MXPA01000389A - Therapeutic composition based on flavonoids for use in the treatment of tumours with cytotoxic agents - Google Patents

Abstract

The invention concerns a composition having an activity on the proliferation of clonogenic cells in tumours and comprising a therapeutically efficient amount of an isoflavonoid or an analogous chromone compound, in particular a compound selected among the compounds of formula (I) wherein:R1, R2, R3 and R4 R5, and R6 are as defined in Claim 2. Said composition is designed for use in the treatment of tumours with cytotoxic agents.

Description

THERAPEUTIC COMPOSITION BASED ON ISOFLAVONOIDS INTENDED FOR USE IN THE TREATMENT OF TUMORS WITH AGENTS CITOTOXICS Field of the Invention The present invention relates to the use of compounds of the isoflavonoid type in the treatment of cancers with cytotoxic agents.

Background of the Invention A cancer is an alteration of the somatic genes in which, the genetic dysfunctions become amplified when the tumor process progresses from the state of a precancerous lesion to that of a malignant transformation, the cancerous tumor becomes metastatic and often resistant to cytotoxic drugs. Despite the major efforts made in all developed countries, particularly through clinical and experimental research programs, mortality due to various cancers (solid tumors and hematological malignancies) REF. DO NOT. 126272 remains unacceptably high. In many countries, mortality caused by cancer is classified as second, right after cardiovascular disease. 5 In terms of newly diagnosed cancers, the distribution between solid tumors and hematological malignancies (bone marrow, blood, lymphatic system) shows that 9 cancers out of 10 are solid tumors. Contrary to what is observed in hematologic oncology (therapeutic events in 40 to 90% of cancers of blood cells), only a smaller number of advanced or disseminated solid tumors respond to chemotherapy treatments alone. Is partially for this reason, that the total mortality caused by cancer increased in the United States between 1973 and 1992. It is undeniably undeniable that this trend can be reversed only by the The appearance, also of the arsenal of established chemotherapy of new antitumor drugs such as taxanes (paclitaxel and docetaxel), which interfere with the formation of microtubules (P. McGuire et al., Am. Intern. Med., 1989), the inhibitors of topoisomerases I derived from canfothecin (topotecano and irinotecano), vinorelbino (new alkaloids derived from vinvapervinca), gencitabine (new cytotoxic antimetabolic agents), raltitrexed (inhibitor of thymidylate synthetase) and miltefosina (first representative of the alkylphosphocholine family). These treatments are also, either as a first line treatment, or as a second line treatment, to drugs whose specific activity is now well recognized, such as doxorubicin, cisplatin, vincristine, methotrexate, 5-fluorouracil. One of the most difficult current problems of anticancer chemotherapy is due to the fact that many populations of malignant cells present substantial resistance to the established cytotoxic substances. More often, this situation results from the existence of multiple resistance genes or the frequency of genetic mutations in certain types of tumors. In addition, the treatment of cancer requires new scopes, complementary to those currently used, and proposed to better combat the extension and heterogeneity of the tumor burden and the acquisition of the resistance of the "multidrug drug". ^^^ 3 ¡^^^^^? J * g2 »Among these new scopes, some are already promising. This is the case for the induction of apoptosis, the inhibition of tumor angiogenesis and metastatic processes, without mention of gene therapy or immunotherapy. We inventors have been interested in a different scope. The suggested objective was to develop the population of the tumor cells most sensitive to the reference anticancer treatments in order to achieve a double beneficial effect: 1) increase the cytotoxic activity and, therefore, the efficacy, and 2) reduce the frequency and severity of certain side effects by virtue of the reduction of the dosage which could follow the induction of the increase in antitumor efficacy. In this strategy which is the origin of the discovery of an innovative mechanism caused by substances that have a low antitumor power or even that lack this power, but they are able to induce a very significant increase in the cytotoxic activity of the anticancer drugs tested. This innovative mechanism results from the possibility for these substances to either stimulate the recruitment of the clonogenic cells within the tumor, making them more sensitive to conventional treatment with cytotoxic agents, or inhibit the proliferation of clonogenic cells, thus contributing to tumor regression .

Description of the Invention The object of the present invention is furthermore, the use in the treatment of cancers with at least one antitumor agent selected from cytotoxic agents, of a compound having activity in the proliferation of clonogenic cells, selected from isoflavonoids and compounds analogs of the chromone type and in particular, the compounds of the formula: wherein, in the formula: Ri, R2, R3 and R4, are independently selected, from each of the others, of H, OH, a CX-C alkoxy group, a -OCOR group, R7 being an alkyl group C ~ C4 at least one of the substituents Ri R2 R3 or R4 are different from H and it is possible for R2 and R3 to together form a methylenedioxy group, R5 is selected from H, OH, a C4-C4 / 5 alkoxy group a group O-glycosyl and a cyclohexyl group, R5 is selected from a cyclohexyl group, a phenyl group and a phenyl group substituted 1 to 3 times with groups selected from H, OH and a C?-C4 alkoxy, 10 - and - ^ - group denotes either a double link, or a single link.

A preferred class of compounds of formula I are those in which R6 is selected from the phenyl group, the 4-hydroxy phenyl group and the 4- (C4-C4 alkoxy) phenyl groups. The cytotoxic agents can be used at their usual dose, in this case, their effectiveness is increased, or at lower doses, taking into account the increase in their antitumor efficacy if the desired objective is first, to increase the patient's tolerance to the treatment. The object of the present invention is also a composition having an activity in the proliferation of clonogenic cells by means of the < . , < lafe «« -. A > . "tea*- - . and y . and "and,. j,. "... .. "". Ü 3 j t ti SihSBtiíX. interference with the generation of clonogenic cells, either by the stimulation of proliferation and recruitment, or by the inhibition of proliferation, comprising a therapeutically effective amount of an isoflavonoid or an analogous compound of the chromone type, and in particular, of a compound selected from the compounds of formula: FU wherein, in the formula: Ri, R2, R3 and R4 are selected, independently of each other of H, OH, a C1-C4 alkoxy group, a group -OCOR7, R7 is a C-alkyl group? C4, at least one of the substituents Ri, R2, R3 or R4 are different from H and are possible for R2 and R3 to together form a methylenedioxy group, R5 is selected from H, OH, a C1-C4 alkoxy group, a group O-glycosyl, and a cyclohexyl group, R6 is selected from a cyclohexyl group, a phenyl group and a substituted phenyl group 1 to 3 times . t, .. $ & ' ^ t ^ ^ jü ^ A with selected groups of H, OH and an alkoxy group CI-C4, and - denotes either a double bond, or a single bond. The object of the present invention is also the use of an isoflavonoid, in particular of a compound of formula I as defined above, for the manufacture of a medicament proposed to interfere (by induction or inhibition) with the generation of clonogenic cells in tumors during a treatment with at least one cytotoxic agent. In the chemotherapeutic treatment of cancers with cytotoxic agents, the isoflavonoids and in particular the compounds of formula I, can be administered at the beginning of chemotherapy treatments either once, or several times a day at the beginning of these treatments (for example by 5 or 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 compounds of formula I, are advantageously administered by iion (generally during 1 a • "-" »* ^ - * - - * '3 hours) at a dose of 5 to 50 mg / kg / day or 200 to 2000 mg / m2 / day. To obtain a maximum effect on the production of clonogenic cells, the 5 isoflavonoids should be administered in such a way that the tissue concentrations obtained are the highest, which can be possibly contemplated. For the treatment protocols in the 10 acute phases of the cures, the intravenous route is referred to using: infusion solutions ready to be used (bags, flasks and the like) proposed to be administered as they are by intravenous infusion 15 with the help of a line of infusion and using the recommended flow rate: leophilized to be resuspended in solution by intravenous infusion with the help of pharmaceutical solutions known to persons skilled in the art; - for maintenance treatments, it is also possible to contemplate the oral route when the chemotherapy treatment preferably uses the administration of cytostatic agents by the oral route. For this purpose, lyophilized *-"*Y. ? fry. oral (for oral or perlingual absorption), tablets of instantaneous or delayed release, oral solutions, suspensions, granules, gelatin capsules and the like, can be used. The compounds of formula (I) are, for the most part, naturally occurring compounds or derivatives of naturally occurring compounds. As examples, there may be mentioned: 10-genistein, -biocanin A, daidzein, formononetin, 7-acetyl formononetin, 15-glycetein, orobol or 5, 7, 3 ', 4'-tetrahydroxy isoflavone, irizolone or 6,7-met ilendioxi- '-hydroxii soflavone, irigenin or 3', 5, 7-trihydroxy-4 ', 5', 6-methoxyisoflavone, 20-tectorigenin or 4 ', 5, 7-trihydroxy-6-methoxy soflavone, 2-hydroxy- 8-methoxy-2,3-dihydroisoflavone, 4 ', 7-dihydroxy-5-methoxyisoflavone.

Other isoflavones which can be ^^^ SÍ ^ ^ & tÉ ^^^ used are described by Donnelly et al. in Natural Product Reports, 1995, 321, or can be prepared by the methods described in this article. The cytotoxic agents may be selected from: i) intercalating agents, in particular doxorubicin, (adriamycin), daunorubicin, epirubicin, idarubicin, zorubicin, aclarubicin, pirarubicin, acridine, mitoxantrone, actinomycin D, eptilinium acetate; ii) alkylation agents selected from platinum derivative (cisplatin, carboplatin, oxaliplatin and the like), iii) a compound selected from the other groups of alkylating agents: cyclophosphamide, isfosfamide, chlorometryrin, melphalan, chlora bucyl, estramustine, busulfan, mitomycin C, - nitrosoureas: BCNU (carmustine), CCNU (lomustine), ustina fuse, streptozotocin, triazines or derivatives, procarbazine, dacarbazine, pipobroman, - ethylene glycine: altretamine, triethylenethiophosphoramide, iv) a compound selected from the other groups of antimetabolic agents : antifolic agents: methotrexate, raltitrexed, antipyrimidines: 5-fluorouracil (5-FU), cytarabine (Ara-C), hydroxyurea antipurins: purinetol, thioguanine, pentostatin, cladribine, inducers of cytotoxic nucleoside synthesis: gemcitabine, v) a compound selected from the other groups of agents with high affinity for the tubules: - vinca alkaloids which disorganize the mitotic axis: cincristine, vinblastine, vindesine, navelbine - agents that block the depolymerization of the mitotic axis: paclitaxel, docetaxel - agents that induce DNA breakdown by the inhibition of topoisomerase II: etoposide, teniposide - inhibitors of topoisomerase I that induce breaks in the DNA: topotecao, irinotecano, vi) a breaking agent, fragmentation of DNA such as bleomycin, vii) one of the following compounds: plicamycin, L-asparaginase, mitoguazone, dacarbazine, viii) an anticancer progestogenic steroid: medroxyprogesterone, magestrol, ix) an oestrogenic anticancer steroid: tetrasodium diethyl phosphodiestrol tetrasodium, x) an antiestrogen: tamoxifen, droloxifene, raloxifene, aminoglutethimide, xi) a steroidal antiandrogen (eg cyproterone example) or a non-steroidal antiandrogen (Flutamide, nilutamide).

In particular, the compounds of formula I can be combined with all treatments with the major cytotoxic agents used in polychemotherapy of solid tumors such as: - doxorubicin - alkylating agents: oxazoforins (cyclophosphamide, ifosfamide, chlorambucil, mephalan) - nitrosoureas - mitomycin C - antimerabolites such as methotrexate, 5-FU, . ? ? .á and Ara-C, capecitabine agents which interfere with tubilin: vinca alkaloids (vincristine, vinlastine, vindesine, navelbine), taxoids (paclitaxel, docetaxel), epidopofilotoxin derivatives (etoposide, teniposide), - bleomycin inhibitors of the topoisomerase I: topotecano, irinotecano. In the same way, the compounds of formula I can be combined with the treatment with the major cytotoxic agents used in oncohematology for the treatment of cancers blood: Hodgkin's disease: cyclophosphamide, mechlorethamine, chlorambucil, melphalan, ifosfamide, etoposide, dororubicin, daunorubicin; acute leukemias: methotrexate, 6- 20 mercaptopurine, cytarabine, vinblastine, vincristine, dororubicin, daunorubicin, L-asparaginase; Malignant non-Hodgkin lymphomas, mechlorethamine, chlorambucil, cyclophosphamide, melphalan, ifosfamide, methotrexate, cytarabine, vinblastine, vincristine, etoposide, doxorubicin, ^ ^ ^^^^^ t ^^^^^^^^^ CC ^^^^ C ^^^^^ s ^^^^^^^^^^^^^^^^ ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ g J ^ I daunorubicin, carmustine, lomustine, cisplatin; chronic lymphoid leukemia: mechlorethamine, chlorambucil, cyclophos, famide, melphalan, ifosfamide. The resulting pharmacological tests, which show the effects obtained, will be given below. 1- Interaction (stimulation or inhibition of proliferation) with the generation of clonogenic cells (clonogenic assay) The assay used is that described by Ha burger et al. (Science, 1977; 197, 461-463) and Salmon et al. (New England J. Med., 298, 1321-1327). A cell is considered to be clonogenic if it has the ability to proliferate and reach a cell colony. The "cells derived from human tumors" are the cells which are at the origin of the neoplastic cells that constitute a given tumor. These cells derived from the tumor are responsible for the recidivism process which can be observed after surgical resection of the primary tumors and are also responsible for the formation of metastases. At the level of a tumor or a tumor cell line, these clonogenic derived cells are distinguishable from the other tumor cells or the neoplastic cell line considered, by the fact that they can retain their ability to proliferate in the absence of some solid support. In this test, the tumor cells are cultured in a semi-solid support. Only the cells, which do not require a solid support for their growth (this is to say the highly tumorigenic cells called "ancorage-independent cells" by Ml Dawson et al., Cancer Res. 1995; 55: 4446-4451; so-called clonogenic cells with reference to "clonal growth") are capable of developing in such an agar-based support. Actually, in such a medium, normal cells - which grow in "adherent mode" ("ancorage-dependent cells" according to the terminology of M. I. Dawson) - such as, for example, fibroblasts, do not survive. Within a population of tumor cells, grown on such a support, these clonogenic cells (associated with an unlimited number of cell divisions and whose proliferation is called "ancorage-independent [clonal] growth" by MI Dawson), which are capable of grow. The percentage of these clonogenic cells within a tumor or a cell line varies between 0.1% and 0.001%. Non-clonogenic cells (associated with a limited number of cell divisions) do not develop in this test, because they require a solid support for their growth, which must occur in "adherent mode" ("ancorage dependent growth [adherent]" , according to MI Dawson et al., Cancer Res. 1995; 55: 4446-51). "The influence of compounds of formula (I) on the growth of the cell colonies obtained by cultivation was measured, for example, the lines of mammary tumor MCF7 and MXT and the HT-29 colorectal line in the semiliquid culture medium called "soft agar." In such medium, only the clonogenic cells called "independent ancorage cells (clonal)" by Ml Dawson survive and develop The growth of these cells in such a "non-adherent" way reflects their degree of tumorigenicity.The inhibition of the growth of the size of a tumor in which a large number of clonogenic cells have developed nces, reach control for a reinforced cytotoxic activity. For the contrast, this test also reveals that a compound is capable of inhibiting the generation / proliferation of clonogenic cells, which make the tumor less able to develop, and therefore, reduce the population of tumor cells. The tumor cell lines studied are maintained in the culture in falcon flasks of 25 cm2. They are then trypsinized and the cells well dissociated from each other. The percentage of living cells is determined after dyeing with trypan blue. A cell suspension at a concentration of 5,104 to 15,104 cells / ml (depending on the cell type considered) is prepared in a 0.3% agar solution. Then, 200 μl of this suspension is inoculated in 35 mm diameter Petri dishes, in which 3 mL of a lower layer consisting of a 0.5% agar solution is deposited. The 200 μl of the cell suspension are instead covered with 1.8 ml of an upper layer consisting of a 0.3% agar solution. The boxes are then placed in an incubator at 37 ° C, C02 at 5% and humidity at 70% until treatment. The latter is performed approximately 1 to 2 hours after inoculation. The compounds to be tested are prepared at a concentration of 100 parts greater than the desired concentration and 50 μl of these treatment solutions are deposited in the top layer of the corresponding boxes. In the present study, the final concentration of the tested products is 10 ~ 5, 10'7 and 10"9 M. The boxes are then kept in the incubator for 21 days, on the 21st day, the boxes are treated by the deposit in the upper layer of 100 μl of a solution of MTT (3- (5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolinium bromide) at 1 mg / ml prepared with RPMI 1640 medium for 3 hours at 37 ° C. After this period of time, the cell colonies are fixed by the addition of 2 ml of formalin per box.After the fixation for 24 hours, the formalin is evaporated and a number of cell colonies are colored, so both consists of metabolically active cells and whose surface area is greater than 100 μm2 is determined with the help of an inverted microscope.The average number of clonogenic cell clones determined by each experimental condition studied is expressed as a percentage relative to the average number of clones clone cell phones nicos counted under the condition of control and who have as equal to 100%. These values, expressed as the percentage relative to the control condition, are presented in Table I.

The results summarized in this table represent the mean values + _ standard deviation of the mean (SEM) established in 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 can: recruit clonogenic cells into the tumor (HT-29 cell lines at concentrations of 10 -5 M and 10 ~ 7, and MTX at concentrations of 10"7 M and 10" 9 M), this is said, induces a significant increase in the number of colonies of these cells compared to those obtained under the control condition, and then it makes them more sensitive to conventional treatment with cytotoxic agents, or they are able to directly inhibit the proliferation of these clonogenic cells (cell line MCF7 at the concentrations of 10"5 M and 10" 7 M). 2- Cytotoxic activity at the level of non-clonogenic cells: "MTT test" The influence of the compounds of formula (I) on non-clonogenic cells was evaluated with the help of the MTT colorimetric test. The principle of the MTT test is based on mitochondrial reduction by the methabolically active living cells of the MTT product (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide), which is yellow in color. color, to a product which is blue in color, formazan. The amount of formazan thus obtained is directly proportional to the amount «At > of living cells present in the culture wells. This amount of formazan is measured by spectrophotometry. The cell lines are maintained in the monolayer culture at 37 ° C in closed stopper culture discs containing basal medium MEM 25 HEPES (Minimum Essential Medium). This medium is completely suitable for the growth of a range of varied mammalian primary or diploid cells. This medium is then supplemented: with a 5% amount of decomplemented SVF (Fetal Bovine Serum) at 56 ° C for 1 hour, with 0.6 mg / ml L-glutamine, with 200 IU / ml penicillin, - with 200 μg / ml of streptomycin, with 0.1 mgm / ml of gentamicin.

The 12 human cancer cell lines which are used were obtained from the Ameri ca n Type Cul t ure Col l ec ti on (ATCC, Rockville, MD, USA). These 12 cell lines are: - U-373MG (ATCC code: HTB-17) and U-87MG (ATCC code: HBT-14), which are two gl ioblastomas, - SW1088 (ATCC code: HBT-12), which is an astrocytoma, A549 (ATCC code: CCL-185) and A-427 (ATCC code: HBT-53) which are two non-small cell lung cancers, HCT-15 (ATCC code: CCL-225) and LoVo (ATCC code: CCL-229) which two cancers color, T-47D (ATCC code: HBT-133) and MCF7 (ATCC code: HBT-22) which are two breast cancers, J82 (code ATCC: HBT-1) and T24 (code ATCC: HBT-4) which are two cancers of the bladder, PC-3 (code ATCC: CRL-1435 ) which is a prostate cancer.

From the experimental point of view, 100 μl of a cell suspension containing 20,000 to 50,000 (in accordance with the cell type used) cells / ml of the culture medium, were inoculated into 96 well multi-well plates of flat bottom and are incubated at 37 ° C, under an atmosphere comprising 5% C02 and 70% humidity. After 24 hours of incubation, the culture medium is replaced with 100 μl of the fresh medium containing either the various compounds to be tested at concentrations ranging from 10 ~ 5 to 10"10 M, or the solvent which serves to the dissolution of the products to be tested (control condition).

After 72 hours of incubation under the preceding conditions, the culture medium is replaced with 100 μl of a yellow solution of MTT dissolved in an amount of 1 mg / ml in RPMI 1640. The microplates are incubated for 3 hours at 37 ° C. and then centrifuged for 10 minutes at 400 g. The yellowish solution of MMT was removed and the blue formazan crystals formed in the cell are dissolved in 100 μl of DMSO. The microplates are then placed under agitation for 5 minutes. The intensity of the resulting blue color, and therefore of the conversion of the yellow MTT product to blue formazan by the still alive cells at the end of the experiment, is quantified by spectrophotometry with the help of a DYNATECH IMMUNOASSAY SYSTEM type apparatus at the wavelengths of 570 nm and 630 nm corresponding to the wavelength by maximum absorption of formazan and to the previous nozzle respectively. A software integrated in the spectrophotometer calculates the mean optical density values as well as the standard deviation (Est. Deviation) and the standard error of the values of the mean (SEM). By means of the non-limiting example, the results of the average optical density, expressed as a percentage relative to the average optical density measured under the control condition (equal to 100%), obtained with an isoflavonoid: genistein, in the 5 cell lines of tumor U-87MG, J82, HCT-15, T-47D and A549, will be given in Table II.

TABLE 2 xx + yy = mean value + standard error of the average control condition = 100% - (NS: p> 0.05; *: p <0.005; **; p <0.01; ***: p <0.001 ). 5 Genistein has a low antitumor power. This non-toxic product induces, when this is the case, the inhibition of the total cellular 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 effects marginal can be demonstrated. 3. Determination of maximum tolerated dose 15 (MTD): The evaluation of maximum tolerated doses was carried out in mice 4 to 6 weeks of age B6D2Fl / Jico. The compounds were administered intraperitoneally in increased doses that several from 2.5 to 160 mg / g. The MTD value (expressed in mg / kg) is determined from the observation of the survival rate of the animals during a period of 14 days after a single administration of the product under consideration. The variation of the weight of the animals is also monitored on this period. When the BAT value is greater than 160 mg / kg, the BAT value is considered as 160 mg / kg by defáult. Genistein is by defáult associated with a BAT equal to 160 mg / kg. This result suggests that the products of the family of isoflavonoids do not present any direct toxicity and can be used in high tissue concentrations, and therefore in high dosages. 4. Antiviral activity in vivo in combination with cytotoxic agents The assays were carried out in the models of: murine adenocarcinoma mammary hormone-sensitive MXT (HS-MXT), - lymphoma P 388, in the presence or in other modes of cytotoxic agents such as cyclophosphamia, etoposide, doxorubicin or vincristine. When the BAT value for a product was determined, its anti-tumor activity in vi was characterized at the doses of MTD / 2, MTD / 4 and MTD / 8 in the mammary adenocarcinoma model of murine origin HS-MXT and in the lymphoma model P388). This is the dose which presented the best antitumor activity in these different models which were selected and used in the context of the combined treatments with the cytotoxic agents. In all the examples presented below, any model (adenocarcinoma mammary HS-MXT or Línfoma 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 for dissolving the different compounds of formula (I) used is administered for 5 consecutive weeks and at the rate of 5 administrations (Monday, Tuesday, Wednesday, Thursday and Friday) per week. The following were determined using these tests: i) - survival rate of the mice This survival ratio was calculated in the form of a T / C ratio (Number of days (medium-sized mice (number of surviving treated) mice those of the mice which died medium-sized during the days groups of mice that preceded treated) those for the treated medium mouse) T = (Number of mice that died on the same day as the treated medium mouse) (Number of days (medium mice (Number of surviving mice treated the mice which died from medians during the days groups of mice that preceded treated ones) those for the medium control mouse) C - (Number of mice that died the same day as the control medium mouse This proportion represents the average survival time for the medium mouse of the treated mouse group, relative to the average survival time for the mouse medium of the mouse group of In addition, a molecule induces a significant increase (P < 0.05) in the survival of the animals when the proportion of T / C exceeds 130%. Otherwise, a toxic effect occurs when this T / C value is less than 70%. ii) - tumor growth, by measurement, twice a week (Monday and Friday), the surface area of transplanted tumors HS-MXT and P388. This surface area is calculated by taking the product from the value of the two largest perpendicular axes of the tumor. The value of these axes is measured with the help of a sliding caliper. 4. 1 Murine mammary adenocarcinoma (HS-MXT) The mammary adenocarcinoma model of murine MXT, which is the B6D2Fl / Jico mouse from 4 to 6 weeks old transplanted (HS-MX) sensitive to the i ....

Hormone is a model derived from the galactoform 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 example.

Treatment 1 Genistein was administered alone. The first injection of the product was carried out on the seventh day post-transplant (D /) for four consecutive 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 first injection of the product was carried out on the fourteenth post-transplant day (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. '- - •' - and > .; "i -. r, X, ryj '... .. and». **.. -. J -. .i. and ».

Treatment 3 Vincristine (VCR) was administered alone. The first injection of the product was carried out on the fourteenth post-transplant day (D14) for three consecutive weeks at the rate of 3 injections per week (Monday, Wednesday and Friday) and at the dose of 0.63 mg / kg.

Treatment 4 The etoposide (ETO) was administered alone. The first injection of the product was carried out on the fourteenth post-transplant day (D14) for three weeks 15 consecutive to the proportion of 3 injections per week (Monday, Wednesday, and Friday) and at the dose of 10 mg / kg.

Treatment 5 Genistein is co-administered with cyclophosphamide. In this case, the first injection of genistein was carried out on day seven post-transplant (D7) for four consecutive weeks at the rate of 5 injections per week (Monday, ^^^^ ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ^^^^^ ^^^^ ^ ^^^^^^ ^ ^^^^^^^ 8 ^^^^^^^^^^^^^^ ^ ^ ^ ^ ^ ^ ^ Tuesday, Wednesday, Thursday and Friday) at the dose of 20 mg / kg and the first injection of cyclophosphamide was carried out on the fourteenth post-transplant day (D14) for three consecutive weeks at the rate of three injections per week (Monday, Wednesday and Friday) at a dose of 10 mg / kg.

Treatment 6 Genistein is coadministered with vincristine. In this case, the first injection of genistein is carried out on day seven post-transplant (D79 for four consecutive weeks at the rate of 5 injections per week (Monday, Tuesday, Wednesday, Thursday and Friday) at a dose of 20 15 mg / kg and the first injection of vincristine was carried out on the fourteenth post-transplant day (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 20 Treatment 7 Genistein is co-administered with etoposide. In this case, the first injection of genistein is carried out on the seventh day after ^^^^^^^^^^^^^^^^^^^^^^^^^^ MS ^^^^^^^^^^^^^^^^^^^ ¿^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ) 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 etoposide was carried out on the fourteenth post-transplant day ( D14) for three consecutive weeks at the rate of three injections per week (Monday, Wednesday and Friday) at a dose of 10 mg / kg.

The results obtained for the survival period (Table III) by genistein will be given below.

TABLE III These results show that the co-administration of genistein with the cytotoxic agents: cyclophosphamides, vincristine or etoposide, significantly increase the mean survival time for the medium mouse of the different groups of mice treated, compared with the average survival time for the average mouse. of the control mouse group. further, this increase in the mean survival time for the medium mouse of the different groups of mice treated with these co-administrations is significantly greater than that obtained with the treatments involving geniethein or these cytotoxic agents used alone. The study of tumor growth, however, did not show the following results. In Table IV below, decreases (-) or increases (+) in the surface area of HS-MXT tumors induced by different treatments 1, 2, 3, 4, 5, 6 and 7 compared to the control condition on day 28 after tumor transplantation, that is, after 15 administrations of genistein and 6 administrations of the different cytotoxic agents used or otherwise in co-administrations with genistein. At day 28 post-transplant, 89% of control animals are still alive (this is 8 animals out of 9).

TABLE 4 These results show that the co-administration of genistein with the cytotoxic agents: vincristine and etoposide, significantly reduces a decrease in the growth of HS-MXT tumors which are greater than those induced by the treatments that involve genistein alone (which has no relevant clinical effect) or the last two cytotoxic agents used alone. 4.2 Lymphoma P 388: CDF1 mice from 4 to 6 weeks of age received a transplant consisting of a piece of the P388 tumor (obtained from a laboratory tumors maintained in the laboratory) subcutaneously Gc ^^^^^ «g * ^^ ~ on the right side to the day DO. To be in a situation similar to the clinical reality, we wait for day 5 post-transplant (D5) before starting the treatment. This was because after this period of time, P388 subcutaneous tumors are palpable. By means of the example, the results obtained with genistein alone or in combination with vincristine are reported at low.

Treatment 1 Genistein is administered alone, the first injection of the product is carried out on the fifth day post-transplant (D5) at the rate of 5 injections per week (Monday, Tuesday, Wednesday, Thursday and Friday) for five consecutive weeks and a dose of 40 mg / kg.

Treatment 2 Vincristine (VCR) was administered alone. The first injection of the product is carried out on day five post-transplant (D5) at the rate of 3 injections per week (Monday, Wednesday and Friday) for three consecutive weeks and at a dose of 0.63 mg / kg.

Treatment 3 Genistein is co-administered with vincristine. In this case, the first injection of genistein was carried out at day five post-transplant (D5) at the rate of 5 injections per week (Monday, Tuesday, Wednesday, Thursday and Friday) for five consecutive days at the dose of 40 mg / kg and the first injection of vincristine was carried out at day five post-transplant (D5) at the rate of 3 injections per week (Monday, Wednesday and Friday) for three consecutive weeks at the dose of 0.63 mg / kg . The results obtained with treatments 1, 2 and 3 in the survival times for the mice are presented in Table 5 below.

TABLE 5 These results show that co-administration of genistein with vincristine ...-, - M ^ a ^ m significantly increase the average survival time for the medium mouse of the different groups of mice treated in this way compared to the average survival time for the medium mouse of the group of control mouse. Furthermore, this increase in the mean survival time for the medium mouse of the different groups of mice treated in this way is highly significant compared to the average survival time by the medium mouse of the different groups of mice treated with genistein or vincristine which are used alone Examples of the method of using the compounds of formula I in mono or polychemotherapy protocols with cytotoxic agents will be given below.

A. Solid tumors 1 / Lung cancers 1.1 Non-small cell type (advanced status): - for the recommended protocol (T. Le Chevalier et al., J. Clin. Oncol. 1994; 12. 360-367), be added Intravenous infusions of genistein or other isoflavonoids: this cure was repeated 8 times 1. 2 Small cell type (advanced state): - for the recommended CAV or VAC protocol (B.J. Roth et al., J. Clin. Oncol. 1992; 10: 282-291), the infusions of iso flavonoids are added: . -MíS? Fy.'A this cure was repeated 6 times every 21 days. - for the recommended protocol Pt-E (B.J. Roth et al., J. Clin. Oncol., 1992; 10: 282-291) the infusions of genistein were added each cycle is repeated every 21 days and the cure 15 comprises six cycles. 1. 3. Bronchial cancer without locally advanced or metastatic small cells: 20 • monochemotherapy: It is possible for the cure to understand the repetition of the 4-week cycle. • combination genci tabina / cisplatino: the cure includes the repetition of this cycle every 21 days. 2 / Breast cancers CMF protocol as adjuvant treatment for operable breast cancer (G: Bonnadonna et al., N. Engl. J. Med., 1976; 294: 405-410): each cycle was repeated every 28 days and the cure comprises 6 cycles. - AC Protocol (B. Fisher et al., J. Clon Oncol, 1990; 8: 1483-1496) as adjuvant treatment: each cycle is repeated every 21 days and the cure includes 4 cycles - Breast cancers with metastases: in the FAC protocol (AU Buzdar et al., Cancer 1981; 47: 2537-2542) and its different adaptations, isoflavonoid infusions are add in accordance with the following scheme (not limiting): each cycle is repeated every 3 weeks until a new progression of the disease is diagnosed. - in each CAF protocol (G. Falkson et al., Cancer 1985; 56: 219-224): , *? ^^^^^^^^ & ^^^^^ sj = l? f & each cycle was repeated every 28 days until a new progression of the disease was diagnosed: 10 - in the CMF protocol: fifteen this cycle is repeated every 3 to 5 weeks and the cure includes 6 cycles: - in the CMF-VP protocol: This cure is repeated every 4 weeks. - in the FEC protocol: This cure is repeated every 3 weeks. - in the MMC-VBC protocol (C. Brambilla et al., Tumori, 1989, 75: 141-144): This cure is repeated every 28 days until the progress of the condition is diagnosed. - in the protocol (S. E. Jones et al., J. Clin. Oncol., 1991; 9: 1736-1739): the cure comprises two cycles of 21 days apart and then requires evaluation. The infusions of isof lavonoides can also be combined with the treatment of cancers ^ > , # of breast with metastases when the toxoid is used, for example: with paclixatel (FA Holmes et al., J. Natl Cancer Inst. 1991; 83: 1797-1805) in the treatment of forms with metastases which can be resistant to the anthracyclines: This cycle is repeated every 21 days until a new progression of the disease is diagnosed. with docetaxel (CA Hudis et al., J Clin. Oncol., 1996; 14: 58-65), in locally advanced or metastatic breast cancer, resistant or relapsing after cytotoxic chemotherapy (comprising an anthracycline) or in relapses during an adjuvant treatment: i »° ^ ^ at - ^^ -« - This cycle was repeated every 21 days for a cure of 2 cycles or until a progress of the condition appears. in dose escalation protocols that combine a transplantation of medullarly autologous cells and cells of the peripheral bloodstream as a consolidation of the first-line treatment, for example: CPB protocol (W. P. Peters et al., J Clin. Oncol., 1993; 11: 132-1143), in which the i.v infusion of the derived cells takes place on days D-x, Do and Dx: - CTCb protocol (K. Antman et al., J. Clin. Oncol. 1992; 10: 102-110), in which the i.v. of the derived cells take place on day D0: CTM protocol (L. E. Damcn et al., J. Clin Oncol 1989, 7: 560-571 and I-C-Henderson et al., J. Cellular Biochem. 1994 (Suppl 18B): 95) in the * iL? i i t i. i which the i.v infusion of hematopoietic derived cells takes place in D0: 3 / Gynecological Cancers 3.1 Ovarian cancer: - for the treatment in particular of metastatic ovarian carcinomas i) PAC protocol (GA Omura et al., J .. Clin. Oncol. 1989; 7: 457-465): infusions of Isoflavonoids were administered according to the following scheme: this cycle is repeated every 21 to 28 days and the cure comprises 8 cycles. ii) altretamine protocol, in accordance with A. Marietta et al. (Gynecol Oncol 1990; 36: 93-96): the cure comprises two cycles, days apart. ii) paclitaxel protocol: the isoflavonoids can be added to the paclitaxel protocol as described by W.P. McGuire et al. (Ann, Intern. Med. 1989; 111: 273-279): the cure comprises two of these cycles, 28 days apart (with the evaluation at the end). - for the treatment of metastatic and refractory ovarian carcinomas, isoflavonoids can be added to the second line protocol, based on the topotecan: the cure comprises two cycles, 21 days apart (with evaluation at the end) in accordance with A. P. Kuldelka et al. (J_ Clin Oncol 1996: 14: 1552-1557). j »& & & amp; & amp; 3.2 Trophoblastic tumors: in low risk patients, isoflavonoids can be combined with the protocol described by H. Takamizawa et al. (Semin. Surg. Oncol. 1987; 3: 36-44): (MTX-DACT protocol). 3. 3 Uterine cancers: isoflavonoids can also be combined with the CAV (or VAC) protocol in accordance with the scheme below: the cure involves a repetition of this cycle every 21 days. - in the FAP protocol: the cure includes the repetition of this cycle every 21 or 28 days. 4 / Testicular and prostate cancers Isoflavonoids can also be combined with testicular cancer protocols: the cure comprises three cycles, at the rate of one cycle every 21 days.

/ Cancers of the bladder 5 - the isof lavonoids can be combined with the CISCA2 protocol (also called PAC) The cycle has to be repeated every 3 weeks. - in the MVAC protocol (in accordance with CN Sternberg et al., J. Urol. 1988; 139: 461-469): twenty ^ ¡^^^^^^^^^^^ this cycle is repeated every 4 to 5 weeks, at least for 2 cycles. 6 / Naso-arginine carcinomas / head and neck cancers Isoflavonoids can be legitimately combined with the polychemotherapy protocols used in the treatment of these cancers: 6. 1 Nasopharyngeal Cancers: - Protocol ABVD: the cure comprises 1 to 6 repeated cycles at the rate of 1 cycle every 4 weeks. ? ^^ gg ^? ß 6.2 Head and neck cancers with metastases: - in the Pt-FU protocol (for example: for cancers of the pharynx): in accordance with the DVAL Study Group (New Engl JM 1991; 324 : 1685-1690): The cure comprises two cycles, the proportion of 1 cycle every 3 weeks. 7 / Carcinomas of soft tissue Isoflavonoids can be introduced into a protocol such as the CYVADIC protocol: in accordance with H.M. Pinedo et al. (Cancer 1984; 53: 1825): the cure includes the repetition of this cycle every 4 weeks, first for 2 cycles. 8 / Prostate cancer refractory to the hormone, with metastases - in the VBL-estramustine, in accordance with G.R. Hudis et al. (J. Clin. Oncol. 1992; 10: 1754-1761): a last cycle of treatment for 6 weeks and followed by 2 weeks of free interval. 9 / Embryonic cell cancers i) for tumors with a favorable prognosis: Pt-E protocol, in accordance with G.J Bosl et al. (J. Clin. Oncol. 1988: 6: 1231-10328) fifteen the cure comprises 4 cycles, at the rate of 1 cycle every 21 or 28 days. Ii) for tumors with metastases: PEB protocol, in accordance with S.D. Williams et al. (N. Eng. J. Med. 1987; 316: 1435-1440): 25 ~ .Jnyi? Yes? ~ the cure comprises 4 cycles, at the rate of 1 cycle every 21 days.

/ Renal cancers metastatic renal carcinoma: isoflavonoids can be introduced into the protocol described by M. J. Wiikinson et al. (Cancer 1993; 71: 3601-3604): the cure comprises two cycles of 28 days apart. Nephroblastoma: I soflavonoids can be introduced in the DAVE protocol: at the rate of one cycle every 3 to 4 weeks 11 / Cancers of the digestive tract 11.1 Cancers of the esophagus: - isoflavonoids can be introduced into the FAP protocol in accordance with: this cycle is repeated every 3 to 4 weeks i i i? ^ ¿^. : 11.2 Stomach cancers in advanced gastric carcinomas and / or with metastases: EAP protocol (in accordance with P. Preusser et al., J Clin. Oncol. 1989; 7: 1310): at a rate of 1 cycle every 28 days. - FAMtx protocol: in accordance with J.A, Wils et al. (J. Clin. Oncol., 1991; 89; 827): the cure comprises two cycles, 28 days apart , S-agjf - in certain patients, the protocol or its variant (epirubicin, which replaces doxorubicin) can be used in accordance with the following scheme: 12 / Colorectal cancers - isoflavonoids can be introduced into the protocol by the FU-Levamisole adjuvant treatment of colorectal cancer (in accordance with CG Moertel et al., N. Eng. J. Med. 1990; 322: 352): the treatment in the form of a bolus with 5-FU being repeated every week after the induction phase of D1-D5, for two 52 weeks; that with an isoflavonoid is repeated at the same proportion, the day of the bolus 5-FU and then the next 2 days. for the treatment of colorectal cancer which is refractory to treatment with 5-fluorouracil (5-FU) and with metastases: in accordance with M. L. Rothenberg et al. (J. Clin. Oncol., 1996; 14: 1128-1135): the cure comprises two cycles, 42 days apart 13 / Kaposi sarcomas isoflavonoids can be combined with the two protocols using anthracyclines formulated in the form of liposomes: i) protocol described by P. S. Guill et al. (J. Clin Oncol 1995, 13: 996-1003) and C. A. Presant et al. (Lancet 1993; 341: 1242-1243): the cure comprises two repeated cycles at an interval of 28 days before the evaluation of the effects. ii) protocol of M. Harrison et al. (J Clin Oncol 1995, 13: 914-920): the cure comprises two repeated cycles at an interval of 28 days before the evaluation of the effects. 14 / Metastatic melanomas Isoflavonoids can also be incorporated into the combined protocols to treat malignant metastatic melanomas: DTIC / TAM protocol: in accordance with G. Cocconi et al. (N. Eng. J. Med. 1992; 327: 516), the cure comprises the repetition of 4 cycles, at the rate of 1 cycle every 21 days, in accordance with the following scheme: the cure comprises 4 cycles at the rate 1 cycle every 21 days / Neuroendocrine carcinoma isoflavonoids can be combined with the protocol described by C. G. Moertel et al. (Cancer 1991; 68: 227): Pt-E protocol: the cure comprises two repeated cycles every day 16 / Pancreatic cancer advanced pancreatic adenocarcinoma: the isoflavonoids can be combined with the treatment with gencitabine according to the protocol of M. Moore et al. (Proc. Am. Soc. Clin. Oncol. 1995; 14: 473): B. Oncohematology 1 / Acute leukemia of adults 1.1. acute lymphoblastic leukemia 1.1.1 linker protocol Isoflavonoids can be added to the linker-induction protocols of chemotherapy and consolidation chemotherapy (see CA Linker et al., 1987, 69: 1242-1248 and CA Linkeer et al., 1991; 78: 2814-2822) in accordance with the following schemes: 1) induction of chemotherapy: iii) consolidation chemotherapy (regimen A): The consolidation cure A comprises 4 consecutive cycles as those described above = 1, 3, 5 and 7 cycles. iii) consolidation chemotherapy 15 (regimens B and C): The regimens described below correspond to consolidation cycles 2, 4, 6 and 8 (regimen B) and 9 (regimen C), described by C.A Linker et al .: twenty . ,, j. ^ * J2ia ^ * ^ lí -, 1. 1.2 Hoelzer Protocol The claimed products can 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) in accordance with the following scheme: i) induction of chemotherapy / phase 1: • g ^^^^^ & feg ^^^^^^ jk iii) reinduction / phase 1 chemotherapy fifteen iv) reinduction chemotherapy / phase 2: 20 ¿^ -m ^^ yfc ^^^ b¡¡¡ ^^^^ 1.2 Acute myeloid leukemias: 1.2.1 Treatment of adults of any age Isoflavonoids can be added, in accordance with the scheme below, to the treatment incorporating the standard dose of cytarabine previously described by RO Dilleman et al. (Blood, 1991; 78: 2520-2526), Z. A. Arlin et al. (leukemia 1990; 4: 177-183) and P.H. Wiernik et al. (Blood 1992; 79: 313-319): 1. 2.2 Adult treatment under 60 years of age i) induction of chemotherapy: This cycle of induction incorporates the administration of cytarabine in a high dose in accordance with the following scheme: fifteen (to reduce the risk of S.N.S. toxicity, in the event of renal failure, adjust the dosage to the creatinine evaluation) in accordance with L. E. Damon et al. (Leukemia 1994; 8: 535-541), G. L: Phillips et al. and S &i? ix andYy & & amp; (Blood 1991; 77: 1429-1435) and G. Smith et al. (J. Clin. Oncol. 1997; 15: 833-839). 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 RJ Mayer et al., N. Engl. J. Med. 1994; 331: 896-903): iii) consolidation chemotherapy (with a high dose of cytarabine): The cycle, described below, would have to be repeated twice and adapted in accordance with G. L Phillips et al, (Blood 1991; 77: 1429-1435); S.N. Wolff et al, (J. Clin. Oncol., 1989; 7: 1260-1267); -, "& .. *, I fc .M-safeS-taS *.

R.J. Mayer et al. (N. Engl J. Med. 1994; 331: 896-903): 1. 2.3 Treatment of adults aged 60 years or older The claimed substances can be added to the consolidation chemotherapy protocols below: i) in accordance with 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 1992; 79: 313-319): _ .. .. í 10 ii) according to R. J, Mayer et al (N. Engl. J. Med, 194; 331: 896-903): 15 twenty "& iéÉ¿É¡¡?? & £ aí g¿ lg ^ g ** j && amp; amp; iii) in accordance with 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): This protocol comprises an autologous bone marrow transplant (performed on day D0): or iv) in the case of HLA-compatible allogeneic bone marrow transplantation in accordance with: P. J. Tutscha et al. Blood 1987; 70: 1382-1388, F.R. Applebaum et al., Ann. Int. Med. 1984; 101: 581-588: 2 / Chronic adult leukemias 2.1 Chronic myeloid leukemia In the myeloblastic phase, the isoflavonoids can be added to the HU-Mith treatment, described by C.A. Koller et al. (N. Engl. J. Med. 1986; 315: 1433-1438): 2. 2 Chronic lymphocytic leukemia 2.2.1 FCG-CLL protocol Isoflavonoids can be added to combinations of "chlorambucil puised" as described by E. Kimby et al. (Leuk, Lymphoma 1991; 5 (Suppl.) 93-96) and by FCGCLL (Blood 1990; 75: 1422-1425): 2. 2.2 Fludarabine-CdA protocol in accordance with H.G. Chun et al. (J, Clin. Oncol. 1991; 9: 175-188), M. J. Keating et al.

(Blood 1989; 74: 19-25 / J Clin Oncol 1991; 9: 44-49) and A. Saven et al. (J Clin Oncol 1995, 13: 570-574): 3 / Lymphoproliferative disorders 3.1 Hodgkin's disease Isoflavonoids can be incorporated into polychemotherapy protocols conventionally used for the treatment of Hodgkin's lymphoma: 3. 1.1 ADVB protocol in accordance with G. Bonnadonna et al (Cancer Clin. Triáis 1979; 2: 217-226) and G. P.

Canellos et al. (N. Engl. J. Med. 1993; 327: 1478-1484): the cure comprises 6 cycles, the proportion of 1 cycle every 28 days 3. 1.2 MOPP / ABDV protocol in accordance with G. Bonnadonna et al. (An. Intern. Med. 1986; 104: 739-746) and G.P. Canellos et al. (N. Engl. J. Med. 1993; 327: 1478-1484): The MOPP protocol must alternate with the ABVD protocol (cf. &3.1.1) every 28 days and the cure comprises 6 cycles: awfe% a - »&, 3. 1.3 Stanford Protocol V in accordance with N. L. Barlett et al. (J, Clin Oncol, 1995; 13: 1080-1088): the cure comprises 3 cycles, at the rate of 1 cycle every 28 days. 3. 1.4 EVA protocol in accordance with G. 0. Canellos et al. (Proc. Am. Soc. Clin. Oncol. 1991; 10: 273): the cure comprises 6 cycles, at the rate of 1 cycle every 28 days. 3. 1.5 Protocol B-CAVe in accordance with W.G. Harker et al. (Ann Intern. Med. 1984; 101: 440-446): the cure comprises 8 cycles, at the rate of 1 cycle every 28 days. 3. 2 Non-Hodgkin lymphomas 3.2.1 of low degree of malignancy i) CVP protocol in accordance with C.M. Bagley et al. (Ann Intern Med 1972; 76: 227-234 and C.S. Potlock et al. (Blod 1976; 47: 747-756) This cycle is repeated every 21 days until the maximum response ii) Protocol I-COPA - in accordance with RV Smalley et al. (N.

Eng. J. Med. 1992; 327: 1336-1341) The cure comprises 8 to 10 cycles, the proportion of one cycle every 28 days. iii) Fludarabine-CdA protocol - in accordance with P. Solol-Celigny et al. (Blood 1994; 84 (Supp.1): 383 a), H. Hoeschester et al .; (Blood 1994; 84 (Supp.1): 564a and A.C. Kay (J. Clin. Oncol. 1992; 10: 371-377).

For fludaribine, each cycle is repeated every 28 days, for cladribine, each cycle repeating every 35 days. 3. 2.2 degree of intermediate malignancy i) CHOP or CNOP protocol - in accordance with EM McKelvey et al. (Cancer 1976; 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) For the CHOP protocol Mitoxantrone (N) can be used to replace (CNOP protocol) doxorubicin in patients older than 60 (dose: 12 mg / m2 as bolus i.v. per day Di of each cycle). 15 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 - in accordance with P. Klimo et al. (Ann, Intern Med 1985, 102: 596-602) and I.. Cooper et al. (J. Clin. Oncol., 1994; 12: 769-778) 25 This treatment protocol extends over 12 weeks and corresponds to 1 cycle. iii) VACOP-B protocol - in accordance with J. M. Connors et al. (Proc. Am. Soc. Clin. Oncol. 1990; 9: 254): ^^^ ¿& j ^ k¡ £ # £ ijl? ^ £ ^^^ 10 fifteen each cycle remains for 12 weeks. ív) - Protocol m-BACOD / M-MACOD 20 - in accordance with M.A. Shipp et al. (Ann Int. Med. 1986; 140: 757-765) and T. Skann et al. (J. Clin. Oncol. 1983; 1: 91-98) The cure comprises 10 cycles, at the rate of 1 cycle every 21 days. iv) ProMACE / CytaBOM protocol - in accordance with D. L. Longo et al (J. Clin. Oncol., 1991; 9: 25-38): The cure comprises 6 cycles, the proportion of one cycle every 14 days 3. 2.3 degree of malignancy low or intermediate i) ESHAP rescue protocol - in case of relapse or in case of failure of the first line of treatment, in accordance with W.S. Velasquez et al. (J. Clin. Oncol. 1994; 12: 1169-1176) The cure comprises 6 cycles, at the rate of 1 cycle every 28 days. ii) MINE rescue protocol - in case of relapse or in the case of failure of the first line of treatment, in accordance with F. Cabanillas et al. (Semin Oncol 1990, 17 (Supp 10) 28-33) This cycle is repeated every 21 days 3 Non-Hodgkin lymphomas: Burkitt lymphoma, small cell lymphoma, lymphoblastic lymphoma 3. 1 Magrath Protocol The claimed products can be combined with the Magrath protocols in accordance with the following schemes: i) cycle 1 - in accordance with I.T. Magrath et al. (Blood 1984; 63: 1102-1111) ii) cycles 2 to 15 in accordance with I.T. Magrath et al (1984) also fifteen twenty The cure comprises 14 cycles, at a rate of 25 of a cycle every 28 days. 3. 4 Macroglobulinemia aldenstrom 3.4.1 CVP protocol in accordance with 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): the cure is continued indefinitely (1 cycle every 21 days). 3. 4.2 Fludarabine-CdA Protocol - in accordance with H.M. Kantarjian et al (Blood 1990; 75: 1928-1931) and M.A. Dinopoulous et al (Ann, Intern. Med. 1993; 118: 195-198): - & k fc # - ^ o the cure comprises 6 to 12 cycles of 28 days apart in the case of fludarabine and 2 cycles of 28 days apart also in the case of cladribine. 3. 5 Multiple myeloma 3.5.1 MP protocol in accordance with R. Alexanian et al.

(JAMMA 1969: 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): .fr • - - »* ~«.,. the cure comprises at least 12 cycles, at the rate of 1 cycle every 4 to 6 weeks. 3. 5.2 VAD protocol in accordance with B. Barlogie et al, (N. Engl. J. Med. 1984; 310: 1353-1356): 3. 5.3 Protocol MP-a interferon in accordance with O. Osterborg et al [Blood 1993; 81: 1428-1434): the cure includes the indefinite repetition of this cycle, at the rate of 1 cycle every 42 days. 3. 5.4 VCAP or VBAP protocol in accordance with S.E. Salmon et al (J Clin Oncol 1983: 1: 453-461): VCAP protocol: VBAP protocol: cyclophosphamide is replaced with carmustine (BCNU), the remainder is identical: C. INFANT TUMORS - Pediatric Oncology Isoflavonoids can also be incorporated into the chemotherapy protocols for the treatment of pediatric tumors in order to increase antitumor efficacy while reducing the severity of side effects through the action of the patient. recruitment and mobilization of clonogenic cells and the possibility of reducing active doses. 1 / Ewing's sarcoma / primitive neuroectodermal tumor Isoflavonoids can be introduced into the VCR-Doxo-CY-Ifos-Mesna-E (ED Bergert et al., J. Clin. Oncol. 1990; 8: 1514-1524; WH Meyer et al., J. Clin. Oncol. 1992; 10: 1737-1742): the cure comprises 6 to 10 of these cycles depending on the initial severity of the sarcoma and the extent of the response. 2 / Acute lymphoblastic leukemia of infants 2.1. Induction of chemotherapy (days D? -d3o.

Isoflavonoids can be added to the recommended protocols (PS Gaynon et al., J. Clin Oncol., 1993, 11, 2234-2242, J. Pullen et al., J. Clin. 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): ¡^^ Sg ^^ j ^^^ já ^ Jf c ^ depending on the result of the examination of the bone marrow, the step to the consolidation phase is done to day D28 of the treatment protocol. 2. 2 Maintenance maintenance / chemotherapy Isoflavonoids can be introduced into the maintenance protocol (PS Gaynon et al., J. Clin 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) in accordance with the following scheme: 3 / acute myeloid leukemia of infants The isoflavonoids are added to the induction and consolidation / maintenance protocols in accordance with the following schemes: 3. 1 Induction of chemotherapy In accordance with Y. Ravindranath et al., J. Clin. Oncol. 1991; 9: 572-580; ME. Nesbit et al., * * J J .. Clin. Oncol. 1994; 12: 127-135; RJ Wells et al., J. Clin. Oncol. 1994; 12: 2367-2377): this cycle repeats from the D 2 Í 3. 2 consolidation / maintenance chemotherapy In accordance with Y. Ravidranath et al., J.

Clin. Oncol. 1991; 9: 572-580; ME. Nesbit et al., J.

Clin. Oncol. 1994; 12: 127-135; R. J. Wells et al., J. Clin. Oncol. 1994; 12: 2367-2377): 4 / Infants' Hodgkin Disease Isoflavonoids can be added to the MOPP-ABVD protocol in accordance with EA Gehan et al. (Cancer 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): This cycle should be repeated 6 times at the rate of 1 cycle every 8 weeks, the cure includes 6 cycles. If an autologous bone marrow transplant (autograph) 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 _ £ _ * -? «AAi. .. and, J., i, i, i, RJ Jones et al (J. Clin. Oncol 1990, 8, 527-537) can be used in accordance with the following scheme (the place to date D0): / Lymphoblastic lymphoma of infants Flavonoids can also be combined with the chemotherapy induction protocols (AT Meadows et al., J. Clin Oncol 1989, 7: 92-99 - C. Patte et al., Med. Ped. Oncol. 1992; : 105-113 and A. Reiter et al., J. Clin. Oncol. 1995; 13: 359-372) and the maintenance of the chemotherapy protocols: . «J-aj jfe? ..- r. , i »5 &a-». . 1 Induction chemotherapy . 2 Maintenance chemotherapy according to the following scheme the cure comprises 10 cycles 6 / Pediatric Neuroblastoma The recommended Doxo-E-Cy-Pt protocol for polychemotherapy is adapted from R. P. Castleberry et al. (J. Clin Oncol 1992, 10: 1299-1304), A. Garavanta et al. (J. Clin Oncol, 1993; 11: 1770-1779) and D. C. West et al. (J. Clin. Oncol. 1992; 11: 84-90): The evaluation of the therapeutic response is made after 9 weeks to decide on the attitude: surgical resection, radiotherapy or new chemotherapy. 7 / Pediatric Osteosarcoma The isoflavonoids can be added to the 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): í. - 10 8 / Rhabdomyosarcoma of infants 15 The Vcr-Dact-CY-Mesna protocol (H. Maurer et al., Cancer 1993; 71: 1904-1922 and LR Mandell et al., Oncology 1993; 7: 71-83) may include infusion iv of isoflavonoids according to the following scheme: » Mt * mai * ÍSS &8d &aA? *? ~, And. iji ?. I.l i §- = te ^^ fe3for. 0. 015 mg / kg OV D1-D5, D22- dactinomycin (daily dose D27, maximum D43-D47: 0.5 mgl • cyclophos-2.2 g / m) as a 1. v, Di, D 22, D 43 f amide infusion of 1 hour • mesna 360 mg / m as a 1. v D 1, Dr 2, D 43 infusion of 1 hour every 3 hours for 5 doses At the end of the 9th. Treatment week, the effectiveness should be evaluated in order to decide the future course of action (surgery, radiotherapy, continuation of chemotherapy). 9 / Tumor of ilms in Infants 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): twenty This protocol is started after surgical resection. In case of autologous bone marrow transplantation (autograph) in accordance with A. Garaventar et al. (Med. Pediatr Oncol 1994; 22: 11-14), the E-Thio-Ci protocol can be modified as follows: The bone marrow transplant took place on day D0. It is noted that in relation to this date, the best method known by the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention. Having described the invention as above, the content of the following is claimed as property. ** > . «A» - í í? A ^ afe ^

Claims (8)

1. Composition which has an activity in the proliferation of clonogenic cells in tumors and which is characterized in that it comprises a therapeutically effective amount of an isoflavonoid or of a chromone type analogue.

2. Composition according to claim 1, wherein the isoflavonoid is selected from the compounds of formula: R » characterized in that: Ri, R2, R3 and R4, are independently selected, from each of the others, of H, OH, a C? -C alkoxy group, a -OCOR7 group, R7 being a C1-C4 alkyl group, at least one of the substituents Ri, R2, R3 or R4 are different from H and it is possible for R2 and R3 to together form a methylenedioxy group, Rs is selected from H, OH, an alcoholic group C? -C a group O- glycosyl and a cyclohexyl group, R6 is selected from a cyclohexyl group, a phenyl group and a phenyl group substituted 1 to 3 times with groups selected from H, OH and a C1-C alkoxy group and - denotes either a double bond , or a simple link.

3. - Composition according to claim 2, characterized in that the isoflavonoid is selected from genistein, daidzein and biocanin A.

4. Use of an isoflavonoid or a chromone-like analog for the manufacture of a medicament intended to interfere with the generation of clonogenic cells in tumors after treatment of these tumors by at least one cytotoxic agent.

5. the use of a compound selected from the compounds of the formula: ^ * > - in which in the formula: - Ri, 2 3 and R 4 - are independently selected, from each of the others, of H, OH, a C 1 -C 4 alkoxy group, a group -OCOR, R 7 being an alkyl group C? -C, at least one of the substituents Ri, R2, R3 or R4 are different from H and are possible -T :. ' i? For the R2 and R3 together form a methylenedioxy group, R5 is selected from H, OH, a C? -C4 alkoxy group, an O-glycosyl group and a cyclohexyl group, RD is selected from a group cyclohexyl, a phenyl group and a phenyl group substituted 1 to 3 times with groups selected from H, OH and a C 1 -C 4 alkoxy group, 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 tumors after treatment of these tumors by at least one cytotoxic agent.

6. Use according to claim 5 in which genistein, daidzein and biscanin A.

7. A method of chemotherapeutic treatment of a tumor in a patient by at least one cytotoxic agent, characterized in that it comprises the administration in the course of treatment of a cytotoxic agent of a therapeutically effective amount of an isoflavonoid or of a chromone-like analogue. 12Q

8. Method according to claim 7, characterized in that the isoflavonoid or analogue of the chromone type is administered at the start of the chemotherapeutic treatment at the beginning of each cycle of chemotherapeutic treatment.