ZA200100239B - Therapeutic composition based on flavonoids for use in the treatment of tumors with cytotoxic agents. - Google Patents

Description

Ya : i Nr Btf WO 00/03706 PCT/FR99/01714

E | 20010239 "compounds 6f flavonoid type in the treatment of cancers on : by cytotoxic agents.

Ba Eg i Acancer is a disorder of the somatic genes, in

BE © "the course of which genetic dysfunctions worsen as the - tumour progresses from the state of precancerous lesion

ERR that of malignant transformation, the cancerous tumour becoming metastatic and often resistant to cytotoxic medicines. i )

Despite the very considerable efforts conducted in all the developed 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, cancer is placed second, just after cardiovascular diseases, as the cause of mortality.

In terms of newly diagnosed cancers, the : + distribution between solid tumours and haematological (bone marrow, blood, lymphatic system) neoplasias shows that 9 cancers out of 10 are solid tumours. Unlike that which is’ observed in haematological oncology (therapeutic success in 40 to 90% of blood cell ‘cancers), only a small number .of advanced or TT . disseminated solid’ tumours respond to only : chemotherapeutic treatments. It is partly for this reason - that the overall mortality due to cancer : increased in the U.S.A. between 1973 and 1992. . It- is not, unfortunately, certain that this : tendency may be reversed only by the appearance, alongside the ‘established chemotherapeutic arsenal, of novel antitumour medicines such as taxanes (paclitaxel and docetaxel), which interfere with the formation of microtubules (W.P. McGuire et al., Am. Intern. Med., 1989), camptothecin-derived topoisomerase I inhibitors

“rn va 20010239 . ' - 2 = . (topotecan and irinotecan), vinorelbine (novel alkaloid derived from the periwinkle), gemcitabine (novel

Cytotoxic antimetabolic agent), raltitrexed i (thymidylate synthetase inhibitor) or miltefosine £ 5 (first: representative of the alkylphosphocholine oe or as a second choice, to the medicines for which the specific activity is now well recognized, . such as : doxoriibicin, cisplatin, vincristine, methotrexate, 5- fluorouracil. g

One of the most difficult current problems of anticancer chemotherapy is due to the fact that numerous populations of malignant -cells exhibit considerable resistance to established cytotoxic substances. Most commonly, this situation results from the existence of multiresistance genes or from the frequency of genetic mutations in certain types of tumour. Thus, the treatment of cancers requires novel approaches which are complementary to those currently implemented, and which - are intended to more successfully combat the extension and heterogeneity of - the tumour load, and the acquisition of “multi- ~ cytotoxic-drug” resistance.

Among these novel approaches, some are already promising. This is the case of the induction of apoptosis, and the inhibition of tumour angiogenesis and of metastatic processes, not to. mention gene therapy or immunotherapy. : oo } The inventors were interested in a different - 30 approach. The objective sought was to make the population of tumour cells more sensitive to reference : anticancer treatments, in order to achieve a double . ‘benefit: - 1) increase the cytotoxic activity and therefore the effectiveness, and 2) decrease the frequency and severity of certain side effects, due to the reduction in dosage which might follow the induction of the increase in the antitumour effectiveness.

cas Lo cUUTULOY ‘ It is this strategy which is behind the. discovery of an original mechanism caused by substances - which have weak antitumour power, or are lacking this power - but which are capable of inducing a very significant . increase in the cytotoxic activity of proven anticancer medicines. This original mechanism comes from the possibility for these substances either to stimulate the recruitment of clonogenic cells within the tumour, making it more sensitive to the conventional treatment with cytotoxic agents, or to inhibit the proliferation of clonogenic cells, thus contributing to the regression of the tumour.

A subject of the present invention is thus the oo use, in the treatment of cancers with at least one antitumour agent chosen from cytotoxic agents, of a compound having activity on the proliferation of clonogenic cells, chosen from flavonoids and in particular the compounds of formula:

R,

Re Oe . ( (LC ("

Rs Rs

R4 O this being a formula in which: - R,, R,, R, and R, are chosen, independently of each other, from H, OH, a C,-C, alkoxy group and an -OCOR, group, R, being a C,-C, alkyl group, at least one of the substituents R,, R,, R, or R, being other than H, and R, TT - 25 and R, possibly forming together a methylenedioxy group, : - Ry; is chosen from H, OH, a C,-C, alkoxy group and an

O-glycosyl group, - R, is chosen from a cyclohexyl group, a phenyl group : and a phenyl group substituted 1 to 3 times with groups ) 30 chosen from H, OH and a C,-C, alkoxy group, - and ...... designates either a double bond or a single bond. :

= | 20010239 1 eo. at @ [EI TAC

N The cytotoxic agents can be used at their ak conventional’ dose, and in this case, their effectiveness is improved, or at lower doses given the .

NE increase in their antitumour effectiveness. : E 1 LE 5 ~....A_subject of the present invention is also a

JU composition having “activity on the proliferation of e 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 amount of a flavonoid and in particular of a compound i of formula I chosen from the compounds of formula:

Ra © Re .

T LX 8

Ry Rs

Re ©O this being a formula in which: = Ri, R2;, R; and Ry; are chosen, independently of each other, from H, OH, a Ci-Cs alkoxy group and an -0OCOR, group, Ry being a C;-C, alkyl group, at least one of the ~ substituents R;, R,, Rj or Ry being other than H, and R; and R3 possibly forming together a methylenedioxy group, - Rs is chosen from H, OH, a Ci;-C4 alkoxy group and an

O-glycosyl group, - Rg is chosen from a cyclohexyl group, a phenyl group and a phenyl group substituted 1 to 3-times with groups TT © 25 chosen from H, OH and a Ci1—-Cyq alkoxy group, ’ - and TUT designates either a double bond or a single bond. y

A subject of the present invention is also the : use of a flavonoid and in particular of a compound of ) 30 formula I as defined above, for manufacturing a medicine intended to interfere (by = induction or inhibition) with the generation of clonogenic cells in tumours during treatment with at least one cytotoxic agent.

Ce od 2 - 5 - :

In the chemotherapeutic treatment of cancers with cytotoxic agents, flavonoids, and in particular _ the compounds of formula I, can be administered at the start of the chemotherapeutic treatments, either in a single dosagé intake or over several days at the start of these treatments (for example for 5 to 7 days) and, depending on the .chemotherapeutic protocol, at the start of each cycle of treatment (for example for 2 to 5 days) in the course of each therapy. . 10 . The compounds of formula I are advantageously administered by infusion (generally in 1 to 3 hours) at : doses of 5 to 50 mg/kg/day or 200 to 2000 mg/m?/day. - In order to obtain a maximum" effect on the production of clonogenic cells, the flavonoids should be administered such that the tissue concentrations obtained are as high as conceivably possible. " For protocols of treatment in the acute phases of therapies, the intravenous route is to be favoured using: ' ~- ready-to-use infusion solutes (bags, bottles, etc.) intended to be administered without modification + by intravenous infusion using an infusion line and according to the recommended flow rate: © = lyophilizates to be resuspended for intravenous infusion, using pharmaceutical solutes known to those skilled in the art; - for maintenance treatments, it is also —— possible to envisage the oral route when the . chemotherapy treatment favours the oral administration © 30 of cytostatic agents. For this purpose, oral lyophilizates (for oral or perlingual absorption) : immediate- or delayed-release tablets, oral solutions, . : suspensions, granules, capsules, etc. may be used. ’ The compounds of formula (I) are, for the most part, ‘compounds of natural origin, or are derivatives of compounds of natural origin. As examples, mention may be made of: 1) flavones such as: - quercetin,

Tae - 4-hydroxyflavone, - 6-hydroxyflavone, - 7-hydroxyflavone, - S5-methoxyflavone, © - 6-methoxyflavone, = 7-methoxyflavone,

Cs - 2-cyclohexyl-5-hydroxychromone,

Co - 2-cyclohexyl-6-hydroxychromone, - 2-cyclohexyl-7-hydroxychromone, : - wogonin or 5,7-dihydroxy-8- : methoxyflavone, - acacetin or 5,7-dihydroxy-4‘- : methoxyflavone, : - : ~ pedalitin or 5,6,3’,4’~tetrahydroxy-7- 5 methoxyflavone, : co ~ apigenin or 5,7,4’-trihydroxyflavone, - luteolin or 5,7,3",4'- ol _tetrahydroxyflavone, —- baicalein or 5,6,7-trihydroxyflavone, - - scutellarein or 5,6,7,4'- : tetrahydroxyflavone, oo - fisetin or 7,3’,4'-trihydroxyflavonol, — .robinetin or 7,3’,4’,5"~ tetrahydroxyflavonol, 25° - kaempferol or 5,7,4'- trihydroxyflavonol, ~ kaempferide or 5,7-dihydroxy-4’- methoxyflavonol, : — ‘ . - morin or 5,7,2",47- - 30 tetrahydroxyflavonol, Co - myricetin or 5,7,3’,4',5"- ~ pentahydroxyflavonol, . 2) flavanolols such as: : - aromadendrin or 5,7,4’- trihydroxyflavanolol, : - fustin or 7,3’,4'- trihydroxyflavanolol, - hydroxyrobinetin or 7,3",4’,5'- tetrahydroxyflavanolol,

E i . ud .

I -7- . ~ taxifolin or 5,7,37,47- trihydroxyflavanolol,

Lo raigeminoersean [ss 08 trihydroxyflavanone, ©. Lo pri i I eve gh Sra gi ; © - hesperetin or 5,7,3/= © trinydroxyflavanons. _. Flavones are the preferred compounds. ~The Cytotoxic agents can be chosen from: 1) intercalating agents, in particular ! daunorubicin, epirubicin, idarubicin, i ! 15 zorubicin, aclarubicin, pPirarubicin, acridine, : mitoxantrone, actinomycin D, eptilinium acetate; : ii) alkylating agents chosen from platinum derivatives (cisplatin, carboplatin, oxaliplatin); iii) a compound chosen from the other groups of alkylating agents: ~- cyclophosphamide, ifosfamide, chlormetrine, : melphalan, chlorambucil, estramustine, - busulphan, mitomycin C, . —- nitrosoureas: BCNU (carmustine), CCNU (lomustine), fotemustine, Streptozotocin, ~ triazines or derivatives: Precarbazine, To : decarbazine, ) : "30 ~ pipobroman, ~~ ethyleneimines: altretamine, triethylenethiophosphoramide, nN iv) a compound chosen from the other groups of ’ antimetabolic agents: . = antifolates: methotrexate, raltitrexed, Co : - antipyrimidines: S-fluorouracil (5-FU), cytarabine (Ara-C), - hydroxyurea, }

>a od [oe -8-

B - antipurines: purinethol, thioguanine, : pentostatin, cladribine = inducers of cytotoxic nucleoside synthesis: ro oo +7 ‘geméitabine, :

Loon Sav) .& compound chosen from ‘the .other groups of [40 BL TL Yinchtalkalbids which disorganize the mitotic

LC Co spindle: vincristine, vinblastine, vindesine,

CL navelbine ~~ .

SY C10 ok ~ agents which block thé depolymerization of the mitotic spindle: paclitaxel, docetaxel - ‘agents which induce breaks in the DNA by : topoisomerase II inhibition: etoposide, : teniposide - ) ~ topoisomerase I inhibitors which induce cleavages of the DNA: topotecan, irinotecan, vi) a splitting agent, which fragments the DNA, : such as bleomycin, vii) one of the following compounds; plicamycin, L asparaginase, mitoguazone, decarbazine, viii) an anticancer progestative steroid: medroxyprogesterone, megestrol, ix) an anticancer oestrogenic steroid: diethylstilbestrol, tetrasodium fosfestrol, x) an anti-oestrogen: tamoxifen, droloxifen, raloxifen, aminogluthetimide, xi) a steroidal anti-androgen (ex cyproterone) or a nonsteroidal anti-androgen (flutamide, TO nilutamide). - . ’ © 30 In particular, the compounds of formula I can be combined with all treatments with the major

Cytotoxic agents used in polychemotherapies for solid tumours, such as: BEFTSR : - alkylating agents: oxazophorines oo (cyclophosphamide, ifosfamide, chlorambucil, SPI melphalan) Co CE - nitrosoureas ER : - mitomycin C

EA TTT Ent EEA RA RY Sn

. : - antimetabolites, such as methotrexate, 5-FU,

Ara-C, capacitabine ~ agents which interfere with tubulin: vinca- alkaloids (vincristine, vinblastine, vindesine,

S navelbine), taxoids (paclitaxel, docetaxel), epipodophyllotoxin derivatives (etoposide, ‘teniposide) : : - bleomycin : : - topoisomerase I inhibitors: topotecan, irinotecan.

Similarly, the compounds of formula I can be combined with treatments with the major cytotoxic agents used ‘in oncohaematology for the treatment of blood cancers: Co N : 15 - Hodgkin’s disease: cyclophosphamide, mechlorethamine, chlorambucil, melphalan, ifosfamide, etoposide, doxorubicin, daunorubicin; ~ acute leukaemias: methotrexate, 6- mercaptopurine, cytarabine, vinblastine, vincristine, doxorubicin, daunorubicin, I~ asparaginase; i ~ non-Hodgkin’s malignant lymphomas: mechlorethamine, chlorambucil, cyclophosphamide, melphalan, ifosfamide, methotrexate, cytarabine, vinblastine, vincristine, etoposide, doxorubicin, daunorubicin, carmustine, lomustine, cisplatin; TT = chronic lymphoid leukaemias: mechlorethamine, chlorambucil, cyclophosphamide, melphalan, ifosfamide. Co

Results of pharmacological trials demonstrating : the properties of the compounds of formula (I) used j alone or in combination with cytotoxic agents will be : 35 given hereinafter. 1- "Interaction (stimulation or inhibition : of proliferation) with the generation of clonogenic cells (clonogenic assay)

wr

The assay 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 considered to be clonogenic if it has the capacity to proliferate and to give rise to a -cell colony. Human tumour stem cells are the cells behind the neoplastic cells which constitute a given tumour. These tumour stem cells are responsible for the processes of recurrence which can be observed after surgical } : 10 resection of primary tumours, and are also responsible for the formation of metastases. In a tumour. or a : tumour cell line, these clonogenic stem cells differ : from the other cells of the tumour or of the neoplastic cell line under consideration, by the fact that they conserve their capacity to proliferate in the absence : of any solid support. : In this assay, the tumour cells are cultured on a semi-solid support consisting of agar. Only cells which do not require a solid support for their growth (i.e. the very tumorigenic cells termed “anchorage- independent cells” by M.I. Dawson et al., Cancer Res. : ~ 1995; 55: 4446-4451; also named clonogenic cells with . reference to “clonal growth”) are capable of developing on such an agar-based support. Specifically, on such a medium, normal cells - which grow in “adherent mode” (“anchorage-dependence = cells” according to the terminology of M.I. Dawson) =~ such as, for example, - fibroblasts, do. not survive. Within a . tumour cell —_— : population, cultured on such a support, it is these ) 30 clonogenic cells (associated with an unlimited number of cell divisions, and the proliferation of which is termed “anchorage-independent fclonal] growth” by } M.I. Dawson) which are capable of growing. The } percentage of these clonogenic cells within a tumour or a cell line various between 0.1% and 0.001%. The nonclonogenic cells (associated with a limited number : of cell divisions) do not develop in this assay since they require a solid support for their growth which must take place in “adherent mode” (“anchorage- .

ww Prd a! - 11 ~ ’ dependent [adherent] growth”, according to M.I. 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 culturing, for example, the mammary tumour lines MCF7 and MXT, and the colorectal line HT-29, on the semi-liquid culture ‘medium termed “soft agar” was measured. On such a a ‘medium, only the clonogenic cells termed “anchorage- } : . independent (clonal) cells” by M.I. Dawson survive and develop. The growth of these cells in such a “nonadherent” mode indicates their degree of

A. tumorigenicity. The inhibition of the growth of the size of a tumour in which a greater number of » ‘clonogenic cells have developed then becomes the sign of reinforced cytotoxic activity. ) oo

Conversely, this assay can also reveal that a compound is. capable of inhibiting the generation/ proliferation of «clonogenic cells, which makes the tumour less able to develop, and therefore decreases : the population of tumour cells.

The tumour cell lines studied are maintained in . culture. in 25 cm? falcon dishes. They are then . trypsinized and the cells are well dissociated from : each other. The percentage of live cells is determined after trypan blue staining. A cell suspension at the concentration of 5 x 10% to 15 x 10% cells/ml (depending : on the cell type under consideration) is prepared in a solution of agar at 0.3%. Next, 200 ul of this —— , | suspension are seeded into petri .dishes 35 mm in ) 30 diameter, into which 3 ml of a base layer consisting of : a solution of agar at 0.5% are placed. The 200 pl of cell suspension are themselves covered with 1.8 mL of : ] . "an upper layer consisting of a solution of agar at } 0.3%. The dishes are then placed in an incubator at 37°C, 5% Co, and 70% humidity until treatment. The latter is carried out approximately 1 to 2 hours after seeding. The compounds to be assayed are prepared at a concentration 100 times greater than the concentration desired, and 50 pl of these treating solutions are tet - 12 - : deposited onto the upper agar layer of the

R Corresponding dishes. In the bresent study, the final concentration of the products assayed is 107%, 107 and 10° M. The dishes are then maintained for 21 days in an incubator. On the 21st day, the dishes are treated by depositing on the upper layer 100 pl of a solution of

MTT (3-(2,5-dimethylthiazol-2-yl)-2,5-diphenyl- ~ tetrazolium bromide) at 1 mg/ml Prepared with RPMI 1640 medium, for 3 h at 37°C. After this lapse of time, the cell colonies are fixed by adding 2 ml of formol per dish. After fixing for 24 hours, the formol is evaporated off and the number of cell colonies stained, and therefore consisting of cells which are metabolically active and which have. a surface area greater than 100 pm’ is determined using an inverted microscope.

The mean number of clonogenic cel} clones, determined for each experimental condition studied, is expressed as a percentage with respect to the mean number of clonogenic cell clones counted in the control condition and taken to be equal to 100%. These values, expressed as a percentage with respect to the control © condition, are given in Table 1, for quercetin.

TABLE 1

CELL LINES

I T= A A - * * NS i * % . . - * % NS - The results given in this table represent the mean values + the standard error of the mean (SEM) established on at least § cupules - Control condition = 100%

; ] yy @ i : hy - 13 -

IN = (NS: p>0.05; *: p<0.05; ** p<0.01; *=*: p<0.001).

On the three cell lines MCF7, HT-29 and MXT, ~ Quercetin is capable of partially inhibiting the

Lo 5. proliferation of the = clonogenic. cells. within ‘the _- obtained in the control condition (from 20% to 50%), - and therefore contributes to making the tumours from oo . ° 10 which they - are derived more sensitive to ‘the conventional treatment with cytotoxic agents. } 2 - Cytotoxic activity on .the nonclonogenic cells: “MTT assay” - )

The influence of the compounds of formula (I) on the nonclonogenic cells was evaluated using the MTT colorimetric assay. : The principle of the MTT assay is based on the mitochondrial reduction, by metabolically active live cells, of the vyellow-coloured product MTT (3-(4,5- dimethylthiazol-2-yl)-2,5~diphenyltetrazolium bromide) into a blue-coloured product, formazan. The amount of : formazan thus obtained is directly proportional to the amount of live cells present in the culture well (s).

This formazan amount is measured by Spectrophotometry.

The cell lines are maintained in monolayer

Co culture at 37°C in closed-cap dishes containing MEM 25 ‘ MM HEPES basic medium (Minimum Essential Medium). This TT medium is suitable for the growth of -a range of varied ’ : "30 diploid or primary mammalian cells. This medium is then supplemented with: : - a 5% amount of FCS (foetal calf ' serum) - decomplemented at 56°C for 1 hour, : - with 0.6 mg/ml of L-glutamine, - with 200 IU/ml of penicillin, : = with 200 pg/ml of streptomycin, = with 0.1 mg/ml of gentamicin.

) 2 > 4 ’

LL ; - 14 - . The 12 human cancer cell lines which were used : were obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA).

TE ‘These’ 12 céll lines are: -

Se So = U-373MG (ATCC ode: HTB=17 );'and U-87TMG (ATCC i ~~ BS49' (ATCC code: CCL-185) and A-427 (ATCC “010 0% 7 codé: HTB-53) which are two non-small-cell lung cancers, SOE ’ © - HCT-15 (ATCC code: CCL-225) and LoVo (ATCC SE code: CCL-229)y which are .two colorectal cancers, - ) . - T-47D (ATCC code: HTB-133) and MCF7 (ATCC : code: HTB-22) which are two breast cancers, - J82 (ATCC code: HTB-1l) and T24 (ATCC code:

HTB-4) which are two bladder cancers, - PC-3 (ATCC code: CRL-1435) which is a prostate cancer.

In .experimental terms: 100 pl of a cell : suspension containing 20 000 to 50 000 (depending on the cell type used) cells/ml of culture medium are seeded into flat-bottomed 96-well multi-well plates and . are incubated at 37°C under an atmosphere comprising 5%

CO; and 70% of humidity. After 24 hours of incubation, the culture medium is replaced with 100 pl of fresh — medium containing either the various: compounds to be ; © 30 tested at concentrations ranging from 1075 to 10-10 M, or the solvent which was used in order to dissolve the products to be tested (control condition). After... - . ooo 72 hours of incubation under the above conditions, the 7 - ) : culture medium is replaced with 100 pul of a yellowish solution of MIT dissolved, at a rate of 1 mg/ml, dn whos

RPMI 1640. The microplates are re-incubated for 3 hours at 37°C, and then centrifuged for 10 minutes at 200 g. Co

The yellowish solution of MTT is removed and the blue Co formazane crystals formed at the cellular level are

) : B a :

Lr ; - 15 = f dissolved in 100 pl of DMSO. The microplates are then : shaken for 5 minutes. The intensity of the blue coloration = resulting, therefore, from the

EE ‘transformation BE ‘the yellow MTT product into blue

SE = formazane by .the cells still. alive at-~the end of the = ‘wavelengths of 570 nm and 630 nm corresponding, © respectively, to the wavelengths of maximum absorbence ©." 10" “of the ‘formazane and to the background noise. Software "integrated into the spectrophotometer calculates the mean’ values of optical ‘density, and the standard - deviation (std. Dev.) and standard error of the mean - (SEM) values. - :

By way of example, the results of the mean Co optical density, expressed as percentage with respect to the mean optical density measured in the control condition (taken to be equal to 100%), obtained - by way of nonlimiting example - with a flavonoid: quercetin, on the 5 tumour cell lines U-373MG, T24,

LoVo, MCF7 and A549, will be given in Table II.

TABLE II cows 3.1 NS 2.2 NS 2.9 NS 2.4 NS 3.2 NS 1.7 NS 0.7 NS 1.7 NS 2.0 NS 3.8 NS 2.0 NS 2.4 *+* A . 3.1 NS 2.2 NS 4.7 NS 2.1 NS 3.0 NS 4.2 * 2.5 *%+ | 2.3 wu* 2.1 NS 3.8 Ns 2.0 ** 2.988 | - EE - ’ 3.6 * 3.7 NS 2.3 NS 3.5 Ns 2.2 NS 1.7 NS - Concentrations expressed in mol.l7 : oo } } J A Sn 3 - xx * yy = mean value standard error of the mean N CTS - Control conditions = 100% B - (NS/p > 0.05; *: p < 0.05; **: p < 0.01; p < 0.001). -

so» 3

ER - 16 - . These results show that quercetin has weak : antitumour power. This product, which is non-cytotoxic, induces inhibition of the overall cell proliferation of ‘these “lines only at the concentration of 107° M, and . .....5. this vinhibition does. not exceed 20%. At the other ~ .o. . . . 73.7 ="Determination of the maximum tolerated

CUTE 10 dose (MTD): EEA

The evaluation of the maximum tolerated dose was carried out in .4- to 6-week-old B6D2F1/Jico mice. : The compounds were administered via the intraperitoneal : route at increasing doses ranging. from -2.5 to 160 mg/kg. The value of the MTD (expressed in mg/kg) is determined based on the observation of the survival rate of the animals over a period of 14 days after a

To single administration of the product under consideration. The evolution of the weight of the animals is also monitored during this period. When the value of the MTD is greater than 160 mg/kg, the value of the MTD is assimilated to 160 mg/kg by default.

The quercetin is assocated by default with an

MTD .equal to 160 mg/kg. This result emphasizes that the products belonging to the flavonoid family have no direct toxicity and can be used at high tissue concentrations, and therefore at high doses.

Examples of methods of use of the compounds of —— formula (I) in protocols of mono- or polychemotherapy ) 30 with cytotoxic agents will be given hereinafter.

A. Solid tumours i 1°/ Lung cancers 1.1 non-small-cell lung cancers (advanced stage): Cg i 1 R 3 ro - 17 - . - added to the recommended protocol (T. Le Chevalier et al., J. Clin. Oncol. 1994; 12: 360-367) are the intravenous infusions of a compound of formula I:

Cl _ RA Leal SRE Co a a Dye and Da IE

SA

This therapy is to be repeated 8 times. 1.2 small cell lung cancers (advanced stage): - added to the recommended CAV or VAC protocol (B.

J. Roth et al., J. Clin. Oncol. 1992; 10: 282-291) are the flavonoid infusions: [| dose | route | days

EEE or 5-50 mg/kg/day i.v. 1 h infusion bolus : bolus : i e vincristine 1 to 1.4 mg/m? i.v. D,

ETT

(max 2 mg)" - 15 This therapy is to be repeated 6 times every 21 ’ days. - added to the recommended Pt-E protocol (B.J. Roth oo et al., J. Clin. Oncol. 1992; 10: 282-291) are the flavonoid infusions:

BE y nn # i :

JR - 18 - * flavonoid 200-2000 mg/m*/day oo D, - Dy 1 ‘or 5-50 mg/kg/day | i.v.

ES UA SOU SE Ft 1 hinfusion

EE EEE tl I SRE J BY SP 60 mings : oe | | irifusion

EA | 6b minute infusion | i each cycle is repeated every 21 days, and the . therapy comprises 6 cycles. - - - 1.3. locally advanced or metastatic non-small-cell bronchial cancer: i e¢ monochemotherapy dose | route | days

I EE or 5-50 mg/kg/day i.v. then 1 week/rest 1 h infusion : : 0.5 hour infusion then 1 week/rest the therapy possibly comprising the repetition of this 4-week cycle. : oT e gemcitabine/cisplatin combination:

’ i 3 .

Cer - 19 - dose | route | gays

I | or 5-50 mg/kg/day | i.v.

Re Rake _ BS cu} 0.5 Hout infusion 2 SEPA SREP Sa SE ] lg : SOMERS a | « cisplatin BEEN Toh S/n aay — TE FT or Ea ? Le oo 20-60 minute iv.

CL infusion — . . the therapy comprising the repetition of this oo cycle every 21 days. Loe 2°/ Breast cancers - To - CMF protocol as -an adjuvant treatment of operable breast cancer (G. Bonnadonna et al., N. Engl. J.

Med.; 1976; 294: 405-410): dose route| days "|e flavonoid 200-2000 mg/m*/day Dy to Dig ei : 1 h infusion bolus . each cycle is repeated every 28 days, and the : therapy comprises 6 cycles. a : 15 - AC ‘protocol (B. Fisher et al., J. Clin. Oncol.; i } 1990; 8: 1483-1496) as an adjuvant treatment: CL

EE - 20 - [dose [route] gays

I a er or 5-50 mg/kg/day | i.v. Dy "1 h infusion

Lo _ i bolus each cycle is repeated every 21 days, and the - therapy comprises 4 cycles. wl - breast cancers with metastases: . - in the FAC protocol (A.U. Buzdar et al., Cancer . 1981; 47: 2537-2542) and its various adaptations, : the flavonoid infusions are added according to the following scheme (nonlimiting): dose | route | days se flavonoid 200-2000 mg/m*/day D, - Ds and ial Er : 1 h infusion or Dy = Dg : bolus Dy - D : .. bolus D; and D: e cyclophosphamide © 500 mg/m? oral or D; ome em . bolus : each cycle is repeated every 3 weeks until a RE . ‘new progression of the disease is diagnosed. Co : 15 - in the CAF protocol (G. Falkson et al., Cancer ED 1985; 56: 219-224): ERR we Te? : oo y - 21 - * flavonoid 1200-2000 mg/m’/day Dy - Dy

SE or 5-50 mg/kg/day | i.v.

EE CRT 1 hinfusion g each cycle is repeated every. 28 days until a new progression of the disease is diagnosed. - in the CMF protocol: Co = ose | route | gays

EEE

} or 5-50 mg/kg/day i.v. Dg - Diz 1 h infusion bolus bolus bolus : this cycle is to be repeated every 3 to 5 — weeks, and the therapy comprises 6 cycles. . 10 . ’ - in the CMF-VP protocol: ;

JRE N ’ ik » - 22 - * flavonoid 200-2000 mg/m?/day D; - Ds

RE | or 5-50 mg/kg/day | i.v. Ds - Diz : 0 | 1h infusion | Dis - Dig

ES SI SA SDE SPIRE NA es Das.’

RE LC _ mg/m’/day SL D2; 1.2 mg/m?/day Das

Dio this therapy is to be repeated every 4 weeks. ) = in the FEC protocol: . dose | route | days [REE : or 5-50 mg/kg/day i.v. Dg - Di; ’ } 1 h infusion this therapy is to be repeated every 3 weeks. ’ - in the MMC-VBC protocol (C. Brambilla et al., © 10 Tumori, 1989; 75: 141-144):

.

JT : - 23 - : dose | route | days e flavonoid 200-2000 mg/kg/day D, - Ds and or 5-50 mg/kg/day | i.v. | Dy - Dis

Ea 1h infusion ‘bolus ‘this therapy is to be repeated every 28 days until progression of the disease is diagnosed. Er - in the NFL protocol (S.E. Jones et al., J. Clin. © Oncol. 1991; 9: 1736-1739): Co ) dose [route | days e flavonoid © 200-2000 mg/m?/day D; - Ds

EET

1 h infusion bolus e 5-FU 1000 mg/m’ i.v. D; - Ds; 2 : infusion bolus the therapy comprises two cycles 21 days apart, oT - 10 and then requires an evaluation. :

The flavonoid infusions can also be combined with the treatment of breast cancers with metastases, oo - when a taxoid is used, for example: ) 15 - with paclitaxel (F.A. Holmes et al., J. Natl.

Cancer Inst. 1991; 83: 1797-1805) in the treatment of forms with metastases possibly resistant to anthracyclines:

aR ; - 24 - dose | route | days e flavonoid 200-2Q00 mg/m’/day Dy - Ds “or 5-50 mg/kg/day i.v. a Lb "1 n infusion

SR eo paclitaxel : oo 175 tng /mi® iv. | 'p, i he - oo SET Ci Infusien” ib Lo RR

CL This cycle is repeated every 21 days until a 7 new progression of the disease is diagnosed. - with docetaxel (C.A. Hudis et al., J. Clin. Oncol. 1996; 14: 58-65), in locally advanced or metastatic breast cancer which is resistant or in relapse after cytotoxic chemotherapy (having : - comprised an anthracycline), or in relapse during an adjuvant treatment: [dose | route | gays

EEE RE or 5-50 mg/kg/day i.v. : 1 h infusion © | * docetaxel 100 mg/m’ i.v. D; or 60-100 mg/m’ in a 1 hour (or 24 hour) infusion

This cycle is repeated every 21 days for a 2- . cycle therapy, or until the appearance of progression - 15 of the disease. : - in dose increase protocols, combining transplant . of autologous medullary cells and of peripheral ’ blood stem cells, as reinforcement for the first choice treatment, for example:

In . : - 25 - : = CPB protocol (W.P. Peters et al., J. Clin. Oncol. : 1993; 11: 132-1143) in which the i.v. infusion of stem cells takes place on days D.i;, Dg and Dj:

I SE _ 1h infusion + cyclophosphamide |. 1875 mg/m to | iv. | Dy to D. : : : infusion i * cisplatin | 55 mg/m’/day i.v. D.s to D4

I a Wd : continuous infusion e carmustine : 600 mg/m’/day iv. D_; oo (BCNU) in a 2 hour h infusion : 5 = CICb protocol (K. Antman et al., J. Clin. Oncol. 1992; 10: 102-110), in which the i.v. infusion of stem cells takes place on Dg:

r - 26 - dose | roure | aays eo flavonoid 200-2000 mg/m?/day D.; to D.

N or 5-50 mg/kg/day i.v.

IRE oC _ _ : 1h infusion _

So. |*erclovhosphamide | 1500 mg/m | i.v. | by co bg fn BEATE Ri Te continuous infusion ls | (4 doses) _ wel Coe tniotepa | 12s mgm? | iv. | boron

Se Ce in a 24 hour . i continuous infusion : (4 doses) e carboplatin 200 mg/m? CiLv. .D.; to D-; i | in a 24 hour : continuous infusion (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, | 1924 (Suppl 18B): 95) in which the i.v. infusion of haematopoietic stem cells oo takes place on Dg: dose | roure | ams e flavonoid 200-2000 mg/m*/day 3B Ds to D_; or 5-50 mg/kg/day i.v. 1 h infusion J * cyclophosphamide 1500 mg/m*/day D.s to D_; . . in a 1 hour infusion : e thiotepa 150 mg/m*/day D.s to Ds e mitoxantrone 10-15 mg/m’ iv. D.s LO D.s - : in a 1 hour infusion 3°/ Gynaecological cancers 3.1 Ovarian cancer:

RE - 27 - - for the treatment of ovarian carcinomas, in

Particular metastatic ovarian carcinomas: : i) BAC protocol (G. A. Omura et al., J. Clin.

Cee 008 Oncoli 1989; 7: 457-465): the flavonoid infusions

RE Ul. ere administered . according to the following

Capi FRR : schem po SEE BN EE oe e flavonoid 200-2000 mg/n’/day. | | bp, - py or 5-50 mg/kg/day | i.v. : 1 h infusion ® cisplatin 50 mg/m’ [7] D, (or 40-90 mg/m?)

Len TT ® doxorubicin 50 mg/m? bolus D; a SAE ER NE * cyclophosphamide 1000 mg/m’ i.v. D; 1 to 2 hour infusion BN (or 200 to 600 mg/m°) 10. this cycle is repeated every 21 to 28 days, and the therapy comprises 8 cycles. ii) altretamine protocol, according to A. Marietta et al., (Gynecol. Oncol. 1990; 36: 93-96): ee } e flavonoid 200-2000 mg/m?/day D; - Ds : or 5-50 mg/kg/day iv. Dg - Di; 1h infusion ¢ altretamine | 200 mg/m?/day oral D: ~ Di: - divided into 4 doses the therapy comprising two cycles, 28 days : : apart.

Rn : = 28 = . : ii) paclitaxel protocol: the flavonoids can be added to the paclitaxel protocol as has been described by W.P. McGuire et al., (Ann. Intern.

Med. 1989; 111: 273-279): : * flaversia | 200-2000 mg/m’/day : or 5-50 mg/kg/day i.v. oo 1 h infusion ® paclitaxel | oo 135 mg/m? hE 3 hour or 24 hour "the therapy comprising two of these cycles, 28 days apart (with evaluation at the end). - for the treatment of metastatic and resistant ovarian carcinomas, the flavonoids can be added to the second choice protocol, based on topotecan: or 5-50 mg/kg/day : . 1 h infusion 0.5 hour infusion the cure comprising two cycles, 21 days apart (with evaluation at the end) : To , according to A. P. Kudelka et al. (J. Clin. Oncol. ) 1996; 14: 1552-1557). oo 3.2 Trophoblastic tumours: i : - in patients at low risk, the flavonoids may be : combined with the protocol described by : H. Takamizawa et al. (Semin. Surg. Oncol. 1987; 36-44):

bE : * flavonoid 200-2000 mg/m?/day i.v. | D,-Ds pe or 5-50 mg/kg/day = 1 h infusion 2 . methotrexate MTX) | 20 mg/day | im. | p,-Dy

E * dactinomycin | 0.5 mg/day in a bolus | i.v. D;=Ds

E Co ol iemeny a : (MTX-DATC protocol). ; © 3.3 Cancers of the uterus: : = the flavonoids can also be combined with the Cav : : (or VAC) Protocol according to the- Scheme below: a | __ dose Tone ane 200-2000 mg/m?/day . ° 1 . flavonoid or 5-50 mg/kg/day 1 h infusion i.v. D1=Ds . 750-1200 mg/m? , - 2 ln an infusion vneristioe | uae iw [oT the therapy comprising the repetition of this cycle every 21 days. : « = in the FAP protocol: * flavonoid 200-2000 mg/m/day : or 5-350 mg/kg/day i.v. D;-Ds : 1 h infusion * fluorouracil] 600 mg/m?/day i.v. Di, Dg (5-FU) — * doxorubicin 340 mg/m’ iv. | bp,

ET EE T= oe BP : the therapy comprising the repetition of this cycle every 21 or 28 days.

]- 4°/ Cancers of the testicle and of the prostate 1 = the flavonoids can also be combined with

E Protocols for cancer of the testicles: ] oT ~ 'BEP protocol: : | dose [oT a] ] * flavonoid 200-2000 mg/m?/day or 3-30 mg/kg/day | i.v. | p,-p, 1 h infusion ] 2 20 mg/m® iv. Dy in an infusion ! p re 2 7. in an infusion - © cisplatin | 0 moira the therapy comprising 3 cycles, at a rate of 1

Cycle every 21 days. : 5°/ Cancers to the bladder ——== =0 the bladder =~ the flavonoids can be combined with the CIsca2 (also called PAC) protocol: ~ 15

SR fi hi * flavonoid 200-2000 mg/m?/day or 5-50 mg/kg/day iv. D;-D; : 1 h infusion eteplatin TT shmem Tip * cyclophosphamide 600 mg/m? i.v. . in an infusion } * doxorubicin 75 mg/m? i.v. D; in an infusion : the cycle being repeated every 3 weeks. = in the MVAC protocol (according to CN Sternberg et

I., J. Urol. 1988; 139: 461-469) :

EE - 31 - * flavonoid = | 200-2000 mg /m?/day D;~Ds : or 5-50 mg/kg/day i.v. Dis—Dis 1 h infusion _ D33=Das ’ 5 Lo : _ B _ I D,, Dis, Daz * cisplatin 70-100 mg/m? iv. |'D, or D; 1 h infusion . - : this cycle being repeated every 4 to § weeks, for 2 minimum of 2 cycles. - . 5 - - ) 6°/ Nasopharyngeal carcinomas/head and neck cancers - The flavonoids can be validly combined with the

Co polychemotherapy protocols used in the treatment of these cancers: 6.1 Nasopharyngeal cancers: : ~- ABVD protocol: e flavénoid 200-2000 mg/m’/day Dy~Ds or 5-50 mg/kg/day iv. Dg—Dio ‘1 h infusion or Dj;s=Di ——— e doxorubicin 30 mg/m*/day Cl ilv. D; and Dg , ’ or Dis

Co e bleomycin 10 mg/m?/day i.v. D; and . - Dg or Dis e vinblastine 6 mg/m?/day i.v. D, and : : Dg or Dis ] oe dacarbazine 200 mg/m?/day i.v. | Dp; and ; : Dg or Dis : the therapy comprising 1 to 6 cycles repeated at a rate of 1 cycle every 4 weeks.

EEC. - 32 -

In 6.2 Head and neck cancers with metastases:

E = in the Pt-FU protocol (ex: for cancers of the : pharynx): according to the DVAL Study Group (New 5 Engl. J. M. 1991; 324: 1685-1690): 3 * flavonoid 200-2000 mg/m*/day

El : or 5-50 mg/kg/day i.v. D1~Ds 3 1 h infusion 3 100 mg/m? | iv. D;

E 1 hour infusion ; ¢ fluorouracil (5-FU) 1000 mg/m*/day i.v. D;~Ds ; continuous infusion : the therapy comprising two cycles at a rate of 1 cycle every 3 weeks. oo 7°/ Soft tissue sarcomas ——— —=S5Ue sarcomas —- The flavonoids can be introduced into a protocol such as the CYVADIC protocol: ~ according to H.M. Pinedo et al. (Cancer 1984; 53: 1825); dose Trowe] wp] e flavonoid 200-2000 mg/m?/day D;~Ds : or 5-50 mg/kg/day | i.v. Dg-Dig a lh infusion D15-Di4 * cyclophosphamide (Cy) 500 mg/m? bolus * doxorubicin (a) 50 mg/m’ bolus ive | me * dacarbazine (DIC) 250 mg/m*/day i.v. D;~Ds : [15 minute infusion : the therapy comprising the repetition of this cycle every 4 weeks, for 2 cycles at first.

4 - 33 - iE 8°/ Hormono-resistant prostate cancer, with metastases z = in the VBL-estramustine Protocol, according to ; G.R.

Hudis et al. (J.

Clin.

Oncol, 1992; .10: 1754: 3 5 1761) 4 * flavonoid 200-2000 mg/m?/day D;~D3, Dg=Dyq : or 5-50 mg/kg/day | i.v.

Dis-Dy5, D3y~Dyy : 1 h infusion D29=D3;,. D3s-Dsg * estramustine 200 mg/m? tid oral | each day for 6 (600 mg/m?/day) weeks : : a treatment cycle lasting 6 weeks and being followed by a free period of 2 weeks. 10° 9°/ Germ cell cancers : —— ==-2 cancers i) for tumours with a favorable prognosis: = Pt-E protocol, according to G.J.

Bos] et al. (J.

Clin.

Oncol. 1988; 6: 1231-1238): e flavonoid 200-2000 mg/m?/day or 5-50 mg/kg/day i.v.

D1-Ds 1 h infusion e cisplatin 20 mg/m’/day i.v.

D;~-Ds — to 60 minute infusion * etoposide (E) 100 mg/m®/day 1oilv.

D;-Ds ) 1 hour infusion : the therapy comprising 4 cycles, at a rate of 1 20 cycle every 21 or 23 days. ) ii) for tumours with metastases: : ~- PEB protocol, according to S.D.

Williams et al. (N.

Eng.

J.

Med. 1987; 316: 1435-1440) ;

N hel J = 34 = * cose [route] days e flavonoid 200-2000 mg/m?/day D;-Ds or 5-50 mg/kg/day | i.v. Dg-D11 1 h infusion D;s-Dig e cisplatin (P) 20 mg/m?/day i.v. D;-Ds 20 to 60 minute infusion - eo etoposide (E) 100 mg/m*/day i.v. {Ds,Dg,Di6 : 1 hour infusion ® bleomycin (B) 300 (or mg) /day i.v. D,~J5 . bolus the therapy comprising 4 cycles, at a rate of 1 cycle every 21 days. 10°/ cancers of the kidney oo - - metastatic renal carcinoma: the flavonoids can be introduced into the protocol described by M. J.

Wilkinson et al. (Cancer 1993; 71: 3601-3604): : 10 dose [route | days e flavonoid 200-2000 mg/m*/day D,-Ds or 5-50 mg/kg/day i.v. Dg=Dys } 1 h infusion ~|* floxuridine 0.075 mg/kg/day i.v. | Dy;-Diya continuous infusion the therapy comprising two cycles, 28 days apart. - nephroblastoma: the flavonoids can be introduced —_— into the DAVE protocol: dose [route] days eo flavonoid 200-2000 mg/m?/day D;~Ds; or 5-50 mg/kg/day | i.v. Dg-Dio . 1 h infusion e cyclophosphamide 200 mg/m?/day i.v. D,, Dg 1 hour infusion at a rate of one cycle every 3 to 4 weeks. :

TT - 35 -

HR! ro 11°/ cancers of the digestive tract , re 11.1 Cancers of the ©esophagus

PETRCred = ~ the flavonoids can be introduceq into the Fap

EE Protocol according to.

Sh ag. ih * flavonoiqg 200-2000 ng/m*/day D;-D;

I 1h infusion 3 : J S-fluorouraci] (5-Fu) 600 mg/m? . ) EAT

LE this cycle being repeated Svery 3 to 4 weeks. 11.2 Stomach Cancers ~ in gastric Carcinomas which are advanceqg and/or with Metastases: — EAPp Protocol} (according to p, Preusser et : al., g. Clin. oOncoj. 1989; 7. 1310): * flavonoig 200-2000 mg/m*/day —_— : or 5-5p mg/kg/day D1=Ds, Dg-p,, : 1h infusion * etoposide 120 mg/m*/day D3, by, D5 of : : 1 hour infusion D,-Dq * cisplatin 40 mg/m*/day 1 hour infusion - at a rate of 1 Cycle Every 28 days. - FAMtx Protocol]: according to J.a. Wils et al. (J. Clin. Oncol. 1991; gg. 827):

EEE - 36 -

Po to | * flavonoig 200-2000 mg/m*/ day

OL 3°50 mg/kg/day 2 1h infusion ee * fluorouraciy (5-FU) 1500 mg/m? bolus 15 (F) 1 hour after tye 15 methotrexate ia * doxorubicin (a) 30 mg/m? bolus EE 4 Fi pod . methotrexate (Mtx) 1500 mg/m?

EF 30 minute infusion 2 rE the therapy Comprising twe cycles at first, 2g a days apart CL : = in Certain diseases, this Protocol op its 1 Variant (epirubicin replacing doxorubicin) may be used according to the following Scheme . : * flavonoig 200-2000 mg/ ; m*/day ] or 5-50 mg/kg/day 1 h infusion * doxorubicin (A) 30 mg/m* bolus or epirubicin (A) 60 mg/m? bolus ® methotrexate . (to be infused before —_— the s5-py) ’ 12e/ Colorecta] Cancersg - the flavonoids can be introducey into the ] Protocol for FU-Levamisole adjuvant treatment of : 15 colorecta] Cancer (according to C.a. Moerte; et ~ al., nN. Eng. 7. Med. 1990; 323. 352).

i a. ’ ’ N dose Trowe] ame] e flavonoid 200-2000 mg/m?/day D;-D; : or 5-50 mg/kg/day i.v. ‘D29=-Ds; ] 1 h infusion * S5-fluorouracil | 450 mg/m?/day bolus EET * S-fluorouracil 450 mg/m? bolus (iv. | on] : * levamisole 50 mg tid oral 3 days/week : one week out : of two .

B the 5-FU bolus treatment being repeated each week after the D:1-Ds induction phase, for 52 weeks; the treatment with a flavonoid being repeated in the same rhythm, on the day of the 5-Fyu bolus and then on the following 2 days. = for the treatment of colorectal cancer, with metastases, which is resistant to treatment with 5-fluorouracil (5-FU) : - according to M.L. Rothenberg et a]. (J. Clin.

Oncol. 1996; 14: 1128-1135) :

I I e flavonoid 200-2000 mg/m*/day D;~Dj, Dg~Dy,, or 5-50 mg/kg/day i.v. Dy5~D;5, D25-D>, 1 h infusion ‘ ) the therapy comprising two cycles, 42 days apart. 13°/ Kaposi's sarcomas ] - the flavonoids can be combined with the two ’ protocols which use anthracyclines formulated inp : liposomes: 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):

SE oT - 38 -

Pol a IP ove Ry

N * flavonoid 200-2000 mg/m?/day D;-D; and or 5-50 mg/kg/day i.v. Di5-Dy5 ] 1 h infusion 4] * liposomal daunorubicin 20 mg/m*/day i.v. Di, Dis : j 1 hour infusion the therapy comprising two cycles I'epeated 28 days 1 apart, before evaluating the effects, 3 5 E

Hl ii) Protocol of M. Harrison et al, (J. Clin. 1

Oncol. 1995; 13. 914-920): = ] | cose ToT * flavonoid 200-2000 mg/m’/day D,-D, or 5-50 mg/kg/day i.v. l h infusion * liposomal doxorubicin 20 mg/m" i.v. D,

Co 30 minute infusion the cure comprising two cycles repeated 28 days apart, before evaluating the effects. 14°y Metastatic melanomas

B oo ——— - the flavonoids can also be incorporated into combined Protocols for treatment of metastatic malignant melanomas: -- - DTIC/TaMm Protocol: according to Gg. Cocconi et © 20 al. (N. Eng. J. Med. 1992; 327. 516), the therapy comprising the repetition of 4 Cycles, at ga rate of 1 cycle every 21 days, according to the Scheme below:

ol i - B

IN gi 3 i - - 39 = i . * flavonoid 200-2000 mg/m?/day 1 or 5-50 mg/kg/day D;-Ds 3 hy 1 h infusion : 7 * dacarbazine 250 mg/m*/day 3 2 (DTIC) infusion [15 to 30 min. if 3 2 central Catheter] D;-Ds

Ep or [30 min. if Periphera] = infusion in 250 ml] {id ®* tamoxifen 20 mg/m*/day

EE (TAM) 1 the therapy comprising 4 Cycles at 4 rate of 1 19 Cycle every 23 days. : g 5 : 15°/ Neuroendocrine carcinoma =~ flavonoids can be combined With the Protocol ! described by C.G. Moert] et al. (Cancer 1991; 6s: i 10 227): { = Pt-E Protocol: * flavonoid 200-2000 mg/m*/day or 5-50 mg/kg/day 1 h infusion * etoposide 130 mg/m*/day —— 1 hour infusion = 45 mg/m*/day } 1 hour infusion the therapy comprising two Cycles repeated &very 28 days.

E 16°/ cancer of the pancreas = advanced stage Pancreatic adenocarcinoma - the flavonoids can be combined with the gemcitabine treatment, according to the Protocol of M. Moore

& . Ie a ‘ le et al. (Proc. Am. Soc. Clin. Oncol. 1995; 14; 473) ; 1 |___ cose rows] ao

A } * flavonoid 200-2000 mg/m?/day D;-Ds, Dg-Dyp, Dis, 7 or 5-50 mg/kg/day i.v. Das, Dag, Ds, Dys, Ds. 3 1 h infusion : * gemcitabine 1000 mg/m? i.v. D1, Ds, Dys, Daz, Dag, / + 0.5 hour infusion D3s, Ds3, then Ds, ] E then once/week for 1 ‘ . 3 weeks, and then 1 week of rest and : evaluation : 5 B. Oncohaematolo To - : ——ofaematology 1°/ Adult acute leukeamias ——c---° -eukeamias 1.1 Acute lymphoblastic leukaemia: 1.1.1 Linker protocol

The flavonoids can be added to Linker Protocols —- induction chemotherapy and consolidation

Chemotherapy - . (see C.A. Linker et al. Blood 1987; 69: 1242-1248 ang C.A. Linker et al.

Blood 1991; 7s: 2814-2822) - according to the following schemes: i) induction chemotherapy: ese Trowel an e flavonoid 200-2000 mg/m*/day D;-Ds, or 5-50 mg/kg/day i.v. Dg~Dya, D15-Dy4 1 h infusion * daunorubicin 50 mg/m” bolus every 24 hours (30 mg/m? in | i.v. Di, Da, D, patients more than 50 - years old) :

EE NEE Ron re vey i ii) consolidation chemotherapy (treatment A):

i PY ry = v i. , : dose lowe] gage e flavonoid 200-2000 mg/m?/day D,-Ds, Dg~Di2 ) or 5-50 mg/kg/day iv. 1 h infusion ®¢ daunorubicin 50 mg/m? bolus i.v. D;, D2 ] every 24 hours * prednisone © 60 mg/m°/day oral D;-Diy divided into 3 doses *¢ L-asparaginase 12000 U/m? i.m. D2, D4, D7, Dg . and Dig consolidation therapy A comprises 4 consecutive cycles such as the one described above = .cycles 1, 3, 5 and 7. iii) consolidation chemotherapy (treatments B and C): ) The therapies described below correspond to consclidation cycles 2, 4, 6 and 8 (therapy B) and 9 (therapy C), described by C.A. Linker et al.:

Cgmeraby Bi | dose rows | ays e flavonoid 200-2000 mg/m‘/day D1~Ds, Dg=D; 4 oo or 5-50 mg/kg/day i.v. 1 h infusion e Ara-C 300 mg/m’ i.v. Dy, Dy, Dg, Dy; : 2 hour infusion : eo teniposide 165 mg/m” i.v. D1, Dy, Dg, Dy; — "2 hour infusion TT (4 cycles)

] i Pd [ad -

Eos

Ea - 42 - !

P therapy c: route | gays * flavonoid 200-2000 mg/m?/day ] D;-Ds ; Or 5-50 mg/kg/day iv. 1 h infusion i ®* methotrexate 690 mg/m? i.v. D;-D: j 42 hour continuous infusion * leucovorin : 15 mg/m* oral D,-Ds : every 6 hours 1.1.2 Hoelzer protocol ? The flavonoids may be added to the cytotoxic 5) 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: : * flavonoid 200-2000 mg/m’ /day Dy-Ds, Dg-D;5,

Or 5-50 mg/kg/day | i.v. Dis-Dyg 1 h infusion en il cy 1.5 mg /m* i.v. D1, Ds, D1s, Das (maximum 2 mg)

EE EES ES

* L-asparaginase 5000 U/m? i.m. oo (maximum 2 mq) TT : ii) induction chemotherapy/Phase 2:

Phase 2 of the induction may be carried out as follows:

HO | - 43 - * flavonoid 200-2000 mg/ D29-D33, D3g~Dyo, u ~ m?/day i.v. 'Dy3-Dys

H or 5-50 mg/kg/day 3 1 h infusion 3 * cyclophosphamide 650 mg/m? i.v. D29, Dy3, Dss : | {maximum 1000 mq) 3 ®* cytarabine 75 mg/m*/day i.v. D31-Ds;, D3g=Dy; ; 1 hour infusion Dys~Das, Ds=Dss : * methotrexate 10 mg/m*/day 1.v. | Diy, Ds, Dys, Das 4 (maximum 15 mg) bed i iii) reinduction chemotherapy/Phase 1: ; ese Troms] am * flavonoid 200-2000 mg/m*/day D1~Ds, Dg-D;.,, : or 5-50 mg/kg/day | i.v. D15=Dyg, Dy2=Dag 1 h infusion a a 1.5 mg/m°/day oral | Dj, Di, D5 and : (maximum 2 mg) Dz» iv) reinduction Chemotherapy/Phase 2:

I

* flavonoid 200-2000 mg/m*/day D31~D3s, D3g~Dys TT or 5-50 mg/kg/day i.v. 1 h infusion ; . * cyclophosphamide 650 mg/m" i.v. (maximum: 1000 mg) : ® thioguanine 60 mg/m’ oral D2g=Dy, 1.2 Acute myeloid leukaemias: 1.2.1. Treatment of adults of any age

§ ~- 44 - . The flavonoids can be added, according to the scheme below, to the treatment which incorporates the standard dose of cytarabine previously described by

R.0. Dilleman et al. (Blood, 1991; 78: 2520-25286),

Z.A. Arlin ‘et al. (Leukemia 1990; 4: 177-183) and

P.H. Wiernik et al. (Blood 1992; 79: 313-319): dose |route| days e flavonoid 200-2000 mg/m’/day D1~Di2 or 5-50 mg/kg/day i.v. 1 h infusion e cytarabine 100-200 mg/m?/day iv. D;-D; in a continuous infusion e daunorubicin 45 mg/m?/day as a bolus _| i.v. D,~D;,0r (30 mg/m*/day if age = 60) . Dg-Dio or } e mitoxantrone 12 mg/m" iv. D;-D; in a daily bolus or e¢ idarubicin 13 mg/m’ i.v. Dy-Dj in a daily bolus 1.2.2. Treatment of adults under the age of 60 : i) induction chemotherapy:

This induction cycle incorporates the administration of cytarabine at high dose according to the following scheme: e flavonoid 200-2000 mg/m*/day D;-Dyo - or 5-50 mg/kg/day i.v. - 1 h infusion e Ara-C (cytarabine) 2000 mg/m*/day i.v. D;-Dg in a 2 hour infusion, every 12 hours EE e Daunorubicin 60 mg/m’/day i.v. D,~Dg

N in a 24 hour : continuous infusion or e Cytarabine 3000 mg/m?/day i.v. D,-De in a 1 hour infusion, every 12 hours e Daunorubicin 45 mg/m* bolus i.v. Dy-Ds every 24 hours o (in order to reduce the risk of C.N.S. toxicity, in the event of renal failure, adjust the cytarabine dosage to the creatinin clearance) 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). ii) consolidation chemotherapy: . 10 The cycle, described hereinafter, will be repeated 8 times, at a rate of 1 cycle every 4 to 6 weeks (according to R.J. Mayer et al., N.

Engl J. Med. 1994; 331: 896-903). : dose |route]| aays e flavonoid 200-2000 mg/m" /day i.v. D,~Ds or 5-50 mg/kg/day 1 h infusion e cytarabine 3000 mg/m’ i.v. | Dy, Ds,Ds in a 3 hour infusion, every 12 hours (4 cycles) {then 100 mg/m°/day » cytarabine every 12 hours S.C. D;~Ds e daunorubicin 45 mg/m” bolus i.v. D, (4 cycles) iii) consolidation chemotherapy (with high dose of cytarabine):

The cycle, described hereinafter, must be TT . repeated twice and is adapted according to

G.L. Phillips et al. (Blood 1991; 77: 1429- 14335); 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):

dose Toute] cape eo flavonoid 200-2000 mg/m?/day i.v. D;-Dj or 5-50 mg/kg/day 1 h infusion ® cytarabine’ 3000 mg/m* iv. D;-Ds : [lacuna] 1 hour every 12 hours ®* daunorubicin 30-45 mg/m?/day bolus i.v, D;~Dq once/day 1.2.3. Treatment of adults aged 60 or older )

The flavonoids may be added to the consolidation chemotherapy protocols hereinafter: Co 3 i) according to R.O. Dilman et al. (Blood 1991; 78; : 2520-2526), Z.A. Arlin et al. (Leukemia 1990; 4: 5 10 177-183), P.H. Wiernik et al. (Blood 1992; 79: 313-319): es flavonoid 200-2000 mg/m°/day i.v. D;-Ds or 5-50 mg/kg/day 1 h infusion * cytarabine (Ara-C) 100-200 mg/m’ i.v. D;~Ds [lacuna] 24 hour continuous infusion e daunorubicin 30-45 mg/m"/day bolus] i.v. Dy, D2 or . oT e mitoxantrone 12 mg/m?/day bolus i.v. D1, Ds or : * idarubicin 13 mg/m?/dav bolus iv. Dy, Ds : ii) according to R.J. Mayer et al. (N. Engl. J. Med. 194; 331: 896-903):

dose lroure]| aays es flavonoid 200-2000 mg/m’/day | i.v. D;~Ds or 5-50 mg/kg/day 1 h infusion e cytarabine: 100 mg/m’ i.v. D;-Ds 24 hour continuous infusion (4 cycles) |. then e cytarabine 100 mg/m’ s.c. Dy, Ds, every 12 hours e daunorubicin 45 mg/m’/day bolus i.v. D; (4 cycles) 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): . This protocol comprises an autologous bone marrow transplant (carried out on day Dg): e flavonoid 200-2000 mg/m*/day i.v. D.7-D_; or 5-50 mg/kg/day 1 h infusion * busulphan 1 mg/kg gid oral |D.; to D. (16 doses in total) e etoposide 60 mg/kg/day i.v. D.3 hour infusion 10 or dose [route] days e flavonoid 200-2000 mg/m°/day | i.v. D.g-D_; or 5-50 mg/kg/day ~ J 1h infusion ’ e cyclophosphamide 50 mg/kg/day i1.v. | Ds to D, 1 hour infusion iv) in the event of HLA-compatible allogenic bone marrow transplant according to:

P.J. Tutscha et al. Blood 1987; 70: 1382-1388,

. F.R. Applebaum et al. Ann. Int. Med. 1984; 101: 581-588: dose [rouce| aays e flavonoid 200-2000 mg/m?/day i.v. D.,~D_, or 5-50 mg/kg/day 1 h infusion * Dbusulphan 1 mg/kg gid oral | D.; to D.; (16 doses in total) ®* cyclophosphamide 60 mg/kg/day i.v. | Ds to D, - 1 hour infusion 2°/ Adult chronic leukaemias ————— CTonic leukaemias 2.1 Chronic myeloid leukaemia -

In the myeloblastic phase, the flavonoids can be added to the HU-Mith treatment described by

C.A. Koller et al. (N. Engl. J. med. 1986; 315: 1433-1438): dose | route | gays |]

D,~Ds flavonoid 200-2000 mg/m°/day iv. Ds=Dia or 5-50 mg/kg/day Dis~Dia 1 h infusion D-2~Dag * mithramycin 25 pg/kg/day i.v. daily for 3 2-4 hour infusion weeks, then 3 times/week 2.2 Chronic lymphocytic leukaemia . 15 2.2.1 FCG-CLL protocol

The flavonoids can be added to the “pulsed chlorambucil” combinations as described by . E. Kimby et al. (Leuk. Lymphoma 1991; 5 (Suppl.) 93-96) and by FCGCLL (Blood 1990; 75: ’ 1422-1425);

» flavonoid 200-2000 mg/m?/day D1~Ds, D-g-Dy2, or 5-50 mg/kg/day i.v. D1s-D»s 1 h infusion e chlorambucil 0.1 mg/kg/day oral once/day or ¢ chlorambucil 0.4 mg/kg/day oral D; every 14 days and ¢ prednisone 75 mg/day oral D;-D; 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: 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+ flavonoid 200-2000 mg/m*/day D;-Ds or 5-50 mg/kg/day i.v. (once/month 1 h infusion for 6 to 12 cycles) e fludarabine 25-30 mg/m"/day iv. D;-Ds 30 minute infusion [every 4 weeks for 6 to 12 cycles] or e¢ cladribine 0.09 mg/kg/day i.v. D;-D, in a continuous infusion —— [1 cycle every 28 to 35 - ' ] days for 1 to 9 cycles (median: 4 cycles)] 3°/ Lymphoproliferative diseases ’ 3.1 Hodgkin’s disease

The flavonoids can be incorporated into the polychemotherapy protocols conventionally used for the treatment of Hodgkin’s lymphoma:

oa

K 3.1.1 AVDB protocol 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): 5 . dose [route] days e flavonoid 200-2000 mg/m*/day D;~Ds, or 5-50 mg/kg/day i.v. Di5—Dig 1 h infusion e doxorubicin (A) 25 mg/m? bolus e bleomycin (B) 10 U/m® bolus e vinblastine (V) 6 mg/m’ bolus * dacarbazine (D) 375 mg/m? bolus the therapy comprising 6 to 8 cycles, at a rate of 1 cycle every 28 days. 3.1.2 MOPP/ABVD protocol : 10 according to G. Bonnadonna et al. (Ann. Intern.

Med. 1986; 104: 739-746) and G.P. Canellos et al. (N. Engl. J. Med. 1993; 327: 1478-1484):

The MOPP protocol should be alternated with the 157 ABVD protocol (cf. 3.1.1) every 28 days, and the therapy comprises 6 cycles:

MOPP protocol: e flavonoid 200-2000 mg/m-/day D;-Dj, Dg-Dy; or 5-50 mg/kg/day i.v. and Dj;~Dy4 1 h infusion SE } e mechlorethamine (M) 6 mg/m” bolus e vincristine (0) 1.4 mg/m" bolus i.v. D;, Dg (no maximum) e procarbazine (P) 100 mg/m°/day oral | D-Day srednisons (7) 10 ma/n ay 3.1.3 Stanford V protocol according to N.L. Bartlett et al. (J. Clin.

Oncol. 1995; 13; 1080-1088):

dose |route| cays e flavonoid 200-2000 mg/m*/day D,-Ds, or 5-50 mg/kg/day i.v. Dg-J12 1h infusion Dis-Dis

D22.D2g e vinblastine 6 mg/m" bolus i.v. D;, Dis {4 mg/m® during cycle 3 if age 2 50) : e mechlorethamine 6 mg/m° bolus’ i.v. D: (M) e vincristine 1.4 mg/m® bolus i.v. D;, Dz» (maximum dose: 2 mg) (1 mg/m? during cycle 3 if age > 50) CL e prednisone 40 mg/m*/day once/week : {weeks 1-9) the therapy comprising 3 cycles, at a rate of 1 cycle every 28 days. 3.1.4 EVA protocol according to G.P. Canellos et al. (Proc. Am.

Soc. Clin. Oncol. 1991; 10: 273): e flavonoid 200-2000 mg/m’/day D,-Ds or 5-50 mg/kg/day | 1.v. 1 h infusion — e ctoposide (E) 100 mg/m’ oral D,, Da, Ds . - 2 hour infusion e vinblastine (V) 6 mg/m" bolus e doxorubicin (A) 50 mg/m” bolus the therapy comprising 6 cycles, at a rate of 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):

; | dose [route | days e flavonoid 200-2000 mg/m’/day D;~Ds or 5-50 mg/kg/day | i.v. 1 h infusion * bleomycin (B) 5 U/m?® bolus e lomustine (CCNU) 100 mg/m ¢ doxorubicin (A) 60 mg/m’ bolus e vinblastine (Ve) | 5s mg/m bolus | i.v. | Dp, the therapy comprising 8 cycles, at a rate of 1 cycle every 28 days. : 3.2 Non-Hodgkin’s lymphomas. 3.2.1. non-Hodgkin’s lymphomas with a” low degree of malignancy i) - CVP protocol - according to C.M. Bagley et al. (Ann. Intern.

Med. 1972; 76: 227-234) and C.S. Portlock et al. (Blood 1976; 47: 747-756) ~~ |e flavonoid 200-2000 mg/m>/day D,~Ds or 5-50 mg/kg/day i.v. 1 h infusion e cyclophosphamide (C) 300-400 mg/m"/day e vincristine (0) 1.4 mg/m bolus i.v. D, (max: 2 mg) es prednisone (P) 100 mg/m*/day

This cycle is repeated every 21 days until = maximum response ii) — I-COPA protocol : - according to RV Smalley et al/ (N. Eng. J. Med. . 1992; 327: 1336-1341)

Lo - 53 - ose | route | days e flavonoid 200-2000 mg/m’/day D;-Ds or 5-50 mg/kg/day i.v. 1 h infusion o cyclophosphamide (C) 600 mg/m’/day (max: 2 mg) e prednisone (P) : 100 mg/m’/day e doxorubicin (A) 50 mg/m’ bolus « interferon-alpha (I) - The therapy comprises 8 to 10 cycles, at a rate of one cycle every 28 days. iii) - fludarabine-Cda protocol To - according to P. Solol-Celigny et al. (Blood 1994; 84 (Supp. 1): 383a), H. Hoeschster et al.; (Blood 1994; 84 (Suppl. 1): 564a and A.C. Kay (J. Clin. Oncol. 1992; 10: 371-377) goss | zoute | days e flavonoid 200-2000 mg/m°/day D,-D; or 5-50 mg/kg/day i.v. 1 h infusion . le fludarabine 25 mg/m /day i.v. | Dy=Ds 0.5 hour infusion or e fludarabine 20 mg/m?/day i.v Dy~Ds and cyclophosphamide 600-1000 mg/m-/day i.v D, or cladribine 0.1 mg/m*/day i.v D,-Dy - - 24 hour infusion

For fludaribine, each cycle is repeated every 28 days; for cladribine, each cycle is repeated every 35 days. : 3.2.2. non-Hodgkin’s lymphomas with an intermediate ‘degree of malignancy

: i) —- CHOP or CNOP protocol - according to E.M. McKelvey et al. (Cancer 1976; 38: 1484-1493), J.0. Armitage et al. (J. Clin. Oncol. 1984; 2: 898-902), S. Paulovsky et al. (Ann. Oncol. 1992; 3: 205-2009) [dose route] days] e flavonoid 200-2000 mg/m?/day D;-Ds or 5-50 mg/kg/day i.v. 1 h infusion eo cyclophosphamide (C) 750 mg/m°/day iv. | Do e doxorubicin (H) 50 mg/m’ bolus e vincristine (0) 1.4 mg/m" bolus iv. D,; (max: 2 mg) e prednisone (P) 100 mg/m?/day _ oral | D;-Ds (in 1 dose/day) } for the CHOP protocol

Mitoxantrone (N) can be used to replace (CNOP : protocol) doxorubicin in patients over 60 years of age (dose: 12 mg/m? as an i.v. bolus on day D1 of each cycle).

The therapy using the CHOP or CNOP protocol : comprises 6 to 8 cycles, at a rate of 1 cycle every 21 days. ii) —- MACOP-B protocol - 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) oo dose route] days

D1-Ds, Dg=Dy2 e flavonoid 200-2000 mg/m?/day | i.v. { Dis~Ds, Dsg~Dy3 or 5-50 mg/kg/day D43=Dq1, Dg71-De1,

B 1 h infusion Dy1~Dig e methotrexate (M) 100 mg/m? bolus iv. Dg, Dig, Das . then 300 mg/m? ‘4 hour infusion e doxorubicin (A) 50 mg/m? bolus i.v. | Di,Dys,Dzg, Dys,

Ds, Dn e cyclophosphamide 350 mg/m’ bolus i.v. D1, Ds, Dag, Das, c (e) - Ds7, D1: e vincristine (0) 1.4 mg/m’ bolus iv. Dg, Daz, D3s, ax: 2 m (m 9) Dso, Des, Drg e prednisone (P) 75 mg/day oral Each day for } 12 weeks

This treatment protocol spreads out over 12 weeks and corresponds to 1 cycle. iii) - VACOP-B protocol - according J.M. Connors et al. (Proc. Am.

Soc. Clin. Oncol. 1990; 9: 254):

dose route] days

D1-Ds, Dg=D12 e flavonoid 200-2000 mg/m?/day | i.v. | Dys=Dzz2, D2s=D3e or 5-50 mg/kg/day D43—=D47, Ps7— Der, ) 1 h infusion D4y- Dis

D172, D13 e doxorubicin (A) 50 mg/m’ bolus . Dsv, D711 ¢ cyclophosphamide 350 mg/m?/day i.v. Dg, Das, Dig, (c) bolus Dso, Deas Drs

Dso, Dear Dig

Co e prednisone (P) 45 mg/m?/day oral.| 1l/day for 1 : week, then 4/day for the following 11 weeks : Each cycle lasting 12 weeks. iv) - m-BACOD/M-BACOD protocol - according to M.A. Shipp et al. (Ann. Int.

Med. 1986; 140: 757-765) and A.T. Skarin et al. (J. "Clin. Oncol. 1983; 1: 91-98)

L- ’ * - 57 - dose louse | days e flavonoid 200-2000 mg/m?/day| i.v. D;-Ds, Dg~D12 or 5-50 mg/kg/day Di5-Dis 1 h infusion e¢ methotrexate (m) 200 mg/m’ i.v. Dg, Dis 4 hour infusion or or (M) : 3000 mg/m’ i.v. Dis 4 hour infusion (6 doses in total) e doxorubicin (A) 45 mg/m’ bolus e cyclophosphamide 600 mg/m’ bolus iv. . Dy (C) . vincristine (0) | img/m bolus | iv. [Di o dexamethasone (D) 6 mg/m’/day

The therapy comprising 10 cycles, at a rate of 1 cycle every 21 days.

vr. “1 s, . : v) — ProMACE/CytaBOM protocol - according to D.L. Longo et al. (J. Clin.

Oncol. 1991; 9: 25-38): [dose | route | days e flavonoid 200-2000 mg/m’/day i.v. |D;-Ds,Dg-Diz or 5-50 mg/kg/day 1 h infusion e cyclophosphamide } 650 mg/m’ i.v. D, (C) 0.5 hour infusion e doxorubicin (A) 25 mg/m? bolus e etoposide 120 mg/m’ i.v. Dy : 1 hour infusion + bleomycin (B) 5 U/m* bolus iv. | © Dy 1.4 mg/m’ bolus | &.v. | Ds 120 mg/m’ bolus | i.v. | Ds e leucovorin 25 mg/m’ gid De (4 doses in total)

The therapy comprising 6 to 8 cycles, at a rate of 1 cycle every 14 days. 3.2.3. non-Hodgkin's lymphomas with a low or intermediate degree of malignancy i) ~ ESHAP rescue protocol ~ in the event of recurrence or in the event of failure of the first-line treatment, according to i

W.S. Velasquez et al. (J. Clin. Oncol. 1994; 12: =

J 1169-1176)

te Ea v cose | route | days e flavonoid 200-2000 mg/m?/day | i.v. | Di-Ds or 5-50 mg/kg/day 1 h infusion e etoposide (E) 40 mg/m? i.v. D;-~Dy 2 hour infusion e methylprednisolone (S) 500 mg/day iv. Di, Ds : 15 minute infusion e cytarabine (HA) 2000 mg/m’ i.v. Ds 3 hour infusion } e cisplatin (P) 25 mg/m’/day bolus | i.v. D,-Ds 24 hour continuous infusion

The treatment comprising 6 cycles, at a rate of 1 cycle every 28 days. Co } ii) - MINE rescue protocol - in the event of recurrence Or in the event 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 flavonoid 200-2000 mg/m? /day i.v. D1-Ds or 5-50 mg/kg/day 1 h infusion e ifosfamide (I) 1330 mg/m’ i.v. D;-Ds 1 hour infusion : eo mesna (M) 1330 mg/m’ i.v. | D;-Ds in the ifosfamide infusion, then 266 mg/m’ . polus 4 and 8 hours after _—

Lc each dose of ifosfamide 15% minute infusion e etoposide (E) 65 mg/m°/day i.v. D,-D; 1 hour infusion ’ This cycle being repeated every 21 days.

_ 3 > - 3.3 Non-Hodgkin’s lymphomas: Burkitt’s lymphoma, small cell lymphoma, lymphoblastic lymphoma. 3.3.1 Magrath protocol - The flavonoids may be combined with the

Magrath protocols according to the following schemes: i) - cycle 1 : - according to I.T. Magrath et al. (Blood 1984; 63: 1102-1111) dose | route | days e flavonoid 200-2000 mg/m°/day| i.v. D;-Ds or 5-50 mg/kg/day Dg-D12 1 h infusion e cytarabine 30 mg/m’ intra- Dy, Dz, D3, Ds thecal e cyclophosphamide 1200 mg/m’ bolus e methotrexate 12.5 mg/m? Intra- Dio (max 12.5 mg) thecal e methotrexate 300 mg/m°/day i.v. Dio~Di1 . 1 hour infusion then 60 mg/m’/h 41 hour infusion e leucovorin 15 mg/m° bolus i.v. To begin 42 (8 successive hours after doses) the start of the ad- ministration —— ! } of methotrexate ii) - cycles 2 to 15 ] 15 - according to I.T. Magrath et al. (1984) as well:

ns » ne » dose lroute | days e flavonoid 200-2000 mg/m*/ | i.v. D;-D; day D1o-D13 or 5-50 mg/kg/ - day 1 h infusion e cytarabine 45 mg/m’ Intra- Di, Da : . thecal | (cycles 2 and 3)

By (cycles 4 and 6) e cyclophosphamide | 1200 mg/m? bolus | i.v. D, (C) e vincristine 1.4 mg/m? bolus | i.v. D, (max 2 mg) Co } e methotrexate .12.5 mg/m? Intra- Ds, Dig (max 12.5 mg) thecal | (cycles 2 and 3)

Dio (cycles 4,5,6)

B e methotrexate 300 mg/m’ i.v. Dio, Duy 1. hour infusion (cycles 2 and 6 then Dis, Dis 60 mg/m’ (cycles 7-15) 41 hour continuous infusion oe leucovorin 15 mg/m” bolus i.v. To begin at the gid 42nd hour of the (8 consecutive treatment with doses) methotrexate the therapy comprising 14 cycles, at a rate of one cycle every 28 days. ~ 5 3.4 Waldenstrdém macroglobulinaemia 3.4.1 CVP protocol 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 flavonoid 200-2000 mg/m°/day | i.v. | Dy-Ds or 5-50 mg/kg/day 1 h infusion e cyclophosphamide (C) 300-400 mg/m?/day e vincristine (V) 1.4 mg/m‘/day bolus | i.v. | Di : (max: 2 mg) e¢ prednisone (P) 100 mg/m’/day the therapy being continued indefinitely (1 cycle every 21 days). 3.4.2 Fludarabine-CdA protocol - according to H.M. Kantarjian et al. (Blood 1990; 75: 1928-1931) and M.A. Dinopoulous et al. (Ann.

Intern. Med. 1993; 118: 195-198): [dose | route | days e flavonoid 200-2000 mg/m*/day | i.v. D;-Ds or 5-50 mg/kg/day 1 h infusion e fludarabine 25-30 mg/m’ i.v. D;-Ds 0.5 hour infusion or cose | route | days e flavonoid 200-2000 mg/m-/day | i.v. D;~D, or 5-50 mg/kg/day 1 h infusion — . e cladribine (CdA) 0.09 mg/m*/day i.v. D;-Ds continuous infusion the therapy comprising 6 to 12 cycles 28 days apart in the case of fludarabine, and 2 cycles 28 days ~ 15 apart also, in the case of cladribine. 3.5 Multiple myeloma 3.5.1 MP protocol according to R. Alexanian et al. (JAMA 1969; 50 208: 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 flavonoid - 200-2000 mg/m®/day | i.v. | D;-Ds or 5-50 mg/kg/day 1 h infusion e melphalan (M) E 0.25 mg/kg/day ¢ prednisone (P) 100 mg/day 3 or e flavonoid 200-2000 mg/m*/day | i.v. | Dy-Ds or 5-50 mg/kg/day 1 h infusion e melphalan (M) 9 mg/m*/day the therapy comprising at least 12 cycles, at a rate of 1 cycle every 4 to 6 weeks. 3.5.2 VAD protocol according to B. Barlogie et al. (N. Engl. J.

Med. 1984; 310: 1353-13506): e flavonoid 200-2000 mg/m*/day | i.v. D;-Ds or 5-50 mg/kg/day 1 h infusion : Tm ' } e vincristine (V) 0.4 mg/day i.v. D;-D, 24 hour continuous infusion e doxorubicin (A) 9 mg/m°/day i.v. D,-Dy 24 hour continuous ) infusion e dexamethasone (D) 40 mg/day i.v. | D;-Dy, Dg~Dia,

D;7-Dao

3.5.3 MP/interferon-a protocol according to O. Osterborg et al. (Blood 1993; 8l: 1428-1434): : dose [route] days ¢ flavonoid’ 200-2000 mg/m’/day iv. D;-Ds or 5-50 mg/kg/day 1 h infusion * melphalan (M) 0.25 mg/kg/day e interferon-alpha 7 MU/m"/day s.c. D;-Ds and

Daz=Dag } the therapy comprising the indefinite repetition of this cycle, at a rate of 1 cycle every 42 days. 3.5.4 VCAP or VBAP protocol according to S.E. Salmon et al. (J. Clin.

Oncol. 1983; 1: 453-461): : 10 protocol VCAP: e flavonoid 200~2000 mg/m*/day iv. D;~Ds or 5-50 mg/kg/day 1 h infusion * vincristine (V) 1 mg/m” bolus iv. D; (max: 1.5 mg) ¢ doxorubicin (A) 30 mg/m’ bolus {e cyclophosphamide 125 mg/m” oral D1-Dy, — = (C)

VBAP protocol: cyclophosphamide is replaced with carmustine (BCNU), the rest being identical: e carmustine 30 mg/m* i.v. D, 1 hour infusion oe { - 2 Pl “ a C. CHILDHOOD TUMOURS - Paediatric oncology i

The flavonoids can also be incorporated into polychemotherapy protocols for treatment of paediatric

S tumours, ‘in order to improve the antitumour effectiveness while at the same time reducing the severity of the side effects due to the action on the ; recruitment and mobilization of the clonogenic cells, and to the pgssibility of decreasing the active doses. 10 1°/ Ewing's sarcoma/primitive neuroectodermal tumour

The flavonoids can be introducéd into ‘the VCR-

Doxo-CY¥-Ifos-Mesna-E protocol (E.D. Bergert et al., J. 15 Clin. Oncol. 1990; 8: 1514-1524; W.H. Meyer et al., J.

Clin. Oncol. 1992; 10: 1737-1742): dose route| days e flavonoid 100-200 mg/m°/day D;-Ds and Day-Da, or 2-50 mg/kg/day iv. and Dy3-D;3 and 1 hour infusion Ds3~Dea e vincristine 2 mg/m‘ bolus i.v. | . Di, Ds, Dis, Dy (maximum dose = 2 mg) s doxorubicin 30 mg/m°/day i.v. D1-Ds, in a 24 hour infusion Dg3—Dys e cyclo- 2.2 g/m iv. D1, Das phosphamide in a 0.5 hour infusion » ifosfamide 1800 mg/m"/day i.v. Dp>~Dag in a 1 hour infusion Dg3~Dg7 —— e mesna 360 mg/m" i.v. |administered with , ] in a 15 minute cyclophosphamide ’ infusion, at a rate of and ifosfamide doses every 3 hours e etoposide 100 mg/m" i.v. D22=Dag : in a 1 hour infusion De3—Dg7

Pp the therapy comprises 6 to 10 of these cycles depending on the initial severity of the sarcoma and on the amplitude of the response. 2°/ Childhood acute lymphoblastic leukaemia . 2.1. Induction chemotherapy (days D;-D-30).

- » Md ’ t vo ' - 66 ~ gE The flavonoids can be added to the recommended protocols (P.S. 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; V.J. Land et al., : : J. Clin. Oncol. 1994; 12: 1939-1945): e flavonoid 100-200 mg/m?/day D;~Ds, or 2-50 mg/kg/day iv. Dg=Dy1, D1s—Dig 1 hour infusion Daa=Dsn * vincristine 1.5 mg/m® bolus i.v. D1, Ds, Dis, Das (maximum dose = 2 mg) e L-asparaginase 6000 IU/m’ “iim. 3 times/week for 3 weeks * prednisone 60 mg/m” oral D1 to Dy - as 3 doses/day * daunorubicin 25 mg/m’/day i.v. Dy, Dg, Dis and in a 15 minute infusion D-- : ® methotrexate depending on age intra- Dis, Dag thecal ® cytarabine depending on age intra- D, thecal depending on the result of the examination of the bone marrow, entry into the consolidation phase takes place on day Ds of the treatment protocol. 2.2 Consolidation/maintenance chemotherapy

The flavonoids can be introduced into 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.

Oncol. 1994; 12: 1939-1945) according to the following scheme:

. a x ‘ ) ry ’ - ’ dose | route | gays e flavonoid 100-200 mg/m?/day Dy-Ds, Dys-Dyg and or 2-50 mg/kg/day i.v. Dg4=Dgg, D101 D106, 1 hour infusion Dj0s=Di113, D12a—Diav eo cyclo- 1000 mg/m? i.v. D1, Dis, Dis phosphamide in a 0.5 hour infusion e L- 6000 U/m? i.m. 3 times/week between Dg; and Dis» es cytarabine 75 mg/m*/day i.v./szc. a sequence of 4 in a 15 minute or days starting on infusion D2, Dg, Dig, D23, D123, . D. e doxorubicin 25 mg/m’/day i.v. Deg, D1o1, Digs infusion purine treatment * methotrexate 20 mg/m*/day oral once/week between

D3s and Dj», and ’ between Dy; and the end of the treatment * prednisone 40 mg/m"/day oral 5 consecutive days (divided into 3 per month between : TT doses/day) Diss and the end of . the treatment eo vincristine 1.5 mg/m’ bolus i.v. Dogs D101, Dos, then (maximum dose once/month between = 2 mg) Digs and the end of the treatment _—— e methotrexate | depending on age intra-: | Di, Dg, Dis, Daa, Dias, Digg ' . thecal then once/3 months between D;,; and ’ : the end of the treatment

- [4 Md ’ ’ «3 + : 3°/ Childhood acute myeloid leukaemia

The flavonoids are added to the induction and consolidation/maintenance protocols according to the following schemes: i 3.1. Induction chemotherapy

According to .Y. Ravindranath et al., J. Clin.

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

Oncol. 1994; 12: 127-135; RJ Wells et al., J. Clin. i 10 Oncol. 1994; 12: 2367-2377:

I I e flavonoid 100-200 mg/m’/day - D;-Ds, Dyo-Di3 or 2-50 mg/kg/day i.v. ’ 1 hour infusion e cytarabine according to age intra- D; thecal * daunorubicin 20 mg/m" /day i.v. D;=Dy, Dyp-Dy;3 in a 24 hour infusion es cytarabine 200 mg/m" /day i.v. D1-Dy, D1o-Di3 in a 24 hour infusion * thioguanine 100 mg/m" /day oral D;—Dy, D1g-Di3 divided into 2 doses/day . |e etoposide 100 mg/m’/day i.v. D;-Dy, D1g=Di3 in a 24 hour infusion * dexamethasone 6 mg/m" i.v./oral| D;-D,, D;p-Dis divided into 3 doses/day this cycle being repeated from Dag. 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.

Oncol. 1994; 12: 2367-2377:

CL he ¥ v, » - 69 - dose | voure | days ® cytarabine according to age intra-~ D1, D2g, Dsg thecal : e flavonoid 100-200 mg/m’/day i.v. D;~Ds, Dg=Ds3 or 2-50 mg/kg/day D2g—D33, Deg=De1 1 hour infusion Dgoa—Da, e cytarabine 3000 mg/m? iv. D;-D; in a 3 hour infusion and Dg-Dy every 12 hours e l-asparaginase 6000 IU/m’ i.m. 3 hours after the cytarabine e Vincristine 1.5/m® bolus i.v. Dag, Dsg (maximum dose = 2 mg) Co - e Cyclo- 75 mg/m’/day i.v. Dzg=Day/ Dsg~Dssg phosphamide in a 0.5 hour infusion e Etoposide 100 mg/m’ /day i.v. Des, Das in a 1 hour infusion e Daunorubicin 30 mg/m" i.v. in a 15 minute infusion 4°/ Childhood Hodgkin’s disease

The flavonoids can be added to the MOPP-ABVD

S protocol according to 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):

FJ © - 70 = [dose route] aays e flavonoid 100-200 mg/m*/day i.v. D;-Ds or 2-50 mg/kg/day and Ds-Di2 1 hour infusion e mechlorethamine (M) 6 mg/m? bolus (maximum 2 mg) e procarbazine (P) 100 mg/m’/day

J prednisone (P) 40 mg/m /day oral D;~Dyg (divided into 3 doses/d) e doxorubicin (A) 25 mg/m’/day 1.v. | Dag, Dogs in a 15 minute infusion in a 15 minute infusion . e vinblastine (V) 6 mg/m" bolus | i.v. |". Dag, Du3 (maximum 2 mg) e dicarbazine (D) 375 mg/m’ iv. Daa, Das in a 15 minute infusion

This cycle should be repeated 6 times, at a rate of 1 cycle every 8 weeks, the therapy comprising 6 cycles.

If an autologous bone marrow transplant (autograft) is © prescribed, the CVB protocol described by R. Chopra et al. (Blood 1993; 81: 1137-1145), C. Wheeler et al. (J.

Clin. Oncol. 1990; 8: 648-656) and R.J Jones et al. (J. = clin. Oncol. 1990; 8: 527-537) may be used according to the following scheme (the allograft taking place on day

Do) : : ose route] ams e flavonoid 100-200 mg/m’/day D7, Da or 2-50 mg/kg/day i.v. 1 hour infusion e cyclophosphamide 1800 mg/m°/day iv. D-7, Dg ’ in 2 1 hour infusions D-s, Dy e carmustine (BCNU) 112 mg/m°/day i.v. D.7, Dg i in a 0.5 hour infusion D_e, D4 : e etoposide 500 mg/m>/day i.v. D.7,D-¢ in 2 1 hour infusions D_g, D-4 v ££ - iE 5°/ Childhood lymphoblasic lymphoma

The flavonoids may also be combined with induction chemotherapy (A.T. Meadows et al., 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 maintenance chemotherapy protocols: 5.1 Induction chemotherapy ase [rouse | cays e flavonoid : 100-200 mg/m-/day i.v. D,-Ds, or 2-50 mg/kg/day To D17=Daa, D2g—D2g 1 hour infusion : e cyclo- 1200 mg/m’ i.v. D, phosphamide in a 0.5 hour infusion e cytarabine according to age intra- Di thecal e vincristine 1.5 mg/m’ bolus i.v. (maximum 2 mg) divided into 3 doses/day "|e daunorubicin 60 mg/m’ i.v. D1s in a 15 minute infusion e IL -asparaginase 6000 U/m’/day im Dy7-Das in a 15 minute infusion 3 times/week e Methotrexate according to age intra- Diq, Don thecal 5.2 Maintenance chemotherapy oT according to the following scheme:

] ¢* Ll “ 3 | orn - 72 - i ! cose | rouwe | days e Flavonoid 100-200 mg/m*/day iv. D;~Ds, Dis=Dso, or 2-50 mg/kg/day Dayg—-Ds, 1 hour infusion * Cyclophosphamide 1000 mg/m? i.v. D, in a 0.5 hour infusion : ¢ Vincristine 1.5 mg/m*> bolus oral Dy, Ds, (maximum 2 mg) (of cycles 2 to 10) e Methotrexate 300 mg/m°/day i.v. Dis (60% in a 15 minute infusion and 40% in a 4 hour infusion) e Daunorubicin 30 mg/m’ i.v. Dag in a 0.5 hour infusion ® Methotrexate according to age intra- Di, Dg, Dis thecal | (cycle 1) then once/month (cycles 2 to 10) the therapy comprising 10 cycles 8 6°/ Paediatric neuroblastoma a

The recommended Doxo-E-Cy-Pt polychemotherapy protocol is adapted from R.P. Castleberry et al. (J.

Clin. Oncol. 1992; 10: 1299-1304), A. Garaventa et al. (J. Clin. Oncol. 1993; 11: 1770-1779) and D.C. West et mT al. (J. Clin. Oncol. 1992; 11: 84-90):

: & 1 LA -~ dose | rome | ape * flavonoid 100-200 mg/m®/day iv. .D;~Ds, ! or 2-50 mg/kg/day D2g=Dss, i 1 hour infusion Dsg=Dgs * doxorubicin 25 mg/m?’/day i.v. Da, Dag, Deg in a 15 minute infusion * etoposide ~ 100 mg/m? oral/ Dz, Ds, D3g, D3, in a 1 hour infusion naso- Dsg, De; . gastric * cyclophos- 1000 mg/m’ iv. Ds, D;, Day, Daz, phamide in a 0.5 hour infusion Dsg, Deo * cisplatin 60 mg/m’ i.v. D1, Dog, Dsg in a 6 hour infusion .

The evaluation of the therapeutic response is carried out after 9 weeks in order to decide upon the approach: surgical resection, radiotherapy or new chemotherapy. 7°/ Paediatric osteosarcoma

The flavonoids can be added to the Doxo-Pt-Mtx-~Lcv protocol as is 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):

I RF

» flavonoid 100-200 mg/m*/day i.v. D1=Ds, D21=Dag, — or 2-50 mg/kg/day Dag~D3y } 1 hour infusion : e doxorubicin 25 mg/m°/day i.v. D;-D;3 in a 24 hour infusion e cisplatin 120 mg/m* i.v. D; in a 6 hour infusion * methotrexate 12 mg/m’/day i.v. Day, Dag : in a 1 hour infusion e leucovorin 100 mg/m’ oral Daa, Dag i every 6 hours

E o ’, “~ . i

EC - 74 - 8°/ Childhood rhabdomyosarcoma

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) can include the i.v. : infusion of the flavonoids according to the 1 following scheme: ! dose Tome | ape. |] e flavonoid 100-200 mg/m" /day i.v. D;~Ds, Dg-D;2, : or 2-50 mg/kg/day Da2~D37, Dy3-Dyq ; 1 hour infusion : e vincristine 1.5 mg/m*/day Liv. Di, Dg, D1s, Daz, bolus maximum 2 mg i Dag, D3g, Dy3,

Dsg, Dsy * dactinomycin 0.015 mg/kg bolus iv. D;-Ds, (max daily dose: 0.5 mg) D22=Ds7, Dy3-Dys e cyclo- 2.2 mg/m’ iv. Dy, D22, Dy3 phosphamide in a 1 hour infusion * mesna 360 mg/m" i.v. Dy, D22, Dy3 in a 1 hour infusion every 3 hours for 5 doses

At the end of the 9th week of treatment, the : 10 effectiveness should be evaluated to decide upon the Subsequent measures (surgery, radiotherapy, continuation of the chemotherapy) . 9°/ Wilms’ tumour in children

In the Vcr-Dact protocol as is described by oT

GJ D’Angio et al. (Cancer, 1989; 64: 349-360) and : DM Green et al. (J. Clin. Oncol. 1993; 11: 91-95) : dose route days e flavonoid 100-200 mg/m" /day i.v. Dy1-De, Dg—D;> - or 2-50 mg/kg/day then each week 1 hour infusion ¢ vincristine 2 mg/m’ bolus i.v. Dy (max dose: 2 mg) then each week e dactinomycin 0.045 mg/kg bolus i.v. Di, then every (P < 30 kg) 3 weeks 1.35 mg/m* (P>30 kg) (max dose: 3 mg)

\ _ - . -> 5 © HY co - 75 - - This protocol being started after surgical resection.

In the event of autologous bone marrow transplant (autograft) according to A. Garaventar et al. (Med. Pediatr. Oncol. 1994; 22: 11-14), the E-

Thio-Cy protocol may be modified as follows: dose [route] aays * ‘flavonoid 100-200 mg/m’/day iv. D.g-D_, or 2-50 mg/kg/day 1 hour infusion * etoposide 1800 mg/m’ i.v. 24 hour infusion BE * thiotepa 300 mg/m’/day i.v. D.;,D.s,D_s in a 2 hour infusion * cyclophosphamide 50 mg/kg/day i.v. D.y,D3,D.5,D, in a 1 hour infusion the bone marrow transplant taking place on Do.

Claims (10)

AO Co WO 00/03706 PCT/FR99/01714 76 CLAIMS

1. Composition which has activity on the proliferation of clonogenic cells in tumours and which comprises a therapeutically effective amount of a flavonoid. -

2. Composition which has activity on the proliferation of clonogenic cells in tumours and which comprises a therapeutically effective amount of a compound chosen from the compounds of formula: R1 } »e SN Re Co : (KL (0) Rs Rs R4 O : this being a formula in which: - Ris Rz2, Rs and Rg are chosen, independently of each other, from H, OH, a C;-C; alkoxy group and an -~0OCOR; group, Ry; being a C;-C4 alkyl group, at least one of the substituents R;, R;, R; or Ry; being other than H, and R, and R;3 possibly forming together a methylenedioxy group, - Rs is chosen from H, OH, a Ci;-C, alkoxy group and an 0O-glycosyl group, - Rg is chosen from a cyclohexyl group, a phenyl group and a phenyl group substituted 1 to 3 times with groups — py chosen from H, OH and a C;-C4; alkoxy group, 3 - and "TTT CC designates either a double bond or a single bond.

3. Composition according to Claim 2, in which the flavonoid is a flavone.

i 4. Composition according to Claim 1, in which the flavonoid is quercetin.

5. Use of a flavonecid for manufacturing a medicine intended to interfere with the generation of clonogenic cells in tumours during treatment of these tumours with at least one cytotoxic agent.

PO SR

6. Use of a compound chosen from the compounds of formula R4 peso : (KL 0) Ra Rs R, 4 this being a formula in which: 3S = Ri, Ry, Ry and Ry are chosen, independently of each other, from H, OH, a Ci-Cq alkoxy group and an ~QCOR~y group, Ry being a C;~C, alkyl group, at least one of the substituents R;, R,, R3 or Ry being other than H, and R; and Ry possibly forming together -a- methylenedioxy group, - Rs 1s chosen from H, OH, a C;-C; alkoxy group and an O-glycosyl group, - R¢ 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 Ci1-Cy alkoxy group, - and TTTUTTT designates either a double bond or a single bond, for manufacturing a medicine intended to interfere with the generation of clonegenic cells in tumours during treatment of these tumours with at least one cytotoxic agent.

7. Use according to Claim 6, in which the flavonoid is a flavone.

8. Use according to Claim 5, in which the compound oo of formula I is quercetin.

i 9. Method for chemotherapeutic treatment of a tumour, in a patient, with at ‘least one cytotoxic agent, which comprises the administration, during the treatment with the cytotoxic agent, of a therapeutically effective amount of a flavonoid.

10. Method according to Claim 9, in which the flavonoid is administered at the start of the chemotherapeutic treatment and at the start of each cycle of chemotherapeutic treatment.