Cancer-treating compounds

Abstract

The present invention relates to a group of compounds generally known as duocarmycins. The present invention provides a novel group of duocarmycins having the structure shown in Formula I. These compounds are effective in the treatment of cancer. JPEG2026506407000088.jpg51170

Description

[Technical Field]

[0001] The present invention relates to a group of compounds generally known as duocarmycins. The present invention relates to these compounds and their use in the treatment of cancer. Additionally, the present invention contemplates methods of treatment using the compounds of the present invention. [Background technology]

[0002] Duocarmycins belong to a class of drugs that have fascinated scientists for over 40 years. Their exceptional activity, unique mechanism of action, and efficacy in multidrug-resistant tumor models have attracted the attention of medicinal chemists and drug explorers. However, despite significant progress in fine-tuning their biological activity through structure-activity relationship studies, their therapeutic potential in cancer treatment remains elusive.

[0003] Duocarmycins are a family of DNA minor groove-binding compounds with exceptional cytotoxicity, originally identified in Streptomyces. Cyclopropapyrroloindole-based duocarmycins, such as (+)-duocarmycin SA (DSA) and CC-1065, consist of a DNA recognition motif (DNA-RM) and a pharmacophore responsible for DNA alkylation. The DNA-RM first binds noncovalently to the minor groove, triggering a conformational change that positions the cyclopropane of the alkylating subunit for nucleophilic attack by the adenine at the N3 position of the AT-rich region. Synthetic manipulation of the DNA alkylating subunit dramatically impacts cellular activity in the pM-nM range, with most compounds inducing apoptosis through direct S-phase inhibition and subsequent cell cycle arrest.

[0004] Derived from common scaffolds such as CI, DSA, CPI, and CBI, Seco-duocarmycins are precursor molecules containing a phenolic hydroxyl group that are responsible for initiating the rearrangement of the pharmacophore to generate cyclopropane-containing cytotoxic agents through a process known as spirocyclization.

[0005] Unfortunately, despite their great therapeutic potential, the duocarmycins have yet to achieve market approval for clinical use. Clinical administration of adozelesin, bizelesin, and two carbamate prodrugs, carzelesin and KW-2189, was associated with severe side effects and narrow therapeutic indices, leading to the halting of all clinical trials. Despite this clinical failure, interest in these molecules remains high, and a vast body of research has uncovered fascinating insights in both biological and chemical exploration. The unique ability of duocarmycins to circumvent traditional resistance mechanisms and maintain exceptional activity in multidrug-resistant cells continues to garner significant interest from both academia and industry to develop new approaches to this group of molecules focused on expanding their therapeutic index for the benefit of patients.

[0006] To help improve the therapeutic index of duocarmycins, several approaches have been investigated, including hypoxia-activated duocarmycin prodrugs, cytochrome P450 oxidation prodrugs, phenol-masking prodrugs, antibody-drug conjugates (ADCs), and small molecule drug conjugates. Each is discussed in detail below.

[0007] Hypoxia-activated duocarmycin prodrugs

[0008] Solid tumors often contain areas of high cell density and poor vascularization, leading to hypoxic conditions and hypoxia. These areas are often treatment-resistant and harbor some of the most aggressive cancer cells. While hypoxic regions present an obstacle to effective treatment, they also present an opportunity for the development of hypoxia-activated prodrugs (HAPs). Typically, HAPs are designed to exploit the presence of one- and two-electron oxidoreductases that can catalyze the reduction of trigger groups critical for prodrug activation and hypoxia selectivity. Initial proof-of-concept (PoC) studies with seco-amino CI-based duocarmycins demonstrated that these compounds possess significant biological activity and are amenable to prodrug derivatization. Through numerous intriguing mechanistic studies, we have demonstrated the hypoxic cytotoxicity ratio (HCR, IC) of 1000 to 10000 cytotoxicity ratios (IC). 50 (oxic) / IC 50 The (hypoxic) CBI analogues were improved, ultimately resulting in the development of sulfonamide-containing CBI analogues suitable for HAP. To improve water solubility for in vivo studies, phosphate preprodrugs were developed. Significant synergy was observed between these two compounds and radiotherapy or chemotherapy (gemcitabine and docetaxel) in mouse xenograph models, with complete remissions observed in some mice. However, due to sporadic acute toxicity, the maximum tolerated dose could not be calculated. While several NO2-containing hypoxia-activated duocarmycin drugs are available on the market, spontaneous bioreduction of the NO2 group can potentially cause off-target effects. Therefore, there is a need for compounds that avoid sporadic acute toxicity events and / or potential off-target events.

[0009] The Boger laboratory described N-acyl O-aminophenol HAPs that were sensitive to cleavage at the weak ON bond, and reported that these compounds were preferentially activated under hypoxic conditions. While early stage compounds had poor in vivo stability, cyclic counterparts were shown to have greater stability, yet still cleaved rapidly under in vitro reducing conditions.

[0010] Eight HAPs have progressed to clinical evaluation, but none have been approved. The future success of duocarmycin-based HAPs is likely to be related to (i) the design of better drug candidates that achieve a balance of enhancing bystander effects while maintaining activity in the tumor microenvironment (TME), and (ii) better clinical trial design in patients with severely hypoxic tumors confirmed by reliable biomarker approaches.

[0011] Cytochrome P450 oxidation

[0012] Cytochrome P450 (CYP) enzymes are a superfamily of mixed-function oxidases, and CYP1-3 subfamily members are unique in their ability to oxidize drugs. Their catalytic activity and substrate specificity have been investigated to assess their potential for drug design. The overexpression of multiple isoforms, including CYP1A1, 1B1, 2S1, and 2W1, in many human tumor types relative to normal tissues presents an opportunity for drug design whose activity relies on key functional groups that can be unmasked or restored by selective CYP oxidation in tumor tissue. In this context, seco-duocarmycins are suitable candidate molecules due to the importance of the phenolic OH group in the central chloromethylindoline trigger unit and the removal of the OH group in the scaffolds of CBI and CPI, which reduces activity by more than 1000-fold. PoC studies have shown that both CYP1A1 and CYP2W1 can restore the phenolic OH group that is critical for biological activity. The lead bioprecursor was studied in vivo and shown to significantly slow tumor growth in a xenograph model expressing CYP1A1 and CYP2W1. Importantly, no weight loss was observed, indicating that these redesigned duocarmycin bioprecursors can be safely administered while maintaining anticancer activity. However, such duocarmycin prodrugs are only applicable to tumors overexpressing the required enzymes. Therefore, a more general approach to providing duocarmycin drug analogs that are tolerated by patients is needed.

[0013] Phenol-masking prodrugs

[0014] Phenol-masking prodrugs are a collection of approaches that share the common feature of masking the phenol (or primary amine) group with a cleavable moiety. Carbamate duocarmycins are an example of phenol-masking, but nonspecific prodrug activation contributed to off-target toxicity.

[0015] Two recent studies investigated seco-CBIs, featuring an O-linked 2-nitropiperonyl UV-labile group via chemical inactivation of the phenolic OH, as photoactivatable therapies. Both studies demonstrated effective in vitro activation after irradiation with 365 nm UV light. A drawback of this therapeutic approach is that the light must be applied to a known tumor mass, which currently limits its use to metastatic cancer. However, photoactivation chemotherapy may be useful for unresectable tumors or when tumor mass removal is incomplete. Therefore, there is a need for duocarmycins that can be activated without the need for exposure to light.

[0016] Attempts have been made to immobilize duocarmycins in the seco form by attaching peptides to the primary amino group of the functional group, making activation dependent on peptide cleavage by tumor-associated peptidases. For example, Twum et al. prepared and investigated aminoCBI for conjugation via the amine to a peptide unit designed for cleavage by prostate-specific antigen (PSA). This work focused on establishing a rigorous chemical synthetic route to generate the appropriate seco-NH2-CBI warhead and confirmed the in vitro cytotoxicity of aminoCBI in PSA-expressing cell lines.

[0017] Extensive research has been conducted on the development of glycoside-modified duocarmycins designed for use in antibody-directed enzyme prodrug therapy (ADEPT) technology. In this system, the phenolic OH group of the DNA-alkylating subunit is chemically modified with an O-linked glycoside group, rendering it inactive by preventing spirocyclization. Cellular activity was demonstrated to be over 1000-fold higher in A549 cells expressing the target glycoside enzyme.

[0018] A study by Tietze, Roffler, and colleagues employed a gene-directed enzyme prodrug therapy (GDEPT) approach. To demonstrate the feasibility of this approach, adenovirus expressing β-glucuronidase fused to an optimized transmembrane domain was administered intratumorally to human CL1-5 xenographs in mice, along with a gene-directed enzyme prodrug. Survival increased from 35 days in untreated mice to over 150 days in mice treated with the prodrug and vector, with 7 of 9 mice achieving complete remission. This experiment demonstrated proof of concept in terms of enzyme delivery using a non-antibody approach and highlighted the feasibility of using delivery options such as bacterial expression systems and nanoparticle delivery technologies.

[0019] Both radiation and chemotherapy treatments have been reported to induce the formation of senescent cells in some cases, which may have tumor growth-suppressing effects but also serve as a means to evade apoptosis, allowing for potential reversal of senescence. Interestingly, based on the higher abundance of β-galactosidase in senescent cells, a galactose-protected bifunctional CBI dimer was recently reported as a potential senolytic. While senolytic function was demonstrated in vitro and in vivo, no toxicity data were presented in this study. However, given the extreme toxicity of duocarmycins and the low expression levels of β-galactosidase in all tissues, systemic toxicity is expected and will require careful management in clinical settings.

[0020] Antibody-drug conjugates

[0021] Antibody-drug conjugates (ADCs) have been gaining attention, with 12 approved drugs and over 100 clinical trials in the past decade. ADCs combine the superior targeting properties of antibodies with the cell-killing power of conjugated payloads.

[0022] Human epidermal growth factor receptor 2 (HER2) is the target of both the clinically approved ADC trastuzumab-emtansine (Kadcyla, T-DM1) and the duocarmycin ADC (SYD985), the latter currently in Phase III clinical trials for breast and gastric cancers. SYD985 uses a duocarmycin-derived payload (duocarmycin-hydroxybenzamide-azaindole, DUBA). DUBA is O-linked to an autolytic group designed to spontaneously decompose after cleavage at the adjacent valine-citrulline (VC) site by cathepsin B. This cleavage releases seco-DUBA, which undergoes spirocyclization to become active. The payload is conjugated to the antibody via a thiol-reactive maleimide to a cysteine ​​group exposed by disulfide bond reduction. The drug-to-antibody ratio (DAR) can be partially controlled by adjusting the amount of reducing agent. The importance of DAR is highlighted by SYD985's precursor, SYD983, which has a wide DAR range from 2 to 8. Separation of the mixture using hydrophobic interaction chromatography (HIC) yielded SYD985, with an average DAR of 2.7, which was twice as effective as SYD983 in killing HER2-expressing SK-BR-3 breast cancer cells.

[0023] A recent study demonstrated that SYD985 is effective against T-DM1-resistant patient-derived xenografts (PDXs). This may be explained by the fact that the SYD985 payload is released without the need for lysosomal degradation, low levels of HER2 are sufficient for cytotoxicity, and the influence of drug efflux pumps is minimal. SYD985 was found to be 3-50 times more effective than T-DM1 in cell lines expressing low levels of HER2, but results were similar when HER2 was expressed at high levels.

[0024] MGC018 is an ADC that uses the same DUBA payload as SYD985 conjugated to a mAb targeting B7-H3, an immune checkpoint antigen highly expressed in most solid tumors. MGC018 recently entered a Phase I / II clinical trial in patients with solid tumors as monotherapy or in combination with MGA012, a PD-1-targeting immune checkpoint inhibitor. Initial safety reports from the trial suggest that side effects are mostly tolerable and show early signs of efficacy in some patients.

[0025] BMS-936561 (MDX-1203), a CD70-targeting ADC containing a duocarmycin as a payload, has previously entered clinical trials. BMS-936561 is conjugated via a linker to a VC cleavage site linked to a DNA-RM of maleimide and duocarmycin. The phenol is protected with a carbamate group that can be removed by endogenous carboxylesterases. However, despite the completion of a phase I clinical trial in renal cell carcinoma and non-Hodgkin's lymphoma and the determination of a well-tolerated dose, the program was terminated.

[0026] A recent study reported on an anti-EGFR bispecific antibody conjugated with a tetra-duocarmycin-cotinine compound. Similar to SYD985, the duocarmycin payload was conjugated via the same O-linked VC autolytic linker, but with a uniform DAR. The conjugate was generated by linking two cotinine modules with the peptide linker GSKGSKGSKGSKK, followed by attaching a chemical toxin to each of the four first lysines. This portion of the conjugate was then conjugated to the cotinine-specific single-chain variable fragment (scVf) of the bispecific antibody. This ADC induced significant tumor growth inhibition in a murine lung adenocarcinoma model, with no detectable weight loss during the 5-week treatment period.

[0027] Additionally, Bondis has four duocarmycin-based ADCs in development.

[0028] As mentioned in the Photoactivation subsection, a therapeutic duocarmycin-antibody conjugate controllable with near-infrared light has been reported. The CBI-based duocarmycin is O-linked to a cyanine caging group via N,N'-diethylethylenediamine at the C4' position and conjugated to panitumumab (an anti-EGFR antibody) via N-hydroxysuccinimide through nonspecific deamidation of asparagine and glutamine. The payload is released upon irradiation with 780 nm light. Furthermore, the conjugate can be visualized with external illumination, which can be used to determine maximum tumor accumulation and therefore the optimal timing of activation. The conjugate exhibited pM-level cytotoxicity in vitro after activation with external light. Reaccumulation is also monitored, facilitating informed second irradiation with activating light. An advantage of this tool is that the low absorption of hemoglobin and water in the 600-850 nm range reduces noise and allows imaging of somewhat deeper tissues.

[0029] Phosphate groups have also been explored for use in ADCs as hydroxy-protecting and water-soluble groups. In general, significantly different cytotoxicity was observed in the presence and absence of phosphorases, indicating that phosphate is preferred in ADC design.

[0030] Small Molecule Drug Conjugates

[0031] Small molecule drug conjugates (SMDCs) are designed to harness the specificity of protein-targeting small molecules and the cytotoxicity of nonspecific chemical toxins by combining them with a cleavable linker.

[0032] An earlier study published in 2014 showed that conjugating an inhibitor of carbonic anhydrase IX (CA9), which is overexpressed in hypoxic cancer cells, with a seco-duocarmycin O-linked to an autolytic group could serve as an effective payload for SMDC. However, the conjugate showed only modest tumor growth inhibition after 7 days of treatment in mice bearing subcutaneously grown SKRC52 ​​renal cell carcinoma.

[0033] More recently, a proof-of-concept study (DUPA-DSA) was reported in which a molecule targeting glutamate carboxypeptidase II (GCPII) was linked to the duocarmycin-derived DSA DNA alkylating subunit via a VC-cleavable linker. GCPII is a cell surface protein that undergoes endocytosis upon ligand binding and is reportedly 1000-fold more abundant in cancer cells, and DUPA-DSA was shown to be significantly more potent in GCPII-expressing cells in vitro.

[0034] Despite the promise of potent cell killing, duocarmycins have yet to be approved for clinical use. Several promising tumor-selective prodrug and ADC approaches have been demonstrated, but none have been relatively successful. Therefore, there remains a need for duocarmycin analogs that can provide effective cancer therapy while avoiding unnecessary side effects. In both prodrug and ADC strategies, low enzyme or antigen expression and tumor infiltration can be obstacles to effective treatment. Furthermore, the exceptional activity of duocarmycins may prevent significant bystander effects and prevent drug concentration decline, which likely contributes to the survival of distinct cell subpopulations. This is not due to resistance, but rather to insufficient drug delivery. Complete eradication of the tumor infrastructure requires further improvement of duocarmycin-based therapies. Summary of the Invention [Means for solving the problem]

[0035] According to the present invention, there is provided a compound of formula I, or a pharmaceutically acceptable salt thereof: [ka] During the ceremony: A represents a 5- or 6-membered ring, wherein there are 3 or 4 atoms not represented within said ring, said 3 or 4 atoms being selected as follows: 0, 1, or 2 of said atoms are independently selected from C=O, N, NH, S, or O, and the remaining atoms of said atoms consist of CH; X represents NH, S or O; Y represents a halo group; R 1 HA-OR 4a , -NR 4a R 4b , substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 5-10 Cycloalkyl, substituted or unsubstituted C 5-10 Heterocycloalkyl, substituted or unsubstituted C 6-10 Aryl, and substituted or unsubstituted C 5-10 heteroaryl, where, if substituted, halo, -OR 4a , -NR 4a R 4b , -NO2, C 1-6 Alkyl, C 1-6 Haloalkyl, C substituted with OH 1-6 Alkyl, OMe-substituted C 1-6 C substituted with alkyl and ═O 1-6 R is one, two, or three groups independently selected from alkyl 1 The group is substituted; R 2 is H, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, and —C(O)—R 5 represents a group selected from: R 3 H, -OH, halo, C 1-6 Alkyl, C1-6 Haloalkyl, C 1-6 Alkoxy, -NR 6a R 6b , C 6-10 Aryl-substituted C 1-6 Alkoxy, and C 6-10 Aryl-substituted C 1-6 alkyl; R 4a and R 4b are each independently, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C substituted with OH 1-6 Alkyl, OMe-substituted C 1-6 Alkyl, C substituted with =O 1-6 Alkyl, substituted or unsubstituted C 6-10 Aryl-substituted C 1-6 Alkyl, -C(O)-C 1-6 Alkyl, -(CR 7a R 7b ) n C(O)OR 8 , -(CR 7a R 7b ) n C(O)NHR 8 , substituted or unsubstituted C 5-10 Cycloalkyl, substituted or unsubstituted C 5-10 Heterocycloalkyl, substituted or unsubstituted C 6-10 Aryl, and substituted or unsubstituted C 5-10 heteroaryl; or R 4b is H; where, if substituted, R 4a Groups and R 4b Groups include halo, OH, -NO2, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C substituted with OH 1-6 Alkyl, OMe-substituted C 1-6 Alkyl, C substituted with =O 1-6 substituted with one or two groups selected from alkyl, and -NH; n is 1, 2, or 3 (preferably 1); R 5 is C 1-6 Alkyl, C 2-6 Alkenyl, and C 2-6 alkynyl; R 6a and R 6b are each independently H and C 1-6 alkyl; R 7a and R 7b One of them is H, C 1-6 Alkyl and -OH substituted C 1-6 Alkyl, -SH, -SMe, -NH2, -C(O)OH, -C(O)NH2, guanidine, C 6-10 Aryl, C substituted with -OH 6-10 Aryl, or C 5-10 heteroaryl; and all other R 7a and R 7b is H; and R 8 is H and C 1-6 alkyl; However, R 2 is methyl, R 1 is not -OBn.

[0036] According to the present invention, there is provided a compound of formula I, or a pharmaceutically acceptable salt thereof: JPEG2026506407000003.jpg52170 formula: A represents a 5- or 6-membered ring, wherein there are 3 or 4 atoms not represented within said ring, said 3 or 4 atoms being selected as follows: 0, 1, or 2 of said atoms are independently selected from C=O, N, NH, or O, and the remaining atoms of said atoms consist of CH; R 1 HA-OR 4a , -NR 4a R 4b , substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 5-10 Cycloalkyl, substituted or unsubstituted C5-10 Heterocycloalkyl, substituted or unsubstituted C 6-10 Aryl, and substituted or unsubstituted C 5-10 heteroaryl, where, if substituted, halo, -OR 4a , -NR 4a R 4b , C 1-6 Alkyl, C 1-6 Haloalkyl, C substituted with OH 1-6 Alkyl, OMe-substituted C 1-6 C substituted with alkyl and ═O 1-6 R is one, two, or three groups independently selected from alkyl 1 The group is substituted; R 2 is H, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, and —C(O)—R 5 represents a group selected from: R 3 H, -OH, halo, C 1-6 Haloalkyl, C 1-6 Alkoxy, -NR 6a R 6b , C 6-10 Aryl-substituted C 1-6 Alkoxy, and C 6-10 Aryl-substituted C 1-6 alkyl; R 4a and R 4b are independently H, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C substituted with OH 1-6 Alkyl, OMe-substituted C 1-6 Alkyl, C substituted with =O 1-6 Alkyl, C 6-10 Aryl-substituted C 1-6 Alkyl, -C(O)-C 1-6 Alkyl, -(CR 7a R 7b ) n C(O)OR 8, -(CR 7a R 7b ) n C(O)NHR 8 , substituted or unsubstituted C 5-10 Cycloalkyl, substituted or unsubstituted C 5-10 Heterocycloalkyl, substituted or unsubstituted C 6-10 Aryl, and substituted or unsubstituted C 5-10 heteroaryl; where, if substituted, R 4a Groups and R 4b Groups include halo, OH, -NO2, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C substituted with OH 1-6 Alkyl, OMe substituted C 1-6 Alkyl, C substituted with =O 1-6 substituted with one or two groups selected from alkyl, and -NH; n is 1, 2, or 3 (preferably 1); R 5 is C 1-6 Alkyl, C 2-6 Alkenyl, and C 2-6 alkynyl; R 6a and R 6b are each independently H and C 1-6 alkyl; R 7a and R 7b One of them is H, C 1-6 Alkyl and -OH substituted C 1-6 Alkyl, -SH, -SMe, -NH2, -C(O)OH, -C(O)NH2, guanidine, C 6-10 Aryl, C substituted with -OH 6-10 Aryl, or C 5-10 heteroaryl; and all other R 7a and R 7b is H; and R 8 is H and C 1-6 alkyl; However, R 2 is methyl, R 1 is not -OH.

[0037] In certain embodiments, A represents a 5- or 6-membered ring, wherein there are 3 or 4 atoms not represented within said ring, said 3 or 4 atoms being selected as follows: 0, 1, or 2 of said atoms are independently selected from C═O, N, NH, or O, and the remaining atoms of said atoms consist of CH;

[0038] In an embodiment, X is NH.

[0039] In an embodiment, Y is chloro or bromo.

[0040] For the avoidance of doubt, the ring represented by A includes the two carbon atoms forming either end of the circular line representing A, totaling five or six atoms. Thus, the three or four atoms of A plus the two carbon atoms at either end totals a five- or six-membered ring. The five- or six-membered ring represented by A may be aromatic.

[0041] A may represent one of the following rings: benzene, pyridine, pyridazine, pyrimidine, pyrazine, pyrrole, pyrazole, furan, pyrrolinone, imidazole, or oxazole.

[0042] A may represent one of the following rings: thiophene, benzene, pyridine, pyridazine, pyrimidine, pyrazine, pyrrole, pyrazole, furan, pyrrolinone, imidazole, or oxazole.

[0043] In embodiments, the compound of formula I is a compound represented by one of the following formulas: [ka]

[0044] In certain embodiments, in compounds of formulas IA through IL, X is NH.

[0045] Preferably, the compounds of the invention are of formula IA. [ka]

[0046] Preferably, the compound of the invention is of formula IA'. [ka]

[0047] In an embodiment of the present invention, -OR 2 The group may be substituted at any suitable carbon atom of the indole ring, including the nitrogen-containing ring, and the bond enters the carbon atom of the indole ring by replacing the hydrogen atom that is normally substituted at that carbon atom. In an embodiment, the compound of Formula I is a compound shown in Formula IAa: [ka]

[0048] R 1 Groups and R 3 Groups may be substituted at carbon atoms on the ring, usually by replacing a hydrogen atom substituted at that carbon atom, to which a bond is attached. As representative examples of compounds of formula (I), we have shown possible substitution positions on the ring of compounds of formulas IA and IB. R 1 R may be substituted at position a or b as shown below. 3 may be substituted at positions c, d, e, or f. In certain embodiments, R 1 is substituted at position b. In certain embodiments, R 3 is replaced at position f. [ka]

[0049] In an embodiment, the compound of formula I is a compound shown in formula IIA: [ka]

[0050] In certain embodiments, R 3 H, -OH, halo, C 1-6 Haloalkyl, C 1-6 Alkoxy, -NR 6a R 6b , and C 6-10 Aryl-substituted C 1-6 represents a group selected from alkoxy.

[0051] In certain embodiments, R 3 are H, -OH, C 1-6 Alkyl, C 1-6 Alkoxy, -NR 6a R 6b , and C 6-10 Aryl-substituted C 1-6 represents a group selected from alkoxy.

[0052] In certain embodiments, R 6a and R 6b are each independently selected from H, methyl, and ethyl.

[0053] In certain embodiments, R 3 represents a group selected from H, —OH, methyl, ethyl, —OMe, —NH, —NHMe, —N(Me) and —OBn. 3 is H or methyl.

[0054] Thus, in an embodiment, the compound of formula I is a compound shown in formula IIa: [ka]

[0055] In certain embodiments, R 2 is C 1-6Alkyl and -C(O)-R 5 represents a group selected from

[0056] In certain embodiments, R 2 is H, methyl, ethyl, propyl, butyl, and -C(O)-R 5 Preferably, R 2 is methyl or -C(O)-R 5 is.

[0057] R 5 is C 1-6 Alkyl or C 2-6 In certain embodiments, R 5 is methyl or pentynyl.

[0058] Thus, in certain embodiments, R 2 is methyl, —C(O)-methyl, or —C(O)-pentynyl (optionally, the pentynyl has a terminal triple bond). 2 is methyl, -C(O)-methyl, or [ka] may be.

[0059] Thus, the compound of the present invention may be a compound of formula IIIa or IIIb. [ka]

[0060] In certain embodiments, the compound of the invention is a compound of formula IVa or IVb: [ka]

[0061] As mentioned above, in the embodiment, R 3is preferably H. Thus, the compound of the present invention may be a compound shown in formula Va or Vb: [ka]

[0062] In certain embodiments, the compound of the present invention may be a compound shown in Formula VI: [ka]

[0063] In certain embodiments, the compound of the present invention may be a compound shown in Formula VIIa or VIIb. [ka]

[0064] In embodiments, the compound of the present invention may be a compound of formula VIIIa or VIIIb: [ka]

[0065] R 1 -OR 4a , -NR 4a R 4b , substituted or unsubstituted C 5-10 Heterocycloalkyl, substituted or unsubstituted C 6-10 Aryl, and substituted or unsubstituted C 5-10 heteroaryl; where, if substituted, halo, -OR 4a , -NR 4a R 4b , -NO2, C 1-6 Alkyl, C substituted with OH 1-6 C substituted with alkyl and ═O 1-6 R is one, two, or three groups (optionally one or two groups) independently selected from alkyl 1 The group is substituted.

[0066] In embodiments, R 1 is not -OBn.

[0067] R 1 -OR 4a , -NR 4a R 4b , unsubstituted C 5-10 Heterocycloalkyl, substituted or unsubstituted C 6-10 Aryl and substituted or unsubstituted C 5-10 It may represent a group selected from heteroaryl. where, if substituted, halo, -OR 4a , -NR 4a R 4b , C 1-6 Alkyl, C substituted with OH 1-6 C substituted with alkyl and ═O 1-6 R is one or two groups independently selected from alkyl 1 The group is substituted.

[0068] In certain embodiments, R 1 If is substituted, R 1 is substituted with one, two, or three groups (optionally one or two groups) selected from -OH, -OMe, -CHOH, -CHOMe, -NO, -Me, -NH, -NMe, -OH, -F, -C(O)CH, and -OC(O)CH.

[0069] R 4a and R 4b are each independently, C 1-6 Alkyl, C substituted with OH 1-6 Alkyl, OMe-substituted C 1-6 Alkyl, C 6-10 Aryl-substituted C 1-6 Alkyl, -(CR 7a R 7b ) n C(O)OR 8 , -(CR 7a R 7b ) nC(O)NHR 8 , substituted or unsubstituted C 5-10 Cycloalkyl, substituted or unsubstituted C 5-10 Heterocycloalkyl, and substituted or unsubstituted C 6-10 aryl; or R 4b may be H; where, if substituted, R 4a Groups and R 4b Groups include halo, OH, -NO2, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C substituted with OH 1-6 Alkyl, OMe-substituted C 1-6 Alkyl, C substituted with =O 1-6 alkyl, and -NH2 (optionally halo, -NO2, C 1-6 Alkyl, C 1-6 Alkoxy, and C 1-6 haloalkyl).

[0070] In certain embodiments, R 4a and R 4b are each independently, C 1-6 Alkyl, C substituted with OH 1-6 Alkyl, C 6-10 Aryl-substituted C 1-4 Alkyl, -(CR 7a R 7b )C(O)OR 8 , -(CR 7a R 7b )C(O)NHR 8 , unsubstituted C 5-10 Cycloalkyl, unsubstituted C 5-10 Heterocycloalkyl, and substituted or unsubstituted C 6-10 aryl; or R 4b may be H; where, if substituted, R 4a Groups and R 4b Groups include halo, OH, -NO2, C 1-6 Alkyl, C 1-6Alkoxy, C 1-6 Haloalkyl, C substituted with OH 1-6 Alkyl, OMe-substituted C 1-6 Alkyl, C substituted with =O 1-6 It is substituted with one or two groups (optionally one group) selected from alkyl, and -NH2.

[0071] In certain embodiments, R 4a is C 1-6 Alkyl, C substituted with OH 1-6 Alkyl, C 6-10 Aryl-substituted C 1-4 Alkyl, -(CR 7a R 7b )C(O)OR 8 , -(CR 7a R 7b )C(O)NHR 8 , unsubstituted C 5-10 Cycloalkyl, unsubstituted C 5-10 Heterocycloalkyl, and substituted or unsubstituted C 6-10 aryl; and R 4b is H or C 1-6 alkyl; where, if substituted, R 4a Groups include halo, OH, -NO2, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C substituted with OH 1-6 Alkyl, OMe-substituted C 1-6 Alkyl, C substituted with =O 1-6 It is substituted with one or two groups (optionally one group) selected from alkyl, and -NH2.

[0072] If substituted, R 4a Groups and R 4b Groups include halo, -NO2, C 1-6 Alkyl, C 1-6 Alkoxy, and C 1-6 It may be substituted with one or two groups (optionally one group) selected from haloalkyl.

[0073] In certain embodiments, R 4a is C 1-6 Alkyl, C substituted with OH 1-6 Alkyl, C 6-10 Aryl-substituted C 1-4 Alkyl, -(CR 7a R 7b )C(O)OR 8 , -(CR 7a R 7b )C(O)NHR 8 , unsubstituted C 5-10 Cycloalkyl, unsubstituted C 5-10 Heterocycloalkyl, and substituted or unsubstituted C 6-10 aryl; and R 4b is H or C 1-6 alkyl; where, if substituted, R 4a Groups include halo, -NO2, C 1-6 Alkyl, C 1-6 Alkoxy, and C 1-6 It is substituted with one or two groups (optionally one group) selected from haloalkyl.

[0074] R 4a and R 4b may be substituted with one or two groups preferably selected from -OMe, -CF3, -Cl, -NO2, and -Me.

[0075] In certain embodiments, R 7a and R 7b One of them is H, C 1-6 Alkyl and C substituted with -OH, -SH, -SMe, -NH2, -C(O)OH, -C(O)NH2, guanidine, benzene, phenol, indole or imidazole 1-6 alkyl, and all other R 7a and R 7b is H.

[0076] In certain embodiments, n is 1 and R 7a and R7b One of them is C 1-6 Alkyl and C substituted with -OH, -SH, -SMe, -NH2, -C(O)OH, -C(O)NH2, guanidine, benzene, phenol, indole or imidazole 1-6 alkyl, and R 7a and R 7b The other of these is H.

[0077] In embodiments, R 7a and R 7b One of them is R 7a and R 7b and R is an amino acid side chain that meets the definition of any of the preceding definitions. 7a and R 7b may be -CH2OH, -CH(OH)CH3, -CH2C(O)NH2, CH2CH2C(O)NH2, -CH2SH, -CH2CH2SCH3, -CH2NH2, -(CH2)4NH2, -CH2C(O)OH, -CH2CH2C(O)OH, methyl, propyl, butyl, benzyl, -CH2-phenolyl, -CH2-indolyl, -CH2-imidazolyl, -CH2CH2CH2-guanidinyl, and all other R 7a and R 7b is H.

[0078] In certain embodiments, R 8 is H or Me.

[0079] In certain embodiments, R 1 is OBn (optionally, R 2 is not Me), -N(Et)2, -NHBu, and: [ka] is selected from.

[0080] In certain embodiments, R 1 is -OH (optionally, R 2 is not H), OBn, and: [ka] is selected from.

[0081] In one embodiment of the present invention, the compound of formula I is selected from: [ka] [ka] [ka] [ka] [ka] [ka]

[0082] In certain embodiments, the compound of formula I is selected from: [ka] [ka]

[0083] In another aspect of the invention, there is provided a compound of formula I for use as a pharmaceutical.

[0084] In another embodiment, the compounds of formula I are for use in the treatment of a condition treatable by administration of a cytotoxic agent.

[0085] In another embodiment, the compound of formula I is for use in the treatment of cancer.

[0086] In another embodiment, a method for treating a condition treatable by administration of a cytotoxic agent is provided, the method comprising administering a pharmacologically effective amount of a compound of formula I.

[0087] In another embodiment, a method of treating cancer is provided, the method comprising administering a pharmacologically effective amount of a compound of formula I.

[0088] In certain embodiments, the condition treatable by administration of a cytotoxic agent is cancer. The cancer treatable in any aspect or embodiment of the present invention may be a solid tumor, lymphoma, or leukemia. The solid tumor may be selected from, but is not limited to, breast, lung, prostate, colon, bladder, brain, pancreatic, head and neck, and neuroblastoma cancers. In certain embodiments, the cancer is breast cancer.

[0089] In another aspect of the present invention, compounds of the present invention suitable for conjugation with antibodies are provided. Here, for simplicity in discussing drug conjugates, the term "antibody" includes the conventional meaning of antibody, as well as biosimilars, affimers, or other non-antibody binding proteins. Accordingly, the present invention provides the use of compounds of the present invention in the manufacture of antibody-drug conjugates (meaning antibodies, biosimilars, affimers, or other non-antibody binding protein drug conjugates). Antibody-drug conjugates can be prepared by substituting a compound of the present invention with a linker group and attaching an antibody to the linker group, or by substituting a compound of the present invention with a linker-antibody conjugate.

[0090] In one aspect of the invention, an antibody drug conjugate is provided, wherein the drug is a compound of the invention. In another aspect of the invention, an antibody drug conjugate is provided, comprising a compound of Formula I, an antibody, a biosimilar, an affimer, or other non-antibody binding protein, and a linker connecting the compound of Formula I to the antibody.

[0091] In certain embodiments, the linker is at position 4, 5, 6, or 7 as shown in the structure below, or at R 1The group is attached to the compound of formula I at any chemically possible position on the group. [ka]

[0092] Any linker known in the art is suitable for use in the present invention. The linker must be suitable for linking to a compound of Formula I and to an antibody. Therefore, linkers are generally bifunctional, having a first functional group at a first end of the molecule and a second functional group at a second end of the molecule. The first and / or second functional groups of the linker can be ethers, amines, esters, carbonates, carbamates, thioethers, or other sulfur-bridging units, or triazoles formed via click chemistry. The linker molecule can comprise an alkyl chain, an alkenyl chain, an alkynyl chain, polyethylene glycol (or other similar moieties for improving water solubility), or a peptide. The alkyl chain, alkenyl chain, or alkynyl chain can be of any length, optionally from 3 to 10 atoms long. The polyethylene glycol can consist of 2 to 10 repeating units. The peptide can consist of 1 to 6 amino acid residues.

[0093] Linkers may include autolytic modules, cleavable peptides, and antibody-binding units. These characteristics of linkers within the context of antibody-drug conjugates are well established and known to those skilled in the art.

[0094] The linker may be cleavable (eg, by enzymatic or chemical means) or non-cleavable.

[0095] For example, the use of antibody-drug conjugates with linkers contemplated in the present invention is discussed in Yao et al, Drug Discovery Today, 2021, 26(8), 1857-1874; Jukes et al, Drug Discovery Today, 2021, 26(2), 577-584; Khongorzul et al., 2020, 18(1), 3-19; Dumontet, Nat Revs Drug Discov, 2023, 22, 641-661; Dokter et al, Mol. Cancer Ther. 2014, 13(11), 2618-2629; and van der Lee, Mol Cancer Ther, 2015, 14(3), 692-703, all of which are incorporated herein by reference.

[0096] It may be suitable for the present application and is also discussed in US 7,964,566 and US 2017 / 0232108, both of which are incorporated by reference. [Brief explanation of the drawings]

[0097] Embodiments of the invention are further described below with reference to the accompanying drawings, in which: [Figure 1] FIG. 1 is a Western blot showing phosphorylation of H2AX as a marker of double-stranded DNA damage. [Figure 2] FIG. 2 is a Western blot showing the expression of phosphorylated γ-H2AX as a marker of DNA damage in MCF-7 cells after treatment with duocarmycin compounds. DETAILED DESCRIPTION OF THE INVENTION

[0098] The following are definitions of terms used in this application: Any term not defined herein shall have the ordinary meaning as understood by one of ordinary skill in the art.

[0099] The term "halo" refers to one of the halogens in Group 17 of the periodic table. In particular, the term refers to fluorine, chlorine, bromine, and iodine. Preferably, the term refers to fluorine or chlorine.

[0100] "C 1-6 The term "alkyl" refers to a straight or branched hydrocarbon chain containing 1, 2, 3, 4, 5, or 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, and n-hexyl. Alkyl groups can be unsubstituted or substituted with one or more substituents. Examples of substituents are described below. Substituents for alkyl groups include halogens (e.g., fluorine, chlorine, bromine, and iodine), OH, C 1-6 It may be alkoxy.

[0101] "C 1-6 The term "alkoxy" refers to an alkyl group attached to a molecule via oxygen. It includes moieties (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, and n-hexyl) where the alkyl portion can be straight or branched and can contain 1, 2, 3, 4, 5, or 6 carbon atoms. Thus, alkoxy groups can be methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, and n-hexoxy. The alkyl portion of an alkoxy group can be unsubstituted or substituted with one or more substituents. Examples of substituents are described below. Substituents for alkyl groups include halogens (e.g., fluorine, chlorine, bromine, and iodine), OH, C 1-6 It may be alkoxy.

[0102] "C 1-6 The term "haloalkyl" refers to a hydrocarbon chain substituted with at least one independently selected halogen atom (e.g., fluorine, chlorine, bromine, and iodine) at each occurrence. The halogen atom can be located at any position on the hydrocarbon chain. For example, C 1-6Haloalkyl may refer to chloromethyl, fluoromethyl, trifluoromethyl, chloroethyl (e.g., 1-chloromethyl and 2-chloroethyl), trichloroethyl (e.g., 1,2,2-trichloroethyl), 2,2,2-trichloroethyl, fluoroethyl (e.g., 1-fluoromethyl and 2-fluoroethyl), trifluoroethyl (e.g., 1,2,2-trifluoroethyl and 2,2,2-trifluoroethyl), chloropropyl, trichloropropyl, fluoropropyl, or trifluoropropyl.

[0103] "C 2-6 The term "alkenyl" refers to a branched or straight hydrocarbon chain containing at least one double bond and having 2, 3, 4, 5, or 6 carbon atoms. The double bond may exist as an E or Z isomer. The double bond may be in any possible position on the hydrocarbon chain. For example, "C 2-6 "Alkenyl" can be ethenyl, propenyl, butenyl, butadienyl, pentenyl, pentadienyl, hexenyl and hexadienyl.

[0104] "C 2-6 The term "alkynyl" refers to a branched or straight hydrocarbon chain containing at least one triple bond and having 2, 3, 4, 5, or 6 carbon atoms. The triple bond may be located at any available position on the hydrocarbon chain. For example, "C 2-6 "Alkynyl" can be ethynyl, propynyl, butynyl, pentynyl, and hexynyl.

[0105] The term "carbocyclic" refers to a saturated or unsaturated carbon-containing ring system. A "carbocyclic" system may be monocyclic or a fused polycyclic ring system, e.g., bicyclic or tricyclic. A "carbocyclic" moiety may contain 3 to 14 carbon atoms, e.g., 3 to 8 carbon atoms in a monocyclic system and 7 to 14 carbon atoms in a polycyclic system. "Carbocyclic" encompasses fused ring systems, including cycloalkyl moieties, cycloalkenyl moieties, aryl ring systems, and aromatic moieties.

[0106] The term "heterocyclic" refers to a saturated or unsaturated ring system containing at least one heteroatom selected from N, O, or S. A "heterocyclic" system may contain 1, 2, 3, or 4 heteroatoms, e.g., 1 or 2. A "heterocyclic" system may be monocyclic or a fused polycyclic ring system, e.g., bicyclic or tricyclic. A "heterocyclic" moiety may contain 3 to 14 carbon atoms, e.g., 3 to 8 carbon atoms in a monocyclic system and 7 to 14 carbon atoms in a polycyclic system. "Heterocyclic" encompasses heterocycloalkyl, heterocycloalkenyl, and heteroaromatic moieties. For example, the heterocyclic group can be oxirane, aziridine, azetidine, oxetane, tetrahydrofuran, pyrrolidine, imidazolidine, succinimide, pyrazolidine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, piperidine, morpholine, thiomorpholine, piperazine, and tetrahydropyran.

[0107] The term "cycloalkyl" refers to a saturated hydrocarbon ring system. A "cycloalkyl" group may also be designated as "C3-10 cycloalkyl," which contains 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. The ring system may be a single ring, a bicyclic, or a tricyclic ring system. For example, a "cycloalkyl" may be cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclohexyl, cycloheptyl, and cyclooctyl.

[0108] The term "cycloalkenyl" refers to an unsaturated hydrocarbon ring system that is not aromatic. A "cycloalkenyl" group is defined as "C 3-10 It may be written as "cycloalkenyl". 3-10"Cycloalkenyl" is a ring system containing 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. The ring may contain more than one double bond, provided the ring system is not aromatic. The ring system may be a single ring, or a bicyclic or tricyclic ring system. For example, "cycloalkenyl" can be cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl, cycloheptadiene, cyclooctenyl, and cycloatadienyl.

[0109] The term "heterocycloalkyl" refers to a saturated hydrocarbon ring system having at least one heteroatom in the ring selected from N, O, and S. A "heterocycloalkyl" group is defined as "C 3-10 It may be written as "heterocycloalkyl". 3-10 "Heterocycloalkyl" refers to a ring system containing 3, 4, 5, 6, 7, 8, 9, or 10 atoms, at least one of which is a heteroatom. For example, there may be 1, 2, or 3 heteroatoms, optionally 1 or 2. A "heterocycloalkyl" group may also be referred to as a "3- to 10-membered heterocycloalkyl," which is also a ring system containing 3, 4, 5, 6, 7, 8, 9, or 10 atoms, at least one of which is a heteroatom. The ring system may be a single ring, or a bicyclic or tricyclic ring system. If the ring system is bicyclic, one of the rings may be aromatic, as in, for example, in indane. A "heterocycloalkyl" may be attached to the rest of the molecule through any carbon atom or heteroatom. A "heterocycloalkyl" may have one or more (e.g., one or two) bonds to the rest of the molecule; these bonds may be through any of the atoms in the ring. For example, a "heterocycloalkyl" can be oxirane, aziridine, azetidine, oxetane, tetrahydrofuran, pyrrolidine, imidazolidine, succinimide, pyrazolidine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, piperidine, morpholine, thiomorpholine, piperazine, tetrahydropyran, and indane.

[0110] The term "heterocycloalkenyl" refers to a non-aromatic unsaturated hydrocarbon ring system having at least one heteroatom selected from N, O, and S within the ring. A "heterocycloalkenyl" group is defined as "C 3-10 It may also be written as "heterocycloalkenyl". 3-10 A "heterocycloalkenyl" is a ring system containing 3, 4, 5, 6, 7, 8, 9, or 10 atoms, at least one of which is a heteroatom. For example, there may be 1, 2, or 3 heteroatoms, optionally 1 or 2. A "heterocycloalkenyl" group may also be referred to as a "3- to 10-membered heterocycloalkenyl," which is also a ring system containing 3, 4, 5, 6, 7, 8, 9, or 10 atoms, at least one of which is a heteroatom. The ring system may be a single ring, or a bicyclic or tricyclic ring system. If the ring system is bicyclic, one of the rings may be aromatic, as in, for example, indoline and dihydrobenzofuran. A "heterocycloalkenyl" may be bonded to the rest of the molecule through any carbon atom or heteroatom. A "heterocycloalkenyl" may have one or more (e.g., one or two) bonds to the rest of the molecule; these bonds may be through any of the atoms in the ring. For example, "C 3-8 The "heterocycloalkenyl" can be tetrahydropyridine, dihydropyran, dihydrofuran, pyrroline, dihydrobenzofuran, dihydrobenzothiophene, and indoline.

[0111] The term "aromatic" when applied to an entire substituent means a single ring or polycyclic ring system having 4n+2 electrons in a pi-conjugated system within the ring or ring system, and all atoms contributing to the pi-conjugated system lie in the same plane.

[0112] The term "aryl" refers to an aromatic hydrocarbon ring system. The ring system has 4n+2 electrons in a π-conjugated system within the ring, with all atoms contributing to the conjugated π system lying on the same plane. The ring system may be a single ring, or may be a bicyclic or tricyclic ring system. For example, "aryl" may be phenyl and naphthyl. The aryl system itself may be substituted with other groups.

[0113] The term "heteroaryl" refers to an aromatic hydrocarbon ring system having at least one heteroatom selected from O, N, and S in a single ring or fused ring system. The ring or ring system has 4n+2 electrons in a π-conjugated system, with all atoms contributing to the π-conjugated system lying in the same plane. The ring system may be a single ring, or a bicyclic or tricyclic ring system. For example, a "heteroaryl" can be imidazole, thien, furan, thianthrene, pyrrole, benzimidazole, pyrazole, pyrazine, pyridine, pyrimidine, and indole.

[0114] The term "alkaryl" refers to an aryl group as defined above, where C 1-4 This C refers to the one bound to an alkyl group. 1-4 The alkyl group provides the bond to the rest of the molecule.

[0115] The term "alkheteroaryl" refers to a heteroaryl group, as defined above, attached to a C1-4 alkyl, where the alkyl group provides the bond to the remainder of the molecule.

[0116] The term "halogen" as used herein includes reference to F, Cl, Br, and I. A halogen can be Cl. A halogen can be F.

[0117] A bond that terminates within a ring system and does not terminate at an atom of the ring system indicates that the bond may be attached to any atom in the ring system, as allowed by valences.

[0118] When a moiety is substituted, it can be substituted at any point on the moiety, within the limits of what is chemically possible and meets valence requirements. The moiety may be substituted as defined elsewhere herein, or with one or more substituents (e.g., 1, 2, 3, or 4 substituents), with one or two substituents optionally present on a group. When two or more substituents are present, the substituents may be the same or different. Substituents include -OH, -NH, amidoino, guanidino, hydroxyguanidino, formamidoino, isothioureido, ureido, mercapto, C(O)H, acyl, acyloxy, carboxy, sulfo, sulfamoyl, carbamoyl, cyano, azo, nitro, halo, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Haloalkyl, C 3-8 Cycloalkyl, C 2-6 Alkenyl, C 2-6 It may be selected from alkynyl, aryl, heteroaryl or alkaryl. When the substituted group is an alkyl group, the substituent may be =0.

[0119] Where chemically possible, cyclic substituents may be substituted on groups so as to form spirocyclic rings.

[0120] Substituents are present only at chemically possible positions, and one skilled in the art can determine (experimentally or theoretically) without undue effort which substitutions are chemically possible and which are not.

[0121] In embodiments in which the compounds of the present invention are single enantiomers, the compounds of the present invention may have an enantiomeric purity of at least about 90% enantiomeric excess (ee), at least about 95% enantiomeric excess (ee), at least about 98% enantiomeric excess (ee), at least about 99% enantiomeric excess (ee), or 100% enantiomeric excess (ee). In embodiments in which a mixture of enantiomers of the compounds of the present invention is present, the compounds of the present invention may be racemic or a mixture of other enantiomers. For example, the compounds of the present invention may have an enantiomeric purity of at least about 50% enantiomeric excess (ee), at least about 60% enantiomeric excess (ee), at least about 70% enantiomeric excess (ee), at least about 80% enantiomeric excess (ee), at least about 90% enantiomeric excess (ee), or at least about 95% enantiomeric excess (ee).

[0122] In this description, disclosure of a compound also encompasses its pharmaceutically acceptable salts, solvates, and stereoisomers. If a compound has a stereocenter, both the (R) and (S) stereoisomers are contemplated by the present invention, as are mixtures of stereoisomers or racemic mixtures. If a compound of the present invention has more than one stereocenter, any combination of the (R) and (S) stereoisomers is contemplated. Combinations of the (R) and (S) stereoisomers can result in mixtures of diastereoisomers or single diastereoisomers. The compounds of the present invention can exist as a single stereoisomer or as mixtures of stereoisomers (e.g., racemic mixtures and other enantiomeric mixtures) and diastereoisomers. When the mixture is a mixture of enantiomers, the enantiomeric excess can be any of those disclosed above. Even if a compound is a single stereoisomer, the compound may contain other diastereoisomers or enantiomers as impurities. Thus, a single stereoisomer does not necessarily have 100% enantiomeric excess (ee) or diastereomeric excess (de), but may have at least about 85% ee or de.

[0123] The present invention contemplates pharmaceutically acceptable salts of the compounds of Formula I. These may include acid addition or base salts of the compounds. These may be acid addition or base salts of the compounds. Additionally, the present invention contemplates solvates of the compounds. These may be hydrates or other solvated forms of the compounds.

[0124] Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include acetate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camsylate, citrate, edisylate, esylate, formate, fumarate, glucept, gluconate, glucuronate, hexafluorophosphate, hybenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfonate, naphthylate, 1,5-naphthalenedisulfonate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, saccharate, stearate, succinate, tartrate, tosylate, and trifluoroacetate.

[0125] Suitable base salts are formed from bases that form non-toxic salts. Examples include aluminum salt, arginine salt, benzathine salt, calcium salt, choline salt, diethylamine salt, diolamine salt, glycine salt, lysine salt, magnesium salt, meglumine salt, olamine salt, potassium salt, sodium salt, tromethamine salt, and zinc salt. Hemi-salts of acids and bases can also be formed, including, for example, hemisulfate salt and hemicalcium salt. For a discussion of suitable salts, see "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002).

[0126] Pharmaceutically acceptable salts of compounds of formula (I) may be prepared by one or more of three methods: (i) reacting a compound of formula (I) with a desired acid or base; (ii) removing an acid- or base-labile protecting group from a suitable precursor of a compound of formula (I), or ring-opening a suitable cyclic precursor (e.g., a lactone or lactam) with a desired acid or base; or (iii) Converting one salt of a compound of formula (I) into another salt by reaction with an appropriate acid or base or by using a suitable ion exchange column.

[0127] All three reactions are typically carried out in solution. The resulting salts may be precipitated and recovered by filtration or by evaporation of the solvent. The degree of ionization of the resulting salts may vary from completely ionized to mostly non-ionized.

[0128] The compound of the present invention can exist in both unsolvated and solvated form.The term "solvate" is used herein to describe a molecular complex that comprises the compound of the present invention and one or more stoichiometric amounts of pharmaceutically acceptable solvent molecules (e.g., ethanol).When the solvent is water, the term "hydrate" is used.

[0129] Hereinafter, references to compounds of any formula also include references to salts, solvates and complexes thereof and also to solvates and complexes of salts thereof.

[0130] The compounds of the present invention include compounds of the various formulas defined herein, including all polymorphs and crystalline forms thereof, prodrugs thereof, and isomers (including optical isomers, geometric isomers, and tautomers) defined below, and isotopically labeled compounds of the present invention.

[0131] Before purification, the compounds of the present invention may exist as a mixture of enantiomers, depending on the synthetic procedures used. Enantiomers can be separated by conventional techniques known in the art. Thus, the present invention encompasses individual enantiomers and mixtures thereof.

[0132] In some steps of the process for preparing compounds of formula (I), it may be necessary to protect potentially reactive functional groups that are not desired to react, followed by cleavage of the protecting groups. In such cases, any compatible protecting radical can be used. In particular, protection and deprotection methods such as those described by T.W.GREENE (Protective Groups in Organic Synthesis, A. Wiley-Interscience Publication, 1981) or P.J. Kocienski (Protecting groups, Georg Thieme Verlag, 1994) can be used. All of the above reactions, and the preparation of new starting materials used in the preceding methods, are conventional, and suitable reagents and reaction conditions for their implementation or preparation, as well as procedures for isolating the desired products, are well known to those skilled in the art by reference to the literature and the examples and preparations herein.

[0133] Additionally, the compounds of the present invention and intermediates for their preparation can be purified by various well-known methods, for example, crystallization or chromatography, etc.

[0178] The treatment methods or compounds for use in the treatment of cancer, lymphoma, leukemia, or immune disorders as defined above may be applied as monotherapy or in combination therapy with additional active agents.

[0134] Treatment methods or compounds used in the treatment of cancer, lymphoma, or leukemia may include, in addition to the compounds of the present invention, conventional surgery, radiation therapy, or chemotherapy. Such chemotherapy may include one or more of the following categories of anti-tumor agents: (i) Antiproliferative / antitumor drugs and combinations thereof, such as alkylating agents (e.g., cisplatin, oxaliplatin, carboplatin, cyclophosphamide, nitrogen mustard, bendamustine, melphalan, chlorambucil, busulfan, temozolomide, and nitrosoureas); antimetabolites (e.g., gemcitabine and antifolates, e.g., fluoropyrimidines such as 5-fluorouracil and tegafur, raltitrexed, methotrexate, pemetrexed, cytosine arabinoside, and hydroxyurea); antibiotics (e.g., adriamycin, bleomycin, doxorubicin, daun ... anthracyclines such as vincristine, epirubicin, idarubicin, mitomycin C, dactinomycin, and mithramycin; antimitotic agents (e.g., vinca alkaloids such as vincristine, vinblastine, vindesine, and vinorelbine, and taxoids such as paclitaxel and docetaxel, and polo kinase inhibitors); proteasome inhibitors (e.g., carfilzomib and bortezomib); interferon therapy; and topoisomerase inhibitors (e.g., epipodophyllotoxins such as etoposide and teniposide, amsacrine, topotecan, mitoxantrone, and camptothecin); (ii) cytostatics, such as antiestrogens (e.g., tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene, and iodoxifene), antiandrogens (e.g., bicalutamide, flutamide, nilutamide, and cyproterone acetate), LHRH antagonists or agonists (e.g., goserelin, leuprorelin, and buserelin), progestogens (e.g., megestrol acetate), aromatase inhibitors (e.g., anastrozole, letrozole, vorazole, and exemestane), and inhibitors of 5-alpha-reductase, such as finasteride; (iii) anti-invasive agents, such as dasatinib and bosutinib (SKI-606), and metalloproteinase inhibitors, urokinase-type plasminogen activator receptor function inhibitors, or antibodies against heparanase; (iv) Growth factor function inhibitors: For example, such inhibitors include growth factor antibodies and growth factor receptor antibodies, such as the anti-erbB2 antibody trastuzumab [Herceptin (trademark)], the anti-EGFR antibody panitumumab, the anti-erbB1 antibody cetuximab, tyrosine kinase inhibitors, such as inhibitors of the epidermal growth factor family (e.g., gefitinib, erlotinib, and 6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)-quinazolin-4-amine (CI EGFR family tyrosine kinase inhibitors such as EGFR-1033, erbB2 tyrosine kinase inhibitors such as lapatinib; inhibitors of the hepatocyte growth factor family; inhibitors of the insulin growth factor family; cell apoptosis-regulating protein modulators (e.g., Bcl-2 inhibitors); platelet-derived growth factor family inhibitors such as imatinib and / or nilotinib (AMN107); inhibitors of serine / threonine kinases (e.g., Ras / Raf signaling inhibitors such as farnesyltransferase, e.g., sorafenib, tipifarnib, and lonafarnib), inhibitors of cell signaling via MEK and / or AKT kinase, c-kit inhibitors, abl kinase inhibitors, PI3 kinase inhibitors, Plt3 kinase inhibitors, CSF-1 R kinase inhibitors, IGF receptor kinase inhibitors; Aurora kinase inhibitors, and cyclin-dependent kinase inhibitors such as CDK2 and / or CDK4 inhibitors; (v) antiangiogenic agents, such as those that inhibit the effects of vascular endothelial growth factor [e.g., the anti-vascular endothelial growth factor antibody bevacizumab (Avastin™); thalidomide; lenalidomide; and VEGF receptor tyrosine kinase inhibitors such as vandetanib, vatalanib, sunitinib, axitinib, and pazopanib; (vi) gene therapy approaches, including approaches to replace abnormal genes (e.g., abnormal p53 or abnormal BRCA1 or BRCA2); (vii) immunotherapeutic approaches, e.g., antibody therapies such as alemtuzumab, rituximab, ibritumomab tiuxetan (Zevalin®), and ofatumumab; interferons such as interferon a; interleukins such as IL-2 (aldesleukin); interleukin inhibitors (e.g., IRAK4 inhibitors); cancer vaccines, including preventative and therapeutic vaccines such as HPV vaccines, e.g., Gardasil, Cervarix, Oncophage, and Sipuleucel-T (Provenge); and Toll-like receptor modulators (e.g., TLR-7 or TLR-9 agonists); and (viii) cytotoxic agents, such as fludarabine (Fludara), cladribine, pentostatin (Nipent™); (ix) steroids such as corticosteroids, including glucocorticoids and mineralocorticoids, for example, aclometasone, aclomethasone dipropionate, aldosterone, amcinonide, beclomethasone, beclomethasone dipropionate, betamethasone, betamethasone dipropionate, betamethasone sodium phosphate, betamethasone valerate, budesonide, clobetasone, clobetasone butyrate, clobetasol propionate, cloprednol, cortisone, cortisone acetate, cortivazol, deoxycortone, desonide, desoximetasone, dexamethasone, dexamethasone sodium phosphate, dexamethasone isonicotinate, difluorocortolone, fluchlorolone, flumethasone, flunisolide, fluocinolone, fluocinolone acetonide, fluocinonide, fluocortin b til, fluorocortisone, fluorocortolone, fluocortolone caproate, fluocortolone pivalate, fluoromethalone, fluprednidene, fluprednidene acetate, flurandrenolone, fluticasone, fluticasone propionate, halcinonide, hydrocortisone, hydrocortisone acetate, hydrocortisone butyrate, hydrocortisone aceponate, hydrocortisone butyrate propionate, hydrocortisone valerate, icometasone, icometasone embutate, meprednisone, methylprednisolone, mometasone, paramethasone, mometasone furoate hydrate, prednicarbate, prednisolone, prednisone, tixocortol, tixocortol pivalate, triamcinolone, triamcinolone acetonide, triamcinolone alcohol, and pharmaceutically acceptable derivatives thereof. Combinations of steroids may be used, for example, a combination of two or more of the steroids described in this paragraph; (x) Targeted therapies, such as PI3Kd inhibitors, such as idelalisib and perifosine.

[0135] Such combination treatment may be achieved by the simultaneous, sequential, or separate administration of the individual components of the treatment. Such combination products utilize the compounds of this invention within the therapeutically effective dosage range described above and the other pharmaceutically active agent(s) within their approved dosage ranges.

[0136] According to a further aspect of the present invention there is provided a pharmaceutical product comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof as defined above, and an additional active agent, which may be an anti-tumour agent as defined above for the combined treatment of cancer.

[0137] According to a further aspect of the present invention there is provided a method of treating a condition treatable by a cytotoxic agent comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, simultaneously, sequentially or separately with an additional anti-tumor agent as defined above.

[0138] According to a further aspect of the present invention there is provided a compound of formula (I), or a pharmaceutically acceptable salt thereof, for use simultaneously, sequentially or separately with an additional anti-tumour agent as defined above in the treatment of a condition treatable by a cytotoxic agent.

[0139] According to another aspect of the present invention, there is provided the use of a compound of formula (I) in combination with an antitumor agent as described above. The compound of formula (I) may be used simultaneously, sequentially, or separately with the additional antitumor agent. The use may be in a single combination product containing the compound of formula (I) and the antitumor agent.

[0140] According to a further aspect, there is provided a method of providing a combination product, the method comprising simultaneously, sequentially or separately providing a compound of formula (I) and an anti-tumor agent as defined above. The method may comprise combining the compound of formula (I) and the anti-tumor agent in a single dosage form. Alternatively, the method may comprise providing the anti-tumor agent in a separate dosage form.

[0141] According to a further aspect, there is provided a method of providing a combination product, the method comprising simultaneously, sequentially or separately providing a compound of formula (I) and an anti-tumor agent as defined above. The method may comprise combining the compound of formula (I) and the anti-tumor agent in a single dosage form. Alternatively, the method may comprise providing the anti-tumor agent in a separate dosage form.

[0142] [Example]

[0143] The compounds of the present invention were prepared by the following method.

[0144] General Method A:

[0145] General method for the synthesis of 5-alkyl / arylamino-(1-(chloromethyl)-1,2-dihydro-3H-benzo[e]indol-3-yl)(5-methoxy-1H-indol-2-yl)methanones

[0146] 5-Triflyloxy-(1-(chloromethyl)-1,2-dihydro-3H-benzo[e]indol-3-yl)(5-methoxy-1H-indol-2-yl)methanone (27 mg, 0.05 mmol), potassium phosphate (12 mg, 0.1 mmol), Xantphos (12 mg, 0.02 mmol), and the appropriate alkyl or arylamino derivative (0.15 mmol) were mixed in dry toluene (1.5 mL) and heated under nitrogen in an oven-dried pressure tube. Pd(OAc) (2.2 mg, 0.01 mmol) was added. The reaction mixture was stirred and refluxed in an oil bath at 110-125 °C for 1-4 h. The crude reaction mixture was directly applied to a silica gel column to obtain the purified product.

[0147] Example 1 - DP58 [ka]

[0148] HPLC-MS: m / z(%)=496[M+H], 498[M+2 + H]

[0149] LC / MS analysis was performed using a Waters analytical system containing the following modules: a Waters Alliance 2695 separation module, a Waters 996 PDA detector, and a Waters Micromass ZQ mass detector.

[0150] Column: Hichrom RPB-150 AM, 2.1x150mm, 3.5μm; Mobile phase A: 90% water, 10% methanol + 0.1% formic acid; Mobile phase B: 90% methanol, 10% water + 0.1% formic acid; Flow rate 0.25ml / min. [Table 1]

[0151] 1H NMR (CDCl3) δ: 9.47 (s, 1H), 7.88 (s, 1H), 7.81 (d, J = 8 Hz, 1H), 7.72 (dd, J = 8 / 0.8 Hz 1H), 7.52 (m, 1H), 7.45 (m, 2H), 7,36 (m, 5H), 7.13 (d, J = 2.4 Hz, 1H), 7.04 (dd, J = 2.8 / 0.8 Hz 1H), 7.00 (dd, J = 6.4 / 2.4 Hz 1H), 4.79 (m, 2H), 4.64 (m, 1H), 4.52 (m, 2H), 4.09 (m, 1H), 3.96 (m, 1H), 3.87 (s, 3H, OMe), 3.44 (t, J = 10.8 Hz 1H).

[0152] Example 2 - DP-59 [ka]

[0153] HPLC-MS: m / z(%)=512[M+H], 514[M+2 + H]

[0154] Example 3 - DP-63 [ka]

[0155] HPLC-MS: m / z(%)=462[M+H], 464[M+2 + H]

[0156] Example 4 - DP-65 [ka]

[0157] HPLC-MS: m / z(%)=534[M+H], 536[M+2 + H]

[0158] Example 5 - DP-70 [ka]

[0159] HPLC-MS: m / z(%)=550[M+H], 552[M+2 + H]

[0160] Common Method B:

[0161] General method for the synthesis of 5-allyl-(1-(chloromethyl)-1,2-dihydro-3H-benzo[e]indol-3-yl)(5-methoxy-1H-indol-2-yl)methanone

[0162] To a mixture of 5-triflyloxy-(1-(chloromethyl)-1,2-dihydro-3H-benzo[e]indol-3-yl)(5-methoxy-1H-indol-2-yl)methanone (27 mg, 0.05 mmol), potassium fluoride (10.5 mg, 0.18 mmol), BINAP (6 mg, 0.01 mmol), and the appropriate substituted phenylboronic acid (0.15 mmol) in dry toluene (1.5 mL) was added Pd(OAc) (1.1 mg, 0.005 mmol) in an oven-dried pressure tube under a nitrogen atmosphere. The reaction mixture was stirred and refluxed in an oil bath at 100 °C for 1 h. The crude reaction mixture was applied directly to a silica gel column to give the purified product.

[0163] Example 6 - DP-36 [ka]

[0164] HPLC-MS: m / z(%)=483[M+H], 485[M+2 + H]

[0165] Example 7 - DP-44 [ka]

[0166] HPLC-MS: m / z(%)=497[M+H], 499[M+2 + H]

[0167] Example 8 - DP-45 [ka]

[0168] HPLC-MS: m / z(%)=481[M+H], 483[M+2 + H]

[0169] Example 9 – DP-47 [ka]

[0170] HPLC-MS: m / z(%)=483[M+H], 485[M+2 + H]

[0171] Example 10 - DP-48 [ka]

[0172] HPLC-MS: m / z(%)=482[M+H], 484[M+2 + H]

[0173] Common synthetic C:

[0174] General method for ester synthesis using alkynoic acids

[0175] To a mixture of (5-(benzyloxy)-1-(chloromethyl)-1,2-dihydro-3H-benzo[e]indol-3-yl)(5-hydroxy-1H-indol-2-yl)methanone (48 mg, 0.1 mmol), triethylamine (42 μL, 0.3 mmol), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (38 mg, 0.2 mmol) in dry dichloromethane (3 mL) was added the appropriate alkynol acid (0.15 mmol) in an oven-dried flask. The reaction mixture was stirred at room temperature for 15 minutes. The crude reaction mixture was applied directly to a silica gel column to give the purified product.

[0176] Example 11 - DP-42 [ka]

[0177] HPLC-MS: m / z(%)=563[M+H], 565[M+2 + H]

[0178] 1H NMR (CDCl3) δ: 9.82 (s, 1H), 8.36 (dd, J = 8 / 0.8 Hz, 1H), 8.20 (s, 1H), 7.70 (d, J = 8 Hz, 1H), 7.58-7.51 (m, 3H), 7.46-7.40 (m, 5H), 7.37-7.33 (m, 1H), 7.07 (d, J = 1.4 Hz 1H), 7.03 (dd, J = 8.8 / 2.4 Hz, 1H), 5.28 (m, 2H), 4.79 (dd, J = 8 / 0.8 Hz, 1H), 4.64 (m, 1H), 4.11 (m, 1H), 3.97 (dd, J = 8.4 / 2.4 Hz 1H), 3.47 (t, J = 10.8 Hz, 1H), 2.85 (m, 2H), 2.68 (m, 2H), 2.07 (t, J = 2.8 Hz, 1H).

[0179] Example 12 - DP-51

change

[0180] HPLC-MS:m / z(%)=577[M+H], 579[M+2 + H]

[0181] Example 13 - DP-77

change

[0182] HPLC-MS:m / z(%)=497[M+H], 499[M+2 + H]

[0183] Example 14 - DP-78

change

[0184] HPLC-MS:m / z(%)=407[M+H], 409[M+2 + H]

[0185] 1H NMR (CDCl3) δ: 11.64 (s, 1H), 9.87 (s, 1H), 8.17 (d, J = 0.8 Hz, 1H), 7.78 (m, 2H), 7.40 (d, J = 8.8 Hz, 1H), 7.16-7.10 (m, 3H), 7.02 (dd, J = 8.8 / 1.2 Hz, 1H), 6.92 (dd, J = 8.8 / 2.0 Hz, 1H), 4.80 (t, J = 10 Hz, 1H), 4.58 (d, J = 10.4 Hz 1H), 4.21 (m, 1H), 4.04 (dd, J = 10 / 2.4 Hz, 1H), 3.88 (dd, J = 10.8 / 7.6 Hz, 1H), 3.78 (s, 3H).

[0186] Example 15 - DP-31

[0187] The synthesis of Examples 15 and 16 is based on known methods disclosed in Boger et al., J. Org. Chem., 1999, 64(14), 5241-5244, and Sheldrake et al., J. Med. Chem., 2013, 56(15), 6273-7. [ka]

[0188] HPLC-MS: m / z(%)=407[M+H], 409[M+2 + H]

[0189] Example 16 - DP-40 [ka]

[0190] HPLC-MS: m / z(%)=483[M+H], 485[M+2 + H]

[0191] Example 17 - tert-Butyl 1-(chloromethyl)-5-phenyl-1H-benzo[e]indole-3(2H)-carboxylate (ICT-11081) [ka]

[0192] Synthesized by Method B

[0193] 1 H NMR (400 MHz, CDCl3) δ 8.16 (bs, 1H), 7.85 (d, 1H, J = 8.4 Hz), 7.76 (d, 1H, J = 8.0 Hz), 7.51 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.0 Hz), 7.47-7.39 (m, 5H) 7.26 (ddd, 1H, J= 6.8 Hz, J = 1.6 Hz, J = 8.4 Hz), 4.31 (d, 1H, J = 11.4 Hz), 4.15 (dd, 1H, J = 8.8 Hz, J = 11.4 Hz), 4.09 (m, 1H), 4.00 (dd, 1H, J = 2.4 H, J = 11.2 Hz), 3.52 (dd, 1H, J = 10.4 Hz, J = 11.2 Hz), 1.57 (s, 9H); 13 C NMR (100 MHz, CDCl3) δ 152.61, 140.67, 130.00 (2C), 128.19 (2C), 127.49, 127.38 (2C), 126.96 (2C), 123.94, 122.31, 116.90, 67.09, 53.43, 52.73, 46.32, 28.44 (3C); HRMS (ESI+) is C 24 H 25 ClNO2394.15738 [M+H] + The calculated value was 394.08164 cacld.

[0194] Example 18 - tert-Butyl 1-(chloromethyl)-5-(4-hydroxyphenyl)-1H-benzo[e]indole-3(2H)-carboxylate (ICT-11071) [ka]

[0195] Synthesized by Method B

[0196] 1 H NMR (400 MHz, CDCl3) δ 8.17 (bs, 1H), 7.87 (d, 1H, J = 8.4 Hz), 7.76 (d, 1H, J = 8.0 Hz), 7.50 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.0 Hz), 7.33 (d, 2H, J = 8.6 Hz), 7.30 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.4 Hz), 6.93 (d, 2H, J = 8.6 Hz), 4.98 (bs, 1H), 4.32 (d, 1H, J = 11.6 Hz), 4.16 (dd, 1H, J = 11.6 Hz, J = 8.4 Hz), 4.07 (m, 1H), 4.00 (dd, 1H, J = 2.8 Hz, J = 11.6 Hz), 3.50 (dd, 1H, J = 10.4 Hz, J = 11.6 Hz), 1.58 (s, 9H); 13 C NMR (100 MHz, CDCl3) δ 154.45, 152.64, 130.97 (2C), 127.42, 126.71, 123.93, 116.59, 115.24 (2C), 55.96, 46.33, 35.48, 20.47 (3C); HRMS (ESI+) is C 24 H 25 ClNO3410.15230 [M+H] + The result was 410.15188 calcd.

[0197] Example 19 - tert-Butyl 1-(chloromethyl)-5-(4-methoxyphenyl)-1H-benzo[e]indole-3(2H)-carboxylate (ICT-11082) [ka]

[0198] Synthesized by Method B

[0199] 1 H NMR (400 MHz, CDCl3) δ 7.89 (d, 1H, J = 8.5 HZ), 7.76 (d, 1H, J = 8.3 Hz), 7.48 (dd, 1H, J= 6.8 Hz, J = 8.3 Hz), 7.40 (d, 2H, J = 8.8 Hz), 7.30 (dd, 1H, J= 6.8 Hz, J = 8.5 Hz), 7.01 (d, 2H, J = 8.8 Hz), ), 6.96 (dt, J=8.78, 1H), 4.32 (d, 1H, J = 11.0 Hz), 4.16 (dd, 1H, J = 8.8 Hz, J = 11.0 Hz), 4.07 (m, 1H), 4.00 (dd, 1H, J = 2.6 Hz, J = 10.9 Hz), 3.89 (s, 3H), 3.49 (dd, 1H, J = 6.3 Hz, J = 10.9 Hz), 1.58 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 131.27 (s, 2C), 127.88 (s), 127.69 (s), 123.98 (s), 122.43 (s), 114.31 (s), 113.80 (s, 2C), 55.50 (s), 52.88 (s), 46.46 (s), 42.22, 28.60 (s, 3C);

[0200] Example 20 - (1-(chloromethyl)-5-phenyl-1H-benzo[e]indol-3(2H)-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11043) [Chemical]

[0201] Synthesized by Method B

[0202] 1 1H NMR (400 MHz, CDCl3) δ 9.52 (s, 1H, NH), 8.54 (s, 1H), 7.89 (d, 1H, J = 8.4 Hz), 7.86 (d, 1H, J = 8.0 Hz), 7.56 (ddd, 1H J = 1.2 Hz, J = 6.8 Hz, J = 8.0 Hz), 7.51 - 7.42 (m, 5H), 7.37 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.4 Hz), 7.36 (d, 1H, J = 9.2 Hz), 7.13 (d, 1H, J = 2.4 Hz), 7.07 (dd, 1H, J = 0.8 Hz, J = 2.0 Hz), 6.98 (dd, 1H, J = 2.4 Hz, J = 9.2 Hz), 4.89 (dd, 1H, J = 2.0 Hz, J = 10.8 Hz), 4.73 (dd, 1H, J = 8.4 Hz, J = 10.8 Hz), 4.27 (m, 1H), 4.05 (dd, 1H, J = 2.8 Hz, J = 11.2 Hz), 3.88 (s, 3H), 3.56 (dd, 1H, J = 10.5 Hz, J = 11.2 Hz); 13 13C NMR (100 MHz, CDCl3) δ 160.67, 154.74, 142.59, 141.17, 140.43, 131.35, 130.49, 130.12 (2C), 129.81, 129.73, 128.30 (2C), 127.64, 127.54, 127.18, 12 4.89, 123.94, 122.68, 118.79, 116.84, 115.31, 112.75, 106.07, 10 2.47, 55.75, 55.05, 45.87, 43.69; HRMS (ESI+) is C 29 H 24ClN2O2467.15263 [M+H] + The result was 467.15180 calcd.

[0203] Example 21 - (1-(chloromethyl)-5-(2-hydroxyphenyl)-1H-benzo[e]indol-3(2H)-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11044) [ka]

[0204] Synthesized by Method B

[0205] 1 H NMR (400 MHz, CDCl3) δ 9.29 (s, 1H, NH), 8.59 (s, 1H), 7.80 (d, 1H, J = 8.4 Hz), 7.62 (dd, 1H, J = 4.0 Hz, J = 8.4 Hz), 7.52 (m, 1H), 7.36-7.28 (m, 3H), 7.28 (dd, 1H, J = 1.2 Hz, J = 7.2 Hz), 7.16 (dd, 1H, J = 1.6 Hz, J = 7.6 Hz), 7.07 (d, 1H, J = 2.4 Hz), 7.01-6.93 (m, 4H), 4.84 (dd, J = 2.0 Hz, J = 10.8 Hz), 4.69 (dd, 1H, J = 6.4 Hz, J = 10.8 Hz), 4.29 (m, 1H), 4.07 (dd, 1H, J = 3.2 Hz, J =11.6 Hz), 3.70 (s, 3H), 3.59 (dd, 1H, J = 10.8 Hz, J = 11.6 Hz); HRMS (ESI+) is C 29 H 24 ClO3N2483.14755 [M+H] + The result was 483.14694 calcd.

[0206] Example 22 - (1-(chloromethyl)-5-(4-methoxyphenyl)-1H-benzo[e]indol-3(2H)-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11038) [ka]

[0207] Synthesized by Method B

[0208] 1 H NMR (400 MHz, CDCl3) δ 9.29 (s, 1H, NH), 8.50 (s, 1H), 7.94 (d, 1H, J = 8.2 Hz), 7.84 (d, 1H, J = 8.2 Hz), 7.56 (m, 1H), 7.43 (m, 2H), 7.37 (m, 2H), 7.14 (d, 1H, J = 2.2 Hz), 7.07 (dd, 1H, J = 2.2 Hz, J = 0.4 Hz), 7.02 (m, 3H), 4.89 (dd, 1H, J = 10.8 Hz, J = 1.6 Hz), 4.74 (m, 1H), 4.26 (m, 1H) 4.06 (dd, 1H, J = 11.3 Hz, J = 3.1 Hz), 3.9 (s, 3H), 3.88 (s, 3H), 3.554 (dd, 1H, J = 10.8 Hz, J = 11.3 Hz); 13 C NMR (100 MHz, CDCl3) δ 160.52, 159.17, 154.77, 143.94, 142.31, 141.21, 132.81, 131.22 (2C), 130.55, 129.92, 129.83, 128.37, 127.69, 127.12, 124.78, 123.57, 122.66, 118.75, 116.86, 113.74, 112.71, 106.03, 102.53, 55.75, 55.39, 54.97, 45.89, 43.69; HRMS (ESI + ) is C 30 H 26ClN2O3497.16320 [M+H] + The result was 497.16526 calcd.

[0209] Example 23 - (1-(chloromethyl)-5-(4-(methoxymethyl)phenyl)-1H-benzo[e]indol-3(2H)-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11039) [ka]

[0210] Synthesized by Method B

[0211] 1 H NMR (400 MHz, CDCl3) δ 9.31 (s, 1H, NH), 8.52 (s, 1H), 7.91 (d, 1H, J = 8.8 Hz), 7.85 (d, 1H, J = 8.0 Hz), 7.56 (m, 1H), 7.4815 (m, 4H), 7.36 (d, 2H, J= 8.8 Hz), 7.14 (d, 1H, J = 2.4 Hz), 7.07 (m, 1H), 7.01 (dd, 1H, J= 8.8 Hz, J = 2.4 Hz), 4.89 (dd, 1H, J = 10.8 Hz, J = 2.0 Hz), 4.74 (m, 1H), 4.56 (s, 2H), 4.27 (m, 1H), 4.06 (dd, 1H, J = 11.2 Hz, J = 2.8 Hz), 3.88 (s, 3H), 3.56 dd, 1H, J = 10.8 Hz, J = 11.2 Hz), 3.47 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 160.88, 155.11, 142.67, 141.53, 140.17, 137.88, 131.57, 130.84, 130.53 (2C), 130.14, 130.07, 128.70, 128.04 (2C), 127.97, 127.52, 125.51, 124.23, 123.00, 119.11, 117.21, 113.05, 106.38, 102.86, 74.89, 58.63, 56.08, 55.30, 46.22, 44.03; HRMS (ESI) is C 31 H 28 ClN2O3511.17885 [M+H] + The result was 511.17814 calcd.

[0212] Example 24 - (1-(chloromethyl)-5-(pyridin-4-yl)-1H-benzo[e]indol-3(2H)-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11040) [ka]

[0213] Synthesized by Method B

[0214] 1H NMR (400 MHz, CDCl3) δ 9.36 (s, 1H, NH), 8.75 (dd, 2H, J = 4.4 Hz, J = 1.6 Hz), 8.54 (s, 1H), 7.86 (d, 2H, J = 8.0 Hz), 7.61 (dd, 1H, J = 7.8 Hz, J = 1.2 Hz), 7.45 (d, 2H, J = 6 Hz), 7.42 (m, 1H), 7.36 (d, 1H, J = 8.8 Hz), 7.14 (d, 1H, J = 2.4 Hz), 7.08 (dd, 1H, J= 2 Hz, J = 0.8 Hz), 7.01 (dd, 1H, J = 8.8 Hz, J = 2.4 Hz), 4.91 (dd, 1H, J = 10.8 Hz, J = 2.0 Hz), 4.75 (m, 1H), 4.29 (m, 1H), 4.06 (m, 1H), 3.87 (s, 3H), 3.58 (dd, 1H, J = 10.4 Hz, J= 10.8 Hz); 13 C NMR (100 MHz, CDCl3) δ 161.0, 155.1, 150.2, 148.8, 141.5, 139.7, 131.6, 130.5, 130.2, 129.1, 128.6, 127.9, 127.1, 125.8, 125.4, 123.0, 119.1, 117.3, 113.0, 106.5, 102.8, 56.0, 55.2, 45.1, 43.9; HRMS (ES+I) is C 28 H 23 ClN3O2468.14778 [M+H] + The result was 468.14703 calcd.

[0215] Example 25 - (1-(chloromethyl)-5-(pyridin-3-yl)-1H-benzo[e]indol-3(2H)-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11041) [ka]

[0216] Synthesized by Method B

[0217] 1 H NMR (400 MHz, CDCl3) δ 9.51 (s, 1H, NH), 8.78 (d, 1H, J = 1.6 Hz), 8.72 (dd, 1H, J = 1.6 Hz, J = 4.9 Hz), 8.55 (s, 1H), 7.88 (d, 1H, J = 8.6 Hz), 7.85 (m, 1H), 7.81 (d, 1H, J = 8.4 Hz), 7.6 (ddd, 1H, J = 1.1 Hz, J = 6.8 Hz, J = 8.4 Hz), 7.46 (ddd, 1H, J = 2.1 Hz, J = 4.5 Hz, J = 6.3 Hz), 7.43 (dd, 1H, J = 1.3 Hz, J = 7.0 Hz), 7.41 (dd, 1H, J = 1.3 Hz, J = 6.9 Hz), 7.36 (d, 1H, J = 8.9 Hz), 7.14 (d, 1H, J = 2.4 Hz), 7.08 (d, 1H, J = 2.0 Hz), 7.00 (dd, 1H, J = 2.4 Hz, J = 8.9 Hz), 4.91 (dd, 1H, J = 1.9 Hz, J = 10.8 Hz), 4.76 (dd, 1H, J = 8.7 Hz, J = 10.8 Hz), 4.29 (m, 1H), 4.06 (dd, 1H, J = 2.7 Hz, J = 11.3 Hz), 3.88 (s, 3H) 3.58 (dd, 1H, J = 10.5 Hz, J = 11.3 Hz); 13 C NMR (100 MHz, CDCl3) δ 160.75, 154.80, 150.72, 148.92, 137.98, 131.40, 128.30, 127.52, 126.88, 125.43, 124.89, 123.37, 122.90, 119.32, 117.01, 112 / 79, 106.23, 102.84, 55.99, 55.29, 46.11, 44.04; HRMS (ESI+) is C 28 H 23468.14988 calcd was calculated for ClO2N3468.14788 [M+H]+.

[0218] Example 26 - (1-(chloromethyl)-5-(4-(dimethylamino)phenyl)-1H-benzo[e]indol-3(2H)-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11042) [ka]

[0219] Synthesized by Method B

[0220] 1 H NMR (400 MHz, CDCl3) δ 9.34 (s, 1H, NH), 8.50 (s, 1H), 8.04 (d, 1H, J = 8.0 Hz), 7.85 (d, 1H, J = 8.4 Hz), 7.54 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.4 Hz), 7.42 (d, 2H, J = 8.8 Hz), 7.39 (ddd, 1H, J= 1.2 Hz, J = 6.8 Hz, J = 8.0 Hz), 7.37 (d, 1H, J = 8.9 Hz), 7.14 (d, 1H, J = 2.4 Hz), 7.06 (d, 1H, J = 1.9 Hz), 7.00 (dd, 1H, J = 2.4 Hz, J = 8.9 Hz), 6.85 (d, 2H, J = 8.8 Hz), 4.88 (dd, 1H, J = 1.7 Hz, J = 10.8 Hz), 4.72 (dd, J = 8.7 Hz, J = 10.8 Hz), 4.24 (m, 1H), 4.06 (dd, 1H, J = 3.1 Hz, J = 11.1 Hz), 3.88 (s, 3H), 3.54 (dd, 1H, J = 10.4 Hz, J = 11.1 Hz), 2.04 (s, 6H); 13C NMR (100 MHz, CDCl3) δ 160.50, 154.74, 149.99, 142.98, 141.28, 131.22, 130.94 (2C), 130.66, 130.02, 129.91, 128.40, 127.97, 127.99, 124.55, 123.05, 122.59, 118.52, 116.77, 112 / 71, 112.20 92C), 105.95, 102.51, 56.04, 55.13, 46.91, 43.83, 41.36, 40.61; HRMS (ESI+) is C 31 H 29 ClO2N3510.19483 [M+H] + The result was 510.19404 calcd.

[0221] Example 27 - (5-(6-aminopyridin-3-yl)-1-(chloromethyl)-1H-benzo[e]indol-3(2H)-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11083) [ka]

[0222] Synthesized by Method B

[0223] 1H NMR (400 MHz, CDCl3) δ 9.29 (s, 1H, NH), 8.48 (s, 1H), 8.24 (s, 1H), 7.90 (d, 1H, J= 8.4 Hz), 7.84 (d, 1H, J = 8.4 Hz), 7.62 (dd, 1H, J = 8.6 Hz, J= 2.4 Hz), 7.57 (m, 1H), 7.55 (m, 1H), 7.42 (d, 1H, J = 1.2 Hz), 7.39 (m, 1H), 7.14 (d, 1H, J = 2.4 Hz), 7.07 (m, 1H), 7.03 (m, 1H), 6.67 (d, 1H, J = 8.4 Hz), 4.89 (m, 1H), 4.73 (m, 1H), 4.26 (m, 1H), 4.05 (dd, 1H, J = 11.2 Hz, J = 3.2 Hz), 3.88 (s, 3H), 3.55 (dd, 1H, J = 10.8 Hz, J = 11.2 Hz);

[0224] Example 28 - (1-(chloromethyl)-5-(3-fluoro-4-hydroxyphenyl)-1,2-dihydro-3H-benzo[e]indol-3-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11075) [ka]

[0225] Synthesized by Method B

[0226] 1H NMR (400 MHz, acetone-d6) δ 10.79 (s, 1H, NH), 8.88 (s, 1H), 8.51 (s, 1H), 8.05 (d, 1H, J = 8.0 Hz), 7.9 (d, 1H, J = 8.4 Hz), 7.59 (ddd, 1H, J= 1.2 Hz, J = 6.8 Hz, J = 8.0 Hz), 7.47 (d, 1H, J = 8.8 Hz), 7.42 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.4 Hz), 7.26 (m, 1H), 7.18 (m, 4H), 6.94 (dd, 1H, J = 2.4 Hz, J = 8.8 Hz), 4.86 (m, 2H), 4.45 (m, 1H), 4.15 (dd, 1H, J = 3.4 Hz, J = 11.2 Hz), 3.94 (dd, 1H, J = 8.4 Hz, J = 11.2 Hz), 3.83 (s, 3H, OMe); 13 C NMR (100 MHz, acetone-d6) δ 161.3, 155.5, 142.7, 141.2, 133.5 (2C), 132.7, 132.0, 131.0, 130.2, 129.2, 127.9, 127.7, 127.1 (2C), 125.6, 125.3, 124.1, 119.6, 118.6, 118.4, 118.2, 116.9, 113.9, 106.5, 103.0, 55.7, 47.6, 43.5;

[0227] Example 29 - 4-(1-(chloromethyl)-3-(5-methoxy-1H-indole-2-carbonyl)-2,3-dihydro-1H-cyclopenta[a]naphthalen-5-yl)phenyl acetate (ICT-11072) [ka]

[0228] Synthesized by Method B

[0229] 1H NMR (400 MHz, acetone-d6) δ 10.79 (s, 1H, NH), 8.53 (s, 1H), 8.06 (d, 1H, J= 8.4 Hz), 7.84 (d, 1H, J = 8.4 Hz), 7.59 (dd, 1H, J = 7.6 Hz, J = 8.4 Hz), 7.54 (d, 2H, J = 8.4 Hz), 7.47 (d, 1H, J= 8.8 Hz), 7.42 (dd, 1H, J = 7.6 Hz, J = 8.4 Hz), 7.30 (d, 2H, J= 8.4 Hz), 7.19 (s, 2H), 6.94 (dd, 1H, J = 2.6 Hz, J = 8.8 Hz), 4.98 (m, 2H), 4.45 (m, 1H), 4.16 (dd, 1H, J = 3.2 Hz, J = 11.2 Hz), 3.95 (dd, 1H, J = 8.4 Hz, J = 11.2 Hz), 3.83 (s, 3H), 2.31 (s, 3H); 13 C NMR (100 MHz, acetone-d6) δ 169.7, 161.3, 155.5, 151.5, 142.7, 141.6, 139.0, 132.7, 132.0, 131.7, 131.0, 130.1, 129.2, 127.9, 127.6, 125.7, 125.5, 124.2, 123.3, 122.7, 119.6, 116.9, 113.9, 106.5, 103.0, 55.7, 47.6, 43.5, 21.0;

[0230] Example 30 - (1-(chloromethyl)-5-(4-hydroxy-3-nitrophenyl)-2,3-dihydro-1H-cyclopenta[a]naphthalen-3-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11069) [ka]

[0231] Synthesized by Method B

[0232] 1 H NMR (400 MHz, THF-d8) δ 10.93 (s, 1H, NH), 10.56 (s, 1H), 8.58 (s, 1H), 8.21 (d, 1H, J= 2.4 Hz), 8.0 (d, 1H, J = 8.4 Hz), 7.79 (d, 1H, J = 8.4 Hz), 7.76 (dd, 1H, J = 2.2 Hz, J = 8.6 Hz), 7.56 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.4 Hz), 7.37 (ddd, 1H, J = 1.6 Hz, J = 6.8 Hz, J = 8.4 Hz), 7.34 (d, 1H, J = 9.2 Hz), 7.3 (d, 1H, J = 8.8 Hz), 7.11 (m, 2H), 6.9 (dd, 1H, J = 2.4 Hz, J= 9.2 Hz), 4.85 (m, 2H), 4.36 (m, 1H), 4.09 (dd, 1H, J = 3.2 Hz, J= 11.2 Hz), 3.8 (s, 3H), 3.74 (dd, 1H, J = 9.4 Hz, J = 11.2 Hz); 13 C NMR (100 MHz, THF-d8) δ 161.4, 155.7, 142.9, 139.9, 139.4, 135.2, 133.8, 133.0, 132.0, 131.2, 130.1, 129.3, 127.9, 127.3, 126.8, 125.8, 125.7, 124.1, 121.0, 120.0, 116.9, 113.5, 106.3, 102.8, 55.9, 55.6, 47.1, 44.2;

[0233] Example 31 - (1-(chloromethyl)-5-(4-hydroxy-2-methylphenyl)-2,3-dihydro-1H-cyclopenta[a]naphthalen-3-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11070) [ka]

[0234] Synthesized by Method B

[0235] 1 H NMR (400 MHz, acetone-d6) δ 10.77 (s, 1H, NH), 8.42 (bs, 1H), 8.41 (d, 1H,J = 1.6 Hz), 8.04 (dd, 1H, J = 0.8 Hz, J = 8.4 Hz), 7.56 (m, 1H), 7.46-7.5 (m, 2H), 7.36 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J= 8.4 Hz), 7.19 (m, 2H), 7.07 (m, 1H), 6.94 (dd, 1H, J = 2.4 Hz, J= 9.2 Hz), 6.87 (dd, 1H, J = 2.6 Hz, J = 4.8 Hz), 6.82 (td, 1H, J= 2.6 Hz, J = 8.6 Hz, H6''), 4.91 (m, 2H), 4.47 (m, 1H), 4.18 (m, 1H), 3.95 (ddd, 1H, J = 3.6 Hz, J = 8.4 Hz, J = 11.2 Hz); 3.83 (s, 3H), 1.99 (s, 3H, Me); 13 C NMR (100 MHz, acetone-d6) δ 161.2, 155.5, 146.4, 146.0, 142.1, 141.1, 140.5, 136.1, 132.7, 132.0, 131.6, 130.8, 129.2, 127.9, 127.7, 125.3, 124.9, 124.1, 119.7, 117.5, 116.8, 113.9, 113.5, 106.4, 103.0, 55.7, 47.6, 43.7, 20.3;

[0236] Example 32 - (1-(chloromethyl)-5-(4-hydroxy-3-methylphenyl)-2,3-dihydro-1H-cyclopenta[a]naphthalen-3-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11079) [ka]

[0237] Synthesized by Method B

[0238] 1 H NMR (400 MHz, THF-d8) δ 10.86 (s, 1H, NH), 8.47 (s, 1H), 8.3 (s, 1H), 7.91 (m, 2H), 7.49 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.4 Hz), 7.34 (d, 1H, J = 8.8 Hz), 7.29 (ddd, 1H, J = 1.2 Hz, J= 6.8 Hz, J = 8.4 Hz), 7.18 (d, 1H, J = 2.0 Hz), 7.09 (m, 2H), 7.07 (dd, 1H, J = 0.8 Hz, J = 2.0 Hz), 6.89 (dd, 1H, J= 2.4 Hz, J = 8.8 Hz), 6.81 (d, 1H, J = 8.4 Hz), 4.81 (m, 2H), 4.29 (m, 1H), 4.07 (dd, 1H, J = 2.8 Hz, J = 11.2 Hz), 3.79 (s, 3H), 3.68 (dd, 1H, J = 9.8 Hz, J = 11.2 Hz), 2.25 (s, 3H, Me); 13 C NMR (100 MHz, THF-d8) δ 161.3, 156.3, 155.6, 143.3, 142.8, 133.3, 133.0, 132.7, 132.3, 131.1, 130.7, 129.3, 129.1, 128.4, 127.4, 125.1, 124.8, 124.2, 123.7, 119.6, 116.7, 115.0, 113.4, 106.1, 102.9, 55.9, 55.6, 47.0, 44.3, 16.3;

[0239] Example 33 - (1-(chloromethyl)-5-(4-nitrophenyl)-2,3-dihydro-1H-cyclopenta[a]naphthalen-3-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11078) [ka]

[0240] Synthesized by Method B

[0241] 1 H NMR (400 MHz, THF-d8) δ 10.89 (s, 1H, NH), 8.61 (s, 1H), 8.38 (d, 2H, J= 8.8 Hz), 8.01 (d, 1H, J = 8.2 Hz), 7.75 (m, 3H), 7.57 (ddd, 1H, J= 1.2 Hz, J = 6.8 Hz, J = 8.2 Hz), 7.38 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.4 Hz), 7.34 (d, 1H, J = 8.8 Hz), 7.11 (m, 2H), 6.9 (dd, 1H, J = 2.4 Hz, J = 8.8 Hz), 4.85 (m, 2H), 4.38 (m, 1H), 4.1 (dd, 1H, J = 3.4 Hz, J = 11.0 Hz), 3.8 (s, 3H, OMe), 3.76 (dd, 1H, J = 9.4 Hz, J = 11.0 Hz, CH2-Cl); 13 C NMR (100 MHz, THF-d8) δ 161.6, 155.7, 147.6 (2C), 143.0, 141.3, 140.5, 138.0, 133.1, 132.1, 131.3, 129.9, 128.1, 127.4, 126.4, 125.9, 124.4 (2C), 120.0, 117.0, 113.6, 106.4, 103.0, 56.0, 55.8, 47.2, 44.3;

[0242] Example 34 - (1-(chloromethyl)-5-(4-hydroxyphenyl)-1H-benzo[e]indol-3(2H)-yl)(5-methoxybenzofuran-2-yl)methanone (ICT-11076) [ka]

[0243] Synthesized by Method B

[0244] 1 H NMR (400 MHz, DMSO-d6) δ 9.64 (s, 1H, NH), 8,29 (bs, 1H), 8.03 (d, 1H, J = 8.0 Hz), 7.84 (d, 1H, J = 8.4 Hz), 7.66 (s, 1H), 7.75 (d, 1H, J = 9.2 Hz), 7.58 (d, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.0 Hz), 7.54 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.4 Hz), 7.30 (d, 1H, J = 2.6 Hz), 7.27 (d, 2H, J = 8.4 Hz), 7.10 (dd, 1H, J = 2.6 Hz, J = 9.2 Hz), 6.92 (d, 2H, J = 8.4 Hz), 4.82 (dd, 1H, J = 10.4 Hz, J = 11.6 Hz), 4.66 (dd, 1H, J = 1.6 Hz, J = 11.6 Hz), 4.41 (m, 1H), 4.10 (dd, 1H, J = 3.2 Hz, J = 11.2 Hz), 3.99 (dd, 1H, J = 7.2 Hz, J = 11.2 Hz), 3.82 (s, 3H); 13 C NMR (100 MHz, DMSO-d6) δ 157.27, 157.05, 156.17, 149.31, 149.26, 141.10, 140,87, 130.83 (2C), 130.55, 129.69, 129.07, 127.43, 127.02, 126.70, 124.93, 124.42, 123.60, 117.79, 117.66, 115.32 (2C), 112.80, 112.69, 104.08, 55.64, 54.24, 47.46, 41.49; HRMS (ESI+) is C 29 H 23ClNO4484.13156 [M+H] + The result was 484.12993 calcd.

[0245] Example 35 - (1-(chloromethyl)-5-(4-hydroxyphenyl)-1H-benzo[e]indol-3(2H)-yl)(5-methoxybenzo[b]thiophen-2-yl)methanone (ICT-11077) [ka]

[0246] Synthesized by Method B

[0247] 1 H NMR (400 MHz, DMSO-d6) δ 9.65 (s, 1H, NH), 8.19 (bs, 1H), 8.08 (s, 1H), 8.03 (d, 1H, J = 8.2 Hz), 7.94 (d, 1H, J = 8.8 Hz), 7.85 (d, 1H, J= 8.0 Hz), 7.58 (dd, 1H, J = 7.4 Hz, J = 8.2 Hz), 7.53 (d, 1H, J = 2.4 Hz), 7.43 (dd, 1H, J = 7.4 Hz, J = 8.0 Hz), 7.26 (d, 2H, J = 8.0 Hz), 7.15 (dd, 1H, J = 2.4 Hz, J = 8.8 Hz), 6.92 (d, 2H, J = 8.0 Hz), 4.84 (dd, 1H, J = 9.6 Hz, J = 10.8 Hz), 4.56 (dd, 1H, J = 1.6 Hz, J = 10.8 Hz), 4.41 (m, 1H), 4.10 (dd, 1H, J = 3.2 Hz, J = 11.2 Hz), 4.02 (dd, 1H, J = 10.4 Hz, J = 11.2 Hz), 3.84 (s, 3H); 13C NMR (100 MHz, DMSO-d6) δ 161.12, 157.48, 157.05, 141.06, 140.91, 140.24, 139.96, 132.40, 130.85 (2C), 130.56, 129.74, HRMS (ESI+) is C 29 H 23 ClNO3S 500.10872 [M+H] + The result was 500.10809 calcd.

[0248] Example 36 - (1-(chloromethyl)-5-(4-hydroxyphenyl)-1,2-dihydropyrrolo[3,2-e]indol-3(6H)-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11084) [ka]

[0249] Synthesized by Method B

[0250] 1H NMR (400 MHz, CDCl3) δ 9.30 (s, 1H, NH), 8.51 (s, 1H), 8.37 (s, 1H), 7.56 (d, 2H, J= 8.4 Hz), 7.40 (d, 1H, J = 8.8 Hz), 7.33 (dd, 1H, J = 1.6 Hz,J = 2.8 Hz), 7.17 (d, 1H, J = 2.4 Hz), 7.06-7.02 (m, 4H), 6.73 (s, 1H), 6.59 (dd, 1H, J = 2.0 Hz, J = 3.2 Hz), 4.77 (m, 2H), 4.21 (m, 1H), 4.16 (dd, 1H, J = HRMS (ESI+) is C 27 H 22 ClN3NaO3494.12474 [M+Na] + The result was 494.12409 calcd.

[0251] Example 37 - (1-(chloromethyl)-5-(4-methoxyphenyl)-1,2-dihydropyrrolo[3,2-e]indol-3(6H)-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11085) [ka]

[0252] Synthesized by Method B

[0253] 1H NMR (400 MHz, CDCl3) δ 9.41(s,1H,NH), 8.51(s,1H), 8.36(s,1H), 7.60(d,2H,J=8.8Hz), 7.37(d,1H,J=8.9Hz), 7.32(dd,1H,J= 2.8 Hz, J = 3.2 Hz), 7.16 (d, 1H, J = 2.4 Hz), 7.06 (d, 2H, J = 8.8 Hz), 7.00 (dd, 1H, J = 2.4 Hz, J = 8.9 Hz), 6.79 (d, 1H, J = 1.6 Hz), 6.58 (dd, 1H, J = 1.6 Hz, J = 3.2 Hz), 4.76 (dd, 1H, J = 9.2 Hz, J = 10.8 Hz), 4.72 (dd, 1H, J = 4.0 Hz, J = 10.8 Hz), 4.20 (m, 1H), 4.14 (dd, 1H, J = 3.6 Hz, J). = 10.8 Hz), 3.90 (s, 3H), 3.89 (s, 3H), 3.66 (dd, 1H, J = 10.0 Hz, J = 10.8 Hz); 13 C NMR (100 MHz, CDCl3) δ 159.28, 154.64, 137.66, 132.08, 131.21, 131.16, 131.03, 129.47 (2C), 128.45, 126.08, 125.79. 123.93, 120.13, 116.37, 114.82, 114.64 (2C); HRMS (ESI+)C 28 H 25 ClN3O3486.15844 [M+H] + The 486.158833 calcd.

[0254] Example 38 - (1-(chloromethyl)-5-((4-methoxyphenyl)(methyl)amino)-1,2-dihydro-3H-benzo[e]indol-3-yl)(5-methoxy-1H-indol-2-yl)methanone (ICT-11067) [ka]

[0255] Synthesized by Method A

[0256] 1 H NMR (400 MHz, acetone-d6) δ 10.72 (s, 1H, NH), 8.42 (s, 1H), 8.0 (d, 1H, J = 8.0 Hz), 7.94 (d, 1H, J = 8.4 Hz), 7.56 (ddd, 1H,J = 1.6 Hz, J = 6.8 Hz, J = 8.0 Hz), 7.46 (d, 1H, J = 8.8 Hz), 7.36 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.4 Hz), 7.18 (m, 2H), 6.94 (dd, 1H, J = 2.4 Hz, J = 8.8 Hz), 6.77 (dd, 2H, J = 2.4 Hz, J = 6.8 Hz), 6.68 (dd, 2H, J = 2.4 Hz, J = 6.8 Hz), 4.87 (m, 2H), 4.43 (dd, 1H, J = 2.8 Hz, J= 8.4 Hz), 4.16 (dd, 1H, J = 3.2 Hz, J = 11.2 Hz), 3.94 (dd, 1H, J= 8.4 Hz, J = 11.2 Hz), 3.83 (s, 3H), 3.7 (s, 3H), 3.38 (s, 3H); 13C NMR (100 MHz, acetone-d6) δ 161.2, 155.5, 153.8, 148.7, 145.8, 143.8, 132.7, 132.0, 131.7, 129.2, 128.1, 126.0, 125.4, 124.4, 123.1, 117.6, 116.9, 116.5, 115.2, 113.8, 106.4, 103.0, 55.7, 47.7, 43.4, 41.7;

[0257] Example 39:

[0258] Compounds of the present invention were evaluated for their ability to inhibit the growth of MDA-MB-231 (triple-negative) and MCF-7 (estrogen receptor-positive) breast cancer cell lines. The compounds exhibited antiproliferative activity in the picomolar to micromolar range, as shown in Table 1. A positive control (PC), seco-CBI-MI (structure shown below), was used as a baseline for comparing the efficacy of compounds of the present invention. Surprisingly, it was found that modification of the phenolic OH group, previously thought to be crucial for efficacy, retained cellular activity. [ka] [Table 2]

[0259] The mechanism of action of naturally occurring duocarmycins is well documented and involves the spirocyclization of seco-duocarmycins, including those derived from CBI and other aryl-ring-fused duocarmycin alkylating subunits. Selected compounds (Example 15 (DP-31(CBI-MI)), Example 6 (DP36), Example 8 (DP45), Example 9 (DP47), Example 8 (DP45), Example 2 (DP59), Example 4 (DP65), and Example 5 (DP70)) generated DNA damage in the MDA-MB-231 human breast cancer cell line, as measured by H2AX phosphorylation as a marker of double-stranded DNA damage (Western blot images are shown in Figure 1). This observation suggests that modulation of the phenolic OH position of the CBI alkylating subunit reflects different pharmacodynamics and cytotoxicity via DNA damage.

[0260] The expression of γ-H2AX phosphorylation was investigated as a marker of DNA damage in MCF-7 human breast cancer cells after treatment with duocarmycin compounds. Breast cancer cells were treated with the duocarmycins of the present invention for 24 or 48 hours (concentration: 10×IC50) or with solvent alone (DMSO). γ-H2AX expression was determined by Western blot analysis and is shown in Figure 2.

[0261] This application provides novel substituents for use within the duocarmycin scaffold. The above examples demonstrate the functionality of the substituents of the invention as duocarmycins using a representative CBI core, the core shown in Formula IA. The substituents of the invention have been shown to be active as cancer therapeutics and are readily applicable to other core structures within the duocarmycin family. These cores are represented by compounds of Formulas IB-IL. Evidence for the activity of the cores of compounds of Formulas IB-IL can be found in the following review article and the references contained therein: "Chemical and Biological Explorations of the Family of CC-1065 and the Duocarmycin Natural Products," Ghosh et al., Current Topics in Medicinal Chemistry, 2009, Vol. 9, No. 16, 1494, which is incorporated herein by reference.

[0262] Example 42: Determination of antiproliferative activity of active duocarmycin drug leads in RMS cell lines

[0263] Briefly, 180 μL of cell suspension (approximately 2000 cells / well) was added to a new, sterile 96-well plate. Rhabdomyosarcoma (RMS) cell lines (RH30, RH41, and JR1) were then incubated at 37°C and 5% CO2 for 24 hours to allow cells to adhere to the well surface. The day after cell seeding, cells were treated with duocarmycin compounds. 20 μL of drug dilutions were added to the wells to a final volume of 200 μL / well. Untreated control wells received 20 μL of fresh medium (0.1% DMSO) instead. Stock concentrations of duocarmycin compounds were prepared in DMSO, but the latter concentration in wells containing medium and cells never exceeded 0.1% (v / v). The concentrations tested ranged from 100 μM to 1 pM, depending on the IC50 values ​​of each compound determined by preliminary screening. Each experiment was repeated at least three times, and antiproliferative activity was determined after 96 hours using a standard MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The data obtained are shown in Table 2 below. [Table 3]

[0264]

[0265] Further evidence of applicability to the cores found in formulas IB-IJ can be found in the following papers: pyrroloindolones representing the core of formula IB, DL Boger, DS Johnson, Angew. Chem., Int. Ed. Engl., 35 (1996), p. 1438; pyrazoloindolones representing the core of formula IC, PG Baraldi et al., Anti-Cancer Drug Des., 12 (1997), p. 67; cyclopropylfurano[e]indolones representing the core of formula ID, F. Mohamadi et al., J. Med. Chem., 37 (1994), p. 232, and Patel et al., Org. Chem., 62 (1997), p. 8868; 1,2,9,9a-tetrahydropyrido[3,2-e]indol-4-ones representing the core of formula IJ, Boger, DL; Boyce, CW, J. Org. Chem. 2000, 65, 4088-4100, and Core, representing formulas IK and IL, Mol. Cancer Ther. (2023) 22 (12), 1465-1478, all of which are incorporated herein by reference.

[0266] Example 43: Synthesis of modified payloads for conjugation to antibodies to form ADCs

[0267] Method for synthesizing AP-01 and AP-02Five equivalents of tert-butylmethyl(2-(methylamino)ethyl)carbamate (TBMMEC) (171.4 μL, 0.87 mmol) and 10 equivalents of DMAP (212.9 mg, 1.74 mmol) were dissolved in 4 mL of anhydrous CHCl and cooled to 0 °C under a nitrogen atmosphere. 2.5 equivalents of diphosgene (52.2 μL, 0.436 mmol) were added dropwise to the flask containing the amine, followed by stirring at room temperature for 30 min. In a separate flask, DP-36 (84 mg, 0.174 mmol) was dissolved in 2 mL of anhydrous THF under a nitrogen atmosphere. This solution was transferred to the first flask and allowed to react under a nitrogen atmosphere for 4 days. The reaction was quenched with cold water and stirred for 1 h. The product was extracted three times with CHCl. The organic phase was collected, dried, evaporated in vacuo and the crude product was purified by flash chromatography (hexane / ethyl acetate 1:1) to give the conjugate (85 mg, 70%) as a white solid.

[0268] tert-Butyl (4-(1-(chloromethyl)-3-(5-methoxy-1H-indole-2-carbonyl)-2,3-dihydro-1H-benzo[e]indol-5-yl)phenyl)ethane-1,2-diylbis(methylcarbamate) (AP-01) [ka]

[0269] 1H NMR (400 MHz, CDCl3) δ 9.56 (s, 1H); 8.53 (s, 1H); 7.92 (d, 1H, J = 8.4 Hz); 7.83 (d, 1H, J = 8.4 Hz); 7.55 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.4 Hz); 7.48 (d, 2H, J = 8.8 Hz); 7.37 (dd, 1H, J= 6.8 Hz, J = 8.4 Hz); 7.32 (d, 1H, J = 9.2 Hz); 7.23 (m, 2H); 7.12 (d, 1H, J = 2.4 Hz); 7.05 (dd, 1H, J = 1.2 Hz, J = 2.4 Hz); 6.96 (dd, 1H, J = 2.4 Hz, J = 9.2 Hz); 4.87 (dd, 1H, J= 1.6 Hz, J = 10.8 Hz); 4.71 (dd, 1H, J = 8.4 Hz, J = 10.8 Hz); 4.24 (m, 1H); 4.04 (dd, 1H, J = 3.2 Hz, J = 11.2 Hz); 3.86 (s, 3H); 3.66-3.47 (m, 5H); 3.17 + 3.08 (2s, 3H); 2.96 (m, 3H); 1.48 (s, 9H); ES + MS C 39 H 41 ClN4O6(696.2) m / z (%) 697.4 [M+H] + (20).

[0270] How to synthesize AP-03Boc-protected AP-01 (85 mg, 0.122 mmol) was dissolved in 7 mL of CHCl:TFA (6:1) on ice and stirred for 30 min. The solvent was removed under a stream of nitrogen on ice. The resulting product (72 mg, 0.122 mmol) was dried under vacuum to completely remove TFA, then dissolved in 6 mL of anhydrous DMF and 4 mL of anhydrous CHCl. ​​The mixture was reacted with Fmoc-Val-Ala-PAB-PNP (165 mg, 0.24 mmol) and DMAP (30 mg, 0.24 mmol) and stirred overnight at room temperature. The organic solvent was removed under reduced pressure, and the crude product was purified by flash column chromatography (CHCl → CHCl:MeOH 100:6) to give AP-03 (100 mg, 72%) as a white solid.

[0271] 4-((S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)propanamido)benzyl (4-(1-(chloromethyl)-3-(5-methoxy-1H-indole-2-carbonyl)-2,3-dihydro-1H-benzo[e]indol-5-yl)phenyl)ethane-1,2-diylbis(methylcarbamate) (AP-03) [ka]

[0272] 18.53 (s, 1H); 8.05 (d, 1H, J = 8.4 Hz); 7.84 (m, 3H); 7.71-7.65 (m, 4H); 7.59 (m, 1H); 7.52-7.27 (m, 11H); 7.22 (s, 1H); 7.18 (d, 1H, J = 3.2 Hz); 7.17-7.09 (m, 1H); 6.93 (dt, 1H, J = 2.4 Hz, J = 9.2 Hz); 6.7 (m, 1H); 5.13-5.02 (m, 2H); 4.96-4.87 (m, 2H); 4.6-4.52 (m, 1H); 4.49-4.43 (m, 1H); 4.41-4.28 (m, 2H); 4.21 (t, 1H, J = 6.8 Hz); 4.19-4.14 (m, 1H); 4.09 (m, 1H); 3.96 (dd, 1H, J = 8.4 Hz, J = 11.2 Hz); 3.82 (m, 3H); 3.68-3.51 (m, 4H); 3.14-2.96 (m, 6H); 2.15 (td, 1H, J = 0.8 Hz, J= 7.6 Hz); 1.33-1.19 (m, 3H); 0.92 (m, 6H); ES + MS C 65 H 64 ClNO 10 (1138.6) m / z (%) 1139.4 [M+H] + (100).

[0273] How to synthesize AP-04 AP-03 (100 mg, 0.088 mmol) was dissolved in 2 mL of THF, and 20 μL of DBU was added to the flask. The reaction mixture was stirred at room temperature for 3 minutes, neutralized with 1 mL of 95% acetic acid, and then lyophilized. The crude product was purified by column chromatography (CHCl:MeOH:TEA 90:10:1). Compound AP-04 (35 mg, 43%) was obtained as a white solid.

[0274] 4-((S)-2-((S)-2-amino-3-methylbutanamido)propanamido)benzyl (4-(1-(chloromethyl)-3-(5-methoxy-1H-indole-2-carbonyl)-2,3-dihydro-1H-benzo[e]indol-5-yl)phenyl)ethane-1,2-diylbis(methylcarbamate) (AP-04) [ka]

[0275] 1 H NMR (400 MHz, CDCl3) δ 8.97 (d, 1H, J = 11.2 Hz); 8.48 (s, 1H); 7.91 (m, 2H); 7.83 (d, 1H, J = 8 Hz); 7.57-7.48 (m, 3H); 7.42-7.21 (m, 6H); 7.14-7.09 (m, 2H); 7.06 (s, 1H); 7.0 (dd, 1H, J = 2.4 Hz, J = 9.2 Hz); 5.12-5.02 (m, 2H); 4.89 (d, 1H, J = 10.8 Hz); 4.73 (m, 1H); 4.64 (m, 1H); 4.25 (m, 4.05 (m, 1H); 3.87 (s, 3H); 3.7-3.47 (m, 5H); 3.3-2.98 (m, 7H); 2.27 (m, 1H); 1.31 (m, 3H); 0.96 (m, 3H); 0.82 (m, 3H);ES + MS C 50 H 54 ClN7O8(916.4) m / z(%) 917.3 [M+H] + (100).

[0276] How to synthesize AP-05Compound AP-04 (35 mg, 0.038 mmol) was dissolved in 1 mL of DMF, followed by the addition of DIPEA (16.6 μL, 0.095 mmol) and Mal-PEG4-NHS ester (29.4 mg, 0.057 mmol). The reaction was stirred at room temperature for 1 h. The DMF was removed under a stream of nitrogen and purified by semi-preparative HPLC according to the following method.

[0277] Semi-preparative HPLC purification method:

[0278] Solvent A: 90% water, 10% acetonitrile, 0.05% TFA

[0279] Solvent B: 90% acetonitrile, 10% water, 0.05% TFA [Table 4]

[0280] 4-(1-(chloromethyl)-3-(5-methoxy-1H-indole-2-carbonyl)-2,3-dihydro-1H-benzo[e]indol-5-yl)phenyl 4-((2S,5S)-25-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-5-isopropyl-2-methyl-4,7,23-trioxo-10,13,16,19-tetraoxa-3,6,22-triazapentacosanamide)benzyl ethane-1,2-diylbis(methylcarbamate) (AP-05) [ka]

[0281] 1H NMR (400 MHz, (CD3)2CO) δ 10.9 (bs, 1H); 9.22 (m, 1H); 8.53 (s, 1H); 8.07 (d, 1H, J = 8.4 Hz); 7.89-7.82 (m, 1H); 7.74-7.69 (m, 3H); 7.6 (ddd, 1H, J = 1.2 Hz, J = 6.8 Hz, J = 8.4 Hz); 7.52 (dd, 1H, J = 3.6 Hz, J = 9 Hz); 7.45-7.41 (m, 2H); 7.35 (m, 1H); 7.23 (bs, 1H); 7.19 (d, 1H, J = 2.8 Hz); 7.17-7.12 (m, 2H); 6.95 (dd, 1H, J= 2.4 Hz, J = 9 Hz); 6.81 (s, 2H); 5.18-5.02 (m, 2H); 4.97-4.88 (m, 2H); 4.52-4.45 (m, 2H); 4.31-4.25 (m, 1H); 4.21-4.15 (m, 1H); 3.97 (m, 1H); 3.83 (s, 3H); 3.73 (d, 2H, J = 7.6 Hz); 3.71 (d, 2H, J = 6.4 Hz); 3.69-3.51 (m, 16H); 3.47 (t, 2H, J = 5.6 Hz); 3.3 (q, 2H, J = 5.6 Hz); 3.08-2.98 (m, 6H); 2.55 (m, 2H); 2.45 (t, 2H, J = 7.4 Hz); 2.12 (m, 1H); 1.29-1.19 (m, 3H); 0.92 (m, 6H); ES + MS C 68 H 80 ClN9O 16 (1314.8) m / z (%) 680.1 [M+2Na] 2+ (100).

[0282] In the description and claims herein, the terms "comprise" and "contain" and variations thereof mean "including, but not limited to," and are not intended to exclude, and do not exclude, other moieties, additives, ingredients, integers, or steps. In the description and claims herein, the singular includes the plural unless the context dictates otherwise. In particular, where the indefinite article is used, the specification is to be interpreted as contemplating the plural as well as the singular, unless the context dictates otherwise.

[0283] It is understood that any feature, integer, property, compound, chemical moiety, or group described in connection with a particular aspect, embodiment, or example of the invention is also applicable to any other aspect, embodiment, or example described herein, unless inconsistent therewith. All features disclosed herein (including the accompanying claims, abstract, and drawings) and / or all steps of any method or process disclosed herein may be combined in any combination, except combinations in which at least some such features and / or steps are mutually exclusive. The invention is not limited to the details of any foregoing example. The invention extends to any novel one or any novel combination of features disclosed herein (including the accompanying claims, abstract, and drawings), or to any novel one or any novel combination of steps of any method or process disclosed herein.

[0284] The reader's attention is drawn to all papers and documents in connection with this application, filed contemporaneously with or prior to this application, and in the public domain herewith, and the entire contents of such papers and documents are incorporated herein by reference.

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt thereof: 【Chemistry 1】 During the ceremony: A represents a 5- or 6-membered ring, wherein there are 3 or 4 atoms not represented within said ring, said 3 or 4 atoms being selected as follows: 0, 1, or 2 of said atoms are independently selected from C═O, N, NH, S, or O, and the remaining atoms of said atoms consist of CH; X represents NH, S or O; Y represents a halo group; R １ Ha-OR ４ａ , -NR ４ａ R ４ｂ , substituted or unsubstituted C １－６ Alkyl, substituted or unsubstituted C ５－１０ Cycloalkyl, substituted or unsubstituted C ５－１０ Heterocycloalkyl, substituted or unsubstituted C ６－１０ Aryl, and substituted or unsubstituted C ５－１０ heteroaryl, where, when substituted, halo, -OR ４ａ , -NR ４ａ R ４ｂ , -NO ２ , C １－６ Alkyl, C １－６ Haloalkyl, C substituted with OH １－６ alkyl, C substituted with OMe １－６ C substituted with alkyl and ═O １－６ R is one, two, or three groups independently selected from alkyl １ the group is substituted; R ２ is H, C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, and —C(O)—R ５ represents a group selected from R ３ is H, -OH, halo, C １－６ Alkyl, C １－６ Haloalkyl, C １－６ Alkoxy, —NR ６ａ R ６ｂ , C ６－１０ Aryl-substituted C １－６ Alkoxy, and C ６－１０ Aryl-substituted C １－６ alkyl; R ４ａ and R ４ｂ are each independently １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, C substituted with OH １－６ alkyl, C substituted with OMe １－６ alkyl, C substituted with ═O １－６ Alkyl, substituted or unsubstituted C ６－１０ Aryl-substituted C １－６ Alkyl, —C(O)—C １－６ Alkyl, -(CR ７ａ R ７ｂ ) ｎ C(O)OR ８ , -(CR ７ａ R ７ｂ ) ｎ C(O)NHR ８ , substituted or unsubstituted C ５－１０ Cycloalkyl, substituted or unsubstituted C ５－１０ Heterocycloalkyl, substituted or unsubstituted C ６－１０ Aryl, and substituted or unsubstituted C ５－１０ heteroaryl; or R ４ｂ is H; where, if substituted, R ４ａ Groups and R ４ｂ The groups include halo, OH, —NO ２ , C １－６ Alkyl, C １－６ Alkoxy, C １－６ Haloalkyl, C substituted with OH １－６ alkyl, C substituted with OMe １－６ alkyl, C substituted with ═O １－６ Alkyl, and —NH ２ and is substituted with one or two groups selected from: n is 1, 2, or 3 (preferably 1); R ５ is C １－６ Alkyl, C ２－６ Alkenyl, and C ２－６ alkynyl; R ６ａ and R ６ｂ are each independently H and C １－６ alkyl; R ７ａ and R ７ｂ One of them is H, C １－６ C substituted with alkyl and —OH １－６ Alkyl, -SH, -SMe, -NH ２ , -C(O)OH, -C(O)NH ２ , guanidine, C ６－１０ Aryl, C substituted with —OH ６－１０ Aryl, or C ５－１０ heteroaryl; and all other R ７ａ and R ７ｂ is H; and R ８ is H and C １－６ alkyl; However, R ２ is methyl, R １ is not -OBn.

2. 2. The compound of claim 1, wherein the compound is a compound having a formula selected from the following: 【Chemistry 2】

3. 2. The compound of claim 1, wherein the compound is a compound of formula IA. 【Transformation 3】

4. R ３ is H, -OH, C １－６ Alkyl, C １－６ Alkoxy, —NR ６ａ R ６ｂ , and C ６－１０ Aryl-substituted C １－６ 10. A compound according to any preceding claim, which represents a group selected from: alkoxy.

5. R ３ is H, —OH, methyl, ethyl, —OMe, —NH ２ , -NHMe, -N(Me) ２ 5. The compound of claim 4, wherein the aryl group represents a group selected from -O, -O, and -OBn.

6. R ２ is C １－６ Alkyl and —C(O)—R ５ 10. A compound according to any preceding claim, wherein the compound represents a group selected from:

7. The compound according to one of claims 1 to 5, wherein the compound is a compound according to formula IIIa or IIIb. 【Chemistry 4】

8. The compound according to one of claims 1 to 5, wherein the compound is a compound according to formula VIIa or VIIb. 【Transformation 5】

9. R １ is -OR ４ａ , -NR ４ａ R ４ｂ , substituted or unsubstituted C ５－１０ Heterocycloalkyl, substituted or unsubstituted C ６－１０ Aryl, and substituted or unsubstituted C ５－１０ heteroaryl, where, if substituted, R １ The group is halo, OR ４ａ , -NR ４ａ R ４ｂ , -NO ２ , C １－６ alkyl, C substituted with OH １－６ C substituted with alkyl and ═O １－６ substituted with 1, 2, or 3 groups (optionally 1 or 2 groups) independently selected from alkyl; A compound according to any of the preceding claims.

10. R ４ａ and R ４ｂ are each independently C １－６ alkyl, C substituted with OH １－６ alkyl, C substituted with OMe １－６ Alkyl, C ６－１０ Aryl-substituted C １－６ Alkyl, -(CR ７ａ R ７ｂ ) ｎ C(O)OR ８ , -(CR ７ａ R ７ｂ ) ｎ C(O)NHR ８ , substituted or unsubstituted C ５－１０ Cycloalkyl, substituted or unsubstituted C ５－１０ Heterocycloalkyl, and substituted or unsubstituted C ６－１０ aryl; or R ４ｂ is H; where, if substituted, R ４ａ and R ４ｂ The groups include halo, OH, —NO ２ , C １－６ Alkyl, C １－６ Alkoxy, C １－６ Haloalkyl, C substituted with OH １－６ alkyl, C substituted with OMe １－６ alkyl, C substituted with ═O １－６ Alkyl, and —NH ２ (Optionally, halo, -NO ２ , C １－６ Alkyl, C １－６ Alkoxy, and C １－６ haloalkyl); A compound according to any of the preceding claims.

11. R ７ａ and R ７ｂ is H, C １－６ Alkyl, and —OH, —SH, —SMe, —NH ２ , -C(O)OH, -C(O)NH ２ , guanidine, benzene, phenol, indole or imidazole substituted C １－６ alkyl, and all other R ７ａ and R ７ｂ is H, A compound according to any of the preceding claims.

12. 10. A compound according to any preceding claim, wherein n is 1.

13. R １ is —OH (optionally, R ２ is not H), OBn, -N(Et) ２ , -NHBu, and a compound according to any preceding claim selected from: 【Transformation 6】

14. 2. The compound of claim 1, wherein said compound of formula I is selected from the following: 【Transformation 7】 【Transformation 8】 【Chemistry 9】 【Chemistry 10】 【Chemistry 11】 【Chemistry 12】

15. 10. A compound according to any preceding claim for use as a medicament.

16. 10. A compound according to any preceding claim for use in a method for the treatment of a condition treatable by administration of a cytotoxic agent.

17. 10. A compound according to any preceding claim for use in a method for the treatment of cancer.

18. 18. The compound of claim 17, wherein the cancer is selected from breast, lung, prostate, colon, bladder, brain, pancreas, head and neck, and neuroblastoma.

19. Use of a compound according to any one of claims 1 to 14 in the manufacture of an antibody drug conjugate.

20. An antibody-drug conjugate, wherein the drug is a compound according to any one of claims 1 to 14.

21. 21. The antibody drug conjugate of claim 20, further comprising an antibody, biosimilar, affimer, or other non-antibody binding protein, and a linker connecting the drug to the antibody.

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