ITPA20070034A1 - GEMINAL BIPHOSPHONATES, THEIR PREPARATION AND THEIR USE IN THE ONCOLOGICAL FIELD. - Google Patents

Description

DESCRIPTION

Attached to a patent application for INDUSTRIAL INVENTION having as its title

"GEMINAL BIPHOSPHONATES, THEIR PREPARATION AND THEIR USE IN THE ONCOLOGICAL FIELD"

The present invention relates to geminal bisphosphonic compounds (or geminal bisphosphonates), having a structure that can be correlated with that of zoledronic acid, a method for their preparation, their pharmaceutical compositions and their use in oncology, for the treatment of tumor pathologies.

In particular, the present invention relates to the use of said bisphosphonates for the preparation of pharmaceutical compositions for the treatment of tumor pathologies that are not correlated, neither correlated nor dependent on calcium.

Geminal bisphosphonates are compounds having a chemical structure similar to that of endogenous pyrophosphate: in fact, in said compounds a carbon atom replaces the central oxygen atom. This replacement makes these compounds resistant to hydrolysis (that is, more stable inside the organism) and also allows the insertion of two additional chains of variable structure on said carbon atom, which can influence the pharmacological activity of the molecules. Almost always one of these two chains contains a hydroxyl group which allows the compound to have a high affinity for calcium and for the mineral part of the bone. presence of free hydroxyl groups (acids) on the two phosphoric acid residues.

Thanks to their high affinity for calcium and for the mineral part of the BONE, the above mentioned geminal bisphosphonates are well known, and as such used in therapy, for their ability to inhibit bone resorption,

Consequently, the pharmacological activity of said derivatives is, known to be, closely correlated and dependent on calcium and the mineral part of the bone.

Among said bisphosphonates, it is possible to mention, by way of example, compounds selected from the group comprising: zoledronic acid, alendronic, ibandronic, etidronic, pamidronic acid.

Due to the strong affinity for calcium described above, some of said phosphonates have also been found to have a certain antitumor activity (for example, anti-myeloma) in those specific tumor pathologies which are correlated and / or correlatable and or dependent on calcium.

However, it would be useful to have available compounds which are structurally belonging to the group of geminal bisphosphonates, but which are characterized by a significant specific activity against those tumor pathologies which are unrelated and or unrelated and / or not dependent on calcium.

Compounds meeting the requirements described above are not known. In particular, there are no known compounds with the aforementioned specific activity against those tumor pathologies that are unrelated and / or unrelated and / or not dependent on calcium, whose structure is attributable to that of zoledronic acid.

Therefore, the need remains to have available compounds characterized by the ability to act significantly in the specific treatment of tumor pathologies that are unrelated and / or unrelated and / or not dependent on calcium. The purpose of the present invention is to respond to the above need.

This object and still others, which will become clear from the detailed description that follows, have been achieved by the Applicant, who has unexpectedly found that a suitable group of compounds, having a structure referable to that of zoledronic acid, is able to give a adequate response to the need above highlight it.

An object of the present invention is the use of said above compounds for the preparation of a medicament (i.e., a pharmaceutical composition) for the treatment of tumor pathologies (and / or pathologies referable to them or derivable from them) which are unrelated and / or unrelated and / or not dependent on calcium, as reported in the attached independent claim.

Furthermore, an object of the present invention is the use of said compounds for the preparation of a pharmaceutical composition for the treatment of related pathologies or due to abnormal angiogenesis, as reported in the attached claims.

A further object of the present invention are the new geminal bis-phosphonate compounds endowed with the above activity and their pharmaceutical compositions, as reported in the attached claims.

Preferred embodiments of the present invention are reported in the attached dependent claims.

The present invention is illustrated in detail in the following description. Said invention is further illustrated also with the aid of the enclosed Tables 1 to X

Table 1 shows in graph the inhibitory effect exerted, at different dosages, of one of the preferred compounds of the present invention (indicated, in acronym, as MM32 and described later in the description) on the growth of different cell lines (respectively, H.UT78: IC562 : CEM and CEM VBL300).

Table 2 graphs the inhibitory effect exerted (at the same dosages reported in Table 1 for MM32) by zoledronic acid on the growth of the same cell lines as in Table 1 (respectively, HUT78; K562; CEM and CEM VBL300).

Table 3 shows in block graph the percentage of apoptosis induced by MM32 at a concentration of 100 μΜ. compared to that induced by the zoledronic phase, respectively, on the cell lines HUT78, K562, CEM and CEM VBL300

Table 4 shows in block graph the percentage of apoptosis induced by the combination imatinib zoledronic acid and, respectively, by the combination imatinib MM32, compared to that induced by imatinib alone, on the K562 cell line; the imatinib compound is used at a concentration of 0.25 μΜ; both zoledronic acid and MM32 are used at a concentration of 30 μΜ.

Table 5 illustrates in block graph the low cytotoxicity of MM32. compared to that of zoledronic acid, on normal hematopoietic stem cells of the CFU-CM line: the cytotoxic effect was determined, - for both compounds, a. concentrations ranging from 20 to 100 μΜ,

The use of one or more compounds of formula fi) is a preferred object of the invention:

their enantiomers, diastereomers, or their respective mixtures, and / or their pharmaceutically compatible salts for the preparation of a pharmaceutical composition for the treatment of at least one tumor pathology that is unrelated and / or uncorrectable and / or not dependent on calcium .

Preferably, said compounds of formula (!) Are selected from the group in which:

their enantiomers. diastereomers. and / or their mixtures, and / or their pharmaceutically compatible salts.

More preferably, said compounds of formula (T) are selected from the group in which: R, is H. C: H5;

Y is H;

X is CHg

R3 is a group of formula:

their enantiomers, diastereomers, and / or their mixtures, and / or their pharmaceutically compatible salts,

Basically. The compounds of formula (I) are structural analogues of zoledronic acid, in which the group OH. on the central carbon of the molecule has been eliminated, in order to decrease the affinity of the compound for calcium. Furthermore, the imidazole residue of zoledronic acid has been variously replaced with heterocyclic residues, aromatic and otherwise. Said residues are preferably selected from the group comprising: benzimidazole, benzopyrrole, indole, indazoium, pyrazole, pyridine * pyridazine. pyrazine, pyrrolidine, piperazine, piperidine, imidazopyridazine. imidazopyrazma. imidazopyridine, purines. ; Quite unexpectedly, the compounds of formula (I) proved to be very active in the active yδ T-lymphocytes. In fact, compared, for example, to zoledronic acid, they have been shown to have an activating action on average at least 100 times higher, up to 1000 times higher and even more. Some of the preferred compounds have even shown to possess an activating activity against yδ T-lymphocytes comparable to that exhibited by the natural phosphoantigen (E) -4-hydroxy-3-methyl-buL-2-enyl-pyrophosphate. Equally unexpectedly, said compounds have also been shown to possess a high cytotoxic and apoptotic activity, thus proposing themselves as excellent candidates as anticancer agents. in a particularly unexpected way, the compounds of formula (I) have proved particularly useful for the specific treatment of those tumor pathologies which are not correlated and / or not correctable and / or not dependent on calcium. In a preferred embodiment of the invention, said compounds have proved to be usable for the treatment of at least one tumor pathology sensitive to the action of yδ T-lymphocyte activating substances. ; In another preferred embodiment, said compounds can be used for the treatment of at least one tumor pathology sensitive to the action of substances with cytotoxic activity. ; In a further preferred embodiment, said compounds have proved to be usable for the treatment of at least one tumor pathology sensitive to the action of substances with apoptotic activity. With reference to the above, by way of absolutely non-limiting example, one or more compounds of formula (J) of the present invention, as well as their astereoisomers enantiomers. and / or their mixtures, and / or their pharmaceutically compatible salts, are usable for the preparation of a pharmaceutical composition for the treatment of at least one non-related and / or non-correlable and / or calcium-dependent tumor pathology, in which said tumor pathology is chosen from the group comprising: sarcoma, carcinoma, carcinoid renal carcinoma, colon cancer, breast cancer, bone cancer, neuroendocrine cancer, prostate cancer, leukemia, Ber-Abl positive leukemia. my lagena chronic leukemia, lymphoid leukemia, myeloid leukemia, megakaryocytic leukemia. non-Hodgkin's lymphomas, melanoma, Hodgkin's disease. tumor pathologies sensitive to immunotherapies, gastrointestinal stremal tumors (GISTs), the compounds of formula {1} have also shown themselves to be endowed with activity against referable pathologies and / or due to the tumoral ones described above, for example, with abnormal angiogenesis. Consequently, the use of said compounds for the preparation of a pharmaceutical composition for the treatment of a pathology correlated and / or correctable to abnormal angiogenesis is a further object of the present invention. By way of example, delta pathology correlated and / or correctable to abnormal angiogenesis is chosen from the group comprising: arthritis, tumors responding to angiogenic activity, metastatic spread, diabetic retinopathy, psoriasis, chronic inflammation, atherosclerosis. In a particularly preferred embodiment of the invention, said one or more compounds of formula (I) is formulated in combination with at least one compound with antiblastic activity. Said at least one compound endowed with antiblastic activity is. preferably, selected from the group comprising: alkylating agents, topoi somerase inhibitors, melt poisons, intercalating agents, antimetabolites, products of natural origin, such as. Vinca alkaloids, epipodophilloioxins, antibiotics, enzymes, Texans, anti-cancer vaccines. The preparation of the compounds of formula (I) is illustrated in the following Synthesis Scheme 1, with specific reference to the most preferred compounds. However, the same conditions are easily applicable by the skilled person also to the preparation of the other compounds of formula (I). In particular, said Scheme 1 summarizes the method used for the preparation of different classes of bisphosphonates of formula (I), in which the respective geminal bisphosphonates (i.e. the bisphosphonic esters) are prepared from the corresponding methylen-bisphosphonate by treatment with the appropriate NH -derivative, as shown in the diagram, and with TBD (ie 1,5,7-trìazabìciclo [4 * 4.0] dec-5-ene) as catalyst, in the presence or not of a suitable solvent.

Alternatively, the geminal bisphosphonates of the invention are prepared from the corresponding methylene bisphosphonate by treatment with the appropriate NH-derivative at 140-170QC per top 10 - 20 min in a microwave oven (MW), in turn, the corresponding bisphosphonic acids are prepared by the additive treatment of the respective esters mentioned above.

Preparation of the tetraethyl ester of the ethenylidene bisphosphonic acid of formula (1) of Scheme 1

1 equiv. of paraformaldehyde and I equiv. of diethylamine are mixed in methanol (20, 30 ml) and heated until a clear solution is obtained. Then, 1. equivalent of tetractyl-methylene-bisphosphonate is added and the mixture is refluxed for 18h. The resulting crude is concentrated under vacuum, taken up with methanol and reconcentrated under vacuum; after which, it is taken up with toluene and again concentrated under vacuum. The residue is solubilized in toluene, treated with a catalytic quantity of pTSA (p-toluene sulfuric acid), then it is refluxed through a Dean-Stark trap for 18h. The residue is then concentrated under vacuum, dissolved in chloroform, washed twice with water, dried over anhydrous sodium sulphate and reconcentrated under vacuum. The residue is a yellow oil which is used as such, without further purification. Yield: 90%.

Preparation of the tetractyl bisphosphonates of formula (2 a-1) of Scheme 1 Method A): a solution of one of the compounds a-1 (1.1 equiv.) And 1 (1 equiv.) In T1IF (tetra! in a sealed vial and heat it to 150-160 ° C for 15 min in a microwave oven (MW), After cooling to room temperature (20-25 ° C), the solvent is evaporated under vacuum and the residue is dissolved in water and extract with dichloromethane. The organic phases are combined, dried over anhydrous sodium sulphate, filtered and evaporated. The crude residue is purified by flash chromatography on silica with dichlorometha-methanol (98: 2 / v: v). Yield: 50-90%.

Method B); a mixture of a compound a-1 (1.1 equiv.) and 1 (I equiv.) in THF is refluxed under stirring for 4h, after which the solvent is evaporated under vacuum. The crude residue is solubilized in water and extracted with dichloromethane; the organic phases are combined, dried over anhydrous sodium sulphate, filtered and evaporated. The residue is purified by flash chromatography on silica with dichloromethane-methanol (98: 2 / v; v), Yield: 40-70%.

Yellow oil. Yield: 70%

Preparation of the eternal bisphosphonic aikiliden acids of formula (3 a-t) of Scheme 1

Method A): the solution of one of the compounds 2 a-1 in concentrated HCI is kept under reflux under stirring for 3h. then, the solvent is evaporated under vacuum. The solid residue is crystallized from H2Ο-methanol to give the corresponding compound 3 a-1 in the form of a white or yellow solid.

Yield: 60-90%.

Method B): to a solution, externally cooled with an ice bath, of one of the compounds 2 a-1 (1 equiv.) In CCl4 iodotrimelilsilane (4.2 equiv.) Is added drop by drop and the resulting mixture is stirred for three hours keeping the temperature below 5 ° C. After that. the reaction mixture is stopped by adding methanol and the precipitate formed is collected and crystallized from H2O -methanol to give the corresponding compound 3 a-1 in the form of a white or yellow solid.

Yield: 40-80%.

With the method described above, for example, the compounds were prepared:

Claims (15)

CLAIMS Use of one or more compounds of formula (T): in which: A and Z. independently of each other "are C. CH, N, NH, S; Ri and Ri. equal or different from each other, they are H. CH .. C <, Hs, GOL, ΝΗ:>, NOs, CNfCOGLI, halogen, 28 in which: R is C i - C4o alkyl at le, their enant iomers, diastereoi somers, or their respective mixtures, and / or their pharmaceutically compatible salts for the preparation of a pharmaceutical composition for the treatment of at least one tumor pathology that is unrelated and / or non-correctable and / or non-dependent from! soccer. 2. The use according to claim I wherein some compounds of formula (1) are selected from! group in which " toro enanliomeri, diastereoi someri, and / or their mixtures, and / or their compatible pharmacological salts, 3, The use according to claim 1 or 2, wherein said compounds of formula (1) are selected from the group in cut: their enantiomers, diastereomers. and / or their mixtures, and / or their pharmaceutically compatible salts The use according to any one of the preceding claims, in which said at least one tumor pathology is sensitive to the action of substances activating the γδ T lymphocytes. 6. The use according to any of the preceding claims, in which said at least one tumor pathology is sensitive to the action of substances with cytotoxic activity. 7. The use according to any of the preceding claims, in which said at least one tumor pathology is sensitive to the action of substances with apoptotic activity, 8. The use according to any one of the preceding claims, in which said at least one tumor pathology is selected from the group comprising: sarcoma. carcinoma, carcinoid. kidney cancer, colon cancer, breast cancer, bone cancer, neuroendocrine cancer, prostate cancer, leukemia, Bcr-Abl positive leukemia, chronic myelogenous leukemia. lymphoid leukemia, mycloid leukemia, megakaryoptic leukemia, non-Hodgkin's lymphomas. melanoma, Hodgirin's disease. tumor pathologies sensitive to immunotherapies, tumors, gastrointestinal stroma li (GlSTs). 9. Use of one or more compounds of formula (I), according to any one of claims 1 to 3. for the preparation of a pharmaceutical composition for the treatment of a pathology correlated and / or correctable to abnormal angiogenesis. 10. The use according to claim 9. wherein said pathology is selected from the group comprising: arthritis, tumors responsive to angiogenic activity. metastatic spread, diabetic retinopathy, psoriasis, chronic inflammation, atherosclerosis, The use according to any one of the preceding claims, wherein said one or more compounds of formula (1) is formulated in combination with at least one compound having antiblastic activity. The use according to claim 11, wherein said at least one compound having antiblastic activity is selected from the group comprising: alkylating agents, topoosomerase inhibitors. de) fused poison agents, intercalating agents, antimetabolites, products of natural origin, such as. Vinca alkaloids, epipodophyllotoxins. antibiotics, enzymes, taxanes, anti-cancer vaccines. 13. A compound of formula (I), according to any one of claims 1 to 3, selected from the group consisting of; 14. A compound according to claim 13 for use as a medicament. 14. A Pharmaceutical composition comprising at least one of the compounds according to claim 13. The pharmaceutical composition according to claim 14, further comprising at least one pharmacologically acceptable excipient and / or diluent.