NZ203047A

NZ203047A - Daunorubicin and doxorubicin analogues and pharmaceutical compositions

Info

Publication number: NZ203047A
Application number: NZ203047A
Authority: NZ
Inventors: S Penco; G Franchi; F Arcamone; A M Casazza
Original assignee: Erba Farmitalia
Priority date: 1982-01-26
Filing date: 1983-01-18
Publication date: 1985-10-11
Also published as: NO153456B; PT76113A; FR2520365A1; ES519255A0; FI830155A0; NO830234L; SE461591B; BE895687A; ES8403140A1; GB2116169A; NO153456C; DK27683D0; FI74977B; DE3301489C2; GB2116169B; GB8301178D0; AU1052583A; FI74977C; FI830155L; IL67709A

Description

<div class="application article clearfix" id="description"> New Zealand Paient Spedficaiion for Paient Number £03047 20304 HO DRAWINGS Priority Date(s): Complete Specification Fifad: ~'~S3 c,sss: ' Publication Date- B ^ 19851 PO. Journal, No: ... /£76 NEW ZEALAND PATENTS ACT, 1953 No.: Date: COMPLETE SPECIFICATION DAUNORUBICIN AND DOXORUBICIN ANALOGUES, THEIR PREPARATION AND USE THEREOF S/We, FARMITALIA CARLO ERBA S.p.A., an Italian Company of Via Carlo Imbonati 24, 20159 MILAN (ITALY) hereby declare the invention for which XV we pray that a patent may be granted to osus/us, and the method by which it is to be performed, to be particularly described in and by the following statement:- - 1 - 2 03047 The present invention in one aspect thereof provides anthra cycline glycosides of the formula I wherein X is hydrogen or hydroxy, : is hydrogen or methyl, one of R2 and is methoxy and the other of Rg. and R^ hydrogen; and pharmaceutically acceptable acid additions salts thereof. These compounds 'are named as follows: -3- 203047 4-cemethoxy-4'-O-methy1-daunorubicin (Ta: R^R'-^X^H, R2=OCH3) , 4-demethoxy-4 *-epi-4'-O-methyl-daunorubicin (rb: r =r2=x=h, r3=och3), 4-demethoxy-2, 3-dimethyl-41-O-methy1-daunorubicin (Ic: R1=CH3, R2=OCH3, R3=X=H), 4-demethoxy-2,3-dimethy1-4'-epi-4'-O-methyl-daunorubicin (Id: R1=CH3, R2=X=H, R3=OCH3)7 4-demethoxy-4'-O-methy1-doxorubicin (Ie: R1=R3=H, R2=OCH3, X-OH), 4-deme t'noxy-4 ' -epi-4 ' -O-methy 1-doxorubicin (Ifr R1=R2=H, R3=OCH3, X=OH), 4-demethoxy-2,3-dimethy1-4'-O-methy1-doxorubicin (Ig: R1=CH3# R2=OCH3, R3=H, X=OH), and 4-demethoxy-2,3-dimethy1-4'-epi-4'-O-methy1-doxorubicin (Ih: R1=Ch*3/ R2=H, R3=OCH3, X=OH) . In another aspect thereof the invention provides a process for preparing the ant'nracycline glycosides of the formula I. The process according to the invention comprises condensing 4-demethoxy-daunomycinone or 2,3-dimethy1-4-demethoxy-daunomycinone (United States Patent No. 4046878) with 2,3,6-trideoxy-3-trifluoroacetamido-4-O- X' ^ ^ ^ ^ 'V X' ir,ethyl-L-lyxo-hexopyranosyl chloride or 2 , 3, 6-trj ceoxy/J. ^ OA ii -3-trif luoroace t ami do- 4-O-me thvl-L-arab ino-hexoDyranosfrl*. * . -n> '4UG a chloride (New Zealand Patent Specification No.. 186047) to give one of the protected-a-glycosides of the formulae Ila to lid, , ^ 2 03 047 Ila: r1=r3=h; r2=och3 lib: r!=r2=h; r3=och3 lie: VCH3'- R2=OCH 3; ui II lid: VCH3; R2=H; R3: =och3 NHCOCF. removing the N-trifluoroacetyl protecting group by mild alkaline hydrolysis to give the corresponding cauno-rubicin derivative la to Id, and optionally converting the daunorubicin derivative to the corresponding doxorubicin derivative le to Ih by bromination and treatment of the resulting 14-brorao-derivative with aqueous sodium formate. The conversion of the daunorubicin derivatives la to Id to the corresponding doxorubicin derivatives Ie to Ih follows the method described in United States Patent No. 3803124. • < In still another aspect thereof the invention provides pharmaceutical compos itions comprising an anthracycline glycoside of the formula I or a pharmaceutically acceptable salt thereof in admixture with a pharmaceutically acceptable diluent or carrier therefor. -5- 2030 The invention is illustrated by the followirig Examples and biological data. Example 1. 4-deme thoxy-4 ' -O— me thyl-'dauno rub 1 c 1 n (la) A solution of 3.6 8 g of 4-demethoxy-daunomycinone in 400 ml of anhydrous methylene dichloride containing 1.4g of 1-chloro-4-O-methy1-N-tri fluoroacety1-daunosamine was vigorously stirred in the presence of molecular sieve (30 g, 4A Merck) and 1.3g of silver trifluoromethane-sulphonate. After 10 minutes at room temperature, the reaction mixture was neutralized with 0.55 ml of sym-collidine. After 40 minutes the suspension"was filtered and the organic phase was washed with O.Ol N aqueous solution of hydrochloric acid, with water, with a saturated aqueous solution of sodium bicarbonate and finally with water to neutrality. The residue, obtained by evaporating off the solvent under vacuum, was dissolved in 150 ml of acetone, treated with 600 ml of 0.2 N aqueous sodium hydroxide and diluted with 450 ml of water. After 5 hours at 0°C the solution was adjusted to pH 8.5 and extracted with chloroform until the chloroform extracts were no longer colo red. The organic extracts were combined and acidified to pH 5 with O.l N methanolic hydrogen chloride. By evaporation under vacuum to a small volume (150 ml) pure 4-demethoxy-4'-O-methyl-daunorubicin (0.6 g) crystallized. The mother liquor was purified on a 203047 column of silica gel buffered at pH 7 with phosphate buffer M/15, using the solvent system chloroform:methanol: water (10:2:0.2 by volume). The eluate, containing the pure compound, was diluted with water and the organic phase was separated off, washed with water and evaporated to a small volume and thereafter acidified to pH 5 with 0.1 N methanolic hydrogen chloride. A further amount (0.4 g) of 4-demethoxy-41-0-methyldaunorubicin hydrochloride was obtained; m.p. 189-190°C (with decomposition). TLC on Kieselgel plates (Merck F 254) solvent system chloroform:met"nanol:water (10:2:0.2 by volume). EPLC: experimental analysis: column microbondapack mobile phase: water:acetonitrile (69:31 by volume) at pH 2 with 10% orthophosphoric acid: flux rate 1.5 ml/min., retention time 22 min. FD-MS: m/z 511 (M+*) Example 2 '4-demethoxy-4'* -0-methy1-doxorubicin (Ie) A solution of 0.5 g of 4—demethoxy-4'-0-methy1-aauno--rubicin, prepared as described in Example 1, in a mixture of methanol (8 ml)and dioxan (20 ml) was treated with bromine to form the 14-bromo derivative. Treatment of the 14-bromo derivative with an aqueous solution of sodium formate at room temperature for 18 hours gave O.310 g of 4-demethoxy-4'-0-methyl-doxorubicin which was isolated as 203047 its hydrochloride m.p. 164-165°C (with decomposition); TLC on Kieselgel plate (f'erck F 254) solvent system chloroform:methanol:water (10:2:0.2 by volume); Rf 0.18. HPLC: experimental conditions: column microbondapack mobile phase water:acetonitrile (69:31 by volume) at pH 2 with 10£ orthophosphoric acid; flux rate 1.5 ml/min; retention time; 10 min. ExamDle 3 *■ 4-demethoxy-2,3-dimethy1-4'-Q-methy1-daunorubicin (Ic) The coupling reaction between 4-demethoxv-2,3-dimethyl--daunomycinone and l-chloro-4-0-methy1-N-trifluoroacety1-daunosamine under the conditions described in Example 1 afforded the title compound. Example 4 4-demethoxv-2 , 3-dimet'nyl-4 ' -0-methyl-doxorubi"cin . (Ig) The conversion of compound Ic to the title compound was performed using the procedure described in Example 2. Example 5 4-demethoxy-4'-epi-4'-0-methyIdaunorubicin(lb) and 4--demethoxy-2,3-dime thyI-4'-eoi-4'-0-methy1-daunorubicin (Id) The coupling reactions, as described in Example 1, of 4— -demethoxy-daunomycinone and 4-cemethoxy-2,3-dimethyl--daunomvcinone with 2,3,6-trideoxy-3-trifluoroace tajnido-4--0-methy1-L-arabino-hexopyranosv1 chloride afforded, after hydrolysis of the N-protecting group, the title compounds. Example 6 4-demethoxy-4'-epi-4'-O-methy1dodorubicin (If) and 4-methoxy-2 , 3-dine thyl-4 ' -epi-4 ' -Q-rr.ethyl-doxorubicin (Ih) The compounds lb and Id were converted, via their 14-brono- -8- 2030 derivatives under the conditions described in Example 2 to compounds If'and Ih respectively. BIOLOGICAL' ACTIVITY OF Ta "and Ie The compounds la and ie were tested against the parent compounds, respectively daunorubicin (DNR) and doxorubicin. (DX), in several experimental systems, in order to ascertain their cytotoxicity, antitumour acitivity and cardiac toxicity in experimental animals. Data reported in Table 1 show that la is about 5 times more cytotoxic than DNR, and Ig; is about 9 times more cytotoxic than DX. The primary screening in vivo was carried out in CDF-1 mice bearing P388 ascitic leukemia (10^ cells/mouse). Results are reported in Table 2. Both la and Ie were found to be more toxic and more potent than the parent compounds. Comparison at the Maximal Tolerated Dose (MxTD) shows that la is as effective as DNR (giving similar increase of the mice life span) and Ie has a good antitumour activity, which is of the same order of magnitude, as that of DX. Several studies were carried out in C3H mice bearing the Gross leukemia injected i.v. (2x10^ cells/mouse). Data on la are reported in Table 3. Administered i.v. on day 1 after the tumour inoculation, la was markedly more toxic and more potent than DNR. At the MxTD of 1.25- -1- 2030 1.3 mg/Kg, la was more effective than DX at the MxTD of l 10 mg/Kg. It is well known that DNR is not active when administered by oral route, unless very high doses are given (^-50 mg/Kg) . Data reported in Table 3 show that la has good antitumour activity against Gross leukemia also when given orally. Administered orally on day 1 only, la is active at the dose of 1.25-1.3 mg/Kg, v;hich is also the optimal dose in the case of the i.v. treatment. This result suggests that absorption of la through the gastrointestinal tract is very efficient. Administered on days 1, 2 and 3 by oral route, la is more active than when administered on day 1 only; at the optimal dose of 0.6 6 mg/I'g/day it has an antitumour activity of the same order of magnitude as that of 4-deme thoxy-daunorubicin, which was investigated in parallel at the optimal dose of 1.9 mg/Kg/day. The observation that the optimal dose of la is lower than that of 4-demethoxy-daunorubicin suggests a more efficient absorption through the gastrointestinal tract also in comparison to this compound. Data on the antitumour activity of Ie in comparison with DX against Gross leukemia are reported in Table 4. The compounds were administered on day 1 after the tumour cells inoculum; DX was given i.v.; Ie was given i.v. and orally. When administered i.v., at the MxTD of 1 mg/Kg, Ie showed a good antitumour activity, similar to that observed after DX treatment. In addition, ie - 1.G- 20304 was active also when administered orally at doses from 1.3 mg/Kg on. Ie was tested against L 1210 leukemia, which was inoculated in CDF-1 mice i.p. (ascitic form) or i.v. (lO^ cells/mouse). Treatment was performed on day 1 after the tumour cell inoculation, i.p. or i.v., respectively. Data reported in Table 5 show that, in the i.p.-i.p. experiment, Ie at the MxTD of 0.83 mg/Kg was as active DX at the MxTD of 4.4 mg/Kg. In the i.v.-i.v. experiment, Ie at the tolerated doses of 1 and 1.3 mg/Kg showed antitumour activity superior to that of DX. In order to assess the antitumour activity against a solid tumour, Ie was tested in comparison with DX, against the mammary carcinoma of C3H female. A third generation tumour transplant was utilized. Treatment started 15 days after the tumour transplant, and was performed i.v. once a week for 4 weeks. Tumour measurement was performed by caliper every week. Non-tumour bearing mice were treated in parallel, in order to evaluate the general toxicity and the cardiac toxicity, which was investigated in 5 mice, treated with the highest doses of the two compounds, and killed 5 weeks after the last treatment. The results of this experiment are reported in Table 6. DX was very effective, and it inhibited tumour growth by 93-94% (in comparison with the untreated controls) at both the doses tested (6 and 7.5 mg/Kg). In the non-tumoured mice treated with DX at _11 _ 2030 7.5 mg/Kg, toxic deaths were observed (2/3) and all the mice examined showed histologically detectable heart lesions. Ie at the dose of 0.4 mg/Kg was slightly active; at the doses of 0.6 and 0.75 mg/Kg it markedly inhibited the tumour growth. In non-tumoured mice treated with Ie at O.75 mg/Kg no atrium lesions were observed, and only 2 out of 5 mice showed detectable ventricle lesions, uhicn were less severe than those observed after DX treatment. The data here presented show that la and Ie are new anthracycline analogues endowed with very interesting biological properties. In comparison with DNR, la is about 3 times more potent when administered i.p., and about 8 times more potent when administered i.v. At the MxTD it has an antitumour activity against ascitic P388 and systemic Gross leukemia equal to that of DNR. In addition, it is active also when administered by oral route, particularly when treatment is performed for 3 consecutive days, at doses lower than those of 4-demetnoxy--daunorubicin. Ie is about 10 times more potent than DX in vivo. Comparison at the MxTD shows that it is, in respect to DX, equally active against ascitic P 388 and L 1210 leukemia, systemic Gross leukemia and solid mammary carcinoma, and it is more effective than DX against systemic (i.v. injected) L 1210 leukemia. In addition, ib active against -12- 2 03 0 47 Gross leukemia also when administered orally, and in a preliminary cardiotoxicity test in C3H mice treated chronically i.v. it caused only minimal cardiac lesions. Compound la has .been studied on P388. leulcemia cells resistant to doxorubicin (P388/DX) in vitro and vivo. P388 leukemia cells resistant to doxorubicin (DX) (received by dr. Schabel) are maintained by serial transfer in mice treated with DX i.p.. Lf For experimental purpose, BDF-1 mice were injected with 10 leukemia cells i.p., and treated i.p. on day 1 after the tumor inoculation. Results, reported in Table 7, show that daunoru bicin (DNR) was not active against this tumor, while compound la, at the optimal dose of 0.8 mg/kg increased the life span of the treated mice. P388 and P388/DX leukemia cells were harvested from mice ascitic fluid and adapted to grow in suspension in vitro. Cytotoxicity tests were carried out exposing the cells to vaious drug concentrations for 48 hrs; at the end of the exposure period, cells were counted with a Coulter Cell Counter, and the ID.--, (dose which gives 50% reduction of the cell number du in comparison with untreated controls) was calculated. Results, reported in Table 8, show that la was about twice as cytotoxic as DNR on P388 leukemia cells, and was very active also on P388/DX leukemia cells, while DNR was 163-152 times less active on the resistant than on the sensitive line. Compound Ie was further investigated on mammary carcinoma of it d C3H mice. Mice bearing measurable tumor (3 generation transplant) were treated once/week for 4 weeks i.v. with Ie or with DX. Normal mice were treated in parallel, for evalua _13 _ 20304 tion of toxicity. Results, reported in Table 9, confirm that Ie was about 10 times more potent than DX, and had a remarkable antitumor activity in this experimental system at non toxic doses, while DX was toxic. Because of the good antitumor activity against mammary car cinoma, compound Ie was tested against two solid tumors: the colon 26 and the colon 38 adenocarcinomas, transplanted s.c. in BALB/c mice and in BDF-1 mice, respectively. Treat ment started- on day 1 after the tumor inoculation (early) or when tumor was already palpable (advanced), and was per formed i.v. once/week or every 6 days, for 3 or 4 times. Tumor growth was assessed by caliper measurement. Non-tumor bearing mice were treated in parallel, for toxi_ city evaluation, and were observed for 90 days. Results of three experiments are reported in Table 10. Against early colon 26, Ie at the maximal tolerated dose of 0.9 mg/kg/day was more active than DX at the maximal tolerated dose of 7.5 mg/kg/day. Against advanced colon 26, Ie at the maximal dose tested of 0.7 mg/kg/day was not toxic, ana gave a higher tumor growth inhibition than. DX at the maximal tolerated dose of 6 mg/kg/day. Against advanced colon 38, Ie at the maximal dose tested of 0.9 mg/kg/day was as active as DX 9 mg/kg/day in inhibiting tumor growth, but produced a higher increase of survival time. In conclusion, all data here reported confirm that compounds la and Ie are extremely interesting new anthracyclines, endowed with high potency, activity by oral route, activity 2030 14 - against anthracycline resistant tumors. (la), activity against solid tumors superior to that of DX and not cardioto xic (Ie) . Table 1 - Colony inhibition test against Hela cells in vitro (Treatment for 24 hrs) Compound Dose (ng/niT)' ' (ng/tal) DNR 12.5 42 6.2 66 ^ 12 3. 1 121 la 25 0 6.2 O ^2.5 1.5 73 DX 12.5 20 6.2 79 % 9- 3.1 177 Ie 10 0 2.5 9.9 ^ 1 0.62 82 O. 15 84 ^o. of colonies; % of untreated controls. -15- 203047 Table 2. Antitumour activity against P388 leukemia Treatment i.p. on Day 1 Compound Dose .'(mg/Kg). . . T/C3 " • ' % LTSb. Toxic ' ' deathsc DN Rd 2.9 15 4 O/lO O/10 4.4 140,154 0/20 4/20 6.6 109,163 0/20 13/20 la 0.5 12 7 0/10 O/lO 0.64 127 0/10 0/10 0.8 172,127 0/20 1/20 l.O 145 O/lO O/lO 1.3 95 O/lO 10/10 DX 4 . 4 180 O/lO O/lO 6.6 200 2/10 O/lO 10. oe 315 4/10 0/10 Ie 0.44 180 0/10 O/lO 0.66 215 O/lO O/lO 1.0 250 0/7 1/7 1.5 245 2/10 4/10 1 a Median survival time; % over untreated controls Long term survivors (>60 days) c Evaluated on the basis of autoptic findings on dead mice ^ Data of two experiments e This is the Maximal Tolerated Dose of DX in this experimental system 203047 -Ik- Table 3 . Activity of la against Gross leukemia Treatment ' T/C b Route. Schedule.3 Compound mg/kg/day Toxi c . deaths0 i - v. 1 DNR 10 15 22.5 133,150 175,175, 203,191, 166 216 0/20 3/30 6/30 i .v. oral JL 1 la 0.58 0. 76 1.0 1.25-1.3 1. 56 1. 95 1. 1 1.25-1.3 1.56-1.6 1.95-2.0 116 133 158,166 183,208 100 116 133 . 133,166 166,166 175,183 O/IO O/lO 0/20 0/20 9/10 6/10 O/IO 1/20 3/20 2/2 O oral 1,2,3 4-demethoxy DNR 1. 31 1.58-1.46 1. 9 2.5 3.3 125 133,150 133,190 210 140 O/IO 1/20 1/20 2/10 6/10 oral 1,2,3 la 0. 44 0.66 1.0 1.25 133 183 200,220 140 0/9 0/10 9/20 8/10 a _ _ Days after tumour inoculation h, c see Table 2. 2 03 0 -i?- - Table 4. Activity of Ie against Gross leukemia Treatment - T/Cb Toxic c Route Compound ' ' _ mg/kg ■% deaths i. v. DX 10 200 2/8 13-- 200,216 2/18 16.9 250,266 3/18 Ie l.O 183,233 0/18 1.3 192,208 4/18 1.7 217,200 7/18 2.2 142 7/10 oral Ie l.O 125 0/8 1.3 150 0/8 1. 7 166 0/8 2.5 166 n / a <J/ o. . . On Day 1 after tumour inoculation t>,c See Table 2. Table 5. Activity of Ie against L12 10 leukemia L1210 inoculum Treatment Compound mg/kg - T/C3 % LTSb Toxic ,. c ceatns i.p. i.p. DX 4.4 150 0/9 0/9 6.6 150 0/10 1/10 10.0 162 O/IO 1/10 l€. 0. 83 162 2/10 0/10 1.0 187 l/lO 1/10 . 1.2 393 4/10 3/10 i.v. i.v. DX 10.0 120 O/IO 0/10 13.0 120 0/10 0/10 16.9 133 O/IO 0/10 • ie- 1.0 200 0/10 0/10 1.3 173 O/IO 0/10 1.7 >580- 5/10 0/10 ■ a,b,c See Table 2. Table 6. Activity against C3H mammary carcinoma, toxicity and cardiac toxicity of Ie in comparison with DX mg/kg/day Tunoured mi.ce • Hon tumoured mice deaths Heart lesions ]Compound Tumour growth % Tumour growth, inhibition % T/C Compound - A V • MST • %' n. ■ G ' n'. G - - 4845 - 53 - - - - - ! DX 6 318 93 77 145 n. d. i 1 7.5 277 94 74 140 2/3 » 4/4 1.3 5/5 .1.5 j 1 Ie 0. 4 1740 64 65 12 3 n. d. 0.6 748 85 76 143 n. d. 0/5 0 2/5 0.2 0.75 3 4 4 9 3 81 15 3 1/4 i a Tumour weight on day 4 3/tumour weight at beginning of treatment, x 100. ^ 100~/Tumour growth of treated mice/tumour growth of controls, X•1007 C Median survival time (days) ^ Observed for 90 days 6 A=atrium; V=ventricles; n.=number of heairts showing lesion/total; G : lesions grade Is) O u o - 19 - 2 03 04 7 Table 7. Effect on doxorubicin-resistant P388 leukemia in vivo. Compound Dose (mg/kg) T/C3 % LTSb Toxic Q deaths DNR 2.9 93 0/10 0/10 4.4 87 0/10 1/10 6.6 84 0/10 3/10 la 0.53 133 0/10 0/10 0.8 142 1/10 0/10 1.2 106 0/10 4/10 3Median Survival time; % over untreated controls. ^Long-term survivors (> 60 days) Q Evaluated on the basis of autoptic findings on dead mice - 20 - 203047 Table 8. Effect on sensitive and doxorubicin-resistant P388 leukemia in vitro. r a ID50 (ng/ml)b e Compound P388c P388/DXd DNR 5.8,2,3 950,350 163,152 la 0.35,1.3 1.4,5 4,3.8 aData of two experiments. ^Dose giving 50% reduction of cell number in comparison with untreated controls. c P388 leukemia cells sensitive to DX ^P388 leukemia cells resistant to DX eRatio between IDcn on P388/DX and IDcn on P388 bU bU Table 9 Activity of Ie against mammary carcinoma of C3H mice Compound mg/kg/day Tumor weight (nig) Tumored T/Cb % mi ce MSTC T/Cd % Non tumored mice deathse - - 651 115.5 0/10 DX 6.0 238 37 149 129 1/10 7.5 209 32 100 87 7/10 Ie 0.6 422 65 118 103 0/10 0.75 263 40 126.5 110 0/10 Evaluated 1 week after the last treatment, by caliper measurement. ^Tumor weight of treated mice/tumor weight of controls, x 100. £ Median survival time (days). ^MST of treated mice/MST of controls (x100). e Mice were observed for 90 days. </div>

Claims

<div class="application article clearfix printTableText" id="claims"> - 22 - 203047 Table 10 Effect of Ie and DX against two transplanted colon adenocarcinomas in mice Tumor Stage3 Schedule'3 Compd. Dose (mg/kg) %C inhib. T/Cd % Toxi c deaths6 Colon 26 early q7dx4 DX 6 56 217 0/10 7.5 82 224 0/10 9.3 81 161 9/10 Ie 0.6 81 258 0/10 0.75 85 227 0/10 0.9 93 246 1/10 advanced q6dx3 DX 6 34 237 0/9 9 62 237 4/9 Ie 0.6 48 222 0/9 0.7 52 195 0/9 Col on 38 advanced q7dx4 DX 6 65 92 0/10 9 83 144 1/10 Ie 0.6 55 133 0/10 0.75 60 129 0/10 0.9 81 184 0/10 Time of start of treatment in respect to tumor development. 3Days of i.v. administration. "% inhibition of tumor growth, as compared with untreated controls. "'Median survival time of treated mice/median survival time of controls, x 100. "Evaluation in non-tumored mice treated in parallel and observed for 90 days. 2030^7 - 23 ~ What we claim is: 1) Anthracycline glycoside compound having the formula I: wherein X is hydrogen or hydroxy; R is hydrogen or methyl; one of R2 and R^ is methoxy and the other of R2 and hydrogen and pharmaceutically acceptable acid addition salts thereof." 2) A compound according to claim 1, which is 4-demethoxy--4'-0-methy1-daunorubicin. 3) A compound according to '-'claim 1, which is 4-demethoxy-_4 « -0-oie thy 1 -doxorubicin . 203047 - - 4) A compound, according to claim. 1, which, is 4-demethoxy-4 r-epi-4 f-O-methyl-daunorubicin. 5) A compound according to claim 1, which is 4-demethoxy-41 --epi-4r-O-methyl-doxorubicin. 6)A compound according to claim 1, which is 4-demethoxy-2,3-dimethyl-4'-O-methyl-daunorubicin. 7) A compound according to claim.r.1 , which is 4-demethoxy--2,3-dimethy1-4' -G -methyl-doxorubicin. 8) A compound according to claim 1, which is 4-demethoxy--2,3-dimethyl-AT-epi-4'-O-methyl-daunorubicin . 9) A compound according to claim 1, which is 4-demethoxy--2,3-dimethyl-4'-epi-4 r-O-methyl-doxorubicin. 10) A process for preparing the anthracycline glycosides according to claim 1, comprising reacting 4-demethoxy-daunomycinone or 2,3--dimethyl-4-demethoxy-daunomycinone, dissolved in anhydrous methylene dichloride with 2,3,6-trideoxy-3-tri f'luoroacetamido-4-0-methyl-L-lyxo-hexopyranosy1 chloride or 2,3,6-trideoxy-3-trifluoroacetamido 4-0-methyl-L-arabino hexopyranosyl chloride, in the presence of silver trifluoro methane-sulphonate and molecular sieve, to obtain the cor responding M-trifluoroacetyl protected ^-glycosides, removing the N-trifluoroacetyl protecting group by mild alkaline hydrolysis with 0.
2 M aqueous sodium hydroxide to obtain the corresponding daunorubicin derivatives (I:X=H) which <20 ?C>(f7 203047 -K- are optionally converted to the corresponding doxorubicin derivatives (I:X = OH) by bromination and treatment of the thereby formed 14-bromo-intermediates with aqueous sodium for mate. 11) A pharmaceutical composition comprising a therapeutically effective amount of an anthracycline glycoside as claimed in claim 1 in combination with an inert carrier therefor. 12) A method of inhibiting the growth of a tumor, in a non-human animal, j selected frcm the group consisting of P 388 leukemia, Gross leukemia, L 1210 leukemia and mammary carcinoma comprising administe ring to a host afflicted therewith, a therapeutically effecti ve amount of anthracycline glycoside as claimed in claim 1. 13) A protected glycoside of the formula: NHCOCF. wherein R is hydrogen or methyl, R^ is hydrogen or-met;hoxy and R2 is hydrogen or methoxy provided that R2 and R^ are not simultaneously the same. 203047 2O*Jo4-7 26 14. Anthracycline glycoside compound as claimed in claim 1 and substantially as herein described with reference to any of the examples. 15. A process for preparing the anthracycline glycosides as claimed in claim 10 and substantially as herein described with reference to any of the examples. 16. A pharmaceutical composition as claimed in claim 11 and substantially as herein described with reference to any of the examples. 17. A method of inhibiting the growth of a tumor as claimed in claim 12 and substantially as herein described with reference to any one of the examples. 18. A protected glycoside as claimed in claim 13 and substantially as herein described with reference to any of the examples. PER AGENTS FOR THE APPLICANTS </div>