WO2020254299A1

WO2020254299A1 - Combination of a mcl-1 inhibitor and a standard of care treatment for breast cancer, uses and pharmaceutical compositions thereof

Info

Publication number: WO2020254299A1
Application number: PCT/EP2020/066575
Authority: WO
Inventors: Sébastien BANQUET; Alix DERREAL; Florence Binlich; Dale Porter; Ensar HALILOVIC
Original assignee: Les Laboratoires Servier; Novartis Ag
Priority date: 2019-06-17
Filing date: 2020-06-16
Publication date: 2020-12-24
Also published as: AR119156A1; TW202114682A

Abstract

A combination comprising a Mcl-1 inhibitor and a second anticancer agent, wherein the second anticancer agent is selected from eribulin and fulvestrant, and compositions and uses thereof.

Description

COMBINATION OF A MCL-1 INHIBITOR AND A STANDARD OF CARE TREATMENT FOR BREAST CANCER, USES AND PHARMACEUTICAL

COMPOSITIONS THEREOF FIELD OF THE INVENTION

The present invention relates to a combination of a Mcl-1 inhibitor with a second anticancer agent, wherein the second anticancer agent is selected from eribulin (cytotoxic agent) and fulvestrant (hormonal therapy). The invention also relates to the use of said combination in the treatment of cancer, in particular breast cancer, and more particularly luminal breast cancer, HER2⁺ breast cancer and triple negative breast cancer. Also provided are pharmaceutical formulations suitable for the administration of such combinations.

BACKGROUND OF THE INVENTION

Breast cancer (BC) is the most commonly diagnosed and the second leading cause of cancer- related deaths among women worldwide (Siegel et al. CA Cancer J Clin. 2017, 67, 7-30). One of the major challenges for its treatment is its heterogeneous nature, which will determine the therapeutic options by evaluating a few biomarkers, including the presence of hormone receptors (HR) [estrogen receptor (ER) and progesterone receptor (PR)], excessive level of human epidermal growth factor receptor 2 (HER2) protein, and/or extra copies of the HER2 gene (Hammond et al. Arch. Pathol. Lab. Med. 2010, 134, e48-72; Wolff et al. J. Clin. Oncol. 2013, 31, 3997-4013).

BC is classified into four major molecular subtypes: (i) luminal A (HR⁺/HER2 ); (ii) HER2⁺; (iii) luminal B (HR⁺/HER2⁺); and (iv) triple negative (HR7HER2-). Each of these subtypes has different risk factors for incidence, therapeutic response, disease progression, and preferential organ sites of metastases.

Luminal BC positive for HR (HR⁺) represents the vast majority (60-80%) of BC cases in developed countries (American Cancer Society. Breast Cancer Facts & Figures 2017-18. Atlanta: American Cancer Society, Inc. (2017)). For HR⁺ BC, endocrine therapy is the mainstay for treatment. Sequential administration of endocrine treatments is recommended until there is a need for rapid response or evidence of clinical resistance, when chemotherapy will be indicated (Reinert et al. Ther. Adv. Med. Oncol. 2015, 7, 304-320). Since endocrine drugs work by different mechanisms, they are generally used in combination for better anticancer efficacy. However, conflicting results have been reported (Michaud et al. Oncologist 2001, 6, 538-546; Bergh et al. J. Clin. Oncol. 2012, 30, 1919-1925; Mehta et al. N. Eng. J. Med. 2012, 367, 435-444). It is generally believed that patients with endocrine therapy-naive advanced BC and those with highly endocrine- sensitive tumors may benefit the most from combination endocrine therapy (Johnston et al. Lancet Oncol. 2013, 14, 989- 998). Metastatic HR⁺ BC may develop resistance to standard hormonal therapies, which was mediated by genomic alterations in the ER and/or upregulation of other signaling pathways. Triple negative breast cancer (TNBC) is a more aggressive disease. It is associated with a poor prognosis, a high risk of local recurrence and poor disease free and overall survival. TNBC accounts for 10-15% of the BC and is associated with younger age at diagnosis, high histologic grade, and a poor short term prognosis (Lebert, Curr. Oncol. 2018, 25, S142- S150; Stevens et al. Cancer Res. 2013, 73, 2025-2030). For TNBC, standard chemotherapy remains the mainstay of treatment. Interestingly, TNBC is the BC subtype with the higher complete response rate to chemotherapy (22%). However, recurrence and metastasis rates are higher than those carrying non- TNBC tumors (Liedtke et al. J. Clin. Oncol. 2008, 26, 1275-1281). The median OS (Overall Survival) for patients with metastatic TNBC is about 9-12 months with conventional cytotoxic agents. The lack of ER, PR, and HER2 expression precludes the use of targeted therapies in advanced TNBC, and the only approved systemic treatment option is chemotherapy (usually taxanes, anthracyclines and platinum drugs (Berrada et al. Ann. Oncol. 2010, 21, vii30-35) with or without bevacizumab). However, despite an intense research in the field, new therapies for TNBC are still needed.

Apoptosis is a highly regulated cell death pathway that is initiated by various cytotoxic stimuli, including oncogenic stress and chemotherapeutic agents. It has been shown that evasion of apoptosis is a hallmark of cancer and that efficacy of many chemotherapeutic agents is dependent upon the activation of the intrinsic mitochondrial pathway (Hanahan et al. Cell 2011, 144, 646-742).

Bcl-2 (B-cell lymphoma-2) family proteins play an important role in balancing cell survival and apoptosis (Tsujimoto, Genes to Cells 1998, 3, 697-707). Three distinct subgroups of the Bcl-2 proteins control the intrinsic pathway, the pro-apoptotic BH3-only proteins (Noxa, PUMA, Bim, Bid), the pro-survival members (Bcl-2, Bcl-xL, Bcl-w, Mcl-1, Bcl-2A1) and the pro-apoptotic effector proteins (Bax and Bak) (Adams et al. Current Opinion in Immunology 2007, 19, 488-496; Czabotar et al, Nature Reviews Molecular Cell Biology 2014, 15, 49-63). Bcl-2 family proteins have been found to be dysregulated in hematological malignancies but also in solid tumors and are frequently upregulated in acquired chemoresistant cancer cells (Maji et al. Advances in Cancer Research 2017, 137, 37-75). Pro-apoptotic and anti-apoptotic signals are tightly regulated in normal breast epithelial cells. Dysregulation of this balance is required for breast tumorgenesis and to increase acquired resistance to various treatments, including targeted therapies, radiation and chemotherapies (Williams et al. Oncotarget 2014, 6, 3519-3530). In BC, differential expression of pro-survival proteins across tumor subtypes suggest that different members of this protein class could be targeted in distinct tumor subtypes (Merino et al. Sci. Transl. Med. 2017, 9, pii: eaam7049).

Bcl-2 is an estrogen-responsive gene, which is overexpressed in about 85% of ER⁺ positive BC (Dawson et al. British J Cancer 2010, 103, 668-675). Pre-clinical data using patient- derived xenograft models of ER⁺ BC suggested that intermittent dosing with a Bcl-2 inhibitor (venetoclax) synergized with tamoxifen (antiestrogenic treatment) to improve tumor response by increasing apoptosis (Vaillant et al. Cancer Cell 2013, 24, 120-129). These data recently confirmed in the clinical Phase lb study, that combining venetoclax with endocrine therapy, has a tolerable safety profile and elicits remarkable activity in ER⁺ and Bcl-2 positive BC (Lok et al. Cancer Discov. 2019, 9, 354-369).

Mcl-1 was the second member of the Bcl-2 family discovered (Kozopas et al. Proc. Natl. Acad. Sci. USA 1993, 90, 3516-3520). Mcl-1 may also be a therapeutic target because Mcl-1 amplification has been observed in a large-scale high-resolution study of somatic copy number alterations across diverse cancers, including breast cancer, and Mcl-1 can confer resistance to chemotherapy or targeted therapy (Wertz et al. Nature 2011, 471, 110-114; Placzek et al. Cell Death & Disease 2010, 1, e40). Mcl-1 appears to be the main prosurvival protein that is up-regulated in TNBC (Goodwin et al. Cell Death Differ. 2015, 22, 2098- 2106; Xiao et al. Mol. Cancer Ther. 2015, 14, 1837-1847). Moreover, Mcl-1 amplification was commonly observed in TNBC tumors that failed to achieve a complete pathological response with neoadjuvant chemotherapy (Balko et al. Cancer Discov. 2014, 4, 232-245). Finally, Mcl-1 expression was found upregulated in the antiestrogen-resistant cell lines and depletion of Mcl-1 in resistant cells caused decreased viability of cancer cells (Thrane et al. Oncogene 2015, 34, 4199-4210; Dawson et al. 2010).

Therefore, there is an obvious need to develop new agents which aim at reversing resistance to hormonal therapies. Despite advances in our understanding of the molecular basis in BC, preventing the development of resistance and identifying appropriate treatment targets for patients who have developed drug resistance are becoming increasingly important in fighting this disease. The present invention provides a novel combination of a Mcl-1 inhibitor and a second anticancer agent, wherein the second anticancer agent is eribulin and fulvestrant. The results show that the Mcl-1 inhibitor in combination with eribulin (Table 2) exhibits a strong synergistic pro-apoptotic activity in TNBC and ER⁺/HER2 breast cancer cell lines. Moreover, a Phase I/II clinical trial is currently ongoing (Example 2).

SUMMARY OF THE INVENTION

According to a first aspect of the invention, there is provided a novel combination of:

(a) a Mcl-1 inhibitor of formula (I):

wherein:

¨ D represents a cycloalkyl group, a heterocycloalkyl group, an aryl group or a heteroaryl group,

¨ E represents a furyl, thienyl or pyrrolyl ring,

¨ Xi, X₃, X₄ and X₅ independently of one another represent a carbon atom or a nitrogen atom,

¨ X2 represents a C-R26 group or a nitrogen atom,

¨ means that the ring is aromatic,

¨ Y represents a nitrogen atom or a C-R₃ group,

¨ Z represents a nitrogen atom or a C-R₄ group,

¨ Ri represents a halogen atom, a linear or branched (Ci-C₆)alkyl group, a linear or branched (C2-C6)alkenyl group, a linear or branched (C2-C6)alkynyl group, a linear or branched (Ci-C₆)haloalkyl group, a hydroxy group, a hydroxy(Ci-C₆)alkyl group, a linear or branched (Ci-C₆)alkoxy group, -S-(Ci-C₆)alkyl group, a cyano group, a nitro group, -Cys_, -(Co-C₆)alkyl-NRnRn’, -0-(Ci-C₆)alkyl-NRnRn’, -0-(Ci-C₆)alkyl-Ri₂, -C(0)-0Rn, -0-C(0)-Rn, -C(0)-NRnRn’, -NRn-C(0)-Rii’, -NRi 1 -C(0)-0Ri 1’ , -(Ci-C₆)alkyl-NRn-C(0)-Rii’, -SO2-NR11R11’, or

-S0₂-(Ci-C₆)alkyl,

¨ R2, R₃, R₄ and R₅ independently of one another represent a hydrogen atom, a halogen atom, a linear or branched (Ci-C₆)alkyl group, a linear or branched (C2-C6)alkenyl group, a linear or branched (C2-C6)alkynyl group, a linear or branched (Ci-C₆)haloalkyl, a hydroxy group, a hydroxy(Ci-C₆)alkyl group, a linear or branched (Ci-C₆)alkoxy group, a -S-(Ci-C₆)alkyl group, a cyano group, a nitro group, -(Co-C₆)alkyl-NRnRn’, -O-Cyi, -(Co-C₆)alkyl-Cyi,

-(C2-C₆)alkenyl-Cyi, -(C2-C₆)alkynyl-Cyi, -0-(Ci-C₆)alkyl-lSIRiiRii’,

-0-(Ci-C₆)alkyl-Ri2, -C(0)-0Rn, -0-C(0)-Rn, -C(0)-NRnRn’, -NRn-C(0)-Rn’, -NRi 1 -C(0)-0Ri 1’ , -(Ci-C₆)alkyl-NRn-C(0)-Rii’, -SO2-NR11R11’, or

-S0₂-(Ci-C₆)alkyl,

or the pair (Ri, R2), (R2, R3), (R3, R4), or (R4, R5) together with the carbon atoms to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that resulting ring may be substituted by from 1 to 2 groups selected from halogen, linear or branched (Ci-Ce)alkyl, -(Co-C₆)alkyl-NRnRn’, -NR13R13’, -(Co-Ce)alkyl-Cyi or oxo,

¨ ₅ and R₇ independently of one another represent a hydrogen atom, a halogen atom, a linear or branched (Ci-C₆)alkyl group, a linear or branched (C2-C6)alkenyl group, a linear or branched (C2-C6)alkynyl group, a linear or branched (Ci-C₆)haloalkyl, a hydroxy group, a linear or branched (Ci-C₆)alkoxy group, a -S-(Ci-C₆)alkyl group, a cyano group, a nitro group, -(Co-C₆)alkyl-NRnRn’, -0-(Ci-C₆)alkyl-NRiiRii\ -O-Cyi, -(Co-C₆)alkyl-Cyi, -(C2-C₆)alkenyl-Cyi,

-(C₂-C₆)alkynyl-Cyi, -0-(Ci-C₆)alkyl-Ri₂, -C(0)-ORn, -0-C(0)-Rn,

-C(0)-NRiiRn’, -NRii-C(0)-Rn’, -NRii-C(0)-ORii\

-(Ci-C₆)alkyl-NRn-C(0)-Rii’, -SO2-NR11R11’, or -S0₂-(Ci-C₆)alkyl,

or the pair (R₅, R₇), when fused with the two adjacent carbon atoms, together with the carbon atoms to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that resulting ring may be substituted by a group selected from a linear or branched (Ci-Ce)alkyl group, -NR13R13’, -(Co-Ce)alkyl-Cyi or an oxo,

¨ W represents a -CH2- group, a -NH- group or an oxygen atom,

¨ R₈ represents a hydrogen atom, a linear or branched (Ci-Cx)alkyl group, a -CHR_aR_b group, an aryl group, a heteroaryl group, an aryl(Ci-C₆)alkyl group, or a heteroaryl(Ci-C₆)alkyl group,

¨ R₉ represents a hydrogen atom, a linear or branched (Ci-Ce)alkyl group, a linear or branched (C2-C6)alkenyl group, a linear or branched (C2-C6)alkynyl group, -Cy2, -(Ci-C₆)alkyl-Cy₂, -(C2-C₆)alkenyl-Cy2, -(C2-C₆)alkynyl-Cy2, -Cy₂-Cy₃, -(C2-C₆)alkynyl-0-Cy2, -Cy2-(Co-C₆)alkyl-0-(Co-C₆)alkyl-Cy3, a halogen atom, a cyano group, -C(0)-Ri4, or -C(0)-NRi4Ri4’,

¨ Rio represents a hydrogen atom, a linear or branched (Ci-Ce)alkyl group, a linear or branched (C2-C6)alkenyl group, a linear or branched (C2-C6)alkynyl group, an aryl(Ci-C₆)alkyl group, a cycloalkyl(Ci-C₆)alkyl group, a linear or branched (Ci-Ce)haloalkyl, or -(Ci-C₆)alkyl-0-Cy4,

or the pair (R₉, Rio), when fused with the two adjacent carbon atoms, together with the carbon atoms to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen,

¨ R11 and Rn’ independently of one another represent a hydrogen atom, an optionally substituted linear or branched (Ci-Ce)alkyl group, or -(Co-Ce)alkyl-Cyi, or the pair (Rn, Rn’) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that the nitrogen in question may be substituted by from 1 to 2 groups representing a hydrogen atom, or a linear or branched (Ci-C₆)alkyl group and it being understood that one or more of the carbon atoms of the possible substituents, may be deuterated,

_¨ Ri2 represents -Cys, -Cy5-(Co-C₆)alkyl-0-(Co-C₆)alkyl-Cy₆, -Cy5-(Co-C₆)alkyl-Cy₆,

-Cy₅-(Co-C₆)alkyl-NRn-(Co-C₆)alkyl-Cy₆, -Cy₅-Cy₆-0-(Co-C₆)alkyl-Cy7,

-Cy₅-(Co-C₆)alkyl-0-(Co-C₆)alkyl-Cy9, -Cy₅-(Co-C₆)alkyl-Cy9, -NH-C(0)-NH-Ri i, -Cy₅-(Co-C₆)alkyl-NRn-(Co-C₆)alkyl-Cy₉, -C(0)-NRnRii\ -NRnRn’, -ORn, -NRii-C(0)-Rii\ -0-alkyl(Ci-Ce)-ORn, -SO2-R11, -C(0)-0Rn,

¨ Rn, R 13’ , Ri4 and Ru’ independently of one another represent a hydrogen atom, or an optionally substituted linear or branched (Ci-C₆)alkyl group,

¨ R_a represents a hydrogen atom or a linear or branched (Ci-C₆)alkyl group,

¨ R_b represents a -0-C(0)-0-Rc group, a -0-C(0)-NRcR_c’ group, or a -0-P(0)(0Rc)2 group,

¨ R_c and R_e’ independently of one another represent a hydrogen atom, a linear or branched (Ci-Cs)alkyl group, a cycloalkyl group, a (Ci-C₆)alkoxy(Ci-C₆)alkyl group, or a (Ci-C₆)alkoxycarbonyl(Ci-C₆)alkyl group,

or the pair (R_c, R_c’) together with the nitrogen atom to which they are attached form a non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from 1 to 3 heteroatoms selected from oxygen and nitrogen, it being understood that the nitrogen in question may be substituted by a group representing a linear or branched (Ci-C₆)alkyl group,

¨ Cyi, Cy2, Cy3, Cy4, Cys, Cy₆, Cy7, Cys and Cyio independently of one another, represent a cycloalkyl group, a heterocycloalkyl group, an aryl group or a heteroaryl group, ¨ Cy₉ represents or Cy₉ represents a heteroaryl group which is substituted by a group selected from -0-P(0)(OR_2O)₂; -0-P(0)(0 M⁺)₂; -(CH₂)_p-O-(CHRi8-CHRi9-O)_q-R₂₀; hydroxy; hydroxy(Ci-C₆)alkyl; -(CH₂)_r-U-(CH₂)_s-heterocycloalkyl; or -U-(CH₂)_q-NR_2iR_2i’,

¨ Ri5 represents a hydrogen atom; a -(CH₂)_p-0-(CHRi8-CHRi₉-0)_q-R₂o group; a linear or branched (Ci-C₆)alkoxy(Ci-C₆)alkyl group; a -U-(CH₂)_q-NR_2iR_2i’ group; or a - (CH₂)_r-U-(CH₂)_s-heterocycloalkyl group,

¨ Ri₆ represents a hydrogen atom; a hydroxy group; a hydroxy(Ci-C₆)alkyl group; a -(CH₂)_r-U-(CH₂)_s-heterocycloalkyl group; a (CH₂)_r-U-V-0-P(0)(OR₂o)₂ group; a -0-P(0)(0 M⁺)₂ group; a -0-S(0)₂OR₂o group; a -S(0)₂OR₂o group; a -(CH₂)_p-0-(CHRi8-CHRi₉-0)q-R₂o group; a -(CH₂)_p-0-C(0)-NR₂₂R₂₃ group; or a -U-(CH₂)_q-NR_{2 I}R₂1’ group,

¨ Ri7 represents a hydrogen atom; a -(CH₂)_p-0-(CHRi8-CHRi₉-0)_q-R₂o group; a -CH₂-0-P(0)(OR_2O)₂ group; a -0-P(0)(OR₂o)₂ group; a -0-P(0)(0 M⁺)₂ group; a hydroxy group; a hydroxy(Ci-C₆)alkyl group; a -(CH₂)_r-U-(CH₂)_s-heterocycloalkyl group; a -U-(CH₂)_q-NR_2iR_2i’ group; or an aldonic acid,

¨ M⁺ represents a pharmaceutically acceptable monovalent cation,

¨ U represents a bond or an oxygen atom,

¨ V represents a -(CH₂)_S- group or a -C(O)- group,

¨ Ri₈ represents a hydrogen atom or a (Ci-C₆)alkoxy(Ci-C₆)alkyl group,

¨ Ri9 represents a hydrogen atom or a hydroxy(Ci-C₆)alkyl group,

¨ R_2O represents a hydrogen atom or a linear or branched (Ci-C₆)alkyl group,

¨ R_2I and R_2I’ independently of one another represent a hydrogen atom, a linear or branched (Ci-C₆)alkyl group, or a hydroxy(Ci-C₆)alkyl group,

or the pair (R_2I, R_2I’) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that the resulting ring may be substituted by a group representing a hydrogen atom or a linear or branched (Ci- C₆)alkyl group,

¨ R22 represents a (Ci-C₆)alkoxy(Ci-C₆)alkyl group, a -(CH^p-NR^I group, or a -(CH₂)_P-0-(CHRi8-CHRi9-0)_q-R2o group,

¨ R23 represents a hydrogen atom or a (Ci-C₆)alkoxy(Ci-C₆)alkyl group,

or the pair (R22, R23) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 18 ring members, which may contain in addition to the nitrogen atom from 1 to 5 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that the resulting ring may be substituted by a group representing a hydrogen atom, a linear or branched (Ci- Ce)alkyl group or a heterocycloalkyl group,

¨ R24 and R24’ independently of one another represent a hydrogen atom or a linear or branched (Ci-C₆)alkyl group,

or the pair (R24, R24’) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that the resulting ring may be substituted by a group representing a hydrogen atom or a linear or branched (Ci- C₆)alkyl group,

¨ R25 represents a hydrogen atom, a hydroxy group, or a hydroxy(Ci-C₆)alkyl group,

¨ R26 represents a hydrogen atom, a halogen atom, a linear or branched (Ci-Ce)alkyl group, or a cyano group,

¨ R27 represents a hydrogen atom or a linear or branched (Ci-Ce)alkyl group,

¨ R28 represents a -0-P(0)(0 )(0 ) group, a -0-P(0)(0 )(OR3o) group, a -0-P(0)(OR_3O)(OR_3O’) group, a -(CH2)_p-0-S(0)20 group; a -(CH2)_P-S(0)20 group; a -(CH2)_p-0-S(0)20R3o group; -Cyio, a -0-C(0)-R29 group, a -0-C(0)-0R29 group or a -0-C(0)-NR2₉R2₉’ group;

¨ R29 and R29’ independently of one another represent a hydrogen atom, a linear or branched (Ci-Ce)alkyl group or a linear or branched amino(Ci-C₆)alkyl group,

¨ R30 and R3O’ independently of one another represent a hydrogen atom, a linear or branched (Ci-Ce)alkyl group or an aryl(Ci-C₆)alkyl group,

¨ R31 represents the following groups: wherein the ammonium ion optionally exists as a zwitterionic form or has a monovalent anionic counterion,

¨ n is an integer equal to 0 or 1,

¨ p is an integer equal to 0, 1, 2, or 3,

¨ q is an integer equal to 1, 2, 3 or 4,

¨ r and s are independently an integer equal to 0 or 1, it being understood that:

- "aryl" means a phenyl, naphthyl, biphenyl, indanyl or indenyl group,

"heteroaryl" means any mono- or bi-cyclic group composed of from 5 to 10 ring members, having at least one aromatic moiety and containing from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen,

"cycloalkyl" means any mono- or bi-cyclic non-aromatic carbocyclic group containing from 3 to 10 ring members,

“heterocycloalkyl” means any mono- or bi-cyclic non-aromatic carbocyclic group containing from 3 to 10 ring members, and containing from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, which may include fused, bridged or spiro ring systems, it being possible for the aryl, heteroaryl, cycloalkyl and heterocycloalkyl groups so defined and the alkyl, alkenyl, alkynyl, alkoxy, to be substituted by from 1 to 4 groups selected from optionally substituted linear or branched (Ci-C₆)alkyl, optionally substituted linear or branched (C2-C6)alkenyl group, optionally substituted linear or branched (C2-C6)alkynyl group, optionally substituted linear or branched (Ci-C₆)alkoxy, optionally substituted (Ci-C₆)alkyl-S-, hydroxy, oxo (or L -oxide where appropriate), nitro, cyano, -C(0)-OR’, -0-C(0)-R\ -C(0)-NR’R”, -NR’R”, -(C=NR’)-OR”, linear or branched

(Ci-C₆)haloalkyl, trifluoromethoxy, or halogen, it being understood that R’ and R” independently of one another represent a hydrogen atom or an optionally substituted linear or branched (Ci-C₆)alkyl group, and it being understood that one or more of the carbon atoms of the preceding possible substituents, may be deuterated, or their enantiomers, diastereoisomers, atropisomers, or addition salts thereof with a pharmaceutically acceptable acid or base, and (b) a second anticancer agent, wherein the second anticancer agent is selected from eribulin and fulvestrant,

for simultaneous, sequential or separate use.

Said compounds of formula (I), their synthesis, their use in the treatment of cancer and pharmaceutical formulations thereof, are described in WO 2015/097123, WO 2016/207216, WO 2016/207217, WO 2016/207225, WO 2016/207226, and WO 2017/125224, the contents of which are incorporated by reference.

In particular, the Mcl-1 inhibitor is Compound 1 (S64315/MIK665):

(2R)-2- { [(5L'_ί,)-5- { 3 -chloro-2-methyl-4- [2-(4-methylpiperazin- 1 -yl)ethoxy Jphenyl }-6-(4- fluorophenyl)thieno[2,3-i/]pyrimidin-4-yl]oxy }-3-(2- { [2-(2-methoxyphenyl)pyrimidin-4- yl]methoxy}phenyl)propanoic acid, or a pharmaceutically acceptable salt thereof, which is described in Example 30 of WO 2015/097123, the contents of which are incorporated by reference.

In another embodiment, the Mcl-1 inhibitor is Compound 2 (S63845):

(2R)-2- { [(5&)-5 - { 3 -chloro-2-methyl-4- [2-(4-methylpiperazin- 1 -yl)ethoxy Jphenyl }-6-(5- fluorofuran-2-yl)thieno[2,3-i/]pyrimidin-4-yl]oxy } -3-(2- { [ 1 -(2,2,2-trifluoroethyl)- 1 //- pyrazol-5-yl]methoxy}phenyl)propanoic acid, or a pharmaceutically acceptable salt thereof, which is described in Example 185 of WO 2015/097123, the contents of which are incorporated by reference.

In another embodiment, the invention provides a combination as described herein, for use in the treatment of cancer, more particularly, the treatment of breast cancer. In another embodiment, the invention provides a pharmaceutical composition comprising the combination as described herein, and at least one pharmaceutically acceptable carrier.

DEFINITIONS

‘Combination’ refers to either a fixed dose combination in one unit dosage form (e.g., capsule, tablet, or sachet), non-fixed dose combination, or a kit of parts for the combined administration where a compound of the present invention and one or more combination partners (e.g. another drug as explained below, also referred to as‘therapeutic agent’ or‘co agent’) may be administered independently at the same time or separately within time intervals, especially where these time intervals allow that the combination partners show a cooperative, e.g. synergistic effect.

The terms‘co-administration’ or‘combined administration’ or the like as utilized herein are meant to encompass administration of the selected combination partner to a single subject in need thereof (e.g. a patient), and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration or at the same time.

The term‘fixed dose combination’ means that the active ingredients, e.g. a compound of formula (I) and one or more combination partners, are both administered to a patient simultaneously in the form of a single entity or dosage.

The term‘non-fixed dose combination’ means that the active ingredients, e.g. a compound of the present invention and one or more combination partners, are both administered to a patient as separate entities either simultaneously or sequentially, with no specific time limits, wherein such administration provides therapeutically effective levels of the two compounds in the body of the patient. The latter also applies to cocktail therapy, e.g. the administration of three or more active ingredients. ‘Cancer’ means a class of disease in which a group of cells display uncontrolled growth. Cancer types include breast cancers including luminal breast cancer, HER2⁺ breast cancer and triple negative breast cancer.

‘BC’ means breast cancer.

The term‘jointly therapeutically effective’ means that the therapeutic agents may be given separately (in a chronologically staggered manner, especially a sequence-specific manner) in such time intervals that they prefer, in the warm-blooded animal, especially human, to be treated, still show a (preferably synergistic) interaction (joint therapeutic effect). Whether this is the case can, inter alia , be determined by following the blood levels, showing that both compounds are present in the blood of the human to be treated at least during certain time intervals.

‘ Standard-of-care drug’ or‘ standard-of-care chemotherapy’ means eribulin and fulvestrant.

‘Synergistically effective’ or‘synergy’ means that the therapeutic effect observed following administration of two or more agents is greater than the sum of the therapeutic effects observed following the administration of each single agent.

As used herein, the term‘treat’,‘treating’ or‘treatment’ of any disease or disorder refers in one embodiment, to ameliorating the disease or disorder (i.e., slowing or arresting or reducing the development of the disease or at least one of the clinical symptoms thereof). In another embodiment‘treat’,‘treating’ or‘treatment’ refers to alleviating or ameliorating at least one physical parameter including those which may not be discernible by the patient. In yet another embodiment,‘treat’,‘treating’ or‘treatment’ refers to modulating the disease or disorder, either physically, ( e.g ., stabilization of a discernible symptom), physiologically, ( e.g ., stabilization of a physical parameter), or both.

As used herein, a subject is‘in need of a treatment if such subject would benefit biologically, medically or in quality of life from such treatment. The term‘remission’ refers to a decrease in or disappearance of signs and symptoms of cancer.

‘Medicament’ means a pharmaceutical composition, or a combination of several pharmaceutical compositions, which contains one or more active ingredients in the presence of one or more excipients.

The term‘haloalkyl’, as used herein, refers to a linear or branched alkyl chain substituted with one or more halogen groups in place of hydrogens along the hydrocarbon chain. Examples of halogen groups suitable for substitution in the haloalkyl group include fluorine, bromine, chlorine, and iodine. Haloalkyl groups may include substitution with multiple halogen groups in place of hydrogens in an alkyl chain, wherein said halogen groups can be attached to the same carbon or to another carbon in the alkyl chain.

DETAILED DESCRIPTION OF THE INVENTION

Described below are a number of embodiments of the invention, where for convenience El is identical to the first aspect of the invention hereinabove. Further enumerated embodiments (E) of the invention are described herein. It will be recognized that features specified in each embodiment may be combined with other specified features to provide further embodiments of the present invention.

E2. A combination according to El, comprising:

(a) a Mcl-1 inhibitor of formula (I), wherein the Mcl-1 inhibitor of formula (I) is of formula (IA): wherein:

¨ Z represents a nitrogen atom or a C-R₄ group,

¨ Ri represents a linear or branched (Ci-C₆)alkyl group, a linear or branched (C2-C6)alkenyl group, a linear or branched (C2-C6)alkynyl group, a linear or branched (Ci-C₆)alkoxy group, -S-(Ci-C₆)alkyl group, a linear or branched (Ci-C₆)haloalkyl, a hydroxy group, a cyano, -NRiiRii’, -Cyx or a halogen atom,

¨ R2, R₃ and R₄ independently of one another represent a hydrogen atom, a halogen atom, a linear or branched (Ci-C₆)alkyl group, a linear or branched (C2-C6)alkenyl group, a linear or branched (C2-C6)alkynyl group, a linear or branched (Ci-C₆)haloalkyl, a hydroxy group, a linear or branched (Ci-C₆)alkoxy group, -S-(Ci-C₆)alkyl group, a cyano group, a nitro group, -(Co-C₆)alkyl-NRnRn’, -O-Cyi, -(Co-C₆)alkyl-Cyi, -(C2-C₆)alkenyl-Cyi, -(C₂-C₆)alkynyl-Cyi, -0-(Ci-C6)alkyl-NRiiRii\ -0-(Ci-C₆)alkyl-Ri₂, -C(0)-0Rn, -0-C(0)-Rn,

-C(0)-NRiiRn\ -NRii-C(0)-Rii’, -NRii-C(0)-ORii\

-(Ci-C₆)alkyl-NRii-C(0)-Rii\ -SCk-NRnRn’, or -S0₂-(Ci-C₆)alkyl,

or the pairs (R₂, R₃) or (R₃, R₄) together with the carbon atoms to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that resulting ring may be substituted by a group selected from a linear or branched (Ci-C₆)alkyl group, -NRnRn’, -(Co-C₆)alkyl-Cyi or an oxo,

¨ ₅ and R₇ independently of one another represent a hydrogen atom, a halogen atom, a linear or branched (Ci-C₆)alkyl group, a linear or branched (C2-C6)alkenyl group, a linear or branched (C2-C6)alkynyl group, a linear or branched (Ci-C₆)haloalkyl, a hydroxy group, a linear or branched (Ci-C₆)alkoxy group, a -S-(Ci-C₆)alkyl group, a cyano group, a nitro group, -(Co-C₆)alkyl-NRnRn’, -O-Cyi, -(Co-C₆)alkyl-Cyi, -(C2-C₆)alkenyl-Cyi, -(C2-C₆)alkynyl-Cyi, -0-(Ci-C₆)alkyl-Ri2, -C(0)-ORn, -0-C(0)-Rn, -C(0)-NRnRn\ -NRn-C(0)-Rii\ -NRi 1 -C(0)-ORi 1’ , -(Ci-C₆)alkyl-NRn-C(0)-Rii’, -SO2-NR11R11’, or

-S0₂-(Ci-C₆)alkyl,

or the pair (R₅, R₇), when fused with the two adjacent carbon atoms, together with the carbon atoms to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that resulting ring may be substituted by a group selected from a linear or branched (Ci-C₆)alkyl group, -NR13R13’, -(Co-C₆)alkyl-Cyi or an oxo,

¨ R₈ represents a hydrogen atom, a linear or branched (Ci-Cx)alkyl group, an aryl group, a heteroaryl group, an aryl(Ci-C₆)alkyl group, or a heteroaryl(Ci-C₆)alkyl group,

¨ R₉ represents a linear or branched (Ci-C₆)alkyl group, a linear or branched (C2-C6)alkenyl group, a linear or branched (C2-C6)alkynyl group, -Cy2, -(Ci-C₆)alkyl-Cy₂, -(C2-C₆)alkenyl-Cy2, -(C2-C₆)alkynyl-Cy2, -Cy₂-Cy₃, -(C2-C₆)alkynyl-0-Cy2, -Cy2-(Co-C₆)alkyl-0-(Co-C₆)alkyl-Cy3, a halogen atom, a cyano group, -C(0)-Ri4, or -C(0)-NRi4Ri4\

¨ R11 and Rn’ independently of one another represent a hydrogen atom, an optionally substituted linear or branched (Ci-C₆)alkyl group, or -(Co-C₆)alkyl-Cyi,

or the pair (Rn, Rn’) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that the nitrogen in question may be substituted by a group representing a hydrogen atom, or a linear or branched (Ci- Cejalkyl group and it being understood that one or more of the carbon atoms of the possible substituents, may be deuterated,

¨ R12 represents -Cys, -Cy5-(Co-C₆)alkyl-Cy₆, -Cy5-(Co-C₆)alkyl-0-(Co-C₆)alkyl-Cy₆, -Cy₅-(Co-C6)alkyl-NRii-(Co-C6)alkyl-Cy₆, -Cy₅-Cy6-0-(Co-C6)alkyl-Cy7,

-C(0)-NRiiRn’, -NRnRii’, -ORn, -NRii-C(0)-Rn\ -0-(Ci-C₆)alkyl-ORn, -SO2-R11, -C(0)-0Rn, or -NH-C(0)-NH-Rii,

¨ Ri₃, R ₁₃’ , Ri₄ and R 14’ independently of one another represent a hydrogen atom, or an optionally substituted linear or branched (Ci-Ce)alkyl group,

¨ Cyi, Cy2, Cy₃, Cys, Cy₆, Cyi and Cys independently of one another, represent a cycloalkyl group, a heterocycloalkyl group, an aryl group or a heteroaryl group,

¨ n is an integer equal to 0 or 1, it being understood that:

- "aryl" means a phenyl, naphthyl, biphenyl, indanyl or indenyl group,

“heterocycloalkyl” means any mono- or bi-cyclic non-aromatic carbocyclic group containing from 3 to 10 ring members, and containing from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, which may include fused, bridged or spiro ring systems, it being possible for the aryl, heteroaryl, cycloalkyl and heterocycloalkyl groups so defined and the alkyl, alkenyl, alkynyl, alkoxy, to be substituted by from 1 to 4 groups selected from optionally substituted linear or branched (Ci-Ce)alkyl, optionally substituted linear or branched (C2-C6)alkenyl group, optionally substituted linear or branched (C2-C6)alkynyl group, optionally substituted linear or branched (Ci-C₆)alkoxy, optionally substituted (Ci-Ce)alkyl-S-, hydroxy, oxo (or L -oxide where appropriate), nitro, cyano, -C(0)-OR’, -0-C(0)-R\ -C(0)-NR’R”, -NR’R”, -(C=NR’)-OR”, linear or branched

(Ci-Ce)haloalkyl, trifluoromethoxy, or halogen, it being understood that R’ and R” independently of one another represent a hydrogen atom or an optionally substituted linear or branched (Ci-Ce)alkyl group, and it being understood that one or more of the carbon atoms of the preceding possible substituents, may be deuterated, or their enantiomers, diastereoisomers, atropisomers, or addition salts thereof with a pharmaceutically acceptable acid or base, and (b) a second anticancer agent, wherein the second anticancer agent is selected from eribulin and fulvestrant,

for simultaneous, sequential or separate use.

E3. The combination according to El or E2, wherein the Mcl-1 inhibitor of formula (I) is of formula (IB):

wherein R9, R11, RiT and R12 are as described in El or E2 above.

E4. The combination according to El, E2 or E3, wherein the Mcl-1 inhibitor of formula (I) is Compound 1 : (2f?)-2-{[(5S^, _a)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l- yl)ethoxy]phenyl}-6-(4-fluorophenyl)thieno[2,3-£f]pyrimidin-4-yl]oxy}-3-(2-{[2-(2- methoxyphenyl)pyrimidin-4-yl]methoxy}phenyl)propanoic acid, or a pharmaceutically acceptable salt thereof.

E5. The combination according to El, E2 or E3, wherein the Mcl-1 inhibitor of formula (I) is Compound 2: (2f?)-2-{[(5S^, _a)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl) ethoxy] phenyl }-6-(5-fluorofuran-2-yl)thieno[2,3-i/]pyrimidin-4-yl]oxy }-3-(2-{ [ 1 -(2,2,2- trifluoroethyl)-l//-pyrazol-5-yl]methoxy [phenyl )propanoic acid, or a pharmaceutically acceptable salt thereof.

E6. The combination according to any of El to E5, wherein the second anticancer agent is eribulin.

E7. The combination according to any of El to E5, wherein the second anticancer agent is fulvestrant.

E8. The combination according to El, E2 or E3, comprising:

(a) Compound 1 : (2i?)-2-{[(5,S^, _a)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l- yl)ethoxy]phenyl}-6-(4-fluorophenyl)thieno[2,3-_<f]pyrimidin-4-yl]oxy}-3-(2-{[2-(2- methoxyphenyl)pyrimidin-4-yl]methoxy}phenyl)propanoic acid, or a pharmaceutically acceptable salt thereof,

and (b) a second anticancer agent, wherein the second anticancer agent is selected from eribulin and fulvestrant,

for simultaneous, sequential or separate use.

E9. The combination according to E4 or E8, wherein the dose of Compound 1 during the combination treatment is from 25 mg to 1500 mg.

E10. The combination according to E4, E8 or E9, wherein Compound 1 is administered during the combination treatment once a week. El l. A combination according to any of El to E10, wherein the Mcl-1 inhibitor is administered orally.

El 2. A combination according to any of El to E10, wherein the Mcl-1 inhibitor is administered intravenously. E13. The combination according to any of El to E10, in the form of a non- fixed dose combination.

El 4. The combination according to any of El to E10, in the form of a fixed dose combination. E15. The combination according to any of El to E14, for use in medicine.

E16. The combination according to any of El to E14, for use according to E15, wherein said use is in the treatment of cancer.

El 7. The combination according to any of El to El 4, for use according to El 6, wherein the cancer is breast cancer. El 8. The combination according to any of El to E14, for use according to any of El 5 to

El 7, wherein the Mcl-1 inhibitor and the second anticancer agent are provided in amounts which are jointly therapeutically effective for the treatment of cancer.

E19. The combination according to any of El to E14, for use according to any of E15 to El 7, wherein the Mcl-1 inhibitor and the second anticancer agent are provided in amounts which are synergistically effective for the treatment of cancer.

E20. The combination according to any of El to E14, for use according to any of El 5 to El 7, wherein the Mcl-1 inhibitor and the second anticancer agent are provided in synergistically effective amounts which enable a reduction of the dose required for each compound in the treatment of cancer, whilst providing an efficacious cancer treatment, with a reduction in side effects.

E21. A pharmaceutical composition comprising the combination according to any of El to El 4, and at least one pharmaceutically acceptable carrier.

E22. The use of a combination according to any of El to El 4, in the manufacture of a medicament for the treatment of cancer. E23. The use according to E22, wherein the cancer is breast cancer, more particularly luminal A breast cancer, luminal B breast cancer, HER2⁺ breast cancer and triple negative breast cancer.

E24. A medicament containing, separately or together,

(a) Compound 1 : (2i?)-2-{[(5,S^, _a)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl) ethoxy]phenyl}-6-(4-fluorophenyl)thieno[2,3-_<i]pyrimidin-4-yl]oxy}-3-(2-{[2-(2- methoxyphenyl)pyrimidin-4-yl]methoxy}phenyl)propanoic acid, or a pharmaceutically acceptable salt thereof,

for simultaneous, sequential or separate administration, and wherein the Mcl-1 inhibitor and the second anticancer agent are provided in effective amounts for the treatment of cancer, in particular breast cancer.

E25. A method of treating cancer, in particular breast cancer, comprising administering a jointly therapeutically effective amount of:

(a) Compound 1 : (2f?)-2-{[(5ri'_a)-5-{3-chloro-2-methyl-4-[2-(4-methylpiperazin-l-yl) ethoxy]phenyl}-6-(4-fluorophenyl)thieno[2,3-_<i]pyrimidin-4-yl]oxy}-3-(2-{[2-(2- methoxyphenyl)pyrimidin-4-yl]methoxy}phenyl)propanoic acid, or a pharmaceutically acceptable salt thereof,

and (b) a second anticancer agent, wherein the second anticancer agent is selected from eribulin and fulvestrant.

E26. A method for sensitizing a patient who is (i) refractory to at least one chemotherapy treatment, or (ii) in relapse after treatment with chemotherapy, or both (i) and (ii), wherein the method comprises administering a jointly therapeutically effective amount of (a) Mcl-1 inhibitor of formula (I) as defined in El, E2 or E3 and (b) a second anticancer agent, as described herein, to said patient.

E27. Compound 1 for use in a combination therapy with eribulin or fulvestrant, for the treatment of cancer, in particular breast cancer. E28. Eribulin for use in a combination therapy with Compound 1, for the treatment of cancer, in particular breast cancer.

E29. Fulvestrant for use in a combination therapy with Compound 1, for the treatment of cancer, in particular breast cancer.

In the pharmaceutical compositions according to the invention, the proportion of active ingredients by weight (weight of active ingredients over the total weight of the composition) is from 5 to 50 %.

Among the pharmaceutical compositions according to the invention there will be more especially used those which are suitable for administration by the oral, parenteral and especially intravenous, per- or trans-cutaneous, nasal, rectal, perlingual, ocular or respiratory route, more specifically tablets, dragees, sublingual tablets, hard gelatin capsules, glossettes, capsules, lozenges, injectable preparations, aerosols, eye or nose drops, suppositories, creams, ointments, dermal gels etc.

In one embodiment, the compound of formula (I), in particular Compound 1, is administered intravenously, for example, using the formulation as described in WO 2018/078064.

The pharmaceutical compositions according to the invention comprise one or more excipients or carriers selected from diluents, lubricants, binders, disintegration agents, stabilisers, preservatives, absorbents, colorants, sweeteners, flavourings etc.

By wav of non-limiting example there may be mentioned:

¨ as diluents lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycerol,

¨ as lubricants silica, talc, stearic acid and its magnesium and calcium salts, polyethylene glycol,

¨ as binders : magnesium aluminium silicate, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidone,

¨ as disintegrants. agar, alginic acid and its sodium salt, effervescent mixtures. The compounds of the combination may be administered simultaneously or sequentially. The administration route is preferably the intravenous infusion or injection, and the corresponding pharmaceutical compositions may allow the instantaneous or delayed release of the active ingredients. The compounds of the combination may moreover be administered in the form of two separate pharmaceutical compositions, each containing one of the active ingredients, or in the form of a single pharmaceutical composition, in which the active ingredients are in admixture.

The useful dosage regimen varies according to the sex, age and weight of the patient, the administration route, the nature of the cancer and of any associated treatments and ranges from 25 mg to 1500 mg of Mcl-1 inhibitor per week, more preferably from 50 mg to 1400 mg per week.

The dose of the second anticancer agent, as described herein, will be the same as that used when it is administered on its own. According to the current eribulin Summary of Product Characteristics (SmPC), the recommended dose of eribulin is 1.4 mg/m² which should be administered intravenously over 2 to 5 minutes on day 1 and day 8 during a 21 -day cycle. According to the current fulvestrant SmPC, the recommended dose of fulvestrant is 500 mg once monthly, with an additional 500 mg dose given two weeks after the initial dose only.

PHARMACOLOGICAL DATA

EXAMPLE 1: In vitro effect on proliferation of combining MCL-1 inhibitors with eribulin in Breast Cancer (BC) cell lines

Cell lines were sourced and maintained in the basic media supplemented with FBS as indicated in Table 1. In addition, all media contained penicillin (100 IU/mL), streptomycin (100 pg/mL) and L-glutamine (2 mM).

Cell lines were cultured at 37 °C in a humidified atmosphere containing 5% CO2 and expanded in T-150 flasks. In all cases cells were thawed from frozen stocks, expanded through > 1 passage using appropriate dilutions, counted and assessed for viability using a CASY cell counter prior to plating 150 pL/well at the densities indicated in Table 1 into 96-well plates. All cell lines were determined to be free of mycoplasma contamination in- house. Stock solutions of compounds were prepared at a concentration of 5 mM in DMSO and stored at -20 °C.

In order to analyse the activity of the compounds as single agents, cells were seeded and treated with nine 2-fold serial dilutions of each compound dispensed individually directly into the cell assay plates. Effects of the compounds on cell viability were assessed after 144 hours of incubation at 37 °C/5% CO2 by quantification of cellular ATP levels using CellTiterGlo at 75 pL reagent/well. All the experiments were performed in triplicates. Luminescence was quantified on a multipurpose plate reader. Single agent IC50S were calculated using standard four-parametric curve fitting. IC50 is defined as the compound concentration at which the CTG signal is reduced to 50% of that measured for the vehicle (DMSO) control (Table 2).

In order to analyse the activity of the compounds in combination with eribulin (Table 2), cells were seeded and treated with seven or eight 2-fold serial dilutions of each compound dispensed, either individually or in all possible permutations in a checkerboard fashion, directly into the cell assay plates. Effects of the single agents as well as their checkerboard combinations on cell viability were assessed after 144 hours of incubation at 37 °C/5% CO2 by quantification of cellular ATP levels using CellTiterGlo at 75 pL reagent/well. Two independent experiments, each one performed in duplicates, were performed. Luminescence was quantified on a multipurpose plate reader. Potential synergistic interactions between compound combinations were assessed using the Excess Inhibition 2D matrix according to the Loewe additivity model and are reported as Synergy Score (Lehar et al., Nature Biotechnology 2009, 27(7), 659-66). All calculations were performed using Chalice™ Bioinformatics Software available in Horizon website. The doubling time indicated in Table 1 is the mean of the doubling time obtained in the different passages (in T-150 flasks) performed from the thawing of the cells to their seeding in the 96-weel plates.

Table 1. Identity and assay conditions for the 8 BC cell lines used in the combination experiments.

Table 2. Synergy scores for Mcl-1 inhibitors in combination with eribulin in the indicated

BC cell lines. Interactions were deemed synergistic when scores > 3.0 where observed.

Results

The effect on cell growth of combining the Mcl-1 inhibitors of the invention with eribulin was assessed in a panel of 8 BC cell lines. Mcl-1 inhibitors as single agents inhibited the growth of the majority of the 8 BC cell lines tested (IC50 values from 50 nM to 12.4 mM - Table 2). In combination with the standard-of-care drug eribulin, synergistic growth inhibition or GI (i.e. Synergy Scores above 3 (Lehar et al, 2009)) for the majority of the cell lines tested was observed. Out of 8 cell lines, substantial cell death (i.e. GI > 150%) was measured in 5 BC cell lines upon the combined treatment of Mcl-1 inhibitors and eribulin. These data indicate that the combination of Mcl-1 inhibitors with the standard-of-care drugs for the treatment of breast cancer could provide benefit to the treatment of BC patients.

EXAMPLE 2: Clinical trial

A Phase I/II, open label, non-randomized study to evaluate safety, tolerability, pharmacokinetics and clinical activity of (2i?)-2-{[(55'_a)-5-{3-chloro-2-methyl-4-[2-(4- methylpiperazin- 1 -yl)ethoxy]phenyl} -6-(4-fluorophenyl)thieno[2,3-if]pyrimidin-4- yl]oxy}-3-(2-{[2-(2-methoxyphenyl)pyrimidin-4-yl]methoxy}phenyl)propanoic acid (Compound 1) in combination with hormonal and cytotoxic agents will be performed in patients with locally advanced or metastatic BC (EudraCT Number: 2019-000998-23 ; https://www.clinicaltrialsregister.eu/ctr-search/trial/2019-000998-23/ES).

The purpose of the Phase I multi arm study is:

as Primary Objectives, to determine the safety profile (including dose-limiting toxicities DLTs, maximum tolerated doses MTDs), tolerability and the Recommended Doses for Expansion (RDEs) of the combinations according to National Cancer Institute Common Terminology Criteria for Adverse Events (NCI- CTCAE) v5.0;

as Secondary Objectives, to determine the plasma pharmacokinetic (PK) profile and to investigate any preliminary antitumor activity of the combinations.

The purpose of the Phase II multi arm study is:

as Primary Objective, to evaluate the Objective Response Rate (ORR) of the combinations;

as Secondary Objectives, to assess the anti-tumour activity according to the Response Evaluation Criteria In Solid Tumours version 1.1 (RECIST vl.l, 2009) in terms of Response Duration (RD), Overall Survival (OS), Progression Free Survival (PFS), Best Overall Response (BOR), the safety, the tolerability and the plasma PK profiles of the combinations.

This study will be conducted in two successive parts:

1. A multi arm dose escalation Phase part: designed with three independent arms

(30 patients in each arm) that will enrol patients in parallel (three independent dose escalations will be performed). The arm of treatment will be decided by investigator taking into account type of BC the stage and previous lines of treatment.

• Arm 1: combination of Compound 1 with paclitaxel in locally advanced or metastatic TNBC or HR⁺/HER2 BC. Patient will receive Compound 1 via Intra Venous (IV) infusion, from 30 minutes and up to 3 hours depending on the patient’s body weight and toxicities observed. In the Phase I part, the dose escalation will start at 100 mg and doses up to 1000 mg might be explored. Compound 1 will be administered on D2, D9, D16 and D23 during 28-day cycle. In parallel, the same patient will receive paclitaxel at 80 mg/m² via IV infusion, as one-hour infusion, on Dl, D8 and D15 during 28-day cycle (3 weeks on and 1 week off). The dose of paclitaxel may be adapted.

• Arm 2: combination of Compound 1 with eribulin in locally advanced or metastatic TNBC or HR⁺/HER2 BC. Patient will receive Compound 1 via IV infusion, from 30 minutes and up to 3 hours depending on the patient’s body weight and toxicities observed. In the Phase I part, the dose escalation will start at 100 mg and doses up to 1000 mg might be explored. Compound 1 will be administered on D2, D9 and D16 of a 21 -day cycle. In parallel, the same patient will receive eribulin at 1.4 mg/m², via IV infusion, over 2 to 5 minutes on Dl and D8 of a 21 -day cycle in accordance with the SmPC of eribulin. The dose of eribulin may be adapted.

• Arm 3: combination of Compound 1 with fulvestrant in locally advanced or metastatic HR⁺/HER2 BC, in postmenopausal women. Patient will receive Compound 1 via IV infusion, from 30 minutes and up to 3 hours depending on the patient’s body weight and toxicities observed. In the Phase I part, the dose escalation will start at 100 mg and doses up to 1000 mg might be explored. Compound 1 will be administered on D2, D8, D15 and D22 in Cycle 1, and on Dl, D8, D15 and D22 in following cycles, during 28-day cycle. In parallel, the same patient will receive fulvestrant at 500 mg, via Intramuscular (IM) injection on Dl and D15 during 28-day cycles for the Cycle 1 and on Dl in the following cycles in accordance with the SmPC of fulvestrant. A multi arm dose expansion Phase part: designed in four parallel independent arms

at the RDE (50 patients in each arm). The arm of treatment will be decided by investigator by taking into account type of BC the stage, and previous lines of treatment.

• For the combination of Compound 1 and paclitaxel, two arms will be considered according to the disease:

^■ Arm la: combination of Compound 1 and paclitaxel in locally advanced or metastatic TNBC ^■ Arm lb: combination of Compound 1 and paclitaxel in locally advanced or metastatic HR⁺/HER2 BC

Patients will be treated at the RDE of Compound 1 + paclitaxel with the administration schedule identified during the escalation Phase I part. · Arm 2: combination of Compound 1 and eribulin in locally advanced or metastatic TNBC or HR⁺/HER2 BC. Patients will be treated at the RDE of Compound 1 + eribulin with the administration schedule identified during the escalation Phase I part.

• Arm 3: combination of Compound 1 and fulvestrant in locally advanced or metastatic HR⁺/HER2 BC, in postmenopausal women. In the expansion Phase II part, patients will be treated at the RDE of Compound 1 + fulvestrant with the administration schedule identified during the escalation Phase I part.

The total number of cycles is at the discretion of the investigator. Patients will be treated with Compound 1 in combination with paclitaxel, eribulin or fulvestrant until the disease progresses or patient’ s or physician’ s decision.

Claims

1. A combination comprising:

(a) a Mcl-1 inhibitor of formula (I):

wherein:

¨ E represents a furyl, thienyl or pyrrolyl ring,

¨ X2 represents a C-R26 group or a nitrogen atom,

¨ means that the ring is aromatic,

¨ Y represents a nitrogen atom or a C-R₃ group,

¨ Z represents a nitrogen atom or a C-R₄ group,

¨ Ri represents a halogen atom, a linear or branched (Ci-C₆)alkyl group, a linear or branched (C2-C6)alkenyl group, a linear or branched (C2-C6)alkynyl group, a linear or branched (Ci-C₆)haloalkyl group, a hydroxy group, a hydroxy(Ci-C₆)alkyl group, a linear or branched (Ci-C₆)alkoxy group, -S-(Ci-C₆)alkyl group, a cyano group, a nitro group, -Cys_, -(Co-C₆)alkyl-NRnRn’, -0-(Ci-C₆)alkyl-NRnRn’, -0-(Ci-C₆)alkyl-Ri₂, -C(0)-0Rn, -0-C(0)-Rn, -C(0)-NRnRn\ -NRn-C(0)-Rii\ -NRi 1 -C(0)-0Ri 1’ , -(Ci-C₆)alkyl-NRi i-C(0)-Ri 1’ , -SO2-NR11R11’, or -S0₂-(Ci-C₆)alkyl,

¨ Hi, R₃, R₄ and R₅ independently of one another represent a hydrogen atom, a halogen atom, a linear or branched (Ci-C₆)alkyl group, a linear or branched (C2-C6)alkenyl group, a linear or branched (C2-C6)alkynyl group, a linear or branched (Ci-C₆)haloalkyl, a hydroxy group, a hydroxy(Ci-C₆)alkyl group, a linear or branched (Ci-C₆)alkoxy group, a -S-(Ci-C₆)alkyl group, a cyano group, a nitro group, -(Co-C₆)alkyl-NRnRn’, -O-Cyi, -(Co-C₆)alkyl-Cyi,

-(C2-C₆)alkenyl-Cyi, -(C2-C₆)alkynyl-Cyi, -0-(Ci-C₆)alkyl-lSIRiiRii’,

-0-(Ci-C₆)alkyl-Ri2, -C(0)-0Rn, -0-C(0)-Rn, -C(0)-NRiiRn\ -NRn-C(0)-Rii\ -NRi 1 -C(0)-0Ri 1’ , -(Ci-C₆)alkyl-NRii-C(0)-Rii\ -SO2-NR11R11’, or

-S0₂-(Ci-C₆)alkyl,

or the pair (Ri, R2), (R2, R3), (R3, R4), or (R4, R5) together with the carbon atoms to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that resulting ring may be substituted by from 1 to 2 groups selected from halogen, linear or branched

(Ci-Ce)alkyl, -(Co-Cejalkyl-NRnRn’, -NR13R13’, -(Co-Ce)alkyl-Cyi or oxo,

¨ ₅ and R₇ independently of one another represent a hydrogen atom, a halogen atom, a linear or branched (Ci-C₆)alkyl group, a linear or branched (C2-C6)alkenyl group, a linear or branched (C2-C6)alkynyl group, a linear or branched

(Ci-C₆)haloalkyl, a hydroxy group, a linear or branched (Ci-C₆)alkoxy group, a -S-(Ci-C₆)alkyl group, a cyano group, a nitro group, -(Co-C₆)alkyl-NRnRn’, -0-(Ci-C₆)alkyl-NRiiRii\ -O-Cyi, -(Co-C₆)alkyl-Cyi, -(C2-C₆)alkenyl-Cyi, -(C2-C₆)alkynyl-Cyi, -0-(Ci-C₆)alkyl-Ri₂, -C(0)-0Rn, -0-C(0)-Rn,

-C(0)-NRiiRn’, -NRii-C(0)-Rn’, -NRii-C(0)-ORii\

-(Ci-C₆)alkyl-NRii-C(0)-Rii’, -SO2-NR11R11’, or -S0₂-(Ci-C₆)alkyl,

or the pair (R₅, R₇), when fused with the two adjacent carbon atoms, together with the carbon atoms to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that resulting ring may be substituted by a group selected from a linear or branched (Ci-C₆)alkyl group, -NR₁₃R₁₃’, -(Co-Ce)alkyl-Cyi or an oxo, ¨ W represents a -CH2- group, a -NH- group or an oxygen atom,

¨ R-₈ represents a hydrogen atom, a linear or branched (Ci-Cx)alkyl group, a -CHR_aR_b group, an aryl group, a heteroaryl group, an aryl(Ci-C₆)alkyl group, or a heteroaryl(Ci-C₆)alkyl group,

¨ R₉ represents a hydrogen atom, a linear or branched (Ci-C₆)alkyl group, a linear or branched (C2-C6)alkenyl group, a linear or branched (C2-C6)alkynyl group, -Cy2, -(Ci-C₆)alkyl-Cy₂, -(C2-C₆)alkenyl-Cy2, -(C2-C₆)alkynyl-Cy2, -Cy₂-Cy₃, -(C2-C₆)alkynyl-0-Cy2, -Cy2-(Co-C₆)alkyl-0-(Co-C₆)alkyl-Cy3, a halogen atom, a cyano group, -C(0)-Ri4, or -C(0)-NRi4Ri4’,

¨ R11 and Rn’ independently of one another represent a hydrogen atom, an optionally substituted linear or branched (Ci-Ce)alkyl group, or -(Co-Ce)alkyl-Cyi,

or the pair (Rn, Rn’) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that the nitrogen in question may be substituted by from 1 to 2 groups representing a hydrogen atom, or a linear or branched (Ci-Ce)alkyl group and it being understood that one or more of the carbon atoms of the possible substituents, may be deuterated,

_¨ R12 represents -Cys, -Cy5-(Co-C₆)alkyl-0-(Co-C₆)alkyl-Cy₆, -Cy5-(Co-C₆)alkyl-Cy₆,

-Cy₅-(Co-C₆)alkyl-NRn-(Co-C₆)alkyl-Cy₆, -Cys-Cye-CMCo-Cejalkyl-Cyv,

-Cy₅-(Co-C₆)alkyl-0-(Co-C₆)alkyl-Cy₉, -Cy₅-(Co-C₆)alkyl-Cy₉, -NH-C(0)-NH-Ri 1, -Cy₅-(Co-C₆)alkyl-NRn-(Co-C₆)alkyl-Cy₉, -C(0)-NRnRii\ -NRiiRn’, -ORn, -NRii-C(0)-Rii\ -0-alkyl(Ci-Ce)-ORn, -SO2-R11, -C(0)-ORn,

¨ Rn, R₁₃’, Ri₄ and Ru’ independently of one another represent a hydrogen atom, or an optionally substituted linear or branched (Ci-C₆)alkyl group,

¨ R_a represents a hydrogen atom or a linear or branched (Ci-C₆)alkyl group,

¨ R_c and R_e’ independently of one another represent a hydrogen atom, a linear or branched (C i-Cx)alkyl group, a cycloalkyl group, a (Ci-C₆)alkoxy(Ci-C₆)alkyl group, or a (Ci-C₆)alkoxycarbonyl(Ci-C₆)alkyl group,

¨ Cyi, Cy2, Cy3, Cy4, Cys, Cy6, Cy7, Cys and Cyio independently of one another, represent a cycloalkyl group, a heterocycloalkyl group, an aryl group or a heteroaryl group,

¨ Cy₉ represents

or Cy₉ represents a heteroaryl group which is substituted by a group selected from -0-P(0)(OR_2O)₂; -0-P(0)(0 M⁺)₂; -(CH₂)p-0-(CHRi₈-CHRi9-0)_q-R2o; hydroxy; hydroxy(Ci-C₆)alkyl; -(CH2)_r-U-(CH2)_s-heterocycloalkyl; or -U-(CH2)_q-NR2iR2i\

¨ Ri5 represents a hydrogen atom; a -(CH2)_p-0-(CHRi8-CHRi9-0)_q-R2o group; a linear or branched (Ci-C₆)alkoxy(Ci-C₆)alkyl group; a -U-(CH2)_q-NR2iR2i’ group; or a - (CH2)_r-U-(CH2)_s-heterocycloalkyl group,

¨ Ri₆ represents a hydrogen atom; a hydroxy group; a hydroxy(Ci-C₆)alkyl group; a -(CH2)r-U-(CH2)_s-heterocycloalkyl group; a (CH2)_r-U-V-0-P(0)(OR2o)2 group; a -0-P(0)(0 M⁺)2 group; a -0-S(0)20R2o group; a -S(0)20R2o group; a -(CH₂)_P-0-(CHRi8-CHRi9-0)_q-R2o group; a -(CH2)_p-0-C(0)-NR22R23 group; or a -U-(CH₂)_q-NR21R21’ group,

¨ Ri7 represents a hydrogen atom; a -(CH2)_p-0-(CHRi8-CHRi9-0)_q-R2o group; a -CH₂-0-P(0)(OR_2O)2 group; a -0-P(0)(OR₂o)₂ group; a -0-P(0)(0 M⁺)₂ group; a hydroxy group; a hydroxy(Ci-C₆)alkyl group; a -(CH₂)_r-U-(CH₂)_s-heterocycloalkyl group; a -U-(CH₂)q-NR_2lR_2l ^, group; or an aldonic acid,

¨ M⁺ represents a pharmaceutically acceptable monovalent cation,

¨ U represents a bond or an oxygen atom,

¨ V represents a -(CH₂)_S- group or a -C(O)- group,

¨ Ri₈ represents a hydrogen atom or a (Ci-C₆)alkoxy(Ci-C₆)alkyl group,

¨ Ri9 represents a hydrogen atom or a hydroxy(Ci-C₆)alkyl group,

¨ R₂O represents a hydrogen atom or a linear or branched (Ci-C₆)alkyl group,

¨ R₂₂ represents a (Ci-C₆)alkoxy(Ci-C₆)alkyl group, a -(CH₂)p-NR₂4R₂4’ group, or a -(CH₂)p-0-(CHRi8-CHRi₉-0)q-R₂o group,

¨ R₂₃ represents a hydrogen atom or a (Ci-C₆)alkoxy(Ci-C₆)alkyl group,

or the pair (R₂₂, R^) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 18 ring members, which may contain in addition to the nitrogen atom from 1 to 5 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that the resulting ring may be substituted by a group representing a hydrogen atom, a linear or branched (Ci- Ce)alkyl group or a heterocycloalkyl group,

¨ R₂₄ and R₂₂’ independently of one another represent a hydrogen atom or a linear or branched (Ci-C₆)alkyl group,

or the pair (R₂4, R_M’) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring composed of from 5 to 7 ring members, which may contain in addition to the nitrogen atom from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, it being understood that the resulting ring may be substituted by a group representing a hydrogen atom or a linear or branched (Ci- C₆)alkyl group,

¨ His represents a hydrogen atom, a hydroxy group, or a hydroxy(Ci-C₆)alkyl group,

¨ Hie represents a hydrogen atom, a halogen atom, a linear or branched (Ci-C₆)alkyl group, or a cyano group,

¨ Hi_? represents a hydrogen atom or a linear or branched (Ci-C₆)alkyl group,

¨ His represents a -0-P(0)(0 )(0 ) group, a -0-P(0)(0 )(OR₃o) group, a -0-P(0)(OR3O)(OR3O’) group, a -(CH2)_p-0-S(0)20 group; a -(CH2)_P-S(0)20 group; a -(CH2)_p-0-S(0)20R₃o group; -Cyio, a -0-C(0)-R2₉ group, a -0-C(0)-0R2₉ group or a -0-C(0)-NR2₉R2₉’ group;

¨ R29 and R29’ independently of one another represent a hydrogen atom, a linear or branched (Ci-C₆)alkyl group or a linear or branched amino(Ci-C₆)alkyl group,

¨ R30 and R3_O’ independently of one another represent a hydrogen atom, a linear or branched (Ci-C₆)alkyl group or an aryl(Ci-C₆)alkyl group,

¨ R31 represents the following groups:

wherein the ammonium ion optionally exist as a zwitterionic form or has a monovalent anionic counterion,

¨ n is an integer equal to 0 or 1,

¨ p is an integer equal to 0, 1, 2, or 3,

¨ q is an integer equal to 1, 2, 3 or 4,

- "aryl" means a phenyl, naphthyl, biphenyl, indanyl or indenyl group,

"heteroaryl" means any mono- or bi-cyclic group composed of from 5 to 10 ring members, having at least one aromatic moiety and containing from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, "cycloalkyl" means any mono- or bi-cyclic non-aromatic carbocyclic group containing from 3 to 10 ring members,

“heterocycloalkyl” means any mono- or bi-cyclic non-aromatic carbocyclic group containing from 3 to 10 ring members, and containing from 1 to 3 heteroatoms selected from oxygen, sulphur and nitrogen, which may include fused, bridged or spiro ring systems, it being possible for the aryl, heteroaryl, cycloalkyl and heterocycloalkyl groups so defined and the alkyl, alkenyl, alkynyl, alkoxy, to be substituted by from 1 to 4 groups selected from optionally substituted linear or branched (Ci-C₆)alkyl, optionally substituted linear or branched (C2-C6)alkenyl group, optionally substituted linear or branched (C2-C6)alkynyl group, optionally substituted linear or branched (Ci-C₆)alkoxy, optionally substituted (Ci-C₆)alkyl-S-, hydroxy, oxo (or L -oxide where appropriate), nitro, cyano, -C(0)-OR’, -0-C(0)-R’, -C(0)-NR’R”, -NR’R”, -(C=NR’)-OR”, linear or branched (Ci-C₆)haloalkyl, trifluoromethoxy, or halogen, it being understood that R’ and R’’ independently of one another represent a hydrogen atom or an optionally substituted linear or branched (Ci-Ce)alkyl group, and it being understood that one or more of the carbon atoms of the preceding possible substituents, may be deuterated, or their enantiomers, diastereoisomers, atropi somers, or addition salts thereof with a pharmaceutically acceptable acid or base, and (b) a second anticancer agent, wherein the second anticancer agent is selected from eribulin and fulvestrant,

for simultaneous, sequential or separate use.

2. The combination according to claim 1, wherein the Mcl-1 inhibitor of formula (I) is of formula (IB): wherein R9, R11, RiT and R12 are as described in claim 1 above.

3. The combination according to claim 1 or claim 2, wherein the Mcl-1 inhibitor of formula (I) is Compound 1 : (2f?)-2-{[(5&)-5-{3-chloro-2-methyl-4-[2-(4- methylpiperazin-l-yl)ethoxy]phenyl}-6-(4-fluorophenyl)thieno[2,3-i/]pyrimidin-4- yl]oxy}-3-(2-{[2-(2-methoxyphenyl)pyrimidin-4-yl]methoxy}phenyl)propanoic acid, or a pharmaceutically acceptable salt thereof.

4. The combination according to claim 1 or claim 2, wherein the Mcl-1 inhibitor of formula (I) is Compound 2: (2f?)-2-{[(5&)-5-{3-chloro-2-methyl-4-[2-(4- methylpiperazin-l-yl)ethoxy]phenyl}-6-(5-fluorofuran-2-yl)thieno[2,3-i/]pyrimidin-4- yl]oxy }-3-(2-i [ l-(2,2,2-trifluoroethyl)- l//-pyrazol-5-yl]methoxy }phenyl)propanoic acid, or a pharmaceutically acceptable salt thereof.

5. The combination according to claim 1, wherein the second anticancer agent is eribulin.

6. The combination according to claim 1, wherein the second anticancer agent is fulvestrant.

7. The combination according to any one of claims 1 to 6, in the form of a non-fixed dose combination.

8. The combination according to any one of claims 1 to 7, for use in the treatment of cancer.

9. The combination according to any one of claims 1 to 7, for use according to claim 8, wherein the cancer is breast cancer.

10. The combination according to any one of claims 1 to 7, for use according to claim 8 or claim 9, wherein the Mcl-1 inhibitor and the second anticancer agent are provided in amounts which are synergistically effective for the treatment of cancer.

11. A pharmaceutical composition comprising the combination according to any one of claims 1 to 7, and at least one pharmaceutically acceptable carrier.

12. The use of a combination according to any one of claims 1 to 7, in the manufacture of a medicament for the treatment of cancer.