FR2630915A1 - Novel glycals, anthracyclines obtained using these glycals and use of these anthracyclines as medicaments - Google Patents

Abstract

The invention relates to novel glycals, to anthracyclines obtained using these glycals and to the use of these anthracyclines as medicaments. The glycals are obtained from L-fucose and/or its analogues of formula 1': and correspond to the formula 2 below: in which: R represents an optionally substituted linear or branched alkyl group or an optionally substituted aryl group, with the exception of a hydrogen atom, a CH2OH group or a group CH2OR3, R3 having the same meaning as R.

Description

 The present invention relates to novel glycals, to anthracyclines obtained using these glycals and to the use of said anthracyclines as medicaments.

 Anthracyclines are an important family of anti-cancer agents. Their major disadvantage is their cardiotoxicity; cardiac involvement is a function of the total dose administered; however, this may vary depending on the anthracycline.

L-Fucose is of great biological interest since it is one of the constituents of several blood group antigens and several glycosides, derived from deoxy-2-L-Fucose, easily obtained from the
L-Fucose, exhibit antitumor and antibiotic activities in anthracycline series.

 In particular, the glycoside of Formula 3a (D.

présente une activité antitumorale elevée in vive, simi- laire à celle de la daunorubicine de formule 4a (D. HORTON et al., J. Med. Chem., 1976,22,406).

HORTON et al., Carbohydr. Res., 1977, 57, C36)

has a high anti-tumor activity in vivo, similar to that of daunorubicin of formula 4a (D. HORTON et al., J. Med Chem, 1976, 22, 406).

qui présentent également une activité antitumorale remarquable.Deoxy-2-L-Fucose is also the central component of many anthracycline-series trisaccharides, such as aclacinomycin A of Formula 5

which also exhibit remarkable antitumor activity.

 Numerous efforts are continuing in this field, with the aim of improving the therapeutic index of anthracyclines, but also to find compounds active on anthracycline-resistant cancer cell strains currently used clinically. One of the most promising modifications seems to be that of the glycosidic part which is involved in all the steps of the mode of action of these compounds (transport, intracellular accumulations, complexation with the DNA ...).

That is why, the Applicant, pursuing the study undertaken concerning the synthesis of new glycosides of high therapeutic value (cf.
French Patent Nos. 84 03634 and 85 10063), has developed a new and economically interesting method of synthesizing L-Fucose and analogues thereof, which are indispensable for the preparation of new anthracyclines. .

est un sucre obtenu difficilement par extraction d'espèces de Fucus variées et est donc d'un prix de revient élevé.Unfortunately, L-Fucose (or deoxy-6-L galactose) of Formula 1

is a sugar obtained with difficulty by extraction of various species of Fucus and is therefore a high cost.

Several total syntheses have already been proposed, but.

they are characterized by a high number of steps or by the need for a separation step which is poorly adapted to the preparation of large quantities (DEFAYE et al., Carbohydr Res., 126, 165, 1984).

 It has therefore been the object of the present invention to provide novel anthracyclines which better meet the requirements of the practice than the anthracyclines of the prior art, their process of preparation as well as the sugars and glycals used for their preparation. preparation.

dans laquelle
R représente un groupe alkyle éventuellement substitué, li neaire ou ramifié ou un groupe aryle éventuellement substitué, à l'exception d'un atome d'hydrogène, d'un groupe me- thyle, d'un groupe CH2OH ou d'un groupe CH2OR3, R3 ayant la même signification que R
Ri et R2 sont toujours différents et représentent un atome d'hydrogène ou un groupe hydroxyle, étant donné que lorsque R1 est un atome d'hydrogène et R2 est un groupe OH, on obtient un stéréo isomère e et lorsque Ri est un groupe hydroxyle et R2 est un atome d'hydrogène, on obtient un stéréoisomère a sont décrits dans la demande principale n' 88 05969 du 4 mai 1988.Analogs of L-Fucose, corresponding to the general formula 1 '

in which
R represents an optionally substituted, branched or branched alkyl group or an optionally substituted aryl group, with the exception of a hydrogen atom, a methyl group, a CH 2 OH group or a CH 2 OR 3 group; , R3 having the same meaning as R
R 1 and R 2 are always different and represent a hydrogen atom or a hydroxyl group, since when R 1 is a hydrogen atom and R 2 is an OH group, a stereo isomer e is obtained and when R 1 is a hydroxyl group and R2 is a hydrogen atom, a stereoisomer a is described in the main application No. 88 05969 of May 4, 1988.

dans laquelle
R représente un groupe alkyle éventuellement substitué, linéaire ou ramifié ou un groupe aryle éventuellement substitué, à l'exception d'un groupe CH2OH, d'un groupe
CH20R3, R3 ayant la même signification que R.The subject of the present invention is glycals obtained from L-Fucose and / or its analogues, from
Formula 2

in which
R represents an optionally substituted, linear or branched alkyl group or an optionally substituted aryl group, with the exception of a CH 2 OH group, of a group
CH20R3, R3 having the same meaning as R.

dans laquelle R a la même signification que précédemment
Ri est un hydrogène et R2 un groupe acétoxy
l'isomère 18 ss peut être séparé par précipitation sélective.Glycals of Formula 2 are obtained in a known manner according to the method of B. Iselin and T. Reichstein (Helv Chim Acta, 1944, 27, 1200) which comprises the following steps
First step: peracetylation of a mixture of anomers a and ss of Formula 1 'to obtain a derivative in the form of isomers a and 13 of Formula 18

in which R has the same meaning as above
Ri is a hydrogen and R2 is an acetoxy group
the 18S isomer can be separated by selective precipitation.

Second step treatment of the mixture 18 a and
B with hydrobromic acid, to obtain an intermediate brominated derivative which, subjected to the action of zinc provides a glycal of Formula 2, according to the following scheme III

SCHEMA III

The subject of the present invention is, moreover, a process for the preparation of novel anthracyclines from a glycal according to the invention, characterized in that
at. said glycal of Formula 2 is converted into a halogenated derivative by adding a compound of formula HX anhydrous, wherein X is a halogen, to obtain a compound of Formula 19, according to scheme IV.

SCHEMA IV

b. then said compound of Formula 19 is coupled to a suitable anthracyclinone, by a reaction of the type
Koenigs-Knorr.

dans laquelle
R est tel que défini ci-dessus
Ri représente un groupe acétyle ou un atome d'hydrogène.According to one embodiment of said method, said compound 19 is coupled with daunomycinone and provides a product of Formula 20 '

in which
R is as defined above
R 1 represents an acetyl group or a hydrogen atom.

According to another embodiment of said method, said compound 19 is coupled. with ss rhodomycinone and provides a product of Formula 21 '.

dans lequel R et Ri ont les mêmes significations que précédemment.in which,
R is as defined above
R 1 represents an acetyl group or a hydrogen atom R 4 represents a hydrogen atom or the radical of Formula 19 ',

wherein R and R 1 have the same meanings as before.

 The glycosides obtained are purified by chromatography.

 In addition to the foregoing, the invention also comprises other provisions which will emerge from the description which follows.

 The invention relates more particularly to the novel analogues of L-Fucose and new glycals obtained from said analogs as well as new anthracy turkeys, and their methods of preparation, as well as compositions, in particular therapeutic compositions, in which said derivatives, for their use in anti-cancer chemotherapy.

 These new anthracyclines have remarkable cytotoxic and antitumor properties, as specified in the pharmacological tests below.

 Among the various anthracyclines according to the present invention, having very interesting pharmacological results, mention should be made in particular of Di-O- (di-O-acetyl-3 ', 4' dideoxy-2 ', 6' -aL -threo-hexopyranosyl) -7,10 ss-Rhodomycinone of Formula 22a O- (di-O-acetyl-3 ', 4'-dideoxy-2', 6'-α-threohexopy-rannosyl) -7 ss-Rhodomycinone of Formula 21a Di-O- (di-O-acetyl-3 ', 4'-trideoxy-2', 6 ', 7'-α-threoheptopyranosyl) -7,10 B-Rhodomycinone of Formula 22b O- (di-O-acetyl-3 ', 4'-trideoxy-2', 6 ', 7'-α-threohep-thyranosyl) -7-Rhodomycinone of Formula 21b di-O- (di-O- 3'-acetyl-4 'hexadesoxy-2', 6 ', 7', 8 ', 9', 10'-α-threo-decapyranosyl) -7,10 ss-Rhodomycinone of Formula 22, O- (di-O) 3'-acetyl-4'-hexadoxoxy-2 ', 6', 7 ', 8', 9 ', 10' -α-L-threo decapyranosyl) -7-Rhodomycinone of Formula 21c. 3'-acetoxy-4'-acetyl-4'-desamino-3'-methyl-6'-daunorubicin of Formula 20b. 3'-acetoxy-3'-O-acetyl-3'-desamino-6'-propyl-6 'daunorubicin of Formula 20c.

 The invention will be better understood using the additional description which follows, which refers to examples of implementation of the method, object of the present invention as well as examples highlighting the therapeutic properties of said glycosides.

 In I.R., the bands are given in cm-1.

 In NMR, the chemical shifts are given in ppm relative to the tetramethylsilane (TMS) used as reference.

EXAMPLE 1 SYNTHESIS OF GLYCALS, WITH R = C2H5
First stage
6-Methyl Fucose (5g, 0.028 mol) is suspended in pyridine (50ml) and acetic anhydride (25ml). The reaction mixture is stirred for 48 hours at room temperature. The resulting solution is thrown on ice water and then extracted in the usual way with
CH2Cl2. 4.95 g (Yield: 51%) of a mixture of tetraacetates 18b and 18b are obtained.

Oil (a) D = -32.4 (c = 1.11, CHCl3)
IR (CH 2 Cl 2): 2990, 1750, 1370, 1220, 1090, 1060, 900.

NMR (CDCl3). 0.92 (t, J = 7Hz, 1H); 1.5 - 1.8 (m, 2H) 1.99 (s, 3H); 2.04 (s, 3H); 2.12 (s, 3H); 2.18 (s, 3H) 3.65 (t, J = 6.5Hz, 1H); 5.06 (dd, J1 = 3.5Hz, J2 = 10Hz, 1H); 5.2 - 5.4 (m, 2H); 5.66 (d, J = 8Hz, 1H).

Second step
A solution of diacetates 18b and ss (* 5g) in acetic acid (1.25ml) and acetic anhydride (1.25ml) is stirred at 0 ° C. 30% HBr solution in acetic acid (9 ml) and stirred until the temperature has returned to 20 ° C.

 This solution is then added slowly to a solution cooled to -10 C of sodium acetate (12.5 g) in acetic acid-water 50/50 v / v (30 ml) to which has been added (at this point). temperature) a solution of CuSO 4, 5H 2 O (0.9 g) in 2.75 ml of water and pulverized zinc (9 g). After the addition is complete, stirring is continued for two hours at -10.degree.

 It is then filtered on celite, washed twice with a solution of CH 3 COOH-H 2 O, then extracted with CH 2 Cl 2. After washing with a saturated solution of bicarbonate, then with water and finally drying over Na 2 SO 4, an oil is obtained which is purified by fast filtration on silica gel (eluent ether-petroleum ether 70/30).

The glycal of Formula 2b is thus obtained (2.43 g, Yield: 73%)
F: 47-48 C ta) D = + 3.7 (c = 1.06, CHOCl 3)
IR (CH 2 Cl 2): 3000, 2900, -1745, 1655, 1375, 1240, 1155,
1100, 1080, 1035, 910.

NMR (CDCl 3): 0.97 (t 1, J = 7 Hz, 3H); 2.01 (s, 3H); 3.93
(t., JF 7 Hz, 1H); 4.65 (dt, J1 = 6 Hz,
Jz = 2 Hz, 1H); 5.37 (d, J = 4 Hz, 1H)
5.56 (bs, 1H); 6.46 (dd, J1 = 6 Hz,
J2 = 2 Hz, 1H).

EXAMPLE 2: GLYCAL SYNTHESIS IN BUTYL SERIES, WITH
R = C4H9
First stage
The acetylation reaction is carried out under the same conditions as in Example 4. A mixture of tetraacetates 18c a and 18c ss is thus isolated in the form of an oil.

 -This mixture can be recrystallized partially in ether tetraacetate B of Formula 18c.

F: 111 C (a) D = -31 '(c = 1, CHCl3)
IR: 3070, 2980, 2890, 1750, 1370, 1220, 1095, 1065,
1050.

NMR (CDCl 3): 0.86 (t., J = 7 Hz, 3H); 1.29 (m, 4H); 1.99
(s, 3H); 2.04 (s, 3H); 2.11 (s, 3H); 2.17
(s, 3H); 3.74 (dd, J1 = 8 Hz, J2 = 3.5 Hz,
1H); 5.05 (dd, J1 = 3.5 Hz, J2 = 10 Hz,
1H); 5.29 (d, J = 9Hz, 1H); 5.34 (d., J = 8
Hz, 1H); 5.47 (d, J = 9 Hz, 1H).

Second step
The reaction is carried out as in Example 4. The glycal of Formula 2c (Yield: 60%) is obtained.

Oil (a) D = -24 ° (c = 1.68, CHCl3)
IR (CH 2 Cl 2): 2970, 2950, 2845, 1740, 1375, 1230, 1150,
1120, 1050.

NMR (CDCl 3): 0.98 (t 1, J = 7 Hz, 3H); 1.34 (m); 2.01 (s,
3H); 3.99 (dd, Ji = 8 Hz, J2 = 4 Hz, 1H)
4.63, (dt, J1 = 6 Hz, J2 = 2 Hz, 1H); 5.33
(d., J = 4 Hz, 1H); 5.56 (bs, 1H)
6.47 (dd, J1 = 8 Hz, J2 = 2 Hz, 1H).

EXAMPLE 3 GLYCOSIDATION OF BETA-RHODOMYCINONE WITH A
RADICAL FORMULA 19 'IN WHICH R IS A GROUP CH3
To a solution of ss-Rhodomycinone (300 mg, 0.78 mmol) in anhydrous methylene chloride (150 ml) is added successively HgO (1.014 g), HgBr 2 (297 mg) and then molecular sieves 3 A (3 g). .

 This suspension is stirred under nitrogen at ambient temperature during the addition of a solution of 1-chloro-di-O-acetyl-3,4-trideoxy-1,2,6-L-galactose (19 a) (488 mg). in CH2Cl2 (15 mL).

 After 12 hours at room temperature, the reaction mixture is filtered (the insoluble fraction is washed with CH 2 Cl 2) and then extracted.

 The organic phase is washed successively with a solution of bicarbonate and then with water. After drying, an oil is obtained which is chromatographed on silica gel (eluent: CH2Cl2-MeOH-AcOH: 99-1-0.2 v / v).

 The bisglycoside is thus successively isolated at the 7- and 10-positions (10-15%) and then at the 7-position glycoside (40-50%).

 Di-O-Di-O- (di-O-acetyl-3 ', 4'-dideoxy-2', 6 '- α-L-threohexo-pyranosyl) -7,1-rHodomycinone of Formula 22a.

F: 157-158-C (a) D = + 399 (CHCl3)
IR (CHCl3). 3680, 3340, 2980, 2940, 1735, 1600, 1570,
1435, 1395, 1360, 1275, 1245, 1230, 1195,
1160, 1110, 1075, 1010, 955.

NMR (CDCl 3): 1.11 (t, J = 7 Hz, 3H); 1.92 (s, 6H); 2.15
(s, 3H); 2.18 (s, 3H); 3.32 (s, 1H, OH)
4.16 (s, J = 6.5 Hz, 1H); 4.29 (q, J = 6.5
Hz, 1H); 5.01 (s, 1H); 5.0-5.3 (m complex,
4H); 5.53 (d, J = 2.5 Hz, 2H); 7.32 (d.,
J = 8 Hz, 1H); 7.72 (t., J = 8 Hz, 1H)
7.89 (d, J = 8 Hz, 1H); 12.12 (s, 1H, OH)
12.87 (s, 1H, OH); 13.72 (s, 1H, OH).

O- (di-O-acetyl-31,4 'dideoxy-2', 6 '- α-L-threohexopyanyanosyl) -7 B-Rhodomycinone of Formula 21a
F: 136-140 C (α) D = + 174 (CHCl3)
IR (CH 2 Cl 2): 3700, 3600, 2990, 2940, 1735, 1600, 1575,
1415, 1360, 1230, 1200, 1160, 1075, 1035,
1015, 90, 960.

NMR (CDCl3): 1.13 (t., J = 7 Hz, 3H); 1.23 (d., J = 6 Hz,
1H); 1.92 (s, 3H); 2.18 (s, 3H); 3.67 (s,
1H); 4.35 (q, J = 6Hz, 1H) 4.89 (s, 1H)
5.10 (br s, 1H); 5.21 (s, 1H); 5.54 (s,
1H); 7.30 (d, J = 8 Hz, 1H); 7.69 (t, J =
8 Hz, 1H); 7.86 (d, J = 8h, 1H); 12.09
(s, 1H); 12.89 (s, 1H); 13.55 (s, 1H).

EXAMPLE 4 GLYCOSIDATION OF ss-RHODOMYCINOKE, WITH A
RADICAL OF YORUBLE 19 ', IN WHICH R IS A GROUP
ETHYLE:
The reaction is carried out as in Example 6, using chloro-1-di-O-acetyl-3,4-methyl-6-tridoxy-1,2,6-L-galactose 19b.

 There is thus obtained Di-O- (di-O-acetyl-3 ', 4'-trideoxy-2', 6 ', 7' - α-L-threoheptopyranosyl) -7,10 B-Rhodomycinone of Formula 22b.

F 196 - 199 (dec.) (A) D = + 464 '(CHCl3, c = 1.66)
IR (CHCl 2): 3700, 3540, 2990, 2940, 1735, 1600, 1575,
1415, 1365, 1270, 1240, 1200, 1160, 1115,
1060, 1015, 985, 970.

NMR (CDCl3): 1.93 (s, 6H); 2.14 (s, 3H); 2.18 (s, 3H)
3.31 (s, 1H, OH); 3.89 (t., J = 7 Hz, 1H)
4.01 (t 1, J = 7 Hz, 1H); 4.99 is, 1H); 5
5.25 (m complex, 4H); 5.29 (s, 1H); 5.50
(d, J = 2.5 Hz, 1H); 5.58 (d, J = 2.5 Hz,
1H); 7.32 (d, J = 8Hz, 1H); 7.72 (t., J =
8Hz, 1H); 7.89 (d, J = 8 Hz, 1H); 12.09
(s, 1H, OH); 12.88 (s, 1H, OH); 13.69 (s,
1H, OH).

O- (di-O-acetyl-3 ', 4'-trideoxy-2', 6 ', 7' -α-L-threo-heptopyranosyl) -s-rhodomycinone of Formula 21b
F 133-135 ° C (dec) (a) D = + 208 ° C (c = 1.96, CHCl 3)
IR (CH 2 Cl 2): 3700, 3540, 3000, 2950, 1715, 1610, 1580,
1420, 1360, 1250, 1220, 1170, 1025.

NMR (CDCl 3): 1.03 (t 1, J = 7 Hz, 3H); 1.13 (t, J = 7 Hz,
3H); 1.93 (s, 3H); 2.18 (s, 3H); 3.15 (s,
broad, 1H, OH); 3.76 (s, 1H, OH); 4.10 (t.,
J = 7 Hz, 1H); 4.85 (s, 1H); 5.07 (d., J =
2 Hz, 1H); 5.32 (s, 1H); 7.63 (t, J =
8 Hz, 1H); 5.58 (d, J = 2 Hz, 1H); 7.20
(d, J = 8 Hz, 1H); 7.73 (d, J = 8 Hz,
-1H); 11.94 (s, 1H, OH); 12.75 (s, 1H, OH)
13.40 (s, 1H, OH).

EXAMPLE 5 GLYCOSIDATION OF B-RHODONYCINONE, WITH A
RADICAL FORMULA 19 ', IN WHICH R IS A BUTYL GROUP
The reaction is carried out as in Example 6, using 1-chloro-di-O-3,4-acFtyl-6-propyl-1,2,6-trideoxy-L-galactose 19c.

There is thus obtained di-O- (di-O-acetyl-3 ', 4' hexadésoxy-2 ', 6', 7 ', 8', 9 ', 10'-α-threo-decapyranosyl) -7,10 ss-Rhodomycinone of Formula 22c
F: 190 - 192tC (change of state: 163-167 ° C) (a) D = + 320 '
IR (CH2Cl2): 3710, 3580, 2980, 2960, 2890, 1740, 1605,
1585, 1465, 1410, 1375, 1245, 1205, 1170,
1130, 1070, 1025, 1000, 980.

NMR (CDCl 3): 1.94 (s, 6H); 2.17 (s, 6H); 3.31 (s, 1H)
3.9-4.2 (m, 2H); 5.01 (s, 1H); 5.0-5.45 (m,
5H); 5.50 (brs, 1H); 5.60 (s wide,
1H); 7.32 (d, J = 8 Hz, 1H); 7.72 (t., J =
8 Hz, 1H); 7.92 (d, J = 8 Hz, 1H); 12,13
(s, 1H); 12.91 (s, 1H); 13.70 (s, 1H).

O- (di-O-acetyl-3 ', 4'hexadesoxy-2', 6 ', 7', 8 ', 9'10' - α -L-threo decapyranosyl) -7 B-Rhodomycinone of Formula 21c
F: 110-112-C (a) n: + 179 (CHCla)
IR (CH 2 Cl 2): 3580, 3530, 2960, 2920, 2860, 1735, 1600,
1575, 1455, 1410, 1360, 1240, 1190, 1160,
1130, 1115, 1065, 12020, 990, 965.

NMR (CDCl 3): 0.94 (t., J = 6Hz, 3H); 1.13 (t., - J = 7 Hz,
3H); 1.94 (s, 3H); 2.17 (s, 3H); 3.73 (s,
1H, OH); 4.14 (brs, 1H); 4.90 (s, 1H)
5.13 (brs, 1H); 5.29 (s, 1H); 5.59 (d.,
J = 2 Hz, 1H); 7.32 (d 1, J = 8 Hz, 1H)
7.69 (t 1, J = 8 Hz, 1H); 7.84 (d, J = 8 Hz,
1H); 12.09 (9, 1H, OH); 12.84 (s, 1H, OH)
13.55 (s, 1H, OH).

EXAMPLE 6 GLYCOSIDATION OF DAUNOMYCINONE
It is carried out according to the same procedure as that used for α-Rhodomycinone (Example 5 to 8).

- ethyl series
The 3'-acetoxy-3'-acetyl-4'-desamino-3'-methyl-6'daunorubicin of Formula 20b is obtained.

F: 126-131 ° C (a) b = 216 ° (c = 0.27, CHCl 3)
IR (CH 2 Cl 2): 3500, 1730, 1710, 1620, 1580, 1425, 1380.

NMR (CDCl3): 0.99 (t, 3H); 1.95 (s, 3H); 2.16 (s, 3H)
2.43 (s, 3H); 2.92 (d, J = 19 Hz, 1H)
3.25 (d, J = 19 Hz, 1H); 3.98 (t., 1H)
4.10 (s, 3H); 4.40 (s, 1H); 5.05 (dm,
1H); 5.30 (bs, 2H); 5.63 (d., J =
3 Hz, 1H); 7.40 (d, 1H); 7.79 (t, 1H)
8.05 (d, 1H); 12.53 (s, 1H); 13.27 (s,
1H).

- Butyl series
Acetoxy-3'-O-acetyl-4'-deamino-3'-propyl-6 'daunorubicin of Formula 20c is obtained.

F: not measurable (no defined melting point).

(a) D = 226 '(c = 0.2, CHCl3)
IR (CH 2 Cl 2): 3500, 2960, 2940, 1720, 1705, 1605, 1425,
1375.

NMR (CDCl3): 0.91 (t, 3H); 1.94 (3H); 2.17 (s, 3H)
2.43 (s, 3H); 2.84, (d, J = 19 Hz); 3.19
(d, J = 19 Hz); 4.09 (OCH3, 3H); 4.39 (s
large) ; 5.06 (dm, 1H); 5.29 (s large,
2H); 5.64 (broad d, J = 3.2 Hz, 1H); 7.40
(d., 1H); 7.78 (t, 1H); 8.01 (d., 1H)
11.60 (s, 1H); 12.36 (s, 1H).

PHARMACOLOGICAL REPORT ON GLYCOSIDES
ACCORDING TO THE INVENTION.

I - PROLIFERATION TEST
A - Operating protocol (MTT reduction)
L1210 A549 and HT29 tumor cells, at a concentration of 5 × 10 3 / ml in RPMI medium, are incubated in 96-well microtiter plates for 72 hours (37 ° C., 5% CO2, 95% humidity). relative) with different concentrations of each of the glycosides (anthracyclines) according to the invention.

 The controls consist of tumor cells exposed to a culture medium. Four wells are prepared for each glycoside concentration and for the control.

After 65 hours, 50 μl of MTT (2.5 mg / ml in PBS) are added.

 MTT will be reduced in the presence of living cells to an insoluble red formazan dye. After another 7 to 24 hours of incubation, the supernatant is removed. The formazan dye is solubilized by the addition of 100 μl of DMSO in each well, followed by gentle stirring.

 The extinction is measured for each well at 492 nm (Multiscan 340 CC Fa Flow meter).

B - Results
The results are expressed as the ratio of extinction after incubation with the glycosides to the extinction achieved with the controls. The coefficient of variation is less than 15%.

 The results are shown in Table r below.

The products tested are as follows
TABLE I

<tb><SEP> Cisc <SEP> (g / ml) <SEP>
<tb><SEP> TEST <SEP> AT <SEP> MTT
<tb><SEP> L1210 <SEP> HT <SEP> 29 <SEP> A <SEP> 549
<tb> 21 <SEP> b <SEP> 0.51 <SEP>><SEP> 1 <SEP> 0.46
<tb> 22 <SEP> b <SEP>><SEP> 1 <SEP>><SEP> 1 <SEP>><SEP> 1 <SEP>
<tb> 21 <SEP> c <SEP>><SEP> 1 <SEP>><SEP> 1 <SEP>><SEP> 1
<tb> 22 <SEP> c <SEP>><SEP> 1 <SEP>><SEP> 1 <SEP>><SEP> 1
<tb> 20 <SEP> b <SEP>> 1 /> 10 <SEP>'0.35/> 10 <SEP>> 1 /> 10
<tb> 20 <SEP> c <SEP>><SEP> 1 <SEP>><SEP> 1 <SEP>><SEP> 1
<tb> 21 <SEP> a <SEP> 0.03 <SEP> 0.038 <SEP> 0.086
<tb> II - EFFECT ON L1210 LEUKEMIC CELLS
A - Operating protocol
The test is carried out according to the procedure of
Hamburger and Salmon, with the few modifications specified below.

The medium used is replaced by a medium of
McCoy 5A. The number of cells placed in the dishes is reduced to 5 × 10 cells / case because of the rapid growth of L 1210 leukemia cells.

 The cells are incubated for 1 hour at 37 ° C. in the presence of different concentrations of the test substance. The cells are then washed twice with McCoy 5A and plated on agarose plate according to the Hamburger and Salmon method.

 Moreover, the test is carried out in parallel with continuous incubation with different concentrations of the substance studied by introducing said substance into the agarose layer before the cells are cultured.

 The boots are placed in an oven at 37 ° C. under an atmosphere of 5% CO 2, 20% O 2 and a relative humidity of 95%, for 5 to 7 days. After this period, colonies greater than 60 μm in diameter are counted using an inverted microscope.

B - Results
The results summarized in Table II, below are expressed as percentage of colonies formed from treated L 1210 cells compared to untreated controls. The coefficient of variation, when the experiments are repeated, is less than 15%.

TABLE II

<tb><SEP> TEST <SEP> ON <SEP> CELLS <SEP> LEUKEMICS
<tb><SEP> experience <SEP> of a <SEP> hour
<tb><SEP> experience <SEP> in <SEP> continuous <SEP> experience <SEP> of a <SEP> hour
<tb><SEP> L1210 <SEQ> HT <SEP> 29 <SEP> A <SEP> 549 <SEQ> L121G <SEP> HT <SEP> 29 <SEP> A <SEP> 549
<tb> 21 <SEP> b <SEP> 0.8
<tb> 21 <SEP> a <SEP> 0.057 <SEP> 0.38
<tb> 22 <SEP> a <SEP> 1,2 <SEP>><SEP> 1
<Tb>
III - STUDY OF THE EFFECT OF COMPOUND 21a, IN VIVE:
A - Operative protocol:
- Preparation of L1210 cells
Ascites fluid was removed under aseptic conditions in DBA2 mice (female, 18-20 g) on the seventh day after implantation.

 The ascites is washed three times with PBS, counted and finally diluted in PBS, to obtain 106 cells / 0.2 ml.

- Transfer of cells for the maintenance of the cell line
106 cells in 0.2 ml of PBS are injected into the peritoneum of DBA2 mice for cell line propagation.

 The transfer is done once a week.

- Transfer of cell lines for testing
106 cells in 0.2 ml of PBS are injected intraperitoneally into BDF1 mice (female 18-20 g), 6 animals / group are used for each substance concentration and for control.

B - Evaluation of the effect
a) the animals are weighed on the first day and the fifth day after the administration of the drug. A weight loss of more than 20% on the fifth day is used as an indicator of the toxic effect of the product.

 (b) at the end of the experiment (death of all animals or sixtieth day of the test), the average survival time of the animals, in all groups with more than 65% of survivors on the fifth day, is assessed according to standard procedures.

Surviving animals on the sixtieth day are considered long-term survivors (LTS: Long
Time Survivors) and are listed separately.

From the average survival time of the treated groups (MUST) and the control groups (MSTc), the antitumor effect (T / C) is evaluated according to the following formula MSTT
T / C% = ----- x 100
CSTM
T / C values of more than 125% are considered indicative of significant antitumor effect.

The antitumor effect is specified in the
Table III, below
TABLE III

<tb><SEP> L1210
<tb> name <SEP> program <SEP> dose <SEP> T / C <SEP> LTS * <SEP> Deaths
<tb><SEP> of adminis- <SEP> optimal <SEP> 8 <SEP><SEP> toxic
<tb><SEP> treatment <SEP> (mg / kg / Inj.)
<tb><SEP> 21a <SEP> 3 <SEP> x <SEP> lp / <SEP> lp <SEP> 10.00 <SEP> 134 <SEP> i <SEP> - <SEP>
<tb><SEP> Q3D <SEP> 13.30 <SEP> 152 <SEP><SEP> - <SEP> - <SEP>
<tb><SEP> 17.70 <SEP> 146 <SEP> - <SEP><SEP> 1/6
<tb> * LTS: Long-term survivor.

IV - STUDY OF TOXICITY COMPOUND ALGAE 21a
The substance to be tested is injected into mice
NMRI according to the protocol as specified in Table IV, below.

 After two weeks, the total number of deceased animals in each group is calculated and LD5a are evaluated using Litchfield Wilcoxon's method.

The results are shown in the Table
IV, hereinafter
TABLE IV

<tb> - <SEP> Substance <SEP> Program <SEP> Administration <SEP> DL50 <SEP>
<tb><SEP> 21a <SEP> 3 <SEP> x <SEP> ip <SEP>><SEP> 21.3
<Tb>
The anthracycline-containing drugs according to the present invention are generally administered at doses of from 0.001 to 25 mg / kg / day.

They are particularly suitable for the treatment of acute and chronic leukemias,
Hodgkin, with non-Hodgkin lymphoma.

 As is apparent from the above, the invention is not limited to those of its embodiments, implementation and application which have just been described more explicitly; it encompasses all the variants that may come to the mind of the technician in the field, without departing from the scope or the scope of the present invention.

Claims (12)

 at. said glycal of Formula 2 is converted into a halogenated derivative by adding a compound of formula HX anhydrous, wherein X is a halogen, to obtain a compound of Formula 19, according to scheme IV  2 ') Process for the preparation of new anthracyclines from a glycal according to claim 1, characterized in that R represents an optionally substituted, linear or branched alkyl group or an optionally substituted aryl group, with the exception of a CH2OH group or a CH20R3 group, R3 having the same meaning as R:  in which

 1 ') Glycals obtained from L-Fucose and / or its analogues according to Claim 1 of Formula 2

SCHEMA IV Koenigs-Knorr.  b. then said compound of Formula 19 is coupled to a suitable anthracyclinone, by a reaction of the type R 1 represents an acetyl group or a hydrogen atom. R is as defined above  in which

 3) Process according to claim 2, characterized in that said compound 19 is coupled with domeomycin and provides an anthracycline of Formula 20 '

 4) Process according to claim 2, characterized in that said compound 19 is coupled with B rhodomycinone and provides an anthracycline of Formula 21 '.  wherein R and R 1 have the same meanings as before.

R 1 represents an acetyl group or a hydrogen atom R 4 represents a hydrogen atom or a radical of Formula 19 ', R is as defined above in which,  5) Process according to any one of claims 2 to 4, characterized in that the glycosides obtained are purified by chromatography. R is as defined above R1 represents an acetyl group or a hydrogen atom.  in which

 6 ') Anthracycline, characterized in that it is obtained by the process according to claims 2 to 5 and in that it is of Formula 20', hereinafter  wherein R and R 1 have the same meanings as before.

R4 represents a hydrogen atom Qu a radical of For mule 19 ', R 1 represents an acetyl group or a hydrogen atom R is as defined above  in which,

 7 ') Anthracycline, characterized in that it is obtained by the process according to claims 2 to 5 and in that it is of Formula 21', hereinafter

O-acetyl-3 '' -4 'dideoxy-2', 6 '- alpha-L-threohexopyranosyl) -7,10 ss-Rhodomycinone of Formula 22a, hereinafter.  8) Anthracycline according to claim 7, characterized in that it is constituted by Di-O- (di

Rhodomycinone of Formula 21a, hereinafter  9) Anthracycline according to claim 7, characterized in that it is constituted by O- (di-O acetyl-3 ', 4' dideoxy-2 ', 6' - alpha-L-threo-hexopyranosyl) -7 ss-

 10-) Anthracycline according to claim 7, characterized in that it is constituted by Di-O- (di-O-acetyl-3,1,4 'trideoxy-2', 6 ', 7' - &alpha; -L-threo heptopyranno syl) -7,10 B-Rhodomycinone of Formula 22b, hereinafter  11) An anthracycline according to claim 7, characterized in that it is constituted by O- (di-O acetyl-3 ', 4' trideoxy-2 ', 6', 7 '- alpha-L-threo- heptopyranosyl) - 7 ss-Rhodomycinone of Formula 21b, hereinafter

O-acetyl-4'-desamino-3 'methyl-6-methyl-6'daunorubicin of Formula 20b, hereinafter  14 ') Anthracycline according to claim 6, characterized in that it consists of acetoxy-3

 13-) Anthracycline according to claim 7, characterized in that it is constituted by O- (di-O acetyl-3 ', 4' hexadésoxy-2 ', 6', 7 ', 8', 9 ', 10 '- &alpha;; - L-théodécapy-rannosyl) -7 B-Rhodomycinone of Formula 21c, hereinafter

 12) Anthracycline according to claim 7, characterized in that it consists of di-O- (di O-acetyl-3 ', 4' hexadésoxy-2 ', 6', 7 ', 8', 9 ', 10 '- &alpha; L-threo decapyranosyl) -7,10 ss-Rhodomycinone of Formula 22c, hereinafter

O-acetyl-4'-desamino-3'-propyl-6 'daunorubicin of Formula 20c, hereinafter  15 ') Anthracycline according to Claim 6, characterized in that it consists of 3'-acetoxy  17 ') A pharmaceutical composition characterized in that it contains as an active compound a turkey anthracy according to any one of claims 6 to 15, optionally combined with at least one pharmaceutically acceptable vehicle. R 1 and R 2 are the same or different and represent an acetoxy group or a hydrogen atom; R3 having the same meaning as R R represents an optionally substituted, linear or branched alkyl group or an optionally substituted aryl group, with the exception of a CH 2 OH group or a CH 2 O 3 R 3 group,  in which

 16 ') Intermediate for the preparation of glycals according to claim 1 of Formula 18