NEW 4'-EPI-4'-AMINO ANTHRACYCLINES

The present invention relates to a new class of anthracycline glycosides having antitumour activity, methods for their preparation, pharmaceutical compositions containing them and the use thereof in treating certain mammalian tumours. The invention also relates to the preparation of certain novel intermediates.

The invention provides anthracycline glycosides having the general formulae 1 _^ and 2 \

wherein R^ is selected from the group consisting of hydrogen, fluorine, hydroxy and amino; 2 a ^{nd R} 3 both represent hydroxy or one of R ₂ and R ₃ is hydrogen, nitro or amino and the other of R ₂ and R. is hydroxy; and

pharmaceutically acceptable salts thereof. Preferred salts are the hydrochloride salts.

The anthracycline glycosides of the general formulae 1_ and ,2 include:

4-demethyl-3'-deamino-4'-deoxy-4'-epi-amino-daunorubicin (la: R ₁ =R ₂ =R- _J =OH)

4-demethyl-3'-deamino-4'-deoxy-4'-epi-amino-doxorubicin (2a: R ₁ =R ₂ =R ₃ =OH)

4-demethoxy-4-amino^3'-deamino-4*-deoxy-4'-epi-a ino- daunorubicin (lb: ₁ =NH ₂ , R ₂ =R ₃ =OH)

4-demethoxy-4-amino-3'-deamino-4'-deoxy-4'-epi-amino- doxorubicin (2b: R ₁ =NH ₂ , R ₂ =R ₃ =OH)

4-demethoxy-4-fluoro-3*-deamino-4'-deoxy-4*-epi-amino- daunorubicin (lc: R^F, R ₂ =R ₃ =OH)

4-demethoxy-4-fluoro-3'-deamino-4'-deoxy-4'-epi-amino- doxorubicin (2c: R^F, ₂ = ₃ =OH)

4-demethyl-6-deoxy-3'-deamino-4 ^, -deoxy-4'-epi-amino- daunorubicin (Id: R ₁ =R ₃ =OH, R ₂ =H)

4-demethyl-6-deoxy-3'-deamino-4'-deoxy-4'-epi-amino- doxorubicin (2d: R ₁ = ₃ =OH, R ₂ =H)

4-demethoxy-ll-deoxy-ll-nitro-3'-deamino-4'-deoxy-4'-epi- amino-daunorubicin (le: R^H, R ₂ =OH, R ₃ =N0 ₂ )

4-demethoxy-ll-deoxy-ll-nitro-3'-deamino-4'-deoxy-4'-epi- amino-doxorubicin (2e: R ₁ =H, R ₂ =OH, R ₃ =N0 ₂ )

4-demethoxy-6-deoxy-6-nitro-3'-deamino-4'-deoxy-4'-epi- amino-daunorubicin (If: R ₁ =H, R ₂ =N0 ₂ , R.,=OH)

4-demethoxy-6-deoxy-6-nitro-3*-deamino-4*-deoxy-4'-epi- amino-doxorubicin (2f: R-^H, R ₂ =N0 ₂ , R ₃ =0H)

4-demethoxy-ll-deoxy-ll-amino-3*-deamino-4'-deoxy-4'-epi- amino-daunorubicin (lg: R- H, R ₃ =NH ₂ , R ₂ =OH)

4-demethoxy-ll-deoxy-ll-amino-3'-deamino-4'-deoxy-4'-epi- amino-doxorubicin (2g: R-^H, R ₃ =NH ₂ , R ₂ =OH)

4-demethoxy-6-deoxy-6-amino-3'-deamino- '-deoxy-4'-epi- amino-daunorubicin (lh: ^H, R ₂ =NH ₂ , R ₃ =OH)

4-demethoxy-6-deoxy-6-amino-3'-deamino-4'-deoxy-4'-epi- amino-doxorubicin ⁽ 2h: R ₁ =H, R ₂ =NH ₂ , ₃ =OH ₎

The new anthracycline glycoside antibiotics of the invention, i.e. those of formula 1 and 2, are condensation

products of (a) aglycones of general formula 3 _^ or 6 _^ :

wherein R, , R ₂ and R ₃ are as defined above with proviso that neither R~ nor R, is an amino group and (b) a protected halosugar of the formula 4^

wherein X is a halogen, preferably chlorine.

The synthetic methods for the preparation of the above mentioned anthracyclines follows two methods: Method A is depicted in Scheme I and involves the use of the aglycone of formula 2. ₁ ^an - Method B, which is shown in Scheme II and involves the use of the aglycone of formula 16. The preparation of the C-6 or C-11 amino compounds of formula 2 _^ is illustrated in Scheme III.

Scheme I

3a-f

5a-£ la-ϊ

1 ) Br m, /άioxane-methanol 2 )HCCONa

2a-f

Scheme II

3a-f 6a-f

7a-f Ba-f

9a-f

i: Br ₂ /dioxane, CH- _j COOK; ii: 4_, CF- _j SO-Ag, CH ₂ C1 ₂ ; iii: K ₂ C0 ₃ , CH- _j OH, 0°C; iv: HC(OC ₂ H ₅ ) ₃ , pTSA-pyridine v: 0.1N NaOH, CH- _j COOH

Scheme III

9e,£

1) 10%Pd/C cyclohexene-methanό reflux 10 min.

2) 0.2N NaOH

3) CH ₃ COOH

2g,h

Method A

The present invention provides a process for the preparation of an anthracycline glycoside of formula 1 or 2 as defined above with proviso that for a glycoside of formula 2 neither ₂ nor R ₃ is an amino group, or a pharmaceutically acceptable acid addition salt thereof, which process comprises:

(i) condensing an aglycone of formula 3:

wherein R,, R- and R_ are as defined above except that neither R ₂ nor R-, is an amino group, with a l-halo-2,3,4,6- tetradeoxy-4-(N-trifluoroacetamido)-L-erythro-hexopyranoside of formula 4:

wherein X is halogen;

(ii) removing the N-trifluoroacetyl group from the compound of formula 5 thus obtained:

wherein R, , R ₂ and R ₃ are as defined in step (i), so as to obtain a said anthracycline glycoside of formula 1 except that neither R ₂ nor ₃ is an amino group;

(iii) if desired, converting the said glycoside of formula 1 obtained in step (ii) into a pharmaceutically acceptable acid addition salt thereof;

(iv) if desired, reducing a said glycoside of formula 1 wherein one of R ₂ and R ₃ is a nitro group obtained in step (ii) or a said salt thereof obtained in step (iii) so as to obtain a said glycoside of formula 1 wherein one of R ₂ and R ₃ is an amino group and, if desired, converting the said glycoside of formula 1 wherein one of ~ and R» is an amino group into a pharmaceutically acceptable acid addition salt thereof;

(v) if desired, brominating the said glycoside of formula 1 obtained in step (ii) or pharmaceutically acceptable acid addition salt thereof obtained in step (iii)

and hydrolysing the 14-bromo derivative thus obtained to form a corresponding anthracycline glycoside of formula 2 as defined above; and

(vi) if desired, converting the said glycoside of formula 2 into a pharmaceutically acceptable acid addition salt thereof.

This procedure allows the preparation of anthracycline glycosides from the corresponding aglycones 3_ and a protected halosugar 4_. The procedure is similar to that described in US-A-4,107,423. The coupling product is hydrolyzed to a daunorubicin derivative 1_ and can be converted to the corresponding doxorubicin derivative 2 _^ in accordance with the method described in US-A-3,803,124.

The starting materials for the reaction sequences of Scheme I are the well known 4-demethyldaunomycinone (3a_:R-=R ₂ =R ₃ =OH) , the aglycone 4-demethoxy-4-amino- dauno ycinone (3_ : ₁ =NH ₂ , R ₂ =R ₃ =OH) (EP-A-0288268) , 4-demethoxy-4-fluorodaunomycinone (3£: R,=F, R ₂ =R ₃ =OH) [G.W. Morrow and J. Swenton, J. Org. Chem. ; 52 _^ , 713, 1987] and 4-demethyl-6-deoxydaunomycinone (3<3: R,=R ₃ =OH, R =H) [US-A-4,600,537] . The C-11 and C-6 nitro aglycones 4-demethoxy-ll-deoxy-ll-nitrodaunomycinone (^e: R,=H, R ₂ =OH, R ₃ =N0 ₂ ) and 4-demethoxy-6-deoxy-6-nitrodaunomycinone (3f: R =H, R =N0 ₉ , R ₃ ^=0H ) both described in US-A-4,749,693.

An aglycone of formula 3 is generally reacted at room temperature in step (i) with a compound of formula 4_ in the

presence of a molecular sieve and silver trifluoromethane- sulphonate to form one of the N-trifluoroacetyl glycosides 5a-f. Compounds of formula la-f are obtained by removing the amino protecting group by mild alkaline hydrolysis. Preferably in step (ii) the compound of formula 5, dissolved in acetone, is submitted, at a temperature of 0°C and for one hour, to mild alkaline hydrolysis with 0.2 N aqueous sodium hydroxide to give a said glycoside of formula 1. Treatment with methanolic hydrogen chloride yields the hydrochlo ide.

In step (iv), typically reduction is effected by treatment with Pd/C, for example 10% Pd/C. The nitro glycoside of formula 1 or salt thereof may therefore be refluxed in a mixture of methanol and cyclohexene in the presence of 10% Pd/C for ten minutes. The resulting amino glycoside of formula 1 can be isolated as its hydrochloride as above.

Preferably the said glycoside of formula 1, dissolved in a mixture of anhydrous methanol and dioxane, is treated in step (v) with a chloroform solution of bromine to afford the corresponding 14-bromo derivative which is hydrolyzed for two days at room temperature with an aqueous solution of sodium formate to give a said glycoside of formula 2 as the free base and in step (vi) the said glycoside of formula 2 is isolated as its hydrochloride. Isolation of the glycoside of formula 2 as the hydrochloride is typically

achieved by treating the glycoside with methanolic hydrogen chloride.

According to an embodiment of this process 4-demethyldaunomycinone (3a) , dissolved in dry ethylene chloride, is reacted at room temperature for one hour with l-chloro-2,3,4,6-tetradeoxy-4-(N-trifluoroacetamido)-L- erythro-hexopyranoside (4_) in presence of molecular sieves and silver trifluoromethanesulphonate to obtain the N-protected glycoside ^a which, dissolved in acetone, is submitted at a temperature of 0 C and for one hour, to a mild alkaline hydrolysis with 0.2N aqueous sodium hydroxide to give the compound of formula JLa_ as a free base which, by treatment with anhydrous methanolic hydrogen chloride is isolated as its hydrochloride.

If desired, ^_ - ^s reacted with bromine in methylene chloride to obtain its 14-bromo derivative from which, after hydrolysis at room temperature and for 48 hours under nitrogen with an aqueous solution of sodium formate, the compound of formula J2a is obtained as free base and, by treatment with anhydrous methanolic hydrogen chloride, is isolated as its hydrochloride.

Method B «

The invention provides a process for the preparation of an anthracycline glycoside of formula 2 as defined above, with the proviso that neither ₂ nor R ₃ is an amino group, or a pharmaceutically acceptable acid addition salt thereof, which process comprises:

(i') condensing a 14-protected aglycone of formula 6:

wherein R, , R ₂ and ₃ are as defined above except that neither R ₂ nor R ₃ is an amino group, with a l-halό-2,3,4,6- tetradeoxy-4-(N-trifluoroacetamido)-L-erythro-hexopyranoside of formula 4 as defined above;

(ii') removing the 14-protecting group from the resulting N-protected glycoside of formula 7:

wherein R, , R ₂ and R ₃ are as defined above, to give the compound of formula 8:

wherein R,, R ₂ and R ₃ are as defined above;

(iii') converting the compound of formula 8 into a 9,14-orthoformate derivative of formula 9:

wherein R _j , R ₂ and R ₃ are as defined above;

(iv*) removing the N-trifluoroacetyl group and the orthoformate protecting group to obtain a said glycoside of formula 2; and

(v ¹ ) if desired, converting the said glycoside of formula 2 into a pharmaceutically acceptable acid addition salt thereof.

This procedure allows the preparation of anthracycline glycosides of general formula 2_ by coupling (a) the corresponding 14-hydroxylated and protected aglycones and (b) a protected halosugar Jk The procedure is similar to that described in US-A-4,107,423. In Scheme II is depicted the route to doxorubicin derivatives 2a-f. The starting materials are the aglycones 3a-f suitably functionalized and protected in the side chain as 14-acetoxy derivatives.

Typically in step (i') the aglycone of formula 6 is reacted at room temperature with a 1-chloro-hexapyranoside of formula 4 in the presence of a molecular sieve and silver trifluoromethansulfonate. Preferably in step (ii ¹ ) the compound of formula 7 is treated at 0 C, for four hours and under a nitrogen atmosphere with potassium carbonate. Generally in step (iii') the compound of formula 8 is treated with triethyl orthoformate, for example in methylene chloride and in the presence of pyridinium p-toluensulfonate for one hour at room temperature.

Preferably in step (iv ¹ ) the compound of formula 9 is subjected to mild alkaline aqueous hydrolysis to remove the N-trifluoroacetyl group and is treated with acetic acid to remove the orthoformate protecting group to obtain the said glycoside of formula 2 as a free base which, in step (v ¹ ), is treated with anhydrous methanolic hydrogen chloride to isolate the glycoside as its hydrochloride salt.

More particularly in one embodiment 4-demethoxy-ll- deoxy-11-nitrodaunomycinone (3d: R,=H, R ₂ =OH, R_=N0 ₂ ) is dissolved in dry dioxane and treated with a solution of bromine in methylene chloride for two hours at room temperature, then precipitated with hexane. The residue, dissolved in acetone, is added with potassium acetate and stirred for one hour to give the acetoxy derivative 6d. This compound is dissolved in dry methylene chloride and •reacted with l-chloro-2,3,4,6-tetradeoxy-4-(N-trifluoro- acetamido)-L-erythro-hexopyranoside (4_) as described above to give the protected 14-acetoxy-N-trifluoroacetyl glycoside 7d.

In order to remove the protecting groups, compound 73 is first dissolved in methanol and treated with potassium carbonate for four hours at 0°C under nitrogen to give 8d, then is treated with triethyl orthoformate in methylene chlo¬ ride and in the presence of pyridinium p-toluensulfonate for one hour at room temperature to give the 9,14-orthoformate derivative 9e _^ . This is subjected to mild alkaline aqueous hydrolysis to remove the N-protecting group and finally

treated with acetic acid to remove the orthoformate protecting group and give compound le_ as free base'which, by treatment with anhydrous methanolic hydrogen chloride, is isolated as its hydrochloride.

The present invention also provides a process for the preparation of an anthracycline glycoside of formula 2 above wherein R. is as defined above and one of R ₂ and ₃ is hydroxy and the other of R ₂ and R ₃ is an amino group, or a pharmaceutically acceptable acid addition salt thereof, which process comprises:

(i ⁿ ) reducing the C-6 or C-11 nitro group of a 9,14-orthoformate derivative of formula 9 as defined above wherein one of R ₂ and R ₃ is hydroxy and the other of R_ and R ₃ is a nitro group;

(ii") removing the N-trifluoroacetyl group and the orthoformate protecting group from the C-6 or C-11 amino group-containing compound thus formed to obtain a said glycoside of formula 2; and

(iii") if desired, converting the said glycoside of formula 2 into a pharmaceutically acceptable acid addition salt thereof.

Scheme III shows the route to C-6 and C-11 amino anthracyclines of formula 2 starting from the corresponding C-6 and C-11 protected nitro glycosides. The 9,14-ortho- formate nitro derivatives j3e_ or 9_f may be converted respectively to amino glycosides _2g_ or _2h _. by refluxing the

nitro compounds in a mixture of methanol and cyclohexene in the presence of 10% Pd/C for ten minutes, and removing the N-trifluoroacetyl group in a basic medium, for example as described above for Method A or Method B, and the orthoformate protecting group with acetic acid. The compounds J2 and J ^m -Y be converted into their respective hydrochloride salts by treatment with anhydrous methanolic hydrogen chloride.

As is apparent from the foregoing, the processes of the invention involve the preparation and use of several novel intermediates. These too are within the scope of the present invention, more especially the compounds of formulae 5 and 7 to 9.

The invention also provides pharmaceutical compositions comprising an anthracycline glycoside of formula 1 or 2 or a pharmaceutically acceptable acid addition salt thereof, together with a pharmaceutically acceptable carrier or diluent. Conventional carriers and diluents may be used. The composition may be formulated and administered in conventional manner.

The compounds of the invention are useful in methods of treatment of the human or animal body by therapy. They are useful as antitu or agents. A therapeutically effective amount is administered to a patient. An amount sufficient to inhibit the growth of the tumour may be administered. The tumour may be a Colon adenocarcinoma or Gross leukemia

tumour .

The biological activity of the compounds according to the invention was tested iτ\ vitro against LoVo (human colon adenocarcinoma) and LoVo/DX cells in comparison with doxorubicin and 4-demethoxydaunorubicin (4-dem-DNR). The results are shown in Table 1.

Table 1 - i_n vitro activity

Compounds Cytotoxicity after 4h of treatment

(IC50 ¹ - ng/ml) LoVo LoVo/DX R.I. ²

1) IC50 - concentration inhibiting by 50% colony growth

2) R.I. « Resistance Index = (IC50 LoVo/DX)/(IC50 LoVo)

The following Examples illustrate the invention

Example 1 Preparation of

4-demethyl—3'-deamino-4'-deoxy-4'-epi-amino-daunorubici n hydrochloride (la)

0.76 g (2 mmole) of 4-demethyldaunomycinone (3_a) was dissolved with 300 ml of anhydrous methylene chloride in the presence of molecular sieves (4 ) . The mixture was cooled at 10 ^β C, bubbled with nitrogen and added dropwise, under stirring, with 0.86 g (3.2 mmole) of l-chloro-2,3,4,6-tetra- deoxy-4-(N-trifluoroacetamido)-L-erythro-hexopyranoside (4_) dissolved in 40 ml of anhydrous methylene chloride and 0.78 g (3.0 mmole) of silver trifluoromethanesulphonate dissolved in 40 ml of diethyl ether. After twenty ir.inutes the reaction mixture was treated with 30 ml of aqueous saturated, sodium hydrogen carbonate and filtered off. The organic layer was washed with water, dried over anhydrous sodium sulphate and the solvent was removed in vacuo. The residue was chromatographed over a silica gel column using a mixture of methylene chloride acetone (98:2 by volume) to give 0.8 g of 4-demethγl-3'-deamino-4'-deoxy-4'-epi-N- trifluoroacetyldaunorubicin (5_a) . The trifluoroacetyl protecting group was removed by dissolving compound 5_a in acetone and treating with aqueous 0.2N sodium hydroxide at 0°C. After one hour the solution was adjusted to pH 8.1 and extracted repeatedly with methylene chloride. The combined organic extracts, after being dried and concentrated to a small volume were acidified to pH 3.5 with anhydrous methanolic hydrogen chloride. Upon addition of diethyl ether there was obtained 0.64 g (yield 54%) of the title compoun la as hydrochloride salt. m.p. 168-169°C (with dec). TLC on Kieselgel Plate (Merck ^F 254^ solvent system: methylene chloride/methanol/acetic acid/water (80:20:7:3 b volume) Rf:0.73. MS-FD: [M]+ 497.

^X HNMR (200 MHz, DMSO-dg) δ:

1÷21 (d, J=6.2Hz, 3H, δ'-CH- _j ), 1.6-1.8 (m, 4H, 2'-CH ₂ , 3'-CH ₂ ), 2.1-2.3 (m, 2H, 8-CH ₂ ), 2.27 (s, 3H, COCH- _j ), 2.85 (m, 1H, 4 ^f -H), 2.95 (m, 2H, 10-CH ₂ ), 4.11 (dq, J=6.2, 9.9Hz, 1H, 5'-H), 4.98 (m, 1H, 7-H) , 5.22 (s, 1H, 9-OH) , 7.40 (dd, J=2.0, 7.5Hz, 1H, 3-H), 7.82 (m, 2H, 1-H, 2-H) , 8.10 (bs, 3H, 4'-NH ₃ +), 11.96 (s, 1H, 11-OH) , 12.87 (s, 1H, 6-OH) , 13.40 (s, 1H, 4-OH)

Example 2

Preparation of

4-demethoxy-4-amino-3'-deamino-4' -deoxy-4'-epi-amino-dauno¬ rubicin hydrochloride (lb)

0.38 g (1 mmole) of 4-demethoxy-4-aminodaunomycinone (3b) was coupled with 0.37 g (1.5 mmole) of l-chloro-2,3,4,6-tet deoxy-4-(N-trifluoroacetamido)-L-erythro-hexopyranoside (_4) following the procedure described in Example 1 to affor

0.35 g (yield 60%) of the title compound lb as hydrochlorid salt.

TLC on Kieselgel Plate (Merck ∑ solvent system methylene chloride/methanol/acεtic acid/water (80:20:7:3 b volume) Rf:0.45

¹ HNMR (200 MHz, DMSO-dg) δ (inter alia):

2.27 (s, 3H, COCH-.), 2.85 (m, 1H, 4'-H), 4.98 (m, 1H, 7-H)

6.80 (bd _f 2H, 4-NH ₂ ) , 6.93 (d, J=8.0Hz, 1H, 3-H) , 7.46 (t

J=8.0Hz, 1H, 2-H), 7.64 (d, J=8.0Hz, 1H, 1-H) , 8.10 (bs, 2H

4'-NH ₃ +), 13.52 (s, 1H, 11-OH) , 14.00 (s, 6-OH)

Example 3

Preparation of

4-demethoxy-4-fluoro-3'-deamino-4'-deoxy-4'-epi-amino-dau no rubicin hydrochloride (lc)

0.38 g (1 mmole) of 4-demethoxy-4-fluorodaunomycinone (3c was coupled with 0.37 g (1.5 mmole) of l-chloro-2,3,4,6-tet deoxv-4-(N-trifluoroacetamido)-L-erythro-hexopyranoside (4) following the procedure described in Example 1 to affor

0.48 g of N-trifluoroacetyl derivative 5_c.

After removal of the N-protecting group, following standar procedure, was obtained 0.40 g (yield 74 %) of the titl compound lc.

TLC on Kieselgel Plate (Merck ^F 254^ solvent system methylene chloride/methanol/acetic acid/water (80:20:7:3 b volume) Rf:0.68, m.p. 158-159°C (with dec.)

Example 4

Preparation of

4-demethyl— '-deamino-4'-deoxy-4 -epi-amino-doxor bicin hydrochloride (2a)

0.3 g (0.6 mmole) of 4-demethyl-3'-deamino-4'-deoxy-4' -epi- aminodaunorubicin (la) was issolved in a mixture anhydrous methanol and dioxane and added with 1.2 ml of solution containing 9 g of bromine in 100 ml of methyle chloride, according to the method described in US-A-3,803, 124, to afford the 14-bromo derivative, which wa dissolved in 20 ml of acetone and treated with 0.4 g sodium formate dissolved in 2 ml of water. The - reacti mixture was stirred at room temperature for two days, th water was added and extracted with methylene chloride. Aft standard work up, the resulting ^" red solution w concentrated to small volume

under vacuum, adjusted to pH 3.5 with anhydrous methanolic hydrogen chloride, then added with an excess of diethyl ether to give 0.2 g, (yield 75%), of the title "compound 2a as hydrochloride.

TLC on Kieselgel Plate (Merck solvent system: methylene chloride/methanol/acetic acid/water (80:20:7:3 b volume) Rf:0.60

Example 5 Preparation of 4-demethyl-6-deoxy-14-acetyl-adriamycinone (6d)

To a solution of 0.85 g (2.3 mmole) of 4-demethyl-6-deoxy- daunomycinone (3_d) , dissolved in 150 ml of anhydrou dioxane, was added 4.6 ml of solution containing 9 g o bromine in 100 ml of methylene chloride. The mixture wa left at room temperature for two hours, after that the bro derivative was precipitated .by-adding n-hexane and collecte

The residue was dissolved with 300 ml of acetone and adde under stirring with 2.7 g of potassium acetate. After on hour the reaction mixture was diluted with methylen chloride and washed with water. The organic layer was drie over anhydrous sodium sulphate, filtered off .and the solven was removed under reduced pressure. The residue wa chromatographed on a column of silica gel to give 0.75

(yield 76%) of the title compound 6d.

TLC on Kieselgel Plate (Merck F-e _* ) solvent system methylen chloride/acetone (95:5 by volume) Rf:0.13

EI-MS: [M]+ 426

¹ HNMR (200 MHz, DMSO d _g ) δ:

1.88 (dd _r J=9.3, 12.1 Hz, 1H, 8ax-H) , 2.06 (s, 3H, COCE,)

2.41 (dd, J=5.7, 13.1 Hz, 1H, 8e-H) , 2.76-3.04 (two d,

J=18.6 Hz, 2H, 10-CH ₂ ), 4.57 (m, 1H, 7-H) , 5.09-5.23 (two d, J=17. ^' 8 Hz, 2H, COCH ₂ OCO), 5.89 (d, J=6.6 Hz, 1H, 7-OH) , 6.10 (s, 1H, 9-OH), 7.39 (dd, J=1.5, 7.9Hz, 1H, 3-H) , 7.7-7.9 (m, 2H, 1-H, 2-H), 7.90 (s, 1H, 6-H) , 12.60-12.90 (two bs, 2H, 4-OH, 11-OH) .

Example 6

Preparation of

4-demethyl-6-deoxy-3'-deamino-4'-deoxy-4'-epi-N-trifluoac et- yl-doxorubicin (8d)

0.61 g (1.43 mmole) of 4-demethyl-6-deoxy-14-acetyl-adria- mycinone (6ιd) was dissolved with 250 ml of anhydrous methylene chloride and 150 ml of anhydrous tetrahydrofurane. The mixture was cooled at 10°C, bubbled with nitrogen and added dropwise, under stirring, with lg (4 mmole) of l-chloro-2,3,4,6-tetradeαxγ-4-(N-trifluoroacetamido)-L- ervthro-hexopyranoside (4^ dissolved in 50 ml of anhydrous methylene chloride and 1 g (4 mmole) of ^' silver trifluoro ethanesulphonate dissolved in 50 ml of diethyl ether. After twenthy minutes the reaction mixture was treated with 20 ml of aqueous saturated sodium hydrogen carbonate and filtered off. The organic layer was washed with water, dried over anhydrous sodium sulphate and the solvent was removed in vacuo. The residue was chromatographed over a silica gel column to give 0.4 (yield 44%) of 4-demethyl-6-deoxγ-14-acetyl-3'-deamino-4' - deoxγ-4'-epi-N-trifluoroacetyldoxorubicin (7d) . TLC on Kieselgel Plate (Merck F-, ₅₄ ) solvent system methylen chloride/acetone (9:1 by volume) RF:0.45 FD-MS: [M]+ 636

^Η MR (200 MHz, DMSo ^' dg) δ, (inter alia):

1.06 (d, J=6.0 Hz, 3H, 5'-CH ₃ ), 1.6-2.0 (m, 5H, 8ax-H,

2'-CH ₂ , 3'-CH ₂ ), 2.05 (s, 3H, COCH ₃ ) 3.56 (m, 1H, 4'-H),

3.93 (m, 1H, 5'-H), 5.14 (m, 2H, COCH ₂ OCO) , 5.17 (m, 1H, l'-H), 9.37 (bd, J=8.6Hz, 1H, NHCOCF ₃ ), 12.45, 12.91 (two s,

2H, 4-OH, 11-OH).

Compound 7d was dissolved with 200 ml of methanol, treated at 0°C with 2 ml of a 10% aqueous solution of potassium carbonate and left to stand for four hours at 0°C under nitrogen.

The solution was neutralized with acetic acid, diluted with water and the product extracted with methylene chloride.

After silica gel filtration 0.25 g of the title compound 8d was recovered (yield 89%) .

TLC on Kieselgel Plate (Merck solvent system: methylene chloride/acetone (9:1 by volume) Rf:0.26.

FD-MS: [M]+ 593.

Example 7 Preparation of

4-demethyl-6-deoxy-3' -deamino-4' -deoxy-4' -epi-amino-doxorub¬ icin, hydrochloride (2d)

0.2 g (0.33 mmole) of product 8d, prepared as described i Example 6, was. dissolved with 60 ml of dry methylene chloride and added with 15 ml of triethyl orthoformate an 0.1 g of pyridinium p-toluensulfonate. After one hour th solution was washed with water, dried over anhydrous scdiu sulphate and reduced to small volume under reduced pressure. The solution was poured into n-hexane and the precipitate o 9,14-orthoformate derivative 9d was collected by filtration. TLC on Kieselgel Plate (Merck ^F 254^ solvent system: methylene chloride/acetone (95:5 by volume) Rf:0.7

The residue was dissolved with 200 ml of 0.2N aqueous sodium hydroxide and the solution was left for ten hours at 8°C under nitrogen. Then the solution was adjusted at pH 8.5 with acetic acid and extracted with methylene cloride. After standard work up, the resulting free base was treated with an aqueous solution of acetic acid for two hours at room temperature. The mixure was brought to pH 8.5 with aqueous sodium hydrogen carbonate and extracted with methylene chloride. The solvent was removed in vacuo and the residue chromatographed on silica gel column to afford 0.12 g (yield 66%) of the title compound as free base which was transformed into the hydrochloride salt 2ά by treatment with anhydrous methanolic hydrogen chloride. TLC on Kieselgel Plate (Merck F ₂₅₄ ) solvent system: methylene chloride/methanol/acetic acid/water (80:20:7:3 by volume) Rf:0.52. m.p. 155°C (dec.) FD-MS: [M3+ 43B ^X HNMR (200 MHz, DMSO d _g ) δ:

1.21 (d, J=6.2Hz, 3H, 5'-CH ₃ ), 1.7-2.0 (m, 5H, 8ax-H, 2'-CH ₂ , 3'-CH ₂ ), 2.4-2.5 (m, 1H, 8e-H) , 2.77, 3.08 (two d, J=17.7Hz, 2H, 10-CH ₂ ), 2.88 (m, 1H, 4'-H), 3.95 (dq, J=6.2, 9.8Hz, 1H, 5'-H), 4.55 (d, J=.5.5Hz, 2H, CH ₂ OH) , 4.76 (m, 1H, 7-H), 4.84 (t, J=5.5Hz, 1H, CH ₂ OH) , 5.20 (m, 1H, l'-H), 5.93 (s, 1H, 9-OH), 7.40 (dd, J=1.6, 7.7Hz, 1H, 3-H) , 7.7-7.9 (m, 2H, 2-H, 1-H) , 7.74 (s, 1H, 6-H) .

Example 8 Preparation of 4-demethoxy-ll-deoxy-ll-nitro-14-acetyladriamycinone (6e)

2.3 g (5.7 mmole) of 4-demethoxy-ll-deoxy-ll-nitro-daunomy- cinone (3_e) was transformed into 1.8 g, (yield 70%), of the title compound 6je following the procedure reported i Example 5. TLC on Kieselgel Plate (Merck ₂₅₄ ) solven system: methylene chloride/acetone (95:5 by volume) Rf:0.2. FD-MS: [M]+ 455. ¹ HNMR (200 MHz, CDC1 ₃ ) δ:

2.11 (ddd, J=1.8, 5.0, 15.0Hz, 1H, 8ax-H) , 2.18 (s, 3H COCH ₃ ), 2.56 (ddd, J=2.2, 2.2, 15.0Hz, 1H, 8e-H) , 2.88 (dd J=2.2, 18.2Hz, 1H, lOe-H) , 3.18 (d, J=18.2Hz, 1H, lOax-H) 3.42 (dd, J=1.8, 3.3Hz. 1H, 7-OH) , 4.68 (s, 1H, 9-OH) , 5.04 5.34 (two d, J=18.2Hz, 2H, COCH ₂ OCO) , 5.42 (m, 1H, 7-H) 7.88 (m, 2H, 2-H, 3-H) , 8.29 (m, 2H, 1-H, 4-H) , 13.71 (s 1H, 6-OH).

Example 9 Preparation of ^• 4-demethoxy-ll-deoxy-ll-nitro-9,14-orthoformate-3'-deamino- 4'-deoxy-4'-epj-N-trifluoroacetyl-doxorubicin (9e)

1.76 g (3.8 mmole) of product 6_e was condensed with 1.3 (5.2 mmole) of the chloro-sugar _, 4 in presence of silve trifluoromethanεsulphonatε, following the procedur described in Example 1, to give 1.13 g (yield 44%) o 14-acetyl-N-trifluoroacetyl-derivative 7e.

TLC on Kieselgel Plate (Merck F ₂₅₄ ) solvent syste methylene chloride/acetone (95:5 by volume) Rf:0.37. FD-MS: [M]+ 665

¹ HNMR (200 MHz, CDCl- _j ) δ:

1 28 (d, J=5.8Hz, 3H, S'-CE^), 1.8-1.9 (m, 4H, 2'-CH ₂ , 3'-CH ₂ ), 2.10 (dd, J=3.8, 15.0Hz, 1H, 8ax-H) , 2.18 (s, 3H, COCH ₃ ), 2.56 (ddd, J=1.6, 1,6, 15.0Hz, 1H, 8e-H) , 2.94 (dd, J=1.6, 18.3Hz, 1H, lOe-H), 3.19 (d, J=18.3Hz, 1H, lOax-H) , 3.7-4.1 (m, 2H, 4'-H, 5'-H), 4.94, 5.32 (two d, J=17.8Hz, 2H, COCH ₂ OCO), 5.40 (m, 2H, 7-H, l'-H), 6.55 (bd, J=8.6Hz, 1H, NHC=CF ₃ ), 7.87 (m, 2H, 2-H, 3-H) , 8.2-8.4 (m, 2H, 1-H, 4-H), 13.70 (s, 1H, 6-OH)

Product 7e was dissolved with 1000 ml of methanol and, after cooling at 0°C, a 10% aqueous solution of sodium carbonate was added under stirring and nitrogen atmosphere. After threε hours, the mixture was brought to pH 7 with acetic acid, diluted with water and the product was extracted with methylene chloride following standard procedure. The crude material was crystallized from diethyl ether to give 0.9 g (yield 87%) of compound 4-demethoxy-ll-deoxy-ll-nitro-3'- deamino-4'-deoxy-4'-epi-N-trifluoroacetyl-doxorubicin (8e) TLC on Kieselgel Plate (Merck F ₂₅₄ ) solvent system: methylene chloride/acetone (95:5 by volume) Rf:0.17. Product 8e was treated with triethylorthoformate in presence of py.ridinium p-toluensulfonate as described in Example 7 to give 0.78 g (yield 76%) of the title compound 9e. TLC on Kieselgel Plate (Merck F ₂₅₄ ) solvent system: methylene chloride/acetone (95:5 by volume) Rf:0.48.

Example 10 Preparation of

4-demethoxy-ll-deoxy-ll-nitro-3'-deamino-4'-deoxy-4'-epi- amino-doxorubicin (2e)

Following the procedure described in Example 7, 0.32 g (0.43 mmole) of compound 9_e was first hydrolyzed in basic media to remove the N-protecting group then with acetic acid to remove the orthoformate protecting group. Treatment with anhydrous methanolic hydrogen chloride afforded 0.16 g (yield 61%) of the title compound 2_e as hydrochloride with TLC on Kieselgel Plate (Merck F ₂₅₄ ) solvent system: methylene chloride/methanol/acetic acid (80:20:1 by volume) Rf:0.30. m.p. 159°C (dec.) MS-FD: [M]+ 527 ¹ HNMR (200 MHz, DMSO d _g , 50°C) δ:

1.23 (d, J=6.2Hz, 3H, 5'-CH ₃ ), 1.7-1.9 (m, 4H, 2'-CH ₂ , 3'-CH ₂ ), 2.26 (m, 2H, 8-CH ₂ ), 2.82 (m, 2H, lO-CI ), 2.84 (m, 1H, 4'-H), ^' 4.16 (dq, J=6.2, 9.5Hz, 1H, 5'-H), 4.4-4. (m, 3H, Cθgi ₂ OH, COCH ₂ OH) , 5.08 (m, 1H, 7-H) , 5.24 ( , 1H, l'-H), 5.53 (s, 1H, 9-OH), 7.98 (m, 2H, 2-H, 3-H) , 8.1-8. (m, 2H, 1-H, 4-H).

Example 11

Preparation of

4-demethoxy-ll-deoxy-ll-amino-3'-deamino-4'-ceoxy-4' -epj- amino-doxorubicin hydrochloride (2g)

0.32 g (0.43 mmole) of 4-demethoxy-ll-deoxy-ll-nitro-9,14- ethyl-orthoformate-3'-deamino-4'-deoxy-4'-epi-N-trifluoroac etyldoxorubicin (9_e) , prepared as described in Example 9 was dissolved in 200 ml of methanol and added with 20 ml o cyclohexene and 0.2 g of 10% Pd/C, under stirring. Th mixture was refluxed for ten minutes, then cooled at room

temperature and the catalyst was filtered off and the solvent removed under vacuum. The ^' residue was picked up with aqueous 0.2N sodium hydroxide and kept for eight hours at •10 ^β C under nitrogen. After that, the solution was brought to pH 5 with acetic acid and left- to stand for two hours at room temperature. The mixture was neutralized with aqueous sodium hydrogen carbonate and extracted with methylen chloride to give, after standard work up, the 11-amin deivative as free base. Treatment of which with methanoli hydrogen chloride afforded 0.15 g (yield 55%) of the titl compound 2σ_. TLC on Kieselgel Plate (Merck F ₂₅₄ ) solven system: methylene chloride/methanol/acetic acid (80:20:1 b volume) Rf:0.23. m.p. 162-164 ^β C (dec.) FD-MS: [M]+ 497 ^X HNMR (200 MHz, DMSO dg, 50°C) δ:

1.23 (d, 3H, 5'-CH ₃ ) , 1.7-1.9 (m, 4H, '-CH ₂ , 3'-CH ₂ ), 2.21 (m, 2H, 8-CH ₂ ), 2.80 (m, 2H, 4'-H), 2.88 (m, 2H, 10-CH ₂ ), 4.18 (dq, J=6.2, 9.5Hz, 1H, 5'-H), 4.67 ( , 3H, COCH ₂ OH, COCH ₂ OH), 5.05 (m, 1H, 7-H) , 5.28 (m, 1H, l'-H) 5.31 (s, 1H, 9-OH), 7.88 (m, 2H, 2-H, 3-H) , 8.27 (m, 2H 1-H, 4-H) , 8.18, 8.40 (two bm, 4H, 11-NH ₂ , 4'-NH ₂ ).

Example 12 Preparation of 4-demethoxy-6-deoxy-6-nitro-14-acetyl-adriamycinone (6f)

2 g (5 mmole) of 4-demethoxy-6-deoxy-6-nitro-daunqmycinone

(3f) was transformed into 1.7 g (yield 75.6%) o£ 14-acetyl derivative 6f _, following the procedure described in

Example 5. TLC on Kieselgel Plate (Merck ₂₅₄ ) solvent system: methylene chloride/acetone (95:5 by volume) Rf:0.32.

FD-MS: [M]+ 455

¹ HNMR (200 MHz, CDCL ₃ ) δ:

2.12 (dd, J=14.9, 4.8Hz, 1H, 8ax-H) , 2.20 (s, 3H, OCOCH- _j ),

2.53 (ddd, J=2.0, 2.4, 14.9Hz, 1H, 8e-H) , 3.15 (d, J=19.3Hz,

1H, lOax-H), 3.38 (dd, J=2.0, 19.3Hz, 1H, lOe-H) , 3.43 (m,

1H, 7-OH), 5.00 (m, 1H, 7-H) , 5.15, 5.31 (two d, J=17.9Hz,

2H, COCH ₂ OCO), 7.89 (m, 2H, 2-H, 3-H) , 8.30 (m, 2H, 1-H,

4-H), 14.46 (s, 1H, 11-OH)

Example 13

Preparation of .»

4-demethoxy-6-deoxy-6-nitro-9,14-orthoformate-3'-deamino- 4'-deoxy-4'-epi-N-trifluoroacetyl-doxorubicin (9f)

1.7 g (3.7 mmole) of 4-demethoxy-6-deoxγ-6-nitro-14-acetyl- adriamycinone (6f_) was coupled with 1.6 g (6 mmole) of l-chloro-2,3,4,6-tetradeoxy-4-(N-trifluoroacetamido)-L- erythro-hexopyranoside (4.) in presence of silver trifluoromethanesulphonate as de- _^ scribed in^Example 1 to give 1 g (yield 41%) of N-trifluoroacetyl derivative 7_f_. TLC on Kieselgel Plate (Merck ₂₅₄ ) solvent system: methylene chloride/acetone (95:5 by volume) Rf:,Q.50. FD-MS: [M]+ 665

0.5 g (0.75 mmole) of compound 7L was treated with triethyl¬ orthoformate in presence of pyridinium p-toluensulfonate as described in Example 9 to afford, after chromatographic separation, 0.3 g (yield 60%) of the title compound 9f_ as mixture of two diastereoisomers with a molar ratio of 80/20. TLC on Kieselgel Plate (Merck F ₂₅ .) solvent system: methylene chloride/acetone (95:5 by volume) Rf:0.65. ^X HNMR (220 MHz, CDC1 ₃ ) δ ^'

1.1-1.3 (m, 6H, 5'-CH ₃ , OCH ₂ CH ₃ ), 1.6-2.0 (m, 4H, 2'-CH ₂ , 3'-CH ₂ ), 2.3-2.7 (m, 2H, 8-CH ₂ ), 3.33 (s, 2H, 10—CH ₂ ), 3.6-3.7 (m, 2H, OCH ₂ CH ₃ ), 3.75 (m, 1H, 4'-H), 3.90 (dq, J=6.2, 9.5Hz, 1H, 5'-H), 4.32, 4.45 (two d, J=17.4Hz, COCH ₂ major isomer), 4.20, 4.35 (two d, J=16.8Hz, COCH^O mino isomer), 4.87 (m, 1H, l'-H), 5.01, 5.14 (dd, J=5.3, 5.3Hz, 1H, 7-H major, 7-H minor), 5.70, 5.72 (s, 1H, CH-OCH ₂ CH major, CH-OCH ₂ CH ₃ minor), 6.04 (bd, J=9.0Hz, 1H, NHCOCF.^) 7.85 (m, 2H, 2-H, 3-H) , 8.27 (m, 2H, 1-H, 4-H) , 13.52, 13.5 (s, 1H, 11-OH major, 11-OH minor)

Example 14

Preparation of

4-demethoxy-6-deoxy-6-nitro-3 -deamino-4'-deoxy-4'-eoi-amin doxorubicin hydrochloride (2f)

0.3 g (0.44 mmole) of compound J9f, prepared as describe above, was dissolved in 200 ml of 0.2N aqueous sodiu hydroxide and kept for ten hours at 0 ^β C under nitrogen. Th solution was brought at pH 8.5 with acetic acid an extracted with methylene chloride. The aqueous solution wa adjusted to pH 8.1 and extracted repeatedly with methylen chloride. The combined organic extracts, after being drie and concentrated to a small volume were acidified to pH 3. with anhydrous methanolic hydrogen chloride.

- 33 - Upon addition of diethyl ether there was obtained 0.062 g (yield 25%) of the title compound 2f_ as hydrochloride salt. TLC on Kieselgel Plate (Merck F ₂₅₄ ) solvent system: methylene chloride/methanol/acetic acid/water (30:4:1:0.5 by volume) Rf:0.30 m.p. 155-157 ^β C (with dec.) MS-FD: [M]+ 527 ¹ HNMR (200 MHz, DMSO d _g ) δ:

1.18 (d, J=6.3Hz, 3H, 5'-CH ₃ ), 1.6-1.9 (m, 4H, 2'-CH ₂ , 3 ^f -CH ₂ ), 2.1-2.5 (m, 2H, 8-CH ₂ ), 2.84 (m, 1H, 4'-H), 3.01 (m, 2H, 10-CH ₂ ), 4.06 (dq, J=6.3, 9.3Hz, 1H, 5-H) , 4.53 (m, 2H, CH ₂ OH), 5.07 (m, 1H, 7-H) , 5.26 (m, 1H, l'-H), 5.50 (m, 1H, CH ₂ OH), 7.88 (m, 2H, 2-H, 3-H) , 8.14 (bs, 3H, 4'-NH ₃ +), 8.24 (m, 2H, 1-H) , 4-H)

Example 15 Preparation of

4-demethoxy-6-deoxy-6-amino-3'-deamino-4'-deoxy-4'-epi- amino-doxorubicin hydrochloride (2h)

0.15 g (0.22 mmole) of 9,14-orthoformate derivative 9f prepared as described in Example 13, was dissolved in 150 m of methanol and 15 ml of cyclohexene and treated with 0.15 10% Pd/C as describεd in Example 11 to give, upon additio with anhydrous methanolic hydrogen chloride, 0.02 (yield 18.5%) of the 6-amino derivativε 2h.

TLC on Kieselgel Plate (Merck F ₂₅₄ ) solvent system methylene chloride/methanol/acetic acid/water (30:4:1:0.5 b volume) Rf:0.22. MS-FD: [M]+ 497 ^X HNMR (200 MHz, DMSO dg, 50 ^β C) δ:

1.18 (d, J=6.3Hz, 3H, 5'-CH ₃ ), 1.6-1.9 (m, 4H, 2'-CH ₂ , 3 CH ₂ ), 2.1-2.5 (m, 2H, 8-CH ₂ ), 2.84 (m, 1H, 4'-H), 3.01 (m 2H, 10-CH ₂ ), 4.06 (bq, J=6.3, 9.3Hz, 1H, 5-H) , 4.53 (m, 2H CH ₂ OH), 5.07 (m, 1H, 7-H) , 5.26 (1H, l'-H), 5.50 (m, 1H CH ₂ OH), 7.88 (m, 2H, 2-H, 3-H) , 8.14 (bs, 3H, 4'-NH ₃ +), 8.2 (m, 2H, 1-H, 4-H)

Example 16

Preparation of 4-demethoxy-6-deoxy-6-nitro-3'-deamino-4'- deoxy-4'-epi-amino daunorubicin hydrochloride (If)

0.78 g (1.76 mmole) of 4-demethoxy-6-deoxy-6-nitro- daunomycinone (3f) was coupled with l-chloro-2,3,4,6- tetradeoxy-4-(N-trifluoroacetamido)-L-erythro-hexopyranoside (4) following the procedure described in Example 1 to afford 0.26 g (yield 62%) of the title compound If as hydrochlo ide salt.

TLC on Kieselgel plates (Merck ₂₅₄ ) solvent system: methylene chloride/methanol/acetic acid/water (80:20:7:3 by volume) Rf * 0.42. MS-FD [M]+ 511 ¹ H NMR (200 MHz, DMSO-d _g ) inter alia δ:

2.34 (s, 3H, COCH ₃ ); 3.10 (d, J=18.7 Hz, 1H, lOax-H) ; 3.27 (dd, J=1.8, 18.7Hz, 1H lOeg-HJ , 5.11 (dd, J=2.3, 4.3Hz, 1H, ^• 7-H) ; 7.8-7.9 (m, 2H, 2-H, 3-H) ; 8.10 (bs, 3H, 4'-NH ₃ +); 8.2-8.4 (m, 2H, 1-H, 4-H) ; 13.55 (ε, 1H, 11-OH) . Example 17

Preparation of 4-demethoxy-6-deoxy-6-amino-3'-deamino-4'- deoxy-4'-epi-aminodaunorubicin hydrochloride (lh)

0.3 g (0.5 mmole) of compound If was dissolved in 200 ml of methanol and 20 ml of cyclohexene and treated with 0.2 g of 10% Pd/C. After refluxing for ten minutes,the catalyst was filtered off and solvent removed in vacuo. After crystallisation from methanol/ethyl ether, 0.2 g (Yield 76%) of the title compound lh, as hydrochloride salt, was

obtained. TLC on Kieselgel plates (Merck F ₂₅₄ ) solvent system; methylene chloride/methanol/acetic acid/water (80:20:7:3 by volume) Rf=0.39. MS-FD: [M]+ 431 ^X H NMR (200 MHz, DMSO-d _g ) inter alia g:

1.18 (d, J=6.3 Hz, 3H, 5'-CH ₃ ); 1.6-1.9 (m, 4H, 2 ^» -CH ₂ , 3'-CH ₂ ); 2.1-2.5 (m, 2H, 8-CH ₂ ) ; 2.84 (m, 1H, 4'-H); 3.01 (m, 2H, 10-CH ₂ ); 4.06 (bq, J=6.3, 9.3Hz, 1H, 5-H) ; 5.7 ( , 1H, 7-H) ; 5.26 ( , 1H, l'-H); 7.8-7.9 (m, 2H, 2-H, 3-H) ; 8.10 (bs, 3H, 4'-NH ₃ +); 8.2-8.4 (m, 2H, 1-H,4-H). Example 18

Preparation of 4-demethoxy-ll-deoxy-ll-nitro-3'-deamino-4'- deoxy-4'-epi-amino-daunorubicin hydrochloride (le)

0.58 g (1.46 mmole) of 4-demethoxy-ll-deoxy-ll-nitro- daunomycinone (3e) was coupled with sugar (4) following the procedure described in Example l.to afford, after usual work up, 0.5 g (yield 61%) of the title compound le as hydrochloride salt.

TLC on Kieselgel plates- (Merck ₂₅₄ ) solvent system: methylene chloride/methanol/acetic acid/water (80:20:7:3 by volume) Rf = 0.44. MS-FD [M]+ 511 ^X H NMR (200 MHz, DMSO-d _g ) inter alia £: 1.18 (d, J=6-3Hz, 3H, 5'-CH ₃ ) ; 1.6-1.8 (m, 4H, 2'-CH ₂ , 3'-CH ₂ ); 2.1-2.5 (m, 2H, 8-CH ₂ ) ; 2.37 (s, 3H, COCH_ ₃ ) ; 3.07 (m, 2H, 10-CH ₂ ); 4.06 (bq, J=6.3, 9.3Hz, 1H, 5'-H); 5.03 (m, 1H, 7-H); 5.32 (m, 1H, l'-H); 7.8-7.9 (m, 2H, 2-H, 3-H) ;

8.10, bs, 3H, 4-NH ₃ +) ; 8.2-8.4 (m, 2H, IH, -IE) . 13.7 (s, IH, 6-OH) . Example 19

Preparation of 4-demethoxy-ll-deoxy-ll-amino-3'-deamino-4'- deoxy-4'-epiaminodaunorubicin hydrochloride (lg)

0.2 g (0.36 mmole) of compound le was transformed into its reduced amino derivative lg, folowing the procedure described in Example 17.

Yield 80%. TLC on Kieselgel plates (Merck F ₂₅₄ ) solvent system: methylene chloride/methanol/acetic acid/water (80:20:7:3 by volume) Rf = 0.41. MS-FD [M]+ 481.