CA1184903A - Process for preparing 3'-deamino analogs of glycoside anthracycline antibiotics - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE

Antitumor anthracycline glycosides of the formula:

wherein R1 is hydrogen or hydroxyl and R2 is the .alpha.-L-pyranosyl-hexose II or III

II III

are prepared from 3',4'-diepi-analogs of daunorubicin and doxo-rubicin by deamination in a cold acidic medium, such as 1N aqueous acetic and at 0°C with NaNO2. In this series of new anthracyclines showing antileukemic activity against murine leukemias, the doxo-rubicin analogs, when parenterally administered display an activity comparable or superior to that of doxorubicin and they are also active orally against Gross murine leukemia.

Description

1 The invention relates to a new class of glycosidic anthracyclîn~ antibiotics, which are related to daunorubicin and doxorubicin, their preparation, pharmaceutical compositions con-taining them and the use thereof in treatin~ certain mammalian turnors.
Daunorubicin and doxorubicin~ to which the compounds of the present invention are related~ are both known and used anti-tumor antibiotics and both are amply described in the literature.
Moreover, the starting material for the present compounds: 3',4'~
1~ diepi-daunoru~icin. and 3',4'-diepi~doxorubicin are also bo~h known compounds described in U.S. Patent 4,112,076 ~Sept, 5th, 19781 which .is ownecl by the unrecorded assigne~ hereof, The invention provides, in one aspect thereof, a new cL~ss of anthracycline glycosides of the formula I

O OH
~ ~ ~ ~OCH 2 Rl El CO O OH

~here.in Rl is hydrogen or h~droxy and R2 is the ~ pyranosyl residue of the formula II or III

,~
H C - - - r~ ~ HC~ f III
O~ OH

These anthracycline glyco~ides contain a neutral glycoside moiety, 1 either 2,3,6-trideoxy~ glycero-hexopyranosid~ ulosyl (u-I,-cinerulosyl, II) or 2,6-dideoxy- u.-L-arabino-hexop~ranos~ [~), and are named as Eollows:
(1) 3'-deamino~4l-dehydro-claunorubicin (I: Rl--H; R2=II) (h~rein-after compound I-A~;
~2) 3'-deamino-3'-hydroxy-4'-epi-dauno:rub;icln (I: ~ l, R2-III) (hereinafter compound I-B);
~3l 3'-deamino-A'~dehydro-doxc)rub:ic.irl (:t: Rl--O~I~ E~2-~I.t~ (here.in-after compound I~C); and C4~ 3'-deamino-3'-hydroxy-4'-epi-doxorub1cin (I: Rl-~0El, R2=III) (hereinafter compound I-D~, Ir. another aspect, the invention provides a process for prepariny the anthracycline glycosides of the invention, The new compounds are prepared from 3',4'-diepi-daunoru~icin (IV, Rl=
Hl and 3',4'-diepi-doxorubicin (IV, Rl-0H), both known anthra-cycline glycosides described in the aforesaid U.S. Patent 4,112,076 (,Sept. 5th, 1978~. 0 OH

~ 0CH2R
H3CO o OH `~

I IV

HO.

the process for the preparation of the new compounds comprises deaminating the anthracycline glycosides of the formula IV~ in which an axial amino group neighbors an equatorial hydroxy group, proceedi,ng via a diazotized intermediate to CJiVe compouncls I-A

2~

and I-C as the result o~ hydride migr~tion from the C-~' position and compounds I-B and I-D by simple nucleophilic attack of water at the C-3' position.
The deamination reaction is pref~rably performed ~ith sodium nitrite in a cold aqueous acid rnedlum, hut it m~y alterna-tively be carried out with nitro~en -trloxide (~123) ~ alkyl nitri.tes in cold aqueous or alcoholic m~cli~.
The new anthracycline cJlyco~i~e~ of th~ irlv~nt:ion d.i~
play antitumor activity. Part:ic~lAlarl.y, 3'~deam.irl~-3'-hydroxy~
1~ epi-doxorubicin ~I-D~ displays remarkable act.iv.ity on experimental tumor~ in mice. Accordingly, the invention, in yet further aspects thereof, provides pharmaceutical compositions comprising a thera-peutically effective amount of an anthracycline glycoside of the fo.rmula I admixture with a pharmaceutically acceptable diluent or carrier, and methods of treating certain mammalian tumors com-prising administering to a host afflicted therewith, therapeutical-ly effective amounts of said compounds, The invention will now be described in greater detail in the following preparative examples and biological data, ... . _ ..

3'-deamino 4'-dehydro-daunorubicin (I-A) and 3'-deamino-3'-h~droxy-

4'~epL-daunorubicin ~I~Bl.
A solution of 0,55 g; 1 mmol oE 3'-4'-diepi-daunQrubicin hydrochloride (IV: R~ in 25 ml oE water was cooled to 0C and then treated wit~ 0.72 g; 7,5 mmol of sodium nitrite and 7.5 ml - of 1 N aqueous acetic acid in several portions over a period of 20 minute5 under stirring at a rate such that the temperature did not exceed 0C. After 3 hours at 0C, the reaction mLxture, con-tain~ng a red colored precipitate, was brought to room temperature.
Nitrogen was bubbled through -the solution -to remove excess nitrous 1 acid, and the solution was then ~xtract~d with chloroform. The chloroform extract was washed with wat~r, dried ov~r anhydrous sodium sulphate, evaporated to a smaLl volume and chromatoc3raphed on a silica gel column. Elution of the column with chloro~orm afforded 3'-deamino-4'-dehydro-daunorubicin (I-A~ a~ an amorphous solid havin~ a m.p. oE 143-1~4C (wlt:h d~cornposition); [~23 +190 ~c=0.05, in methanol). U.V. ~nd VCS ~p~-ckrcl: ~ CE3011 235, m~x 253, 290, 480, 4~6 and 530 n~ E~Cnl 808, 557, 1~, 236, 2~2 arld 143). Field desorption m~s~ spectrum: m/~ 510 ~M~'); 398 and 562.
H-NMR spectrum (CDC13~: 1.36 (d, ~=6.5 Hz, 3El, CEI3-S ~; 1.70 -2.40 ~m, 6H, H-8, H-2', H-3'); 2.41 ~s, 3H, COCEl31; 2.97 ~d, ~~
19,5 Hz, lH, H-lOaX~; 3.25 (dd, J=19~5 Hz, lH, H-10 qli 4.09 (s, 3E~ OCH ); 4.40 ~q, J=6.5 Hz, lH, H-5'~; 4.62 (s; lH, OH-9~; 5.41 ~m, lH, H-71; 5.67 (t, J=6.0 Hz, lH, H-l'); 7.30-8.10 ~m, 3H, aromatic protons~; 13.2~, 14.01 ~ (two s, 2H, OH-6, OH-ll).
.
Further elution of the column with a 97:3 chloroform:
acetone mixture gave 3'-deamino-3'-hydroxy-4'-epi-daunorubicin (I-B~ haYing a m.p. of 165-170C ~with decomposition), ~a]D
-~350 (c=0.025, in methanoll, U.V. and VIS spectra: ~ CaH3H 235, ao 254, 2~Q, 480, 4~6 and 530 nm ~El%Cm = 714, 509, 154, 226, 231 and 142~, Field desorption mass spectrum m/z 528 ~M 'I; 398, 362, H-NMR spectrum (CDC13~: 1.36 (d, J=6.0 Hz, 3H, CH3-5'~; 1.60-2.40 ~m, 4H, H-8, H-2'~; 2.41 (s, 3H, COCH3); 2.86 and 3.30 ~t~o d, J=l9 ~z, 2~, H-10); 3,5-4.0 (m, 2EI, H-3', H 4'1; 4.07 ~s, 3H, OCH3~; 4.65 ~m, lH~ H-5'~; 5.25 ~m, lH, H~71; 5.48 (m, lH, H~
7.30~3.10 (m, 3H, aromatic protons), 13.26, 13.98 ~ ~two s, 2H, OH~6, OH~
EX~MPLE 2 3'~deamino-4'-dehydro-doxorubicin_(I-C) and 3'-deamino-3'-ll~droxy 4'-epi-doxorubicin ~I-Dl~
~ . .

1 The preparation of the title compounds (I-C) and (I-D) starting from 3',4'-diepi-doxorubicill (IV: Rl=O~I~ was ~erformed according to -the procedure described in ~xampl~ 1.
3'-deamino-4'-dehydro-cloxorubicin (I-C) was obta.ined as a red colored powder havincJ a m.p. oE 3~0VC (~.Jith decompositisrl), [~]D = ~ 5 (c=0~046, in methanol). U.V, and VI~ r;pectr;l A C~l30EI
~ c mc.li~
~35, 254, 290, 480, ~96 an~l 530 nm ~E'j~ 6~0, ~'~0, ~ O, 2~, 23~, 140). Field desorption ma3~; ~E~ectra: m/~, ~2~) ~M~ , 33~.
C-NMR spect.rum (CDC13~ .9 (C-6'); 27.6 (C-2'); 33,1 (C-3');
33.9 ~C-10); 35.9 (C-8~; 56,6 (0-CE[3); 65~ C~ ; 68.9 (C-7~;
71.3 C-5'l; 76.6 (C-9); 100.3 (C-l'); 11~.5 ~C-3); 119,8 ~C-l, C-4a~, 133.7(C-6a, C-lOa, C-12a); 135.8 (C-2)i 155.6(C~ ; 156.1 ~C-6~, 161,0 (C-4); 186.9 (C-5, C-121; 210.3 ~C-4'1 and 213.9 .C-13l.
Similarly, 3'-Deamino-3'-hydroxy-4'-epi-doxorubicin CI~D) was obtained as a red colored powder having a m.p. of 176~180C
(with decompositionl; [~323 = ~280 ~.c+0.046, in methanol). U.V.
and VIS spectra: ~ ma3 235,254, 290, 480, 496 and 532 nm ~El%Cm = 674, 478, 186, 223C 228 and 14810 Field despor-tion mass spectrum: m/z 544 (M ~, 41~, 378, 336. 13C-NMR spectrum (CD30D:
CDCl3, 1:1 by volume3: 17.8 (C-6'); 33.9 (C-10); 36.3 ~C-8); 3,80 (C-2'); 57.0 (OCH31; 65.4 (C-14); 68.8 (.C-7); 69~8 (C-3'~ C-4'~;
77.~ (C-~ 77.9 (C-5'1; lQl.4 (C~ ; 119.3 (C-3, C-4a~; 120.3 ~C-ll; 134.3 and 135.8 (C-6a, C-lOa) C-12a); 136.5 (C-2~; 155.7 ~C-6, C-11~; 161.6 (C-41; 187~3 (C 12).i lB7.6 (C-5) and 214.2 13~.
BIOLOGICAL AClIVI~Y
The cytotoxic activity of the new anthracycline glyc~-sides of the invention were tested a~ainst He I,a cells "in ~itro"
(time of exposure to the compoun~s: 24 hours/in ~omparison t~itE~

1 daunorubicin and cloxoru~icin. The r~sults ar~ shown in Table 1.

- T~ble 1 - Colony inhibition t~st ~cJainst ~lel,cl cells "in vi.tro"

__~__ Compound ID50 (ncJ/mL) Daunorubicin.HCl .l2 Compound I-~ ~50 Compound I-B 60 Doxorubici.n.EICl 1l.
Compound I-C 40 Compound I-D 25 Certain of thecompoundsof this inventionr when tested against P-388 and L 1210 murine leukemias exhibited supPrior results in comparison with daunorubicin and doxorubicin, especially when orally administered~ The results of the antitumor activity against different murine leukemias are given in Table 2, 3 and 4 below~
Table 2. Antitumor activity against P388 leukemia.
Treatment ip on Day 1.

2~ .
Compound Dose T/ca Tsb Toxic (mg/kg) % L deathSc Daunorubicind 2.9 170,150 0/9,0/10 nfs, o/lo 4.4 154,150,155 0~8,0~9,0~10 2/~,3/9,Q/10 6.6 145,150,160 0/8,0/9,0/10 7/8,5/9,0/10 Compound I-A 30.8 190 0/8 0/8 127 0~'8 0/8 160,165 0/3,0J10 0,~3,2/10 78 95 0~10 ~/lQ
101 100 1/10 5flO

3~

?~ 3 'rabl~ 1 cont.

Compound (mg/k~ /ca L,Tsb Toxi.c c deaths Doxorubicine 4.4 170 0/lO 0/10 6.6 .160 0/lO ~,/lO
Compound I-Be 15 :L80 0/'i 0/~
290 0/~ 0/5 26 16() 0/S 3/5 Doxoru~icine 4.4 225 0/8 0/8 6.6 290 l/8 0/~

Compound I-Ce lO 170 2/lO 0/lO

22,5 21~ 0/~ 0/8 33.7 230 l/8 0/8 Doxorubicin 4.4 245 r 223 0/lO,0/lO 0/lO,Q/lO
6.6 255,258 l/lO,0/lO 0/lO r 0/10 ~0 34~,282 2/l~,l/lO 0/lOjO/lO
Compound I-D 6,6 250 l/lO 0/lO
2Q lO 2~0 3/lO 0/lO
>630,>658 6/lO~6/lO ~ lO
22~5 >Ç30,>658 ~/lO,5~1Q 0/lO,2/lO
33.~ 194 3/lO 5/lO
50,5 76 l/lO 7/lO

aMPdian survival time; % over untrea-ted controls Long term surYivors ~3 60 days) CEvaluated on the basis of autoptic findings on dead mice dData of three experiments Dissolved in 10% Tween 80 Dissolved or suspended in H20; data oE two experiments *Trade Mark -7-u to tn o o o o o o o o U ~ r,o r-l r-l 1-- t~ tx) r~ ,, r~l r-l r-l r~l 1.O ~ tO r~ n "
X (~ O rl ~D O O t~) O O O C ) O C~\ O O t~l O O t~ V O O
O (I) E-l r~ ~

U~ tiO r,~ O O O O O t~> t~) O
E-l t ~ ~ r~l t~ CO t~O 1~ r-l r-¦ r-¦ r~l r~l ~9 t~ ~ r~l r I r~-l C;- t.~ (~
td o o ~1 o o o o o o o o t~ ~ o o o o o o ~ O
X ~,''`1O ~1 rn Ln n ~ n o ~ ct~ co ~o ~o r~ a In c~ r~ co ~n In ~r o o ~ ~ I` r~ co ~ Ln ~ Ln Ln ~o ~o 1~ o o ~ In r~ o o ~1 1 0 O O~1 r~ r-l~ ~ ~I r~l ~I r~~ r~¦ r-l ~0 Ln E~ ~ o\ O~ ~ A O
E~ ~ ~1 ~7 co t~ ~
td . a) t~ t~ t~u:~ cn cs~ r ~ ro ~ t.~ t~ a) a),Y . . . -a n ~o ~ 0 t~ ~D t~ ~r ~r ~o t~ t ~ o o ~ ~D t~ o o O ~ r-l r-l r~lr-lt~ ~)~ t~) ~r r~l r1 ~ r~l ~I r~l r1 C~ tr)~, a ~
r~
o ,~ a a s~ H
O S~ U U ~.) ~
o oQ~ ~ ~
S-l O O h O ~1 0 ~3 O ~ X E; ~ X ~ X 1~ h u~ O O O O O O O e ~s V ~ X
~ a .~ ,~ a~ Q) a) o O ~ O
r~ ~ a) ~ a~ 3 ~;!0 ~ ~ 1: 3 . 3 ~ ~
~ O

,~ ~ a ~ ,, ,_~ ~
h t'~
a) ,~
.
~ ,1 h Q-p; . O r~ rl N
a) .Q
~ ~ E~
pO ~rl r~ ~rl ~rl 8 3~ u~
t-h U~ o ~
~0 O ,~ ~
E~ ~ ~1 ~

*Trade Mark '303 1 Table 4. Effect of compound I-C agalnst Gross l.eukemia Compound Vehicle Route Dose T/C LTsb Tox c c Doxorubicin Tween 80 iv 13~ 21~,275,2~2 0/20 0/20 16.9e 228,175,250 1/2B 8/2~
Compound I-C " iv ~0 185,250 0/17 2/17 0/J.0 7/~.0 57.~ .ll6 0/10 10/10 Compound I-C " oral ~0 192 0/~ 0/~
48 200 0/~ 0/~
1~ 57.6 200 ~/8 2/~
. _ . ~ . . .. . . . . .
a~D c see Table 2 dSee Ta~le 3 Data of three Experiments Data of two experiments Variations and modifications can, of course, be made without departing from the spirit and scope of the invention~
Haviny thus described our invention, what we desire to secure ~y Letters Patent and hereby claim is:

Claims (10)

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A process for preparing an anthracycline glycoside of the general formula I:

I

wherein R1 is hydrogen or hydroxy and R2 is:

II III

characterized in that an aqueous solution of the known 3',4'-diepi-daunorubicin hydrochloride or 3',4'-diepi-doxorubicin hydrochloride of the general formula IV:


1. Claim 1 continued...
   
   wherein R1 is hydrogen or a hydroxy group, is separately re-acted at 0°C and for 3-4 hours with sodium nitrite and 1N
   aqueous acetic acid to obtain, respectively, a deaminated mixture of the glycoside of formula I (R1=H; R2=II and III) or of the glycoside of formula I (R1=OH; R2=II and III), submitting each mixture to a chromatographic separation on a silica gel column by first eluting with chloroform and when with a chloroform-acetone mixture 97:3 (v/v), whereby each said compound of formula I is recovered as free base in pure form.
2. 2. A compound of the general formula I as defined in claim 1 whenever prepared by a process as claimed in claim 1 or an obvious chemical equivalent thereof.
3. 3. A process as claimed in claim 1 for preparing 3'-deamino-4'-dehydro-daunorubicin which comprises reacting 3',4'-diepi-daunorubicin hydrochloride with sodium nitrite and acetic acid and separating and recovering the subject compound.
4. 4. 3'-Deamino-4'-dehydro-daunorubicin whenever prepared by a-process as claimed in claim 3 or an obvious chemical equivalent thereof.
5. 5. A process as claimed in claim 1 for preparing 3'-deamino-3'-hydroxy-4'-epi-daunorubicin which comprises reacting 3',4'-diepi-daunorubicin hydrochloride with sodium nitrite and acetic acid and separating and recovering the subject compound.
6. 6. 3'-Deamino-3'-hydroxy-4'-epi-daunorubicin whenever prepared by a process as claimed in claim 5 or an obvious chemical equivalent thereof.
7. 7. A process as claimed in claim 1 for preparing 3'-deamino-4'-dehydro-doxorubicin which comprises reacting 3',4'-diepi-doxorubicin with sodium nitrite and acetic acid and separating and recovering the subject compound.
8. 8. 3'-Deamino-4'-dehydro-doxorubicin whenever prepared by a process as claimed in claim 7 or an obvious chemical equivalent thereof.
9. 9. A process as claimed in claim 1 for preparing 3'-deamino-3'-hydroxy-4'-epi-doxorubicin which comprises reacting 3',4'-diepi-doxorubicin with sodium nitrite and acetic acid and separating and recovering the subject compound.
10. 10. 3'-Deamino-3'-hydroxy-4'-epi-doxorubicin whenever prepared by a process as claimed in claim 9 or an obvious chemical equivalent thereof.