AU4628299A

AU4628299A - Therapeutic composition based on flavonoids for use in the treatment of tumours with cytotoxic agents

Info

Publication number: AU4628299A
Application number: AU46282/99A
Authority: AU
Inventors: Francis Darro; Armand Frydman; Robert Kiss
Original assignee: Laboratoire L Lafon SA
Current assignee: Cephalon France SAS
Priority date: 1998-07-15
Filing date: 1999-07-13
Publication date: 2000-02-07
Anticipated expiration: 2019-07-13
Also published as: JP2002520357A; FR2781154B1; BR9912817A; CN1312712A; EP1096929A1; CN1139383C; KR20020006510A; IL140580A0; FR2781154A1; WO2000003707A1; CA2337256A1; EA200100141A1; AU761417B2

Description

WO 00/03707 PCT/FR99/01715 ISOFLAVONOID-BASED THERAPEUTIC COMPOSITION INTENDED TO BE USED IN THE TREATMENT OF TUMOURS WITH CYTOTOXIC AGENTS The present invention relates to the use of 5 compounds of the isoflavonoid type in the treatment of cancers with cytotoxic agents. A cancer is a disorder of the somatic genes in which genetic dysfunctions become amplified as the tumour process progresses from the state of a 10 precancerous lesion to that of a malignant transformation, the cancerous tumour becoming metastatic and often resistant to cytotoxic medicaments. In spite of major efforts made in all developed 15 countries, in particular through experimental and clinical research programmes, mortality due to the various cancers (solid tumours and haematological neoplasias) remains unacceptably high. In many countries, the mortality caused by cancer is ranked 20 second, just after cardiovascular diseases. In terms of newly diagnosed cancers, the distribution between solid tumours and haematological neoplasias (bone marrow, blood, lymphatic system) shows that 9 cancers out of 10 are solid tumours. Contrary to 25 what is observed in haematological oncology (therapeutic success in 40 to 90% of the cancers of the blood cells), only a small number of advanced or disseminated solid tumours respond to chemotherapy treatments alone. It is partly for this reason that the 30 overall mortality caused by cancer increased in the USA between 1973 and 1992. It is unfortunately not certain that this trend can be reversed solely by the appearance, besides the established chemotherapy arsenal, of novel antitumour 35 medicaments such as taxanes (paclitaxel and docetaxel) which interfere with the formation of the microtubules (W.P. McGuire et al., Am. Intern. Med., 1989), the inhibitors of topoisomerases I derived from camptothecin (topotecan and irinotecan) , vinorelbine - 2 (novel alkaloid derived from periwinkle), gemcitabine (novel cytotoxic antimetabolic agent), raltitrexed (inhibitor of thymidylate synthetase) and miltefosine (first representative of the alkylphosphocholine 5 family). These treatments are in addition, either as a first line treatment, or as a second line treatment, to the medicaments whose specific activity is now well recognized such as doxorubicin, cisplatin, vincristine, methotrexate, 5-fluorouracil. 10 one of the most difficult current problems of anticancer chemotherapy is due to the fact that many populations of malignant cells exhibit substantial resistance to the established cytotoxic substances. Most often, this situation results from the existence 15 of multiresistance genes or from the frequency of genetic mutations in certain types of tumours. Thus, the treatment of cancers requires novel approaches, complementary to those currently used, and intended for better combating the extension and heterogeneity of the 20 tumour load and the acquisition of "multi-cytotoxic drug" resistance. Among these novel approaches, some are already promising. That is the case for the induction of apoptosis, the inhibition of tumour angiogenesis and of 25 metastatic processes, not to mention gene therapy or immunotherapy. The inventors were interested in a different approach. The objective sought was to make the population of tumour cells more sensitive to the 30 reference anticancer treatments in order to achieve a double beneficial effect: 1) to increase the cytotoxic activity and therefore the efficacy, and 2) to reduce the frequency and the severity of 35 certain side effects by virtue of the reduction of the dosage which might follow the induction of the increase in the antitumour efficacy. It is this strategy which is at the origin of the discovery of an innovative mechanism caused by -3 substances having a low antitumour power or even lacking this power, but capable of inducing a very significant increase in the cytotoxic activity of proven anticancer medicaments. This innovative 5 mechanism results from the possibility for these substances either to stimulate the recruitment of clonogenic cells inside the tumour, making it more sensitive to conventional treatment with cytotoxic agents, or to inhibit the proliferation of clonogenic 10 cells, thus contributing to the regression of the tumour. The subject of the present invention is thus the use, in the treatment of cancers with at least one antitumour agent chosen from cytotoxic agents, of a 15 compound having an activity on the proliferation of clonogenic cells, chosen from isoflavonoids and analogous compounds of the chromone type and in particular the compounds of formula: R1

R

2 R3 R6 R4 0 20 in which formula: - R 1 , R 2 , R 3 and R 4 are chosen, independently of each other, from H, OH, a Ci-C4 alkoxy group, an -OCOR 7 25 group, R 7 being a Ci-C4 alkyl group, at least one of the substituents R 1 , R 2 , R 3 or R 4 being other than H and it being possible for R 2 and R 3 to form together a methylenedioxy group, - R 5 is chosen from H, OH, a Ci-C4 alkoxy group, an 30 0-glycosyl group and a cyclohexyl group, - R 6 is chosen from a cyclohexyl group, a phenyl group and a phenyl group substituted 1 to 3 times with groups chosen from H, OH and a Ci-C4 alkoxy group, - 4 - and - denotes either a double bond, or a single bond. A preferred class of compounds of formula I are those in which R 6 is chosen from the phenyl group, the 5 4-hydroxyphenyl group and the 4-(C 1

-C

4 alkoxy)phenyl groups. The cytotoxic agents may be used at their usual dose and, in this case, their efficacy is enhanced, or at lower doses taking into account the increase in 10 their antitumour efficacy if the desired objective is first to enhance the patient's tolerance to the treatment. The subject of the present invention is also a composition having an activity on the proliferation of 15 clonogenic cells by interfering with the generation of clonogenic cells, either by stimulating the proliferation and recruitment, or by inhibiting the proliferation, comprising a therapeutically effective quantity of an isoflavonoid or of an analogous compound 20 of the chromone type, and in particular of a compound chosen from the compounds of formula: Ri O%- R5 R2.

R

3 R6 R4 0 25 in which formula: - R 1 , R 2 , R 3 and R 4 are chosen, independently of each other, from H, OH, a Ci-C 4 alkoxy group, an -OCOR 7 group, R 7 being a Ci-C 4 alkyl group, at least one of the substituents R 1 , R 2 , R 3 or R 4 being other than H and it 30 being possible for R 2 and R 3 to form together a methylenedioxy group, -5 - R 5 is chosen from H, OH, a Ci-C 4 alkoxy group, an 0-glycosyl group, and a cyclohexyl group, - R 6 is chosen from a cyclohexyl group, a phenyl group and a phenyl group substituted 1 to 3 times with groups 5 chosen from H, OH and a Ci-C 4 alkoxy group, - and - denotes either a double bond, or a single bond. The subject of the present invention is also the use of an isoflavonoid, in particular of a compound 10 of formula I as defined above, for the manufacture of a medicament intended to interfere (by induction or inhibition) with the generation of clonogenic cells in tumours during a treatment with at least one cytotoxic agent. 15 In the chemotherapeutic treatment of cancers with cytotoxic agents, the isoflavonoids and in particular the compounds of formula I may be administered at the beginning of the chemotherapy treatments either once, or over several days at the 20 beginning of these treatments (for example for 5 to 7 days) and, depending on the chemotherapy protocol, at the beginning of each treatment cycle (for example for 2 to 5 days) during each cure. The isoflavonoids and in particular the 25 compounds of formula I are advantageously administered by infusion (generally over 1 to 3 hours) at doses of 5 to 50 mg/kg/day or 200 to 2000 mg/m 2 /day. In order to obtain a maximum effect on the production of clonogenic cells, the isoflavonoids 30 should be administered such that the tissue concentrations obtained are the highest which can be possibly envisaged. For the treatment protocols in the acute phases of the cures, the intravenous route is to be preferred 35 using: - ready-to-use infusion solutions (bags, vials and the like) intended to be administered as they are by intravenous infusion with the aid of an infusion line and using the recommended flow rate: -6 - lyophilizates to be resuspended in solution for intravenous infusion with the aid of pharmaceutical solutions known to persons skilled in the art; - for the maintenance treatments, it is also 5 possible to envisage the oral route when the chemotherapy treatment preferably uses the administration--f cytostatic agents by the oral route. For this purpose, oral lyophilizates (for oral or perlingual absorption), instant or delayed release 10 tablets, oral solutions, suspensions, granules, gelatine capsules and the like may be used. The compounds of formula (I) are, for the majority, compounds of natural origin or are derivatives of compounds of natural origin. As 15 examples, there may be mentioned: - genistein, - biochanin A, - daidzein, - formononetin, 20 - 7-acetylformononetin, - glycetein, - orobol or 5,7,3',4'-tetrahydroxyisoflavone, - irizolone or 6,7-methylenedioxy 4'-hydroxyisoflavone, 25 - irigenin or 3',5,7-trihydroxy 4',5',6-methoxyisoflavone, - tectorigenin or 4',5,7-trihydroxy 6-methoxyisoflavone, - 2-hydroxy-8-methoxy-2,3-dihydroisoflavone, 30 - 4',7-dihydroxy-5-methoxyisoflavone. Other isoflavones which can be used are described by Donnelly et al. in Natural Product Reports, 1995, 321, or can be prepared by the methods described in this article. 35 The cytotoxic agents may be chosen from: i) intercalating agents, in particular doxorubicin (adriamycin), daunorubicin, epirubicin, idarubicin, zorubicin, aclarubicin, -7 pirarubicin, acridine, mitoxanthrone, actinomycin D, eptilinium acetate; ii) alkylating agents chosen from platinum derivatives (cisplatin, carboplatin, 5 oxaliplatin and the like), iii) a compound chosen from the other groups of alkylating agents: - cyclophosphamide, ifosfamide, chlormetrin, melphalan, chlorambucil, estramustine, 10 - busulfan, mitomycin C, - nitrosoureas: BCNU (carmustine), CCNU (lomustine), fotemustine, streptozotocin, - triazines or derivatives, procarbazine, dacarbazine, 15 - pipobroman, - ethyleneimines: altretamine, triethylene thiophosphoramide, iv) a compound chosen from the other groups of antimetabolic agents: 20 - antifolic agents: methotrexate, raltitrexed, - antipyrimidines: 5-fluorouracil (5-FU), cytarabine (Ara-C), - hydroxyurea - antipurines: purinethol, thioguanine, 25 pentostatin, cladribine - inductors of the synthesis of cytotoxic nucleosides: gemcitabine, v) a compound chosen from the other groups of agents with high affinity for the tubules: 30 - vinca alkaloids which disorganize the mitotic spindle: vincristine, vinblastine, vindesine, navelbine - agents blocking the depolymerization of the mitotic spindle: paclitaxel, docetaxel 35 - agents inducing breaks in the DNA by inhibition of topoisomerase II: etoposide, teniposide - inhibitors of topoisomerase I inducing breaks in DNA: topotecan, irinotecan, -8 vi) an agent breaking, fragmenting DNA, such as bleomycin, vii) one of the following compounds: plicamycin, L asparaginase, mitoguazone, dacarbazine, 5 viii) an anticancer progestogenic steroid: medroxyprogesterone, megestrol, ix) an anticancer oestrogenic steroid: diethylstilbestrol; tetrasodium fosfestrol, x) an antioestrogen: tamoxifen, droloxifen, 10 raloxifen, aminogluthetimide, xi) a steroidal antiandrogen (e.g. cyproterone) or a nonsteroidal antiandrogen (flutamide, nilutamide)-. In particular, the compounds of formula I may 15 be combined with all the treatments with the major cytotoxic agents used in polychemotherapy of solid tumours such as: - doxorubicin - alkylating agents: oxazophorines 20 (cyclophosphamide, ifosfamide, chlorambucil, melphalan) - nitrosoureas - mitomycin C - antimetabolites such as methotrexate, 5-FU, Ara-C, capecitabine 25 - agents which interfere with tubulin: vinca alkaloids (vincristine, vinblastine, vindesine, navelbine), taxoids (paclitaxel, docetaxel), derivatives of epipodophyllotoxins (etoposide, teniposide) 30 - bleomycin - inhibitors of topoisomerase I: topotecan, irinotecan. Likewise, the compounds of formula I may be combined with the treatment with the major cytotoxic 35 agents used in oncohaematology for the treatment of blood cancers: - Hodgkin's disease: cyclophosphamide, mechlorethamine, chlorambucil, melphalan, ifosfamide, etoposide, doxorubicin, daunorubicin; -9 - acute leukaemias: methotrexate, 6-mercaptopurine, cytarabine, vinblastine, vincristine, doxorubicin, daunorubicin, L-asparaginase; - non-Hodgkin's malignant lymphomas, 5 mechlorethamine, chlorambucil, cyclophosphamide, melphalan, ifosfamide, methotrexate, cytarabine, vinblastine, vincristine, etoposide, doxorubicin, daunorubicin, carmustine, lomustine, cisplatin; - chronic lymphoid leukaemias: mechloretamine, 10 chlorambucil, cyclophosphamide, melphalan, ifosfamide. Results of pharmacological trials demonstrating the effects obtained will be given below. 1 - Interaction (stimulation or inhibition of 15 proliferation) with the generation of clonogenic cells (clonogenic test) The test used is that described by Hamburger et al. (Science, 1977; 197, 461-463) and Salmon et al. (New England J. Med., 298, 1321-1327). A cell is 20 considered to be clonogenic if it possesses the capacity to proliferate and to give rise to a cell colony. The "human tumour stem cells" are the cells which are at the origin of the neoplastic cells which constitute a given tumour. These tumour stem cells are 25 responsible for the recidivation processes which can be observed after surgical resection of the primary tumours and are also responsible for the formation of metastases. At the level of a tumour or a tumour cell line, these clonogenic stem cells are distinguishable 30 from the other cells of the tumour or the neoplastic cell line considered, by the fact that they retain their capacity to proliferate in the absence of any solid support. In this test, the tumour cells are cultured on 35 a semisolid support. Only the cells which do not require a solid support for their growth (that is to say the highly tumorigenic cells called "anchorage independent cells" by M.I. Dawson et al., Cancer Res. 1995; 55: 4446-4451; also called clonogenic cells with - 10 reference to "clonal growth") are capable of developing on such an agar-based support. Indeed, on such a medium, the normal cells - which grow in "adherent mode" ("anchorage-dependent cells" according to the 5 terminology of M.I. Dawson) - such as for example the fibroblasts, do not survive. Within a tumour cell population, cultured on such a support, it is these clonogenic cells (associated with an unlimited number of cell divisions and whose proliferation is called 10 "anchorage-independent [clonal] growth" by M.I. Dawson) which are capable of growing. The percentage of these clonogenic cells within a tumour or a cell line varies between 0.1% and 0.001%. The nonclonogenic cells (associated with a limited number of cell divisions) do 15 not develop in this test because they require a solid support for their growth which should occur in "adherent mode" ("anchorage-dependent [adherent] growth", according to M.I. Dawson et al., Cancer Res. 1995; 55: 4446-51)". 20 The influence of compounds of formula (I) on the growth of the cell colonies obtained by culturing, for example, the mammary tumour lines MCF7 and MXT and the colorectal line HT-29 on the semiliquid culture medium called "soft agar" was measured. On such a 25 medium, only the clonogenic cells called "anchorage independent (clonal) cells" by M.I. Dawson survive and develop. The growth of these cells in such a "nonadherent" mode reflects their degree of tumorigenicity. The inhibition of the growth of the 30 size of a tumour in which a larger number of clonogenic cells have developed then becomes the control for a reinforced cytotoxic activity. By contrast, this test can also reveal that a compound is capable of inhibiting the 35 generation/proliferation of clonogenic cells, which makes the tumour less capable of developing, and therefore reduces the population of tumour cells. The tumour cell lines studied are maintained in culture in 25 cm 2 falcon flasks. They are then - 11 trypsinized and the cells well dissociated from each other. The percentage of living cells is determined after staining with trypan blue. A cellular suspension at the concentration 5-104 to 15-104 cells/ml (depending 5 on the cell type considered) is prepared in a 0.3% agar solution. Next, 200 pl of this suspension are inoculated into Petri dishes 35 mm in diameter, in which 3 ml of a bottom layer consisting of a 0.5% agar solution are deposited. The 200 pil of cellular 10 suspension are in turn covered with 1.8 ml of a top layer consisting of a 0.3% agar solution. The dishes are then placed in an incubator at 370C, 5% Co 2 and 70% humidity until the treatment. The latter is performed about 1 to 2 hours after inoculation. The compounds to 15 be tested are prepared at a concentration 100-fold greater than the desired concentration and 50 pl of these treating solutions are deposited on the agar top layer of the corresponding dishes. In the present study, the final concentration of the products tested 20 is 10-s, 10- and 10~9 M. The dishes are then maintained in the incubator for 21 days. On the 21st day, the dishes are treated by depositing on the top layer 100 pl of a solution of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolinium 25 bromide) at 1 mg/ml prepared with RPMI 1640 medium for 3 h at 37 0 C. After this period of time, the cell colonies are fixed by adding 2 ml of formalin per dish. After fixing for 24 hours, the formalin is evaporated and a number of coloured cell colonies, therefore 30 consisting of metabolically active cells, and whose surface area is greater than 100 ptm 2 , is determined with the aid of an inverted microscope. The average number of clonogenic cell clones determined for each experimental condition studied is 35 expressed as a percentage relative to the average number of clonogenic cell clones counted under the control condition and posed as equal to 100%. These values, expressed as the percentage relative to the control condition, are presented in Table I.

- 12 TABLE I CELL Genistein (in mol.1 1 ) LINES 10-s 10~7 10~ MCF7 66.9 2.9 74.2 4.7 89.2 ± 0.9 ** * NS HT-29 118.2 + 2.8 108.9 2.3 104.6 ± 2.5 ** * NS MXT 71 2.5 118.5 2.2 117.5 ± 2.2 ** ** ** - The results summarized in this table represent the mean values ± standard error of the mean (SEM) 5 established on at least 6 wells. - Control condition = 100% - (NS: p>0.05; *: p<0.05; **: p<0.01; ***: p<0.001). Depending on the cell line studied, genistein 10 can: - recruit the clonogenic cells inside the tumour (cell lines HT-29 at the concentrations of 10-s M and 10-', and MXT at the concentrations of 10-7 M and 10~9 M) , that is to say induce a significant increase in the 15 number of colonies of these cells compared with that obtained under the control condition, and then makes them more sensitive to the conventional treatment with cytotoxic agents, or - be capable of directly inhibiting the proliferation 20 of these clonogenic cells (MCF7 cell line at the concentrations of 10-s M and 10-7 M) . 2 - Cytotoxic activity at the level of the nonclonogenic cells: "MTT test" 25 The influence of the compounds of formula (I) on the nonclonogenic cells was evaluated with the aid of the MTT colorimetric test. The principle of the MTT test is based on the mitochondrial reduction by metabolically active living 30 cells of the product MTT (3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide), which is yellow in - 13 colour, to a product which is blue in colour, formazan. The quantity of formazan thus obtained is directly proportional to the quantity of living cells present in the culture well(s). This quantity of formazan is 5 measured by spectrophotometry. The cell lines are maintained in monolayer culture at 37 0 C in closed-stopper culture dishes containing basal medium MEM 25 MM HEPES (Minimum Essential Medium). This medium is quite suitable for 10 the growth of a range of varied mammalian diploid or primary cells. This medium is then supplemented: - with a quantity of 5% of decomplementized SVF (Foetal Calf Serum) at 56 0 C over 1 hour, - with 0.6 mg/ml of L-glutamine, 15 - with 200 IU/ml of penicillin, - with 200 ptg/ml of streptomycin, - with 0.1 mg/ml of gentamicin. The 12 human cancer cell lines which were used were obtained from the American Type Culture Collection 20 (ATCC, Rockville, MD, USA). These 12 cell lines are: - U-373MG (ATCC code: HTB-17) and U-87MG (ATCC code: HTB-14) which are two glioblastomas, - SW1088 (ATCC code: HTB-12) which is an astrocytoma, 25 - A549 (ATCC code: CCL-185) and A-427 (ATCC code: HTB-53) which are two non-small-cell lung cancers, - HCT-15 (ATCC code: CCL-225) and LoVo (ATCC code: CCL-229) which are two colorectal 30 cancers, - T-47D (ATCC code: HTB-133) and MCF7 (ATCC code: HTB-22) which are two breast cancers, - J82 (ATCC code: HTB-1) and T24 (ATCC code: HTB-4) which are two cancers of the bladder, 35 - PC-3 (ATCC code: CRL-1435) which is a prostate cancer. From the experimental point of view, 100 pl of a cellular suspension containing 20,000 to 50,000 (according to the cell type used) cells/ml of culture - 14 medium are inoculated into flat-bottomed 96-well multi-well plates and are incubated at 37*C, under an atmosphere comprising 5% Co 2 and 70% humidity. After 24 hours of incubation, the culture medium is replaced 5 with 100 pl of fresh medium containing either the various compounds to be tested at concentrations varying from 10-5 to 10-10 M, or the solvent which served for the dissolution of the products to be tested (control condition). After 72 hours of incubation under 10 the preceding conditions, the culture medium is replaced with 100 pl of a yellowish solution of MTT dissolved in an amount of 1 mg/ml in RPMI 1640. The microplates are incubated for 3 hours at 370C and then centrifuged for 10 minutes at 400 g. The yellowish 15 solution of MTT is removed and the blue formazan crystals formed in the cell are dissolved in 100 tl of DMSO. The microplates are then placed under stirring for 5 minutes. The intensity of the resulting blue colour, and therefore of the conversion of the yellow 20 MTT product to blue formazan by the cells still alive at the end of the experiment, is quantified by spectrophotometry with the aid of a DYNATECH IMMUNOASSAY SYSTEM type apparatus at the wavelengths of 570 nm and 630 nm corresponding to the wavelengths for 25 maximum absorption of formazan and to the background noise, respectively. A software integrated into the spectrophotometer calculates the mean optical density values as well as the standard deviation (Std. Dev.) and standard error of the mean (SEM) values. 30 By way of nonlimiting example, the results of the mean optical density, expressed as a percentage relative to the mean optical density measured under the control condition (posed equal to 100%), obtained with an isoflavonoid: genistein, on the 5 tumour cell lines 35 U-87MG, J82, HCT-15, T-47D and A549, will be given in Table II.

- 15 TABLE II CELL Genistein (in mol.1~ 1 ) LINES 10-9 10-6 10-7 10-8 10-9 10-10 U-87MG 83.8 + 98.1 + 94.3 100.1 + 98.2 ± 108.6 + 3.5 4.4 3.7 6.6 3.5 2.3 ** NS NS NS NS * J82 87.0 99.3 101.6 101.8 + 102.8 104.2 + 1.0 1.1 0.8 1.8 &.5[sic] 1.5 NS NS NS NS NS HCT-15 96.8 100.9 97.5 + 89.2 89.4 ± 90.5 5.3 6.0 5.2 3.5 4.0 3.3 NS NS NS * * * T-47D 92.3 98.9 95.1 + 97.8 100.0 102.4 + 2.2 3.3 1.6 3.0 3.4 1.7 * NS NS NS NS NS A-549 81.4 + 105.0 101.6± 106.0 + 108.9 103.6 + 4.8 4.1 5.4 3.2 2.1 3.9 ** NS NS NS * NS - xx + yy = mean value standard error of the mean 5 - control condition = 100% - (NS: p >0.05; *: p <0.005; **; p <0.01; ***: p < 0.001). Genistein has a low antitumour power. This 10 nontoxic product induces, when it is the case, inhibition of the overall cell proliferation of these lines only at the concentration of 10-5 M and this inhibition does not exceed 20%. At the other concentrations tested, only a few marginal effects can 15 be demonstrated. 3. - Determination of the maximum tolerated dose (MTD): The evaluation of the maximum tolerated dose 20 was carried out in 4- to 6-week old B6D2F1/Jico mice. The compounds were administered by the intraperitoneal route in increasing doses ranging from 2.5 to - 16 160 mg/kg. The value of the MTD (expressed in mg/kg) is determined from the observation of the rate of survival of the animals over a period of 14 days after a single administration of the product considered. The variation 5 of the weight of the animals is also monitored over this period. When the MTD value is greater than 160 mg/kg, the MTD value is considered to be 160 mg/kg by default. Genistein is by default associated with an MTD 10 equal to 160 mg/kg. This result suggests that the products of the isoflavonoid family do not exhibit any direct toxicity and can be used in high tissue concentrations, and therefore in high dosages. 15 4. - Antitumour activity in vivo in combination with a cytotoxic agent The trials were carried out on the models of: - hormone-sensitive murine mammary adenocarcinoma MXT (HS-MXT), 20 - lymphoma P 388, in the presence or otherwise of cytotoxic agents such as cyclophosphamide, etoposide, doxorubicin or vincristine. When the MTD value for a product was 25 determined, its in vivo antitumour activity was characterized at the doses of MTD/2, MTD/4 and MTD/8 on the model of mammary adenocarcinoma of murine origin HS-MXT and on the lymphoma P388 model) . It is the dose which exhibited the best antitumour activity on these 30 different models which was selected and used in the context of the treatments combined with the cytotoxic agents. In all the examples presented below, whatever the model (mammary adenocarcinoma HS-MXT or lymphoma 35 P 388), the control condition is represented by a group of 9 mice to which a volume of 0.2 ml of physiological saline containing the solvent used to dissolve the different compounds of formula (I) used is administered for 5 consecutive weeks and at the rate of - 17 5 administrations (Monday, Tuesday, Wednesday, Thursday and Friday) per week. The following were determined during these trials: 5 i) - rate of survival of the mice This rate of survival was calculated in the form of a ratio T/C: (Number of days (Treated (Number of mice which of survival of median - died during the days the median mouse mouse) which preceded that of the treated for the treated median mouse group) mouse) T = -------------------------------- (Number of mice which died on the same day as the treated median mouse) (Number of days (Treated (Number of mice which of survival of median - died during the days the median mouse mouse) which preceded that of the treated for the control median mouse group) mouse) C =

+---------------------------------

(Number of mice which died on the same day as the control median mouse) This ratio represents the mean survival time for the 10 median mouse of the treated mouse group relative to the mean survival time for the median mouse of the control mouse group. Thus, a molecule induces a significant increase (P <0.05) in the survival of the animals when the T/C ratio exceeds 130%. On the other hand, it 15 exhibits a toxic effect when this T/C value is less than 70%. ii) - tumour growth by measuring, twice per week (Monday and Friday), the surface area of the transplanted HS-MXT and P388 tumours. This surface area - 18 is calculated by taking the product of the value of the two largest perpendicular axes of the tumour. The value of these axes is measured with the aid of a slide calliper. 5 4.1. Murine mammary adenocarcinoma (HS-MXT) The model of murine mammary adenocarcinoma MXT which is hormone-sensitive (HS-MXT) transplanted in 4 to 6-week old B6D2F1/Jico mice is a model derived from 10 the galactophorous ducts of the mammary gland (Watson C. et al. Cancer Res. 1977; 37: 3344-48). The results obtained using genistein either alone or in combination with the cytotoxic agents will be given by way of example. 15 Treatment 1 Genistein is administered alone. The first injection of the product is carried out on the seventh day post-transplantation (D7) for four consecutive 20 weeks at the rate of 5 injections per week (Monday, Tuesday, Wednesday, Thursday and Friday) and at the dose of 20 mg/kg. Treatment 2 Cyclophosphamide is administered alone. The 25 first injection of the product is carried out on the fourteenth day post-transplantation (D14) for three consecutive weeks at the rate of 3 injections per week (Monday, Wednesday, and Friday) and at the dose of 10 mg/kg. 30 Treatment 3 Vincristine (VCR) is administered alone. The first injection of the product is carried out on the fourteenth day post-transplantation (D14) for three consecutive weeks at the rate of 3 injections per week 35 (Monday, Wednesday, and Friday) and at the dose of 0.63 mg/kg. Treatment 4 Etoposide (ETO) is administered alone. The first injection of the product is carried out on the - 19 fourteenth day post-transplantation (D14) for three consecutive weeks at the rate of 3 injections per week (Monday, Wednesday, and Friday) and at the dose of 10 mg/kg. 5 Treatment 5 Genistein is coadministered with cyclophosphamide. In this case, the first injection of genistein is carried out on the seventh day post transplantation (D7) for four consecutive weeks at the 10 rate of 5 injections per week (Monday, Tuesday, Wednesday, Thursday and Friday) at the dose of 20 mg/kg and the first injection of cyclophosphamide is carried out on the fourteenth day post-transplantation (D14) for three consecutive weeks at the rate of three 15 injections per week (Monday, Wednesday and Friday) at the dose of 10 mg/kg. Treatment 6 Genistein is coadministered with vincristine. In this case, the first injection of genistein is 20 carried out on the seventh day post-transplantation (D7) for four consecutive weeks at the rate of 5 injections per week (Monday, Tuesday, Wednesday, Thursday and Friday) at the dose of 20 mg/kg and the first injection of vincristine is carried out on the 25 fourteenth day post-transplantation (D14) for three consecutive weeks at the rate of three injections per week (Monday, Wednesday and Friday) at the dose of 0.63 mg/kg. Treatment 7 30 Genistein is coadministered with etoposide. In this case, the first injection of genistein is carried out on the seventh day post-transplantation (D7) for four consecutive weeks at the rate of 5 injections per week (Monday, Tuesday, Wednesday, Thursday and Friday) 35 at the dose of 20 mg/kg and the first injection of etoposide is carried out on the fourteenth day post transplantation (D14) for three consecutive weeks at the rate of three injections per week (Monday, Wednesday and Friday) at the dose of 10 mg/kg.

- 20 The results obtained for the survival period (Table III) for genistein will be given below. TABLE III Treatments T/C (expressed in %) 1 (genistein) 100 2 (CPA) 107 3 (VCR) 105 4 (ETO) 116 5 (genistein + CPA) 131 6 (genistein + VCR) 135 7 (genistein + ETO) 131 5 These results show that the coadministration of genistein with the cytotoxic agents: cyclophosphamide, vincristine or etoposide, significantly increases the mean survival time for the median mouse of the 10 different groups of mice thus treated compared with the mean survival time for the median mouse of the control mouse group. Furthermore, this increase in the mean survival time for the median mouse of the different groups of mice treated with these coadministrations is 15 significantly longer than that obtained with the treatments involving genistein or these cytotoxic agents used alone. The study of tumour growth moreover showed the following results. In Table IV below are indicated, in 20 per cent, the decreases (-) or the increases (+) in the surface area of the HS-MXT tumours induced with the different treatments 1, 2, 3, 4, 5, 6 and 7 compared with the control condition on the 28th day after the tumour transplantation, that is after 25 15 administrations of genistein and 6 administrations of the different cytotoxic agents used or otherwise in coadministrations with genistein. On the 28th day post transplantation, 89% of the control animals are still alive (that is 8 animals out of 9). 30 - 21 Table IV Treatments Variation in the tumour surface area (expressed in %) 1 (genistein) + 2.6 2 (CPA) - 25 3 (VCR) - 32 4 (ETO) - 22 5 (genistein + CPA) - 20 6 (genistein + VCR) - 45 7 (genistein + ETO) - 41 These results show that the coadministration of 5 genistein with the cytotoxic agents: vincristine and etoposide, significantly induces a decrease in the growth of the HS-MXT tumours which is greater than that induced by the treatments involving genistein alone (which has no relevant clinical effect) or the latter 10 two cytotoxic agents used alone. 4.2. Lymphoma P 388: The 4- to 6-week old CDFl mice receive a transplant consisting of a piece of P388 tumour 15 (obtained from a bank of tumours maintained in the laboratory) subcutaneously on the right side on day DO. In order to be in a situation similar to the clinical reality, we wait for the 5th day post-transplantation (D5) before starting the treatment. This was because, 20 after this period of time, the subcutaneous P388 tumours are palpable. By way of example, the results obtained with genistein alone or in combination with vincristine are reported below. 25 Treatment 1 Genistein is administered alone. The first injection of the product is carried out on the fifth day post-transplantation (D5) at the rate of 5 injections per week (Monday, Tuesday, Wednesday, - 22 Thursday and Friday) for five consecutive weeks and at the dose of 40 mg/kg. Treatment 2 Vincristine (VCR) is administered alone. The 5 first injection of the product is carried out on the fifth day post-transplantation (D5) at the rate of 3 injections per week (Monday, Wednesday and Friday) for three consecutive weeks and at the dose of 0.63 mg/kg. 10 Treatment 3 Genistein is coadministered with vincristine. In this case, the first injection of genistein is carried out on the fifth day post-transplantation (D5) at the rate of 5 injections per week (Monday, Tuesday, 15 Wednesday, Thursday and Friday) for five consecutive weeks at the dose of 40 mg/kg and the first injection of vincristine is carried out on the fifth day post transplantation (D5) at the rate of 3 injections per week (Monday, Wednesday and Friday) for three 20 consecutive weeks at the dose of 0.63 mg/kg. The results obtained with treatments 1, 2 and 3 on the survival times for the mice are presented below in Table 5. Table V 25 Treatments T/C (expressed in %) 1 (genistein) 125 2 (VCR) 122 3 (genistein + VCR) 169 These results show that the coadministration of genistein with vincristine increases in a very highly significant manner the mean survival time for the 30 median mouse of the different groups of mice thus treated compared with the mean survival time for the median mouse of the control mouse group. Furthermore, this increase in the mean survival time for the median mouse of the different groups of mice thus treated is 35 highly significant compared with the mean survival time - 23 for the median mouse of the different groups of mice treated with genistein or vincristine which are used alone. Examples of the modality of using the compounds 5 of formula I in mono- or polychemotherapy protocols with cytotoxic agents will be given below. A. Solid tumours 1/ Lung cancers 10 1.1. Non-small-cell type (advanced stage): - to the recommended protocol (T. Le Chevalier et al., J. Clin. Oncol. 1994; 12. 360-367), the intravenous infusions of genistein or of another isoflavonoid are 15 added: Dose Route Days " isoflavonoid 200-2000 mg/m 2 /day Di, Ds, D 15 , or 5 - 50 mg/kg/day i.v. D 2 2 , D 2 9 and D 36 infusion of 1 h " navelbine 30 mg/m 2 /day i.v. D 1 , Da, D 15 ,

D

2 2 , D 2 9 and D 3 6 e cisplatin 120 mg/m 2 i.v. Di and D 2 9 this cure is repeated 8 times. 1.2. Small-cell type (advanced stage): 20 - to the recommended CAV or VAC protocol (B.J. Roth et al., J. Clin. Oncol. 1992; 10: 282-291), the isoflavonoid infusions are added: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D, infusion of 1 h " cyclophophamide 1000 mg/m 2 bolus i.v. Di " doxorubicin 40 to 50 mg/m 2 bolus i.v. D, e vincristine 1 to 1.4 mg/m 2 bolus i.v. Di (max 2 mg) - 24 this cure is to be repeated 6 times every 21 days. - to the recommended Pt-E protocol (B.J. Roth et al., J. Clin. Oncol. 1992; 10: 282-291) the genistein 5 infusions are added Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 infusion of 1 h * cisplatin 20 mg/m 2 /day infusion of 20 to i.v. Di-D 5 60 minutes e etoposide 80 mg/m 2 /day infusion of i.v. Di - Ds 60 minutes I each cycle is repeated every 21 days and the cure comprises 6 cycles. 10 1.3. Non-small-cell bronchial cancer, locally advanced or metastatic: e monochemotherapy: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day D 1 , D 8 , D 15 or 5 - 50 mg/kg/day i.v. then 1 infusion of 1 h week/rest a gemcitabine 1000 mg/m 2 /day

D

1 , D 8 , D 15 infusion of i.v. then 0.5 hour 1 week/rest it being possible for the cure to comprise the 15 repetition of the cycle of 4 weeks. e gemcitabine/cisplatin combination: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 , D 8

-D

15 infusion of 1 h I I _I - 25 * gemcitabine 1000 mg/m 2 /day infusion of i.v. D 1 , D 8 , D 15 0.5 hour * cisplatin 20 mg/m 2 /day i.v. Di infusion of 20-60 minutes the cure comprising the repetition of this cycle every 21 days. 2/ Breast cancers 5 - CMF protocol as adjuvant treatment for operable breast cancer (G. Bonnadonna et al., N. Engl. J. Med.; 1976; 294: 405-410): Dose Route Days * isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di to D 1 4 infusion of 1 h " cyclophosphamide 100 mg/m 2 /day oral Di to D 1 4 * methotrexate 40 mg/m 2 bolus i.v. Di and D 8 * 5-FU 600 mg/m 2 i.v. Di and D 8 each cycle is repeated every 28 days and the 10 cure comprises 6 cycles. - AC protocol (B. Fisher et al., J. Clin. Oncol.; 1990; 8: 1483 - 1496) as adjuvant treatment: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di infusion of 1 h * doxorubicin 60 mg/m 2 i.v. Di bolus " cyclophosphamide 600 mg/m 2 i.v. Di bolus 15 each cycle is repeated every 21 days and the cure comprises 4 cycles. - Breast cancers with metastases: - 26 - in the FAC protocol (A.U. Buzdar et al., Cancer 1981; 47: 2537-2542) and its different adaptations, the isoflavonoid infusions are added according to the following scheme 5 (nonlimiting): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D 5 and Ds or 5 - 50 mg/kg/day i.v. D 12 or Di-D 5 infusion of 1 h * 5-FU 500 mg/m 2 /day i.v. Di and D 8 or bolus Di-D2 * doxorubicin 50 mg/m 2 i.v. Di or Di and bolus D2 * cyclophos- 500 mg/m 2 bolus Di phamide i.v. or oral Di each cycle is repeated every 3 weeks until a new progression of the disease is diagnosed. - in the CAF protocol (G. Falkson et al., 10 Cancer 1985; 56: 219-224): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D14 infusion of 1 h * cyclophos- 100 mg/m 2 /day oral Di-D 14 phamide e doxorubicin 30 mg/m 2 i.V. Di and D 8 * 5-FU 500 mg/m 2 i.v. Di and D 8 bolus each cycle is repeated every 28 days until a new progression of the disease is diagnosed. - in the CMF protocol: - 27 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 and infusion of 1 h

D

8

-D

12 e cyclophos- 600 mg/m 2 /day i.v. Di and D 8 phamide bolus e methotrexate 40 mg/m 2 /day i.v. Di and D 8 bolus e 5-FU 600 mg/m 2 /day i.v. Di and D 8 bolus this cycle is to be repeated every 3 to 5 weeks and the cure comprises 6 cycles. - in the CMF-VP protocol: 5 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D 5 or 5 - 50 mg/kg/day i.v. D 8

-D

12 infusion of 1 h D 15 -Dig D22-D26 " cyclophos- 2 to 2.5 mg/kg/day oral daily phamide " methotrexate 25 to 50 mg/m 2 /day i.v. D 1 , D 8 , D 15 , D 22 " 5-FU 300 to 500 mg/m 2 /day i.v. Di, D 8 , D 15 , D 22 " vincristine 0.6 to 1.2 mg/m 2 /day i.v. Di, D 8 , D 15 , D 22 e prednisone 30 mg/m 2 /day oral from Di to Din this cure is to be repeated every 4 weeks. - in the FEC protocol: Dose Route Days " isoflavonoid 200-2000 mg/m 2 /day Di-Ds or 5 - 50 mg/kg/day i.v. and D 8

-D

12 infusion of 1 h " 5-FU 600 mg/m 2 /day i.v. Di and D 8 " epirubicin 50 mg/m 2 i.v. Di e cyclophos- 600 mg/m 2 i.v. Di phamide II _I this cure is to be repeated every 3 weeks.

- 28 - in the MMC-VBC protocol (C. Brambilla et al., Tumori, 1989; 75: 141-144): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D, or 5 - 50 mg/kg/day i.v. and D 15

-D

19 infusion of 1 h * mitomycin C 10 mg/m 2 i.v. D1 bolus * vinblastine 50 mg/m 2 /day i.v. Di and D 15 bolus this cure is to be repeated every 28 days until 5 progression of the disease is diagnosed. - in the NFL protocol (S.E. Jones et al., J. Clin. Oncol. 1991; 9: 1736 - 1739): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D, infusion of 1 h * mitoxantrone 10 mg/m 2 i.v. Di bolus * 5-FU 1000 mg/m 2 as an infusion of i.v. Di-D 3 24 hours e leucovorin 100 mg/m 2 i.v. D 1 bolus the cure comprises two cycles 21 days apart and 10 then requires evaluation. The isoflavonoid infusions may also be combined with the treatment of breast cancers with metastases when a taxoid is used, for example: - with paclitaxel (F.A. Holmes et al., J. Natl 15 Cancer Inst. 1991; 83: 1797 - 1805) in the treatment of the forms with metastases which may be resistant to anthracyclines: - 29 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 infusion of 1 h e paclitaxel 175 mg/m 2 as an i.v. D, infusion of 3 to 24 hours This cycle is repeated every 21 days until a new progression of the disease is diagnosed. - with docetaxel (C.A. Hudis et al., J. Clin. 5 Oncol. 1996; 14: 58-65), in locally advanced or metastatic breast cancer, resistant or in relapse after cytotoxic chemotherapy (which comprised an anthracycline) or in relapse during an adjuvant treatment: 10 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-Ds infusion of 1 h * docetaxel 100 mg/m 2 or i.v. D, 60-100 mg/m 2 as an infusion of 1 hour (or of 24 hours) This cycle is repeated every 21 days for a cure of 2 cycles or until a progression of the disease appears. - in the dose intensification protocols 15 combining a transplantation of autologous medullary cells and peripheral blood stem cells as a consolidation of the first line treatment, for example: - CPB protocol (W.P. Peters et al., J. Clin. 20 Oncol. 1993; 11: 132 - 1143), in which the i.v. infusion of stem cells takes place on days D- 1 , Do and D 1

:

- 30 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D-6 to D_ 1 infusion of 1 h " cyclophosphamide 1875 mg/m2 as an i.v. D- 6 to D_ 4 infusion of 1 hour e cisplatin 55 mg/m 2 /day i.v. D- 6 to D_ 4 as a continuous infusion of 24 hours * carmustine 600 mg/m 2 /day as an i.v. D_3 (BCNU) infusion of 2 hours - CTCb protocol (K. Antman et al., J. Clin. Oncol. 1992; 10: 102-110), in which the i.v. infusion of stem cells takes place on day DO: 5 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D.

7 to D_ 1 infusion of 1 h " cyclophosphamide 1500 mg/m2 as a continuous infusion i.v. D_7 to D_ 3 of 24 hours (4 doses) e thiotepa 125 mg/m 2 as a continuous i.v. D- 7 to D_ 3 infusion of 24 hours (4 doses) e carboplatin 200 mg/m 2 as a continuous i.v. D-, to D-3 infusion of 24 hours (4 doses) - CTM protocol (L.E. Damon et al., J. Clin. Oncol. 1989; 7: 560-571 and I.C. Henderson et al., J. Cellular Biochem. 1994 (Suppl 18B): 95) 10 in which the i.v. infusion of haematopoietic stem cells takes place on Do: - 31 Dose Route Days isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D-6 to D-i infusion of 1 h * cyclophosphamide 1500 mg/m 2 /day as an i.v. D- 6 to D-3 infusion of 1 hour * thiotepa 150 mg/m 2 /day as an infusion of i.v. D- 6 to D_ 3 2 hours * mitoxantrone 10 - 15 mg/m 2 as an i.v. D- 6 to D-3 infusion of 1 hour 3/ Gynaecological cancers 5 3.1. Ovarian cancer: - for the treatment of in particular metastatic ovarian carcinomas: i) PAC protocol (G.A. Omura et al. J. Clin. Oncol. 1989; 7: 457 - 465) : the infusions of 10 isoflavonoids are administered according to the following scheme: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-Ds infusion of 1 h * cisplatin 50 mg/m 2 (or 40-90 mg/m 2 ) i.v. Di infusion of 1 to 2 hours e doxorubicin 50 mg/m 2 bolus (or 30 to 50 mg/m 2 ) i.v. Di e cyclophosphamide 1000 mg/m 2 infusion of 1 to 2 hours i.v. Di (or 200 to 600 mg/m 2 ) this cycle is repeated every 21 to 28 days and the cure comprises 8 cycles.

- 32 ii) altretamine protocol, according to A. Marietta et al. (Gynecol. Oncol. 1990; 36: 93-96): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-Ds or 5 - 50 mg/kg/day i.v. D 8

-D

12 infusion of 1 h * altretamine 200 mg/m 2 /day divided into 4 oral Di-D 15 doses 5 the cure comprising two cycles, 28 days apart. ii) paclitaxel protocol: the isoflavonoids may be added to the paclitaxel protocol as has been described by W.P. McGuire et al. (Ann. Intern. Med. 1989; 111: 273-279): 10 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D 3 or 5 - 50 mg/kg/day i.v. infusion of 1 h * paclitaxel 135 mg/m 2 infusion of 3 hours i.v. Di or of 24 hours the cure comprising two of these cycles, 28 days apart (with evaluation at the end). - for the treatment of metastatic and refractory ovarian carcinomas, the isoflavonoids may be 15 added to the second line protocol, based on topotecan: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D, infusion of 1 h " topotecan 1.5 mg/m 2 /day infusion of 0.5 i.v. Di-Ds hour - 33 the cure comprising two cycles, 21 days apart (with evaluation at the end) according to A.P. Kudelka et al. (J. Clin. Oncol. 1996: 14: 1552-1557). 5 3.2 Trophoblastic tumours: - in low-risk patients, the isoflavonoids may be combined with the protocol described by H. Takamizawa et al. (Semin. Surg. Oncol. 1987; 10 3: 36 - 44): Dose Route Days " isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-Ds infusion of 1 h * methotrexate 20 mg/day i.m. D 1 -D, (MTX) e dactinomycin 0.5 mg/day as a i.v. D 1 -Ds (DACT) bolus (MTX-DATC protocol). 3.3 Uterine cancers: 15 - the isoflavonoids may also be combined with the CAV (or VAC) protocol according to the scheme below: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 3 infusion of 1 h e cyclophosphamide 750 - 1200 mg/m 2 i.v. Di as an infusion * doxorubicin 45-50 mg/m 2 i.v. D1 as an infusion * vincristine 1.4 mg/m 2 i.v. D 1 the cure comprising a repetition of this cycle 20 every 21 days. - in the FAP protocol: - 34 Dose Route Days * isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-Ds infusion of 1 h " fluorouracil 600 mg/m 2 /day i.v. D 1 , D 8 (5-FU) e doxorubicin 30 mg/m 3 i.v. D * cisplatin 75 mg/m 2 i.v. Di the cure comprising the repetition of this cycle every 21 or 28 days. 4/ Testicular and prostate cancers 5 - the isoflavonoids may also be combined with the testicular cancer protocols: BEP protocol: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D5 infusion of 1 h * bleomycin 30 mg/m 2 i.v. Di as an infusion e etoposide 100 mg/m 2 /day i.v. Di-Ds as an infusion * cisplatin 20 mg/m 2 /day i.v. Di-Ds the cure comprising three cycles, at the rate of one cycle every 21 days. 10 5/ Bladder cancers - the isoflavonoids may be combined with the CISCA2 (also called PAC) protocol Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D, infusion of 1 h e cisplatin 50 mg/m 2 i.v. Di e cyclophosphamide 600 mg/m 3 i.v. Di as an infusion e doxorubicin 75 mg/m 2 i.v. Di - 35 as an infusion the cycle having to be repeated every 3 weeks. - in the MVAC protocol (according to CN Sternberg et al., J. Urol. 1988; 139: 461-469): Dose Route Days * isoflavonoid 200-2000 mg/m 2 /day Di-D3 or 5 - 50 mg/kg/day i.v. D 15 -Dis infusion of 1 h D22-D2s " methotrexate 30 mg/m 2 bolus i.v. Di, D 15 , D22 * vinblastine 3 mg/m 2 i.v. D 2 or D 2 ,

D

15 , D 2 2 * doxorubicin 30 mg/m 2 bolus i.v. D 2 * cisplatin 70-100 mg/m 2 i.v. Di or D 2 infusion of 1 h 5 this cycle being repeated every 4 to 5 weeks, at least for 2 cycles. 6/ Nasopharyngeal carcinomas/head and neck cancers - The isoflavonoids may be legitimately 10 combined with the polychemotherapy protocols used in the treatment of these cancers: 6.1 Nasopharyngeal cancers: - ABVD protocol: 15 Dose Route Days * isoflavonoid 200-2000 mg/m 2 /day Di-D 3 or 5 - 50 mg/kg/day i.v. DB-Dio infusion of 1 h or D 15

-D

17 " doxorubicin 30 mg/m 2 /day i.v. Di and D 8 or D 15 * bleomycin 10 mg/m 2 /day i.v. Di and D 8 or D 15 * vinblastine 6 mg/m 2 /day i.v. Di and D 8 or D 1 . * dacarbazine 200 mg/m 2 /day i.v. Di and D 8 or

D

15 - 36 the cure comprising 1 to 6 cycles repeated at the rate of 1 cycle every 4 weeks. 6.2 Head and neck cancers with metastases: 5 - in the Pt-FU protocol (e.g.: for cancers of the pharynx) : according to the DVAL Study Group (New Engl. J.M. 1991; 324: 1685 - 1690): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 infusion of 1 h * cisplatin 100 mg/m 2 i.v. Di infusion of 1 h * fluorouracil 1000 mg/m 2 /day i.v. Di-Ds (5-FU) continuous infusion the cure comprising two cycles, at the rate of 10 1 cycle every 3 weeks. 7/ Carcinomas of the soft tissues - The isoflavonoids may be introduced in a protocol such as the CYVADIC protocol: 15 - according to H.M. Pinedo et al. (Cancer 1984; 53: 1825): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D 3 or 5 - 50 mg/kg/day i.v. Ds-Dio infusion of 1 h D 15

-D

17 " cyclophosphamide 500 mg/m 2 bolus i.v. D 2 (Cy) * vincristine (V) 1.5 mg/m 2 /day bolus i.v. D 1 , D 8 ,

D

15 e doxorubicin (A) 50 mg/m 2 bolus i.v. D2 e dacarbazine 250 mg/m 2 /day i.v. Di-Ds (DIC) infusion of 15 minutes - 37 the cure comprising the repetition of this cycle every 4 weeks, first for 2 cycles. 8/ Hormone-refractory prostate cancer, with 5 metastases - in the VBL-estramustine, according to G.R. Hudis et al. (J. Clin. Oncol. 1992; 10: 1754:1761): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D3, or 5 - 50 mg/kg/day i.v. D 8 -Dio infusion of 1 h D 15

-D

17 .

D

2 2

-D

24 , D29 -D31,I

D

36 -D38 " vinblastine 4 mg/m 2 /day bolus i.v. D 1

,'D

8

,D

15 ,

D

2 2 , D 2 9 ,

D

36 e estramustine 200 mg/m 2 /day tid oral every day (600 mg/m 2 /day) for 6 weeks 10 a treatment cycle lasting for 6 weeks and being followed by 2 weeks of free interval. 9/ Cancers of the germ cells i) for tumours with a favourable prognosis: 15 - Pt-E protocol, according to G.J. Bosl et al. (J. Clin. Oncol. 1988: 6: 1231-1238) Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D 1 -Ds infusion of 1 h * cisplatin 20 mg/m 2 /day (Pt) infusion of 20 to i.v. Di-Ds 60 minutes e etoposide 100 mg/m 2 /day (E) infusion of 1 hour i.v. Di-Ds - 38 the cure comprising 4 cycles, at the rate of 1 cycle every 21 or 28 days. ii) for tumours with metastases: - PEB protocol, according to S.D. Williams et al. 5 (N. Eng. J. Med. 1987; 316: 1435-1440): Dose Route Days e isoflavonoi7 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 infusion of 1 h D 9 -Duj

D

16

-D

18 * cisplatin 20 mg/m 2 /day (P) infusion of 20 to i.v. Di-Ds 1 h e etoposide 100 mg/m 2 /day (E) infusion of 1 h i.v. D 2 , D 9 , D 16 " bleomycin 30U (or mg)/day i.v. Di-D 5 (B) bolus I _ II the cure comprising 4 cycles, at the rate of 1 cycle every 21 days. 10/ Kidney cancers 10 - metastatic renal carcinoma: the isoflavonoids may be introduced in the protocol described by M.J. Wilkinson et al. (Cancer 1993; 71: 3601-3604): 15 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-Ds infusion of 1 h D 8 -Di 5 * floxuridine 0.075 mg/kg/day i.v. Di-D 14 continuous infusion the cure comprising two cycles 28 days apart. - nephroblastoma: the isoflavonoids may be introduced in the DAVE protocol: - 39 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D1-D, infusion of 1 h D 8 -Dio e dactinomycin 0.6 mg/m 2 /day i.v. D 1 , D, " doxorubicin 30 mg/m 2 /day i.v. D 1 , D, e cyclophosphamide 200 mg/m 2 /day i.v. D 1 , D 8 infusion of 1 hour at the rate of one cycle every 3 to 4 weeks. 11/ Cancers of the digestive tube 5 11.1 Cancers of the oesophagus: - the isoflavonoids may be introduced in the FAP protocol according to: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D3 infusion of 1 h D 8 -Dio e 5-fluorouracil 600 mg/m 2 i.v. D 1 , D 8 (5-FU) e doxorubicin 30 mg/m 2 i.v. Di , cisplatin 75 mg/m 2 i.v. Di this cycle being repeated every 3 to 4 weeks. 10 11.2 Stomach cancers - in advanced gastric carcinomas and/or with metastases: - EAP protocol (according to P. Preusser et al., 15 J. Clin. Oncol. 1989; 7: 1310): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-Ds, infusion of 1 h D8-Dio e etoposide 120 mg/m 2 /day i.v. D 3 , D 4 , D 5 infusion of 1 hour or D 4

-D

6 * doxorubicin 20 mg/m 2 /day bolus I i.v.

D

1 , D 7 - 40 " cisplatin 40 mg/m 2 /day i.v. D 2

,D

8 infusion of 1 hour at the rate of 1 cycle every 28 days. - FAMtx protocol: according to J.A. Wils et al. (J. Clin. Oncol. 1991; 89; 827): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D 1

-D

3 infusion of 1 h * fluorouracil 1500 mg/m 2 bolus i.v. D, (5-FU) (F) 1 hour methotrexate " doxorubicin (A) 30 mg/m 2 bolus i.v. Dis e methotrexate 1500 mg/m 2 infusion i.v. D, (Mtx) of 30 minutes 5 the cure first comprising two cycles, 28 days apart. - in certain patients, the protocol or its variant (epirubicin replacing doxorubicin) may be used according to the following scheme: 10 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D, infusion of 1 h e fluorouracil 1500 mg/m 2 i.v. Di (5-FU) e doxorubicin (A) 30 mg/m 2 bolus i.v. Di = FAMTx or 60 mg/m 2 bolus e epirubicin (A) i.v. Di = FEMTx e methotrexate 1500 mg/m 2 i.v. Di (to be infused before 5-FU) e leucovorin 15 mg/m 2 /day oral D 2

-D

4 12/ Colorectal cancers - the isoflavonoids may be introduced in the protocol for FU-Levamizole adjuvant treatment - 41 of colorectal cancer (according to C.G. Moertel et al., N. Eng. J. Med. 1990; 322: 352): Dose Route Days " isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D1-Ds infusion of 1 h D 2 9

-D

3 1 * 5-fluorouracil 450 mg/m 2 /day bolus i.v. Di-D, e 5-fluorouracil 450 mg/m 2 bolus i.v. D 2 9 e levamisole 50 mg tid oral 3 days/week one week out of two the treatment in the form of a bolus with 5-FU 5 being repeated every week after the Dj-D 5 induction phase, for 52 weeks; that with an isoflavonoid being repeated at the same rate, the day of the 5-FU bolus and then the next 2 days. 10 - for the treatment of colorectal cancer which is refractory to treatment with 5-fluorouracil (5-FU) and with metastases: - according to M.L. Rothenberg et al. (J. Clin. Oncol. 1996; 14: 1128-1135): 15 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D1-D3, infusion of 1 h D8-Dio, Di5-Di7,

D

2 2

-D

2 4 " irinotectan 125 mg/m 2 /day i.v. D 1 , D 8 , D 15 , D22 the cure comprising two cycles, 42 days apart. 13/ Kaposi's sarcomas - the isoflavonoids may be combined with the two 20 protocols using antracyclines formulated in the form of liposomes: - 42 i) protocol described by P.S. Gill et al. (J. Clin. Oncol. 1995; 13: 996-1003) and C.A. Presant et al. (Lancet 1993; 341: 1242-1243): 5 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D3 infusion of 1 h and D 1 5

-D

1 7 e liposomal 20 mg/m 2 /day i.v. D 1 , D 15 daunorubicin infusion of 1 hour the cure comprising two cycles repeated at an interval of 28 days before evaluating the effects. ii) protocol of M. Harrison et al. (J. Clin. Oncol. 1995; 13: 914-920): 10 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D 1

-D

3 infusion of 1 h * liposomal 20 mg/m 2 i.v. Di doxorubicin infusion of 30 minutes the cure comprising two cycles repeated at an interval of 28 days before evaluating the effects. 14/ Metastatic melanomas 15 - The isoflavonoids may also be incorporated into the combined protocols for treating metastatic malignant melanomas: - DTIC/TAM protocol: according to G. Cocconi et al. (N. Eng. J. Med. 1992; 327: 516), the cure 20 comprising the repetition of 4 cycles, at the rate of 1 cycle every 21 days, according to the following scheme: - 43 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 infusion of 1 h * dacarbazine 250 mg/m 2 /day i.v. Di-D5 (DTIC) infusion [15 to 30 min if central catheter] or [30 min if peripheral infusion in 250 ml] * tamoxifen (TAM) 20 mg/m 2 /day oral I _Di-Ds the cure comprising 4 cycles at the rate of 1 cycle every 21 days. 5 15/ Neuroendocrine carcinoma - the isoflavonoids may be combined with the protocol described by C.G. Moertel et al. (Cancer 1991; 68: 227): - Pt-E protocol: 10 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. DI-D 3 infusion of 1 h * etoposide 130 mg/m 2 /day i.v. Di-D 3 infusion of 1 hour e cisplatin 45 mg/m 2 /day i.v. D 2 , D 3 infusion of 1 hour the cure comprising two cycles repeated every 28 days. 16/ Pancreatic cancer 15 - advanced-stage pancreatic adenocarcinoma: the isoflavonoids may be combined with the treatment with gemcitabine according to the protocol of M. Moore et al. (Proc. Am. Soc. Clin. Oncol. 1995; 14: 473): - 44 Dose Route Days " isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 3 , D 8 10 , infusion of 1 h D 15 , D 22 ,

D

2 9 , D 36 , D43, D57 " gemcitabine 1000 mg/m 2 i.v. D 1 , D 8 , D 15 , infusion of 0.5 D 22 , D 29 , hour

D

36 , D 43 , then

D

57 then once/week for 3 weeks then 1 week rest and evaluation B. Oncohaematology 5 1/Acute adult leukaemias 1.1. Acute lymphoblastic leukemia: 1.1.1. Linker protocol The isoflavonoids may be added to the Linker 10 protocols - induction chemotherapy and consolidation chemotherapy (see C.A. Linker et al. Blood 1987; 69: 1242-1248 and C.A. Linker et al. Blood 1991; 78: 2814-2822) according to the following schemes: i) induction chemotherapy: 15 Dose Route Days * isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 , infusion of 1 h D 8

-D

12 ,

D

15 -Dig - 45 " daunorubicin 50 mg/m 2 bolus every i.v. D 1 , D 2 , D 3 24 hours (30 mg/m2 in patients of over 50 years) * vincristine 2 mg bolus i.v. Di, D 8 , D 15 , D22 * prednisone 60 mg/m 2 /day oral D1-D28 * L-asparaginase 6000 U/m 2 i.m. D 1 7

-D

2 3 ii) consolidation chemotherapy (regime A): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. DI-D 5 , D 8

-D

12 infusion of 1 h e daunorubicin 50 mg/m 2 bolus every i.v. Di, D 2 24 hours e vincristine 2 mg bolus i.v. D 1 , D 8 , e prednisone 60 mg/m 2 /day divided oral Di-D14 into 3 doses e L-asparaginase 12,000 U/m 2 i.m. D 2 , D 4 , D 7 , D, and D 14 the consolidation cure A comprises 5 4 consecutive cycles as that described above = cycles 1, 3, 5 and 7. iii) consolidation chemotherapy (regimes B and C): The regimes described below correspond to the 10 consolidation cycles 2, 4, 6 and 8 (regime B) and 9 (regime C), described by C.A. Linker et al.: regime B: Dose Route Days * isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 , D 8

-D

12 infusion of 1 h * Ara-C 300 mg/m 2 infusion i.v. D 1 , D 4 , D 8 , of 2 hours Du - 46 e teniposide 165 mg/m 2 infusion i.v. D 1 , D 4 , D 8 , of 2 hours D 11 (4 cycles) regime C: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D, infusion of 1 h e methotrexate 690 mg/m 2 continuous i.v. Di-D2 infusion of 42 hours e leucovorin 15 mg/m 2 every oral D 2

-D

5 6 hours 1.1.2. Hoelzer protocol 5 The claimed products may be added to the cytotoxic agents of this polychemotherapy protocol (D. Hoelzer et al., Blood 1984; 64: 38-47, D. Hoelzer et al., Blood 1988; 71: 123-131) according to the following scheme: i) induction chemotherapy/Phase 1: 10 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 , D 8

-D

12 infusion of 1 h D 15 -Dig * daunorubicin 25 mg/m 2 i.v. D 1 , D 8 , D 1 ,

D

2 2 * vincristine 1.5 mg/m 2 (maximum i.v. D 1 , D 8 , D 15 , 2 mg) D22 * prednisone 60 mg/m 2 oral D 1

-D

28 * L-asparaginase 5000 U/m 2 i.m. D1-D14 (maximum 2 mg) ii) induction chemotherapy/phase 2: The phase 2 of the induction may be carried out as follows: - 47 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D 2 9

-D

33 , D 36

-D

40 ,' infusion of 1 h D43-D47 e cyclo- 650 mg/m 2 i.v. D 29 , D 4 3 , D 57 phosphamide (maximum 1000 mg) e cytarabine 75 mg/m 2 /day i.v. D 3 1

-D

34

,D

3 s-D 4 1 , infusion of 1 h D 4 5

-D

48 , D 52

-D

55 * mercapto- 60 mg/m 2 oral D 2 9

-D

5 7 purine * methotrexate 10 mg/m 2 /day i.v. D 31 , D 38 , D 45 , Ds 2 (maximum 15 mg) iii) reinduction chemotherapy/phase 1: Dose Route Days * isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 , D 8

-D

12 , infusion of 1 h D 15 -Dig, D 2 2

-D

2 6 e doxorubicin 25 mg/m 2 /day i.v. D 1 , D 8 , D 1 , D 2 2 * dexamethasone 10 mg/m 2 /day oral Di-D 28 e vincristine 1.5 mg/m 2 /day oral D 1 , D 8 , D 15 and (maximum 2 mg) D 2 2 5 iv) reinduction chemotherapy/phase 2: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D 3 1

-D

35 , D 3 8

-D

4 2 infusion of 1 h * cyclophos- 650 mg/m 2 i.v. D 2 9 phamide (maximum: 1000 mg) * cytarabine 75 mg/m 2 i.v. D 3 1

-D

3 4 , D 38

-D

4 1 * thioguanine 60 mg/m 2 oral D29-D42 - 48 1.2. Acute myeloid leukaemias: 1.2.1. Treatment of adults of any age The isoflavonoids may be added, according to 5 the scheme below, to the treatme t incorporating the standard dose of cytarabine previously described by R.O. Dilleman et al. (Blood, 1991; 78: 2520-2526), Z.A. Arlin et al. (Leukemia 199); 4: 177-183) and P.H. Wiernik et al. (Blood 1992; 79: 313-319): 10 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-Di2 infusion of 1 h * cytarabine 100-200 mg/m 2 /day i.v. Di-D, as a continuous infusion e daunorubicin 45 mg/m 2 /day as a i.v. Di-D 3 , or bolus

D

8 -Dio (30 mg/m 2 /day if age 60) or e mitoxantrone 12 mg/m 2 i.v. Di-D 3 as a daily bolus or e idarubicin 13 mg/m 2 i.v. Di-D 3 as a daily bolus 1.2.2. Treatment of adults below 6) years of age i) induction chemotherapy: 15 This induction cycle incorporates the administration of cytarabine in a high dose according to the following scheme: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-Dio infusion of 1 h - 49 * Ara-C 2000 mg/m 2 /day i.v. Di-D, (cytarabine) as an infusion of 2 hours, every 12 hours e daunorubicin 60 mg/m 2 /day i.v. D 4

-D

6 as a continuous infusion of 24 hours or * cytarabine 3000 mg/m 2 /day i.v. D 1

-D

6 as an infusion of 1 hour, every 12 hours e daunorubicin 45 mg/m 2 bolus every i.v. Dy-D, 24 hours (in order to reduce the risk of S.N.S. toxicity, in the event of renal insufficiency, adjust the cytarabine dosage to the clearance 5 of creatinine) according to L.E. Damon et al. (Leukemia 1994; 8: 535-541), G.L. Phillips et al. (Blood 1991; 77: 1429-1435) and G. Smith et al. (J. Clin. Oncol. 1997; 15: 833-839). 10 ii) consolidation chemotherapy: The cycle, described below, will be repeated 8 times, at the rate of 1 cycle every 4 to 6 weeks (according to R.J. Mayer et al., 15 N. Engl J. Med. 1994; 331: 896-903): Dose Route Days " isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-Ds infusion of 1 h I I _I - 50 * cytarabine 3000 mg/m 2 i.v. D 1 , D 3 , D 5 as an infusion of 3 hours, every 12 hours (4 cycles) then 100 mg/m 2 /day s.c. Di-D5 e cytarabine every 12 hours e daunorubicin 45 mg/m 2 bolus i.v. Di (4 cycles) iii) consolidation chemotherapy (with a high dose of cytarabine): The cycle, described below, will have to be 5 repeated twice and is adapted according to G.L. Phillips et al, (Blood 1991; 77: 1429-1435) ; S.N. Wolff et al, (J. Clin. Oncol. 1989; 7: 1260-1267); R.J. Mayer et al. (N. Engl J. Med. 1994; 331: 896-903): 10 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-Dio infusion of 1 h e cytarabine 3000 mg/m 2 i.v. Di-D6 1 hour every 12 hours e daunorubicin 30-45 mg/m 2 /day i.v. D 7 -D bolus once/day 1.2.3. Treatment of adults aged 60 or above The claimed substances may be added to the 15 consolidation chemotherapy protocols below: i) according to R.O. Dilman et al, (Blood 1991; 78; 2520-2526), Z.A. Arlin et al. (Leukemia 1990; 4: 177-183), P.H. Wiernik et al. (Blood 20 1992; 79: 313-319): - 51 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 6 infusion of 1 h * cytarabine 100-200 mg/m 2 i.v. Di-Ds continuous infusion of 24 hours * daunorubicin 30-45 mg/m 2 /day i.v. D 1 , D 2 , bolus or * mitoxantrone 12 mg/m 2 /day i.v. D 1 , D 2 bolus or * idarubicin 13 mg/m 2 /day i.v. D 1 , D 2 bolus ii) according to R.J. Mayer et al. (N. Engl. J. Med, 194; 331: 896-903): 5 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D, infusion of 1 h * cytarabine 100 mg/m 2 i.v. Di-Ds continuous infusion of 24 hours (4 cycles) then * cytarabine s.c. Di, D 5 100 mg/m 2 every 12 hours * daunorubicin 45 mg/m 2 /day i.v. Di bolus (4 cycles) - 52 iii) according to C.A. Linker et al. (Blood 1993; 81: 311-318), N. Chao et al. (Blood 1993; 81: 319-323) and A.M. Yeager et al. (N. Eng. J. Med. 1986; 315: 145-147): 5 This protocol comprises an autologous bone marrow transplant (performed on day Do): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v.

D_

7 -D-2 infusion of 1 h e busulfan 1 mg/kg qid oral D-7 to D 4 (in total 16 doses) e etoposide 60 mg/kg/day i.v.

D

3 infusion of 10 hours 10 or Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D_ 9

-D

1 infusion of 1 h e busulfan 1 mg/kg qid oral D_ 9 to D-, * cyclo- 50 mg/kg/day i.v.

D-

5 to D-2 phosphamide infusion of 1 hour iv) in the case of HLA-compatible allogeneic bone marrow transplant according to: 15 P.J. Tutscha et al. Blood 1987; 70: 1382-1388, F.R. Applebaum et al., Ann. Int. Med. 1984; 101: 581-588: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D-7-D infusion of 1 h - 53 * busulfan 1 mg/kg qid oral D- 7 to D 4 (in total 16 doses) e cyclo- 60 mg/kg/day i.v.

D

3 to D-2 phosphamide infusion of 1 hour 2/ Chronic adult leukaemias 2.1 Chronic myeloid leukaemia 5 In the myeloblastic phase, the isoflavonoids may be added to the HU-Mith treatment, described by C.A. Koller et al. (N. Engl. J. med. 1986; 315: 1433-1438): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or S - 50 mg/kg/day i.v. Di-Ds infusion of 1 h D8-Di2

D

15 -Di 9

D

2 2

-D

26 * hydroxyurea 500 mg/day oral every day * mithramycin 25 pg/kg/day i.v. daily for infusion of 2-4 3 weeks then hours 3 times/week 10 2.2 Chronic lymphocytic leukaemia 2.2.1 FCG-CLL protocol The isoflavonoids may be added to the "pulsed 15 chlorambucil" combinations as described by E. Kimby et al. (Leuk. Lymphoma 1991; 5 (Suppl.) 93-96) and by FCGCLL (Blood 1990; 75: 1422-1425): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-Ds, infusion of 1 h D-8-D-12,

D

15

-D

2 2 - 54 e chlorambucil 0.1 mg/kg/day oral once/day or e chlorambucil 0.4 mg/kg/day oral Di every 14 days and & prednisone 75 mg/day oral

D

1

-D

3 2.2.2 Fludarabine-CdA protocol according to H.G. Chun et al. (J. Clin. Oncol. 1991; 9: 175-188), M.J. Keating et al. (Blood 1989; 74: 5 19-25 / J. Clin. Oncol. 1991; 9: 44-49) and A. Saven et al. (J. Clin. Oncol. 1995; 13: 570-574): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 8 infusion of 1 h (once/month for 6 to 12 cycles) * fludarabine 25-30 mg/m 2 /day i.v. Di-D 5 infusion of 30 minutes [every 4 weeks for 6 to 12 cycles] or e cladibrine 0.09 mg/kg/day as a i.v. Di-D 7 continuous infusion [1 cycle every 28 to 35 days for 1 to 9 cycles (median: 4 cycles)] - 55 3/ Lymphoproliferative diseases 3.1 Hodgkin's disease 5 The isoflavonoids may be incorporated into the polychemotherapy protocols conventionally used for the treatment of Hodgkin's lymphoma: 3.1.1 AVDB protocol 10 according to G. Bonnadonna et al. (Cancer Clin. Trials 1979; 2: 217-226) and G.P. Canellos et al. (N. Engl. J. Med. 1993; 327: 1478-1484): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D 3 , or 5 - 50 mg/kg/day i.v. D 15

-D

18 infusion of 1 h e doxorubicin (A) 25 mg/m 2 bolus i.v. D 1 , D 15 e bleomycin (B) 10 U/m 2 bolus i.v. D 1 , D 15 e vinblastine (V) 6 mg/m 2 bolus i.v. Di, D 15 e dacarbazine (D) 375 mg/m 2 bolus i.v. D 1 , D 15 the cure comprising 6 to 8 cycles, at the rate 15 of 1 cycle every 28 days. 3.1.2 MOPP/ABVD protocol according to G. Bonnadonna et al. (Ann. Intern. Med. 1986; 104: 739-746) and G.P. Canellos et al. 20 (N. Engl. J. Med. 1993; 327: 1478-1484): The MOPP protocol should be alternated with the ABVD protocol (cf. § 3.1.1) every 28 days and the cure comprises 6 cycles: MOPP protocol: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. D 1

-D

3 , infusion of 1 h D 8 -Du 1 and

D

14

-D

17 - 56 * mechlorethamine 6 mg/m 2 bolus i.v. D 1 , D 8 (M) e vincristine (0) 1.4 mg/m 2 bolus i.v. DI, D 8 (no maximum) * procarbazine (P) 100 mg/m 2 /day oral Di-D 14 1 prednisone (P) 40 mg/m 2 /day oral Di-D 14 3.1.3 Stanford V protocol according to N.L. Bartlett et al. (J. Clin. Oncol. 1995; 13: 1080-1088): 5 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D 5 or 5 - 50 mg/kg/day i.v. Ds-Di2 infusion of 1 h D 15 -Dig

D

2 2

-D

2 6 e doxorubicin 25 mg/m 2 i.v. D 1 , D 15 e vinblastine 6 mg/m 2 bolus i.v. Di, D 15 (4 mg/m 2 during cycle 3 if age > 50) * mechlorethamine 6 mg/m 2 bolus i.v. Di (M) e vincristine 1.4 mg/m 2 bolus i.v. D 1 , D 22 (max. dose: 2 mg) [1 mg/m 2 during cycle 3 if age > 50) * bleomycin 5 U/m2 i.v. D 8 , D 2 2 * etoposide 60 mg/m 2 oral D 15 , D 16 * prednisone 40 mg/m 2 /day oral once/week (weeks 1-9) the cure comprising 3 cycles, at the rate of 1 cycle every 28 days. 3.1.4 EVA protocol - 57 according to G.P. Canellos et al. (Proc. Am. Soc. Clin. Oncol. 1991; 10: 273): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D, or 5 - 50 mg/kg/day i.v. infusion of 1 h " etoposide (E) 100 mg/m 2 infusion oral D 1 , D 2 , D 3 of 2 hours e vinblastine (V) 6 mg/m 2 bolus i.v. D, e doxorubicin (A) 50 mg/m 2 bolus i.v. Di the cure comprising 6 cycles, at the rate of 5 1 cycle every 28 days. 3.1.5 B-CAVe protocol according to W.G. Harker et al. (Ann. Intern. Med. 1984; 101: 440-446): 10 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D3 or 5 - 50 mg/kg/day i.v. infusion of 1 h * bleomycin (B) 5 U/m 2 bolus i.v. Di e lomustine (CCNU) 100 mg/m 2 oral D, e doxorubicin (A) 60 mg/m 2 bolus i.v. D e vinblastine (Ve) 5 mg/m 2 bolus i.v. the cure comprising 8 cycles, at the rate of 1 cycle every 28 days. 3.2. Non-Hodgkin's lymphomas 15 3.2.1. of low grade of malignancy i)-CVP protocol - according to C.M. Bagley et al. (Ann. Intern. Med. 1972; 76: 227-234) and C.S. Portlock et 20 al. (Blood 1976; 47: 747-756) - 58 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D 5 or 5 - 50 mg/kg/day i.v. infusion of 1 h e cyclophosphamide 300-400 mg/m 2 /day oral D 1 , D 5 (c) e vincristine (v) 1.4 mg/m 2 bolus i.v. Di (max: 2 mg) * prednisone (P) 100 mg/m 2 day oral Di-D 5 This cycle is repeated every 21 days up to the maximum response 5 ii)- I-COPA protocol - according to RV Smalley et al. (N. Eng. J. Med. 1992; 327: 1336-1341) Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D 5 or 5 - 50 mg/kg/day i.v. infusion of 1 h e cyclophosphamide 600 mg/m 2 /day i.v. Di (C) * vincristine (0) 1.2 mg/m 2 bolus i.v. Di (max: 2 mg) e prednisone (P) 100 mg/m 2 /day i.v. Di-D, e doxorubicin (A) 50 mg/m 2 bolus i.v. Di * interferon-alpha 6 MU/m 2 i.m. D 2 2

-D

2 6 (I) I I The cure comprises 8 to 10 cycles, at the rate 10 of one cycle every 28 days. iii)- Fludarabine-CdA protocol - according to P. Solol-Celigny et al. (Blood 1994; 84 (Supp. 1): 383a), H. Hoeschster et 15 al.; (Blood 1994; 84 (Suppl. 1): 564a and A.C. Kay (J. Clin. Oncol. 1992; 10: 371-377) - 59 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D, infusion of 1 h e fludarabine 25 mg/m 2 /day i.v. Di-Ds infusion of 0.5 hour or * fludarabine 20 mg/m 2 /day i.v. Di-Ds and cyclophos- 600 - 1000 mg/m 2 /day i.v. Di phamide or cladribine 0.1 mg/m 2 /day i.v. Di-D, infusion of 24 hours For fludaribine, each cycle is repeated every 28 days; for cladribine, each cycle is repeated every 35 days. 5 3.2.2. of intermediate malignancy grade i)-CHOP or CNOP protocol - according to EM McKelvey et al. (Cancer 1976; 10 38: 1484 - 1493), J.O. Armitage et al. (J. Clin. Oncol. 1984; 2: 898-902, S. Paulovsky et al. (Ann. Oncol. 1992; 3: 205-209) Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day i.v. Di-Ds or 5 - 50 mg/kg/day infusion of 1 h * cyclophosphamide 750 mg/m 2 /day i.v. Di (c) e doxorubicin (H) 50 mg/m 2 bolus i.v. Di * vincristine (0) 1.4 mg/m 2 bolus i.v. Di (max. 2 mg) - 60 e prednisone (P) 100 mg/m 2 /day (as oral Di-Ds 1 dose/day) for the CHOP protocol The mitoxantrone (N) may be used to replace (CNOP protocol) the doxorubicin in patients over 60 (dose: 12 mg/m2 as an i.v. bolus on day Dl of each 5 cycle) The cure by the CHOP or CNOP protocol comprises 6 to 8 cycles at the rate of 1 cycle every 21 days. ii) - MACOP-B protocol 10 - according to P. Klimo et al. (Ann. Intern. Med. 1985; 102: 596-602) and I.A. Cooper et al. (J. Clin. Oncol. 1994; 12: 769-778) Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D 5 , or 5 - 50 mg/kg/day i.v. D 8

-D

12 , infusion of 1 h Dis-D22,

D

2 9

-D

3 3

D

4 3

-D

4 7 ,

D

57

-D

61

D

71

-D

75 e methotrexate (M) 100 mg/m 2 /bolus then i.v. D 8 , D 36 , 300 mg/m 2 infusion

D

6 4 of 4 hours * leucovorin 15 mg qid oral D 9 , D 37 , D65 e doxorubicin (A) 50 mg/m 2 bolus i.v D 1 , D 15 ,

D

2 9 , D 4 3 ,

D

57 , D 71 e cyclo- 350 mg/m 2 bolus i.v. D 1 , D 5 , phospshamide (c)

D

2 9 , D 4 3 ,

D

57 , D 7 1 e vincristine (D) 1.4 mg/m 2 bolus i.v. D 8 , D 2 2 , (max: 2 mg)

D

36 , D 50 ,

D

6 4 , D 78 - 61 * prednisone (P) 75 mg/day oral Every day for 12 weeks * bleomycin 10 U/m 2 bolus i.v. D 22 , D 50 , D 78 This treatment protocol extends over 12 weeks and corresponds to 1 cycle. iii) - VACOP-B protocol 5 - according to J.M. Connors et al. (Proc. Am. Soc. Clin. Oncol. 1990; 9:254): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D 5 , or 5 - 50 mg/kg/day i.v. D 8

-D

12 , infusion of 1 h Dis-D22

D

2 9

-D

34 ,

D

4 3

-D

4 7 ,

D

5 7

-D

61 ,

D

7 1

-D

75 e etoposide (V) 50 mg/m 2 i.v. D 15 , D 4 3 , D71 e etoposide 100 mg/m 2 oral D 16 , D 17 ,

D

44 , D 45 , D72, D73 e doxorubicin (A) 50 mg/m 2 bolus i.v. D 1 , D 15 ,

D

2 9 , D 4 3 , D57, D71 * cyclophosphamide 30 mg/m 2 day bolus i.v. D 8 , D 2 2 , (c) D 36

D

50 ,

D

64 , D 78 e vincristine (0) 1.2 mg/m 2 bolus i.v. D 8 , D 2 2 ,

D

36 , D 50 , D64, D 78 - 62 e prednisone (P) 45 mg/m 2 /day oral 1/day for 1 week, then 4/day the next 11 weeks Each cycle lasting for 12 weeks. iv)- m-BACOD/M-BACOD protocol - according to M.A. Shipp et al. (Ann. Int. 5 Med. 1986; 140: 757-765) and A.T. Skarin et al. (J. Clin. Oncol. 1983; 1:91-98) Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D 5 , or 5 - 50 mg/kg/day i.v. D 8

-D

12 , infusion of 1 h D 15 -Di, e methotrexate 200 mg/m 2 i.v D 8 , D 15 (M) infusion of 4 hours or or (M) 3000 mg/m 2 infusion i.v. D 15 of 4 hours e leucovorin 10 mg/m 2 qid oral D 9 , D 16 or (6 doses in total)

D

16 * bleomycin (B) 4 U/m 2 bolus i.v. Di e doxorubicin (A) 45 mg/m 2 day bolus i.v. D, * cyclophosphamide 600 mg/m 2 bolus i.v. D, (C) e vincristine (0) 1 mg/m 2 bolus i.v. Di e dexamethasone 6 mg/m 2 /day oral D3-Ds (D ) I I The cure comprising 10 cycles, at the rate of 10 1 cycle every 21 days. v) - ProMACE/CytaBOM protocol - according to D.L. Longo et al. (J. Clin. Oncol. 1991; 9: 25-38): - 63 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-D 5 , or 5 - 50 mg/kg/day i.v. D8-D12 infusion of 1 h * cyclophosphamide 650 mg/m 2 infusion i.v. Di (C) of 0.5 hour e doxorubicin (A) 25 mg/m 2 bolus i.v. Di e etoposide 120 mg/m 2 infusion i.v. Di of 1 hour e prednisone (P) 60 mg/day oral Di-D 14 * cytarabine 300 mg/m 2 bolus i.v D8 e bleomycin (B) 5 U/m 2 bolus i.v D, * vincristine (0) 1.4 mg/m 2 bolus i.v D, * methotrexate 120 mg/m 2 bolus i.v D, * leucovorin 25 mg/m 2 qid oral D 9 (4 doses in total) The cure comprising 6 to 8 cycles, at the rate of one cycle every 14 days. 5 3.2.3. of low or intermediate malignancy grade i)- ESHAP rescue protocol - in case of recidivation or in case of failure of the first line treatment, according to 10 W.S. Velasquez et al. (J. Clin. Oncol. 1994; 12: 1169-1176) Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day Di-Ds or 5 - 50 mg/kg/day i.v. infusion of 1 h * etoposide (E) 40 mg/M 2 infusion of i.v. Di-D4 2 hours * methyl- 500 mg/day infusion i.v. D 1 , D 4 prednisolone (S) of 15 minutes - 64 " cytarabine (HA) 2000 mg/m 2 infusion i.v. Ds of 3 hours e cisplatin (P) 25 mg/m 2 /day bolus i.v. Di-D4 continuous infusion I_ of 24 hours The cure comprising 6 cycles, at the rate of 1 cycle every 28 days. ii)- MINE rescue protocol 5 - in case of recidivation or in the case of failure of the first line treatment, according to F. Cabanillas et al. (Semin. Oncol. 1990; 17 (Suppl. 10): 28-33) Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D, infusion of 1 h e ifosfamide (I) 1330 mg/m 2 infusion i.v. Di-D3 of 1 hour * mesna (M) 1330 mg/m 2 i.v. Di-D 3 in the ifosfamide infusion then 266 mg/m 2 bolus 4 and 8 hours after each dose of ifosfamide * mitoxantrone (M) 8 mg/m 2 infusion of i.v. D 15 minutes e etoposide (E) 65 mg/m 2 /day i.v. Di-D 3 infusion of 1 hour 10 This cycle to be repeated every 21 days. 3.3. Non-Hodgkin's lymphomas: Burkitt' s lymphoma, small cell lymphoma, lymphoblastic lymphoma 15 3.3.1 Magrath protocol - 65 - The claimed products may be combined with the Magrath protocols according to the following schemes: 5 i) - cycle 1 - according to I.T. Magrath et al. (Blood 1984; 63: 1102-1111) Dose Route Days " isoflavonoid 200-2000 mg/m 2 /day Di-D 5 , or 5 - 50 mg/kg/day i.v. D 8

-D

12 infusion of 1 h * cytarabine 30 mg/m 2 intra- Di, D 2 , thecal D,, D, * cyclophosphamide 1200 mg/m 2 bolus i.v. Di e methotrexate 12.5 mg/m 2 intra- Dio (max: 12.5 mg) thecal e methotrexate 300 mg/m 2 /day i.v. Dio-Du infusion of 1 hour then 60 mg/m 2 /h infusion of 41 hours * leucovorin 15 mg/m 2 bolus qid i.v. to be (8 successive started doses) 42 hours after the start of the admini stration of metho trexate 10 ii) - cycles 2 to 15 - according to I.T. Magrath et al. (1984) also - 66 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day D 1

-D

5 or 5 - 50 mg/kg/day i.v. D 10 , D 11 infusion of 1 h e cytarabine 45 mg/m 2 intra- D1-D2 thecal (cycles 2 and 3) Di (cycles 4 and 6) e cyclophosphamide 1200 mg/m 2 bolus i.v. Di (C) e doxorubicin 40 mg/m 2 bolus i.v. Di " vincristine 1.4 mg/m 2 bolus i.v. Di (max: 2 mg) * methotrexate 12.5 mg/m 2 intra- D 3 , D 10 (max: 12.5 mg) thecal (cycles 2 and 3) Dio (cycles 4, 5, 6) " methotrexate 300 mg/m 2 infusion i.v. D 10 , D 11 of 1 hour then (cycles 2 60 mg/m 2 continuous and 6) infusion of D 14 , D 15 41 hours (cycles 7-15) e leucovorin 15 mg/m 2 bolus qid i.v. start at (8 consecutive the 42 nd doses) hour of the treatment with metho trexate the cure comprising 14 cycles, at the rate of one cycle every 28 days. 3.4 Waldenstr~m macroglobulinaemia 5 3.4.1 CVP protocol - 67 according to the CVP protocol described by M.A. Dimopoulous et al, (Blood 1994; 83: 1452-1459) and C.S. Portlock et al. (Blood 1976; 47: 747-756): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 infusion of 1 h * cyclo- 300-400 mg/m 2 /day oral Di-Ds phosphamide (C) * vincristine (V) 1.4 mg/m 2 /day bolus i.v. Di (max: 2 mg) * prednisone (P) 100 mg/m 2 /day oral Di-D 5 5 the cure to be continued indefinitely (1 cycle every 21 days). 3.4.2 Fludarabine-CdA protocol according to H.M. Kantarjian et al. (Blood 1990; 10 75: 1928-1931) and M.A. Dinopoulous et al. (Ann. Intern. Med. 1993; 118: 195-198): Dose Route Days * isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 infusion of 1 h e fludarabine 25-30 mg/m 2 infusion i.v. Di-D, of 0.5 hour or 15 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D, infusion of 1 h " cladribine 0.09 mg/m 2 /day i.v. Di-D, (CdA) continuous infusion the cure comprising 6 to 12 cycles 28 days apart in the case of fludarabine and 2 cycles 28 days apart also in the case of cladribine.

- 68 3.5 Multiple myeloma 3.5.1 MP protocol according to R. Alexanian et al. (JAMA 1969: 5 208: 1680-1685), A. Belch et al. (Br. J. Cancer 1988; 57: 94-99) and F. Mandelli et al. (N. Engl. J. med. 1990; 322: 1430-1434): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-Ds infusion of 1 h * melphalan (M) 0.25 mg/kg/day oral Di-D4 1 prednisone (P) 100 mg/day oral I _Di-D4 10 or Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D, infusion of 1 h * melphalan (M) 9 mg/m 2 /day oral Di-D4 e prednisone (P) 100 mg/day oral Di-D4 the cure comprising at least 12 cycles, at the rate of 1 cycle every 4 to 6 weeks. 15 3.5.2 VAD protocol according to B. Barlogie et al, (N. Engl. J. Med. 1984; 310: 1353-1356): Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D, infusion of 1 h " vincristine (V) 0.4 mg/day i.v. Di-D4 continuous infusion of 24 hours - 69 e doxorubicin (A) 9 mg/m 2 /day i.v. Di-D4 continuous infusion of 24 hours * dexamethasone 40 mg/day i.v. Di-D4, (D)

D

9

-D

12 ,

D

1 7

-D

2 0 3.5.3 MP-interferon a protocol according to 0. Osterborg et al. (Blood 1993; 81: 1428-1434): 5 Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-Ds infusion of 1 h e melphalan (M) 0.25 mg/kg/day oral Di-D4 e prednisone (P) 2 mg/kg/day oral Di-D 4 e interferon- 7 MU/m 2 /day s.c. Di-D 5 and alpha

D

22

-D

26 the cure comprising the indefinite repetition of this cycle, at the rate of 1 cycle every 42 days. 3.5.4 VCAP or VBAP protocol 10 according to S.E. Salmon et al. (J. Clin. Oncol. 1983; 1: 453-461): VCAP protocol: Dose Route Days e isoflavonoid 200-2000 mg/m 2 /day or 5 - 50 mg/kg/day i.v. Di-D 5 infusion of 1 h e vincristine (V) 1 mg/m 2 bolus (max: i.v. Di 1.5 mg) e doxorubicin 30 mg/m 2 bolus i.v. Di e prednisone (P) 60 mg/m 2 /day oral Di-D4 * cyclophosphamide 125 mg/m 2 oral Di-D 4 (C ) II__ VBAP protocol: the cyclophosphamide is replaced 15 with carmustine (BCNU), the remainder being identical: - 70 Dose Route Days e carmustine 30 mg/m 2 infusion of i.v. D, 1 hour C. CHILDHOOD TUMOURS - Paediatric oncology The isoflavonoids may also be incorporated into 5 the polychemotherapy protocols for treating paediatric tumours in order to enhance the antitumour efficacy while reducing the severity of the side effects by means of the action on the recruitment and mobilization of clonogenic cells and the possibility of reducing the 10 active doses. 1/ Ewing sarcoma/primitive neuroectodermal tumour The isoflavonoids may be introduced in the VCR-Doxo-CY-Ifos-Mesna-E (E.D. Bergert et al., J. Clin. 15 Oncol. 1990; 8: 1514-1524; W.H. Meyer et al., J. Clin. Oncol. 1992; 10: 1737-1742): Dose Route Days e isoflavonoid 100-200 mg/m 2 /day or Di-D 5 , 2 - 50 mg/kg/day i.v. Ds-Du, infusion of 1 h Dis-Dis,

D

2 2

-D

2 7 , * vincristine 2 mg/m 2 bolus i.v. Di, D 8 , D 15 , (maximum dose =D43 2 mg) e doxorubicin 30 mg/m 2 /day i.v. Di-D 3 , as an infusion of D 4 3

-D

45 24 hours e cyclo- 2.2 g/m 2 as an i.v. 'Di, D 4 3 phosphamide infusion of 0.5 hour " ifosfamide 1800 mg/m 2 /day as an i.v. D22-D26 infusion of 1 hour D 63

-D

67 - 71 e mesna 360 mg/m 2 as an i.v. admini infusion of stered with 15 minutes at the cyclophos rate of 5 doses phamide and every 3 hours ifosfamide * etoposide 100 mg/m2 as an i.v. D 2 2

-D

26 infusion of 1 hour

D

63

-D

67 the cure comprises 6 to 10 of these cycles depending on the initial severity of the sarcoma and the extent of the response. 5 2/ Childhood acute lymphoblastic leukaemia 2.1. Induction chemotherapy (days D-Do 30 ) The isoflavonoids may be added to the recommended protocols (P.S. Gaynon et al., J. Clin. Oncol., 1993, 11, 2234-2242; J. Pullen et al., J. Clin. 10 Oncol. 1993; 11: 2234-2242; J. Pullen et al., J. Clin. Oncol. 1993; 11: 839-849; VJ Land et al., J. Clin. Oncol. 1994; 12: 1939-1945): Dose Route Days e isoflavonoid 100-200 mg/m 2 /day or Di-D 5 and 2 - 50 mg/kg/day i.v. D 2 2

-D

2 7 and infusion of 1 h

D

1 , Da,

D

15 and

D

2 2 e vincristine 1.5 mg/m 2 bolus i.v. Di, D 8 , D 15 , (maximum dose =

D

2 2 2 mg) e L-asparaginase 6000 IU/m 2 i.m. 3 times/week for 3 weeks e prednisone 60 mg/m2 in oral Di to D 2 8 3 doses/day * daunorubicin 25 mg/m 2 day as an i.v.

D

1 , D 8 , D 15 infusion of and D 2 2 15 minutes * methotrexate depending on age intra-

D

15 , D 2 8 the thecal - 72 cytarabine depending on age intra- Di thecal depending on the result of the examination of the bone marrow, the passage to the consolidation phase is made on day D 28 of the treatment protocol. 5 2.2. Consolidation/maintenance chemotherapy The isoflavonoids may be introduced in the maintenance protocol (P.S. Gaynon et al., J. Clin. Oncol. 1993; 11: 2234-2242; J. Pullen et al., J. Clin. Oncol. 1993; 11: 839-849; V.J. Land et al., J. Clin. 10 Oncol. 1994; 12: 1939-1945) according to the following scheme: Dose Route Days e isoflavonoid 100-200 mg/m 2 /day Di-D 5 , D 15

-D

2 0 or i.v. and D 94

-D

99 , 2 - 50 mg/kg/day Dioi-Dios, infusion of 1 hour

D

108

D

11 3 ,

D

12 2

-D

1 2 7 e cyclophos- 1000 mg/m2 as an i.v. Di, D 1 5 , D 1 2 2 phamide infusion of 0.5 hour * L-asparaginase 6000 U/m 2 i.m. 3 times/week between

D

9 7 and D1 2 2 * cytarabine 75 mg/m 2 /day as an i.v./s.c. a sequence of infusion of 4 days 15 minutes starting D 2 , Ds, D 16 , D 23 ,

D

1 2 3 , D, e doxorubicin 25 mg/m 2 /day as an i.v. D 94 , D 101 , D 108 infusion of 15 minutes e mercaptopurine 60 mg/m 2 /day oral Di-D 93 , D 14 3 at the end of the treatment - 73 e methotrexate 20 mg/m 2 /day oral once/week between

D

36 and D 72 and between D 143 and the end of the treatment * prednisone 40 mg/m 2 /day oral 5 consecutive (divided into days per 3 doses/day) month between

D

143 and the end of the treatment * thioguanine 60 mg/m 2 /day oral D 1 2 2

-D

1 35 e vincristine 1.5 mg/m 2 bolus i.v. D 94 , Dioi, D 108 , (maximum dose = then 2 mg) once/month between D 143 and the end of the treatment " methotrexate depending on age intra- D 1 , D 8 , D 15 , thecal D 2 2 , D 1 2 3 , D 1 3 0 then once/3 months between

D

143 and the end of the treatment 3/ Childhood acute myeloid leukaemia The isoflavonoids are added to the induction and consolidation/maintenance protocols according to 5 the following schemes: 3.1. Induction chemotherapy According to Y. Ravindranath et al., J. Clin. Oncol. 1991; 9: 572-580; M.E. Nesbit et al., J. Clin.

- 74 Oncol. 1994; 12: 127-135; RJ Wells et al., J. Clin. Oncol. 1994; 12: 2367-2377): Dose, Route Days e isoflavonoid 100-200 mg/m 2 /day or i.v. Di-D 5 , 2 - 50 mg/kg/day

D

10

-D

13 infusion of 1 h * cytarabine according to age intra- Di thecal e daunorubicin 20 mg/m 2 /day as an i.v. Di-D4, infusion of Dio-Dia 24 hours * cytarabine 200 mg/m 2 /day as an i.v. Di-D4, infusion of

D

1 0

-D

13 24 hours * thioguanine 100 mg/m 2 /day oral Di-D 4 , divided into

D

10

-D

13 2 doses/day e etoposide 100 mg/m 2 /day as an i.v. Di-D4, infusion of

D

1 0

-D

1 3 24 hours e dexamethasone 6 mg/m 2 divided i.v./ Di-D4, into 3 doses/day oral Dio-D 13 this cycle being repeated from D 28 . 5 3.2. Consolidation/maintenance chemotherapy According to Y. Ravidranath et al., J. Clin. Oncol. 1991; 9: 572-580; M.E. Nesbit et al., J. Clin. Oncol. 1994; 12: 127-135; R. J. Wells et al, J. Clin. 10 Oncol. 1994; 12: 2367-2377): Dose Route Days e cytarabine according to age intra- D 1 , D 2 8 , D 56 thecal " isoflavonoid 100-200 mg/m 2 /day Dj-D 5 , D 8

-D

13 or i.v. and D 2 8

-D

33 , 2 - 50 mg/kg/day D56-D6i infusion of 1 h

D

8 9

_D

9 4 - 75 e cytarabine 3000 mg/m2 as an i.v. D 1

-D

2 , and infusion of D 8 -D 3 hours every 12 hours " L-asparaginase 6000 IU/m 2 i.m. D 2 , D 9 3 hours after cytarabine * vincristine 1.5 mg/m 2 bolus i.v. D 2 8 , D 56 (maximum dose = 2 mg) * thioguanine 75 mg/m 2 /day oral D 28

-D

8 4 e cytarabine 25 mg/m 2 /day bolus i.v. D28-D3,

D

56

-D

59 e cyclophos- 75 mg/m 2 /day as an i.v. D23-Du, phamide infusion of 0.5 D 56

-D

59 hour * cytarabine 25 mg/m 2 /day bolus sc/i.v D 89

-D

93 * thioguanine 50 mg/m 2 /day oral D 8 9

-D

93 " etoposide 100 mg/m 2 /day as an i.v. Da 9 , D 9 2 infusion of 1 hour e dexamethasone 2 mg/m 2 /day oral D 8 9

-D

92 e daunorubicin 30 mg/m 2 as an i.v. D 89 infusion of 15 minutes 4/ Childhood Hodgkin's disease The isoflavonoids may be addeed to the MOPP-ABVD protocol according to EA Gehan et al. (Cancer 5 1990; 65: 1429-1437), SP Hunger et al. (J. Clin. Oncol. 1994; 12: 2160-2166) and MM Hudson et al. (J. Clin. Oncol. 1993; 11: 100-108): Dose Route Days e isoflavonoid 100-200 mg/m 2 /day or i.v. Di-Ds and 2 - 50 mg/kg/day Ds-D 12 infusion of 1 h - 76 * mechlorethamine 6 mg/m 2 bolus i.v. Di, D 8 (M) e vincristine (0) 1.5 mg/m 2 bolus i.v. D 1 , D 8 (maximum 2 mg) e procarbazine 100 mg/m 2 /day oral D1-Da (P) e prednisone (P) 40 mg/m 2 /day oral Di-D1 (divided into 3 doses/d) e doxorubicin (A) 25 mg/m 2 /day as an i.v. D 2 9 , D 43 infusion of 15 minutes e bleomycin (B) 10 U/m 2 as an i.v. D 2 9 , D 4 3 infusion of 15 minutes e vinblastine 6 mg/m 2 bolus i.v. D 2 9 , D 4 3 (maximum 2 mg) e dacarbazine (D) 375 mg/m 2 as an i.v. D 2 9 , D 4 3 infusion of 15 minutes This cycle should be repeated 6 times at the rate of 1 cycle every 8 weeks, the cure comprising 6 cycles. If an autologous bone marrow transplant 5 (autograft) is prescribed, the CVB protocol described by R. Chopra et al. (Blood 1993; 81: 1137-145), C. Wheeler et al. (J. Clin. Oncol. 1990; 8: 648-656) and RJ Jones et al (J. Clin Oncol 1990, 8, 527-537) may be used according to the following scheme (the 10 allograft taking place on day Do): Dose Route Days e isoflavonoid 100-200 mg/m 2 /day or i.v. D- 7 , D_ 1 2 - 50 mg/kg/day infusion of 1 h * cyclo- 1800 mg/m 2 /day as i.v. D- 7 , D- 6 phosphamide 2 infusions of D- 5 , D- 4 1 hour - 77 e carmustine 112 mg/m 2 /day as an i.v. D- 7 , D- 6 (BCNU) infusion of 0.5 D-5, D 4 hour e etoposide 500 mg/m 2 /day as i.v. D- 7 , D.

6 2 infusions of

D-

5 , D 4 1 hour 5/ Childhood lymphoblastic lymphoma The isoflavonoids may also be combined with the induction chemotherapy protocols (A.T. Meadows et al., 5 J. Clin. Oncol. 1989; 7: 92-99 - C. Patte et al., Med. Ped. Oncol. 1992; 20: 105-113 and A. Reiter et al., J. Clin. Oncol. 1995; 13: 359-372) and the maintenance chemotherapy protocols: 10 5.1 Induction chemotherapy Dose Route Days e isoflavonoid 100-200 mg/m 2 /day or i.v. Di-D 5 , D 1 7

-D

2 2 , 2 - 50 mg/kg/day

D

2 4

-D

2 9 infusion of 1 h * cyclo- 1200 mg/m2 as an i.v. Di phosphamide infusion of 0.5 hour e cytarabine according to age intra- Di thecal * vincristine 1.5 mg/m 2 bolus i.v. D 3 , D 10 , D 1 7 , (maximum 2 mg) D24 e prednisone 60 mg/m 2 /day oral

D

3

-D

28 divided into 3 doses/day * daunorubicin 60 mg/m 2 i.v. D17 as an infusion of 15 minutes * L-asparaginase 6000 U/m 2 /day im

D

1 7

-D

35 as an infusion of 3 times/week 15 minutes - 78 * methotrexate according to age intra- D 17 , D 31 thecal 5.2 Maintenance chemotherapy according to the following scheme: Dose Route Days e isoflavonoid 100-200 mg/m 2 /day or i.v. Di-D 5 , D 1 5

-D

2 0 , 2 - 50 mg/kg/day

D

2 9

-D

3 4 infusion of 1 h e cyclo- 1000 mg/m2 as an i.v. Di phosphamide infusion of 0.5 hour * vincristine 1.5 mg/m 2 bolus oral Di, D 5 , (maximum 2 mg) (cycles 2 to 10) " methotrexate 300 mg/m 2 /day (60% i.v. Dis as an infusion of 15 minutes and 40% as an infusion of 4 hours) e leucovorin 10 mg/m 2 /every 4 h oral D1 " daunorubicin 30 mg/m 2 i.v.

D

2 9 as an infusion of 0.5 hour " methotrexate according to the intra- Di, D 8 , D 15 age thecal (cycle 1), then once/month (cycles 2 to 10) 5 the cure comprising 10 cycles 6/ Paediatric neuroblastoma The recommended polychemotherapy Doxo-E-Cy-Pt protocol is adapted from R.P. Castleberry et al. 10 (J. Clin. Oncol. 1992; 10: 1299-1304), A. Garaventa et - 79 al. (J. Clin. Oncol. 1993; 11: 1770-1779) and D.C. West et al. (J. Clin. Oncol. 1992; 11: 84-90): Dose Route Days e isoflavonoid 100-200 mg/m 2 /day or i.v. Di-D 5 , D 28

-D

35 , 2 - 50 mg/kg/day

D

58

-D

65 infusion of 1 h e doxorubicin 25 mg/m 2 /day as i.v. D 2 , D 3 0 , D 58 an infusion of 15 minutes e etoposide 100 mg/m 2 as an oral/ D 2 , D 5 , D 30 , infusion of naso- D 33 , D 58 , D 61 1 hour gastric " cyclo- 1000 mg/m2 as a i.v. D 3 , D 4 , D 31 , phosphamide infusion of 0.5

D

32 , D 59 , D 60 hour e cisplatin 60 mg/m 2 as an i.v. Di, D 28 , D 56 infusion of 6 hours The evaluation of the therapeutic response is 5 made after 9 weeks in order to decide on the attitude: surgical resection, radiotherapy or new chemotherapy. 7/ Paediatric osteosarcoma The isoflavonoids may be added to the 10 Doxo-Pt-Mtx-Lcv protocol as described by M. Hudson et al. (J. Clin. Oncol. 1990; 8: 1988-1997), PA Meyers (J. Clin. Oncol. 1992; 10: 5-15), and V.H.C. Bramwell et al. (J. Clin. Oncol. 1992; 10: 1579-1591): Dose Route Days e isoflavonoid 100-200 mg/m 2 /day or i.v. Di-D 5 , D 21

-D

26 , 2 - 50 mg/kg/day

D

2 8

-D

33 infusion of 1 h e doxorubicin 25 mg/m 2 /day as an i.v. Di-D 3 infusion of 24 hours - 80 e cisplatin 120 mg/m2 as an i.v. Di infusion of 6 hours e methotrexate 12 mg/m 2 /day as an i.v. D 21 , D 28 infusion of 1 hour e leucovorin 100 mg/m 2 oral D 2 2 , D 2 9 every 6 hours 8/ Childhood rhabdomyosarcoma The Vcr-Dact-CY-Mesna protocol (H. Maurer et al., Cancer 1993; 71: 1904-1922 and LR Mandell et al., 5 Oncology 1993; 7: 71-83) may include i.v. infusion of the isoflavonoids according to the following scheme: Dose Route Days e isoflavonoid 100-200 mg/m 2 /day or i.v. Di-D 5 , DB-D 12 , 2 - 50 mg/kg/day D22-D27, infusion of 1 h D43-D47 e vincristine 1.5 mg/m 2 /day i.v. D 1 , Da, D 15 , (bolus maximum

D

22 , D 2 9 , D 36 , 2 mg)

D

43 , D 50 , D 57 e dactinomycin 0.015 mg/kg bolus i.v. Di-D 5 , D 2 2

-D

2 7 , (max daily dose:

D

4 3

-D

4 7 0.5 mg * cyclo- 2.2 g/m 2 as an i.v. D 1 , D 2 2 , D 4 3 phosphamide infusion of 1 hour e mesna 360 mg/m 2 as an i.v. D 1 , D 2 2 , D 4 3 infusion of 1 hour every 3 hours for 5 doses At the end of the 9th week of treatment, the efficacy should be evaluated in order to decide on the 10 future course of action (surgery, radiotherapy, continuation of the chemotherapy).

- 81 9/ Childhood Wilms tumour In the Vcr-Dact protocol as described by GJ D'Angio et al. (Cancer, 1989; 64: 349-360) and DM Green et al. (J. Clin. Oncol. 1993; 11: 91-95): 5 Dose Route Days e isoflavonoid 100-200 mg/m 2 /day or i.v. Di-D 5 , D 8

-D

12 , 2 - 50 mg/kg/day then every infusion of 1 h week e vincristine 2 mg/m 2 bolus (max i.v. D 7 then every dose: 2 mg) week " dactinomycin 0.045 mg/kg bolus i.v. D 1 , then (P 30 kg) every 3 weeks 1.35 mg/m 2 (P>30 kg) (max dose: 3 mg This protocol being started after the surgical resection. In case of autologous bone marrow transplant (autograft) according to A. Garaventar et al. (Med. 10 Pediatr. Oncol. 1994; 22: 11-14), the E-Thio-Cy protocol may be modified as follows Dose Route Days e isoflavonoid 100-200 mg/m 2 /day or i.v. D_-D_1 2 - 50 mg/kg/day infusion of 1 h e etoposide 1800 mg/m 2 i.v. D-8 (infusion of 24 hours) * thiotepa 300 mg/m 2 /day as an i.v. D- 7 , D- 6 , D- 5 infusion of 2 hours e cyclo- 50 mg/kg/day as an i.v. D_ 4 , D- 3 , D- 2 , phosphamide infusion of D_ 1 1 hour' - 82 the bone marrow transplant taking place on Do.

Claims

1. Composition having an activity on the proliferation of clonogenic cells in tumours and which comprises a therapeutically effective quantity of an 5 isoflavonoid or of an analogue of the chromone type.

2. Composition according to Claim 1, in which the isoflavonoid is chosen from the compounds of formula: Ri R 2 0 R5 ' (0) R3 R6 R4 0 10 in which formula: - R 1 , R 2 , R 3 and R 4 are chosen, independently of each other, from H, OH, a Ci-C 4 alkoxy group, an -OCOR 7 group, R 7 being a Ci-C 4 alkyl group, at least one of the 15 substituents R 1 , R 2 , R 3 or R 4 being other than H and it being possible for R 2 and R 3 to form together a methylenedioxy group, - R 5 is chosen from H, OH, a C 1 -C 4 alkoxy group, an 0-glycosyl group and a cyclohexyl group, 20 - R 6 is chosen from a cyclohexyl group, a phenyl group and a phenyl group substituted 1 to 3 times with groups chosen from H, OH and a Ci-C 4 alkoxy group, - and - denotes either a double bond, or a single bond. 25

3. Composition according to Claim 2, in which the isoflavonoid is chosen from genistein, daidzein and biochanin A.

4. Use of an isoflavonoid or of an analogue of the chromone type for the manufacture of a medicament 30 intended to interfere with the generation of clonogenic - 2 cells in tumours during a treatment of these tumours with at least one cytotoxic agent.

5. Use of a compound chosen from the compounds of formula: 5 Ri R 2 O R5 -00 R3: R6 R4 O in which formula: - R 1 , R 2 , R 3 and R 4 are chosen, independently of each 10 other, from H, OH, a Ci-C 4 alkoxy group, an -OCOR 7 group, R 7 being a Ci-C 4 alkyl group, at least one of the substituents R 1 , R 2 , R 3 or R 4 being other than H and it being possible for R 2 and R 3 to form together a methylenedioxy group, 15 - R 5 is chosen from H, OH and a C 1 -C 4 alkoxy group, an 0-glycosyl group, and a cyclohexyl group, - R 6 is chosen from a cyclohexyl group, a phenyl group and a phenyl group substituted 1 to 3 times with groups chosen from H, OH and a C 1 -C 4 alkoxy group, 20 - and- denotes either a double bond, or a single bond, for the manufacture of a medicament intended to interfere with the generation of clonogenic cells in tumours during a treatment of these tumours with at 25 least one cytotoxic agent.

6. Use according to Claim 5, in which the compound of formula I is chosen from genistein, daidzein and biochanin A.

7. Method for the chemotherapeutic treatment of a 30 tumour in a patient with at least one cytotoxic agent, which comprises the administration, during the treatment with the cytotoxic agent, of a -3 therapeuticalkyl effective quantity of an isoflavonoid or of an analogue of the chromone type.

8. Method according to Claim 7, in which the isoflavonoid or analogue of the chromone type is 5 administered at the beginning of the chemotherapy treatment and at the beginning of each chemotherapy treatment cycle.