous system embryonal tumors, craniopharyngiomas, ependymoblastomas, ependymomas, medulloblastomas, medullary epithelioma, intermediately differentiated Pineal parenchymal tumors, supratentorial primitive neuroectodermal tumors, and pineoblastomas), breast cancer, bronchogenic tumors, Burkitt lymphoma, basal cell carcinoma, bile duct cancer (including extrahepatic), bladder cancer, bone cancer (including osteosarcoma and malignant fibrous histiocytoma), carcinoid tumors, central nervous system cancers of unknown primary (such as atypical teratoid / rhabdoid tumors, embryonal tumors, and lymphomas), cervical cancer, childhood cancers, chordoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myeloproliferative disorders, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphomas (mycosis fungoides and Sézary syndrome), Syndrome), bile duct (extrahepatic), ductal carcinoma in situ (DCIS), embryonal tumor (CNS), endometrial cancer, ependymoblastoma, ependymoma, esophageal cancer, sensitive neuroblastoma, Ewing's sarcoma family of tumors, extracranial germ cell tumor, extragonadal germ cell tumor, extrahepatic bile duct cancer, eye cancer (e.g., intraocular melanoma, retinoblastoma), fibrous histiocytoma of bone (including malignant and osteosarcoma), gallbladder cancer, gastric (gastric) cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor (extracranial, extragonadal, ovarian), gestational trophoblastic tumor, glioblastoma, glioma, hairy cell leukemia, head and neck cancer, heart cancer, hepatocellular (liver) cancer, histiocytosis, Langerhans cellCell), Hodgkin's lymphoma, subpharyngeal cancer, intraocular melanoma, islet cell tumors (endocrine, pancreatic), Kaposi's sarcoma, kidney (including renal cell), Langerhans cell histiocytosis, laryngeal cancer, leukemia (including acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), hairy cell leukemia) , lip and oral cancer, liver cancer (primary), lobular carcinoma in situ (LCIS), lung cancer (non-small cell and small cell), lymphoma (AIDS-related, Burkitt's, cutaneous T-cell (mycosis fungoides and Sézary syndrome), Hodgkin's, non-Hodgkin's, primary central nervous system (CNS), male breast cancer, malignant fibrous histiocytoma and osteosarcoma of bone, medulloblastoma, medullary epithelioma, melanoma (including intraocular (eye)), Merkel cell carcinoma (Carcinoma), mesothelioma (malignant), occult primary metastatic squamous neck cancer, midline tract cancer involving the NUT gene, oral cancer, multiple endocrine neoplasia syndrome, multiple myeloma / plasma cell neoplasm, mycosis fungoides, myelodysplasia syndrome, myelodysplasia / myeloproliferative neoplasm, chronic myeloid leukemia (CML), acute myeloid leukemia (AML), myeloma and multiple myeloma, myeloproliferative disorders (chronic), nasal cavity and paranasal sinus cancer, nasopharyngeal carcinoma , neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, cancer of the mouth, oral cavity, lip and oropharynx, osteosarcoma and malignant fibrous histiocytoma of bone, ovarian cancer (e.g., epithelial, germ cell, and low malignant potential), pancreatic cancer (including islet cell tumors), papillomatosis, paraganglioma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineal parenchymal tumor of intermediate differentiation, pineoblastoma and supratentorial primitive neuroectodermal tumor, pituitary tumor, plasma cell Neoplasms / multiple myeloma, pleuropulmonary blastoma, breast cancer during pregnancy, primary central nervous system (CNS) lymphoma, prostate cancer, colorectal cancer, renal cell (kidney) cancer, renal pelvis and ureter cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcomas (e.g., Ewing's sarcoma family of tumors, Kaposi's sarcoma, soft tissue sarcoma, uterine sarcoma), Sézary syndrome, skin cancers (e.g., melanoma, Merkel cell carcinoma, non-melanoma), small cell lung cancer, small bowel cancer, soft tissue sarcoma, squamous cell carcinoma cervical cancer, occult primary squamous neck cancer, metastatic gastric cancer, supratentorial primitive neuroectodermal tumor, T-cell lymphoma (skin, mycosis fungoides, and Sézary syndrome), testicular cancer, laryngeal cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastic tumor (pregnancy), uncommon childhood cancers of unknown primary, transitional cell carcinoma of the ureter and renal pelvis, urethral cancer, uterine cancer, endometrial cancer, uterine sarcoma, Waldenstrom's macroglobulinemia ( Macroglobulinemia and Wilms Tumor.

[0138] In certain embodiments, the cancer is a KRAS-dependent or BRAF-dependent cancer.

[0139] In certain embodiments, the cancer is a solid tumor. The individual is typically one who has been diagnosed with a cancerous tumor or one who has previously been treated for a cancerous tumor (e.g., where the tumor has previously been surgically removed). The cancerous tumor can be a primary tumor and / or a secondary (e.g., metastatic) tumor.

[0140] In certain embodiments, the subject is a mammal, such as a human.

[0141] In certain embodiments, the cancer is associated with tissue of the bladder, bone marrow, breast, colon, kidney, liver, lung, ovary, pancreas, prostate, skin, or thyroid.

[0142] In certain embodiments, the method of treating cancer further comprises co-administering radiation therapy.

[0143] In some embodiments, the method of treating cancer further comprises co-administering one or more other chemotherapeutic agents.

[0144] Chemotherapeutic agents that can be administered in combination with the compounds of the present invention include: ABT-263, aminoglutethimide, amsacrine, anastrozole, asparaginase, AZD5363, Bacillus Calmette-Guérin, vaccine, BCG), bicalutamide, bleomycin, bortezomib, buserelin, busulfan, camptothecin, capecitabine, carboplatin, carfilzomib, carmustine, chlorambucil, chloroquine, cisplatin, cladribine, clodronate, cobimetinib, colchicine, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, demethoxyviridin ), dexamethasone, dichloroacetate, dienestrol, diethylstilbestrol, docetaxel, doxorubicin, epirubicin, estradiol, estramustine, etoposide, everolimus, exemestane, filgrastim, fludarabine, fludrocortisone, fluorouracil and 5-fluorouracil, fluoxymesterone, flutamide, gemcitabine, genistein, goserelin, hydroxyurea, idarubicin, ifosfamide,Imatinib, interferon, irinotecan, lenalidomide, letrozole, leucovorin, leuprolide, levamisole, lomustine, lonidamine, LY2603618, mechlorethamine, medroxyprogesterone, megestrol, melphalan, mercaptopurine, mesna, metformin, methotrexate, miltefosine, mitomycin, mitotane, mitoxantrone, MK2206, nilutamide, nocodazole, octreotide, oxaliplatin, paclitaxel, pamidronate, pentothane Statins, pazopanib, perifosine, PF-04691502, PF477736, plicamycin, pomalidomide, porfimer, procarbazine, raltitrexed, rituximab, romidepsin, selumetinib, sorafenib, streptozocin, sunitinib sunitinib, suramin, tamoxifen, temozolomide, temsirolimus, teniposide, testosterone, thalidomide, thioguanine, thiotepa, titanocenedichloride, topotecan, trametinib, trastuzumab,Retinoic acid (tretinoin), vinblastine, vincristine, vindesine, vinorelbine and vorinostat (SAHA). For example, chemotherapeutic agents that can be administered in combination with the compounds of the present invention include: amine glutethimide, amsacrine, anastrozole, asparaginase, bcg, bicalutamide, bleomycin, bortezomib, buserelin, busulfan, camptothecin, capecitabine, carboplatin, carfilzomib, carmustine, nitrogen mustard, chloroquine, cisplatin, cladribine, clodronic acid, colchicine, cyclophosphamide, cyproterone, cytarabine, dapoxet ... Carbazine, actinomycin D, daunorubicin, demethoxychloroquine, dichloroacetate, diethylstilbestrol, diethylstilbestrol, docetaxel, doxorubicin, pan-acrylic acid, estradiol, estramustine, etoposide, everolimus, exemestane, filgrastim, fludarabine, fludrocortisone, fluorouracil, flumethasone, flutamide, gemcitabine, genistein, goserelin, hydroxyurea, idarubicin, ifosfamide imatinib, interferon, irinotecan, lelidomide, letrozole, leucovorin, leuprorelin, levamisole, lomustine, lonidamine, methyldichloroethylamine, medroxyprogesterone acetate, megestrol acetate, melphalan, mercaptopurine, mesna, metformin, methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, nocodazole, octreotide, oxaliplatin, paclitaxel, pamidronate, pentothaline Statins, perifosine, plicamycin, pomadalast, porfibril sodium, procarbazine, raltitrexed, rituximab, sorafenib, streptozotocin, 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 other chemotherapeutic agent is a CHK1 inhibitor.

[0145] Many combination therapies have been developed to treat cancer. In certain embodiments, the compounds of the present invention may be administered in combination with combination therapies. Examples of combination therapies that may be administered in combination with the compounds of the present invention are included in Table 2.

[0146] Table 2: Example combination therapies for treating cancer.

[0147]

[0148]

[0149]

[0150]

[0151]

[0152]

[0153]

[0154] In some embodiments, the co-administered chemotherapeutic agent is an immuno-oncology therapeutic, such as an inhibitor of CTLA-4, indoleamine 2,3-dioxygenase, and / or PD-1 / PD-L1.

[0155] In certain embodiments, the combined administration of an MK2 inhibitor of Formula I or Formula II and one or more other therapeutic agents (e.g., one or more other chemotherapeutic agents) provides improved efficacy relative to each individual administration of the MK2 inhibitor (e.g., a compound of Formula I or II) or the one or more other therapeutic agents. In certain such embodiments, the combined administration provides an additive effect, wherein the additive effect refers to the sum of the effects of each of the individual administrations of the MK2 inhibitor and the one or more other therapeutic agents. In certain embodiments, co-administration results in a synergistic effect.

[0156] In certain embodiments, the MK2 inhibitor and one or more other chemotherapeutic agents are administered simultaneously. In alternative embodiments, the one or more other chemotherapeutic agents are administered within about 5 minutes to about 168 hours before or after administration of the MK2 inhibitor.

[0157] In certain embodiments, the present invention provides a method of inhibiting the proliferation of cancer cells, comprising contacting the cancer cells with an effective amount of a compound of Formula I or Formula II.

[0158] The present invention also provides a method for inhibiting MK2 activity in a cell, comprising contacting the cell with a compound of Formula I or Formula II. In certain embodiments, the cell is a cancer cell. The method can be performed in vivo or in vitro.

[0159] The present invention also provides methods for treating or preventing metabolic disorders, comprising administering a compound of Formula I or II to an individual. In certain embodiments, the metabolic disorder is diabetes, insulin resistance, obesity, or metabolic syndrome. In certain embodiments, the diabetes is type I, type II, or gestational diabetes. In certain embodiments, the treatment or prevention affects glycogenolysis or gluconeogenesis in an individual. In certain embodiments, the treatment or prevention reduces hepatic glucose production, hyperglycemia, fatty liver, insulin resistance, inflammation associated with insulin resistance, dyslipidemia associated with insulin resistance, or any combination thereof in an individual.

[0160] In certain embodiments, such as methods for treating diabetes, the method further comprises co-administering one or more other antidiabetic agents. Antidiabetic agents that can be co-administered with the compounds of the present invention include, but are not limited to, sulfonylureas, biguanides, α-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 specifically, antidiabetic drugs include, but are not limited to, metformin, glyburide, glimepiride, glipyride, glipizide, chlorpropamide, gliclazide, acarbose, miglitol, pioglitazone, troglitazone, rosiglitazone, isaglitazone, muraglitazole, izar), peliglitazar, sitagliptin, saxagliptin, vildagliptin, alogliptin, linagliptin, dutogliptin, dutogliptin, repaglinide, nateglinide, mitiglindine, exenatide, liraglutide, albiglutide, and insulin.

[0161] III. Pharmaceutical Compositions

[0162] In certain embodiments, the present invention provides pharmaceutical compositions comprising a compound of Formula I or II and a pharmaceutically acceptable carrier.

[0163] The compositions and methods of the present invention can be used to treat individuals in need. In certain embodiments, the individual is a mammal, such as a human or non-human mammal. When administering an animal (e.g., a human), the composition or compound is preferably administered in the form of a pharmaceutical composition comprising, for example, a compound of the present invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the industry and include, for example, aqueous solutions, such as water or physiologically buffered saline or other solvents or vehicles, such as glycols, glycerol, oils (e.g., olive oil), or injectable organic esters. In a preferred embodiment, when the pharmaceutical composition is for human administration, specifically for invasive administration routes (i.e., routes that prevent transmission or diffusion through epithelial barriers, such as injection or implantation), the aqueous solution is pyrogen-free or substantially pyrogen-free. Excipients can be selected to, for example, achieve delayed release of the agent or selectively target one or more cells, tissues, or organs. Pharmaceutical compositions can be in dosage unit form, such as tablets, capsules (including dispersed capsules and gelatin capsules), granules, lyophiles for reconstitution, powders, solutions, syrups, suppositories, injections, etc. The composition may also be presented in a transdermal delivery system, such as a skin patch. The composition may also be presented in a solution suitable for topical administration, such as eye drops.

[0164] Pharmaceutically acceptable carriers may contain physiologically acceptable agents for, for example, stabilizing a compound (e.g., a compound of the invention), increasing its solubility, or increasing its absorption. Physiologically acceptable agents include, for example, carbohydrates (e.g., glucose, sucrose, or dextran), antioxidants (e.g., ascorbic acid or glutathione), chelating agents, low molecular weight proteins, or other stabilizers or excipients. The choice of pharmaceutically acceptable carriers (including physiologically acceptable agents) depends, for example, on the route of administration of the composition. The formulation or pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (formulation) may also be a liposome or other polymer matrix, into which, for example, a compound of the invention may be incorporated. Liposomes, for example, comprising phospholipids or other lipids, are relatively simple to prepare and administer, non-toxic, physiologically acceptable, and metabolizable carriers.

[0165] As used herein, the phrase "pharmaceutically acceptable" refers to compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications, and are commensurate with a reasonable benefit / risk ratio.

[0166] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically 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 injurious to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) maltose; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, Seed oils, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, 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) ethanol; (20) phosphate buffer solution; and (21) other nontoxic compatible substances used in pharmaceutical formulations.

[0167] Pharmaceutical compositions (formulations) can be administered to a subject by any of several routes of administration, including, for example, oral (e.g., as drenches, tablets, capsules (including dispersible capsules and gelatin capsules) in the form of aqueous or non-aqueous solutions or suspensions, boluses, powders, granules, pastes for application to the tongue); absorption through the oral mucosa (e.g., sublingually); anally, rectally, or vaginally (e.g., in the form of a pessary, cream, or foam); parenterally (including intramuscularly, intravenously, subcutaneously, or intrathecally, in the form of, for example, a sterile solution or suspension); nasally; intraperitoneally; subcutaneously; transdermally (e.g., in the form of a patch applied to the skin); and topically (e.g., in the form of a cream, ointment, or spray applied to the skin or in the form of 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 appropriate routes of administration and compositions therefor can be found in, for example, U.S. Patent Nos. 6,110,973, 5,731,000, 5,541,231, 5,427,798, 5,358,970, and 4,172,896, and patents cited therein.

[0168] The formulation can be conveniently presented in unit dosage form and can be prepared by any method well known in the pharmaceutical art. The amount of active ingredient that can be combined with the carrier material to produce a single dosage form will vary depending on the subject being treated and the specific mode of administration. The amount of active ingredient that can be combined with the carrier material to produce a single dosage form should generally be the amount of compound that produces a therapeutic effect. Generally speaking, based on 100%, this amount ranges from about 1% to about 99%, preferably from about 5% to about 70%, and most preferably from about 10% to about 30% active ingredient.

[0169] The method for preparing these formulations or compositions comprises the step of mixing the active compound (e.g., a compound of the present invention) with a carrier and optionally one or more auxiliary ingredients. In general, the formulations are prepared by uniformly and intimately mixing the compound of the present invention with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, shaping the product.

[0170] The formulations of the present invention suitable for oral administration may be in the form of capsules (including dispersible capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored base, typically sucrose and gum arabic or tragacanth), lyophiles, 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 soft tablet (using an inert base, such as gelatin and glycerin, or sucrose and gum arabic) and / or as a mouthwash, etc., each containing a predetermined amount of the compound of the present invention as the active ingredient. The composition or compound may also be administered as a bolus, lick, or paste.

[0171] To prepare solid dosage forms for oral administration (capsules (including dispersible capsules and gelatin capsules), tablets, pills, sugar-coated tablets, powders, granules, etc.), the active ingredient is mixed with one or more pharmaceutically acceptable carriers (e.g., 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 carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or gum arabic; (3) humectants, such as Glycerol; (4) disintegrants such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) buffering agents 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; (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents such as modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including dispersible capsules and gelatin capsules), tablets, and pills, the pharmaceutical composition may also contain a buffering agent. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using excipients such as lactose (or milk sugar) and high molecular weight polyethylene glycols.

[0172] Tablets may be prepared by compression or molding, optionally containing one or more auxiliary ingredients. Compressed tablets may be prepared using a binder (e.g., gelatin or hydroxypropyl methylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (e.g., sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), a surfactant, or a dispersant. Molded tablets may be prepared by molding a mixture of the powdered compound moistened with an inert liquid diluent in a suitable machine.

[0173] Tablets and other solid dosage forms of pharmaceutical compositions (e.g., sugar-coated tablets, capsules (including dispersible capsules and gelatin capsules), pills, and granules) may optionally be scored or prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may also be formulated to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethylcellulose, other polymer matrices, liposomes, and / or microspheres in varying proportions to provide the desired release profile. They may be sterilized, for example, by filtration through a bacteria-retaining filter or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved in sterile water or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be compositions that release the active ingredient only or preferentially in a certain portion of the gastrointestinal tract, optionally in a delayed manner. Examples of embedding compositions that may be used include polymeric substances and waxes. If appropriate, the active ingredient may also be in microencapsulated form with one or more of the above-mentioned excipients.

[0174] Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, lyophiles for reconstitution, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, the liquid dosage form may contain inert diluents commonly used in the industry, such as water or other solvents, cyclodextrins and their derivatives; solubilizers and emulsifiers, such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (specifically, cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuranol, polyethylene glycol, and fatty acid esters of sorbitan and mixtures thereof.

[0175] 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.

[0176] Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyethylene oxide sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0177] Formulations of pharmaceutical compositions for rectal, vaginal or urethral administration may be presented as suppositories, which may be prepared by mixing one or more active compounds with one or more suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol, suppository waxes or salicylates, which are solid at room temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

[0178] The pharmaceutical composition may be formulated for administration to the oral cavity as a mouthwash, an oral spray or an oral ointment.

[0179] Alternatively or additionally, the composition can be formulated for delivery via a catheter, stent, guidewire or other intraluminal device. Delivery via such devices is particularly useful for delivery to the bladder, urethra, ureter, rectum or intestine.

[0180] Formulations 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.

[0181] Dosage forms for topical 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 pharmaceutically acceptable carrier and with any preservatives, buffers, or propellants that may be required.

[0182] Ointments, pastes, creams and gels may contain, in addition to the active compounds, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0183] Powders and sprays may 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. Sprays may additionally contain customary propellants, such as chlorofluorohydrocarbons and unsubstituted volatile hydrocarbons, such as butane and propane.

[0184] Transdermal patches have the additional advantage of providing controlled delivery of the compounds of the invention to the body. Such dosage forms can be prepared by dissolving or dispersing the active compound in an appropriate medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by providing a rate-controlling membrane or dispersing the compound in a polymer matrix or gel.

[0185] Ophthalmic formulations, ophthalmic ointments, powders, solutions, and the like are also encompassed within the scope of the present invention. Exemplary ophthalmic formulations are described in U.S. Publication Nos. 2005 / 0080056, 2005 / 0059744, 2005 / 0031697, and 2005 / 004074, and U.S. Patent No. 6,583,124, the contents of which are incorporated herein by reference. If desired, the liquid ophthalmic formulation has properties similar to those of tears, aqueous humor, or vitreous humor, or is compatible with such fluids. The preferred route of administration is topical administration (e.g., topical administration, such as eye drops, or administration via an implant).

[0186] As used herein, the phrases "parenteral administration" and "administered parenterally" refer to modes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnical, intraspinal, and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration comprise one or more active compounds in combination with one or more pharmaceutically acceptable sterile, isotonic aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions; or sterile powders that can be reconstituted into sterile injectable solutions or dispersions immediately before use, which may contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0187] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils (e.g., olive oil), and injectable organic esters (e.g., ethyl oleate). Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by maintaining the required particle size (in the case of dispersions), and by the use of surfactants.

[0188] These compositions may also contain adjuvants, such as preservatives, wetting agents, emulsifiers, and dispersants. Prevention of microbial action can be ensured by including various antibacterial and antifungal agents, such as p-hydroxybenzoic acid, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like, in the compositions. In addition, prolonged absorption of injectable pharmaceutical forms can be achieved by including agents that delay absorption, such as aluminum monostearate and gelatin.

[0189] In some cases, to prolong the effect of a drug, it is necessary to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous materials with poor water solubility. The rate of drug absorption depends on its dissolution rate, which in turn can depend on the crystal size and crystalline form. Alternatively, delayed absorption of the drug can be achieved by dissolving or suspending the drug form administered parenterally in an oil vehicle.

[0190] Injectable depot forms are prepared by forming a microencapsulation matrix of the target compound in a biodegradable polymer (e.g., polylactide-polyglycolide). Depending on the ratio of drug to polymer and the properties of the specific polymer used, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations can also be prepared by encapsulating the drug in liposomes or microemulsions that are compatible with body tissues.

[0191] For use in the methods of the invention, the active compound can be administered per se or in the form of a pharmaceutical composition containing, for example, 0.1% to 99.5% (more preferably 0.5% to 90%) active ingredient in combination with a pharmaceutically acceptable carrier.

[0192] The introduction method can also be provided by a replaceable or biodegradable device. In recent years, various slow-release polymeric devices have been developed and tested in vivo for the controlled delivery of drugs (including protein biopharmaceuticals). Various biocompatible polymers (including hydrogels) (including biodegradable and non-degradable polymers) can be used to form implants for sustained release of compounds at specific target sites.

[0193] Actual dosage amounts of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0194] The selected dosage value will depend upon a wide variety of factors including the activity of the particular compound or combination of compounds employed, or their esters, salts or amides, the route of administration, the time of administration, the rate of excretion of the particular compound employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0195] A physician or veterinarian with ordinary skill in the art can readily determine and prescribe the therapeutically effective amount of the desired pharmaceutical composition. For example, to achieve the desired therapeutic effect, the physician or veterinarian may initially start the dosage of the pharmaceutical composition or compound at a lower value than required and gradually increase the dosage until the desired effect is achieved. A "therapeutically effective amount" means a concentration of the compound sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of a compound will vary depending on the individual's weight, sex, and medical history. Other factors affecting the effective amount may include, but are not limited to, the severity of the patient's condition, the condition being treated, stabilizers for the compound, and, if necessary, another type of therapeutic agent administered with the compound of the invention. A larger total dose may be delivered by multiple administrations 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, 13th ed., 1814-1882, incorporated herein by reference).

[0196] In general, a suitable daily dose of the active compound used in the compositions and methods of the invention will be that amount of the compound which is the lowest dose effective to produce a therapeutic effect. The effective dose will generally be determined based on the factors described above.

[0197] If desired, the effective daily dose of the active compound may be administered in 1, 2, 3, 4, 5, 6 or more sub-doses administered at appropriate intervals throughout the day, optionally divided into unit dosage forms. In certain embodiments of the invention, the active compound may be administered twice or three times daily. In a preferred embodiment, the active compound will be administered once daily.

[0198] Generally, the patient receiving such treatment is any animal in need thereof, including primates (particularly humans) and other mammals (eg, horses, cattle, pigs, and sheep); and poultry and pets.

[0199] In certain embodiments, the compounds of the present invention can be used alone or in combination with another type of therapeutic agent. As used herein, the phrase "combination administration" refers to any form of administration of two or more different therapeutic compounds, such that a second compound is administered while the previously administered therapeutic compound is still effective in the body (e.g., two compounds are simultaneously effective in the patient, which may include a synergistic effect of the two compounds). For example, different therapeutic compounds may be administered in parallel or sequentially in the same formulation or in 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, individuals receiving such treatment may benefit from the combined effects of different therapeutic compounds.

[0200] In certain embodiments, the combined administration of a compound of the invention and one or more other therapeutic agents (e.g., one or more other chemotherapeutic agents) provides improved efficacy relative to each individual administration of a compound of the invention (e.g., a compound of Formula I or Ia) or one or more other therapeutic agents. In certain such embodiments, the combined administration provides an additive effect, wherein the additive effect refers to the sum of the effects of each of the individual administrations of the compound of the invention and the one or more other therapeutic agents.

[0201] The present invention includes the use of the pharmaceutically acceptable salts of the compounds of the present invention in the compositions and methods of the present invention. As used herein, the term "pharmaceutically acceptable salt" includes salts derived from inorganic or organic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, phosphoric acid, formic acid, acetic 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 molecule comprising the salt is not 1:1. For example, a salt may include one or more inorganic or organic acid molecules / base molecules, such as two hydrochloric acid molecules / Formula I or Formula II compound molecules. As another example, a salt may include less than one inorganic or organic acid molecule / base molecule, such as two Formula I or Formula II compound molecules / tartaric acid molecules.

[0202] In other embodiments, the contemplated salts of the present invention include, but are not limited to, alkyl, dialkyl, trialkyl, or tetraalkylammonium salts. In certain embodiments, the contemplated salts of the present invention include, but are not limited to, L-arginine, benzylamine, 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, the contemplated salts of the present invention include, but are not limited to, Na, Ca, K, Mg, Zn, or other metal salts.

[0203] Pharmaceutically acceptable acid addition salts may also exist in the form of various solvates, for example with water, methanol, ethanol, dimethylformamide, etc. Mixtures of such solvates may also be prepared. The solvate may originate from the solvent of crystallization, be inherent in the solvent of preparation or crystallization, or be exogenous to the solvent.

[0204] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0205] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and (3) metal chelators, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0206] IV. Definitions

[0207] The term "acyl" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0208] The term "acylamino" is art-recognized and refers to an amino group substituted with an acyl group and can be represented, for example, by the formula hydrocarbyl C(O)NH-.

[0209] The term "acyloxy" is art-recognized and refers to a group represented by the general formula hydrocarbyl C(O)O-, preferably alkyl C(O)O-.

[0210] 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.

[0211] The term "cycloalkyloxy" refers to a cycloalkyl group having an oxygen attached thereto.

[0212] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and can be represented by the general formula alkyl-O-alkyl.

[0213] The term "alkylaminoalkyl" refers to an alkyl group substituted with an alkylamino group.

[0214] As used herein, the term "alkenyl" refers to an aliphatic group containing at least one double bond and is intended to include both "unsubstituted alkenyls" and "substituted alkenyls," the latter of which refers to alkenyl moieties having substituents replacing hydrogen on one or more carbons of the alkenyl group. The substituents may occur on one or more carbons that are included or excluded from the one or more double bonds. In addition, the substituents include all those contemplated for alkyl groups, as discussed below, except where prohibited by stability. For example, substitution of an alkenyl group by one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

[0215] "Alkyl" or "alkane" is a fully saturated, straight-chain or branched, non-aromatic hydrocarbon. Generally, unless otherwise defined, a straight-chain or branched alkyl group has 1 to about 20, preferably 1 to about 10, carbon atoms. Examples of straight-chain and branched alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, amyl, and octyl. C1-C6 straight-chain or branched alkyl groups are also referred to as "lower alkyl groups."

[0216] In addition, 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 alkyl moieties having substituents replacing hydrogen on one or more carbons in the hydrocarbon backbone. If not otherwise specified, such substituents may include, for example, halogen, hydroxy, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., 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. Those skilled in the art will understand that substituted moieties on the hydrocarbon chain may themselves be substituted as desired. For example, substituents of substituted alkyl groups may include substituted and unsubstituted amino, azido, imino, amido, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamido, sulfamoyl, and sulfonates), and silicon groups, as well as ethers, alkylthiols, carbonyls (including ketones, aldehydes, carboxylates, and esters), -CF 3 , -CN, and the like. Exemplary substituted alkyl groups are described below. Cycloalkyl groups may be further substituted with alkyl, alkenyl, alkoxy, alkylthio, aminoalkyl, carbonyl-substituted alkyl, -CF 3 , -CN, and the like.

[0217] The term "C x-y " when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is intended to include groups containing x to y carbons in the chain. For example, the term "C x-y "Alkyl" refers to a substituted or unsubstituted saturated hydrocarbon group (including straight-chain alkyl and branched-chain alkyl) containing x to y carbons in the chain, including halogenated alkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl. C0 alkyl indicates hydrogen (if the group is at the terminal position) or a bond (if it is internal). The term "C 2-y Alkenyl" and "C 2-y "Alkynyl" refers to substituted or unsubstituted unsaturated aliphatic analogs analogous in length and possible substitution to the alkyl groups described above, but containing at least one double or triple bond, respectively.

[0218] As used herein, the term "alkylamino" refers to an amino group substituted with at least one alkyl group.

[0219] As used herein, the term "alkylthio" refers to a thiol group substituted with an alkyl group and can be represented by the general formula alkylS-.

[0220] As used herein, the term "alkynyl" refers to an aliphatic group containing at least one triple bond and is intended to include both "unsubstituted alkynyls" and "substituted alkynyls," the latter of which refers to alkynyl moieties having substituents replacing hydrogen on one or more carbons of the alkynyl group. The substituents may occur on one or more carbons that are included or excluded from the one or more triple bonds. Furthermore, the substituents include all those contemplated for alkyl groups, as discussed above, except where prohibited by stability. For example, substitution of an alkynyl group by one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

[0221] As used herein, the term "amide" refers to a group

[0222]

[0223] Each of the R 10 independently represent hydrogen or a hydrocarbon group, or two R 10 Together with the nitrogen atom to which it is attached, it forms a heterocyclic ring having 4 to 8 atoms in the ring structure.

[0224] The terms "amine" and "amino" are art-recognized and refer to unsubstituted and substituted amines and salts thereof, such as the moiety represented by:

[0225]

[0226] Each of the R 10 independently represent hydrogen or a hydrocarbon group, or two R 10 Together with the nitrogen atom to which it is attached, it forms a heterocyclic ring having 4 to 8 atoms in the ring structure.

[0227] As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group.

[0228] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group.

[0229] As used herein, the term "aryl" includes substituted or unsubstituted monocyclic aromatic groups in which every atom in 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 rings, wherein two or more carbon atoms are shared by two adjacent rings, wherein at least one of the rings is aromatic, for example, the other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

[0230] The term "carbamate" is industry recognized and refers to the group

[0231]

[0232] where R 9 and R 10 independently represents hydrogen or a hydrocarbon group (such as an alkyl group), or R 9 and R 10 Together with the intervening atoms, it forms a heterocycle having 4 to 8 carbons in the ring structure.

[0233] As used herein, the terms "carbocycle" and "carbocyclic" refer to a saturated or unsaturated ring in which every atom in the ring is carbon. The term carbocycle includes both aromatic carbocycles and non-aromatic carbocycles. Non-aromatic carbocycles include cycloalkane rings in which all carbon atoms are saturated and cycloolefins containing at least one double bond. "Carbocycle" includes 5- to 7-membered monocyclic rings and 8- to 12-membered bicyclic rings. Each ring of a bicyclic carbocycle can be selected from saturated, unsaturated, and aromatic rings. Carbocycles include bicyclic molecules in which two rings share 1, 2, or 3 or more atoms. The term "fused carbocycle" refers to a bicyclic carbocycle in which each ring shares two adjacent atoms with another ring. Each ring in a fused carbocycle can be selected from saturated, unsaturated, and aromatic rings. In exemplary embodiments, an aromatic ring (e.g., phenyl) can be fused to a saturated or unsaturated ring, such as cyclohexane, cyclopentane, or cyclohexene. The definition of carbocycle includes any combination of saturated, unsaturated, and aromatic bicyclic rings, if 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 decahydronaphthalene, 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 substituted at any one or more positions capable of bearing a hydrogen atom.

[0234] "Cycloalkyl" is a fully saturated cyclic hydrocarbon. "Cycloalkyl" includes monocyclic and bicyclic rings. Typically, unless otherwise defined, monocyclic cycloalkyls have 3 to about 10 carbon atoms, more typically 3 to 8 carbon atoms. The second ring of a bicyclic cycloalkyl can be selected from saturated, unsaturated, and aromatic rings. Cycloalkyls include bicyclic molecules in which the two rings share 1, 2, or 3 or more atoms. The term "fused cycloalkyl" refers to a bicyclic cycloalkyl in which each ring shares two adjacent atoms with another ring. The second ring of a fused bicyclic cycloalkyl can be selected from saturated, unsaturated, and aromatic rings. "Cycloalkenyl" is a cyclic hydrocarbon containing one or more double bonds.

[0235] The term "carbocyclylalkyl" as used herein refers to an alkyl group substituted with a carbocyclyl group.

[0236] The term "carbonate" is industry recognized and refers to the group -OCO2-R 10 , where R 10 Represents a hydrocarbon group.

[0237] As used herein, the term "carboxyl" refers to a group represented by the formula -CO2H.

[0238] As used herein, the term "ester" refers to the group -C(O)OR 10 , where R 10 Represents a hydrocarbon group.

[0239] As used herein, the term "ether" refers to a hydrocarbon group attached to another hydrocarbon group through an oxygen. Thus, an ether substituent of a hydrocarbon group may be hydrocarbon-O-. Ethers may be symmetrical or asymmetrical. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include "alkoxyalkyl" groups, which may be represented by the general formula alkyl-O-alkyl.

[0240]

[0046] The terms "halo" and "halogen" as used herein mean halogen and include chlorine, fluorine, bromine, and iodine.

[0241] As used herein, the terms "hetaralkyl" and "heteroaralkyl" refer to an alkyl group substituted with a heteroaryl group.

[0242] As used herein, the term "heteroalkyl" refers to a saturated or unsaturated chain of carbon atoms and at least one heteroatom, wherein the two heteroatoms are not adjacent.

[0243] The term "heteroalkylamino" as used herein refers to an amino group substituted with a heteroalkyl group.

[0244] The terms "heteroaryl" and "heteroaryl" include substituted or unsubstituted aromatic monocyclic ring structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, whose ring structures include at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more rings, wherein two or more carbon atoms are shared by two adjacent rings, and at least one of the rings is a heteroaromatic ring, for example, the other rings can be 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.

[0245] As used herein, the term "heteroatom" means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen and sulfur.

[0246] The terms "heterocyclyl," "heterocycle," and "heterocyclic" refer to substituted or unsubstituted non-aromatic ring structures, preferably 3-10 membered rings, more preferably 3-7 membered rings, whose ring structures include at least one heteroatom, preferably 1-4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heterocyclyl" and "heterocycle" also include polycyclic ring systems having two or more rings, wherein two or more carbon atoms are shared by two adjacent rings, wherein at least one of the rings is a heterocycle, for example, the other rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactone, lactam, and the like. Heterocyclyl groups may also be substituted with oxo groups. For example, "heterocyclyl" encompasses both pyrrolidine and pyrrolidone.

[0247] The term "heterocycloalkyl" as used herein refers to an alkyl group substituted with a heterocyclo group.

[0248] The term "heterocycloalkylamino" as used herein refers to an amino group substituted with a heterocycloalkyl group.

[0249] As used herein, the term "hydrocarbyl" refers to a group that is bonded through a carbon atom and does not have a =O or =S substituent, and which typically has at least one carbon-hydrogen bond and a (primarily) carbon backbone, but may optionally include heteroatoms. Thus, for the purposes of this application, groups such as methyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered hydrocarbyl groups, but substituents such as acetyl (which has a =O substituent on the attached carbon) and ethoxy (which is attached through an oxygen rather than a carbon) are not hydrocarbyl groups. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocyclic, heterocyclic, alkyl, alkenyl, alkynyl, and combinations thereof.

[0250] The term "hydroxyalkyl" as used herein refers to an alkyl group substituted with a hydroxy group.

[0251] The term "lower" when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is intended to include groups in which there are 10 or fewer, preferably 6 or fewer, non-hydrogen atoms in the substituent. "Lower alkyl" refers to, for example, an alkyl group containing 10 or fewer, preferably 6 or fewer carbon atoms. In certain embodiments, an acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituent as defined herein is a lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy group, respectively, whether alone or in combination with other substituents, such as those listed in the hydroxyalkyl and aralkyl groups (in such cases, atoms in the aryl group are not counted when counting the carbon atoms in the alkyl substituent).

[0252] As used herein, the term "oxo" refers to a carbonyl group. When an oxo substituent occurs 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 referred to as being substituted with "oxo," this can mean that a carbonyl moiety (i.e., -C(=O)-) is replaced by a methylene unit (i.e., -CH2-).

[0253] 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 shared by two adjacent rings, e.g., the rings are "fused rings." Each ring of the polycyclic ring may be substituted or unsubstituted. In certain embodiments, each ring of the polycyclic ring contains 3 to 10, preferably 5 to 7, atoms in the ring.

[0254] The term "silyl" refers to a silicon moiety to which are attached three hydrocarbyl moieties.

[0255] The term "substituted" refers to a moiety having a substituent that replaces a hydrogen on one or more carbons in the main chain. It should be understood that "substituted" or "substituted by..." includes an implicit premise, that is, the substitution should be consistent with the allowed valence of the substituted atom and the substituent, and the substitution will produce a stable compound, for example, it will not spontaneously transform (for example, by rearrangement, cyclization, elimination, etc.). As used herein, the term "substituted" is intended to include all allowed substituents of organic compounds. In a broad sense, the allowed substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. For appropriate organic compounds, the allowed substituents may be one or more and may be the same or different. For the purposes of the present invention, a heteroatom (such as nitrogen) may have a hydrogen substituent that meets the heteroatom valence requirements and / or any allowed organic compound substituent described herein. Substituents may include any substituent described herein, such as halogen, hydroxy, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., 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 substituents may themselves be substituted as desired. Unless specifically stated as "unsubstituted," chemical moieties referred to herein should be understood to include substituted variations. For example, a reference to an "aryl" group or moiety implicitly includes both substituted and unsubstituted variations.

[0256] The term "sulfate" is art-recognized and refers to the group -OSO3H or a pharmaceutically acceptable salt thereof.

[0257] The term "sulfonamide" is art-recognized and refers to a group represented by the following general formula:

[0258]

[0259] where R 9 and R 10 independently represents hydrogen or a hydrocarbon group (such as an alkyl group), or R 9 and R 10 Together with the intervening atoms, it forms a heterocycle having 4 to 8 carbons in the ring structure.

[0260] The term "sulfoxide" is art-recognized and refers to the group -S(O)-R 10 , where R 10 Represents a hydrocarbon group.

[0261] The term "sulfonate" is art-recognized and refers to the group SO3H or a pharmaceutically acceptable salt thereof.

[0262] The term "sulfone" is industry recognized and refers to the group -S(O)2-R 10 , where R 10 Represents a hydrocarbon group.

[0263] The term "thioalkyl" as used herein refers to an alkyl group substituted with a thiol group.

[0264] As used herein, the term "thioester" refers to the group -C(O)SR 10 or -SC(O)R 10 , where R 10 Represents a hydrocarbon group.

[0265] The term "thioether" as used herein is equivalent to an ether in which the oxygen is replaced by sulfur.

[0266] The term "urea" is recognized in the industry and can be represented by the general formula

[0267]

[0268] where R 9 and R 10 independently represents hydrogen or a hydrocarbon group (such as an alkyl group), or R 9 and R 10 Each occurrence, taken together with the intervening atoms, forms a heterocycle having from 4 to 8 carbons in the ring structure.

[0269] " blocking group " refers to the group of atoms that shields, reduces or prevents the reactivity of functional groups when being attached to the reactive functional groups in the molecule. Generally, if necessary, blocking groups can be selectively removed during the building-up process. Examples of blocking groups can be found in Greene and Wuts, Protective Groups in Organic Chemistry, 3rd edition, 1999, John Wiley & Sons, New York (NY) and Harrison et al., Compendium of Synthetic Organic Methods, Vol. 1-8, 1971-1996, John Wiley & Sons, New York. 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"), nitro-veratryloxycarbonyl ("NVOC"), and the like. Representative hydroxy protecting groups include, but are not limited to, those in which the hydroxy 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 (e.g., ethylene glycol and propylene glycol derivatives), and allyl ethers.

[0270] As used herein, a therapeutic agent that "prevents" a disorder or condition refers to a compound that, in a statistical sample, reduces the incidence of the disorder or condition in treated samples relative to untreated control samples, or delays the onset of or reduces the severity of one or more symptoms of the disorder or condition relative to untreated control samples.

[0271] The term "treatment" includes prophylactic and / or therapeutic treatment. The terms "prophylactic or therapeutic" treatment are recognized in the art and include administering one or more target compositions to a subject. If it is administered before the clinical expression of an unwanted condition (e.g., a disease or other unwanted state of the host animal), then the treatment is prophylactic (i.e., it protects the host from developing the unwanted condition), while if it is administered after the expression of the unwanted condition, then the treatment is therapeutic (i.e., it is intended to reduce, ameliorate, or stabilize the existing unwanted condition or its side effects).

[0272] The term "prodrug" is intended to encompass compounds (e.g., compounds of formula I) that are converted under physiological conditions into therapeutically active agents of the present invention. A common method for preparing a prodrug 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 enzymatic 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 represented above may be replaced by corresponding suitable prodrugs, for example, wherein the hydroxyl groups in the parent compound are presented as esters or carbonates, or the carboxylic acids present in the parent compound are presented as esters.

[0273] The term "MK2-associated disorder" is a disorder or condition in which MK2 plays a role in the incidence or symptoms of the disease or disorder. For example, MK2-associated disorders include, but are not limited to, inflammatory diseases, autoimmune diseases, destructive bone disorders, proliferative disorders, angiogenic disorders, infectious diseases, neurodegenerative diseases, and viral diseases.

[0274] 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 incidence 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 prognosis or survival time of a patient with the disease or disorder.

[0275] Examples

[0276] Examples of compounds of formula (I) or formula (II) or pharmaceutically acceptable salts thereof having useful biological activity are described below. The preparation of these compounds can be achieved by those skilled in the art of organic synthesis using known techniques and methods.

[0277] A. Chemical synthesis

[0278] The following describes the general procedures used in the methods for preparing the compounds of the present invention.

[0279] Solution 1

[0280]

[0281] 1-tert-Butyl 2-methyl 4-bromo-1H-pyrrole-1,2-dicarboxylate, S2. To a solution of methyl 4-bromo-1H-pyrrole-2-carboxylate (120 g, 588 mmol, 1.0 eq.) in MeCN (1200 mL) at 0°C was added DMAP (7.2 g, 58 mmol, 0.1 eq.), followed by (Boc)2O (141 g, 647 mmol, 1.1 eq.). The mixture was stirred at 25°C for 2 hours. The mixture was concentrated and the residue was dissolved in ethyl acetate (2.0 L). The solution was washed with glacial 1M HCl (200 mL*2), saturated aqueous NaHCO3 (200 mL), and brine (100 mL). The organic phase was dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give crude 1-tert-butyl 2-methyl 4-bromo-1H-pyrrole-1,2-dicarboxylate as a yellow oil (174 g), which was used in the next step without further purification. TLC: R f =0.77 (petroleum ether:ethyl acetate=5:1). 1H NMR (400 MHz, chloroform-d) δ=7.29 (d, J=1.8 Hz, 1H), 6.77 (d, J=1.3 Hz, 1H), 3.82 (s, 3H), 1.56 (s, 9H).

[0282]

[0283] 4-(4-Cyanophenyl)-1H-pyrrole-1,2-dicarboxylic acid 1-tert-butyl 2-methyl ester, S3. A suspension of 4-bromo-1H-pyrrole-1,2-dicarboxylic acid 1-tert-butyl 2-methyl ester (131 g, 432 mmol, 1.0 eq.), (4-cyanophenyl)boronic acid (95.3 g, 648.5 mmol, 1.5 eq.), Na2CO3 (91 g, 864 mmol, 2.0 eq) and (Boc)2O (141 g, 648 mmol, 1.5 eq.), Pd(dppf)Cl2 (15.8 g, 21.6 mmol, 0.05 eq.) in dioxane / H2O (4.9 L, 10:1) was degassed and then heated to 80°C to 100°C under N2 for 12 hr. The mixture was concentrated under reduced pressure and the residue was partitioned between ethyl acetate (3.0 L) and brine (500 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to yield crude 1-tert-butyl 2-methyl 4-(4-cyanophenyl)-1H-pyrrole-1,2-dicarboxylate as a yellow oil (150 g), which was used in the next step without further purification. ESI [M+H] = 327.2

[0284]

[0285] Methyl 4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate, S4. A solution of 1-tert-butyl 2-methyl 4-(4-cyanophenyl)-1H-pyrrole-1,2-dicarboxylate (150 g, crude) in TFA (500 mL) was stirred at 50 ° C for 2 hours. The solution was concentrated under reduced pressure. MeOH (300 mL) was added to the residue and stirred for 1 hour. The precipitate was collected by filtration to give methyl 4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (80 g, 353 mmol) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ = 12.30 (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). ESI[M+H]=227.1

[0286] Option 2

[0287]

[0288] 5-Bromo-2-nitrobenzoic acid, S6. A suspension of 3-bromobenzoic acid (249 g, 1.24 mol, 1.0 eq.) in concentrated sulfuric acid (800 mL) was cooled at 0°C. Concentrated nitric acid (57.1 mL, 1.2 mol) was added dropwise and the mixture was warmed to 20°C and stirred for 1 hr. The mixture was poured into ice water (2 L). The precipitate was collected by filtration, washed with water (1000 mL*2), and dried to yield 5-bromo-2-nitrobenzoic acid (280 g, 1.14 mol, 91.94% yield) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ = 14.12 (br. s., 1H), 8.00 (d, J = 1.8 Hz, 1H), 7.98-7.90 (m, 2H).

[0289]

[0290] (5-Bromo-2-nitrophenyl)methanol, S7. To a solution of 5-bromo-2-nitrobenzoic acid (280 g, 1.14 mol, 1.0 eq.) in THF (1.5 L) was added dropwise a 10 M solution of BH3-Me2S in THF (119.7 mL, 1.05 eq.) over 30 minutes. After addition, the mixture was heated to 70 ° C and maintained for 3 hours. The mixture was cooled to 0 ° C, slowly quenched by MeOH (100 mL) and then concentrated in vacuo. Petroleum ether / ethyl acetate (1000 mL, 10: 1) was added to the residue and stirred for 1 hour. The precipitate was collected by filtration to produce (5-bromo-2-nitrophenyl)methanol (160 g, 689 mmol, 60.49% yield) as a white solid. 1H NMR (400 MHz, chloroform-d) 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). ESI [M+H] = 232.2 / 234.2

[0291]

[0292] Methyl 1-(5-bromo-2-nitrobenzyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate, S8. To a solution of (5-bromo-2-nitrophenyl)methanol (80 g, 344 mmol, 1.0 eq.) and TEA (48.8 g, 482 mmol, 1.40 eq.) in DCM (1.5 L) was added MsCl (41.4 g, 362 mmol, 1.05 eq.) at 0 °C and the mixture was stirred at 20 °C for 0.5 hr. To the solution was then added methyl 4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (74.1 g, 327 mmol, 0.95 eq.) at 0°C, followed by a solution of tetrabutylammonium hydroxide (35.8 g, 34.5 mmol, 0.1 eq.) and NaOH (69 g, 1724 mmol, 5.0 eq.) in H2O (210 mL). The mixture was stirred at 20°C for 10 hr and then diluted with ice water (500 mL) and DCM (2000 mL). The organic layer was separated, dried over anhydrous Na2SO4, filtered, and concentrated. EtOH (200 mL) was added to the residue and stirred for 1 hr. The precipitate was collected by filtration to produce methyl 1-(5-bromo-2-nitrobenzyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (127 g, 239 mmol, 69.3% yield) as an off-white solid. TLC:R f=0.32 (petroleum ether:ethyl acetate=4:1). 1H NMR (400 MHz, 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). ESI [M+H]=440.1 / 442.1

[0293]

[0294] Methyl 1-(2-amino-5-bromobenzyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate, S9. A suspension of methyl 1-(5-bromo-2-nitrobenzyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (125 g, 284 mmol, 1.00 eq.), Fe (79 g, 1420 mmol, 5.0 eq.) and NHCl (76 g, 1420 mmol, 5.0 eq.) in EtOH (800 mL), H2O (400 mL) and THF (800 mL) was heated to 80°C for 2 hr. The mixture was concentrated to dryness, and hot THF (5.0 L) was added to the residue. The mixture was filtered and the filtrate was concentrated. EtOH (300 mL) was added to the residue and stirred for 1 hr. The precipitate was collected by filtration to yield methyl 1-(2-amino-5-bromobenzyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (106 g, 242 mmol, 85.54% yield) as a yellow solid. 1H NMR (400 MHz, 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). ESI [M+H] = 410.2 / 412.2

[0295]

[0296] 4-(7-Bromo-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazin-2-yl)benzonitrile, S10. To a suspension of methyl 1-(2-amino-5-bromobenzyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (105 g, 256 mmol, 1.0 eq.) in toluene (1.5 L) was added MeAl (2 M in toluene, 500 mL, 3.9 eq.) at 0°C and the mixture was stirred at 20°C for 10 hr. The mixture was poured into 1 M glacial HCl (300 mL) and extracted with hot EtOAc / THF (1:1, 500 mL*4). The organic layer was dried over anhydrous NaSO, filtered, and concentrated. EtOH (100 mL) was added to the residue and stirred for 1 hr. The precipitate was collected by filtration to yield 4-(7-bromo-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (86 g, 166 mmol, 65% yield) as a white solid. 1H NMR (400 MHz, 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). ESI [M+H] = 378.2 / 380.2

[0297]

[0298] 4-(7-Bromo-11-chloro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile, S11. A solution of 4-(7-bromo-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (6.2 g, 16 mol, 1.0 eq.) in POCl (100 mL) was stirred for 2 hr at 100° C. The solution was concentrated under reduced pressure to give crude 4-(7-bromo-11-chloro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (16 g, crude) as a dark brown oil, which was used in the next step without further purification.

[0299]

[0300] 4-(7-Bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, S12. To a solution of 4-(7-bromo-11-chloro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (2 g, 4.4 mmol, 1 eq.) in THF / dioxane (1:1, 40 mL) was added TEA to adjust the pH to 7. Formic hydrazide (8 g, 133 mmol, 30.0 eq.) was then added and the mixture was heated to 120° C. in a sealed tube for 10 hr. The mixture was concentrated to dryness and the residue was dissolved in hot EtOAc / THF (1:1, 2.0 L). The solution was washed with 1M HCl (100 mL x 2), saturated aqueous NaHCO₃ (100 mL), and brine (100 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered, and concentrated in vacuo. MeOH (10 mL) was added to the residue and stirred for 1 hr. The precipitate was collected by filtration to yield 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopentan-12-yl)benzonitrile (900 mg, 2.2 mmol, 50.4% yield) as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ = 9.25 (s, 1H), 7.97 (br.s., 1H), 7.82-7.76 (m, 6H), 7.70 (d, J = 8.5Hz, 1H), 7.28 (s, 1H), 5.33 (s, 2H). ESI[M+H]=402.1 / 404.1

[0301] General Procedure A—Ullmann Reaction

[0302]

[0303] 4-(12-(4-cyanophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)piperazine-1-carboxylic acid tert-butyl ester, S13. 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile (30.0 mg, 74 μmol, A mixture of 1,2-dimethyl-1,4-dihydro-1,4-dihydro-2 ...2,4-dihydro-1,4-dihydro-2,4-dihydro-2,4-dihydro-1,4-dihydro-2,4-dihydro-2,4-dihydro-1,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-1,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-1,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-2 The filtrate was washed with brine (5 mL), dried over anhydrous Na2SO4, filtered and concentrated to give tert-butyl 4-(12-(4-cyanophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)piperazine-1-carboxylate (30 mg, crude) as a yellow solid, which was used in the next step without further purification. ESI [M+H] = 508.2

[0304]

[0305] 4-(7-(Piperazin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 1. To a solution of tert-butyl 4-(12-(4-cyanophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)piperazine-1-carboxylate (30 mg, 59.1 umol, 1.0 eq.) in CH2Cl2 (2 mL) was added CF3COOH (2 mL) and the mixture was stirred at 40°C for 30 min. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give 4-(7-(piperazin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile as a yellow solid (15 mg, 27.9 umol, 47.2% yield). 1HNMR (400MHz, DMSO-d6) δ = 9.13 (s, 1H), 8.85 (br.s., 2H), 7.80-7.68 (m, 5H), 7.57 (d, J = 8.8Hz, 1H), 7.2 4(dd,J=2.0,17.2Hz,2H),7.14(dd,J=2.4,8.8Hz,1H),5.21(s,2H),3.23(br.s.,4H),2.52(br.s.,4H). ESI[M+H]=408.1

[0306] Option 3

[0307]

[0308] General Procedure C

[0309]

[0310] tert-Butyl ((1-(12-(4-cyanophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)azetidin-3-yl)methyl)(methyl)carbamate, S16. To 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile (25 mg, 62.1 To a solution of 1,4-dihydro-1,4-dimethyl-2-nitropropane (1,2-dextrin, 1,4 ... The filtrate was concentrated and purified by preparative TLC (SiO 2 , DCM:MeOH=20:1) to give tert-butyl ((1-(12-(4-cyanophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)azetidin-3-yl)methyl)(methyl)carbamate (30 mg, 34.5 umol, 55.5% yield, 60% purity) as a yellow oil, which was used in the next step without further purification. ESI [M+H]=522.3

[0311]

[0312] 4-(7-(3-((methylamino)methyl)azetidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 11. tert-butyl ((1-(12-(4-cyanophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)azetidin-3-yl)methyl)(methyl)carbamate (30.0 mg) A solution of 4-(7-(3-((methylamino)methyl)azetidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile (30.00 mg, crude, TFA) was stirred at 50° C. for 0.5 hr and then concentrated under reduced pressure to give crude 4-(7-(3-((methylamino)methyl)azetidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile as a yellow oil (30.00 mg, crude, TFA) which was used in the next step without further purification. ESI [M+H] = 422.1.

[0313]

[0314] To a solution of 4-(7-(3-((dimethylamino)methyl)azetidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile (30.0 mg, 71.17 umol, 1.0 eq.) in MeOH (2.0 mL) was added HCHO (57.7 mg, 711.7 umol, 10.0 eq.) and the mixture was stirred at 15 °C for 2 hr. NaBH3CN (6.71 mg, 106.7 umol, 1.5 eq.) was then added and the mixture was stirred for 14 hr at 15° C. The mixture was concentrated under reduced pressure and purified by preparative HPLC to yield 4-(7-(3-((dimethylamino)methyl)azetidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile (6.0 mg, 13.6 umol, 19.1% yield, 99% purity) as a white solid. 1HNMR (400MHz, methanol-d4) δ=9.12(br.s.,1H),7.78-7.62(m,5H),7.51(d,J=8.7Hz,1H),7.25(br.s.,1H),6.71(d,J=2.4Hz,1H),6.61( dd,J=2.4,8.7Hz,1H),5.17(s,2H),4.21(t,J=7.8Hz,2H),3.82-3.74(m,2H),3.54(d,J=7.3Hz,2H),3.29-3.23(m,1H),2.95(s,6H). ESI[M+H]=436.2

[0315] Option 5

[0316]

[0317] General Procedure E

[0318]

[0319] 4-(7-(2-((dimethylamino)methyl)morpholinyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, S18. Synthesized using General Procedure A with N,N-dimethyl-1-(morpholin-2-yl)methanamine replacing tert-butyl piperazine-1-carboxylate. ESI [M+H] = 466.2

[0320]

[0321] (S)-4-(7-(2-((dimethylamino)methyl)morpholinyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 23. 4-(7-(2-((dimethylamino)methyl)morpholinyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile was separated by SFC to yield (S)-4-(7-(2-((dimethylamino)methyl)morpholinyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile as a white solid. 1H NMR (400 MHz, methanol-d4) δ = 9.01 (s, 1H), 7.74-7.68 (m, 2H), 7.68-7.63 (m, 2H), 7.61 (s, 1H), 7.54 (d, J = 8.8 Hz, 1H), 7.21 (d, J = 4.4 Hz, 2H), 7.10 (dd, J = 2.2, 8.8 Hz, 1H), 5.18 (s, 2H), 4.10 (d, J = 9 .7Hz,1H),4.00(t,J=9.3Hz,1H),3.87-3.78(m,1H),3.72(d,J=11.5Hz,1H),3.64(d,J=11.9H z,1H),3.17-3.01(m,2H),2.90(dt,J=3.1,11.9Hz,1H),2.76(s,6H),2.59(t,J=11.0Hz,1H). ESI[M+H]=466.2

[0322] Option 7

[0323]

[0324] General Procedure G

[0325]

[0326] Benzyl ((trans-1-(12-(4-cyanophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)-4-hydroxypyrrolidin-3-yl)methyl)(methyl)carbamate, S22. Synthesized using general procedure A substituting benzyl ((trans-4-hydroxypyrrolidin-3-yl)methyl)(methyl)carbamate for tert-butyl piperazine-1-carboxylate. ESI [M+H] = 586.2

[0327]

[0328] To a solution of benzyl ((trans-1-(12-(4-cyanophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-7-yl)-4-hydroxypyrrolidin-3-yl)methyl)(methyl)carbamate (30 mg, 20 umol, 1.0 eq.) in DCM (2.0 mL) was added TMSI (20.5 mg, 102.4 umol, 14 uL, 5.0 eq.). The mixture was stirred at 26° C. for 0.5 hr and then concentrated. The residue was purified by acidic preparative HPLC to give 4-(7-(trans-3-hydroxy-4-((methylamino)methyl)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopentan-12-yl)benzonitrile as a yellow solid (1.93 mg, 3.28 umol, 16.% yield). 1 H NMR (400MHz, methanol-d4) δ=9.00(s,1H),7.77-7.70(m,2H),7.70-7.64(m,2H),7.62(s,1H),7.50(d,J=8.8Hz,1H),7.23(s,1H),6.80(d,J=2.2Hz ,1H),6.70(dd,J=2.6,8.8Hz,1H),5.18(s,2H),4.30(q,J=6.6Hz,1H),3.78-3.64(m,2H),3.27-3.10(m,4H),2.77(s,3H),2.62-2.52(m,1H). ESI[M+H]=452.1

[0329] Option 8

[0330]

[0331] General Procedure H

[0332]

[0333] 4-(7-(4-Hydroxypiperidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, S23. Synthesized using general procedure A with piperidin-4-ol replacing tert-butyl piperazine-1-carboxylate. ESI [M+H] = 423.2

[0334]

[0335] 4-(7-(4-Oxopiperidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, S24. To a solution of 4-(7-(4-hydroxypiperidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile (690 mg, 1.6 mmol, 1.0 eq.) in DCM (20 mL) was added Dess-Martin (898 mg, 2.1 mmol, 1.3 eq.) in one portion and the mixture was stirred at 20° C. for 16 hr. After completion of the reaction, the mixture was filtered. The filtrate was concentrated and the residue was purified by silica gel chromatography (DCM:MeOH=20:1, Rf=0.50) to give 4-(7-(4-oxopiperidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile (600 mg, 78.8% yield, 90% purity) as a yellow solid. 1HNMR (400MHz, DMSO-d6) δ = 9.14-9.07 (m, 1H), 7.80-7.68 (m, 5H), 7.57-7.49 (m, 1H), 7.27 (d, J = 2.6Hz, 1H), 7. 20(d,J=1.8Hz,1H),7.13(dd,J=2.6,8.8Hz,1H),5.25-5.14(m,2H),3.68(t,J=5.8Hz,4H),2.44-2.34(m,4H). ESI[M+H]=421.0

[0336]

[0337] (S)-4-(7-(4-((1-hydroxypropyl-2-yl)amino)piperidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine-12-yl)benzonitrile, S25. 4-(7-(4-oxopiperidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine-12-yl)benzonitrile A mixture of nitrile (40 mg, 95 umol, 1.0 eq.) and (2S)-2-aminopropan-1-ol (7.1 mg, 95.1 umol, 1.0 eq.) in MeOH (3.0 mL) was stirred at 25° C. for 2 hr, followed by the addition of Ti(i-PrO) 4 (5.4 mg, 19.0 umol, 5.6 uL, 0.2 eq.) and NaBH 3 CN (11.9 mg, 190.2 umol, 2.0 eq.). The mixture was stirred at 25° C. for 10 hr and concentrated. The residue was purified by preparative HPLC (TFA conditions) to give (S)-4-(7-(4-((1-hydroxypropan-2-yl)amino)piperidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopentan-12-yl)benzonitrile as a white solid (15.8 mg, 26.7 umol, 28% yield, 100% purity, TFA salt). 1H NMR (400MHz, methanol-d4) δ = 9.04 (s, 1H), 7.74-7.57 (m, 5H), 7.51 (d, J = 8.8Hz, 1H ),7.25-7.18(m,2H),7.11(dd,J=2.2,8.8Hz,1H),5.16(s,2H),4.00(d,J=1 2.8Hz,2H),3.82(dd,J=3.5,11.9Hz,1H),3.64-3.41(m,3H),2.94(t,J=12. 1Hz, 2H), 2.18 (d, J = 11.5Hz, 2H), 1.90-1.65 (m, 2H), 1.33 (d, J = 6.6Hz, 3H). ESI[M+H]=480.2

[0338]

[0339] (S)-4-(7-(4-((1-hydroxypropan-2-yl)(methyl)amino)piperidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 104. (S)-4-(7-(4-((1-hydroxypropan-2-yl)amino)piperidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile A mixture of triazolo[3,4-c][1,4]diazopentan-12-yl)benzonitrile (20 mg, 41.7 umol, 1.0 eq.) and formaldehyde (16.9 mg, 208.5 umol, 15.5 uL, 5.0 eq.) in MeOH (2.0 mL) was stirred at 30 °C for 1 hr, then NaBH3CN (5.2 mg, 83.4 umol, 2.0 eq.) was added and the mixture was stirred for 1 hr. The mixture was concentrated and the residue was purified by preparative HPLC (TFA conditions) to give (S)-4-(7-(4-((1-hydroxypropan-2-yl)(methyl)amino)piperidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazol-12-yl)benzonitrile (4.5 mg, 7.2 umol, 17.4% yield, 98% purity, TFA) as a white solid. NMR (400 MHz, methanol-d4) δ = 9.04 (d, J = 3.5 Hz, 1H), 7.76-7.59 (m, 5H), 7.53 (d, J = 8.8 Hz, 1H), 7.26-7.20 (m, 2H), 7.12 (d, J = 9.3 Hz, 1H), 5.19 (s, 2H), 4.04 (d, J = 12.3 Hz, 2H), 3.83 -3.74(m,2H),3.63(d,J=4.0Hz,2H),2.92(t,J=12.6Hz,2H),2.84(s,1H),2.76(s,2H ),2.34-2.13(m,2H),1.99-1.75(m,2H),1.37(d,J=6.2Hz,1H),1.27(d,J=6.6Hz,2H). ESI[M+H]=494.2

[0340] Option 9

[0341]

[0342] Chemical experimental methods:

[0343]

[0344]

[0266] 4-(7-Bromo-11-((2,2-dimethoxyethyl)amino)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile, S26. To a solution of 4-(7-bromo-11-chloro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (2.1 g, 5.2 mmol, 1.0 eq.) in THF (20 mL) and dioxane (40 mL) was added 2,2-dimethoxyethylamine (8.8 g, 83.8 mmol, 15 eq.) and the mixture was heated at 100 °C in a sealed tube for 16 hr. The resulting mixture was concentrated and then partitioned between EtOAc (1000 mL) and 0.5 M HCl (150 mL). The organic layer was dried and concentrated to afford 4-(7-bromo-11-((2,2-dimethoxyethyl)amino)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (1.6 g) as a brown solid. 1H NMR (400 MHz, DMSO-d6) δ = 11.51 (s, 1H), 10.15 (s, 1H), 8.01-7.27 (m, 9H), 5.38 (s, 2H), 4.77 (s, 1H), 3.98-3.90 (s, 2H), 3.57-3.39 (s, 6H). ESI [M+H] = 464.9 / 466.9

[0345]

[0346]

[0266] 4-(7-Bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, S27. A mixture of 4-(7-bromo-11-((2,2-dimethoxyethyl)amino)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (1.6 g, 3.4 mmol, 1.0 eq.) in 1 M HCl (60 mL) and dioxane (60 mL) was stirred at 60 °C for 20 hr. The reaction mixture was concentrated to give 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (1.4 g, crude) as a gray solid, which was used in the next step without further purification. 1H NMR (400 MHz, DMSO-d6) δ = 8.30 (s, 1H), 8.01 (s, 1H), 7.93 (s, 2H), 7.86-7.83 (m, 4H), 7.76-7.70 (m, 2H), 7.46 (s, 1H), 5.47 (s, 2H). ESI [M+H] = 401.0 / 402.9

[0347]

[0348] 4-(7-(2-methyl-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, 125. The reaction mixture was prepared using General Procedure C with 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4] The obtained product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile with tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate with tert-butyl (azetidin-3-ylmethyl)(methyl)carbamate. 1H NMR (400MHz, methanol-d4) δ = 7.99 (d, J = 1.8Hz, 1H), 7.78-7.67 (m, 6H), 7.54 (d, J = 8.8Hz, 1H), 7.28 (s, 1H), 6.84-6.72 (m, 2H), 5. 29(s,2H),4.40-4.24(m,2H),4.20-4.06(m,2H),3.71-3.59(m,2H),3.46(br.s.,2H),2.98(br.s.,3H),2.40(br.s.,2H). ESI[M+H]=447.1

[0349] Plan 10

[0350]

[0351] General Procedure I

[0352]

[0353]

[0266] 4-(7-Bromo-11-chloro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, S29. To a solution of 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile (1.0 g, 2.5 mmol, 1.0 eq.) in THF (20 mL) and DMF (30 mL) was added NCS (399 mg, 3 mmol, 1.2 eq.) and the mixture was stirred at 25 °C for 16 hr. The reaction mixture was concentrated and purified by column chromatography (SiO2, petroleum ether / ethyl acetate / THF = 5 / 1 / 1 to 1 / 1 / 1) to give 4-(7-bromo-11-chloro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopyridine-12-yl)benzonitrile (760 mg, 1.7 mmol, 69.8% yield) as a gray solid. ESI [M+H] = 435.9 / 437.9

[0354]

[0355] (S)-tert-Butyl((1-(11-chloro-12-(4-cyanophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazol-7-yl)pyrrolidin-3-yl)methyl)carbamate, S30.

[0356] Synthesized using General Procedure A with 4-(7-bromo-11-chloro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl (R)-(pyrrolidin-3-ylmethyl)carbamate replacing tert-butyl piperazine-1-carboxylate. ESI [M+H] = 556.4

[0357]

[0358] (R)-((1-(11-chloro-12-(4-cyanophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazol-7-yl)pyrrolidin-3-yl)methyl)(methyl)carbamic acid tert-butyl ester, S31. To (S)-((1-(11-chloro-12-(4-cyanophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazol-7-yl)pyrrolidin-3-yl)methyl)(methyl)carbamate To a solution of tert-butyl (4-c) [1,4-diazopyridin-7-yl)pyrrolidin-3-yl)methyl)carbamate (15 mg, 26.9 μmol, 1.0 eq.) and MeI (11.4 mg, 80.9 μmol, 5.04 μL, 3.0 eq.) in DMF (2.0 mL) was added NaH (2.1 mg, 53.9 μmol, 60% purity, 2.0 eq.), and the mixture was stirred at 25° C. for 20 min. The mixture was then poured into water (10 mL) and extracted with EtOAc (30 mL*3). The organic layer was washed with brine (10 mL), dried over MgSO4, filtered and concentrated to give crude (R)-tert-butyl((1-(11-chloro-12-(4-cyanophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-7-yl)pyrrolidin-3-yl)methyl)(methyl)carbamate as a brown oil (20 mg), which was used in the next step without further purification. ESI [M+H] = 570.1

[0359]

[0360]

[0147] A solution of (S)-4-(11-chloro-7-(3-((methylamino)methyl)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile, 173. A solution of (R)-tert-butyl((1-(11-chloro-12-(4-cyanophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-7-yl)pyrrolidin-3-yl)methyl)(methyl)carbamate (20 mg, 35 umol, 1.0 eq.) in TFA (2.0 mL) was stirred at 50 °C for 5 min. The reaction was concentrated and purified by preparative HPLC (TFA conditions) to give (S)-4-(11-chloro-7-(3-((methylamino)methyl)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazol-12-yl)benzonitrile (2.31 mg, 3.83 umol, 10.91% yield, 96.8% purity, TFA salt) as a yellow solid. NMR (400 MHz, methanol-d4) δ = 9.03 (br.s., 1H), 7.87-7.79 (m, 2H), 7.78-7.70 (m, 2H), 7.51 (d, J = 8.8 Hz, 1H), 7.13 (s, 1H), 6.80 (d, J = 2.2 Hz, 1H), 6.73 (dd, J = 2.4, 8.8 Hz, 1H), 5.25 (br .s.,2H),3.68-3.58(m,1H),3.53(dt,J=3.9,8.9Hz,1H),3.48-3.38(m,1H),3.24-3.10 (m,3H),2.82-2.66(m,4H),2.33(dd,J=4.5,11.4Hz,1H),1.89(qd,J=8.4,12.4Hz,1H). ESI[M+H]=470.1

[0361] Plan 11

[0362]

[0363] Chemical experimental methods:

[0364]

[0365] 4-(7-Bromo-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, S32. To a solution of 4-(7-bromo-11-chloro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (2.1 g, 5.3 mmol, 1.0 eq.) in toluene / THF (5:2, 70 mL) was added prop-2-yn-1-amine (5 g, 91.1 mmol, 17.2 eq.) and the mixture was heated in a sealed tube at 120° C. for 56 hr. The resulting mixture was concentrated and partitioned between THF / EtOAc (1:1, 200 mL) and 0.25 M glacial HCl (100 mL). The separated aqueous layer was extracted with THF / EtOAc (1:1, 100 mL x 3). The combined organic layers were washed with brine (100 mL), dried over anhydrous MgSO₄, filtered, and concentrated in vacuo. The residue was purified by column chromatography to yield 4-(7-bromo-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopentan-12-yl)benzonitrile (1.8 g, 3.0 mmol, 57% yield) as a gray solid. 1H NMR (400 MHz, chloroform-d) δ = 7.63 (s, 1H), 7.63-7.51 (m, 5H), 7.20 (d, J = 8.4 Hz, 1H), 7.13 (d, J = 1.6 Hz, 1H), 7.07 (s, 1H), 7.01 (s, 1H), 5.07 (d, 14.0 Hz, 1H), 4.81 (d, 14.2 Hz, 1H), 2.38 (s, 3H). ESI [M+H] = 415.2 / 417.2

[0366]

[0367] 4-(3-Methyl-7-(2,6-diazaspiro[3.4]octan-6-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile, 201. Synthesized using general procedure A substituting 4-(7-bromo-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile and tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ=7.81(s,1H),7.78-7.72(m,4H),7.59(s,1H),7.50(d,J=8.8Hz,1H),7.28(s,1H),6.88(br.s.,1H),6.77(d,J=8 .8Hz,1H),5.26(s,2H),4.23-4.17(m,2H),4.15-4.08(m,2H),3.67(s,2H),3.50(t,J=6.8Hz,2H),2.53(s,3H),2.43(t,J=6.8Hz,2H). ESI[M+H]=447.1

[0368] Plan 12

[0369]

[0370] Chemical experimental methods:

[0371]

[0372] 4-(7-Bromo-11-oxo-10-(2-oxopropyl)-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazin-2-yl)benzonitrile, S34. To a mixture of 4-(7-bromo-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazin-2-yl)benzonitrile (2.5 g, 6.6 mmol, 1.0 eq.), 18-crown-6 (192 mg, 727 umol, 0.11 eq.) and K2CO3 (5.0 g, 36.1 mmol, 5.4 eq.) in DMF (40 mL) was added 1-chloropropan-2-one (12 g, 129.7 mmol, 19.6 eq.) at 15 °C. The reaction mixture was heated to 70° C. (oil bath) and stirred for 4 days. The reaction mixture was concentrated in vacuo, diluted with EtOAc (100 mL), washed with water (10 mL×2) and brine (50 mL), dried over Na 2 SO 4 , and then concentrated to yield 4-(7-bromo-11-oxo-10-(2-oxopropyl)-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-2-yl)benzonitrile (3.0 g, crude) as a dark brown oil, which was used directly without purification. ESI[M+H]=434.0 / 436.0

[0373]

[0374]

[0147] 4-(7-Bromo-2-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazin-12-yl)benzonitrile, S35. A mixture of 4-(7-bromo-11-oxo-10-(2-oxopropyl)-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazin-2-yl)benzonitrile (2.5 g, 5.7 mmol, 1.0 eq.) and NH4OAc (25 g, 324 mmol, 56.3 eq.) in AcOH (200 mL) was stirred at 120°C for 48 hr. The mixture was concentrated and the residue was purified by preparative HPLC (TFA) to give 4-(7-bromo-2-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (450 mg, 845 umol, 14.68% yield, 78% purity) as a brown solid, which was used directly. ESI [M+H] = 414.8 / 416.8

[0375]

[0376]

[0147] 4-(7-(cis-3,5-dimethylpiperazin-1-yl)-2-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 215. Synthesized using general procedure A, substituting 4-(7-bromo-2-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and cis-2,6-dimethylpiperazine for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 7.83-7.69 (m, 6H), 7.62 (d, J = 8.8Hz, 1H), 7.32 (dd, J = 2.2, 11.9Hz, 2H), 7.26 (dd, J = 2.6, 8.8Hz, 1H), 5 .35(s,2H),4.08(d,J=11.5Hz,2H),3.51(d,J=6.8Hz,2H),2.82(dd,J=11.6,13.3Hz,2H),2.49(s,3H),1.42(d,J=6.4Hz,6H). ESI[M+H]=449.2

[0377] Plan 13

[0378]

[0379] Chemical experimental methods:

[0380]

[0381] 4-(7-Bromo-11-oxo-10-(3-oxobutan-2-yl)-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile, S36. To a solution of 4-(7-bromo-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (1.0 g, 2.6 mmol, 1.0 eq.) and 3-chlorobutan-2-one (560 mg, 2.0 eq.) in DMF (20 mL) were added KCO (1.8 g, 13.2 mmol, 5.0 eq.) and 18-C-6 (348 mg, 1.3 mmol, 0.5 eq.). The mixture was stirred at 70°C for 12 hours and poured into ice water (100 mL). The mixture was extracted with ethyl acetate (200 mL*3), washed with brine (30 mL*2), dried over anhydrous Na2SO4, filtered and concentrated to produce 4-(7-bromo-11-oxo-10-(3-oxobutan-2-yl)-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (1.5 g, crude) as a dark brown oil. ESI[M+H]=447.9 / 449.9

[0382]

[0383] 4-(7-Bromo-2,3-dimethyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazin-12-yl)benzonitrile, S37. A mixture of 4-(7-bromo-11-oxo-10-(3-oxobutan-2-yl)-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazin-2-yl)benzonitrile (1.5 g, 3.3 mmol, 1.0 eq.) and NHOAc (7.7 g, 100 mmol, 30.0 eq.) in HOAc (50 mL) was heated to 120° C. under N atmosphere for 16 hr and then concentrated under reduced pressure. The residue was poured into ice water (50 mL) and extracted with ethyl acetate / THF (1:1, 30 mL*5). The combined organic phases were washed with brine (30 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by preparative HPLC (TFA conditions) to yield 4-(7-bromo-2,3-dimethyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine-12-yl)benzonitrile (700 mg, 1.3 mmol, 38.94% yield, 80% purity) as a brown oil. ESI [M+H] = 429.2 / 431.2

[0384]

[0385] 4-(7-(cis-3,5-dimethylpiperazin-1-yl)-2,3-dimethyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 221. Synthesized using General Procedure A substituting 4-(7-bromo-2,3-dimethyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c]-[1,4]diazepin-12-yl)benzonitrile and cis-2,6-dimethylpiperazine for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 7.77-7.66 (m, 5H), 7.52 (d, J = 8.8Hz, 1H), 7.35 (br.s., 1H), 7.28-7.17 (m, 2H), 5.26 (s, 2H), 4.07 (d, J = 13.2Hz, 2H), 3.50 (br.s., 2H), 2.87-2.76 (m, 2H), 2.42 (d, J = 4.4Hz, 6H), 1.41 (d, J = 6.2Hz, 6H). ESI[M+H]=463.2

[0386] Plan 14

[0387]

[0388] Chemical experimental methods:

[0389]

[0390] 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, S38. A solution of 4-(7-bromo-11-chloro-5H-benzo[e]-pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (2.0 g, 5 mmol, 1.0 eq.) and acetic acid hydrazide (746 mg, 10 mmol, 2.0 eq.) in dioxane (25 mL) was stirred at 110° C. for 16 hr. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with ice water (20 mL) and extracted with hot ethyl acetate / THF (2:1, 25 mL*3). The combined organic phases were washed with brine (30 mL), dried over anhydrous NaSO, filtered, and concentrated in vacuo to afford a dark brown solid. The solid was washed with MeOH (10 mL) to yield 4-(7-bromo-3-methyl-9H-benzo[e]pyrrolo[1,2-a]-[1,2,4]triazolo-[3,4-c][1,4]-diazopyridine-12-yl)benzonitrile (1.30 g, crude) as a light yellow solid. 1H NMR (400 MHz, DMSO-d6) δ = 7.97 (s, 1H), 7.80-7.50 (m, 6H), 7.70-7.64 (m, 1H), 7.19 (s, 1H), 5.34-5.13 (m, 2H), 2.58 (s, 3H).

[0391]

[0392] 4-(7-((S)-3-aminopyrrolidin-1-yl)-3-methyl-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 230. The general procedure A was used with 4-(7-bromo-3-methyl-9H-benzo[e]pyrrolo-[1,2-a][1,2,4 The present invention was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c]-[1,4]diazopyridine-12-yl)benzonitrile with tert-butyl (S)-pyrrolidin-3-ylcarbamate in place of tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 7.71-7.62 (m, 5H), 7.46 (d, J = 8.8Hz, 1H), 7.23 (br.s., 1H), 6.89 (br.s., 1H), 6.75 (d, J = 8.8Hz, 1H), 5.1 7(br.s.,2H),4.07(br.s.,1H),3.75-3.60(m,2H),3.54-3.43(m,2H),2.70(br.s.,3H),2.55-2.44(m,1H),2.25-2.16(m,1H). ESI[M+H]=422.1

[0393] Plan 15

[0394]

[0395] Chemical experimental methods:

[0396]

[0397] 4-(7-Bromo-9H-benzo[e]pyrrolo[1,2-a]tetrazolo[5,1-c][1,4]diazepin-12-yl)benzonitrile, S40. A mixture of 4-(7-bromo-11-chloro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (850 mg, 2.1 mmol, 1.0 eq.) and TMSN3 (271.5 mg, 2.3 mmol, 1.1 eq.) in DMF (10 mL) was stirred at 10° C. for 16 hr and then concentrated. MeOH (20 mL) was added to the residue and stirred for 1 hr. The precipitate was collected by filtration to give 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a]tetrazolo[5,1-c][1,4]diazepin-12-yl)benzonitrile (600 mg, 62.5% yield, 90% purity) as a red solid. ESI [M+H] = 403.0 / 405.0

[0398]

[0399] 4-(7-(cis-3,5-dimethylpiperazin-1-yl)-9H-benzo[e]pyrrolo[1,2-a]tetrazolo[5,1-c][1,4]diazepin-12-yl)benzonitrile, 242. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a]tetrazolo[5,1-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and cis-2,6-dimethylpiperazine for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 7.89 (d, J = 8.8Hz, 1H), 7.78-7.74 (m, 2H), 7.71-7.68 (m, 3H), 7.40 (d, J = 1.8Hz, 1H), 7.34-7 .24(m,2H),5.30(s,2H),4.09(d,J=11.5Hz,2H),3.52(d,J=7.1Hz,2H),2.86-2.79(m,2H),1.42(d,J=6.6Hz,6H). ESI[M+H]=437.2

[0400] Plan 16

[0401]

[0402] Chemical experimental methods:

[0403]

[0404] 4-(10-Amino-7-bromo-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile, S41. To a solution of 4-(7-bromo-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (1 g, 2.64 mmol, 1.0 eq.) in DMF (20 mL) was added NaH (169 mg, 4.2 mmol, 60% purity, 1.6 eq.) at 0° C. After stirring at 0° C. for 1 hr, (aminooxy)diphenylphosphine oxide (738 mg, 3.2 mmol, 1.2 eq.) was added and the mixture was stirred at 20° C. for 1 hr. 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. MTBE (20 mL) was added to the residue and stirred for 1 hr. The precipitate was collected by filtration to produce 4-(10-amino-7-bromo-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-2-yl)benzonitrile (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

[0405]

[0406] 4-(7-Bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazepin-12-yl)benzonitrile, S42. A solution of 4-(10-amino-7-bromo-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (900 mg, 2.29 mmol, 1.0 eq.) in formamide (10 mL) was stirred at 200° C. for 1.5 hr, then the mixture was poured into ice water (20 mL) and filtered. EtOH (10 mL) was added to the filter cake and stirred for 1 hr. The precipitate was collected by filtration to yield 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazopyridine-12-yl)benzonitrile (600 mg) as a yellow solid. 1H NMR (400 MHz, 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). ESI [M+H] = 402.1 / 404.1

[0407]

[0408] 4-(7-(2,6-diazaspiro[3.4]octan-6-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazepin-12-yl)benzonitrile, 256. The general procedure A was used with 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5 The product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile with tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate instead of tert-butyl hexahydropyrazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 8.16 (s, 1H), 7.79-7.64 (m, 5H), 7.62 (s, 1H), 7.21 (s, 1H), 6.80-6.70 (m, 2H), 5.19 (s,2H),4.21-4.13(m,2H),4.12-4.04(m,2H),3.63(s,2H),3.47(t,J=6.8Hz,2H),2.39(t,J=6.9Hz,2H). ESI[M+H]=434.1

[0409] Plan 19

[0410]

[0411] Chemical experimental methods:

[0412]

[0413] 4-(11-Amino-7-bromo-5H-benzo[e]pyrrolo[1,2-a][1,4]diazin-2-yl)benzonitrile, S49. To a solution of 4-(7-bromo-11-chloro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazin-2-yl)benzonitrile (1.6 g, 4 mmol, 1.0 eq.) in MeCN (20 mL) and THF (20 mL) was added NH3.HO (1.4 g, 40 mmol, 10.0 eq.) and the mixture was heated at 120 °C in a sealed tube for 16 hr. The reaction mixture was concentrated to give 4-(11-amino-7-bromo-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (1.4 g, crude) as a yellow solid, which was used directly in the next step without further purification. ESI [M+H] = 377.1 / 379.1

[0414]

[0415] 4-(7-Bromo-3-formyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, S50. To a solution of 4-(11-amino-7-bromo-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (1.4 g, 3.7 mmol, 1.0 eq.) in i-PrOH (10 mL) was added EtN (413 mg, 4 mmol, 1.1 eq.) and 2-bromomalonaldehyde (616 mg, 4 mmol, 1.0 eq.). The mixture was stirred at 10° C. for 0.1 hr and then AcOH (267 mg, 4.4 mmol, 1.2 eq.) was added. The mixture was heated to 90° C. for 2 hr and concentrated. The residue was purified by preparative HPLC (TFA conditions) to give 4-(7-bromo-3-formyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (300 mg, 545 umol, 14.68% yield) as a yellow solid. 1H NMR (400 MHz, DMSO-d6) δ = 9.71 (s., 1H), 8.24 (s, 1H), 7.88 (s, 1H), 7.78 (s, 1H), 7.75 (s, 4H), 7.62 (d, J = 12 Hz, 1H), 7.34 (d, J = 12 Hz, 1H), 7.29 (s, 1H), 5.35 (d, J = 12 Hz, 1H), 5.22 (d, J = 12 Hz, 1H). ESI[M+H]=429.0 / 431.0

[0416]

[0417] 4-(7-Bromo-3-(hydroxymethyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, S51. To a solution of 4-(7-bromo-3-formyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (300 mg, 699 umol, 1.0 eq.) in MeOH (30 mL) was added NaBH4 (105 mg, 2.8 mmol, 4.0 eq.) and the mixture was stirred at 10°C for 16 hr. The reaction mixture was concentrated, diluted with water (30 mL) and extracted with EtOAc (30 mL*3). The organic phase was concentrated to give 4-(7-bromo-3-(hydroxymethyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (300 mg, 695.6 umol, 99.53% yield) as a light yellow solid, which was used directly in the next step without further purification. ESI [M+H] = 431.1 / 433.1

[0418]

[0419] 4-(7-((R)-3-(Dimethylamino)pyrrolidin-1-yl)-3-(hydroxymethyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 274. Synthesized using general procedure A substituting 4-(7-bromo-3-(hydroxymethyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (R)-N,N-dimethylpyrrolidin-3-amine for tert-butyl piperazine-1-carboxylate. 1H NMR (400 MHz, methanol-d4) δ = 7.82 (d, J = 8.8 Hz, 1H), 7.79-7.75 (m, 2H), 7.74-7.70 (m, 2H), 7.70-7.65 (m, 2H), 7.28 (d, J = 1.8 Hz, 1H), 6.92 (d, J = 2.2 Hz, 1H), 6.80 (dd, J = 2.4, 9.0 Hz, 1H), 5.24 (d, J = 7.1 Hz, 2H), 4.81 (d,J=14.1Hz,1H),4.62(d,J=14.1Hz,1H),4.07(quin,J=7.2Hz,1H),3.82(ddd,J=3.7,7.2,10.7Hz,1 H), 3.73-3.57 (m, 2H), 3.45 (q, J = 7.5Hz, 1H), 3.00-2.94 (m, 6H), 2.65-2.54 (m, 1H), 2.39-2.27 (m, 1H). ESI[M+H]=465.1

[0420] Plan 20

[0421]

[0422] Chemical experimental methods:

[0423]

[0424]

[0266] 4-(7-Bromo-3-((methylamino)methyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, S52. To a solution of 4-(7-bromo-3-formyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (100 mg, 232 umol, 1.0 eq.) in MeOH (5.0 mL) was added MeNH (144 mg, 4.6 mmol, 20 eq.) and the mixture was stirred at 10°C for 1 hr. NaBH3CN (58.5 mg, 931.8 umol, 4.0 eq.) was then added, and the mixture was stirred at 10°C for 16 hours and concentrated to yield 4-(7-bromo-3-((methylamino)methyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (110 mg, crude) as a dark brown solid, which was used directly in the next step without further purification. ESI [M+H] = 444.0 / 446.0

[0425]

[0426] To a solution of 4-(7-bromo-3-((methylamino)methyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile (110 mg, 247 umol, 1.0 eq.) in THF (5 mL) was added (Boc)O (59 mg, 272 umol, 1.1 eq.) and EtN (50.1 mg, 495 umol, 2.0 eq.). The mixture was stirred at 10° C. for 1 hr and concentrated to dryness. The mixture was purified by preparative TLC (SiO 2 , ethyl acetate:methanol=20:1) to give tert-butyl ((7-bromo-12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-3-yl)methyl)(methyl)carbamate (150 mg, crude) as a yellow solid. ESI [M+H]=544.1 / 546.1

[0427]

[0428] 4-(7-((R)-3-(dimethylamino)pyrrolidin-1-yl)-3-((methylamino)methyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 276. The general procedure A was used with ((7-bromo-12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c] The product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile with tert-butyl pyrrolo[1,2-a][1,4]diazon-3-yl)methyl)(methyl)carbamate and tert-butyl piperazine-1-carboxylate with (R)-N,N-dimethylpyrrolidin-3-amine. NMR (400 MHz, methanol-d4) δ = 7.74-7.70 (m, 2H), 7.69-7.64 (m, 4H), 7.30 (d, J = 8.8 Hz, 1H), 7.17 (s, 1H), 6.95 (d, J = 2.2 Hz, 1H), 6.83-6.77 (m, 1H), 5.25-5.16 (m, 2H), 4.67-4.60 (m,1H),4.54-4.45(m,1H),4.14-4.04(m,1H),3.87-3.76(m,1H),3.75-3.60(m,2H) ,3.51-3.40(m,1H),2.99(s,6H),2.68(s,3H),2.66-2.56(m,1H),2.38-2.27(m,1H). ESI[M+H]=478.2

[0429] Plan 21

[0430]

[0431] Chemical experimental methods:

[0432]

[0433]

[0266] 4-(3-(Aminomethyl)-7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, S55. To a solution of 4-(7-bromo-3-formyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (100 mg, 233 umol, 1.0 eq.) in MeOH (5.0 mL) was added NH3H2O (81 mg, 2.3 mmol, 10 eq.) and NHOAc (35.9 mg, 465.9 umol, 2.0 eq.). The mixture was stirred at 80°C for 2 hr, and then NaBH3CN (29 mg, 465.9 umol, 2.0 eq.) was added. The reaction mixture was heated at 80° C. for 16 hours and concentrated to give crude 4-(3-(aminomethyl)-7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (110 mg) as a dark brown solid, which was used directly in the next step without further purification. ESI [M+H] = 430.2 / 432.0

[0434]

[0435] tert-Butyl ((7-bromo-12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-3-yl)methyl)carbamate, S56. To a solution of 4-(3-(aminomethyl)-7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile (110 mg, 255 umol, 1.0 eq.) in THF (5 mL) was added (Boc)O (61.3 mg, 281.2 umol, 1.1 eq.) and EtN (51.7 mg, 511.2 umol, 2.0 eq.). The reaction mixture was stirred at 10° C. for 1 hr, concentrated and analyzed by preparative TLC (SiO 2 , dichloromethane:methanol 10:1, R f =0.75) to give tert-butyl ((7-bromo-12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-3-yl)methyl)carbamate (120 mg) as a yellow solid. ESI [M+H] = 530.1 / 532.1

[0436]

[0437] 4-(3-(Aminomethyl)-7-((R)-3-(dimethylamino)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 277. The reaction mixture was prepared using General Procedure A with ((7-bromo-12-(4-cyanophenyl)-9H-benzo[e]imidazo[2,1-c] The product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile with tert-butyl pyrrolo[1,2-a][1,4]diazon-3-yl)methyl)carbamate and (R)-N,N-dimethylpyrrolidin-3-amine with tert-butyl piperazine-1-carboxylate. NMR (400 MHz, methanol-d4) δ = 7.74-7.68 (m, 2H), 7.68-7.63 (m, 2H), 7.58 (s, 1H), 7.50 (s, 1H), 7.28 (d, J = 8.4 Hz, 1H), 7.10 (s, 1H), 6.95 (s, 1H), 6.79 (d, J = 8.8 Hz, 1H), 5.25-5.06 (m, 2H), 4.53 (d, J =15.0Hz,1H),4.32(d,J=15.4Hz,1H),4.13-4.04(m,1H),3.81(br.s.,1H),3.72-3.69(m,1H) ,3.61(br.s.,1H),3.50-3.39(m,1H),2.99(s,6H),2.66-2.57(m,1H),2.32(d,J=12.8Hz,1H). ESI[M+H]=464.2

[0438] Plan 22

[0439]

[0440] Chemical experimental methods:

[0441]

[0442] 4-(7-Bromo-3-(methoxymethyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, S58. To a solution of 4-(7-bromo-3-(hydroxymethyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (60 mg, 139 umol, 1.0 eq.) in THF (10 mL) was added NaH (11 mg, 278 umol, 2.0 eq.) at 0° C. After stirring for 0.1 hr, MeI (23.7 mg, 167 umol, 1.2 eq.) was added to the reaction mixture and the mixture was stirred at 10° C. for 16 hr. The mixture was quenched with saturated NH4Cl solution (50 mL), extracted with EtOAc (30 mL*2) and concentrated. The results were analyzed by preparative TLC (SiO2, petroleum ether:ethyl acetate=1:1, R f =0.30) to give 4-(7-bromo-3-(methoxymethyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (40 mg) as a yellow solid. ESI [M+H] = 445.0 / 447.0

[0443]

[0444] 4-(7-((R)-3-(Dimethylamino)pyrrolidin-1-yl)-3-(methoxymethyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 278. Synthesized using general procedure A substituting 4-(7-bromo-3-(methoxymethyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (R)-N,N-dimethylpyrrolidin-3-amine for tert-butyl piperazine-1-carboxylate. 1H NMR (400 MHz, methanol-d4) δ = 7.80 (s, 1H), 7.77-7.72 (m, 3H), 7.72-7.66 (m, 3H), 7.25 (d, J = 1.3 Hz, 1H), 6.92 (d, J = 2.2 Hz, 1H), 6.80 (dd, J = 2.6, 8.8 Hz, 1H), 5.27-5.17 (m, 2H), 4.63 (d, J = 12.8 Hz, 1H) z,1H),4.45(d,J=12.8Hz,1H),4.08(quin,J=7.1Hz,1H),3.87-3.78(m,1H),3.74-3.58(m,2 H),3.46(br.s.,1H),3.42(s,3H),3.03-2.95(m,6H),2.67-2.55(m,1H),2.39-2.24(m,1H). ESI[M+H]=479.2

[0445] Plan 25

[0446]

[0447] Chemical experimental methods:

[0448]

[0449] Methyl 3-aminopicolinate, S66. To a suspension of 3-aminopicolinic acid (100 g, 724 mmol, 1.0 eq.) in anhydrous MeOH (1.5 L) was added dropwise concentrated HSO (460 g, 4.69 mol, 6.48 eq.) at 15°C, and the mixture was heated to 80°C and stirred for 5 days. The reaction mixture was concentrated to approximately 500 mL and diluted with cold water (2 L). The resulting mixture was adjusted to approximately 9 with solid NaCO at 0°C and extracted with DCM / MeOH (10:1, 1.0 L x 5). The combined organic layers were dried over NaSO, filtered, and concentrated to yield methyl 3-aminopicolinate (60 g, 394.35 mmol, 54.47% yield, 100% purity) as a brown solid. 1 H NMR (400MHz, CDCl3) δ = 8.08-8.06 (m, 1H), 7.24-7.20 (m, 1H), 7.06-7.01 (m, 1H), 5.75 (br.s, 1H), 3.97 (s, 3H). ESI[M+H]=153.1

[0450]

[0451] 3-Amino-6-bromopicolinic acid methyl ester, S67. To a solution of 3-aminopicolinic acid methyl ester (50 g, 328 mmol, 1.0 eq.) in MeCN (600 mL) was added portionwise NBS (60 g, 337 mmol, 1.0 eq.) at 0° C., and the reaction mixture was stirred at 15° C. for 16 hr. The precipitate was collected by filtration to give 3-amino-6-bromo-pyridine-2-carboxylic acid methyl ester (48 g, 187 mmol, 56.9% yield, 90% purity) as a light yellow solid, which was used directly without further purification. 1 HNMR (400MHz, CDCl3) δ = 7.35 (d, J = 8.4Hz, 1H), 6.96 (d, J = 8.4Hz, 1H), 5.83 (br.s, 1H), 3.96 (s, 3H). ESI[M+H]=231.0 / 233.0

[0452]

[0453] (3-Amino-6-bromopyridin-2-yl)methanol, S68. To a solution of methyl 3-amino-6-bromopicolinate (16 g, 69 mmol, 1.0 eq.) in THF (500 mL) and MeOH (50 mL) was added portionwise NaBH4 (10.5 g, 277 mmol, 4.0 eq.) at 0°C, and the mixture was stirred at 15°C for 16 hr. The mixture was diluted with EtOAc (2 L) and washed with 10% aqueous NH4Cl (300 mL). The aqueous layer was separated and extracted with EtOAc (200 mL x 2). The combined organic layers were dried over Na2SO4 and concentrated to yield (3-amino-6-bromo-2-pyridinyl)methanol (13 g, 57.6 mmol, 83.2% yield, 90% purity) as a light yellow solid. 1 H NMR (400MHz, DMSO-d6) δ7.18 (d, J = 8.4Hz, 1H), 6.96 (d, J = 8.4Hz, 1H), 5.36 (br.s, 1H), 5.19 (t, J = 5.6Hz, 1H), 4.43 (d, J = 5.6Hz, 2H). ESI[M+H]=203.0 / 205.0

[0454]

[0455] Methyl 1-((3-amino-6-bromopyridin-2-yl)methyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate, S69. To a solution of (3-amino-6-bromopyridin-2-yl)methanol (10 g, 49 mmol, 1.0 eq.) and EtN (15 g, 148.2 mmol, 3 eq.) in anhydrous DCM (1 L) was added MsCl (6.50 g, 56.7 mmol, 1.15 eq.) dropwise at -70°C under N2 atmosphere. The mixture was warmed to 15°C and stirred for 30 min. The mixture was then cooled to 0°C and methyl 4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (8.5 g, 37 mmol, 0.76 eq.), Bu4NOH (25% in H2O, 5.0 g, 4.8 mmol, 0.1 eq.), and aqueous NaOH (25% in H2O, 30 g, 187 mmol, 3.8 eq.) were added sequentially. The reaction mixture was warmed to 15°C and stirred for an additional 16 hr. The mixture was washed with brine (1 L), dried over Na2SO4, and then concentrated to yield methyl 1-((3-amino-6-bromopyridin-2-yl)methyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (13 g, crude) as a brown solid, which was used directly. ESI [M+H] = 411.0 / 413.0

[0456]

[0457] 4-(2-Bromo-6-oxo-6,11-dihydro-5H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazon-8-yl)benzonitrile, S70. To a suspension of methyl 1-[(3-amino-6-bromo-2-pyridyl)methyl]-4-(4-cyanophenyl)pyrrole-2-carboxylate (22 g, 24 mmol, 1.0 eq.) in anhydrous toluene (2.0 L) was added dropwise AlMe (2 M in toluene, 65 mL, 5.4 eq.) at 0°C under N2 atmosphere. The reaction mixture was warmed to 15°C and stirred for 16 hr. Cold saturated aqueous NH4Cl solution (500 mL) and water (500 mL) were added. The resulting precipitate was collected by filtration (very little product dissolved in the toluene layer). The crude product was washed with THF (1 L) and the solid was collected by filtration to give 4-(2-bromo-6-oxo-6,11-dihydro-5H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazopentan-8-yl)benzonitrile (12 g, 27.5 mmol, 57% yield, 86.8% purity) as a light yellow solid which was used directly. 1 H NMR (400MHz, DMSO-d6) δ = 10.43 (br.s, 1H), 7.91 (br.s, 1H), 7.77 (br.s, 4H), 7.67-7.65 (m, 1H), 7.56-7.54 (m, 1H), 7.36 (br.s, 1H), 5.37 (s, 2H). ESI[M+H]=379.0.

[0458]

[0459] 4-(2-Bromo-6-chloro-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine-8-yl)benzonitrile, S71. A suspension of 4-(2-bromo-6-oxo-6,11-dihydro-5H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine-8-yl)benzonitrile (2.0 g, 4.54 mmol, 1.0 eq.) in POCl (100 mL) was heated to 100° C. and stirred for 3.5 hr. The reaction mixture was concentrated to give 4-(2-bromo-6-chloro-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine-8-yl)benzonitrile (2.0 g, crude) as a dark yellow solid which was used directly.

[0460]

[0461]

[0149] 4-(7-Bromo-9H-pyrido[3,2-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, S72. A mixture of 4-(2-bromo-6-chloro-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile (1.8 g, 4.5 mmol, 1.0 eq.) and formic hydrazide (1.8 g, 30 mmol, 6.6 eq.) in anhydrous dioxane (40 mL) and anhydrous THF (10 mL) was stirred at 120 °C in a sealed tube for 16 hr. The mixture was concentrated and the residual solid was washed with 1 M HCl (20 mL) and EtOAc (20 mL), filtered and dried in vacuo to give 4-(7-bromo-9H-pyrido[3,2-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopyridine-12-yl)benzonitrile (1.33 g, crude) as a yellow solid which was used directly. ESI [M+H] = 403.0 / 405.0

[0462]

[0463] 4-(7-(4-(Dimethylamino)piperidin-1-yl)-9H-pyrido[3,2-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 293. Synthesized using general procedure A substituting 4-(7-bromo-9H-pyrido[3,2-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and N,N-dimethylpiperidin-4-amine for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 9.07 (br.s., 1H), 7.87 (d, J = 9.0Hz, 1H), 7.77-7.71 (m ,2H),7.70-7.64(m,3H),7.27(br.s.,1H),7.02(d,J=9.0Hz,1H),5.24(s,2H ), 4.72 (d, J = 13.7Hz, 2H), 3.51 (ddd, J = 3.9, 8.3, 11.9Hz, 1H), 3.00 (t, J = 12. 2Hz, 2H), 2.89 (s, 6H), 2.18 (d, J = 11.2Hz, 2H), 1.70 (dq, J = 4.0, 12.1Hz, 2H). ESI[M+H]=451.2

[0464] Plan 26

[0465]

[0466] Chemical experimental methods:

[0467]

[0468]

[0266] 4-(2-Bromo-6-((2,2-dimethoxyethyl)amino)-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile, S73. A mixture of 4-(2-bromo-6-chloro-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile (2.1 g, crude) and 2,2-dimethoxyethylamine (12 g, 100 mmol, 19.07 eq.) in anhydrous dioxane (100 mL) was heated to 100° C. for 16 hr and then concentrated. The residue was dissolved in EtOAc (200 mL) and washed with saturated NaCO (50 mL), 1 M HCl (50 mL), and brine (50 mL). The organic layer was dried and concentrated to give 4-(2-bromo-6-((2,2-dimethoxyethyl)amino)-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile (2.5 g, crude) as a brown solid which was used directly. ESI [M+H] = 466.0 / 468.0

[0469]

[0470]

[0147] 4-(7-Bromo-9H-imidazo[2,1-c]pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, S74. A suspension of 4-(2-bromo-6-((2,2-dimethoxyethyl)amino)-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile (700 mg, 1.50 mmol, 1.0 eq.) in dioxane (25 mL) and 1 M HCl (25 mL) was heated to 60 °C for 10 hr and then concentrated. The residual solid was washed with THF (30 mL) to give 4-(7-bromo-9H-imidazo[2,1-c]pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazopentan-12-yl)benzonitrile (450 mg, 1.04 mmol, 69.36% yield, 93% purity) as a white solid which was used directly without further purification.

[0471]

[0472] (R)-4-(7-(3-(Dimethylamino)pyrrolidin-1-yl)-9H-imidazo[2,1-c]pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 312. Synthesized using general procedure A, substituting 4-(7-bromo-9H-imidazo[2,1-c]pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (R)-N,N-dimethylpyrrolidin-3-amine for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 8.06-7.99 (m, 1H), 7.93 (d, J = 9.0Hz, 1H), 7.84 (s, 1H),7.78-7.76(m,3H),7.73-7.68(m,2H),7.36(s,1H),6.77(d,J=9.0Hz ,1H),5.37(s,2H),4.16-4.04(m,2H),3.91-3.83(m,1H),3.82-3.73(m,1 H),3.65-3.55(m,1H),3.00(s,6H),2.68-2.56(m,1H),2.41-2.29(m,1H). ESI[M+H]=436.1

[0473] Plan 27

[0474]

[0475] Chemical experimental methods:

[0476]

[0477] (E)-2-(2-chloro-5-nitropyridin-4-yl)-N,N-dimethylethanamine, S76. To a solution of 2-chloro-4-methyl-5-nitropyridine (40 g, 231 mmol, 1.0 eq.) in DMF (200 mL) was added DMF-DMA (55 g, 463 mmol, 2.0 eq.) at 20° C., and the mixture was stirred at 90° C. for 5 hr. The mixture was then poured into cold water (300 mL) and the precipitate was collected by filtration to give crude (E)-2-(2-chloro-5-nitro-4-pyridinyl)-N,N-dimethylethanamine (44.3 g, crude) as a red solid, which was not further purified.

[0478]

[0479] 2-Chloro-5-nitroisonicotinaldehyde, S77. To a solution of (E)-2-(2-chloro-5-nitro-4-pyridyl)-N,N-dimethyl-ethanamine (44.3 g, 194.6 mmol, 1.0 eq.) in THF (250 mL) and H₂O (250 mL) was added portionwise NaIO₄ (145 g, 681 mmol, 3.5 eq.) at 20°C, and the reaction mixture was stirred at 20°C for 16 hr. The mixture was then filtered and the filter cake was washed with EtOAc (100 mL x 2). The filtrate was extracted with EtOAc (500 mL x 2). The combined organic layers were washed with brine (300 mL), dried over Na₂SO₄, and concentrated to yield 2-chloro-5-nitroisonicotinaldehyde (37.2 g, 92% yield, 90% purity) as a dark brown oil. 1H NMR (400 MHz, chloroform-d) δ=10.49 (br.s., 1H), 9.23 (br.s., 1H), 7.74 (br.s., 1H).

[0480]

[0481] (2-Chloro-5-nitropyridin-4-yl)methanol, S78. To a solution of 2-chloro-5-nitroisonicotinaldehyde (40 g, 214 mmol, 1.0 eq.) in MeOH (300 mL) was added portionwise NaBH4 (12.1 g, 321.6 mmol, 1.5 eq.) at 0°C. The mixture was stirred at 0°C for 3 hr and quenched by saturated NH4Cl (200 mL). After removing MeOH, the aqueous layer was extracted with EtOAc (200 mL*2). The organic layer was washed with brine (200 mL), dried over MgSO4, and concentrated. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate 20:1 to 10:1) to give (2-chloro-5-nitropyridin-4-yl)methanol (19.5 g, 103.4 mmol, 48.2% yield) as a yellow solid. 1H NMR (400 MHz, CHLOROFORM-d) δ=9.12 (br.s., 1H), 7.92 (br.s., 1H), 5.16-5.14 (m, J=2.2 Hz, 2H).

[0482]

[0483] (5-amino-2-chloropyridin-4-yl)methanol, S79. To a solution of (2-chloro-5-nitropyridin-4-yl)methanol (16 g, 84.8 mmol, 1.0 eq.) in EtOH (160 mL) and THF (160 mL) was added Fe (23.6 g, 424 mmol, 5.0 eq.) and saturated NH4Cl (80 mL). The mixture was stirred at 90 ° C for 1 hr and filtered. The filtrate was concentrated and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 2: 1, Rf = 0.12) to give (5-amino-2-chloropyridin-4-yl)methanol (12.9 g, 74.83 mmol, 88.19% yield, 92% purity) as a yellow solid. ESI [M+H] = 158.8

[0484]

[0485] Methyl 1-((5-amino-2-chloropyridin-4-yl)methyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate, S80. To a solution of (5-amino-2-chloropyridin-4-yl)methanol (4.0 g, 25.2 mmol, 1.0 eq.) and TEA (7.6 g, 75.6 mmol, 3.0 eq.) in DCM (320 mL) was added MsCl (4.3 g, 37.8 mmol, 1.5 eq.) at -70°C. The mixture was slowly warmed to 20°C and stirred for 3 hr. Methyl 4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (4.5 g, 20.1 mmol, 0.8 eq.) was then added at 0°C, followed by aqueous NaOH (25%, 6.00 eq.) and tetrabutylammonium hydroxide (2.62 g, 2.52 mmol, 0.1 eq.). After addition, the mixture was stirred at 15°C for 16 hours. The resulting mixture was then diluted with water (500 mL) and extracted with DCM (100 mL x 2). The combined organic layers were dried over anhydrous MgSO₄, filtered, and concentrated to afford methyl 1-((5-amino-2-chloropyridin-4-yl)methyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (2.2 g, 4.4 mmol, 17.6% yield, 74% purity) as a yellow solid. 1H NMR (400 MHz, chloroform-d) δ = 7.86 (s, 1H), 7.64-7.62 (m, 2H), 7.58-7.56 (m, 2H), 7.34-7.33 (m, 1H), 7.21 (br.s., 1H), 6.71 (br.s., 1H), 5.45 (br.s., 2H), 3.86-3.85 (m, 3H). ESI [M+H] = 367.0

[0486]

[0487] 4-(3-Chloro-10-oxo-10,11-dihydro-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazon-8-yl)benzonitrile, S81. To a solution of methyl 1-((5-amino-2-chloropyridin-4-yl)methyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (2.3 g, 6.2 mmol, 1.0 eq.) in toluene (25 mL) was added AlMe (2 M, 15.6 mL, 5.0 eq.), and the mixture was stirred at 20° C. for 16 hr. The mixture was poured into cold water (100 mL) and extracted with EtOAc (200 mL*2). The organic layer was washed with brine (100 mL), dried over MgSO, and concentrated. The resulting solid was washed with EtOH (20 mL). After filtration, the filter cake was collected to yield 4-(3-chloro-10-oxo-10,11-dihydro-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile (1.82 g, 4.73 mmol, 75.44% yield, 87% purity) as a yellow solid. ESI [M+H] = 334.9

[0488]

[0489] 4-(3-Chloro-10-thione-10,11-dihydro-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile, S82. To a solution of 4-(3-chloro-10-oxo-10,11-dihydro-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile (1.8 g, 5.4 mmol, 1.0 eq.) in THF (20 mL) was added Lawesson's reagent (2.6 g, 6.5 mmol, 1.2 eq.) and the mixture was stirred at 70° C. for 1 hr. The mixture was concentrated and the residue was washed with MeOH (20 mL). After filtration, the filter cake was collected and concentrated to give crude 4-(3-chloro-10-thione-10,11-dihydro-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazopentan-8-yl)benzonitrile (1.39 g, crude) as a yellow solid.

[0490]

[0491] 4-(3-Chloro-10-(methylthio)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile, S83. To a suspension of 4-(3-chloro-10-thione-10,11-dihydro-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile (1.4 g, 3.9 mmol, 1.0 eq.) and K2CO3 (3.8 g, 27.7 mmol, 7.0 eq.) in DMF (20 mL) was added MeI (1.1 g, 7.9 mmol, 2.0 eq.) at 0° C. The mixture was stirred at 20° C. for 1 hr and filtered. The filtrate was concentrated to give 4-(3-chloro-10-(methylthio)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazopentan-8-yl)benzonitrile (1.3 g, crude) as a yellow solid.

[0492]

[0493] 4-(7-Chloro-9H-pyrido[3,4-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, S84. A mixture of 4-(3-chloro-10-(methylthio)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile (535 mg, 8.9 mmol, 2.5 eq.) and formic hydrazide (3.0 g) in n-BuOH (20 mL) was stirred at 130° C. for 16 hr and concentrated. The residue was washed with 1N HCl (20 mL) and EtOH (10 mL). After filtration, the filter cake was collected to yield 4-(7-chloro-9H-pyrido[3,4-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopyridine-12-yl)benzonitrile (600 mg, 33.3% yield, 71% purity) as a yellow solid. ESI [M+H] = 358.9

[0494]

[0495]

[0147] 4-(7-(4-(Dimethylamino)piperidin-1-yl)-9H-pyrido[3,4-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 321. Synthesized using general procedure A, substituting 4-(7-chloro-9H-pyrido[3,4-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and N,N-dimethylpiperidin-4-amine for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 9.15 (br.s., 1H), 8.42 (s, 1H), 7.76-7.58 (m, 5H), 7.26 (br.s., 1H), 7.13 (s, 1H), 5.22 (s, 2H), 4.66 (d, J = 13.7Hz, 2H), 3.51 (ddd, J=3.9, 8.3, 11.9Hz, 1H), 3.01 (t, J=12.5Hz, 2H), 2.92-2.80 (m, 6H), 2.16 (d, J=11.5Hz, 2H), 1.70 (dq, J=4.0, 12.1Hz, 2H). ESI[M+H]=451.2

[0496] Plan 28

[0497]

[0498] Chemical experimental methods:

[0499]

[0500] 4-(3,10-Dichloro-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazopentan-8-yl)benzonitrile, S85. A solution of 4-(3-chloro-10-oxo-10,11-dihydro-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazopentan-8-yl)benzonitrile (500 mg, 1.49 mmol, 1.0 eq.) in POCl (30 mL) was stirred at 90° C. for 1 hr and concentrated to give 4-(3,10-dichloro-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazopentan-8-yl)benzonitrile (550 mg, crude, HCl) as a dark brown oil.

[0501]

[0502]

[0145] 4-(3-Chloro-10-((2,2-dimethoxyethyl)amino)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile, S86. A solution of 4-(3,10-dichloro-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile (550 mg, 1.41 mmol, 1.0 eq., HCl) in THF (10 mL) and dioxane (10 mL) was adjusted to pH = 9 with TEA, followed by the addition of 2,2-dimethoxyethylamine (1.48 g, 14.1 mmol, 1.53 mL, 10.0 eq.) and the mixture was stirred at 120 °C in a sealed tube for 16 hr. After the reaction was complete, the mixture was poured into 0.5M HCl (50 mL) and extracted with hot EtOAc / THF=1 / 1 (50 mL*3). The organic layer was washed with brine (50 mL), dried over MgSO4 and concentrated. The residue was purified by column chromatography (petroleum ether:ethyl acetate=1:1, Rf=0.53) to produce 4-(3-chloro-10-((2,2-dimethoxyethyl)amino)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine-8-yl)benzonitrile (130 mg, 150 umol, 10.6% yield, 48.8% purity) as a yellow solid. ESI[M+H]=422.1

[0503]

[0504] 4-(7-Chloro-9H-imidazo[2,1-c]pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, S87. A solution of 4-(3-chloro-10-((2,2-dimethoxyethyl)amino)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile (130 mg, 150 umol, 1.0 eq.) in dioxane (6.0 mL) and 1 M HCl (6.0 mL) was stirred at 80° C. for 16 hr. After completion of the reaction, the mixture was concentrated. The residual solid was washed with THF (5 mL) and filtered. The filter cake was collected to give the crude product 4-(7-chloro-9H-imidazo[2,1-c]pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile (70 mg, crude, HCl) as a black solid, which was not further purified. ESI [M+H] = 358.1

[0505]

[0506]

[0147] 4-(7-(2,6-diazaspiro[3.4]octan-6-yl)-9H-imidazo[2,1-c]pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile, 323. Synthesized using general procedure A, substituting 4-(7-chloro-9H-imidazo[2,1-c]pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile and tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 8.40 (s, 1H), 8.08 (m, 1H), 7.80-7.68 (d, J = 9.9Hz, 6H), 7.34 (d, J = 1.8Hz, 1H), 7.33 (d, J = 1.1H z,1H),5.11(s,2H),4.00-3.92(m,2H),3.91-3.83(m,2H),3.73(s,2H),3.54(t,J=6.9Hz,2H),2.33(t,J=6.9Hz,2H). ESI[M+H]=434.1

[0507] Plan 29

[0508]

[0509] Chemical experimental methods:

[0510]

[0511] (4,5-difluoro-2-nitrophenyl)methanol, S90. To a solution of 4,5-difluoro-2-nitrobenzoic acid (8.0 g, 39.39 mmol, 1.0 eq.) in THF (80 mL) was added dropwise a solution of BH3-Me2S (10 M, 11.82 mL, 3.0 eq.) at 0°C over a period of 30 min under N2. The reaction mixture was heated to 70°C over a period of 30 min and stirred at 70°C for 3 hr. The reaction was slowly quenched by ice water and then extracted with ethyl acetate (50 mL*3). The combined organic phases were washed with brine (10 mL*2), dried over Na2SO4, filtered and concentrated to give (4,5-difluoro-2-nitrophenyl)methanol (7.2 g, crude) as a yellow solid. TLC: R f =0.6 (petroleum ether / ethyl acetate=3 / 1).

[0512]

[0513] tert-Butyl 4-(2-fluoro-5-(hydroxymethyl)-4-nitrophenyl)piperazine-1-carboxylate, S91. A mixture of (4,5-difluoro-2-nitrophenyl)methanol (7.6 g, 40.3 mmol, 1.00 eq.) and tert-butyl piperazine-1-carboxylate (9.0 g, 48.4 mmol, 1.2 eq.) in DMSO (80 mL) was heated to 90° C. for 3 hr and poured into ice water (100 mL). The aqueous phase was extracted with ethyl acetate (200 mL*3). The combined organic phases were washed with brine (20 mL*2), dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give tert-butyl 4-(2-fluoro-5-(hydroxymethyl)-4-nitrophenyl)piperazine-1-carboxylate (11.26 g, crude) as a yellow solid. TLC: R f =0.35 (petroleum ether / ethyl acetate=3 / 1). 1H NMR (400 MHz, chloroform-d) δ=7.85 (d, J=13.2 Hz, 1H), 7.09 (d, J=8.4 Hz, 1H), 4.89 (br. s., 2H), 3.61-3.49 (m, 4H), 3.31-3.14 (m, 4H), 1.42 (s, 9H).

[0514]

[0515] tert-Butyl 4-(5-((4-(4-cyanophenyl)-2-(methoxycarbonyl)-1H-pyrrol-1-yl)methyl)-2-fluoro-4-nitrophenyl)piperazine-1-carboxylate, S92. To a mixture of tert-butyl 4-(2-fluoro-5-(hydroxymethyl)-4-nitrophenyl)piperazine-1-carboxylate (863 mg, 2.43 mmol, 1.1 eq.) and methyl 4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (500 mg, 2.21 mmol, 1.0 eq.) in Tol. (10 mL) was added 2-(tributylphosphinilidene)acetonitrile (1.07 g, 4.42 mmol, 2.0 eq.) in one portion at 30° C. under N2. The mixture was stirred at 30° C. for 30 min, then heated to 100° C. and stirred for 11.5 hr. The mixture was concentrated and diluted with ice water (50 mL). The aqueous phase was extracted with ethyl acetate (50 mL * 3). The combined organic phase was washed with brine (10 mL * 2), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by preparative HPLC to give tert-butyl 4-(5-((4-(4-cyanophenyl)-2-(methoxycarbonyl)-1H-pyrrol-1-yl)methyl)-2-fluoro-4-nitrophenyl)piperazine-1-carboxylate (600 mg, 1.0 mmol, 45.76% yield, 95% purity) as a yellow solid. TLC: R f=0.3 (petroleum ether / ethyl acetate = 3 / 1). 1H NMR (400 MHz, chloroform-d) δ = 7.96 (d, J = 13.2 Hz, 1H), 7.69-7.57 (m, 4H), 7.39 (s, 1H), 7.29 (s, 2H), 5.99 (s, 2H), 3.79 (s, 3H), 3.53-3.42 (m, 4H), 3.02 (br. s., 4H), 1.44 (s, 9H). ESI [M+H] = 564.5

[0516]

[0517] tert-Butyl 4-(4-amino-5-((4-(4-cyanophenyl)-2-(methoxycarbonyl)-1H-pyrrol-1-yl)methyl)-2-fluorophenyl)piperazine-1-carboxylate, S93. To a mixture of tert-butyl 4-(5-((4-(4-cyanophenyl)-2-(methoxycarbonyl)-1H-pyrrol-1-yl)methyl)-2-fluoro-4-nitrophenyl)piperazine-1-carboxylate (620 mg, 1.1 mmol, 1.0 eq.) in AcOH (10 mL) was added Zn (719 mg, 11.0 mmol, 10.0 eq.) in one portion at 30° C. under N 2 . The mixture was stirred at 30° C. for 1 hr, filtered, and concentrated. The residue was diluted with ice water (20 mL) and extracted with ethyl acetate (15 mL*3). The combined organic phases were washed with brine (5 mL * 2), dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give tert-butyl 4-(4-amino-5-((4-(4-cyanophenyl)-2-(methoxycarbonyl)-1H-pyrrol-1-yl)methyl)-2-fluorophenyl)piperazine-1-carboxylate (580 mg, crude) as a yellow solid. TLC: R f =0.26 (petroleum ether / ethyl acetate=3 / 1). ESI[M+H]=533.9

[0518]

[0519] 4-(8-Fluoro-11-oxo-7-(piperazin-1-yl)-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-2-yl)benzonitrile, S94. To a solution of tert-butyl 4-(4-amino-5-((4-(4-cyanophenyl)-2-(methoxycarbonyl)-1H-pyrrol-1-yl)methyl)-2-fluorophenyl)piperazine-1-carboxylate (300 mg, 562 umol, 1.0 eq.) in toluene (20 mL) was added dropwise a solution of AlMe3 (2 M, 1.41 mL, 5.0 eq.) at 0°C over a period of 30 min under N2. The reaction mixture was warmed to 30°C and stirred for 3 hr. The reaction was slowly quenched with ice water (30 mL) and extracted with DCM (30 mL*3). The combined organic phases were washed with saturated brine (5 mL*2), dried over anhydrous Na2SO4, filtered and concentrated to yield crude 4-(8-fluoro-11-oxo-7-(piperazin-1-yl)-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)benzonitrile (140 mg, crude) as a yellow solid. TLC: R f =0.46 (petroleum ether / ethyl acetate=3 / 1). ESI[M+H]=402.1

[0520]

[0521]

[0266] Tert-butyl 4-(2-(4-cyanophenyl)-8-fluoro-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazopentan-7-yl)piperazine-1-carboxylate, S95. To a solution of 4-(8-fluoro-11-oxo-7-(piperazin-1-yl)-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazopentan-2-yl)benzonitrile (150 mg, 373 umol, 1.0 eq.) in MeOH (5.0 mL) was added BocO (97 mg, 448 umol, 1.2 eq.) in one portion. The mixture was stirred at 30° C. for 2 hours and concentrated under reduced pressure to give tert-butyl 4-(2-(4-cyanophenyl)-8-fluoro-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-7-yl)piperazine-1-carboxylate (200 mg, crude) as a yellow solid. TLC: R f =0.53 (petroleum ether / ethyl acetate=3 / 1). ESI[M+H]=502.2

[0522]

[0523]

[0266] tert-Butyl 4-(2-(4-cyanophenyl)-8-fluoro-11-thione-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-7-yl)piperazine-1-carboxylate, S96. A solution of tert-butyl 4-(2-(4-cyanophenyl)-8-fluoro-11-oxo-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-7-yl)piperazine-1-carboxylate (200 mg, 398 umol, 1.0 eq.) and Lawesson's reagent (193 mg, 478 umol, 1.2 eq.) in THF (10.0 mL) was stirred at 80 °C for 16 hr and concentrated. The residue was purified by preparative TLC (DCM / methanol = 15 / 1) to give tert-butyl 4-(2-(4-cyanophenyl)-8-fluoro-11-thione-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-7-yl)piperazine-1-carboxylate (80 mg, 123 umol, 31% yield, 80% purity) as a white solid. TLC: R f =0.33 (dichloromethane / methanol=20 / 1).

[0524]

[0525] tert-Butyl 4-(2-(4-cyanophenyl)-8-fluoro-11-(methylthio)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-7-yl)piperazine-1-carboxylate, S97. To a mixture of tert-butyl 4-(2-(4-cyanophenyl)-8-fluoro-11-thione-10,11-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-7-yl)piperazine-1-carboxylate (80 mg, 154 umol, 1.0 eq.) and K2CO3 (42.7 mg, 309 umol, 2.0 eq.) in MeCN (3.0 mL) was added MeI (109 mg, 772 umol, 5.0 eq.). The mixture was stirred at 28° C. for 16 hr and concentrated in vacuo. The residue was purified by preparative TLC (petroleum ether / ethyl acetate = 2 / 1) to give tert-butyl 4-(2-(4-cyanophenyl)-8-fluoro-11-(methylthio)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-7-yl)piperazine-1-carboxylate (75 mg, 112 umol, 73% yield, 80% purity) as an orange oil. TLC: R f =0.23 (petroleum ether / ethyl acetate=3 / 1).

[0526]

[0527] tert-Butyl 4-(12-(4-cyanophenyl)-6-fluoro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)piperazine-1-carboxylate, S98. Formic acid hydrazide (18 mg, 300 umol, 2.0 eq.) and 4-(2-(4-cyanophenyl)-8-fluoro-11-(methylthio)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-7-yl)piperazine were reacted with 4-(2-(4-cyanophenyl)-8-fluoro-11-(methylthio)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-7-yl)piperazine. A mixture of tert-butyl-1-carboxylate (80 mg, 150 umol, 1.0 eq.) in n-BuOH (2.0 mL) was stirred at 130° C. for 16 hr and concentrated under reduced pressure to give tert-butyl-4-(12-(4-cyanophenyl)-6-fluoro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)piperazine-1-carboxylate (50 mg, crude), which was not further purified. TLC: R f =0.05 (petroleum ether / ethyl acetate=3 / 1).

[0528]

[0529] tert-Butyl 4-(12-(4-cyanophenyl)-6-fluoro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)piperazine-1-carboxylate (80 mg, 152 umol, 1.0 eq.) was dissolved in HCl / EtOAc (4 M, 3.0 mL) and the mixture was stirred at 28° C. for 0.5 hr. The mixture was concentrated and the residue was purified by preparative HPLC to yield 4-(6-fluoro-7-(piperazin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile (30 mg, 70 umol, 46.3% yield, 100% purity). 1H NMR (400 MHz, Methanol-d4) δ = 9.07 (s, 1H), 7.77-7.65 (m, 4H), 7.65-7.58 (m, 2H), 7.41 (d, J = 8.8 Hz, 1H), 7.26 (s, 1H), 5.25 (s, 2H), 3.50-3.39 (m, 8H). ESI [M+H] = 426.1

[0530] Plan 30

[0531]

[0532] Chemical Experimental Methods

[0533]

[0534] 3-amino-5-fluoropicolinic acid methyl ester, S100. A mixture of 2-bromo-5-fluoropyridine-3-amine (22 g, 115.18 mmol, 1.00 eq.), Et3N (23.3 g, 230 mmol, 2.0 eq.) and Pd(dppf)Cl2 (4.2 g, 5.7 mmol, 0.05 eq.) in MeOH (1 L) was degassed and stirred at 80 ° C under CO (2 MPa) for 50 hr. The mixture was concentrated and ice water (300 mL) was added. The aqueous phase was extracted with ethyl acetate (200 mL*3). The phase was washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography to produce 3-amino-5-fluoropicolinic acid methyl ester (15 g, 74.3 mmol, 64.5% yield) as a yellow solid. 1H NMR (400 MHz, chloroform-d) δ = 7.92 (d, J = 2.0 Hz, 1H), 6.74 (dd, J = 2.3, 9.8 Hz, 1H), 5.96 (br. s., 2H), 3.98 (s, 3H). ESI [M+H] = 171.1

[0535]

[0536] 3-amino-6-bromo-5-fluoropicolinic acid methyl ester, S101. To a solution of 3-amino-5-fluoropicolinic acid methyl ester (15 g, 88 mmol, 1.0 eq.) in MeCN (500 mL) was added NBS (15.7 g, 88.4 mmol, 1.0 eq.) and the mixture was stirred at 15 ° C for 14 hr. The mixture was concentrated and the residue was partitioned between EtOAc (500 mL) and ice water (100 mL). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography to produce 3-amino-6-bromo-5-fluoropicolinic acid methyl ester (18 g, 62 mmol, 70.27% yield) as a white solid. 1H NMR (400 MHz, chloroform-d) δ = 6.79 (d, J = 8.4 Hz, 1H), 5.98 (br. s., 2H), 3.95 (s, 3H). ESI [M+H] = 249.2 / 251.2

[0537]

[0538] (3-Amino-6-bromo-5-fluoropyridin-2-yl)methanol, S102. To a solution of methyl 3-amino-6-bromo-5-fluoropicolinate (13.8 g, 55.5 mmol, 1.0 eq.) in THF (200 mL) was added aqueous LiOH (11.6 g, 277 mmol, 5.00 eq., 1 M in H O) and the mixture was stirred at 15 ° C for 2 hr. The solution was adjusted to pH = 2 with 4M HCl and then concentrated. The residue was partitioned between EtOAc (300 mL) and ice water (100 mL). The organic phase was washed with brine (20 mL), dried over anhydrous Na SO, filtered and concentrated to give 3-amino-6-bromo-5-fluoropicolinic acid (13 g, crude) as a yellow solid, which was used in the next step without further purification.

[0539] To a solution of 3-amino-6-bromo-5-fluoropicolinic acid (11.0 g, 46.8 mmol, 1.0 eq.) in THF (20 mL) was added BH3.THF (1 M, 300 mL, 6.4 eq.) and the mixture was heated to 70 ° C and maintained for 16 hr. The mixture was cooled to 0 ° C, slowly quenched by MeOH (400 mL) and then concentrated in vacuo. MeOH (30 mL) was added to the residue and stirred for 1 hr. The precipitate was collected by filtration to produce (3-amino-6-bromo-5-fluoropyridin-2-yl)methanol (8.0 g, 36.19 mmol, 77.31% yield) as a white solid. ESI [M + H] = 221.2 / 223.2

[0540]

[0541] (3-Amino-6-bromo-5-fluoropyridin-2-yl)methyl methanesulfonate, S103. To a solution of (3-amino-6-bromo-5-fluoropyridin-2-yl)methanol (6 g, 27.12 mmol, 1.0 eq.) and TEA (5.49 g, 54.24 mmol, 2.0 eq.) in DCM (200 mL) was added dropwise methanesulfonyl chloride (3.26 g, 28.5 mmol, 1.05 eq.) at -70 °C under N2. After the addition, the mixture was warmed to 15 °C and stirred for 30 min. The solution was used directly in the next step.

[0542]

[0543] Methyl 1-((3-amino-6-bromo-5-fluoropyridin-2-yl)methyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate, S104. To the above solution were added methyl 4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (4.9 g, 21.8 mmol, 0.80 eq.), Bu4NOH (2.6 g, 2.7 mmol, 0.1 eq.), and NaOH (10.4 g, 163.7 mmol, 6.0 eq. 25%, w %) at 0°C. The mixture was stirred at 15°C for 3.5 hr and then diluted with ice water (100 mL). The organic layer was washed with brine (100 mL*2), dried over anhydrous MgSO4, filtered, and concentrated in vacuo. The residue was purified by preparative HPLC to give methyl 1-((3-amino-6-bromo-5-fluoropyridin-2-yl)methyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (3 g, 4.9 mmol) as a yellow solid. 1H NMR (400 MHz, CHLOROFORM-d) δ = 7.56-7.52 (m, 2H), 7.49 (s, 1H), 7.47-7.45 (m, 1H), 7.19 (s, 2H), 6.67 (d, J = 8.8 Hz, 1H), 5.52 (s, 2H), 3.83 (s, 3H). ESI [M+H] = 429.1 / 431.1

[0544]

[0545] 4-(2-Bromo-3-fluoro-6-oxo-6,11-dihydro-5H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazin-8-yl)benzonitrile, S105. To a suspension of methyl 1-((3-amino-6-bromo-5-fluoropyridin-2-yl)methyl)-4-(4-cyanophenyl)-1H-pyrrole-2-carboxylate (2.80 g, 6.52 mmol, 1.0 eq.) in toluene (50 mL) was added MeAl (2 M in toluene, 16.30 mL, 5.0 eq.) at 0°C and the mixture was stirred at 15°C for 12 hr. The mixture was poured into ice water (100 mL) and extracted with hot EtOAc / THF (1:1, 100 mL*4). The organic layer was dried over anhydrous NaSO, filtered, and concentrated. THF (20 mL) was added to the residue and stirred at 0°C for 1 hr. The precipitate was collected by filtration to give 4-(2-bromo-3-fluoro-6-oxo-6,11-dihydro-5H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile (1.0 g, 2.11 mmol, 32.44% yield) as a white solid. ESI [M+H] = 397.1 / 399.1

[0546]

[0547] 4-(2-Bromo-6-chloro-3-fluoro-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazopentan-8-yl)benzonitrile, S106. A solution of 4-(2-bromo-3-fluoro-6-oxo-6,11-dihydro-5H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazopentan-8-yl)benzonitrile (550 mg, 1.38 mmol, 1.0 eq.) in POCl (30 mL) was stirred at 90° C. for 2 hr. The solution was concentrated under reduced pressure to give crude 4-(2-bromo-6-chloro-3-fluoro-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazopentan-8-yl)benzonitrile (575.00 mg, crude) as a dark brown oil, which was used in the next step without further purification.

[0548]

[0549] 4-(7-Bromo-6-fluoro-9H-pyrido[3,2-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, S107. To a solution of 4-(2-bromo-6-chloro-3-fluoro-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepin-8-yl)benzonitrile (575 mg, 1.38 mmol, 1.0 eq.) in dioxane (50 mL) was added TEA to adjust the pH to 7. Formic hydrazide (1.7 g, 28.9 mmol, 21.0 eq.) was then added and the mixture was heated to 120° C. in a sealed tube for 10 hr. The mixture was concentrated to dryness and the residue was dissolved in hot EtOAc / THF (1:1, 200 mL). The solution was washed with 1M glacial HCl (10 mL), saturated aqueous NaHCO₃ (10 mL), and brine (10 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered, and concentrated in vacuo. EtOH (5 mL) was added to the residue and stirred for 1 hr. The precipitate was collected by filtration to yield 4-(7-bromo-6-fluoro-9H-pyrido[3,2-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopyridine-12-yl)benzonitrile (450 mg, crude) as a white solid. 1H NMR (400 MHz, DMSO-d₆) δ = 9.21 (s, 1H), 8.45 (d, J = 8.4 Hz, 1H), 7.94 (br. s., 1H), 7.76 (s, 4H), 7.31 (s, 1H), 5.43 (br. s., 2H). ESI[M+H]=421.2 / 423.2

[0550]

[0551] (S)-4-(7-(3-(dimethylamino)pyrrolidin-1-yl)-6-fluoro-9H-pyrido[3,2-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 328. General Procedure A was used with 4-(7-bromo-6-fluoro-9H-pyrido[3,2-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile. The product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile with (S)-N,N-dimethylpyrrolidin-3-amine instead of tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 9.04 (br.s., 1H), 7.87 (d, J = 12.3Hz, 1H), 7.76-7.60 (m, 5H), 7.26 (br.s., 1H), 5.23 (s, 2H), 4.24-4.12 (m, 1 H), 4.09-3.97 (m, 2H), 3.96-3.87 (m, 1H), 3.78 (d, J = 9.3Hz, 1H), 3.01 (s, 6H), 2.56 (d, J = 6.2Hz, 1H), 2.29 (dd, J = 8.6, 12.6Hz, 1H). ESI[M+H]=455.2

[0552] Plan 31

[0553]

[0554]

[0555] Chemical experimental methods:

[0556]

[0557] To a mixture of 1-tert-butyl 2-methyl 4-bromo-1H-pyrrole-1,2-dicarboxylate (59.8 g, 196.6 mmol, 1.0 eq.) and (4-fluorophenyl)boronic acid (41 g, 294 mmol, 1.5 eq.) in dioxane (1.8 L) and H2O (180 mL) was added Boc2O (64.3 g, 294.9 mmol, 1.50 eq.), Na2CO3 (41.68 g, 393.24 mmol, 2.00 eq.), and Pd(dppf)Cl2 (7.1 g, 9.8 mmol, 0.05 eq.). The mixture was stirred at 100°C for 12 hr and then concentrated. The residue was partitioned between ethyl acetate / THF (700 mL / 100 mL) and water (500 mL). The organic layer was washed with brine (400 mL), dried over anhydrous MgSO 4 , filtered and concentrated to provide 1-tert-butyl 2-methyl 4-(4-fluorophenyl)-1H-pyrrole-1,2-dicarboxylate (75 g, crude, black solid). ESI [M+H] = 320.1

[0558]

[0559] Methyl 4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate, S109. A solution of 1-tert-butyl 2-methyl 4-(4-fluorophenyl)-1H-pyrrole-1,2-dicarboxylate (75 g, 234 mmol, 1.0 eq.) in TFA (500 mL) was stirred at 50 °C for 1 hr and then concentrated in vacuo. MeOH (250 mL) was added to the residue and stirred for 1 hr. The precipitate was collected by filtration to yield methyl 4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate (25 g, 112 mmol, 47.8% yield, 98.6% purity) as a gray solid. 1HNMR (400 MHz, methanol-d4) δ = 7.55 (dd, J = 8.8, 5.6 Hz, 2H), 7.30 (d, J = 1.2 Hz, 1H), 7.15 (d, J = 1.6 Hz, 1H), 7.06 (t, J = 8.8 Hz, 2H), 3.86 (s, 3H). ESI [M+H] = 220.1

[0560]

[0561] Methyl 1-(5-bromo-2-nitrobenzyl)-4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate, S110. To a solution of (5-bromo-2-nitrophenyl)methanol (30 g, 129 mmol, 1.0 eq.) and TEA (19 g, 193 mmol, 1.50 eq.) in DCM (600 mL) was added MsCl (16 g, 142 mmol, 1.1 eq.) at 0° C., and the mixture was stirred at 20° C. for 30 min. Methyl 4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate (24 g, 110 mmol, 0.95 eq.) was then added at 0° C., followed by NaOH (25% aqueous solution, 5.00 eq.) and tetrabutylammonium hydroxide (12.0 g, 11.6 mmol, 0.10 eq.). The mixture was stirred at 20°C for 16 hours and then poured into cold water (800 mL) and extracted with DCM (300 mL*3). The organic layer was dried over MgSO4 and concentrated in vacuo. EtOH / MeOH (220 mL, 10 / 1) was added to the residue and stirred for 1 hour. The precipitate was collected by filtration to provide methyl 1-(5-bromo-2-nitrobenzyl)-4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate (48.6 g, 107.45 mmol, 92.57% yield, 95.78% purity) as a yellow solid. 1H NMR (400 MHz, chloroform-d) δ = 8.05 (d, J = 8.8 Hz, 1H), 7.53 (d, J = 5.6 Hz, 1H), 7.52-7.50 (m, 2H), 7.33 (d, J = 2.0 Hz, 1H), 7.19 (d, J = 2.0 Hz, 1H), 7.10 (t, J = 8.4 Hz, 2H), 6.77 (s, 1H), 5.95 (s, 2H), 3.78 (s, 3H). ESI [M+H] = 435.0

[0562]

[0563] 1-(2-amino-5-bromobenzyl)-4-(4-fluorophenyl)-1H-pyrrole-2-carboxylic acid methyl ester, S111. To a solution of 1-(5-bromo-2-nitrobenzyl)-4-(4-fluorophenyl)-1H-pyrrole-2-carboxylic acid methyl ester (47.60 g, 109.87 mmol, 1.0 eq.) in THF (480 mL) and EtOH (480 mL) was added Fe (30.68 g, 549.35 mmol, 5.0 eq.) and saturated aqueous NH4Cl solution (240 mL). The mixture was stirred at 90 ° C for 30 min and concentrated in vacuo. The residue was dissolved in hot THF (2 L). After filtration, the filtrate was concentrated to provide 1-(2-amino-5-bromobenzyl)-4-(4-fluorophenyl)-1H-pyrrole-2-carboxylic acid methyl ester (66 g, crude) as a yellow solid. TLC:R f =0.25 (petroleum ether:ethyl acetate=5:1).

[0564]

[0565] 7-Bromo-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one, S112. To a suspension of methyl 1-(2-amino-5-bromobenzyl)-4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate (65 g, 101.55 mmol, 1.0 eq.) in toluene (650 mL) was added AlMe (2 M, 253.88 mL, 5.0 eq.) and the mixture was stirred at 20° C. for 16 hr. The mixture was quenched with 1 M glacial HCl (2 L) and extracted with ethyl acetate / THF (2 / 1, 1000 mL*3). The organic layer was washed with brine (1 L), dried over MgSO, and concentrated in vacuo. To the residue was added EtOH / MeOH (220 mL) and filtered. The filter cake was collected and dried to provide 7-bromo-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one (28 g, 67.89 mmol, 66.85% yield, 90% purity) as a yellow solid. ESI [M+H] = 370.9 / 372.9

[0566]

[0567] 7-Bromo-11-chloro-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine, S113. A solution of 7-bromo-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one (2.00 g, 5.39 mmol, 1.0 eq.) in POCl (50 mL) was stirred at 90° C. for 1 hr and then concentrated to give 7-bromo-11-chloro-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine (2.3 g, crude) as a dark brown oil.

[0568]

[0569] 7-Bromo-12-(4-fluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine, S114. A solution of 7-bromo-11-chloro-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine (2.30 g, 5.90 mmol, 1.0 eq.) in THF (20 mL) and dioxane (20.00 mL) was adjusted to pH = 9 with TEA. Formic hydrazide (5.45 g, 90 mmol, 20.0 eq.) was then added to the solution and the mixture was stirred at 120° C. in a sealed tube for 16 hr. The reaction was concentrated in vacuo and the residue was purified by column chromatography (SiO 2 , petroleum ether / ethyl acetate = 10: 1 to 3: 1) to give 7-bromo-12-(4-fluorophenyl)-9H-benzo [e] pyrrolo [1, 2-a] [1, 2, 4] triazolo [3, 4-c] [1, 4] diazepine (2.3 g, 3.4 mmol) as a yellow solid. 1H NMR (400 MHz, DMSO-d 6 ) δ = 9.22 (s, 1H), 7.95 (s, 1H), 7.80 (d, J = 8.4 Hz, 1H), 7.69 (d, J = 8.8 Hz, 1H), 7.61 (t, J = 6.0 Hz, 2H), 7.53 (s, 1H), 7.17 (t, J = 8.4 Hz, 2H), 7.11 (s, 1H), 5.29 (s, 2H).

[0570]

[0571] (S)-1-(12-(4-Fluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)-N,N-dimethylpyrrolidin-3-amine, 338. Synthesized using general procedure A with 7-bromo-12-(4-fluorophenyl)-9H-benzo[e]-pyrrolo[1,2-a]-[1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (S)-N,N-dimethylpyrrolidin-3-amine in place of tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 9.06 (s, 1H), 7.61-7.49 (m, 3H), 7.43 (s, 1H), 7.13 (s, 1H), 7.06 (t, J = 8.6Hz, 2H), 6.89 (br.s., 1H), 6.79 (d, J = 8.4Hz, 1H) ,5.17(s,2H),4.17-4.01(m,1H),3.87-3.75(m,1H),3.72-3.56(m,2H), 3.44(q,J=8.3Hz,1H),2.99(s,6H),2.60(br.s.,1H),2.40-2.22(m,1H). ESI[M+H]=429.2

[0572] Plan 32

[0573]

[0574] Chemical experimental methods:

[0575]

[0576] 7-Bromo-N-(2,2-dimethoxyethyl)-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11-amine, S115. To a solution of 7-bromo-11-chloro-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine (3.0 g, 8.0 mmol, 1.0 eq.) in THF (30 mL) and dioxane (30 mL) was added 2,2-dimethoxyethylamine (8.5 g, 80.8 mmol, 10.0 eq.) and the mixture was stirred in a sealed tube at 130° C. for 16 hr. The mixture was poured into 1N glacial HCl solution (300 mL) and extracted with ethyl acetate (300 mL*3). The combined organic phases were washed with brine (400 mL), dried over anhydrous Na2SO4, filtered and concentrated to yield 7-bromo-N-(2,2-dimethoxyethyl)-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11-amine (3.70 g, crude) as a dark brown solid, which was used without further purification. ESI [M+H] = 458.1 / 460.1

[0577]

[0578]

[0145] 7-Bromo-12-(4-fluorophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine, S116. A solution of 7-bromo-N-(2,2-dimethoxyethyl)-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11-amine (3.7 g, 8.0 mmol, 1.0 eq.) in dioxane (50 mL) and 1 M HCl (50 mL) was stirred at 70 °C for 16 hr and then concentrated. The residual solid was washed with methanol (30 mL) and filtered to give 7-bromo-12-(4-fluorophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine (2.5 g, 6.3 mmol, 78.6% yield) as a dark brown solid which was used directly without any purification. 1 H NMR (400MHz, DMSO-d6) δ = 8.22 (s, 1H), 7.97 (s, 1H), 7.84 (d, J = 5.6Hz, 2H), 7.66 (d, J = 8.0Hz, 2H), 7.59-7.55 (m, 2H), 7.21-7.18 (m, 3H), 5.38 (s, 2H). ESI[M+H]=394.1 / 396.1

[0579]

[0580]

[0145] 12-(4-Fluorophenyl)-7-(2,6-diazaspiro[3.4]octan-6-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine, S376. Synthesized using general procedure A, substituting 7-bromo-12-(4-fluorophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine-12-yl)benzonitrile and tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, chloroform-d) δ = 7.42 (dd, J = 5.5, 8.6Hz, 2H), 7.30 (s, 1H), 7.24-7.18 (m, 2H), 7.05-6.92 (m, 4H), 6.57- 6.46(m,2H),4.93(br.s.,2H),3.70-3.54(m,4H),3.50(s,2H),3.34(t,J=6.6Hz,2H),2.24(t,J=6.6Hz,2H). ESI[M+H]=426.1

[0581]

[0582] The general procedure is as follows:

[0583] 2-(6-(12-(4-fluorophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopentan-7-yl)-2,6-diazaspiro[3.4]octan-2-yl)ethanol, S369. To 12-(4-fluorophenyl)-7-(2,6-diazaspiro[3.4]octan-6-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopentan-7-yl)-2,6-diazaspiro[3.4]octan-2-yl)ethanol To a solution of pyroluolo[1,2-a][1,4]diazepine (16 mg, 37 umol, 1.0 eq.) and 2-bromoethanol (14.1 mg, 112.8 umol, 8.0 uL, 3.0 eq.) in CH 3 CN (3.0 mL) was added Na 2 CO 3 (7.9 mg, 75.2 umol, 2.0 eq.) and the mixture was stirred at 60° C. for 16 hr. The mixture was concentrated and the residue was purified by neutral preparative HPLC to give 2-(6-(12-(4-fluorophenyl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine-7-yl)-2,6-diazaspiro[3.4]octan-2-yl)ethanol (2.4 mg, 4.7 umol, 12.6% yield) as a white solid. 1H NMR (400MHz, methanol-d4) δ=7.56-7.48(m,3H),7.32(d,J=8.4Hz,1H),7.26(s,1H),7.17(s,1H),7.02(t,J=8.8Hz,2H),6.89(d,J=0.9Hz,1H),6.70( s,1H),6.62(d,J=8.8Hz,1H),5.01(s,2H),3.58(t,J=5.7Hz,2H),3.47( s, 2H), 3.42-3.32 (m, 6H), 2.69 (t, J = 5.7Hz, 2H), 2.22 (t, J = 6.6Hz, 2H). ESI[M+H]=470.3

[0584] Plan 33

[0585]

[0586] Chemical experimental methods:

[0587]

[0588] 7-Bromo-12-(4-fluorophenyl)-9H-benzo[e]pyrrolo[1,2-a]tetrazo[5,1-c][1,4]diazepine, S118. To a solution of 7-bromo-11-chloro-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine (2.1 g, crude) in anhydrous DMF (10 mL) was added TMSN3 (1.86 g, 16.17 mmol, 3.0 eq.) dropwise at 20°C. The reaction mixture was stirred at 20°C for 16 hr and poured into ice water (50 mL). The resulting mixture was extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (30 mL), dried over MgSO4, and concentrated. The residual solid was washed with MeOH (20 mL) and dried to give 7-bromo-12-(4-fluorophenyl)-9H-benzo[e]pyrrolo[1,2-a]tetrazo[5,1-c][1,4]diazepine (1.6 g, crude) as a gray solid, which was used directly without further purification. ESI [M+H] = 396.0 / 398.0

[0589]

[0590] 7-(cis-3,5-Dimethylpiperazin-1-yl)-12-(4-fluorophenyl)-9H-benzo[e]pyrrolo[1,2-a]tetrazo[5,1-c][1,4]diazepine, 395. Synthesized using general procedure A substituting 7-bromo-12-(4-fluorophenyl)-9H-benzo[e]pyrrolo[1,2-a]tetrazo[5,1-c][1,4]diazepine for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine-12-yl)benzonitrile and cis-2,6-dimethylpiperazine for tert-butyl piperazine-1-carboxylate. 1 HNMR (400MHz, methanol-d4) δ = 7.88 (d, J = 9.3Hz, 1H), 7.56 (dd, J = 5.5, 8.2Hz, 2H), 7.49 (s, 1H), 7.31 (br.s., 1H), 7.28-7.20 (m, 2H) ,7.07(t,J=8.6Hz,2H),5.26(s,2H),4.08(d,J=13.2Hz,2H),3.53(br.s.,2H),2.82(t,J=12.3Hz,2H),1.42(d,J=6.6Hz,6H). ESI[M+H]=430.2

[0591] Plan 34

[0592]

[0593]

[0594] Chemical experimental methods:

[0595]

[0596] 10-Amino-7-bromo-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one, S119. To a solution of 7-bromo-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one (1.0 g, 2.69 mmol, 1.0 eq.) in DMF (20 mL) was added NaH (161.64 mg, 4.04 mmol, 60% purity, 1.5 eq.) at 0°C. After stirring for 1 hr, (aminooxy)diphenylphosphine oxide (753.88 mg, 3.23 mmol, 1.2 eq.) was added. The reaction mixture was stirred at 30°C for 1 hr and then quenched by saturated NH4Cl solution (100 mL). The resulting mixture was extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL), dried over MgSO4 and concentrated. The residual solid was washed with TBME (20 mL) and dried to produce 10-amino-7-bromo-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one (700 mg, 1.81 mmol, 67.29% yield) as a yellow solid. ESI[M+H]=386.1 / 388.1

[0597]

[0598]

[0145] 7-Bromo-12-(4-fluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazepine, S120. A mixture of 10-amino-7-bromo-2-(4-fluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one (700 mg, 1.8 mmol, 1.0 eq.) in formamide (10 mL) was stirred at 200 °C for 2 hr and then poured into cold water (20 mL). The resulting precipitate was collected by filtration and purified by column chromatography on silica gel (petroleum ether / ethyl acetate = 20:1 to 2:1) to give 7-bromo-12-(4-fluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazepine (240 mg, 607.24 umol, 33.55% yield) as a yellow solid. TLC: R f =0.7 (petroleum ether / EtOAc=2 / 1). ESI[M+H]=394.9 / 396.9

[0599]

[0600] 1-(12-(4-Fluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazepin-7-yl)-N,N-dimethylpiperidin-4-amine, 428. Synthesized using general procedure A, substituting 7-bromo-12-(4-fluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazepin-7-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and N,N-dimethylpiperidin-4-amine for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 8.19 (s, 1H), 7.72 (d, J = 9.3Hz, 1H), 7.54 (dd, J = 53, 8.4Hz, 2H), 7.41 (s, 1H), 7.20-7.01 (m, 5H), 5.17 (s, 2H) ,4.04(d,J=13.2Hz,2H),3.47-3.35(m,1H),2.96-2.90(m,2H),2.89(s,6H),2.18(d,J=11.5Hz,2H),1.82(dq,J=3.7,12.1Hz,2H). ESI[M+H]=443.2

[0601] Plan 35

[0602]

[0603] Chemical experimental methods:

[0604]

[0605] Methyl 1-((3-amino-6-bromopyridin-2-yl)methyl)-4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate, S121. To a mixture of (3-amino-6-bromopyridin-2-yl)methanol (8.0 g, 39.40 mmol, 1.0 eq.) and EtN (12.00 g, 118.59 mmol, 16.44 mL, 3.01 eq.) in anhydrous DCM (1.60 L) and anhydrous THF (160 mL) was added dropwise MsCl (6.0 g, 52.40 mmol, 4.05 mL, 1.33 eq.) at -70°C under N2 atmosphere. The mixture was slowly warmed to 20°C and stirred for 30 min. The mixture was then cooled to 0°C again and methyl 4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate (5.5 g, 25.09 mmol, 0.64 eq.) was added, followed by BuNOH (5.0 g, 4.82 mmol, 6.25 mL, 25% in H0, 0.12 eq.), and NaOH solution (25% in H0, 30 g, 187.54 mmol, 4.76 eq.). The reaction mixture was slowly warmed to 25°C and stirred for an additional 15.5 hr. The mixture was washed with water (1 L) and brine (1 L), dried over Na0, and concentrated. The residue was purified by column chromatography on silica gel (petroleum ether / THF=30 / 1 to 5 / 1) to give methyl 1-((3-amino-6-bromopyridin-2-yl)methyl)-4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate (3.3 g, 6.94 mmol, 17.61% yield, 90% purity) as a yellow solid. TLC: R f =0.35 (petroleum ether / THF=5 / 1). 1 H NMR (400MHz, DMSO-d6) δ = 7.64-7.60 (m, 2H), 7.55 (br.s, 1H), 7.27 (br.s, 1H), 7.17-7.14(m,3H),6.99-6.97(m,1H),5.56(s,2H),5.43(s,2H),3.71(s,3H). ESI[M+H]=404.0 / 406.0

[0606]

[0607] 2-Bromo-8-(4-fluorophenyl)-5H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine-6(11H)-one, S122. To a suspension of methyl 1-((3-amino-6-bromopyridin-2-yl)methyl)-4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate (3.3 g, 8.16 mmol, 1.0 eq.) in anhydrous toluene (150 mL) was added dropwise AlMe (2 M in toluene, 20 mL, 4.9 eq.) at 0°C under N2 atmosphere. The reaction mixture was warmed to 25°C and stirred for 16 hours. The mixture was poured into cold aqueous NH4Cl solution (200 mL) and extracted with hot EtOAc / THF (4:1, 100 mL x 3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, and concentrated. The residual solid was washed with EtOH (10 mL x 2) and dried to give 2-bromo-8-(4-fluorophenyl)-5H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine-6(11H)-one (1.80 g, 4.05 mmol, 49.68% yield, 83.82% purity) as a light yellow solid. 1 H NMR (400MHz, DMSO-d6) δ = 10.35 (s, 1H), 7.69-7.64 (m, 2H), 7.59-7.54 (m, 3H), 7.19-7.14 (m, 3H), 5.34 (s, 2H). ESI[M+H]=372.0 / 374.0

[0608]

[0609] 2-Bromo-6-chloro-8-(4-fluorophenyl)-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine, S123.

[0610] To a suspension of 2-bromo-8-(4-fluorophenyl)-5H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine-6(11H)-one (600 mg, 1.61 mmol, 1.0 eq.) in anhydrous dioxane (20 mL) was added POCl (1.0 g, 6.52 mmol, 600 uL, 4.0 eq.) dropwise at 25° C. under N atmosphere. The mixture was stirred at 85° C. for 4 hours and then concentrated to dryness. The crude product was used directly without purification.

[0611]

[0612] 2-Bromo-N-(2,2-dimethoxyethyl)-8-(4-fluorophenyl)-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine-6-amine, S124. To a solution of 2-bromo-6-chloro-8-(4-fluorophenyl)-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine (600 mg, 1.6 mmol, 1.0 eq.) in anhydrous dioxane (50 mL) was added 2,2-dimethoxyethylamine (7 g, 66 mmol, 7.2 mL, 41 eq.) dropwise at 0° C. under N2 atmosphere. The mixture was stirred at 100° C. for 8 h and concentrated. The residue was purified by column chromatography on silica gel using petroleum / THF = 40:1 to 10:1 to give 2-bromo-N-(2,2-dimethoxyethyl)-8-(4-fluorophenyl)-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine-6-amine (600 mg, 1.09 mmol, 67.6% yield, 83.4% purity) as a light yellow solid, which was used directly without further purification. TLC: R f =0.6, petroleum ether / THF=2 / 1. ESI[M+H]=459.1 / 461.1

[0613]

[0614]

[0145] 7-Bromo-12-(4-fluorophenyl)-9H-imidazo[2,1-c]pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine, S125. A mixture of 2-bromo-N-(2,2-dimethoxyethyl)-8-(4-fluorophenyl)-11H-pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine-6-amine (700 mg, 1.27 mmol, 1.00 eq.) in dioxane (30 mL) and 1 M HCl (30 mL) was heated to 70 °C for 16 hr and then concentrated. The residual solid was washed with THF (10 mL x 2) and dried in vacuo to yield 7-bromo-12-(4-fluorophenyl)-9H-imidazo[2,1-c]pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine (500 mg, crude) as a light yellow solid, which was used directly without further purification. ESI [M+H] = 395.0 / 397.0

[0615]

[0616] 12-(4-Fluorophenyl)-7-(2,6-diazaspiro[3.4]octan-6-yl)-9H-imidazo[2,1-c]pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine, 440. Synthesized using general procedure A, substituting 7-bromo-12-(4-fluorophenyl)-9H-imidazo[2,1-c]pyrido[3,2-e]pyrrolo[1,2-a][1,4]diazepine for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine-12-yl)benzonitrile and tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 8.01 (s, 1H), 7.88 (d, J = 8.8Hz, 1H), 7.78 (s, 1H), 7.64 (s, 1H), 7.59 (t, J = 6.2Hz, 2H), 7.24 (s, 1H), 7.10 (t, J = 8.1Hz, 2H), 6.68 (d, J = 9.0Hz, 1H), 5.32 (s, 2H), 4.23-4.15 (m, 2H), 4.13-4.05 (m, 2H), 3.85 (br.s., 2H), 3.70-3.56 (m, 2H), 2.40 (t, J = 6.7Hz, 2H). ESI[M+H]=427.1

[0617] Plan 36

[0618]

[0619] Chemical experimental methods:

[0620]

[0621] Methyl 1-((5-amino-2-chloropyridin-4-yl)methyl)-4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate, S127. To a solution of (5-amino-2-chloropyridin-4-yl)methanol (4.0 g, 25 mmol, 1.0 eq.) and TEA (7.6 g, 75.6 mmol, 10.4 mL, 3.0 eq.) in DCM (400 mL) was added methanesulfonyl chloride (3.4 g, 30.2 mmol, 2.3 mL, 1.2 eq.) at -78°C under N2. The mixture was warmed to 26°C and stirred at 26°C under N2 for 1 hr. To the solution was added methyl 4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate (4.4 g, 20.1 mmol, 0.8 eq.), NaOH (25%, 6.00 eq.), and TBAOH (2.62 g, 2.52 mmol, 3.28 mL, 25% purity, 0.10 eq.) at 0°C. The mixture was stirred at 26°C for 16 hours, diluted with water (500 mL), and extracted with DCM (500 mL*2). The combined organic layers were dried over anhydrous MgSO4, filtered, and concentrated to yield a residue. The residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 5 / 1 to 1 / 1) to afford methyl 1-((5-amino-2-chloropyridin-4-yl)methyl)-4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate (1.9 g, 5.28 mmol, 20.94% yield) as a yellow solid. TLC: Rf = 0.22 (petroleum ether / ethyl acetate = 1 / 1). ESI [M+H] = 360.0

[0622]

[0623] 3-Chloro-8-(4-fluorophenyl)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine-10(11H)-one, S128. To a solution of methyl 1-((5-amino-2-chloropyridin-4-yl)methyl)-4-(4-fluorophenyl)-1H-pyrrole-2-carboxylate (1.8 g, 5.2 mmol, 1.0 eq.) in toluene (100 mL) was added dropwise AlMe (2 M, 12.85 mL, 5.0 eq.) at 0°C under N2. The mixture was then stirred at 20°C for 16 hr, quenched with ice water (200 mL) and extracted with hot EtOAc / THF (1 / 1, 200 mL*2). The combined organic layers were washed with brine (100 mL), dried over Na2SO4 and concentrated. The residual solid was washed with MeOH (20 mL). After filtration, the filter cake was collected to yield 3-chloro-8-(4-fluorophenyl)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine-10(11H)-one (1.60 g, crude) as a yellow solid. 1H NMR (400 MHz, methanol-d4) δ = 8.13 (s, 1H), 7.49-7.39 (m, 3H), 7.28 (d, J = 1.8 Hz, 1H), 7.14 (d, J = 1.8 Hz, 1H), 6.96 (t, J = 8.8 Hz, 2H), 5.16 (s, 2H). ESI [M+H] = 327.9

[0624]

[0625] 3,10-Dichloro-8-(4-fluorophenyl)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine, S129. A solution of 3-chloro-8-(4-fluorophenyl)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine-10(11H)-one (300 mg, 806 umol, 1.0 eq.) in POCl (5 mL) was stirred at 90° C. for 1 hr. The reaction mixture was concentrated under reduced pressure, diluted with THF (5 mL), adjusted to pH 7-8 with TEA and concentrated under reduced pressure to yield crude 3,10-dichloro-8-(4-fluorophenyl)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine (310 mg) without further purification.

[0626]

[0627] 3-Chloro-N-(2,2-dimethoxyethyl)-8-(4-fluorophenyl)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine-10-amine, S130. A solution of 3,10-dichloro-8-(4-fluorophenyl)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine (310 mg, 793 umol, 1.0 eq.) and 2,2-dimethoxyethylamine (834 mg, 7.94 mmol, 860 uL, 10 eq.) in THF (10 mL) / 1,4-dioxane (10 mL) was stirred in a sealed tube at 130° C. for 16 hr. The reaction mixture was concentrated under reduced pressure, diluted with THF / EtOAc (50 mL / 50 mL), and washed with 1 M HCl (20 mL). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give crude 3-chloro-N-(2,2-dimethoxyethyl)-8-(4-fluorophenyl)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazopentan-10-amine (360.00 mg, crude) as a dark brown oil without further purification.

[0628]

[0629]

[0145] 7-Chloro-12-(4-fluorophenyl)-9H-imidazo[2,1-c]pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine, S131. A solution of 3-chloro-N-(2,2-dimethoxyethyl)-8-(4-fluorophenyl)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine-10-amine (360 mg, 867 umol, 1.0 eq.) in dioxane (10 mL) and 1 M HCl (10 mL) was stirred at 70 °C for 16 hr and concentrated under reduced pressure. The residual solid was washed with THF (5 mL) and filtered to give crude 7-chloro-12-(4-fluorophenyl)-9H-imidazo[2,1-c]pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine (250 mg, crude) as a dark brown solid without further purification. ESI [M+H] = 350.9

[0630]

[0631] (R)-2-((((R)-1-(12-(4-fluorophenyl)-9H-imidazo[2,1-c]pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepin-7-yl)pyrrolidin-3-yl)methyl)amino)propan-1-ol, 442. The general procedure A was used with 7-chloro-12-(4-fluorophenyl)-9H-imidazo[2,1-c]pyridine The product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazol-12-yl)benzonitrile with 1H-benzo[3,4-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazol-12-yl)benzonitrile and (R)-2-(((S)-pyrrolidin-3-ylmethyl)amino)propan-1-ol with tert-butyl piperazine-1-carboxylate. NMR (400 MHz, methanol-d4) δ = 8.43 (s, 1H), 8.09 (br.s., 1H), 7.78 (br.s., 1H), 7.65-7.53 (m, 3H), 7.24 (br.s., 1H), 7.10 (t, J = 8.0 Hz, 2H), 6.76 (s, 1H), 5.33 (s, 2H), 3.94-3 .79(m,2H),3.72(br.s.,1H),3.65-3.50(m,2H),3.41(br.s.,2H),3.26-3.16(m,2 H),2.81-2.67(m,1H),2.36(br.s.,1H),2.02-1.86(m,1H),1.35(d,J=6.7Hz,3H). ESI[M+H]=473.1

[0632] Plan 37

[0633]

[0634] Chemical experimental methods:

[0635]

[0636] 3-Chloro-8-(4-fluorophenyl)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine-10(11H)-thione, S132. To a solution of 3-chloro-8-(4-fluorophenyl)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine-10(11H)-one (1.3 g, 3.9 mmol, 1.0 eq.) in THF (30 mL) was added Lawesson's reagent (2.1 g, 5.2 mmol, 1.3 eq.) and the mixture was stirred at 70 °C for 1 hr. The mixture was concentrated and the residual solid was washed with methanol (10 mL) to give 3-chloro-8-(4-fluorophenyl)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine-10(11H)-thione as a yellow solid (1.0 g, 2.9 mmol, 73.5% yield), which was used without any purification. ESI [M+H] = 344.1

[0637]

[0638] 3-Chloro-8-(4-fluorophenyl)-10-(methylthio)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine, S133. To a solution of 3-chloro-8-(4-fluorophenyl)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine-10(11H)-thione (1.0 g, 2.9 mmol, 1.0 eq.) in DMF (8.0 mL) was added K2CO3 (1.2 g, 8.7 mmol, 3.0 eq.) and iodomethane (2.0 g, 14 mmol, 905 uL, 5.0 eq.). The mixture was stirred at 26°C for 1 hr and filtered. The filtrate was concentrated under reduced pressure to give 3-chloro-8-(4-fluorophenyl)-10-(methylthio)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine (1.0 g, 2.8 mmol, 96% yield) as a yellow solid, which was used directly without any purification. ESI [M+H] = 358.1

[0639]

[0640]

[0145] 7-Chloro-12-(4-fluorophenyl)-9H-pyrido[3,4-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine, S134. To a solution of 3-chloro-8-(4-fluorophenyl)-10-(methylthio)-5H-pyrido[3,4-e]pyrrolo[1,2-a][1,4]diazepine (1.0 g, 2.8 mmol, 1.0 eq.) in n-BuOH (30 mL) was added formic hydrazide (837 mg, 14 mmol, 5.0 eq.) and the mixture was stirred at 120 °C for 16 hr. The mixture was concentrated and the residual solid was washed with 1N HCl (20 mL) to give 7-chloro-12-(4-fluorophenyl)-9H-pyrido[3,4-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine (650 mg, 1.85 mmol, 66.23% yield) as a yellow solid. ESI [M+H] = 352.2

[0641]

[0642] (S)-1-(1-(12-(4-Fluorophenyl)-9H-pyrido[3,4-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)pyrrolidin-3-yl)-N,N-dimethylmethanamine, 443. The general procedure C was used with 7-chloro-12-(4-fluorophenyl)-9H-pyrido[3,4-e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)pyrrolidin-3-yl)-N,N-dimethylmethanamine. ][1,2,4]triazolo[3,4-c][1,4]diazepine was used instead of 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine-12-yl)benzonitrile and (R)-(pyrrolidin-3-ylmethyl)carbamic acid tert-butyl ester was used instead of (azetidin-3-ylmethyl)(methyl)carbamic acid tert-butyl ester. 1 H NMR (400MHz, methanol-d4) δ = 9.04 (br.s., 1H), 8.35 (br.s., 1H), 7.61-7.51 (m, 2H), 7.4 2(s,1H),7.17(br.s.,1H),7.06(t,J=8.6Hz,2H),6.76(s,1H),5.18(br.s.,2H), 3.94-3.82(m,1H),3.69(d,J=6.7Hz,1H),3.58-3.47(m,1H),3.37-3.32(m,2H),2 .96(s,6H),2.90-2.76(m,1H),2.70(s,1H),2.34(br.s.,1H),1.96-1.83(m,1H). ESI[M+H]=444.2

[0643] Plan 38

[0644]

[0645] Chemical Experimental Methods

[0646]

[0647] 1-tert-Butyl 2-methyl 4-(2,4-difluorophenyl)-1H-pyrrole-1,2-dicarboxylate, S137. To a mixture of 1-tert-butyl 2-methyl 4-bromo-1H-pyrrole-1,2-dicarboxylate (34 g, 111 mmol, 1.0 eq.), (2,4-difluorophenyl)boronic acid (26 g, 164 mmol, 1.47 eq.), Boc2O (38 g, 174 mmol, 40 mL, 1.56 eq.), and Na2CO3 (24 g, 226 mmol, 2.0 eq.) in dioxane (1.0 L) and H2O (100 mL) was added Pd(dppf)Cl2 (4 g, 5.4 mmol, 0.05 eq.) at 25° C. under N2 atmosphere. The mixture was stirred at 100° C. for 16 hr and concentrated. The residue was diluted with EtOAc / THF (400 mL / 100 mL) and washed with brine (400 mL). The aqueous layer was separated and extracted with EtOAc (150 mL×2). The combined organic layers were dried over Na2SO4 and concentrated to produce a mixture of 1-tert-butyl 2-methyl 4-(2,4-difluorophenyl)-1H-pyrrole-1,2-dicarboxylate (40 g crude) as a dark brown oil, which was used directly. ESI[M+H]=338.0

[0648]

[0649] Methyl 4-(2,4-difluorophenyl)-1H-pyrrole-2-carboxylate, S138. To a solution of 1-tert-butyl 2-methyl 4-(2,4-difluorophenyl)-1H-pyrrole-1,2-dicarboxylate (40 g, crude) in DCM (50 mL) was added TFA (100 mL) dropwise. The reaction mixture was stirred at 25 °C for 1.0 hr and concentrated. MeOH (50 mL) was added to the residue and stirred for 1.0 hr. The solid was collected by filtration to give methyl 4-(2,4-difluorophenyl)-1H-pyrrole-2-carboxylate (15 g, 56.91 mmol, 56.41% yield over all 2 steps) as a dark brown solid, which was used directly. 1H NMR (400MHz, CDCl3) δ = 9.21, (br.s, 1H), 7.54-7.48, (m, 1H), 7.38 (s, 1H), 7.22 (s, 1H), 6.92-6.85 (m, 2H), 3.90 (s, 3H). ESI[M+H]=238.0

[0650]

[0651] Methyl 1-(5-bromo-2-nitrobenzyl)-4-(2,4-difluorophenyl)-1H-pyrrole-2-carboxylate, S139. To a solution of (5-bromo-2-nitrophenyl)methanol (9.0 g, 38.7 mmol, 1.0 eq.) and EtN (6.0 g, 59.2 mmol, 8.20 mL, 1.5 eq.) in anhydrous DCM (300 mL) was added MsCl (5.2 g, 45.5 mmol, 3.5 mL, 1.2 eq.) dropwise at 0 °C under N2 atmosphere. The reaction mixture was stirred at 0°C for 1.0 hr, followed by the addition of methyl 4-(2,4-difluorophenyl)-1H-pyrrole-2-carboxylate (9.0 g, 37.9 mmol, 1.0 eq.), BuNOH (25% in H2O, 4.0 g, 3.8 mmol, 0.1 eq.), and aqueous NaOH (25%, 35 g, 218.9 mmol, 5.7 eq.). The reaction mixture was warmed to 25°C and stirred for an additional 16 hr. The mixture was washed with brine (200 mL), dried over Na2SO4, and concentrated. The residual solid was washed with EtOAc / petroleum ether (1:1, 50 mL x 2), filtered, and dried to yield methyl 1-(5-bromo-2-nitrobenzyl)-4-(2,4-difluorophenyl)-1H-pyrrole-2-carboxylate (14.50 g, crude) as a yellow solid, which was used directly. ESI[M+H]=450.9 / 452.9

[0652]

[0653] Methyl 1-(2-amino-5-bromobenzyl)-4-(2,4-difluorophenyl)-1H-pyrrole-2-carboxylate, S140.

[0654] A mixture of 1-(5-bromo-2-nitrobenzyl)-4-(2,4-difluorophenyl)-1H-pyrrole-2-carboxylic acid methyl ester (14.5 g, 32.1 mmol, 1.0 eq.), Fe (9.0 g, 161 mmol, 5.0 eq.) and NHCl (5.0 g, 93.5 mmol, 3.27 mL, 2.9 eq.) in THF (100 mL), EtOH (100 mL) and H0 (50 mL) was stirred at 100°C for 2.5 hr and then concentrated. The residue was dissolved in hot THF (2 L), filtered, and the filtrate was concentrated to give 1-(2-amino-5-bromobenzyl)-4-(2,4-difluorophenyl)-1H-pyrrole-2-carboxylic acid methyl ester (13 g, crude) as a yellow solid, which was used directly. ESI [M+H] = 420.9 / 422.9

[0655]

[0656] 7-Bromo-2-(2,4-difluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one, S141. To a suspension of methyl 1-(2-amino-5-bromobenzyl)-4-(2,4-difluorophenyl)-1H-pyrrole-2-carboxylate (13 g, 30.8 mmol, 1.0 eq.) in anhydrous toluene (150 mL) was added dropwise AlMe (2 M in toluene, 75 mL, 4.8 eq.) at 0°C under N2 atmosphere. The reaction mixture was stirred at 25°C for 16 hr and then slowly poured into cold 0.5 M HCl (100 mL). The aqueous layer was separated and extracted with EtOAc / THF (8:1, 100 mL x 3). The combined organic layers were washed with brine (150 mL), dried over Na2SO4, and concentrated. The residual solid was washed with petroleum ether / EtOAc (3:1, 50 mL x 2) and dried in vacuo to yield 7-bromo-2-(2,4-difluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one (7.5 g, crude) as a light yellow solid, which was used directly without further purification. ESI [M+H] = 388.9 / 390.9

[0657]

[0658] 7-Bromo-11-chloro-2-(2,4-difluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine, S142. A solution of 7-bromo-2-(2,4-difluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one (1.5 g, 3.8 mmol, 1.0 eq.) in POCl (50 mL) was heated to 90° C. for 6 hr and then concentrated in vacuo to give the crude product, which was used directly without purification.

[0659]

[0660] 7-Bromo-12-(2,4-difluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine, S143. To a mixture of 7-bromo-11-chloro-2-(2,4-difluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine (1.6 g, 3.9 mmol, 1.0 eq.) in anhydrous dioxane (50 mL) was added formic hydrazide (2.4 g, 39.9 mmol, 10.1 eq.) and the reaction mixture was stirred at 110° C. for 16 hr. The mixture was then cooled to 25° C. and poured into ice-cold water (150 mL) and extracted with hot EtOAc / THF (4:1, 100 mL×4). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, and concentrated. The residual solid was washed with EtOAc (50 mL x 2) and dried in vacuo to yield 7-bromo-12-(2,4-difluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine (1.0 g, crude) as an off-white solid, which was used directly. ESI [M+H] = 412.9 / 414.9

[0661]

[0662] (S)-1-(12-(2,4-difluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-7-yl)-N,N-dimethylpyrrolidin-3-amine, 449. Synthesized using general procedure A, substituting 7-bromo-12-(2,4-difluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (S)-N,N-dimethylpyrrolidin-3-amine for tert-butyl piperazine-1-carboxylate.1 H NMR (400 MHz, methanol-d4) δ = 8.97 (s, 1H), 7.68-7.60 (m, 1H), 7.51-7.46 (m, 2H), 7.15 (s, 1H), 6.99-6.90 (m, 2H), 6.88 (d, J = 2.6 Hz, 1H), 6.76 (dd, J = 2.4, 8.6 Hz, 1H), 5.16 (s, 2H), 3. 91(br.s.,1H),3.79-3.72(m,1H),3.65(dt,J=3.1,9.0Hz,1H),3.55(dd,J=6.6,10.1Hz ,1H),3.46-3.37(m,1H),2.88(s,6H),2.73-2.65(m,1H),2.26(dd,J=7.9,12.8Hz,1H). ESI[M+H]=447.1

[0663] Plan 39

[0664]

[0665] Chemical experimental methods:

[0666]

[0667] 10-Amino-7-bromo-2-(2,4-difluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one, S144. To a solution of 7-bromo-2-(2,4-difluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one (1.0 g, 2.5 mmol, 1.0 eq.) in anhydrous DMF (15 mL) was added NaH (260 mg, 6.50 mmol, 60% purity, 2.53 eq.) and the mixture was stirred at 0° C. for 30 min. (Aminooxy)diphenylphosphine oxide (797.1 mg, 3.4 mmol, 1.3 eq.) was then added portionwise and the reaction mixture was stirred at 25° C. for an additional 2.5 hr. The mixture was quenched with ice-cold saturated aqueous NH4Cl (50 mL) and extracted with EtOAc (50 mL×3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and concentrated. The residue was purified by column chromatography on silica gel using petroleum / EtOAc (10:1 to 1:1) to produce 10-amino-7-bromo-2-(2,4-difluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one (600 mg, 1.34 mmol, 52.14% yield, 90.2% purity) as an off-white solid. TLC: R f=0.55 (petroleum ether / EtOAc=1 / 1). ESI[M+H]=404.0 / 406.0

[0668]

[0669]

[0145] 7-Bromo-12-(2,4-difluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazepine, S145. A mixture of 10-amino-7-bromo-2-(2,4-difluorophenyl)-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one (250 mg, 618 umol, 1.0 eq.) and ZnCl (250 mg, 1.8 mmol, 85.9 uL, 2.9 eq.) in NHCHO (5.0 mL) was stirred at 200 °C for 1 hr and then the mixture was poured into water (50 mL). The resulting precipitate was collected by filtration and then purified by preparative TLC (petroleum ether / EtOAc=2:1) to give 7-bromo-12-(2,4-difluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazepine (125 mg, 453 umol, 36% yield, 75% purity) as a light yellow solid, which was used directly. ESI [M+H]=413.0 / 415.0

[0670]

[0671] (S)-1-(12-(2,4-Difluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazepine-7-yl)-N-methylpyrrolidin-3-amine, 451. Synthesized using general procedure A, substituting 7-bromo-12-(2,4-difluorophenyl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[5,1-c][1,4]diazepine for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine-12-yl)benzonitrile and tert-butyl (S)-methyl(pyrrolidin-3-yl)carbamate for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 8.16 (s, 1H), 7.73 (d, J = 8.8Hz, 1H), 7.70-7.60 (m, 1H) ,7.50(s,1H),7.17(s,1H),7.03-6.92(m,2H),6.89-6.78(m,2H),5.25-5.17 (m,2H),4.05-3.94(m,1H),3.75-3.56(m,3H),3.46(dt,J=5.7,9.0Hz,1H),2 .86-2.74(m,3H),2.55(dt,J=6.7,14.4Hz,1H),2.28(dt,J=5.3,13.3Hz,1H). ESI[M+H]=433.1

[0672] Plan 40

[0673]

[0674] Chemical experimental methods:

[0675]

[0676] A suspension of 1-tert-butyl 2-methyl 4-bromo-1H-pyrrole-1,2-dicarboxylate (44 g, 144 mmol, 1.0 eq.), phenylboronic acid (26.4 g, 217 mmol, 1.5 eq.), Na2CO3 (30.6 g, 289 mmol, 12 mL, 2.0 eq.) and (Boc)2O (47.3 g, 217 mmol, 49.8 mL, 1.5 eq.), Pd(dppf)Cl2 (5.3 g, 7.2 mmol, 0.05 eq.) in dioxane / H2O (1.5 L, 10:1) was degassed and then heated to 80° C. to 100° C. under N2 for 12 hr. The mixture was concentrated under reduced pressure and the residue was partitioned between ethyl acetate (1.0 L) and brine (200 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated to yield crude 1-tert-butyl 2-methyl 4-phenyl-1H-pyrrole-1,2-dicarboxylate as an off-white solid (43.00 g, crude), which was used in the next step without further purification. ESI [M+H] = 302.2

[0677]

[0678] Methyl 4-phenyl-1H-pyrrole-2-carboxylate, S148. A solution of 2-methyl 1-tert-butyl 4-phenyl-1H-pyrrole-1,2-dicarboxylate (43 g, 142.7 mmol, 1.0 eq.) in TFA (300 mL) was stirred at 50 °C for 2 hr and then concentrated under reduced pressure. MeOH (200 mL) was added to the residue and stirred for 1 hr. The precipitate was collected by filtration to yield methyl 4-phenyl-1H-pyrrole-2-carboxylate (20 g, crude) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ = 7.60 (d, J = 7.4Hz, 2H), 7.49 (dd, J = 1.6, 2.7Hz, 1H), 7.30 (t, J = 7.6Hz, 2H), 7.19-7.11 (m, 2H), 3.77 (s, 3H). ESI[M+H]=202.2

[0679]

[0680]

[0146] Methyl 1-(5-bromo-2-nitrobenzyl)-4-phenyl-1H-pyrrole-2-carboxylate, S149. To a solution of (5-bromo-2-nitrophenyl)methanol (12.0 g, 51.7 mmol, 1.00 eq.) and TEA (10.4 g, 103.4 mmol, 14.3 mL, 2.0 eq.) in DCM (300 mL) was added MsCl (6.5 g, 56.8 mmol, 4.4 mL, 1.1 eq.) at 0 °C and the mixture was stirred at 0 °C for 0.5 hr. To the solution was then added methyl 4-phenyl-1H-pyrrole-2-carboxylate (10.4 g, 51.7 mmol, 1.0 eq) at 0°C, followed by tetrabutylammonium hydroxide (5.37 g, 5.2 mmol, 6.7 mL, 25% w%, 0.10 eq.) and NaOH solution (25%, 5.0 eq.). The mixture was stirred at 20°C for 11 hours and then diluted with ice water (200 mL) and DCM (800 mL). The organic layer was separated, dried over anhydrous Na2SO4, filtered, and concentrated. EtOH (50 mL) was added to the residue and stirred for 1 hour. The precipitate was collected by filtration to produce methyl 1-(5-bromo-2-nitrobenzyl)-4-phenyl-1H-pyrrole-2-carboxylate (20 g) as an off-white solid. 1 H NMR (400 MHz, chloroform-d) δ = 8.01 (d, J = 8.8 Hz, 1H), 7.54 (d, J = 7.5 Hz, 3H), 7.41-7.34 (m, 3H), 7.25 (s, 1H), 7.22 (d, J = 1.8 Hz, 1H), 6.73 (s, 1H), 5.92 (s, 2H), 3.75 (s, 3H)

[0681]

[0682] Methyl 1-(2-amino-5-bromobenzyl)-4-phenyl-1H-pyrrole-2-carboxylate, S150. A suspension of methyl 1-(5-bromo-2-nitrobenzyl)-4-phenyl-1H-pyrrole-2-carboxylate (17 g, 40.9 mmol, 1.0 eq.), Fe (11.43 g, 204.70 mmol, 5.00 eq.) and NHCl (10.9 g, 204.7 mmol, 7.1 mL, 5.0 eq.) in EtOH (80 mL), H2O (40 mL) and THF (80 mL) was heated to 80°C for 5 hr. The mixture was concentrated to dryness, and hot THF (2 L) was added to the residue. The mixture was filtered and the filtrate was concentrated. EtOH (100 mL) was added to the residue and stirred for 1 hr. The precipitate was collected by filtration to give methyl 1-(2-amino-5-bromobenzyl)-4-phenyl-1H-pyrrole-2-carboxylate (13.00 g) as a yellow solid. ESI [M+H] = 385.1 / 387.1

[0683]

[0684] 7-Bromo-2-phenyl-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one, S151. To a suspension of methyl 1-(2-amino-5-bromobenzyl)-4-phenyl-1H-pyrrole-2-carboxylate (13 g, 33.74 mmol, 1.0 eq.) in toluene (150 mL) was added MeAl (2 M in toluene, 85 mL, 5.0 eq.) at 0°C and the mixture was stirred at 30°C for 10 hr. The mixture was poured into 1 M glacial HCl (300 mL) and extracted with hot EtOAc / THF (1:1, 300 mL*4). The organic layer was dried over anhydrous NaSO, filtered, and concentrated. To the residue was added MeOH (50 mL) and stirred for 1 hr. The precipitate was collected by filtration to give 7-bromo-2-phenyl-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one as a white solid (10 g). 1 H NMR (400MHz, DMSO-d6) δ = 10.29 (s, 1H), 7.73 (s, 1H), 7.63-7.52 (m, 4H), 7.39 (t, J = 7.5Hz, 2H), 7.26-7.17 (m, 3H), 5.27 (s, 2H)

[0685]

[0686] 7-Bromo-11-chloro-2-phenyl-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine, S152. A solution of 7-bromo-2-phenyl-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11(10H)-one (1.0 g, 2.83 mmol, 1.0 eq.) in POCl (50 mL) was stirred at 90° C. for 2 hr. The solution was concentrated under reduced pressure to give 7-bromo-11-chloro-2-phenyl-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine (1.2 g, crude) as a black oil, which was used in the next step without further purification.

[0687]

[0688] 7-Bromo-12-phenyl-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine, S153. A solution of 7-bromo-11-chloro-2-phenyl-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine (670 mg, 1.75 mmol, 1.0 eq.) and formic hydrazide (525 mg, 8.75 mmol, 5.0 eq.) in dioxane (20 mL) was stirred at 130° C. in a sealed tube for 12 hours. The mixture was concentrated and the residue was partitioned between hot EtOAc / THF (1:1, 200 mL) and HCl (3 M, 50 mL). The organic phase was washed with brine (15 mL), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. To the residue was added MeOH (10 mL) and stirred for 1 hr. The precipitate was collected by filtration to give 7-bromo-12-phenyl-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine (500 mg, 1.19 mmol, 68.16% yield) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ=9.21(s,1H),7.94(br.s.,1H),7.78(d,J=8.2Hz,1H),7.67(d,J=8.6Hz ,1H),7.59-7.52(m,3H),7.33(t,J=7.0Hz,2H),7.16(t,J=7.2Hz,1H),7.11(s,1H),5.28(s,2H).

[0689]

[0690] (R)-1-((1-(12-phenyl-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine-7-yl)pyrrolidin-3-yl)methyl)piperidin-4-ol, 459. Synthesized using General Procedure A with 7-bromo-12-phenyl-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine-12-yl)benzonitrile and (S)-1-(pyrrolidin-3-ylmethyl)piperidin-4-ol replacing tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 9.14 (br.s., 1H), 7.59-7.39 (m, 4H), 7.32 (t, J = 7.5Hz, 2H), 7.24 -7.12(m,2H),6.74(br.s.,1H),6.65(d,J=8.4Hz,1H),5.08(s,2H),4.11(br.s.,1H),3. 84(br.s.,1H),3.76-3.58(m,2H),3.54-3.42(m,2H),3.42-3.32(m,2H),3.17-3.02(m,2 H), 2.93-2.76 (m, 1H), 2.33 (d, J = 5.3Hz, 1H), 2.15 (d, J = 13.2Hz, 1H), 2.11-1.69 (m, 5H). ESI[M+H]=481.2

[0691] Plan 41

[0692]

[0693] Chemical experimental methods:

[0694]

[0695] 7-Bromo-N-(2,2-dimethoxyethyl)-2-phenyl-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11-amine, S154. To a solution of 7-bromo-11-chloro-2-phenyl-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine (1.05 g, 2.83 mmol, 1.0 eq.) in dioxane (30 mL) and THF (30 mL) was added TEA (1.4 g, 14.1 mmol, 1.9 mL, 5.0 eq.) and 2,2-dimethoxyethylamine (5.9 g, 56.6 mmol, 6.1 mL, 20.0 eq.). The mixture was stirred in a sealed tube at 130° C. for 16 hr and concentrated. The residue was diluted with solvent EtOAc (300 mL) and THF (100 mL), washed with 0.5 M HCl (100 mL), saturated NaHCO 3 (100 mL) and brine (100 mL), dried over Na 2 SO 4 , filtered and concentrated to give crude 7-bromo-N-(2,2-dimethoxyethyl)-2-phenyl-5H-benzo[e]pyrrolo[1,2-a][1,4]diazolidine-11-amine (1.25 g, crude) as a dark brown oil. ESI [M+H] = 440.0 / 442.0

[0696]

[0697] 7-Bromo-12-phenyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine, S155. A mixture of 7-bromo-N-(2,2-dimethoxyethyl)-2-phenyl-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-11-amine (1.25 g, 2.84 mmol, 1.0 eq.) in dioxane (20 mL) and 1 M HCl (20 mL) was stirred at 80° C. for 16 hr. After the reaction was complete, the mixture was concentrated and THF (15 mL) and ethanol (1 mL) were added to the residue. After stirring for 1 hr, the precipitate was collected by filtration to give 7-bromo-12-phenyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine (1.5 g, crude) as a dark brown solid. ESI [M+H] = 376.0 / 378.0

[0698]

[0699] 12-Phenyl-7-(2,6-diazaspiro[3.4]octan-6-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine, 462. Synthesized using general procedure A substituting 7-bromo-12-phenyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepine-12-yl)benzonitrile and tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate for tert-butyl piperazine-1-carboxylate. 1HNMR (400MHz, methanol-d4) δ=7.56-7.48(m,3H),7.38-7.25(m,4H),7.17(s,1H),7.15-7.08(m,1H),6.93(s,1H),6.73(b r.s.,1H),6.65(d,J=6.8Hz,1H),5.04(s,2H),3.86-3.69(m,2H),3.54(s,2H),3.49-3.34(m,4H),2.35-2.24(m,2H). ESI[M+H]=408.1

[0700] Amine synthesis

[0701]

[0702] Chemical experimental methods:

[0703]

[0704] tert-Butyl 6-oxa-3-azabicyclo[3.1.0]hexane-3-carboxylate, M2. To a solution of tert-butyl 2,5-dihydropyrrole-1-carboxylate (100 g, 590.9 mmol, 1.0 eq.) in DCM (1.2 L) was added portionwise m-CPBA (180 g, 886.43 mmol, 85% purity, 1.50 eq.) at 0° C. and the mixture was stirred at 25° C. for 16 hr. The mixture was washed with 10% aqueous NaHSO 3 solution (600 mL×2), saturated aqueous NaHCO 3 solution (600 mL×2), and brine (600 mL), dried over Na 2 SO 4, and concentrated to give tert-butyl 6-oxa-3-azabicyclo[3.1.0]hexane-3-carboxylate (220 g, crude) as a yellow oil. 1H NMR (400 MHz, chloroform-d) δ = 3.75 (d, J = 12.8 Hz, 1H), 3.67 (d, J = 12.8 Hz, 1H), 3.60 (d, J = 3.3 Hz, 2H), 3.24 (dd, J = 5.0, 12.7 Hz, 2H), 1.43-1.31 (m, 9H)

[0705]

[0706] tert-Butyl trans-3-(benzylamino)-4-hydroxypyrrolidine-1-carboxylate, M3. To a solution of tert-butyl 6-oxa-3-azabicyclo[3.1.0]hexane-3-carboxylate (135 g, 728.8 mmol, 1.0 eq.) in EtOH (1.3 L) was added portionwise BnNH2 (160 g, 1.49 mol, 163.27 mL, 2.05 eq.) at 25°C. The reaction mixture was heated to reflux (oil bath 95°C) for 16 hours and then concentrated. To the residue was added EtOAc / petroleum ether (1:1, 400 mL) and stirred for 1.0 hour. The precipitate was collected by filtration to yield tert-butyl trans-3-(benzylamino)-4-hydroxypyrrolidine-1-carboxylate (137 g, 421.7 mmol, 57.8% yield, 90% purity) as a white solid. 1H NMR (400 MHz, chloroform-d) δ = 7.37-7.18 (m, 5H), 4.09 (d, J = 4.0 Hz, 1H), 3.88-3.73 (m, 2H), 3.73-3.56 (m, 2H), 3.31-3.07 (m, 3H), 1.48-1.37 (m, 9H)

[0707] General Procedure K

[0708]

[0709] tert-Butyl (3S,4S)-3-(Benzylamino)-4-hydroxypyrrolidine-1-carboxylate, M4. To a suspension of tert-butyl trans-3-(Benzylamino)-4-hydroxypyrrolidine-1-carboxylate (151 g, 516.4 mmol, 1.0 eq.) in MeCN (4.20 L) and H₂O (280.00 mL) was added (2S)-2-hydroxy-2-phenyl-acetic acid (85.6 g, 562.9 mmol, 1.1 eq.). The mixture was heated to 85°C for 2 hours to slowly dissolve the solid. The solution was cooled to 20°C for 11 hours and the precipitate was collected by filtration. The filter cake was recrystallized again from MeCN / H₂O (3 L, 20:1) to yield another batch of solid. The solid was added to aqueous K₂CO₃ (3%, 1.0 L) and extracted with EtOAc (500 mL*3). The organic layer was dried over Na 2 SO 4 and concentrated to give tert-butyl (3S,4S)-3-(benzylamino)-4-hydroxypyrrolidine-1-carboxylate (50 g, 168 mmol, 32.7% yield, 98.7% purity) as a white solid. By SFC, ee% = 97.5%.

[0710]

[0711] tert-Butyl (3S,4S)-3-(dimethylamino)-4-hydroxypyrrolidine-1-carboxylate, M5. To a solution of tert-butyl (3S,4S)-3-(benzylamino)-4-hydroxypyrrolidine-1-carboxylate (30 g, 102 mmol, 1.0 eq.) and HCHO (102.7 g, 1.0 mol, 94.2 mL, 10 eq.) in MeOH (1.0 L) was added Pd(OH) (2.8 g, 10.2 mmol, 50% purity, 0.1 eq.) and the mixture was heated to 50° C. under 50 psi of hydrogen for 5 hr. The mixture was filtered and concentrated to give tert-butyl (3S,4S)-3-(dimethylamino)-4-hydroxypyrrolidine-1-carboxylate (23.5 g, 102 mmol, 99.4% yield) as a yellow oil. 1H NMR (400 MHz, chloroform-d) δ=4.23-4.20 (m, 1H), 3.75-3.50 (m, 2H), 3.26-3.14 (m, 2H), 2.75 (m, 1H), 2.29 (s, 6H), 1.42 (s, 9H)

[0712]

[0713] (3S,4S)-4-(Dimethylamino)pyrrolidin-3-ol, M6. A solution of tert-butyl (3S,4S)-3-(dimethylamino)-4-hydroxypyrrolidine-1-carboxylate (23.5 g, 102 mmol, 1.0 eq.) in HCl / MeOH (4 M, 200 mL) was stirred at 40°C for 2 hours. The mixture was concentrated to dryness. 100 mL of MeOH was then added to the residue and the solution was added to the basic resin. The mixture was stirred for 1 hour and the pH of the solution was adjusted to 7-8. The mixture was filtered and the filtrate was concentrated. 100 mL of THF was added to the residue and the suspension was stirred overnight. The solid was collected by filtration to yield (3S,4S)-4-(dimethylamino)pyrrolidin-3-ol (13 g, 99 mmol, 97.8% yield) as a yellow solid. 1H NMR (400 MHz, chloroform-d) δ=4.35 (br.s., 3H), 3.41-3.44 (m, 1H), 3.29-3.32 (m, 1H), 3.13-3.18 (m, 2H), 2.85 (m, 1H), 2.27 (s, 6H)

[0714]

[0715] General Procedure

[0716]

[0717] tert-Butyl (3S,4S)-3-amino-4-hydroxypyrrolidine-1-carboxylate, M7. To a solution of tert-butyl (3S,4S)-3-(benzylamino)-4-hydroxypyrrolidine-1-carboxylate (20 g, 68.4 mmol, 1.0 eq) in AcOH (300 mL) was added Pd(OH)2 (5.0 g, 3.56 mmol, 10% purity, 0.05 eq.) and the mixture was stirred at 50°C under 50 psi H2 for 1 hr. The mixture was filtered and the filtrate was concentrated to give tert-butyl (3S,4S)-3-amino-4-hydroxypyrrolidine-1-carboxylate (17.9 g, 68.3 mmol, 100% yield, AcOH salt) as a yellow oil. ESI [M+H] = 203.1

[0718]

[0719] To a solution of tert-butyl (3S,4S)-3-(((benzyloxy)carbonyl)amino)-4-hydroxypyrrolidine-1-carboxylate (29.0 g, 110.5 mmol, 1.0 eq., AcOH salt) in THF (290 mL) and H2O (290 mL) was added K2CO3 (45.8 g, 331 mmol, 3.0 eq.) and CbzCl (24.5 g, 143.7 mmol, 20.4 mL, 1.3 eq.) at 0°C. The reaction was stirred at 25°C for 1 hr and extracted with EtOAc (500 mL*3). The organic layer was washed with brine (500 mL), dried over MgSO4, and concentrated. The residue was purified by column chromatography (SiO 2 , petroleum ether / ethyl acetate = 10:1 to 1:1) to produce tert-butyl (3S,4S)-3-(((benzyloxy)carbonyl)amino)-4-hydroxypyrrolidine-1-carboxylate (18.3 g, 54.4 mmol, 49.2% yield) as a colorless oil. 1H NMR (400 MHz, methanol-d 4 ) δ = 7.32-7.26 (m, 5H), 5.06 (s, 2H), 4.08 (s, 1H), 4.07-3.89 (s, 1H), 3.65-3.61 (s, 1H), 3.50-3.48 (s, 1H), 3.29-3.22 (m, 2H), 1.43 (s, 9H). ESI [M+H] = 336.9

[0720]

[0721] To a solution of tert-butyl (3S,4R)-3-(((benzyloxy)carbonyl)amino)-4-((4-nitrobenzoyl)oxy)pyrrolidine-1-carboxylate (8.0 g, 23.7 mmol, 1.0 eq.), 4-nitrobenzoic acid (5.9 g, 35.6 mmol, 1.5 eq.), and PPh (11.2 g, 42.8 mmol, 1.8 eq.) in toluene (200 mL) was added DEAD (7.4 g, 42.8 mmol, 7.7 mL, 1.8 eq.) in one portion at 0° C. under N2. After stirring for 10 min, the mixture was heated to 80° C. for 16 hr. The solution was concentrated and purified by silica gel chromatography (petroleum ether / ethyl acetate = 3:1) to give tert-butyl (3S,4R)-3-(((benzyloxy)carbonyl)amino)-4-((4-nitrobenzoyl)oxy)pyrrolidine-1-carboxylate (14 g, crude) as a yellow solid. ESI [M+H] = 486.2

[0722]

[0723] tert-Butyl (3S,4R)-3-(((benzyloxy)carbonyl)amino)-4-hydroxypyrrolidine-1-carboxylate, M10. To a solution of tert-butyl (3S,4R)-3-(((benzyloxy)carbonyl)amino)-4-((4-nitrobenzoyl)oxy)pyrrolidine-1-carboxylate (6.0 g, 12.3 mmol, 1.0 eq.) in MeOH (20 mL) was added KCO (2.0 g, 14.8 mmol, 1.2 eq.) in one portion and the mixture was stirred at 20° C. for 10 min. The mixture was filtered and the filtrate was adjusted to pH=3 with 1 M HCl. The filtrate was concentrated and purified by column chromatography to give tert-butyl (3S,4R)-3-(((benzyloxy)carbonyl)amino)-4-hydroxypyrrolidine-1-carboxylate (4.1 g, crude) as a yellow oil.

[0724]

[0725] (3S,4R)-3-(Dimethylamino)-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester, M11. To a solution of (3S,4R)-3-(benzyloxycarbonylamino)-4-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester (3.6 g, 10.7 mmol, 1.0 eq.) and formaldehyde (6.4 g, 214 mmol, 5.9 mL, 20 eq.) in MeOH (30 mL) was added Pd(OH) (20.5 g, 14.6 mmol, 10% purity, 1.3 eq.) and the mixture was stirred under 50 psi H at 20° C. for 16 hr. The mixture was filtered and the filtrate was concentrated. The residue was purified by silica gel chromatography (DCM:MeOH=20:1) to yield tert-butyl (3S,4R)-3-(dimethylamino)-4-hydroxypyrrolidine-1-carboxylate (2.0 g, crude) as a yellow oil. 1H NMR (400 MHz, Methanol-d4) δ = 4.22 (br. s., 1H), 3.65-3.54 (m, 1H), 3.48-3.37 (m, 2H), 3.27-3.16 (m, 1H), 2.61-2.45 (m, 1H), 2.29 (d, J = 2.4 Hz, 6H), 1.56-1.36 (m, 9H). ESI [M+H] = 231.1

[0726]

[0727] (3R,4S)-4-(Dimethylamino)pyrrolidin-3-ol, M12. A solution of tert-butyl (3S,4R)-3-(dimethylamino)-4-hydroxypyrrolidine-1-carboxylate (1 g, 4.3 mmol, 1.0 eq.) in HCl / MeOH (4 M, 20 mL) was stirred at 20° C. for 30 min. The solution was concentrated to yield (3R,4S)-4-(dimethylamino)pyrrolidin-3-ol (600 mg, HCl salt) as a yellow oil. 1H NMR (400 MHz, Methanol-d4) δ = 4.70 (br. s., 1H), 4.13-4.01 (m, 1H), 3.88 (t, J = 10.2 Hz, 1H), 3.57-3.43 (m, 3H), 3.11-2.90 (m, 6H). ESI[M+H]=131.1

[0728]

[0729] Chemical experimental methods:

[0730]

[0731] cis-2-Benzyltetrahydro-2H-pyrrolo[3,4-d]isoxazole-5(3H)-carboxylic acid tert-butyl ester, M13. To a solution of tert-butyl 2,5-dihydropyrrole-1-carboxylate (70.6 g, 417.6 mmol, 1.0 eq.) in toluene (2.1 L) and EtOH (700 mL) was added (HCHO) n (62.8 g, 697 mmol, 1.6 eq.), N-benzylhydroxylamine (100 g, 626 mmol, 1.5 eq., HCl salt) and TEA (63.4 g, 626 mmol, 86 mL, 1.5 eq.). The mixture was stirred at 80 ° C for 24 hr and then concentrated under reduced pressure. The residue was diluted with hexane / EtOAc (1.5 L / 1.5 L), filtered and the filtrate was concentrated under reduced pressure to produce a yellow oily cis-2-benzyltetrahydro-2H-pyrrolo [3, 4-d] isoxazole-5 (3H) -carboxylic acid tert-butyl ester (230 g, crude), which was used in the next step without further purification. ESI [M + H] = 305.1

[0732]

[0733] cis-tert-Butyl 3-((Benzylamino)methyl)-4-hydroxypyrrolidine-1-carboxylate, M14.

[0734] tert-Butyl cis-2-benzyltetrahydro-2H-pyrrolo[3,4-d]isoxazole-5(3H)-carboxylate (229 g, 752 mmol, 1.0 eq.) was dissolved in AcOH (2.5 L). The solution was heated to 70°C and Zn powder (245 g, 3.7 mol, 5.0 eq.) was then added portionwise with vigorous stirring, followed by another batch of Zn powder (80 g) 30 minutes later. The mixture was stirred at 70°C for 1.5 hours, diluted with water (1.5 L), and extracted with DCM (2 L * 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO 2 , DCM:MeOH = 1 / 0 to 10:1) to yield cis-tert-butyl 3-((benzylamino)methyl)-4-hydroxypyrrolidine-1-carboxylate (107 g, 272 mmol, 36.2% yield, 78% purity) as a light orange solid. 1H NMR (400 MHz, CHLOROFORM-d) δ = 7.31-7.17 (m, 5H), 4.40-4.32 (m, 1H), 3.78-3.67 (m, 2H), 3.46-3.35 (m, 2H), 3.34-3.09 (m, 2H), 2.99-2.84 (m, 2H), 2.20 (br. s., 1H), 1.48-1.31 (s, 9H). ESI [M+H] = 307.0

[0735]

[0736] tert-Butyl cis-3-(aminomethyl)-4-hydroxypyrrolidine-1-carboxylate, M15. To a solution of tert-butyl cis-3-((benzylamino)methyl)-4-hydroxypyrrolidine-1-carboxylate (20 g, 65.2 mmol, 1.0 eq.) in MeOH (500 mL) was added Pd(OH)2 (13.9 g, 99 mmol, 1.5 eq.) and the mixture was stirred at 70°C under 50 psi H2 for 36 hr. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give crude tert-butyl cis-3-(aminomethyl)-4-hydroxypyrrolidine-1-carboxylate (11 g, crude) as a colorless oil, which was used in the next step without further purification. ESI [M+H] = 217.1

[0737]

[0738] tert-Butyl cis-3-(((benzyloxy)carbonyl)amino)methyl)-4-hydroxypyrrolidine-1-carboxylate, M16. To a solution of tert-butyl cis-3-(aminomethyl)-4-hydroxypyrrolidine-1-carboxylate (11 g, 50.8 mmol, 1.0 eq.) in H2O (50 mL) and THF (50 mL) was added K2CO3 (14 g, 101.7 mmol, 2.0 eq.) and CbzCl (10.4 g, 61 mmol, 8.6 mL, 1.2 eq.). The reaction was stirred at 20°C for 12 hr and then concentrated under reduced pressure. The residue was diluted with H2O (50 mL) and extracted with DCM (50 mL*3). The combined organic layers were concentrated and purified by column chromatography (SiO2, dichloromethane / ethyl acetate = 10 / 1 to 1 / 1 to DCM / methanol = 20 / 1) to give tert-butyl cis-3-((((benzyloxy)carbonyl)amino)methyl)-4-hydroxypyrrolidine-1-carboxylate (10 g) as a white solid.

[0739]

[0740] tert-Butyl (3S,4S)-3-((((Benzyloxy)carbonyl)amino)methyl)-4-hydroxypyrrolidine-1-carboxylate and tert-butyl (3R,4R)-3-((((Benzyloxy)carbonyl)amino)methyl)-4-hydroxypyrrolidine-1-carboxylate, M16A and M16B. cis-tert-Butyl 3-((((Benzyloxy)carbonyl)amino)methyl)-4-hydroxypyrrolidine-1-carboxylate was separated by SFC to yield tert-butyl (3S,4S)-3-(benzyloxycarbonylaminomethyl)-4-hydroxy-pyrrolidine-1-carboxylate (4.0 g, 10.7 mmol, 21% yield, 93.7% purity) as a white solid ( by SFC, ee% = 100%) and (3R,4R)-3-(benzyloxycarbonylaminomethyl)-4-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester (4.0 g, 10.10 mmol, 19.8% yield, 88% purity) as a white solid ( By SFC, ee% = 99.5%).

[0741] General Procedures

[0742]

[0743]

[0146] (3R,4R)-tert-Butyl 3-((Dimethylamino)methyl)-4-hydroxypyrrolidine-1-carboxylate, M17. A mixture of (3R,4R)-3-(benzyloxycarbonylaminomethyl)-4-hydroxy-pyrrolidine-1-carboxylate (2.0 g, 5.71 mmol, 1.0 eq.), HCHO (4.5 g, 57.1 mmol, 4.14 mL, 10 eq.), and Pd(OH) (1.6 g, 5.7 mmol, 50% purity, 1.0 eq.) in MeOH (50 mL) was stirred under 50 psi H at 50°C for 50 hr. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (SiO 2 , DCM / MeOH = 100 / 1 to 20: 1) to give tert-butyl (3R, 4R) -3- ((dimethylamino) methyl) -4-hydroxypyrrolidine-1-carboxylate (1.2 g, 4.9 mmol, 86% yield) as a white solid. 1H NMR (400 MHz, CHLOROFORM-d) δ = 4.43 (d, J = 5.5 Hz, 1H), 3.72-3.57 (m, 1H), 3.50-3.38 (m, 1H), 3.35-3.20 (m, 1H), 3.16-3.00 (m, 1H), 2.81 (q, J = 11.6 Hz, 1H), 2.45-2.27 (m, 8H), 1.44 (s, 9H). ESI [M + H] = 245.0

[0744]

[0745] (3R,4S)-4-((dimethylamino)methyl)pyrrolidin-3-ol, M18. A solution of tert-butyl (3R,4R)-3-((dimethylamino)methyl)-4-hydroxypyrrolidine-1-carboxylate (1.2 g, 4.9 mmol, 1.0 eq.) in HCl / MeOH (4 M, 20 mL) was stirred at 45 °C for 1 hr. The reaction mixture was concentrated under reduced pressure. The residue was diluted with HO (10 mL) and basified by passing through a basic resin. The mixture was filtered and the filtrate was lyophilized to yield (3R,4S)-4-[(dimethylamino)methyl]pyrrolidin-3-ol (700 mg, crude) as a light yellow solid. 1H NMR (400 MHz, methanol-d4) δ = 4.46 (t, J = 3.1 Hz, 1H), 3.53 (dd, J = 8.6, 11.3 Hz, 1H), 3.39-3.31 (m, 2H), 3.28 (br. s., 1H), 3.14-3.03 (m, 2H), 2.90 (dd, J = 6.7, 12.9 Hz, 1H), 2.63 (s, 6H). ESI [M+H] = 145.0

[0746]

[0747] General Procedure N

[0748]

[0749] (3R,4S)-tert-butyl 3-((((benzyloxy)carbonyl)amino)methyl)-4-((4-nitrobenzoyl)oxy)pyrrolidine-1-carboxylate, M19. To a solution of (3R,4R)-3-(benzyloxycarbonylaminomethyl)-4-hydroxy-pyrrolidine-1-carboxylate (1.0 g, 2.8 mmol, 1.0 eq.) in toluene (20 mL) was added 4-nitrobenzoic acid (714 mg, 4.2 mmol, 1.5 eq.) and PPh (1.5 g, 5.7 mmol, 2.0 eq.) at 25° C. under N. DEAD (992 mg, 5.7 mmol, 1.0 mL, 2.0 eq.) was then added at 0° C. and the mixture was heated to 80° C. under N for 16 hr. The reaction mixture was concentrated and the residue was purified by column chromatography (SiO 2 , petroleum ether / ethyl acetate = 20 / 1 to 3: 1) to give tert-butyl (3R, 4S)-3-((((benzyloxy)carbonyl)amino)methyl)-4-((4-nitrobenzoyl)oxy)pyrrolidine-1-carboxylate (2.2 g, crude) as a yellow oil. ESI [M + H] = 500.1

[0750]

[0751] To a solution of tert-butyl (3R,4S)-3-((((benzyloxy)carbonyl)amino)methyl)-4-hydroxypyrrolidine-1-carboxylate (2.2 g, 4.4 mmol, 1.0 eq.) in MeOH (30 mL) was added KCO (1.2 g, 8.8 mmol, 2.0 eq.) and the mixture was stirred at 25 °C for 30 min. The mixture was concentrated in vacuo. The residue was dissolved in water (30 mL) and extracted with DCM (20 mL*3). The organic layer was dried over MgSO and concentrated. The residue was purified by column chromatography (SiO 2 , petroleum ether / ethyl acetate = 20 / 1 to 1:1) to give (3R,4S)-3-(benzyloxycarbonylaminomethyl)-4-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester (900 mg, crude) as a yellow oil. 1 H NMR (400 MHz, methanol-d4) δ = 7.38-7.29 (m, 4H), 7.27 (d, J = 4.9 Hz, 1H), 5.06 (s, 2H), 4.08-3.99 (m, 1H), 3.57-3.46 (m, 2H), 3.21-3.10 (m, 3H), 3.07-2.98 (m, 1H), 2.23 (dd, J = 4.0, 7.1 Hz, 1H), 1.43 (s, 9H). ESI [M+H] = 351.2

[0752]

[0753] To a solution of (3R, 4S)-3-((dimethylamino)methyl)-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester (900 mg, 2.5 mmol, 1.0 eq.) in MeOH (50 mL) was added Pd(OH) (0.5 g) and the mixture was stirred at 60 ° C under 50 psi H for 16 hr. After filtration, the filtrate was concentrated. The residue was acidified by cold 0.5 M HCl (30 mL) and extracted with EtOAC (10 mL * 2). The aqueous phase was adjusted to pH = 11 with saturated K2CO3 solution and extracted with DCM (50 mL * 4). The organic layer was dried over MgSO 4 and concentrated to give tert-butyl (3R,4S)-3-((dimethylamino)methyl)-4-hydroxypyrrolidine-1-carboxylate (250 mg, crude) as a yellow oil, which was used in the next step without further purification. 1H NMR (400 MHz, methanol-d4) δ = 4.07-3.97 (m, 1H), 3.60 (dd, J = 5.3, 10.1 Hz, 1H), 3.52 (dd, J = 6.0, 11.2 Hz, 1H), 3.19-3.05 (m, 2H), 2.46-2.33 (m, 1H), 2.31-2.18 (m, 8H), 1.44 (s, 9H). ESI [M+H] = 245.1

[0754]

[0755] (3S,4S)-4-((Dimethylamino)methyl)pyrrolidin-3-ol, M22. To a solution of (3R,4S)-3-[(dimethylamino)methyl]-4-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester (250 mg, 1.02 mmol, 1.0 eq.) in MeOH (5.0 mL) was added HCl / MeOH (4 M, 10 mL) and the mixture was stirred at 25° C. for 30 min. The reaction was concentrated in vacuo. The residue was dissolved in MeOH (10 mL) and basified to pH = 7-8 by anion exchange resin. After filtration, the filtrate was concentrated to give (3S,4S)-4-[(dimethylamino)methyl]pyrrolidin-3-ol (150 mg, crude) as a yellow oil, which was used in the next step without further purification. 1 H NMR (400MHz, methanol-d4) δ = 4.23 (td, J = 2.6, 5.0Hz, 1H), 3.54 (dd, J = 7.1, 11.9Hz, 1H), 3.33 (dd, J = 4.9, 12.3Hz, 1H), 3.15(dd,J=2.6,12.3Hz,1H),3.06(dd,J=4.9,11.9Hz,1H),2.51-2.41(m,2H),2.40-2.35(m,1H),2.33(s,6H). ESI[M+H]=145.1

[0756]

[0757] General Procedure

[0758]

[0759]

[0266] trans-tert-Butyl 3-hydroxy-4-(((4-methoxybenzyl)(methyl)amino)methyl)pyrrolidine-1-carboxylate, M24. To a solution of tert-butyl trans-3-(aminomethyl)-4-hydroxy-pyrrolidine-1-carboxylate (2.0 g, 9.2 mmol, 1.0 eq.) in MeOH (40 mL) was added AcOH (666 mg, 11 mmol, 1.2 eq.) and 4-methoxybenzaldehyde (1.5 g, 11 mmol, 1.2 eq.). The mixture was stirred at 26°C for 0.5 h and then NaBHCN (1.4 g, 23.1 mmol, 2.5 eq) was added followed by HCHO (3.7 g, 46.2 mmol, 5.0 eq) 1.0 hr later. The mixture was stirred at 26°C for 16 h and concentrated under reduced pressure. The residue was purified by acidic preparative HPLC to give tert-butyl trans-3-hydroxy-4-(((4-methoxybenzyl)(methyl)amino)methyl)pyrrolidine-1-carboxylate (3.0 g, 8.5 mmol, 92.5% yield) as a colorless oil. ESI [M+H] = 351.1

[0760]

[0761] tert-Butyl trans-3-hydroxy-4-((methylamino)methyl)pyrrolidine-1-carboxylate, M25. To a solution of tert-butyl trans-3-hydroxy-4-(((4-methoxybenzyl)(methyl)amino)methyl)pyrrolidine-1-carboxylate (1.8 g, 5.1 mmol, 1.0 eq) in MeOH (25 mL) was added Pd(OH)2 (72.1 mg, 513 umol, 0.1 eq.) and the mixture was stirred at 26°C under 50 psi H2 for 16 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure to give tert-butyl trans-3-hydroxy-4-((methylamino)methyl)pyrrolidine-1-carboxylate (1.1 g, 4.7 mmol, 92.9% yield) as a yellow oil, which was used directly without any purification. ESI [M+H] = 231.1

[0762]

[0763]

[0146] tert-Butyl trans-3-((((benzyloxy)carbonyl)(methyl)amino)methyl)-4-hydroxypyrrolidine-1-carboxylate, M26. To a solution of tert-butyl trans-3-hydroxy-4-((methylamino)methyl)pyrrolidine-1-carboxylate (1.1 g, 4.7 mmol, 1.0 eq) in THF (15 mL) and H2O (15 mL) was added K2CO3 (1.3 g, 9.5 mmol, 2.0 eq) and CbzCl (1.2 g, 7.1 mmol, 1.5 eq.) at 0°C. The mixture was stirred at 26°C for 16 hr and concentrated under reduced pressure. The residue was purified by basic preparative HPLC to yield tert-butyl trans-3-((((benzyloxy)carbonyl)(methyl)amino)methyl)-4-hydroxypyrrolidine-1-carboxylate (800 mg, 2.2 mmol, 45.9% yield) as a yellow oil. ESI[M+H]=365.1

[0764]

[0765] Benzyl ((trans-4-hydroxypyrrolidin-3-yl)methyl)(methyl)carbamate, M27. tert-Butyl trans-3-((((benzyloxy)carbonyl)(methyl)amino)methyl)-4-hydroxypyrrolidine-1-carboxylate (400 mg, 1.1 mmol, 1.0 eq) was dissolved in HCl / MeOH (4 M, 10 mL) and the mixture was stirred at 26° C. for 1 hour. The mixture was concentrated under reduced pressure and then dissolved in MeOH (15 mL). The pH was adjusted to 8-9 by basic resin and the mixture was filtered. The filtrate was concentrated to give benzyl ((trans-4-hydroxypyrrolidin-3-yl)methyl)(methyl)carbamate (120 mg, 454 umol, 41.2% yield) as a yellow oil. [M+H]=264.9

[0766]

[0767] General Procedures

[0768]

[0769] (S)-tert-Butyl 3-(4-hydroxypiperidine-1-carbonyl)pyrrolidine-1-carboxylate, M29. To a solution of (3S)-1-tert-butoxycarbonylpyrrolidine-3-carboxylic acid (1.0 g, 4.6 mmol, 1.0 eq.) and piperidin-4-ol (1.4 g, 13.9 mmol, 3.0 eq) in DCM (20 mL) was added HATU (2.3 g, 6.0 mmol, 1.3 eq) and DIEA (1.5 g, 11.6 mmol, 2.0 mL, 2.5 eq). The mixture was stirred at 20 °C for 1 hour and diluted with DCM (100 mL). The organic layer was washed with brine (20 mL), dried over anhydrous Na 2 SO 4 , filtered and concentrated in vacuo to give (S)-tert-butyl 3-(4-hydroxypiperidine-1-carbonyl)pyrrolidine-1-carboxylate (1.2 g, crude) as a yellow oil, which was used in the next step without further purification. 1 HNMR (400 MHz, chloroform-d) δ = 3.69-3.62 (m, 1H), 3.48-3.31 (m, 1H), 3.20 (br.s., 1H), 2.90 (d, J = 7.1 Hz, 1H), 2.71 (d, J = 5.3 Hz, 2H), 2.28 (br.s., 2H), 2.13 (br.s., 2H), 1.92-1.79 (m, 3H), 1.52 (d, J = 8.8 Hz, 3H), 1.46-1.35 (m, 9H). ESI [M+H] = 299.0

[0770]

[0771] (R)-tert-Butyl 3-((4-hydroxypiperidin-1-yl)methyl)pyrrolidine-1-carboxylate, M30. To a solution of (S)-tert-butyl 3-(4-hydroxypiperidin-1-carbonyl)pyrrolidine-1-carboxylate (1.2 g, 4.0 mmol, 1.0 eq.) in THF (30 mL) was added LiAlH4 (183 mg, 4.8 mmol, 1.2 eq.) at 0°C. The mixture was stirred at 20°C for 1 hour, quenched with saturated aqueous MgSO4 (20 mL), and filtered. The filtrate was concentrated and purified by silica gel chromatography (DCM:MeOH = 10:1) to give tert-butyl (R)-3-((4-hydroxypiperidin-1-yl)methyl)pyrrolidine-1-carboxylate (600 mg, crude) as a yellow oil. ESI [M+H] = 285.2

[0772]

[0773] (S)-1-(Pyrrolidin-3-ylmethyl)piperidin-4-ol, M31. A solution of (R)-tert-butyl 3-((4-hydroxypiperidin-1-yl)methyl)pyrrolidine-1-carboxylate (600 mg, 2.1 mmol, 1.0 eq.) in HCl / MeOH (4 M, 10 mL) was stirred at 20° C. for 0.5 h and then concentrated under reduced pressure. The residue was diluted with MeOH (20 mL), basified to pH 7-8 by passing through a basic resin and filtered. The filtrate was concentrated to give (S)-1-(pyrrolidin-3-ylmethyl)piperidin-4-ol (400 mg, crude) as a light yellow oil, which was used in the next step without further purification. ESI [M+H] = 184.9

[0774]

[0775] General Procedure Q

[0776]

[0777] Benzyl 3-((3-((tert-Butoxycarbonyl)amino)propyl)carbamoyl)pyrrolidine-1-carboxylate, M33. To a mixture of 1-benzyloxycarbonylpyrrolidine-3-carboxylic acid (300 mg, 1.2 mmol, 1.0 eq), DIEA (466 mg, 3.6 mmol, 3.0 eq) and HATU (686 mg, 1.8 mmol, 1.5 eq) in DCM (20 mL) was added tert-butyl N-(3-aminopropyl)carbamate (418 mg, 2.4 mmol, 2.0 eq) in one portion at 20° C. The mixture was stirred at 20° C. for 0.5 h and then poured into ice water (50 mL). The aqueous layer was extracted with ethyl acetate (20 mL*3). The combined organic phases were washed with brine (10 mL*2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by preparative TLC (SiO 2 , ethyl acetate) to give benzyl 3-((3-((tert-butoxycarbonyl)amino)propyl)carbamoyl)pyrrolidine-1-carboxylate (450 mg, 654 umol, 54.5% yield, 59% purity) as a colorless oil. ESI [M+H] = 406.1

[0778]

[0779] Benzyl 3-(((3-((tert-Butoxycarbonyl)amino)propyl)amino)methyl)pyrrolidine-1-carboxylate, M34. To a solution of benzyl 3-((3-((tert-Butoxycarbonyl)amino)propyl)carbamoyl)pyrrolidine-1-carboxylate (400 mg, 986 umol, 1.0 eq) in THF (5 mL) was added BH3.THF (1 M, 9.8 mL, 10 eq.) and the reaction mixture was warmed to 70 °C for 1 hr. The reaction was slowly quenched with methanol (100 mL) and concentrated under reduced pressure at 40 °C to give crude benzyl 3-[[3-(tert-Butoxycarbonylamino)propylamino]methyl]pyrrolidine-1-carboxylate (400 mg, crude) as a colorless oil. ESI [M+H] = 392.1

[0780]

[0781]

[0266] Benzyl 3-(((tert-butoxycarbonyl)(3-((tert-butoxycarbonyl)amino)propyl)amino)methyl)pyrrolidine-1-carboxylate, M35. To a solution of benzyl 3-[[3-(tert-butoxycarbonylamino)propylamino]methyl]pyrrolidine-1-carboxylate (400 mg, 1.0 mmol, 1.0 eq) and tert-butyl tert-butoxycarbonyl carbonate (1.3 g, 6.1 mmol, 6.0 eq) in MeOH (50 mL) was added TEA (309 mg, 3.0 mmol, 3.0 eq) in one portion and the mixture was stirred at 20 °C for 10 h. The mixture was concentrated and purified by preparative HPLC (neutral conditions) to give benzyl 3-(((tert-butoxycarbonyl)(3-((tert-butoxycarbonyl)amino)propyl)amino)methyl)pyrrolidine-1-carboxylate as a colorless oil (200 mg, 227 umol, 22.3% yield, 56% purity). ESI [M+H] = 492.4

[0782]

[0783] tert-Butyl (3-((tert-butoxycarbonyl)amino)propyl)(pyrrolidin-3-ylmethyl)carbamate, M36. To a solution of benzyl 3-(((tert-butoxycarbonyl)(3-((tert-butoxycarbonyl)amino)propyl)amino)methyl)pyrrolidine-1-carboxylate (100 mg, 203 umol, 1.0 eq.) in EtOAc (30 mL) was added Pd / C (100 mg, 203 umol, 1.0 eq) and the mixture was stirred at 20 °C under 50 psi H2 for 1 hr. The mixture was filtered and the filtrate was concentrated in vacuo to give tert-butyl (3-((tert-butoxycarbonyl)amino)propyl)(pyrrolidin-3-ylmethyl)carbamate (70 mg, crude) as a colorless oil. ESI [M+H] = 358.2

[0784] Amine synthesis

[0785]

[0786] (3R,4R)-4-(Dimethylamino)pyrrolidin-3-ol, M37. Synthesized using General Procedure K with (2R)-2-hydroxy-2-phenyl-acetic acid substituted for (2S)-2-hydroxy-2-phenyl-acetic acid. ESI [M+H] = 130.8

[0787]

[0788] (3S,4R)-4-(Dimethylamino)pyrrolidin-3-ol, M38. Synthesized using General Procedure L with (3R,4R)-3-amino-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester replacing (3S4S)-3-amino-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester. 1H NMR (400 MHz, Methanol-d4) δ = 4.60 (br. s., 1H), 3.81-3.63 (m, 2H), 3.48-3.37 (m, 3H), 2.82 (s, 6H). ESI [M+H] = 130.8

[0789]

[0790] (3S,4R)-4-((Dimethylamino)methyl)pyrrolidin-3-ol, M39. Synthesized using General Procedure M substituting tert-butyl (3S,4S)-3-((((benzyloxy)carbonyl)amino)methyl)-4-hydroxypyrrolidine-1-carboxylate for tert-butyl (3R,4R)-3-((((benzyloxy)carbonyl)amino)methyl)-4-hydroxypyrrolidine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 4.44 (t, J = 3.3Hz, 1H), 3.50 (dd, J = 8.6, 11.3Hz, 1H), 3.40-3.31 (m, 2H), 3. 27(s,1H),3.08(t,J=11.3Hz,1H),2.96(dd,J=6.8,12.7Hz,1H),2.83-2.75(m,1H),2.54(s,6H). ESI[M+H]=145.0

[0791]

[0792] (3R,4R)-4-((Dimethylamino)methyl)pyrrolidin-3-ol, M40. Synthesized using General Procedure N substituting tert-butyl (3S,4S)-3-((((benzyloxy)carbonyl)amino)methyl)-4-hydroxypyrrolidine-1-carboxylate for tert-butyl (3R,4R)-3-((((benzyloxy)carbonyl)amino)methyl)-4-hydroxypyrrolidine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 4.29-4.19 (m, 1H), 3.55 (dd, J = 6.8, 11.7Hz, 1H), 3.34 (dd, J = 4.9, 12.3Hz, 1H ), 3.15 (dd, J = 3.1, 12.3Hz, 1H), 3.06 (dd, J = 4.9, 11.9Hz, 1H), 2.54-2.39 (m, 3H), 2.39-2.28 (s, 6H). ESI[M+H]=145.0

[0793]

[0794] Benzyl (((3R,4R)-4-hydroxypyrrolidin-3-yl)methyl)(methyl)carbamate, M41.

[0795] Synthesized using General Procedure O with (3S,4R)-3-(aminomethyl)-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester replacing trans-3-(aminomethyl)-4-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester. 1H NMR (400 MHz, CHLOROFORM-d) δ = 7.38-7.26 (m, 5H), 5.16-5.04 (m, 2H), 4.16-3.91 (m, 1H), 3.35 (d, J = 7.5 Hz, 1H), 3.17 (br. s., 2H), 3.00-2.73 (m, 4H), 2.25 (br. s., 1H), 2.11 (br. s., 2H). ESI [M+H] = 265.1

[0796]

[0797] Benzyl (((3S,4S)-4-Hydroxypyrrolidin-3-yl)methyl)(methyl)carbamate, M42.

[0798] Synthesized using General Procedure O with (3R,4S)-3-(aminomethyl)-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester replacing trans-3-(aminomethyl)-4-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester. ESI [M+H] = 265.1

[0799]

[0800] Benzyl (((3R,4S)-4-hydroxypyrrolidin-3-yl)methyl)(methyl)carbamate, M43.

[0801] Synthesized using General Procedure O with (3S,4S)-3-(aminomethyl)-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester replacing trans-3-(aminomethyl)-4-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester. ESI [M+H] = 265.1

[0802]

[0803] (R)-2-(((R)-Pyrrolidin-3-ylmethyl)amino)propan-1-ol, M44. Synthesized using General Procedure P with (3R)-1-tert-butoxycarbonylpyrrolidine-3-carboxylic acid replacing (3S)-1-tert-butoxycarbonylpyrrolidine-3-carboxylic acid and (R)-2-aminopropan-1-ol replacing piperidin-4-ol. ESI [M+H] = 159.1

[0804]

[0805] (R)-2-(((S)-Pyrrolidin-3-ylmethyl)amino)propan-1-ol, M45. Synthesized using General Procedure P substituting (R)-2-aminopropan-1-ol for piperidin-4-ol. 1 H NMR (400MHz, methanol-d4) δ = 3.84 (dd, J = 3.7, 12.1Hz, 1H), 3.67-3.54 (m, 2H), 3.52-3.37 (m, 2H), 3.34 (s, 1H), 3.28-3.20 (m, 2H), 3.1 1(dd,J=9.0,11.7Hz,1H),2.85-2.73(m,1H),2.35(dd,J=4.6,13.0Hz,1H),1.85(qd,J=8.7,13.2Hz,1H),1.35(d,J=6.6Hz,3H). ESI[M+H]=159.1

[0806]

[0807] (S)-2-(((R)-Pyrrolidin-3-ylmethyl)amino)propan-1-ol, M46. Synthesized using General Procedure P, substituting (R)-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid for (S)-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid and (S)-2-aminopropan-1-ol for piperidin-4-ol. ESI [M+H] = 159.1

[0808]

[0809] (S)-2-(((S)-Pyrrolidin-3-ylmethyl)amino)propan-1-ol, M47. Synthesized using General Procedure P with (S)-2-aminopropan-1-ol replacing piperidin-4-ol. ESI [M+H] = 159.1

[0810]

[0811] (R)-3-Methyl-2-(((R)-pyrrolidin-3-ylmethyl)amino)butan-1-ol, M48. Synthesized using General Procedure P with (R)-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid replacing (S)-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid and (R)-2-amino-3-methylbutan-1-ol replacing piperidin-4-ol. ESI [M+H] = 187.1

[0812]

[0813] (R)-3-Methyl-2-(((S)-pyrrolidin-3-ylmethyl)amino)butan-1-ol, M49. Synthesized using General Procedure P with (R)-2-amino-3-methylbutan-1-ol replacing piperidin-4-ol. ESI [M+H] = 187.1

[0814]

[0815] (S)-3-Methyl-2-(((R)-pyrrolidin-3-ylmethyl)amino)butan-1-ol, M50. Synthesized using General Procedure P, substituting (R)-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid for (S)-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid and (S)-2-amino-3-methylbutan-1-ol for piperidin-4-ol. ESI [M+H] = 187.1

[0816]

[0817] (S)-3-Methyl-2-(((S)-pyrrolidin-3-ylmethyl)amino)butan-1-ol, M51. Synthesized using General Procedure P with (S)-2-amino-3-methylbutan-1-ol replacing piperidin-4-ol. ESI [M+H] = 187.1

[0818]

[0819] (1R,4r)-4-(((R)-pyrrolidin-3-ylmethyl)amino)cyclohexanol, M52. Synthesized using General Procedure P substituting (R)-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid for (S)-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid and (1r,4r)-4-aminocyclohexanol for piperidin-4-ol. 1H NMR (400MHz, methanol-d4) δ=3.52-3.46(m,2H),3.43-3.42(m,1H),3.32-3.22(m,1H),2.96-2.90(m,3H),2.88-2 .86(m,1H),2.54-2.52(m,1H),2.02-2.01(m,1H),1.98-1.96(m,4H),1.73-1.71(m,1H),1.31-1.26(m,4H). ESI[M+H]=199.1

[0820]

[0821] (1S,4r)-4-(((S)-Pyrrolidin-3-ylmethyl)amino)cyclohexanol, M53. Synthesized using General Procedure P with (1r,4r)-4-aminocyclohexanol replacing piperidin-4-ol. 1H NMR (400 MHz, Methanol-d4) δ = 3.56-3.51 (m, 2H), 3.07-3.01 (m, 4H), 2.66-2.62 (m, 1H), 2.31-2.28 (m, 1H), 2.13-2.02 (m, 4H), 1.80-1.77 (m, 1H), 1.44-1.28 (m, 6H). ESI [M+H] = 199.1

[0822]

[0823] (R)-2-Methyl-2-((pyrrolidin-3-ylmethyl)amino)propan-1-ol, M54. Synthesized using General Procedure P, substituting (R)-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid for (S)-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid and 2-amino-2-methylpropan-1-ol for piperidin-4-ol. 1H NMR (400 MHz, Methanol-d4) δ = 3.57-3.53 (m, 3H), 3.41-3.38 (m, 1H), 3.09-3.02 (m, 3H), 2.93 (s, 1H), 2.67-2.64 (m, 1H), 2.32-2.31 (m, 1H), 1.82-.177 (m, 1H), 1.30 (s, 6H). ESI[M+H]=173.2

[0824]

[0825] (S)-2-Methyl-2-((pyrrolidin-3-ylmethyl)amino)propan-1-ol, M55. Synthesized using General Procedure P with 2-amino-2-methylpropan-1-ol replacing piperidin-4-ol. 1H NMR (400 MHz, Methanol-d4) δ = 3.46-3.41 (m, 3H), 3.33-3.30 (m, 1H), 3.22 (m, 1H), 2.97-2.96 (m, 1H), 2.88-2.86 (m, 2H), 2.54-2.50 (m, 1H), 2.25-2.23 (m, 1H), 1.76-1.71 (m, 1H), 1.19 (s, 6H). ESI [M+H] = 173.2

[0826]

[0827] (R)-1-(Pyrrolidin-3-ylmethyl)piperidin-4-ol, M56. Synthesized using General Procedure P with (R)-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid replacing (S)-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid. ESI [M+H] = 185.1

[0828]

[0829] tert-Butyl (3-((tert-Butoxycarbonyl)(methyl)amino)propyl)(pyrrolidin-3-ylmethyl)carbamate, M57. Synthesized using General Procedure Q substituting tert-butyl (3-aminopropyl)(methyl)carbamate for tert-butyl N-(3-aminopropyl)carbamate. ESI [M+H] = 372.2

[0830]

[0831] Tert-butyl (3-(dimethylamino)propyl)(pyrrolidin-3-ylmethyl)carbamate, M58.

[0832] Synthesized using General Procedure Q with N,N-dimethylpropane-1,3-diamine replacing tert-butyl N-(3-aminopropyl)carbamate. ESI [M+H] = 286.1

[0833]

[0834] tert-Butyl 4-(pyrrolidin-3-ylmethyl)piperazine-1-carboxylate, M59. Synthesized using General Procedure Q with tert-butyl piperazine-1-carboxylate replacing tert-butyl N-(3-aminopropyl)carbamate. ESI [M+H] = 270.0

[0835]

[0836] tert-Butyl (2-((tert-Butoxycarbonyl)amino)ethyl)(pyrrolidin-3-ylmethyl)carbamate, M60. Synthesized using General Procedure Q substituting tert-butyl (2-aminoethyl)carbamate for tert-butyl N-(3-aminopropyl)carbamate. ESI [M+H] = 344.1

[0837]

[0838] tert-Butyl (2-((tert-Butoxycarbonyl)amino)ethyl)(pyrrolidin-3-ylmethyl)carbamate, M61. Synthesized using General Procedure Q substituting tert-butyl (2-aminoethyl)(methyl)carbamate for tert-butyl N-(3-aminopropyl)carbamate. ESI [M+H] = 358.1

[0839] tert-Butyl (2-(dimethylamino)ethyl)(pyrrolidin-3-ylmethyl)carbamate, M62.

[0840] Use the general procedure Q with N 1 ,N 1 -dimethylethane-1,2-diamine was used instead of tert-butyl N-(3-aminopropyl)carbamate. ESI [M+H] = 272.1

[0841]

[0842] tert-Butyl (2-((tert-Butoxycarbonyl)amino)propyl)(pyrrolidin-3-ylmethyl)carbamate, M63. Synthesized using General Procedure Q substituting tert-butyl (1-aminopropan-2-yl)carbamate for tert-butyl N-(3-aminopropyl)carbamate. ESI [M+H] = 358.1

[0843]

[0844] tert-Butyl (2-hydroxyethyl)(pyrrolidin-3-ylmethyl)carbamate, M64. Synthesized using General Procedure Q substituting 2-aminoethanol for tert-butyl N-(3-aminopropyl)carbamate. ESI [M+H] = 245.1

[0845]

[0846] tert-Butyl (3-hydroxypropyl)(pyrrolidin-3-ylmethyl)carbamate, M65. Synthesized using general procedure Q substituting 3-aminopropan-1-ol for tert-butyl N-(3-aminopropyl)carbamate. ESI [M+H] = 259.1

[0847]

[0848] 4-(7-(3-(Dimethylamino)piperidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 2. Synthesized using General Procedure A substituting N,N-dimethylpiperidin-3-amine for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 9.13 (br.s., 1H), 7.85-7.64 (m, 5H), 7.61 (d, J = 8.8Hz, 1H) ,7.33(d,J=2.3Hz,1H),7.28(br.s.,1H),7.24-7.18(m,1H),5.26(s,2H),3.90(d ,J=11.8Hz,1H),3.63(d,J=12.5Hz,1H),3.48(br.s.,1H),3.11(t,J=9.5Hz,2H), 3.01(d,J=3.8Hz,6H),2.18(br.s.,1H),2.01(d,J=9.3Hz,1H),1.93-1.80(m,2H). ESI[M+H]=450.2

[0849]

[0850] 4-(7-(trans-Piperolo[3,2-b]pyrrol-1(2H)-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 3. Synthesized using General Procedure A with trans-piperolo[3,2-b]pyrrole-1(2H)-carboxylic acid tert-butyl ester replacing piperazine-1-carboxylic acid tert-butyl ester. 1 H NMR (400 MHz, methanol-d4) δ = 9.06 (br. s., 1H), 7.77-7.70 (m, 2H), 7.70-7.61 (m, 3H), 7.54 (d, J = 8.8 Hz, 1H), 7.25 (s, 1H), 6.95 (d, J = 2.2 Hz, 1H), 6.89-6.83 (m, 1H), 5.22 (s, 2 H),4.34-4.19(m,1H),4.13-3.97(m,2H),3.90-3.76(m,2H),3.69(dt,J=5.5,11.1H z,1H),2.79-2.67(m,1H),2.39-2.25(m,1H),2.23-2.10(m,1H),2.08-1.97(m,1H). ESI[M+H]=434.2

[0851]

[0852] (R)-4-(7-(3-(methylamino)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 4. was synthesized using general procedure A substituting tert-butyl (R)-methyl(pyrrolidin-3-yl)carbamate for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 9.06 (s, 1H), 7.83-7.60 (m, 5H), 7.57 (d, J = 8.8Hz, 1H), 7.26 (d, J = 1.5Hz, 1H), 6.94 (d, J = 2.3Hz, 1H), 6.83 (dd, J = 2. 6,8.9Hz,1H),5.24(s,2H),4.03(br.s.,1H),3.77-3.60(m,3H),3.53-3.46(m,1H),2.83(s,3H),2.65-2.51(m,1H),2.30(d,J=5.3Hz,1H). ESI[M+H]=422.2

[0853]

[0854] (S)-4-(7-(3-(methylamino)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopentan-12-yl)benzonitrile, 5. was synthesized using general procedure A substituting tert-butyl (S)-methyl(pyrrolidin-3-yl)carbamate for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 9.06 (s, 1H), 7.83-7.60 (m, 5H), 7.57 (d, J = 8.8Hz, 1H), 7.26 (d, J = 1.5Hz, 1H), 6.94 (d, J = 2.3Hz, 1H), 6.83 (dd, J = 2. 6,8.9Hz,1H),5.24(s,2H),4.03(br.s.,1H),3.77-3.60(m,3H),3.53-3.46(m,1H),2.83(s,3H),2.65-2.51(m,1H),2.30(d,J=5.3Hz,1H). ESI[M+H]=422.2

[0855]

[0856] 4-(7-(3-((methylamino)methyl)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopentan-12-yl)benzonitrile, 18. Synthesized using general procedure A substituting tert-butyl methyl(pyrrolidin-3-ylmethyl)carbamate for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 9.17 (br.s., 1H), 7.73-7.60 (m, 5H), 7.47 (d, J = 7.5Hz, 1 H),7.26(br.s.,1H),6.78(br.s.,1H),6.68(d,J=7.5Hz,1H),5.14(br.s.,2H) ,3.59(br.s.,1H),3.55-3.46(m,1H),3.42(br.s.,1H),3.15(br.s.,3H),2.77 (s,3H),2.75-2.66(m,1H),2.32(d,J=6.2Hz,1H),1.88(dd,J=8.4,11.5Hz,1H). ESI[M+H]=436.2

[0857]

[0858] (S)-4-(7-(3-(Dimethylamino)piperidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 20. Synthesized using general procedure C substituting tert-butyl (S)-piperidin-3-ylcarbamate for tert-butyl (azetidin-3-ylmethyl)(methyl)carbamate. 1 H NMR (400MHz, methanol-d4) δ = 9.06 (br.s., 1H), 7.80-7.64 (m, 4H), 7.62 (s, 1H), 7.58 (d, J = 8.8H z,1H),7.31(d,J=2.6Hz,1H),7.25(s,1H),7.19(dd,J=2.6,8.8Hz,1H),5.23(s,2H),3.88 (d,J=11.9Hz,1H),3.66-3.56(m,1H),3.45(t,J=9.0Hz,1H),3.35(br.s.,1H),3.13-3.05 (m,1H),3.04-2.86(m,6H),2.24-2.12(m,1H),1.99(d,J=11.5Hz,1H),1.92-1.77(m,2H). ESI[M+H]=450.2

[0859]

[0860] 4-(7-(trans-3-(dimethylamino)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 57. Synthesized using general procedure A with trans-4-(dimethylamino)pyrrolidin-3-ol replacing tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ=9.07(s,1H),7.75-7.69(m,2H),7.68-7.60(m,3H),7.54(d,J=8.8Hz,1H),7.23(s,1H),6.89(d,J=2.2Hz,1H),6.79(dd ,J=2.4,8.6Hz,1H),5.20(s,2H),4.72(q,J=6.6Hz,1H),3.96-3.82(m,3H),3.59(dd,J=6.8,9.5Hz,1H),3.28-3.22(m,1H),3.14-2.99(m,6H). ESI[M+H]=452.2

[0861]

[0862] 4-(7-((3S,4S)-3-(Dimethylamino)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 58. Synthesized using General Procedure A substituting (3S,4S)-4-(dimethylamino)pyrrolidin-3-ol for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ=9.07(br.s.,1H),7.75-7.58(m,5H),7.53(d,J=8.8Hz,1H),7.22(br.s.,1H),6.88(br.s.,1H),6.78(d,J= 8.4Hz,1H),5.18(br.s.,2H),4.73(d,J=6.6Hz,1H),3.99-3.79(m,3H),3.60(t,J=7.7Hz,1H),3.27(br.s.,1H),3.08(br.s.,6H). ESI[M+H]=452.2

[0863]

[0864] 4-(7-(cis-3-hydroxy-4-((methylamino)methyl)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 73. Synthesized using general procedure A substituting cis-4-((dimethylamino)methyl)pyrrolidin-3-ol for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 9.02 (br.s., 1H), 7.77-7.60 (m, 5H), 7.50 (d, J = 8.8H z,1H),7.23(s,1H),6.79(d,J=2.2Hz,1H),6.70(dd,J=2.6,8.8Hz,1H),5.1 8(s,2H),4.57(br.s.,1H),3.72-3.56(m,2H),3.49-3.38(m,2H),3.23(dd, J=5.7,12.8Hz,1H),3.00(d,J=11.9Hz,1H),2.78(s,3H),2.71(br.s.,1H). ESI[M+H]=452.1

[0865]

[0866] 4-(7-(trans-3-((dimethylamino)methyl)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 74. Synthesized using general procedure A with trans-4-((dimethylamino)methyl)pyrrolidin-3-ol replacing tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ=9.15(br.s.,1H),7.67-7.51(m,5H),7.42(d,J=8.4Hz,1H),7.19(br.s.,1H),6.71(br.s.,1H),6.61(d,J=7.9Hz, 1H),5.10(br.s.,2H),4.28-4.15(m,1H),3.74-3.58(m,2H),3.38-3.25(m,2H),3.20-3.07(m,2H),2.97-2.86(m,6H),2.69-2.60(m,1H). ESI[M+H]=466.2

[0867]

[0868] 4-(7-(cis-3-((dimethylamino)methyl)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 75. Synthesized using general procedure A substituting cis-4-((dimethylamino)methyl)pyrrolidin-3-ol for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 9.10 (s, 1H), 7.79-7.59 (m, 5H), 7.49 (d, J = 8.8Hz, 1H), 7.24 (br.s., 1H), 6.79 (d, J = 1.8Hz, 1H), 6.69 (d, J = 8. 4Hz,1H),5.17(s,2H),4.57(br.s.,1H),3.74-3.53(m,4H),3.44(d,J=10.6Hz,1H),3.24(br.s.,1H),2.98(s,6H),2.86(br.s.,1H). ESI[M+H]=466.2

[0869]

[0870] 4-(7-(cis-3-amino-4-fluoropyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 76. Synthesized using general procedure A substituting tert-butyl (cis-4-fluoropyrrolidin-3-yl)carbamate for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 9.06 (br.s., 1H), 7.77-7.70 (m, 2H), 7.69-7.61 (m, 3H), 7.55 (d, J = 8.8Hz, 1H), 7.24 (s, 1H), 6.89 (d, J = 2.6Hz, 1H), 6.77 (dd, J=2.2,8.8Hz,1H),5.21(s,2H),4.25-4.11(m,1H),3.91(t,J=8.8Hz,1H), 3.87-3.81(m,1H),3.77(s,1H),3.51(t,J=9.3Hz,1H),3.41-3.32(m,1H). ESI[M+H]=426.1

[0871]

[0872] 4-(7-((3S,4S)-3-((dimethylamino)methyl)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopyridine-12-yl)benzonitrile, 81.

[0873] Synthesized using general procedure A substituting (3S,4R)-4-((dimethylamino)methyl)pyrrolidin-3-ol for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 9.02 (s, 1H), 7.74-7.60 (m, 5H), 7.48 (d, J = 8.6Hz, 1H), 7.22 (s, 1H), 6.78 (d, J = 2.3Hz, 1H), 6.69 (dd, J = 2.3, 8.6Hz, 1H), 5.16(s,2H),4.57(br.s.,1H),3.72-3.54(m,3H),3.44(d,J=10.2Hz,1H ),3.34(br.s.,1H),3.28-3.22(m,1H),2.99(s,6H),2.91-2.81(m,1H). ESI[M+H]=466.2

[0874]

[0875] 4-(7-((3R,4R)-3-((dimethylamino)methyl)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopyridine-12-yl)benzonitrile, 82.

[0876] Synthesized using general procedure A substituting (3R,4S)-4-((dimethylamino)methyl)pyrrolidin-3-ol for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ=9.01(br.s.,1H),7.74-7.60(m,5H),7.48(d,J=8.6Hz,1H),7.21(s,1H),6.78(d,J=2.3Hz,1H),6.69(dd,J=2.3,9.0Hz,1H ),5.16(s,2H),4.57(br.s.,1H),3.73-3.55(m,3H),3.44(d,J=10.6Hz,1 H),3.34(br.s.,1H),3.29-3.22(m,1H),2.99(s,6H),2.90-2.82(m,1H). ESI[M+H]=466.2

[0877]

[0878] 4-(7-((3R,4S)-3-(Dimethylamino)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 83. Synthesized using General Procedure A substituting (3S,4R)-4-(dimethylamino)pyrrolidin-3-ol for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 9.05 (s, 1H), 7.74-7.67 (m, 2H), 7.67-7.60 (m, 3H), 7.52 (d, J = 8.8Hz, 1H), 7.25 (br.s., 1H), 6.86 (d, J = 1.8Hz, 1H), 6 .74(d,J=7.1Hz,1H),5.19(s,2H),4.68(br.s.,1H),3.99-3.86(m,2H),3.69(dd,J=3.3,11.2Hz,1H),3.61-3.47(m,2H),3.13-2.90(m,6H). ESI[M+H]=452.2

[0879]

[0880] 4-(7-((R)-3-((((R)-1-hydroxypropan-2-yl)amino)methyl)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopentan-12-yl)benzonitrile, 85. Synthesized using General Procedure A with (R)-2-(((S)-pyrrolidin-3-ylmethyl)amino)propan-1-ol replacing tert-butyl piperazine-1-carboxylate. 1H NMR (400 MHz, methanol-d4) δ = 9.01 (s, 1H), 7.76-7.67 (m, 2H), 7.67-7.57 (m, 3H), 7.46 (d, J = 8.8 Hz, 1H), 7.21 (d, J = 1.3 Hz, 1H), 6.77 (d, J = 2.2 Hz, 1H), 6.68 (dd, J = 2.2, 8.8 Hz, 1H), 5.14 (s, 2H), 3.83 (dd, J = 3.7, 12.1Hz,1H),3.65-3.55(m,2H),3.51(dt,J=4.0,8.8Hz,1H),3.45-3.35(m,2H),3.23-3.14(m,3H),2 .73(td,J=7.3,14.9Hz,1H),2.39-2.29(m,1H),1.90(qd,J=8.2,12.3Hz,1H),1.33(d,J=7.1Hz,3H). ESI[M+H]=480.2

[0881]

[0882] 4-(7-((S)-3-((((S)-1-hydroxypropyl-2-yl)amino)methyl)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopyridine-12-yl)benzonitrile, 86.

[0883] Synthesized using General Procedure A with (S)-2-(((R)-pyrrolidin-3-ylmethyl)amino)propan-1-ol replacing tert-butyl piperazine-1-carboxylate. 1H NMR (400 MHz, Methanol-d4) δ = 9.04 (br. s., 1H), 7.75-7.57 (m, 5H), 7.47 (d, J = 8.8 Hz, 1H), 7.22 (br. s., 1H), 6.78 (d, J = 2.2 Hz, 1H), 6.68 (dd, J = 2.2, 8.8 Hz, 1H), 5.15 (s, 2H), 3.83 (dd, J = 3.5, 11.9 Hz z,1H),3.59(dd,J=5.7,11.9Hz,2H),3.55-3.47(m,1H),3.41(d,J=8.4Hz,2H),3.23-3.13(m,3 H), 2.80-2.66 (m, 1H), 2.35 (dd, J = 4.6, 11.7Hz, 1H), 1.96-1.85 (m, 1H), 1.33 (d, J = 6.6Hz, 3H). ESI[M+H]=480.2

[0884]

[0885] 4-(7-((3R,4S)-3-((dimethylamino)methyl)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopyridine-12-yl)benzonitrile, 108.

[0886] Synthesized using general procedure A substituting (3S,4S)-4-((dimethylamino)methyl)pyrrolidin-3-ol for tert-butyl piperazine-1-carboxylate. 1H NMR (400 MHz, methanol-d4) δ = 8.97 (s, 1H), 7.67-7.73 (m, 2H), 7.57-7.67 (m, 3H), 7.44 (d, J = 8.4 Hz, 1H), 7.18 (d, J = 1.3 Hz, 1H), 6.77 (d, J = 2.2 Hz, 1H), 6.66 (dd, J = 8.8, 2.2 Hz, 1 H),5.13(s,2H),4.26(q,J=6.0Hz,1H),3.62-3.76(m,2H),3.20(ddd,J=16.3,9.9,6 .4Hz,2H),2.98-3.07(m,1H),2.86-2.96(m,1H),2.70(s,6H),2.54-2.63ppm(m,1H). ESI[M+H]=466.1

[0887]

[0888] 4-(7-((3S,4R)-3-((dimethylamino)methyl)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopyridine-12-yl)benzonitrile, 109.

[0889] Synthesized using general procedure A substituting (3R,4R)-4-((dimethylamino)methyl)pyrrolidin-3-ol for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 8.97 (s, 1H), 7.74-7.68 (m, 2H), 7.67-7.57 (m, 3H), 7.45 (d,J=8.8Hz,1H),7.18(d,J=1.3Hz,1H),6.77(d,J=2.2Hz,1H),6.66(dd,J=2.4 ,8.6Hz,1H),5.13(s,2H),4.25(q,J=6.2Hz,1H),3.74-3.62(m,2H),3.27-3.14 (m,2H),3.08-2.95(m,1H),2.94-2.83(m,1H),2.68(s,6H),2.63-2.53(m,1H). ESI[M+H]=466.1

[0890]

[0891] (S)-4-(7-(3-((4-hydroxypiperidin-1-yl)methyl)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 110. Synthesized using General Procedure A with (R)-1-(pyrrolidin-3-ylmethyl)piperidin-4-ol replacing tert-butyl piperazine-1-carboxylate. 1H NMR (400 MHz, methanol-d4) δ = 9.00 (s, 1H), 7.78-7.60 (m, 5H), 7.49 (d, J = 8.6 Hz, 1H), 7.22 (s, 1H), 6.81 (d, J = 2.2 Hz, 1H), 6.74-6.68 (m, 1H), 5.18 (s, 2H), 4.11 (br. s., 1H), 3.85 (br. s.,1H),3.74-3.61(m,2H),3.57-3.41(m,3H),3.17-3.06(m,3H),2.88(d,J=7.9Hz,1H ), 2.36 (br.s., 1H), 2.17 (d, J = 13.9Hz, 1H), 2.08-1.86 (m, 4H), 1.77 (d, J = 13.7Hz, 1H). ESI[M+H]=506.2

[0892]

[0893] (R)-4-(7-(3-((4-hydroxypiperidin-1-yl)methyl)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 111. Synthesized using General Procedure A with (S)-1-(pyrrolidin-3-ylmethyl)piperidin-4-ol replacing tert-butyl piperazine-1-carboxylate. 1H NMR (400 MHz, methanol-d4) δ = 9.10 (br.s., 1H), 7.73-7.61 (m, 5H), 7.48 (d, J = 8.8 Hz, 1H), 7.28-7.21 (m, 1H), 6.78 (d, J = 2.2 Hz, 1H), 6.69 (dd, J = 2.6, 8.8 Hz, 1H), 5.17 (s, 2H), 4.10 (br.s .,1H),3.90-3.76(m,1H),3.65(t,J=8.2Hz,2H),3.56-3.34(m,4H),3.19-3.03(m,2H),2 .85(td,J=7.6,14.8Hz,1H),2.34(br.s.,1H),2.15(d,J=13.7Hz,1H),2.07-1.71(m,5H). ESI[M+H]=506.2

[0894]

[0895] 4-(7-(3-(Dimethylamino)piperidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 112. Synthesized using General Procedure C substituting 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl piperidin-3-ylcarbamate for tert-butyl (azetidin-3-ylmethyl)(methyl)carbamate. 1H NMR (400MHz, methanol-d4) δ = 8.07 (br.s., 1H), 7.84-7.67 (m, 6H), 7.63 (d, J = 8.8Hz, 1 H),7.38-7.30(m,2H),7.23(dd,J=2.9,9.0Hz,1H),5.36(s,2H),3.99(d,J=12. 3Hz,1H),3.70(s,1H),3.50-3.40(m,1H),3.25(s,1H),3.06(d,J=10.1Hz,1H), 2.98(s,6H),2.18(br.s.,1H),1.99(dd,J=4.4,8.8Hz,1H),1.90-1.79(m,2H). ESI[M+H]=449.2

[0896]

[0897] 4-(7-(trans-Piperolo[3,2-b]pyrrol-1(2H)-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 113. Synthesized using General Procedure A with 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl trans-piperillolo[3,2-b]pyrrole-1(2H)-carboxylate replacing tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 8.05 (br.s., 1H), 7.89-7.69 (m, 6H), 7.61 (d, J = 8.5Hz, 1H), 7.34 (br. s.,1H),6.99(br.s.,1H),6.96-6.88(m,1H),5.36(s,2H),4.68-4.49(m,1H),4.30(d,J=7.5H z,1H),4.14-4.00(m,1H),3.92-3.82(m,1H),3.74(d,J=5.3Hz,1H),3.45(d,J=7.8Hz,1H),2 .77(d,J=5.5Hz,1H),2.37(dd,J=6.1,10.7Hz,1H),2.22(d,J=10.5Hz,1H),2.06(br.s.,1H). ESI[M+H]=433.2

[0898]

[0899] (R)-4-(7-(3-(Methylamino)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 114. Synthesized using General Procedure A substituting 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl (R)-methyl(pyrrolidin-3-yl)carbamate for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 8.06 (s, 1H), 7.84-7.71 (m, 6H), 7.62 (d, J = 8.8Hz, 1H), 7.34 (s, 1H), 6.95 (d, J = 2.5Hz, 1H), 6.91-6.85 (m, 1H), 5.36 ( s,2H),4.10-4.00(m,1H),3.82-3.58(m,3H),3.51(d,J=6.0Hz,1H),2.83(s,3H),2.62-2.53(m,1H),2.32(d,J=5.8Hz,1H)ESI[M+H]=421.2

[0900]

[0901] (S)-4-(7-(3-(Methylamino)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 115. Synthesized using general procedure A substituting 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl (S)-methyl(pyrrolidin-3-yl)carbamate for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 8.06 (s, 1H), 7.84-7.71 (m, 6H), 7.62 (d, J = 8.8Hz, 1H), 7.34 (s, 1H), 6.95 (d, J = 2.5Hz, 1H), 6.91-6.85 (m, 1H), 5.36 ( s,2H),4.10-4.00(m,1H),3.82-3.58(m,3H),3.51(d,J=6.0Hz,1H),2.83(s,3H),2.62-2.53(m,1H),2.32(d,J=5.8Hz,1H)ESI[M+H]=421.2

[0902]

[0903] 4-(7-(3-(Aminomethyl)azetidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 118. Synthesized using general procedure A substituting 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl (azetidin-3-ylmethyl)carbamate for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 8.01 (br.s., 1H), 7.85-7.63 (m, 6H), 7.55 (d, J = 8.4Hz, 1H), 7.32 (br.s., 1H), 6.71 (br.s., 1H), 6.64(d,J=8.4Hz,1H),5.29(br.s.,2H),4.14(t,J=7.5Hz,2H),3.86-3.74(m,2H),3.31-3.26(m,2H),3.07(br.s.,1H). ESI[M+H]=407.1

[0904]

[0905] 4-(7-(3-((methylamino)methyl)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 122. Synthesized using general procedure A substituting 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl methyl(pyrrolidin-3-ylmethyl)carbamate for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 8.01 (br.s., 1H), 7.84-7.66 (m, 6H), 7.55 (d, J = 8.8H z,1H),7.31(s,1H),6.82(br.s.,1H),6.76(d,J=8.8Hz,1H),5.30(s,2H),3. 64(br.s.,1H),3.55(br.s.,1H),3.44(d,J=7.9Hz,1H),3.18(d,J=7.1Hz,3 H), 2.78 (s, 3H), 2.75 (br.s., 1H), 2.35 (d, J = 6.2Hz, 1H), 1.98-1.83 (m, 1H). ESI[M+H]=435.2

[0906]

[0907] (S)-4-(7-(3-(Dimethylamino)piperidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 123. Synthesized using General Procedure C substituting 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl (S)-piperidin-3-ylcarbamate (azetidin-3-ylmethyl)(methyl)carbamate. 1H NMR (400MHz, methanol-d4) δ = 8.06 (s, 1H), 7.85-7.68 (m, 6H), 7.63 (d, J = 8.8Hz, 1H), 7.38-7.29(m,2H),7.23(dd,J=2.6,8.8Hz,1H),5.35(s,2H),3.99(d,J=11.9H z,1H),3.76-3.63(m,1H),3.45(d,J=3.1Hz,1H),3.25(br.s.,1H),3.13-3.03 (m,1H),2.98(s,6H),2.27-2.13(m,1H),2.06-1.93(m,1H),1.89-1.78(m,2H). ESI[M+H]=449.2

[0908]

[0909] 4-(7-(3-Aminopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 124. Synthesized using general procedure A substituting 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl pyrrolidin-3-ylcarbamate for tert-butyl piperazine-1-carboxylate. 1HNMR (400MHz, methanol-d4) δ = 8.02 (d, J = 1.8Hz, 1H), 7.85-7.66 (m, 6H), 7.58 (d, J = 9.3Hz, 1H), 7.32 (s, 1H), 6.90 (d, J = 2.6Hz, 1H), 6.83 (dd, J = 2.6 ,8.8Hz,1H),5.33(s,2H),4.09(br.s.,1H),3.79-3.61(m,2H),3.52(dd,J=3.7,10.8Hz,2H),2.58-2.43(m,1H),2.24(dd,J=4.6,8.6Hz,1H). ESI[M+H]=407.1

[0910]

[0911] 4-(7-((3S,4S)-3-((dimethylamino)methyl)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 128. General Procedure A was used with 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1 The obtained product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazol-12-yl)benzonitrile with (3S,4R)-4-((dimethylamino)methyl)pyrrolidin-3-ol instead of tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 7.98 (d, J = 2.2Hz, 1H), 7.80-7.64 (m, 6H), 7.53 (d, J = 9.3Hz, 1H), 7.28 (d, J = 1.3Hz, 1H), 6.80 (d, J = 2.6Hz, 1H), 6.74(dd,J=2.6,8.8Hz,1H),5.29(s,2H),4.57(br.s.,1H),3.70-3.55(m,4H),3.44(d,J=10.6Hz,2H),2.97(s,6H),2.86(br.s.,1H). ESI[M+H]=465.1

[0912]

[0913] 4-(7-((3R,4R)-3-((dimethylamino)methyl)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 129. General Procedure A was used with 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1 The obtained product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazol-12-yl)benzonitrile with (3R,4S)-4-((dimethylamino)methyl)pyrrolidin-3-ol instead of tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 7.98 (s, 1H), 7.79-7.66 (m, 6H), 7.53 (d, J = 8.8Hz, 1H), 7.29 (s, 1H), 6.80 (d, J = 2.2Hz, 1H), 6.74 (dd, J=2.6,9.3Hz,1H),5.29(s,2H),4.56(br.s.,1H),3.71-3.53(m,4H),3.44(d,J=10.6Hz,2H),2.97(s,6H),2.86(br.s.,1H). ESI[M+H]=465.1

[0914]

[0915] 4-(7-((3R,4R)-3-(Dimethylamino)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 130. Synthesized using general procedure A substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (3R,4R)-4-(dimethylamino)pyrrolidin-3-ol for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 8.01 (d, J = 2.0Hz, 1H), 7.81-7.67 (m, 6H), 7.59 (d, J = 9.0Hz, 1H), 7.30 (d, J = 1.6Hz, 1H), 6.92 (d, J = 2.3Hz, 1H), 6.85 (dd ,J=2.5,9.0Hz,1H),5.32(s,2H),4.72(q,J=6.8Hz,1H),4.00-3.82(m,3H),3.60(dd,J=7.2,10.2Hz,1H),3.33(br.s.,1H),3.15-2.96(m,6H). ESI[M+H]=451.1

[0916]

[0917] 4-(7-((3S,4S)-3-(Dimethylamino)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 131. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (3S,4S)-4-(dimethylamino)pyrrolidin-3-ol for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ=7.99(d,J=1.8Hz,1H),7.79-7.66(m,6H),7.59(d,J=9.0Hz,1H),7.29(d,J=1.6Hz,1H),6.92(d,J=2.5Hz,1H),6.84( dd,J=2.5,9.0Hz,1H),5.31(s,2H),4.72(q,J=7.0Hz,1H),4.00-3.82(m,3H),3.60(dd,J=7.1,10.1Hz,1H),3.33(br.s.,1H),3.06(s,6H). ESI[M+H]=451.1

[0918]

[0919] 4-(7-((3S,4R)-3-(Dimethylamino)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 133. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (3R,4S)-4-(dimethylamino)pyrrolidin-3-ol for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ=7.99(br.s.,1H),7.79-7.66(m,6H),7.58(d,J=7.6Hz,1H),7.29(br.s.,1H),6.88(s,1H),6.82(d,J =8.8Hz,1H),5.31(s,2H),4.70(br.s.,1H),3.95(br.s.,2H),3.77-3.69(m,1H),3.63-3.52(m,2H),3.03(d,J=16.0Hz,6H). ESI[M+H]=451.1

[0920]

[0921] 4-(7-((3R,4S)-3-(dimethylamino)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepine-12-yl)benzonitrile, 134.

[0922] Synthesized using general procedure A with 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile and (3S,4R)-4-(dimethylamino)pyrrolidin-3-ol replacing tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ=8.02(br.s.,1H),7.82-7.67(m,6H),7.59(d,J=8.8Hz,1H),7.31(br.s.,1H),6.88(s,1H),6.82(d,J=8 .8Hz,1H),5.32(s,2H),4.70(br.s.,1H),3.95(br.s.,2H),3.73(dd,J=3.1,11.0Hz,1H),3.63-3.51(m,2H),3.03(br.s.,6H). ESI[M+H]=451.1

[0923]

[0924] 4-(7-((S)-3-((((R)-1-hydroxypropyl-2-yl)amino)methyl)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 135. The general procedure A was used with 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1 The reaction mixture was synthesized by using (R)-2-(((R)-pyrrolidin-3-ylmethyl)amino)propan-1-ol instead of tert-butyl piperazine-1-carboxylate and replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile. 1H NMR (400MHz, methanol-d4) δ = 7.98 (d, J = 2.0Hz, 1H), 7.81-7.68 (m, 6H), 7.54 (d, J = 9.0Hz, 1H) ,7.29(d,J=1.6Hz,1H),6.82(d,J=2.3Hz,1H),6.76(dd,J=2.5,9.0Hz,1H),5.29(s,2H) ,3.84(dd,J=3.8,12.0Hz,1H),3.70-3.51(m,3H),3.50-3.37(m,2H),3.26-3.14(m,3H ), 2.81-2.64 (m, 1H), 2.36 (s, 1H), 1.92 (dd, J = 8.6, 12.3Hz, 1H), 1.35 (d, J = 6.8Hz, 3H). ESI[M+H]=479.2

[0925]

[0926] 4-(7-((R)-3-((((R)-1-hydroxypropyl-2-yl)amino)methyl)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 136. General Procedure A was used with 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1 The synthesis was carried out by using (R)-2-(((S)-pyrrolidin-3-ylmethyl)amino)propan-1-ol instead of tert-butyl piperazine-1-carboxylate and replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile. 1 H NMR (400MHz, methanol-d4) δ = 7.96 (br.s., 1H), 7.77-7.65 (m, 6H), 7.52 (d, J = 8.8 Hz,1H),7.27(br.s.,1H),6.84-6.71(m,2H),5.27(s,2H),3.83(dd,J=4.0, 11.9Hz,1H),3.69-3.50(m,3H),3.47-3.36(m,2H),3.23-3.14(m,3H),2.7 9-2.69(m,1H),2.36(br.s.,1H),1.99-1.84(m,1H),1.33(d,J=6.6Hz,3H). ESI[M+H]=479.2

[0927]

[0928] 4-(7-((3S,4R)-3-((dimethylamino)methyl)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 137. General Procedure A was used with 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1 The synthesis was carried out by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazol-12-yl)benzonitrile with (3R,4R)-4-((dimethylamino)methyl)pyrrolidin-3-ol instead of tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ=7.99(br.s.,1H),7.79-7.66(m,6H),7.55(d,J=9.0Hz,1H),7.30(br.s.,1H),6.82(d,J=2.3Hz,1H),6.75(dd,J=2.3 ,8.6Hz,1H),5.29(s,2H),4.30(q,J=6.7Hz,1H),3.82-3.71(m,2H),3.46-3.34(m,2H),3.27-3.18(m,2H),2.99(s,6H),2.78-2.65(m,1H). ESI[M+H]=465.1

[0929]

[0930] 4-(7-((3R,4S)-3-((dimethylamino)methyl)-4-hydroxypyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 138. General Procedure A was used with 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1 The synthesis was carried out by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazol-12-yl)benzonitrile with (3S,4S)-4-((dimethylamino)methyl)pyrrolidin-3-ol instead of tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ=8.00(br.s.,1H),7.83-7.66(m,6H),7.54(d,J=8.8Hz,1H),7.30(br.s.,1H),6.81(d,J=2.4Hz,1H),6.74(dd,J=2.4,8 .8Hz,1H),5.29(s,2H),4.29(q,J=6.6Hz,1H),3.80-3.69(m,2H),3.44 -3.33(m,2H),3.26-3.19(m,2H),3.02-2.90(m,6H),2.75-2.61(m,1H). ESI[M+H]=465.2

[0931]

[0932] (S)-4-(7-(3-(((1-hydroxy-2-methylpropan-2-yl)amino)methyl)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 141. The general procedure A was used with 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1 The synthesis was carried out by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile with (R)-2-methyl-2-((pyrrolidin-3-ylmethyl)amino)propan-1-ol instead of tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 7.96 (br.s., 1H), 7.72 (dd, J = 7.4, 19.1Hz, 6H), 7.53 (d, J = 8. 8Hz,1H),7.27(br.s.,1H),6.81(br.s.,1H),6.76(d,J=9.0Hz,1H),5.28(s,2H),3. 72-3.61(m,1H),3.57(s,3H),3.50-3.41(m,1H),3.26-3.18(m,1H),3.16-3.08(m,2 H), 2.76-2.64 (m, 1H), 2.38 (d, J = 6.6Hz, 1H), 1.99-1.89 (m, 1H), 1.40-1.27 (m, 6H). ESI[M+H]=493.3

[0933]

[0934] (S)-4-(7-(3-((4-hydroxypiperidin-1-yl)methyl)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 142. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (R)-1-(pyrrolidin-3-ylmethyl)piperidin-4-ol for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 8.02 (br.s., 1H), 7.83-7.66 (m, 6H), 7.54 (d, J = 8.8Hz, 1H), 7. 31(br.s.,1H),6.82(s,1H),6.76(d,J=8.8Hz,1H),5.30(s,2H),4.11(br.s.,1H),3.8 4(br.s.,1H),3.69(t,J=8.2Hz,2H),3.61-3.37(m,4H),3.18(br.s.,1H),3.09(br.s .,1H),2.89(d,J=6.2Hz,1H),2.36(br.s.,1H),2.21-1.99(m,2H),1.99-1.67(m,4H). ESI[M+H]=505.3

[0935]

[0936] (R)-4-(7-(3-((4-hydroxypiperidin-1-yl)methyl)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 143. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (S)-1-(pyrrolidin-3-ylmethyl)piperidin-4-ol for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 8.01 (d, J = 2.2Hz, 1H), 7.78 (d, J = 1.8Hz, 1H), 7.76-7.67 (m, 5H), 7.53 (d, J = 8.8Hz, 1H) ,7.30(d,J=1.8Hz,1H),6.81(d,J=2.6Hz,1H),6.75(dd,J=2.6,8.8Hz,1H),5.29(s,2H),4.10(br.s.,1H),3.7 3-3.62(m,2H),3.58-3.49(m,1H),3.48-3.38(m,2H),3.32(br.s.,3H),3.22-3.01(m,2H),2.94-2.79(m,1H) ,2.35(br.s.,1H),2.15(d,J=11.0Hz,1H),2.03(d,J=11.5Hz,1H),1.97-1.84(m,2H),1.77(d,J=11.0Hz,1H). ESI[M+H]=505.2

[0937]

[0938] 4-(7-(2,6-diazaspiro[3.4]octan-6-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile, 146. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile and tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ=7.98(br.s.,1H),7.83-7.63(m,6H),7.54(d,J=8.8Hz,1H),7.29(br.s.,1H),6.83(d,J=2.6Hz,1H ), 6.76 (dd, J = 2.2, 8.8Hz, 1H), 5.29 (s, 2H), 4.25-4.04 (m, 4H), 3.65 (s, 2H), 3.48 (t, J = 6.8Hz, 2H), 2.40 (t, J = 7.1Hz, 2H). ESI[M+H]=433.2

[0939]

[0940] 4-(7-(2,6-Diazaspiro[3.5]nonan-2-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 149. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl 2,6-diazaspiro[3.5]nonane-6-carboxylate for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ=8.05(br.s.,1H),7.86-7.69(m,6H),7.58(d,J=8.8Hz,1H),7.35(br.s.,1H),6.74(s,1H),6.67(d,J=8.8Hz,1 H), 5.33 (s, 2H), 3.90 (d, J = 7.8Hz, 2H), 3.80 (d, J = 7.5Hz, 2H), 3.44 (s, 2H), 3.18 (br.s., 2H), 2.02 (d, J = 5.3Hz, 2H), 1.90 (br.s., 2H). ESI[M+H]=447.2

[0941]

[0942] 4-(7-(2,6-diazaspiro[3.4]octan-2-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile, 152. Synthesized using general procedure A substituting 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile (S12) and tert-butyl 2,6-diazaspiro[3.4]octane-6-carboxylate for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 8.05 (s, 1H), 7.85-7.70 (m, 6H), 7.59 (d, J = 8.8Hz, 1H), 7.34 (s, 1H), 6.76 (s, 1H), 6.70 (d,J=8.5Hz,1H),5.33(s,2H),4.09-3.99(m,4H),3.56(s,2H),3.42(t,J=7.3Hz,2H),2.39(t,J=7.3Hz,2H). ESI[M+H]=433.2

[0943]

[0944] 4-(7-((1R,5S,6S)-6-amino-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, 153. The general procedure A was used with 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a The product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile with tert-butyl (1R,5S,6S)-3-azabicyclo[3.1.0]hexan-6-ylcarbamate instead of tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 8.02 (br.s., 1H), 7.86-7.65 (m, 6H), 7.55 (d, J = 8.8Hz, 1H), 7.32 (br.s., 1H), 6.87 (br.s., 1H) ,6.80(d,J=8.8Hz,1H),5.31(s,2H),3.78(d,J=9.3Hz,2H),3.44(d,J=9.3Hz,2H),2.53(br.s.,1H),2.21(br.s.,2H). ESI[M+H]=419.2

[0945]

[0946] 4-(7-(Piperolo[3,4-c]pyrrol-2(1H)-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 154. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl piperillolo[3,4-c]pyrrole-2(1H)-carboxylate for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 8.02 (br.s., 1H), 7.83-7.68 (m, 6H), 7.58 (d, J = 8.8Hz, 1H), 7.31 (br.s., 1H), 6.96 ( br.s.,1H),6.89(d,J=8.8Hz,1H),5.32(s,2H),3.64(d,J=4.9Hz,2H),3.53(br.s.,4H),3.29-3.20(m,4H). ESI[M+H]=433.1

[0947]

[0948] 4-(7-(3-(Methylamino)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 155. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl methyl(pyrrolidin-3-yl)carbamate for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ=8.04(br.s.,1H),7.85-7.65(m,6H),7.60(d,J=8.8Hz,1H),7.33(br.s.,1H),6.92(br.s.,1H),6.85(d,J=8.8Hz,1H),5 .34(s,2H),4.01(br.s.,1H),3.80-3.71(m,1H),3.65(d,J=4.0Hz,2H) ,3.54-3.43(m,1H),2.81(s,3H),2.64-2.49(m,1H),2.37-2.24(m,1H). ESI[M+H]=421.1

[0949]

[0950] (R)-4-(7-(3-(Dimethylamino)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 156. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (R)-N,N-dimethylpyrrolidin-3-amine for tert-butyl piperazine-1-carboxylate. 1 H NMR(400MHz, methanol-d4)δ=8.01(d,J=1.8Hz,1H),7.81-7.69(m,6H),7.61(d,J=8.8Hz,1H),7 .31(d,J=1.5Hz,1H),6.94(d,J=2.3Hz,1H),6.88(dd,J=2.5,8.8Hz,1H),5.34(s,2H),4. 16-4.06(m,1H),3.86(dd,J=7.7,10.7Hz,1H),3.78-3.70(m,1H),3.66(dd,J=6.4,10.7H z,1H),3.54-3.44(m,1H),3.02(s,6H),2.71-2.59(m,1H),2.35(dd,J=8.2,13.2Hz,1H). ESI[M+H]=435.2

[0951]

[0952] 4-(7-(2,6-Diazaspiro[3.3]hept-2-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 157. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 8.03 (br.s., 1H), 7.84-7.65 (m, 6H), 7.55 (d, J = 8.8Hz, 1H), 7.32 (b r.s.,1H),6.73(br.s.,1H),6.65(d,J=8.8Hz,1H),5.30(s,2H),4.33(s,4H),4.18(s,4H). ESI[M+H]=419.2

[0953]

[0954] (S)-4-(7-(3-(Dimethylamino)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 158. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (S)-N,N-dimethylpyrrolidin-3-amine for tert-butyl piperazine-1-carboxylate. 1H NMR(400MHz, methanol-d4)δ=8.01(d,J=1.8Hz,1H),7.81-7.69(m,6H),7.61(d,J=8.8Hz,1H),7 .31(d,J=1.5Hz,1H),6.94(d,J=2.3Hz,1H),6.88(dd,J=2.5,8.8Hz,1H),5.34(s,2H),4. 16-4.06(m,1H),3.86(dd,J=7.7,10.7Hz,1H),3.78-3.70(m,1H),3.66(dd,J=6.4,10.7H z,1H),3.54-3.44(m,1H),3.02(s,6H),2.71-2.59(m,1H),2.35(dd,J=8.2,13.2Hz,1H). ESI[M+H]=435.2

[0955]

[0956] (S)-4-(7-(3-Aminopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 159. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl (S)-pyrrolidin-3-ylcarbamate for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 8.03 (d, J = 2.0Hz, 1H), 7.84-7.69 (m, 6H), 7.61 (d, J = 9.0Hz, 1H), 7.33 (d, J = 1.5Hz, 1H), 6.92 (d, J = 2.5Hz, 1H), 6.85 (d d,J=2.6,8.9Hz,1H),5.35(s,2H),4.16-4.07(m,1H),3.81-3.63(m,2H),3.59-3.49(m,2H),2.61-2.48(m,1H),2.26(dd,J=4.4,8.7Hz,1H). ESI[M+H]=407.1

[0957]

[0958] (R)-4-(7-(3-Aminopyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 161. Synthesized using general procedure A, substituting 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl (R)-pyrrolidin-3-ylcarbamate for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 8.03 (d, J = 2.0Hz, 1H), 7.84-7.69 (m, 6H), 7.61 (d, J = 9.0Hz, 1H), 7.33 (d, J = 1.5Hz, 1H), 6.92 (d, J = 2.5Hz, 1H), 6.85 (d d,J=2.6,8.9Hz,1H),5.35(s,2H),4.16-4.07(m,1H),3.81-3.63(m,2H),3.59-3.49(m,2H),2.61-2.48(m,1H),2.26(dd,J=4.4,8.7Hz,1H). ESI[M+H]=407.1

[0959]

[0960]

[0147] 4-(11-Chloro-7-(piperazin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 164. Synthesized using General Procedure A substituting 4-(7-bromo-11-chloro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile. 1 H NMR (400MHz, methanol-d4) δ = 9.09 (s, 1H), 7.84-7.71 (m, 4H), 7.63 (d, J = 8.8Hz, 1H), 7.34 (d, J = 2.6Hz, 1 H),7.21(dd,J=2.9,9.0Hz,1H),7.15(s,1H),5.31(s,2H),3.61-3.52(m,4H),3.46-3.37(m,4H). ESI[M+H]=442.1

[0961]

[0962] (R)-4-(11-chloro-7-(3-(methylamino)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazo[12-yl)benzonitrile, 166. The general procedure A was used with 4-(7-bromo-11-chloro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazo[12-yl)benzonitrile. The 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile was used instead of 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile and (R)-methyl(pyrrolidin-3-yl)carbamic acid tert-butyl ester was used instead of piperazine-1-carboxylic acid tert-butyl ester. 1 H NMR (400MHz, methanol-d4) δ = 9.06 (br.s., 1H), 7.87-7.79 (m, 2H), 7.78-7.72 (m, 2H) ),7.56(d,J=8.6Hz,1H),7.14(s,1H),6.92(d,J=2.4Hz,1H),6.82(dd,J=2.4, 8.8Hz,1H),5.28(br.s.,2H),4.00(br.s.,1H),3.79-3.58(m,3H),3.52-3.4 1(m,1H),2.86-2.74(m,3H),2.55(dt,J=6.7,14.3Hz,1H),2.34-2.19(m,1H). ESI[M+H]=456.1

[0963]

[0964] (S)-4-(11-chloro-7-(3-(methylamino)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazo[12-yl)benzonitrile, 167. The general procedure A was used with 4-(7-bromo-11-chloro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazo[12-yl)benzonitrile. The 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile was used instead of 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile and (S)-methyl(pyrrolidin-3-yl)carbamate was used instead of tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 9.04 (br.s., 1H), 7.84-7.76 (m, 2H), 7.75-7.68 (m, 2H), 7.54(d,J=8.8Hz,1H),7.12(s,1H),6.90(d,J=2.6Hz,1H),6.80(dd,J=2.6,8.8H z,1H),5.26(br.s.,2H),4.04-3.93(m,1H),3.76-3.55(m,3H),3.45(dt,J=5.7, 9.0Hz, 1H), 2.85-2.73 (m, 3H), 2.61-2.46 (m, 1H), 2.27 (dt, J = 5.4, 13.4Hz, 1H). ESI[M+H]=456.1

[0965]

[0966] 4-(11-chloro-7-(2-methyl-2,6-diazaspiro[3.4]octan-6-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 168. General Procedure C was used with 4-(7-bromo-11-chloro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile. The product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile with tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate instead of tert-butyl (azetidin-3-ylmethyl)(methyl)carbamate. 1 H NMR (400MHz, methanol-d4) δ = 9.03 (s, 1H), 7.83-7.75 (m, 2H), 7.75-7.66 (m, 2H), 7 .51(d,J=8.8Hz,1H),7.11(s,1H),6.80(d,J=3.5Hz,1H),6.72(t,J=6.6Hz, 1H),5.24(br.s.,2H),4.40-4.25(m,2H),4.22-4.07(m,2H),3.70-3.58(m, 2H), 3.45 (td, J=6.7, 13.1Hz, 2H), 2.98 (d, J=5.1Hz, 3H), 2.44-2.33 (m, 2H). ESI[M+H]=482.1

[0967]

[0968] (S)-4-(7-(3-(aminomethyl)pyrrolidin-1-yl)-11-chloro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazo[12-yl)benzonitrile, 169. The general procedure A was used with 4-(7-bromo-11-chloro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazo[12-yl)benzonitrile. The 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile was used instead of 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile and tert-butyl (R)-(pyrrolidin-3-ylmethyl)carbamate was used instead of tert-butyl piperazine-1-carboxylate. 1 H NMR (400 MHz, methanol-d4) δ = 9.04 (br. s., 1H), 7.86-7.79 (m, 2H), 7.78-7.71 (m, 2H), 7.51 (d, J = 8.8 Hz, 1H), 7.13 (s, 1H), 6.80 (d, J = 2.2 Hz, 1H), 6.73 (dd, J = 2.5, 8.7 Hz, 1H), 5.32-5.18(m,2H),3.66-3.50(m,2H),3.47-3.38(m,1H),3.22-3.01(m,3H),2.68( td,J=7.4,14.9Hz,1H), 2.32(dt,J=6.9,11.5Hz,1H), 1.88(qd,J=8.3,12.4Hz,1H). ESI[M+H]=456.1

[0969]

[0970] (S)-4-(11-chloro-7-(3-((dimethylamino)methyl)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazolin-12-yl)benzonitrile, 170. The general procedure C was used with 4-(7-bromo-11-chloro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazolin-12-yl)benzonitrile. The reaction mixture was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile with (R)-(pyrrolidin-3-ylmethyl)carbamate instead of tert-butyl (azetidin-3-ylmethyl)(methyl)carbamate. 1H NMR (400 MHz, methanol-d4) δ = 9.03 (s, 1H), 7.82-7.69 (m, 4H), 7.49 (d, J = 8.8 Hz, 1H), 7.09 (s, 1H), 6.78 (d, J = 2.4 Hz, 1H), 6.71 (dd, J = 2.4, 8.8 Hz, 1H), 5.30-5.17 (m, 2H), 3.72-3.6 1(m,1H),3.52(dt,J=3.1,8.8Hz,1H),3.46-3.36(m,1H),3.34(s,2H),3.20-3.11(m, 1H), 2.97 (s, 6H), 2.91-2.80 (m, 1H), 2.40-2.29 (m, 1H), 1.88 (qd, J = 8.8, 12.3Hz, 1H). ESI[M+H]=484.1

[0971]

[0972] (R)-4-(7-(3-(aminomethyl)pyrrolidin-1-yl)-11-chloro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazo[12-yl)benzonitrile, 171. The general procedure A was used with 4-(7-bromo-11-chloro-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazo[12-yl)benzonitrile. The 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile was used instead of 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile and tert-butyl (S)-(pyrrolidin-3-ylmethyl)carbamate was used instead of tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 9.04 (br.s., 1H), 7.84-7.78 (m, 2H), 7.77-7.70 (m, 2H), 7 .51(d,J=8.8Hz,1H),7.13(s,1H),6.79(d,J=2.2Hz,1H),6.72(dd,J=2.4,8.8Hz ,1H),5.24(br.s.,2H),3.65-3.49(m,2H),3.47-3.39(m,1H),3.22-3.02(m,3H) ,2.74-2.61(m,1H),2.32(dt,J=6.9,11.5Hz,1H),1.88(qd,J=8.3,12.5Hz,1H). ESI[M+H]=456.1

[0973]

[0974] 4-(7-(cis-piperillo[3,2-b]pyrrol-1(2H)-yl)-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 184. General Procedure A was used with 4-(7-bromo-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2 The obtained products were synthesized by using cis-hexahydro-pyrrolo[3,2-b]pyrrole-1(2H)-carboxylic acid tert-butyl ester instead of 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile and cis-hexahydro-pyrrolo[3,2-b]pyrrole-1(2H)-carboxylic acid tert-butyl ester instead of piperazine-1-carboxylic acid tert-butyl ester. 1 H NMR (400MHz, methanol-d4) δ = 7.78 (d, J = 8.8Hz, 1H), 7.76-7.67 (m, 4H), 7.56 (s, 1H), 7. 52(d,J=8.8Hz,1H),7.26(br.s.,1H),6.99(br.s.,1H),6.86(d,J=9.3Hz,1H),5. 33-5.21(m,2H),4.66-4.48(m,2H),3.72-3.56(m,2H),3.44(d,J=4.4Hz,1H),3. 25(br.s.,1H),2.51(d,J=3.1Hz,3H),2.47-2.34(m,2H),2.17(d,J=16.8Hz,2H). ESI[M+H]=447.2

[0975]

[0976] (R)-4-(3-Methyl-7-(3-(methylamino)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 186. Synthesized using general procedure A, substituting 4-(7-bromo-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and tert-butyl (R)-methyl(pyrrolidin-3-yl)carbamate for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 7.81 (s, 1H), 7.74 (q, J = 8.2Hz, 4H), 7.59 (br.s., 1H), 7.53 (d, J = 8.8Hz, 1H), 7.29 (br.s., 1H), 6.97 (br.s., 1H), 6.84 (d ,J=8.8Hz,1H),5.28(br.s.,2H),4.03(br.s.,1H),3.82-3.60(m,3H),3.51(br.s.,1H),2.83(s,3H),2.63-2.48(m,4H),2.32(d,J=5.8Hz,1H). ESI[M+H]=435.2

[0977]

[0978] (S)-4-(3-Methyl-7-(3-(methylamino)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile, 187. Synthesized using general procedure A, substituting 4-(7-bromo-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile and tert-butyl (S)-methyl(pyrrolidin-3-yl)carbamate for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 7.81 (s, 1H), 7.74 (q, J = 8.2Hz, 4H), 7.59 (br.s., 1H), 7.53 (d, J = 8.8Hz, 1H), 7.29 (br.s., 1H), 6.97 (br.s., 1H), 6.84 (d ,J=8.8Hz,1H),5.28(br.s.,2H),4.03(br.s.,1H),3.82-3.60(m,3H),3.51(br.s.,1H),2.83(s,3H),2.63-2.48(m,4H),2.32(d,J=5.8Hz,1H). ESI[M+H]=435.2

[0979]

[0980]

[0147] 4-(3-Methyl-7-(3-((methylamino)methyl)azetidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile, 190. Synthesized using general procedure A substituting 4-(7-bromo-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile and tert-butyl (azetidin-3-ylmethyl)(methyl)carbamate for tert-butyl piperazine-1-carboxylate. 1 H NMR (400MHz, methanol-d4) δ = 7.80 (d, J = 1.3Hz, 1H), 7.78-7.68 (m, 4H), 7.59 (s, 1H), 7.49(d,J=8.8Hz,1H),7.28(d,J=1.3Hz,1H),6.76(d,J=2.3Hz,1H),6.63(dd,J =2.3,8.8Hz,1H),5.25(d,J=4.8Hz,2H),4.19(dt,J=4.0,7.8Hz,2H),3.87-3. 76(m,2H),3.39(d,J=7.3Hz,2H),3.22-3.08(m,1H),2.77(s,3H),2.52(s,3H). ESI[M+H]=435.2

[0981]

[0982] (S)-4-(7-(3-(Aminomethyl)pyrrolidin-1-yl)-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile, 196. Synthesized using general procedure A, substituting 4-(7-bromo-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile and tert-butyl (R)-(pyrrolidin-3-ylmethyl)carbamate for tert-butyl piperazine-1-carboxylate. 1H NMR (400 MHz, methanol-d4) δ = 7.78 (s, 1H), 7.76-7.66 (m, 4H), 7.56 (s, 1H), 7.46 (d, J = 8.8 Hz, 1H), 7.26 (s, 1H), 6.84 (br. s., 1H), 6.73 (d, J = 8.8 Hz, 1H), 5.28-5.18 (m, 2H), 3.67-3.51(m,2H),3.44(d,J=7.9Hz,1H),3.24-3.14(m,1H),3.14-3.03(m,2H),2. 69(td,J=7.1,14.4Hz,1H),2.50(s,3H),2.33(d,J=6.2Hz,1H),1.95-1.82(m,1H). ESI[M+H]=435.2

[0983]

[0984] (S)-4-(7-(3-((dimethylamino)methyl)pyrrolidin-1-yl)-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 197. The general procedure C was used with 4-(7-bromo-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a The product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile with (R)-(pyrrolidin-3-ylmethyl)carbamic acid tert-butyl ester instead of (azetidin-3-ylmethyl)(methyl)carbamic acid tert-butyl ester. 1 H NMR (400MHz, methanol-d4) δ = 7.76 (s, 1H), 7.70 (q, J = 8.4Hz, 4H), 7.55 (s, 1H), 7.45 (d, J = 8.8Hz, 1H) ,7.24(s,1H),6.83(d,J=2.6Hz,1H),6.72(dd,J=2.4,9.0Hz,1H),5.27-5.17(m,2H),3.73-3. 61(m,1H),3.59-3.50(m,1H),3.48-3.38(m,1H),3.37-3.30(m,2H),3.16(dt,J=4.2,8.5Hz,1 H),2.96(s,6H),2.90-2.80(m,1H),2.55-2.44(m,3H),2.41-2.29(m,1H),1.94-1.81(m,1H). ESI[M+H]=463.2

[0985]

[0986] (R)-4-(7-(3-((dimethylamino)methyl)pyrrolidin-1-yl)-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 198. The general procedure C was used with 4-(7-bromo-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a The product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile with (S)-(pyrrolidin-3-ylmethyl)carbamic acid tert-butyl ester instead of (azetidin-3-ylmethyl)(methyl)carbamate. 1 H NMR (400MHz, methanol-d4) δ=7.77(s,1H),7.72(q,J=7.9Hz,4H),7.56(s,1H),7.47(d,J=8. 8Hz,1H),7.26(s,1H),6.85(br.s.,1H),6.73(d,J=8.8Hz,1H),5.23(s,2H),3.73-3. 62(m,1H),3.56(br.s.,1H),3.45(d,J=8.4Hz,1H),3.34(br.s.,2H),3.17(br.s.,1H ),2.97(s,6H),2.92-2.82(m,1H),2.50(s,3H),2.35(br.s.,1H),1.96-1.83(m,1H). ESI[M+H]=463.2

[0987]

[0988]

[0147] 4-(3-Methyl-7-(2,6-diazaspiro[3.4]octan-2-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile, 203. Synthesized using general procedure A substituting 4-(7-bromo-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazopen-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazopen-12-yl)benzonitrile and tert-butyl 2,6-diazaspiro[3.4]octane-6-carboxylate for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 7.80 (s, 1H), 7.74 (q, J = 8.4Hz, 4H), 7.59 (s, 1H), 7.50 (d, J = 8.8Hz, 1H), 7.29 (s, 1H), 6.79 (d, J = 2.5Hz, 1H), 6.66 ( dd,J=2.4,8.7Hz,1H),5.25(d,J=2.8Hz,2H),4.10-3.97(m,4H),3.56(s,2H),3.42(t,J=7.3Hz,2H),2.52(s,3H),2.38(t,J=7.3Hz,2H). ESI[M+H]=447.2

[0989]

[0990] 4-(7-(Piperolo[3,4-c]pyrrol-2(1H)-yl)-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 204. General Procedure A was used with 4-(7-bromo-3-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo-[1 The obtained product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile with tert-butylpiperazine-1-carboxylate and tert-butylpiperazine-1-carboxylate with tert-butylpiperazine-2(1H)-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 7.76 (s, 1H), 7.72 (q, J = 8.1Hz, 4H), 7.54 (s, 1H), 7.49 (d, J = 8.8Hz, 1H), 7.25 (s, 1H), 6.98 (br. s.,1H),6.86(d,J=8.8Hz,1H),5.24(s,2H),3.64(d,J=4.9Hz,2H),3.52(br.s.,4H),3.29-3.21(m,4H),2.50(s,3H). ESI[M+H]=447.2

[0991]

[0992] (R)-4-(7-(3-(Dimethylamino)pyrrolidin-1-yl)-2-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 218. Synthesized using general procedure A substituting 4-(7-bromo-2-methyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (R)-N,N-dimethylpyrrolidin-3-amine for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 7.81-7.68 (m, 6H), 7.57 (d, J = 8.8Hz, 1H), 7.29 (d, J = 1. 8Hz,1H),6.92(d,J=2.4Hz,1H),6.85(dd,J=2.5,8.9Hz,1H),5.32(s,2H),4.1 0(quin,J=7.2Hz,1H),3.84(dd,J=7.6,10.7Hz,1H),3.74-3.60(m,2H),3.51- 3.41(m,1H),3.00(s,6H),2.68-2.57(m,1H),2.49(s,3H),2.40-2.27(m,1H). ESI[M+H]=449.2

[0993]

[0994] 4-(2,3-Dimethyl-7-(3-((methylamino)methyl)pyrrolidin-1-yl)-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 220. Synthesized using general procedure A substituting 4-(7-bromo-2,3-dimethyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c]-[1,4]diazepin-12-yl)benzonitrile and tert-butyl methyl(pyrrolidin-3-ylmethyl)carbamate for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ=7.75-7.67(m,5H),7.41(d,J=8.8Hz,1H),7.21(d,J=1.3H z,1H),6.81(d,J=2.6Hz,1H),6.71(dd,J=2.4,9.0Hz,1H),5.20(s,2H),3.66-3.5 0(m,2H),3.48-3.37(m,1H),3.18-3.13(m,3H),2.75(s,3H),2.74-2.67(m,1H),2 .41(d,J=7.1Hz,6H), 2.32(dd,J=5.1,11.2Hz,1H), 1.88(dd,J=8.4,12.3Hz,1H). ESI[M+H]=463.2

[0995]

[0996] 4-(2,3-Dimethyl-7-(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, 222. The general procedure A was used with 4-(7-bromo-2,3-dimethyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile. The product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazon-12-yl)benzonitrile with tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate instead of tert-butyl piperazine-1-carboxylate. 1H NMR (400 MHz, methanol-d4) δ = 7.76-7.66 (m, 5H), 7.41 (d, J = 8.8 Hz, 1H), 7.21 (d, J = 1.8 Hz, 1H), 6.83 (d, J = 2.2 Hz, 1H), 6.71 (dd, J = 2.6, 8.8 Hz, 1H), 5.26-5.15 (m, 2H), 4.20-4.03 (m, 4H), 3.66-3.60 (m, 2H), 3.46 (t, J = 7.1 Hz, 2H), 2.45-2.35 (m, 8H). ESI [M+H] = 461.2

[0997]

[0998] 4-(7-((R)-3-(Dimethylamino)pyrrolidin-1-yl)-2,3-dimethyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile, 223. Synthesized using General Procedure A substituting 4-(7-bromo-2,3-dimethyl-9H-benzo[e]imidazo[2,1-c]pyrrolo[1,2-a][1,4]diazepin-12-yl)benzonitrile for 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile and (R)-N,N-dimethylpyrrolidin-3-amine for tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ = 7.79-7.65 (m, 5H), 7.46 (d, J = 8.8Hz, 1H), 7.23 (s, 1H) ),6.94(br.s.,1H),6.81(d,J=8.4Hz,1H),5.30-5.17(m,2H),4.09(t,J=6.8 Hz,1H),3.87-3.77(m,1H),3.74-3.58(m,2H),3.46(d,J=6.6Hz,1H),2.98(s ,6H),2.60(br.s.,1H),2.42(d,J=6.2Hz,6H),2.34(dd,J=7.9,13.2Hz,1H). ESI[M+H]=463.2

[0999]

[1000] 4-(3-Methyl-7-(3-((methylamino)methyl)azetidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile, 225. The general procedure A was used with 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. The obtained product was synthesized by replacing 4-(7-bromo-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazolo[3,4-c][1,4]diazepin-12-yl)benzonitrile with tert-butyl (azetidin-3-ylmethyl)(methyl)carbamate instead of tert-butyl piperazine-1-carboxylate. 1H NMR (400MHz, methanol-d4) δ=7.74-7.62(m,5H),7.44(d,J=8.8Hz,1H),7.25(d,J=17.6Hz,1H),6.72(br.s.,1H),6.59(d,J=8.4Hz,1H) ,5.16(br.s.,2H),4.15(d,J=3.1Hz,2H),3.78(d,J=4.9Hz,2H),3.37(d,J=7.1Hz,2H),3.12(d,J=5.7Hz,1H),2.77-2.64(m,6H). ESI[M+H]=436.2

[1001]

[1002] 4-(3-Methyl-7-(3-((methylamino)methyl)pyrrolidin-1-yl)-9H-benzo[e]pyrrolo[1,2-a][1,2,4]triazol...

Claims

1. A compound having a structure of Formula I or a pharmaceutically acceptable salt thereof, in X 1 It is N; X 2 It's CR 5 or N; X 3 It's CR 5 or N; X 5 is C(CH3) or CH; X 6 and X 7 Each occurrence is independently CH or N; R 1 Each occurrence is independently H, fluoro, or -CN; R 2 It is H or chlorine; R 3 is H or CH3; R 5 It is H; Z is X 20 It's CR 24 R 26 or NH; X 21 It is NH; R 20 、R 21 、R 22 and R 23 Each independently represents H, fluorine, hydroxyl, amino, C1 alkyl, amino C 1-2 Alkyl, C 1-2 Alkylamino C 1-2 Alkyl or C1 alkylamino; or R 23 and R 20 Combine to form a nitrogen-containing 5-membered ring; R 21 and R 20 Combine to form a nitrogen-containing 5-membered ring; or R 21 and R 22 Combine to form a nitrogen-containing 4-membered ring; R 24 and R 26 are each independently H, amino, aminoC1alkyl, C1alkylaminoC1alkyl or C1alkylamino; or R 24 With R 26 Combine to form a nitrogen-containing 4-membered ring; R 25 and R 27 Each is independently H, amino, C1 alkyl, C1 alkylaminoC1 alkyl or C1 alkylamino; R 28 and R 29 are each independently H, amino, aminoC1 alkyl, C1 alkylaminoC1 alkyl or C1 alkylamino; or R 28 and R 29 Combined to form a nitrogen-containing 4-membered or 5-membered ring; and n is an integer from 0 to 5.

2. The compound according to claim 1, wherein the compound has a structure of Formula II, or a pharmaceutically acceptable salt thereof, in R 1a is H, fluorine, or -CN; and R 1b is H, fluorine or -CN.

3. The compound according to claim 1, wherein X 1 is N; and X 2 and X 3 It is CH.

4. The compound according to claim 1, wherein X 1 and X 2 is N; and X 3 It is CH.

5. The compound according to claim 1, wherein X 1 and X 3 is N; and X 2 It is CH.

6. The compound according to claim 1, wherein X 1 、X 2 and X 3 It's N.

7. The compound according to claim 1, wherein Z is 8. The compound according to claim 7, wherein Z is 。 9. The compound according to claim 7, wherein Z is 。 10. The compound according to claim 1, wherein Z is 11. The compound according to claim 10, wherein Z is 12. The compound according to claim 1, wherein Z is 13. The compound according to claim 12, wherein Z is 14. The compound according to claim 1, wherein Z is 15. The compound according to claim 1, wherein R 1 It is -CN.

16. The compound according to claim 2, wherein R 1a and R 1b Each occurrence is independently H or -CN.

17. The compound according to claim 2, wherein R 1a and R 1b Each occurrence is H or fluorine.

18. The compound according to claim 1, wherein R 2 It’s H.

19. The compound according to claim 1, wherein R 2 It's chlorine.

20. The compound according to claim 1, wherein R 3 It is C 1-6 alkyl.

21. The compound according to claim 1, wherein R 3 It’s H.

22. The compound of claim 1, wherein n is 0, 1 or 2.

23. A compound selected from: or a pharmaceutically acceptable salt thereof.

24. A pharmaceutical composition comprising a compound according to any one of claims 1 to 23 and a pharmaceutically acceptable carrier.

25. Use of a compound according to any one of claims 1 to 23 in the preparation of a medicament for treating or preventing a mitogen-activated protein kinase-activated protein kinase-2 (MK2)-related disorder.

26. The use according to claim 25, wherein the MK2-related disorder is an inflammatory disorder or cancer.

27. The use according to claim 26, wherein the cancer is a KRAS-dependent or BRAF-dependent cancer.

28. Use of a compound according to any one of claims 1 to 23 in combination with one or more other chemotherapeutic agents for the preparation of a medicament for the treatment or prevention of a mitogen-activated protein kinase-activated protein kinase-2 (MK2)-related disorder.

29. The use according to claim 28, wherein the other chemotherapeutic agent is a CHK1 inhibitor or cisplatin.

30. The use according to claim 29, wherein the CHK1 inhibitor is PF477736 or LY2603618.

31. Use of the compound according to any one of claims 1 to 23 in the preparation of a medicament for inhibiting the proliferation of cancer cells.

32. Use of a compound according to any one of claims 1 to 23 in the preparation of a medicament for inhibiting MK2 activity in a cell.

33. Use of a compound according to any one of claims 1 to 23 in the preparation of a medicament for treating or preventing a metabolic disorder.

34. Use according to claim 33, wherein the metabolic disorder is diabetes, insulin resistance, obesity or metabolic syndrome.

35. Use according to claim 34, wherein the diabetes is type I, type II or gestational diabetes.

36. The use according to claim 33, wherein the metabolic disorder is treated or prevented by affecting glycogenolysis or gluconeogenesis.

37. The use of claim 33, wherein the metabolic disorder is treated or prevented by reducing hepatic glucose production, hyperglycemia, fatty liver, insulin resistance, insulin resistance-associated inflammation, insulin resistance-associated dyslipidemia, or any combination thereof.

38. Use of a compound according to any one of claims 1 to 23 in combination with one or more other antidiabetic agents for the preparation of a medicament for the treatment or prevention of diabetes or insulin resistance.

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