CA2234484A1 - Novel amine derivatives of epipodophyllotoxin 2", 3"-dideoxyglycosides, preparation method therefor and use thereof as a drug and for treating cancer - Google Patents

Abstract

A compound of general formula (I), wherein X, Y, R¿1? and R¿2? are as defined in the description, a preparation method therefor and the use thereof as a drug and for treating cancer, are disclosed.

Description

CA 02234484 l998-04-08 WO 97/13776 PCT ~ R96 / 01588 New amino derivatives of 2 ", 3" dideoxyglycosides of epipodophyllotoxin, their process of prep ~ r ~ tion, their use ~ tion as medicine and their use for cancer treatments.
The present invention relates to new 2 ", 3" Arnino derivatives.
S epideodophyllotoxin dideoxyglycosides, their preparation process, their use comrne mél1ic ~ ment and their use intended for anticancer treatment.
Among the derivatives derived from natural l ~ n ~ s, there is the class of epipodophylloids, having the basic skeleton of podophyllotoxin. There are the hemisynthetic derivatives such as Etoposide or Teniposide which are used from 10 usual way in the preparation of mé ~ mentc for the tr ~ it ~ m ~? Nt of cancer. They are considered to be major products in this field.
Etoposide has ~ nfitllm ~ properties and perrnet to treat especially small cell lung cancer and testicular cancer.
The downside of these products is their lack of water solubility and meet 15 consequently riffic - ltes in terms of formulation and minictration.
The object of the present invention is to show that derivatives of 4'-demethyl epipodophyllotoxin, having in position 4, a substitution of structure 2 "-deoxyglycoside, allows, by the incorporation of one or more nitrogen, to form compounds whose addition salts have an aqueous solubility which makes it possible to respond to the problem and show the desired anticancer activity.
Patent EP-0 196 6 l 8 relates to water-soluble derivatives of 4'-demethyl epipodophyllotoxin of formula:

O ~
<~~ o .. ~ o ~ MeO ~ OM-OEI
25 where R1 = Me, Xl = NH2, NMe2, X2 = OH

WO 97/13776 PCT ~ FR96 / 01588 Patent EP-0 415 453 mentions derivatives, BD-altrosides of 4'-demethyl epipodophyllotoxin of formula I ~ O ~ 2 0 ~ 0 ~ O
<~~ 0 , ~
he MeO ~ O Me OH

where R1 = Me R2, R3 = OH and NH2 or F and NH2 as well as JP 0161, 423.
Other publications mention analogous derivatives (Carbohydr. Res. 1990, 206, 219; Chem. Pharm. Bull. 1986, 34, 3733; Chem. Pharm. Bull 1986, 34, 3741;
Chem. Lett. 1987, 799).
The fact of using a glycoside having a 2 "-deoxy position is completely particular. It makes it possible to obtain more lipophilic compounds than the analogues hydroxylated and, therefore, have a broad spectrum of activity ~ ntitnmoral. That them allows better membrane penetration and can reach more easily the biological target like, for example, solid tumors with little irrigation Furthermore, the advantage of having an amino function, for example in the 3 "position, confers a possibility of salification therefore of water solubility s ~ ffic ~ nte for better formulation and better ~ lminictration.

WO 97/13776 PCT ~ R96 / 01588 The p ~ esel, the invention therefore relates to a compound of general formula I

~, N
<O ~ o MeO ~ O Me OH

5 in which the grouping at 3 "N (Rl R2) is in position ~ (2-deoxy DArabino series) or c ~ (2-deoxy D ribo series) relative to the cycle, Rl and R2 identical or dirr ~ represents a hydrogen atom, a C1 to C6 alkyl group, which can form a cycle, this cycle possibly comprising a heteroatom such as an oxygen or a nitrogen, a group ~ mino ~ C1 to C6 alkyl or cyanomethyl X and Y can be the same or different and represent an OH, CH3, CH2-NH ~, X and Y can also be linked and constitute a cycle, for example, a

2-methyl 1,3 dioxane, thus forming a 4,6- type osidic bicyclic skeleton ethylidene 3-amino-2,3-dideoxy-, BD arabino or ribo-hexo pyranoside.
It also relates to their addition salts with mineral acids or organic lifi ~ nt the ote or atoms of ote, in particular hydrochlorides Advantageously, the NRlR2 group is an NH2 group or N (CH3) 2 The NRlR2 group can eg ~ lement be a substituted amino group one or two times with Ull methyl, CH2CN, CH2-CH2-NH2, form a cycle like morpholine Advantageously, the compounds of general formula I will be chosen with a glycoside for which X and Y form a cycle with an enrh ~ înrment OCH (CH3) 0CH2, such as 4,6 ethylidene-3-amino-2,3-dideoxy-, BD arabino-hexo pyranoside or 4.6 ethylidene-3-amino-2,3-dldesoxy- ~ -D-ribo-hexopyranoside.

WO 97/13776 PCT ~ FR96 / 01588 In particular, the compounds according to the invention are selected from the following compounds:
~ 4'-Demethyl-4-0 (3-amino-4,6-ethylidene-2,3-dideoxy-13-D-arabinohexopyranosyl) epipodophyllotoxin, ~ 4'-Demethyl-4-0 (3-amino-4,6-ethylidene-2,3-dideoxy- ~ -D-ribohexopyranosyl) epipodophyllotoxin, 4'-Demethyl-4-0 (3-dimethylamino-4,6-ethylidene-2,3-dideoxy-, BD-arabinoh ~ co-pyranosyl) epipodophyllotoxin, ~ 4'-Demethyl-4-0 (3-dimethylamino-4,6-ethylidene-2,3-dideoxy- ~ -D-ribohexo-pyranosyl) epipodophyllotoxin, 4'-Demethyl-4-0 (3-cyanomethylarnino-4,6-ethylidene-2,3-dideoxy- ~ -D-ribohexo-pyranosyl) epipodophyllotoxin, 4'-Demethyl-4-0 (3- (N-morpholino) -4,6-ethylidene-2,3-dideoxy- ~ -D-ribohexo-pyranosyl) epipodophyllotoxin, ~ 4'-Demethyl-4-0 [3 (2-aminoethylamino) -4,6-ethylidene-2,3-dideoxy-, BD-ribohexo-pyranosyl)] epipodophyllotoxin, 4'-Demethyl-4-0 (3-amino-2,3,6-tridesoxy- ~ -D-ribohexopyranosyl) epipodophyllotoxin, ~ 4'-Demethyl-4-0 (3,6-diamino-2,3,6-tridesoxy-, BD-ribohexopyranosyl) epipodophyllotoxin.
The ~ .eselIte invention also relates to pharmaceutical compositions comprising at least one compound of general formula I according to the invention and an excipient a ~ p.oylié.
The pharmaceutical compositions can be suitably presented by 25 ~ tlmini ~ tration by injection or orally in the form of capsules, capsules, of co ~ hllés at a dosage of I to 200 mg / m2 by injectable route and from 5 to 500 mg / m2 orally in a 24 hour period.
These derivatives can thus be ~ lmini ~ very clin hum ~ ine to treat different forms of cancer such as small cell lung cancer, cancer of 30 testes, embryonic tumors, neuroblastomas, kidney cancer, CA 02234484 l998-04-08 WO 97/13776 PCT ~ R96 / 01588 Hodgkin's and non-Hodgkin's Iphomas, acute leukemia, cancers colorectau. ~ ;, melanomas, placental choriocarcinomas and adenocarcinomas . ~ rn m ~ ires.
The present invention also relates to the procedures for preparing S compounds of formula I as well as their addition salts with mineral acids :; or pharmaceutically acceptable organic.
The present invention therefore relates to the processes for preparing the compounds of general formula I according to the invention, in which a compound of formula III or IV or V

~ '' o ~~, ~ - ji + ~~~~~ Fi + P o ~ oFi 0 lll IV V
with 4'-Demethyl 4'-benzyloxycarbonyl épipodophylloto: ~ ine with the etherate of ~ 3, or trimethyl silyl trifluorometh ~ neslllforate in an inert solvent at low temperature;
in formula III and IV, the substituent in position 3 can be ~ or ~, NRIR ~
can be an amino protected by a group Z, in formula V, P represents a protective group of alcohol and the products resulting from this condensation are deprotected and hydrogenated to provide the compounds of formula 1, the primary amines in position 3 of glycosyl are methylated by formalin and sodium cyanoborohydride.
The intermediate of formula IV is prepared by reacting a mixture of diacetoxy azido glycoside VI

Co Co N ~
~, ~ Vl CA 02234484 l998-04-08 WO 97/1 ~ 776 PCT ~ FR96 / OlS88 with tert-butyl dimethylsilyl chloride in the presence of imidazole, in that it is separated the products resulting from this reaction, in that each of these products are acetylated, 4,6-ethylidene cyclises with acetaldehyde acetal in catalytic acid medium Other characteristics of the process according to the invention will appear in the light of the description which follows, notably the process of synthesis reported in diagram I.

Ac ~
Ac (~ LI) H20 80 ~ C AcO ~
AcO ~ g 2) N ~ N3 / AcOH AcO ~ OH

N ~; ~
TsDMSiCI / Imid; ~ olc /

Co AcO ~ ~ OSi + AcO ~ j_ OS ~
~, 4 3 oll s ~ - ~ osl ~

~ IcO
~ T.
C ~ 13CIl (OE.) 2 OR, ~ r rs' Cl 13C ~ i R, T, ~ 12 CJrd ~ 0- ~. ~ ~~ OS ~
A ~ OEt. ~ .E ~ 3 ~ 6 zcl / NEt3 ~ /
ZN ~ OS ~ c1l2cl2 ~ 0 ~ = o1s ~ f 1) N ~ u ~ F, TIIF

2) BF3.E ~ 20 DMErT ~ oz WO 97113776 PCTM ~ R96 / 01588 -li2 CtPd 10 '~.
~~ 0 ~ "O ~ cOEt, NEt3 ~~ 0, ~ _ D. ~ 1ErT 10 D. ~ 1 IrT
I) llCI1O
Nai3113CN ' Cl ItC12 ~~ 0 2) I HERE ~ lrOH 1 1 D ~ r ~ loE

4 ~~ osl ~ ~~ ~
~ J 12 Clt3CN N ~ 13 A ~, '', T. '~ tSOTt T ~ set ~ 1 ~
D: ~ IEPT ~ '-0 ~ 0 Cli2C12 J 14 DhlEPT
D ~ l EPT ~~ '-OZ
112 Urd 10 ~ /.
EtOH-AcOEI

~ f ~ o ~~ oc ~ c ~ 1 f ~ N ~ D. ~ IEPT IMI NE13 / D. ~ lF ID ~ 1EPT
o 17 16 OH
O (C} 12 C} 12 1) 2 /
C} 12CI ' RT.
IC ~ 12Ci-i2NHZ
D; ~ lF Na8H3CN
C} 13CN
RT

'- H2 C / Pd ~ f ~~ 0 ~ 0 ~ cOEl-EtOll ~ O ~
o ~ ~ fo ~ or ~ N 20 4'-OH

~ - ~ ~ 4'-OH Z ~. ~} iN 'D4h ~ oE ~ PT

CA 02234484 l998-04-08 WO 97/13776 PCT ~ FR96 / 01588 It is done according to a described methodology (JC Florent and C. Monneret, J.
Chem. Soc. Chem. Comm. 1987, 1171 and B. Abbaci, JC Florent and C. Monneret, Bull.
Soc. Chim. Fr. 1989, 667) from glucal 1. The azide ion is condensed to provide a glycosidic intermediate 2 whose anomeric OH is protected by a group silyl exclusively in position, B according to the technique described (C. Kolar and G. Kneissl Angew. Chem. Int. Ed 29, 809 (1990)) to provide the mixture of the 2 epimeric azides: 3 and 4 chromatographically separable at this stage. By basic deacetylation in presence of sodium methylate, the diols in position 4, 6 are obtained: compounds 5 and 12. The azide diol compound ~ 5 is cyclized to ethylidene conventionally using acetal acetaldehyde in acid catalysis to lead to compound 6, the azide of which is reduced to amine 7 to be protected by a benzyloxycarbonyl group (Z) in compound 8, necessary for coupling with demethyl epipodophyllotoxin, itself protected on its 4 'phenol by a benzyloxycarbonyl group, this intermediate will called DMEPT4'-OZ. This coupling is carried out initially by cleavage of the protective silyl by F-ions followed by treatment with BF3 etherate at low temperature in the same medium. Deprotection of Z groups by hydrogenolysis provides compound 10 of general formula I (NRlR2 = ~ NH2; XY =
OCH (CH3) OCH2). Compound 11 of general formula I (NRIR2 = ~ NMe2; XY =
OCH (CH3) 0CH7) is obtained by methylation of the preceding primary amine by action formalin and sodium cyanoborohydride.
The glycosidic intermediary ~ ide a 12 follows the same reaction sequence as its epimerically and provides identical coupling derivative with DMEPT4'0Z from from azide 3 "to 13, in this case the coupling is carried out more easily according to two techniques.
The first technique consists in treating the ribohexopyranoside 13 derivative with trimethyl silyl trifluoromethane sulfonate (TMSOTf) at -40 ~ C in CH2Cl2. The second technique consists in using the etherate of BF3 in CH2C12 at - 15 ~ C.
The intern ~ ire obtained 14 is then catalytically reduced to lead to 15 corresponding to the general formula I where NR1R2 = aNH2 and XY = OCH (CH3) OCH2.
The same methylation as for the compound, ~ NH2 10, with formalin and CA 02234484 l998-04-08 WO 97/13776 PCT ~ R96 / 01588 cyanoborohydlul ~ leads to the corresponding dimethylamino derivative: general formula I
(NR1R2 = aNMe2; XY = OCH (CH3) 0CH2). From the primary amine lS, it is possible to alkylate nitrogen with a halogenated derivative such as iodoacetonitrile in weakly basic conditions with triethylamine in DMF and lead to the derivative S 16 of general formula I where NR1R2 = aNHCH2CN; XY = OCH (CH3) 0CH2 From same way, derivative 17 is obtained by forming the morpholine cycle by cyclization of ether diiodé on the same primary amine intermediary 15 The same al ~ ylation with the iodoethylamine protected by a group Z on the compound 15 provides the derivative 18 which is catalytically reduced to 19 derivative ~ i ~ rnino corresponding to the general formula 10 I with NR1R2 = a-NHCH2CH2NH2; XY = OCH (CH3) 0CH2.
The compounds of formula I where XY does not form a ring are obtained in the way following, ~. shown on diagram 2:

CA 02234484 l998-04-08 Ol ~ OT ~

Ho ~~ 5 ~ ¦ T Cl / P idi ~ ~ osl N ~ 12 CH2C12 N3 21 N 22 Pvridine N ~ AcO ~ O RMTNF3 ClAc ~ DMEPT4'-OZ os!
N ~ 27 DMEPT
4 OZ ~ 24 4'-OZ ~ 23 Annberlite410 \ H2C12 / l ~ -eOH

N3 \ ~ \
110 ~ 0 I ~ O-- ~ O AcOÉt HO-- ~ O

112 C / Pd NH26 ~ '-OH 25 ~ -oz NEt3 HO ~, ~ _ O
J''1} 2 DMEPT
OH

the silylated azide 12 is selectively tosylated with tosyl chloride in pyridine in 21, the primary alcohol tosylate is exchanged for the iodo derivative 22. At this stage, the alcohol Secondary 5 in position 4 is protected by a chloroacetate to provide 2-deoxysugar operates 23 ready to be condensed with DMEPT 4'-OZ under the conditions usual with BF3 etherate in methylene chloride at low temperature.
Intermediate 24 obtained, allows by passage over the amberlite resin IRA410 treated in basic medium to protect the chloroacetate group from compound 25 and the hydrogenation 10 final catalytic allows in a state to reduce the azide function in arnino 3 "a, unprotect the Z function in position 4 'and reduce the 6 "carbon to methyl for provide the drift 26 corresponding to the general formula I where NR1R2 = 3 "aNH2, X =

CA 02234484 l998-04-08 W 0 97/13776 PCT ~ R96 / 01588 OH; Y = CH3. Intermediate 24 obtained previously can react with the azide ion in the DMF at telllLJel ~ L ~ ambient to supply the 3 "aN3, 6" -N3 co ~ olL ~ lt un azidoacetate at position 4 "27, which can be cut into alcohol 4" by a treatment similar to the previous one: switching to Amberlite IRA 410 exchange resin for S provide the rli ~ 7.i (1o ~ lcool 28. The final catalytic reduction step makes it possible to obtain the derivative di ~ miné 29 corresponding to the general formula I (where NR1R2 = 3 "aNH2; X =
OH, Y = CH2NH2). The salts formed from the nitrogen compounds are, for example, hydrochlorides and are formed conventionally by treating a methanolic solution of the nitrogen compound with a stoichiometric solution relative to the sites to be salified of m ~ th ~ nl hydrochloric acid previously dosed. The crystallized hydrochloride can optionally obtained by precipitation in the reaction medium by addition of ether ethyl.
For information and to illustrate the different stages of the synthesis, but to without limitation, the following examples are given.
F ~ FMPr.F 1 General formula I: NRlR2 =, ~ NH2; XY = OCH (CH3) OCH2 4'-Demethyl-4-0 (3-amino-2,3-dideoxy-4,6-ethylidene- ~ -D-arabinohexopyranosyl) epipodophyllotoxin (compound 10) Stage I
A solution of tri-O-acetyl-D-glucal I (50 g; 183 mmol) in water (400 ml) is heated for 3 h at 80 ~ C. The reaction medium is then cooled to 20 ~ C before addition of sodium azide (17.9 g; 275 mmol) and acetic acid (38 ml;
600 mrnol) After stirring at room temperature for 24 h, the medium is neutralized with NaHCO3 (salt). The phase is extracted with ethyl acetate (3 x 500 ml). The organic phases are combined, dried over MgSO4 and then concentrated under reduced pressure. This provides 51 g of crude product 2 immediately treated as follows.
Characteristics: CCM: cyclohexane / AcOEt: 1/1; Rf = 0.43 CloH1sN3O6 M = 273 CA 02234484 l998-04-08 WO 97/13776 PCT ~ FR96 / 01588 St ~
Tertbutyldimethylsilyl-3 -azido-2,3-dideoxy-4,6-di-O-acetyl-, BD-arabino-hexopyranoside 3 and terbutyldimethylsilyl-3-azido-2,3-dideo ~ {y-4,6-di-O-acetyl-, BD-ribohexopyranoside 4 Aunesolution dumange brut2 (16.1 g; 58mmol) obtaineduustust I in anhydrous dichloromethane (200 ml) previously cooled to 0 ~ C, are added successively, under argon, 6 g of imidazole (87.8 mmol) and 13.24 g of chloride tertbutyldimethylsilyl (87.8 mmol). After stirring for 15 min at 0 ~ C and 19 h at 20 ~ C, the reaction medium is poured into 500 ml H20. The aqueous phase is ex ~ aite with CH2C12 (200 ml) then after drying over MgSO4, the organic phase is concentrated under reduced pressure and the residue (18.7 g) is chromatographed on silica (cyclohexane / AcOEt: 9/1). 10.7 g of 3 are thus isolated (syrup; 48%) and 4.7 g of 4 (syrup; 21%); however that intermediate fractions contain a mixture of 3 and 4 (3.3 g; 15%).
Characteristics: 3 CCM: cyclohexane / AcOEt: 4/1 Rf = 0.45 [a] D20 = - 10 ~ (c = 1.4; CHC13) MS: m / z 405 (M + NH4) C16H29N3O6Si M = 387 4 Rf = 0.52 [a] D20 = + 10 ~ (c = l; CHC13) MS: mJz 405 (M + NH4) +
C16H2gN3O6Si M = 387 lH NMR 300MHz CDC13 ~
Derivative 3: 0.13 (3H, s, SiCH3); 0.14 (3H, s SiCH3);
0.92 (9H, s, tBu); 1.72 (1H, m, J2a-1 = 9 ~ 5 Hz ~
J2a-2e = 12.5 Hz, J2a-3 = 12.5 Hz, H2a) 2.05 (3H, s, COCH3); 2.15 (3H, s, COCH3); 2.25 (1H, ddd, J2e-1 = 1.5 Hz, J2e-2a = 12 ~ 5 Hz ~ J2e-3 = 4.5 Hz, H2e); 3.55 - 3.62 (2H, m, H3 and Hs);

4.10 (1H, dd, J6-s = 2.5 Hz, H6);
4.20 (1H, dd, J6-s = 6 Hz, J6-6 ~ = 12 Hz, H6 ');
4.86 (1H, t, J = 9.5 Hz, H4);
4.86 (1H, dd, Jl-2a = 9.5 Hz, Jl-2e = 1.5 Hz ~ Hl) s Derivative 4: 0.1 (6H, s, Si (CH3) 2); 0.88 (9H, s, tBu);
1.64 (IH, ddd, J2a-2e = 14 Hz, J2a-l = 8.5 Hz, J2a-3 = 3.5 Hz, H2a);
2.03 (1H, ddd, J2e-2a = 14 Hz, J2e-1 = 2 Hz, J2e-3 = 4 Hz, H2e);
2.05 (3H, s, COCH3); 2.13 (3H, s, COCH3);
4.05 -4.12 (1H, m, Hs);
4.17 - 4.22 (3H, m, H3, H6 and H6); 4.89 (1 H, dd, J4 5 = 9.5 Hz, J4 3 = 3.5 Hz, H4);
5.02 (1H, dd, Jl-2a = 8.5Hz, Jl-2e = 2Hz, Hl).
Stage 3 Tertbutyldimethylsilyl-3-azido-2,3-dideoxy-, ~ -D-arabino-hexopyranoside S
To a solution of derivative 3 (3 g; 7.7 mmol) obtained in stage 2 in anhydrous methanol (40 ml) is added, under argon, a 1M sodium methanolate solution (1.9 ml). After reaction for 1 h 30 at 20 ~ C, the reaction medium is brought back to pH = 7 by addition of H + residue (AmberliteTM IRC 50 S). The mixture the reaction mixture is filtered and the filtrate concentrated under reduced pressure leading to 2.27 g of 5 (97%).
Characteristics: TLC: cyclohexane / AcOEt: 2/1; Rf = 0.36 [(x] D20 = - 26 ~ (c = 1; CHC13) MS: m / z 304 (M + H) + 321 (M + NH4) +
Pf = 70 - 72 ~ C
C12H2sN304Si M = 303 Sta ~ e 4 Tertbutyldimethylsilyl-3-azido-2,3-dideoxy-4,6-O-ethylidene-, BD-arabino-hexopyranoside (6) To a solution of 5 (0.20 g; 0.6 mmol) obtained in stage 3 in 5 ml ~ U ~ ~~ IF ~~ E

CA 02234484 l998-04-08 WO 97/13776 PCT ~ R96 / 01588 14 of acetonitrile, 0.94 ml (6.6 mmol) of the acetaldehyde diethylacetal is added and then 15 mg paratoluenesulfonic acid (0.08 mmol). The reaction medium is stirred at room temperature for 1 h then diluted with ethyl acetate (~ 0 ml) before washing with a sodium hydrogencarbonate solution (pH = 9) (20 ml) then with water 5 (20 ml). The organic phase is dried over MgSO4 and then concentrated under reduced pressure to provide 0.25 g of crude product. Purification on silica gel (cyclohexane / AcOEt: 9515) allows 0.19 g of pure 6 (B6%) to be isolated.
Characteristics: TLC: cyclohexane / AcOEt: 7/3; Rf = 0.89 r ~] D20 = - 19 ~ (CI, 1, CHC13) MS: m / z 347 (M + NH4) +
C14H27N304 If M = 329 CHN
Calculated 5 1.04 8.26 12.75 Found 51.64 8.43 12.51 15 Stadium ~
Tertbutyldimethylsilyl-3 -amino-2,3-dideoxy-4,6-0-ethylidene- ~ -D-arabino-hexopyranoside (7) To a solution of 6 (2 g; 6 mmol) obtained in stage 4 in 30 ml of acetate ethyl are added 50 ~ 11 of triethylamine then 0.5 g of 10% palladium on carbon. The ~ 0 reaction medium is placed under a hydrogen atmosphere (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 provide 1.82 g of pure 7 (98%) ~
Characteristics: CCM: cyclohexane / AcOEt: 1/1; Rf = 0.23 [a ~ D20 = - 28 ~ (C = 1.3; CHCI3) SM: ml'z 304 (M + H) +
C14H2gN04Si M = 303 St ~ -le 6 Tertbutyldimethylsilyl-3 -aminobenzyloxycarbonyl-2,3-dideoxy-4,6-0-ethylidene- ~ -D-30 arabino-hexopyranoside (8) -WO 97/13776 PCT ~ FR96 / 01588 Benzyloxycarbonyl chloride (1.17 ml; 7.88 mmol) is added, under argon, to a solution previously cooled to 0 ~ C of acetal 7 (1.82 g; 6 mmol) obtained at stage ~ in a mixture of dichloromethane (30 ml) and triethylamine (1.27 ml; 9.1 mmol) anhydrous. After stirring for 8 h, the reaction medium is poured into 100 ml - 5 H2O and the aqueous phase is extracted with CH2C12 (100 ml). The organic phase is dried over MgSO4, concentrated under reduced pressure and the residue is purified by chromatography on silica gel (cyclohexane / AcOEt: 6/1 and 4/1) in order to isolate 1.9 g of 8 (72%).
Characteristics: CCM: cyclohexane / AcOEt: 1/1; Rf = 0.64 [a] D20 = - 27 ~ (c = 1.14; CHC13) MS: m / z 438 (M + H) +
Pf = 102 ~ C
C22H35No6si M = 437 CHN
Calculated 60.38 8.06 3.20 Found 60.27 8.10 3.29 Stage 7

3-aminobenzyloxycarbonyl-2,3-dideoxy-4,6-O-ethylidene- ~ -D-arabino-4'-benzyloxycarbonyl-epipodophylloto hexine xranoside ~ cine (9) 8 sugar (2.0 g; 4.57 mlnol) obtained in stage 6 in solution in anhydrous dichloromethane (100 ml) are added 5.06 ml of tetra fluoride butylammoniurn (1.1 M solution in THF; 5.5 mmol). When the total disappearance of 8 is found by TLC (2 h of stirring), the reaction medium is cooled to - 20 ~ C.
DMEPT 4'-OZ (2.57 g; 4.8 mmol) is then added successively then 8.44 ml of BF3.Et2O (68.6 mmol). After reaction for 1 h at -20 ~ C, the reaction medium is pour into 200 ml of a saturated NaHCO3 solution (addition of NaHCO3 salts) (pH = 9). The organic phase is dried over MgSO4, concentrated under reduced pressure, then the crude residue (6.2 g) is chromatographed on silica gel (CH2Cl2 / Acetone: 98/2 then 97/3) to provide 9 (2.1 g; 54%).
-- WO 97/13776 PCT ~ FR96 / OlS88 Characteristics: CCM: CH2CI ~ / Acetone: 92/8; Rf = 0.61 [(x] D20 = - 74 ~ (c = 1.1; CHC13) MS: m / z857 (M + NH4) +
Pf = 175 ~ C
C4sH4sNO 15 M = 839 CHN
Calculated 64.36 5.40 1.67 Found 64.21 5.30 1.58 StZlllt? 8 10A a solution of 9 (0.28 g; 0.33 mmol) in 20 ml of ethyl acetate are added 30! 11 of triethylamine then 150 mg of 10% palladium on carbon. The middle reaction is placed under a hydrogen atmosphere (atmospheric pressure). After stirring for 1 h 30 at room temperature, the catalyst is removed by filtration and the organic phase concentrated under reduced pressure and then chromatographed on 15silica (CH2C12 / MeOH: 97/3 then 95/5) to provide ~ ir 172 mg of pure compound 10 (90%).
(Recri ~ t ~ llic ~ tion in CH2C12 / pentane).
Characteristics: TLC: CH2C12 / MeOH: 95/5; Rf = 0.31 [a] D20 = - 120 ~ (c = 1.05; CHC13) SM: ITI / Z 594 (M + Na) + 610 (M + K) +
Pf = 21g ~ C
C2gH33NO1 1 M = 571 CHN
Calculated 60.94 5.82 2.45 Found 60.45 5.78 2.58 1H NMR 300 MHz CDCl3 ~ 1.36 (3H, d, J = SHz, CH3-CH); 1.51 (1H, m, H2 ~ a);
2.05 (1H, m, H2 ~ e); 2.88 (1H, m, H3); 3.02 (2H, m, H3 ~ and H4 ~); 3.28 (1H, m, J2-1 = 5.2 Hz, H2); 3.30 (1H, m, Hs ~); 3.57 (1H, t, J = 10 Hz, H6 ~ a), 3.75 (6H, s, OCH3); 4.15 (1H, dd, J = S Hz, J = 10 Hz, H6 ~ e); 4.41 (1H, dd, J = 9 Hz, Hl 1a); 4.21 (1H, t, J = 9Hz, Hllb); 4.59 (1H, d, J = 5.2Hz, Hl); 4.75 (1H, q, J = SHz, H7 ~); 4.85 (1H, dd, J = 9 Hz, J = 2 Hz, Hl ~); 4.94 (1H, d, J = 3.3 Hz, H4); 5.98 (1H, d, WO 97/13776 PCT ~ FR96 / 01588 OCHAO); 6.00 (1H, d, OCHgO); 6.24 (2H, s, H ~ and H6 ~); 6.55 (1H, s, Hg); 6.75 (1H, s, H5).
- Preparation of chlorl ~ ydrate Amine 10 (61 mg; 0.10 mmol) dissolved in anhydrous dichloromethane (6 ml) is added a 0.098 M hydrochloric methanol solution (1.09 ml;
0.106 mmol). The reaction medium is stirred for 10 minutt-s The expected product is precipitated after addition of ether (20 ml). 56 mg (86%) of the hydrochloride are collected.
10.
Car ~ cterictiques: Pf = 230 ~ C
C29H32N ~ 1 lCl M = 606 Solubility test: 2.56 mg in 0.3 ml of water C = 0.014 M
F ~ FMPT F 2 General formula I (NR1R2 = ~ BNMe2; XY = OCH (CH3) OCH2)

4'-Demethyl-4-0 (3-dimethylarnino-2,3-didesoxy-4,6-ethylidene-, BD-arabinohexopyranosyl) epipodophyllotoxin (Compound 11) To a solution of 10 (0.19 g; 0.33 mmol) in anhydrous dichloromethane (15 ml) are added successively formaldehyde (13.5 ~ 11) and cyanoborohydride sodium (85 mg). After stirring for 45 min at ambient temperature, these same 20 reagents are added and the reaction is continued for 45 min. The middle reaction is diluted with CH2Cl2 (30 ml) and washed with water (40 ml). The sentence organic is dried over MgSO4 and concentrated under reduced pressure. The residue is chromatographed on silica gel (CH2C12 / MeOH: 97/3). This provides 101 mg of 11 (51%).
Characteristics: TLC: CH2C12 / MeOH: 95/5; Rf = 0.4 [a] D20 = -121 ~ (c = 1; CHC13) SM: m / z 600 (M + H) +
Pf = 270 ~ C
C31H37NOll M 599 1HRMN 300 MHz CDC13 ~ 1.38 (3H, d, J = 5 Hz, CH3 - CH); 1.55 (1H, m, H2 ~ a);

CA 02234484 l998-04-08 WO 97/13776 PCT ~ FR96 / 01588 1.96 (1H, m, H2 ~ a); 1.96 (1H, m, H2 ~ e); 2.33 (3H, s, CH3 - N); 2.91 - 2.82 (1H, m, H3); 2.91 - 2.82 (1H, m, H3 ~); 3.35 - 3.25 (1H, m, Hs ~); 3.38 (1H, t, J4 ~ 5 ~ = 9 Hz, J4 ~ 3 ~ = 9 Hz, H4); 3.58 (1H, t, J6 ~ as ~ = 10 Hz, J6 ~ a-6 ~ e = 10 Hz, H6 "a); 3.75 (3H ~
s, CH3 O); 4.16 (1H, dd, J6 ~ e-5 ~ = 5 Hz, J6 ~ e-6 ~ a = 10 Hz, H6 ~ e); 4.21 (1H, t, Jgb ga = 9 Hz, Jgb 3 = 8 Hz, Hgb); 4.42 (1H, dd, Jga gb = 9 Hz, Jga 3 = 10.5 Hz, Hga); 4.59 (1H, d, J1-2 = 5.2Hz, H1); 4.74 (1H, q, J = 5 Hz, CH -CH3); 4.82 (1H, dd, Jl ~ -2 ~ a = 9.5 Hz, Jl ~ -2 ~ e = 2 Hz, Hl ~); 4.95 (1H, d, J4 3 = 3.2 Hz, H4); 5.97 (1H, d, OCHAO); 6.00 (1H, d, OCHgO); 6.25 (2H, s, H2, and H6 ~); 6.55 (1H, s, Hg); 6.76 (1H, s, Hs).
10 Pre ~ r ~ tion of chlorhvdrate With amine 11 (101 mg; 0.17 mmol) dissolved in dichloromethane anhydrous (7 ml) is added a 0.098 M hydrochloric methanol solution (1.72 ml;
0.17 mrnol). The reaction medium is stirred for 10 min ~ ltP ~. The expected product is precipitated after addition of ether (20 ml). 86 mg (81%) of the hydrochloride is collected.
15 11.
Characteristics: Pf = 199 ~ C
C3 1 H33NO 1 1 Cl M = 635 Solubility test: 2.5 mg in 0.1 ml of water C = 0.03 ~ M
F.XFMPT.F 3 General formula I: NRlR2 = a-NH2; XY = OCH (CH3) OCH2 4'-Demethyl-4-0- (3-amino-2,3-dideoxy-4,6-ethylidene ~ -D-ribohexopyranosyl) epipodophyllotoxin (compound 19) St ~ e 1 Tertbutyldimethylsilyl-3-azido-2,3-didesoxy-, BD ribo hexopyranoside (compound 12) Similarly, in stage 3 of example l, but in tltili ~ nt compound 4, we obtains compound 12 which is engaged dile., ~ e, llent in stage 2.
st ~ e 2 Tertbutyldimethylsilyl-3-azido-2,3-dideoxy-4,6-O-ethylidene-, BD-ribo hexopyranoside (13) CA 02234484 l998-04-08 WO 97/13776 PCT ~ FR96 / 01588 19 To a solution of 12 (0.25 g; 0.8 mmol) obtained in stage 1 in 10 ml of acetonitrile, 1.1 ml (8 mmol) of the acetaldehyde diethylacetal and then 52 mg are added - C ~ mphorsulfonic acid (0.24 mmol ~. The reaction medium is stirred at temperature ambient for 9 h then diluted with ethyl acetate (30 ml) before washing with a

5 sodium hydrogencarbonate solution (pH = 9) then with water (30 ml). The sentence organic is dried over MgS04 then concentrated under reduced pressure to provide 0.3 g of crude product. Purification on silica gel (cyclohexane / AcOEt: 15 µl) isolates 0.15 g of pure 13 (55%).
Characteristics: CCM: cyclohe ~ cane / AcOEt: 4/1; Rf = 0.77 [a] D20 = - 35 ~ (c = I, 1 CHC13) C14H27N3O4Si M = 379 Stage 3 3-azido-2,3-dideoxy-4,6-O-ethylidene- ~ -D-ribo-hexopyranoside from 4'-ben2yloxycarbonyl-epipodophyllotixne (compound 14) 15 st synthetic route:
To a mixture of DMEPT 4'-OZ (438 mg; 0.82 mmol), 13 (270 mg;
0.82 mmol) obtained in stage 2 and 4 A molecular sieve (1.5 g) in anhydrous dichloromethane (30 ml) cooled to -40 ~ C, is added trimethylsilyl-trifluorometh ~ n ~ sl ~ lfonate (TMSOTf) (446! 11; 2.46 mmol). After reaction during I h 15 to - 40 ~ C, the reaction medium is neutralized with triethylamine (347 ~ 1), filtered then washed with saturated NaCl solution (20 ml). The organic phase is dried over MgSO4, concentrated under reduced pressure, then the crude residue is chromatographed on silica gel (cyclohexane / AcOEt: 65/35) to provide 14 (260 mg; 45%).
2nd synthetic route:
To a mixture of DMEPT 4'0Z (1.85 g; 3.46 mmol), of 13 (1.20 g; 3.64 mmol) obtained in stage 2 in anhydrous dichlorometh ~ (100 ml) cooled to -15 ~ C, is a ~ outé boron trifluoride etherate (BF3.Et2O) (425 ~ Ll; 3.46 mmol). After reaction for 2 h at - 15 ~ C, the reaction medium is diluted with 100 ml of C ~ I2C12 then, poured in 200 ml of a saturated NaHCO3 solution. The organic phase is dried over MgSO4, concentrated under reduced pressure then, the crude residue (2.9 g) is WO 97/13776 PCT ~ FR96 / 01588 chromatographed on silica gel (cyclohe ~ cane / AcOEt: 65/35) to provide 14 (1.19 g;
47%) (Et20 / hexane recrystallization).
Characteristics: CCM: cyclohe.Yane / AcOEt: 6/4; Rf = 0.41 cyclohe ~ ane / AcOEt: 65/3; Rf = 0.77 [a] D20 = - 105 ~ (c = 1.05; CHC13) MS: m / z 749 (M + NH4) +
Pf = 139 ~ C
C37H37N3O 1 3 M = 731 St ~ 4 3-amino-2,3-dideoxy-4,6-O-ethylidene- ~ -D- ~ Libo-hexopyranoside epipodophyllotixin 15 To a solution of 14 (110 mg; 0.15 mmol) obtained in stage 3 in a mixture ~ 10 ml of ethanol and 5 ml of ethyl acetate are added 20 ~ LI of triethylamine then 20 mg of 10% palladium on carbon. The reaction medium is placed under an atmosphere 15 of hydrogen (atmospheric pressure). After stirring for 2 h at temperature ambient, the catalyst is removed by filtration and the organic phase concentrated under reduced pressure then chromatographed on silica gel (CH2C12 / MeOH: 97/3 then 95/5) to fo ~ 3rnir 63 mg of pure 15 (7 2%).
Characteristics: CCM: CH ~ CI ~ / MeOH: 95/5; Rf = 0.39 [a] D ~ 0 = - 100 ~ (c = 1.05; CHC13) MS: m / z 572 (M + 1) 589 (M + NH4) +
Pf = 217 ~ C
C2gH33NO1 1 M = 571 1 HRMN 300 MHz CDC13 o: 1.35 (3H, d, J = 5 Hz, CH3-CH); 1.73 (1H, m, H2 ~ a);
1.90 (1H, m, H2 ~ e); 2.83 (1H, m, H3); 3.22 (1H, dd, J2-1 = 5.2 Hz, J2-3 = 14 Hz, H2);
3.42 (1H, dd, J4 ~ 3 ~ = 9.5 Hz, J4 ~ 5 ~ = 3.5 Hz, H4 ~); 3.74 (6H, s, OCH3); 3.49 - 3.60 (2H, m, H3 ~ and H6 ~); 3.94 - 4.02 (1H, m, Hs ~); 4.10 - 4.20 (2H, m, H6 ~ and Hl lb); 4.42 (1H ~ dd ~ Jlla-3 = 9Hz, Jlla-llb = 9.5 Hz, Hlla); 4.57 (1H, d, Jl-2 = 5.2Hz, Hl);
4.78 (1H, q, J = 5Hz, H7 ~); 4.91 (1H, d, J43 = 3.4Hz, H4); 5.38 (1H, dd, J1 ~ 2 ~ e = 2 Hz, Jl ~ -2 ~ a = 9 Hz, Hl ~); 5.93 l ~ 1H, s, OCHAO); 5.97 (1H, s, OCHgO); 6.25 (2Ht s, CA 02234484 l998-04-08 WO 97/13776 ~ CT ~ R96 / 01588 H2 ~ and H6 ~); 6.51 (1H, s, Hg); 6.86 (1H, s, Hs).
Preparation of the hydrochloride - At the amine lS (50 mg; 0.087 mmol) obtained in stage 4 are added ~ 890, ul of a 0.098 M hydrochloric acid solution (0.087 mmol). After shaking for 10 min ~ ltec and addition of ether (10 ml), the crystals or hydrochloride (52 mg, 98%) are obtained by filtration.
Characteristics: Pf = 1 75 ~ C
C29H34N ~ l 1 Cl M = 607 Solubility test: 2.2 mg in 0.2 ml of water C = 0.02 M
F ~ .MPr F. 4 General formula I: NR1R2 = a-NMe2; XY = OCH (CH3) OCH2 4'-Demethyl-4-0 (3 -N, N-dimethylamino-2,3-dideoxy-4,6-O-ethylidene- ~ -D-ribo-hexopyranosyl) epipodophyllotoxin 20 To a solution of lS (29 mg: 0.05 mmol) obtained in stage 4 of Example 3 in ml of acetonitrile are added successively formaldehyde (10.3 ~ ll) and sodium cyanoborohydride (12 mg). After stirring for 2 h at temperature ambient, the reaction medium is diluted with CH2Cl2 (20 ml) and washed with water (20 ml). The organic phase is dried over MgSO4 and concentrated under reduced pressure.
The residue is reacted under the same conditions and after treatment identical chromatographed on silica gel (CH ~ Cl2 / MeOH: 97/3). This provides 29 mg of 20 (95%).
Characteristics: TLC: CH2Cl2 / MeOH: 95/5; Rf = 0.5 [a] D20 = - 85 ~ (c = 1.06; CHCl3) SM: m / z 600 (M + H) +
Pf = 140 ~ C
C31H37NOl 1 M = 599 1 H NMR 300 MHz CDC13 o: 1.35 (3H, d, J = 5 Hz); 1.55 (1H, m, H2 ~ a); 2.20 (1H, m, H2 ~ e); 2.36 (3H, s, CH3 N); 2.62 (1H, m, 2.62, H3 ~); 2.85 (1H, m, H3); 3.23 (1H, dd, J2-1 = 5.2 Hz, J2-3 = 14 Hz, H2); 3.52 (1H, dd, J4 ~ 3 = 3 Hz, J4 ~ 3 ~ = 9 Hz, H4 ~

CA 02234484 l998-04-08 WO 97/13776 PCT ~ R96 / 01588 3t61 (1H, t, J6 ~ a-6 ~ e = 10 Hz, J6 ~ a-5 ~ = 10 Hz, H6 ~ a); 3.75 (3H, s, CH3 O); 4.22 - 4.02 (1H, m, Hs ~); 4.22 - 4.02 (1H, m, H6 ~ e); 4.22 - 4.02 (1H, m, Hgb); 4.43 (IH, dd, Jga gb = 9 Hz, Jgb 3 = 10.5 Hz, Hgb); 4.62 - 4.57 (1H, m ~ Jl-2 = 5.2 Hz, Hl); 4.62 - 4.57 (3H, m, J = 5 Hz, CH-CH3), 4.88 (1H, d, J4 3 = 3.4 Hz, H4); 5.96 (1H, d, OCHA ~);
5.98 (1H, d, OCHg ~); 6.25 (2H, s, H2 and H6 ~); 6.52 (1H, s, Hg); 6.80 (1H, s, Hs).
Preparation of chlor ~ ydrate With amine 20 (59 mg; 0.1 mmol) dissolved in anhydrous methanol (2 ml) 0.098 M hydrochloric methanol solution (1 ml; 0.1 mmol) is added. The reaction medium is stirred for 10 min-t ~ s The expected product is precipitated after addition of ether (20 ml). 33 mg (53%) of crystals are collected.
Characteristics: Pf = 1 50 ~ C
C31H38N ~ l lCl M = 635 Solubility test: 2.6 mg in 0.1 ml of water C = 0.04 M
F ~ FrvIpr F. S
General formula ~: NR1R2 = ~ -NHCH2CN; XY = OCH (CH3) OCH2 4'-Demethyl-4-0- (3-cyanomethylamino-2,3-dideoxy-4,6-ethylidene-, BD-ribo-hexopyranosyl) epipodophyllotoxin (Compound 16) To a solution of 15 (120 mg; 0.21 mmol) obtained in stage 4 of Example 3 in 4 ml of dimethylformamide are added 200 ~ LI of triethylamine (1.47 mmol) then 100 ~ L of iodoacetonitrile (1.47 mmol). The reaction medium is stirred for 20 h at room temperature then diluted with ethyl acetate (30 ml) before washing with water (4 x 30 ml). The organic phase is dried over MgSO4, concentrated under pressure reduced then chromatographed on silica gel (CH2C12 / acetone: 92/8) to provide 71 mg of pure 16 (55%).
Characteristics: TLC: CH2C12 / MeOH: 95/5; Rf = 0.25 [a] D20 = - 86 ~ (c = 0.80; CHCl3) SM: m / z 611 (M + H) + 628 (M + NH4) +
C31H34N2 ~ ll M = 610 CA 02234484 l998-04-08 WO 97/13776 PCT ~ R96 / OlS88 1H NMR 300 MHz CDCl3 ~: 1.35 (3H, d, J = SHz, CH3CH); 1.69 (1H, m, J2 ~ a-1 =
9.5 Hz, J2 ~ a-2 ~ e = 13 Hz, J2 ~ a-3 = 3 Hz, H2 "a ~; 1.93 (1H, m, H2 ~ e); 2.86 (1H, m , H3); 3.25 (1H, dd, J2-1 = 5.2 Hz, J2-3 = 14 Hz, H2); 3.51 - 3.56 (2H, m, H4 ~ and H6 ~ a); 3.56 (2H, m, CH2CN); 3.76 (6H, s, OCH3); 3.88 (1H, m, Hs ~); 4.17 (2H, t, Jlla-3 = 8 Hz, Jlla-llb = 10 Hz, H3 ~ and Hllb); 4.14 (1H, dd, J6a ~ -6 ~ b = 10 Hz, J6 "b-5" = 5 Hz, H6 ~ b); 4.41 (1H, dd, Jl la-3 = 9 Hz, Jl la-1 lb = 10 Hz, Hl la); 4 ~ 60 (1H, d, Jl-2 = 5.3 Hz, Hl); 4.75 (1H, q, J = 5 Hz, H7 ~); 4.90 (1H, d, J4 3 = 3.3 Hz, H4); 5.17 (1H, dd, Jl "-2" c = 2 Hz, Jl ~ -2 ~ a = 9.5 Hz, Hl ~); 5.99 (1H, s, OCHAO);

6.00 (1H, s, OCHgO); 6.25 (2H, s, H2 and H6 ~); 6.54 (1H, s, Hg); 6.79 (1H, s, Hs).
10 F ~ F.MPLF 6 General formula I: NRlR2 = a-morpholino; XY = OCH (CH3) 0CH2 4'-Demethyl-4-0- (3 -N-morpholino-2,3-didesoxy-4,6-ethylidene-, ~ -D-ribo-hexopyranosyl) epipodophyllotoxin (Compound 17) To a solution of 15 (60 mg; 0.10 mmol) obtained in stage 4 of Example 3 in 2 ml of dimethylformamide are added 58 ~ 11 of triethylamine (0.42 mmol) then 512 mg of di-iodoethyl ether (1.57 mmol). The reaction medium is stirred for 96 h at at room temperature and in the dark, then diluted with ethyl acetate (30 ml) before washing with water (4 x 30 ml). The organic phase is dried over MgS04, concentrated under reduced pressure then chromatographed on silica gel (CH2Cl2 / acetone: 92/8) to provide 46 mg of pure 17 (68%).
Characteristics: CCM: CH2C12 / acetone: 92/8; Rf = 0.31 [a] D20 = - 98 ~ (c = 1.04; CHC13) C33H3gN012 M = 641 1H NMR 300 MHz CDCl3 ~: 1.33 (3H, d, J = SHz, CH3CH); 1.55 (1H, m, J2 ~ a-1 ~ =
9.5 Hz, J2 ~ a-2 ~ e = 13 Hz, J2 ~ a-3 ~ = 3 Hz, H2 "a); 2.15 (1H, m, H2 ~ e); 2.84-2, 90 (5H, m, CH2N and H3); 2.80 (1H, dd, J3 ~ 4 ~ = 3Hz, J3 ~ -2 ~ a = 3 Hz, J3 ~ -2 ~ e = 3 Hz, H3 ~
3.23 (1H, dd, J2-1 = 5.2Hz, J2-3 = 14Hz, H2); 3.48 (1H, t, J6 ~ a-6 ~ b = 12 Hz, J6 "as"
= 12 Hz, H6 ~ a); 3.57 (1H, dd, J3 ~ 4 ~ = 3Hz, J4 ~ 5 ~ = 9 Hz, H4 ~); 3.66-3.76 (10H, m, OCH3 and OCH2); 4.08-4.16 (2H, m, Hs ~ and H6 "b); 4.20 (1H, t, Jl la-3 = 8 Hz, Jl la-11 lb = 9 Hz, Hlla); 4.43 (1H, dd, Jllb-3 = 9 Hz, Jllb-11a = 9 Hz, H1lb); 4.57-4.62 CA 02234484 l998-04-08 WO 97/13776 PCT ~ FR96 / 01588 (2H, m, H1 and H7 ~); 4.89 (1H, d, J4 3 = 3.4 Hz, H4); 5.20 (1H, dd, Jl ~ -2 ~ e = 2 Hz, Jl ~ -2 ~ a = 9.5 Hz, Hl ~); 5.97 (1H, s, OCHAO); 6.00 (1H, s, OCHgO); 6.25 (2H, s, H2 and H6 ~); 6.54 (1H, s, Hg); 6.73 (1H, s, Hs).
F ~ FMPT E 7 General formula I: NR1R2 = a-NH2 (CH2) 2NH2; XY = OCH (CH3) OCH2 4'-Demethyl-4-0 [3- (2-Aminoethylamino) 2,3-dideoxy-4,6-ethylidene- ~ -D-ribo-hexopyranosyl] epipodophyllotoxin 18 To a solution of 15 (173 mg; 0.30 mmol) obtained in stage 4 of Example 3 in 10 ml of dimethylfom ~ are added triethylamine (127 ~ L1; 0.91 mmol) and N-benzyloxycarbonyl-2-iodoethylamine (0.28 g; 0.91 mmol). The middle reaction mixture is stirred for 5 days at room temperature then, diluted with water (30 ml). After extraction of ethyl acetate (30 ml) washing with water (5 ~ c 20 ml), the phase organic is dried over MgSO4, concentrated under reduced pressure, then chromatographed on silica gel (CH2C12 / MeOH: 97/3) to provide 155 mg of 18pur (68%).
Characteristics: TLC: CH2C12 / MeOH: 95/5; Rf = 0.70 [a] D20 = - 74 ~ (c = 1.17; CHC13) MS:} n / z 749 (M + H) +
C3gH44N2O 1 3 M = 748 which is directly involved in the following debenzylation stage:
To a solution of 18 (0.15 g; 0.20 mmol) in a mixture of ethyl acetate and ethanol (10 ml, 1/1) are added triethylamine (30, ul) and then palladium on carbon 10% (0.1 g). The reaction medium is placed under a hydrogen atmosphere (pressure atmospheric).
~ 25 After stirring for 1 h 30 at room temperature 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 silica gel (CH2C12 / MeOH (NH3): 97/3) to provide 107 mg (84%) of 19.
Characteristics: TLC: CH2C12 / MeOH (NH3): 95/5; Rf = 0.22 [a] D20 = - 77 ~ (c = 1; CHC13) CA 02234484 l998-04-08 Pf = 130 ~ C
C31H38NOl 1 M = 614 1 H Rl ~ 300 M [Hz CDC13 ~: 1.33 (3H, d, J = 5 Hz); 1.5 ~ (1H, m, H2 ~ a); 2.15 (1H, m, H2 ~ e); 2.33 (3H, m, NH2 and NH exchangeable); 2.84-2.90 (SH, m, CH2-N and H3); 3.20 (lH ~ dd ~ J3 ~ 4 ~ = 3 Hz, J3 ~ -2 ~ a = 3 Hz, J3 ~ -2 ~ e = 3 Hz, H3 ~); 3.23 (1H, dd, H2), 3.48 (lH ~ t, Jb ~ a-6 ~ b = 12 Hz, J6 "as" = 12 Hz, H6 ~ a); 3.57 (1H, dd, J4 ~ 3 ~ = 3 Hz, J4 ~ 5 ~ =
9 Hz, H4 ~); 3.75 (6H, s, OCH3); 4.08-4.16 (2H, m, Hs ~ and H6 ~ b); 4.20 (1H, t, J1 lb-3 = 8, Jl lb-l la = 9 Hz, Hl lb); 4.43 (1H, dd, Jl la-3 = 9 Hz, Jl la-1 lb = 9 Hz, Hl la ~;
4.57-4.62 (2H, m, H1 and H7 ~); 4.89 (1H, d, J4 3 = 3.4 Hz, H4); 5.20 (1H, dd, J1 ~ -2 ~ e =
2Hz, J1 ~ -2 ~ a = 9.5 Hz, H1 ~); 5.97 and 6.00 (2H, d, OCH2O); 6.25 (2H, s, H2 ~ and H6 ~);
6.54 (1H, s, Hg); 6.73 (1H, s, Hs).
Plc: p ~ .dLion hydrochloride With di-amine 19 (64 mg; 0.10 mmol) in solution in anhydrous methanol (3 ml) is added a 0.098 M hydrochloric methanol solution (2.13 ml;
0.21 mmol). The reaction medium is stirred for 10 min. The expected product is precipitated after addition of ether (20 ml). 50 mg (73%) of the hydrochloride are collected.
Characteristics: Pf = 1 70 ~ C
C31H39N2 ~ l 1Cl ~ M = 649 Solubility test: 2.0 mg in 0.05 ml of water C = 0.06 M
FXF.MPT F 8 General formula I: NRlR2 = a-NH2; X = OH; Y = CH3 4'-Demethyl-4-0 (3-amino-2,3,6-trideoxy ~ -D-ribo-hexopyranosyl) epipodophyllotoxin (compound 26) 25 Stage 1 Tertbutyldimethylsilyl-3- ~ ido-2,3-dideoxy-6-0-tosyl- ~ -D-ribo-hexopyranoside 2 1 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 at stage 1 of example 3 previously cooled to 0 ~ C. After shaking at the same the ~ dlLlre for 1 h, then for 18 h at 20 ~ C, the reaction medium is diluted with CA 02234484 l998-04-08 WO 97/13776 PCT ~ FR96 / 01588 dichloromethane (100 ml). The organic phase is washed with water (2 x 100 ml), dried over MgSO4 and concentrated under reduced pressure. The residue is purified on gel of silica (cyclohexane / EtOAc: 8/2). This provides ~, 0 ~ g of 21 (65%).
Characteristics: TLC: cyclohexane / AcOEt: 2/1; Rf = 0.46 MS: m / z 475 (M + H) +
C 1 gH31N3O6SSi M = 457 St ~ le 2 Tertbutyldimethylsilyl-3-azido-6-iodo-2,3,6-trideoxy- ~ BD-ribo-hexopyranoside 22 Compound 21 (2.02 g; 4.42 rnmol) obtained in stage 1 in solution in 120 ml acetone is heated at reflux for 72 h in the presence of sodium iodide (2.65 g;
17.68 mmol). After cooling, the reaction medium is concentrated under pressure reduced (30 ml) then diluted with dichloromethane (100 ml). After washing with an aqueous 10% sodium thiosulfate solution, then drying over MgSO4 and evaporation under reduced pressure, a residue is obtained which is purified on silica (cyclohexane / EtOAc: 8/2) foll ~ ic ~ nt 1.5 g (82%) of 22.
Characteristics: [a] D20 = - 30 ~ (c = 1.06; CHCl3) MS: m / z 431 (M + NH4) +
C12H24N3O3ISi M = 413 CHN
Calculated 34.87 5.81 10.17 Found 35.07 5.76 10.25 Stage 3 Tertbutyldimethylsilyl-3 -azido-6-iodo-4-O-chloroacetyl-2,3,6-tridesoxy-, BD-ribo-hexopyranoside (compound 23).
Chloroacetyl chloride (396 ml; 5 mmol) is added to a solution of 22 (1.03 g; 2.5 mmol) obtained in the preceding stage in a mixture of dichloromethane (20 ml) and pyridine (404 ~ 11; 5 mmol). After 1 h of stirring at -10 ° C., the medium reaction is diluted with CH2Cl2 (30 ml) and washed with water (3 x 20 ml). Usual treatment, followed by chromatography on silica (cyclohexane / EtOAc: 10/1) 1.1 g (90%) pound 23.

) CA 02234484 1998-04-08 Characteristics: [a] D20 = - 14 ~ (c = 1.03; CHC13) MS: m / z 507 (M + NH4) +
Cl ~ H2sN3O ~ CHSi M = 489 CHN
s Calculated 34.35 5.11 8.59 Found 34.69 5.16 8.22 Stage 4 4'-Demethyl-4-0 (3-azido-6-iodo-2,3,6-trideoxy- ~ -D ribohexopyranosyl) epipodo-phyllotoxin (compound 24) To a mixture of DMEPT 4'-OZ (1 g; 1.85 mmol), 23 (1 g; 2.04 mmol) from the previous stage in anhydrous dichloromethane (100 ml) cooled to -15 ~ C, is added boron trifluoride etherate (BF3.Et20) (455 III; 3.7 mmol). After reaction for 5 h (- 15 ~ C ~ 0 ~ C), the reaction medium is diluted with CH2C12 (100 ml) then, poured into a saturated NaHCO3 solution (200 ml). The organic phase is dried over MgSO4, concentrated under reduced pressure. The crude residue is chromatographed on gel of silica (cyclohexane / AcOEt: 7/3) to provide 24 (0.8 g; 48 ~ / O).
Characteristics: CCM: cyclohexane / AcOEt: 1/1; Rf = 0.58 [a] D20 = - 85 ~ (c = 1.26; CHC13) MS: m / z 909 (M + NH4) +
Pf = 143 ~ C
C37H3sN3O13ClI M = 891 Stage S
4'-Demethyl-4-0 (3-Azido-6-iodo-2,3, 6-tridesoxy-, ~ -D-ribo-hexopyranosyl) 4'-benzyloxy carbonyl epipodophyllotoxin (compound 25) To a solution of azido-glycoside 24 (257 mg; 0.29 mmol) in a CH2Cl2 / MeOH mixture (15 ml, 2/1) is added with OH- resin (AmberliteTM IRA
410). After reaction for 3 h at 20 ~ C, the reaction medium is filtered and then concentrated under reduced pressure to provide 0.22 g of pure (94%).
Characteristics: CCM: cyclohexane / AcOEt: 1/1; Rf = 0.46 [a] D20 = - 57 ~ (c = 1.02; CHCl3) , 0 ~ IFI ~ E

CA 02234484 l998-04-08 WO 97/13776 PCT ~ FR96 / 01588 MS: m / z 833 (M + NH4) +
Pf = 115 ~ C
C3sH34N3O12I M = 815 St ~ 6 4'-Demethyl-4-0 (3-amino-2,3,6-tridesoxy-, BD ribohexopyranosyl) epipodophyllotoxin (compound 26) To a solution of 25 (0.20 g; 0.25 mmol) of the previous stage in 10 ml of ethyl acetate are added triethylamine (100 μl) then palladium-on-carbon has 10% (0.2 g). After stirring for 30 h at ambient temperature in the presence of hydrogen at atmospheric pressure, the catalyst is removed by filtration and the phase organic concentrated under reduced pressure then chromatographed on silica gel (CH2C12 / MeOH: 92/8) to provide 50 mg (38%) of 26.
Characteristics: CCM: CH2C12 / MeOH: 90/10; Rf = 0.33 [a] D20 = - 90 ~ (c = 0.5; CHC13) SM: m / zS30 (M + H) +
Pf = 140 ~ C
C27H31NOlo M = 529 P ~ paldtion of the hydrochloride Amine 26 (70 mg; 0.14 mmol) prec ~ slow in solution in methanol anhydrous (6 ml) is added a 0.098 M hydrochloric methanol solution (1.44 ml;
0.14 mmol). The reaction medium is stirred for 10 minutes. The expected product is precipitated after addition of ether (20 ml). 40 mg (53%) of hydrochloride are collected cry ~ t ~ é
Characteristics: Pf = 164 ~ C
C27EI32NO l oCl M = 565 Solubility test: 2.6 mg in 0.2 ml of water C = 0.02 M
FXFMPT F.9 General formula I: NR1R2 = a-NH2; X = OH; Y - CH2NH2 4'-Demethyl-4-0- (3,6-diamino-2,3,6-trideoxy- ~ -D ribohexopyranosyl) epipodo-phyllotixin (compound 29) Stage 1 4 '-Demethyl-4' -benzyloxycarbonyl-4-0 (3,6-diazido-4-azidoacetyl-2,3,6-tndesoxy-, ~ -D
ribo-hexopyranosyl) epipodophyllotoxin (compound 27) To a solution of 24 (0.45 g; 0.51 mmol) obtained in stage 4 of Example 8 in 10 ml of dimethylformamide is added sodium azide (0.1 g; 1.5 mmol). The reaction 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 MgSO4, concentrated under reduced pressure and gel chromatographed 0 silica (cyclohexane / AcOEt: 7/3) to provide 0.36 g of 27 (90%).
Characteristics: CCM: cyclohexane / AcOEt: 6/4; Rf = 0.44 [a] D20 = - 64 ~ (c = l; CHC13) MS: m / z 831 (M + NH4) +
Pf = 120 ~ C
C37H35NgO 13 M = 813 Stage 2 4 '-Demethyl-4' -benzyloxycarbonyl-4-0 (3,6-diazido-2,3,6-trideoxy-, ~ -D-ribo-hexopyranosyl) epipodophyllotoxin (compound 28) To a solution of azido-glycoside 27 (70 mg; 0.08 mmol) in 3 ml of a CH2Cl2 / MeOH mixture (2/1, v / v) is added to the OH resin: (AmberliteTM IRA 410).
After reaction for 5 h at 20 ~ C, the reaction medium is filtered and then concentrated under reduced pressure to provide 59 mg of pure 28 (94%).
Characteristics: CCM: cyclohexane / AcOEt: 6/4; Rf = 0.25 [a] D20 = - 53 ~ (c = 1.04; CHC13) Pf = 125 ~ C
C3sH34N6ol2I M = 730 Stage 3 4'-Demethyl-4-0 (3,6-diamino-2,3,6-tridesoxy-, ~ -D-ribo-hexopyranosyl) epipodophyllotoxin (compound 29) To a solution of 28 (0.11 g; 0.15 mmol) from the previous stage, in a mixture ~ ~ ',''L' ~ irlE ~

CA 02234484 l998-04-08 - WO 97/13776 PCT ~ FR96 / 01588 ethyl acetate and ethanol (10 ml, 1/1) are added triethylamine (20 ~ 11) then 10% palladium on carbon (70 mg). After stirring for 16 h at ambient temperature in the presence of hydrogen at atmospheric pressure, the catalyst is removed by filtration and the concentrated organic phase under reduced pressure to provide 78 mg 5 (9S%) of 29 ~
Characteristics: TLC: CH2Cl2 / MeOH (NH3): 90/10; Rf = 0.06 C27H32N2Olo M = 544 Preparation of Oichlorlurdrate With di-arnine 29 (78 mg; 0.14 mmol) precederlt ~ in solution in methanol Anhydrous (2 ml) is added a 0.098 M hydrochloric methanol solution (2.92 ml;
0.78 mmol). The reaction medium is stirred for 10 minutes. The expected product is precipitated after addition of ether (20 ml). 10 mg (79%) dihydrochloride are collected crystallized.
Characteristics: Pf = 9 ~~ C
C27H34N2 ~ I oCl2 M = 616 Solubility test: ~, 1 mg in 0.05 ml of water C = 0.07 M

F ~ biological experimentation 20The molecules have been tested in biological experimentation and have shown their interest as an anticancer agent in tests of P 388 leukemia in vivo in mouse. This test is commonly used in the field of substance research anticancer drugs (Protocols for screening chemical agents and natural products against animal tumors and other biological systems, R. Geran, NH Greenberg, MM
25 MacDonald, AM Sch-lm ~ h ~ r and BJ Abbott, Cancer Chemotherapy reports 1972, 3, N ~ 2).
However, this experimental model is extremely chemosensitive and very many compounds show good activity which makes this test not very discriminating We have modified the test protocol to make it more selective.
30 ~ ~ -lmini ~ tration of tumor cells is done intravenously and not by CA 02234484 l998-04-08 intraperitoneally. They are thus disseminated by circulation throughout the body quickly. The aminictration of the product to be tested is then made by intraperitoneally. Two parameters are defined to highlight the activity of compounds:
5 ~ determination of the effective dose 50 (DEso) which represents the single minimum dose of the compound to be mined to obtain a significant survival of the nim by compared to ~ nim ~ lly untreated controls, ~ determination ~ the maximum survival time of ~ nim ~ lx whatever the dose ~ lminictrée by single injection. Being able to minify a large dose of the compound and observe a significant survival, makes it possible to obtain a measurement the maximum therapeutic efficacy of the product that can be achieved.

Tumor oroine P 388 leukemia was chemically induced in 1955 by 3-methylcholanthrene on a DBA / 2 mouse (Am. J. Pathol. 33, 603, 1957).

Colorless procedure Tumors are m ~ int ~? N ~ les by weekly passages in the form of ascites in the peritoneum of DBA / 2 mice (original line) and the experiments are performed on CDFl hybrid female mice (bal b / c female XDBA / 2 males) of 20 + 2 g (Cancer chemother, Rep. 3, 9, 1972). Tumor cells are implanted intravenously (106 cells per mouse) on day 0. The ~ nim ~ l ~ x are randomized and divided into groups of 2 for each series.
The sllbst ~ nces ~ ntitllmorale5 are ~ -lminictrées intraperitoneally (ip) a 2 ~ day after inoculation of leukemia cells (acute treatment). The solutions are injected at the rate of 10 ml / lcg of mice. The criterion for evaluating anti-tumor activity is the prolongation of the survival of treated Zlnim ~ llx. 86% of mice die on the 7th day after tumor transplant, A substance will be considered active if it induces survival greater than 8 days.
The following table makes it possible to highlight the aqueous solubility of - WO 97/13776 PCT ~ FR96 / 01588 products of the invention, e ~; awarded in mg / ml, the activity of these compounds in terms of DEso, survival expressed in days or in T / C%, which represents the relationship between survival mean of the group of ~ nim ~ n ~ treated and mean survival of the group of ~ nim ~ n ~ c control.
Solubility P 388 Survival aqueusemg / ml ED50 (mg / kg) ma ~; imum (J) T / C%
Animals control with 6-8 a-lmini ~ tration iv of (median-7) tumor cells Compounds Etoposide 0.01 10 19 271 Teniposide <0.01 20 15 214 1 1 25 1 0 1 7 2 ~

7 10 14 ~ 00 It thus appears that the compounds of the invention have retained the level of activity benchmark compounds like Etoposide and have the added benefit of having aqueous solubility advantageous for formulation and administration.

Claims (11)

1) A compound of general formula I

in which the group at 3" N(R1 R2) is in the .beta position. (2-desoxy D series Arabino) or a (2-desoxy D ribo series) with respect to the ring, R1 and R2 identical or different represent a hydrogen atom, an alkyl group from C1 to C6, which can form a cycle, this cycle possibly comprising a heteroatom such as an oxygen or a nitrogen, a C1 to C6 aminoalkyl or cyanomethyl group, X and Y can be the same or different and represent OH, CH3, CH2-NH2, X
and Y can also be linked and constitute a cycle, for example, a 2-methyl 1,3 dioxane, thus forming a 4,6-ethylidene-like bicyclic osidic skeleton 3 amino 2,3 dideoxy glycoside, and its addition salts with acids mineral or organic, pharmaceutically acceptable. 2) A compound of general formula I according to claim 1, characterized in that that the NR1R2 group is an NH2 or N(CH3)2 group. 3) A compound of general formula I according to claim 1, characterized in that that the NR1R2 group represents an amino group substituted, once or twice by a methyl, CH2CN or a CH2-CH2-NH2. 4) A compound of general formula I according to one of claims 1 to 3, characterized in that X and Y form a cycle with an OCH(CH3)OCH2 sequence. 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:
~ 4'-Demethyl-4-0(3-amino-4,6-ethylidene-2,3-dideoxy-.beta.-D-arabinohexopyranosyl) epipodophyllotoxin, ~ 4'-Demethyl-4-0(3-amino-4,6-ethylidene-2,3-dideoxy-.beta.-D-ribohexopyranosyl) epipodophyllotoxin, ~ 4'-Demethyl-4-0(3-dimethylamino-4,6-ethylidene-2,3-dideoxy-.beta.-D-arabino-hexopyranosyl) epipodophyllotoxin, ~ 4'-Demethyl-4-0(3-dimethylamino-4,6-ethylidene-2,3-dideoxy-.beta.-D-ribo-hexopyranosyl) epipodophyllotoxin, ~ 4'-Demethyl-4-0(3-cyanomethylamino-4,6-ethylidene-2,3-dideoxy-.beta.-D-ribo-hexopyranosyl) epipodophyllotoxin, ~ 4'-Demethyl-4-0(3-(N-morpholino)-4,6-ethylidene-2,3-dideoxy-.beta.-D-ribo-hexopyranosyl) epipodophyllotoxin, ~ 4'-Demethyl-4-0[3(2-aminoethylamino)-4,6-ethylidene-2,3-dideoxy-.beta.-D-ribo-hexopyranosyl)] epipodophyllotoxin, ~ 4'-Demethyl-4-0(3-amino-2,3,6-trideoxy-.beta.-D-ribohexopyranosyl) epipodophyllotoxin, ~ 4'-Demethyl-4-0(3,6-diamino-2,3,6-trideoxy-.beta.-D-ribohexopyranosyl) epipodophyllotoxin. 6) Compounds according to claim 4, characterized in that they are in form of hydrochlorides. 7) Process for the preparation of a compound of general formula I according to the claims 1 to 4, characterized in that a compound of formula III or IV or V

with 4'-Demethyl 4'-benzyloxycarbonyl epipodophyllotoxin with BF3 etherate, or trimethyl silyl trifluoromethanesulforate in an inert solvent at low temperature;
in formula III and IV, the substituent at position 3 may be .alpha. or .beta., NR1R2 can be an amino protected by a Z group, in formula V, P represents an alcohol protecting group and the products resulting from this condensation are deprotected and hydrogenated to provide the compounds of formula I, the primary amines in position 3 of the glycosyl are methylated by formalin and sodium cyanoborohydride. 8) Process for preparing a compound of formula I according to claim 7 characterized in that the compound of formula IV is prepared by reacting a mixture diacetoxy azido glycoside VI

with tert-butyl dimethylsilyl chloride in the presence of imidazole, in that one separates the products resulting from this reaction, in that each of these products are deacetylated, cyclized to 4,6-ethylidene with acetaldehyde acetal in acid medium catalytic. 9) As a medicament the compounds of formula I according to claims 1 to 6. 10) Pharmaceutical compositions characterized in that they contain at least one compound of formula I according to one of claims 1 to 6 and an excipient appropriate. 11) Use of a compound of formula I, according to one of claims 1 to 4, for the preparation of a medicament intended for the treatment of cancers and, in particular forms of cancer, such as small cell lung cancer, testicular cancer, embryonic tumors, neuroblastomas, kidney cancer, placental choriocarcinomas, mammary adenocarcinomas, cancers colorectal, melanoma, Hodgkin's and non-Hodgkin's lymphomas and acute leukemia.