Bis-chloroethylamino derivatives for treating leukemia

Abstract

The invention provides compounds of formula (I), that are peptidase enhanced cytotoxics for use for use in the treatment of and / or prophylaxis of BCL2 inhibitor resistant or refractory acute myeloid leukaemia (AML). (Formula)

Description

[0001] Novel Therapeutic Treatments

[0002] Field of the Invention

[0003] The present invention relates to novel therapeutic treatments, particularly for use in the field of cancer. More particularly, but not exclusively, this invention concerns compounds and compositions thereof for use in the treatment and / or prophylaxis of haematological cancers. In particular, the compounds and compositions thereof are for use in the treatment and / or prophylaxis of acute myeloid leukaemia (AML) and more particularly in the treatment of BCL2 inhibitor-resistant AML.

[0004] Background of the Invention

[0005] An increased expression of various hydrolytic enzymes like peptidases, esterases, and proteases has been described in several types of human malignancies, especially those characterized by fast-growing and aggressive phenotypes (Pharmacol Ther. 1999; 83: 67-123). The Zn2_,+-dependent membrane-bound ectopeptidase aminopeptidase N (APN, also known as CD 13), widely expressed in mammalian cells, plays an important role in the development of cancer, including processes like tumour cell invasion, differentiation, proliferation, apoptosis, motility, and angiogenesis (Cancer Lett. 2006; 243: 135-43; Curr Med Chem. 2007; 14: 639-47; Curr Med Chem. 2008; 15: 2850-65). The multiple functions of APN have led to its designation as a “moonlighting ectoenzyme” (Trends Mol Med. 2008; 14: 361-71). Together, these abilities suggest APN as a potential therapeutic target in the treatment and / or prophylaxis of cancer. Different approaches have been used to develop new drugs directed to this target, including enzyme inhibitors and APN-targeted carrier constructs (Cancer Sci. 2011; 102: 501-8).

[0006] One class of drug that may find use in the treatment and / or prophylaxis of cancers by exploitation of hydrolytic enzymes, such as peptidases (e.g. APN), esterases, and proteases, which are present in malignancies, is peptide drug conjugates (PDCs). PDCs such as melflufen and the new family of peptide drug conjugates (described in International Patent Application WO2022 / 263679) that has recently been identified, find utility in the treatment and / or prophylaxis of cancers. In this regard, the PDCs use the increased peptidase and esterase expression to rapidly and selectively release cytotoxic agents inside the tumour cells (Wickstrbm, Malin et al. 2017; Miettinen, J. Juho et al. 2021).

[0007] Melflufen (also known as melphalan flufenamide) was approved by the European Medical Agency for relapsed / refractory multiple myeloma in August 2022. Its preparation was described in WOOl / 96367.

[0008] The current most common first line treatment for acute myeloid leukaemia, especially for older patients who are unfit for intensive chemotherapy, is the combination of the BCL2 inhibitor, Venetoclax, with a hypermethylating agent, decitabine or azacitabine or a low dose cytarabine. This has been especially effective in the induction of remission in older, previously untreated-AML patients. However, the treatment has a relatively short duration of response, and it suffers from the development of resistance (Saliba, John et al. 2021; Ong, Kim et al. 2022). BCL2 is also referred to as Bel -2.

[0009] Accordingly, despite the advances that have been made, there is still a need for improved treatments, particularly for patients with relapsed / refractory disease, for non-responders and in cases where the cancer develops resistance to existing therapies. In particular, there is an urgent need for new therapeutic treatment options especially for patients, resistant, relapsing or refractory to a BCL2 inhibitor-based treatment. Such patients may have been previously exposed to the BCL2 inhibitor. More particularly, there is a need for new therapeutic treatment options especially for patients who are resistant, relapsing or refractory to a venetoclax-based treatment. Such patients may have been previously exposed to venetoclax.

[0010] The present invention, therefore, seeks to meet the aforementioned needs.

[0011] In particular, the present invention seeks to provide new treatment options for patients who are resistant, relapsing or refractory to BCL2 inhibitor-based (e.g., venetoclax- based) treatment, in particular providing an improved treatment for use in the treatment of and / or prophylaxis of BCL2 inhibitor resistant or refractory acute myeloid leukaemia (AML). Summary of the Invention

[0012] The present invention provides a compound according to formula (I), or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate, wherein,

[0013] X is Ci-6 alkylene;

[0014] Wi, W2, W3 and W4 are each CH, or one of Wi, W2, W3 and W4 is N and the others are CH;

[0015] R1is selected from the group consisting of H; Ci-4alkyl, optionally substituted by 1, 2 or 3 groups independently selected from halogen; and halogen;

[0016] R2is selected from the group consisting of H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and Ci-ealkyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and

[0017] R3is a group according to formula (II): or formula (III): U) wherein,

[0018] R4is selected from the group consisting of N(Rc)(Rd) and formula (IV): when R4is formula (IV), R5is Rb; and when R4is N(Rc)(Rd), R5is selected from the group consisting of Rband formula (V): wherein each Rais independently selected from the group consisting of H;

[0019] Ci-ealkyl; -CH2-phenyl; or -CH2-3 to 12-membered heterocyclyl comprising 1, 2, 3 or 4 heteroatoms selected from N, O and S; wherein said Ci-ealkyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of -OH, -OCi-6alkyl, -NH2, -NHC(=NH)NH2, -C(O)OH, -C(O)NH2, -SH, -SCH3, and halogen; and said phenyl or heterocyclyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen, -NH2, -OH, -OCi-ealkyl and -NO2;

[0020] Rbis selected from the group consisting of -OH ; -N(Re)(Rf); and -OCi-ealkyl optionally substituted by one or more groups selected from halogen, -OH, -CN, - N(Re)(Rf), -Ce-ioaryl, or 3 to 12 membered heterocycle comprising one or more O, N or S atoms and optionally substituted by 1, 2 or 3 halogens, and / or wherein said alkyl is optionally interrupted by 1, 2 or 3 O, N or S atoms; Rcand Rdare each independently selected from the group consisting of H; -Ci-ealkyl, C(O)Ci-ealkyl; and -CFL-phenyl, wherein said alkyl or said phenyl is optionally substituted by 1, 2 or 3 groups selected from halogen; and

[0021] Reand Rfare each independently selected from the group consisting of H and - Ci-ealkyl, wherein said alkyl is optionally substituted by 1, 2 or 3 groups selected from halogen; or Reand Rftogether with the nitrogen atom to which they are attached form a 4-, 5- or 6-membered heterocycle which is optionally substituted by 1, 2 or 3 groups selected from halogen, for use in the treatment of and / or prophylaxis of BCL2 inhibitor resistant or refractory acute myeloid leukaemia (AML).

[0022] The present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, as defined herein, for use in the treatment of and / or prophylaxis of venetoclax -resistant amyloid myeloid leukaemia (AML).

[0023] The present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, as defined herein, for use in the treatment of and / or prophylaxis of BCL2 inhibitor-resistant AML wherein there has been previous exposure to a BCL2 inhibitor. In this regard, the invention provides a composition comprising a compound of formula (I), as defined herein, for use in the treatment of and / or prophylaxis of BCL2 inhibitor-resistant AML in a subject who has previously received treatment with a BCL2 inhibitor. For example, the compound or composition of the invention may be administered to a subject who has received a BCL2 inhibitor for the treatment of AML, wherein the BCL2 inhibitor was ineffective at treating the AML or the AML returned once treated with the BC12 inhibitor was ceased.

[0024] The compound for use according to the present invention described herein, wherein the compound of formula (I) has the structure (la):

[0025]

[0026] Preferably, the compound for use according to the present invention as described herein wherein R1is H.

[0027] More preferably, the compound for use according to the present invention as described herein wherein R2is selected from the group consisting of phenyl optionally substituted by 1, 2 or 3 halogens; and -Ci-ealkyl optionally substituted by 1, 2 or 3 halogens; for example -Ci-4alkyl optionally substituted by 1, 2 or 3 halogens, for example methyl.

[0028] Preferably, the compound for use according to the present invention as described herein wherein X is -CH2- or -CH2-CH2-.

[0029] The compound for use according to the present invention as described herein wherein R3is a group according to formula (Ila): or formula (Illa): Preferably, the compound for use according to the present invention as described herein wherein R4is selected from the group consisting of N(Rc)(Rd) and formula (IVa): According to a preferred embodiment, the compound for use according to the present invention as described herein wherein, when R4is formula (IV) or (IVa), R5is Rb; and when R4is N(Rc)(Rd), R5is selected from the group consisting of Rband formula (Va), wherein formula (Va) has the following structure,

[0030] The compound for use according to the present invention as described herein wherein R4is NH2. Preferably, the compound for use according to the present invention as described herein wherein each Rais independently selected from the group consisting of Ci-ealkyl and -CH2-phenyl; wherein said Ci-ealkyl is optionally substituted by -OH; -NH2; -NHC(=NH)NH2;

[0031] -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen (for example, F or Cl); and said phenyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen (for example, F or Cl); -NH2; -OH; -O-Ci-ealkyl; and - NO2.

[0032] Preferably, the compound for use according to the present invention as described herein wherein each Rais selected from the group consisting of -Ci-ealkyl and -CH2- phenyl; wherein said phenyl is optionally substituted by 1, 2 or 3 halogen substituents.

[0033] The compound for use according to the present invention as described herein wherein when R4is formula (IV) or (IVa), R5is -OCi-ealkyl; and when R4is NH2, R5is -OCi- ealkyl or formula (V) or (Va).

[0034] More preferably, the compound for use according to the present invention as described herein wherein when Rbis -OCi-ealkyl, it is selected from the group consisting of methoxy, ethoxy and isopropoxy. Preferably, the compound of formula (I) for use in the treatment of and / or prophylaxis of BCL2 inhibitor-resistant acute myeloid leukaemia (AML) (for example, venetoclax-resistant AML) is selected from: Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 1);

[0035] Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-3-(4-fluorophenyl)propanoate (Example compound 2);

[0036] Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-3-methyl-butanoate (Example compound 3);

[0037] Ethyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]propanoyl]amino]-4-methyl-pentanoate (Example compound 4);

[0038] Ethyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]propanoyl]amino]-3-(4-fluorophenyl)propanoate (Example compound 5); Ethyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]propanoyl]amino]-3-methyl-butanoate (Example compound 6);

[0039] Ethyl (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-4-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]butanoate (Example compound 7);

[0040] Ethyl (2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propanoyl]amino]-4-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]butanoate (Example compound 8);

[0041] Ethyl (2S)-2-[[(2S)-2-amino-3-methyl-butanoyl]amino]-4-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]butanoate (Example compound 9);

[0042] Ethyl (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-3-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]propanoate (Example compound 10);

[0043] Ethyl (2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propanoyl]amino]-3-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]propanoate (Example compound 11);

[0044] Ethyl (2S)-2-[[(2S)-2-amino-3-methyl-butanoyl]amino]-3-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]propanoate (Example compound 12);

[0045] Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-phenyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 13);

[0046] Methyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]propanoyl]amino]-4-methyl-pentanoate (Example compound 14);

[0047] Isopropyl (2 S)-2- [ [(2 S)-2-amino-3 - [5 - [bi s(2-chloroethyl)amino] - 1 -methyl- benzimidazol-2-yl]propanoyl]amino]-4-methyl-pentanoate (Example compound 15);

[0048] Ethyl (25)-2-[[(25)-2-[[(25)-2-amino-3-(4-fluorophenyl)propanoyl]amino]-4-methyl- pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoate (Example compound 16);

[0049] Ethyl (2S)-2-[[(2R)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propanoyl]amino]-4-methyl- pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoate (Example compound 21)

[0050] Ethyl (2S)-2-[[(2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoyl]amino]-3-(4- fluorophenyl)propanoate (Example compound 22)

[0051] Ethyl (2S)-2-[[(2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoyl]amino]-4-methyl-pentanoate (Example compound 23)

[0052] 2 -Morpholinoethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 24) 2-Isopropoxyethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 25) Isopropyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 26) Methyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 27)

[0053] 3-(Dimethylamino)propyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l- methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 28)

[0054] (2-Methoxy-l-methyl-ethyl) (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l- methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 29)

[0055] (2S)-2-[[(2S)-2-Amino-4-[5-[bis(2-chloroethyl)amino]-l-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]-l-methyl-benzimidazol-2- yl]butanoyl]amino]-N,4-dimethyl-pentanamide (Example compound 31)

[0056] Ethyl (2S)-2-[[(2S)-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2-yl]-2- (methylamino)butanoyl]amino]-4-methyl-pentanoate (Example compound 32) Ethyl (2S)-2-[[(2S)-2-acetamido-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-3-(4-fluorophenyl)propanoate (Example compound 34)

[0057] Ethyl (2S)-2-[[(2S)-2-acetamido-3-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]propanoyl]amino]-4-methyl-pentanoate (Example compound 35) Ethyl (2S)-2-[[(2S)-2-acetamido-3-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]propanoyl]amino]-3-(4-fluorophenyl)propanoate (Example compound 36)

[0058] Ethyl (2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoate (Example compound 38)

[0059] Ethyl (2R)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 45)

[0060] Ethyl (2S)-2-[[(2R)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 46) Ethyl (2R)-2-[[(2R)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 47)

[0061] Ethyl (2S)-2-[[(2S)-2-amino-4-[6-[bis(2-chloroethyl)amino]-3-methyl-imidazo[4,5-b] pyridin-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 48); and Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloro-l,l,2,2-tetradeuterio-ethyl)amino]-l- methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 50) or a pharmaceutically acceptable salt, amide or carbamate thereof, including a salt of such an amide or carbamate.

[0062] More preferably, the compound of formula (I) for use according to claim 1, wherein the compound is selected from the group consisting of:

[0063] (25)-2-[[(25)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoyl]amino]-3-methyl-butanoic acid (Example compound 19); and (25)-2-[[(25)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoyl]amino]-4-methyl-pentanoic acid (Example compound 20);

[0064] (2S)-2-[[(2S)-4-[5-[bis(2-chloroethyl)amino]-l-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]-l- methyl-benzimidazol-2-yl]butanoic acid (Example compound 37)

[0065] (2S)-2-[[(2S)-2-Amino-4-[5-[bis(2-chloroethyl)amino]-l-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]-l-phenyl-benzimidazol-2- yl]butanoyl]amino]-4-methyl-pentanoic acid (Example compound 40) 2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]propanoyl]amino]-3-methyl-butanoic acid (Example compound 42) (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-3-[5-[bis(2-chloroethyl)amino]-l- methyl-benzimidazol-2-yl]propanoic acid (Example compound 43)

[0066] (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]propanoyl]amino]-4-methyl-pentanoic acid (Example compound 44); and (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chl oro-1, 1,2, 2-tetradeuterio-ethyl)amino]- 1- methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (Example compound 51) or a pharmaceutically acceptable salt, amide or carbamate thereof, including a salt of such an amide or carbamate.

[0067] It has been found that compounds of formula (I), as defined above, each display surprisingly strong cytotoxicity in cells from patients with BCL2 inhibitor- resistant acute myeloid leukemia and, more particularly, in cells from patients from venetoclax-resistant acute myeloid leukemia (AML). Compounds of formula (I) are readily hydrolysed by enzymes within cancer cells to form metabolites that are preferentially sequestered and retained within cells and have strong alkylating activity.

[0068] According to an embodiment of the invention, there is provided a metabolite derived from a compound of formula (I) for use in the treatment of BCL2 inhibitor- resistant acute myeloid leukemia. Preferably, the metabolite is for use in the treatment of venetoclax-resistant AML. In certain embodiments, the compound or a composition according to the invention may comprise a metabolite of such a compound, wherein said metabolite has a structure according to formula (I), (la) or (lb), as described herein.

[0069] Preferably, the metabolite is: (25)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2-yl]propanoic acid (Example compound 17);

[0070] (5)-2-amino-4-(5-(bis(2-chloroethyl)amino)-l-methyl-lH-benzo[d]imidazol-2- yl)butanoic acid (Example compound 18);

[0071] Ethyl (2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoate (Example compound 38) (2S)-2-Amino-4-[5-[bis(2-chloroethyl)amino]-l-phenyl-benzimidazol-2-yl]butanoic acid (Example compound 41); or (2S)-2-amino-4-[5-[bis(2-chloro-l,l,2,2-tetradeuterio-ethyl)amino]-l-methyl- benzimidazol-2-yl]butanoic acid (Example compound 49); or a pharmaceutically acceptable salt, amide or carbamate thereof, including a salt of such an amide or carbamate. The invention also provides a composition comprising a compound of formula (I), as defined above, for use in the treatment of and / or prophylaxis of BCL2 inhibitor resistant, refractory acute myeloid leukaemia (AML). The composition is preferably a pharmaceutical composition.

[0072] The invention also provides a composition comprising a compound of formula (I), as defined herein, for use in the treatment and / or prophylaxis of venetoclax -resistant AML.

[0073] The invention provides a composition comprising a compound of formula (I), as defined herein, for use in the treatment of and / or prophylaxis of BCL2 inhibitorresistant AML wherein there has been previous exposure to the BCL2 inhibitor. In this regard, the invention provides a composition comprising a compound of formula (I), as defined herein, for use in the treatment of and / or prophylaxis of BCL2 inhibitor-resistant AML in a subject who has previously received treatment with a BCL2 inhibitor. For example, the compound or composition of the invention may be administered to a subject who has received a BCL2 inhibitor for the treatment of AML, wherein the BCL2 inhibitor was ineffective at treating the AML or the AML returned once treated with the BC12 inhibitor was ceased.

[0074] The invention provides a composition comprising a compound of formula (I), as defined above, for use in the treatment and / or prophylaxis of BCL2 inhibitor-resistant AML whereby an apoptotic agent (e.g. a BCL2 inhibitor) is given in combination with a compound of Formula (I).

[0075] The invention provides a composition comprising a compound of formula (I), as defined above, for use in the treatment and / or prophylaxis of venetoclax-resistant AML whereby an apoptotic agent (e.g. a BCL2 inhibitor) is given in combination with a compound of Formula (I).

[0076] Preferably, the invention further comprises one or more pharmaceutically acceptable carriers. The invention further provides a method of treating or preventing BCL2 inhibitorresistant AML in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I), as defined above, or a composition thereof.

[0077] The invention further provides a method of treating or preventing venetoclax-resistant AML in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I), as defined above, or a composition thereof.

[0078] The invention further provides a method of treating or preventing BCL2 inhibitor - resistant AML in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I), as defined above, or a composition thereof in combination with an apoptotic agent (e.g. a BCL2 inhibitor).

[0079] The invention further provides a method of treating or preventing venetoclax-resistant AML in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I), as defined above, or a composition thereof in combination with an apoptotic agent (e.g. a BCL2 inhibitor).

[0080] According to the present invention, there is provided a method of treating or preventing BCL2 inhibitor-resistant acute myeloid leukaemia in a subject in need thereof, the method comprising administering to the subject an effective amount of a metabolite compound derived from the compound as described herein.

[0081] Preferably, the method of treating or preventing venetoclax-resistant acute myeloid leukaemia in a subject in need thereof comprises administering to the subject an effective amount of a metabolite compound derived from the compound according to the present invention.

[0082] The invention further provides a compound of formula (I), as defined above, or a composition thereof for use in the manufacture of a medicament for the treatment and / or prophylaxis of BCL2 inhibitor-resistant AML. Preferably, the composition according to the present invention is for use in the manufacture of a medicament for the treatment and / or prophylaxis of venetoclax- resistant acute myeloid leukaemia.

[0083] The invention further provides a compound of formula (I) or a composition thereof as disclosed herein for use in the manufacture of a medicament for the treatment and / or prophylaxis of venetoclax-resistant AML, whereby the compound is given in combination with an apoptotic agent (e.g. a BCL2 inhibitor).

[0084] The apoptotic agent (e.g. a BCL2 inhibitor) which is given in combination with the compound of formula (I) is preferably venetoclax.

[0085] According to another embodiment of the present invention, there is provided a metabolite compound according to the present invention for use in the manufacture of a medicament for the treatment and / or prophylaxis of BCL2 inhibitor-resistant acute myeloid leukaemia.

[0086] Preferably, the metabolite compound according to the present invention is for use in the manufacture of a medicament for the treatment and / or prophylaxis of venetoclax- resistant acute myeloid leukaemia.

[0087] The compound or composition for use according to the present invention may further comprise one or more additional therapeutic agents, for example a therapeutic agent that is selected from the group consisting of: a steroid, a checkpoint inhibitor, a nuclear transport inhibitor, a BCL2 inhibitor or an apoptotic inhibitor, an adoptive cell therapy, a bi-specific T-cell engager, an immunomodulatory imide drug, a proteasome inhibitor, a histone deacetylase inhibitor, a tubulin inhibitor, a peptide drug conjugate, an alkylator, a topoisomerase II inhibitor and a DNA intercalator.

[0088] The invention also provides a kit comprising a compound of formula (I), as described herein or a composition thereof. Preferably, the kit further comprises one or more additional therapeutic agents. According to an embodiment of the invention, there is provided a method of selecting a BCL2 inhibitor-resistant patient to be treated with the compound of formula (I), as defined herein, or a composition thereof wherein the proportion of hematopoietic stem cells (HSC) / progenitor like cells and neutrophils determine whether a patient may be treated, wherein the patient sample tested has significantly lower fractions of HSC / progenitor like cells and higher proportions of HSC / multipotent progenitors (MPP) cells and neutrophils, thereby indicates sensitivity to the compound of formula (I). Preferably, the BCL2 inhibitor resistant patient is resistant to venetoclax.

[0089] According to the invention, the HSC / Progenitor like cells are present in the range 14 and 63% where there is high sensitivity to the compound of formula (I), as defined herein. Typically, when the HSC / Progenitor like cells in patient sample are present in the range of 14% and 63%, the patient exhibits high sensitivity to the compound of formula (I), as defined herein. For the avoidance of doubt, references herein to the percentage of specific cell types in a patient sample (e.g., the percentage of HSC / Progenitor like cells) refer to the proportion of that specific cell type relative to the total number of cells in the patient sample.

[0090] According to another embodiment of the invention, the HSC / Progenitor like cells are present in the range 14% and 63%, the HSC / MPP cells are present in the range 0 to 7% and the neutrophils are present in the range of 2% to 14% where there is high sensitivity to the compound of formula (I), as defined herein.

[0091] Preferably, the presence of HSC / Progenitor like cells is about 24%, where there is high sensitivity to the compound of formula (I), as defined herein.

[0092] Preferably, the presence of HSC / Progenitor like cells is about 24%, the presence of HSC / MPP cells is about 4% and the presence of neutrophils is about 7% where there is high sensitivity to the compound of formula (I), as defined herein.

[0093] According to the invention, there is provided a method of selecting a patient as described above, wherein the HSC / Progenitor like cells are present in the range 15% and 63%, the HSC / MPP cells are present in the range 0% to 3.5% and the neutrophils are present in the range of 0% to 7.5% where there is low sensitivity to the compound of formula (I).

[0094] According to yet another embodiment of the invention, there is a method of selecting a patient as described above, wherein the ratio between HSC / Progenitor like cells divided by neutrophils is lower than 50%. Preferably, the ratio between HSC / Progenitor like cells divided by neutrophils is lower than 30%. More preferably, the ratio between HSC / Progenitor like cells divided by neutrophils is lower than 10%.

[0095] According to a more preferred method of selecting a patient according to the invention, the ratio between HSC / Progenitor like cells divided by neutrophils is lower than 3.6.

[0096] Description of the Figures

[0097] Figure 1 is a heat map indicating the drug sensitivity scores (DSS) of venetoclax, melflufen and Example compound 1 in a blast cell population for 20 AML samples. The heat map shows the differences between disease stage, gender, age, FAB-type and malignant cell numbers.

[0098] Figure 2 shows DSS scores for Example compound 1 and melflufen when compared to venetoclax.

[0099] Figure 3 shows the mean DSS scores for both the parental and venetoclax resistant (VenR) cell lines, HL-60, Kasumi-1, MOLM-13 and MV4-11.

[0100] Figure 4 shows a single cell RNA-sequencing analysis of samples. A UMAP projection of aggregated cells from 20 AML samples wherein each dot represents the transcriptome of a single cell and colour code defining each cluster.

[0101] Figure 5 is a boxplot depicting the percentage of different cell-type populations predicted in 20 AML patient delivered samples where HSC / Prog-like cells were the most enriched cell-type. Figure 5 shows the cell subtype % analysis of 17 different cell-type populations that were identified, including different myeloid and lymphoid cell-types. Figure 6 shows Cell-types (n+3) with the significant (P< 0.01) difference in the cell % in the less-sensitive and in the highly-sensitive sub-groups to Example compound of Example 1.

[0102] Figure 7 shows scatter plots showing the correlation of Example compound 1 drug sensitivity scores in the blast cell population with cell percentages of (A) HSC / Prog like cell, (B) HSC / MPP cells and (C) Neutrophils.

[0103] Detailed Description

[0104] As discussed above, it has been found that compounds of formula (I), as defined above, each display surprisingly strong cytotoxicity in cells from patients with BCL2 inhibitor-resistant acute myeloid leukemia (AML).

[0105] The invention, therefore, provides a compound of formula (I), as defined above, or a composition thereof for use in the treatment and / or prophylaxis of BCL2 inhibitorresistant AML. It has also surprisingly been found that the inhibition by the compounds of formula (I) as defined herein, is especially notable when the compound is used in the treatment of and / or prophylaxis of venetoclax-resistant acute myeloid leukaemia.

[0106] The compounds of formula (I), as defined herein, are a new class of PDC compounds that are highly cytotoxic towards human cancer cells that have recently been identified, and the compounds are particularly cytotoxic towards human haematological cancer cells. As described in PCT patent application number PCTZEP2022 / 066756 (which was published under the number WO2022 / 263679), various examples of such compounds have been synthesised. The compounds were found to be highly potent and display selectivity towards cancer cells. In addition, the compounds were found to be readily hydrolysed within the cancer cells to form metabolites that were sequestered and retained within the cells and had strong alkylating activity, thus demonstrating that the compounds as described herein are an effective new class of PDC for the treatment and / or prophylaxis of cancer, and particularly the treatment and / or prophylaxis of haematological cancers. Accordingly, the present invention provides a compound of formula (I): wherein, Wi, W2, W3, W4, R1, R2, R3and X are as defined herein, for use in the treatment of and / or prophylaxis of BCL2 inhibitor resistant or refractory acute myeloid leukaemia (AML).

[0107] Also provided is a compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate for use in the treatment of and / or prophylaxis of BCL2 inhibitor-resistant AML wherein there has been previous exposure to the BCL2 inhibitor. In this regard, the invention provides a composition comprising a compound of formula (I), as defined herein, for use in the treatment of and / or prophylaxis of BCL2 inhibitorresistant AML in a subject who has previously received treatment with a BCL2 inhibitor. For example, the compound or composition of the invention may be administered to a subject who has received a BCL2 inhibitor for the treatment of AML, wherein the BCL2 inhibitor was ineffective at treating the AML or the AML returned once treated with the BC12 inhibitor was ceased.

[0108] In an embodiment, the compound of formula (I), as defined herein, is for use in the treatment of and / or prophylaxis of venetoclax -resistant acute myeloid leukaemia.

[0109] In an embodiment, W2, W3 and W4 are each CH, and Wi is N. It is readily seen that the -N(CH2CH2C1)2 group is attached to the core part of the molecule through a carbon atom on the ring. For the relevant W group, the H in the CH group is correspondingly absent.

[0110] In preferred embodiments, Wi, W2, W3 and W4 are each CH and so formula (I) has the formula (la):

[0111] (la).

[0112] In preferred embodiments, formula (I) is according to formula (lb),

[0113] In the compounds of formula (I), X is Ci-ealkylene. For example, X may be Ci- 4alkylene, Ci-3alkylene, Ci-2alkylene, C2alkylene or Cialkylene. X may be a linear or branched alkylene. The alkylene at position X forms a link between the imidazole moiety and the peptide portion of the compound (i.e. the R3portion of the compound, as described below). Alkylene linkers at the X position that are one carbon or two carbons in length have been found to be particularly effective. As such, in preferred embodiments, X is a Cialkylene (i.e. -CH2-) or linear C2alkylene (i.e. -CH2-CH2-).

[0114] In the compounds of formula (I), R1is selected from the group consisting of H; Ci- 4alkyl, optionally substituted by 1, 2 or 3 groups independently selected from halogen; and halogen (for example selected from the group consisting of H; Ci-4alkyl; and halogen.). For example, R1may be selected from the group consisting of H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, F, Cl, Br and I. Optionally, those groups may be substituted by 1, 2 or 3 groups independently selected from halogen. In certain embodiments, R1is selected from the group consisting of H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, F and Cl. In certain preferred embodiments, R1is H.

[0115] In the compounds of formula (I), R2is selected from the group consisting of H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and - Ci-ealkyl optionally substituted by 1, 2 or 3 groups independently selected from halogen. For example, R2may be selected from the group consisting of H; phenyl optionally substituted by 1, 2 or 3 F or Cl; and -Ci-4alkyl (for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert-butyl) optionally substituted by 1, 2 or 3 F or Cl. In an embodiment, R2is selected from the group consisting of phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and - Ci-ealkyl optionally substituted by 1, 2 or 3 groups independently selected from halogen, for example -Ci-4alkyl (for example methyl) optionally substituted by 1, 2 or 3 groups selected from halogen. In certain embodiments, R2is an unsubstituted -Ci- 2alkyl (for example, methyl) or an unsubstituted phenyl.

[0116] In the compounds of formula (I), R3is a group according to formula (II): or formula (III): In preferred embodiments, R3is a group according to formula (Ila): or formula (Illa): For the avoidance of doubt, in formulas (II), (III), (Ila) and (Illa) denotes the point of attachment of formula (II), (III), (Ila) or (Illa) to formula (I), (la) or (Ila).

[0117] In formulas (II), (Ila), (III) and (Illa), R4is selected from the group consisting of N(Rc)(Rd) and formula (IV):

[0118] In preferred embodiments, R4is selected from the group consisting of N(Rc)(Rd) and formula (IVa): For the avoidance of doubt, in formula (IV) and (IVa) denotes the point of attachment of formula (IV) or (IVa) to formula (II), (III), (Ila) or (Illa).

[0119] In compounds of formula (I), when R4is formula (IV) or (IVa), R5is Rb. Rbis selected from the group consisting of -OH; -N(Re)(Rf); and -OCi-ealkyl optionally substituted by one or more groups selected from halogen, -OH, -CN, - N(Re)(Rf), -Ce-ioaryl, or a 3 to 12 membered heterocycle comprising one or more O, N or S atoms and optionally substituted by 1, 2 or 3 halogens, and / or wherein said alkyl is optionally interrupted by 1, 2 or 3 O, N or S atoms.

[0120] For example, when R4is formula (IV) or (IVa), R5(which is Rb) may be selected from the group consisting of -OH and -OCi-4alkyl (for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, or tert-butoxy). Preferably, when R4is formula (IV) or (IVa), R5is selected from the group consisting of -OH, and -OCi- salkyl (for example, methoxy, ethoxy, propoxy, or isopropoxy). More preferably, when R4is formula (IV) or (IVa), R5is selected from the group consisting of methoxy, ethoxy or isopropoxy.

[0121] In compounds of formula (la), when R4is N(Rc)(Rd), R5is selected from the group consisting of Rband formula (V):

[0122] In preferred embodiments, when R4is N(Rc)(Rd), R5is -OH, -OCi-ealkyl or formula (Va):

[0123] For the avoidance of doubt, in formula (V) and (Va) denotes the point of attachment of formula (V) or (Va) to formula (II), (III), (Ila) or (Illa).

[0124] In embodiments wherein R4is N(Rc)(Rd), R5(being Rb) may be selected from the group consisting of -OH, -OCi-4alkyl (for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy or tert-butoxy), or R5may be selected from formula (V) and formula (Va). Preferably, when R4is N(Rc)(Rd), R5is selected from the group consisting of -OH, -OCi-3alkyl (for example, methoxy, ethoxy, propoxy or isopropoxy) and formula (Va). More preferably, when R4is N(Rc)(Rd), R5is selected from the group consisting of methoxy, ethoxy or isopropoxy.

[0125] Compounds of formula (I), in which R4is N(Rc)(Rd), formula (IV) or (IVa), and R5(being Rb) is -OCi-3alkyl (for example, methoxy, ethoxy or isopropoxy), have been found to be especially cytotoxic towards cancer cells. Thus, in certain preferred embodiments, in the compounds of formula (I), R5is -OCi-3alkyl (for example, methoxy, ethoxy or isopropoxy). That is to say that, when R4is N(Rc)(Rd), formula (IV) or (IVa), R5is preferably -OCi-3alkyl (for example, methoxy, ethoxy or isopropoxy).

[0126] In certain embodiments, R5is -OH.

[0127] In compounds of formula (I), each Rais independently selected from the group consisting of H; Ci-ealkyl; -CH2-phenyl; or -CH2-3 to 12-membered heterocyclyl comprising 1, 2, 3 or 4 heteroatoms selected from N, O and S; wherein said Ci-ealkyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of -OH, -OCi-6alkyl, -NH2, -NHC(=NH)NH2, -C(O)OH, -C(O)NH2, -SH, -SCH3, and halogen; and said phenyl or heterocyclyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen, - NH2, -OH, -OCi-ealkyl and -NO2. In certain embodiments, each Rais independently selected from the group consisting of H; -Ci-ealkyl; -CH2-indolyl; -CH2-phenyl; and -CH2-5-membered heteroaryl comprising 1, 2, 3 or 4 N or S atoms; wherein Ci-ealkyl is optionally substituted by -OH; -NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen (for example, F, Cl, Br or I); and phenyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen (for example, F, Cl, Br or I); -NH2; -OH; -OCi-ealkyl; and -NO2. Preferably, each Rais independently selected from the group consisting of H; Ci-4alkyl; and -CH2-phenyl; wherein alkyl or phenyl is optionally substituted by 1 or 2 halogen (for example, F or Cl). For example, each Ramay independently be methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, -CH2-phenyl, -CH2-fluorophenyl, -CH2- chlorophenyl, -CH2-difluorophenyl, or -CH2-di chlorophenyl. In certain preferred embodiments, each Ramay independently be isopropyl, isobutyl, sec-butyl, -CH2- phenyl, or -CH2-fluorophenyl (i.e. 2-fluorobenzyl, 3 -fluorobenzyl or 4-fluorobenzyl). In compounds of formula (I), Rbis selected from the group consisting of -OH; -N(Re)(Rf); and -OCi-ealkyl optionally substituted by one or more groups selected from halogen, -OH, -CN, -N(Re)(Rf), -Ce-ioaryl, or a 3 to 12 membered heterocycle comprising one or more O, N or S atoms and optionally substituted by 1, 2 or 3 halogens, and / or wherein said alkyl is optionally interrupted by 1, 2 or 3 O, N or S atoms. In certain embodiments, Rbis -OH or -OCi-ealkyl, wherein said alkyl is optionally interrupted by 1, 2 or 3 O or N atoms. In certain further embodiments, Rbis -OH or -OCi-ealkyl, for example -OCi-ealkyl. Preferably, Rbis -OCi-4alkyl. For example, Rbmay be methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy or secbutoxy. In certain preferred embodiments, Rbis methoxy, ethoxy or isopropoxy. More preferably, Rbis ethoxy.

[0128] In certain embodiments, Rbis -OH.

[0129] Rcand Rdare each independently selected from the group consisting of H, -Ci-ealkyl, -C(O)Ci-6alkyl, and -CH2-phenyl, wherein said alkyl or said phenyl is optionally substituted by 1, 2 or 3 groups selected from halogen. In preferred compounds, Rcis H and Rdis selected from H, -Ci-ealkyl and -C(O)Ci-6alkyl, wherein said alkyl is optionally substituted by 1, 2 or 3 groups selected from halogen. For example, Rcis H and Rdis selected from H, -Ci-4alkyl and C(O)Ci-4alkyl. For example, Rcis H and Rdis 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, Rcis H and Rdis selected from H, methyl and - C(O)methyl. Preferably, Rcis H and Rdis H.

[0130] In compounds of formula (I), Reand Rfare each independently selected from the group consisting of H and -Ci-ealkyl, wherein said alkyl is optionally substituted by 1, 2 or 3 groups selected from halogen; or Reand Rftogether with the nitrogen atom to which they are attached form a 4-, 5- or 6-membered heterocycle which is optionally substituted by 1, 2 or 3 groups selected from halogen. In preferred compounds, Reand Rfare each independently selected from the group consisting of H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl and sec-butyl. For example, Reand Rfare each independently selected from the group consisting of H and methyl. Preferably, Reand Rfare both H, both methyl, or one of Reand Rfis H and the other is methyl. In certain embodiments, the compounds are according to formula (la), wherein,

[0131] X is Ci-2alkylene (i.e. -CH2- or -CH2-CH2-);

[0132] R1is H;

[0133] R2is -Ci-2alkyl optionally substituted by 1, 2 or 3 F, or phenyl optionally substituted by 1, 2 or 3 F, for example, R2may be an unsubstituted -Ci-2alkyl or an unsubstituted phenyl;

[0134] R3is formula (Ila) or (Illa);

[0135] R4is selected from the group consisting of N(Rc)(Rd) and formula (IVa); when R4is formula (IVa), R5(being Rb) is -OCi-4alkyl; and when R4is N(Rc)(Rd), R5is

[0136] -OCi-4alkyl or formula (Va); each Rais independently selected from the group consisting of H; -Ci-ealkyl;

[0137] -CFh-indolyl; -CFh-phenyl; or -CH2-5-membered heteroaryl comprising 1, 2, 3 or 4 N or S atoms; wherein said -Ci-ealkyl is optionally substituted by -OH; -NH2;

[0138] -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and said phenyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen; -NH2; -OH; -O-Ci-ealkyl; and -NO2.;

[0139] Rbis -OCi-ealkyl; and

[0140] Rcis H and Rdis selected from H, -Ci-4alkyl and -C(O)Ci-4alkyl; or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate.

[0141] In certain other embodiments, the compound is according to formula (la), wherein,

[0142] X is Ci-2alkylene (i.e. -CH2- or -CH2-CH2-);

[0143] R1is H;

[0144] R2is methyl or phenyl;

[0145] R3is formula (Ila) or (Illa);

[0146] R4is N(Rc)(Rd) or formula (IVa);

[0147] R5(being Rb) is -OCi-salkyl; and each Rais independently selected from the group consisting of H; -Ci-ealkyl; -CH2- indolyl; -CH2-phenyl; or -CH2-5-membered heteroaryl comprising 1, 2, 3 or 4 N or S atoms; wherein Ci-ealkyl is optionally substituted by -OH; -NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and phenyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen;

[0148] -NH2; -OH; -OCi-6alkyl; and -NO2; or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate.

[0149] It has been found that compounds of formula (I) as described hereinabove are especially cytotoxic towards haematological cancer cells in an in vitro cytotoxicity assay and in ovo chicken embryo xenograft model of lymphoma, when the compound is according to formula (la), wherein,

[0150] X is Ci-2alkylene (i.e. -CH2- or -CH2-CH2-);

[0151] R1is H;

[0152] R2is methyl or phenyl;

[0153] R3is formula (Ila) or (Illa);

[0154] R4is N(Rc)(Rd) or formula (IVa);

[0155] R5(being Rb) is -OCi-3alkyl; and each Rais independently selected from the group consisting of isopropyl, isobutyl and -CH2-phenyl optionally substituted by 1 F (for example, 2-fluorobenzyl,

[0156] 3 -fluorobenzyl or 4-fluorobenzyl); or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate.

[0157] In certain other embodiments, the compounds of formula (I) as described hereinabove are according to formula (la), wherein,

[0158] X is Ci-2alkylene (i.e. -CH2- or -CH2-CH2-);

[0159] R1is H;

[0160] R2is -Ci-2alkyl optionally substituted by 1, 2 or 3 F, or phenyl optionally substituted by 1, 2 or 3 F, for example, R2may be an unsubstituted -Ci-2alkyl or an unsubstituted phenyl;

[0161] R3is formula (Ila);

[0162] R4is NH2;

[0163] R5(being Rb) is -OCi-4alkyl; and each Rais independently selected from the group consisting of H; -Ci-ealkyl; -CH2- indolyl; -CH2-phenyl; or -CH2-5-membered heteroaryl comprising 1, 2, 3 or 4 N or S atoms; wherein Ci-ealkyl is optionally substituted by -OH; -NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and phenyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen; -NH2; -OH; -O-Ci-6alkyl; and -NO2; or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate.

[0164] In certain other embodiments, the compounds of formula (I) as described hereinabove are according to formula (la), wherein,

[0165] X is -Ci-2alkylene (i.e. -CH2- or -CH2-CH2-);

[0166] R1is H;

[0167] R2is -Ci-2alkyl optionally substituted by 1, 2 or 3 F, or phenyl optionally substituted by 1, 2 or 3 F, for example, R2may be an unsubstituted -Ci-2alkyl or an unsubstituted phenyl;

[0168] R3is formula (Illa);

[0169] R4is NH2;

[0170] R5is -OCi-4alkyl; and each Rais independently selected from the group consisting of H; -Ci-ealkyl; -CH2- indolyl; -CH2-phenyl; or -CH2-5-membered heteroaryl comprising 1, 2, 3 or 4 N or S atoms; wherein Ci-ealkyl is optionally substituted by -OH; -NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and phenyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen;

[0171] -NH2; -OH; -O-Ci-6alkyl; and -NO2; or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate.

[0172] In certain other embodiments, the compounds of formula (I) as described hereinabove are according to formula (la), wherein

[0173] X is Ci-2alkylene (i.e. -CH2- or -CH2-CH2-);

[0174] R1is H;

[0175] R2is -Ci-2alkyl optionally substituted by 1, 2 or 3 F, or phenyl optionally substituted by 1, 2 or 3 F, for example, R2may be an unsubstituted -Ci-2alkyl or an unsubstituted phenyl;

[0176] R3is formula (Illa); R4is formula (IVa);

[0177] R5is -OCi-4alkyl; and each Rais independently selected from the group consisting of H; -Ci-ealkyl; -CH2- indolyl; -CH2-phenyl; or -CH2-5-membered heteroaryl comprising 1, 2, 3 or 4 N or S atoms; wherein -Ci-ealkyl is optionally substituted by -OH; -NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and phenyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen;

[0178] -NH2; -OH; -O-Ci-6alkyl; and -NO2; or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate.

[0179] The following definitions apply to the terms as used throughout this specification, unless otherwise limited in specific instances.

[0180] As used herein, the term "alkyl" means both straight and branched chain saturated hydrocarbon groups. Examples of alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, t-butyl, iso-butyl, sec-butyl, pentyl and hexyl groups. Among unbranched alkyl groups, there are preferred methyl, ethyl, n-propyl, iso-propyl and n- butyl groups. Among branched alkyl groups, there may be mentioned t-butyl, i-butyl, 1 -ethylpropyl and 1 -ethylbutyl groups.

[0181] As used herein, the term "cycloalkyl" means a saturated group in a ring system. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0182] As used herein, the term "halogen" means fluorine, chlorine, bromine or iodine. Fluorine, chlorine and bromine are preferred, and fluorine and chlorine are particularly preferred.

[0183] As used herein, the term “heteroaryl comprising 1, 2, 3 or 4 nitrogen, oxygen or sulphur atoms” means an aromatic cyclic group of carbon atoms wherein one, two, three or four of the carbon atoms is / are replaced by one, two, three or four heteroatoms independently selected from nitrogen, oxygen and sulphur (preferably nitrogen and sulphur). Examples of 5-membered heteroaryl comprising 1, 2 or 3 nitrogen and / or sulphur atoms include thiophene, thiazole, isothiazole, pyrrole, pyrroline, pyrazole, pyrazoline, imidazole, imidazoline, triazole and thiadiazole.

[0184] In certain preferred embodiments, the compound of formula (I) as described hereinabove is:

[0185] Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 1);

[0186] Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-3-(4-fluorophenyl)propanoate (Example compound 2);

[0187] Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-3-methyl-butanoate (Example compound 3);

[0188] Ethyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]propanoyl]amino]-4-methyl-pentanoate (Example compound 4);

[0189] Ethyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]propanoyl]amino]-3-(4-fluorophenyl)propanoate (Example compound 5);

[0190] Ethyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]propanoyl]amino]-3-methyl-butanoate (Example compound 6);

[0191] Ethyl (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-4-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]butanoate (Example compound 7);

[0192] Ethyl (2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propanoyl]amino]-4-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]butanoate (Example compound 8);

[0193] Ethyl (2S)-2-[[(2S)-2-amino-3-methyl-butanoyl]amino]-4-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]butanoate (Example compound 9);

[0194] Ethyl (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-3-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]propanoate (Example compound 10);

[0195] Ethyl (2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propanoyl]amino]-3-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]propanoate (Example compound 11);

[0196] Ethyl (2S)-2-[[(2S)-2-amino-3-methyl-butanoyl]amino]-3-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]propanoate (Example compound 12);

[0197] Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-phenyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 13);

[0198] Methyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]propanoyl]amino]-4-methyl-pentanoate (Example compound 14); Isopropyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]propanoyl]amino]-4-methyl-pentanoate (Example compound 15); Ethyl (25)-2-[[(25)-2-[[(25)-2-amino-3-(4-fluorophenyl)propanoyl]amino]-4-methyl- pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoate (Example compound 16);

[0199] Ethyl (2S)-2-[[(2R)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propanoyl]amino]-4-methyl- pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoate (Example compound 21)

[0200] Ethyl (2S)-2-[[(2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoyl]amino]-3-(4- fluorophenyl)propanoate (Example compound 22)

[0201] Ethyl (2S)-2-[[(2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoyl]amino]-4-methyl-pentanoate (Example compound 23)

[0202] 2 -Morpholinoethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 24)

[0203] 2-Isopropoxyethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 25) Isopropyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 26) Methyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 27)

[0204] 3-(Dimethylamino)propyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l- methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 28)

[0205] (2-Methoxy-l-methyl-ethyl) (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l- methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 29)

[0206] (2S)-2-[[(2S)-2-Amino-4-[5-[bis(2-chloroethyl)amino]-l-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]-l-methyl-benzimidazol-2- yl]butanoyl]amino]-N,4-dimethyl-pentanamide (Example compound 31)

[0207] Ethyl (2S)-2-[[(2S)-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2-yl]-2- (methylamino)butanoyl]amino]-4-methyl-pentanoate (Example compound 32) Ethyl (2S)-2-[[(2S)-2-acetamido-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-3-(4-fluorophenyl)propanoate (Example compound 34)

[0208] Ethyl (2S)-2-[[(2S)-2-acetamido-3-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]propanoyl]amino]-4-methyl-pentanoate (Example compound 35) Ethyl (2S)-2-[[(2S)-2-acetamido-3-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]propanoyl]amino]-3-(4-fluorophenyl)propanoate (Example compound 36)

[0209] Ethyl (2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoate (Example compound 38)

[0210] Ethyl (2R)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 45)

[0211] Ethyl (2S)-2-[[(2R)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 46)

[0212] Ethyl (2R)-2-[[(2R)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 47)

[0213] Ethyl (2S)-2-[[(2S)-2-amino-4-[6-[bis(2-chloroethyl)amino]-3-methyl-imidazo[4,5-b] pyridin-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 48); and Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloro-l,l,2,2-tetradeuterio-ethyl)amino]-l- methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 50) or a pharmaceutically acceptable salt, amide or carbamate thereof, including a salt of such an amide or carbamate.

[0214] The compounds of formula (I) as described hereinabove may be prepared using methods known to those skilled in the art of organic chemistry. Exemplary procedures for the preparation of compounds of formula (I) are described in WO2022 / 263679).

[0215] In embodiments, the compound of formula (I) as described hereinabove may comprise an isotope atom. As defined herein, an isotope 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 compound has a deuterium abundance level greater than the naturally occurring abundance of deuterium. The naturally occurring abundance of deuterium is 0.0156 mol%, wherein mol% is the percentage of the total moles of a sample’s hydrogen that is deuterium. Therefore, in 1 mole of naturally occurring hydrogen 0.156 mmol is deuterium, or in a sample of 6.022 x 1023naturally occurring hydrogen atoms there are 9.39 xlO19atoms of deuterium, or in a sample of 6413 naturally occurring hydrogen atoms there is one atom of deuterium. A deuterium abundance level greater than the naturally occurring abundance of deuterium may be at least 1 mol%, 5 mol%, 10 mol%, 50 mol%, 90 mol% or 98 mol% deuterium. In certain embodiments, the compound has a deuterium abundance level of 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, pg 29-40, and Hanson, The Organic Chemistry of Isotopic Labelling (2011), Chapter 3, RSC Publishing.

[0216] Depending upon the substituents present in compounds of formula (I) as described hereinabove, the compounds may form esters, amides, carbamates and / or salts. Salts of compounds of formula (I) as described hereinabove which are suitable for use in medicine are those wherein a counterion is pharmaceutically acceptable. However, salts having non-pharmaceutically acceptable counterions are within the scope of the present invention, for example, for use as intermediates in the preparation of the compounds of formula (I) as described hereinabove and their pharmaceutically acceptable salts, and physiologically functional derivatives. The term “physiologically functional derivative” refers to a chemical derivative of a compound of Formula (I) as described hereinabove that has the same physiological function as that compound, for example, by being convertible in the body thereto. Esters, amides and carbamates are examples of physiologically functional derivatives.

[0217] Salt forms suitable for use in the present invention include those formed with organic or inorganic acids or bases. In particular, suitable salts formed with acids include those formed with mineral acids, strong organic carboxylic acids, such as alkanecarboxylic acids of 1 to 4 carbon atoms which are unsubstituted or substituted, for example, by halogen, or such as saturated or unsaturated dicarboxylic acids, or such as hydroxycarboxylic acids, or such as amino acids, or with organic sulfonic acids, such as (Ci-C4)-alkyl- or aryl-sulfonic acids which are unsubstituted or substituted, for example by halogen. Pharmaceutically acceptable acid addition salts include those formed from hydrochloric, hydrobromic, sulphuric, nitric, citric, tartaric, acetic, phosphoric, lactic, pyruvic, acetic, trifluoroacetic, succinic, perchloric, fumaric, maleic, glycolic, lactic, salicylic, oxaloacetic, methanesulfonic, ethanesulfonic, p-toluenesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic, benzenesulfonic, isethionic, ascorbic, malic, phthalic, aspartic, and glutamic acids, lysine and arginine. Other acids such as oxalic, while not in themselves pharmaceutically acceptable, may be useful as intermediates in obtaining the compounds of formula (I) as described hereinabove and their pharmaceutical acceptable acid addition salts.

[0218] Pharmaceutically acceptable base salts include ammonium salts, alkali metal salts, for example those of potassium and sodium, alkaline earth metal salts, for example those of calcium and magnesium, and salts with organic bases, for example dicyclohexylamine, N-methyl-D-glucomine, morpholine, thiomorpholine, piperidine, pyrrolidine, a mono-, di- or tri-lower alkylamine, for example ethyl-, tert-butyl-, diethyl-, diisopropyl-, triethyl-, tributyl- or dimethyl-propylamine, or a mono-, di- or trihydroxy-lower alkylamine, for example mono-, di- or triethanolamine. Corresponding internal salts may furthermore be formed.

[0219] Preferred salts of a compound of formula (I) as described hereinabove as comprised in a composition of the present invention include acid addition salts such as those formed from hydrochloric, hydrobromic, acetic, p-toluenesulfonic, tartaric, sulphuric, succinic, phosphoric, oxalic, nitric, methanesulfonic, malic, maleic and citric acid. More preferably, the salt of a compound of formula (I) as described hereinabove is the hydrochloride salt (i.e. the addition salt formed from hydrochloric acid).

[0220] A compound which is itself inactive, but which, upon administration to the recipient, is capable of being converted into an active drug compound is known as a “prodrug”. A prodrug may, for example, be converted within the body, e.g. by hydrolysis in the blood, into an active form that has medical effects. Pharmaceutically acceptable prodrugs are described in T. Higuchi and V. Stella, Prodrugs as Novel Delivery Systems, Vol. 14 of the A. C. S. Symposium Series (1976); “Design of Prodrugs” ed. H. Bundgaard, Elsevier, 1985; and in Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987. The compounds of formula (I) as described hereinabove as comprised in a composition of the present invention can be provided in the form of a prodrug. Examples of prodrugs include esters, amides and carbamates.

[0221] Compounds of formula (I) as described hereinabove may have an appropriate group converted to an ester, an amide or a carbamate. Thus typical ester and amide groups formed from an acid group in a compound of Formula (I) as described hereinabove include -COORG, -CONRG2, -SCLOR*3, or -SO2N(RG)2, while typical ester and amide and carbamate groups formed from an -OH or -NHRGgroup in a compound of formula (I) as described hereinabove include -OC(O)RG, -NRGC(O)RG, -NRGCO2RG, -OSO2RG, and -NRGSO2RG, where RGis selected from the group consisting of Ci-salkyl, C2-salkenyl, C2-salkynyl, C3- scycloalkyl and Cs-scycloalkylCi-salkyl, haloC 1-8 alkyl, dihaloCi-salkyl, trihaloCi-salkyl, phenyl and phenylCi-4alkyl; more preferably RGis selected from the group consisting of Ci-ealkyl, C2-ealkenyl, C2-ealkynyl, C3-8cycloalkyl and C3- scycloalkylCi-ealkyl.

[0222] Those skilled in the art of organic chemistry will appreciate that many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as "solvates".

[0223] For example, a complex with water is known as a "hydrate". The complex may incorporate a solvent in stoichiometric or non-stoichiometric amounts. Solvates are described in Water-Insoluble Drug Formulation, 2ndedn, R. Lui, CRC Press, page 553, and Bym etal., Pharm. Res., 12(7), 1995, 945-954. Before it is made up in solution, a compound of Formula (I) as described hereinabove, as well as esters, amides, carbamates and / or salts thereof, may be in the form of a solvate. Solvates of a compound of Formula (I) as described hereinabove as comprised in a composition of the present invention that are suitable for use as a medicament are those wherein the associated solvent is pharmaceutically acceptable. For example, a hydrate is a pharmaceutically acceptable solvate. In the compositions, kits, methods and uses of the invention, the compound according to formula (I), or pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate can be in the form of a pharmaceutically acceptable solvate.

[0224] Metabolites

[0225] It has been found that compounds of Formula (I) as described hereinabove in which

[0226] R5is selected from the group consisting of -OCi-ealkyl, formula (V) and formula (Va) and Rbis a group selected from -N(Re)(Rf) and -OCi-ealkyl, are readily hydrolysed within cancer cells to form metabolites that are preferentially sequestered and retained within cells and have strong alkylating activity.

[0227] Thus, in certain embodiments, the compound for use according to the present invention is, or the composition for use according to the present invention comprises, a metabolite of such a compound, for example a metabolite having a structure according to formula (I), (la) or (lb), wherein,

[0228] X is Ci-6 alkylene;

[0229] Wi, W2, W3 and W4 are each CH, or one of Wi, W2, W3 and W4 is N and the others are CH; R1is selected from the group consisting of H; Ci-4 alkyl, optionally substituted by 1, 2 or 3 groups independently selected from halogen; and halogen;

[0230] R2is selected from the group consisting of H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and Ci-ealkyl optionally substituted by 1, 2 or 3 groups independently selected from halogen;

[0231] R3is a group according to formula (II), (III), (Ila) or (Illa);

[0232] R4is selected from the group consisting of N(Rc)(Rd), formula (IV) or (IVa); when R4is formula (IV) or (IVa), R5is -OH; and when R4is N(Rc)(Rd), R5is -OH, formula (V) or (Va); each Rais independently selected from the group consisting of H; -Ci-ealkyl; -CH2- phenyl; or -CH2-3 to 12-membered heterocyclyl comprising 1, 2, 3 or 4 heteroatoms selected from N, O and S; wherein said -Ci-ealkyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of -OH, -OCi-6alkyl, -NH2, -NHC(=NH)NH2, -C(O)OH, -C(O)NH2, -SH, -SCH3, and halogen; and said phenyl or heterocyclyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen, -NH2, -OH, -OCi-ealkyl and -NO2 and

[0233] Rbis OH; or a salt thereof.

[0234] Preferred radicals within the metabolite compounds as described immediately above, are as described hereinabove regarding compounds of Formula (I), (la) and (lb). For example, in certain embodiments, the metabolite has a structure according to formula (I), (la) or (lb), wherein,

[0235] X is -Ci-2alkylene (i.e. -CH2- or -CH2-CH2-);

[0236] R1is H;

[0237] R2is methyl or phenyl;

[0238] R3is formula (Ila) or (Illa);

[0239] R4is selected from the group consisting of N(Rc)(Rd) or formula (IVa); when R4is formula (IVa), R5is -OH; and when R4is N(Rc)(Rd), R5is -OH or formula (Va); each Rais independently selected from the group consisting of H; -Ci-ealkyl; -CH2- indolyl; -CH2-phenyl; or -CH2-5-membered heteroaryl comprising 1, 2, 3 or 4 N or S atoms; wherein -Ci-ealkyl is optionally substituted by -OH; -NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and phenyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen; -NH2; -OH; -O-Ci-ealkyl; and -NO2; and Rbis OH; or a salt thereof.

[0240] In certain embodiments, the metabolite has a structure according to formula (I), (la) or (lb), wherein,

[0241] X is Ci-2alkylene (i.e. -CH2- or -CH2-CH2-);

[0242] R1is H;

[0243] R2is methyl or phenyl;

[0244] R3is formula (Ila) or (Illa);

[0245] R4is NH2 or formula (IVa);

[0246] R5is -OH; each Rais independently selected from the group consisting of H; -Ci-ealkyl; -CH2- indolyl; -CH2-phenyl; or -CH2-5-membered heteroaryl comprising 1, 2, 3 or 4 N or S atoms; wherein Ci-ealkyl is optionally substituted by -OH; -NH2; -NHC(=NH)NH2; -C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen; and phenyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen;

[0247] -NH2; -OH; -OCi-ealkyl; and -NO2. Preferably, each Rais independently selected from the group consisting of isopropyl, isobutyl and -CH2-phenyl optionally substituted by 1 F (for example, 2-fluorobenzyl, 3-fluorobenzyl or 4-fluorobenzyl); or a salt thereof.

[0248] In certain preferred embodiments, the metabolite is: (25)-2-[[(25)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoyl]amino]-3-methyl-butanoic acid (Example compound 19); and (25)-2-[[(25)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoyl]amino]-4-methyl-pentanoic acid (Example compound 20);

[0249] (2S)-2-[[(2S)-4-[5-[bis(2-chloroethyl)amino]-l-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]-l- methyl-benzimidazol-2-yl]butanoic acid (Example compound 37) (2S)-2-[[(2S)-2-Amino-4-[5-[bis(2-chloroethyl)amino]-l-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]-l-phenyl-benzimidazol-2- yl]butanoyl]amino]-4-methyl-pentanoic acid (Example compound 40) 2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]propanoyl]amino]-3-methyl-butanoic acid (Example compound 42) (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-3-[5-[bis(2-chloroethyl)amino]-l- methyl-benzimidazol-2-yl]propanoic acid (Example compound 43) (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]propanoyl]amino]-4-methyl-pentanoic acid (Example compound 44); and (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chl oro-1,1, 2, 2-tetradeuterio-ethyl)amino]-l- methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (Example compound 51), or a salt thereof.

[0250] It has been further found that compounds of formula (I) as described hereinabove are readily further hydrolysed within cancer cells to form further, more advanced metabolites that are preferentially sequestered and retained within cells and have strong alkylating activity. Thus, the compound for use according to the present invention may be, or the composition for use according to the present invention may comprise, a metabolite that has a structure according to formula (I), (la) or (lb), wherein,

[0251] X is Ci-6 alkylene;

[0252] Wi, W2, W3 and W4 are each CH, or one of Wi, W2, W3 and W4 is N and the others are CH;

[0253] R1is selected from the group consisting of H; -Ci-4alkyl, optionally substituted by 1, 2 or 3 groups independently selected from halogen; and halogen (for example, F or Cl); R2is selected from the group consisting of H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen (for example, F or Cl); and -C1-6 alkyl optionally substituted by 1, 2 or 3 groups independently selected from halogen (for example, F or Cl); and R3is formula (Via) wherein, Rcand Rdare each independently selected from the group consisting of

[0254] H, -Ci-ealkyl, -C(O)Ci-6alkyl and -CFh-phenyl, wherein said alkyl or said phenyl is optionally substituted by 1, 2 or 3 groups selected from halogen (preferably, Rcand Rdare both H); and

[0255] Rgis selected from the group consisting of H and -Ci-ealkyl, wherein said alkyl is optionally substituted by 1, 2 or 3 groups selected from halogen (preferably Rgis H).

[0256] For the avoidance of doubt, n r in formulas (VI) and (Via) denotes the point of attachment of formula (VI) or (Via) to formula (I), (la) or (lb).

[0257] In certain embodiments, the metabolite has a structure according to formula (I), (la) or (lb), wherein,

[0258] X is Ci-6 alkylene;

[0259] R1is selected from the group consisting of H; -Ci-4alkyl; and halogen (for example, F or Cl);

[0260] R2is selected from the group consisting of H; phenyl substituted by 1, 2 or 3 halogens (for example, F or Cl); and -Ci-ealkyl substituted by 1, 2 or 3 halogens (for example, F or Cl) (for example selected from the group consisting of phenyl substituted by 1, 2 or 3 halogens (for example, F or Cl); and -Ci-ealkyl substituted by 1, 2 or 3 halogens (for example, F or Cl));

[0261] R3is formula (Via); and

[0262] Rcand Rdare both H; or a salt thereof.

[0263] In certain other embodiments, the metabolite has a structure according to formula (I), (la) or (lb), wherein,

[0264] X is Cialkylene (i.e. -CH2-) or C2alkylene (i.e. -CH2-CH2-); R1is selected from the group consisting of H; -Ci-4alkyl; and halogen (for example, F or Cl);

[0265] R2is selected from the group consisting of H; phenyl optionally substituted by 1, 2 or 3 halogens (for example, F or Cl); and -Ci-ealkyl optionally substituted by 1, 2 or 3 halogens (for example, F or Cl); and

[0266] R3is formula (Via); and

[0267] Rcand Rdare both H; or a salt thereof.

[0268] Preferably, the compound is one of formula (la) or (lb).

[0269] Thus, the compound for use according to the present invention may be, or the composition according to the invention may comprise, a compound that has a structure according to formula (I), (la) or (lb), wherein,

[0270] X is C2-ealkylene;

[0271] Wi, W2, W3 and W4 are each CH, or one of Wi, W2, W3 and W4 is N and the others are CH;

[0272] R1is selected from the group consisting of H; -Ci-4alkyl, optionally substituted by 1, 2 or 3 groups independently selected from halogen; and halogen (for example, F or Cl);

[0273] R2is selected from the group consisting of H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen (for example, F or Cl); and -Ci-ealkyl optionally substituted by 1, 2 or 3 groups independently selected from halogen (for example, F or Cl); and

[0274] R3is formula (VI) wherein, Rcand Rdare each independently selected from the group consisting of

[0275] H, -Ci-ealkyl, -C(O)Ci-6alkyl and -CH2-phenyl, wherein said alkyl or said phenyl is optionally substituted by 1, 2 or 3 groups selected from halogen (for example Rcand Rdare both H); and

[0276] Rgis selected from the group consisting of H and -Ci-ealkyl, wherein said alkyl is optionally substituted by 1, 2 or 3 groups selected from halogen (for example Rgis H).

[0277] The more advanced metabolite compounds can be used in medical treatments.

[0278] Therefore, the compound according to the invention may be, or the composition according to the invention may comprise, a compound according to formula (I), or a salt or solvate thereof wherein,

[0279] X is Ci-ealkylene;

[0280] Wi, W2, W3 and W4 are each CH, or one of Wi, W2, W3 and W4 is N and the others are CH;

[0281] R1is selected from the group consisting of H; -Ci-4alkyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and halogen;

[0282] R2is selected from the group consisting of H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and -Ci-ealkyl optionally substituted by 1, 2 or 3 groups independently selected from halogen;

[0283] R3is formula (VI) wherein, Rcand Rdare each independently selected from the group consisting of H, -Ci-ealkyl, -C(O)Ci-6alkyl and -CH2-phenyl, wherein said alkyl or said phenyl is optionally substituted by 1, 2 or 3 groups selected from halogen; and

[0284] Rgis selected from the group consisting of H and -Ci-ealkyl, wherein said alkyl is optionally substituted by 1, 2 or 3 groups selected from halogen, for use in the treatment and / or prophylaxis of venetoclax-resistant acute myeloid leukaemia.

[0285] In certain preferred embodiments, the further, more advanced metabolite is: (25)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2-yl]propanoic acid (Example compound 17);

[0286] (5)-2-amino-4-(5-(bis(2-chloroethyl)amino)-l-methyl-lH-benzo[d]imidazol-2- yl)butanoic acid (Example compound 18);

[0287] Ethyl (2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoate (Example compound 38)

[0288] (2S)-2-Amino-4-[5-[bis(2-chloroethyl)amino]-l-phenyl-benzimidazol-2-yl]butanoic acid (Example compound 41); or

[0289] (2S)-2-amino-4-[5-[bis(2-chloro-l,l,2,2-tetradeuterio-ethyl)amino]-l-methyl- benzimidazol-2-yl]butanoic acid (Example compound 49); or a salt thereof.

[0290] The metabolites described herein may be isolated from cells treated with one or more of the compounds of Formula (I) and / or may be prepared using standard organic chemistry techniques. Exemplary procedures for the preparation of metabolites of compounds of Formula (I) are described in the Examples section of International Patent Application WO2022 / 263679). For the avoidance of doubt, the metabolites described herein may be comprised in a composition of the invention in the same manner as described herein for other compounds of formula (I).

[0291] Additional

[0292] The composition of the invention may further comprise one or more additional therapeutic agents. Such further therapeutic agents may be agents useful in the treatment and / or prophylaxis of cancer, haematological cancers and, in particular, acute myeloid cancer, more particularly BCL2 inhibitor-resistant acute myeloid cancer (AML), even more particularly, venetoclax-resistant or other pharmaceutically active materials. Non-limiting examples of further therapeutic agents suitable for use in an embodiment of the present invention include: anti-apoptotic inhibitors (for example BCL-2 inhibitors, for example venetoclax itself); B-cell targeting agent such as CD20 targeting agent, for example a CD20 inhibitor, steroids (for example prednisone or dexamethasone), checkpoint inhibitors (for example CTLA-4 inhibitors or PD-1 / PD-L1 inhibitors), nuclear transport inhibitor (for example selinexor), stem cell transplants (for example autologous or allogeneic stem cell transplants), adoptive cell therapy (for example tumor-infiltrating lymphocyte (TIL) therapy, Natural Killer (NK) cell therapy, chimeric antigen receptor (CAR) T-cell therapy, and engineered T- cell receptor therapy), bispecific T-cell engagers (BiTEs), immunomodulatory imide drugs (IMiD) (for example thalidomide, lenalidomide or pomalidomide), proteasome inhibitors (Pls) (for example bortezomib, carfilzomib, ixazomib or marizomib), histone deactylase inhibitors (for example panobinostat), tubulin inhibitors (for example a taxane (for example paclitaxel or docetaxel), for example an epothilone (for example epothilone B), for example a vinca alkaloid (for example vinblastine, vinorelbine. vincristine or vindesine)), peptide drug conjugates (PDC) other than a compound of Formula (I) as described hereinabove (for example melflufen), and conventional chemotherapy, such as alkylators (for example melphalan, bendamustine, or cyclophosphamide), topoisomerase II inhibitors (for example etoposide, teniposide) and DNA intercalators (for example doxorubicin).

[0293] The one or more additional therapeutic agents may for example be selected from: a steroid, a checkpoint inhibitor, a nuclear transport inhibitor, an anti-apoptotic inhibitor, an adoptive cell therapy, a bi-specific T-cell engager, an immunomodulatory imide drug, a proteasome inhibitor, a histone deactylase inhibitor, a tubulin inhibitor, a peptide drug conjugate, an alkylator and a DNA intercalator. Preferably, the one or more additional therapeutic agents is a BCL2 inhibitor or an anti-apoptotic inhibitor. More preferably, it is venetoclax.

[0294] In an embodiment, the one or more additional therapeutic agents is a steroid, for example prednisone or dexamethasone, for example dexamethasone.

[0295] In an embodiment, the one or more additional therapeutic agents is an IMiD, for example thalidomide, lenalidomide or pomalidomide, for example lenalidomide or pomalidomide, for example pomalidomide. In an embodiment, the one or more additional therapeutic agents are an IMiD and a steroid, for example one of thalidomide, lenalidomide or pomalidomide and one of prednisone or dexamethasone, for example lenalidomide and dexamethasone or pomalidomide and dexamethasone. In an embodiment, the one or more additional therapeutic agents is a peptide drug conjugate (PDC) other than a compound of Formula (I) as described hereinabove, for example melflufen. In an embodiment, the one or more additional therapeutic agents are a peptide drug conjugate (PDC) other than a compound of Formula (I) as described hereinabove and a steroid, for example melflufen and one of prednisone or dexamethasone, for example melflufen and dexamethasone.

[0296] In an embodiment, the one or more additional therapeutic agents is an alkylator as described hereinabove, for example melphalan, bendamustine, or cyclophosphamide, for example bendamustine. In an embodiment, the one or more additional therapeutic agents are an alkylator and a steroid, for example melphalan, bendamustine, or cyclophosphamide and one of prednisone or dexamethasone, for example bendamustine and dexamethasone.

[0297] For the avoidance of doubt, the one or more further therapeutic agents may be used simultaneously, sequentially or separately with / from the administration of a composition of the invention comprising a compound of Formula (I) as described hereinabove. The composition and the one or more therapeutic agents, or the individual components of such combinations, can be administered separately at different times during the course of therapy or concurrently in divided or single combination forms. Formulations

[0298] While it is possible for a compound of Formula (I) or salts thereof, to be administered alone, it is preferable for them to be present in a formulation and particularly in a pharmaceutical formulation. Pharmaceutical formulations include those suitable for oral, parenteral (including subcutaneous, intradermal, intraosseous infusion, intramuscular, intravascular (bolus or infusion), and intramedullary), intraperitoneal, transmucosal, transdermal, rectal and topical (including dermal, buccal, sublingual and intraocular) administration although the most suitable route may depend upon, for example, the condition and disorder of the subject under treatment.

[0299] In one embodiment of the invention, a compound of Formula (I) is administered as a pharmaceutical formulation suitable for oral or parenteral (including subcutaneous, intradermal, intraosseous infusion, intramuscular, intravascular (bolus or infusion), and intramedullary) administration.

[0300] Pharmaceutical formulations of a compound of Formula (I) suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The compound of Formula (I) may also be presented as a bolus, electuary or paste. Various pharmaceutically acceptable carriers and their formulation are described in standard formulation treatises, e.g., Remington's Pharmaceutical Sciences by E. W. Martin. See also Wang, Y. J. and Hanson, M. A., Journal of Parenteral Science and Technology, Technical Report No. 10, Supp. 42:2S, 1988.

[0301] Formulations for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. Preferably the formulations may be presented in unit dosage or divided dosage containers, for example sealed ampoules and vials. The formulation may be stored in a freeze-dried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example saline, a physiologically acceptable solution or water-for-inj ection, immediately prior to use. The formulation may also be stored as a liquid pharmaceutical formulation requiring only the addition of the sterile liquid carrier, for example saline, a physiologically acceptable solution or water-for- inj ection, immediately prior to use.

[0302] Extemporaneous injection and infusion solutions and suspensions may be prepared from sterile powders, granules or other dry composition. Exemplary compositions for parenteral administration include injectable solutions or suspensions which can contain, for example, suitable non-toxic, parenterally acceptable diluents or solvents, such as mannitol, 1,3 -butanediol, water, Ringer’s solution, an isotonic sodium chloride solution, or other suitable dispersing or wetting and suspending agents, including synthetic mono- or diglycerides, and fatty acids, including oleic acid, or Cremaphor.

[0303] In a preferred embodiment of the invention, the compound of Formula (I) for use in the present invention comprises a lyophilized pharmaceutical preparation of the compound of Formula (I) or a salt thereof. The term “lyophilized pharmaceutical preparation of the compound of Formula (I) or a salt thereof' is understood to mean that the compound of Formula (I) or a salt thereof is freeze-dried ("lyophilization", "lyophilized" etc. may in the present context be used interchangeably with "freeze- drying", "freeze-dried" etc.).

[0304] In another preferred embodiment of the invention, the compound of Formula (I) for use in the present invention is in a liquid pharmaceutical preparation of the compound of Formula (I) or a salt thereof. Preferably, the liquid pharmaceutical preparation comprises or consists essentially of the following components: i) the compound of Formula (I), or a salt thereof; ii) propylene glycol; iii) optionally one or more physiologically acceptable aqueous solvent(s); and iv) optionally one or more additional therapeutic agent(s); or the liquid pharmaceutical preparation comprises or consists essentially of the following components: i) the compound of Formula (I) or melflufen, or a salt thereof; ii) polyethylene glycol; iii) optionally one or more physiologically acceptable aqueous solvent(s); and iv) optionally one or more additional therapeutic agent(s).

[0305] Preparation of liquid pharmaceutical formulations, liquid pharmaceutical formulations, and a kit for making such liquid pharmaceutical formulations of melflufen, or a salt thereof, are described in detail in WO 2020 / 212594, the contents of which are incorporated herein by reference.

[0306] It should be understood that in addition to the ingredients particularly mentioned above, the formulations for use in this invention may include other agents conventional in the art having regard to the type of formulation in question.

[0307] In a preferred embodiment of the invention is a composition consisting essentially of the compound of formula (I) as defined above and a BCL inhibitor.

[0308] In another preferred embodiment of the invention is a composition consisting essentially of the compound of formula (I) as defined above and venetoclax.

[0309] In yet another preferred embodiment of the invention is a composition consisting essentially of a metabolite, as described above of the compound of formula (I) and a BCL2 inhibitor.

[0310] In yet another preferred embodiment of the invention is a composition consisting essentially of a metabolite, as described above of the compound of formula (I) and venetoclax.

[0311] It should be understood that in addition to the ingredients particularly mentioned above, the compositions for use in this invention may include other agents conventional in the art having regard to the type of composition in question.

[0312] Kits

[0313] The invention also provides a kit comprising: a compound of formula (I) as described hereinabove.

[0314] For example, a kit of the invention may comprise a compound of formula (I) and a BCL2 inhibitor. The BCL2 inhibitor is preferably venetoclax. The invention also provides a kit comprising: a metabolite of the compound of formula (I) as described hereinabove.

[0315] For example, a kit of the invention may comprise a metabolite of the compound of formula (I) as described hereinabove and a BCL2 inhibitor. Preferably, the BCL2 inhibitor is venetoclax.

[0316] A kit of the invention may further comprise one or more additional therapeutic agents as described hereinabove. Such further therapeutic agents may be agents useful in the treatment and / or prophylaxis of acute myeloid leukaemia, more preferably BCL2 inhibitor-resistant AML, more preferably venetoclax-resistant AML or other pharmaceutically active materials. For example, a kit of the invention may further comprise one or more additional therapeutic agents which is a steroid, for example prednisone or dexamethasone, for example dexamethasone. For example, a kit of the invention may comprise one or more additional therapeutic agents which are an anti- CD20 antibody, for example rituximab, and a steroid, for example dexamethasone. For example, a kit of the invention may comprise one or more additional therapeutic agents which are an anti-CD38 antibody, for example daratumumab, and a steroid, for example dexamethasone.

[0317] A kit of the invention may further comprise one or more pharmaceutically acceptable carriers as described hereinabove.

[0318] In an embodiment of the present invention, a kit of the invention finds use in the treatment and / or prophylaxis of acute myeloid leukaemia and in particular, of venetoclax-resistant AML.

[0319] In certain embodiments of the invention, the kit of the invention comprises one or more containers and may also include sampling equipment, for example, bottles, bags (such as intravenous fluid bags), vials, syringes, and test tubes. Other components may include needles, diluents, wash reagents and buffers. Usefully, the kit may include at least one container comprising a pharmaceutically acceptable organic solvent or a pharmaceutically-acceptable buffer, such as phosphate-buffered saline, Ringer's solution and dextrose solution. Preferably, the kit of the invention comprises instructions, for example instructions that instruct a user to admix a stated amount of the compound according to formula (I) described above with a stated amount of physiologically acceptable aqueous solvent or diluent, pharmaceutically acceptable organic solvent, and / or optional one or more additional therapeutic agents. Such instructions may also provide guidance on the storage conditions and / or administration instructions.

[0320] For the avoidance of doubt, the compound according to formula (I) and the optional physiologically acceptable aqueous solvent or diluent, optional one or more pharmaceutically acceptable organic solvents, and optional one or more additional therapeutic agents, are present in a kit according to the present invention in a form and quantity suitable for the preparation of a pharmaceutical preparation according to the invention. The skilled person can readily determine a quantity of a compound according to formula (I) as described hereinabove, physiologically acceptable aqueous solvent or diluent, pharmaceutically acceptable organic solvent, and optional one or more further therapeutic agents, suitable for the use according to the present invention.

[0321] Cancers

[0322] It has been found that compounds of formula (I) as defined hereinabove display excellent in vitro cytotoxicity towards various haematological cancer cell lines. Furthermore, compounds of formula (I) described above are readily hydrolysed within cancer cells to form metabolites that are preferentially sequestered and retained within cells and have strong alkylating activity. Compounds of formula (I) are also effective at reducing tumour growth in an in ovo chicken embryo xenograft model of lymphoma.

[0323] The invention therefore provides a compound according to formula (I) as described hereinabove, for use in a method for the treatment and / or prophylaxis of venetoclax- resistant acute myeloid leukaemia (AML). For the avoidance of doubt, the compound according to formula (I), may administered with a further therapeutic agent, such as venetoclax, simultaneously, sequentially or separately from the other component and from the optional one or more additional therapeutic agents. The individual components of such combinations can be administered separately at different times during the course of therapy or concurrently in divided or single combination forms. The invention further provides a composition as described hereinabove, for example a pharmaceutical composition, or a kit as described hereinabove, for use in the treatment and / or prophylaxis of venetocl ax-resistant acute myeloid leukaemia (AML).

[0324] The invention further provides a composition as described hereinabove, for example a pharmaceutical composition, or a kit as described hereinabove, for use in the treatment and / or prophylaxis of a BCL2 inhibitor. Preferably, the BCL2 inhibitor is venetoclax-resistant acute myeloid leukemia.

[0325] In an embodiment of the invention, the compound of formula (I) the composition as described hereinabove (for example a pharmaceutical composition), or the kit as described hereinabove, find use in the treatment and / or prophylaxis of metastases of any of AML and, in particular, BCL2 inhibitor-resistant AML, more particularly venetoclax-resistant AML.

[0326] The method of the invention may further comprise administering to the subject an effective amount of one or more additional therapeutic agents as described hereinabove, for example a steroid, for example dexamethasone. In the method of the invention, the one or more additional therapeutic agents may be administered to the subject simultaneously, sequentially or separately from the compound according to formula (I). In the method of the invention, the cancer is acute myeloid leukemia and in particular, BCL2 inhibitor-resistant and more particularly venetoclax-resistant AML.

[0327] The invention further provides a method of treating or preventing a cancer as described hereinabove in a subject in need thereof, the method comprising administering to the subject an effective amount of a composition of the invention, for example a pharmaceutical composition. As such, the composition of the invention may be administered to a subject suffering, or at risk of developing acute myeloid leukaemia, particularly BCL2-inhibitor resistant AML, more particularly venetoclax- resistant AML. The method of the invention may further comprise administering to the subject an effective amount of one or more additional therapeutic agents as described hereinabove, for example a steroid, for example dexamethasone. In the method of the invention, the one or more additional therapeutic agents may be administered to the subject simultaneously, sequentially or separately from the composition of the invention.

[0328] In an embodiment of the methods of the invention, the compound according to formula (I) as described hereinabove as described hereinabove, or the composition as described hereinabove, is / are administered to a subject with a metastasis of any of the aforementioned cancers.

[0329] The invention further provides a compound of formula (I) as described hereinabove for use in the manufacture of a medicament for the treatment and / or prophylaxis of cancer, whereby the compound of formula (I) is administered to a subject suffering, or at risk of developing, acute myeloid leukaemia, particularly BCL2 inhibitor-resistant AML and more particularly venetoclax -resistant AML.

[0330] The invention further provides a compound of formula (I) as described hereinabove for use in the manufacture of a medicament for the treatment and / or prophylaxis of cancer, whereby the compound of formula (I) is administered in combination with venetoclax as described hereinabove, to a subject suffering, or at risk of developing, acute myeloid leukaemia, particularly BCL2 inhibitor-resistant AML, more particularly venetoclax-resistant AML.

[0331] Treatments

[0332] Compounds of formula (I), and metabolites derived therefrom, as described hereinabove, and compositions of the invention comprising such a compound of formula (I) find use in the treatment and / or prophylaxis of acute myeloid leukaemia, particularly BCL2 inhibitor-resistant AML, more particularly venetoclax-resistant AML.

[0333] The amounts of a compound of formula (I) as described hereinabove, or a composition of the invention comprising such a compound of formula (I), which is required to achieve a therapeutic effect will vary with the particular route of administration and the characteristics of the subject under treatment, for example the species, age, weight, sex, medical conditions, the particular disease or condition and its severity, and other relevant medical and physical factors. An ordinarily skilled physician can readily determine and administer an effective amount of a compound of formula (I) as described hereinabove required for treatment or prophylaxis of a cancer.

[0334] A compound of formula (I) as described hereinabove, or a composition of the invention comprising such a compound of formula (I), may for example be administered as a parenteral or oral dosage. Parenteral administration includes intravenous (into a vein, for example a central or a peripheral vein, bolus or infusion), intra-arterial (into an artery, for example a central or a peripheral artery), intraosseous infusion (into the bone marrow), intra-muscular (into muscle), intradermal (into the dermis), and subcutaneous (under the skin) administration. In one preferred embodiment, the dosage of the present invention is administered intravenously or intra-arterially, and, more preferably, by intravenous infusion (for example central intravenous infusion or peripheral intravenous infusion). In another preferred embodiment, the dosage of the present invention is administered by subcutaneous injection. As such, pharmaceutical formulations especially useful for the present invention are those suitable for intravenous administration, more especially intravenous infusion, or subcutaneous administration.

[0335] A compound of formula (I) as described hereinabove, or a composition of the invention comprising such a compound of formula (I), may be administered daily (including several times daily), every second or third day, weekly, every second, third, fourth, fifth or sixth week or even as a high single dose depending on the subject and disease or disorder to be treated.

[0336] A compound of formula (I) as described hereinabove, or a composition of the invention comprising such a compound of formula (I), may be administered in an amount of about 1 mg to 150 mg per administration. 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, a compound of formula (I) as described hereinabove, or a composition of the invention comprising such a compound of Formula (I), may be administered in a single high dose. A single high dose may 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.

[0337] A compound of formula (I) as described hereinabove, or a composition of the invention comprising such a compound of formula (I), may be administered with the compound of formula (I) being in an amount of about 1 mg to 150 mg per administration. 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 of the compound of formula (I). Alternatively, a compound of formula (I) as described hereinabove, or a composition of the invention comprising such a compound of formula (I), may be administered with the compound of formula (I) being in a single high dose. A single high dose may 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. In some instances, a dose is determined based on the patient’s body weight or the patient’s body surface area.

[0338] A compound of formula (I) as described hereinabove, or a composition of the invention comprising such a compound of Formula (I), may be administered as part of a treatment cycle. In a treatment cycle, the first administration is on day 1 of the cycle, wherein the cycle lasts X days, with no further administration for the next X-l days. X may be, for example, from 1 to 42, for example from 2 to 28 days, for example from 2 to 21 days, for example from 2 to 14 days. Alternatively, a compound of formula (I) as described hereinabove, or a composition of the invention comprising such a compound of formula (I), may be administered as split doses, for example on for example on days 1 and 2 of the cycle.

[0339] The cycle may be repeated one or several times depending on the category, class or stage of the cancer to be treated. For example, the cycle may be repeated from 1 to 100 times, for example from 2 to 50 times, for example 8 to 40 times, for example 8 or 16 times. For example, the compound of formula (I) as described hereinabove, or a composition of the invention comprising such a compound of Formula (I), may be administered for 8 repeats of a 7-day cycle, followed by 16 repeats of a 14 day cycle, optionally followed by further repeats of a 28 day cycle. For example, they may be administered in 21 day cycles (i.e. once every 3 weeks) until disease progression or unacceptable toxicity. An ordinarily skilled physician or clinician can readily determine the number of cycles required to prevent, counter or arrest the progress of the cancer.

[0340] The compound of formula (I) as described hereinabove and any additional therapeutic agent, as described hereinabove may be used simultaneously, sequentially or separately. For example, the individual components of such combinations can be administered separately at different times during the course of therapy or concurrently in divided or single combination forms.

[0341] Whilst a compound of Formula (I) as described hereinabove as described hereinabove may be used as the sole active ingredient in the present invention, it may also be used in combination with one or more additional therapeutic agent(s), and the use of such combinations provides one preferred embodiment of the invention. Such further therapeutic agents may be agents useful in the treatment and / or prophylaxis of cancer, or other pharmaceutically active materials. Such agents are known in the art.

[0342] Examples of further therapeutic agents suitable for use in the present invention are described hereinabove. The simultaneous, sequential or separate administration of one or additional further therapeutic agent(s) with the compound of formula (I) further enhances their effectiveness in the treatment and / or prophylaxis of cancer. An example of a preferred additional therapeutic agent is a BCL2 inhibitor. A further preferred additional therapeutic agent is venetoclax.

[0343] Preferred unit dosage compositions for use according to the invention are those containing an effective dose, or an appropriate fraction thereof, of the compound of formula I, and optionally one or more additional therapeutic agents.

[0344] The invention has been described broadly and generically herein. Those of ordinary skill in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary and that the actual parameters, dimensions, materials, and / or configurations will depend upon the specific application or applications for which the teachings of the present invention is / are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. It is, therefore, to be understood that the foregoing embodiments are presented by way of example only and that, within the scope of the appended claims and equivalents thereto, the invention may be practiced otherwise than as specifically described and claimed. The present invention is directed 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, if such features, systems, articles, materials, kits, and / or methods are not mutually inconsistent, is included within the scope of the present invention. Further, each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the invention. This includes the generic description of the invention with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein. Reference

[0345] The contents of the articles, patents, and patent applications, and all other documents and electronically available information mentioned or cited herein, 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. The applicant reserves the right physically to incorporate into this application any and all materials and information from any such articles, patents, patent applications, or other physical and electronic documents.

[0346] The following non-limiting Examples illustrate the invention.

[0347] Example Compounds: Compounds of Formula (I) as described hereinabove may be prepared according to known methods for those skilled in the art. Other reaction schemes, as well as a variety of different solvents, temperatures and other reaction conditions, could be readily devised by those skilled in the art.

[0348] Compounds of Formula (I) as described hereinabove, or metabolites thereof, may be prepared according to the protocols described in PCT application number PCT / EP2022 / 066756 which was published under the number WO2022 / 263679. The synthetic methods for Example compounds 1 to 48 as 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.

[0349] Table 1:

[0350]

[0351] 0

[0352]

[0353] Biological Examples:

[0354] Biological example 1 -

[0355] Assessment of the efficacy of Venetoclax and two peptide drug conjugates, Melflufen and the Example compound 1, in venetoclax-resistant AML patient samples by multiplexed high-throughput flow cytometry (HT-FC)

[0356] Methods 20 viably frozen acute myeloid leukemia (AML) bone marrow mononuclear cell samples, all from the relap se / refractory stage, were received from the Finnish Hematology Registry and Clinical Biobank (FHRB Biobank). The FHRB Biobank collected the samples after informed consent and using protocols approved by an ethical committee of the Helsinki University Hospital in accordance with the Declaration of Helsinki. The bone marrow mononuclear cells were isolated from the bone marrow aspirates.

[0357] High throughput flow cytometry (HT-FC) based ex vivo drug sensitivity testing was performed using iQue Screener Plus flow cytometer and ForeCyt® Software application according to the software guidelines (see www.sartorius.com). Ex vivo drug sensitivities of the cell populations were assessed using drug sensitivity scores (DSS), (see Yadav, Pemovska et al. 2014). Drug sensitivity testing of AML cell lines, both parental and with acquired resistance to venetoclax was performed using CellTiter Glow assay® 2.0 (Promega), performed according to the manufacturer’s protocol (see https: / / se.promega.com). The AML commercially available cell lines were HL-60, KASUMI-1, MOLM-13 and MV4-11. These cell lines are available to purchase from ATCC cell line bank (see https: / / www.atcc.org / products / crl-2724).

[0358] This method allowed the determination of the impact of melflufen, Example compound 1 (Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate) and venetoclax on the cell subset sensitivity levels and resistance testing on AML patient samples.

[0359] Results'. The patient cohort constituted 11 male and 9 female patients with a median age of 62 years and represented different FAB sub-types (FAB Ml(n=5); FAB M2 (n- 5); FAB M4 (n=2; FAB M5 (n=4) and missing FAB category information (n=4) (see Blood (1986) 68 (6): 1355-1362) (see Table 2 below). The patient cohort constituted distinct karyotype profiles thereby identifying the patients as having acute myeloid leukemia (AML) and, in particular venetoclax-resistant AML. The median blast cell percentage was 49% (minimum=25%; maximum=90%). The bone marrow mononuclear cell (BM-MNC) response to venetoclax, melflufen and the compound of Example 1 measured using flow cytometry (FC), revealed drug sensitivity to both peptide drug conjugates (melflufen and the Example compound 1). Venetoclax, however, exhibited a distinct drug sensitivity profile.

[0360] It was observed that the patient samples resistant to venetoclax were showing a good response to melflufen and a profound response to the Example compound 1 (see Figures 1 and 2).

[0361] Figure 1 shows a heat map showing the drug sensitivity scores (DSS) of venetoclax, melflufen and the Example compound 1 in a blast cell population for the 20 AML samples (note that the lowest 4 entries in the Venetoclax column are blue. All entries in the Compound 1 column are shades of orange to red). Figure 2 shows DSS scores for Example compound 1 and melflufen when compared to venetoclax. Here it can be seen that Example compound 1 is more sensitive than melflufen.

[0362] This observation was further validated in AML cell lines with acquired resistance to venetoclax (see Haematologica, v.108(7): 1768-1781;2023 Jul 1 (Results are shown in Figure 3) and compared to the parental cell line. Figure 3 shows the mean DSS scores for both the parental and venetoclax resistant (VenR) cell lines, HL-60, Kasumi-1, MOLM-13 and MV4-11 in response to treatment with the Example compound 1.

[0363] The same method as described above may be used to assess the efficacy of a test compound, such as Example compound 1, in combination with further potential therapeutic agent, for example a BCL2 inhibitor. A preferred BCL2 inhibitor is venetoclax.

[0364] Table 2:

[0365] Table 2 shows the characteristics of the patients as discussed above.

[0366] Biological Example 2: Identification of cellular subsets correlated to drug sensitivity and resistance using single cell RNA-sequencing (sc-RNA-seq).

[0367] Methods'. Single cell RNA-sequencing (sc-RNA-seq) was used to identify and predict cellular subsets associated with drug sensitivity and resistance. For sc-RNA-seq, libraries were prepared using the lOx Genomics Chromium® platform. The prepared libraries were then sequenced with the Illumina NovaSeq® 6000 system (see manufacturer’s instructions, https: / / emea.illumina.com / systems / sequencing-platforms / novaseq.html). lOx Genomics Cell Ranger v6.0.2 pipelines were used for data processing and analysis. The downstream analysis was performed using the Seurat tool (see manufacturer’s instructions, https: / satijalab.org / Seurat / ). Cell-type annotations were carried out using gene marker from sctype (see lanevski, A., Giri, A. K. & Aittokallio, T. Fully automated and ultra-fast cell-type identification using specific marker combinations from single-cell transcriptomica data. Nat Commun 13, 1246 (2022)) and multimodal reference mapping (see Hao, Y. et al. Integrated analysis of multimodal single-cell data. Cell 184, 3573-3587 e3529 (2021)).

[0368] Results'. A total of 17 different cell populations were identified, including different myeloid and lymphoid cell-types. In addition, the proportion of the HSC / progenitor (prog) like cells, HSC / MPP cells and neutrophils in the samples showed significant association with the drug response of the Example compound 1. Where the samples which were highly sensitive to this compound, they were found to have significantly (P-value<0.01) lower fractions of HSC / prog like cells and higher proportions of HSC / MPP cells and neutrophils (see Figures 4, 5, 6 and 7). Table 3 below shows the cell % values for these cells in the patient samples. The patient samples ranked highly-sensitive or less-sensitive to treatment with Examples compound 1.

[0369] Table 3:

[0370] Based on an ex vivo, functional assessment, the peptide drug conjugate Example compound 1, is highly active in AML, especially in models with primary or acquired resistance to venetoclax.

[0371] Figure 4 shows a single cell RNA-sequencing analysis of samples. It shows a UMAP (algorithm producing plots) projection of aggregated cells from 20 AML samples wherein each dot represents the transcriptome of a single cell and colour code defining each cluster. Figure 5 is a boxplot depicting the percentage of different celltype populations predicted in 20 AML samples where HSC / Prog-like cells were the most enriched cell-type. Figure 5 shows a complete analysis of 17 different cell-type populations that were identified, including different myeloid and lymphoid cell-types. It can be seen from Figure 5 that the HSC / Prog like cells are present in high % compared to other cell types.

[0372] Figure 6 shows Cell-types (n+3) with the significant (P< 0.01) difference in the cell % in less-sensitive and highly-sensitive sub-groups of Example compound 1.

[0373] Figure 7 shows scatter plots showing the correlation of Example compound 1 drug sensitivity scores in the blast cell population with cell percentages of (A) HSC / Prog like cell, (B) HSC / MPP cells and (C) Neutrophils. Based on the ex vivo, functional assessment, Example compound 1 is highly active in acute myeloid leukaemia primary patient samples. Furthermore, Example compound 1 is especially active in cellular models with primary or acquired resistance to venetoclax and supports the use of this drug in venetoclax -resistant AML.

Claims

Claims1. A compound according to formula (I), or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamatewherein,X is Ci-6 alkylene;Wi, W2, W3 and W4 are each CH, or one of Wi, W2, W3 and W4 is N and the others are CH;R1is selected from the group consisting of H; C1-4 alkyl, optionally substituted by 1, 2 or 3 groups independently selected from halogen; and halogen;R2is selected from the group consisting of H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and Ci-ealkyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; andR3is a group according to formula (II):or formula (III):72wherein,R4is selected from the group consisting of N(Rc)(Rd) and formula (IV):when R4is formula (IV), R5is Rb; and when R4is N(Rc)(Rd), R5is selected from the group consisting of Rband formula (V):wherein each Rais independently selected from the group consisting of H;Ci-ealkyl; -CH2-phenyl; or -CH2-3 to 12-membered heterocyclyl comprising 1, 2, 3 or 4 heteroatoms selected from N, O and S; wherein said Ci-ealkyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of -OH, -OCi-6alkyl, -NH2, -NHC(=NH)NH2, -C(O)OH, -C(O)NH2, -SH, -SCH3, and halogen; and said phenyl or heterocyclyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen, -NH2, -OH, -OCi-ealkyl and -NO2;Rbis selected from the group consisting of -OH ; -N(Re)(Rf); and -OCi-ealkyl optionally substituted by one or more groups selected from halogen, -OH, -CN, - N(Re)(Rf), -Ce-ioaryl, or 3 to 12 membered heterocycle comprising one or more O, N or S atoms and optionally substituted by 1, 2 or 3 halogens, and / or wherein said alkyl is optionally interrupted by 1, 2 or 3 O, N or S atoms;73Rcand Rdare each independently selected from the group consisting of H; -Ci-ealkyl, C(O)Ci-ealkyl; and -CLB-phenyl, wherein said alkyl or said phenyl is optionally substituted by 1, 2 or 3 groups selected from halogen; andReand Rfare each independently selected from the group consisting of H and - Ci-ealkyl, wherein said alkyl is optionally substituted by 1, 2 or 3 groups selected from halogen; or Reand Rftogether with the nitrogen atom to which they are attached form a 4-, 5- or 6-membered heterocycle which is optionally substituted by 1, 2 or 3 groups selected from halogen. for use in the treatment of and / or prophylaxis of BCL2 inhibitor resistant or refractory acute myeloid leukaemia (AML).

2. A compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate for use in the treatment of and / or prophylaxis of venetoclax -resistant amyloid myeloid leukaemia (AML).

3. A compound of formula (I) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate for use in the treatment of and / or prophylaxis of BCL2 inhibitor-resistant AML wherein there has been previous exposure to a BCL2 inhibitor.

4. The compound for use according to any one of claims 1 to 3, wherein the compound of formula (I) has the structure (la):for example, structure (lb):

745. The compound for use according to any one of the preceding claims wherein R1is H.

6. The compound for use according to any one of claims 1 to 5 wherein R2is selected from the group consisting of phenyl optionally substituted by 1, 2 or 3 halogens; and - Ci-ealkyl optionally substituted by 1, 2 or 3 halogens; for example -Ci-4alkyl optionally substituted by 1, 2 or 3 halogens, for example methyl.

7. The compound for use according to any one of claims 1 to 6 wherein X is -CH2- or -CH2-CH2-.

8. The compound for use according to any one of claims 1 to 7 wherein R3is a group according to formula (Ila):or formula (Illa):

759. The compound for use according to any one of claims 1 to 8 wherein R4is selected from the group consisting of N(Rc)(Rd) and formula (IVa):

10. The compound for use according to one of claims 1 to 9 wherein, when R4is formula (IV) or (IVa), R5is Rb; and when R4is N(Rc)(Rd), R5is selected from the group consisting of Rband formula (Va), wherein formula (Va) has the following structure,11. The compound for use according to any one of claims 1 to 10 wherein R4is NH2.

12. The compound for use according to any one of claims 1 to 11 wherein each Rais independently selected from the group consisting of Ci-ealkyl and -CFh-phenyl; wherein said Ci-ealkyl is optionally substituted by -OH; -NH2; -NHC(=NH)NH2;-C(O)OH; -C(O)NH2; -SH; -SCH3; or halogen (for example, F or Cl); and said phenyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen (for example, F or Cl); -NH2; -OH; -O-Ci-ealkyl; and - NO2.

13. The compound for use according to claim 12 wherein each Rais selected from the group consisting of -Ci-ealkyl and -CH2-phenyl; wherein said phenyl is optionally substituted by 1, 2 or 3 halogen substituents.7614. The compound for use according to any one of claims 1 to 13 wherein when R4is formula (IV) or (IVa), R5is -OCi-ealkyl; and when R4is NH2, R5is -OCi-ealkyl or formula (V) or (Va).

15. The compound for use according to claim 14 wherein when Rbis -OCi-ealkyl, it is selected from the group consisting of methoxy, ethoxy and isopropoxy.

16. The compound for use according to claim 1, wherein the compound is selected from the group consisting of:Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 1);Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-3-(4-fluorophenyl)propanoate (Example compound 2);Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-3-methyl-butanoate (Example compound 3);Ethyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]propanoyl]amino]-4-methyl-pentanoate (Example compound 4);Ethyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]propanoyl]amino]-3-(4-fluorophenyl)propanoate (Example compound 5);Ethyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]propanoyl]amino]-3-methyl-butanoate (Example compound 6);Ethyl (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-4-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]butanoate (Example compound 7);Ethyl (2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propanoyl]amino]-4-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]butanoate (Example compound 8);Ethyl (2S)-2-[[(2S)-2-amino-3-methyl-butanoyl]amino]-4-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]butanoate (Example compound 9);Ethyl (2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-3-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]propanoate (Example compound 10);Ethyl (2S)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propanoyl]amino]-3-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]propanoate (Example compound 11);Ethyl (2S)-2-[[(2S)-2-amino-3-methyl-butanoyl]amino]-3-[5-[bis(2- chloroethyl)amino]-l-methyl-benzimidazol-2-yl]propanoate (Example compound 12);77Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-phenyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 13);Methyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]propanoyl]amino]-4-methyl-pentanoate (Example compound 14); Isopropyl (2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]propanoyl]amino]-4-methyl-pentanoate (Example compound 15); ethyl (25)-2-[[(25)-2-[[(25)-2-amino-3-(4-fluorophenyl)propanoyl]amino]-4-methyl- pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoate (Example compound 16);Ethyl (2S)-2-[[(2R)-2-[[(2S)-2-amino-3-(4-fluorophenyl)propanoyl]amino]-4-methyl- pentanoyl]amino]-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoate (Example compound 21)Ethyl (2S)-2-[[(2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoyl]amino]-3-(4- fluorophenyl)propanoate (Example compound 22)Ethyl (2S)-2-[[(2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoyl]amino]-4-methyl-pentanoate (Example compound 23)2 -Morpholinoethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 24)2-Isopropoxyethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 25) Isopropyl (2 S)-2- [ [(2 S)-2-amino-4- [5 - [bi s(2-chloroethyl)amino] - 1 -methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 26) Methyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 27)3-(Dimethylamino)propyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l- methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 28)(2-Methoxy-l-methyl-ethyl) (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l- methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 29)(2S)-2-[[(2S)-2-Amino-4-[5-[bis(2-chloroethyl)amino]-l-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]-l-methyl-benzimidazol-2- yl]butanoyl]amino]-N,4-dimethyl-pentanamide (Example compound 31)Ethyl (2S)-2-[[(2S)-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2-yl]-2- (methylamino)butanoyl]amino]-4-methyl-pentanoate (Example compound 32) Ethyl (2S)-2-[[(2S)-2-acetamido-4-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]butanoyl]amino]-3-(4-fluorophenyl)propanoate (Example compound 34)Ethyl (2S)-2-[[(2S)-2-acetamido-3-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]propanoyl]amino]-4-methyl-pentanoate (Example compound 35) Ethyl (2S)-2-[[(2S)-2-acetamido-3-[5-[bis(2-chloroethyl)amino]-l-methyl- benzimidazol-2-yl]propanoyl]amino]-3-(4-fluorophenyl)propanoate (Example compound 36)Ethyl (2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoate (Example compound 38)Ethyl (2R)-2-[[(2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 45)Ethyl (2S)-2-[[(2R)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 46)Ethyl (2R)-2-[[(2R)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol- 2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 47)Ethyl (2S)-2-[[(2S)-2-amino-4-[6-[bis(2-chloroethyl)amino]-3-methyl-imidazo[4,5-b] pyridin-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 48); and Ethyl (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chloro-l,l,2,2-tetradeuterio-ethyl)amino]-l- methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoate (Example compound 50) or a pharmaceutically acceptable salt, amide or carbamate thereof, including a salt of such an amide or carbamate.

17. The compound for use according to claim 1, wherein the compound is selected from the group consisting of:(25)-2-[[(25)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoyl]amino]-3-methyl-butanoic acid (Example compound 19); and(25)-2-[[(25)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoyl]amino]-4-methyl-pentanoic acid (Example compound 20);(2S)-2-[[(2S)-4-[5-[bis(2-chloroethyl)amino]-l-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]-l- methyl-benzimidazol-2-yl]butanoic acid (Example compound 37)(2S)-2-[[(2S)-2-Amino-4-[5-[bis(2-chloroethyl)amino]-l-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]-l-phenyl-benzimidazol-2- yl]butanoyl]amino]-4-methyl-pentanoic acid (Example compound 40) 2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]propanoyl]amino]-3-methyl-butanoic acid (Example compound 42)(2S)-2-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-3-[5-[bis(2-chloroethyl)amino]-l- methyl-benzimidazol-2-yl]propanoic acid (Example compound 43)(2S)-2-[[(2S)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]propanoyl]amino]-4-methyl-pentanoic acid (Example compound 44); and (2S)-2-[[(2S)-2-amino-4-[5-[bis(2-chl oro-1, 1,2, 2-tetradeuterio-ethyl)amino]- 1- methyl-benzimidazol-2-yl]butanoyl]amino]-4-methyl-pentanoic acid (Example compound 51) or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate18. A metabolite compound formed from a compound according to any one of claims 1 to 17 for use in the treatment and / or prophylaxis of BCL2 inhibitor-resistant acute myeloid leukaemia.

19. A metabolite compound formed from a compound according to any one of claims 1 to 17 for use in the treatment and / or prophylaxis of venetoclax -resistant acute myeloid leukaemia.

20. A metabolite according to claim 18 or claim 19, wherein the metabolite is selected from the group consisting of:(25)-2-amino-3-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2-yl]propanoic acid (Example compound 17);(5)-2-amino-4-(5-(bis(2-chloroethyl)amino)-l-methyl-lH-benzo[d]imidazol-2- yl)butanoic acid (Example compound 18);Ethyl (2S)-2-amino-4-[5-[bis(2-chloroethyl)amino]-l-methyl-benzimidazol-2- yl]butanoate (Example compound 38) (2S)-2-Amino-4-[5-[bis(2-chloroethyl)amino]-l-phenyl-benzimidazol-2-yl]butanoic acid (Example compound 41); or (2S)-2-amino-4-[5-[bis(2-chloro-l,l,2,2-tetradeuterio-ethyl)amino]-l-methyl- benzimidazol-2-yl]butanoic acid (Example compound 49); or a salt thereof.

21. A composition comprising a compound for use according to any one of the preceding claims, which is preferably a pharmaceutical composition.

22. A composition comprising a compound according to any one of claims 1 to 17 for use in the treatment and / or prophylaxis of BCL2 inhibitor -resistant AML further comprising a BCL2 inhibitor or an apoptotic inhibitor.

23. A composition comprising a compound according to any one of claims 1 to 17 for use in the treatment and / or prophylaxis of venetoclax-resistant AML further comprising a BCL2 inhibitor or an apoptotic inhibitor.

24. The composition according to any one of claims 21 to 23, which further comprises one or more pharmaceutically acceptable carriers.

25. A method of treating or preventing BCL2 inhibitor-resistant acute myeloid leukaemia in a subject in need thereof, the method comprising administering to the subject a compound according to formula (I) as described in any claims 1 to 17 or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate.8126. A method of treating or preventing venetoclax-resistant acute myeloid leukaemia in a subject in need thereof, the method comprising administering to the subject a compound according to formula (I) as described in any claims 1 to 17 or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate.

27. A method of treating or preventing BCL2 inhibitor-resistant acute myeloid leukaemia in a subject in need thereof, the method comprising administering to the subject an effective amount of a composition according to any of claims 21 to 24.

28. A method of treating or preventing venetoclax-resistant acute myeloid leukaemia in a subject in need thereof, the method comprising administering to the subject an effective amount of a composition according to any of claims 21 to 24.

29. A method of treating or preventing BCL2 inhibitor-resistant acute myeloid leukaemia in a subject in need thereof, the method comprising administering to the subject an effective amount of a metabolite compound derived from the compound according to claim 1.

30. A method of treating or preventing venetoclax-resistant acute myeloid leukaemia in a subject in need thereof, the method comprising administering to the subject an effective amount of a metabolite compound derived from the compound according to claim 1.

31. A compound as described in any of claims 1 to 17 or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate for use in the manufacture of a medicament for the treatment and / or prophylaxis of BCL2 inhibitor-resistant acute myeloid leukaemia.

32. A compound as described in any of claims 1 to 17 or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamate for use in the manufacture of a medicament for the treatment and / or prophylaxis of venetoclax-resistant acute myeloid leukaemia.8233. A composition according to any of claims 21 to 24 for use in the manufacture of a medicament for the treatment and / or prophylaxis of BCL2 inhibitor-resistant acute myeloid leukaemia.

34. A composition according to any of claims 21 to 24 for use in the manufacture of a medicament for the treatment and / or prophylaxis of venetoclax-resistant acute myeloid leukaemia.

35. A metabolite compound according to any one of claims 18 to 20 for use in the manufacture of a medicament for the treatment and / or prophylaxis of BCL2 inhibitorresistant acute myeloid leukaemia.

36. A metabolite compound according to any one of claims 18 to 20 for use in the manufacture of a medicament for the treatment and / or prophylaxis of venetoclax- resistant acute myeloid leukaemia.

37. The compound or composition for use according to any one of the preceding claims, which use further comprises one or more additional therapeutic agents, for example a therapeutic agent that is selected from the group consisting of: a steroid, a checkpoint inhibitor, a nuclear transport inhibitor, a BCL2 inhibitor or an anti- apoptotic inhibitor, an adoptive cell therapy, a bi-specific T-cell engager, an immunomodulatory imide drug, a proteasome inhibitor, a histone deactylase inhibitor, a tubulin inhibitor, a peptide drug conjugate, an alkylator, a topoisomerase II inhibitor and a DNA intercalator.

38. A composition comprising according to claim 37 wherein the BCL2 inhibitor or anti-apoptotic inhibitor is venetoclax.

39. A kit comprising a compound as described in any of claims 1 to 17.

40. A kit according to claim 39, which further comprises one or more additional therapeutic agents.8341. A method of selecting a BCL2 inhibitor-resistant patient to be treated with the compound of formula (I) wherein the proportion of HSC / progenitor like cells and neutrophils determine whether a patient may be treated, wherein the patient sample tested has significantly lower fractions of HSC / progenitor like cells and higher proportions of HSC / MPP cells and neutrophils, thereby indicates sensitivity to the compound of formula (I).

42. A method of selecting a patient according to claim 41 wherein the BCL2 inhibitor is venetoclax.

43. A method of selecting a patient according to claim 41 and claim 42, wherein the HSC / Progenitor like cells are present in the range between 14 and 63% (for example between 14 and 40%), and where there is high sensitivity to the compound of formula (I).

44. A method according to claim 43 where the presence of HSC / Progenitor like cells is about 24 and where there is high sensitivity to the compound of formula (I).

45. A method of selecting a patient according to claim 31, wherein the HSC / Progenitor like cells are present in the range 15 and 63%, the HSC / MPP cells are present in the range 0 to 3.5% and the neutrophils are present in the range of 0 to 7.5% where there is low sensitivity to the compound of formula (I).

46. A method of selecting a patient according to any one of claims 41 to 45, wherein the ratio between HSC / Progenitor like cells divided by neutrophils is lower than 50%.

47. A method of selecting a patient according to claim 46 wherein the ratio between HSC / Progenitor like cells divided by neutrophils is lower than 30%.

48. A method of selecting a patient according to claim 46 wherein the ratio between HSC / Progenitor like cells divided by neutrophils is lower than 10%.

49. A method of selecting a patient according to claim 46 wherein the ratio between HSC / Progenitor like cells divided by neutrophils is lower than 3.6.8450. A method of treatment and / or prophylaxis of BCL2 inhibitor resistant or refractory acute myeloid leukaemia (AML) comprising administering an effective amount of a compound according to formula (I), or a pharmaceutically acceptable salt, ester, amide or carbamate thereof, including a salt of such an ester, amide or carbamatewherein,X is Ci-6 alkylene;Wi, W2, W3 and W4 are each CH, or one of Wi, W2, W3 and W4 is N and the others are CH;R1is selected from the group consisting of H; C1-4 alkyl, optionally substituted by 1, 2 or 3 groups independently selected from halogen; and halogen;R2is selected from the group consisting of H; phenyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; and Ci-ealkyl optionally substituted by 1, 2 or 3 groups independently selected from halogen; andR3is a group according to formula (II):or formula (III):85wherein,R4is selected from the group consisting of N(Rc)(Rd) and formula (IV):when R4is formula (IV), R5is Rb; and when R4is N(Rc)(Rd), R5is selected from the group consisting of Rband formula (V):wherein each Rais independently selected from the group consisting of H;Ci-ealkyl; -CH2-phenyl; or -CH2-3 to 12-membered heterocyclyl comprising 1, 2, 3 or 4 heteroatoms selected from N, O and S; wherein said Ci-ealkyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of -OH, -OCi-6alkyl, -NH2, -NHC(=NH)NH2, -C(O)OH, -C(O)NH2, -SH, -SCH3, and halogen; and said phenyl or heterocyclyl is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of halogen, -NH2, -OH, -OCi-ealkyl and -NO2;Rbis selected from the group consisting of -OH ; -N(Re)(Rf); and -OCi-ealkyl optionally substituted by one or more groups selected from halogen, -OH, -CN, - N(Re)(Rf), -Ce-ioaryl, or 3 to 12 membered heterocycle comprising one or more O, N or S atoms and optionally substituted by 1, 2 or 3 halogens, and / or wherein said alkyl is optionally interrupted by 1, 2 or 3 O, N or S atoms;86Rcand Rdare each independently selected from the group consisting of H; -Ci-ealkyl, C(O)Ci-ealkyl; and -Cft-phenyl, wherein said alkyl or said phenyl is optionally substituted by 1, 2 or 3 groups selected from halogen; andReand Rfare each independently selected from the group consisting of H and - Ci-ealkyl, wherein said alkyl is optionally substituted by 1, 2 or 3 groups selected from halogen; or Reand Rftogether with the nitrogen atom to which they are attached form a 4-, 5- or 6-membered heterocycle which is optionally substituted by 1, 2 or 3 groups selected from halogen.

51. A method of treatment and / or prophylaxis according to claim 50 wherein the BCL2 inhibitor resistant amyloid myeloid leukemia is venetoclax-resistant amyloid myeloid leukemia.87

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