WO2014180882A2

WO2014180882A2 - Treatment of brain metastasis from cancer

Info

Publication number: WO2014180882A2
Application number: PCT/EP2014/059303
Authority: WO
Inventors: Angels SIERRA JIMÉNEZ; Baldomero Oliva Miguel; Laia MUIXÍ
Original assignee: Universitat de Barcelona UB
Current assignee: Universitat de Barcelona UB
Priority date: 2013-05-07
Filing date: 2014-05-07
Publication date: 2014-11-13
Anticipated expiration: 2015-11-07
Also published as: WO2014180882A3

Abstract

The present invention discloses the use of compounds of formula (I), GRP94 inhibitors and HDAC inhibitors in the treatment of brain metastasis from cancer, preferably brain metastasis from breast cancer or brain metastasis from lung cancer.

Description

TREATMENT OF BRAIN METASTASIS FROM CANCER

Field of the Invention

The present invention relates to methods and compositions for the treatment of brain metastasis from cancer, preferably brain metastasis from breast cancer or from lung cancer.

Background of the Invention

A brain metastasis is a secondary tumor induced by cancer cells that has migrated to the brain from another location in the body. Many tumor or cancer types can spread to the brain, being lung cancer and breast cancer among the most common sources of brain metastases. Metastatic brain tumors occur in about 25% of all cancers that spread through the body and are much more common than primary brain tumors.

The treatment of brain metastasis remains challenging. Whole brain radiation therapy (WBRT) is the standard treatment for patients with multiple brain metastases or with a life expectancy of less than three months. Side effects of WBRT can include memory loss, extreme fatigue, confusion, lack of coordination or even dementia. Surgical resection of the tumor prolongs survival only in patients having a single lesion in an area of the brain where it is safe to operate. Chemotherapy has not been extensively studied for brain metastasis as is generally less efficient than surgery or radiation. Corticosteroids or steroids are administered to reduce edema, however side effects from steroids can be very serious.

Effective and well tolerated therapy for brain metastasis remains an unmet medical need.

Summary of the Invention

The inventors have identified new compounds useful in the prevention and treatment of brain metastasis from cancer.

Therefore, in a first aspect, the invention is directed to a compound of formula (I)

(I) wherein

X is selected from -CH₂- and -C(O)-;

R¹ is independently selected from halogen, Ci-C₆ alkyl, CrC₆haloalkyl and NR'R", wherein R' and R" are independently selected from H, and CrC₆ alkyl;

R² is selected from H and Ci-C₆ alkyl; and

n is an integer selected from 0, 1 , 2, 3 and 4;

or a pharmaceutically acceptable salt, isomer or solvate thereof,

for use in the prevention or treatment of brain metastasis from cancer.

In another aspect, the invention is directed to a GRP94 inhibitor compound for use in the prevention or treatment of brain metastasis from cancer.

Another aspect refers to a HDAC inhibitor compound for use in the prevention or treatment of brain metastasis from cancer.

In another aspect, the invention refers to a combination comprising a compound of formula (I), or a pharmaceutically acceptable salt, isomer or solvate thereof, and a GRP94 inhibitor, for simultaneous, separate or sequential use in the prevention or treatment of brain metastasis from cancer.

In a further aspect, the invention refers to a pharmaceutical composition comprising a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, isomer or solvate thereof, a GRP94 inhibitor and a pharmaceutically acceptable carrier.

In another aspect, the invention is directed to the use of a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, isomer or solvate thereof, in the preparation of a medicament for the prevention or treatment of brain metastasis from cancer.

In another aspect, the invention refers to the use of a GRP94 inhibitor compound in the preparation of a medicament for the prevention or treatment of brain metastasis from cancer.

In another aspect, the invention refers to the use of a HDAC inhibitor compound in the preparation of a medicament for the prevention or treatment of brain metastasis from cancer.

In another aspect, the invention is directed to the use of a compound of formula (I), or a pharmaceutically acceptable salt, isomer or solvate thereof, in the preparation of a medicament for the prevention or treatment of brain metastasis from cancer by simultaneous, separate or sequential combination therapy with a GRP94 inhibitor.

In another aspect, the invention is directed to the use of a GRP94 inhibitor compound in the preparation of a medicament for the prevention or treatment of brain metastasis from cancer by simultaneous, separate or sequential combination therapy with a compound of formula (I), or a pharmaceutically acceptable salt, isomer or solvate thereof.

In another aspect the invention refers to a method of preventing or treating brain metastasis from cancer in a patient, said method comprising the administration of an effective amount of a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, isomer or solvate thereof.

Another aspect refers to a method of preventing or treating brain metastasis from cancer in a patient, said method comprising the administration of an effective amount of a GRP94 inhibitor compound.

In another aspect the invention is directed to a method of preventing or treating brain metastasis from cancer in a patient, said method comprising the administration of an effective amount of a HDAC inhibitor compound.

In a further aspect the invention is directed to a method of preventing or treating brain metastasis from cancer in a patient, said method comprising the simultaneous, separate or sequential administration of an effective amount of a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, isomer or solvate thereof, and a GRP94 inhibitor compound.

Brief Description of the Figures

Figure 1 is a graph showing the percentage of cell survival 72 h after 17-AAG treatment in the breast cancer metastatic variants in brain MDA-MB 435-P (P), 435Br1 , V5, V5CA1 and 361 . Survival is expressed as a percentage of cells with regard to control cells without treatment.

Figure 2 shows A) Western-blot showing GRP94 expression of 435/P, 435/Br1 and two more brain metastatic cells, Br1V5 and Br1 V5CA1 , derived from in vivo/in vitro successive rounds to obtain high brain metastatic cells; and B) Survival assay using Br1V5 to assess the IC50 of the NVP, which exerted a cytotoxic effect with a IC50 at 50 nM. Figure 3 is a time-course diagram showing the mean growth ratio of the tumors (volume/mm³) after administration of vehicle (control), 17-AAG (AAG17), NVP-AUY922 (NVP), docetaxel (TXT - comparative) and lenalidomide (LND).

Figure 4. is a time-course diagram showing the mean growth ratio of the tumors (volume/mm³) after administration of vehicle (control), 17-AAG (AAG17), docetaxel (TXT - comparative) and sodium valproate (VAL).

Figure 5 is a time-course diagram showing the mean growth ratio of the tumors (volume/mm³) after administration of vehicle (control), NVP-AUY922 (NVP), docetaxel (TXT - comparative), lenalidomide (LND), the combination of lenalidomide and docetaxel (LND+TXT) and the combination of lenalidomide and NVP-AUY922 (LND+NVP).

Detailed Description of the Invention

Compounds of formula (I)

In an aspect, the invention is directed to a compound (I) as defined above, or a pharmaceutically acceptable salt, isomer or solvate thereof, for use in the prevention or treatment of brain metastasis from cancer, preferably brain metastasis from lung cancer or brain metastasis from breast cancer.

In the context of the invention, the term "CrC₆ alkyl" refers to a saturated branched or linear alkyl chain which contains from 1 to 6, preferably from 1 to 3 (CrC₃ alkyl), carbon atoms and which is bound to the rest of the molecule through a single bond. Illustrative examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, etc. Preferably, Ci-C₆ alkyl and C1-C₃ alkyl are selected from methyl, ethyl and propyl groups.

Halogen refers to fluor, chloro, bromo and iodo.

Haloalkyl refers to a Ci-C₆ alkyl, preferably a C1-C₃ alkyl, as defined above wherein at least one hydrogen atom has been replaced by halogen. Illustrative examples include CF₃, CH₂F, CHF₂, CF₂CF₃, CH₂Br, CHBr₂, CCI₃, CHCI₂, CH₂CI, and the like. Preferably haloalkyl refers to CF₃.

In a particular embodiment, each R¹ is independently selected from halogen, d-

C₃ alkyl, Ci-C₃haloalkyl and NR'R", wherein R' and R" are independently selected from H, and C C₃ alkyl. In a preferred embodiment, R¹ is NR'R", wherein R' and R" are independently selected from H, and Ci-C₆ alkyl. More preferably, R' and R" are independently selected from H, and C1-C3 alkyl.

In a more preferred embodiment, R¹ is NH₂.

In a particular embodiment, n is selected from 0, 1 and 2. Preferably, n is selected from 0 and 1.

In another embodiment, R¹ is NH₂ and n is selected from 0 and 1 .

In a particular embodiment, R² is selected from H and Ci-C₃ alkyl. Preferably, R² is H.

In a preferred embodiment, R¹ is NH₂, R² is H, and n is selected from 0 and 1.

In a more preferred embodiment, the compound of formula (I) is selected from thalidomide, lenalidomide and pomalidomide, or a pharmaceutically acceptable salt, isomer or solvate thereof. In more preferred embodiment, the compound of formula (I) is lenalidomide or a pharmaceutically acceptable salt, isomer or solvate thereof. In another preferred embodiment, the compound of formula (I) is pomalidomide or a pharmaceutically acceptable salt, isomer or solvate thereof.

Preferably, the compounds of formula (I) are for use in the treatment or prevention of brain metastasis from cancer, preferably brain metastasis from lung cancer or breast cancer, in patients overexpressing the Fn14 protein.

GRP94 inhibitor compounds

In another aspect, the invention is directed to a GRP94 inhibitor compound for use in the prevention or treatment of brain metastasis from cancer, preferably brain metastasis from lung cancer or brain metastasis from breast cancer.

The term "GRP94 inhibitor" refers to compounds that exhibit inhibition of Glucose-regulated protein 94 (GRP94). Non-limiting examples of GRP94 inhibitors include, for example, 17-(allylamino)-17-demethoxygeldanamycin (17-AAG), 5-(2,4- dihydroxy-5-isopropyl-phenyl)-N-ethyl-4-[4-(morpholinomethyl)phenyl]isoxazole-3- carboxamide (NVP-AUY922), 17-dimethylaminoethylamino-17- demethoxygeldanamycin (17-DMAG), geldanamycin, radicicol, herbimycin A, celastrol, 5'-N-ethylcarboxamideadenosine (NECA) and novoiocin, or a pharmaceutically acceptable salt, isomer or solvate thereof. These compounds are also known as HSP90 (heat shock protein 90) inhibitors. In a particular embodiment, the GRP94 inhibitor is selected from 17-AAG and NVP-AUY922, or a pharmaceutically acceptable salt, isomer or solvate thereof. In an embodiment, the GRP94 inhibitor is 17-AAG or a pharmaceutically acceptable salt, isomer or solvate thereof. In another embodiment, the GRP94 inhibitor is NVP-AUY922 or a pharmaceutically acceptable salt, isomer or solvate thereof.

Preferably, the GRP94 inhibitors are for use in the treatment or prevention of brain metastasis from cancer, preferably brain metastasis from lung cancer or breast cancer, in patients overexpressing the GRP94 protein.

HDAC inhibitor compounds

In another aspect, the invention is directed to a HDAC inhibitor compound for use in the prevention or treatment of brain metastasis from cancer, preferably brain metastasis from lung cancer or brain metastasis from breast cancer.

The term "HDAC inhibitor" as used herein refers to a compound that is capable of inhibiting the deacetylation of histones in vivo, in vitro or both. As such, a HDAC inhibitor inhibits the activity of at least one histone deacetylase. As a result of inhibiting the deacetylation of at least one histone, an increase in acetylated histone occurs and accumulation of acetylated histone is a suitable biological marker for assessing the activity of HDAC inhibitors. Therefore, procedures which can assay for the accumulation of acetylated histones can be used to determine the HDAC inhibitory activity of compounds of interest.

Non-limiting examples of HDAC inhibitors include compounds of the following classes: hydroxamic acids (e.g., trichostatin A, vorinostat or suberoylanilide hydroxamic acid, belinostat, panobinostat, abexinostat, pracinostat, givinostat and quisinostat), cyclic tetrapeptides (e.g. trapoxin, apicidin and depsipeptide), benzamides (e.g. entinostat, CI-994 and mocetinostat), electrophilic ketones (e.g. trifluoromethyl o ketones and oketoamides) and aliphatic acids (e.g. valproic acid, sodium valproate, butyric acid, sodium butyrate, pivanex, sodium phenylbutyrate, isovalerate, butrymide, isobutyramide, 3-bromopropionate and tributyrin).

In a particular embodiment, the HDAC inhibitor is an aliphatic acid-class HDAC inhibitor, for example valproic acid, sodium valproate, butyric acid, sodium butyrate, pivanex, sodium phenylbutyrate, isovalerate, butrymide, isobutyramide, 3- bromopropionate and tributyrin, or a pharmaceutically acceptable salt or solvate thereof. Preferably, the HDAC is selected from valproic acid and sodium valproate. In an embodiment, the HDAC is valproic acid. In another embodiment, the HDAC inhibitor is sodium valproate.

In a particular embodiment, the primary tumor responsible for brain metastasis is selected from lung cancer, breast cancer, bladder cancer, prostate cancer, kidney cancer, pancreatic cancer, colon cancer, skin cancer, sarcoma and lymphoma. Preferably, it is selected from breast cancer and lung cancer.

In a particular embodiment, the invention is directed to a compound selected from a compound (I) as defined above, or a pharmaceutically acceptable salt, isomer or solvate thereof, a GRP94 inhibitor compound and a HDAC inhibitor compound for use in the prevention or treatment of brain metastasis from lung cancer or brain metastasis from breast cancer. Preferred embodiments for the compound of formula (I), GRP94 inhibitor compound and HDAC inhibitor compound are as defined herein.

In a particular embodiment, the invention is directed to a compound (I) as defined above, or a pharmaceutically acceptable salt, isomer or solvate thereof, for use in the prevention or treatment of brain metastasis from breast cancer.

In an embodiment, the invention is directed to a GRP94 inhibitor compound as defined above for use in the prevention or treatment of brain metastasis from breast cancer.

In an embodiment, the invention is directed to a HDAC inhibitor compound as defined above for use in the prevention or treatment of brain metastasis from breast cancer.

In a particular embodiment, the breast cancer is negative for at least one of: estrogen receptor (ER), progesterone receptor (PR) or human epidermal growth factor receptor 2 (HER2).

In an embodiment, the breast cancer is ER-negative breast cancer. In an embodiment, the breast cancer is PR-negative breast cancer. In an embodiment, the breast cancer is HER2-negative breast cancer. In an embodiment, the breast cancer is ER-negative, HER2-negative and PR-negative (triple negative breast cancer).

In another embodiment, the breast cancer is positive for at least one of: estrogen receptor (ER), progesterone receptor (PR) or human epidermal growth factor receptor 2 (HER2). In an embodiment, the breast cancer is ER-positive breast cancer. In an embodiment, the breast cancer is PR-positive breast cancer. In an embodiment, the breast cancer is HER2-positive breast cancer. In an embodiment, breast cancer is both ER-positive and PR-positive.

In a preferred embodiment, breast cancer is selected from HER2-positive breast cancer and ER-negative, HER2-negative and PR-negative breast cancer. In an embodiment, breast cancer is HER2-positive breast. In an embodiment, breast cancer is ER-negative, HER2-negative and PR-negative breast cancer.

The expressions "HER2-negative", "ER-negative" and "PR-negative" refer to specific subtype of breast cancer that does not express the genes for HER2, estrogen receptor (ER) or progesterone receptor (PR), respectively.

The expressions "HER2-positive", "ER-positive" and "PR-positive" mean characterized by overexpression of HER2, ER and PR protein, respectively. Such overexpression can be determined by methods well known in the art, such as immunohistochemistry, fluorescence in situ hybridization or ligand binding assays.

In a particular embodiment, the invention is directed to a compound (I) as defined above, or a pharmaceutically acceptable salt, isomer or solvate thereof, for use in the prevention or treatment of brain metastasis from lung cancer, including non-small cell lung cancer and small cell lung cancer.

In an embodiment, the invention is directed to a GRP94 inhibitor compound as defined above for use in the prevention or treatment of brain metastasis from lung cancer, including non-small cell lung cancer and small cell lung cancer.

In an embodiment, the invention is directed to a HDAC inhibitor compound as defined above for use in the prevention or treatment of brain metastasis from lung cancer, including non-small cell lung cancer and small cell lung cancer.

Pharmaceutically acceptable salts can be synthesized from the parent compound by conventional methods. For example, such salts can be prepared by reacting the free base form of the compound with a stoichiometric amount of the appropriate acid in water, in an organic solvent or in a mixture of the two. Examples of acid addition salts include mineral acid addition salts such as, for example, hydrochloride, hydrobromide, hydroiodide, sulphate, nitrate, phosphate, and organic acid addition salts such as, for example, acetate, trifluoroacetate, maleate, fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate, methanesulphonate and p-toluenesulphonate. The compounds of the invention can be in free form or in solvate form (for example, hydrates, alcoholates, etc.), both forms being included within the scope of the present invention. Solvation methods are generally well known in the state of the art.

Compounds referred to herein may have asymmetric centers and therefore exist in different enantiomeric forms. All optical isomers and stereoisomers of the compounds referred to herein, and mixtures thereof, are considered within the scope of the present invention. Thus any given compound referred to herein is intended to represent any one of a racemate, one or more enantiomeric forms, one or more diastereomeric forms, and mixtures thereof. The single isomers and mixtures of isomers fall within the scope of the present invention.

The term "pharmaceutically acceptable" relates to molecular entities and compositions being physiologically tolerable and normally not causing an allergic reaction or similar adverse reaction, such as gastric discomfort, dizziness and the like, when they are administered to a human being. Preferably, as used herein, the term "pharmaceutically acceptable" means approved by a governmental regulatory agency or listed in the US pharmacopoeia or another generally recognized pharmacopoeia for use in animals, and more particularly in humans.

Pharmaceutical compositions

In another aspect, the invention is directed to a pharmaceutical composition comprising a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, isomer or solvate thereof, a GRP94 inhibitor and a pharmaceutically acceptable carrier.

Preferred embodiments for the compound of formula (I) and for the GRP94 inhibitor are as defined herein.

In a particular embodiment, the compound of formula (I) is selected from thalidomide, lenalidomide and pomalidomide, or a pharmaceutically acceptable salt, isomer or solvate thereof, and the GRP94 inhibitor is selected from 17-AAG and NVP- AUY922, or a pharmaceutically acceptable salt, isomer or solvate thereof. In a further embodiment, the compound of formula (I) is lenalidomide or a pharmaceutically acceptable salt, isomer or solvate thereof, and the GRP94 inhibitor is NVP-AUY922, or a pharmaceutically acceptable salt, isomer or solvate thereof.

In another aspect, the invention is directed to a pharmaceutical composition as defined above, for use in therapy. In a further aspect, the invention is directed to a pharmaceutical composition as defined above, for use in the prevention or treatment of brain metastasis from cancer, preferably brain metastasis from breast cancer or brain metastasis from lung cancer.

Further aspects of the invention refer to the use of a pharmaceutical composition as defined above in the preparation of a medicament and to the use of a pharmaceutical composition as defined above in the preparation of a medicament for the prevention or treatment of brain metastasis from cancer, preferably brain metastasis from breast cancer or brain metastasis from lung cancer.

The term "pharmaceutically acceptable carrier" in the compositions according to the present invention refers to pharmaceutically acceptable solvents, suspending agents or vehicles for delivering a compound of the present invention and which are acceptable from a toxicological viewpoint. The number and the nature of the pharmaceutically acceptable carriers depend on the desired administration form. Suitable pharmaceutical carriers are known by the person skilled in the art, and they and the methods of formulating the compositions can be found in standard references (e.g. "Remington: The Science and Practice of Pharmacy", 20th edition (2003) Genaro A.R., ed., Lippincott Williams & Wilkins, Philadelphia, US).

As used herein, the terms "treatment" or "treating" refer to both therapeutic measures and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) an undesired physiological change or disorder. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. "Treatment" can also mean prolonging survival as compared to expected survival if not receiving treatment.

The terms "prevention" or "preventing" as used herein mean that the compounds are useful when administered to a patient who has not been diagnosed as possibly having the disorder or disease at the time of administration, but who would normally be expected to develop the disorder or disease or be at increased risk for the disorder or disease. The compounds of the invention will slow the development of the disorder or disease symptoms, delay the onset of the disorder or disease, or prevent the individual from developing the disorder or disease at all. As used herein, the expression "effective amount" refers to the sufficient amount of the compound to provide the desired effect and will generally be determined by, among other causes, the characteristics of the compound itself and the therapeutic effect to be achieved. It will also depend on the subject to be treated, the severity of the disease suffered by said subject, the chosen dosage form, administration route, etc. For this reason, the doses mentioned in this invention must be considered only as guides for the person skilled in the art, and he must adjust the doses depending on the aforementioned variables. However, the compound of the invention can be administered one or more times a day, for example, 1 , 2, 3 or 4 times a day in a typical total daily amount comprised between 0.1 and 30 mg/kg of body mass/day, preferably between 1 and 10 mg/kg of body mass/day. In an embodiment, the effective amount produces at least one therapeutic effect selected from the group consisting of reduction in metastasis, complete remission, partial remission, stable disease, increase in overall response rate, or a pathological complete response.

The compounds and compositions described herein may be administered by any suitable administration route, such as, but not limited to, parenteral, oral, topical, nasal, rectal route. In a particular embodiment, the compounds and compositions described herein are administered by parenteral route, e.g. by intravenous, intraperitoneal, subcutaneous, intradermal, intramuscular, intrathecal or epidural administration. In a further embodiment, it is administered intravenously.

Illustrative examples of dosage forms for administration by the oral route include tablets, capsules, granulate, solutions, suspensions, etc., and can contain the conventional excipients, such as binders, diluents, disintegrants, lubricants, wetting agents, etc., and can be prepared by conventional methods. The pharmaceutical compositions can also be adapted for their parenteral administration, in the form of, for example, solutions, suspensions, emulsions or lyophilized products, etc and can include the suitable excipients, such as buffers, surfactants, anti-oxidants, etc. In any case, the excipients will be chosen according to the selected pharmaceutical dosage form.

Typically, the compounds of the invention will be given in the range of 0.1 to 30 mg/kg of body mass per day, preferably in the range of 1 to 10 mg/kg of body mass per day.

As used herein, the term "patient" means any mammal, preferably a human. Combination Therapy

In an embodiment, the method and uses provided herein may further comprise another anti-cancer therapy including, but not limited to, surgery, radiation therapy, chemotherapy, or a combination thereof.

In a particular embodiment, the additional anti-cancer therapy is chemotherapy including administration of at least one other chemotherapeutic agent. The administration in combination with other chemotherapeutic agent may be in a unitary pharmaceutical composition comprising both compounds or in separate pharmaceutical compositions each including one of the compounds. The combination may be administered separately in a sequential manner wherein one agent is administered first and the other second or vice versa. Such sequential administration may be close in time or remote in time.

In an embodiment, the compound of the invention is administered simultaneously, separately or sequentially with at least one other chemotherapeutic agent. Preferably, said other chemotherapeutic agent is selected from the group consisting of taxanes, antimetabolites, platinum-based agents, alkylating agents, tyrosine kinase inhibitors, anthracycline antibiotics, vinca alkaloids, topoisomerase I or II inhibitors, proteasome inhibitors and macrolides.

In a particular embodiment, a compound of formula (I), or a pharmaceutically acceptable salt, isomer or solvate thereof, as defined herein is administered simultaneously, separately or sequentially with at least one other chemotherapeutic agent, for use in the prevention or treatment of brain metastasis from cancer, preferably brain metastasis from breast cancer or brain metastasis from lung cancer.

In an embodiment, a compound of formula (I), or a pharmaceutically acceptable salt, isomer or solvate thereof, as defined herein is administered simultaneously, separately or sequentially with docetaxel, for use in the prevention or treatment of brain metastasis from cancer, preferably brain metastasis from breast cancer or brain metastasis from lung cancer. Preferred embodiments of the compound of formula (I) are as defined herein. In a particular embodiment, compound of formula (I) is selected from a compound wherein R¹ is NH₂, R² is H, and n is selected from 0 and 1. In a preferred embodiment, the compound of formula (I) is selected from thalidomide, lenalidomide and pomalidomide, or a pharmaceutically acceptable salt, isomer or solvate thereof. More preferably, the compound of formula (I) is lenalidomide, or a pharmaceutically acceptable salt, isomer or solvate thereof. The compound of formula (I) and docetaxel may be provided as a single medicament or as separate medicaments for administration at the same time or at different times. Preferably, they are provided as separate medicaments for administration at different times. When administered separately and at different times, either the compound of formula (I) or docetaxel may be administered first. In addition, both drugs can be administered in the same day or at different days, following the same schedule or different schedules during the treatment cycle and through the same or different administration route.

In another embodiment, a compound of formula (I), or a pharmaceutically acceptable salt, isomer or solvate thereof, as defined herein is administered simultaneously, separately or sequentially with a GRP94 inhibitor, for use in the prevention or treatment of brain metastasis from cancer, preferably brain metastasis from breast cancer or brain metastasis from lung cancer.

In an embodiment, the invention refers to a combination comprising a compound of formula (I), or a pharmaceutically acceptable salt, isomer or solvate thereof, and a GRP94 inhibitor, for simultaneous, separate or sequential use in therapy.

In another embodiment, the invention refers to a combination comprising a compound of formula (I), or a pharmaceutically acceptable salt, isomer or solvate thereof, and a GRP94 inhibitor, for simultaneous, separate or sequential use in the prevention or treatment of brain metastasis from cancer, preferably brain metastasis from breast cancer or brain metastasis from lung cancer.

Preferred embodiments of the compound of formula (I) and the GRP94 inhibitor are as defined herein. In a particular embodiment, compound of formula (I) is selected from a compound wherein R¹ is NH₂, R² is H, and n is selected from 0 and 1 , and the GRP94 inhibitor is selected from 17-AAG and NVP-AUY922, or a pharmaceutically acceptable salt, isomer or solvate thereof. In a preferred embodiment, the compound of formula (I) is selected from thalidomide, lenalidomide and pomalidomide, or a pharmaceutically acceptable salt, isomer or solvate thereof, and the GRP94 inhibitor is selected from 17-AAG and NVP-AUY922, or a pharmaceutically acceptable salt, isomer or solvate thereof. More preferably, the compound of formula (I) is lenalidomide, or a pharmaceutically acceptable salt, isomer or solvate thereof, and the GRP94 inhibitor is NVP-AUY922, or a pharmaceutically acceptable salt, isomer or solvate thereof. The above mentioned combinations of the compound of formula (I) and another chemotherapeutic agent may provide a synergetic effect in the prevention or treatment of brain metastasis from cancer, preferably brain metastasis from breast cancer or brain metastasis from lung cancer, in particular, the combination of lenalidomide or a pharmaceutically acceptable salt, isomer or solvate thereof and docetaxel. Also, the above mentioned combinations of the compound of formula (I) and the GRP94 inhibitor may provide a synergetic effect in the prevention or treatment of brain metastasis from cancer, preferably brain metastasis from breast cancer or brain metastasis from lung cancer, in particular, the combination of lenalidomide or a pharmaceutically acceptable salt, isomer or solvate thereof and NVP-AUY922, or a pharmaceutically acceptable salt, isomer or solvate thereof.

The compound of formula (I) and the GRP94 inhibitor may be provided as a single medicament or as separate medicaments for administration at the same time or at different times. Preferably, they are provided as separate medicaments for administration at different times. When administered separately and at different times, either the compound of formula (I) or the GRP94 inhibitor may be administered first. In addition, both drugs can be administered in the same day or at different days, following the same schedule or different schedules during the treatment cycle and through the same or different administration route.

In another particular embodiment, the additional anti-cancer therapy is radiotherapy.

The term "combination" as used throughout the specification, is meant to encompass the administration of the therapeutic agents in the same or separate pharmaceutical formulations, and at the same time or at different times.

Simultaneous administration as used herein means that a first compound and a second compound are administered with a time separation of no more than about 20 minutes, such as no more than about any of 15, 10, 5 or 1 minutes or administered at the same time. Both compounds may be contained in the same composition or in separate compositions.

Sequential administration as used herein means that a first compound and a second compound are administered with a time separation of more than about 20 minutes, such as more than about 30, 40, 50, 60, 120 or more minutes. Both compounds are contained in separate compositions, which may be contained in the same or different packages or kits. Separate administration means that one component may be administered first and the second component is administered thereafter, with no limitation as to the duration of time. Accordingly the term "separate administration" encompasses both the time limits of simultaneous and sequential administration.

In a further aspect, the invention refers to a kit for administration of a compound of formula (I), or a pharmaceutically acceptable salt, isomer or solvate thereof, as defined herein and a GRP94 inhibitor. Kits may further comprise packaging and/or printed instructions for use of the combination.

In another aspect, the invention refers to a kit for administration of a compound of formula (I), or a pharmaceutically acceptable salt, isomer or solvate thereof, as defined herein and docetaxel. Kits may further comprise packaging and/or printed instructions for use of the combination.

The following examples illustrate the invention and must not be considered in a limiting sense thereof.

EXAMPLES

Example 1 : Cytotoxicity assays

MDA-MB 435 cell cultures(435-P) and 435-Br1 cells, established from brain metastasis in nude mice, with ability to metastasize in brain, were maintained under standard conditions (Schackert, G.; Price, J. E.; Bucana, C. D.; Fidler, I. J. Int. J. Cance/1989, 44 (5), 892-897; Zhang, R. D.; Fidler, I. J.; Price, J. E. Invasion Metastasis 1991 , 11 (4), 204-215; Fernandez, Y.; Espana, L; Manas, S.; Fabra, A.; Sierra, A. Cell Death Differ. 2000,7 (4), 350-359). The 435-Br1 obtained from the brain when mice were injected for the first time in the left ventricle are named V1 cells, and those obtained after the fifth injection are named V5 cells. This is a procedure to select the most organspecific metastatic variant. V5 cells were injected in the internal carotid artery and a new variant V5CA1 was obtained (Martinez et al., Development of a preclinical therapeutic model of human brain metastasis with chemotherapy. IJMS Manuscript ID: ijms-25852 in press). Additionally, brain metastasis variants from human breast cancer provided through the METABRE consortium: MDA-MB 361 (Laboratoire d'Oncogenetique, Centre Rene Huguenin, Saint-Cloud, France) were also used.

The results obtained after 72 h of treatment with 17-AAG are shown in Figure 1. Survival is expressed as the percentage of cells with regard to the control cells without treatment. Example 2: In vivo/in vitro characterization of NVP action on MDA-MB 435 parental breast cancer cells (435/P) and the brains metastatic variant 435/Br1

Western blotting.- The selected cell lines 435/P, 435 B , Br1V5 and B V5CA1 derived from in vivo/in vitro successive rounds to obtain high brain metastatic cells (Martinez-Aranda A, Hernandez V, Picon C, Modolell I, Sierra A. Development of a preclinical therapeutic model of human brain metastasis with chemoradiotherapy. Int J Mol Sci. 2013 Apr 16;14(4):8306-27) were lysed in a 1 % SDS (v/v) extraction buffer containing an anti-protease cocktail (Roche, Vilvoorde, Belgium). Protein concentrations were determined using the Bradford assay (MicroBCA, Pierce, Belgium). After resolution by SDS-PAGE, electrophoresed proteins were transferred to polyvinylidene fluoride (PVDF) membranes that were blocked and probed with the following antibodies: GRP94 (1/1000, Sta Cruz) and a-tubulin (1/10000, Sigma), with the corresponding peroxidase-conjugated secondary antibody at 1 :2000: Peroxidase conjugated anti-rabbit secondary Ab (Amersham), Peroxidase conjugated Antimouse secondary Ab (Pierce, Perbio Science Ltd., Cheshire, U.K). Immunoreactive bands were quantified using a VersaDoc™ (Bio-Rad) Imaging System using the Super Signal west-Pico (Pierce). MWs were established with See Blue Plus2 prestained Standford (Invitrogen, San Diego, CA). Results are shown in Figure 2A.

Cytotoxic assay.- To check NVP cytotoxicity we treated 6x10³ Br1V5 cells/well with 5- 50 nM of NVP in DMSO 100% during 72 h. MTT survival assay was performed with 1 %DMSO to assess the IC50 of the NVP, which exerted a cytotoxic effect with a IC50 at 50 nM. Results are shown in Figure 2B.

Example 3: In vivo treatment of subcutaneous xenograft of brain metastasis from lung carcinoma- Experimental models.- Athymic Nude-Foxn1 nu mice of 22-28 gr weight (Charles-River Laboratories (Wilmington, MA) were used. Small pieces of 2 mm³ from brain metastases biopsies were implanted subcutaneously on the back or intra mammary fat path, respectively. After 10 days when the engraftment was growing we started the treatment with the drug or with the vehicle. Daily control of weight and tumor volume was used to follow the treatment. Tumors were explored in paraffin sections by hematoxylin-eosin stain and specific antibodies.

Drugs used and protocols of treatment.- Mice were treated with: Docetaxel (TXT - comparative assay) 15 mg/Kg/day, every 4 days, for two weeks; with Lenalidomide (LND) 50 mg/Kg/day, 5 days a week for two weeks; GRP94 inhibitors (NVP-AUY922 ) 30mg/Kg/day, 3 doses per week, for two weeks and 17AAG 40 mg/Kg/d in 20% Cremophor, i.p. 5 days a week for two weeks; Valproic Acid 100mg/Kg/day5 days a week for two weeks. All of them were administered intra peritoneal {i.p.). Controls were treated with the vehicle: physiologic serum 50% DMSO. In the case of combinations, the treatments were administered at the same doses. No secondary effects were detected.

The results obtained are shown in Figures 3-5. Treatment with the compounds of the invention slow tumours growth compared to the control treated tumours. Comparable or improved results over treatment with taxotere (standard treatment) were observed.

Claims

1. A compound of formula (I)

wherein

X is selected from -CH₂- and -C(O)-;

R² is selected from H and CrC₆ alkyl; and

n is an integer selected from 0, 1 , 2, 3 and 4;

or a pharmaceutically acceptable salt, isomer or solvate thereof,

for use in the prevention or treatment of brain metastasis from cancer.

2. A compound for use according to claim 1 , wherein R¹ is NR'R" and R' and R" are independently selected from H, and Ci-C₆ alkyl.

3. A compound for use according to claim 2, wherein R¹ is NH₂ and n is 0 or 1.

4. A compound for use according to any of claim 1 to 3, wherein R² is H.

5. A compound for use according to any of claim 1 to 4, wherein the compound of formula (I) is selected from thalidomide, lenalidomide and pomalidomide, or a pharmaceutically acceptable salt, isomer or solvate thereof.

6. A compound for use according to any of claims 1 to 5, wherein compound of formula (I) is administered simultaneously, separately or sequentially with a compound selected from GRP94 inhibitor and docetaxel.

7. A GRP94 inhibitor for use in the prevention or treatment of brain metastasis from cancer.

8. A GRP94 inhibitor for use according to claim 7, which is selected from 17-AAG and NVP-AUY922, or a pharmaceutically acceptable salt, isomer or solvate thereof.

9. A HDAC inhibitor for use in the prevention or treatment of brain metastasis from cancer.

10. A HDAC inhibitor for use according to claim 9, which is selected from valproic acid and sodium valproate, or a pharmaceutically acceptable salt, isomer or solvate thereof.

1 1 . A compound for use according to any of claims 1 to 10, wherein the brain metastasis from cancer is selected from brain metastasis form lung cancer and brain metastasis from breast cancer.

12. A compound for use according to claim 1 1 , wherein the breast cancer is HR2- positive or triple negative breast cancer.

13. A pharmaceutical composition comprising a compound of formula (I) as defined in any of claims 1 to 5, a GRP94 inhibitor and a pharmaceutically acceptable carrier.

14. A pharmaceutical composition according to claim 13, wherein the compound of formula (I) is lenalidomide, or a pharmaceutically acceptable salt, isomer or solvate thereof, and the GRP94 inhibitor is NVP-AUY922, or a pharmaceutically acceptable salt, isomer or solvate thereof.

15. A pharmaceutical composition according to any of claims 13 to 14, for use in the prevention or treatment of brain metastasis from cancer.