Piperidinol-benzodiazepine compounds with antiproliferative activity

Abstract

The present invention relates to pyridobenzo diazepines (PDDs) comprising three fused 6-7-6-membered rings attached to an aromatic group and pharmaceutically acceptable salts thereof, which are useful as pharmaceuticals, such as anti-proliferatives. The PDDs can be represented by Formula (I): and salts and solvates thereof.

Description

[0001] This application is filed on August 19, 2016, and is titled "PIPERIDIZED BENZODIAZEPHILATA WITH ANTI-PROLIFERATIVE ACTIVITY." This is a divisional application of Chinese patent application No. 201680060201.8 entitled "Compound". Field of the Invention

[0002] The present invention relates to pyridobenzodiazepines comprising three fused 6-7-6-membered rings (PDD). In particular, the present invention relates to compounds comprising a PDD group linked to an aromatic group via the A ring, and to pharmaceutically acceptable salts thereof, which are useful as medicaments, in particular as antiproliferative agents. Background of the Invention

[0003] Pyridobenzodiazepine (PDD) is pyrrolobenzodiazepine (PBD) related structures. PBDs are a group of compounds, some of which have been shown to be sequence-selective DNA minor-groove binders. PBDs were originally discovered in Streptomyces species (1-5). They are tricyclic in nature and consist of an anthranilic acid (A ring), a diazepine The pyrrolidine rings consist of fused 6-7-5-membered rings of pyrrolidine (Ring B) and pyrrolidine (Ring C) (3). They are characterized by an electrophilic N10=C11 imine group (shown below) or a hydrated equivalent, carbinolamine [NH-CH(OH)] or carbinolamine alkyl ether [NH-CH(OR, where R=alkyl)], which can form a covalent bond with the C2-amino group of guanine in DNA to form DNA adducts (6).

[0004]

[0005] Due to the right-handed longitudinal rotation caused by the chiral C11a-position with the (S)-configuration, the natural product interacts with excellent fit (i.e., good "isohelicity") in the minor groove of the DNA helix (6). DNA adducts have been reported to inhibit many biological processes, including the binding of transcription factors (7-9) and the function of enzymes such as endonucleases (10, 11) and RNA polymerases (12). PBD monomers (e.g., anthramycin) have been shown by footprinting (6), NMR (13, 14), molecular modeling (15), and X-ray crystallography (16) to span three base pairs and to have a thermodynamic preference for the sequence 5'-Pu-G-Pu-3' (where Pu = purine and G is the reacting guanine (17)) and a kinetic preference for Py-5-Py (where Py = pyrimidine).

[0006] PBDs are thought to interact with DNA primarily by localizing to low-energy binding sequences (i.e., 5'-Pu-G-Pu-3' triplet) via van der Waals, hydrogen bonding, and electrostatic interactions (7). Thus, once in place, nucleophilic attack by the exocyclic C2-amino group of the central guanine occurs to form a covalent adduct (7). Once bound, the PBD remains anchored in the minor groove of the DNA, avoiding DNA repair by causing negligible deformation of the DNA helix (16). The ability of PBDs to form adducts in the minor groove and cross-link DNA makes them interfere with DNA processing and therefore have the potential to be used as antiproliferative agents.

[0007] Many monomeric PBD structures have been isolated from Streptomyces species, including the first PBD anthramycin (18), tomamycin (19), and more recently usabamycin (20) from marine sediment Streptomyces species. This has led to the development of a wide range of synthetic analogs that have been reviewed (1, 21). More recently, a number of monomeric PBD structures linked to pyrrole and imidazole via their C8 position have been reported, WO 2007 / 039752, WO 2013 / 164593 (22-27).

[0008] WO 2010 / 091150 discloses dimers of 6-7-6 ring systems connected via their A-rings. WO 2015 / 028850 discloses dimers of 6-7-5 ring systems PBDs connected via a phosphine oxide-containing linker connected to their aromatic A-rings. In addition, WO 2015 / 028850 discloses dimer compounds containing a 6-7-6 ring system connected via a key phosphine oxide-containing linker.

[0009] Various PBDs have been shown to act as cytotoxic agents in vitro, e.g., WO 00 / 12508, WO 2004 / 087711, and to have anti-tumor effects in vivo in animal tumor models, e.g., WO 2011 / 117882, WO 2013 / 164593. In addition, the C8 / C8'-linked PBD dimer SJG-136 (28, 29) has completed Phase I clinical trials for leukemia and ovarian cancer (30) and has shown sufficient therapeutic benefit to proceed to Phase II studies.

[0010]

[0011]

[0012] However, phase I clinical evaluation of SJG-136 revealed that the drug produced several adverse reactions including lower limb edema and fatigue (31).

[0013] Therefore, there is a need for additional compounds related to PBD that have therapeutic activity for the treatment of various proliferative diseases.

[0014] This application reports pyridobenzodiazepine (PDD), which is related to PBD but contains an enlarged 6-membered C ring compared to the 5-membered C ring of PBD. The present inventors have found that PDD conjugates provide properties such as cytotoxicity and DNA binding, which result in potent compounds.

[0015] The present invention seeks to overcome the problems associated with the prior art. SUMMARY OF THE INVENTION

[0016] The present invention provides compounds of formula (I):

[0017]

[0018] and its salts and solvates,

[0019] in;

[0020] Dashed lines indicate the optional presence of double bonds between one or more of C1 and C2, C2 and C3, and C3 and C4;

[0021] R1 is selected from R7, =CH2, =CH-(CH2) m -CH3, =O, (CH2) m -OR7, (CH2) m -CO2R7, (CH2) m -NR7R8, O-(CH2) n -NR7R8, NH-C(O)-R7, O-(CH2)n -NH-C(O)-R7, O-(CH2) n -C(O)-NH-R7, (CH2) m -SO2R7, O-SO2R7, (CH2) m -C(O)R7 and (CH2) m -C(O)NR7R8;

[0022] R2 is selected from R9, =CH2, =CH-(CH2) r -CH3, =O, (CH2) r -OR9, (CH2) r -CO2R9, (CH2) r -NR9R 10 、O-(CH2) s -NR9R 10 、NH-C(O)-R9、O-(CH2) s -NH-C(O)-R9, O-(CH2) s -C(O)-NH-R9, (CH2) r -SO2R9, O-SO2R9, (CH2) r -COR9 and (CH2) r -C(O)NR9R 10 ;

[0023] R3 is selected from H, C 1-12 Alkyl and CH2Ph;

[0024] R4 is selected from phenyl and C 5-9 Heteroaryl, the substituent is selected from OH, C 1-6 Alkyl, OC 1-6 Alkyl, (CH2) j -CO2R 11 、O-(CH2) k -NR 11 R 12 、(CH2) j -NR 11 R 12 、C(=O)-NH-(CH2) k -NR 11 R 12 ; C(=O)-NH-R 24 and C(=O)-NH-(CH2) k -C(=NH)UR 11 R 12 ; Provided that the optionally substituted C 5-9 Heteroaryl is not indolyl;

[0025] R 19 Selected from H and (CH2) t -NR 20 R 21 ;

[0026] Y1 is N or CH;

[0027] Y2 is N or CH; and wherein at least one of Y1 and Y2 is CH;

[0028] p is 0 or 1;

[0029] j, m, r and t are independently selected from integers from 0 to 6;

[0030] k, n and s are independently selected from integers from 1 to 6;

[0031] X1 is selected from O, S, NR 13 , CR 13 R 14 , CR 13 R 14 O, C(=O), C(=O)NR 13 NR 13 C(=O), OC(O) and C(O)-O;

[0032] L is selected from amino acids, peptide chains having 2 to 6 amino acids, alkylene chains containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, paraformaldehyde chains -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 -, the chain may be substituted by O, S and / or NH groups and / or C 3-9 one or more interruptions in the heteroaryl and / or phenylene groups;

[0033] X2 is selected from O, S, NR 15 , CR 15 R 16 , CR 15 R 16 O, C(=O), C(=O)NR 15 NR 15 C(=O), OC(O) and C(O)-O or absent;

[0034] q is selected from 0, 1, 2, 3, 4, 5 and 6;

[0035] A is selected from:

[0036]

[0037] For each A1 group, one of Y3 and Y4 is independently selected from NR 17 , S and O; and the other of Y3 and Y4 is CH; and Y5 is independently selected from CH, N, S and COH; and

[0038] For each A2 group, one of Y6 and Y7 is independently selected from N and CH; and the other of Y6 and Y7 is CH;

[0039] R7 and R9 are independently selected from H, C 1-12 Alkyl, C 5-9 Heteroaryl, C 6-15 Heteroarylalkyl, phenyl and C 7-12 Aralkyl groups; wherein the heteroaryl, heteroarylalkyl, phenyl and aralkyl groups are optionally substituted with up to three optional substituents selected from C 1-6 Alkyl, OH, OC 1-6 alkyl;

[0040] R 24 is phenyl optionally substituted with up to three optional substituents selected from OH, C 1-6 Alkyl, OC 1-6 Alkyl, (CH2) j -CO2R 11 、O-(CH2) k -NR 11 R 12 、(CH2) j -NR 11 R 12 、C(=O)-NH-(CH2) k -NR 11 R 12 and C(=O)-NH-(CH2) k -C(=NH)UR 11 R 12 ;

[0041] R8, R 10 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 20 and R 21 Independently selected from H and C 1-6 alkyl;

[0042] or:

[0043] (i) R5 and R6 together form a double bond;

[0044] (ii) R5 is H and R6 is OH; or

[0045] (iii) R5 is H and R6 is OC 1-6 alkyl;

[0046] The condition is that when p is 0 and A is A1, then:

[0047] (a) for at least one A1 group, one of Y3 and Y4 is selected from S and O; or

[0048] (b) for at least one A1 group, Y5 is S; or

[0049] (c) R4 is not pyrrolyl, imidazolyl, optionally substituted pyrrolyl or optionally substituted imidazolyl.

[0050] The present invention provides compounds of formula (I):

[0051]

[0052] and its salts and solvates,

[0053] in;

[0054] Dashed lines indicate the optional presence of double bonds between one or more of C1 and C2, C2 and C3, and C3 and C4;

[0055] R1 is selected from R7, =CH2, =CH-(CH2) m -CH3, =O, (CH2) m -OR7, (CH2) m -CO2R7, (CH2) m -NR7R8, O-(CH2) n -NR7R8, NH-C(O)-R7, O-(CH2) n -NH-C(O)-R7, O-(CH2) n -C(O)-NH-R7, (CH2) m -SO2R7, O-SO2R7, (CH2) m -C(O)R7 and (CH2) m -C(O)NR7R8;

[0056] R2 is selected from R9, =CH2, =CH-(CH2) r -CH3, =O, (CH2) r -OR9, (CH2) r -CO2R9, (CH2) r -NR9R 10 、O-(CH2) s-NR9R 10 , NH-C(O)-R9, O-(CH2) s -NH-C(O)-R9, O-(CH2) s -C(O)-NH-R9, (CH2) r -SO2R9, O-SO2R9, (CH2) r -COR9and (CH2) r -C(O)NR9R 10 ;

[0057] R3is selected from H, C 1-12 alkyl and CH2Ph;

[0058] R4is selected from phenyl and C 5-9 heteroaryl optionally substituted with up to three optional substituents selected from the group consisting of OH, C 1-6 alkyl, OC 1-6 alkyl, (CH2) j -CO2R 11 , O-(CH2) k -NR 11 R 12 , (CH2) j -NR 11 R 12 , C(=O)-NH-(CH2) k -NR 11 R 12 ; C(=O)-NH-C6H4-(CH2) j -R 18 and C(=O)-NH-(CH2) k -C(=NH)NR 11 R 12 ; provided that the optionally substituted C 5-9 heteroaryl is not indolyl;

[0059] R 19 is selected from H and (CH2) t -NR 20 R 21 ;

[0060] Y1is N or CH;

[0061] Y2is N or CH; and wherein at least one of Y1and Y2is CH;

[0062] p is 0 or 1 ;

[0063] j, m, r and t are independently selected from integers ranging from 0 to 6;

[0064] k, n and s are independently selected from integers ranging from 1 to 6;

[0065] X1 is selected from O, S, NR 13 , CR 13 R 14 , CR 13 R 14 O, C(=O), C(=O)NR 13 NR 13 C(=O), OC(O) and C(O)-O;

[0066] L is selected from amino acids, peptide chains having 2 to 6 amino acids, alkylene chains containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, paraformaldehyde chains -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 -, the chain may be substituted by O, S and / or NH groups and / or C 3-9 interrupted by one or more of heteroarylene and / or phenylene;

[0067] X2 is selected from O, S, NR 15 , CR 15 R 16 , CR 15 R 16 O, C(=O), C(=O)NR 15 NR 15 C(=O), OC(O) and C(O)-O or absent;

[0068] q is selected from 0, 1, 2, 3, 4, 5 and 6;

[0069] A is selected from:

[0070]

[0071] For each A1 group, one of Y3 and Y4 is independently selected from NR 17 , S and O; and the other of Y3 and Y4 is CH; and Y5 is independently selected from CH, N, S and COH; and

[0072] For each A2 group, one of Y6 and Y7 is independently selected from N and CH; and the other of Y6 and Y7 is CH;

[0073] R7 and R9 are independently selected from H, C 1-12 Alkyl, C 5-9 Heteroaryl, C 6-15 Heteroarylalkyl, phenyl and C 7-12 Aralkyl groups; wherein the heteroaryl, heteroarylalkyl, phenyl and aralkyl groups are optionally substituted with up to three optional substituents selected from C1-6 Alkyl, OH, OC 1-6 alkyl;

[0074] R 18 Selected from CO2R 11 and NR 11 R 12 ;

[0075] R8, R 10 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 20 and R 21 Independently selected from H and C 1-6 alkyl;

[0076] or:

[0077] (i) R5 and R6 together form a double bond;

[0078] (ii) R5 is H and R6 is OH; or

[0079] (iii) R5 is H and R6 is OC 1-6 alkyl;

[0080] The condition is that when p is 0 and A is A1, then:

[0081] (a) for at least one A1 group, one of Y3 and Y4 is selected from S and O; or

[0082] (b) for at least one A1 group, Y5 is S; or

[0083] (c) R4 is not pyrrolyl, imidazolyl, optionally substituted pyrrolyl or optionally substituted imidazolyl.

[0084] In a further aspect, provided are compounds of formula (I) and salts and solvates thereof for use in methods of treatment.

[0085] In a further aspect, there is provided a compound of formula (I) and salts and solvates thereof for use as a medicament.

[0086] In a further aspect, provided are compounds of formula (I) and salts and solvates thereof for use in the treatment of a proliferative disease.

[0087] In a further aspect, a pharmaceutical composition is provided comprising a compound of formula (I) and salts and solvates thereof and a pharmaceutically acceptable excipient, carrier or diluent.

[0088] In a further aspect, the present application provides the use of a compound of Formula (I) and salts and solvates thereof in the manufacture of a medicament for the treatment of a proliferative disease.

[0089] In a further aspect, the present application provides a method of treating a patient having a proliferative disease, comprising administering to the patient a therapeutically effective amount of a compound of Formula (I) and salts and solvates thereof or a pharmaceutical composition of the present application.

[0090] In a further aspect, depending upon the condition to be treated, a compound of Formula (I) and salts and solvates thereof can be administered alone or in combination with other therapies, either simultaneously or sequentially.

[0091] A pharmaceutical composition of the present application can further comprise one or more (e.g., two, three, or four) additional active agents.

[0092] In a further aspect, a compound of Formula (I) and salts and solvates thereof can be directly or indirectly linked to a targeting agent (e.g., an antibody, antibody fragment, hormone, etc.) to provide a targeted conjugate. A targeted conjugate of the present disclosure can contain one or more compounds of Formula (I) (or salts and solvates thereof). A variety of targeted conjugates are known in the art and can be used with a compound of Formula (I) and salts and solvates thereof. For example, in a particular aspect, the targeted conjugate is an antibody-drug conjugate in which one or more compounds of Formula (I) are directly or indirectly linked to an antibody. Thus, a compound of Formula (I) and salts and solvates thereof can be used as a payload on a targeted conjugate.

[0093] definition

[0094] The following abbreviations are used throughout the specification: Ac, acetyl; Alloc, allyloxycarbonyl; BAIB, bis(acetyloxy)iodobenzene / (diacetyloxyiodo)benzene; Boc, tert-butyloxycarbonyl; BPD, benzo pyrrido diazecine (benzopyrridodiazecine); CBz, benzyloxycarbonyl; DBU, 1,8-diazabicyclo[5.4.0]undec-7-ene; DHP, dihydropyran; DMAP, 4-dimethylaminopyridine; DMF, dimethylformamide; DMSO, dimethyl sulfoxide; EDCI, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide; Et, ethyl; Et20, diethyl ether; EtOAc, ethyl acetate; EtOH, ethanol; HATU, (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxyhexafluorophosphate); HMDST, hexamethyldisilathione; iBu, isobutyl; KOtBu, potassium tert-butoxide; L-Selectride lithium tri-sec-butyl(hydride)borate; Me, methyl; MeOH, methanol; PBD, pyrrolo[2,1-c][1,4]benzodiazepine ; PDD, pyridobenzodiazepine ; PIFA, bis(trifluoroacetoxy)phenyliodide(III); Ph, phenyl; p-TSA / PTSA, p-toluenesulfonic acid; Pyr, pyridine; TBAF, tetrabutylammonium fluoride; TBS-Cl / TBDMSCl, tert-butyldimethylchlorosilane; TEA, triethylamine; TEMPO, (2,2,6,6-tetramethyl-piperidin-1-yl)oxy; TFA, trifluoroacetic acid; THF, tetrahydrofuran; THP, tetrahydropyranyl; Troc, 2,2,2-trichloroethyl carbonate, and Ts (toluenesulfonate), p-toluenesulfonic acid.

[0095] When used in conjunction with a chemical substituent or moiety (e.g., alkyl), "substituted" means that one or more hydrogen atoms of the substituent or moiety have been replaced with one or more non-hydrogen atoms or groups, provided that valence requirements are met and the substitution results in a chemically stable compound.

[0096] "Optionally substituted" refers to a parent group that may be unsubstituted or substituted with one or more substituents. Suitably, unless otherwise stated, when there are optional substituents, the optionally substituted parent group comprises one to three optional substituents. When a group can be "optionally substituted with up to three groups", this means that the group can be substituted with 0, 1, 2 or 3 optional substituents. When a group can be "optionally substituted with one or two optional substituents", this means that the group can be substituted with 0, 1 or 2 optional substituents. Suitably, a group can be optionally substituted with 0 or 1 optional substituents.

[0097] "Independently selected from" is used in the context of, for example, "R1 and R2 are independently selected from H, C 1-12 alkyl, phenyl...” and means that each instance of a functional group (e.g., R1) is selected from the listed options independently of any other instances of R1 or R2 in the compound. Thus, for example, C 1-12 Alkyl; phenyl may be selected for the next instance of R1 in the compound; and H may be selected for the first instance of R2 in the compound.

[0098] C 1-12 Alkyl: refers to a straight or branched chain saturated hydrocarbon group, usually having 1 to 12 carbon atoms; more preferably C 1-7 Alkyl; more preferably C1-6 Alkyl. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentan-1-yl, pentan-2-yl, pentan-3-yl, 3-methylbutan-1-yl, 3-methylbutan-2-yl, 2-methylbutan-2-yl, 2,2,2-trimethylethan-1-yl, n-hexyl, n-heptyl, and the like.

[0099] "Hydrocarbylene" refers to a divalent group derived from an alkane which may be straight or branched chain, for example -CH2CH2CH2CH2-.

[0100] "Aryl" refers to fully unsaturated monocyclic, bicyclic, and polycyclic aromatic hydrocarbons (e.g., C 6-14 Aryl refers to an aromatic radical having 6 to 14 carbon atoms as ring members. The aromatic radical may be attached to the parent radical or to the substrate at any ring atom and may include one or more non-hydrogen substituents unless such attachment or substitution would violate valence requirements. Examples of aromatic radicals include phenyl.

[0101] “C 7-12 "Aralkyl" refers to an arylalkyl group having 7 to 12 carbon atoms and comprising an alkyl group substituted with an aryl group. Suitably, the alkyl group is C 1-6 The alkyl group and the aryl group is phenyl. 7-12 Examples of aralkyl groups include benzyl and phenethyl. 7-12 The aralkyl group may be optionally substituted and optionally substituted C 7-12 An example of an aralkyl group is 4-methoxybenzyl.

[0102] “C 5-9 "Heteroaryl" refers to an unsaturated monocyclic or bicyclic aromatic group containing from 5 to 9 ring atoms (whether carbon or heteroatoms), 1 to 5 of which are ring heteroatoms. Suitably, any monocyclic heteroaryl ring has from 5 to 6 ring atoms and from 1 to 3 ring heteroatoms. Suitably, each ring heteroatom is independently selected from nitrogen, oxygen and sulfur. Bicyclic includes fused ring systems and particularly includes bicyclic groups in which a monocyclic ring containing 5 ring atoms is fused to a benzene ring. The heteroaryl group may be attached to the parent group or to the substrate at any ring atom and may include one or more non-hydrogen substituents unless such attachment or substitution would violate valence requirements or produce a chemically unstable compound.

[0103] Examples of monocyclic heteroaryl groups include, but are not limited to, those derived from:

[0104] N1: pyrrole, pyridine;

[0105] O1: Furan;

[0106] S1: thiophene;

[0107] N1O1: Azoles, isocyanates Azoles, isocyanates oxazine;

[0108] N2O1: Oxazolyl (e.g., 1-oxa-2,3-oxazolyl, 1-oxa-2,4-oxazolyl, 1-oxa-2,5-oxazolyl, 1-oxa-3,4-oxazolyl);

[0109] N3O1: triazole;

[0110] N1S1: thiazole, isothiazole;

[0111] N2: imidazole, pyrazole, pyridazine, pyrimidine, pyrazine;

[0112] N3: triazole, triazine; and

[0113] N4: tetrazole.

[0114] Examples of heteroaryl groups containing fused rings include, but are not limited to, those derived from:

[0115] O1: benzofuran, isobenzofuran;

[0116] N1: indole, isoindole, indolizine, isoindoline;

[0117] S1: thionaphthene;

[0118] N1O1: benzo azole, benzyl isocyanate azole;

[0119] N1S1: benzothiazole;

[0120] N2: benzimidazole, indazole;

[0121] O2: benzodioxole;

[0122] N2O1: benzofurazan;

[0123] N2S1: benzothiadiazole;

[0124] N3: benzotriazole; and

[0125] N4: purines (e.g., adenine, guanine), pteridines;

[0126] "Heteroarylene" refers to a divalent radical derived from a heteroaryl group, for example, pyridylene-(C5H3N)-.

[0127] “C 6-15 "Heteroarylalkyl" refers to an alkyl group substituted with a heteroaryl group. Suitably, the alkyl group is C 1-6 The alkyl group and the heteroaryl group are as defined above C 5-9 Heteroaryl. C 6-15 Examples of heteroarylalkyl groups include pyrrol-2-ylmethyl, pyrrol-3-ylmethyl, pyrrol-4-ylmethyl, pyrrol-3-ylethyl, pyrrol-4-ylethyl, imidazol-2-ylmethyl, imidazol-4-ylmethyl, imidazol-4-ylethyl, thien-3-ylmethyl, furan-3-ylmethyl, pyridin-2-ylmethyl, pyridin-2-ylethyl, thiazol-2-ylmethyl, thiazol-4-ylmethyl, thiazol-2-ylethyl, pyrimidin-2-ylpropyl, and the like.

[0128] Nitrogen protecting group

[0129] Nitrogen protecting groups are well known in the art.

[0130] A preferred nitrogen protecting group is a carbamate protecting group having the following general formula:

[0131]

[0132] A large number of possible carbamate nitrogen protecting groups are listed in Wuts, PGM and Greene, TW, Protective Groups in Organic Synthesis, 4th edition, Wiley-Interscience, 2007, pages 706 to 771, and in P. Kocienski, Protective Groups, 3rd edition (2005), which are incorporated herein by reference.

[0133] Particularly preferred protecting groups include Alloc (allyloxycarbonyl), Troc (2,2,2-trichloroethyl carbonate), Teoc [2-(trimethylsilyl)ethoxycarbonyl], BOC (tert-butoxycarbonyl), Doc (2,4-dimethylpent-3-yloxycarbonyl), Hoc (cyclohexyloxy-carbonyl), TcBOC (2,2,2-trichloro-tert-butoxycarbonyl), Fmoc (9-fluorenylmethoxycarbonyl), 1-Adoc (1-adamantyloxycarbonyl) and 2-Adoc (2-adamantyloxycarbonyl).

[0134] Hydroxyl protecting group

[0135] Hydroxyl protecting groups are well known in the art, and many suitable groups are described in Wuts, PGM and Greene, TW, Protective Groups in Organic Synthesis, 4th edition, Wiley-Interscience, 2007, pp. 16-366, and in P. Kocienski, Protective Groups, 3rd edition (2005), which are incorporated herein by reference.

[0136] Classes of particular interest include silyl ethers, methyl ethers, alkyl ethers, benzyl ethers, esters, benzoates, carbonates, and sulfonates.

[0137] Particularly preferred protecting groups include THP (tetrahydropyranyl ether).

[0138] "Compounds of formula (I) and salts and solvates thereof" refers to compounds of formula (I); salts of compounds of formula (I); solvates of compounds of formula (I); and solvates of salts of compounds of formula (I).

[0139] "Drug," "drug substance," "active pharmaceutical ingredient," and the like refer to a compound (eg, a compound of Formula (I) and the compounds specifically identified above) that can be used to treat a subject in need of treatment.

[0140] "Excipient" refers to any substance that may affect the bioavailability of a drug but is otherwise pharmacologically inactive.

[0141] "Pharmaceutically acceptable" substances are those substances which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of subjects without undue toxicity, irritation, allergic response, etc., commensurate with a reasonable benefit-to-risk ratio, and effective for their intended use.

[0142] A "pharmaceutical composition" refers to a combination of one or more drug substances and one or more excipients.

[0143] As used herein, the term "subject" refers to a human or non-human mammal. Examples of non-human mammals include livestock animals such as sheep, horses, cattle, pigs, goats, rabbits, and deer; and companion animals such as cats, dogs, rodents, and horses.

[0144] The "therapeutically effective amount" of a drug refers to the amount of the drug or composition that effectively treats a subject and thereby produces the desired therapeutic, ameliorative, inhibitory or preventive effect. The therapeutically effective amount may depend on the subject's weight and age, the route of administration, and the like.

[0145] "Treat," "treat ...

[0146] "Treatment" means the act of treating as defined above.

[0147] As used herein, the term "comprising" means "including at least in part" and is intended to be inclusive or open-ended. When interpreting each statement in this specification that includes the term "comprising," there may also be features, elements, and / or steps other than the one or those features, elements, and / or steps that begin with the term. Related terms, such as "comprise" and "comprises," are to be interpreted in the same manner.

[0148] R1

[0149] R1 is selected from R7, =CH2, =CH-(CH2) m -CH3, =O, (CH2) m -OR7, (CH2) m -CO2R7, (CH2) m -NR7R8, O-(CH2) n -NR7R8, NH-C(O)-R7, O-(CH2) n -NH-C(O)-R7, O-(CH2) n -C(O)-NH-R7, (CH2) m -SO2R7, O-SO2R7, (CH2) m -C(O)R7 and (CH2) m -C(O)NR7R8. For wherein R1 is selected from =CH2, =CH-(CH2) m -CH3 and =O, the carbon of the C-ring to which it is attached cannot have an optional double bond in order to meet the compound requirements of the molecule. For example, if R1 is =CH2 and is located at the C1 position of the C-ring adjacent to the fused carbon of the C-ring, and R2 is H, then the resulting compound of formula (I) can be represented as:

[0150]

[0151] Suitably, R1 is selected from R7, (CH2) m -OR7, (CH2) m -CO2R7, (CH2) m-NR7R8, O-(CH2) n -NR7R8, NH-C(O)-R7, O-(CH2) n -NH-C(O)-R7, O-(CH2) n -C(O)-NH-R7, (CH2) m -SO2R7, O-SO2R7, (CH2) m -C(O)R7 and (CH2) m -C(O)NR7R8.

[0152] Suitably, R1 is selected from R7, (CH2) m -OR7, (CH2) m -CO2R7, (CH2) m -NR7R8, O-(CH2) n -NR7R8, NH-C(O)-R7, O-(CH2) n -NH-C(O)-R7, O-(CH2) n -C(O)-NH-R7, (CH2) m -C(O)R7 and (CH2) m -C(O)NR7R8.

[0153] Suitably, R1 is selected from R7, OR7, CO2R7, NR7R8, NH-C(O)-R7, O-(CH2) n -NH-C(O)-R7, O-(CH2) n -C(O)-NH-R7, C(O)R7 and C(O)NR7R8.

[0154] Suitably, R1 is selected from R7, OR7, CO2R7, O-(CH2) n -NH-C(O)-R7, O-(CH2) n -C(O)-NH-R7, C(O)R7 and C(O)NR7R8.

[0155] Suitably, R1 is selected from R7, O-(CH2) n -NH-C(O)-R7 and O-(CH2) n -C(O)-NH-R7.

[0156] In some embodiments, R1 is H.

[0157] R2

[0158] R2 is selected from R9, (CH2) r -OR9, (CH2) r-CO2R9, (CH2) r -NR9R 10 、O-(CH2) s -NR9R 10 、NH-C(O)-R9、O-(CH2) s -NH-C(O)-R9, O-(CH2) s -C(O)-NH-R9, (CH2) r -SO2R9, O-SO2R9, (CH2) r -COR9 and (CH2) r -C(O)NR9R 10 .

[0159] Suitably, R2 is selected from R9, (CH2) r -OR9, (CH2) r -CO2R9, (CH2) r -NR9R 10 、O-(CH2) s -NR9R 10 、NH-C(O)-R9、O-(CH2) s -NH-C(O)-R9, O-(CH2) s -C(O)-NH-R9, (CH2) r -COR9 and (CH2) r -C(O)NR9R 10 .

[0160] Suitably, R2 is selected from R9, OR9, CO2R9, NR9R 10 、NH-C(O)-R9、O-(CH2) s -NH-C(O)-R9, O-(CH2) s -C(O)-NH-R9, COR9 and C(O)NR9R 10 .

[0161] Suitably, R2 is selected from R9, OR9, CO2R9, O-(CH2) s -NH-C(O)-R9, O-(CH2) s -C(O)-NH-R9, COR9 and C(O)NR9R 10 .

[0162] Suitably, R2 is selected from R9, O-(CH2) s -NH-C(O)-R9 and O-(CH2) s -C(O)-NH-R9.

[0163] In some embodiments, R2is H.

[0164] R3

[0165] Suitably, R3is selected from H, C 1-6 alkyl and CH2Ph.

[0166] Suitably, R3is selected from H, C

[0167] More suitably, R3is selected from methyl and ethyl.

[0168] More suitably, R3is methyl.

[0169] R4

[0170] R4is selected from phenyl and heteroaryl groups optionally substituted with up to three optional substituents. Thus, any phenyl group or C 5-9 heteroaryl group selected by R4may be optionally substituted with up to three optional substituents.

[0171] Suitably, R4is selected from phenyl, pyrrolyl, N-methylpyrrolyl, furanyl, thienyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, pyridyl, benzofuranyl, benzothienyl, benzoimidazolyl, N-methylbenzoimidazolyl, benzo oxazolyl and benzothiazolyl, optionally substituted with up to three optional substituents selected from OH, C 1-6 alkyl, OC 1-6 alkyl, (CH2) j -CO2R 11 , O-(CH2) k -NR 11 R 12 , (CH2) j -NR 11 R 12 , C(=O)-NH-(CH2) k -NR 11 R 12 ; C(=O)-NH-R 24 and C(=O)-NH-(CH2) k -C(=NH)NR 11 R 12 .

[0172] Suitably, R4is selected from phenyl, pyrrolyl, N-methylpyrrolyl, furanyl, thienyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, benzofuranyl, benzothienyl, benzoimidazolyl, N-methylbenzoimidazolyl, benzo Oxyzolyl and benzothiazolyl, optionally substituted with one or two optional substituents selected from OH, C 1-6 Alkyl, OC 1-6 Alkyl, (CH2) j -CO2R 11 、O-(CH2) k -NR 11 R 12 、(CH2) j -NR 11 R 12 、C(=O)-NH-(CH2) k -NR 11 R 12 ; C(=O)-NH-R 24 and C(=O)-NH-(CH2) k -C(=NH)NR 11 R 12 .

[0173] Suitably, R4 is selected from phenyl, N-methylpyrrolyl, thienyl, N-methylimidazolyl, oxazolyl, thiazolyl, benzothiophenyl, N-methylbenzimidazolyl and benzothiazolyl, optionally substituted with one or two optional substituents selected from OH, C 1-6 Alkyl, OC 1-6 Alkyl, (CH2) j -CO2R 11 O-(CH2) k -NR 11 R 12 、(CH2) j -NR 11 R 12 、C(=O)-NH-(CH2) k -NR 11 R 12 ; C(=O)-NH-R 24 and C(=O)-NH-(CH2) k -C(=NH)UR 11 R 12 .

[0174] Suitably, R4 is optionally substituted with up to three optional substituents selected from OH, C 1-6 Alkyl, OC 1-6 Alkyl, (CH2) j -CO2R 11 、O-(CH2) k -NH2, (CH2) j-NH2, C(=O)-NH-(CH2) k -NH2; C(=O)-NH-R 24 and C(=O)-NH-(CH2) k -C(=NH)NH2.

[0175] Suitably, R4 is optionally substituted C(=O)-NH-R 24 , where R 24 is -C6H4-(CH2) j -R 18 , and the phenylene group -C6H4- is para-substituted.

[0176] Suitably, R4 is optionally substituted with up to three optional substituents selected from OH, methyl, ethyl, OCH3, OCH2CH3, CO2H, CO2CH3, CO2CH2CH3, O-(CH2) k -NH2 and (CH2) j -NH2.

[0177] Suitably, R4 is optionally substituted with one or two optional substituents.

[0178] More suitably, R4 is optionally substituted with one optional substituent.

[0179] More suitably, R4 is selected from:

[0180]

[0181] wherein Z1 is selected from NH, N-CH3, S and O;

[0182] Z2 is selected from CH and N;

[0183] Z3 is selected from S and O;

[0184] Z4 is selected from CH and N;

[0185] R 22 Selected from (CH2) j CO2R 11 、(CH2) j NR 11 R 12 and C(=O)-NH-C6H4-(CH2) j -R 18 ;

[0186] R 18 Selected from CO2R 11 and NR 11 R 12 ;

[0187] j is selected from an integer from 0 to 6;

[0188] R 11 and R 12 are independently selected from H and C 1-6 alkyl; and

[0189] R 23 is selected from H and C 1-6 alkyl.

[0190] the wavy line indicates the point of attachment of the R4group described above to the remainder of the compound of formula (I).

[0191] More suitably, R4is selected from:

[0192]

[0193] wherein Z1is selected from NH, N-CH3, S and O;

[0194] Z2is selected from CH and N; and

[0195] Z3is selected from S and O;

[0196] Z4is selected from CH and N;

[0197] R 11 is selected from H and C 1-6 alkyl; and

[0198] R 23 is selected from H and C 1-6 alkyl.

[0199] R5 and R6

[0200] Suitably, for (iii), R5is H and R6is OC 1-6 alkyl.

[0201] Most suitably, (i) R5and R6together form a double bond.

[0202] R7

[0203] Suitably, R7is selected from H, C 1-12 alkyl, C 5-9 heteroaryl, C 6-15 heteroarylalkyl, phenyl, benzyl and phenethyl; wherein the heteroaryl, heteroarylalkyl, phenyl and aralkyl groups are optionally substituted with up to three groups selected from C 1-6 alkyl, OH, OC 1-6 alkyl.

[0204] Suitably, R7is selected from H, C 1-12Alkyl, pyrrolyl, N-methylpyrrolyl, furyl, thienyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, pyridyl, indolyl, N-methylindolyl, benzofuranyl, benzothiophenyl, benzimidazolyl, N-methylbenzimidazolyl, benzo oxazolyl, benzothiazolyl, pyrrol-3-ylmethyl, pyrrol-4-ylmethyl, imidazol-2-ylmethyl, imidazol-4-ylmethyl, thien-3-ylmethyl, furan-3-ylmethyl, phenyl, benzyl and phenethyl; wherein the heteroaryl, heteroarylalkyl, phenyl and aralkyl groups are optionally substituted with a group selected from C 1-6 Alkyl, OH, OC 1-6 The alkyl group is substituted with up to three groups.

[0205] Suitably, R7 is selected from H, C 1-6 Alkyl, pyrrolyl, N-methylpyrrolyl, furyl, thienyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, pyridyl, indolyl, N-methylindolyl, benzofuranyl, benzothiophenyl, benzimidazolyl, N-methylbenzimidazolyl, benzo oxazolyl, benzothiazolyl, pyrrol-3-ylmethyl, pyrrol-4-ylmethyl, imidazol-2-ylmethyl, imidazol-4-ylmethyl, thien-3-ylmethyl, furan-3-ylmethyl, phenyl, benzyl and phenethyl; wherein the heteroaryl, heteroarylalkyl, phenyl and aralkyl groups are optionally substituted with a group selected from C 1-6 Alkyl, OH, OC 1-6 The alkyl group is substituted with up to three groups.

[0206] Suitably, R7 is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pyrrolyl, N-methylpyrrolyl, furanyl, thienyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, pyridyl, indolyl, N-methylindolyl, benzofuranyl, benzothiophenyl, benzimidazolyl, N-methylbenzimidazolyl, benzo oxazolyl, benzothiazolyl, pyrrol-3-ylmethyl, pyrrol-4-ylmethyl, imidazol-2-ylmethyl, imidazol-4-ylmethyl, thien-3-ylmethyl, furan-3-ylmethyl, phenyl, benzyl and phenethyl, optionally with a phenyl group selected from C 1-6 Alkyl, OH, OC 1-6 The alkyl group is substituted with up to three groups.

[0207] Suitably, R7 is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pyrrolyl, N-methylpyrrolyl, furanyl, thienyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, pyridyl, indolyl, N-methylindolyl, benzofuranyl, benzothiophenyl, benzimidazolyl, N-methylbenzimidazolyl, benzo Oxyzolyl, benzothiazolyl, phenyl, benzyl and phenethyl, which are optionally substituted with a radical selected from C 1-6 Alkyl, OH, OC 1-6 The alkyl group is substituted with up to three groups.

[0208] In some embodiments, R7 is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl.

[0209] R9

[0210] Suitably, R9 is selected from H, C 1-12 Alkyl, C 5-9 Heteroaryl, C 6-15 heteroarylalkyl, phenyl, benzyl and phenethyl; wherein the heteroaryl, heteroarylalkyl, phenyl and aralkyl groups are optionally substituted with a group selected from C 1-6 Alkyl, OH, OC 1-6 The alkyl group is substituted with up to three groups.

[0211] Suitably, R9 is selected from H, C 1-12 Alkyl, pyrrolyl, N-methylpyrrolyl, furyl, thienyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, pyridyl, indolyl, N-methylindolyl, benzofuranyl, benzothiophenyl, benzimidazolyl, N-methylbenzimidazolyl, benzo oxazolyl, benzothiazolyl, pyrrol-3-ylmethyl, pyrrol-4-ylmethyl, imidazol-2-ylmethyl, imidazol-4-ylmethyl, thien-3-ylmethyl, furan-3-ylmethyl, phenyl, benzyl and phenethyl; wherein the heteroaryl, heteroarylalkyl, phenyl and aralkyl groups are optionally substituted with a group selected from C 1-6 Alkyl, OH, OC 1-6 The alkyl group is substituted with up to three groups.

[0212] Suitably, R9 is selected from H, C 1-6 Alkyl, pyrrolyl, N-methylpyrrolyl, furyl, thienyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, pyridyl, indolyl, N-methylindolyl, benzofuranyl, benzothiophenyl, benzimidazolyl, N-methylbenzimidazolyl, benzo oxazolyl, benzothiazolyl, pyrrol-3-ylmethyl, pyrrol-4-ylmethyl, imidazol-2-ylmethyl, imidazol-4-ylmethyl, thien-3-ylmethyl, furan-3-ylmethyl, phenyl, benzyl and phenethyl; wherein the heteroaryl, heteroarylalkyl, phenyl and aralkyl groups are optionally substituted with a group selected from C 1-6 Alkyl, OH, OC 1-6 The alkyl group is substituted with up to three groups.

[0213] Suitably, R9 is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pyrrolyl, N-methylpyrrolyl, furanyl, thienyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, pyridyl, indolyl, N-methylindolyl, benzofuranyl, benzothiophenyl, benzimidazolyl, N-methylbenzimidazolyl, benzo oxazolyl, benzothiazolyl, pyrrol-3-ylmethyl, pyrrol-4-ylmethyl, imidazol-2-ylmethyl, imidazol-4-ylmethyl, thien-3-ylmethyl, furan-3-ylmethyl, phenyl, benzyl and phenethyl, optionally with a phenyl group selected from C 1-6 Alkyl, OH, OC 1-6 The alkyl group is substituted with up to three groups.

[0214] Suitably, R9 is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pyrrolyl, N-methylpyrrolyl, furanyl, thienyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, pyridyl, indolyl, N-methylindolyl, benzofuranyl, benzothiophenyl, benzimidazolyl, N-methylbenzimidazolyl, benzo Oxyzolyl, benzothiazolyl, phenyl, benzyl and phenethyl, which are optionally substituted with a radical selected from C 1-6 Alkyl, OH, OC 1-6 The alkyl group is substituted with up to three groups.

[0215] In some embodiments, R9 is selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl.

[0216] R8, R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 20 and R 21

[0217] Suitably, R8, R 10 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 20 and R 21Each of the groups is independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl.

[0218] Suitably, R8, R 10 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 20 and R 21 Each of is independently selected from H, methyl and ethyl.

[0219] Suitably, R8 is H.

[0220] Suitably, R 10 It’s H.

[0221] Suitably, R 11 are each independently selected from H and methyl.

[0222] Suitably, R 12 are each independently selected from H and methyl; more suitably, R 12 Each is H.

[0223] Suitably, R 13 It’s H.

[0224] Suitably, R 14 It’s H.

[0225] Suitably, R 15 It’s H.

[0226] Suitably, R 16 It’s H.

[0227] Suitably, R 17 It's methyl.

[0228] Suitably, R 20 It’s H.

[0229] Suitably, R 21 It’s H.

[0230] R 18

[0231] Suitably, R 18 Selected from CO2H, CO2CH3, CO2CH2CH3, NH(CH3) and NH2.

[0232] R 19

[0233] Suitably, R 19 Selected from H, (CH2)t -N(CH2CH3)2、(CH2) t -N(CH3)2, (CH2) t -NH(CH2CH3),(CH2) t -NH(CH3) and (CH2) t -NH2.

[0234] More appropriately, R 19 Selected from H and (CH2) t -NH2.

[0235] R 24

[0236] Suitably, R 24 is phenyl optionally substituted with up to three optional substituents selected from OH, methyl, ethyl, propyl, OCH3, OCH2CH3, CO2H, CO2CH3, CO2CH2CH3, O-(CH2) k -NH2, O-(CH2) k -NH(CH3),(CH2) j -NH2, (CH2) j -NH(CH3), C(=O)-NH-(CH2) k -NH2, C(=O)-NH-(CH2) k -NH(CH3), C(=O)-NH-(CH2) k -C(=NH)NH(CH3) and C(=O)-NH-(CH2) k -C(=NH)NH2.

[0237] Suitably, R 24 is phenyl optionally substituted with up to three optional substituents selected from OH, methyl, ethyl, OCH3, OCH2CH3, CO2H, CO2CH3, CO2CH2CH3, O-(CH2) k -NH2 and (CH2) j -NH2.

[0238] Suitably, R 24 It is a para-substituted phenyl group.

[0239] More preferably, in some aspects, R 24 is -C6H4-(CH2) j -R 18 , where R 18 Selected from CO2R 11 and NR 11 R 12 .

[0240] j

[0241] Each instance of j is independently selected from an integer from 0 to 6, thus, each j is independently selected from 0, 1, 2, 3, 4, 5, and 6.

[0242] Suitably, each j is independently selected from 0, 1, 2 and 3.

[0243] More suitably, j is each independently selected from 0 and 1.

[0244] More suitably, j is each 0.

[0245] k

[0246] Each instance of k is independently selected from an integer from 1 to 6, thus, each k is independently selected from 1, 2, 3, 4, 5, and 6.

[0247] Suitably, k is each independently selected from 1, 2 and 3.

[0248] More suitably, k is each 1.

[0249] m

[0250] m is selected from integers from 0 to 6, and thus, m is selected from 0, 1, 2, 3, 4, 5 and 6.

[0251] Suitably, m is selected from 0, 1, 2 and 3.

[0252] More suitably, m is selected from 0 and 1.

[0253] More suitably, m is 0.

[0254] n

[0255] n is selected from integers from 1 to 6, thus, n is selected from 1, 2, 3, 4, 5 and 6.

[0256] Suitably, n is selected from 1, 2 and 3.

[0257] More suitably, n is 1.

[0258] r

[0259] r is selected from an integer from 0 to 6, thus, r is selected from 0, 1, 2, 3, 4, 5 and 6.

[0260] Suitably, r is selected from 0, 1, 2 and 3.

[0261] More suitably, r is selected from 0 and 1.

[0262] More suitably, r is zero.

[0263] s

[0264] s is selected from integers from 1 to 6, thus, s is selected from 1, 2, 3, 4, 5 and 6.

[0265] Suitably, s is selected from 1, 2 and 3.

[0266] More suitably, s is 1.

[0267] t

[0268] t is selected from integers from 0 to 6, thus, t is selected from 0, 1, 2, 3, 4, 5 and 6.

[0269] Suitably, t is selected from 0, 1, 2 and 3.

[0270] More suitably, t is selected from 0 and 1.

[0271] More suitably, t is 0.

[0272] Y1

[0273] Y1 is N or CH; suitably, Y1 is CH.

[0274] Y2

[0275] Y2 is N or CH; suitably, Y2 is CH.

[0276] X1

[0277] Suitably, X1 is selected from O, S, NH, CH2, CH2O, C(=O), C(=O)NR 13 NR 13 C(=O), OC(O) and C(O)-O;

[0278] Suitably, X1 is selected from O, C(=O), C(=O)NR 13 and NR 13 C(=O).

[0279] More suitably, X1 is selected from O, C(=O)NH and NHC(=O).

[0280] More suitably, X1 is O.

[0281] X2

[0282] Suitably, X2 is selected from O, S, NH, CH2, CH2O, C(=O), C(=O)NR 15 NR 15C(=O), OC(O) and C(O)-O may not exist.

[0283] Suitably, X2 is selected from O, C(=O), C(=O)NR 15 and NR 16 C(=O) or does not exist.

[0284] More suitably, X2 is selected from O, C(=O)NH and NHC(=O).

[0285] Suitably, X2 is the same as X1.

[0286] More suitably, X2 is O.

[0287] L

[0288] L is a linker group. Suitably, any of the peptide chain, alkylene chain, paraformaldehyde chain or polyethylene glycol chain is replaced by one or more heteroatoms (e.g., N, O and S) and / or one or more C 5-9 The chain may be interrupted by one to three heteroatoms and / or one to three C 5-9 The phenylene group is interrupted by a heteroarylene group and / or one to three phenylene groups.

[0289] Suitably, L is selected from a peptide chain having 2 to 5 amino acids, 2 to 4 amino acids, 2 to 3 amino acids; an alkylene chain containing 1 to 11 carbon atoms, 1 to 10 carbon atoms, 1 to 9 carbon atoms, 1 to 8 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, which may contain one or more carbon-carbon double bonds or triple bonds; a paraformaldehyde chain -(OCH2) 1-12 -、-(OCH2) 1-11 -、-(OCH2) 1-10 -、-(OCH2) 1-9 -、-(OCH2) 1-8 -、-(OCH2) 1-7 -、-(OCH2) 1-6 -、-(OCH2) 1-5 -、-(OCH2) 1-4 -、-(OCH2) 1-3 -, polyethylene glycol chain -(OCH2CH2) 1-5 -, chain-(OCH2CH2) 1-4 -, chain-(OCH2CH2) 1-3 -; the chain may be separated by one or more heteroatoms and / or C5-9 The phenylene group is interrupted by a heteroarylene group and / or one to three phenylene groups.

[0290] More suitably, L may be selected from a hydrocarbylene chain containing from 1 to 12 carbon atoms, which may contain one or more carbon-carbon double or triple bonds.

[0291] More suitably, L may be selected from CH═CH, CH2, CH2CH2, CH2CH2CH2, CH2CH2CH2CH2 and CH2CH2CH2CH2CH2CH2.

[0292] A

[0293] In one embodiment, A is A1:

[0294]

[0295] wherein for each A1 group, one of Y3 and Y4 is independently selected from NR 17 , S and O; and the other of Y3 and Y4 is CH; and Y5 is independently selected from CH, N, S and COH.

[0296] In this embodiment, when q is selected from 2, 3, 4, 5 and 6, then A will contain a plurality of A1 groups linked to each other.

[0297] Thus, the 5-membered ring containing Y3, Y4 and Y5 is a heteroaromatic ring. The A1 group can be connected to the rest of the molecule in any direction. Thus, when A is A1, as in the above embodiment, the compound of formula (I) is selected from:

[0298]

[0299] More suitably, when A is A1, the compound of formula (I) is compound (II).

[0300] Therefore, the heteroaromatic ring containing Y3, Y4 and Y5 is selected from one of the following groups:

[0301]

[0302]

[0303] More appropriately, A is

[0304]

[0305] wherein Y5 is selected from CH and N.

[0306] In another embodiment, A is A2:

[0307]

[0308] wherein for each A2 group, one of Y6 and Y7 is independently selected from N and CH; and the other of Y6 and Y7 is CH.

[0309] Thus, the 6-membered ring containing Y6 and Y7 is a phenyl or pyridine ring. The A2 group can be connected to the rest of the molecule in any direction. Thus, when A is A2, as in the above embodiment, the compound of formula (I) is selected from:

[0310]

[0311] More suitably, when A is A2, the compound of formula (I) is compound (IV).

[0312] Suitably, A is A4:

[0313]

[0314] More suitably, Y6 is CH; and Y7 is CH.

[0315] q

[0316] Suitably, q is selected from 0, 1, 2 and 3.

[0317] More suitably, q is 0 or 1.

[0318] 6-membered aromatic ring

[0319] Suitably, the 6-membered aromatic ring of formula (I) is para-substituted:

[0320]

[0321] More suitably, the 6-membered aromatic ring of formula (I) is:

[0322]

[0323] Optional double bond in the C-ring

[0324] The present invention provides compounds of formula (I):

[0325]

[0326] The dashed lines indicate the optional presence of double bonds between one or more of C1 and C2, C2 and C3, and C3 and C4.

[0327] In one aspect, the compound of formula (I) has a double bond between C1 and C2 to give a compound of formula (VI):

[0328]

[0329] In another aspect, the compound of formula (I) has a double bond between C2 and C3 to give a compound of formula (VII):

[0330]

[0331] In another aspect, the compound of formula (I) has a double bond between C3 and C4 to give a compound of formula (VIII):

[0332]

[0333] In another aspect, the compound of formula (I) has a double bond between C1 and C2 and a double bond between C3 and C4 to give a compound of formula (IX):

[0334]

[0335] Other restrictions

[0336] Options for compounds of formula (I) include the following: when p is 0 and A is A1, then: (a) for at least one A1 group, one of Y3 and Y4 is selected from S and O; or (b) for at least one A1 group, Y5 is S; or (c) R4 is not optionally substituted pyrrolyl or imidazolyl.

[0337] When q is selected from 2, 3, 4, 5 and 6, more than one A1 group will be present.

[0338] Thus, when p is 0 and A is A1, the condition requires the presence of at least one aryl group, or, alternatively, the presence of a heteroaryl group (or as part of A1 or R4) that does not contain a 5-membered pyrrole or imidazole ring, or an optionally substituted derivative such as N-methylpyrrole or N-methylimidazole ring. Thus, this condition avoids the compound of formula (I) having only poly-pyrrole or poly-imidazole or poly-pyrrole-imidazole long chain groups attached to PDD. Compounds with such long chain groups tend to have relatively poor cytotoxicity.

[0339] In some aspects, suitably, the options for compounds of formula (I) contain the following proviso: when p is 0 and A is A1, then: (a) the 5-membered ring of A1 is selected from H9, H10, H11, H12, H13, H14, H15, H16, H17, H19, H20, H21, H22, H23 and H24; or (b) the 5-membered ring of A1 is selected from H5 and H6; or (c) R4 is selected from phenyl, furyl, thiophenyl, oxazolyl, thiazolyl, pyridyl, benzofuranyl, benzothiophenyl, benzimidazolyl, N-methylbenzimidazolyl, benzo oxazolyl and benzothiazolyl, optionally substituted with up to three optional substituents selected from OH, C 1-6 Alkyl, OC 1-6 Alkyl, (CH2) j -CO2R 11 、O-(CH2) k -NR 11 R 12 、(CH2) j -NR 11 R 12 、C(=O)-NH-(CH2) k -NR 11 R 12 ; C(=O)-NH-C6H4-(CH2) j -R 18 and C(=O)-NH-(CH2) k -C(=NH)NR 11 R 12 .

[0340] In some aspects, suitably, the options for compounds of formula (I) contain the proviso that when p is 0 and A is A1, then: (a) the 5-membered ring of A1 is selected from H9, H10, H11, H12, H13, H14, H15, H16, H17, H19, H20, H21, H22, H23 and H24; or (b) the 5-membered ring of A1 is selected from H5 and H6; or (c) R4 is selected from phenyl and C9 heteroaryl groups optionally substituted with up to three optional substituents selected from OH, C 1-6 Alkyl, OC 1-6 Alkyl, (CH2) j -CO2R 11 、O-(CH2) k -NR 11 R 12 、(CH2) j -NR 11 R 12 、C(=O)-NH-(CH2) k -NR 11 R 12 ; C(=O)-NH-C6H4-(CH2) j -R 18 ; C(=O)-NH-(CH2) k -C(=NH)NR 11 R 12 , the condition is C 5-9Heteroaryl is not indolyl.

[0341] Suitable structure

[0342] Draw the compound of formula (I) without specifying the position of R1 and R2 on the C-ring:

[0343]

[0344] Thus, R1 and R2 may be present at any position of the C-ring, provided that the valence requirements are met. Since the fused carbon and nitrogen of the C-ring all have the substituents shown, this means that R1 and R2 may be present at any non-fused carbon of the C-ring (i.e., the C1, C2, C3, or C4 positions specified above). Suitably, R1 and R2 are present at two different non-fused carbons of the C-ring.

[0345] In one aspect, the compound of formula (I) is selected from:

[0346]

[0347] In another aspect, the compound of formula (I) is selected from:

[0348]

[0349] More suitably, the compound of formula (I) has the following structure:

[0350]

[0351]

[0352] For compounds of formula (XV) wherein R1 and / or R2 are substituents other than H, the carbon in the C-ring to which any such substituent is attached will be a stereocenter. In formula (XV), R1 and R2 are drawn without specifying the stereochemistry of the carbons on the C-ring to which they are attached.

[0353] More suitably, the compound of formula (I) is selected from:

[0354]

[0355] wherein q is selected from 0, 1, 2, 3, 4, 5 or 6;

[0356] p is 0 or 1;

[0357] L is an alkylene chain containing 1 to 12 carbon atoms;

[0358] Y1 is N or CH;

[0359] Y2 is N or CH; and wherein at least one of Y1 and Y2 is CH;

[0360] Y5 is selected from CH and N;

[0361] Z1 is selected from O, S, NH and N-CH3;

[0362] Z2 is selected from CH and N;

[0363] Z3 is selected from S and O;

[0364] Z4 is selected from CH and N;

[0365] R 22 Selected from (CH2) j CO2H, (CH2) j CO2C 1-6 Alkyl, (CH2) j NR 11 R 12 and C(=O)-NH-C6H4-(CH2) j -R 18 ;

[0366] R 18 Selected from CO2R 11 and NR 11 R 12 ;

[0367] R 19 Selected from H and (CH2) t -NR 20 R 21 ;

[0368] j and t are independently selected from integers from 0 to 6; and

[0369] R 11 、R 12 and R 23 Independently selected from H and C 1-6 alkyl.

[0370] or:

[0371] (i) R5 and R6 together form a double bond;

[0372] (ii) R5 is H and R6 is OH; or

[0373] (iii) R5 is H and R6 is OC 1-6 alkyl;

[0374] Provided that when the compound is (XVI) and p is 0, then Z1 is selected from O and S.

[0375] More suitably, the compound of formula (I) is selected from:

[0376]

[0377] wherein q is selected from 0, 1, 2, 3, 4, 5 or 6;

[0378] p is 0 or 1 ;

[0379] L is a hydrocarbylene chain containing 1 to 12 carbon atoms;

[0380] Y1is N or CH;

[0381] Y2is N or CH; and wherein at least one of Y1and Y2is CH;

[0382] Y5is selected from CH and N;

[0383] Z1is selected from O, S, NH and N-CH3;

[0384] Z2is selected from CH and N;

[0385] Z3is selected from S and O;

[0386] Z4is selected from CH and N;

[0387] R 19 is selected from H and (CH2) t -NR 20 R 21 ;

[0388] t is selected from an integer from 0 to 6;

[0389] R 11 , R 20 , R 21 and R 23 are independently selected from H and C 1-6 alkyl;

[0390] or:

[0391] (i) R5and R6together form a double bond;

[0392] (ii) R5is H and R6is OH; or

[0393] (iii) R5is H and R6is OC 1-6 alkyl;

[0394] with the proviso that when the compound is (XX) and p is 0, then Z1is selected from O and S.

[0395] More suitably, the compound of formula (I) is selected from:

[0396] (a) (S)-5-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido-[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-pyrrole-2-carboxamido)benzo-[b]thiophene-2-carboxylic acid methyl ester (13)

[0397]

[0398] (b) (S)-5-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido-[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-imidazole-2-carboxamido)-benzo[b]thiophene-2-carboxylic acid methyl ester (17)

[0399]

[0400] (c) (S)-4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]-pyrido[1,2-a][1,4]diazepine (3-amino-1-methyl-1H-imidazole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylic acid methyl ester (20)

[0401]

[0402] (d) (S)-4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]-pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylic acid methyl ester (24)

[0403]

[0404] (e)(S)-4-(4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]-pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-benzamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylic acid methyl ester (28)

[0405]

[0406] (f) (S)-5-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido-[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-benzamido)benzo[b]thiophene-2-carboxylic acid methyl ester (30)

[0407]

[0408] (g) (S)-4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido-[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-imidazole-2-carboxamido)-benzoic acid methyl ester (34)

[0409]

[0410] (h)(S)-4-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido-[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-pyrrole-2-carboxamido)-benzoate methyl ester (38)

[0411]

[0412] (i) (S)-N-(4-aminophenyl)-4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydro-benzo[e]pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyl-amido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxamide (41)

[0413]

[0414] (j) (S)-N-(2-((4-aminophenyl)carbamoyl)benzo[b]thiophen-5-yl)-4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine) -3-yl)oxy)-butyramido)-1-methyl-1H-pyrrole-2-carboxamide (47)

[0415]

[0416] (k) (S)-N-(4-aminophenyl)-4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -3-yl)oxy)hexanoyl)-1-methyl-1H-pyrrole-2-carboxamide)phenyl)-1-methyl-1H-pyrrole-2-carboxamide (62)

[0417]

[0418]

[0419] (1) (S)-N-(4-aminophenyl)-4-(6-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -3-yl)oxy)hexane-amido)-1-methyl-1H-pyrrole-2-carboxamide (66)

[0420]

[0421] and

[0422] (m) (S)-N-(2-aminoethyl)-4-(4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydro-benzo[e]pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxamide (68)

[0423]

[0424] In a further aspect, there is provided a compound of formula (I):

[0425]

[0426] and salts and solvates thereof, wherein:

[0427] Dashed lines indicate the optional presence of double bonds between one or more of C1 and C2, C2 and C3, and C3 and C4;

[0428] R1 is selected from R7, =CH2, =CH-(CH2) m -CH3, =O, (CH2) m -OR7, (CH2) m-CO2R7, (CH2) m -NR7R8, O-(CH2) n -NR7R8, NH-C(O)-R7, O-(CH2) n -NH-C(O)-R7, O-(CH2) n -C(O)-NH-R7, (CH2) m -SO2R7, O-SO2R7, (CH2) m -C(O)R7 and (CH2) m -C(O)NR7R8;

[0429] R2 is selected from R9, =CH2, =CH-(CH2) r -CH3, =O, (CH2) r -OR9, (CH2) r -CO2R9, (CH2) r -NR9R 10 、O-(CH2) s -NR9R 10 、NH-C(O)-R9、O-(CH2) s -NH-C(O)-R9, O-(CH2) s -C(O)-NH-R9, (CH2) r -SO2R9, O-SO2R9, (CH2) r -COR9 and (CH2) r -C(O)NR9R 10 ;

[0430] R3 is selected from H, C 1-12 Alkyl and CH2Ph;

[0431] R4 is selected from phenyl and C 5-9 Heteroaryl group, the substituent is selected from OH, C 1-6 Alkyl, OC 1-6 Alkyl, (CH2) j -CO2R 11 、O-(CH2) k -NR 11 R 12 、(CH2) j -NR 11 R 12 、C(=O)-NH-(CH2) k -NR 11 R 12 ; C(=O)-NH-C6H4-(CH2) j -R 18and C(=O)-NH-(CH2) k -C(=NH)NR 11 R 12 ;

[0432] R 19 Selected from H and (CH2) t -NR 20 R 21 ;

[0433] Y1 is N or CH; Y2 is N or CH; and wherein at least one of Y1 and Y2 is CH;

[0434] p is 0 or 1; j, m, r and t are independently selected from integers from 0 to 6;

[0435] k, n and s are independently selected from integers from 1 to 6;

[0436] X1 is selected from O, S, NR 13 , CR 13 R 14 , CR 13 R 14 O, C(=O), C(=O)NR 13 NR 13 C(=O), OC(O) and C(O)-O;

[0437] L is selected from amino acids, peptide chains having 2 to 6 amino acids, alkylene chains containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, paraformaldehyde chains -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 -, the chain may be substituted by O, S and / or NH groups and / or C 3-9 interrupted by one or more of heteroarylene and / or phenylene;

[0438] X2 is selected from O, S, NR 15 , CR 15 R 16 , CR 15 R 16 O, C(=O), C(=O)NR 15 NR 15 C(=O), OC(O) and C(O)-O or absent;

[0439] q is selected from 0, 1, 2, 3, 4, 5 and 6;

[0440] A is selected from:

[0441]

[0442] For each A1 group, one of Y3 and Y4 is independently selected from NR 17 , S and O; and the other of Y3 and Y4 is CH; and Y5 is independently selected from CH, N, S and COH; and

[0443] For each A2 group, one of Y6 and Y7 is independently selected from N and CH; and the other of Y6 and Y7 is CH;

[0444] R7 and R9 are independently selected from H, C 1-12 Alkyl, C 5-9 Heteroaryl, C 6-15 Heteroarylalkyl, phenyl and C 7-12 Aralkyl groups; wherein the heteroaryl, heteroarylalkyl, phenyl and aralkyl groups are optionally substituted with up to three optional substituents selected from C 1-6 Alkyl, OH, OC 1-6 alkyl;

[0445] R 18 Selected from CO2R 11 and NR 11 R 12 ;

[0446] R8, R 10 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 20 and R 21 Independently selected from H and C 1-6 alkyl; and

[0447] (i) R5 and R6 together form a double bond; or (ii) R5 is H and R6 is OH; or (iii) R5 is H and R6 is OC 1-6 alkyl.

[0448] application

[0449] The present invention can be applied to the treatment of proliferative diseases.

[0450] In certain aspects, methods of treating a proliferative disease are provided, comprising administering to a subject a therapeutically effective amount of a compound of Formula (I), or salts and solvates thereof, or a composition comprising a compound of Formula (I), or salts and solvates thereof.

[0451] In certain aspects, methods of treating a proliferative disease are provided, the methods comprising administering to a subject a therapeutically effective amount of a targeted conjugate comprising a compound of Formula (I) and salts and solvates thereof.

[0452] In certain aspects, methods of treating a proliferative disease are provided, the methods comprising administering to a subject a therapeutically effective amount of an antibody-drug conjugate comprising a compound of Formula (I) and salts and solvates thereof.

[0453] The term "proliferative disorder" refers to excessive or abnormal unwanted or uncontrolled cell proliferation, whether in vitro or in vivo, of cells that is undesirable such as neoplastic or hyperplastic growth. Examples of proliferative conditions include, but are not limited to, benign, premalignant and malignant cell proliferations, including, but not limited to, neoplasms and tumors (e.g., histiocytomas, gliomas, astrocytomas, osteomas), cancers (e.g., lung cancer, small cell lung cancer, hepatocellular carcinoma, gastric or stomach cancer (including gastrointestinal cancer), intestinal cancer, colon cancer, liver cancer, breast cancer, glioblastoma, cervical cancer, ovarian cancer, esophageal cancer [or esophageal cancer], oral cancer, prostate cancer, testicular cancer, liver cancer, rectal cancer, colorectal cancer, endometrial or uterine cancer, uterine cancer, salivary gland cancer, renal or kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penile cancer, head and neck cancer, bladder cancer, pancreatic cancer, brain cancer, sarcomas, osteosarcomas, Kaposi's sarcoma, melanoma), leukemias, psoriasis, bone diseases, fibroproliferative disorders (e.g., fibroproliferative disorders of connective tissue), and atherosclerosis. Suitably, the proliferative disease is selected from bladder cancer, bone cancer, intestinal cancer, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, esophageal cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, retinoblastoma, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer and uterine cancer. Suitably, the proliferative disease is selected from breast cancer and cervical cancer.

[0454] Any cell type can be treated, including but not limited to bone, eye, head and neck, lung, gastrointestinal (including, for example, oral, esophageal, intestinal, colon), breast (breast), cervix, ovary, uterine, prostate, liver (hepatocytes), kidney (renal cells), bladder, pancreas, brain, and skin.

[0455] A skilled artisan can readily determine whether a candidate compound treats a proliferative condition of any particular cell type.

[0456] Suitably, the subjects are humans, livestock animals and companion animals.

[0457] In other aspects, the compound of formula (I) and its salt and solvate can be directly or indirectly connected to a targeting agent (e.g., an antibody, an antibody fragment, a hormone, etc.) to provide a targeted conjugate. The targeted conjugate of the present disclosure can contain one or more compounds of formula (I) (or its salt and solvate). A variety of targeted conjugates are known in the art and can be used together with the compound of formula (I) and its salt and solvate. For example, in a particular aspect, the targeted conjugate is an antibody-drug conjugate, wherein one or more compounds of formula (I) are directly or indirectly connected to an antibody. Therefore, the compound of formula (I) and its salt and solvate can be used as an effective load on the targeted conjugate.

[0458] Suitably, the compound of formula (I) and its salt and solvate are used as drugs in targeted conjugates, which are prepared by connecting the compound of formula (I) and its salt and solvate directly or via an optional linker group to a targeting agent. Suitably, the compound of formula (I) and its salt and solvate are connected to the targeting agent via a linker group. Suitably, the targeted conjugate is used in the treatment of diseases, more particularly in the treatment of proliferative diseases. Suitably, the drug can be directly or via a linker group connected to the targeting agent by any suitable functional group it contains. Typically, the drug contains or can be modified to contain one or more functional groups, such as amine, hydroxyl or carboxylic acid groups, for connecting the drug directly or via a linker group to the targeting agent. In some aspects, one or more atoms or groups of the compound of formula (I) can be eliminated during the drug connection to the antibody. In some aspects, the targeting agent is combined with a cell surface receptor or a tumor-associated antigen. In some aspects, the targeting agent is an antibody. In some aspects, the targeting agent is a hormone. In some aspects, the targeting agent is a protein. In some aspects, the targeting agent is a polypeptide. In some aspects, the targeting agent is a small molecule (eg, folic acid).

[0459] The compounds of formula (I) can be applied as payloads for antibodies or antibody fragments. The compounds of formula (I) readily allow conjugation to antibodies or antibody fragments.

[0460] Antibody-drug conjugates

[0461] Antibody therapy has been established for the targeted treatment of patients with cancer, immune and angiogenic disorders (Carter, P. (2006) Nature Reviews Immunology 6: 343-357). The use of antibody-drug conjugates (ADCs) (i.e., immunoconjugates) for local delivery of cytotoxic or cytostatic agents (i.e., drugs that kill or inhibit tumor cells in the treatment of cancer) targets the delivery of the drug moiety to the tumor and its intracellular accumulation therein, whereas systemic administration of these unconjugated agents can produce unacceptable levels of toxicity to cells (Xie et al. (2006) Expert. Opin. Biol. Ther. 6(3):281-291; Kovtun ef a / (2006) Cancer Res. 66(6):3214-3121; Law et al. (2006) Cancer Res. 66(4):2328-2337; Wu et al. (2005) Nature Biotech. 23(9):1137-1145; Lambert J. (2005) Current Opin. in Pharmacol. 5:543-549; Hamann et al. (2006) Cancer Res. 66(7):1140-1157; Lambert J. (2005) Current Opin. in Pharmacol. 5:543-549). P. (2005) Expert Opin. Ther. Patents 15(9): 1087-1103; Payne, G. (2003) Cancer Cell 3: 207-212; Trailef a / (2003) Cancer Immunol. Immunother. 52: 328-337; Syrigos and Epenetos (1999) Anticancer Research 19:605-614).

[0462] Thus, maximum efficacy with minimal toxicity is sought. Efforts to design and improve ADCs have focused on the selectivity of monoclonal antibodies (mAbs) as well as the drug action mechanism, drug attachment, drug / antibody ratio (loading), and drug release characteristics (Junutula et al., 2008b Nature Biotech., 26(8):925-932; Doman et al., (2009) Blood 114(13):2721-2729; US 7521541; US 7723485; WO2009 / 052249; McDonagh (2006) Protein Eng. Design & Sel. 19(7): 299-307; Doronina et al., (2006) Bioconj. Chem. 17: 114-124; Erickson et al., (2006) Cancer Res. 66(8): 1-8; et al., (2005) Clin. Cancer Res. 11: 843-852; Jeffrey et al., (2005) J. Med. Chem. 48: 1344-1358; Hamblett et al., (2004) Clin. Cancer Res. 10: 7063-7070).

[0463] In some aspects, the present invention relates to compounds of formula (I) and salts and solvates thereof, which are used for use as drugs in antibody-drug conjugates. Suitably, compounds of formula (I) and salts and solvates thereof are used for use as drugs in antibody-drug conjugates, and the conjugate is prepared by making the compound of formula (I) and salts and solvates thereof directly or via optional linker groups connected to antibodies. Suitably, compounds of formula (I) and salts and solvates thereof are connected to antibodies via linker groups. Suitably, antibody-drug conjugates are used in the treatment of diseases, more particularly for the treatment of proliferative diseases. Suitably, antibody-drug conjugates are used in the treatment of diseases, more particularly for the treatment of proliferative diseases. Suitably, the drug can be directly or via a linker group connected to the antibody by any suitable functional group it contains. Typically, the drug contains or can be modified to contain one or more functional groups, such as amine, hydroxyl or carboxylic acid groups, for making the drug directly or via a linker group connected to the antibody. In some aspects, the antibody of the antibody-drug conjugate is an antibody fragment, such as but not limited to a single-chain antibody. In some aspects, one or more atoms or groups of the compound of formula (I) may be eliminated during the attachment of the drug to the antibody. In some aspects, the antibody binds to a cell surface receptor or tumor associated antigen.

[0464] In some aspects, the present invention relates to the use of compounds of formula (I) and salts and solvates thereof as drugs in antibody-drug conjugates. Suitably, the use of compounds of formula (I) and salts and solvates thereof as drugs in antibody-drug conjugates is achieved by making the compounds of formula (I) and salts and solvates thereof directly or via optional linker groups connected to antibodies. Suitably, compounds of formula (I) and salts and solvates thereof are connected to antibodies via linker groups. Suitably, antibody-drug conjugates are used in the treatment of diseases, more particularly in the treatment of proliferative diseases. Suitably, the drug can be directly or via a linker group connected to the antibody by any suitable functional group it contains. Typically, the drug contains or can be modified to contain one or more functional groups, such as amine, hydroxyl or carboxylic acid groups, for making the drug directly or via a linker group connected to the antibody. In some aspects, the antibody of the antibody-drug conjugate is an antibody fragment, such as but not limited to a single-chain antibody. In some aspects, one or more atoms or groups of the compound of formula (I) can be eliminated during the drug connection to the antibody. In some aspects, the antibody is bound to a cell surface receptor or a tumor-associated antigen.

[0465] The substituents of the compounds of formula (I) can interact with DNA sequences and can be selected so as to target specific sequences. In particular, the following groups in the compounds of formula (I):

[0466]

[0467] can be selected to target specific sequences. Thus, when the substituents are tailored in this manner, the compounds of formula (I) may be used in targeted chemotherapy.

[0468] Antibodies and antibody fragments

[0469] The term "antibody" specifically encompasses monoclonal antibodies, polyclonal antibodies, dimers, multimers, multispecific antibodies (e.g., bispecific antibodies), complete antibodies, and antibody fragments, as long as they exhibit the desired biological activity (e.g., the ability to bind to a desired antigen on a target cell or target tissue). Antibodies can be murine, human, humanized, chimeric, or derived from other species. Antibodies are proteins produced by the immune system that are able to recognize and bind to specific antigens. (Janeway, C, Travers, P., Walport, M., Shlomchik (2001) Immuno Biology, 5th edition, Garland Publishing, New York). Target antigens typically have many binding sites, also called epitopes, which are recognized by the CDRs on the antibody. Each antibody that specifically binds to a different epitope has a different structure. Therefore, one antigen can have more than one corresponding antibody. Antibodies include full-length immunoglobulin molecules or immunologically active portions of full-length immunoglobulin molecules, i.e., molecules or portions thereof that contain an antigen binding site that immunospecifically binds to an antigen of a target of interest, including but not limited to cancer cells or cells that produce autoimmune antibodies associated with an autoimmune disease. The immunoglobulin can be of any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass or allotype (e.g., human G1 m1, G1 m2, G1 m3, non-G1 m1 [which is any allotype other than G1 m1], G1 m17, G2m23, G3m21, G3m28, G3m1 1, G3m5, G3m13, G3m14, G3m10, G3m15, G3m16, G3m6, G3m24, G3m26, G3m27, A2m1, A2m2, Km1, Km2, and Km3). The immunoglobulin can be derived from any species, including human, murine, or rabbit.

[0470] As used herein, "binds to an epitope" is used to mean that the antibody binds to the epitope with a higher affinity than a nonspecific partner such as bovine serum albumin (BSA, Genbank Accession No. CAA76847, Version No. CAA76847.1 G1: 3336842, Record Update Date: January 7, 2011, 02:30 PM). In some embodiments, the antibody binds to the epitope with an association constant of at least 2, 3, 4, 5, 10, 20, 50, 100, 200, 500, 1000, 2000, 5000, 10 ... 4 , 10 5 or 10 6-fold higher association constant (Ka) for binding to the epitope.

[0471] The term "antibody fragment" refers to a portion of a full-length antibody, such as its antigen-binding region or variable region. Examples of antibody fragments include Fab, Fab', F(ab')2, and scFv fragments; diabodies; linear antibodies; fragments produced by Fab expression libraries, anti-idiotype (anti-Id) antibodies, CDRs (complementarity determining regions), single-chain antibody molecules; and multispecific antibodies formed by any of the above antibody fragments and epitope-binding fragments that immunospecifically bind to target antigens (such as, for example, cancer cell antigens, viral antigens, or microbial antigens). As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, that is, the individual antibodies constituting the population are identical, except for possible naturally occurring mutations that may be present in trace amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. In addition, in contrast to polyclonal antibody preparations comprising different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant or epitope on the antigen. In addition to their specificity, the advantage of monoclonal antibodies is that they can be synthesized without being contaminated by other antibodies. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be prepared by the hybridoma method first described by Kohler et al. (1975) Nature 256:495, or may be prepared by recombinant DNA methods (see US 4816567). Monoclonal antibodies may also be isolated from phage antibody libraries using the techniques described in Clackson et al. (1991) Nature, 352:624-628; Marks et al. (1991) J. Mol. Biol., 222:581-597, or from transgenic mice carrying the complete human immunoglobulin system (Lonberg (2008) Curr. Opinion 20(4):450-459).

[0472] Antibodies herein, including monoclonal antibodies, specifically include "chimeric" antibodies in which a portion of the antibody (e.g., the heavy and / or light chain) is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (US 4816567; and Morrison et al. (1984) Proc. Natl. Acad. Sci. USA, 81 :6851-6855). Chimeric antibodies include "primatized" antibodies comprising variable domain antigen binding sequences derived from a non-human primate (e.g., Old World Monkey or Ape) and human constant region sequences. An "intact antibody" herein is one comprising VL and VH domains, and a light chain constant domain (CL) and heavy chain constant domains CH1, CH2 and CH3. The constant domains can be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variant of the native sequence constant domains. An intact antibody can have one or more "effector functions" which refer to those biological activities attributable to the Fc region (a native sequence Fc region or amino acid sequence variant Fc region) of an antibody. Examples of antibody effector functions include C1q binding; complement dependent cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; and down regulation of cell surface receptors (such as B cell receptor and BCR), for example.

[0473] The antibodies disclosed herein can be modified. For example, to make them less immunogenic to human subjects. This can be achieved using any of a variety of techniques familiar to those skilled in the art, such as humanization.

[0474] Administration & Dosage

[0475] The compounds of formula I can be administered alone or in combination with one or another of the compounds described, or with one or more pharmacologically active compounds different from the compounds of formula I.

[0476] The compound of the present invention can be suitably combined with various components to produce the composition of the present invention. Suitably, the composition is combined with a pharmaceutical carrier or diluent to produce a pharmaceutical composition (which can be used for human or animal use). Suitable carriers and diluents include isotonic saline solutions, such as phosphate buffered saline. Useful pharmaceutical compositions and their preparation methods can be found in standard pharmaceutical literature. See, for example, Handbook for Pharmaceutical Additives, 3rd edition (M.Ash and I.Ash ed.), 2007 (Synapse Information Resources, Inc., Endicott, NewYork, USA) and Remington: The Science and Practice of Pharmacy, 21st edition (D.B.Troy ed.) 2006 (Lippincott, Williams and Wilkins, Philadelphia, USA), which are incorporated herein by reference.

[0477] The compounds of the present invention may be administered by any suitable route. Suitably, the compounds of the present invention will generally be administered orally or by any parenteral route in the form of a pharmaceutical preparation comprising the active ingredient, optionally in the form of a non-toxic organic or inorganic acid or base or addition salt, in a pharmaceutically acceptable dosage form.

[0478] The compounds of the invention, their pharmaceutically acceptable salts and pharmaceutically acceptable solvates of either entity can be administered alone but will generally be administered in admixture with a suitable pharmaceutical excipient, diluent or carrier selected with regard to the intended route of administration and standard pharmaceutical practice.

[0479] For example, the compound of the present invention or its salt or solvate can be orally administered, buccal or sublingually in the form of tablets, capsules (including soft gel capsules), ovules, elixirs, solutions or suspensions, which may contain flavorings or coloring agents for direct-, delayed-, modified-, sustained-, controlled-release or pulse delivery applications. The compound of the present invention can also be administered in a rapidly dispersing or rapidly dissolving dosage form.

[0480] Such tablets may contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, calcium hydrogen phosphate, and glycine, disintegrants such as starch (preferably corn, potato, or tapioca starch), sodium starch glycolate, croscarmellose sodium, and certain complex silicates, and granulation binders such as polyvinylpyrrolidone, hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose (HPC), sucrose, gelatin, and acacia. In addition, lubricants such as magnesium stearate, stearic acid, glyceryl behenate, and talc may be included.

[0481] Solid compositions of similar types can also be used as fillers in gelatin capsules. In this regard, preferred excipients include lactose, starch, cellulose, lactose, or high molecular weight polyethylene glycols. For aqueous suspensions and / or elixirs, the compounds of the invention can be combined with various sweeteners or flavorings, colorants or dyes, emulsifiers and / or suspending agents, and diluents (such as water, ethanol, propylene glycol, and glycerol), and combinations thereof.

[0482] Modified release and pulse release dosage forms can contain excipients, such as those excipients described in detail for immediate release dosage forms and other excipients serving as release rate modifiers, which are coated on the device body and / or included in the device body. Release rate modifiers include, but are not limited to, hydroxypropyl methylcellulose, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, cellulose acetate, polyethylene oxide, xanthan gum, carbomer, ammonium methacrylate copolymer, hydrogenated castor oil, carnauba wax, paraffin, cellulose acetate phthalate, hydroxypropyl methylcellulose phthalate, methacrylic acid copolymer and mixtures thereof. Modified release and pulse release dosage forms can contain one or combination of release rate modifiers. Release rate modifiers can be present in the dosage form (i.e., in the matrix) and / or on the dosage form (i.e., on the surface or coating).

[0483] Fast dispersing or dissolving dosage form formulations (FDDF) may contain the following ingredients: aspartame, acesulfame potassium, citric acid, croscarmellose sodium, crospovidone, diascorbic acid, ethyl acrylate, ethyl cellulose, gelatin, hydroxypropyl methylcellulose, magnesium stearate, mannitol, methyl methacrylate, mint flavor, polyethylene glycol, fumed silica, silicon dioxide, sodium starch glycolate, sodium stearyl fumarate, sorbitol, and xylitol.

[0484] The compounds of the present invention can also be administered parenterally (e.g., intravenously, intra-arterially), or they can be administered by infusion techniques. For such parenteral administration, they are preferably used in the form of a sterile aqueous solution, which may contain other substances, such as enough salts or glucose to make the solution isotonic with the blood. If necessary, the aqueous solution should be appropriately buffered (preferably to a pH of 3-9). Suitable parenteral preparations prepared under aseptic conditions are easily completed by standard pharmaceutical techniques well known to those skilled in the art.

[0485] Suitably, the formulations of the present invention are optimized for administration routes such as oral, intravenous, etc.

[0486] During the course of treatment, administration can be carried out continuously or intermittently (e.g., in divided doses at appropriate intervals) with one dose. Methods for determining the most effective means and dosage are well known to those skilled in the art and will vary with the formulation used for treatment, the therapeutic purpose, the target cell being treated, and the subject being treated. Single or multiple administrations can be performed at the dose level and dosage regimen selected by the treating physician, veterinarian, or clinician.

[0487] Depending on the condition and patient to be treated and the route of administration, the composition may be administered at varying doses. For example, a typical dose for an adult may be 100 ng to 25 mg (suitably, about 1 μg to about 10 mg) per kg of subject body weight per day.

[0488] Suitably, when estimating initial doses for human subjects, guidance can be obtained from studies in experimental animals. For example, when a specific dose is identified for mice, suitably, the initial test dose for humans can be about 0.5 to 2 times the mg / kg value given to mice.

[0489] Other forms

[0490] Unless otherwise indicated, included in the above description are the well-known ionic, salt, solvate and protected forms of these substituents. For example, a reference to a carboxylic acid (-COOH) also includes the anionic (carboxylate) form (-COO-), its salts or solvates, and conventional protected forms. Similarly, a reference to an amino group includes the protonated form (-N + HR 1 R 2 ), salts or solvates of amino groups (e.g., hydrochlorides), and conventional protected forms of amino groups. Similarly, references to hydroxy groups also include anionic forms (-O - ), salts or solvates thereof, as well as conventional protected forms.

[0491] Isomers, salts and solvates

[0492] Certain compounds may exist in one or more specific geometric, optical, enantiomeric, diastereomeric, epimeric, atropic, stereoisomeric, tautomeric, conformational or anomeric forms, including, but not limited to, cis- and trans-forms; E- and Z-forms; c-, t-, and r-forms; endo- and exo-forms; R-, S-, and meso-forms; D- and L-forms; d- and 1-forms; (+) and (-) forms; keto-, enol-, and enolate-forms; iso- and trans-forms; syncline- and anticline-forms; α- and β-forms; axial and equatorial forms; boat-, chair-, torsion-, envelope-, and hemi-chair-forms; and combinations thereof, hereinafter collectively referred to as "isomers" (or "isomeric forms").

[0493] Note that, in addition to the tautomeric forms discussed below, specifically excluded from the term "isomers" as used herein are structural (or constitutional) isomers (i.e., isomers that differ in the connectivity between the atoms rather than merely the position of the atoms in space). For example, a reference to a methoxy group -OCH3 is not to be construed as a reference to its structural isomer, the hydroxymethyl group -CH2OH.

[0494] Reference to a class of structures may also include structural isomeric forms belonging to that class (e.g., C 1-7 Alkyl includes n-propyl and isopropyl; butyl includes n-butyl, isobutyl, sec-butyl and tert-butyl; methoxyphenyl includes o-, m- and p-methoxyphenyl).

[0495] The above exclusion does not apply to tautomeric forms, such as keto-, enol- and enolate-forms, as in, for example, the following tautomeric pairs: keto / enol, imine / enamine, amide / iminoalcohol, amidine / amidine, nitroso / oxime, thione / enethiol, N-nitroso / hydroxyazo and nitro / isonitro.

[0496] Note that the term "isomer" specifically includes compounds having one or more isotopic substituents. For example, H can be in any isotopic form, including 1 H. 2 H(D) and 3 H(T); C can be in any isotopic form, including 12 C. 13 C and 14 C; O can be in any isotopic form, including 16 O and 18 O; etc.

[0497] Unless otherwise stated, a reference to a particular compound includes all such isomeric forms, including (wholly or partially) racemic and other mixtures thereof.

[0498] Methods for the preparation (eg, asymmetric synthesis) and separation (eg, fractional crystallization and chromatographic means) of such isomeric forms are known in the art or are readily obtained by modifying the methods taught herein or known methods in a known manner.

[0499] Unless otherwise indicated, a reference to a particular compound also includes its ionic, salt, solvate, and protected forms, such as those discussed below.

[0500] In some embodiments, the compounds of Formula (I) and salts and solvates thereof comprise pharmaceutically acceptable salts of the compounds of Formula (I).

[0501] The compounds of formula (I), including those specifically identified above, can form pharmaceutically acceptable complexes, salts, solvates, and hydrates. These salts include non-toxic acid addition salts (including diacids) and basic salts.

[0502] If the compound is cationic or has a functional group that can be cationic (e.g. -NH2 can be -NH3 + ), acid addition salts can be formed with suitable anions. Examples of suitable inorganic anions include, but are not limited to, those derived from the following inorganic acids: hydrochloric acid, nitric acid, nitrous acid, phosphoric acid, sulfuric acid, sulfurous acid, hydrobromic acid, hydroiodic acid, hydrofluoric acid, phosphoric acid, and phosphorous acid. Examples of suitable organic anions include, but are not limited to, those derived from the following organic acids: 2-acetoxybenzoic acid, acetic acid, ascorbic acid, aspartic acid, benzoic acid, camphorsulfonic acid, cinnamic acid, citric acid, edetic acid, ethanedisulfonic acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, hydroxymaleic acid, hydroxynaphthalenecarboxylic acid, hydroxyethanesulfonic acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, methanesulfonic acid, mucic acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, pantothenic acid, phenylacetic acid, benzenesulfonic acid, propionic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, sulfanilic acid, tartaric acid, toluenesulfonic acid, and valeric acid. Examples of suitable high molecular weight organic anions include, but are not limited to, those derived from the following high molecular weight acids: tannic acid, carboxymethyl cellulose. Such salts include acetate, adipate, aspartate, benzoate, benzenesulfonate, bicarbonate, carbonate, bisulfate, sulfate, borate, camphorsulfonate, citrate, cyclamate, edisylate, ethanesulfonate, formate, fumarate, glucoheptonic acid, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, methylsulfonate, naphthylate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate, hydrogenphosphate, dihydrogenphosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, and xinofoate.

[0503] For example, if the compound is anionic or has a functional group that can be anionic (e.g., -COOH can be -COO - ), then a basic salt can be formed with a suitable cation. Examples of suitable inorganic cations include, but are not limited to, metal cations (such as alkali metal or alkaline earth metal cations), ammonium and substituted ammonium cations, and amines. Examples of suitable metal cations include sodium (Na + ), potassium (K+ ), magnesium (Mg 2+ ), calcium (Ca 2+ ), zinc (Zn 2+ ) and aluminum (Al 3+ Examples of suitable organic cations include, but are not limited to, ammonium ions (ie, NH4 + ) and substituted ammonium ions (e.g., NH3R + NH2R2 + 、NHR3 + NR4 + Examples of some suitable substituted ammonium ions are those derived from ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine, and tromethamine, as well as amino acids such as lysine and arginine. An example of a commonly used quaternary ammonium ion is N(CH3)4 + Examples of suitable amines include arginine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethylamine, diethanolamine, dicyclohexylamine, ethylenediamine, glycine, lysine, N-methylglucamine, ethanolamine, 2-amino-2-hydroxymethyl-propane-1,3-diol, and procaine. For a discussion of useful acid addition salts or base salts, see SM Berge et al., J. Pharm. Sci. (1977) 66: 1-19; see also Stahl and Wermuth, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (2011).

[0504] Pharmaceutical salts can be prepared using various methods. For example, the compound of Formula 1 can be reacted with a suitable acid or base to obtain the desired salt. The precursor of the compound of Formula 1 can also be reacted with an acid or base to remove an acid-unstable or base-unstable protecting group or to open the lactone or lactam group of the precursor. In addition, the salt of the compound of Formula 1 can be converted into another salt by treating with a suitable acid or base or by contacting with an ion exchange resin. After the reaction, if the salt precipitates from the solution, the salt can be separated by filtration or recovered by evaporation. The degree of ionization of the salt can range from completely ionized to almost non-ionized.

[0505] It may be convenient or desirable to prepare, purify and / or process the corresponding solvates of the active compound. The term "solvate" describes a molecular complex comprising a compound and one or more pharmaceutically acceptable solvent molecules (e.g., EtOH). The term "hydrate" is a solvate in which the solvent is water. Pharmaceutically acceptable solvates include those in which the solvent may be isotopically substituted (e.g., D2O, acetone-d6, DMSO-d6).

[0506] For the solvates and hydrates of organic compounds, the currently accepted classification system is the classification system that distinguishes separation site, channel and metal-ion coordination solvates and hydrates.See, for example, KR Morris (HG Rittain ed.) Polymorphism in Pharmaceutical Solids (1995). Separation site solvates and hydrates are solvates and hydrates in which solvent (e.g., water) molecules are separated from each other by direct contact by the molecules of the organic compound being inserted. In channel solvates, solvent molecules are located in lattice channels where they are adjacent to other solvent molecules. In metal-ion coordination solvates, solvent molecules are bonded to metal ions.

[0507] When the solvent or water is tightly bound, the complex will have a well-defined stoichiometry, independent of humidity. However, when the solvent or water is weakly bound, as in channel solvates and hygroscopic compounds, the water or solvent content will depend on humidity and drying conditions. In this case, non-stoichiometry will generally be observed.

[0508] Compounds of Formula I include imine, carbinolamine, and carbinolamine ether forms of PDD. When a nucleophilic solvent (HO, ROH) is added across the imine bond of the PDD moiety, carbinolamine or carbinolamine ether is formed. The balance of these equilibria between these forms depends on the conditions in which the compound exists and the nature of the moiety itself.

[0509] These compounds can be isolated in solid form, for example by freeze-drying.

[0510] Further particular and preferred aspects are set out in the accompanying independent and dependent claims. Features of the dependent claims may be combined with features of the independent claims as appropriate and in combinations other than those explicitly set out in the claims.

[0511] Synthesis strategy

[0512] The compound of formula (I) can be prepared using the technology described below. Some schemes and examples can omit the details of common reactions, including oxidation, reduction, etc., separation techniques (extraction, evaporation, precipitation, chromatography, filtration, grinding, crystallization, etc.), and analytical procedures known to those of ordinary skill in the field of organic chemistry. The details of the reaction and technology can be found in many papers, including Richard Larock, Comprehensive Organic Transformations.A Guide to Functional Group Preparations, 2nd edition (2010), and the multi-volume series edited by Michael B.Smith and others, Compendium of Organic Synthetic Methods (1974 et seq.). Raw materials and reagents can be obtained from commercial sources or can be prepared using literature methods. Some reaction schemes can omit the minor products (for example, alcohol from ester hydrolysis, CO from diacid decarboxylation) produced by chemical transformations. In addition, in some cases, reaction intermediates can be used in subsequent steps (i.e., original position) without separation or purification.

[0513] In following some reaction schemes and example, some compound can use protecting group to prepare, and described protecting group prevents the undesirable chemical reaction at other reaction sites.Protecting group also can be used for enhancing solubility or otherwise changing the physical property of compound.For the discussion of protecting group strategy, for installing and removing the description of the materials and methods of protecting group, and the compilation of the available protecting group of common functional groups (comprising amine, carboxylic acid, alcohol, ketone, aldehyde etc.), referring to TWGreene and P.G.Wuts, Protecting Groups in Organic Chemistry, the 4th edition, (2006) and P.Kocienski, Protective Groups, the 3rd edition (2005).

[0514] In general, the chemical transformations described throughout the specification can be performed using substantially stoichiometric amounts of reactants, although certain reactions may benefit from the use of an excess of one or more reactants. Additionally, many reactions disclosed throughout the specification can be performed at about room temperature (RT) and ambient pressure, but some reactions may be performed under elevated pressure or using higher temperatures (e.g., reflux conditions) or lower temperatures (e.g., -78°C to 0°C), depending on reaction kinetics, yields, etc. Any reference to stoichiometric ranges, temperature ranges, pH ranges, etc. in the present disclosure, whether or not the word "range" is explicitly used, also includes the indicated endpoints.

[0515] Many chemical transformations can also utilize one or more compatible solvents that may affect the reaction rate and yield. Depending on the nature of the reactants, the one or more solvents can be polar protic solvents (including water), polar aprotic solvents, nonpolar solvents, or some combination. Representative solvents include saturated aliphatic hydrocarbons (e.g., n-pentane, n-hexane, n-heptane, n-octane); aromatic hydrocarbons (e.g., benzene, toluene, xylene); halogenated hydrocarbons (e.g., dichloromethane, chloroform, carbon tetrachloride); aliphatic alcohols (e.g., methanol, ethanol, propan-1-ol, propan-2-ol, butan-1-ol, 2-methyl-propan-1-ol, butan-2-ol, 2-methyl-propan-2-ol, pentan-1-ol, 3-methyl-butan-1-ol, hexan-1-ol, 2-methoxy-ethanol, 2- ethoxy-ethanol, 2-butoxy-ethanol, 2-(2-methoxy-ethoxy)-ethanol, 2-(2-ethoxy-ethoxy)-ethanol, 2-(2-butoxy-ethoxy)-ethanol); ethers (e.g., diethyl ether, diisopropyl ether, dibutyl ether, 1,2-dimethoxy-ethane, 1,2-diethoxy-ethane, 1-methoxy-2-(2-methoxy-ethoxy)-ethane, 1-ethoxy-2-(2-ethoxy-ethoxy)-ethane, tetrahydrofuran, 1,4 ... alkanes); ketones (e.g., acetone, methyl ethyl ketone); esters (methyl acetate, ethyl acetate); nitrogen-containing solvents (e.g., formamide, N,N-dimethylformamide, acetonitrile, N-methyl-pyrrolidone, pyridine, quinoline, nitrobenzene); sulfur-containing solvents (e.g., carbon disulfide, dimethyl sulfoxide, tetrahydro-thiophene-1,1,-dioxide); and phosphorus-containing solvents (e.g., hexamethylphosphoric triamide).

[0516] Further particular and preferred aspects are set out in the accompanying independent and dependent claims. Features of the dependent claims may be combined with features of the independent claims as appropriate and in combinations other than those explicitly set out in the claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0517] Embodiments of the present invention will now be further described with reference to the accompanying drawings, in which:

[0518] Figure 1 HPLC chromatograms are shown, providing evidence for the formation of DNA adducts with the NFκB transcription factor binding sequence using C8-linked PDD monomer 13;

[0519] Figure 2 HPLC chromatograms are shown, providing evidence for the formation of DNA adducts with the NFκB transcription factor binding sequence using C8-linked PDD monomer 17;

[0520] Figure 3HPLC chromatograms are shown, providing evidence for the formation of DNA adducts with the NFκB transcription factor binding sequence using C8-linked PDD monomer 20;

[0521] Figure 4 Shown is an HPLC chromatogram providing evidence for the formation of DNA adducts with the NFκB transcription factor binding sequence using the C8-linked PDD monomer 24. Example

[0522] General Notes

[0523] Synthetic building blocks and reagents were purchased from Maybridge Chemicals (UK), Fluorochem (USA), ChemShuttle Inc (USA), and Sigma-Aldrich (UK). Solvents were purchased from Sigma-Aldrich (UK) and Fisher Scientific (UK). Anhydrous reactions were performed in pre-dried glassware under an inert atmosphere of nitrogen. Anhydrous solvents were used as received without further drying. Thin layer chromatography (TLC) was performed on silica gel aluminum plates (Merck 60, F 254 ) and column chromatography was performed manually using silica gel (Merck 9385, 230-400 mesh ASTM, 40-63 μM) (while monitored by thin layer chromatography: UV (254 nm), and an alkaline aqueous solution of potassium permanganate as a stain) or using Grace The HPLC-MS / MS data were taken on a Bruker DPX400 spectrometer with a flow rate of 100 nm. The HPLC-MS / MS data were taken on a Waters Alliance 2695 HPLC coupled to a Waters 2695 HPLC with a flow rate of 0.1% formic acid and a column type of 50 × 4.60 mm. The HPLC-MS / MS data were taken on a Bruker DPX400 spectrometer with a flow rate of 0.1% formic acid and a column type of 50 × 4.60 mm. The HPLC-MS / MS data were taken on a Waters Alliance 2695 HPLC coupled to a Waters 2996 PDA. The Waters Micromass ZQ parameters used were: capillary (kV), 3.38; cone (V), 35; extractor (V), 3.0; source temperature (°C), 100; desolvation temperature (°C), 200; cone flow rate (L / h), 50; desolvation flow rate (L / h), 250. Microwave reactions were performed on an Anton Paar Monowave 300 microwave synthesis reactor. Unless otherwise stated, yields refer to isolated material (homogenized by TLC or NMR) and are named according to IUPAC nomenclature. LCMS gradient conditions are described below.

[0524] Method A (10 minutes): From 95% A / 5% B to 50% B in 3 minutes. Then from 50% B to 80% B in 2 minutes. Then from 80% B to 95% B in 1.5 minutes and hold constant for 1.5 minutes. Then reduce it to 5% B in 0.2 minutes and hold 5% B for 1.8 minutes. The flow rate is 0.5 mL / min and is divided into 200 μL via a zero dead volume T-piece and flows into the mass spectrometer. The wavelength range of the UV detector is 220-400 nm.

[0525] Method B (5 minutes): From 95% A / 5% B to 90% B in 3 minutes. Then from 90% B to 95% B in 0.5 minutes and hold constant for 1 minute. Then reduce it to 5% B in 0.5 minutes. The flow rate is 1.0 mL / min, divided into 100 μL portions via a zero dead volume T-piece and flowed into the mass spectrometer. The UV detector wavelength range is 220-500 nm.

[0526] General synthetic scheme

[0527]

[0528] i) K2CO3, DMF, methyl-4-bromobutyrate, rt; ii) KNO3, TFA, 0-5°C; iii) KMnO4, acetone, H2O, reflux; iv) oxalyl chloride, (S)-piperidin-2-ylmethanol, DMF cat, Et3N, CH2Cl2, 0°C-rt; v) H2, Pd / C, EtOH / EtOAc; vi) allyl chloroformate, pyridine, CH2Cl2, -10°C-rt; vii) TEMPO, BAIB, CH2Cl, rt; viiii) pTSA, DHP, EtOAc, rt; ix) NaOH, di Alkane, H2O, rt; x) RNH2, EDCl, DMAP, DMF, rt; xi) PPh3, Pd(PPh3)4, pyrrolidine, CH2Cl2, rt

[0529] Example 1: Methyl 4-(4-formyl-2-methoxyphenoxy)butyrate (1)

[0530]

[0531] A mixture of vanillin (20.0 g, 131 mmol), methyl 4-bromobutyrate (17.5 mL, 139 mmol) and potassium carbonate (27.2 g, 197 mmol) in N,N-dimethylformamide (100 mL) was stirred at room temperature for 18 hours. The reaction mixture was diluted with water (500 mL) and the title compound (30.2 g, 91%) was obtained by filtration as a white solid. The product was carried out to the next step without any further purification.

[0532] 1 H NMR (400MHz, CDCl3) δ9.84 (s, 1H), 7.46-7.37 (m, 2H), 6.98 (d, J=8.2Hz, 1H), 4.16 (t, J= 6.3Hz, 2H), 3.91 (s, 3H), 3.69 (s, 3H), 2.56 (t, J=7.2Hz, 2H), 2.20 (quin, J=6.7Hz, 2H); 13 C NMR (100MHz, CDCl3) δ190.9, 173.4, 153.8, 149.9, 130.1, 126.8, 111.6, 109.2, 67.8, 56.0, 51.7, 30.3, 24.2; MS m / z (EIMS) = 271.9 (M+Na) + ; LCMS (Method A): t R =6.48min.

[0533] Example 2: Methyl 4-(4-formyl-2-methoxy-5-nitrophenoxy)butyrate (2)

[0534]

[0535] To potassium nitrate (10.0g, 98.9mmol) in TFA (50mL) stirred solution was added dropwise 4- (4-formyl-2-methoxyphenoxy) butyric acid methyl ester (1) (20.0g, 79.2mmol) in TFA (50mL) at 0 ℃.Reaction mixture was stirred at room temperature for 1 hour.Then it was concentrated in a vacuum and diluted with ethyl acetate (400mL).Organic layer was washed with salt water (3x100mL) and a saturated aqueous solution of sodium bicarbonate (2x80mL), dried over sodium sulfate, filtered and concentrated to obtain title compound (23.5g, 100%), which is a yellow solid.Product was carried out to next step without any further purification.

[0536] 1 H NMR (400MHz, CDCl3) δ10.42 (s, 1H), 7.60 (s, 1H), 7.39 (s, 1H), 4.21 (t, J=6.3H z, 2H), 3.98 (s, 3H), 3.70 (s, 3H), 2.61-2.53 (m, 2H), 2.22 (quin, J=6.6Hz, 2H); 13 C NMR (100MHz, CDCl3) δ187.8, 173.2, 153.5, 151.7, 143.8, 125.5, 109.9, 108.1, 68.6, 56.6, 51.8, 30.2, 24.1; MS m / z (EIMS) = 296.1 (MH) - ; LCMS (Method A): t R =6.97min.

[0537] Example 3: 5-Methoxy-4-(4-methoxy-4-oxobutoxy)-2-nitrobenzoic acid (3)

[0538]

[0539] To a solution of 4-(4-formyl-2-methoxy-5-nitrophenoxy)butyric acid methyl ester (2) (23.0 g, 77.4 mmol) in acetone (600 mL) was quickly added a hot (70 ° C) solution of potassium permanganate (46.0 g, 291 mmol) in water (400 mL). The reaction mixture was stirred at 70 ° C for 3 hours. The reaction mixture was cooled to room temperature and passed through diatomaceous earth. The diatomaceous earth cake was washed with hot water (200 mL). A solution of sodium bisulfite in HCl (200 mL) was added to the filtrate and extracted with dichloromethane (2x400 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The obtained residue was purified by column chromatography (silica), eluted with methanol / dichloromethane (0% to 50%) to obtain the title compound (17.0 g, 70%) as a light yellow solid.

[0540] 1 H NMR (400MHz, MeOD) δ7.47 (s, 1H), 7.25 (s, 1H), 4.13 (t, J=6.2Hz, 2H), 3.94 (s, 3H), 3.68 (s, 3H), 2.54 (t, J=7.2Hz, 2H), 2.17-2.06 (m, 2H); 13 C NMR (100MHz, MeOD) δ175.3, 168.6, 153.8, 151.3, 143.1, 122.8, 112.4, 109.2, 69.6, 57.0, 52.2, 31.2, 25.5; MS m / z (EIMS) = 311.9 (MH) - ; LCMS (Method A): t R =6.22min.

[0541] Example 4: (S)-4-(4-(2-(hydroxymethyl)piperidine-1-carbonyl)-2-methoxy-5-nitrophenoxy)butanoic acid methyl Esters (4)

[0542]

[0543] A mixture of 5-methoxy-4-(4-methoxy-4-oxobutoxy)-2-nitrobenzoic acid (3) (8.0 g, 25.5 mmol), oxalyl chloride (6.6 mL, 77.0 mmol) and anhydrous N,N-dimethylformamide (2 drops) in anhydrous dichloromethane (100 mL) was stirred at room temperature for 1 hour. Anhydrous toluene (20 mL) was added to the reaction mixture, which was then concentrated in vacuo. A solution of the resulting residue in anhydrous dichloromethane (10 mL) was added dropwise to a solution of (S)-piperidin-2-ylmethanol (3.8 g, 33.4 mmol) and triethylamine (10.7 mL, 77.0 mmol) in anhydrous dichloromethane (90 mL) at -10°C. The reaction mixture was stirred at room temperature for 2 hours, then washed with hydrochloric acid (1 M, 50 mL) and brine (50 mL), dried over sodium sulfate, filtered and concentrated. The resulting residue was purified by column chromatography (silica) eluting with methanol / dichloromethane (0% to 5%) to give the title compound (9.2 g, 73%) as a yellow oil.

[0544] 1 H NMR (400MHz, CDCl3) δ7.68-7.64 (m, 1H), 6.77-6.70 (m, 1H), 4.16-4.07 (m, 3H), 3.93-3.89 (m, 3H), 3.83 (s, 1H), 3.6 7 (s, 3H), 3.15 (d, J = 1.4Hz, 1H), 3.11 (s, 1H), 2.78 (s, 1H), 2.56-2.50 (m, 3H), 2.21-2.12 (m, 4H), 1.74-1.55 (m, 4H); 13 C NMR (100MHz, CDCl3) δ173.3, 168.1, 154.6, 148.2, 137.4, 127.6, 111.4, 108.3, 68.3, 60.6, 56.7, 53.5, 51.7, 43.3, 38.0, 34.9, 30.3, 24.1, 19.7; MS m / z(EIMS)=411.0(M+H) + ; LCMS (Method A): t R =6.28min.

[0545] Example 5: (S)-4-(5-amino-4-(2-(hydroxymethyl)piperidine-1-carbonyl)-2-methoxyphenoxy)butanoic acid methyl Esters (5)

[0546]

[0547] To a solution of (S)-4-(4-(2-(hydroxymethyl)piperidine-1-carbonyl)-2-methoxy-5-nitrophenoxy)butanoic acid methyl ester (4) (9.2 g, 22.4 mmol) in ethanol (40 mL) and ethyl acetate (10 mL) was added palladium on activated carbon (10% basis weight) (920 mg). The reaction mixture was hydrogenated at 35 psi for 3 hours in a Parr apparatus. The reaction mixture was filtered through diatomaceous earth and the resulting cake was washed with ethyl acetate. The filtrate was concentrated in vacuo to give the title compound (9.0 g, 90%) as a pink solid. The product was carried on to the next step without any further purification.

[0548] 1 H NMR (400MHz, CDCl3) δ6.69 (s, 1H), 6.27-6.18 (m, 1H), 4.03-3.94 (m, 3H), 3.94-3.82 (m, 3H), 3.81-3.76 (m, 1H), 3.74 (s, 3H), 3.73-3.68 (m, 1H) ; 13 C NMR (100MHz, CDCl3) δ173.6, 171.2, 150.3, 141.8, 141.1, 113.2, 112.3, 102.4, 67.5, 60.8, 60.4, 56.8, 51.6, 30.4, 25.8, 24.3, 21.0, 19.9, 14.2; MSm / z (EIMS) = 381.0 (M+H) + ; LCMS (Method A): t R =5.52min.

[0549] Example 6: (S)-4-(5-(((allyloxy)carbonyl)amino)-4-(2-(hydroxy-methyl)piperidine-1-carbonyl)- Methyl 2-methoxyphenoxy)butyrate (6)

[0550]

[0551] To a solution of (S)-methyl 4-(5-amino-4-(2-(hydroxymethyl)piperidine-l- carbonyl)-2-methoxyphenoxy)butanoate (5) (9.0 g, 23.7 mmol) and pyridine (4.4 mL, 54.4 mmol) in anhydrous dichloromethane (100 mL) was added dropwise a solution of chloroformic acid allyl ester (2.6 mL, 24.8 mmol) in anhydrous dichloromethane (20 mL) at -10 °C. The reaction mixture was stirred at room temperature for 30 min. The reaction mixture was washed sequentially with saturated aqueous copper (II) sulfate (80 mL), water (80 mL) and saturated aqueous sodium bicarbonate (80 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The residue (2.0 g out of 11.0 g of crude product) was purified by column chromatography (silica) eluted with methanol / dichloromethane (0% to 1%) to give the title compound (930 mg, 47% based on the amount purified) as a yellow oil.

[0552] 1 H NMR (400 MHz, CDC13) δ 8.30 (br s, 1H), 7.63 (br s, 1H), 6.76 (br s, 1H), 5.92 (ddt, J = 17.2, 10.6, 5.4, 5.4 Hz, 1H), 5.37 - 5.28 (m, 1H), 5.20 (dq, J = 10.4, 1.3 Hz, 1H), 4.65 - 4.56 (m, 2H), 4.06 (t, J = 6.2 Hz, 2H), 3.94 - 3.82 (m, 1H), 3.79 (s, 3H), 3.66 (s, 3H), 3.62 - 3.54 (m, 1H), 3.40 (br s, 1H), 3.10 - 2.88 (m, 1H), 2.52 (t, J = 7.4 Hz, 2H), 2.22 - 2.04 (m, 3H), 1.64 (br s, 4H), 1.56 - 1.31 (m, 2H); 13 C NMR (100 MHz, CDC13) δ 173.5, 170.6, 153.9, 149.7, 144.8, 132.6, 130.1, 117.6, 116.9, 110.8, 107.1, 106.0, 67, 7, 65.6, 60.7, 56.3, 53.5, 51.6, 43.1, 30.5, 25.7, 24.4, 19.7; MS m / z (EIMS) = 465.1 (M+H) + LCMS (Method A): t R = 6.47 min.

[0553] Example 7: (6aS)-6-hydroxy-2-methoxy-3-(4-methoxy-4-oxobutoxy)-12-oxo-6,6a, 7,8,9,10-Hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -5(12H)-allyl formate (7)

[0554]

[0555] To a solution of (S)-4-(5-(((allyloxy)carbonyl)amino)-4-(2-(hydroxymethyl)-piperidine-1-carbonyl)-2-methoxyphenoxy)butanoic acid methyl ester (6) (930 mg, 2.0 mmol) in dichloromethane (45 mL) was added TEMPO (32 mg, 0.20 mmol) and (diacetoxyiodine)-benzene (773 mg, 2.4 mmol). The reaction mixture was stirred at room temperature for 16 hours and then washed sequentially with a saturated aqueous solution of sodium metabisulfite (20 mL), a saturated aqueous solution of sodium bicarbonate (20 mL), water (20 mL) and brine (20 mL). The organic layer was then dried over sodium sulfate, filtered and concentrated. The resulting residue was purified by column chromatography (silica) eluted with methanol / dichloromethane (0% to 5%) to obtain the title compound (825 mg, 89%) as a milky white solid.

[0556] MS m / z (EIMS) = 462.9 (M+H) + ; LCMS (Method A): t R =6.30min.

[0557] Example 8: (6aS)-2-methoxy-3-(4-methoxy-4-oxobutoxy)-12-oxo-6-((tetrahydro-2H- pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido-[1,2-a][1,4]diazepine -5(12H)- Allyl formate (8)

[0558]

[0559] (6aS)-6-hydroxy-2-methoxy-3-(4-methoxy-4-oxobutoxy)-12-oxo-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine A mixture of -5 (12H)-allyl formate (7) (825 mg, 1.8 mmol), 3,4-dihydro-2H-pyrans (1.7 mL, 18.2 mmol) and pTSA (8.5 mg, 1% w / w) in ethyl acetate (12 mL) was stirred at room temperature for 16 hours. The reaction mixture was then diluted with ethyl acetate (50 mL) and washed with a saturated aqueous solution of sodium bicarbonate (20 mL) and brine (30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The resulting residue was purified by column chromatography (silica), eluted with methanol / dichloromethane (0% to 2%) to obtain the title compound (820 mg, 84%) as a milky white solid.

[0560] MS m / z (EIMS) = 546.7 (M+H)+ ; LCMS (Method A): t R =7.70min.

[0561] Example 9: 4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyrrolidone)- pyrido[1,2-a][1,4]diazepine -3-yl) (9) butyric acid

[0562]

[0563] To (6aS)-2-methoxy-3-(4-methoxy-4-oxobutoxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -5(12H)-allylformate (8) (770 mg, 1.4 mmol) in 1,4-dimethoxybenzylformate To the 4-nitro-1-oxo-2-oxo-4 ...

[0564] MS m / z (EIMS) = 532.9 (M+H) + ; LCMS (Method A): t R =6.98min.

[0565] Example 10: 5-(4-((tert-Butyloxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)benzo[b] Thiophene-2-carboxylic acid methyl ester (10)

[0566]

[0567] By 4-((tert-butoxycarbonyl) amino)-1-methyl isophthalic acid H-pyrrole-2-formic acid (500mg, 2.1mmol) in N, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (725mg, 3.8mmol) and 4-(dimethylamino) pyridine (577mg, 4.7mmol) in the solution in dimethylformamide (10mL).Reaction mixture was stirred at room temperature for 2 hours.Then add 5-aminobenzo [b] thiophene-2-methylformate (392mg, 1.9mmol) and gained mixture was stirred at room temperature for 16 hours.Then pour it in frozen water (20mL) and extract with ethyl acetate (3x50mL).The organic extract merged is washed with 1M citric acid (30mL), sodium bicarbonate saturated aqueous solution (35mL), water (35mL) and salt solution (35mL) in sequence.Then organic layer is dried over sodium sulfate, filtered and concentrated. The resulting residue was purified by column chromatography (silica) eluting with ethyl acetate / hexanes (0% to 50%) to give the title compound (610 mg, 75%) as a beige solid.

[0568] MS m / z (EIMS) = 430.2 (M+H) + ; LCMS (Method A): t R =7.90min.

[0569] Example 11: 5-(4-amino-1-methyl-1H-pyrrole-2-carboxamido)-benzo[b]thiophene-2-carboxylic acid methyl ester Hydrochloride (11)

[0570]

[0571] Methyl 5-(4-((tert-butoxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)benzo-[b]thiophene-2-carboxylate (10) (610 mg, 1.4 mmol) was dissolved in hydrochloric acid (4 M in distilled water). The product was added to 4% paraformaldehyde (5% paraformaldehyde) (3.6 mL) and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated in vacuo to give the title compound (600 mg, 99%) as a brown solid. The product was carried on to the next step without any further purification.

[0572] MS m / z (EIMS) = 329.9 (M+H) + ; LCMS (Method A): t R =5.52min.

[0573] Example 12: (6aS)-2-methoxy-3-(4-((5-((2-(methoxycarbonyl)benzo-[b]thiophen-5-yl)amino (methyl-1H-pyrrol-3-yl)amino)-4-oxo-butoxy)-12-oxo-6-((tetrahydro-2H-pyran- 2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -5(12H)-formene Propyl ester (12)

[0574]

[0575] 4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To the product of 4- (4-amino-1-methyl-1H-pyrroles-2-formamido) benzo [b] thiophene-2-carboxylic acid methyl ester hydrochloride (11) (95 mg, 0.26 mmol) was added 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (100 mg, 0.52 mmol) and 4- (dimethylamino) pyridine (80 mg, 0.65 mmol) in a solution of N, N- dimethylformamide (4 mL). The reaction mixture was stirred at room temperature for 30 minutes. Then 5- (4-amino-1-methyl -1H-pyrroles-2-formamido) benzo [b] thiophene-2-carboxylic acid methyl ester hydrochloride (11) (95 mg, 0.26 mmol) was added, and the resulting mixture was stirred at room temperature for 16 hours. Then it was poured into frozen water (20 mL) and extracted with ethyl acetate (3x50 mL). The organic extracts merged were washed sequentially with 1M citric acid (30 mL), a saturated aqueous solution of sodium bicarbonate (35 mL), water (35 mL) and salt solution (35 mL). The organic layer was then dried over sodium sulfate, filtered and concentrated in vacuo to give the title compound (190 mg, 87%) as a yellow oil. The product was carried on to the next step without any further purification.

[0576] MS m / z (EIMS) = 844.0 (M+H) + ; LCMS (Method A): t R =8.10min.

[0577] Example 13: (S)-5-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrrolidone) pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-pyrrole-2-carboxamido)benzo [b]Methyl thiophene-2-carboxylate (13)

[0578]

[0579] To (6aS)-2-methoxy-3-(4-((5-((2-(methoxycarbonyl)benzo[b]-thiophen-5-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]-diazepine To a solution of -5 (12H)-allyl formate (12) (190 mg, 0.22 mmol) in dichloromethane (10 mL) was added tetrakis (triphenylphosphine) palladium (0) (13 mg, 5 mol%), triphenylphosphine (15 mg, 25 mol%) and pyrrolidine (22 μ L, 0.27 mmol). The reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was subjected to high vacuum for 30 minutes until excessive pyrrolidine was thoroughly removed. The resulting residue was then purified by column chromatography (silica) and eluted with acetone / dichloromethane (0% to 70%) to obtain the title compound (60 mg, 40%) as a yellow solid.

[0580] 1 H NMR (CDCl3, 400MHz) δ8.35 (s, 1H), 8.28 (s, 1H), 8.02 (s, 1H), 7.94 (s, 1H), 7.90 (d, J=5.7Hz, 1H), 7.75 (d , J=8.8Hz, 1H), 7.58 (dd, J=8.7, 2.1Hz, 1H), 7.42-7.41 (m, 1H), 7.13 (d, J=1.6Hz, 1H), 6.78 (s, 1H), 6.56 ( d, J=1.6Hz, 1H), 4.25-4.18 (m, 1H), 4.08 (t, J=6.0Hz, 2H), 3.93 (s, 3H), 3.88 (s, 3H), 3.83 (s, 3H), 3.79- 3.75 (m, 1H), 3.23-3.16 (m, 1H), 2.52-2.47 (m, 2H), 2.21 (d, J = 6.4Hz, 1H), 2.18 (d, J = 2.1Hz, 1H), 1.96 (br s, 2H), 1.86-1.81 (m, 2H), 1.77-1.66 (m, 2H); 13 C NMR (100MHz, CDCl3) δ170.0, 167.6, 163.4, 163.2, 160.0, 150.7, 148.0, 140.0, 139.2, 137.6, 135.8, 134.2, 130.6, 123.0, 122. 9,121.5,121.0,120.1,116.2,111.7,110.3,104.3,68.1,56.1,53.5,52.5,49.7,40.0,36.8,33.0,24.9,24.5,22.9,18.3;MS m / z(EIMS)=658.0(M+H) + ; LCMS (Method A): t R =6.92min.

[0581] Example 14: (6aS)-3-(4-((2-(ethoxycarbonyl)-1-methyl-1H-imidazol-4-yl)amino)-4-oxo butyloxy)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)-oxy)-6,6a,7,8,9,10-hexahydrobenzo [e]pyrido[1,2-a][1,4]diazepine -5(12H)-allyl formate (14)

[0582]

[0583] 4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To the product of 4- (4-amino-3-yl) oxygen base) butyric acid (9) (340mg, 0.64mmol) in N, 1- (3-dimethylaminopropyl) -3- ethylcarbodiimide hydrochloride (222mg, 1.2mmol) and 4- (dimethylamino) pyridine (177mg, 1.4mmol) solution in dimethylformamide (10mL). The reaction mixture was stirred at room temperature for 30 minutes. Then 4-amino-1-methyl -1H-imidazoles -2- ethyl formate hydrochloride (120mg, 0.58mmol) was added, and the gained mixture was stirred at room temperature for 16 hours. Then it was poured in frozen water (40mL) and extracted with ethyl acetate (3x100mL). The organic extract merged was washed sequentially with 1M citric acid (60mL), sodium bicarbonate saturated aqueous solution (70mL), water (70mL) and salt solution (70mL). The organic layer was then dried over sodium sulfate, filtered and concentrated in vacuo to give the title compound (350 mg, 80%) as a yellow oil. The product was carried on to the next step without any further purification.

[0584] MS m / z (EIMS) = 683.7 (M+H) + ; LCMS (Method A): t R =7.35min.

[0585] Example 15: 4-(4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro- 2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]-pyrido[1,2-a][1,4]diazepine - 3-amino)-1-methyl-1H-imidazole-2-carboxylic acid (15)

[0586]

[0587] To (6aS)-3-(4-((2-(ethoxycarbonyl)-1-methyl-1H-imidazol-4-yl)amino)-4-oxobutoxy)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -5(12H)-allylformate (14) (350 mg, 0.46 mmol) in 1,4-dimethoxybenzylformate To the mixture of 4-nitro-1-oxo-2-nitro-1-propane (10mL), 0.5M sodium hydroxide aqueous solution (10mL, 5.0mmol) was added to a solution in 4-nitro-1-oxo-2-nitro-1-propane (10mL). The reaction mixture was stirred at room temperature for 2 hours, then concentrated in a vacuum, then water (20mL) was added and the water layer was acidified to pH=1 with 1M citric acid solution (10mL). The water layer was then extracted with ethyl acetate (2x50mL). The organic extracts merged were then washed with salt water (50mL), dried over sodium sulfate, filtered and concentrated. The gained residue was ground in hexane, filtered and dried to obtain title compound (220mg, 74%), which was a beige solid. Product was carried out to the next step without any further purification.

[0588] MS m / z (EIMS) = 656.2 (M+H) + ; LCMS (Method A): t R =6.53min.

[0589] Example 16: (6aS)-2-methoxy-3-(4-((2-((2-(methoxycarbonyl)-benzo[b]thiophen-5-yl)amino (methylformyl)-1-methyl-1H-imidazol-4-yl)amino)-4-oxobutoxy)-12-oxo-6-((tetrahydro-2H-pyran-2- 1,2-a][1,4]diazepine -5(12H)-allylformate Esters (16)

[0590]

[0591] 4-(4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To the product was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (59 mg, 0.31 mmol) and 4-(dimethylamino)pyridine (47 mg, 0.38 mmol) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (59 mg, 0.31 mmol) in a solution in N,N-dimethylformamide (4 mL). The reaction mixture was stirred at room temperature for 30 minutes. Then 5-aminobenzo[b]thiophene-2-methylformate (32 mg, 0.15 mmol) was added and the resulting mixture was stirred at room temperature for 16 hours. The mixture was then poured into frozen water (40 mL) and extracted with ethyl acetate (3x100 mL). The organic extracts merged were washed sequentially with 1M citric acid (60 mL), a saturated aqueous solution of sodium bicarbonate (70 mL), water (70 mL) and salt solution (70 mL). The organic layer was then dried over sodium sulfate, filtered and concentrated. The resulting residue was then purified by column chromatography (silica), eluting with ethyl acetate / dichloromethane (0% to 100%) followed by methanol / dichloromethane (0% to 10%) to give the title compound (50 mg, 39%) as a yellow oil.

[0592] MS m / z (EIMS) = 844.9 (M+H) + ; LCMS (Method A): t R =8.22min.

[0593] Example 17: (S)-5-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrrolidone) pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-imidazole-2-carboxamido)benzo [b]Methyl thiophene-2-carboxylate (17)

[0594]

[0595]

[0596] To (6aS)-2-methoxy-3-(4-((2-((2-(methoxycarbonyl)benzo[b]-thiophen-5-yl)carbamoyl)-1-methyl-1H-imidazol-4-yl)amino)-4-oxobutoxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]-diazepine To a solution of -5 (12H)-allyl formate (16) (50 mg, 0.06 mmol) in dichloromethane (3 mL) was added tetrakis (triphenylphosphine) palladium (0) (3.5 mg, 5 mol%), triphenylphosphine (3.9 mg, 25 mol%) and pyrrolidine (5.8 μL, 0.07 mmol). The reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was subjected to high vacuum for 30 minutes until excess pyrrolidine was thoroughly removed. The resulting residue was then purified by column chromatography (silica) and eluted with acetone / dichloromethane (0% to 50%) to obtain the title compound (10 mg, 26%) as a yellow solid.

[0597] 1 H NMR (CDCl3, 400MHz) δ9.07 (s, 1H), 8.36 (d, J = 2.0Hz, 1H), 8.13 (s, 1H), 8.03 (s, 1H), 7.90 (d, J = 5.7Hz, 1H), 7 .82 (d, J=8.7Hz, 1H), 7.56 (dd, J=8.7, 2.1Hz, 1H), 7.49-7.43 (m, 2H), 6.81 (s, 1H), 4.26-4.17 (m, 2H), 4.10- 4.06 (m, 3H), 3.98-3.93 (m, 6H), 3.93-3.85 (m, 1H), 3.74 (td, J=5.8, 4.0Hz, 1H), 3.27-3.16 (m, 1H), 2.68-2. 60 (m, 2H), 2.29 (quin, J=6.4Hz, 2H), 2.10-2.02 (m, 1H), 1.97-1.89 (m, 1H), 1.83-1.77 (m, 2H), 1.76 (s, 2H); 13 C NMR (100MHz, CDCl3) δ169.7, 167.5, 163.3, 163.2, 160.3, 156.7, 150.4, 148.0, 140.0, 139.3, 135.8, 135.0, 130.6, 123.2 ,120.1,115.4,114.9,110.3,98.0,67.8,65.2,56.1,52.6,49.6,39.8,35.9,32.9,31.0,29.3,24.7,24.6,22.9,18.4; MS m / z(EIMS)=659.1(M+H) + ; LCMS (Method A): t R =7.00min.

[0598] Example 18: 4-(4-aminophenyl)-1-methyl-1H-pyrrole-2-carboxylic acid methyl ester (18)

[0599]

[0600] A mixture of 4-bromo-1-methyl-1H-pyrrole-2-carboxylic acid methyl ester (750 mg, 3.44 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)aniline (905 mg, 4.13 mmol) and potassium carbonate (1.43 g, 10.3 mmol) in toluene / ethanol / water (9:3:1) (13 mL in total) was degassed with nitrogen for 5 minutes. Tetrakis(triphenylphosphine)palladium(0) (230 mg, 6 mol%) was then added and the reaction mixture was irradiated with microwaves at 100°C for 15 minutes. Water (10 mL) was then added to the reaction mixture, which was extracted with ethyl acetate (3x40 mL). The combined organic extracts were then dried over sodium sulfate, filtered, and concentrated. The resulting residue was purified by column chromatography (silica) eluting with ethyl acetate / hexanes (0% to 50%) to give the title compound (145 mg, 18%) as a yellow solid.

[0601] MS m / z (EIMS) = 230.9 (M+H) + ; LCMS (Method A): t R =5.17min.

[0602] Example 19: (6S, 6aS)-2-methoxy-3-(4-((2-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrolidone) (1-Methyl-1H-imidazol-4-yl)amino)-4-oxobutyloxy)-12-oxo-6- ((Tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -5(12H)-Allyl formate (19)

[0603]

[0604] 4-(4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine 4-(4-aminophenyl)-1-methyl-1H-pyrrole-2-formic acid methyl ester (18) (35mg, 0.15mmol) and gained mixture were stirred at room temperature for 16 hours.Then it was poured in frozen water (40mL) and extracted with ethyl acetate (3x100mL).The organic extract merged was washed with 1M citric acid (60mL), sodium bicarbonate saturated aqueous solution (70mL), water (70mL) and salt solution (70mL) in sequence. The organic layer was then dried over sodium sulfate, filtered and concentrated.The resulting residue was then purified by column chromatography (silica) eluting with acetone / dichloromethane (0% to 50%) to give the title compound (54 mg, 37%) as a yellow oil.

[0605] MS m / z (EIMS) = 868.1 (M+H) + ; LCMS (Method A): t R =8.22min.

[0606] Example 20: (S)-4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e] Pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-imidazole-2-carboxamido)benzene 1-Methyl-1H-pyrrole-2-carboxylic acid methyl ester (20)

[0607]

[0608] To (6S,6aS)-2-methoxy-3-(4-((2-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-imidazol-4-yl)amino)-4-oxobutoxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of -5 (12H)-allyl formate (19) (54 mg, 0.06 mmol) in dichloromethane (3 mL) was added tetrakis (triphenylphosphine) palladium (0) (3.6 mg, 5 mol%), triphenylphosphine (4.1 mg, 25 mol%) and pyrrolidine (6.2 μL, 0.07 mmol). The reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was subjected to high vacuum for 30 minutes until excess pyrrolidine was thoroughly removed. The resulting residue was then purified by column chromatography (silica) and eluted with acetone / dichloromethane (0% to 50%) to obtain the title compound (22 mg, 52%) as a yellow solid.

[0609] 1 H NMR (CDCl3, 400MHz) δ8.95 (s, 1H), 8.27 (s, 1H), 7.89 (d, J=5.7Hz, 1H), 7.59 (d, J=8.6Hz, 2H), 7.47-7.41 (m, 4H) , 7.19 (d, J = 2.0Hz, 1H), 7.05 (d, J = 1.9Hz, 1H), 6.79 (s, 1H), 4.25-4.18 (m, 1H), 4.17-4.12 (m, 1H), 4.12-4.06 (m, 1H), 4.04(s, 3H), 3.95(s, 3H), 3.93(s, 3H), 3.84(s, 3H), 3.76-3.71(m, 1H), 3.26-3.16(m, 1H), 2.65-2.57(m, 2H ), 2.26 (t, J=6.4Hz, 2H), 2.09-2.01 (m, 2H), 1.96-1.89 (m, 1H), 1.85-1.77 (m, 2H), 1.67 (dd, J=10.9, 5.5Hz, 1H); 13 C NMR (100MHz, CDCl3) δ169.7, 167.5, 163.3, 161.7, 156.5, 150.4, 148.0, 140.0, 135.8, 135.6, 133.7, 130.6, 126.1, 125.5, 123 .1、122.8、120.0、114.6、111.6、110.2、67.8、56.1、51.2、49.6、39.8、37.0、35.8、32.8、31.0、29.7、24.7、24.5、22.9、18.4 m / z(EIMS)=682.1(M+H) + ; LCMS (Method A): t R =7.03min.

[0610] Example 21: (6aS)-2-methoxy-3-(4-((5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)amino 1-Hydroxy-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[3-((tetrahydro-2H-pyran-2-yl)oxy)-4-oxobutyloxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[3-( [e]pyrido[1,2-a][1,4]diazepine -5(12H)-allyl formate (21)

[0611]

[0612] 4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To the product of 4- (4-amino-3-yl) oxygen base) butyric acid (9) (150mg, 0.64mmol) in DMF (4mL) solution, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (98mg, 0.51mmol) and 4-(dimethylamino) pyridine (79mg, 0.64mmol). The reaction mixture was stirred at room temperature for 30 minutes. Then add 4-amino-1-methyl-1H-pyrroles-2-methyl-formiate hydrochloride (49mg, 0.26mmol), and the gained mixture was stirred at room temperature for 16 hours. Then it was poured in frozen water (40mL) and extracted with ethyl acetate (3x100mL). The organic extract merged was washed with 1M citric acid (60mL), sodium bicarbonate saturated aqueous solution (70mL), water (70mL) and salt solution (70mL) in sequence. The organic layer was then dried over sodium sulfate, filtered and concentrated in vacuo to give the title compound (150 mg, 88%) as a yellow oil. The product was carried on to the next step without any further purification.

[0613] MS m / z (EIMS) = 668.8 (M+H) + ; LCMS (Method A): t R =7.42min.

[0614] Example 22: 4-(4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro- 2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]-pyrido[1,2-a][1,4]diazepine - 3-amino)-1-methyl-1H-pyrrole-2-carboxylic acid (22)

[0615]

[0616] To (6aS)-2-methoxy-3-(4-((5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -5(12H)-allylformate (21) (150 mg, 0.22 mmol) in 1,4-dimethoxybenzylamine To the 4-nitro-1-oxo-2-oxo-4 ...

[0617] MS m / z (EIMS) = 677.0 (M+Na) + ; LCMS (Method A): t R =6.92min.

[0618] Example 23: (6S, 6aS)-2-methoxy-3-(4-((5-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrolidone) (1-Methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyloxy)-12-oxo-6- ((Tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -5(12H)-allyl formate (23)

[0619]

[0620] 4-(4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To the product was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (74mg, 0.39mmol) and 4-(dimethylamino)pyridine (59mg, 0.48mmol) of 4-(4-aminophenyl)-1-methyl-1H-pyrrole-2-formic acid methyl ester (18) (45mg, 0.19mmol) at room temperature. The obtained mixture was stirred for 16 hours. The mixture was poured into frozen water (40mL) and extracted with ethyl acetate (3x100mL). The organic extracts merged were washed with 1M citric acid (60mL), a saturated aqueous solution of sodium bicarbonate (70mL), water (70mL) and salt solution (70mL) in sequence. The organic layer was then dried over sodium sulfate, filtered and concentrated.The resulting residue was then purified by column chromatography (silica) eluting with acetone / dichloromethane (0% to 50%) to give the title compound (160 mg, 95%) as a yellow solid.

[0621] MS m / z (EIMS) = 867.0 (M+H) + ; LCMS (Method A): t R =8.10min.

[0622] Example 24: (S)-4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e] Pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-pyrrole-2-carboxamido)benzene 1-Methyl-1H-pyrrole-2-carboxylic acid methyl ester (24)

[0623]

[0624]

[0625] To (6S,6aS)-2-methoxy-3-(4-((5-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of -5(12H)-allyl formate (23) (80 mg, 0.09 mmol) in dichloromethane (3 mL) was added tetrakis(triphenylphosphine)palladium(0) (5.3 mg, 5 mol%), triphenylphosphine (6.1 mg, 25 mol%) and pyrrolidine (9.1 μL, 0.11 mmol). The reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was subjected to high vacuum for 30 minutes until excess pyrrolidine was thoroughly removed. The resulting residue was then purified by column chromatography (silica) eluted with acetone / dichloromethane (0% to 50%) to obtain the title compound (23 mg, 37%) as a yellow solid.

[0626] 1H NMR (CDCl3, 400MHz) δ8.09 (s, 1H), 8.04-8.01 (m, 1H), 7.90 (d, J=5.8Hz, 1H), 7.58 (s, 1H), 7.56 (s, 1H), 7.44-7.40 (m, 3H) , 7.18 (d, J = 2.0Hz, 1H), 7.12 (d, J = 1.8Hz, 1H), 7.04 (d, J = 2.0Hz, 1H), 6.78 (s, 1H), 6.50 (d, J = 1.9Hz, 1H), 4.26-4.18 (m, 1 H), 4.07 (t, J=6.0Hz, 2H), 3.94 (s, 3H), 3.87 (s, 3H), 3.84 (d, J=2.9Hz, 6H), 3.76 (td, J=5.7, 3.9Hz, 1H), 3.25-3.15 (m, 1H ), 2.49(t, J=7.0Hz, 2H), 2.24-2.18(m, 2H), 2.10-2.03(m, 1H), 2.01-1.93(m, 2H), 1.86-1.80(m, 2H), 1.73-1.66(m, 1H); 13 C NMR (100MHz, CDCl3) δ169.9, 167.6, 163.5, 161.7, 159.7, 150.7, 147.9, 139.9, 136.4, 130.2, 126.1, 125.4, 123.3, 123.0, 120.6 MS m / z(EIMS)=681.0(M+H) + ; LCMS (Method A): t R =6.98min.

[0627] Example 25: (6aS)-2-methoxy-3-(4-((4-(methoxycarbonyl)phenyl)-amino)-4-oxobutoxy yl)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2- a][1,4]diazepine -5(12H)-allyl formate (25)

[0628]

[0629]

[0630] 4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of 1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanamine was added 1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanamine. To the 4-aminobenzoic acid saturated aqueous solution (20mL) of 4-aminobenzoic acid (57.0mg, 0.376mmol) of 4-aminobenzoic acid (150mg, 0.394mmol) and anhydrous triethylamine (220 μ L, 1.58mmol).Reaction mixture was at room temperature stirred for 30 minutes.Then 4-aminobenzoic acid methyl ester (57.0mg, 0.376mmol) was added and the gained mixture was at room temperature stirred for 16 hours.Reaction mixture was quenched with sodium bicarbonate saturated aqueous solution (20mL) and extracted with dichloromethane (2x50mL).The organic extract merged was washed with the water (30mL) containing a few drops of acetic acid.Then the organic layer was dried over sodium sulfate, filtered and concentrated in a vacuum.Then by column chromatography (silica) purification of the obtained residue, eluted with methanol / dichloromethane (0% to 10%), to obtain title compound (110mg, 44%), it is a yellow solid.

[0631] MS (ES+): m / z = 666 (M+H) + ; LCMS (Method A): t R =7.88min.

[0632] Example 26: 4-(4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro- 2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]-pyrido[1,2-a][1,4]diazepine - 3-(3-amino)-1-methyl)-1-butyl-1-amino)-1-butyl-1-amino)-benzoic acid (26)

[0633]

[0634] To (6aS)-2-methoxy-3-(4-((4-(methoxycarbonyl)phenyl)amino)-4-oxobutoxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo-[e]pyrido[1,2-a][1,4]diazepine -5(12H)-allylformate (25) (90 mg, 0.14 mmol) in 1,4-dimethoxybenzylformate To the 4-nitro-1-oxo-2-oxo-4 ...

[0635] MS (ES+): m / z = 652 (M+H) + ; LCMS (Method A): t R =7.13min.

[0636] Example 27: (6aS)-2-methoxy-3-(4-((4-(4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrole- (3-yl)phenyl)carbamoyl)phenyl)amino)-4-oxobutyloxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl) Oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]-pyrido[1,2-a][1,4]diazepine -5(12H)-allyl formate (27)

[0637]

[0638] 4-(4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of (40 mg, 0.061 mmol) of 1H-1,2,3-triazolo-[4,5-b]-pyridin-1-ylmethylene)-N-methylmethanamine was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]-pyridin-1-ylmethylene]-N-methylmethanamine To the 4- (4-aminophenyl) -1- methyl -1H- pyrrole -2- carboxylic acid methyl ester (18) (14mg, 0.061mmol) of hexafluorophosphate N- oxide (25mg, 0.064mmol) and anhydrous triethylamine (36 μ L, 0.26mmol). The reaction mixture was stirred at room temperature for 30 minutes. Then 4- (4-aminophenyl) -1- methyl -1H- pyrrole -2- carboxylic acid methyl ester (18) (14mg, 0.061mmol) was added and the resulting mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with a saturated aqueous solution of sodium bicarbonate (20mL) and extracted with dichloromethane (2x50mL). The organic extracts merged were washed with water (30mL) containing a few drops of acetic acid. The organic layer was then dried over sodium sulfate, filtered and concentrated in a vacuum. The resulting residue was then purified by column chromatography (silica), eluted with methanol / dichloromethane (0% to 10%), to obtain title compound (43mg, 63%), which was a yellow oil.

[0639] MS (ES+): m / z = 864 (M+H) + ; LCMS (Method A): t R =8.10min.

[0640] Example 28: (S)-4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e] Pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-benzamido)phenyl)-1-methyl-1H-pyrrolidone Methyl pyrrole-2-carboxylate (28)

[0641]

[0642] To (6aS)-2-methoxy-3-(4-((4-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)phenyl)amino)-4-oxobutoxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of -5(12H)-allyl formate (27) (33 mg, 0.038 mmol) in dichloromethane (3 mL) was added tetrakis-(triphenylphosphine)palladium (0) (2.2 mg, 5 mol%), triphenylphosphine (2.5 mg, 25 mol%) and pyrrolidine (4 μL, 0.11 mmol). The reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was subjected to high vacuum for 30 minutes until excess pyrrolidine was thoroughly removed. The resulting residue was then purified by column chromatography (silica) eluted with acetone / dichloromethane (0% to 100%) to obtain the title compound (5.8 mg, 21%) as a yellow solid.

[0643] 1H NMR (CDCl3, 400MHz) δ8.14 (br s, 1H), 8.06 (br s, 1H), 7.91 (d, J=5.7Hz, 1H), 7.81 (d, J=8.7Hz, 2H), 7.65 (d, J=8.6Hz, 2H), 7.60 (d, J=8.4Hz, 2H), 7 .46-7.50 (m, 2H), 7.41 (s, 1H), 7.21 (d, J=2.1Hz, 1H), 7.08 (d, J=1.9Hz, 1H), 6.78-6.82 (m, 1H), 4.2 4(d, J=14.0Hz, 1H), 4.11-4.18(m, 2H), 3.95-3.98(m, 3H), 3.83-3.86(m, 6H), 3.74-3.79(m, 2H), 3. 18-3.30 (m, 2H), 2.60-2.66 (m, 2H), 2.28 (t, J=6.3Hz, 2H), 1.97 (d, J=6.3Hz, 2H), 1.82-1.88 (m, 2H); 13 C NMR (100MHz, CDCl3) δ215.5, 171.1, 167.5, 165.0, 163.4, 161.7, 150.3, 147.8, 141.3, 140.0, 136.2, 130.8, 128.1, 125.6, 123.5, 123.1, 1 21.5, 120.6, 119.3, 114.7, 111.7, 110.2, 67.9, 56.1, 51.2, 49.7, 39.8, 37.0, 34.3, 30.9, 25.6, 24.5, 23.0, 18.4; MS (ES+): m / z=678 (M+H) + ; LCMS (Method A): t R =7.05min.

[0644] Example 29: (6aS)-2-methoxy-3-(4-((4-((2-(methoxycarbonyl)benzo-[b]thiophen-5-yl)amino ((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7, 8,9,10-Hexahydrobenzo[e]pyrido[1,2-a]-[1,4]diazepine -5(12H)-Allyl formate (29)

[0645]

[0646] 4-(4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of (40 mg, 0.061 mmol) of 1H-1,2,3-triazolo-[4,5-b]-pyridin-1-ylmethylene)-N-methylmethanamine was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]-pyridin-1-ylmethylene]-N-methylmethanamine To the 4-thiazolinone (5-nitro-2-propene) of 4-nitro-1-propene) is dissolved in 4% paraffin wax and 5-nitro-2-propene) is added hexafluorophosphate N-oxide (25mg, 0.064mmol) and anhydrous triethylamine (36 μ L, 0.26mmol).Reaction mixture was at room temperature stirred for 30 minutes.Then 5-aminobenzo [b]-thiophene-2-methyl-formiate (13mg, 0.063mmol) was added and the gained mixture was at room temperature stirred for 16 hours.Reaction mixture was quenched with a saturated aqueous solution of sodium bicarbonate (20mL) and extracted with dichloromethane (2x50mL).The organic extract merged was washed with the water (30mL) containing a few drops of acetic acid.Then the organic layer was dried over sodium sulfate, filtered and concentrated in a vacuum.Then by column chromatography (silica) purification of the obtained residue, eluted with methanol / dichloromethane (0% to 10%), to obtain title compound (34mg, 45%), it is a brown oil.

[0647] MS (ES+): m / z = 841 (M+H) + ; LCMS (Method A): t R =8.15min.

[0648] Example 30: (S)-5-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrrolidone pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-benzamido)benzo[b]thiophene-2-carboxylic acid methyl Esters (30)

[0649]

[0650] To (6aS)-2-methoxy-3-(4-((4-((2-(methoxycarbonyl)benzo[b]-thiophen-5-yl)carbamoyl)phenyl)amino)-4-oxobutoxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of allyl-5(12H)-formate (29) (23 mg, 0.028 mmol) in dichloromethane (1.5 mL) was added tetrakis(triphenylphosphine)palladium(0) (1.6 mg, 5 mol%), triphenylphosphine (1.8 mg, 25 mol%) and pyrrolidine (3.0 μL, 0.11 mmol). The reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was subjected to high vacuum for 30 minutes until the excess pyrrolidine was completely removed. The resulting residue was then purified by column chromatography (silica) eluting with acetone / dichloromethane (0% to 100%) followed by methanol / dichloromethane (0% to 100%) to afford the title compound (5.4 mg, 30%) as a pink solid.

[0651] 1 H NMR (CDCl3, 400MHz) δ8.48 (br s, 1H), 8.39 (d, J=1.9Hz, 1H), 8.31 (s, 1H), 7.97 (s, 1H), 7.91 (d, J=5.8Hz, 1H), 7.77-7.84 (m, 3H), 7.65 (dd, J=8.8, 2.0Hz, 1H), 7.57 (d, J=8.6Hz, 2H), 7.38 (s, 1H), 6.79 (s, 1H), 4.24 (dt, J=13.7, 4.1Hz, 1H), 4.09-4.17 (m, 2H), 3.95 (s, 3H), 3.79-3.82 (m, 3H), 3.74-3.79 (m, 1H), 3.49 (d, J=3.9Hz, 1H), 3.29-3.41 (m, 1H), 3.17-3.28 (m, 1H), 2.58-2 .64 (m, 2H), 2.26 (quin, J=6, 2Hz, 2H), 2.05-2.13 (m, 1H), 1.92-2.01 (m, 1H), 1.83-1.87 (m, 1H), 1.07-1.19 (m, 1H); 13 C NMR (CDCl3, 100MHz) δ171.2, 167.5, 165.5, 163.4, 163.2, 150.4, 147.8, 141.5, 140.0, 139.3, 138.0, 135.1, 134.4, 130.6, 128.2, 123. 0, 121.4, 120.9, 119.2, 116.4, 111.7, 110.1, 67.9, 56.0, 52.6, 49.7, 39.8, 34.2, 30.9, 24.7, 24.5, 22.9, 18.3; MS (ES+): m / z=655 (M+H) + ; LCMS (Method A): t R =7.00min.

[0652] Example 31: Methyl 4-(4-((tert-Butoxycarbonyl)amino)-1-methyl-1H-imidazole-2-carboxamido)benzoate Esters (31)

[0653]

[0654] To a solution of 4-((tert-butoxycarbonyl)amino)-1-methyl-1H-imidazole-2-carboxylic acid (100 mg, 0.415 mmol) in anhydrous dichloromethane (3 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methyl-methylamine To the 4-aminobenzoic acid saturated aqueous solution (20mL) of 4-aminobenzoic acid (165mg, 0.435mmol) and anhydrous triethylamine (242 μ L, 1.74mmol).Reaction mixture was at room temperature stirred for 30 minutes.Then 4-aminobenzoic acid methyl ester (63mg, 0.42mmol) was added and the gained mixture was at room temperature stirred for 16 hours.Reaction mixture was quenched with sodium bicarbonate saturated aqueous solution (20mL) and extracted with dichloromethane (2x50mL).The organic extract merged was washed with the water (30mL) containing a few drops of acetic acid.Then the organic layer was dried over sodium sulfate, filtered and concentrated in a vacuum.Then by column chromatography (silica) purification of the obtained residue, eluted with methanol / dichloromethane (0% to 10%), to obtain title compound (40mg, 26%), it is a custard solid.

[0655] 1 H NMR (CDCl3, 400MHz) δ9.16 (s, 1H), 8.01-8.07 (m, 2H), 7.69-7.75 (m, 2H), 7.21 (br s, 1H), 6.84 (s, 1H), 4.07 (s, 3H), 3.92 (s, 3H), 1.53 (s, 9H); MS (ES-): m / z=373 (MH) - ; LCMS (Method A): t R =7.68min.

[0656] Example 32: 4-(4-Amino-1-methyl-1H-imidazole-2-carboxamido)-benzoic acid methyl ester hydrochloride (32)

[0657]

[0658] Methyl 4-(4-((tert-Butoxycarbonyl)amino)-1-methyl-1H-imidazole-2-carboxamido)-benzoate (31) (40 mg, 0.11 mmol) was dissolved in hydrochloric acid (4 M in 1,4-dihydrochloric acid). The mixture was added with 4% paraformaldehyde (2 mL) and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated in vacuo to give the title compound (33 mg, 99%) as a brown solid. The product was carried on to the next step without any further purification.

[0659] 1 H NMR (MeOD, 400MHz) δ7.89-7.95 (m, 2H), 7.72-7.78 (m, 2H), 7.31 (s, 1H), 4.01 (s, 3H), 3.80 (s, 3H); 13 C NMR (MeOD, 100MHz) δ168.0, 143.6, 132.5, 131.6, 126.9, 123.3, 120.6, 92.6, 68.1, 52.3, 36.7; MS (ES+): m / z=275 (M+H) + ; LCMS (Method A): t R =5.43min.

[0660] Example 33: (6aS)-2-methoxy-3-(4-((2-((4-(methoxycarbonyl)phenyl)-carbamoyl)-1- methyl-1H-imidazol-4-yl)amino)-4-oxobutyloxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6, 6a,7,8,9,10-Hexahydrobenzo[e]pyrido-[1,2-a][1,4]diazepine -5(12H)-allyl formate (33)

[0661]

[0662] 4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of 1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanamine was added 1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanamine Hexafluorophosphate N-oxide (38 mg, 0.099 mmol) and anhydrous triethylamine (55 μL, 0.40 mmol). The reaction mixture was stirred at room temperature for 30 minutes. Then methyl 4-(4-amino-1-methyl-1H-imidazole-2-carboxamido)- benzoate hydrochloride (32) (30 mg, 0.094 mmol) was added and the resulting mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with saturated aqueous sodium bicarbonate solution (20 mL) and extracted with dichloromethane (2 x 50 mL). The combined organic extracts were washed with water containing a few drops of acetic acid (30 mL). The organic layer was then dried over sodium sulfate, filtered and concentrated in vacuo. The resulting residue was then purified by column chromatography (silica) eluted with methanol / dichloromethane (0% to 10%) to give the title compound (72 mg, 97%) as a brown oil.

[0663] MS (ES+): m / z = 789 (M+H) + ; LCMS (Method A): t R = 7.87 min.

[0664] Example 34: (S)-4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrrolidone pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-imidazole-2-carboxamido)benzoic acid Methyl ester (34)

[0665]

[0666]

[0667] To a solution of (6aS)-2-methoxy-3-(4-((2-((4-(methoxycarbonyl)phenyl)- carbamoyl)-1-methyl-1H-imidazol-4-yl)amino)-4-oxobutoxy)-12-oxo-6-((tetrahydro- 2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5(12H)- carboxylic acid allyl ester (33) (72 mg, 0.091 mmol) in dichloromethane (2 mL) was added tetrakis(triphenylphosphine)palladium(0) (5.3 mg, 5 mol%), triphenylphosphine (6.0 mg, 25 mol%) and pyrrolidine (9.0 μL, 0.11 mmol). The reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was subjected to high vacuum for 30 minutes until excess pyrrolidine was completely removed. The resulting residue was then purified by column chromatography (silica) eluted with acetone / dichloromethane (0% to 100%) to give the title compound (15 mg, 27%) as a yellow solid.

[0668] ​1 H NMR(CDCl3,400MHz)δ9.13(s,1H),8.24(s,1H),8.03(d,J=8.7Hz,2H),7.90(d,J=5.7Hz,1H),7.65-7.75(m,2H),7.43-7.50(m,2H),6.77-6.83(m,1H),4.12-4.23(m,2H),4.07(s,3H),3.93(s,3H),3.91(s,3H),3.18-3.27(m,1H),2.80(s,3H),2.56-2.68(m,3H),2.23-2.31(m,2H),1.85(d,J=10.1Hz,4H); 13 C NMR(CDCl3,100MHz)δ169.3,167.1,166.2,162.9,156.2,150.1,147.6,147.4,141.4,139.6,135.6,132.8,130.5,130.3,125.2,121.1,118.2,114.8,111.2,109.9,94.1,67.4,63.5,55.7,53.4,51.6,49.2,39.4,38.2,35.5,32.5,31.6,30.9,28.9,24.9,24.3,24.1,22.5,19.9,18.0;MS(ES+):m / z=603(M+H) + ;LCMS(方法A):t R =6.57min。

[0669] Example 35: Methyl 4-(4-((tert-Butoxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)benzoate Esters (35)

[0670]

[0671] By 4-((tert-butoxycarbonyl) amino)-1-methyl isophthalic acid H-pyrrole-2-formic acid (100mg, 0.416mmol) in the solution in DMF (3mL), add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (145mg, 0.756mmol) and 4-(dimethylamino) pyridine (115mg, 0.941mmol).Reaction mixture was at room temperature stirred 3 hours.Then add 4-aminobenzoic acid methyl ester (57mg, 0.38mmol) and gained mixture was at room temperature stirred 16 hours.Then pour it in frozen water (40mL) and extract with ethyl acetate (3x100mL).The organic extract merged is washed with citric acid aqueous solution (1M, 60mL), sodium bicarbonate saturated aqueous solution (70mL), water (70mL) and salt solution (70mL) sequentially.Then organic layer is dried over sodium sulfate, filtered and concentrated. The resulting residue was then purified by column chromatography (silica) eluting with methanol / dichloromethane (0% to 10%) to give the title compound (90 mg, 58%) as a white solid.

[0672] 1 H NMR (CDCl3, 400MHz) δ7.99-8.07 (m, 2H), 7.69 (s, 1H), 7.61-7.67 (m, 2H), 6.88 (s, 1H), 6.69 (br s, 1H), 6.25 (br s, 1H), 3.93 (s, 3H), 3.91 (s, 3H), 1.52 (s, 9H); 13 C NMR (CDCl3, 100MHz) δ166.6, 159.4, 153.4, 142.3, 130.9, 125.5, 123.1, 122.5, 119.2, 118.7, 140.1, 80.5, 52.0, 36.8, 28.4; MS (ES+): m / z=374 (M+H) + ; LCMS (Method A): t R =7.52min.

[0673] Example 36: 4-(4-Amino-1-methyl-1H-pyrrole-2-carboxamido)-benzoic acid methyl ester hydrochloride (36)

[0674]

[0675] Methyl 4-(4-((tert-Butoxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)-benzoate (35) (90 mg, 0.24 mmol) was dissolved in hydrochloric acid (4 M in 1,4-dihydro- The product was added to 4% paraformaldehyde (5% paraformaldehyde) (3 mL) and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated in vacuo to give the title compound (79 mg, 99%) as a creamy white solid. The product was carried on to the next step without any further purification.

[0676] 1 H NMR (MeOD, 400MHz) δ7.99 (d, J=8.7Hz, 2H), 7.80 (d, J=8.7Hz, 2H), 7.13 (d, J=1.9Hz, 1H), 7.09 (d, J=1.9Hz, 1H), 3.96 (s, 3H), 3.89 (s, 3H); 13 C NMR (MeOD, 100MHz) δ168.2, 161.2, 144.5, 131.5, 126.9, 126.4, 123.7, 120.8, 114.2, 109.0, 52.5, 37.5; MS (ES+): m / z=274 (M+H) + ; LCMS (Method A): t R =4.98min.

[0677] Example 37: (6aS)-2-methoxy-3-(4-((5-((4-(methoxycarbonyl)phenyl)-carbamoyl)-1- methyl-1H-pyrrol-3-yl)amino)-4-oxobutyloxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6, 6a,7,8,9,10-Hexahydrobenzo[e]pyrido-[1,2-a][1,4]diazepine -5(12H)-Allyl formate (37)

[0678]

[0679] 4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of 1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanamine was added 1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanamine. To the 4- (4-amino-1-methyl-1H-pyrrole-2-formamido) methyl benzoate hydrochloride (36) (30 mg, 0.094 mmol) of hexafluorophosphate N-oxide (38 mg, 0.099 mmol) and anhydrous triethylamine (55 μ L, 0.40 mmol). The reaction mixture was stirred at room temperature for 30 minutes. Then 4- (4-amino-1-methyl-1H-pyrrole-2-formamido) methyl benzoate hydrochloride (36) (30 mg, 0.094 mmol) was added, and the resulting mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with a saturated aqueous solution of sodium bicarbonate (20 mL) and extracted with dichloromethane (2x50 mL). The organic extracts merged were washed with water (30 mL) containing a few drops of acetic acid. The organic layer was then dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was then purified by column chromatography (silica), eluted with methanol / dichloromethane (0% to 10%), to obtain title compound (72 mg, 97%), which was a brown oil.

[0680] MS (ES+): m / z = 788 (M+H) + ; LCMS (Method A): t R =7.77min.

[0681] Example 38: (S)-4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrrolidone pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-pyrrole-2-carboxamido)benzoic acid Methyl ester (38)

[0682]

[0683] To (6aS)-2-methoxy-3-(4-((5-((4-(methoxycarbonyl)phenyl)-carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of -5(12H)-allyl formate (37) (72 mg, 0.091 mmol) in dichloromethane (2 mL) was added tetrakis-(triphenylphosphine)palladium (0) (5.3 mg, 5 mol%), triphenylphosphine (6.0 mg, 25 mol%) and pyrrolidine (9.0 μL, 0.11 mmol). The reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was subjected to high vacuum for 30 minutes until excess pyrrolidine was thoroughly removed. The resulting residue was then purified by column chromatography (silica) and eluted with acetone / dichloromethane (0% to 100%) to obtain the title compound (15.0 mg, 27%) as a yellow solid.

[0684] 1H NMR (CDCl3, 400MHz) δ8.41 (s, 1H), 8.00 (s, 2H), 7.98 (s, 1H), 7.90 (d, J=5.8Hz, 1H), 7.72-7. 74 (m, 1H), 7.70-7.72 (m, 1H), 7.41 (s, 1H), 7.14 (d, J=1.8Hz, 1H), 6.79 (s, 1H), 6.57 (d, J=1. 8Hz, 1H), 4.22 (d, J=14.1Hz, 1H), 4.09 (t, J=6.0Hz, 2H), 3.89 (s, 3H), 3.88 (s, 3H), 3.83 (s, 3 H), 3.74-3.79 (m, 2H), 3.21 (d, J=3.3Hz, 1H), 2.47-2.52 (m, 2H), 2.17-2.23 (m, 2H), 1.93 (br s, 3H), 1.79-1.85 (m, 2H); 13 C NMR (CDCl3, 100MHz) δ170.0, 167.6, 166.8, 163.6, 159.8, 150.7, 147.9, 142.9, 139.9, 130.7, 124.9, 122.8, 121.6, 121.5, 120. 8, 119.1, 111.8, 110.4, 104.6, 68.1, 56.1, 52.0, 49.7, 39.9, 36.9, 33.0, 31.0, 25.0, 24.5, 22.9, 18.3; MS (ES+): m / z=602 (M+H) + ; LCMS (Method A): t R =6.52min.

[0685] Example 39: 4-(4-(4-(4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6- ((Tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]-pyrido[1,2-a][1,4]di Aza -3-yl)oxy)butyramido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2- Formic acid (39)

[0686]

[0687] To (6aS)-2-methoxy-3-(4-((5-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -5(12H)-allylformate (23) (195 mg, 0.225 mmol) in 1,4-dimethoxybenzylamine To the 4-nitro-1-oxo-2-oxo-4-oxo-2 ...

[0688] MS (ES+): m / z = 853 (M+H) + ; LCMS (Method B): t R =3.83min.

[0689] Example 40: (6aS)-3-(4-((5-((4-(5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrolidone (1-Methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-2-methoxy-12- Oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4] Diazepines -5(12H)-Allyl formate (40)

[0690]

[0691] 4-(4-(4-(4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of (39) (320 mg, 0.375 mmol) in anhydrous dichloromethane (1.5 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanamine To the 4-thiazolinone (50mg, 0.5mmol) of 4-nitropropane-2-ol (50mL) in the mixture of 4-nitropropane-2-ol (10mL) and 4-nitropropane-2-ol (20mL).Hexafluorophosphate N-oxide (150mg, 0.395mmol) and anhydrous triethylamine (220 μ L, 1.58mmol).Reaction mixture was at room temperature stirred for 30 minutes.Then benzene-1,4-diamine (41mg, 0.38mmol) was added and the gained mixture was at room temperature stirred for 16 hours.Reaction mixture was quenched with a saturated aqueous solution of sodium bicarbonate (20mL) and extracted with dichloromethane (2x50mL).The organic extract merged was washed with the water (30mL) containing a few drops of acetic acid.Then the organic layer was dried over sodium sulfate, filtered and concentrated in a vacuum.Then by column chromatography (silica) purification of the obtained residue, eluted with methanol / dichloromethane (0% to 10%), to obtain title compound (250mg, 71%), it is a milky white solid.

[0692] MS (ES+): m / z = 944 (M+H) + ; LCMS (Method B): t R =3.45min.

[0693] Example 41: (S)-N-(4-aminophenyl)-4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9, 10,12-Hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butane-amido)-1-methyl-1H-pyrrolidone (41)

[0694]

[0695] To (6aS)-3-(4-((5-((4-(5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]-pyrido[1,2-a][1,4]diazepine To a solution of -5(12H)-allyl formate (40) (250 mg, 0.265 mmol) in dichloromethane (3 mL) was added tetrakis(triphenylphosphine)palladium(0) (15 mg, 5 mol%), triphenylphosphine (17 mg, 25 mol%) and pyrrolidine (26 μL, 0.32 mmol). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was subjected to high vacuum for 30 minutes until excess pyrrolidine was thoroughly removed. The resulting residue was then purified by column chromatography (silica) eluting with acetone / dichloromethane (0% to 100%) and then methanol / acetone (0% to 100%) to obtain the title compound (118 mg, 59%) as a yellow solid.

[0696] 1H NMR(DMSO-d6,400MHz)δ9.88-9.96(m,1H),9.81(s,2H),9.50(s,1H),8.32(brs,2H),8.00(d,J=5.7Hz,1H),7.67-7.73(m,2H),7.48(d,J=8.6Hz,2H),7.39(d,J=1.8Hz,1H),7.31-7.35(m,2H),7.30(d,J=1.6Hz,1H),7.27(s,1H),7.22(d,J=1.5Hz,1H),6.96(d,J=1.6Hz,1H),6.80(s,1H),6.51-6.55(m,2H),4.09-4.17(m,1H),3.99-4.05(m,1H),3.90-3.97(m,1H),3.88(s,3H),3.83(s,3H),3.82(s,3H),3.68-3.72(m,1H),3.05-3.16(m,2H),2.44(t,J=7.3Hz,2H),2.02-2.07(m,2H),1.81-1.91(m,1H),1.68-1.78(m,2H),1.56(d,J=4.9Hz,2H); 13 C NMR(DMSO-d6,100MHz)δ168.8,166.3,164.7,159.5,159.2,150.2,147.1,144.7,139.8,137.0,129.6,128.2,126.1,124.6,124.3,122.0,121.8,120.4,120.2,118.8,113.7,111.3,109.6,104.7,67.7,67.2,55.6,51.1,49.2,38.5,36.2,36.1,35.4,31.8,30.2,24.7,23.7,22.5,17.7;MS(ES+):m / z=757(M+H) + ;LCMS(方法A):t R =5.80min。

[0697] Example 42: 5-(4-((tert-Butyloxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)benzo[b] Thiophene-2-carboxylic acid methyl ester (42)

[0698]

[0699] By 4-((tert-butoxycarbonyl) amino)-1-methyl isophthalic acid H-pyrrole-2-formic acid (127mg, 0.530mmol) in N, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (185mg, 0.960mmol) and 4-(dimethylamino) pyridine (147mg, 1.20mmol) in the solution in dimethylformamide (1mL).Reaction mixture was at room temperature stirred 4 hours.Then add 5-aminobenzo [b] thiophene-2-methylformate (100mg, 0.480mmol) and gained mixture was at room temperature stirred 16 hours.Then pour it in frozen water (40mL) and extract with ethyl acetate (3x100mL).The organic extract merged is washed with citric acid aqueous solution (1M, 60mL), sodium bicarbonate saturated aqueous solution (70mL), water (70mL) and salt solution (70mL) sequentially. The organic layer was then dried over sodium sulfate, filtered and concentrated to give the title compound (185 mg, 90%) as a cream-colored solid. The product was carried forward to the next step without any further purification.

[0700] MS (ES+): m / z = 430 (M+H) + ; LCMS (Method B): t R =4.07min.

[0701] Example 43: 5-(4-amino-1-methyl-1H-pyrrole-2-carboxamido)-benzo[b]thiophene-2-carboxylic acid methyl ester Hydrochloride (43)

[0702]

[0703] Methyl 5-(4-((tert-butoxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)benzo-[b]thiophene-2-carboxylate (42) (150 mg, 0.340 mmol) was dissolved in hydrochloric acid (4M in 1,4-dihydrochloric acid). The mixture was added with 4% paraformaldehyde (10% ethanol) (1 mL) and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated in vacuo to give the title compound (118 mg, 95%) as a light brown solid. The product was carried on to the next step without any further purification.

[0704] MS (ES+): m / z = 364 (M+H) + ; LCMS (Method B): t R =2.78min.

[0705] Example 44: (6aS)-2-methoxy-3-(4-((5-((2-(methoxycarbonyl)benzo-[b]thiophen-5-yl)amino (methyl-1H-pyrrol-3-yl)amino)-4-oxo-butoxy)-12-oxo-6-((tetrahydro-2H-pyran- 2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -5(12H)-formene Propyl ester (44)

[0706]

[0707] 4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (238mg, 1.23mmol) and 4-(dimethylamino)pyridine (189mg, 1.55mmol) were added to a solution of 4-(4-amino-1-methyl-1H-pyrroles-2-formamido)benzo[b]thiophene-2-methylformate hydrochloride (43) (225mg, 0.620mmol) in N,N-dimethylformamide (3mL). The reaction mixture was stirred at room temperature for 4 hours. 5-(4-amino-1-methyl-1H-pyrroles-2-formamido)benzo[b]thiophene-2-methylformate hydrochloride (43) (225mg, 0.620mmol) was then added, and the resulting mixture was stirred at room temperature for 16 hours. The mixture was then poured into frozen water (40mL) and extracted with ethyl acetate (3x100mL). The organic extracts merged were washed sequentially with citric acid aqueous solution (1M, 60mL), a saturated aqueous solution of sodium bicarbonate (70mL), water (70mL) and salt solution (70mL). The organic layer was then dried over sodium sulfate, filtered and concentrated in vacuo.The resulting residue was then purified by column chromatography (silica) eluting with acetone / dichloromethane (0% to 30%) to give the title compound (348 mg, 66%) as a brown solid.

[0708] MS (ES+): m / z = 844 (M+H) + ; LCMS (Method B): t R 4.23min.

[0709] Example 45: 5-(4-(4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetramethyl Hydrogen-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]-pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-pyrrole-2-carboxamido)benzo[b]thiophene-2-carboxylic acid (45)

[0710]

[0711] To 4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butanoic acid (44) (327 mg, 0.387 mmol) in 1,4-dimethoxy- To the 4-nitro-1-oxo-2-oxo-4 ...

[0712] MS (ES+): m / z = 831 (M+H) + ; LCMS (Method B): t R =3.82min.

[0713] Example 46: (6aS)-3-(4-((5-((2-((4-aminophenyl)carbamoyl)benzo-[b]thiophene-5- 1-Methyl-1H-pyrrol-3-yl)amino)-4-oxo-butoxy)-2-methoxy-12-oxo-6- ((Tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -5(12H)-Allyl formate (46)

[0714]

[0715] 5-(4-(4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of (45) (50 mg, 0.060 mmol) in anhydrous dichloromethane (1 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanamine To the 4-thiazolinone (5-nitropropane) of 4-nitropropane-2-ol (5-nitropropane-2-ol) in the mixture of 4-nitropropane-2-ol (2-nitropropane-2-ol) and 4-nitropropane-2-ol (2-nitropropane-2-ol) (3-nitropropane-2-ol) was added hexafluorophosphate N-oxide (28mg, 0.072mmol) and anhydrous triethylamine (35 μ L, 0.25mmol).Reaction mixture was at room temperature stirred for 30 minutes.Then benzene-1,4-diamine (7.0mg, 0.066mmol) was added and the gained mixture was at room temperature stirred for 16 hours.Reaction mixture was quenched with a saturated aqueous solution of sodium bicarbonate (20mL) and extracted with dichloromethane (2x50mL).The organic extract merged was washed with the water (30mL) containing a few drops of acetic acid.Then the organic layer was dried over sodium sulfate, filtered and concentrated in a vacuum.Then by column chromatography (silica) purification of the obtained residue, eluted with acetone / dichloromethane (0% to 50%), to obtain title compound (6.8mg, 12%), it is a yellow solid.

[0716] MS (ES+): m / z = 921 (M+H) + ; LCMS (Method B): t R =3.48min

[0717] Example 47: (S)-N-(2-((4-aminophenyl)carbamoyl)benzo[b]thiophen-5-yl)-4-(4-((2- Methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -3-yl)oxy (47)

[0718]

[0719] To (6aS)-3-(4-((5-((2-((4-aminophenyl)carbamoyl)benzo[b]thiophen-5-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]-diazepine To a solution of 5-(12H)-allyl formate (46) (6.8 mg, 0.0074 mmol) in dichloromethane (1 mL) was added tetrakis(triphenylphosphine)palladium(0) (0.4 mg, 5 mol%), triphenylphosphine (0.5 mg, 25 mol%) and pyrrolidine (1 μL, 0.012 mmol). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was subjected to high vacuum for 30 minutes until the excess pyrrolidine was thoroughly removed. The resulting residue was then purified by column chromatography (silica) eluting with acetone / dichloromethane (0% to 100%) followed by methanol / dichloromethane (0% to 5%) to afford the title compound (1.7 mg, 31%) as a light yellow solid.

[0720] 1H NMR (DMSO-d6, 400 MHz) δ 10.13 (s, 1H), 9.98-10.03 (m, 1H), 9.95 (s, 1H), 8.35 8.42 (m, 1H), 8.19 (s, 1H), 8.01 (d, J = 5.7 Hz, 1H), 7.95 (d, J = 8.9 Hz, 1H), 7.33-7.40 (m, 2H), 7.23-7.28 (m, 2H), 7.02 (s, 1H), 6.81 (s, 1H), 6.57 (d, J = 8.7 Hz, 2H), 5.00 (br. s., 2H), 4.10-4.14 (m, 1H), 3.86 (s, 3H), 3.83 (s, 3H), 3.65-3.74 (m, 2H), 3.15-3.19 (m, 1H), 3.06-3.14 (m, 1H), 2.45 (t, J = 7.5 Hz, 3H), 2.11-2.13 (m, 1H), 2.00-2.08 (m, 4H) 1.74 (dd, J = 9.0, 5.3 Hz, 3H); MS (ES+): m / z = 734 (M+H) + ; LCMS (Method A): t R = 5.63 min.

[0721] General synthesis scheme

[0722]

[0723] i) K2CO3, DMF, ethyl 6-bromobutyrate, r.t.; ii) KNO3, TFA, 0-5 °C; iii) KMnO4, acetone, H2O, reflux; iv) HATU, (S)-piperidin-2-ylmethanol, Et3N, CH2Cl2, 0 °C - r.t.; v) H2, Ni / Ra, MeOH; vi) allyl chloroformate, pyridine, CH2Cl2, -10 °C - r.t.; vii) TEMPO, BAIB, CH2Cl, r.t.; viii) pTSA, DHP, EtOAc, r.t.; ix) NaOH, di alkyl, H2O, r.t.; x) RNH2, EDCl, DMAP, DMF, r.t.; xi) PPh3, Pd(PPh3)4, pyrrolidine, CH2Cl2, r.t.

[0724] Example 48: Ethyl 6-(4-formyl-2-methoxyphenoxy)hexanoate (48)

[0725]

[0726]

[0727] A mixture of vanillin (6.5 g, 42.7 mmol), ethyl 6-bromohexanoate (8.0 mL, 45.0 mmol), and potassium carbonate (8.70 g, 63.0 mmol) in N,N-dimethylformamide (50 mL) was stirred at room temperature for 18 h. The reaction mixture was diluted with water (100 mL), separated, and extracted with ethyl acetate (120 mL). The combined organic extracts were sequentially washed with water (100 mL), brine (100 mL), dried over magnesium sulfate, filtered, and concentrated to give the title compound as a light yellow oil (12.5 g, 99%). The product was carried on to the next step without any further purification.

[0728] 1 H NMR (400 MHz, CDC13) δ 9.84 (s, 1H), 7.42-7.44 (dd, J = 8.2, 1.9 Hz, 1H), 7.40 (d, J = 1.9 Hz, 1H), 6.96 (d, J = 8.1 Hz, 1H), 4.08-4.15 (m, 4H), 3.92 (s, 3H), 2.34 (t, J = 7.5 Hz, 2H), 1.87-1.94 (m, 2H), 1.68-1.75 (m, 2H), 1.49-1.56 (m, 2H), 1.25 (t, J = 7.2 Hz, 3H); MS (ES+): m / z = 317 (M+Na) + ; LCMS (Method B): t R = 3.82 min.

[0729] Example 49: Ethyl 6-(4-formyl-2-methoxy-5-nitrophenoxy)hexanoate (49)

[0730]

[0731] To a stirred solution of potassium nitrate (5.4 g, 53 mmol) in trifluoroacetic acid (25 mL) was added dropwise a solution of ethyl 6-(4-formyl-2-methoxyphenoxy)hexanoate (48) (12.5 g, 42 mmol) in trifluoroacetic acid (25 mL) at room temperature. The reaction mixture was stirred for 1 h. It was then concentrated in vacuo and the residue was dissolved in ethyl acetate (200 mL). It was washed with brine (3 x 50 mL) then saturated aqueous sodium bicarbonate solution (2 x 40 mL), dried over magnesium sulfate, filtered, and concentrated in vacuo to give the title compound as a yellow solid (14.4 g, 100%). The product was carried on to the next step without any further purification.

[0732] 1H NMR (400MHz, CDCl3) δ10.43 (s, 1H) 7.58 (s, 1H), 7.40 (s, 1H), 4.10-4.16 (m, 4H), 4.00 (s, 3H), 2.35 (t, J=7.4Hz , 2H), 1.84-1.96 (m, 2H), 1.69-1.76 (m, 2H), 1.50-1.58 (m, 2H), 1.25 (t, J=7.2Hz, 3H); MS (ES+): m / z=340 (M+H) + ; LCMS (Method B): t R =4.02min.

[0733] Example 50: 4-((6-ethoxy-6-oxohexyl)oxy)-5-methoxy-2-nitrobenzoic acid (50)

[0734]

[0735] To a solution of 6-(4-formyl-2-methoxy-5-nitrophenoxy)ethyl hexanoate (49) (7.8g, 23.0mmol) in acetone (200mL) was added a heat (70°C) solution of potassium permanganate (13.6g, 86.0mmol) in water (100ml). The mixture was then stirred at 70°C for 4 hours. The reaction mixture was cooled to room temperature and passed through diatomaceous earth. The cake was then washed with hot water (100mL). A solution of sodium bisulfite in hydrochloric acid (100mL) was added to the filtrate and extracted with dichloromethane (2x200mL). The organic extracts merged were dried over sodium sulfate, filtered and concentrated in vacuo to obtain the title compound as a yellow solid (5.0g, 61%), which was used in subsequent steps without further purification.

[0736] 1 H NMR (400MHz, CDCl3) δ7.34 (s, 1H), 7.14 (s, 1H), 3.96-4.03 (m, 4H), 3.84 (s, 3H), 2.24 (t, J=7.4Hz, 2H) , 1.70-1.77 (m, 2H), 1.55-1.62 (m, 2H), 1.39-1.45 (m, 2H), 1.13 (t, J=7.1Hz, 3H); MS (ES+) m / z=354 (MH) + ; LCMS (Method B): t R =3.63min.

[0737] Example 51: (S)-6-(4-(2-(Hydroxymethyl)piperidine-1-carbonyl)-2-methoxy-5-nitrophenoxy)hexanoic acid Ethyl ester (51)

[0738]

[0739] To a stirred solution of 4-((6-ethoxy-6-oxohexyl)oxy)-5-methoxy-2-nitrobenzoic acid (50) (2.0 g, 5.6 mmol) and trimethylamine (4.70 mL, 33.8 mmol) in dichloromethane (40 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N'-tetramethylurea in one portion. To the 4-thiazolinone (5-nitropropane)-2-ol (1.2g, 4.7mmol) of 4-nitropropane ...

[0740] 1 H NMR (400MHz, CDCl3) δ7.60-7.63 (m, 1H), 6.75-6.77 (m, 1H), 4.02-4.13 (m, 4H), 3.93 (s, 3H), 3.70-3.78 (m, 1H), 3.39-3.68 (m, 1H) , 3.11-3.18 (m, 3H), 2.32 (t, J=7.6Hz, 2H), 1.83-1.91 (m, 2H), 1.39-1.72 (m, 11H), 1.26 (t, J=7.1Hz, 3H); MS (ES+): m / z=453 (M+H) + ; LCMS (Method B): t R =3.63min.

[0741] Example 52: (S)-6-(5-amino-4-(2-(hydroxymethyl)piperidine-1-carbonyl)-2-methoxyphenoxy)hexanoic acid Ethyl ester (52)

[0742]

[0743] To a solution of ethyl (S)-6-(4-(2-(hydroxymethyl)piperidine-1-carbonyl)-2-methoxy-5-nitrophenoxy)hexanoate (51) (1.2 g, 2.7 mmol) in methanol (20 mL) was added - Nickel (slurry in H O) (120 mg). The resulting mixture was hydrogenated at 50 psi in a Parr apparatus for 1.5 hours, then filtered through a pad of Celite and concentrated in vacuo to give the title compound (991 mg, 87%) as a gray oil that solidified upon standing. The resulting material was carried on to the next step without further purification.

[0744] 1 H NMR (400MHz, CDCl3) δ6.69 (s, 1H), 6.32 (s, 1H), 4.13 (m, 4H), 3.98 (t, J=6.5Hz, 2H), 3.79 (s, 3H), 3.67-3.57 (m, 1H), 3.19-3.22 (m, 4H), 2.87 (s, 2H), 2.32-2.36 (m, 2H), 1.82-1.89 (m, 2H), 1.65-1.73 (m, 6H), 1.47-1.55 (m, 3H), 1.27 (t, J=7.1Hz, 3H); MS (ES+): m / z=423 (M+H) + ; LCMS (Method B): t R =3.23min.

[0745] Example 53: (S)-6-(5-(((allyloxy)carbonyl)amino)-4-(2-(hydroxy-methyl)piperidine-1-carbonyl ethyl)-2-methoxyphenoxy)hexanoate (53)

[0746]

[0747] To a solution of (S)-6-(5-amino-4-(2-(hydroxymethyl)piperidine-1-carbonyl)-2-methoxyphenoxy)ethyl hexanoate (52) (1.23 g, 2.91 mmol) and pyridine (542 μL, 6.69 mmol) in anhydrous dichloromethane (20 mL) was added dropwise a solution of allyl chloroformate (278 μL, 2.62 mmol) in dichloromethane (12 mL) at -10°C. The resulting reaction mixture was stirred at room temperature for 0.5 hour, quenched with a saturated aqueous solution of copper (II) sulfate (25 mL), diluted with dichloromethane (10 mL), separated, and washed sequentially with water (20 mL), a saturated aqueous solution of sodium bicarbonate (20 mL), and brine (20 mL). The organic layer was then dried over magnesium sulfate, filtered, and concentrated in vacuo to obtain the title compound (588 mg, 40%) as an orange oil. The resulting material was carried on to the next step without further purification.

[0748] 1H NMR (400MHz, CDCl3) δ8.23 (br s, 1H), 7.70 (br s, 1H), 6.78 (s, 1H), 5.90-6.00 (m, 1H), 5.33-5.38 (m, 1H), 5.24 (dd, J=10.4, 1.3H z, 1H), 4.63 (m, 2H), 4.12 (q, J=7.1Hz, 2H) 4.05 (t, J=6.6Hz, 2H), 3.83 (s, 3H), 3.6 4-3.72(m, 1H), 3.02-3.12(m, 1H), 2.33(t, J=7.6Hz, 2H), 1.84-1.91(m, 2H), 1.67 -1.74 (m, 10H), 1.66-1.54 (m, 4H), 1.26 (t, J=7.1Hz, 3H); MS (ES+): m / z=507 (M+H) + ; LCMS (Method B): t R =3.70min.

[0749] Example 54: (6aS)-3-((6-ethoxy-6-oxohexyl)oxy)-6-hydroxy-2-methoxy-12-oxohexyl 6,6a,7,8,9,10-Hexahydrobenzo[e]pyrido[1,2-a][1,4]-diazepine -5(12H)-allyl formate (54)

[0750]

[0751] To a solution of (S)-ethyl 6-(5-(((allyloxy)carbonyl)amino)-4-(2-(hydroxymethyl)-piperidine-1-carbonyl)-2-methoxyphenoxy)hexanoate (53) (1.7 g, 3.4 mmol) in dichloromethane (80 mL) was added 2,2,6,6-tetramethyl-1-piperidinyloxy (53 mg, 0.30 mmol) and (diacetoxyiodo)benzene (1.3 g, 4.0 mmol). The reaction mixture was stirred at room temperature for 16 hours, then placed in an ice bath and quenched with a saturated aqueous solution of sodium metabisulfite (35 mL). The mixture was diluted with dichloromethane (30 mL), separated, and washed sequentially with a saturated aqueous solution of sodium bicarbonate (30 mL), water (30 mL), and brine (30 mL). The organic layer was then dried over magnesium sulfate, filtered, and concentrated in vacuo. Purification was performed by column chromatography (silica) eluting with ethyl acetate / hexanes (0% to 80%) to give the desired compound (1.1 g, 66%) as a colorless oil.

[0752] 1H NMR (400MHz, CDCl3) δ7.70-7.72 (m, 1H), 7.09-7.13 (m, 1H), 5.80-5.98 (m, 1H), 5.25-5.38 (m, 1H), 5.14-5.19 (m, 2H), 4.63-4.72 (m, 2H), 4.35-4.50 (m, 1H), 4.13 (q, J=7.1Hz, 2H), 4.03-4.08 (m, 1H) , 3.96-4.01(m, 2H), 3.91(s, 3H), 3.81-3.83(m, 1H), 3.45-3.53(m, 1H), 3.03-3.10(m, 1H), 2.33(t , J=7.6Hz, 2H), 1.83-1.90 (m, 2H), 1.62-1.74 (m, 10H), 1.48-1.53 ​​(m, 2H); MS (ES+): m / z=505 (M+H) + ; LCMS (Method B): t R =3.57min.

[0753] Example 55: (6aS)-3-((6-ethoxy-6-oxohexyl)oxy)-2-methoxy-12-oxo-6-((tetra Hydrogen-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido-[1,2-a][1,4]diazepine -5 (12H)-Allyl formate (55)

[0754]

[0755]

[0756] To a mixture containing (6aS)-3-((6-ethoxy-6-oxohexyl)oxy)-6-hydroxy-2-methoxy-12-oxo-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To the solution of -5 (12H)-allyl formate (54) (1.1g, 2.2mmol) in dichloromethane (50mL), add 3,4-dihydro-2H-pyrans (2.00mL, 22.4mmol) and p-toluenesulfonic acid monohydrate (113mg, 1%w / w), and the resulting mixture was stirred at room temperature for 4 hours.Then the reaction mixture was diluted with dichloromethane (50mL), and washed with a saturated aqueous solution of sodium bicarbonate (50mL) and salt solution (50mL). The organic layer was dried over magnesium sulfate, filtered and concentrated to obtain the title compound after eluting with ethyl acetate / hexane (0% to 70%) by column chromatography (silica) purification, as a yellow oil (863mg, 66%).

[0757] 1H NMR (400MHz, CDCl3) δ7.16 (m, 1H), 6.50 (s, 1H), 6.10 (m, 1H), 5.76-5.81 (m, 1H), 5.03-5.14 (m, 2H), 4.57-4.69 (m, 2H), 4.37-4.49 (m, 1H), 4.26-4.34 (m, 1H), 4.12 (q, J=7.1Hz, 2H), 3.94-4.0 1(m, 3H), 3.90(s, 3H), 3.62-3.68(m, 1H), 3.46-3.68(m, 2H), 3.03-3.12(m, 1H), 2.33(t, J=7.4 Hz, 2H), 1.66-1.89 (m, 11H), 1.47-1.57 (m, 6H), 1.25 (t, J=7.1Hz, 3H); MS (ES+): m / z=589 (M+H) + ; LCMS (Method B): t R =4.32min.

[0758] Example 56: 6-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyrrolidone)- pyrido[1,2-a][1,4]diazepine -3-yl) Oxy)hexanoic acid (56)

[0759]

[0760] To (6aS)-3-((6-ethoxy-6-oxohexyl)oxy)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]-diazepine -5(12H)-allylformate (55) (200 mg, 0.34 mmol) in 1,4-dimethoxybenzylformate To the 4-nitro-1-oxo-2-oxo-4 ...

[0761] 1H NMR (400MHz, CDCl3) δ7.18 (s, 1H), 6.19 (s, 1H), 5.99-6.19 (m, 1H), 5.71-5.81 (m, 1H), 5.02-5.12(m, 2H), 4.51-4.67(m, 1H), 4.36-4.48(m, 1H), 4.23-4.31(m, 1H), 3.88-4.00 (m, 7H), 3.46-3.66 (m, 2H), 3.02-3.12 (m, 1H), 2.36 (t, J=7.4Hz, 2H), 1 .79-1.81(m, 2H), 1.65-1.75(m, 10H), 1.49-1.55(m, 7H); MS(ES+): m / z=561(M+H) + ; LCMS (Method B): t R =3.78min.

[0762] Example 57: 4-(4-(4-((tert-Butoxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)- 1-Methyl-1H-pyrrole-2-carboxylic acid methyl ester (57)

[0763]

[0764] To 4-((tert-butoxycarbonyl) amino)-1-methyl isophthalic acid H-pyrrole-2-carboxylic acid (18) (59mg, 0.23mmol) in N, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (67mg, 0.36mmol) and 4-(dimethylamino) pyridine (65mg, 0.53mmol) solution in dimethylformamide (4mL).Reaction mixture was stirred at room temperature for 2 hours.Then 4-(4-aminophenyl)-1-methyl isophthalic acid H-pyrrole-2-methyl formate (41mg, 0.18mmol) was added to reaction mixture, which was then stirred at room temperature for 16 hours.Reaction mixture was poured into frozen water (40mL) and extracted with ethyl acetate (3x100mL).The organic layer merged was washed sequentially with 1M citric acid (60mL), a saturated aqueous solution of sodium bicarbonate (70mL), water (70mL) and salt solution (70mL). The organic layer was dried over sodium sulfate, filtered and concentrated.The resulting residue was purified by column chromatography (silica) eluting with ethyl acetate / dichloromethane (0% to 50%) to give the title compound (36 mg, 45%) as a cream solid.

[0765] MS (ES+): m / z = 453 (M+H) + ; LCMS (Method B): t R =4.07min.

[0766] Example 58: 4-(4-(4-amino-1-methyl-1H-pyrrole-2-carboxamido)-phenyl)-1-methyl-1H-pyrrole Methyl pyrrole-2-carboxylate (58)

[0767]

[0768] Methyl 4-(4-(4-((tert-Butoxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)-phenyl)-1-methyl-1H-pyrrole-2-carboxylate (57) (150 mg, 0.330 mmol) was dissolved in hydrochloric acid (4 M in 1,4-dihydrochloric acid). The mixture was added to 4% paraformaldehyde (10% ethanol) (1 mL) and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated in vacuo to give the title compound (114 mg, 99%) as a brown solid. The product was carried on to the next step without further purification.

[0769] MS (ES+): m / z = 353 (M+H) + ; LCMS (Method B): t R =2.88min.

[0770] Example 59: (6aS)-2-methoxy-3-((6-((5-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrole- 3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-6-oxohexyl)oxy)-12-oxo-6- ((Tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -5(12H)-Allyl formate (59)

[0771]

[0772] 6-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To the product of 4- (4-amino-1-methyl-1H-pyrroles-2-formamido) phenyl) -1-methyl-1H-pyrroles-2-methyl-formiate (58) (150mg, 0.330mmol), the obtained mixture was stirred at room temperature for 16 hours. The mixture was then poured into frozen water (20mL) and extracted with ethyl acetate (3x75mL). The combined organic extracts were washed sequentially with aqueous citric acid solution (1M, 50 mL), saturated aqueous sodium bicarbonate solution (50 mL), water (50 mL) and brine (50 mL). The organic layer was then dried over sodium sulfate, filtered and concentrated in vacuo to obtain the title compound (133 mg, 45%) as a yellow oil. The product was carried on to the next step without further purification.

[0773] MS (ES+): m / z = 896 (M+H) + ; LCMS (Method B): t R =4.25min.

[0774] Example 60: 4-(4-(4-(6-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6- ((Tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]-pyrido[1,2-a][1,4]di Aza -3-yl)oxy)hexanoyl)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2- Formic acid (60)

[0775]

[0776] To (6aS)-2-methoxy-3-((6-((5-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-6-oxohexyl)oxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -5(12H)-allylformate (59) (200 mg, 0.340 mmol) in 1,4-dimethoxybenzylformate To the 4-nitro-1-oxo-2-oxo-4 ...

[0777] MS (ES+): m / z = 882 (M+H) + ; LCMS (Method B): t R =3.92min.

[0778] Example 61: (6aS)-3-((6-((5-((4-(5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrolidone (1-Methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-6-oxohexyl)oxy)-2-methoxy- 12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1, 4] Diazepine -5(12H)-allyl formate (61)

[0779]

[0780] 4-(4-(4-(6-((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of (60) (123 mg, 0.14 mmol) in anhydrous dichloromethane (2 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanamine To the 4-thiazolinone (54mg, 0.14mmol) of hexafluorophosphate N-oxide (54mg, 0.14mmol) and anhydrous triethylamine (117 μ L, 0.84mmol).Reaction mixture was at room temperature stirred for 30 minutes.Then benzene-1,4-diamine (15.1mg, 0.14mmol) was added and the gained mixture was at room temperature stirred for 16 hours.Reaction mixture was quenched with a saturated aqueous solution of sodium bicarbonate (20mL) and extracted with dichloromethane (2x50mL).The organic extract merged was washed with the water (30mL) containing a few drops of acetic acid.Then the organic layer was dried over sodium sulfate, filtered and concentrated in a vacuum.Then by column chromatography (silica) purification of the obtained residue, eluted with acetone / dichloromethane (0% to 50%), to obtain title compound (63mg, 46%), it is a yellow solid.

[0781] MS (ES+): m / z = 972 (M+H) + ; LCMS (Method B): t R =3.55min

[0782] Example 62: (S)-N-(4-aminophenyl)-4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -3-yl)oxy)hexanoyl)-1-methyl-1H-pyrrole-2-carboxamide)phenyl)-1-methyl-1H-pyrrole-2-carboxamide (62)

[0783]

[0784] To (6aS)-3-((6-((5-((4-(5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-6-oxohexyl)oxy)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]-pyrido[1,2-a][1,4]diazepine To a solution of -5(12H)-allyl formate (61) (25 mg, 0.026 mmol) in dichloromethane (1 mL) was added tetrakis(triphenylphosphine)palladium(0) (2.5 mg, 5 mol%), triphenylphosphine (1.7 mg, 25 mol%) and pyrrolidine (21 μL, 0.260 mmol). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was subjected to high vacuum for 30 minutes until excess pyrrolidine was thoroughly removed. The resulting residue was then purified by column chromatography (silica) and eluted with acetate / hexane (0% to 100%) to obtain the title compound (6.8 mg, 33%) as a light yellow solid.

[0785] 1H NMR (DMSO-d6, 400MHz) δ9.81-9.85 (m, 1H), 9.58 (s, 1H), 9.51 (s, 2H), 8.00 (d, J=5.7Hz, 1H), 7.69-7.72 (m, 2H), 7.47-7.49 ( m, 2H), 7.38-7.43 (m, 1H), 7.30-7.35 (m, 2H), 7.18-7.24 (m, 1H), 7.11-7.13 (m, 1H), 7.07 (s, 1H), 6.94-6.98 (m, 1H), 6.80 (br s, 1H), 6.63-6.72 (m, 2H), 6.52-6.54 (m, 1H), 3.95-4.14 (m, 3H), 3.89 (s, 3H), 3.83 (s, 3H), 3.70 (s, 3H), 3.65-3. 69 (m, 1H), 3.17 (d, J = 5.2Hz, 2H), 2.28 (t, J = 6.5Hz, 2H), 1.72-1.78 (m, 4H), 1.62-1.68 (m, 4H), 1.42-1.48 (m, 3H) 13 C NMR (DMSO-d6, 100MHz) δ169.5, 166.3, 164.6, 159.5, 159.2, 150.3, 147.1, 14 4.8, 139.8, 137.0, 129.6, 128.2, 126.6, 124.6, 124.3, 122.7, 122.1, 121.8, 121.7, 120.5, 120.4, 118.7, 113.7, 111.3, 109.6, 109.3, 104.7, 68.1, 55.6, 36.3, 36.1, 35.5, 28.3, 25.2, 25.1, 23.7, 22.5, 17.7; MS (ES+): m / z=785 (M+H) + ; LCMS (Method A): t R =3.08min.

[0786] Example 63: (6S,6aS)-2-methoxy-3-((6-((5-(methoxycarbonyl)-1-methyl-1H-pyrrole-3- 6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10- Hexahydrobenzo[e]pyrido[1,2-a][1,4]-diazepine -5(12H)-Allyl formate (63)

[0787]

[0788] 6-(((6S,6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-6a][1,4]diazepine To a solution of 1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanamine was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanamine To the 4-amino-1-methyl-1H-pyrrole-2-formic acid methyl ester (37mg, 0.24mmol) of hexafluorophosphate N-oxide (76mg, 0.20mmol) and anhydrous triethylamine (115 μ L, 1.14mmol).Reactant mixture was at room temperature stirred for 30 minutes.Then 4-amino-1-methyl-1H-pyrrole-2-formic acid methyl ester (37mg, 0.24mmol) was added, and gained mixture was at room temperature stirred for 16 hours.Reactant mixture was quenched with a saturated aqueous solution of sodium bicarbonate (20mL) and extracted with dichloromethane (2x50mL).The organic extract merged was washed with the water (30mL) containing a few drops of acetic acid.Then the organic layer was dried over sodium sulfate, filtered and concentrated in a vacuum.Then by column chromatography (silica) purification of the obtained residue, eluted with acetone / dichloromethane (0% to 30%), to obtain title compound (82mg, 62%), it is a white solid.

[0789] MS (ES+): m / z = 697 (M+H) + ; LCMS (Method B): t R =3.98min.

[0790] Example 64: 4-(6-(((6S,6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetramethyl Hydrogen-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]-pyrido[1,2-a][1,4]diazepine -3-yl)oxy)hexanamido)-1-methyl-1H-pyrrole-2-carboxylic acid (64)

[0791]

[0792] To (6S,6aS)-2-methoxy-3-((6-((5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)amino)-6-oxohexyl)oxy)-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -5(12H)-allylformate (63) (76 mg, 0.11 mmol) in 1,4-dimethoxybenzylamine To the 4-nitro-1-oxo-2-oxo-4 ...

[0793] MS (ES+): m / z = 683 (M+H) + ; LCMS (Method B): t R =3.68min.

[0794] Example 65: (6S,6aS)-3-((6-((5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrole- 3-yl)amino)-6-oxohexyl)oxy)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6, 6a,7,8,9,10-Hexahydrobenzo[e]-pyrido[1,2-a][1,4]diazepine -5(12H)-Allyl formate (65)

[0795]

[0796] 4-(6-(((6S,6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of (64) (60 mg, 0.090 mmol) in anhydrous dichloromethane (1 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanamine To the 4-thiazolinone (50mg, 1.5mmol) of 4-nitropropane-2-ol (5.0mmol) in 4-nitropropane-2-ol (67.0mg, 0.175mmol) and anhydrous triethylamine (73 μ L, 0.52mmol).Reaction mixture was at room temperature stirred for 30 minutes.Then benzene-1,4-diamine (10mg, 0.10mmol) was added and the gained mixture was at room temperature stirred for 16 hours.Reaction mixture was quenched with a saturated aqueous solution of sodium bicarbonate (20mL) and extracted with dichloromethane (2x50mL).Then the organic extract merged was washed with the water (30mL) containing a few drops of acetic acid.Then the organic layer was dried over sodium sulfate, filtered and concentrated in a vacuum.Then by column chromatography (silica) purification of the obtained residue, eluted with acetone / dichloromethane (30% to 50%+5%MeOH), to obtain title compound (18mg, 26%), it is a brown solid.

[0797] MS (ES+): m / z = 773 (M+H) + ; LCMS (Method B): t R =3.27min.

[0798] Example 66: (S)-N-(4-aminophenyl)-4-(6-(2-methoxy-12-oxo-6a,7,8,9,10,12-hexa Hydrobenzo[e]pyrido[1,2-a][1,4]diazepine -3-yl)oxy)hexane-amido)-1-methyl-1H-pyrrole-2-carboxylic acid Amide (66)

[0799]

[0800] To (6S,6aS)-3-((6-((5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-6-oxohexyl)oxy)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine To a solution of -5(12H)-allyl formate (65) (18 mg, 0.020 mmol) in dichloromethane (1 mL) was added tetrakis(triphenylphosphine)palladium(0) (1.3 mg, 5 mol%) and pyrrolidine (2.3 μL, 0.030 mmol). The reaction mixture was stirred at room temperature for 30 minutes and then subjected to high vacuum for 30 minutes until excess pyrrolidine was thoroughly removed. The resulting residue was then purified by column chromatography (silica) eluting with methanol / dichloromethane (0% to 100%) to afford the title compound (11.6 mg, 86%) as an off-white solid.

[0801] 1 H NMR (DMSO-d6, 400MHz) δ9.78 (s, 1H), 9.48 (s, 1H), 8.00 (d, J = 5.7Hz, 1H), 7.32 (d, J = 8.8Hz, 2H ), 7.25 (s, 1H), 7.17 (d, J = 1.8Hz, 1H), 6.82 (d, J = 1.9Hz, 1H), 6.79 (s, 1H), 6.56 (d, J = 8.6Hz, 2 H), 4.13 (dd, J=5.7, 3.4Hz, 5H), 3.80 (s, 3H), 3.79 (s, 3H), 3.17 (s, 1H), 3.07-3.11 (m, 1H), 2. 26 (t, J=7.2Hz, 3H), 1.75 (dd, J=13.8, 7.0Hz, 6H), 1.60-1.65 (m, 5H); MS (ES+): m / z=587 (M+H) + ; LCMS (Method B): t R=2.72min, MS(ES+): m / z=587(M+H) + ; LCMS (Method A): t R =5.23min.

[0802] Example 67: (6aS)-3-(4-((5-((4-(5-((2-aminoethyl)carbamoyl)-1-methyl-1H-pyrrolidone (1-Methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-2-methoxy-12- Oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4] Diazepines -5(12H)-Allyl formate (67)

[0803]

[0804] 4-(4-(4-(4-(((6aS)-5-((allyloxy)carbonyl)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]-diazepine To a solution of (3-amino-1H-pyrrole-2-carboxylic acid) (270 mg, 0.317 mmol) in anhydrous dichloromethane (6 mL) was added N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanamine To the 4-thiazolinone (140mg, 1.5mmol) of 4-nitropropane-2-ol (50mL) in the mixture of 4-nitropropane-2-ol (1.5mmol) and 4-nitropropane-2-ol (50mL) was added hexafluorophosphate N-oxide (126mg, 0.333mmol) and anhydrous triethylamine (185 μ L, 1.33mmol). The reaction mixture was stirred at room temperature for 30 minutes. Then ethane-1,2-diamine (379mg, 6.33mmol) was added and the resulting mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with a saturated aqueous solution of sodium bicarbonate (20mL) and extracted with dichloromethane (2x50mL). The organic extracts merged were washed with the water (30mL) containing a few drops of acetic acid. The organic layer was then dried over sodium sulfate, filtered and concentrated in a vacuum. The resulting residue was then purified by column chromatography (silica), eluted with ammonia (2M) / dichloromethane (0% to 10%) in methanol, to obtain title compound (180mg, 63%), which was a white solid.

[0805] MS (ES+): m / z = 896 (M+H) + ; LCMS (Method B): t R =3.12min.

[0806] Example 68: (S)-N-(2-aminoethyl)-4-(4-(4-(2-methoxy-12-oxo-6a,7,8,9, 10,12-Hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine -3-yl)oxy)butyramido)-1-methyl-1H-pyrrolidone (1-Methyl-1H-pyrrole-2-carboxamide)phenyl)-1-methyl-1H-pyrrole-2-carboxamide (68)

[0807]

[0808] To (6aS)-3-(4-((5-((4-(5-((2-aminoethyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-2-methoxy-12-oxo-6-((tetrahydro-2H-pyran-2-yl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]-pyrido[1,2-a][1,4]diazepine To a solution of -5 (12H)-allyl formate (67) (22 mg, 0.025 mmol) in dichloromethane (4 mL) was added tetrakis (triphenylphosphine) palladium (0) (1.4 mg, 5 mol%) and pyrrolidine (3.0 μL, 0.037 mmol). The reaction mixture was stirred at room temperature for 2 hours, then subjected to high vacuum for 30 minutes until excess pyrrolidine was thoroughly removed. The resulting residue was then purified by column chromatography (silica) eluted with methanol / dichloromethane (0% to 20%) to obtain the title compound (11 mg, 62%) as a white solid.

[0809] 1H NMR(DMSO-d6,400MHz)δ10.01(s,1H),9.84(s,1H),9.21(br s,2H),8.41(s,1H),8.01(d,J=5.7Hz,1H),7.70(d,J=8.8Hz,2H),7.44(d,J=8.7Hz,2H),7.38(d,J=1.8Hz,1H),7.31(d,J=1.9Hz,1H),7.27(s,1H),7.22(d,J=1.8Hz,1H),6.98(d,J=1.8Hz,1H),6.80(s,1H),4.09-4.19(m,2H),3.99-4.05(m,2H),3.87(s,3H),3.82(m,6H),3.65-3.72(m,2H),3.45-3.50(m,2H),3.16(d,J=5.3Hz,3H),2.96(t,J=5.8Hz,2H),2.45(t,J=7.4Hz,2H),2.00-2.09(m,4H);(DMSO-d6,100MHz)δ203.1,168.8,166.3,164.7,161.6,159.6,150.2,147.1,139.8,137.0,129.5,125.9,124.2,122.0,120.6,120.4,111.2,109.8,109.3,98.8,95.4,85.9,78.8,71.0,67.7,55.6,49.2,48.5,36.3,31.8,30.2,24.7,22.5,17.7;MS(ES+):m / z=709(M+H) + ;LCMS(方法B):t R =2.80min,MS(ES+):m / z=709(M+H) + ;LCMS(方法A):t R =5.38min。

[0810] Example 69: Demonstration of DNA adduct formation by HPLC

[0811] The interaction of C8-linked PDD monomers with double-stranded transcription factor consensus sequences was investigated using an HPLC assay using an X-bridge MS C18 2.5 μM OST column (2.3 x 50 mm) and a gradient of 40% acetonitrile / water and 100 mM TEAB (tetraethylammonium bromide) / water as the mobile phase at a flow rate of 0.5 mL / min and UV detection at 254 nm. A 4:1 molar ratio of ligand:oligonucleotide was used, with each single-stranded oligonucleotide dissolved in 1 M ammonium acetate to form a 1 mM stock solution. The oligonucleotides were first annealed by heating their 1 mM solutions to 70°C for 10 minutes, followed by gradual cooling over 8 hours and storage at -20°C overnight. A 25 μM working solution of the oligonucleotides was then prepared by diluting the annealed stock solution with 100 mM ammonium acetate. The ligands were dissolved in DMSO to form a 10 mM stock solution, which was stored at -20°C for no more than 4 months. A 100 μM working solution of the drug was prepared by diluting the stock solution with 100 mM ammonium acetate. The working solution of the ligand was added to the working solution of the oligonucleotide at room temperature, and the mixture was incubated at room temperature for different time intervals.

[0812] Example 70: Fluorescence Resonance Energy Transfer (FRET) Assay

[0813] Oligonucleotide sequences used for FRET assays were purchased from Eurogentec, Southampton, UK: TAMRA (6-carboxytetramethylrhodamine) and FAM (6-carboxyfluorescein) were the acceptor and donor fluorophores, respectively. Prior to use, a 400 nM solution in FRET buffer (optimized to 50 mM potassium, 50 mM dimethyl cacosate, pH 7.4) was prepared from a 20 μM stock solution. The oligonucleotides were annealed by heating the sample to 90° C. for 10 minutes, then cooling to room temperature and storing at that temperature for 5 hours. FRET buffer was used for dilution from an initial 5 mM DMSO stock solution. Annealed DNA (50 μL) and sample solution (50 μL) were added to each well of a 96-well plate (MJ Research, Waltham, MA) and processed in a DNA Engine Opticon (MJ Research). Fluorescence readings were taken at 0.5° C. intervals from 30-100° C., with a constant temperature of 30 seconds before each reading. The incident radiation was 450-495 nm and detection was performed at 515-545 nm. The raw data were imported into the program Origin (Version 7.0, OringinLab Corp.) and the graph was smoothed using a 10-point moving average and then normalized. Melting temperatures were determined using a script based on the maximum value of the first derivative of the smoothed melting curve. The difference (ΔTm) between the melting temperature of each sample and the melting temperature of the blank was used for comparison purposes.

[0814] Table 1: ΔTm determined after 24 h incubation with double-stranded DNA sequences of transcription factors

[0815]

[0816] Example 71: Cytotoxicity Analysis of C8-Linked PDD Monomers by MTT Assay

[0817] Cell culture

[0818] MDAMB231 (triple negative human breast cancer) was obtained from the American Type Culture Collection. 2 Cell lines were maintained in monolayer culture in flasks (TPP, Switzerland). MDA MB231 cell lines were maintained in high glucose DMEM (4.5 g / l; Invitrogen), fetal bovine serum (10%, Biosera UK), nonessential amino acids (1×; Invitrogen), L-glutamine (2 mM; Invitrogen), and penicillin-streptomycin (1% v / v, Invitrogen). HeLa cell lines were maintained in Dulbecco's Modified Eagle's Medium (DMEM; Invitrogen) supplemented with fetal bovine serum (10% v / v; Invitrogen), L-glutamine (2 mM; Invitrogen), nonessential amino acids (1×; Invitrogen), and penicillin-streptomycin (1% v / v, Invitrogen). For passage, cells are washed with PBS (GIBCO 14040, Invitrogen, UK), incubated with trypsin (GIBCO 25300, Invitrogen, UK), and reseeded in fresh culture medium. For inoculation, cells are counted by microscope (Nikon, USA) using a Neubauer hemocytometer (Assistant, Germany) on a cell non-adherent suspension, which is washed in PBS, digested with trypsin, centrifuged at 8000 rpm for 5 minutes at 8°C, and resuspended in fresh culture medium.

[0819] MTT assay

[0820] Grow cells in normal cell culture conditions using appropriate culture medium at 37°C in a 5% CO2 humidified atmosphere. Adjust the cell count to 10 5Cells / ml were added to each well, and 5,000-20,000 cells were added according to the cell line. Cells were incubated for 24 hours, and 1 μl of the appropriate inhibitor concentration was added to the wells in triplicate. After continuous exposure to each compound for 72 hours, cytotoxicity was determined using brominated 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) (Lancaster Synthesis Ltd, UK) colorimetric assay. [34 The absorbance was quantified spectrophotometrically at λ = 570 nm (Envision Plate Reader, PerkinElmer, USA). The software calculates IC through dose-response analysis 50 value.

[0821] Table 2: IC determined after 72 h exposure to C8-linked PDD monomers 50 Value (nM).

[0822] #imgpt304#

[0823] References

[0824] 1. Antonow, D., and Thurston, DE (2011) Chem Rev 111, 2815-2864.

[0825] 2. Cipolla, L., Araujo, A. C., Airoldi, C., and Bini, D. (2009) Anticancer Agents Med Chem 9, 1-31.

[0826] 3.Gerratana, B. (2012) Med Res Rev 32, 254-293.

[0827] 4.Hartley, JA (2011) Expert Opin Investig Drugs 20, 733-744.

[0828] 5. Kamal, A., Reddy, KL, Devaiah, V., Shankaraiah, N., and Reddy, DR (2006) Mini Rev Med Chem 6, 53-69.

[0829] 6.Hurley,L.H.,Reck,T.,Thurston,D.E.,Langley,D.R.,Holden,K.G.,Hertzberg,R.P.,Hoover,J.R.,Gallagher,G.,Jr.,Faucette,L.F.,Mong,S.M.,(1988)Chem Res Toxicol 1,258-268.

[0830] 7.Wells,G.,Martin,C.R.,Howard,P.W.,Sands,Z.A.,Laughton,C.A.,Tiberghien,A.,Woo,C.K.,Masterson,L.A.,Stephenson,M.J.,Hartley,J.A.,Jenkins,T.C.,Shnyder,S.D.,Loadman,P.M.,Waring,M.J.,和Thurston,D.E.(2006)J Med Chem49,5442-5461.

[0831] 8.Brucoli,F.,Hawkins,R.M.,James,C.H.,Jackson,P.J.,Wells,G.,Jenkins,T.C.,Ellis,T.,Kotecha,M.,Hochhauser,D.,Hartley,J.A.,Howard,P.W.,和Thurston,D.E.(2013)J Med Chem 56,6339-6351.

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[0855] All publications mentioned in the above specification are incorporated herein by reference. Although the illustrative embodiments of the present invention have been disclosed in detail herein with reference to the accompanying drawings, it should be understood that the invention is not limited to the precise embodiments and that various changes and modifications can be made by those skilled in the art without departing from the scope of the invention as defined by the appended claims and their equivalents.

Claims

1. Compounds: (XV) and its salts, in; R1 is H; R2 is H; R3 is selected from H and C 1-12 alkyl; R4 is selected from phenyl, pyrrolyl, N-methylpyrrolyl, furyl, thienyl, imidazolyl, N-methylimidazolyl, benzofuranyl and benzothienyl, optionally substituted with up to three optional substituents selected from OH, C 1-6 Alkyl, CO2R 11 NR 11 R 12 and C(=O)-NH-R 24 ; R 19 It is H; Y1 is CH; Y2 is CH; p is 0 or 1; X1 is O; L is an alkylene chain containing 1 to 12 carbon atoms; X2 is NHC(=O); q is 1; For each A1 group: ; A1 One of Y3 and Y4 is independently selected from NR 17 ; and the other of Y3 and Y4 is CH; and Y5 is independently selected from CH and N; R 24 is phenyl optionally substituted with up to three optional substituents selected from C 1-6 Alkyl, CO2R 11 and NR 11 R 12 ; R 11 、R 12 and R 17 Independently selected from H and C 1-6 alkyl; and R5 and R6 together form a double bond; Provided that when p is 0, then R4 is not pyrrolyl, imidazolyl, optionally substituted pyrrolyl, or optionally substituted imidazolyl.

2. The compound and salt thereof according to claim 1, wherein R 24 Yes-C6H4-R 18 , where R 18 Selected from CO2R 11 and NR 11 R 12 .

3. The compound and salt thereof according to claim 1, wherein R3 is selected from methyl and ethyl.

4. The compound and salt thereof according to claim 1, wherein R4 is selected from: 、 and ; wherein Z1 is selected from NH, N-CH3, S and O; Z2 is selected from CH and N; Z3 is selected from S and O; Z4 is CH; R 22 Selected from CO2R 11 NR 11 R 12 and C(=O)-NH-C6H4-R 18 ; R 18 Selected from CO2R 11 and NR 11 R 12 ;and R 11 and R 12 Independently selected from H and C 1-6 alkyl.

5. The compound and salt thereof according to claim 1, wherein the compound has the following structure: (XVI); (XVII); and (XIX) Where q is 1; p is 0 or 1; L is an alkylene chain containing 1 to 12 carbon atoms; Y1 is CH; Y2 is CH; Y5 is selected from CH and N; Z1 is selected from O, S, NH and N-CH3; Z2 is selected from CH and N; Z3 is selected from S and O; Z4 is CH; R 22 Selected from CO2H, CO2C 1-6 Alkyl, NR 11 R 12 and C(=O)-NH-C6H4-R 18 ; R 18 Selected from CO2R 11 and NR 11 R 12 ; R 19 It is H; R 11 and R 12 Independently selected from H and C 1-6 alkyl; and R5 and R6 together form a double bond; Provided that when the compound is (XVI) and p is 0, then Z1 is selected from O and S.

6. The compound and salt thereof according to claim 1, wherein the compound is selected from: (a) (S)-5-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido-[1,2-a][1,4]diazepin-3-yl)oxy)butyramido)-1-methyl-1H-pyrrole-2-carboxamido)benzo-[b]thiophene-2-carboxylic acid methyl ester (13) (13)； (b) (S)-5-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido-[1,2-a][1,4]diazepin-3-yl)oxy)butyramido)-1-methyl-1H-imidazole-2-carboxamido)-benzo[b]thiophene-2-carboxylic acid methyl ester (17) (17)； (c) (S)-4-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]-pyrido[1,2-a][1,4]diazepin-3-yl)oxy)butyramido)-1-methyl-1H-imidazole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylic acid methyl ester (20) (20)； (d) (S)-4-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]-pyrido[1,2-a][1,4]diazepin-3-yl)oxy)butyramido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylic acid methyl ester (24) (24)； (i) (S)-N-(4-aminophenyl)-4-(4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydro-benzo[e]pyrido[1,2-a][1,4]diazepin-3-yl)oxy)butyric-amido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxamide (41) (41)； and (j) (S)-N-(2-((4-aminophenyl)carbamoyl)benzo[b]thiophen-5-yl)-4-(4-((2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepin-3-yl)oxy)butyramido)-1-methyl-1H-pyrrole-2-carboxamide (47) (47)。 7. A pharmaceutical composition comprising: a compound according to any one of claims 1 to 6 or a salt thereof and a pharmaceutically acceptable excipient, carrier or diluent.

8. Use of the compound according to any one of claims 1 to 6 and a salt thereof in the preparation of a medicament for treating a proliferative disease.

9. The method of claim 8, wherein the proliferative disease is selected from the group consisting of bladder cancer, bone cancer, intestinal cancer, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, esophageal cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, retinoblastoma, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, and uterine cancer.

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