AU7304096A - Novel amine derivatives of epipodophyllotoxin 2", 3"-dideoxyglycosides, preparation method therefor and use thereof as a drug and for treating cancer - Google Patents

Description

WO 97/13776 PCT/FR96/01588 Novel amine derivatives of epipodophyllotoxin 2",3" -dideoxyglycosides, preparation method therefor and use thereof as a drug and for anticancer treatments The present invention relates to novel amino 5 derivatives of epipodophyllotoxin 2",3"-dideoxyglyco sides, the method for preparing them, their use as a drug and their use for anticancer treatments. The class of epipodophylloids, having the basic podophyllotoxin backbone, is among the derivatives 10 derived from natural lignans. Among them are the semi synthetic derivatives such as Etoposide or Teniposide which are customarily used in the preparation of drugs for the treatment of cancer. They are considered as major products in this field. 15 Etoposide has antitumor properties and makes it possible to treat, in particular, small cell lung cancer and cancer of the testicles. The disadvantage of these products is their lack of solubility in water and they consequently encounter 20 difficulties in terms of formulation and administration. The object of the present invention is to show that 4'-demethylepipodophyllotoxin derivatives posses sing, at the 4-position, a substitution of the 2"-deoxy glycoside structure, makes it possible, by the incor 25 poration of one or more nitrogens, to form compounds whose addition salts possess aqueous solubility making it possible to address the problem and show the desired anticancer activity. Patent EP-0,196,618 describes water-soluble 30 derivatives of 4'-demethylepipodophyllotoxin of formula: - 2 R~O oo 00 where Ri = Me, X 1 = NH 2 , NMe 2 , X 2 = OH Patent EP-0,415,453 mentions $-D-altroside derivatives of 4'-demethylepipodophyllotoxin of formula R, O, 0I 0 0. / -OO OH where R, = Me R 2 , R 3 = OH and NH 2 or F and NH 2 as well as 5 JP 0,161,423. Other publications mention similar derivatives (Carbohydr. Res. 1990, 206, 219; Chem. Pharm. Bull. 1986, 34, 3733; Chem. Pharm. Bull 1986, 34, 3741; Chem. Lett. 1987, 799). 10 The fact that a glycoside having a 2"-deoxy position is used is quite special. It makes it possible to obtain compounds which are more lipophilic than hydroxylated analogs and, consequently, to have a broadened antitumor activity spectrum. This allows them 15 to have better membrane penetration and to be able more easily to reach the biological target such as, for example, solid tumors which are not highly irrigated. In addition, the advantage of having an amino functional - 3 group, for example, in the 3"-position confers a possi bility of salification and therefore of water-solubility which is sufficient for a better formulation and a better administration. 5 The present invention therefore relates to a compound of general formula I Y X 0 0 MOO MeOr OMe OH in which the group in 3" N(RlR 2 ) is in the P-position (2-deoxy-D-Arabino series) or a-position (2-deoxy-D-ribo series) in relation to the ring, R, and R 2 which are 10 identical or different, represent a hydrogen atom, a C 1 to C 6 alkyl group, capable of forming a ring, it being possible for this ring to contain a heteroatom such as oxygen or nitrogen, a C 1 to C 6 aminoalkyl group or a cyanomethyl group. 15 X and Y may be identical or different and repre sent OH, CH 3 , CH 2

-NH

2 , X and Y may also be linked and may constitute a ring, such as for example a 2-methyl-1,3 dioxane, thus forming a bicyclic glycoside backbone of the 4, 6-ethylidene-3-amino-2, 3-dideoxy-$-D-arabino- or 20 -ribo-hexo pyranoside. It also relates to their addition salts with inorganic or organic acids salifying the nitrogen atom(s), in particular the hydrochlorides. Advantageously, the NR 1

R

2 group is an NH 2 or 25 N(CH 3

)

2 group. The NR 1

R

2 group may also be an amino group sub stituted once or twice with a methyl, CH 2 CN, CH 2

-CH

2

-NH

2

,

- 4 to form a ring such as morpholine. Advantageously, the compounds of general formula I are chosen with a glycoside for which X and Y form a ring with an OCH(CH 3

)OCH

2 linkage, such as 4,6-ethyli 5 dene-3-amino-2, 3-dideoxy-$-D-arabino-hexopyranoside or 4, 6-ethylidene-3-amino-2, 3-dideoxy- -D-ribo-hexopyrano side. In particular, the compounds according to the invention are selected from the following compounds: 10 - 4'-demethyl-4-0(3-amino-4,6-ethylidene-2,3-dideoxy $-D-arabinohexopyranosyl) epipodophyllotoxin, - 4'-demethyl-4-0 (3-amino-4,6-ethylidene-2,3-dideoxy S-D-ribohexopyranosyl) epipodophyllotoxin, - 4'-demethyl-4-0 (3-dimethylamino-4,6-ethylidene-2,3 15 dideoxy-$-D-arabinohexopyranosyl) epipodophyllotoxin, - 4' -demethyl-4-0 (3-dimethylamino-4, 6-ethylidene-2, 3 dideoxy-$-D-ribohexopyranosyl) epipodophyllotoxin, - 4'-demethyl-4-0 (3-cyanomethylamino-4, 6-ethylidene 2,3-dideoxy-$-D-ribohexopyranosyl)epipodophyllo 20 toxin, - 4'-demethyl-4-0(3-(N-morpholino)-4,6-ethylidene-2,3 dideoxy-$-D-ribohexopyranosyl) epipodophyllotoxin, - 4' -demethyl-4-0 [3 (2-aminoethylamino) -4, 6-ethylidene 2,3-dideoxy- -D-ribohexopyranosyl)]epipodophyllo 25 toxin, - 4' -demethyl-4-0 (3-amino-2, 3, 6-trideoxy-$-D-ribohexo pyranosyl) epipodophyllotoxin, - 4'-demethyl-4-0(3,6-diamino-2,3,6-trideoxy-$-D ribohexopyranosyl)epipodophyllotoxin. 30 The present invention also relates to the pharma ceutical compositions comprising at least one compound of general formula I according to the invention and an appropriate excipient. The pharmaceutical compositions may be provided 35 in an appropriate manner by administration by the injectable route or by the oral route in the form of a capsule, gelatin capsules or tablets at the dosage of 1 to 200 mg/m 2 by the injectable route and of 5 to 500 mg/m 2 by the oral route per period of 24 h.

- 5 These derivatives can thus be administered in human clinical medicine to treat various forms of cancer such as small cell lung cancer, cancer of the testicles, embryonic tumors, neuroblastomas, kidney cancer, 5 Hodgkin's and non-Hodgkin's lymphomas, acute leukemias, colorectal cancers, melanomas, placental choriocarcinomas and mammary adenocarcinomas. The present invention also relates to the methods for preparing the compounds of formula I as well as their 10 addition salts with pharmaceutically acceptable inorganic or organic acids. The present invention therefore relates to the methods for preparing the compounds of general formula I according to the invention, in which a compound of 15 formula III or IV or V V its Iv is reacted with 4'-demethyl-4'-benzyloxycarbonylepipodo phyllotoxin with BF 3 etherate, or trimethylsilyl tri fluoromethanesulforate in an inert solvent at low temperature; 20 in formula III and IV, the substituent at the 3-position may be a or 0, NR 1

R

2 may be an amino protected with a group Z, in formula V, P represents an alcohol-protecting group and the resulting products of this condensation are 25 deprotected and hydrogenated to give the compounds of formula I, the primary amines in the 3-position of the glycosyl are methylated with formalin and sodium cyano borohydride. 30 The intermediate of formula IV is prepared by reacting a mixture of diacetoxyazidoglycoside VI - 6 ACO AcO OH vt with tert-butyldimethylsilyl chloride in the presence of imidazole, in that the products resulting from this reaction are separated, in that each of these products are deacetylated, cyclized to 4,6-ethylidene with 5 acetaldehyde acetal in a catalytic acid medium. Other characteristics of the process according to the invention will emerge in the light of the following description, in particular of the method of synthesis reported in Scheme 1.

7 SCHEME 1 Ac Ac 1) H120 8o-C Ac Ac 2) NaN3I/AcOR A co O NJ 2 TBDDMSiCI / Imidazole CH2CIZ AcO AcO 01i +AcO osi--- Ac

N

3

N

3 4 3 OH S paraton 0 i HO O N MCOH 5 McONa R.T. Ac. Ciumphorsulfoniquc ou APTS CH3CN R.T. 00 HI2C/Pd 10% N. "-" AcOEt.NEt3 6 0. ZCIINE. CH2CI2 ZNH o cNsr 0 11 2 N S1) N~u4 F, TlIF7 C14202 2) 13F3EL2O DMEPT 4-OZ -8 H2 C/Pd 10% 0 0 AcOEtNEt3O ZN 1 R.T. DMEF 10 DMEPT 4'-OZ 4'-0l1 1) HCHO NaBlH3CN CH2C2 0 R.T. 0 0 2)14C1: MCOH Me 2 N 2C2' 4-Oli MeOH OR McONa 0 R.T. 0 s CN 3CH(OEt)2 N, 13 N 3 12 CII13CN N31 Ac.camphosulfonique TMSOTf CH2C2 Tamis 4* -4O* DM E PT4-OV O 0 0 0 BF3-Et20 CH2CI2 N, 14 DMEPT -150 4.-OZ DMEPT 4'-OZ H2 C/Pd 10% EtOH-AcOEt 00 0 0 0 0 0 O O O /DMF 1 N DMEPT DMEPT 4'-H 4'-OH 41- N NH DM IEPT 'H O 17 0 02C212 16 4'-OH I C1+2LH2NHZ DMF HCHO NEt3 NaBH3CN CNt3CN R.T. 0 0 H2 C/Pd 0 0 AcOEt-EtOH 0 Me2N DMEOH 0 - 02 0 4'-O H DNHEPT HN DEPT 19 18 - 9 It is carried out according to a methodology which has been described (J. C. Florent and C. Monneret, J. Chem. Soc. Chem. Comm. 1987, 1171 and B. Abbaci, J. C. Florent and C. Monneret, Bull. Soc. Chim. Fr. 1989, 5 667) from glucal 1. The azide ion is condensed to give a glycoside intermediate 2 whose anomeric OH is protected with a silyl group exclusively in the 0-position accord ing to the technique which has been described (C. Kolar and G. Kneissl Angew. Chem. Int. Ed 29, 809 (1990) ) to 10 give the mixture of 2 epimeric azides: 3 and 4 which can be chromatographically separated at this stage. By basic deacetylation in the presence of sodium methoxide, the diols at the 4,6-position are obtained: compounds 5 and 12. The $ azide diol compound 5 is cyclized to ethylidene 15 conventionally with the aid of acetaldehyde acetal using acid catalysis to give the compound 6 whose azide is reduced to an amine 7 so as to be protected with a benzyloxycarbonyl group (Z) in compound 8, which is necessary to carry out the coupling with demethylepipodo 20 phyllotoxin, itself protected on its phenol in 4' with a benzyloxycarbonyl group, this intermediate will be called DMEPT4'-OZ. This coupling is carried out, in a first instance, .by cleaving the protective silyl with F~ ions, followed by treatment with BF 3 etherate at low tempera 25 ture in the same medium. Deprotection of the Z groups by hydrogenolysis provides compound 10 of general formula I

(NR

1

R

2 = PNH 2 ; XY = OCH(CH 3

)OCH

2 ) . Compound 11 of general formula I (NR 1

R

2 = PNMe 2 ; XY = OCH(CH 3

)OCH

2 ) is obtained by methylation of the preceding primary amine by the 30 action of formalin and sodium cyanoborohydride. The a azide glycoside intermediate 12 follows the same sequence of reactions as its epimer and provides, in an identical manner, the derivative from the coupling with DMEPT4'OZ starting with the a 3" azide 13; in this 35 case, the coupling is carried out more easily by two techniques. The first technique consists in treating the ribohexopyranoside derivative 13 with trimethylsilyl trifluoromethanesulfonate (TMSOTf) at -40 0 C in CH 2 Cl 2

-

- 10 The second technique consists in using BF 3 etherate in

CH

2 Cl 2 at -15 0 C. The intermediate obtained 14 is then catalytically reduced to give 15 corresponding to the general formula I where NR 1

R

2 = OCNH 2 and 5 XY = OCH(CH 3

)OCH

2 . The same methylation as for the PNH 2 compound 10, using formalin and cyanoborohydride, leads to the corresponding dimethylamino derivative: general formula I (NR 1

R

2 = aNMe 2 ; XY = OCH(CH 3

)OCH

2 ). Using the primary amine 15, it is possible to alkylate the nitrogen 10 with a halogenated derivative such as iodoacetonitrile under weakly basic conditions with triethylamine in DMF and give the derivative 16 of general formula I where

NR

1

R

2 = cNHCH 2 CN; XY = OCH(CH 3

)OCH

2 . In the same manner, the derivative 17 is obtained by forming the morpholine 15 ring by cyclization of the diiodinated ether on the same primary amine intermediate 15. The same alkylation with the iodoethylamine protected by a group Z on compound 15 gives the derivative 18 which is catalytically reduced to 19, a diamino derivative corresponding to the general 20 formula I with NRjR 2 = aNHCH 2

CH

2

NH

2 ; XY = OCH(CH 3

)OCH

2 . The compounds of formula I where XY does not form a ring are obtained in the following manner represented in Scheme 2: - 11 SCHEME 2 OTs HO ~ ~ TsCI / Pyridine HO Nai/ Acetone HO OS O I + a ceoH N 12 CH2Cl2 N 2 N 22 ICICH2COCI Pyridine CH2CI2

N

3 IDMEPT 4'-OZ Oo NaN3 Et2O-BF3 O NcOOF CIAc 0 -15*C C cOOSi N DMEFT N DMEPT N3 23 N 3 2 7 4 ' O ZN 3 2 4 4 '- O Z I Amberite 410Amberlite41 CH2CI2-MeOH CH2CK2IMeOH

N

3 H 3 H2 CIPd o HO HO AcOEt HO o HOH 0 NEt3 N DMEPT NH DMEPT N, DMEPT 4'-OZ P4'-OH 25 4--OZ H2 C/Pd 26 AcOEt-EtOH NEt3 H2N 0 HO o NH2 DMEPT 4'-OH 29 The silylated a azide 12 is selectively tosylated with tosyl chloride in pyridine to 21, the tosylate of the primary alcohol is exchanged to an iodo derivative 22. At 5 this stage, the secondary alcohol in the 4-position is protected by a chloroacetate to give the functionalized 2-deoxysugar 23 ready to be condensed with DMEPT 4'-OZ under the customary conditions with BF 3 etherate in methylene chloride at low temperature. The intermediate 10 24 obtained makes it possible, by passing over the amberlite IRA410 resin treated in a basic medium, to deprotect the chloroacetate group into compound 25 and final catalytic hydrogenation makes it possible, in one step, to reduce the azide functional group to an a 3" 15 amino, to deprotect the Z functional group at the 4' position and to reduce the 6" carbon to methyl in order to provide the derivative 26 corresponding to the general formula I where NR 1

R

2 = 3" a NH 2 ; X = OH; Y = CH 3 . The intermediate 24 obtained above can react with the azide 20 ion in DMF at room temperature to give 3"aN 3 , 6"-N 3 - 12 containing an azidoacetate at the 4"-position 27, which can be cut into a 4" alcohol by a treatment similar to the preceding one: passage over Amberlite IRA 410 exchange resin to give the diazidoalcohol 28. The final 5 catalytic reduction step makes it possible to obtain the diamino derivative 29 corresponding to the general formula I (where NRjR 2 = 3"cNH 2 ; X = OH; Y = CH 2

NH

2 ). The salts formed from the nitrogenous compounds are, for example, hydrochlorides and are conventionally formed by 10 treating a methanolic solution of the nitrogenous compound with a stoichiometric, in relation to the sites to be salified, solution of hydrochloric methanol previously prepared. The crystallized hydrochloride can be optionally obtained by precipitation in the reaction 15 medium by addition of ethyl ether. The following examples are given as a guide and to illustrate the various steps of the synthesis, but with no limitation being implied. EXAMPLE 1 20 General formula I: NRjR 2 = ONH 2 ; XY = OCH(CH 3

)OCH

2 4'-Demethyl-4-0(3-amino-2,3-dideoxy-4,6-ethylidene-$-D arabinohexopyranosyl) epipodophyllotoxin (compound 10) Step 1 A solution of tri-O-acetyl-D-glucal 1 (50 g; 25 183 mmol) in water (400 ml) is heated for 3 h at 80 0 C. The reaction medium is then cooled to 20 0 C before addi tion of sodium azide (17.9 g; 275 mmol) and acetic acid (38 ml; 600 mmol) . After stirring at room temperature for 24 h, the medium is neutralized with NaHCO 3 (salt). The 30 phase is extracted with ethyl acetate (3 x 500 ml). The organic phases are combined, dried over MgSO 4 and then concentrated under reduced pressure. This gives 51 g of crude product 2 immediately treated as follows. Characteristics: TLC: cyclohexane/AcOEt: 1/1; Rf = 0.43 35 ClQH 1 5

N

3 0 6 M = 273 Step 2 Tert-butyldimethylsilyl-3-azido-2,3-dideoxy- 4 ,6-di-0 acetyl- -D-arabinohexopyranoside 3 and tert-butyldi methylsilyl-3-azido-2, 3-dideoxy-4, 6-di-O-acetyl- -D- - 13 ribohexopyranoside 4 6 g of imidazole (87.8 mmol) and 13.24 g of tert butyldimethylsilyl chloride (87.8 mmol) are added succes sively, under argon, to a solution of the crude mixture 5 2 (16.1 g; 58 mmol) obtained in step 1 in anhydrous dichloromethane (200 ml) previously cooled to OC. After stirring for 15 min at 0 0 C and 19 h at 20 0 C, the reaction medium is poured into 500 ml H 2 0. The aqueous phase is extracted with CH 2 Cl 2 (200 ml) and then after drying over 10 MgSO 4 , the organic phase is concentrated under reduced pressure and the residue (18.7 g) is chromatographed on a silica gel (cyclohexane/AcOEt: 9/1). 10.7 g of 3 (syrup; 48%) and 4.7 g of 4 (syrup; 21%) are thus isolated; whereas intermediate fractions contain a 15 mixture of 3 and 4 (3.3 g; 15%). Characteristics: 3 TLC: cyclohexane/AcOEt: 4/1 Rf = 0.45 [ = -00 (c = 1.4; CHCl 3 ) MS: m/z 405 (M+NH 4 ) 20 C 1 6

H

2 9

N

3 0 6 SiM = 387 4 Rf = 0.

5 2 {X]20 = +100 (c = 1; CHCl 3 ) MS: m/z 405 (M+NH 4 )+

C

1 6

H

2 9

N

3 0 6 SiM = 387 25 1 H NMR 300 MHz CDCl 3 0 Derivative 3: 0.13 (3H,s, SjCH 3 ); 0.14 (3H, s SjCH3); 0.92 (9H, s, tBu); 1.72 (1H, m, J 2 a-1 = 9.5 Hz, J2a-2e 12.5 Hz, J 2 a- 3 12.5 Hz, H2a) 30 2.05 (3H, s, COCH 3 ); 2.15 (3H, s, COCH 3 ); 2.25 (1H, ddd, J 2 e-1 = 1.5 Hz, J2e-2a = 12.5 Hz, J 2 e-3 = 4.5 Hz, H2e); 3.55 - 3.62 (2 H, m, H 3 and H 5 ); 4.10 (1H, dd, 35 J6-5 = 2.5 Hz, H 6 ); 4.20 (1H, dd, J 6

-

5 = 6 Hz, J6-6' = 12 Hz,

H

6 ,); 4.86 (1H, t, J = 9.5 Hz,

H

4 ); 4.86 (1H, dd, J1- 2 a 9.5 Hz, J1- 2 e = 1.5 Hz, H) .

- 14 Derivative 4: 0.1 (6H, s, Si(CH 3

)

2 ); 0.88 (9H, s, tBu); 1.64 (1H, ddd, J2a-2e = 14 Hz, J 2 a-1 = 8.5 Hz, J 2 a- 3 = 3.5 Hz, H2a); 2.03 (1H, ddd, 5 J2e-2a = 14 Hz, J 2 e-1 = 2 Hz, J2e-3 = 4 Hz, H2e); 2.05 (3H, s,

COCH

3 ); 2.13 (3H, s, COCH 3 ); 4.05 - 4.12 (1H, m, H 5 ); 4.17 - 4.22 (3H, M, H 3 , H 6 and 10

H

6 ,); 4.89 (1H, dd, J 4 -5 9.5 Hz, J 4

-

3 = 3.5 Hz, H 4 ); 5.02 (1H, dd, Ji- 2 a = 8.5 Hz, J1- 2 e 2 Hz, H 1 ). Step 3 15 Tert-butyldimethylsilyl-3-azido-2, 3-dideoxy- -D-arabino hexopyranoside 5 A solution of 1 M sodium methoxide (1.9 ml) is added, under argon, to a solution of the derivative 3 (3 g; 7.7 mmol) obtained in step 2 in anhydrous methanol 20 (40 ml). After reacting for 1 h 30 min at 20 0 C, the reaction medium is adjusted to pH = 7 by addition of H' resin (amberlite IRC 50 S). The reaction mixture is filtered and the filtrate concentrated under reduced pressure, giving 2.27 g of 5 (97%). 25 Characteristics: TLC: cyclohexane/AcOEt: 2/1; Rf = 0.36 = -260 (c = 1; CHCl 3 ) MS: m/z 304 (M + H)+321 (M + NHg)+ m.p. = 70-72 0 C

C

12

H

2 5

N

3 0 4 SiM = 303 30 Step 4 Tert-butyldimethylsilyl-3-azido-2,3-dideoxy-4,6-0 ethylidene-$-D-arabinohexopyranoside (6) 0.94 ml (6.6 mmol) of acetaldehyde diethyl acetal and then 15 mg of para-toluenesulfonic acid (0.08 mmol) 35 are added to a solution of 5 (0.20 g; 0.6 mmol) obtained in step 3 in 5 ml of acetonitrile. The reaction medium is stirred at room temperature for 1 h and then diluted with ethyl acetate (20 ml) before washing with a sodium hydrogen carbonate solution (pH = 9) (20 ml) and then - 15 with water (20 ml). The organic phase is dried over MgSO 4 and then concentrated under reduced pressure to give 0.25 g of crude product. Silica gel purification (cyclo hexane/AcOEt: 95/5) makes it possible to isolate 0.19 g 5 of pure 6 (86%). Characteristics: TLC: cyclohexane/AcOEt: 7/3; Rf = 0.89 [a] 20 = -19o (c 1.1, CHCl 3 ) MS: m/z 347 (M + NH 4 )*

C

14

H

27

N

3 0 4 SiM = 329 10 C H N Calculated 51.04 8.26 12.75 Found 51.64 8.43 12.51 Step 5 Tert-butyldimethylsilyl-3-amino-2,3-dideoxy-4,6-0-ethyli 15 dene--D-arabinohexopyranoside (7) 50 pl of triethylamine and then 0.5 g of 10% palladium on carbon are added to a solution of 6 (2 g; 6 mmol) obtained in step 4 in 30 ml of ethyl acetate. The reaction medium is placed under a hydrogen atmosphere 20 (atmospheric pressure). After stirring for 6 h at room temperature, the catalyst is removed by filtration and the organic phase concentrated under reduced pressure to give 1.82 g of pure 7 (98%). Characteristics: TLC: cyclohexane/AcOEt: 1/1; Rf= 0.23 *25 [a]20 = -28* (c = 1.3; CHCl 3 ) MS: m/z 304 (M + H)*

C

14

H

29

NO

4 SiM = 303 Step 6 Tert-butyldimethylsilyl-3-aminobenzyloxycarbonyl-2,3 30 dideoxy-4, 6-O-ethylidene-#-D-arabinohexopyranoside (8) Benzyloxycarbonyl chloride (1.12 ml; 7.88 mmol) is added, under argon, to a solution, previously cooled to 0*C, of the acetal 7 (1.82 g; 6 mmol) obtained in step 5 in a mixture of anhydrous dichloromethane (30 ml) and 35 anhydrous triethylamine (1.27 ml; 9.1 mmol). After stirring for 8 h, the reaction medium is poured into 100 ml H 2 0 and the aqueous phase is extracted with CH 2 C1 2 (100 ml). The organic phase is dried over MgSO 4 , concen trated under reduced pressure and the residue is purified - 16 by silica gel chromatography (cyclohexane/AcOEt: 6/1 and 4/1) so as to isolate 1.9 g of 8 (72%). Characteristics: TLC: cyclohexane/AcOEt: 1/1; Rf = 0.64 - = -270 (c = 1.14; CHCl 3 ) 5 MS: m/z 438 (M + H)* m.p. = 1020C

C

2 2

H

3 5

NO

6 SiM = 437 C H N Calculated 60.38 8.06 3.20 10 Found 60.27 8.10 3.29 Step 7 4' -Benzyloxyc arbonylepipodophyllotoxin 3 -aminobenzyloxy carbonyl-2,3-dideoxy-4,6-0-ethylidene- -D-arabinohexo pyranoside (9) 15 5.06 ml of tetrabutylammonium fluoride (1.1 M solution in THF; 5.5 mmol) are added to the sugar 8 (2.0 g; 4.57 mmol) obtained in step 6 in solution in anhydrous dichloromethane (100 ml) . When the complete disappearance of 8 is observed by TLC (2 h of stirring), 20 the reaction medium is cooled to -20 0 C. DMEPT4'-OZ (2.57 g; 4.8 mmol) and then 8.44 ml of BF 3 .Et 2 O (68.6 mmol) are then added successively. After reacting for 1 h at -20 0 C, the reaction medium is poured into 200 ml of a saturated NaHCO 3 solution (addition of NaHCO 3 25 salts) (pH = 9). The organic phase is dried over MgSO 4 , concentrated under reduced pressure and then the crude residue (6.2 g) is chromatographed on a silica gel

(CH

2 Cl 2 /Acetone: 98/2 and then 97/3) to give 9 (2.1 g; 54%). 30 Characteristics: TLC: CH 2 Cl 2 /Acetone: 92/8; Rf 0.61 [ -2 _740 (c = 1.1; CHCl 3 ) MS: m/z 857 (M + NH4)* m.p. = 175 0 C

C

4 5

H

4 5

NO

1 5 M = 839 35 C H N Calculated 64.36 5.40 1.67 Found 64.21 5.30 1.58 Step 8 30 pl of triethylamine and then 150 mg of 10% - 17 palladium on carbon are added to a solution of 9 (0.28 g; 0.33 mmol) in 20 ml of ethyl acetate. The reaction medium is placed under a hydrogen atmosphere (atmospheric pressure) . After stirring for 1 h 30 min at room tempera 5 ture, the catalyst is removed by filtration and the organic phase concentrated under reduced pressure and then chromatographed on a silica gel (CH 2 Cl 2 /MeOH: 97/3 and then 95/5) to give 172 mg of the pure compound 10 (90%). (Recrystallization from CH 2 Cl 2 /pentane) . 10 Characteristics: TLC: CH 2 Cl 2 /MeOH: 95/5; Rf = 0.31 [a] 20 = -1200 (c = 1.05; CHCl 3 ) MS: m/z 594 (M + Na)+ 610 (M + K)+ m.p. = 219 0 C

C

2 9

H

3 3

NO

1 1 M = 571 15 C H N Calculated 60.94 5.82 2.45 Found 60.45 5.78 2.58 1 H NMR 300 MHz CDCl 3 5 1.36 (3H, d, J = 5Hz, CH 3 -CH); 1.51 (1H, m, H2",a); 2.05 (1H, m, H2"e); 2.88 (1H, m, H 3 ); 3.02 20 (2H, m, H 3 " and Hz.); 3.28 (1H, m, J2- 1 = 5.2 Hz, H 2 ); 3.30 (1H, m, H 5 ,); 3.57 (1H, t, J = 10 Hz, H6"a); 3.75 (6H, s,

OCH

3 ) ; 4.15 (1H, dd, J = 5 Hz, J = 10 Hz, H6"e) ; 4.41 (1H, dd, J = 9 Hz, Hia); 4.21 (1H, t, J = 9 Hz, H11b); 4.59 (1H, d, J = 5.2 Hz, H 1 ) ; 4.75 (1H, q, J = 5 Hz, H 7 .); 4.85 25 (1H, dd, J = 9 Hz, J = 2 Hz, H 1 ,,); 4.94 (1H, d, J 3.3 Hz, H 4 ); 5.98 (1H, d, OCHAO); 6.00 (1H, d, OCHBO); 6.24 (2H, s, H 2 and H 6 ,); 6.55 (1H, s, H 8 ); 6.75 (1H, s,

H

5 ). Preparation of the hydrochloride 30 A solution of 0.098 M hydrochloric methanol (1.09 ml; 0.106 mmol) is added to the amine 10 (61 mg; 0.10 mmol) in solution in anhydrous dichloromethane (6 ml). The reaction medium is stirred for 10 minutes. The expected product is precipitated after addition of 35 ether (20 ml). 56 mg (86%) of the hydrochloride of 10 are recovered. Characteristics: m.p. = 230 0 C

C

2 9

H

3 2 N0 1 1 ClM = 606 Test of solubility: 2.56 mg in 0.3 ml - 18 of water C = 0.014 M EXAMPLE 2 General formula I (NR 1

R

2 = #NMe 2 ; XY = OCH (CH 3 ) OCH 2 ) 5 4'-Demethyl-4-0(3-dimethylamino-2,3-dideoxy-4,6 ethylidene-#-D-arabinohexopyranosyl)epipodophyllotoxin (Compound 11) Formaldehyde (13.5 pl) and sodium cyanoboro hydride (85 mg) are added successively to a solution of 10 10 (0.19 g; 0.33 mmol) in anhydrous dichloromethane (15 ml). After stirring for 45 min at room temperature, these same reagents are added and the reaction is con tinued for 45 min. The reaction medium is diluted with

CH

2 Cl 2 (30 ml) and washed with water (40 ml) . The organic 15 phase is dried over MgSO 4 and concentrated under reduced pressure. The residue is chromatographed on a silica gel

(CH

2 Cl 2 /MeOH: 97/3) . This gives 101 mg of 11 (51%) . Characteristics: TLC: CH 2 Cl 2 /MeOH: 95/5; Rf = 0.4 [a] 20 = -1210 (c = 1; CHC1 3 ) 20 MS: m/z 600 (M + H)* m.p. = 270 0 C

C

31

H

37

NO

11 M = 599 1 HNMR 300 MHz CDCl3 6 1.38 (3H, d, J = 5 Hz, CH 3 -CH) ; 1.55 (1H, m, H 2 -a); 1.96 (1H, m, H 2 -a); 1.96 (1H, m, H 2 -e); 2.33 25 (3H, s, CH 3 -N); 2.91 - 2.82 (1H, m, H 3 ); 2.91 - 2.82 (1H, m, H3.) ; 3.35 - 3.25 (1H, m, H..) ; 3.38 (1H, t, J4...

5 = 9 Hz, J 4 ,-3, = 9 Hz, H 4 -) ; 3.58 (1H, t, J6-a-s- = 10 Hz, J6"a-6"e = 10 Hz, H 6 .a) ; 3.75 (3H, s, CH 3 0) ; 4.16 (1H, dd, JS-e-S- = 5 Hz, Js-e-E-a = 10 Hz, H 6 -e) ; 4.21 (1H, t, J9b-sa = 9 Hz, J 9 b- 3 30 = 8 Hz, H 9 b) 4.42 (1H, dd, J 9 a- 9 b = 9 Hz, J 9 a- 3 = 10. 5 Hz,

H

9 a) ; 4.59 (1H, d, J 1

_

2 = 5.2 Hz, H 1 ); 4.74 (1H, q, J = 5 Hz, CH-CH 3 ) ; 4.82 (1H, dd, J1-- 2 -a = 9.5 Hz, J1-- 2 -e = 2 Hz,

H

1 .); 4.95 (1H, d, J 4

_

3 = 3.2 Hz, H 4 ); 5.97 (1H, d, OCHAO); 6.00 (1H, d, OCHBO) ; 6.25 (2H, s, H 2 and H,,); 6.55 (1H, s, 35 H 8 ) ; 6.76 (1H, s, H) . Preparation of the hydrochloride A solution of 0.098 M hydrochloric methanol (1.72 ml; 0.17 mmol) is added to the amine 11 (101 mg; 0.17 mmol) in solution in anhydrous dichloromethane - 19 (7 ml) . The reaction medium is stirred for 10 minutes. The expected product is precipitated after addition of ether (20 ml). 86 mg (81%) of the hydrochloride of 11 are recovered. 5 Characteristics: m.p. = 199 0 C

C

31

H

33 N0 11 ClM = 635 Test of solubility: 2.5 mg in 0.1 ml of water C = 0.038 M 10 EXAMPLE 3 General formula I: NRjR 2 = a-NH 2 ; XY = OCH(CH 3

)OCH

2 4' -Demethyl-4-0- (3-amino-2, 3-dideoxy-4, 6-ethylidene$-D ribohexopyranosyl) epipodophyllotoxin (compound 19) Step 1 15 Tert-butyldimethylsilyl-3-azido-2, 3-dideoxy-$-D-ribohexo pyranoside (compound 12) In a manner similar to step 3 of Example 1, but using compound 4, compound 12 is obtained which is used directly in step 2. 20 Step 2 Tert-butyldimethylsilyl-3-azido-2,3-dideoxy-4,6-O ethylidene- -D-ribohexopyranoside (13) 1.1 ml (8 mmol) of acetaldehyde diethyl acetal and then 52 mg of camphorsulfonic acid (0.24 mmol) are 25 added to a solution of 12 (0.25 g; 0.8 mmol) obtained in step 1 in 10 ml of acetonitrile. The reaction medium is stirred at room temperature for 9 h and then diluted with ethyl acetate (30 ml) before washing with a solution of sodium hydrogen carbonate (pH = 9) and then with water 30 (30 ml). The organic phase is dried over MgSO 4 and then concentrated under reduced pressure to give 0.3 g of crude product. Purification on a silica gel (cyclo hexane/AcOEt: 15/1) makes it possible to isolate 0.15 g of pure 13 (55%). 35 Characteristics: TLC: cyclohexane/AcOEt: 4/1; Rf = 0.77 [a]D 0 -350 (c = 1.1 CHCl 3 )

C

14

H

27

N

3 0 4 SiM = 329 Step 3 4'-Benzyloxycarbonylepipodophyllotoxin 3-azido-2,3- - 20 dideoxy-4, 6-O-ethylidene-#-D-ribohexopyranoside (compound 14) 1st route of synthesis: Trimethylsilyl trifluoromethanesulfonate (TMSOTf) 5 (446 pl; 2.46 mmol) is added to a mixture of DMEPT 4'-OZ (438 mg; 0.82 mmol), of 13 (270 mg; 0.82 mmol) obtained in step 2 and of molecular sieve 4 A (1.5 g) in anhydrous dichloromethane (30 ml) cooled to -40 0 C. After reacting for 1 h 15 min at -400C, the reaction medium is 10 neutralized with triethylamine (342 pl), filtered and then washed with a saturated solution of NaCl (20 ml). The organic phase is dried over MgSO 4 , concentrated under reduced pressure and then the crude residue is chromato graphed on a silica gel (cyclohexane/AcOEt: 65/35) to -15 give 14 (260 mg; 45%). 2nd route of synthesis: Boron trifluoride etherate (BF 3 .Et 2 O) (425 pl; 3.46 mmol) is added to a mixture of DMEPT 4'OZ (1.85 g; 3.46 mmol), of 13 (1.20 g; 3.64 mmol) obtained in step 2 20 in anhydrous dichloromethane (100 ml) cooled to -15*C. After reacting for 2 h at -15*C, the reaction medium is diluted with 100 ml of CH 2 Cl 2 and then poured into 200 ml of a saturated solution of NaHCO 3 . The organic phase is dried over MgSO 4 , concentrated under reduced pressure and 25 then the crude residue (2.9 g) is chromatographed on silica gel (cyclohexane/AcOEt: 65/35) to give 14 (1.19 g; 47%) (recrystallization Et 2 O/hexane). Characteristics: TLC: cyclohexane/AcOEt: 6/4; Rf = 0.41 cyclohexane/AcOEt: 65/3; Rf = 0.27 30 [a] = -105* (c = 1.05; CHCl 3 ) MS: m/z 749 (M + NH 4 )* m.p. = 1.390C

C

3 7

H

3 7

N

3 0, 3 M = 731 Step 4 35 Epipodophyllotixin 3 -amino-2, 3 -dideoxy-4, 6 -0-ethylidene P-D-ribohexopyranoside 15 20 pl of triethylamine and then 20 mg of 10% palladium on carbon are added to a solution of 14 (110 mg; 0.15 mmol) obtained in step 3 in a mixture of - 21 10 ml of ethanol and 5 ml of ethyl acetate. The reaction medium is placed under a hydrogen atmosphere (atmospheric pressure) . After stirring for 2 h at room temperature, the catalyst is removed by filtration and the organic 5 phase concentrated under reduced pressure and then chromatographed on a silica gel (CH 2 Cl 2 /MeOH: 97/3 and then 95/5) to give 63 mg of pure 15 (72%). Characteristics: TLC: CH 2 Cl 2 /MeOH: 95/5; Rf = 0.39 [a] 20 = -100o (c = 1.05; CHCl 3 ) 10 MS: m/z 572 (M + 1) 589 (M + NHg)+ m.p. = 2170C

C

2 9

H

3 3 N0 1 1 M = 571 1 HNMR 300 MHz CDCl 3 8: 1.35 (3H, d, J = 5 Hz, CH 3 -CH) ; 1.73 (1H, m, H2- a) 1-90 (1H, m, H2 "e); 2.83 (1H, m, H3); 15 3.22 (1H, dd, J 2

-

1 = 5.2 Hz, J 2

-

3 = 14 Hz, H 2 ); 3.42 (1H, dd, J 4

._

3 , = 9.5 Hz, J 4 -- 5,, = 3.5 Hz, H 4 ,,); 3.74 (6H, s,

OCH

3 ); 3.49 - 3.60 (2H, m, H 3 ,, and H 6 ,,); 3.94 - 4.02 (1H, m, H 5 ,,) ; 4.10 - 4.20 (2H, m, H 6 ,, and H11b) ; 4.42 (1H, dd, J1la-3 = 9 Hz, J11a-11b = 9.5 Hz, H11a) ; 4.57 (1H, d, J 1 -2 = 20 5.2 Hz, Hj); 4.78 (1H, q, J = 5 Hz, H 7 ,,); 4.91 (1H, d, J4-3 = 3.4 Hz, H 4 ); 5.38 (1H, dd, Jl'- 2 "e = 2 Hz, J1"- 2 "a = 9 Hz, H 1 ,.); 5.93 (1H, s, OCHAO); 5.97 (1H, s, OCHBO); 6.25 (2H, s, H 2 , and H 6 ,); 6.51 (lH, s, H 8 ); 6.86 (1H, s, H 5 ). Preparation of the hydrochloride 25 890 l of a 0.098 M hydrochloric methanol sol ution (0.087 mmol) are added to the amine 15 (50 mg; 0.087 mmol) obtained in step 4. After stirring for 10 minutes and addition of ether (10 ml), the crystals or hydrochloride (52 mg, 98%) are obtained by filtration. 30 Characteristics: m.p. = 175 0 C

C

2 9

H

3 4

NO

1 ClM = 607 Test of solubility: 2.2 mg in 0.2 ml of water C = 0.02 M 35 EXAMPLE 4 General formula I: NR 1

R

2 = a-NMe 2 ; XY = OCH(CH 3

)OCH

2 4'-Demethyl-4-0(3-N,N-dimethylamino-2,3-dideoxy-4,6-0 ethylidene--D-ribohexopyranosyl) epipodophyllotoxin 20 Formaldehyde (10.3 l) and sodium cyanoboro- - 22 hydride (12 mg) are added successively to a solution of 15 (29 mg: 0.05 mmol) obtained in step 4 of Example 3 in 1 ml of acetonitrile. After stirring for 2 h at room temperature, the reaction medium is diluted with CH 2 Cl 2 5 (20 ml) and washed with water (20 ml). The organic phase is dried over MgSO 4 and concentrated under reduced pressure. The residue is again added to the reaction under the same conditions and after an identical treat ment chromatographed on a silica gel (CH 2 Cl 2 /MeOH: 97/3). 10 This gives 29 mg of 20 (95%). Characteristics: TLC: CH 2 Cl 2 /MeOH: 95/5; Rf = 0.5 [ -850 (c = 1.06; CHCl 3 ) MS: m/z 600 (M + H)+ m.p. = 1400C 15 C 3 1

H

3 7

NO

1 1 M = 599 1H NMR 300 MHz CDCl 3 8: 1.35 (3H, d, J = 5 Hz); 1.55 (lH, m, H2"a); 2.20 (lH, m, H2"e); 2.36 (3H, s, CH 3 N) ; 2.62 (lH, m, 2.62, H 3 ) ; 2.85 (lH, m, H 3 ) ; 3.23 (lH, dd, J 2 -1 = 5.2 Hz, J 2

-

3 = 14 Hz, H 2 ) ; 3.52 (lH, dd, J 4

"-

3 = 3 Hz, 20 J 4

-

3 ." = 9 Hz, H 4 ,) ; 3.61 (lH, t, J6"a-6"e 10 Hz, J6"a-5" = 10 Hz, H6-a); 3.75 (3H, s, CH 3 0); 4.22 - 4.02 (lH, m,

H

5 ") ; 4.22 - 4.02 (lH, m, H6"-e); 4.22 - 4.02 (1H, m, H 9 b); 4.43 (lH, dd, J9a-9b = 9 Hz, J 9 b- 3 = 10.5 Hz, H 9 b); 4.62 4.57 (lH, m, J 1

-

2 = 5.2 Hz, Hj); 4.62 - 4.57 (3H, m, J = 25 5 Hz, CH-CH3), 4.88 (lH, d, J 4

-

3 = 3.4 Hz, H 4 ); 5.96 (1H, d, OCHAO); 5.98 (lH, d, OCHBO); 6.25 (2H, s, H 2 and H 6 '); 6.52 (1H, s, H 8 ); 6.80 (lH, s, H 5

)

Preparation of the hydrochloride A 0.098 M hydrochloric methanol solution is added 30 to the amine 20 (59 mg; 0.1 mmol) in solution in anhydrous methanol (2 ml) (1 ml; 0.1 mmol). The reaction medium is stirred for 10 minutes. The expected product is precipitated after addition of ether (20 ml). 33 mg (53%) of crystals are recovered. 35 Characteristics: m.p. = 1500C

C

3 1

H

3 8

NO

1 1 ClM = 635 Test of solubility: 2.6 mg in 0.1 ml of water C = 0.04 M - 23 EXAMPLE 5 General formula I: NR 1

R

2 = a-NHCH 2 CN; XY = OCH(CH 3

)OCH

2 4'-Demethyl-4-0-(3-cyanomethylamino-2,3-dideoxy-4,6 ethylidene-$-D-ribohexopyranosyl)epipodophyllotoxin 5 (Compound 16) 200 pl of triethylamine (1.47 mmol) and then 100 gl of iodoacetonitrile (1.47 mmol) are added to a solution of 15 (120 mg; 0.21 mmol) obtained in step 4 of Example 3 in 4 ml of dimethylformamide. The reaction 10 medium is stirred for 20 h at room temperature and then diluted with ethyl acetate (30 ml) before washing with water (4 x 30 ml). The organic phase is dried over MgSO 4 , concentrated under reduced pressure and then chromato graphed on a silica gel (CH 2 Cl 2 /acetone: 92/8) to give 15 71 mg of pure 16 (55%). Characteristics: TLC: CH 2 Cl 2 /MeOH: 95/5; Rf = 0.25 [a]2 0 = -86O (c = 0.80; CHCl 3 ) MS: m/z 611 (M + H)+ 628 (M + NH4)*

C

3 1

H

34

N

2 0 11 M = 610 20 1 H NMR 300 MHz CDCl 3 8: 1.35 (3H, d, J = 5Hz, CH 3 CH) ; 1.69 (1H, m, J2"a-1 = 9.5 Hz, J2"a-2"e = 13 Hz, J 2 "a- 3 = 3 Hz, H2"a); 1.93 (1H, m, H2"e); 2.86 (lH, m, H 3 ); 3.25 (1H, dd, J2-1 = 5.2 Hz, J 2

-

3 = 14 Hz, H 2 ); 3.51 - 3.56 (2H, m, H 4 . and H6-a); 3.56 (2H, m, CH 2 CN); 3.76 (6H, s, OCH 3 ); 3.88 25 (1H, m, H 5 ") ; 4.17 (2H, t, J1la- 3 = 8 Hz, J11a-11b = 10 Hz,

H

3 and H11b); 4.14 (1H, dd, J6a"-6"b = 10 Hz, J6"b-5" = 5 Hz, H6"b) ; 4.41 (1H, dd, J1la- 3 = 9 Hz, J11a-11b = 10 Hz, H1ia); 4.60 (1H, d, J 1

-

2 = 5.3 Hz, H); 4.75 (1H, q, J = 5 Hz, H 7 ,,); 4.90 (1H, d, J 4

-

3 = 3.3 Hz, H 4 ); 5.17 (1H, dd, 30 Ji"-2"c = 2 Hz, Jl"-2"a = 9.5 Hz, H 1 .); 5.99 (1H, s, OCHAO); 6.00 (1H, s, OCHBO); 6.25 (2H, s, H2 and H 6 ,); 6.54 (1H, s, H 8 ); 6.79 (1H, s, H 5 ). EXAMPLE 6 General formula I: NR 1

R

2 = a-morpholino; XY = OCH(CH 3

)OCH

2 35 4'-Demethyl-4-0-(3-N-morpholino-2,3-dideoxy-4,6 ethylidene-$-D-ribohexopyranosyl)epipodophyllotoxin (Compound 17) 58 pl of triethylamine (0.42 mmol) and then 512 mg of diiodoethyl ether (1.57 mmol) are added to a - 24 solution of 15 (60 mg; 0.10 mmol) obtained in step 4 of Example 3 in 2 ml of dimethylformamide. The reaction medium is stirred for 96 h at room temperature and in the dark and then diluted with ethyl acetate (30 ml) before 5 washing with water (4 x 30 ml). The organic phase is dried over MgSO 4 , concentrated under reduced pressure and then chromatographed on a silica gel (CH 2 Cl 2 /acetone: 92/8) to give 46 mg of pure 17 (68%). Characteristics: TLC: CH 2 Cl 2 /acetone: 92/8; Rf = 0.31 10 [a]2 = -980 (c = 1.04; CHCl 3 )

C

3 3

H

3 9

NO

1 2 M = 641 H NMR 300 MHz CDC1 3 8: 1.33 (3H, d, J = 5Hz, CH 3 CH) ; 1.55 (1H, m, J2'a-1. = 9.5 Hz, J2"a-2"e = 13 Hz, J 2 "a- 3 " = 3 Hz, H2"a); 2.15 (1H, m, H2-e); 2.84-2.90 (5H, m, CH 2 N and H 3 ); 15 2.80 (1H, dd, J 3

--

4 ., = 3Hz, J3"- 2 "a = 3 Hz, J3"- 2 "e = 3 Hz,

H

3 -. ); 3.23 (lH, dd, J 2

-

1 = 5.2 Hz, J 2

-

3 = 14 Hz, H 2 ); 3.48 (1H, t, J6"a-6"b = 12 Hz, J 6 "a-5" = 12 Hz, H6"a) ; 3.57 (lH, dd, J 3

"-

4 . = 3Hz, J 4 "-5. = 9 Hz, H 4 .,); 3.66-3.76 (10H, m,

OCH

3 and OCH 2 ); 4.08-4.16 (2H, m, H 5 . and H6"b); 4.20 (1H, 20 t, J1la- 3 = 8 Hz, J11a-11b = 9 Hz, Hia) ; 4.43 (lH, dd, J11b- 3 = 9 Hz, J11b-11a = 9 Hz, H11b) ; 4.57-4.62 (2H, m, H, and H 7 .,) ; 4.89 (1H, d, J 4

-

3 = 3.4 Hz, H 4 ) ; 5.20 (lH, dd, J"-2"le = 2 Hz, J1- 2 'a = 9.5 Hz, H 1 .) ; 5.97 (1H, s, OCHAO) ; 6.00 (1H, s, OCHBO); 6.25 (2H, s, H 2 , and H 6 ,); 6.54 (1H, 25 s, H 8 ); 6.73 (1H, s, H 5 ) EXAMPLE 7 General formula I: NR 1

R

2 = a-NH 2

(CH

2 ) 2

NH

2 ; XY OCH (CH 3 ) OCH 2 4'-Demethyl-4-O [3- (2-Aminoethylamino) -2, 3-dideoxy-4, 6 30 ethylidene-$-D-ribohexopyranosyl] epipodophyllotoxin 18 Triethylamine (127 pl; 0.91 mmol) and N-benzyl oxycarbonyl-2-iodoethylamine (0.28 g; 0.91 mmol) are added to a solution of 15 (173 mg; 0.30 mmol) obtained in step 4 of Example 3 in 10 ml of dimethylformamide. The 35 reaction medium is stirred for 5 days at room temperature and then diluted with water (30 ml). After extraction with ethyl acetate (30 ml), washing with water (5 x 20 ml), the organic phase is dried over MgSO 4 , concentrated under reduced pressure and then chromato- - 25 graphed on a silica gel (CH 2 Cl 2 /MeOH: 97/3) to give 155 mg of pure 18 (68%). Characteristics: TLC: CH 2 Cl 2 /MeOH: 95/5; Rf = 0.70 [a]D 0 -740 (c = 1.17; CHCl 3 ) 5 MS: m/z 749 (M + H)+

C

3 9

H

4 4

N

2 0 1 3 M = 748 which is used directly in the following debenzylation step: Triethylamine (30 pl) and then 10% palladium on 10 carbon (0.1 g) are added to a solution of 18 (0.15 g; 0.20 mmol) in a mixture of ethyl acetate and ethanol (10 ml, 1/1). The reaction medium is placed under a hydrogen atmosphere (atmospheric pressure). After stirring for 1 h 30 min at room temperature 15 in the presence of hydrogen at atmospheric pressure, the catalyst is removed by filtration and the organic phase concentrated under reduced pressure and chromatographed on a silica gel (CH 2 Cl 2 /MeOH(NH 3 ): 97/3) to give 107 mg (84%) of 19. 20 Characteristics: TLC: CH 2 Cl 2 /MeOH(NH 3 ) : 95/5; Rf = 0.22 - -770 (c = 1; CHC1 3 ) m.p. = 130 0 C

C

3 1

H

3 8 N0 1 1 M = 614 1 H NMR 300 MHz CDCl 3 8: 1.33 (3H, d, J = 5 Hz); 1.55 (1H, 25 m, H2"a) ; 2.15 (1H, m, H2"e) ; 2.33 (3H, m, NH 2 and NH, exchangeable); 2.84-2.90 (5H, m, CH 2 -N and H 3 ); 3.20 (1H, dd, J 3

--

4 ' = 3 Hz, J 3

-

2 -a = 3 Hz, J3"-2"e = 3 Hz, H 3 .) ; 3 .23 (1H, dd, H 2 ); 3.48 (1H, t, J6"a-6"b = 12 Hz, J 6 "a-5" 12 Hz, H6"a) ; 3.57 (1H, dd, J 4 -- 3 = 3 Hz, J 4 "-5,, = 9 Hz, 30 Hg.,); 3.75 (6H, s, OCH 3 ); 4.08-4.16 (2H, m, H 5 . and H 6 "b); 4.20 (1H, t, J11b- 3 = 8, J11b-11a = 9 Hz, Hilb) ; 4.43 (1H, dd, J1la- 3 = 9 Hz, J11a-11b = 9 Hz, Hila); 4.57-4.62 (2H, m, H, and H 7 .); 4.89 (1H, d, J 4

-

3 = 3.4 Hz, H 4 ); 5.20 (1H, dd, Jl"-2"e = 2Hz, J1"-2"a = 9.5 Hz, H 1 .); 5.97 and 6.00 35 (2H, d, OCH 2 0); 6.25 (2H, s, H 2 , and H 6 ,); 6.54 (1H, s,

H

8 ); 6.73 (1H, s, H 5 ). Preparation of the hydrochloride A 0.098 M hydrochloric methanol solution (2.13 ml; 0.21 mmol) is added to the diamine 19 (64 mg; - 26 0.10 mmol) in solution in anhydrous methanol (3 ml). The reaction medium is stirred for 10 minutes. The expected product is precipitated after addition of ether (20 ml). 50 mg (73%) of the hydrochloride are recovered. 5 Characteristics: m.p. = 170 0 C

C

3 1

H

3 9

N

2 0 1 1 Cl 2 M = 649 Test of solubility: 2.0 mg in 0.05 ml of water C = 0.06 M 10 EXAMPLE 8 General formula I: NRjR 2 = a-NH 2 ; X = OH; Y = CH 3 4' -Demethyl-4-0 (3-amino-2, 3, 6-trideoxy -D-ribohexo pyranosyl) epipodophyllotoxin (compound 26) Step 1 15 Tert-butyldimethylsilyl-3-azido-2, 3-dideoxy-6-0-tosyl-$ D-ribohexopyranoside 21 A solution of tosyl chloride (1.55 g; 8.15 mmol) in pyridine (10 ml) is added dropwise to a solution of the diol 12 (2.06 g; 6.79 mmol) obtained in step 1 of 20 Example 3 previously cooled to 0 0 C. After stirring at the same temperature for 1 h and then for 18 h at 20 0 C, the reaction medium is diluted with dichloromethane (100 ml). The organic phase is washed with water (2 x 100 ml), dried over MgSO 4 and concentrated under reduced pressure. 25 The residue is purified on a silica gel (cyclohexane/ EtOAc: 8/2). This gives 2.02 g of 21 (65%). Characteristics: TLC: cyclohexane/AcOEt: 2/1; Rf = 0.46 MS: m/z 475 (M + H)+

C

1 9

H

3 1

N

3 0 6 SSiM = 457 30 Step 2 Tert-butyldimethylsilyl-3-azido-6-iodo-2, 3, 6-trideoxy-$ D-ribohexopyranoside 22 The compound 21 (2.02 g; 4.42 mmol) obtained in step 1 in solution in 120 ml of acetone is heated under 35 ref lux for 72 h in the presence of sodium iodide (2.65 g; 17.68 mmol) . After cooling, the reaction medium is concentrated under reduced pressure (30 ml) and then diluted with dichloromethane (100 ml). After washing with a 10% aqueous solution of sodium thiosulfate and then - 27 drying over MgSO 4 and evaporating under reduced pressure, a residue is obtained which is purified on silica (cyclo hexane/EtOAc: 8/2) giving 1.5 g (82%) of 22. Characteristics: [X]20 = -300 (c = 1.06, CHCl 3 ) 5 MS: m/z 431 (M + NH 4 )+

C

1 2

H

2 4

N

3 0 3 ISiM = 413 C H N Calculated 34.87 5.81 10.17 Found 35.07 5.76 10.25 10 Step 3 Tert-butyldimethylsilyl-3-azido-6-iodo-4-0-chloroacetyl 2,3, 6-trideoxy-$-D-ribohexopyranoside (compound 23) Chloroacetyl chloride (396 ml; 5 mmol) is added to a solution of 22 (1.03 g; 2.5 mmol) obtained in the 15 preceding step in a mixture of dichloromethane (20 ml) and pyridine (404 pl; 5 mmol). After stirring for 1 h at -10 0 C, the reaction medium is diluted with CH 2 C1 2 (30 ml) and washed with water (3 x 20 ml). A customary treatment, followed by chromatography on silica (cyclohexane/EtOAc: 20 10/1) gives 1.1 g (90%) of compound 23. Characteristics: [a]20 _14o (c = 1.03, CHCl 3 ) MS: m/z 507 (M + NH4)

C

1 4

H

2 5

N

3 0 4 ClISiM = 489 C H N 25 Calculated 34.35 5.11 8.59 Found 34.69 5.16 8.22 Step 4 4'-Demethyl-4-O(3-azido-6-iodo-2,3,6-trideoxy-$-D-ribo hexopyranosyl) epipodophyllotoxin (compound 24) 30 Boron trifluoride etherate (BF 3 .Et 2 O) (455 l; 3.7 mmol) is added to a mixture of DMEPT 4'-OZ (1 g; 1.85 mmol), of 23 (1 g; 2.04 mmol) from the preceding step in anhydrous dichloromethane (100 ml) cooled to -15 0 C. After reacting for 5 h (-15 0 C -> OOC), the reaction 35 medium is diluted with CH 2 Cl 2 (100 ml) and then poured into a saturated NaHCO 3 solution (200 ml). The organic phase is dried over MgSO 4 , concentrated under reduced pressure. The crude residue is chromatographed on a silica gel (cyclohexane/AcOEt: 7/3) to give 24 (0.8 g; - 28 48%) Characteristics: TLC: cyclohexane/AcOEt: 1/1; Rf = 0.58 [a]D 0 -850 (c = 1.26; CHCl 3 ) MS: m/z 909 (M + NH 4 )* 5 m.p. = 1430C

C

3 7

H

3 5

N

3 0 1 3 ClIM = 891 Step 5 4'-Demethyl-4-O(3-Azido-6-iodo-2,3,6-trideoxy-$-D-ribo hexopyranosyl)4'-benzyloxycarbonylepipodophyllotoxin 10 (compound 25) OH~ resin (Amberlite IRA 410) is added to a solution of the azidoglycoside 24 (257 mg; 0.29 mmol) in a CH 2 Cl 2 /MeOH mixture (15 ml, 2/1). After reacting for 3 h at 20 0 C, the reaction medium is filtered and then 15 concentrated under reduced pressure to give 0.22 g of pure 25 (94%). Characteristics: TLC: cyclohexane/AcOEt: 1/1; Rf = 0.46 [a]2 0 = -570 (c = 1.02; CHCl 3 ) MS: m/z 833 (M + NH4)+ 20 m.p. = 115 0 C

C

3 5

H

3 4

N

3 0 12 IM = 815 Step 6 4'-Demethyl-4-0(3-amino-2,3,6-trideoxy-$-D-ribohexo pyranosyl) epipodophyllotoxin (compound 26) 25 Triethylamine (100 pl) and then 10% palladium on carbon (0.2 g) are added to a solution of 25 (0.20 g; 0.25 mmol) from the preceding step in 10 ml of ethyl acetate. After stirring for 30 h at room temperature in the presence of hydrogen at atmospheric pressure, the 30 catalyst is removed by filtration and the organic phase concentrated under reduced pressure and then chromato graphed on a silica gel (CH 2 Cl 2 /MeOH: 92/8) to give 50 mg (38%) of 26. Characteristics: TLC: CH 2 Cl 2 /MeOH: 90/10; Rf = 0.33 35 [a]2 0 = -9o (c = 0.5; CHCl 3 ) MS: m/z 530 (M + H)+ m.p. = 140 0 C

C

2 7

H

3 1 NOgM = 529 - 29 Preparation of the hydrochloride A 0.098 M hydrochloric methanol solution (1.44 ml; 0.14 mmol) is added to the preceding amine 26 (70 mg; 0.14 mmol) in solution in anhydrous methanol 5 (6 ml). The reaction medium is stirred for 10 minutes. The expected product is precipitated after addition of ether (20 ml). 40 mg (53%) of the crystallized hydrochloride are recovered. Characteristics: m.p. = 1640C 10 C 2 7

H

3 2

NO

1 0ClM = 565 Test of solubility: 2.6 mg in 0.2 ml of water C = 0.02 M EXAMPLE 9 15 General formula I: NR 1

R

2 = a-NH 2 ; X = OH; Y = CH 2

NH

2 4' -Demethyl-4-0- (3, 6-diamino-2, 3, 6-trideoxy-$-D-ribohexo pyranosyl) epipodophyllotixin (compound 29) Step 1 4'-Demethyl-4'-benzyloxycarbonyl-4-0(3,6-diazido-4 20 azidoacetyl-2, 3, 6-trideoxy- -D-ribohexopyranosyl) epipodo phyllotoxin (compound 27) Sodium azide (0.1 g; 1.5 mmol) is added to a solution of 24 (0.45 g; 0.51 mmol) obtained in step 4 of Example 8 in 10 ml of dimethylformamide. The reaction 25 medium is stirred for 64 h at room temperature, diluted with water (30 ml) and ethyl acetate (30 ml) . The organic phase is washed with water (4 x 20 ml), dried over MgSO 4 , concentrated under reduced pressure and chromatographed on silica gel (cyclohexane/AcOEt: 7/3) to give 0.36 g of 30 27 (90%). Characteristics: TLC: cyclohexane/AcOEt: 6/4; Rf = 0.44 [ = -640 (c = 1; CHCl 3 ) MS: m/z 831 (M + NH 4 )* m.p. = 1200C 35 C 3 7

H

3 5

N

9 0 1 3 M = 813 Step 2 4' -Demethyl-4' -benzyloxycarbonyl-4-0 (3, 6-diazido-2, 3,6 trideoxy- -D-ribohexopyranosyl)epipodophyllotoxin (compound 28) - 30 OH~ resin (Amberlite IRA 410) is added to a solution of the azidoglycoside 27 (70 mg; 0.08 mmol) in 3 ml of a CH2Cl2/MeOH mixture (2/1, v/v). After reacting for 5 h at 200C, the reaction medium is filtered and then 5 concentrated under reduced pressure to give 59 mg of pure 28 (94%). Characteristics: TLC: cyclohexane/AcOEt: 6/4; Rf = 0.25 [a] 20 -530 (c = 1.04; CHCl 3 ) m.p. = 1250C 10 C 3 5

H

3 4

N

6 0 1 2 IM = 730 Step 3 4' -Demethyl-4-0 (3, 6-diamino-2, 3, 6-trideoxy- -D-ribohexo pyranosyl) epipodophyllotoxin (compound 29) Triethylamine (20 gl) and then 10% palladium on 15 carbon (70 mg) are added to a solution of 28 (0.11 g; 0.15 mmol) from the preceding step, in a mixture of ethyl acetate and ethanol (10 ml, 1/1). After stirring for 16 h at room temperature in the presence of hydrogen at atmospheric pressure, the catalyst is removed by fil 20 tration and the organic phase concentrated under reduced pressure to give 78 mg (95%) of 29. Characteristics: TLC: CH 2 Cl 2 /MeOH(NH 3 ) : 90/10; Rf = 0.06

C

2 7

H

3 2

N

2 0 1 0 M = 544 25 Preparation of the dihydrochloride A 0.098 M hydrochloric methanol solution (2.92 ml; 0.28 mmol) is added to the preceding diamine 29 (78 mg; 0.14 mmol) in solution in anhydrous methanol (2 ml). The reaction medium is stirred for 10 minutes. 30 The expected product is precipitated after addition of ether (20 ml). 70 mg (79%) of crystallized dihydro chloride are recovered. . Characteristics: m.p. = 95 0 C

C

2 7

H

3 4

N

2 0 1 0 C1 2 M = 616 35 Test of solubility: 2.1 mg in 0.05 ml of water C = 0.07 M Biological experiment The molecules were tested using a biological - 31 experiment and showed their usefulness as anticancer agent in tests for leukemia P 388 in vivo in mice. This test is commonly used in the field of research on anticancer substances (Protocols for screening chemical 5 agents and natural products against animal tumors and other biological systems, R. Geran, N.H. Greenberg, M.M. MacDonald, A.M. Schumacher and B.J. Abbott, Cancer Chemotherapy reports 1972, 3, No. 2). However, this experimental model is extremely 10 chemosensitive and a very large number of compounds manifest good activity, which makes this test poorly discriminatory. We modified the test protocol to make it more selective. The tumor cells are administered by the 15 intravenous route and not by the intraperitoneal route. They are thus rapidly distributed by the circulation in the whole body. The test product is then administered by the intraperitoneal route. Two parameters are defined in order to demonstrate the activity of the compounds: 20 - determination of the median effective dose (ED 50 ) which represents the minimum single dose of the compound to be administered in order to obtain a significant survival of the animals compared with the untreated control animals; 25 - determination of the maximum survival time of the animals regardless of the dose administered by a single injection. Being able to administer a large dose of the compound and to observe a high survival makes it possible to obtain a measurement of the 30 maximum therapeutic efficacy of the product which can be achieved. Origin of the tumor The leukemia P 388 was chemically induced in 1955 with 3-methylcholanthrene in a DBA/2 mouse (Am. J. 35 Pathol. 33, 603, 1957). Pharmacological procedure The tumors are maintained by weekly passages in asecitic form in the peritonum of a DBA/2 mouse (original line) and the experiments are carried out on CDF, hybrid - 32 female mice (bal b/c females XDBA/2 males) of 20 ± 2 g (Cancer chemother. Rep. 3, 9, 1972). The tumor cells are implanted by the intravenous route (106 cells per mouse) on day 0. The animals are randomized and distributed in 5 groups of 2 for each series. The antitumor substances are administered intra peritoneally (ip) one day after the inoculation of the leukemic cells (acute treatment). The solutions are injected in an amount of 10 ml/kg of mouse. The criterion 10 for evaluation of the antitumor activity is the extension of the survival of the treated animals. 86% of the mice die on the 7th day after the tumor transplant. A sub stance will be considered to be active if it induces a survival greater than 8 days. 15 The following table makes it possible to show the aqueous solubility of the products of the invention, expressed in mg/ml, the activity of these compounds in terms of ED 5 0 , the survival expressed in days or in T/C %, which represents the ratio of the mean survival of 20 the group of treated animals to the mean survival of the group of control animals. Aqueous P 388 Maximum T/C % solubility ED 50 (mg/kg) survival (D) mg/ml Control animals with 6-8 iv administration of (median 7) the tumor cells 25 Compounds Etoposide 0.01 10 19 271 Teniposide < 0.01 20 15 214 15 13 10 21 300 11 25 10 17 242 30 10 7 10 14 200 20 25 20 18 257 It thus appears that the compounds of the inven tion retained the level of activity of the reference compounds such as Etoposide and have, in addition, the - 33 advantage of having an advantageous aqueous solubility for formulation and administration.

Claims (11)

1. A compound of general formula I Y R- 0 04 OO OH in which the group in 3" N(Rj R 2 ) is in the $-position (2-deoxy D Arabino series) or a-position (2-deoxy D ribo 5 series) in relation to the ring, R 1 and R 2 which are identical or different, represent a hydrogen atom, a C 1 to C 6 alkyl group, capable of forming a ring, it being possible for this ring to contain a heteroatom such as oxygen or nitrogen, a C 1 to C 6 aminoalkyl group or a 10 cyanomethyl group, X and Y may be identical or different and represent OH, CH 3 , CH 2 -NH 2 , X and Y may also be linked and may con stitute a ring, such as for example a 2-methyl-1,3 dioxane, thus forming a bicyclic glycoside backbone of 15 the 4, 6-ethylidene-3 -amino-2, 3 -dideoxyglycos ide type, and its addition salts with pharmaceutically acceptable inorganic or organic acids.

2. A compound of general formula I according to Claim 1, characterized in that the group NR 1 R 2 is an NH 2 20 or N(CH 3 ) 2 group.

3. A compound of general formula I according to Claim 1, characterized in that the group NR 1 R 2 represents an amino group substituted once or twice with a methyl, CH 2 CN or a CH 2 -CH 2 -NH 2 . 25

4. A compound of the general formula I of Claims 1 to 3, characterized in that X and Y form a ring with a - 35 linkage OCH(CH 3 )OCH2

5. A compound of general formula I according to one of Claims 1 to 4, characterized in that it is chosen from the following compounds: 5 - 4'-demethyl-4-0 (3-amino-4,6-ethylidene-2,3-dideoxy $-D-arabinohexopyranosyl) epipodophyllotoxin, - 4'-demethyl-4-0 (3-amino-4, 6-ethylidene-2,3-dideoxy P-D-ribohexopyranosyl)epipodophyllotoxin, - 4 ' -demethyl-4-0 (3-dimethylamino-4, 6-ethylidene-2, 3 10 dideoxy- -D-arabinohexopyranosyl) epipodophyllotoxin, - 4'-demethyl-4-0 (3-dimethylamino-4,6-ethylidene-2,3 dideoxy- -D-ribohexopyranosyl) epipodophyllotoxin, - 4'-demethyl-4-0 (3-cyanomethylamino-4, 6-ethylidene 2,3-dideoxy-$-D-ribohexopyranosyl)epipodophyllo 15 toxin, - 4'-demethyl-4-0(3-(N-morpholino) -4,6-ethylidene-2,3 dideoxy-$-D-ribohexopyranosyl) epipodophyllotoxin, - 4' -demethyl-4-0 (3 (2-aminoethylamino) -4, 6-ethylidene 2,3-dideoxy- -D-ribohexopyranosyl)]epipodophyllo 20 toxin, - 4' -demethyl-4-0 (3-amino-2, 3, 6-trideoxy-S-D-ribohexo pyranosyl) epipodophyllotoxin, - 4'-demethyl-4-0(3,6-diamino-2,3,6-trideoxy-$-D ribohexopyranosyl) epipodophyllotoxin. 25

6. Compounds according to Claim 4, characterized in that they are provided in the form of hydrochlorides.

7. Method of preparing a compound of general formula I according to Claims 1 to 4, characterized in that a compound of formula III or IV or V IV V 30 is reacted with 4'-Demethyl-4'-benzyloxycarbonylepipodo phyllotoxin with BF 3 etherate, or trimethylsilyl tri fluoromethanesulforate in an inert solvent at low temperature; - 36 in formula III and IV, the substituent at the 3-position may be a or $, NR 1 R 2 may be an amino protected with a group Z, in formula V, P represents an alcohol-protecting 5 group and the resulting products of this condensation are deprotected and hydrogenated to give the compounds of formula I, the primary amines in the 3-position of the glycosyl are methylated with formalin and sodium cyanoborohydride. 10

8. Method of preparing a compound of formula IV, characterized in that a mixture of diacetoxyazido glycoside VI AcO MOH 11 3 V1 with tert-butyldimethylsilyl chloride in the presence of imidazole, in that the products resulting from this 15 reaction are separated, in that each of these products are deacetylated, cyclized to 4,6-ethylidene with the acetal of acetaldehyde in a catalytic acid medium.

9. As a drug, the compounds of formula I according to Claims 1 to 6. 20

10. The pharmaceutical compositions characterized in that they contain at least one compound of formula I according to one of Claims 1 to 6 and an appropriate excipient.

11. Use of a compound of formula I, according to one 25 of Claims 1 to 4, for the preparation of a drug for anticancer treatment and, in particular, for treating the forms of cancer such as, for example, small cell lung cancer, cancer of the testicles, embryonic tumors, neuroblastomas, cancer of the kidney, placental chorio 30 carcinomas, mammary adenocarcinomas, colorectal cancers, melanomas, Hodgkin's and non-Hodgkin's lymphomas, and acute leukemias.