WO2005011673A1

WO2005011673A1 - Use of antitumoral compounds

Info

Publication number: WO2005011673A1
Application number: PCT/GB2004/003177
Authority: WO
Inventors: Julia Pérez Baz; Fernando DE LA CALLE VERDÚ; Luis Francisco GARCIA FERNÁNDEZ
Original assignee: Instituto Biomar S.A.; Ruffles, Graham, Keith
Priority date: 2003-07-22
Filing date: 2004-07-21
Publication date: 2005-02-10
Also published as: GB0317128D0

Abstract

Compounds of formula (I): wherein R1 is selected from the group consisting of H, (II) and (III); or pharmaceutically acceptable salts, derivatives, prodrugs, tautomers and solvates, or stereoisomers thereof, are of use in the preparation of medicaments for the treatment of cancer.

Description

USE OF ANTITUMORAL COMPOUNDS

FIELD OF THE INVENTION

The present invention relates to the use of antitumoral macrolides for the treatment of cancer, and to pharmaceutical compositions containing them.

BACKGROUND OF THE INVENTION

Several antitumoral macrolides has been described in the literature.

For example Trends Antibiot. Res. Genet. Biosyth. Actions New Subst. 1982, 135- 145 discloses that Hitachimycin possesses antitumor activities against Ehrlich ascites carcinoma and P-388 leukaemia. No data is given. Hitachimycin has the following structure:

Another reference, Org Lett 2001, 3/ 1 : 57-59, discloses that Apoptolidin can selectively sensitise cancer cells to apoptosis with considerable potency. In the mouse B cell lymphoma cell line LYas the IC50 of this compound was 100 nM and in breast carcinoma cell line MCF-7 the ICso was 90 nM. Apoptolidin has the following structure:

Although this compound has a high cellular cytotoxicity, its deglycosylated derivative has a reduced cellular cytotoxicity.

In Org Lett 2002, 4/20: 3383-3386 compound FD-891 was shown to have in vitro cy to toxic activity against several tumor cell lines. The compound FD-891 has the following structure:

Cancer is a leading cause of death in animals and humans. Several efforts have been and are still being undertaken in order to obtain an antitumor agent active and safe to be administered to patients suffering from a cancer. The present invention provides compounds that are useful in the treatment of cancer.

SUMMARY OF THE INVENTION The present invention is directed to the use of a compound of formula I:

wherein Ri is selected from the group consisting of H,

or pharmaceutically acceptable salts, derivatives, prodrugs, tautomers and solvates, or stereoisomers thereof, in the treatment of cancer.

In a preferred embodiment the compound of formula I has the following stereochemistry:

wherein Ri is as defined above.

It is also preferred that Ri has the stereochemical formula: I -O ΛΓ ΓO OCONH₂ 'OH

OH or OH

In a more preferred embodiment the compound is Venturicidin A:

Venturicidin A was originally isolated in 1961 (Rhodes et al. Nature 192, 952-954 (1961)). It is known to have antibiotic and antifungal properties by inhibiting mitochondrial ATP synthesis (Walter et al. J. Biol. Chem., 242, 5014-8 (1967)). No mention is made in the literature about the use of Venturicidin A and related compounds as antitumoral agents.

The present invention is also directed to a pharmaceutical composition comprising a compound as previously defined and a pharmaceutically acceptable carrier, which may take the form of a vehicle or diluent.

The present invention further provides a method of treating any mammal, notably a human, affected by cancer which comprises administering to the affected individual a therapeutically effective amount of a compound as defined above. In another aspect the present invention is directed to the use of a compound as defined above in the manufacture of a medicament for the treatment of cancer.

The invention additionally provides kits comprising separate containers containing a pharmaceutical composition comprising a compound as defined above and a reconstituting agent. Methods of reconstitution are also provided.

DETAILED DESCRIPTION OF THE INVENTION

We have found that compounds of general formula I:

g of H,

have shown significant antitumoral activities, see the examples.

The term "pharmaceutically acceptable salts, derivatives, prodrugs" refers to any pharmaceutically acceptable salt, ester, solvate, hydrate or any other compound which, upon administration to the recipient is capable of providing (directly or indirectly) a compound of the invention or a metabolite or a residue thereof. However, it will be appreciated that non-pharmaceutically acceptable salts also fall within the scope of the invention since those may be useful in the preparation of pharmaceutically acceptable salts. The preparation of salts, prodrugs and derivatives can be carried out by methods known in the art. Particularly favourite derivatives and prodrugs are those that increase the bioavailability of the compounds of this invention when such compounds are administered to a patient (e.g., by allowing an orally administered compound to be more readily absorbed into the blood), enhance delivery of the parent compound to a given biological compartment, increase solubility to allow administration by injection, alter metabolism or alter rate of excretion.

The compounds of the present invention have asymmetric centers and therefore exist in different enantiomeric and diastereomeric forms. This invention relates to the use of all optical isomers and stereoisomers of the compounds of the present invention, and mixtures thereof, and to all pharmaceutical compositions and methods of treatment that may employ or contain them.

The present invention includes the compounds of the present invention, and the pharmaceutically acceptable salts thereof, wherein one or more hydrogen, carbon or other atoms are replaced by isotopes thereof. Such compounds may be useful as research and diagnostic tools in metabolism pharmacokinetic studies and in binding assays.

Salts of the compounds of the present invention may comprise acid addition salts derived from a nitrogen in the compound of formula 1 or 2. The therapeutic activity resides in the moiety derived from the compound of the invention as defined herein and the identity of the other component is of less importance although for therapeutic and prophylactic purposes it is, preferably, pharmaceutically acceptable to the patient. Pharmaceutically acceptable salts of compounds provided herein are synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts are, for example, prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or in a mixture of the two. Generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred. Examples of pharmaceutically acceptable acid addition salts include those derived from mineral acids, such as hydrochloric, hydrobromic, phosphoric, metaphosphoric, nitric and sulphuric acids, and organic acids, such as tartaric, acetic, trifluoroacetic, citric, malic, lactic, fumaric, benzoic, glycolic, gluconic, succinic and methanesulphonic and arylsulphonic, for example p-toluenesulphonic, acids.

Compounds of the invention can be synthesized by using conventional techniques of synthetic organic chemistry which are within the skill of the art. Such techniques are explained fully in literature. See, for example, Tsunashima K. et al., Tetrahedron Letters, 2001, 42, 3607-3611. Compounds of the invention can also be isolated from natural sources. For example, Venturicidin A can be isolated from Streptomyces griseolus (see, Biochem. Soc. Trans., 1974, 2/2, 202- 205).

Pharmaceutical formulations of the compounds of the invention may be adapted for administration by any appropriate route, for example by the oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) route. Such formulations may be prepared by any method known in the art of pharmacy, for example by bringing into association the active ingredient with the carrier (s) or excipient (s) .

Preferably pharmaceutical compositions of the compounds of the invention include liquid (solutions, sμspensions or emulsions) with suitable composition for intravenous administration, and they may contain the pure compound or in combination with any carrier or other pharmacologically active compounds.

The invention also provides kits comprising a pharmaceutical composition comprising a compound of the invention, or a pharmaceutically aceptable salt, derivative, prodrug or stereoisomer thereof, and a reconstituting agent. The reconstituting agent is intended for addition to the pharmaceutical composition to give a dosage form suited for administration to a patient.

Administration of the compounds or compositions of the present invention can be by intravenous infusion. Infusion times of up to 72 hours can be used, more preferably 1 to 24 hours, with either about 1 or about 3 hours most preferred. Short infusion times which allow treatment to be carried out without an overnight stay in hospital are especially desirable. However, infusion may be around 24 hours or even longer if required.

In the preferred application method, the administration is performed in cycles. An intravenous infusion of a compound of the invention is given to the patients the first week of each cycle, the patients are allowed to recover for the remainder of the cycle. The preferred duration of each cycle is of either 1, 3 or 4 weeks; multiple cycles can be given as needed. In an alternative dosing protocol, the compound of the invention is administered for say about 1 hour for 5 consecutive days every 3 weeks. Other protocols can be devised as variations.

Dose delays and/ or dose reductions and schedule adjustments are performed as needed depending on individual patient tolerance of treatments.

Although guidance for the dosage is given above, the correct dosage of the compound may change according to the particular formulation, the mode of application, and the particular situs, host and tumour being treated. Other factors like age, body weight, sex, diet, time of administration, rate of excretion, condition of the host, drug combinations, reaction sensitivities and severity of the disease shall be taken into account. Administration can be carried out continuously or periodically within the maximum tolerated dose.

The compounds and compositions of this invention may be used with other drugs to provide a combination therapy. The other drugs may form part of the same composition, or be provided as a separate composition for administration at the same time or at different time.

Antitumoral activities of these compounds include leukaemias, lung cancer, colon cancer, kidney cancer, prostate cancer, ovarian cancer, breast cancer, pancreas cancer, cervix cancer, sarcomas and melanomas.

EXAMPLE 1: ISOLATION OF VENTURICIDN A

Venturicidin A was isolated from a Streptomyces olivaceus, isolated from the branchia of the mollusc Venus casina collected at Castropol (Asturias, Spain). EXAMPLE 2; BIOASSAYS FOR ANTITUMOR SCREENING

The finality of these assays is to interrupt the growth of an "in vitro" tumor cell culture by means a continued exposure of the cells to the sample to be testing.

CELL LINES

Name N° ATCC Species Tissue Characteristics

K-562 CCL-243 human leukemia erythroleukemia (pleural effusion)

A549 CCL-185 human lung lung carcinoma "NSCL"

SK-MEL-28 HTB-72 human melanoma malignant melanoma

HT29 HTB-38 human colon colon adenocarcinoma

LoVo CCL-229 human colon colon adenocarcinoma

LoVo-Dox human colon colon adenocarcinoma (MDR)

DU-145 HTB-81 human prostate prostate carcinoma, not androgen receptors

LN-caP CRL-1740 human prostate prostate adenocarcinoma, with androgen receptors

Name N° ATCC Species Tissue Characteristics

SK-BR3 HTB-30 human breast breast adenocarcinoma, Her2/neu+, (pleural effusion)

IGROV human ovary ovary adenocarcinoma

IGROV-ET human ovary ovary adenocarcinoma, characterized as ET-743 resistant cells

HeLa CCL-2 human cervix cervix epitheloid carcinoma

HeLa-APL CCL-3 human cervix cervix epitheloid carcinoma, characterized as aplidine resistant cells

PANC1 CRL-1469 human pancreas pancreatic epitheloid carcinoma INHIBITION OF CELLS GROWTH BY COLORIMETRIC ASSAY.

A colorimetric type of assay, using sulforhodamine B (SRB) reaction has been adapted for a quantitative measurement of cell growth and viability [following the technique described by Philip Skehan, et al. (1990), New colorimetric cytotoxicity assay for anticancer drug screening, J. Natl. Cancer Inst. , 82: 1107-1112].

This form of assay employs 96 well cell culture microplates of 9 mm diameter (Faircloth, 1988; Mosmann, 1983). Most of the cell lines are obtained from American Type Culture Collection (ATCC) derived from different human cancer types.

Cells are maintained in RPMI 1640 10% FBS, supplemented with 0.1 g/1 penicillin and 0.1 g/1 streptomycin sulfate and then incubated at 37°C, 5% CO2 and 98% humidity. For the experiments, cells were harvested from subconfluent cultures using trypsin and resuspended in fresh medium before plating.

Cells are seeded in 96 well microtiter plates, at 5 x 10³ cells per well in aliquots of 195 μl medium, and they are allowed to attach to the plate surface by growing in drug free medium for 18 hours. Afterward, samples are added in aliquots of 5 μl in a ranging from 10 to 10^~8 μg/ml, dissolved in DMSO/EtOH/PBS (0.5:0.5:99). After 48 hours exposure, the antitumor effect are measured by the SRB methodology: cells are fixed by adding 50 μl of cold 50% (wt/vol) trichloroacetic acid (TCA) and incubated for 60 minutes at 4°C. Plates are washed with deionised water and dried. One hundred μl of SRB solution (0.4% wt/vol in 1% acetic acid) is added to each microtiter well and incubated for 10 minutes at room temperature. Unbound SRB is removed by washing with 1% acetic acid. Plates are air dried and bound stain is solubilized with Tris buffer. Optical densities are read on a automated spectrophotometric plate reader at a single wavelength of 490 nm.

The values for mean +/- SD of data from triplicate wells are calculated. Some parameters for cellular responses can be calculated: GI = growth inhibition, TGI = total growth inhibition (cytostatic effect) and LC = cell killing (cyto toxic effect).

Table 1 illustrates data on the biological activity of the compounds of the present invention.

Table 1. Activity Data (Molar)

Claims

Use of a compound of formula I:

wherein Ri is selected from the group consisting of H,

or pharmaceutically acceptable salts, derivatives, prodrugs, tautomers and solvates, or stereoisomers thereof, in the preparation of a medicament for the treatment of cancer.

2. The use according to claim 1, wherein the compound of formula I has the following stereochemistry

and Ri is as defined in claim 1.

3. The use according to claim 1, wherein the compound of formula I has the following formula

4. A pharmaceutical composition comprising a compound of formula I as defined in claim 1, 2 or 3, or a pharmaceutically aceptable salt, derivative, prodrug or stereoisomer thereof, and a pharmaceutically acceptable carrier.

5. A method of treating a patient affected by cancer which comprises administering to the affected individual a therapeutically effective amount of a compound of formula I as defined in claim 1, 2, or 3, or a pharmaceutically aceptable salt, derivative, prodrug or stereoisomer thereof.

6. A kit comprising separate containers containing a pharmaceutical composition comprising a compound of formula I as defined in claim 1, 2 or 3, or a pharmaceutically aceptable salt, derivative, prodrug or stereoisomer thereof, and a reconstituting agent.