IE871538L

IE871538L - Anthracycline glycosides

Info

Publication number: IE871538L
Application number: IE871538A
Authority: IE
Original assignee: Erba Farmitalia
Priority date: 1986-06-12
Filing date: 1987-06-10
Publication date: 1987-12-13
Also published as: CH676985A5; AU7410887A; DK169076B1; GB2195998A; BE1000158A4; FI84075B; PT85056A; JP2516769B2; GB8614323D0; FI84075C; ZA874202B; GB2195998B; CA1291122C; PT85056B; NZ220604A; ES2006488A6; AT392793B; KR950004897B1; ATA144987A; GB8713443D0

Abstract

This invention discloses new anthracycline glycosides having the general formula II: <IMG> II a,b wherein IIa, R1 represents a hydrogen atom and in IIb, R1 represents a hydroxyl group, and their pharmaceutically acceptable acid addition salts. These compounds are useful antitumor agents. [CA1291122C]

Description

The invention relates to novel anthracycline glycoside derivativese to their preparation and to pharmaceutical compositions containing them™ The present invention provides anthracycline glycosides having the general formulae (II)s CH,0 OH O r°H H^ypo II *.b a: n3 ~ H b: = OH wherein E1 represents a hydrogen atom or hydroxy1 group; and pharmaceutical^ acceptable acid addition salts thereof.

The invention also provides a process for tn preparation of a glycoside of formula (II) wherein R.( represents a hydrogen acort.? i.e. compound (Ila) , or a hydroxy grcip, i .e. compound (lib) , or a phar aaceuticall acceptable acid addition salt thereof, which process comprises converting 3e-dea«iino«4'-deoxy-3 f-epi-4'-epi-3', 4f-epimino-daunorubicin of formula (la ) : into the corresponding N-trifluoroacetyl derivative; converting the said N-tri£luoroacetyl derivative into 4'~ deoxv-4"-epi~tr ifluoroacetamido~3®-deamino~38-bydroxy-daunorubicin; removing the N-trifluoroacetyl group from the 4"-deoxy-4 °-epi-trifluoroacetaraido-3 9 -dearnino-3 '-hydroxy-daunorubicin so as to obtain the glycos > of formula (II) wherein R is a hydrogen atom? if desired, converting the said glycoside of formula (II) into a pharmaceutically acceptable acid addition salt thereof; if desired, brominating the said glycoside of formula (II) or pharmaceutically acceptable salt thereof and hydroly?i»jg the 14-bromo derivative thus obtained so as to form the glycoside of formula (XI) wherein is a hydroxy group; and, if desired, converting the said glycoside formula (II) wherein Hx is a hydroxy group into a pharmaceutically acceptable acid addition salt thereof.

COCH3 O - 3 - Treatment of the 3%, 4'-epimino daunorubicin •s derivative (la) with trifluoroacetic anhydride gives the j corresponding N-trifluoroacetyl derivative (Ve)» Reaction of this compound with a catalytic 5 amount of sulfuric acid in acetone gives 43-deoxy-4a-epi-tr ifluoroacetamido-39-deamino-3"-hydroxy-daunorubicin (VI) which? by treatment with aqueous sodium hydroxide, gives the compound (Ila) . Typically,, the N-tr if luoroacetyl group may be removed by mild alkaline hydrolysis, at a 10 temperature of 0°C by means of 0- IN aqueous sodium hydroxide. Glycoside (Ila) can be isolated as its hydrochloride by treatment v?it h hyd rogen chloride in methanol.

The 3v-deamino-4.9-deoxy-3 s~epi~4 5-epi~3 5 « 15 4'-epimino-daunorubicin, i.e. compound (la)r may be prepared by reacting 3'-epi-daunorubicin with salicylaldahyde so as to obtain the corresponding 35-epi-H-salicylidsne derivative; converting the 4 s-hydroxy group of the said 3'-epi-N-salicylidene derivative into a 20 trifluocomethanesulfonate group; and removing from the 35-epi-N-salicylidene-4 5~Q~tr ifluoromethanesulfonate thus obtained the salicylidene group by acid hydrolysis so as to cause the 3'-deamino-4"-deoxy-3'-epi-4'-epi-3", 4'-epimino-daunorubicin to be obtained via displacement of 25 the 4 ,J-0~tr if luoromethanesulfonate group„ The compound (la) may therefore be prepared by reaction of the 3'-amino group of 3'-epi-daunorubicin (III) [F. Arcamone„ A. Bargiotti, G. Cassinelli : Ge r,Patent 2752115 (June 1, 1978)) with sailcvlaldehyde , in a mixture of water and acetone at room temperature, to obtain the corresponding 3'-epi~N~s©licylidene derivative (IVc) which by treatment with t r i filuo r ome thane sulfonic anhydride in anhydrous methylene dichloride and in the presence of pyridine gives the corresponding 3'-epi-N-salicylidene-4?«0~t ri fluoromethansulfonate (ivd). This compound, dissolved in methanol£, can then be subjected to acidic hydrolysis of the salicylidene protecting group by means of p-toluensulfonic acid at room temperature to give, via the displacement cf trifluoromethanesulfonate leaving group, the desired compound of formula (la).

The compound (lib) can be prepared by bromination of (lis) followed by treatment of the resultant 14-bromo derivative with aqueous sodium formate at room temperature, according to the procedure described in United Patent Specification No. 3803124. It may be isolated as its hydrochloride in the same manner as glycoside (Ila).

The processes of the invention are summarised in the reaction scheme below.

The present invention also provides a pharmaceutical composition comprising as active ingredient ar. anthracycline glycoside of the invention or 5 pharmacr.^tically acceptable acid addition salt thereof together with a pharmaceutically acceptable carrier or diluent™ A therapeutically effective amount of a compound of formula (II) is combined with an inert carrier. 5 Conventional carriers may be used and the composition may be formulated in conventional manner. methods of treatment of the human or animal body by therapy. In particular, the compounds of the invention are 10 useful as antitumor agents- The compounds of the invention are useful in :och, 1 a III *» iv c — a 12 I V e VI c R„: OOHC-H.CH- 2 ©4 R3s OH d R2: OOHCgH4Cd~ R3: 0S02C?3 - 6 - e : COCFg The following Examples illustrate the invention.

EXAMPLE 1 3 3-epi-N-salicylidene-daunorubicin (IVc) 5 A solution of 2 g of 31-epi-daunorubicin (III) in a mixture of 80 ml of water and 20 ml of acetone,, was treated at room temperature with 0«5 ml of salicylaldehyde at ph 8« After 10 rains„ ethyl acetate was edded and the organic phase separated off£, washed with water twice,, dried 10 over anhydrous sodium sulphate, filtered and evaporated to dryness under vacuum.

The residue was first triturated with hexan<-' • o eliminate the traces of salicylaldeyde, then collected an:; dried under vacuum at 30°C to give (IVc) in almost 15 quantitative yield™ Rf 0.21 on TLC Kies^lcel F 254 (Merck) using as eluent the solvent mixture CH^Cl^-Acetone (B/2 v/v) .

EXAMPLE 2 3 *-deamino-4u-deoxy-35-epi-49-epi~ 3g, 4 9-epiminu-20 daunorubicin (la) To a solution of 2 g of 33-epi-N-salicylidene daunorubicin (IVc) in 20 ml of anhydrous dichloroinethane and 2.ml of dry pyridine kept at -10°C, was added a solution of 0-8 ml of trifluoromethane sulfonic anhydride 25 in 10 ml of dichloromethane. After 1 hour at -10°Ce, the mixture was diluted with dichloroinethane and washed with _ 7 - water, cold 0.1M hydrochloric acid, cold aqueous 5% w/v Sc^ hydrogen carbonate and water. The organic phase, dried over anhydrous sodium sulphate, was filtered off and the solvent removed in vacuo to give (IVd) Rf 0.50 on TC 5 Kieselgel F 254 (Herck) using as eluent the solvent fixture CH2C12-Acetone (95/5 v/v).

The crude product was dissolved in 50 ml of methanol and 0„2 g of p~toluensulfonic acid monohydrate was added. The solution was kept at room temperature for 1 10 hour, then 100 ml of water was added and extracted with little dichloroinethane. The aqueous phase was adjusted to PH 8 with 0.1M sodium hydroxyde and dichloromethane added. The organic phase was separated offr washed with waterv dried over anhydrous sodium sulphate and the solvent 15 evaporated to a small volume.

The mixture was purified by chromatography on a column of silica gel buffered at pH 7 using dichloroiaethane-ethanol as eluent. The eluate containing the product (la) was washed with water, evaporated in 20 vacuum, picked up with a little dichloromethane and crystallised FD MS 509 IH+J n.p. 135-137°C Rf 0.38 on TLC Kieselgel ? 254 (Merck) using as eluent the mixture Cn3 CI,-Cn3 OH~CHjCOOH-H2 0 (30/4/1/0.5 v/v) . 1»H-NMR (200 KHz, CDC13): 25 8.02 (dd, J-=1.1, 7.7Hz, 1H, H-l) 7.76 (dd, J-7.7, 7.7Hz, lHf H~2) „ 8 - 7. 37 (dd, J=l.l, 7.7Hz , IH, H-3) 5. 31 (dd e J = 3.0, 4.8Hz , IH t H-l5) 5. 17 (ddi J-2.0, 3.6Hz , IH, H-7) 4 . 32 (qd, J=<1, 6.7Hz , IH f H-5') 4. 07 (S, 3H, OCH3-4) 3. 17 (dd, J^l9,2Hz, IH , H-lOe) 2. 95 (dr J=19.2Hz, IHg H-lOax) 2 - 46 (666, J~2„ 02.0, 15.0Hz, IH, H-8e) 2. 43 (S, 3H, COCH3) 2. 30 (ddd, J=1.5, 4.3/ 6.4Hz, IH, H-3') 1.9-2.0 (mff 2Hi H-8ax, H-2uax) 1.87 (odd, J~1.5P 3.0, 14.6H2, 1H. H2se) 1.44 (df J=6.7Hz, 3H„ CH3~5') EXAMPLE 3 15 3'-deamino-4u -deoxy-?a-hydroxy-4'-epi-41-amino- daunorubicin (Ila) The title compound was prepared starting ti om the aziridine la. Ig of la was transformed into the K-tri fluoroacetyl derivative Ve by treatment with 1.2 ml of 20 trifluoroacetic anhydride in anhydrous methylene dichlorIds«, After work up the crude material [Rf 0.'/ on TLCe Kieselgel F 254 (Merck) using as eluent the solvent mixture CB^Cl^-Acetone (4/1 v/v)] was dissolved in 20 ml of acetone and treated with a catalytic amount of sulfuric 25 said at 10°C. - 9 - The mixture was diluted with 200 ml of methylene dichlor ide, washed with water? aqueous 5% w/v soc. . hydrogen carbonate and water. The solvent was removed in -i vacuum and the residue purified on a column of silica gel 5 using methylene dichloride as the ©luting system to afford 0.7 g of pure VI.

Rf 0.21 on TLC, Kieselgel F 254 (Merck) using as eluent the solvent mixture CHaCI2-Acetone (4/1 v/v).

The product VI u«as slowly dissolved in aqueous 10 0.IN sodium hydroxide, at 0°C in order to perform the hydrolysis of the N-trifluoroacetyl protecting group.

After 1 hour at 0°C, the solution was adjusted to pH 8.6 with 0.IN hydrochloric acid and extracted with methylene dichloride. The solvent was evaporated of £, 15 affording 0.5 g of a residue that was converted by treatment with methanolic hydrogen chloride into the hydrochloride of 4'-deoxy-4'-amino-4'-epi-3'-deamino-3'-hydroxy-dounorubi cin.

MS FD 527 {H+j( a.p.153°C (dec). 20 Rf 0.18 on TLC Kieselgel F 254 (Merck) using the solvent system CH2 Cl2-CH3 OK~CH3 COOH~H2 O (30/4/1/0.5 v/v) 1«H - NMR (200 MH„CDC1-) «& 8.02 (dd, J«0.9, 8.5Hzf IH, H-l) 7.77 (dd, J~8.5, 8.5Hz, lHf H-2) 25 7.38 (dd, J-0.9, 8.5Hz, IH, H-3) 5.52 (dd, J«=<1, 4.0Hz, IH, H-l' ) - 10 - 5. 28 (dd, J=l.8, 4.0Hz, IH, H-7) 4.07 (s, 3H„ OCH3-4) 3.69 (dq, J=6.3, 9.5Hz, IH, H-5') 3.51 (ddd, J=48, 9.5, 11.6Hz, IH, H-33 ) 5 3.22 (dd, J=1.9, 18.9Hz, IH, H-lOe) 2.94 (d, J-18-9Hz, IH, H-lOax) 2.40 (si? 3He, COCH3) 2.2-2.4 (m, IH, H-8ax) 2.30 (dd, J=9.5, 9.5Hz.. IH, H-49) 10 2.0-2.2 (rn, 2H, H-8e, H-2 'e) 1.70 ' (ddd, J = 4.0, 4.6, 13.2Hz, IH, H-2"ax) 1.31 (d, J=6.3Hz, 3H, CH3~53) EXAMPLE 4 3 v»deamino-38-hydroxy-4 i-dsoxy-4 ,J-epi~ 4 * -15 amino-doxorubicin (lib) 0 „ 5 g of Ila was dissolved in a mixture of methanol and dioxane„ The solution was treated,, as described in United Patent Specification No.3,803,124 first with bromine to give the 14-bromo-derivative and then with 20 aqueous sodium formate to give the title compound.

This was converted into its hydrochloride by treatment with methanolic hydrogen chloride. FD-MS 543 [M+ J t, TLC on Kieselgel F 254 (Merck) using the solvent system CH2C12-CH30H-CH3C00H~H,0 (30/4/1/0.5 v/v) Rf 0.10. - 11 - The cytotoxic activity of the new anthracycline glycoside of the invention (Ila) was tested 8,in vitro" against HeLa cells P388, P388/DX, LoVo and LoVo/DX.

Time of exposure to the compound: 24 hours/in 5 comparison with daunorubicin.

The results are shown in Table 1.

The compound, when tested "in vitro" against P-388 ascitic leukemia and Gross leukemia, exhibited good antitumor activity, in comparison with daunorubicin, 10 especially when orally administered.

The results are given in Tables 2 and 3.

Table 1 In vitro activity of 3deamino-^'-deoxy-31 -hydroxy-^1 -epi. =4'-amino-aaunorubicin (Ila) in comparison with DMR ID 0(ng/ml) Compound _ „ _ „ b) c) d) e) f) HeLa P363 P388/DX LoVo LoVo/Dx DMR . 19 10,5 730 43 820 111 24.5 235 37 230 a) Dose giving 50% reduction of cell nuabor in comparison with untreated controls, b) Human cervix epithelioid carcinoma cells c) P 388 leukemia cells sensitive to Doxorubicin . . d) P 3S8 leukemia cells resistant' to Doxorubicin e) Hunan colon adenocarcinoma cells sensitive to Doxorubicin f) Human colon adenocarcinoma eel 1.3 rejistar.t to P^.. a ubicin _ 1 3 _ a Table 2 Effect against p 388 ascitic leukemia ^ ^ Compound dose T/C% Toxic deaths DIJH 2.9 155 0/10 kA 170 8/10 Ila 1»96 155 0/10 2-9 150 0/10 ' 140 9/10 6-6 100 10/10 Evaluated on tha basis of autoptic findings. - 1 (I - 3 Table 3 Effect against Gross leukemia Compound doseb T/C%° mg/Kg DNR 10 165 C/20 15 192 2/20 Ila 8.2 . 175 0/20 11.5 230 0/10 is: i 240 0/10 22., 5 130 0/10 aLxpegiments ye re performed in C3H mice. inoculated with 2x10 leukemia cells i.v. fo Treatment i.v. vu day 1 after tumor inoculum. c Median survival time of treated mice/median survival tine of controls x 103. d Evaluated on the basis of autoptic findings. « . d J.OX1C aeatns

Claims

- 15 -CLAIMS

1. An anthracycline glycoside having the genera: formula (II): ft oh COCHjR, :h,6 6 in (id where j ^ represents a hydrogen atom or a hydroxy! group, 5 and phartncceutically acceptable acid addition salts •.hereof.

2. - ft compound according to claim 1 f which is 3 '-deanino~4 u—3eoxy-3 '-hydroxy-4 ! -epi-4 e-amino-dauno rubicir, or its hydrochloride* 10 3. A compound according to claim 1, which is

3. S-deamino-4'-deoxy-3"-hydroxy-4'-epi-4 5-amino-doxorubicin or its hydrochloride.

4. A process for the preparation of a glycoside of formula (II) as defined an claim 1 or a pharmaceutically 15 acceptable salt thereof, which process comprises converting 3'-deamino-4*-deoxy-3'-epi~4'-epi-3®,4'-epimino--daunorubicin into the corresponding N-trifluoroacetyl derivative? converting the said N-trifluoroacetyl derivative into 4 a-deoxy-4"epi-tritluoroacetamid o~ 35-deamino-39-hydroxy daunorubicin; removing the N-trifluoroacetyl group from the 4'-deoxy-4'-epi-fcr if luoroacetamido~3 v-deamino-3 ® -hydroxy 5 daunorubicin so as to obtain the glycoside of formula (II) wherein is a hydrogen atom? if desired, converting the said glycoside of formula (II) into a pharmaceutically acceptable acid addition salt thereof? if desired* brominating the said glycoside of formula (II) or 10 pharmaceutically acceptable salt thereof and hydrolysing the 14-bromo derivative thus obtained so as to form the glycoside of formula (II) wherein R^ is a hydroxy group; andt if desired, converting the said glycoside of formula (II) wherein R^ is a hydroxy group into a pharmaceutically 15 acceptable acid addition salt thereof,,

5. A process according to claim 4, wherein the 3 a"deamino-4 9-deoxy-39-epi-4'-epi-3',4'-epimino-daunorubicin is reacted with trifluoroacetic anhydride to obtain the corresponding N-tr ifluoroacetyl derivative 20 which, by treatment with a catalytic amount of sulfuric acid in acetone gives 45-deoxy-49epi-trifluoroacetamido-3s-deamino -3'-hydroxy-daunorubicin; removing the N-trifluoroacetyl protecting group therefrom bv mild alkaline hydrolysis,, at a temperature of 0°C, by means of 25 0.IN ageuous sodium hydroxide? optionallyf isolating the glycoside of formula (II) wherein is a hydrogen atom as its hydrochloride by treatnent with hydrogen chloride in methanol? optionally, converting the said glycoside of formula (II) or hydrochloride thereof to the glycoside of formula (II) wherein R1 is a hydroxy group by* bromination followed by treatment of the resultant 14-bromo derivative with aqueous sodium formate at room temperature? and,-optionally,, isolating the said glycoside of formula (II) wherein is a hydroxy group as its hydrochloride by treatment with a methanolic solution of hydrogen chloride.

6. - A process according to claim 4 or 5 wherein the 3'-deamino-4°-deoxy-35-epi-4"-epi-3, 4'-epimino-daunorubicin is prepared by reacting 3!-epi-daunorubicin uith salicylaldehyde so as. to obtain the corresponding 3 -epi-N-salicy 1 ioen'i- derivative? converting the 4'-hydrcxy croup of the said 3 * - epi -N-salicvlidene derivative into a trifluoromethanesulfonate group? and removing from the 3 * epi-N-salicylidene-4'-0-tr ifIuoromethanesulfonate thus obtained the salicylidene group by acid hydrolysis so as tc cause the 3 '-deanino-4 '-deoxy -3 8-epi-4 8~epi«3'31 4'-epimino-daunorubicin to be obtained via displacement of the 41-0"tri£1uor omethanesulfonate group.

7. - A process according to claim 6, wherein 3'-epi~daunorubicin dissolved in a mixture of water and acetone is reacted, at room temperature, with salicylaldehyde to obtain the corresponding 3*-epi~N~salicylidene derivative which is subsequently - 18 - treated, in anhydrous methylerv: dic'ulor ido ar-.i in presence of dry pyridine? with tr if 1 uoromethanesul f or. ic anhydride to give the corresponding 'N-sa 1i cy1idene-3'--epi-4 "-O-trifluoromethanesulfonate of which the 5 salicylidene. protecting group is subjected to acidic hydrolysis by means of p-toluensulfonic acidf at room temperature with the said trifluorornethanesulfonate being dissolved in methanol, to obtain, via the displacement of the trifluoromethanesulfonate leaving group,, the 10 3 '--deamino-4 s-deoxy-3 "-epi-4'-epi-3 ' , 4 '-epimino-daunorubicin.

8. A pharmaceutical composition comprising an antnracycline glycoside of formula (II) as defined in claim 1 or a pharmaceutically acceptable acid addition 15 salt thereot- together with a pharmaceutically acceptai ie carrier or diluent.

9. ?. An anthracycline glycoside of formula (IX) as defined in claim 1, or a pharmaceutically acceptable salt thereof, for use in a method of treatment 20 of the human or animal body by therapy.

10. An anthracycline glycoside or salt thereof according to claim 9 for use an an antitumor agent,

11. A process for the preparation of an anthracycline glycoside of formula (II) as defined in claisr. 25 1 or a pharmaceutically acceptable salt thereof* said process being substantially as hereinbefore described in (' - 19 - Example 3 or Examples 3 and 4 together.,

12. An anthracycline glycoside, as defined in acid addition claim 1 or a pharmaceutically acceptable/salt thereof whenever prepared 5 by a process as claimed in any of claims 4 to 7 or claim 11.

13. „ An anthracycline glycoside _ as defined in claim 1 or a pharmaceuifeically accept!!^e/salt thereof substantially as hereinbefore described with reference to Example 3 or Examples 3 and 4 together, 10 as defined in Claim 8,

14. A pharmaceutical composition/substantially as hereinbefore described by way of Example. Dated this 10th day of June? 1987 BY: TOMKINS & CO™, ^ (Signed) Applicants' Agents, 5 Dartmouth Road,, Dublin 6. 20 25 30 35