Peptide drug conjugates

Abstract

The present invention provides a pharmaceutical formulation comprising negatively charged cyclodextrin or a pharmaceutically acceptable derivative of negatively charged cyclodextrin and a compound # imgabs0 # of formula (I) as specified in the application and the use of these formulations in the treatment of diseases such as cancer. Related compositions, kits, methods of treatment, and methods of manufacture are also provided. The invention is particularly suitable for treating hematologic cancer.

Description

Technical Field

[0001] The present invention relates to pharmaceutical formulations comprising a negatively charged cyclodextrin or a derivative thereof and a peptide drug conjugate (PDC) compound and the use of these formulations in treating diseases such as cancer. Background Art

[0002] It has been found that compounds of formula (I) as defined below are effective anticancer agents. In particular, compounds of formula (I) exhibit excellent in vitro cytotoxicity against various hematological cancer cell lines. Compounds of formula (I) are also effective in reducing tumor growth in an in ovo chicken embryo xenograft model of lymphoma. Furthermore, compounds of formula (I) are readily hydrolyzed within cancer cells to form metabolites that are preferentially sequestered and retained within the cells and have strong alkylating activity.

[0003] There remains a need to provide a stable pharmaceutical formulation comprising a compound of formula (I) and a pharmaceutically acceptable salt, ester, amide or carbamate thereof, as well as salts of such esters, amides or carbamates. Summary of the Invention

[0004] The present invention provides a pharmaceutical formulation comprising a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin and a compound according to formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate.

[0005] The present invention also provides a composition comprising the pharmaceutical preparation according to the present invention and a physiologically acceptable aqueous solvent or diluent.

[0006] The present invention also provides a kit comprising the pharmaceutical formulation according to the present invention and a physiologically acceptable aqueous solvent or diluent.

[0007] The present invention also provides a pharmaceutical formulation, composition or kit according to the invention for use as a medicament.

[0008] The present invention also provides a pharmaceutical preparation, composition or kit according to the present invention, which is used for treating or preventing cancer.

[0009] The present invention also provides a method for treating a patient, comprising administering a pharmaceutically effective amount of the pharmaceutical preparation or composition according to the present invention.

[0010] The present invention also provides a method for treating or preventing cancer, which comprises administering an effective amount of a pharmaceutical preparation or composition according to the present invention, for example, wherein the cancer is any one of the following: blood cancer / blood cell cancer, such as leukemia (e.g., acute lymphocytic leukemia, including adult and childhood acute lymphocytic leukemia; acute myeloid leukemia, including adult and childhood acute myeloid leukemia; chronic lymphocytic leukemia, such as B-cell chronic lymphocytic leukemia; chronic myelogenous leukemia; and hairy cell leukemia); lymphoma (e.g., AIDS-related lymphoma; cutaneous T-cell lymphoma; Hodgkin's lymphoma, including adult and childhood Hodgkin's lymphoma and pregnancy Hodgkin's lymphoma; non-Hodgkin's lymphoma, including adult and childhood non-Hodgkin's lymphoma and pregnancy non-Hodgkin's lymphoma; mycosis fungoides; Sezary syndrome; Waldenstrom's macroglobulinemia; macroglobulinemia); primary mediastinal large B-cell lymphoma; mantle cell lymphoma; diffuse large B-cell lymphoma; and primary central nervous system lymphoma); and other hematologic cancers (e.g., chronic myeloproliferative disorders; multiple myeloma / plasma cell neoplasms; myelodysplastic syndromes; and myelodysplastic / myeloproliferative disorders); osteosarcoma; ovarian cancer; breast cancer; lung cancer; glioblastoma; retinoblastoma; and metastases of the above cancers.

[0011] The present invention further provides a composition (e.g., a pharmaceutical composition) as described above or a kit as described above for use in treating and / or preventing blood cancers, such as blood cancers and related clonal disorders (e.g., MGUS or amyloidosis). Blood cancers can include, for example, plasma cell neoplasms and myelomas (e.g., MGUS, plasma cell neoplasms, smoldering myeloma, myeloma), multiple myeloma, relapsed / refractory multiple myeloma, light-chain myeloma or non-secretory myeloma, and plasma cell leukemia); B-cell leukemias (e.g., acute lymphoblastic leukemia, including adult and childhood acute lymphoblastic leukemia; chronic lymphocytic leukemia; and hairy cell leukemia) and B-cell-derived lymphoid malignancies (e.g., AIDS-related lymphoma; Hodgkin's lymphoma, including adult and childhood Hodgkin's lymphoma and pregnancy Hodgkin's lymphoma; non-Hodgkin's lymphoma, including adult and childhood non-Hodgkin's lymphoma and pregnancy non-Hodgkin's lymphoma; Waldenstrom's macroglobulinemia; primary mediastinal large B-cell lymphoma; diffuse large B-cell lymphoma; follicular lymphoma; mantle cell lymphoma; hairy cell lymphoma; and primary central nervous system lymphoma).

[0012] Hematologic cancers can include, for example, plasma cell neoplasms and myeloma (e.g., multiple myeloma or relapsed / refractory multiple myeloma), B cell leukemias (e.g., chronic lymphocytic leukemia (CLL), acute lymphocytic leukemia, or hairy cell leukemia), and B cell-derived lymphoid malignancies (e.g., diffuse large B cell lymphoma; follicular lymphoma; mantle cell lymphoma; hairy cell lymphoma; and primary central nervous system lymphoma). Hematologic cancers can, for example, be selected from the group consisting of: plasma cell neoplasms or myeloma, B cell leukemia, or B cell-derived lymphoid malignancies, such as multiple myeloma, chronic lymphocytic leukemia (CLL), or diffuse large B cell lymphoma. In one embodiment of the invention, the hematologic cancer is multiple myeloma (e.g., relapsed / refractory multiple myeloma) or diffuse large B cell lymphoma, such as multiple myeloma or, for example, diffuse large B cell lymphoma.

[0013] The present invention also provides the use of a pharmaceutical preparation, composition or kit according to the present invention for the manufacture of a medicament for treating or preventing cancer, for example, wherein the cancer is any one of the following: a blood cancer / blood cell cancer, such as a leukemia (e.g., acute lymphocytic leukemia, including adult and childhood acute lymphocytic leukemia; acute myeloid leukemia, including adult and childhood acute myeloid leukemia; chronic lymphocytic leukemia, such as B-cell chronic lymphocytic leukemia; chronic myeloid leukemia; and hairy cell leukemia); a lymphoma (e.g., AIDS-related lymphoma; cutaneous T-cell lymphoma; Hodgkin's lymphoma, including adult and childhood Hodgkin's lymphoma); The present invention includes the following types of cancer: Hodgkin's lymphoma, including adult and pediatric non-Hodgkin's lymphoma and non-Hodgkin's lymphoma during pregnancy; non-Hodgkin's lymphoma, including adult and pediatric non-Hodgkin's lymphoma and non-Hodgkin's lymphoma during pregnancy; mycosis fungoides; Sézary syndrome; Waldenstrom's macroglobulinemia; primary mediastinal large B-cell lymphoma; mantle cell lymphoma; diffuse large B-cell lymphoma; and primary central nervous system lymphoma); and other hematologic cancers (e.g., chronic myeloproliferative disorders; multiple myeloma / plasma cell neoplasms; myelodysplastic syndromes; and myelodysplastic / myeloproliferative disorders); osteosarcoma; ovarian cancer; breast cancer; lung cancer; glioblastoma; retinoblastoma; and metastases of the above cancers. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 shows a metabolite compound (Exemplary Compound 18) identified after treatment with Exemplary Compound 1 as described herein ( Figure 1A ) and bendamustine ( Figure 1A ) and melphalan ( Figure 1B ) Intracellular and extracellular concentrations of the comparators after treatment of MM.1S cells in vitro.

[0015] Figure 2Shown are the effects of exemplary Compound 1 (left panel) and bendamustine (right panel) on the growth of xenograft tumors initiated by the SU-DHL-4 human lymphoma cell line in an in ovo chick embryo model.

[0016] Figures 3(A) to 3(G) Shown are the results of measurements of intracellular and extracellular concentrations of the compounds described herein and their metabolites following in vitro treatment of MM.1S cells.

[0017] Figure 4 (A) to 4(C) show the results of experiments evaluating DNA damage caused by compounds described herein and controls.

[0018] Figures 5(A) to 5(B) Shown are the results of an assay evaluating DNA fragmentation caused by compounds described herein and controls. Figure 5B Representative FACS histograms of Alexa fluor 488 intensity following treatment with a compound of formula (I) (upper panel), bendamustine (middle panel), or a control (lower panel) are shown. DETAILED DESCRIPTION

[0019] The present invention provides a pharmaceutical formulation comprising a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin and a compound according to formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate.

[0020] The present invention has found that pharmaceutical formulations comprising a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin and a compound according to formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate are unexpectedly stable.

[0021] Compound of formula (I)

[0022] The compounds of formula (I) are a new class of PDCs that are highly cytotoxic to human cancer cells, particularly human blood cancer cells. These have been disclosed in detail in international patent application number PCT / EP2022 / 066756, which was published as WO2022 / 263679.

[0023] The preferred compounds described in International Patent Application No. PCT / EP2022 / 066756 are also preferred compounds for use in the formulations, compositions, kits, methods of treatment, and uses claimed herein. Preferably, the compound of formula (I) or its ester, amide, or carbamate is present in the form of a pharmaceutically acceptable salt thereof. Most preferably, the compound of formula (I) or its ester, amide, or carbamate is present in the form of a hydrochloride salt.

[0024] PCT / EP2022 / 066756

[0025] A new type of PDC has been found, which is highly cytotoxic to human cancer cells, particularly human blood cancer cells. As described in the Examples section (including a cross-reference to PCT application number PCT / EP2022 / 066756 published as numbered WO2022 / 263679), various example compounds as described herein have been synthesized and their cytotoxicity to several blood cancer cell lines has been tested in an in vitro cytotoxicity assay. It has been found that compounds as described herein have strong efficacy and show selectivity to blood cancer cells, as shown by the lower cytotoxicity of these compounds to the fibroblast cell line BJ. Example compound 1 has been found to be particularly effective for inhibiting tumor growth in an in-ovo chicken embryo xenograft model using a human lymphoma cell line SU-DHL-4.

[0026] In further studies, it has been found that the exemplary compounds described herein are easily hydrolyzed in MM.1S cancer cells to form metabolites that are sequestered and retained in the cells and have strong alkylating activity, thereby proving that the compounds described herein are a new class of effective PDCs for treating or preventing cancer, especially for treating or preventing blood cancer.

[0027] Described herein are compounds of formula (I):

[0028]

[0029] W1, W2, W3 and W4 are each CH, or one of W1, W2, W3 and W4 is N and the rest are CH.

[0030] In one embodiment, W2, W3 and W4 are each CH, and W1 is N.

[0031] It is easy to see that the –N(CH2CH2Cl)2 group is attached to the core of the molecule via a carbon atom in the ring. For the corresponding W group, the H in the CH group is correspondingly absent.

[0032] In a preferred embodiment, W1, W2, W3 and W4 are each CH, so that formula (I) has formula (Ia):

[0033]

[0034] In a preferred embodiment, formula (I) is a compound of formula (Ib),

[0035]

[0036] In the compound of formula (I), X is C 1-6 For example, X can be C 1-4 Alkylene, C 1-3 Alkylene, C1-2 Alkylene, C2 alkylene or C1 alkylene. X can be a straight chain or branched chain alkylene. The alkylene at position X is present in the imidazole portion and the peptide portion of the compound described herein (i.e., R 3 The linkage is formed between the alkylene groups (i.e., -CH2-) and the alkylene groups (i.e., -CH2-)). It has been found that alkylene linking groups of one or two carbons in length at the X position are particularly effective. Thus, in a preferred embodiment, X is a C1 alkylene group (i.e., -CH2-) or a straight-chain C2 alkylene group (i.e., -CH2-CH2-).

[0037] In the compound of formula (I), R 1 is selected from the group consisting of: H; C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-4 Alkyl; and halogen (eg, selected from the group consisting of: H; C 1-4 alkyl; and halogen). For example, R 1 Can be selected from the group consisting of: H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, F, Cl, Br and I. Optionally, these groups can be substituted by 1, 2 or 3 groups independently selected from halogen. In certain embodiments, R 1 is selected from the group consisting of: H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, F and Cl. In certain preferred embodiments, R 1 For H.

[0038] In the compound of formula (I), R 2 is selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and -C 1-6 Alkyl. For example, R 2 may be selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 F or Cl; and -C 1-4 In one embodiment, R 2 is selected from the group consisting of: phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and -C 1-6 Alkyl, for example -C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-4 In certain embodiments, R 2 For unsubstituted -C 1-2 an alkyl group (eg, methyl group) or an unsubstituted phenyl group.

[0039] In the compound of formula (I), R 3 is a group according to formula (II):

[0040]

[0041] or a group according to formula (III):

[0042]

[0043] In a preferred embodiment, R 3 is a group according to formula (IIa):

[0044]

[0045] or a group according to formula (IIIa):

[0046]

[0047] For the avoidance of doubt, in formulas (II), (III), (IIa) and (IIIa) represents the point of attachment of Formula (II), (III), (IIa) or (IIIa) to Formula (I), (Ia) or (IIa).

[0048] In formulas (II), (IIa), (III) and (IIIa), R 4 Selected from the group consisting of: N(R c )(R d ) and formula (IV):

[0049]

[0050] In a preferred embodiment, R 4 Selected from the group consisting of: N(R c )(R d ) and formula (IVa):

[0051]

[0052] For the avoidance of doubt, in formulas (IV) and (IVa) represents the point of attachment of formula (IV) or (IVa) to formula (II), (III), (IIa) or (IIIa).

[0053] In the compound of formula (I), when R 4 When it is formula (IV) or (IVa), R 5 R b .

[0054] R b Selected from the group consisting of: -OH-N(Re )(R f ); and -OC optionally substituted by one or more groups selected from the following 1-6 Alkyl: halogen, -OH, -CN, -N(R e )(R f ),-C 6-10 Aryl or a 3- to 12-membered heterocyclic ring containing one or more O, N or S atoms and optionally substituted by 1, 2 or 3 halogens, and / or wherein the alkyl group is optionally interrupted by 1, 2 or 3 O, N or S atoms.

[0055] For example, when R 4 When it is formula (IV) or (IVa), R 5 (It is R b ) may be selected from the group consisting of: -OH and -OC 1-4 Preferably, when R 4 When it is formula (IV) or (IVa), R 5 Selected from the group consisting of: -OH and -OC 1-3 More preferably, when R 4 When it is formula (IV) or (IVa), R 5 Selected from the group consisting of methoxy, ethoxy or isopropoxy.

[0056] In the compound of formula (Ia), when R 4 N(R c )(R d ), R 5 Selected from the group consisting of: R b and formula (V):

[0057]

[0058] In a preferred embodiment, when R 4 N(R c )(R d ), R 5 -OH, -OC 1-6 Alkyl or formula (Va):

[0059]

[0060] For the avoidance of doubt, in formulas (V) and (Va) represents the point of attachment of Formula (V) or (Va) to Formula (II), (III), (IIa) or (IIIa).

[0061] In R4 N(R c )(R d ), R 5 (ie R b ) can be selected from the group consisting of: -OH, -OC 1-4 Alkyl (e.g., methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, or tert-butoxy), or R 5 Can be selected from formula (V) and formula (Va). Preferably, when R 4 N(R c )(R d ), R 5 Selected from the group consisting of: -OH, -OC 1-3 More preferably, when R 4 N(R c )(R d ), R 5 Selected from the group consisting of methoxy, ethoxy or isopropoxy.

[0062] It has been found that compounds of formula (I) (wherein R 4 N(R c )(R d ), formula (IV) or (IVa), and R 5 (ie R b ) is -OC 1-3 Alkyl groups (e.g., methoxy, ethoxy, or isopropoxy)) are particularly cytotoxic to cancer cells. Therefore, in certain preferred embodiments, in the compounds of formula (I), R 5 For-OC 1-3 That is, when R 4 N(R c )(R d ), formula (IV) or (IVa), R 5 Preferably -OC 1-3 Alkyl (eg, methoxy, ethoxy, or isopropoxy).

[0063] In certain embodiments, R 5 It is -OH.

[0064] In the compound of formula (I), each R a Independently selected from the group consisting of: H; C 1-6 Alkyl; -CH2-phenyl; or -CH2-3 to 12-membered heterocyclic group containing 1, 2, 3 or 4 heteroatoms selected from N, O and S; wherein said C 1-6The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of: -OH, -OC 1-6 alkyl, -NH2, -NHC(=NH)NH2, -C(O)OH, -C(O)NH2, -SH, -SCH3 and halogen; and the phenyl or heterocyclic group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halogen, -NH2, -OH, -OC 1-6 In certain embodiments, each R a Independently selected from the group consisting of: H; -C 1-6 Alkyl; -CH2-indolyl; -CH2-phenyl; and -CH2-5-membered heteroaryl containing 1, 2, 3 or 4 N or S atoms; wherein C 1-6 The alkyl group is optionally substituted with -OH; -NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen (e.g., F, Cl, Br, or I); and the phenyl group is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of halogen (e.g., F, Cl, Br, or I); -NH2; -OH; -OC 1-6 alkyl; and -NO2. Preferably, each R a Independently selected from the group consisting of: H; C 1-4 alkyl; and -CH2-phenyl; wherein the alkyl or phenyl group is optionally substituted with 1 or 2 halogens (eg, F or Cl). For example, each R a R is independently methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, -CH2-phenyl, -CH2-fluorophenyl, -CH2-chlorophenyl, -CH2-difluorophenyl or -CH2-dichlorophenyl. a It may independently be isopropyl, isobutyl, sec-butyl, -CH2-phenyl or -CH2-fluorophenyl (ie, 2-fluorobenzyl, 3-fluorobenzyl or 4-fluorobenzyl).

[0065] In the compound of formula (I), R b Selected from the group consisting of: -OH; -N(R e )(R f ); and -OC optionally substituted by one or more groups selected from the following 1-6 Alkyl: halogen, -OH, -CN, -N(R e )(R f ),-C 6-10 In certain embodiments, Rb -OH or -OC 1-6 wherein said alkyl group is optionally interrupted by 1, 2, or 3 O or N atoms. In certain other embodiments, R b -OH or -OC 1-6 Alkyl groups, such as -OC 1-6 Preferably, R b For-OC 1-4 Alkyl. For example, R b It can be methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy or sec-butoxy. In certain preferred embodiments, R b is methoxy, ethoxy or isopropoxy. More preferably, R b For ethoxy.

[0066] In certain embodiments, R b It is -OH.

[0067] R c and R d Each independently selected from the group consisting of: H, -C 1-6 Alkyl, -C(O)C 1-6 Alkyl and -CH2-phenyl, wherein the alkyl or the phenyl is optionally substituted by 1, 2 or 3 groups selected from halogen. In preferred compounds, R c is H, and R d Selected from H, -C 1-6 Alkyl and -C(O)C 1-6 Alkyl, wherein the alkyl is optionally substituted by 1, 2 or 3 groups selected from halogen. For example, R c is H, and R d Selected from H, -C 1-4 Alkyl and C(O)C 1-4 Alkyl. For example, R c H, C and R d is selected from H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, -C(O)methyl, -C(O)ethyl, -C(O)propyl, -C(O)isopropyl, -C(O)butyl, -C(O)isobutyl and -C(O)sec-butyl. Preferably, R c is H, and R d is selected from H, methyl and -C(O)methyl. Preferably, R c is H, and R d For H.

[0068] In the compound of formula (I), R e and R f Each independently selected from the group consisting of: H and -C 16Alkyl, wherein the alkyl is optionally substituted with 1, 2 or 3 groups selected from halogen; or R e and R f Together with the nitrogen atom to which they are attached, they form a 4-, 5- or 6-membered heterocyclic ring which is optionally substituted by 1, 2 or 3 groups selected from halogen. e and R f Each is independently selected from the group consisting of H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl and sec-butyl. e and R f are each independently selected from the group consisting of: H and methyl. Preferably, R e and R f All are H, all are methyl or R e and R f One of the is H and the other is methyl.

[0069] In certain embodiments, the compounds described herein are of Formula (Ia), wherein

[0070] X is -C 1-2 Alkylene (i.e., -CH2- or -CH2-CH2-);

[0071] R 1 is H;

[0072] R 2 is -C optionally substituted by 1, 2 or 3 F 1-2 Alkyl or phenyl optionally substituted by 1, 2 or 3 F, for example, R 2 Can be unsubstituted -C 1-2 Alkyl or unsubstituted phenyl;

[0073] R 3 is formula (IIa) or (IIIa);

[0074] R 4 Selected from the group consisting of: N(R c )(R d ) and formula (IVa);

[0075] When R 4 When it is formula (IVa), R 5 (ie R b ) is -OC 1-4 Alkyl; and when R 4 N(R c )(R d ), R 5 For-OC 1-4 Alkyl or formula (Va);

[0076] Each R aIndependently selected from the group consisting of: H; -C 16 Alkyl; CH2-indolyl; CH2-phenyl; or -CH2-5-membered heteroaryl containing 1, 2, 3 or 4 N or S atoms; wherein the -C 1-6 The alkyl group is optionally substituted with OH; NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and the phenyl group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halogen; -NH2; -OH; -OC 1-6 Alkyl; and -NO2;

[0077] R b For-OC 1-6 alkyl; and

[0078] R c is H, and R d Selected from H, -C 1-4 Alkyl and -C(O)C 1-4 alkyl;

[0079] or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including salts of such esters, amides or carbamates.

[0080] In certain other embodiments, the compounds described herein are of Formula (Ia), wherein

[0081] X is -C 1-2 Alkylene (i.e., -CH2- or -CH2-CH2-);

[0082] R 1 is H;

[0083] R 2 is methyl or phenyl;

[0084] R 3 is formula (IIa) or (IIIa);

[0085] R 4 N(R c )(R d ) or formula (IVa);

[0086] R 5 (ie R b ) is -OC 1-3 alkyl; and

[0087] Each R a Independently selected from the group consisting of: H; -C 16Alkyl; CH2-indolyl; CH2-phenyl; or -CH2-5-membered heteroaryl containing 1, 2, 3 or 4 N or S atoms; wherein C 1-6 The alkyl group is optionally substituted with OH; NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and the phenyl group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halogen; -NH2; -OH; -OC 1-6 Alkyl; and -NO2;

[0088] or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including salts of such esters, amides or carbamates.

[0089] It has been found that the compounds described herein are particularly cytotoxic to hematological cancer cells in in vitro cytotoxicity assays and in an in ovo chick embryo xenograft model of lymphoma when they are compounds of formula (Ia) wherein:

[0090] X is -C 1-2 Alkylene (i.e., -CH2- or -CH2-CH2-);

[0091] R 1 is H;

[0092] R 2 is methyl or phenyl;

[0093] R 3 is formula (IIa) or (IIIa);

[0094] R 4 N(R c )(R d ) or formula (IVa);

[0095] R 5 (ie R b ) is -OC 1-3 alkyl; and

[0096] Each R a independently selected from the group consisting of isopropyl, isobutyl, and -CH2-phenyl optionally substituted with 1 F (e.g., 2-fluorobenzyl, 3-fluorobenzyl, or 4-fluorobenzyl);

[0097] or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including salts of such esters, amides or carbamates.

[0098] In certain other embodiments, the compounds described herein are of Formula (Ia), wherein

[0099] X is -C 1-2Alkylene (i.e., -CH2- or -CH2-CH2-);

[0100] R 1 is H;

[0101] R 2 is -C optionally substituted by 1, 2 or 3 F 1-2 Alkyl or phenyl optionally substituted by 1, 2 or 3 F, for example, R 2 Can be unsubstituted -C 1-2 Alkyl or unsubstituted phenyl;

[0102] R 3 is formula (IIa);

[0103] R 4 is NH2;

[0104] R 5 (ie R b ) is -OC 1-4 alkyl; and

[0105] Each R a Independently selected from the group consisting of: H; -C 16 Alkyl; CH2-indolyl; CH2-phenyl; or -CH2-5-membered heteroaryl containing 1, 2, 3 or 4 N or S atoms; wherein C 1-6 The alkyl group is optionally substituted with OH; NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and the phenyl group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halogen; -NH2; -OH; -OC 1-6 Alkyl; and -NO2;

[0106] or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including salts of such esters, amides or carbamates.

[0107] In certain other embodiments, the compounds described herein are of Formula (Ia), wherein

[0108] X is -C 1-2 Alkylene (i.e., -CH2- or -CH2-CH2-);

[0109] R 1 is H;

[0110] R 2 is -C optionally substituted by 1, 2 or 3 F 1-2 Alkyl or phenyl optionally substituted by 1, 2 or 3 F, for example, R 2 Can be unsubstituted -C 1-2Alkyl or unsubstituted phenyl;

[0111] R 3 is formula (IIIa);

[0112] R 4 is NH2;

[0113] R 5 For-OC 1-4 alkyl; and

[0114] Each R a Independently selected from the group consisting of: H; -C 16 Alkyl; CH2-indolyl; CH2-phenyl; or -CH2-5-membered heteroaryl containing 1, 2, 3 or 4 N or S atoms; wherein C 1-6 The alkyl group is optionally substituted with OH; NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and the phenyl group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halogen; -NH2; -OH; -OC 1-6 Alkyl; and -NO2;

[0115] or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including salts of such esters, amides or carbamates.

[0116] In certain other embodiments, the compounds described herein are of Formula (Ia), wherein

[0117] X is -C 1-2 Alkylene (i.e., -CH2- or -CH2-CH2-);

[0118] R 1 is H;

[0119] R 2 is -C optionally substituted by 1, 2 or 3 F 1-2 Alkyl or phenyl optionally substituted by 1, 2 or 3 F, for example, R 2 Can be unsubstituted -C 1-2 Alkyl or unsubstituted phenyl;

[0120] R 3 is formula (IIIa);

[0121] R 4 is formula (IVa);

[0122] R 5 For-OC 1-4 alkyl; and

[0123] Each R aIndependently selected from the group consisting of: H; -C 16 Alkyl; CH2-indolyl; CH2-phenyl; or -CH2-5-membered heteroaryl containing 1, 2, 3 or 4 N or S atoms; wherein -C 1-6 The alkyl group is optionally substituted with OH; NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and the phenyl group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halogen; -NH2; -OH; -OC 1-6 Alkyl; and -NO2;

[0124] or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including salts of such esters, amides or carbamates.

[0125] In certain preferred embodiments, the compound is:

[0126] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 1);

[0127] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-(4-fluorophenyl)propanoic acid ethyl ester (Example Compound 2);

[0128] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-methyl-butyric acid ethyl ester (Example Compound 3);

[0129] (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 4);

[0130] (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-3-(4-fluorophenyl)propionic acid ethyl ester (Example Compound 5);

[0131] (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-3-methyl-butyric acid ethyl ester (Example Compound 6);

[0132] (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 7);

[0133] (2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propionyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Exemplary Compound 8);

[0134] (2S)-2-[[(2S)-2-amino-3-methyl-butyryl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 9);

[0135] (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionic acid ethyl ester (Example Compound 10);

[0136] (2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propionyl]amino]-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionic acid ethyl ester (Example Compound 11);

[0137] (2S)-2-[[(2S)-2-amino-3-methyl-butyryl]amino]-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionic acid ethyl ester (Example Compound 12);

[0138] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-phenyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 13);

[0139] (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid methyl ester (Example Compound 14);

[0140] (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid isopropyl ester (Exemplary Compound 15);

[0141] (2S)-2-[[(2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propionyl]amino]-4-methyl-pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 16);

[0142] (2S)-2-[[(2R)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propionyl]amino]-4-methyl-pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 21)

[0143] (2S)-2-[[(2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoyl]amino]-3-(4-fluorophenyl)propionic acid ethyl ester (Example Compound 22)

[0144] (2S)-2-[[(2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 23)

[0145] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid 2-morpholinoethyl ester (Example Compound 24)

[0146] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid 2-isopropoxyethyl ester (Example Compound 25)

[0147] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-valeric acid isopropyl ester (Example Compound 26)

[0148] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid methyl ester (Example Compound 27)

[0149] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid 3-(dimethylamino)propyl ester (Example Compound 28)

[0150] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (2-methoxy-1-methyl-ethyl ester) (Example Compound 29)

[0151] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-N,N,4-trimethyl-pentanamide (Example Compound 30)

[0152] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-N,4-dimethyl-pentanamide (Example Compound 31)

[0153] (2S)-2-[[(2S)-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]-2-(methylamino)butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 32)

[0154] (2S)-2-[[(2S)-2-acetylamino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-(4-fluorophenyl)propionic acid ethyl ester (Example Compound 34)

[0155] (2S)-2-[[(2S)-2-Acetylamino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 35)

[0156] (2S)-2-[[(2S)-2-acetylamino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-3-(4-fluorophenyl)propionic acid ethyl ester (Example Compound 36)

[0157] (2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 38)

[0158] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 45)

[0159] (2S)-2-[[(2R)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 46)

[0160] (2R)-2-[[(2R)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 47)

[0161] (2S)-2-[[(2S)-2-amino-4-[6-[bis(2-chloroethyl)amino]-3-methyl-imidazo[4,5-b]pyridin-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Exemplary Compound 48); and

[0162] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloro-1,1,2,2-tetradeuterium-ethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 50)

[0163] or a pharmaceutically acceptable salt, amide or carbamate thereof, including salts of such amides or carbamates.

[0164] The compounds described herein can be prepared using methods known to those skilled in the art of organic chemistry. Exemplary procedures for preparing compounds of formula (I) are described in the Examples section (including a cross-reference to PCT application number PCT / EP2022 / 066756, published as WO2022 / 263679).

[0165] In embodiments, the compounds described herein may contain isotopic atoms. As defined herein, an isotopic atom is an atom of an element that is not the most common naturally occurring isotope. Deuterium is a safe and stable isotope of hydrogen. In one embodiment, the deuterium abundance level of the compounds described herein is greater than the naturally occurring deuterium abundance. The naturally occurring deuterium abundance is 0.0156 mol%, where mol% is the percentage of deuterium to the total moles of hydrogen in a sample. Thus, in one mole of naturally occurring hydrogen, 0.156 mmol is deuterium, or in one mole of naturally occurring hydrogen, 6.022 x 10 23 In a sample of naturally occurring hydrogen atoms, 9.39×10 19Deuterium atoms, or in a sample with 6413 naturally occurring hydrogen atoms, there is one deuterium atom. The deuterium abundance level higher than the naturally occurring deuterium abundance can be at least 1 mol%, 5 mol%, 10 mol%, 50 mol%, 90 mol% or 98 mol% deuterium. In certain embodiments, the deuterium abundance level of the compounds described herein is at least 1 mol%, 5 mol%, 10 mol%, 50 mol%, 90 mol% or 98 mol% deuterium. Procedures for preparing deuterated compounds are known in the art. See, for example, Sajiki, New Horizons of Process Chemistry (2017), Springer, pages 29-40 and Hanson, The Organic Chemistry of Isotopic Labelling (2011), Chapter 3, RSC Publishing.

[0166] Depending on the substituents present in the compounds described herein, the compounds may form esters, amides, carbamates, and / or salts. Salts of the compounds described herein that are suitable for pharmaceutical use are salts in which the counterion is pharmaceutically acceptable. However, salts with non-pharmaceutically acceptable counterions may be used, for example, as intermediates in the preparation of the compounds described herein, their pharmaceutically acceptable salts, and physiologically functional derivatives. The term "physiologically functional derivative" refers to a chemical derivative of a compound described herein that has the same physiological function as the compound described herein, for example, that is convertible into a compound described herein in vivo. Esters, amides, and carbamates are examples of physiologically functional derivatives.

[0167] Suitable salt forms of the compounds described herein include those formed with organic or inorganic acids or bases. In particular, suitable salts formed according to the present invention with acids include salts formed with inorganic acids, strong organic carboxylic acids (e.g., unsubstituted or substituted, for example, by halogen, alkanecarboxylic acids having 1 to 4 carbon atoms, or for example, saturated or unsaturated dicarboxylic acids, or for example, hydroxycarboxylic acids, or for example, amino acids) or with organic sulfonic acids (e.g., unsubstituted or substituted, for example, by halogen, (C1-C4) alkyl or arylsulfonic acids). Pharmaceutically acceptable acid addition salts include salts formed with hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, citric acid, tartaric acid, acetic acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, trifluoroacetic acid, succinic acid, perchloric acid, fumaric acid, maleic acid, glycolic acid, lactic acid, salicylic acid, oxaloacetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, isethionic acid, ascorbic acid, malic acid, phthalic acid, aspartic acid and glutamic acid, lysine and arginine. Other acids (e.g., oxalic acid), while not themselves pharmaceutically acceptable, may be used as intermediates in obtaining the compounds described herein and their pharmaceutically acceptable acid addition salts.

[0168] Pharmaceutically acceptable base salts include ammonium salts, alkali metal salts (e.g., potassium and sodium salts), alkaline earth metal salts (e.g., calcium and magnesium salts), and salts with organic bases, such as dicyclohexylamine, N-methyl-D-glucamine, morpholine, thiomorpholine, piperidine, pyrrolidine, mono-, di-, or tri-lower alkylamine (e.g., ethyl, tert-butyl, diethyl, diisopropyl, triethyl, tributyl, or dimethylpropylamine), or mono-, di-, or tri-hydroxy lower alkylamine (e.g., mono-, di-, or triethanolamine). In addition, corresponding inner salts can also be formed.

[0169] Preferred salts of the compounds described herein include acid addition salts, such as salts formed with hydrochloric acid, hydrobromic acid, acetic acid, p-toluenesulfonic acid, tartaric acid, sulfuric acid, succinic acid, phosphoric acid, oxalic acid, nitric acid, methanesulfonic acid, malic acid, maleic acid, and citric acid. More preferably, the salt of the compound described herein is a hydrochloride (i.e., an addition salt formed with hydrochloric acid).

[0170] A compound that is inactive per se but can be converted into an active drug compound after administration to a recipient is called a "prodrug". For example, a prodrug can be converted into an active form with a medical effect in vivo, for example, by hydrolysis in the blood. Pharmaceutically acceptable prodrugs are described in Volume 14 of T. Higuchi and V. Stella, Prodrugs as Novel Delivery Systems, ACS Symposium Series (1976); "Design of Prodrugs", edited by H. Bundgaard, Elsevier, 1985; and Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987.

[0171] Examples of prodrugs include esters, amides, and carbamates.

[0172] The compounds described herein may have suitable groups for conversion to esters, amides, or carbamates. Thus, typical ester and amide groups formed from acidic groups in the compounds described herein include -COOR G 、CONR G 2. SO2OR G or SO2N(R G )2, and the compounds described herein are OH or -NHR G Typical ester and amide and carbamate groups formed by groups include OC(O)R G NR G C(O)R G NR G CO2R G 、OSO2R G and -NR G SO2R G , where R G Selected from the group consisting of: C 1-8 Alkyl, C 2-8 Alkenyl, C 28 Alkynyl, C 3-8 Cycloalkyl and C 3-8 Cycloalkyl C 1-8 Alkyl, halogenated C 18 Alkyl, dihalogenated C 1-8 Alkyl, trihalogenated C 18 Alkyl, phenyl and phenyl C 14 Alkyl; more preferably, R G Selected from the group consisting of: C 1-6 Alkyl, C26 Alkenyl, C 26 Alkynyl, C 3-8 Cycloalkyl and C 3-8 Cycloalkyl C 1-6 alkyl.

[0173] Those skilled in the art of organic chemistry will appreciate that many organic compounds can form complexes with solvents in which they react or precipitate or crystallize. These complexes are referred to as "solvates". For example, a complex formed with water is referred to as a "hydrate". The complex may contain a stoichiometric or non-stoichiometric amount of solvent. Solvates are described in Water-Insoluble Drug Formulation, 2nd edition, R. Lui, CRC Press, page 553, and Byrn et al., Pharm. Res., 12 (7), 1995, 945-954. Before being formulated into a solution, the compounds described herein and their esters, amides, carbamates and / or salts may be in the form of solvates. Solvates of the compounds described herein suitable for use as pharmaceuticals are solvates in which the relevant solvent is a pharmaceutically acceptable solvate. For example, a hydrate is a pharmaceutically acceptable solvate.

[0174] It has been found that the compounds described herein wherein R 5 Selected from the group consisting of: -OC 1-6 Alkyl, formula (V)

[0175]

[0176] Japanese style (Va)

[0177]

[0178] And R b is selected from -N(R e )(R f ) and -OC 1-6 Alkyl groups are easily hydrolyzed in cancer cells to form metabolites that are preferentially sequestered and retained in the cells and have strong alkylating activity.

[0179] Thus, also described herein is a metabolite, wherein the metabolite has a structure according to Formula (I), (Ia) or (Ib), wherein,

[0180] X is C 1-6 alkylene;

[0181] W1, W2, W3 and W4 are each CH, or one of W1, W2, W3 and W4 is N and the rest are CH;

[0182] R1 is selected from the group consisting of: H; C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-4 alkyl; and halogen;

[0183] R 2 is selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-6 alkyl;

[0184] R 3 is a group of formula (II), (III), (IIa) or (IIIa);

[0185] R 4 Selected from the group consisting of: N(R c )(R d ), formula (IV) or formula (IVa);

[0186] When R 4 When it is formula (IV) or (IVa), R 5 is -OH; and when R 4 N(R c )(R d ), R 5 is -OH, formula (V) or formula (Va);

[0187] Each R a Independently selected from the group consisting of: H; -C 16 Alkyl; CH2-phenyl; or -CH2-3 to 12-membered heterocyclic group containing 1, 2, 3 or 4 heteroatoms selected from N, O and S; wherein said -C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of: -OH, -OC 1-6 alkyl, NH2, -NHC(=NH)NH2, -C(O)OH, -C(O)NH2, -SH, -SCH3 and halogen; and the phenyl or heterocyclic group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halogen, -NH2, -OH, -OC 1-6 Alkyl and -NO2; and

[0188] R b for OH;

[0189] or a salt thereof.

[0190] Preferred groups of the above metabolite compounds are as described above for compounds of formula (I), (Ia) and (Ib). For example, in certain embodiments, the metabolite has a structure according to formula (I), (Ia) or (Ib), wherein,

[0191] X is -C 1-2 Alkylene (i.e., -CH2- or -CH2-CH2-);

[0192] R 1 is H;

[0193] R 2 is methyl or phenyl;

[0194] R 3 is formula (IIa) or (IIIa);

[0195] R 4 Selected from the group consisting of: N(R c )(R d ) or formula (IVa);

[0196] When R 4 When it is formula (IVa), R 5 is -OH; and when R 4 N(R c )(R d ), R 5 is -OH or formula (Va);

[0197] Each R a Independently selected from the group consisting of: H; -C 16 Alkyl; CH2-indolyl; CH2-phenyl; or -CH2-5-membered heteroaryl containing 1, 2, 3 or 4 N or S atoms; wherein -C 1-6 The alkyl group is optionally substituted with OH; NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and the phenyl group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halogen; -NH2; -OH; -OC 1-6 alkyl; and -NO2; and

[0198] R b for OH;

[0199] or a salt thereof.

[0200] In certain embodiments, the metabolite has a structure according to Formula (I), (Ia), or (Ib), wherein,

[0201] X is -C 1-2 Alkylene (i.e., -CH2- or -CH2-CH2-);

[0202] R 1 is H;

[0203] R 2 is methyl or phenyl;

[0204] R 3 is formula (IIa) or (IIIa);

[0205] R 4 is NH2 or formula (IVa);

[0206] R 5 is -OH;

[0207] Each R a Independently selected from the group consisting of: H; -C 16 Alkyl; CH2-indolyl; CH2-phenyl; or -CH2-5-membered heteroaryl containing 1, 2, 3 or 4 N or S atoms; wherein C 1-6 The alkyl group is optionally substituted with OH; NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and the phenyl group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halogen; -NH2; -OH; -OC 1-6 alkyl; and -NO2. Preferably, each R a independently selected from the group consisting of isopropyl, isobutyl, and -CH2-phenyl optionally substituted with 1 F (e.g., 2-fluorobenzyl, 3-fluorobenzyl, or 4-fluorobenzyl);

[0208] or a salt thereof.

[0209] In certain preferred embodiments, the metabolite is:

[0210] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-methyl-butanoic acid (Example Compound 19); and

[0211] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (Example Compound 20);

[0212] (2S)-2-[[(2S)-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]-2-(methylamino)butanoyl]amino]-4-methyl-pentanoic acid (Example Compound 33)

[0213] (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid (Example Compound 37)

[0214] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-(4-fluorophenyl)propanoic acid (Example Compound 39)

[0215] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-phenyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (Example Compound 40)

[0216] 2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-3-methyl-butanoic acid (Example Compound 42)

[0217] (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propanoic acid (Example Compound 43)

[0218] (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid (Exemplary Compound 44); and

[0219] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloro-1,1,2,2-tetradeuterium-ethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (Example Compound 51),

[0220] or a salt thereof.

[0221] It has been found that the compounds described herein are readily further hydrolyzed within cancer cells to form higher order metabolites that are preferentially sequestered and retained within the cell and possess potent alkylating activity. Thus, also described herein is a metabolite having a structure according to Formula (I), (Ia), or (Ib), wherein X is C 1-6 alkylene;

[0222] W1, W2, W3 and W4 are each CH, or one of W1, W2, W3 and W4 is N and the rest are CH;

[0223] R 1is selected from the group consisting of: H; -C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-4 Alkyl; and halogen (such as F or Cl);

[0224] R 2 is selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen (such as F or Cl); and -C 1-6 alkyl; and

[0225] R 3 Formula (VIa)

[0226]

[0227] where R c and R d Each independently selected from the group consisting of: H, -C 1-6 Alkyl, -C(O)C 1-6 Alkyl and -CH2-phenyl, wherein the alkyl or the phenyl is optionally substituted by 1, 2 or 3 groups selected from halogen (preferably, R c and R d are H); and

[0228] R g Selected from the group consisting of: H and -C 1-6 Alkyl, wherein the alkyl is optionally substituted by 1, 2 or 3 groups selected from halogen (preferably, R g for H).

[0229] For the avoidance of doubt, in formulas (VI) and (VIa) represents the point of attachment of Formula (VI) or (VIa) to Formula (I), (Ia) or (Ib).

[0230] In certain embodiments, the metabolite has a structure according to Formula (I), (Ia), or (Ib), wherein,

[0231] X is C 1-6 alkylene;

[0232] R 1 Selected from the group consisting of: H; -C 1-4 Alkyl; and halogen (such as F or Cl);

[0233] R 2 is selected from the group consisting of: H; phenyl substituted by 1, 2 or 3 halogens (such as F or Cl); and -C 1-6Alkyl (e.g., selected from the group consisting of: phenyl substituted with 1, 2, or 3 halogens (e.g., F or Cl); and -C 1-6 alkyl));

[0234] R 3 is formula (VIa); and

[0235] R c and R d All are H;

[0236] or a salt thereof.

[0237] In certain other embodiments, the metabolite has a structure according to Formula (I), (Ia), or (Ib), wherein,

[0238] X is C1 alkylene (i.e., -CH2-) or C2 alkylene (i.e., -CH2-CH2-);

[0239] R 1 Selected from the group consisting of: H; -C 1-4 Alkyl; and halogen (such as F or Cl);

[0240] R 2 is selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 halogens (such as F or Cl); and -C 1-6 alkyl; and

[0241] R 3 is formula (VIa); and

[0242] R c and R d All are H;

[0243] or a salt thereof.

[0244] Preferably, the compound is a compound of formula (Ia) or (Ib).

[0245] Thus, also described herein is a compound having a structure according to Formula (I), (Ia) or (Ib), wherein,

[0246] X is C 2-6 alkylene;

[0247] W1, W2, W3 and W4 are each CH, or one of W1, W2, W3 and W4 is N and the rest are CH;

[0248] R 1 is selected from the group consisting of: H; -C optionally substituted by 1, 2 or 3 groups independently selected from halogen1-4 Alkyl; and halogen (such as F or Cl);

[0249] R 2 is selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen (such as F or Cl); and -C 1-6 alkyl; and

[0250] R 3 Formula (VI)

[0251]

[0252] where R c and R d Each independently selected from the group consisting of: H, -C 1-6 Alkyl, -C(O)C 1-6 Alkyl and -CH2-phenyl, wherein the alkyl or the phenyl is optionally substituted by 1, 2 or 3 groups selected from halogen (e.g., R c and R d are H); and

[0253] R g Selected from the group consisting of: H and -C 1-6 Alkyl, wherein the alkyl is optionally substituted with 1, 2 or 3 groups selected from halogen (e.g., R g for H).

[0254] Negatively charged cyclodextrin

[0255] Negatively charged cyclodextrins are cyclodextrins or cyclodextrin derivatives that carry one or more negative charges.

[0256] For example, the negatively charged cyclodextrin or its derivative has 4 to 10 negative charges. Preferably, the negatively charged cyclodextrin or its derivative has 5 to 8 negative charges.

[0257] Preferably, the cyclodextrin or its derivative carries one or more negative charges at a pH above 2 (eg, pH 2 to pH 8 or pH 3 to pH 5.5).

[0258] Preferably, the negative charge is located in a substituent of the cyclodextrin, for example in a carboxylic acid group or a sulfoalkyl group, preferably in a sulfoalkyl group, such as a sulfobutyl group.

[0259] The average number of sulfoalkyl groups (such as sulfobutyl groups) per cyclodextrin ring, expressed as an average degree of substitution, is 4 to 10. Preferably, the average number of sulfoalkyl groups (such as sulfobutyl groups) per cyclodextrin ring, expressed as an average degree of substitution, is 5 to 8.

[0260] The negatively charged cyclodextrin or its derivative can be of any size, such as negatively charged α-cyclodextrin or its derivative, negatively charged β-cyclodextrin or its derivative, or negatively charged γ-cyclodextrin or its derivative. Preferably, the negatively charged cyclodextrin or its derivative is negatively charged β-cyclodextrin or its derivative.

[0261] A preferred negatively charged cyclodextrin is a sulfoalkyl ether β-cyclodextrin, wherein the number of sulfoalkyl groups per cyclodextrin ring, expressed as an average degree of substitution, is from 4 to 10. More preferably, the negatively charged cyclodextrin is a sulfobutyl ether β-cyclodextrin, wherein the average number of sulfobutyl groups per cyclodextrin ring, expressed as an average degree of substitution, is from 5 to 8. The structural formula of such a sulfobutyl ether β-cyclodextrin is shown below, where m is an exemplary value for the degree of substitution.

[0262]

[0263] A particularly preferred negatively charged cyclodextrin is sodium sulfobutyl ether beta-cyclodextrin (also known as "Dexolve" or "Captisol"), as described in Handbook of Pharmaceutical excipients. 8th edition London: Pharmaceutical Press (2017), pages 112-117.

[0264] pharmaceutical preparations

[0265] It has been found that formulations comprising a compound of formula (I) as described above and a negatively charged cyclodextrin are particularly stable. Accordingly, the present invention provides a pharmaceutical formulation comprising a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin and a compound according to formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate.

[0266] Preferably, the pharmaceutical formulation of the present invention comprises a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin having 4 to 10 negative charges, more preferably 5 to 8 negative charges.

[0267] Preferably, the pharmaceutical formulation of the present invention comprises a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, which carries one or more negative charges at a pH above 2, for example, pH 2 to pH 8 or pH 3 to pH 5.5.

[0268] Preferably, the pharmaceutical formulation of the present invention comprises a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, such as a carboxylic acid-derived cyclodextrin or a sulfoalkyl ether-derived cyclodextrin. More preferably, the pharmaceutical formulation of the present invention comprises a sulfoalkyl ether-derived cyclodextrin, such as a sulfobutyl ether-derived cyclodextrin.

[0269] When the pharmaceutical formulation of the present invention comprises a sulfoalkyl ether-derivatized cyclodextrin (e.g., a sulfobutyl ether-derivatized cyclodextrin), the average number of sulfoalkyl groups (e.g., sulfobutyl groups) per cyclodextrin ring is preferably 4 to 10, more preferably 5 to 8, expressed as the average degree of substitution.

[0270] Preferably, the pharmaceutical formulation of the present invention comprises negatively charged α-cyclodextrin or a pharmaceutically acceptable derivative of negatively charged α-cyclodextrin, negatively charged β-cyclodextrin or a pharmaceutically acceptable derivative of β-cyclodextrin, or negatively charged γ-cyclodextrin or a pharmaceutically acceptable derivative of γ-cyclodextrin. More preferably, the pharmaceutical formulation of the present invention comprises negatively charged β-cyclodextrin or a pharmaceutically acceptable derivative of β-cyclodextrin.

[0271] Preferably, the pharmaceutical formulation of the present invention comprises a sulfoalkyl ether β-cyclodextrin, wherein the number of sulfoalkyl groups per cyclodextrin ring, expressed as an average degree of substitution, is between 4 and 10.

[0272] More preferably, the pharmaceutical formulation of the present invention comprises sulfobutyl ether β-cyclodextrin, wherein the average number of sulfobutyl groups per cyclodextrin ring, expressed as the average degree of substitution, is from 5 to 8.

[0273] Even more preferably, the pharmaceutical formulation of the present invention comprises sulfobutyl ether β-cyclodextrin having the structure shown below (wherein m is an exemplary value for the degree of substitution):

[0274]

[0275] Most preferably, the pharmaceutical formulation of the present invention comprises sodium sulfobutyl ether beta-cyclodextrin. The pharmaceutical formulation of the present invention comprises a compound according to formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate

[0276]

[0277] in,

[0278] X is C 1-6 alkylene;

[0279] W1, W2, W3 and W4 are each CH, or one of W1, W2, W3 and W4 is N and the rest are CH;

[0280] R 1 is selected from the group consisting of: H; C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-4 alkyl; and halogen;

[0281] R 2is selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-6 alkyl; and

[0282] R 3 is a group according to formula (II):

[0283]

[0284] or a group according to formula (III):

[0285]

[0286] in,

[0287] R 4 Selected from the group consisting of: N(R c )(R d ) and formula (IV):

[0288] and

[0289] When R 4 When it is formula (IV), R 5 R b ; and when R 4 N(R c )(R d ), R 5 Selected from the group consisting of: R b and formula (V):

[0290]

[0291] Among them, each R a Independently selected from the group consisting of: H; C 16 Alkyl; CH2-phenyl; or -CH2-3 to 12-membered heterocyclic group containing 1, 2, 3 or 4 heteroatoms selected from N, O and S; wherein said C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of: -OH, -OC 1-6 alkyl, NH2, -NHC(=NH)NH2, -C(O)OH, -C(O)NH2, -SH, -SCH3 and halogen; and the phenyl or heterocyclic group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halogen, -NH2, -OH, -OC 1-6 Alkyl and -NO2;

[0292] R b Selected from the group consisting of: -OH; -N(Re )(R f ); and -OC optionally substituted by one or more groups selected from the following 1-6 Alkyl: halogen, -OH, -CN, -N(R e )(R f ),-C 6-10 aryl or a 3- to 12-membered heterocyclic ring comprising one or more O, N or S atoms and optionally substituted by 1, 2 or 3 halogens, and / or wherein the alkyl is optionally interrupted by 1, 2 or 3 O, N or S atoms;

[0293] R c and R d Each independently selected from the group consisting of: H; -C 16 Alkyl; C(O)C 16 alkyl; and -CH2-phenyl, wherein the alkyl or the phenyl is optionally substituted with 1, 2 or 3 groups selected from halogen; and

[0294] R e and R f Each independently selected from the group consisting of: H and -C 16 Alkyl, wherein the alkyl is optionally substituted with 1, 2 or 3 groups selected from halogen; or R e and R f Together with the nitrogen atom to which they are attached they form a 4-, 5- or 6-membered heterocyclic ring which is optionally substituted by 1, 2 or 3 groups selected from halogen.

[0295] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, which has structure (Ia):

[0296]

[0297] For example, structure (Ib):

[0298]

[0299] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I), (Ia) or (Ib) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein R 1 For H.

[0300] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I), (Ia) or (Ib) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein R 2is selected from the group consisting of: phenyl optionally substituted by 1, 2 or 3 halogens; and -C 1-6 Alkyl; for example -C optionally substituted by 1, 2 or 3 halogen 1-4 Alkyl groups, such as methyl.

[0301] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I), (Ia) or (Ib) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein X is -CH2- or -CH2-CH2-.

[0302] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I), (Ia) or (Ib) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein R 3 is a group according to formula (IIa):

[0303]

[0304] or a group according to formula (IIIa):

[0305]

[0306] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I), (Ia) or (Ib) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein R 4 Selected from the group consisting of: N(R c )(R d ) and formula (IVa):

[0307]

[0308] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I), (Ia) or (Ib) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein when R 4 When it is formula (IV) or (IVa), R 5 R b ; and when R 4 N(R c )(R d ), R 5 Selected from the group consisting of: R b and formula (Va), wherein formula (Va) has the following structure,

[0309]

[0310] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I), (Ia) or (Ib) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein R 4 is NH2.

[0311] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I), (Ia) or (Ib) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein each R a Independently selected from the group consisting of: C 16 Alkyl and CH2-phenyl; wherein said C 1-6 The alkyl group is optionally substituted with OH; NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen (e.g., F or Cl); and the phenyl group is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of halogen (e.g., F or Cl); -NH2; -OH; -OC 1-6 Alkyl; and -NO2.

[0312] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I), (Ia) or (Ib) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein each R a Selected from the group consisting of: -C 16 alkyl and CH2-phenyl; wherein the phenyl is optionally substituted with 1, 2 or 3 halogen substituents.

[0313] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I), (Ia) or (Ib) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein when R 4 When it is formula (IV) or (IVa), R 5 For-OC 1-6 Alkyl; and when R 4 When it is NH2, R 5 For-OC 1-6 Alkyl or formula (V) or (Va).

[0314] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I), (Ia) or (Ib) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein when R b For-OC 1-6 When alkyl, it is selected from the group consisting of methoxy, ethoxy and isopropoxy.

[0315] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, which is selected from the group consisting of:

[0316] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 1);

[0317] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-(4-fluorophenyl)propanoic acid ethyl ester (Example Compound 2);

[0318] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-methyl-butyric acid ethyl ester (Example Compound 3);

[0319] (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 4);

[0320] (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-3-(4-fluorophenyl)propionic acid ethyl ester (Example Compound 5);

[0321] (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-3-methyl-butyric acid ethyl ester (Example Compound 6);

[0322] (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 7);

[0323] (2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propionyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Exemplary Compound 8);

[0324] (2S)-2-[[(2S)-2-amino-3-methyl-butyryl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 9);

[0325] (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionic acid ethyl ester (Example Compound 10);

[0326] (2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propionyl]amino]-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionic acid ethyl ester (Example Compound 11);

[0327] (2S)-2-[[(2S)-2-amino-3-methyl-butyryl]amino]-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionic acid ethyl ester (Example Compound 12);

[0328] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-phenyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 13);

[0329] (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid methyl ester (Example Compound 14);

[0330] (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid isopropyl ester (Exemplary Compound 15);

[0331] (2S)-2-[[(2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propionyl]amino]-4-methyl-pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 16);

[0332] (2S)-2-[[(2R)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propionyl]amino]-4-methyl-pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 21)

[0333] (2S)-2-[[(2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoyl]amino]-3-(4-fluorophenyl)propionic acid ethyl ester (Example Compound 22)

[0334] (2S)-2-[[(2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 23)

[0335] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid 2-morpholinoethyl ester (Example Compound 24)

[0336] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid 2-isopropoxyethyl ester (Example Compound 25)

[0337] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-valeric acid isopropyl ester (Example Compound 26)

[0338] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid methyl ester (Example Compound 27)

[0339] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid 3-(dimethylamino)propyl ester (Example Compound 28)

[0340] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (2-methoxy-1-methyl-ethyl ester) (Example Compound 29)

[0341] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-N,N,4-trimethyl-pentanamide (Example Compound 30)

[0342] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-N,4-dimethyl-pentanamide (Example Compound 31)

[0343] (2S)-2-[[(2S)-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]-2-(methylamino)butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 32)

[0344] (2S)-2-[[(2S)-2-acetylamino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-(4-fluorophenyl)propionic acid ethyl ester (Example Compound 34)

[0345] (2S)-2-[[(2S)-2-Acetylamino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 35)

[0346] (2S)-2-[[(2S)-2-acetylamino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-3-(4-fluorophenyl)propionic acid ethyl ester (Example Compound 36)

[0347] (2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 38)

[0348] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 45)

[0349] (2S)-2-[[(2R)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 46)

[0350] (2R)-2-[[(2R)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 47)

[0351] (2S)-2-[[(2S)-2-amino-4-[6-[bis(2-chloroethyl)amino]-3-methyl-imidazo[4,5-b]pyridin-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Exemplary Compound 48); and

[0352] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloro-1,1,2,2-tetradeuterium-ethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 50)

[0353] or a pharmaceutically acceptable salt, amide or carbamate thereof, including salts of such amides or carbamates.

[0354] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, which is selected from the group consisting of:

[0355] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-methyl-butanoic acid (Example Compound 19); and

[0356] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (Example Compound 20);

[0357] (2S)-2-[[(2S)-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]-2-(methylamino)butanoyl]amino]-4-methyl-pentanoic acid (Example Compound 33)

[0358] (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid (Example Compound 37)

[0359] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-(4-fluorophenyl)propanoic acid (Example Compound 39)

[0360] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-phenyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (Example Compound 40)

[0361] 2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-3-methyl-butanoic acid (Example Compound 42)

[0362] (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propanoic acid (Example Compound 43)

[0363] (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid (Exemplary Compound 44); and

[0364] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloro-1,1,2,2-tetradeuterium-ethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (Example Compound 51)

[0365] or a salt thereof.

[0366] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein:

[0367] X is C 1-6 alkylene;

[0368] W1, W2, W3 and W4 are each CH, or one of W1, W2, W3 and W4 is N and the rest are CH;

[0369] R 1 is selected from the group consisting of: H; C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-4 alkyl; and halogen;

[0370] R 2 is selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-6 alkyl; and

[0371] R 3 Formula (VIa):

[0372]

[0373] where R c and R dEach independently selected from the group consisting of: H, C 16 Alkyl, C(O)C 16 alkyl and -CH2-phenyl, wherein the alkyl or the phenyl is optionally substituted with 1, 2 or 3 groups selected from halogen; and

[0374] R g Selected from the group consisting of: H and C 16 Alkyl, wherein the alkyl is optionally substituted with 1, 2 or 3 groups selected from halogen.

[0375] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (Ia) or (Ib) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate,

[0376]

[0377] Where X is C 1-6 alkylene;

[0378] W1, W2, W3 and W4 are each CH, or one of W1, W2, W3 and W4 is N and the rest are CH;

[0379] R 1 is selected from the group consisting of: H; C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-4 alkyl; and halogen;

[0380] R 2 is selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-6 alkyl; and

[0381] R 3 Formula (VIa):

[0382]

[0383] where R c and R d Each independently selected from the group consisting of: H, C 16 Alkyl, C(O)C 16 Alkyl and -CH2-phenyl, wherein the alkyl or the phenyl is optionally substituted with 1, 2 or 3 groups selected from halogen;

[0384] R g Selected from the group consisting of: H and C 16 Alkyl, wherein the alkyl is optionally substituted with 1, 2 or 3 groups selected from halogen.

[0385] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein:

[0386] X is C 1-6 alkylene;

[0387] W1, W2, W3 and W4 are each CH, or one of W1, W2, W3 and W4 is N and the rest are CH;

[0388] R 1 is H;

[0389] R 2 is selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-6 alkyl; and

[0390] R 3 Formula (VIa):

[0391]

[0392] where R c and R d are H; and

[0393] R g For H.

[0394] Preferably, the pharmaceutical preparation of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, wherein X is C 1-6 alkylene;

[0395] W1, W2, W3 and W4 are each CH, or one of W1, W2, W3 and W4 is N and the rest are CH;

[0396] R 1 is selected from the group consisting of: H; C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-4 alkyl; and halogen;

[0397] R 2 is selected from the group consisting of: phenyl optionally substituted by 1, 2 or 3 halogens; and C 1-6 Alkyl (e.g., optionally substituted with 1, 2, or 3 halogens) 1-4 alkyl, such as methyl); and

[0398] R3 Formula (VIa):

[0399]

[0400]

[0401] where R c and R d are H; and

[0402] R g For H.

[0403] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate,

[0404] wherein X is -CH2- or -CH2-CH2-;

[0405] W1, W2, W3 and W4 are each CH, or one of W1, W2, W3 and W4 is N and the rest are CH;

[0406] R 1 is selected from the group consisting of: H; C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-4 alkyl; and halogen;

[0407] R 2 is selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-6 alkyl; and

[0408] R 3 Formula (VIa):

[0409]

[0410] where R c and R d are H; and

[0411] R g For H.

[0412] Preferably, the pharmaceutical formulation of the present invention comprises a compound according to formula (I) or a salt or solvate thereof, wherein:

[0413] X is C 1-6 alkylene;

[0414] W1, W2, W3 and W4 are each CH, or one of W1, W2, W3 and W4 is N and the rest are CH;

[0415] R 1 is selected from the group consisting of: H; -C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-4 alkyl; and halogen;

[0416] R 2 is selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and -C 1-6 alkyl;

[0417] R 3 Formula (VI)

[0418]

[0419] where R c and R d Each independently selected from the group consisting of: H, C 16 Alkyl, -C(O)C 16 alkyl and -CH2-phenyl, wherein the alkyl or the phenyl is optionally substituted with 1, 2 or 3 groups selected from halogen; and

[0420] R g Selected from the group consisting of: H and C 16 Alkyl, wherein the alkyl is optionally substituted with 1, 2 or 3 groups selected from halogen.

[0421] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, which is selected from the group consisting of:

[0422] (2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propanoic acid (Example Compound 17);

[0423] (S)-2-amino-4-(5-(bis(2-chloroethyl)amino)-1-methyl-1H-benzo[d]imidazol-2-yl)butanoic acid (Example Compound 18);

[0424] (2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 38)

[0425] (2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-phenyl-benzimidazol-2-yl]butanoic acid (Exemplary Compound 41); and

[0426] (2S)-2-amino-4-[5-[bis(2-chloro-1,1,2,2-tetradeuterium-ethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid (Example Compound 49);

[0427] or a salt thereof.

[0428] Preferably, the pharmaceutical formulation of the present invention comprises (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Exemplary Compound 1).

[0429] Preferably, the compound of formula (I) or its ester, amide or carbamate is present in the pharmaceutical preparation of the present invention in the form of a pharmaceutically acceptable salt, more preferably, the compound of formula (I) or its ester, amide or carbamate is present in the form of a hydrochloride.

[0430] Preferably, the pharmaceutical preparation of the present invention comprises sodium sulfobutyl ether beta-cyclodextrin and (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Exemplary Compound 1).

[0431] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such ester, amide or carbamate (excluding the mass of any counter ion), and the amount of each administration is about 1 mg to 150 mg. For example, 1 mg, 2 mg, 3 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg or 150 mg. Alternatively, the compound of the present invention or its salt and / or solvate (excluding the mass of any counter ion or solvent) can be administered in a single high dose. A single high dose can be about 150 mg to 800 mg. For example, 150 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg or 800 mg.

[0432] Preferably, the pharmaceutical formulations of the present invention comprise a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the weight ratio (w / w) of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of a negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate is from about 1:1 to about 100: 1. Typically, the pharmaceutical formulations comprise a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the weight ratio (w / w) of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of a negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate is from about 4:1 to about 80:1, from about 3:1 to about 60:1, from about 2:1 to about 50:1, from about 4:1 to about 30:1, or from about 5:1 to about 20:1.

[0433] Preferably, the pharmaceutical formulation of the present invention comprises a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the weight ratio (w / w) of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate is from about 5: 1 to about 20: 1. In an exemplary embodiment, the pharmaceutical formulation of the present invention may comprise a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the weight ratio (w / w) of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate is at least about 5: 1. The pharmaceutical formulation of the present invention may comprise a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the weight ratio (w / w) of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate is about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1 or about 25:1.

[0434] Preferably, the pharmaceutical formulation of the present invention comprises a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the weight ratio (w / w) of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate is about 5:1, about 10:1 or about 20:1.

[0435] Preferably, the pharmaceutical preparation of the present invention comprises sodium sulfobutyl ether beta-cyclodextrin and (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butyryl]amino]-4-methyl-pentanoic acid ethyl ester (exemplified compound 1) in a weight ratio of 5:1 to 20:1.

[0436] Preferably, the pharmaceutical preparation of the present invention comprises sodium sulfobutyl ether beta-cyclodextrin and (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butyryl]amino]-4-methyl-pentanoic acid ethyl ester (exemplified compound 1) in a weight ratio of 10:1.

[0437] In exemplary embodiments, wherein the pharmaceutical formulation of the present invention comprises a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the weight ratio (w / w) of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate is from about 5:1 to about 20:1, the pharmaceutical formulation may comprise, for example, 1 mg to 50 mg of the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate (excluding the mass of any counterion) and about 5 mg to about 1000 mg of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin. For example, a pharmaceutical formulation containing 20 mg of a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate (excluding the mass of any counterion) can contain from about 100 mg to about 400 mg of a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin (e.g., from about 200 mg to about 300 mg of a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin).

[0438] Additional excipients may be included in the pharmaceutical formulations of the present invention. Other excipients may also be included in the pharmaceutical formulations, for example, to increase the dissolution rate of the compound of formula (I) or its pharmaceutically acceptable salt, ester, amide or carbamate, or salt of such ester, amide or carbamate in aqueous solution and / or to increase the stability of the compound of formula (I) or its pharmaceutically acceptable salt, ester, amide or carbamate, or salt of such ester, amide or carbamate in its lyophilized form.

[0439] The one or more additional pharmaceutically acceptable excipients may be selected from the group consisting of polysorbate 80, propylene glycol, sucrose, trehalose dihydrate, MgCl , CaCl , citric acid, citrates (e.g., trisodium citrate dihydrate), lactic acid and lactates, tartaric acid and tartrates, phosphoric acid and phosphates, glycine, maleic acid, succinic acid and succinates, aspartic acid, benzoic acid and benzoate. Preferably, the other pharmaceutically acceptable excipients are selected from citric acid and citrates (e.g., trisodium citrate dihydrate).

[0440] Typically, when the pharmaceutical formulations of the present invention comprise one or more other excipients in addition to the negatively charged cyclodextrin or a pharmaceutically acceptable derivative of the negatively charged cyclodextrin, the one or more other excipients are each independently present in the formulation at a weight ratio (w / w) of excipient to compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, of from about 0.1:1 to about 100:1, for example, from about 1:1 to about 50:1, for example, from about 1:1 to about 30:1, from about 1:1 to about 20:1, or from about 1:1 to about 10:1, for example, from about 1.0:1 to about 7.5:1, from about 1.0:1 to about 5:1, from about 1.0:1 to about 2.0:1.

[0441] In certain embodiments, the pharmaceutical formulations of the present invention are lyophilized powders. It has been found that negatively charged cyclodextrins or pharmaceutically acceptable derivatives of negatively charged cyclodextrins can be included in lyophilized formulations of compounds of formula (I) or pharmaceutically acceptable salts, esters, amides, or carbamates thereof, or salts of such esters, amides, or carbamates. Reconstitution of the lyophilized powder provides aqueous solutions in which compounds of formula (I) or pharmaceutically acceptable salts, esters, amides, or carbamates thereof, or salts of such esters, amides, or carbamates, are particularly stable.

[0442] The pharmaceutical preparation of the present invention, when in the form of a lyophilized powder, can be essentially composed of a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such ester, amide or carbamate and a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin. Alternatively, the pharmaceutical preparation of the present invention, when in the form of a lyophilized powder, in addition to a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, also comprises one or more other excipients. The one or more other excipients can be selected from the group consisting of: sucrose, polysorbate 80, propylene glycol, trehalose dihydrate, MgCl2, CaCl2, citric acid, citrate (e.g., trisodium citrate dihydrate), lactic acid and lactate, tartaric acid and tartrate, phosphoric acid and phosphate, glycine, maleic acid, succinic acid and succinate, aspartic acid, benzoic acid and benzoate. Preferably, the other excipients are selected from citric acid or citrate (e.g., trisodium citrate dihydrate).

[0443] Preferably, the pharmaceutical preparation of the present invention comprises sodium sulfobutyl ether beta-cyclodextrin, (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butyryl]amino]-4-methyl-pentanoic acid ethyl ester (exemplified compound 1), citric acid and trisodium citrate dihydrate.

[0444] Preferably, the pharmaceutical preparation of the present invention comprises sodium sulfobutyl ether beta-cyclodextrin, (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butyryl]amino]-4-methyl-pentanoic acid ethyl ester (Exemplary Compound 1), citric acid and trisodium citrate dihydrate, wherein sodium sulfobutyl ether beta-cyclodextrin and (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butyryl]amino]-4-methyl-pentanoic acid ethyl ester (Exemplary Compound 1) are present in a weight ratio of 5:1 to 20:1. The other excipients, citric acid and citrate salts (e.g., trisodium citrate dihydrate), and the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide, or carbamate thereof, or a salt of such an ester, amide, or carbamate (e.g., exemplary compound 1), are present in a weight ratio, for example, of about 0.1:1 to about 100:1. For example, the ratio can be from about 1:1 to about 50:1, for example, from about 1:1 to about 30:1, from about 1:1 to about 20:1, or from about 1:1 to about 10:1, for example, from about 1.0:1 to about 7.5:1, from about 1.0:1 to about 5:1, or from about 1.0:1 to about 2.0:1.

[0445] Preferably, the pharmaceutical preparation of the present invention comprises sodium sulfobutyl ether beta-cyclodextrin, (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butyryl]amino]-4-methyl-pentanoic acid ethyl ester (Exemplary Compound 1), citric acid and trisodium citrate dihydrate, wherein sodium sulfobutyl ether beta-cyclodextrin and (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butyryl]amino]-4-methyl-pentanoic acid ethyl ester (Exemplary Compound 1) are present in a weight ratio of 10:1. The other excipients, citric acid and citrate salts (e.g., trisodium citrate dihydrate), and the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide, or carbamate thereof, or a salt of such an ester, amide, or carbamate (e.g., exemplary compound 1), are present in a weight ratio, for example, of about 0.1:1 to about 100:1. For example, the ratio can be from about 1:1 to about 50:1, for example, from about 1:1 to about 30:1, from about 1:1 to about 20:1, or from about 1:1 to about 10:1, for example, from about 1.0:1 to about 7.5:1, from about 1.0:1 to about 5:1, or from about 1.0:1 to about 2.0:1.

[0446] Preferably, in embodiments where the pharmaceutical formulation is a lyophilized powder, the pharmaceutical formulation is free or substantially free of any organic solvent. By "free" of any organic solvent, it is meant that the lyophilized powder pharmaceutical formulation does not contain any measurable amount of organic solvent. By "substantially free" of any organic solvent, it is meant that the lyophilized powder pharmaceutical formulation contains only trace amounts of organic solvent, e.g., less than about 0.1% w / v of the total amount of organic solvent.

[0447] In certain embodiments, the pharmaceutical preparations of the present invention are liquid pharmaceutical formulations. That is, the formulations are in liquid form when stored at room temperature.

[0448] The present invention also provides a composition comprising a pharmaceutical formulation of the present invention and a physiologically acceptable aqueous solvent or diluent. Preferably, the composition of the present invention comprises a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, and a physiologically acceptable aqueous solvent.

[0449] Preferably, the physiologically acceptable aqueous solvent used in the composition of the present invention is a glucose solution, a saline solution or a mixture thereof.

[0450] The compositions of the present invention may be suitable for parenteral administration to a subject. Injections and infusions may be prepared by diluting the lyophilized powder or liquid pharmaceutical formulation of the present invention with one or more physiologically acceptable solvents or diluents. Exemplary solvents or diluents include mannitol, glucose, 1,3-butylene glycol, water, Ringer's solution, isotonic sodium chloride solution or other suitable dispersants or wetting agents and suspending agents, including synthetic mono- or diglycerides and fatty acids, including oleic acid or Cremaphor.

[0451] In certain embodiments, the pharmaceutical formulations of the present invention are administered to a subject after dilution, for example, with a physiologically acceptable aqueous solvent or diluent, such as saline solution, glucose solution, or a mixture thereof. Any such solvent or diluent may optionally be buffered.

[0452] Preferably, the physiologically acceptable solvent is an aqueous solution, such as a glucose solution (eg a 5% glucose solution (w / v in water)), a saline solution (eg a 0.9% sodium chloride solution (w / v in water)), or a mixture thereof.

[0453] The pharmaceutical formulations of the present invention can be diluted with a physiologically acceptable solvent, for example, to a concentration of the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate (excluding the mass of any counterion) of about 0.001 mg / mL to about 3 mg / mL (e.g., 0.01 mg / mL to 2 mg / mL, 0.05 mg / mL to 1.4 mg / mL, 0.05 mg / mL to 1.2 mg / mL, 0.05 mg / mL to 1 mg / mL, 0.05 mg / mL to 0.9 mg / mL, 0.05 mg / mL to 0.8 mg / mL, 0.1 mg / mL to 0.4 mg / mL, or 0.1 mg / mL to 0.3 mg / mL), and then administered to a subject.

[0454] The composition of the present invention suitable for parenteral administration is preferably suitable for administration by infusion or injection, and is particularly suitable for administration by intravenous infusion.

[0455] In certain embodiments, the compound of formula (I) or its pharmaceutically acceptable salt, ester, amide or carbamate, or such ester, amide or carbamate salt can be prepared by diluting a lyophilized powder of a compound of formula (I) or its pharmaceutically acceptable salt, ester, amide or carbamate, or such ester, amide or carbamate salt that does not contain a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin with one or more physiologically acceptable solvents or diluents containing a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, For example, the compound of formula (I) or its pharmaceutically acceptable salt, ester, amide or carbamate, or such ester, amide or carbamate salt can be a lyophilized powder comprising (or consisting essentially of) a compound of formula (I) or its pharmaceutically acceptable salt, ester, amide or carbamate, or such ester, amide or carbamate salt.

[0456] In embodiments where one or more physiologically acceptable solvents or diluents contain a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, the negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin can be present at a concentration of about 0.1 mg / mL to 10 mg / mL. For example, the negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin can be present at a concentration of about 0.1 mg / mL to 3 mg / mL, for example, about 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, 0.9 mg / mL, 1 mg / mL, 1.5 mg / mL, 2 mg / mL, 2.5 mg / mL, 3 mg / mL, 4 mg / mL or 5 mg / mL.

[0457] Alternatively or additionally, the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin may be added directly to the injection or infusion solution after reconstitution and / or dilution of the compound of formula (I) or its pharmaceutically acceptable salt, ester, amide or carbamate, or the salt of such ester, amide or carbamate with a suitable physiologically acceptable solvent or diluent; or the compound of formula (I) or its pharmaceutically acceptable salt, ester, amide or carbamate, or the salt of such ester, amide or carbamate may be added directly to the injection or infusion solution after reconstitution and / or dilution of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin with a suitable physiologically acceptable solvent or diluent. Therefore, the present invention also provides an injection or infusion solution comprising a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, one or more physiologically acceptable solvents or diluents, and optionally one or more other excipients.

[0458] In embodiments where the negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin is added directly to the injection or infusion solution after reconstitution and / or dilution of the compound of formula (I) or its pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such ester, amide or carbamate with a suitable physiologically acceptable solvent or diluent; or where the negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin is added directly to the injection or infusion solution after reconstitution and / or dilution of the negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin with a suitable physiologically acceptable solvent or diluent, the compound of formula (I) or its pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such ester, amide or carbamate can be in the form of a pharmaceutical preparation, for example, a lyophilized powder comprising the compound of formula (I) or its pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such ester, amide or carbamate.

[0459] In an embodiment of providing an injection or infusion solution comprising a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, one or more physiologically acceptable solvents or diluents, and optionally one or more other excipients, the injection or infusion solution preferably comprises a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of a negatively charged cyclodextrin is in admixture with a compound of formula (I). I) a compound or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, in a weight ratio (w / w) of about 1:1 to about 100:1, for example comprising a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the weight ratio (w / w) of the negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, is about 4:1 to about 80:1 or about 2:1 to about 50:1. In one embodiment, the injection or infusion solution comprises a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the weight ratio (w / w) of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate is from about 4:1 to about 80:1 or from about 5:1 to about 20:1. For example, the injection or infusion solution comprises a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the weight ratio (w / w) of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate is about 5:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1 or about 25:1. In a preferred embodiment, the weight ratio (w / w) of the negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate is about 5:1, about 10:1 or about 20:1.

[0460] Additional therapeutic agents

[0461] Although the compound of formula ( ) or its salt may be present in the pharmaceutical preparation of the present invention as the sole active ingredient, the pharmaceutical preparation of the present invention may also contain one or more additional therapeutic agents. Such agents are known in the art. Examples of other therapeutic agents used in the present invention include steroids (prednisone, prednisolone and dexamethasone), IMiDs (thalidomide, lenalidomide and pomalidomide), PIs (bortezomib, carfilzomib and ixazomib), histone deacetylase (HDAC) inhibitors (panobinostat), conventional chemotherapy (alkylating agents (e.g., melflufen, melphalan, cyclophosphamide, bendamustine), doxorubicin), checkpoint inhibitors, nuclear transport inhibitors (selinexor), anti-apoptosis inhibitors, adoptive cell therapy, bispecific T cell engagers (BiTEs), B cell targeting agents and monoclonal antibodies.

[0462] Thus, in certain embodiments, the pharmaceutical formulations of the present invention comprise a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, and one or more additional therapeutic agents optionally selected from the group consisting of steroids (prednisone, prednisolone and dexamethasone), IMiDs (thalidomide, lenalidomide and pomalidomide), PIs (bortezomib, carfilzomib and ixazomib), histone deacetylase (HDAC) inhibitors (panobinostat), conventional chemotherapy (alkylating agents (e.g., melflufenazol, melphalan, cyclophosphamide, bendamustine), doxorubicin), checkpoint inhibitors, nuclear transport inhibitors (selinexor), anti-apoptosis inhibitors, adoptive cell therapy, bispecific T cell engagers (BiTEs), B cell targeting agents and monoclonal antibodies.

[0463] Preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, and one or more additional therapeutic agents selected from the group consisting of a proteasome inhibitor (PI), an immunomodulatory drug (IMiD), a B cell targeting agent, a steroid, a histone deacetylase (HDAC) inhibitor and a monoclonal antibody.

[0464] More preferably, the pharmaceutical formulation of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, and one or more additional therapeutic agents selected from the group consisting of a proteasome inhibitor (PI), a steroid and a monoclonal antibody.

[0465] For example, the pharmaceutical formulations of the present invention may consist essentially of a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, and one or more additional therapeutic agents. Alternatively, for example, the pharmaceutical formulations of the present invention may consist essentially of a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, one or more additional excipients, and one or more additional therapeutic agents.

[0466] The precise amount and concentration of the additional therapeutic agent that may be present in the pharmaceutical formulations of the present invention may vary with the dosage regimen, the potency of the specific agent selected, the age, size, sex and condition of the subject (typically a mammal, such as a human), the nature and severity of the disease or condition, and other relevant medical and physical factors. One skilled in the art can readily determine the amount and concentration of a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, and optionally one or more additional therapeutic agents suitable for use in the present invention.

[0467] treat

[0468] The pharmaceutical preparations, compositions and kits of the present invention can be used as medicines.

[0469] In certain embodiments, the pharmaceutical formulations, compositions, and kits of the invention are useful for treating and / or preventing diseases or conditions that are treatable with conventional chemotherapeutic agents, such as alkylating agents (eg, melphalan, cyclophosphamide, and bendamustine).

[0470] In certain embodiments, the pharmaceutical preparations and compositions of the present invention can be used to treat and / or prevent cancer, slow tumor growth, kill tumor cells, and / or treat and / or prevent amyloidosis. For example, the pharmaceutical preparations and compositions of the present invention can be used to treat and / or prevent cancer, slow tumor growth, and / or kill tumor cells. Thus, the pharmaceutical preparations and compositions of the present invention can be used to cure patients suffering from cancer and / or prolong their survival.

[0471] In an embodiment, the pharmaceutical preparations, compositions and kits of the present invention can be used to treat or prevent cancer.Specific examples of cancers that can be treated or prevented by administering the compounds or compositions of the present invention include carcinoma, sarcoma, myeloma, leukemia, lymphoma or mixed cancer.Exemplary cancers that can be treated or prevented by administering the compounds or compositions of the present invention include but are not limited to blood cancer / blood cell cancer, such as leukemia (e.g., acute lymphocytic leukemia, including adult and childhood acute lymphocytic leukemia; acute myeloid leukemia, including adult and childhood acute myeloid leukemia; chronic lymphocytic leukemia, such as B cell chronic lymphocytic leukemia; chronic myelogenous leukemia; and hairy cell leukemia); lymphoma (e.g., AIDS-related lymphoma; cutaneous T-cell lymphoma; Hodgkin's lymphoma, including adult and childhood Hodgkin's lymphoma and pregnancy Hodgkin's lymphoma; Non-Hodgkin's lymphoma, including adult and pediatric non-Hodgkin's lymphoma and non-Hodgkin's lymphoma during pregnancy; mycosis fungoides; Sézary syndrome; Waldenstrom's macroglobulinemia; primary mediastinal large B-cell lymphoma; mantle cell lymphoma; diffuse large B-cell lymphoma; and primary central nervous system lymphoma); and other hematologic cancers (e.g., chronic myeloproliferative disorders; multiple myeloma / plasma cell neoplasms; myelodysplastic syndromes; and myelodysplastic / myeloproliferative disorders); osteosarcoma; ovarian cancer; breast cancer; lung cancer; glioblastoma; retinoblastoma; and metastases of the foregoing cancers.

[0472] The pharmaceutical preparations, compositions and kits of the present invention are particularly cytotoxic to blood cancer cells. Therefore, the pharmaceutical preparations, compositions and kits of the present invention are particularly useful for treating or preventing one or more of the above-mentioned blood cancers.

[0473] In an embodiment, when the pharmaceutical formulations, compositions and kits of the present invention are used to treat or prevent cancer (e.g., selected from blood cancer / blood cell cancer, such as leukemia (e.g., acute lymphocytic leukemia, including adult and childhood acute lymphocytic leukemia; acute myeloid leukemia, including adult and childhood acute myeloid leukemia; chronic lymphocytic leukemia, such as B-cell chronic lymphocytic leukemia; chronic myelogenous leukemia; and hairy cell leukemia); lymphoma (e.g., AIDS-related lymphoma; cutaneous T-cell lymphoma; Hodgkin's lymphoma, including adult and childhood Hodgkin's lymphoma and pregnancy Hodgkin's lymphoma; non-Hodgkin's lymphoma, including adult and childhood non-Hodgkin's lymphoma and pregnancy non-Hodgkin's lymphoma; mycosis fungoides; Sezary syndrome; Waldenstrom's giant cell lymphoma); When the formulation or composition is used to treat a variety of hematologic malignancies (e.g., leukemia, primary mediastinal large B-cell lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma, and primary central nervous system lymphoma), and other hematologic malignancies (e.g., chronic myeloproliferative disorders, multiple myeloma / plasma cell neoplasms, myelodysplastic syndromes, and myelodysplastic / myeloproliferative disorders), osteosarcoma, ovarian cancer, breast cancer, lung cancer, glioblastoma, retinoblastoma, and metastases of the above cancers), the formulation or composition is administered (e.g., simultaneously, sequentially, or separately) with one or more additional therapeutic agents (e.g., steroids, IMiDs, PIs, HDAC inhibitors, conventional chemotherapy, checkpoint inhibitors, nuclear transport inhibitors, anti-apoptosis inhibitors, adoptive cell therapy, BiTEs, B cell targeting agents, and monoclonal antibodies).

[0474] The amount of a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate, required to achieve a therapeutic effect will vary with the specific route of administration and the characteristics of the subject being treated (e.g., species, age, weight, sex, medical condition, specific disease and its severity, and other relevant medical and physical factors). An ordinary skilled physician can readily determine and administer an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate, required for the treatment or prevention of cancer.

[0475] The pharmaceutical preparations and compositions of the present invention can be used in methods for treating subjects comprising administering a pharmaceutically effective amount of the pharmaceutical preparations or compositions of the present invention to the subject. Typically, the pharmaceutical preparations of the present invention are administered after being diluted into a physiologically acceptable solvent or diluent (e.g., saline solution or glucose solution).

[0476] Unit dose

[0477] The pharmaceutical preparations of the present invention may be provided in unit doses. Preferred unit doses are those containing the desired dose of a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, suitable for use according to the present invention.

[0478] In a preferred embodiment, the pharmaceutical formulations of the present invention may be provided in vials containing a unit dose of a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate. For example, the pharmaceutical formulations of the present invention can be provided in vials containing a unit dose of 1 to 800 mg of a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate (excluding the mass of any counterion). For example, the vial can contain a unit dose of 1 mg, 2 mg, 3 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg or 800 mg of a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate (excluding the mass of any counterion).

[0479] Reagent test kit

[0480] The present invention provides a kit suitable for preparing a pharmaceutical preparation according to the present invention. The kit of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, and a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin. The compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate and a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin in the kit can be provided together as a mixture, or they can be provided separately, for example, in separate vials, and then combined before using the kit (for example, before being combined to prepare a pharmaceutical preparation or composition of the present invention for use in the method or use of the present invention as defined herein).

[0481] In certain embodiments, the kit of the present invention further comprises one or more physiologically acceptable aqueous solvents or diluents, such as glucose solution or saline solution. Typically, the physiologically acceptable aqueous solvent or diluent is provided separately from the compound of formula (I) or its pharmaceutically acceptable salt, ester, amide or carbamate, or salt of such ester, amide or carbamate, and the negatively charged cyclodextrin or pharmaceutically acceptable derivative of the negatively charged cyclodextrin in the kit; or the physiologically acceptable aqueous solvent or diluent is provided together with the negatively charged cyclodextrin or pharmaceutically acceptable derivative of the negatively charged cyclodextrin as a mixture in the kit, and the compound of formula (I) or its pharmaceutically acceptable salt is provided separately. , ester, amide or carbamate, or a salt of such an ester, amide or carbamate, is provided separately, for example, a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate is provided in a separate vial and then combined with a composition comprising a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin and a physiologically acceptable aqueous solvent or diluent prior to use of the kit (for example, combined prior to use in the methods or uses of the invention as defined herein to prepare a composition of the invention).

[0482] In one embodiment of the invention, the kit of the invention further comprises one or more other therapeutic agents as described herein. The one or more other therapeutic agents in the kit may be provided as a mixture with one or both of a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate and a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, or may be provided separately.

[0483] Preferably, the kit of the present invention comprises a compound of formula (I) as described herein, or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, in the form of a lyophilized formulation comprising a compound of formula (I) as described herein, or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, and sucrose (or consisting essentially of the same).

[0484] For the avoidance of doubt, the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, the negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, optionally a physiologically acceptable aqueous solvent or diluent, optionally one or more pharmaceutically acceptable organic solvents and optionally one or more other therapeutic agents are present in the kit according to the invention in a form and amount suitable for the preparation of a pharmaceutical formulation according to the invention. The amount of the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, the pharmaceutically acceptable organic solvent and optionally one or more other therapeutic agents suitable for use according to the invention can be readily determined by those skilled in the art.

[0485] The present invention also provides a kit suitable for preparing injections and infusions, comprising a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such ester, amide or carbamate, and a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin. Such kits comprise a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, and one or more physiologically acceptable aqueous solvents or diluents, such as glucose solution or saline solution. A compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, and one or more physiologically acceptable aqueous solvents or diluents can be provided as a mixture or provided separately. For example, a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, and a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin may be provided together as a mixture, and one or more physiologically acceptable aqueous solvents or diluents may be provided separately. Alternatively, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin and one or more physiologically acceptable aqueous solvents or diluents may be provided as a mixture (i.e., as a composition comprising a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin and one or more physiologically acceptable aqueous solvents or diluents), and a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, may be provided separately.

[0486] Preferably, the pharmaceutically acceptable aqueous solvent is a 5% glucose solution (w / v in water) or a 0.9% sodium chloride solution (w / v in water). Preferably, such solutions are suitable for intravenous injection and / or infusion into a subject.

[0487] In such a kit, preferably, the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, and a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin are provided in the form of a pharmaceutical preparation of the present invention (e.g., a lyophilized powder of the present invention or a liquid pharmaceutical formulation of the present invention). Thus, the present invention provides a kit suitable for preparing an injection and an infusion, comprising a pharmaceutical preparation of the present invention (e.g., a lyophilized powder of the present invention) and one or more physiologically acceptable aqueous solvents or diluents as described herein, such as a glucose solution or a saline solution.

[0488] Alternatively, the present invention provides a kit suitable for preparing injections and infusions, comprising: a pharmaceutical preparation comprising a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate (for example, a lyophilized pharmaceutical preparation comprising a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate and optionally sucrose); a composition comprising a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin; and one or more physiologically acceptable aqueous solvents or diluents as described herein, such as a glucose solution or a saline solution.

[0489] Alternatively, the present invention provides a kit suitable for preparing injections and infusions, comprising: a pharmaceutical preparation comprising a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate (for example, a lyophilized pharmaceutical preparation comprising a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate and optionally sucrose), and a composition comprising a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin and one or more physiologically acceptable aqueous solvents or diluents as described herein (for example, a glucose solution or a saline solution).

[0490] In one embodiment of the invention, the kit of the invention suitable for the preparation of injections and infusions further comprises one or more other therapeutic agents as described herein.

[0491] Preferably, the kit of the present invention comprises instructions, e.g., instructions directing the user to mix a specified amount of a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, with a specified amount of a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin.

[0492] Such instructions may also provide guidance regarding storage conditions and / or administration instructions.

[0493] For the avoidance of doubt, the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, one or more physiologically acceptable aqueous solvents or diluents and optionally one or more other therapeutic agents are present in the kit according to the present invention in a form and amount suitable for the preparation of a solution suitable for use as an injection or infusion. The amount of the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, a physiologically acceptable aqueous solvent and optionally one or more other therapeutic agents suitable for use according to the present invention can be readily determined by those skilled in the art.

[0494] Finally, in embodiments of the invention providing a kit, the kit of the invention may comprise a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the weight ratio (w / w) of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate is from about 1:1 to about 100:1, for example, comprising the following negatively charged cyclodextrin or a pharmaceutically acceptable derivative of the negatively charged cyclodextrin: The invention relates to a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the weight ratio (w / w) of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate is from about 4:1 to about 80:1, from about 2:1 to about 50:1 or from about 5:1 to about 20:1, wherein the amount of the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate is the amount present in the kit. For example, the kit of the present invention may comprise a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, wherein the weight ratio (w / w) of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate is about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1 or about 25:1. In a preferred embodiment, the weight ratio (w / w) of the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin to the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate is about 5:1, about 10:1 or about 20:1, wherein the amount of the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate, or a salt of such an ester, amide or carbamate is the amount present in the kit.

[0495] Equivalent

[0496] The present invention is described broadly and generally herein. A person of ordinary skill in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are exemplary only, and that the actual parameters, dimensions, materials, and / or configurations will depend on the specific application or applications for which the teachings of the present invention are used. A person of ordinary skill in the art will recognize or be able to determine, using only routine experimentation, many equivalents to the specific embodiments of the present invention described herein. Therefore, it should be understood that the foregoing embodiments are presented by way of example only, and that, within the scope of the appended claims and their equivalents, the present invention may be practiced in a manner different from that specifically described and claimed. The present invention relates to each individual feature, system, article, material, kit, and / or method described herein. In addition, any combination of two or more such features, systems, articles, materials, kits, and / or methods is included within the scope of the present invention if it is not mutually inconsistent. In addition, each of the narrower types and subgeneric groups falling within the general disclosure also forms part of the present invention. This includes the general description of the present invention, with the proviso or negative limitation that any subject matter is removed from it, regardless of whether the deleted material is specifically described herein.

[0497] Incorporated by Reference

[0498] The contents of the articles, patents, and patent applications mentioned or cited herein, and all other documents and electronically available information are hereby incorporated by reference in their entirety to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference. Applicants reserve the right to physically incorporate into this application any and all materials and information from any of these articles, patents, patent applications, or other physical and electronic documents.

[0499] The following examples illustrate the invention.

[0500] Example

[0501] The following exemplary compounds and biological examples are disclosed in International Patent Application No. PCT / EP2022 / 066756, published as WO 2022 / 263679. The exemplary formulations illustrate the pharmaceutical formulations claimed herein.

[0502] Example compounds

[0503] The compounds described herein can be prepared according to methods known to those skilled in the art. Other reaction schemes and various solvents, temperatures and other reaction conditions can be easily designed by those skilled in the art.

[0504] The compounds of formula (I) can be prepared according to the scheme described in International Patent Application No. PCT / EP2022 / 066756, which is published as WO2022 / 263679. The synthetic methods of exemplary compounds 1 to 51 described on pages 37 to 132 of WO2022 / 263679 are incorporated herein by reference. For completeness, the structures of the compounds are shown in Table 1 below.

[0505] Biological Examples

[0506] Biological Example 1—Evaluation of In Vitro Cytotoxic Potency in MM.1S Cell Viability Assay:

[0507] Methods: MM.1S cells were cultured in RPMI medium 1640 (IX) supplemented with 10 μg / μL penicillin / streptomycin, 10% heat-inactivated FBS, 0.1 mM sodium pyruvate, 1x MEM NEAA (ThermoFisher), and 1 mM HEPES. Cells were grown to logarithmic phase before exposure to test compounds.

[0508] Test compounds (exemplary compounds 1 to 36, 38 to 41, and 45 to 48) and comparative compounds (bendamustine, busulfan, melphalan, chlorambucil, and 4-hydroperoxycyclophosphamide) were dissolved in 100% DMSO and added to a 384-well assay plate. Additional DMSO was added to maintain the same final DMSO concentration (0.2%) in all wells. 25 μL of a cell suspension at a concentration of 100,000 cells / ml was added to each well. The cells were cultured in a CO2 incubator at 37°C for 72 hours. The plate was equilibrated to room temperature for 30 minutes, and then 20 μL of 2.0 (Promega) reagent. The plates were then allowed to stabilize for 20 minutes before recording luminescence in an Envision plate reader (emission filter 700 nm). Plate reader data were normalized to negative controls (untreated) and positive controls (cells treated with 400 μM chlorpromazine for 72 hours), and IC values ​​were derived from a 4-parameter logistic regression curve fit. 50 .

[0509] Results: The test compounds were evaluated for their ability to inhibit MM.1S cell proliferation and induce in vitro cytotoxicity. Dose responses can be extracted from the experiments, as shown in Table 1 below. The test compounds exhibited good cytotoxicity against MM.1S cells. In this assay, exemplified compounds 1 to 16, 21 to 32, 34 to 36, 38, and 45 to 48 were found to be more potent cytotoxic than bendamustine, busulfan, melphalan, chlorambucil, and 4-hydroperoxycyclophosphamide.

[0510] Table 1. Cytotoxicity to MM.1S cells

[0511]

[0512]

[0513]

[0514]

[0515]

[0516]

[0517]

[0518]

[0519]

[0520]

[0521]

[0522]

[0523] Biological Example 2—Evaluation of in vitro cytotoxicity on blood cell lines, human peripheral blood mononuclear cells and normal fibroblasts:

[0524] Methods: The cell lines used in these experiments are listed in Table 2 below. All cells were cultured in RPMI medium 1640 (IX). All media were supplemented with 10 μg / μl penicillin / streptomycin, 10% heat-inactivated FBS, 0.1 mM sodium pyruvate, 1x MEMNEAA (ThermoFisher), and 1 mM HEPES. DERL-2 cells had a higher FBS (20%) and were supplemented with 40 ng / mL IL-2. hPBMCs were thawed and acclimated overnight, then stimulated with 10 μg / mL PHA-M and 40 ng / mL IL-2 for 4 hours before exposure to test compounds. All cells, except hPBMCs, were grown to logarithmic phase before exposure to test compounds.

[0525] Table 2. Cell lines used in in vitro cytotoxicity experiments

[0526]

[0527]

[0528] The test compound was dissolved in 100% DMSO and added to a 384 assay plate. Additional DMSO was added to maintain the same final DMSO concentration (0.2%) in all wells. 25 μL of a cell suspension at a concentration of 100,000 cells / mL was added to each well. The cells were cultured in a 37°C CO2 incubator for 72 hours. The plate was equilibrated to room temperature for 30 minutes, and then 20 μL of 2.0 (Promega) reagent. The plates were then allowed to stabilize for 20 minutes before recording luminescence in an Envision plate reader (emission filter 700 nm). Plate reader data were normalized to negative controls (untreated) and positive controls (cells treated with 400 μM chlorpromazine for 72 hours), and IC values ​​were derived from a 4-parameter logistic regression curve fit. 50 .

[0529] Results: The ability of the test compounds to inhibit cell proliferation in vitro was evaluated using several different hematological cell lines. Cytotoxicity was also assessed in non-malignant human peripheral blood mononuclear cells (hPBMC) and fibroblasts (BJ). Table 3 shows the dose response of all test compounds extracted from the experiments. In the table, the results are expressed as cytotoxicity IC50 μM + / - standard deviation (number of tests). nd = value not determined.

[0530] Table 3a and 3b. Cytotoxicity of test compounds against normal and cancer cell lines

[0531] Table 3a:

[0532]

[0533] Table 3b

[0534]

[0535] It can be seen that the cytotoxicity of the compound of formula (I) is consistently greater than that of the two compounds of the prior art.

[0536] Biological Example 3 - Evaluation of Cellular Retention / Efflux of Example 18 (after treatment with Example 1), Bendamustine, and Melphalan:

[0537] Method: 2×10 6 MM.1S cells were seeded at 250 μL per well in a 96-well plate at 10 cells / mL. Cells were incubated in the presence of test compounds for 5, 15, and 60 minutes, then centrifuged at +4°C for 5 minutes to pellet the cells. Cell culture medium was immediately collected onto a second plate and stored at -80°C until further analysis. The pelleted cells were washed once with ice-cold PBS and then stored at -80°C until further analysis.

[0538] The cells were lysed and proteins in the culture medium were precipitated using acetonitrile: dimethylformamide (9:1) containing an internal standard. The samples were centrifuged and the supernatant was analyzed by LC-MS / MS (ACQUITY UPLC-Xevo TQ-Smicro) to determine compound concentrations.

[0539] As test compounds, Exemplary Compound 1, bendamustine, and melphalan were used.

[0540] Results: The results are shown in Figure 1. Figure 1 (A) shows the intracellular and extracellular concentrations of bendamustine and a metabolite compound (Example Compound 18) formed from Example Compound 1. It can be seen that after treatment with Example Compound 1, Example Compound 18 is rapidly formed inside the cells, while its content in the culture medium outside the cells is low. These results indicate that the hydrolysis of Example Compound 1 occurs inside the cells and that the metabolites of Example Compound 1 remain inside the cells. Low intracellular concentrations of bendamustine can be detected, and its level decreases over time. The extracellular concentration of bendamustine is quite high and decreases over time.

[0541] Figure 1(B) shows the intracellular and extracellular concentrations of melphalan during the time course of the experiment. After in vitro treatment of MM.1S cells with melphalan, low intracellular concentrations of melphalan were detected. The extracellular concentration of melphalan was high and constant throughout the experiment.

[0542] Biological Example 4—In vivo evaluation of the cytotoxicity of test compounds in a chicken embryo xenograft model:

[0543] Method: On the 9th day (after fertilization), 3×10 6 Individual SU-DHL-4 tumor cells were inoculated into fertilized white Leghorn eggs. On days 11, 13, 15, and 17, surviving eggs were injected with 100 μL of Example Compound 1 (8.4 μM (0.008 mg / kg), 33.5 μM (0.033 mg / kg), 167.5 μM (0.164 mg / kg)), bendamustine (8.4 μM (0.005 mg / kg), 33.5 μM (0.02 mg / kg), 167.5 μM (0.1 mg / kg)), or vehicle. On day 18, tumors were excised and weighed.

[0544] Results: Example compound 1 had a significant dose-dependent effect on tumor weight ( Figure 2For example compound 1, tumor reduction rates compared to vehicle were 38%, 70%, and 82% at 8.4 μM, 33.5 μM, and 167.5 μM, respectively. For bendamustine, tumor reduction rates were only significant at 33.5 μM and 167.5 μM, representing 30% and 76% reductions, respectively, compared to vehicle. Therefore, example compound 1 exhibited potent tumor cytotoxicity in this model.

[0545] Biological Example 5—Evaluation of Cellular Retention / Efflux of PDC Compounds and Metabolites:

[0546] Methods: The methods were as described in Biological Example 3.

[0547] Example compounds 1, 2, 3, 4, 6, 7 and 10 were studied. All tested compounds form intracellular metabolites. The compounds used for cell treatment are metabolized into ester hydrolysis compounds (referred to herein as metabolite A) and amide hydrolysis compounds (referred to as metabolite B). The structures of the example compounds and the structures of the two metabolites of the corresponding compounds are shown in Table 4. The metabolite B formed under the test conditions is a compound having the structure of Examples 17 and 18:

[0548]

[0549] Table 4:

[0550]

[0551]

[0552]

[0553] The intracellular concentration of the metabolites slowly decreased over time. Little or no extracellular concentration of the metabolites could be detected in the culture medium outside the cells. These results indicate that hydrolysis of the example compounds occurs inside the cells and that the metabolites of the example compounds remain inside the cells.

[0554] Biological Example 6—Evaluation of Direct DNA Damage by Test Compounds:

[0555] Methods: DNA was prepared from MM.1S cells using a QIAGEN genomic tip 20 / G according to the protocol described in Furda et al., Methods Mol Biol, 2012. 1 μg of DNA was treated with 0.25 μM of compound at 37°C for 30 minutes. DMSO was used as a control treatment. 20 ng of treated DNA was PCR amplified using Phusion Hot Start II High-Fidelity PCR Master Mix (Thermo Scientific) using primers targeting HPRT or primers targeting mitochondrial DNA (mtDNA) to generate fragments of 10.4 or 8.9 kb, respectively. PCR reactions were separated on 1% agarose gels and analyzed by ChemiDoc. TM The MP imaging system (BioRad) was used for visualization. Example compounds 1, 2, and 38, as well as metabolites of compound 1, compounds 18 and 20, were studied. They were compared with bendamustine, melphalan, and DMSO controls. The results for compounds 1, 2, 18, and 20 using HPRT primers are shown in Figure 2. Figure 4 (A) shows the results; the results using mtDNA primers are shown in Figure 4 (B) The results of compound 18 and 38 using HPRT primer and mtDNA primer are shown as follows Figure 4 (C) shown.

[0556] Results: From Figure 4 As can be seen in the table, at the tested concentrations, melphalan and bendamustine had little or no effect on DNA damage to nuclear and mitochondrial DNA. Example compounds 1 and 2 had strong DNA-damaging effects on both nuclear and mitochondrial DNA, as indicated by the absence of PCR bands detected on the gel. Example compounds 18 and 38 effectively damaged both nuclear and mitochondrial DNA, with only faint PCR bands detected. Example compound 20 also had DNA-damaging effects, but to a lesser extent than Example compounds 1, 2, and 18, but greater than that of bendamustine or melphalan.

[0557] Biological Example 7—Evaluation of DNA Fragmentation Induced by Test Compounds

[0558] Method: 1×10 6MM.1S cells (CRL-2974, ATCC) were treated with 0.006, 0.06, 0.6 or 6 μM of example compound 1 or bendamustine for 24 hours. After incubation, the cells were washed with PBS (10010-015, Gibco), fixed with flow cytometry fixative (FC004, R&D Systems) for 10 minutes, and stored in 70% ethanol at -20°C overnight. Deoxythymidine analog (BrdUTP) was present in the experiment to mark DNA break sites. The next day, APO-BrdU was used according to the manufacturer's protocol (A23210, Invitrogen). TM Cells were washed and stained with TUNEL assay kit and then stained with BD FACSCanto TM II was analyzed. The percentage of TUNEL-positive cells indicates the percentage of cells with fragmented DNA.

[0559] Results: The dose-response effects of Example Compound 1 and bendamustine on DNA damage after 24 hours of incubation are shown in Figure 5(A). All data are presented as mean ± SD (n = 2). 24-hour exposure to Example Compound 1 caused DNA damage in a dose-dependent manner, while bendamustine had no significant effect on DNA damage after 24 hours of incubation. Representative FACS histograms (n = 2) of Alexa fluor 488 intensity after 24 hours of treatment with 0.6 μM test compound or bendamustine are shown in Figure 5(B). The top pair of FACS histograms is Example Compound 1; the middle pair is bendamustine; and the bottom pair is the DMSO control. Notably, 24-hour treatment with 0.6 μM Example Compound 1 resulted in DNA breaks in >90% of the tested cells, whereas bendamustine did not. Example Compound 1 was more potent in causing DNA damage.

[0560] Example of formulation:

[0561] Example Preparation 1

[0562] Example compound 1*: 20 mg (free base)

[0563] Sodium sulfobutyl ether beta-cyclodextrin 200mg

[0564] Water for injection 10mL

[0565] *(2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester

[0566] Example Formulation 2 (pH 4.5)

[0567]

[0568] Example Formulation 3 (pH 4.9)

[0569]

[0570]

[0571] Stability of Example Compound 1 with Different Excipients

[0572] (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 1; 19.2 mg, 16.7 mg free base) was dissolved in dimethylacetamide (419 μL) to give a 40 mg / mL stock solution.

[0573] As shown in Table 5, a 0.4 mg / mL solution of Example Compound 1 was prepared by diluting a dimethylacetamide stock solution (40 mg / mL, 10 μL) with various solutions (990 μL), and the stability after 4 hours at room temperature was analyzed by HPLC (analytical HPLC was performed on an Agilent Series 1100 system using a Kinetex XB C18 (2.6 μm, 3.0×50 mm) column and a mobile phase of 0.1% TFA in H 2 O / CH 3 CN (acidic conditions) (flow rate 1 mL / min). Excipients are reported as the weight ratio of excipient to Example Compound 1 (free base).

[0574] Table 5:

[0575]

[0576]

[0577] 1 Dulbecco's phosphate buffered saline: poor solubility.

[0578] Experiments showed that in ultrapure water, the best excipient was sodium sulfobutyl ether beta-cyclodextrin, in which very little material degraded after four hours at room temperature.

Claims

1. A pharmaceutical formulation comprising a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin and a compound according to formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate in, X is C 1-6 alkylene; W1, W2, W3 and W4 are each CH, or one of W1, W2, W3 and W4 is N and the rest are CH; R 1 is selected from the group consisting of: H; C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-4 alkyl; and halogen; R 2 is selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-6 alkyl; and R 3 is a group according to formula (II): or a group according to formula (III): in, R 4 Selected from the group consisting of: N(R c )(R d ) and formula (IV): and When R 4 When it is formula (IV), R 5 R b ; and when R 4 N(R c )(R d ), R 5 Selected from the group consisting of: R b Japanese style (V) Among them, each R a Independently selected from the group consisting of: H; C 1-6 Alkyl; -CH2-phenyl; or -CH2-3 to 12-membered heterocyclic group containing 1, 2, 3 or 4 heteroatoms selected from N, O and S; wherein The C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of: -OH, -OC 1-6 alkyl, -NH2, -NHC(=NH)NH2, -C(O)OH, -C(O)NH2, -SH, -SCH3 and halogen; and the phenyl or heterocyclic group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halogen, -NH2, -OH, -OC 1-6 Alkyl and -NO2; R b Selected from the group consisting of: -OH; -N(R e )(R f ); and -OC optionally substituted by one or more groups selected from the following 1-6 Alkyl: halogen, -OH, -CN, -N(R e )(R f ),-C 6-10 aryl or a 3- to 12-membered heterocyclic ring comprising one or more O, N or S atoms and optionally substituted by 1, 2 or 3 halogens, and / or wherein the alkyl is optionally interrupted by 1, 2 or 3 O, N or S atoms; R c and R d Each independently selected from the group consisting of: H; -C 1-6 Alkyl; C(O)C 1-6 alkyl; and -CH2-phenyl, wherein the alkyl or the phenyl is optionally substituted with 1, 2 or 3 groups selected from halogen; and R e and R f Each independently selected from the group consisting of: H and -C 1-6 Alkyl, wherein the alkyl is optionally substituted with 1, 2 or 3 groups selected from halogen; or R e and R f Together with the nitrogen atom to which they are attached they form a 4-, 5- or 6-membered heterocyclic ring which is optionally substituted by 1, 2 or 3 groups selected from halogen.

2. The pharmaceutical formulation according to claim 1, wherein the compound of formula (I) or the pharmaceutically acceptable salt, ester, amide or carbamate thereof, or the salt of such ester, amide or carbamate has structure (Ia): For example, structure (Ib):

3. The pharmaceutical preparation according to claim 1 or claim 2, wherein R 1 For H.

4. The pharmaceutical preparation according to any one of claims 1 to 3, wherein R 2 is selected from the group consisting of: phenyl optionally substituted by 1, 2 or 3 halogens; and -C 1-6 Alkyl; for example -C optionally substituted by 1, 2 or 3 halogen 1-4 Alkyl groups, such as methyl.

5. The pharmaceutical formulation according to any one of claims 1 to 4, wherein X is -CH2- or -CH2-CH2-.

6. The pharmaceutical preparation according to any one of claims 1 to 5, wherein R 3 is a group according to formula (IIa): or a group according to formula (IIIa):

7. The pharmaceutical preparation according to any one of claims 1 to 6, wherein R 4 Selected from the group consisting of: N(R c )(R d ) and formula (IVa):

8. The pharmaceutical preparation according to any one of claims 1 to 7, wherein when R 4 When it is formula (IV) or (IVa), R 5 R b ; and when R 4 N(R c )(R d ), R 5 Selected from the group consisting of: R b and formula (Va), wherein formula (Va) has the following structure, 9. The pharmaceutical preparation according to any one of claims 1 to 8, wherein R 4 is NH2.

10. The pharmaceutical preparation according to any one of claims 1 to 9, wherein each R a Independently selected from the group consisting of: C 1-6 Alkyl and -CH2-phenyl; wherein the C 1-6 The alkyl group is optionally substituted with: -OH; -NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen (e.g., F or Cl); and the phenyl group is optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of: halogen (e.g., F or Cl); -NH2; -OH; -OC 1-6 Alkyl; and -NO2.

11. The pharmaceutical preparation according to claim 10, wherein each R a Selected from the group consisting of: -C 1-6 alkyl and -CH2-phenyl; wherein the phenyl is optionally substituted with 1, 2 or 3 halogen substituents.

12. The pharmaceutical preparation according to any one of claims 1 to 11, wherein when R 4 When it is formula (IV) or (IVa), R 5 For-OC 1-6 alkyl; and when R 4 When it is NH2, R 5 For-OC 1-6 Alkyl or formula (V) or (Va).

13. The pharmaceutical preparation according to claim 12, wherein when R b For-OC 1-6 When alkyl, it is selected from the group consisting of methoxy, ethoxy and isopropoxy.

14. The pharmaceutical formulation according to claim 1, wherein the compound of formula (I) or the pharmaceutically acceptable salt, ester, amide or carbamate thereof, or the salt of such ester, amide or carbamate is selected from the group consisting of: (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 1); (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-(4-fluorophenyl)propanoic acid ethyl ester (Example Compound 2); (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-methyl-butyric acid ethyl ester (Example Compound 3); (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 4); (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-3-(4-fluorophenyl)propionic acid ethyl ester (Example Compound 5); (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-3-methyl-butyric acid ethyl ester (Example Compound 6); (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 7); (2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propionyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Exemplary Compound 8); (2S)-2-[[(2S)-2-amino-3-methyl-butyryl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 9); (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionic acid ethyl ester (Example Compound 10); (2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propionyl]amino]-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionic acid ethyl ester (Example Compound 11); (2S)-2-[[(2S)-2-amino-3-methyl-butyryl]amino]-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionic acid ethyl ester (Example Compound 12); (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-phenyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 13); (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid methyl ester (Example Compound 14); (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid isopropyl ester (Exemplary Compound 15); (2S)-2-[[(2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propionyl]amino]-4-methyl-pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 16); (2S)-2-[[(2R)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propionyl]amino]-4-methyl-pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 21) (2S)-2-[[(2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoyl]amino]-3-(4-fluorophenyl)propionic acid ethyl ester (Example Compound 22) (2S)-2-[[(2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 23) (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid 2-morpholinoethyl ester (Example Compound 24) (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid 2-isopropoxyethyl ester (Example Compound 25) (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-valeric acid isopropyl ester (Example Compound 26) (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid methyl ester (Example Compound 27) (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid 3-(dimethylamino)propyl ester (Example Compound 28) (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (2-methoxy-1-methyl-ethyl ester) (Example Compound 29) (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-N,N,4-trimethyl-pentanamide (Example Compound 30) (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-N,4-dimethyl-pentanamide (Example Compound 31) (2S)-2-[[(2S)-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]-2-(methylamino)butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 32) (2S)-2-[[(2S)-2-acetylamino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-(4-fluorophenyl)propionic acid ethyl ester (Example Compound 34) (2S)-2-[[(2S)-2-Acetylamino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 35) (2S)-2-[[(2S)-2-acetylamino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-3-(4-fluorophenyl)propionic acid ethyl ester (Example Compound 36) (2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 38) (2R)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 45) (2S)-2-[[(2R)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 46) (2R)-2-[[(2R)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 47) (2S)-2-[[(2S)-2-amino-4-[6-[bis(2-chloroethyl)amino]-3-methyl-imidazo[4,5-b]pyridin-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Exemplary Compound 48); and (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloro-1,1,2,2-tetradeuterium-ethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid ethyl ester (Example Compound 50) or a pharmaceutically acceptable salt, amide or carbamate thereof, including salts of such amides or carbamates.

15. The pharmaceutical formulation according to claim 1, wherein the compound of formula (I) or the pharmaceutically acceptable salt, ester, amide or carbamate thereof, or the salt of such ester, amide or carbamate is selected from the group consisting of: (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-methyl-butanoic acid (Example Compound 19); and (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (Example Compound 20); (2S)-2-[[(2S)-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]-2-(methylamino)butanoyl]amino]-4-methyl-pentanoic acid (Example Compound 33) (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid (Example Compound 37) (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-3-(4-fluorophenyl)propanoic acid (Example Compound 39) (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-phenyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (Example Compound 40) 2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-3-methyl-butanoic acid (Example Compound 42) (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propanoic acid (Example Compound 43) (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propionyl]amino]-4-methyl-pentanoic acid (Exemplary Compound 44); and (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloro-1,1,2,2-tetradeuterium-ethyl)amino]-1-methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (Example Compound 51) or a salt thereof.

16. A pharmaceutical formulation comprising a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin and a compound according to formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate in, X is C 1-6 alkylene; W1, W2, W3 and W4 are each CH, or one of W1, W2, W3 and W4 is N and the rest are CH; R 1 is selected from the group consisting of: H; C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-4 alkyl; and halogen; R 2 is selected from the group consisting of: H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and C optionally substituted by 1, 2 or 3 groups independently selected from halogen 1-6 alkyl; and R 3 Formula (VIa): where R c and R d Each independently selected from the group consisting of: H, -C 1-6 Alkyl, C(O)C 1-6 alkyl and -CH2-phenyl, wherein the alkyl or the phenyl is optionally substituted with 1, 2 or 3 groups selected from halogen; and R g Selected from the group consisting of: H and -C 1-6 Alkyl, wherein the alkyl is optionally substituted with 1, 2 or 3 groups selected from halogen.

17. The pharmaceutical preparation according to claim 16, wherein the compound of formula (I) or the pharmaceutically acceptable salt, ester, amide or carbamate thereof, or the salt of such ester, amide or carbamate has the structure (Ia) or (Ib) shown in claim 2, or R 1 、R 2 or X as claimed in any one of claims 3 to 5; R c and R d Each is H; and R g For H.

18. The pharmaceutical formulation according to claim 16, wherein the compound of formula (I) or the pharmaceutically acceptable salt, ester, amide or carbamate thereof, or the salt of such ester, amide or carbamate is selected from the group consisting of: (2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]propanoic acid (Example Compound 17); (S)-2-amino-4-(5-(bis(2-chloroethyl)amino)-1-methyl-1H-benzo[d]imidazol-2-yl)butanoic acid (Example Compound 18); (2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid ethyl ester (Example Compound 38) (2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-1-phenyl-benzimidazol-2-yl]butanoic acid (Exemplary Compound 41); and (2S)-2-amino-4-[5-[bis(2-chloro-1,1,2,2-tetradeuterium-ethyl)amino]-1-methyl-benzimidazol-2-yl]butanoic acid (Example Compound 49); or a salt thereof.

19. A pharmaceutical formulation according to any one of the preceding claims, wherein the compound of formula (I) or an ester, amide or carbamate thereof is present in the form of a pharmaceutically acceptable salt.

20. The pharmaceutical formulation according to claim 19, wherein the compound of formula (I) or its ester, amide or carbamate is in the form of a hydrochloride salt.

21. A pharmaceutical formulation according to any one of the preceding claims comprising a pharmaceutically acceptable derivative of a negatively charged cyclodextrin.

22. The pharmaceutical formulation of claim 21, wherein the pharmaceutically acceptable derivative of the negatively charged cyclodextrin is a sulfoalkyl ether derivatized cyclodextrin.

23. The pharmaceutical formulation of claim 22, wherein the pharmaceutically acceptable derivative of the negatively charged cyclodextrin is a sulfobutyl ether derivatized cyclodextrin.

24. A pharmaceutical formulation according to claim 22 or claim 23, wherein the average number of sulfoalkyl groups per cyclodextrin ring, expressed as the average degree of substitution, is from 5 to 8.

25. The pharmaceutical formulation according to any one of the preceding claims, wherein the negatively charged cyclodextrin or pharmaceutically acceptable derivative of a negatively charged cyclodextrin is negatively charged β-cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged β-cyclodextrin.

26. The pharmaceutical formulation according to any one of the preceding claims, wherein the negatively charged cyclodextrin or pharmaceutically acceptable derivative of a negatively charged cyclodextrin is sodium sulfobutyl ether beta-cyclodextrin.

27. A pharmaceutical formulation according to any one of the preceding claims, comprising citric acid and / or one or more citrate salts (eg trisodium citrate dihydrate).

28. The pharmaceutical formulation of any one of the preceding claims, wherein the weight ratio (w / w) between the negatively charged cyclodextrin or the pharmaceutically acceptable derivative of the negatively charged cyclodextrin and the compound of formula (I) or the pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate (excluding the mass of any counterion) is 1:1 to 100:1, 2:1 to 50:1 or 5:1 to 20:

1.

29. The pharmaceutical formulation according to any one of the preceding claims, wherein the pharmaceutical formulation is a lyophilized powder.

30. The pharmaceutical formulation of claim 29, wherein the pharmaceutical formulation comprises 1 mg to 150 mg of the compound of formula (I) or the pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate (excluding the mass of any counterion).

31. A pharmaceutical formulation according to claim 29 or claim 30 which is substantially free of organic solvents.

32. The pharmaceutical formulation of any one of the preceding claims, comprising one or more additional therapeutic agents selected from the group consisting of steroids, immunomodulatory drugs (IMiDs), proteasome inhibitors (PIs), histone deacetylase (HDAC) inhibitors, conventional chemotherapy, checkpoint inhibitors, nuclear transport inhibitors, anti-apoptosis inhibitors, adoptive cell therapy, bispecific T cell engagers (BiTEs), B cell targeting agents, and monoclonal antibodies.

33. A composition comprising a pharmaceutical formulation according to any one of the preceding claims and a physiologically acceptable aqueous solvent or diluent.

34. A composition comprising a compound of formula (I) as defined in any one of claims 1 to 20 or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, and a physiologically acceptable aqueous solvent.

35. The composition of claim 33 or claim 34, wherein the physiologically acceptable aqueous solvent is a glucose solution, a saline solution, or a mixture thereof.

36. A kit comprising a pharmaceutical formulation according to any one of claims 1 to 32 and a physiologically acceptable aqueous solvent or diluent.

37. A kit comprising a compound of formula (I) as defined in any one of claims 1 to 20 or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, a negatively charged cyclodextrin or a pharmaceutically acceptable derivative of a negatively charged cyclodextrin, and optionally one or more physiologically acceptable aqueous solvents or diluents.

38. The kit of claim 37, wherein the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, is provided in the form of a lyophilized formulation comprising (or essentially consisting of): the compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, or a salt of such an ester, amide or carbamate, and optionally sucrose.

39. The pharmaceutical formulation according to claims 1 to 32, or the composition according to claims 33 to 35 or the kit according to claims 36 to 38 for use as a medicament.

40. The pharmaceutical formulation according to claims 1 to 32, or the composition according to claims 33 to 35, or the kit according to claims 36 to 38, for use in treating or preventing cancer.

41. The pharmaceutical formulation, composition, or kit for use according to claim 40, wherein the cancer is selected from the group consisting of: a blood cancer / blood cell cancer, such as a leukemia (e.g., acute lymphocytic leukemia, including adult and childhood acute lymphocytic leukemia; acute myeloid leukemia, including adult and childhood acute myeloid leukemia; chronic lymphocytic leukemia, such as B-cell chronic lymphocytic leukemia; chronic myelogenous leukemia; and hairy cell leukemia); a lymphoma (e.g., AIDS-related lymphoma; cutaneous T-cell lymphoma; Hodgkin's lymphoma, including adult and childhood Hodgkin's lymphoma and gestational Hodgkin's lymphoma); The present invention also includes the following types of cancer: non-Hodgkin's lymphoma, including adult and pediatric non-Hodgkin's lymphoma and non-Hodgkin's lymphoma during pregnancy; mycosis fungoides; Sézary syndrome; Waldenstrom's macroglobulinemia; primary mediastinal large B-cell lymphoma; mantle cell lymphoma; diffuse large B-cell lymphoma; and primary central nervous system lymphoma); and other hematologic cancers (e.g., chronic myeloproliferative disorders; multiple myeloma / plasma cell neoplasms; myelodysplastic syndromes; and myelodysplastic / myeloproliferative disorders); osteosarcoma; ovarian cancer; breast cancer; lung cancer; glioblastoma; retinoblastoma; and metastases of the above cancers.

42. A pharmaceutical formulation, composition, or kit for use according to claim 40 or 41, wherein the formulation or composition is administered (e.g., simultaneously, sequentially, or separately) with one or more additional therapeutic agents, such as steroids, IMiDs, PIs, HDAC inhibitors, conventional chemotherapy, checkpoint inhibitors, nuclear transport inhibitors, anti-apoptosis inhibitors, adoptive cell therapy, BiTEs, B cell targeting agents, and monoclonal antibodies.

43. A method for treating a patient, said method comprising administering a pharmaceutically effective amount of a pharmaceutical formulation according to claims 1 to 32 or a composition according to claims 33 to 35.

44. A method for treating or preventing cancer, comprising administering an effective amount of a pharmaceutical formulation according to claims 1 to 32 or a composition according to claims 33 to 35, e.g., wherein the cancer is selected from the group consisting of: a blood cancer / blood cell cancer, e.g., a leukemia (e.g., acute lymphocytic leukemia, including adult and childhood acute lymphocytic leukemia; acute myeloid leukemia, including adult and childhood acute myeloid leukemia; chronic lymphocytic leukemia, such as B-cell chronic lymphocytic leukemia; chronic myelogenous leukemia; and hairy cell leukemia); a lymphoma (e.g., AIDS-related lymphoma; cutaneous T-cell lymphoma; Hodgkin's lymphoma, including adult T-cell lymphoma); Human and pediatric Hodgkin's lymphoma and Hodgkin's lymphoma during pregnancy; non-Hodgkin's lymphoma, including adult and pediatric non-Hodgkin's lymphoma and non-Hodgkin's lymphoma during pregnancy; mycosis fungoides; Sézary syndrome; Waldenstrom's macroglobulinemia; primary mediastinal large B-cell lymphoma; mantle cell lymphoma; diffuse large B-cell lymphoma; and primary central nervous system lymphoma); and other hematologic cancers (e.g., chronic myeloproliferative disorders; multiple myeloma / plasma cell neoplasms; myelodysplastic syndromes; and myelodysplastic / myeloproliferative disorders); osteosarcoma; ovarian cancer; breast cancer; lung cancer; glioblastoma; retinoblastoma; and metastases of the above cancers.

45. Use of a pharmaceutical formulation according to claims 1 to 32, or a composition according to claims 33 to 35, or a kit according to claims 36 to 38 for the manufacture of a medicament for treating or preventing cancer, e.g., wherein the cancer is selected from the group consisting of: a blood cancer / blood cell cancer, e.g., a leukemia (e.g., acute lymphoblastic leukemia, including adult and childhood acute lymphoblastic leukemia; acute myeloid leukemia, including adult and childhood acute myeloid leukemia; chronic lymphocytic leukemia, such as B-cell chronic lymphocytic leukemia; chronic myelogenous leukemia; and hairy cell leukemia); a lymphoma (e.g., AIDS-related lymphoma; cutaneous T-cell lymphoma; Hodgkin's lymphoma); The present invention also includes hematologic malignancies, including Hodgkin's lymphoma in adults and children, and Hodgkin's lymphoma during pregnancy; non-Hodgkin's lymphoma in adults and children, and non-Hodgkin's lymphoma during pregnancy; mycosis fungoides; Sézary syndrome; Waldenstrom's macroglobulinemia; primary mediastinal large B-cell lymphoma; mantle cell lymphoma; diffuse large B-cell lymphoma; and primary central nervous system lymphoma; and other hematologic cancers (e.g., chronic myeloproliferative disorders; multiple myeloma / plasma cell neoplasms; myelodysplastic syndromes; and myelodysplastic / myeloproliferative disorders); osteosarcoma; ovarian cancer; breast cancer; lung cancer; glioblastoma; retinoblastoma; and metastases of the above cancers.

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