DE3428945A1 - 4'-HALOGENANTHRACYCLINGLYCOSIDES, METHOD FOR THE PRODUCTION THEREOF AND THE PHARMACEUTICAL PREPARATION CONTAINING THEM - Google Patents

Description

FARMITALIA CARLO ERBA S.P.A. IN MILAN (ITALY)

4th ' -Halogenanthracycline glycosides, processes for their preparation and pharmaceuticals containing them To ₇

preparation

This invention relates to 4'-haloanthracycline glycosides, to a process for their preparation and to pharmaceutical compositions containing them.

The invention thus provides anthracycline glycosides of general Formula (A)

0 OK

COCK ₂ X

wherein X is a hydrogen atom or a hydroxy group, R _{1 is} a hydrogen atom or a hydroxy or methoxy group and R _{2 is} a bromine, chlorine or fluorine atom. ^ Pharmaceutically acceptable acid addition salts of these anthracycline glycosides are also encompassed by the invention.

The compounds of the invention in which X is a hydrogen atom can by an exchange reaction between a compound of the general formula (II) in which R _{1 has} the above meaning and R _{3 is} an amino protecting group, and a fluoride, chloride or bromide and subsequent removal

wUi-f

- ο .. -

of the protecting group from the resulting compound of the formula :-( III) ". This procedure is followed by the reaction slip below explained. The compounds of the invention "wherein X represents a hydroxy group can be selected from the corresponding Compounds of the invention wherein X is hydrogen> by bromination and subsequent treatment of the resulting 14-bromo derivative with aqueous sodium formate according to the process described in US Pat. No. 3,803,124. "These processes are within the scope of the present invention.

£ PG CCi / C-'fj

COCH.

OH

(ID

CF ₃ SO ₂

COCH.

OH

(III)

A (X = H)

copy p: $

If R _{2 represents} a bromine or chlorine atom, the exchange reaction preferably takes place between the compound (ii) and tetra- (η-butyl) ammonium bromide or. -Chloride and the amine protecting group R ₃ is preferably a trifluoroacetyl group.

The reaction to remove the protecting group can then be a mild alkaline hydrolysis, for example with 0.1N aqueous sodium hydroxide solution. If R _{2 is} a fluorine atom, the exchange reaction takes place-preferably between the compound (II) and cesium fluoride and the amine protecting group R- is preferably a trichloroethoxycarbonyl group. The reaction to remove the protecting group can then be a hydrogenation, for example with zinc dust and acetic acid. The exchange reaction can be carried out in an organic solvent such as methylene dichloride or acetonitrile.

The starting materials (II) are either known compounds, for example 4'-epi-4'-0-trifluoromethanesulfonyl-N-trifluoroacetyldaunorubicin (II: R ₁ = OCH ₃ , R ₃ = COCF ₃ , see EP patent application 8117618.1) or are derived from known 4'-Epidaunorubicinderivaten easily prepared by reaction with trifluoromethanesulfonic anhydride.

The compounds according to the invention and their pharmaceutically acceptable acid addition salts are useful as anti-tumor agents. These new compounds of formula (A) are prepared in the same manner as the parent compounds (i.e. Daunorubicin and doxorubicin) for treating various experimental mouse tumors, e.g. L1210, P388, P388 / 04 and the like. used. Previous screening tests have found these new compounds to be more active in some cases are as the root links. Accordingly, the invention further provides pharmaceutical compositions comprising an anthracycline glycoside according to the invention or a pharmaceutically acceptable acid addition salt thereof in admixture with one pharmaceutically acceptable diluents or carriers.

EPO COPY / "

Biological activity tests

The compounds der_3ejLspiele 1, 2 and 3 were in vitro in comparison with daunorubicin (DNR) and doxorubicin - (DX) against HeLa cells and on DX sensitive and resistant P388 cells

5- tests (P388 / DX). The results are given in Table I.

All of the new derivatives appear to be more cytotoxic than their parent compound to HeLa and DX sensitive P388 cells. This increased cytotoxicity is, however, much more pronounced when one considers the activity of these compounds against P388 / DX considered. Here there can be a 100 to 250-fold increase in the cytotoxicity of these derivatives compared with the parent drug to be established. In vivo, these compounds were tested against three different experimental leukemias. ■ '' The antitumor activity obtained against P388 ascitic leukemia is given in Table II. The activity shown by the compounds of Example 3 is equal to that exerted by DNR, while the other DNR derivative, the compound of Example 1, has anti-tumor efficacy that is definitely better than those of DNR.

The compound of Example 2 has approximately the same activity as DX in the maximum tolerated dose (4.15 mg / kg). All new compounds show activity against P388 / DX leukemia, as indicated in Table III, while DNR and DX have no effectiveness. The three new links were too after IV administration tested against common gross leukemia, the results are given in Table IV. In this system, the compound of Example 3 is as active as DNR, while the compound of Example 1 appears to be more active than the parent compound. The compound of example 2 shows about the same effectiveness as DX. Two of these compounds were also tested after oral administration, wherein they showed interesting activity, while DNR and DX are inactive when administered orally.

ΐΡΥ f-:: '

3A28945

Table I.

Cytotoxic activity of the compounds of Examples 1,

and

link ID ₅₀ (ng / ml) HeLa ⁺ '- P388 ⁺⁺ P388 / DX ⁺⁺ DNR
example 1
Example 3
DX
Example 2 12.3
6, B.
5.8
12.5
3 2.8
0.5
2.4
4.25
0.2 980
4th
8.8
1250
7th

+ Colony inhibition test after 24 h exposure are the drugs

++ Cytotoxicity assessed after 48 h exposure to the drugs

Data from multiple attempts

- 11 -

Tabel

II

Activity against P388 ascitic leukemia

a)

link Dose ^b)
(mg / kg) T / C ^C)
% LTS ^d) toxic
Death.
cases ' DNR 2.9
4.4
6.6 165 155
170 135
130 115 0/20
0/18
0/20 0/20
0/18
12/20 example 1 2.9
4.4
6.6
10 145
160
195
280 0/10
0/10
0/10
3/10 0/10
0/10
0/10
0/10 Example 3 2.9
4.4
6.6
10 130
155
145
85 0/10
0/10
0/10
0/8 - ^ 0/10
0/10
0/10
6/8 DX 4.4
6.6
10 215
235
335 0/10
0/10
4/10 0/10
0/10
0/10 Example 2 2.4
2.88
3.46
4.15 200
230
210
325 0/10
0/10
0/10
4/10 0/10
0/10
0/10
1/10

a) Experiments were carried out on CDF mice, i.p. inoculated with leukemia cells

b) treatment i.p. on day 1 after tumor inoculation

c) Mean survival time of the treated mice / mean Survival time of controls χ

d) Long survivors (> 60 days)

e) Evaluated on the basis of findings at autopsy.

EPO copy

Tabel

III

Activity against P388 / DX ascitic leukemia

link dose
(mg / kg) T / C ^C)
% toxic
Death ,,
cases ^d) DNR 4/4
6.6 91
87 0/10
0/10 example 1 - _4t4 _-
10 143
143 0/10
0/10 Example 3 6.6
10 148
122 0/10
3/10 DX 6.6
10 108
117 1/20
1/20 Example 2 2.88
3.46
- 4.15 122
125
137 0/20
0/20
1/20

a) Experiments were carried out on BDF. mice, i.p. inoculated with 10 cells

b) treatment i.p. on day 1 after tumor inoculation

c) Mean survival time of the treated mice / mean Survival time of controls χ

d) Evaluated on the basis of findings at autopsy.

EPO COPY

- 13 -

Tabel

IV

Effect against gross leukemia

a)

link Type of
Appointment
reaching " Dose ^b)
(mg / kg) T / C ^c)
% toxic
DeathS ₃ .
cases ^d) DNR iv 10
15th
22.5 142 138
183 185
217 92 2/18
1/18
9/18 example 1 iv 10
15th
22.5 217
150
100 0/10
6/9
9/9 Example 3 iv 6.6
10
15th 185
115
77 - 0/10
7/10
10/10 orally 6.6
10
15th 154
169
92 0/10
0/10
7/8 DX iv 10
13th
16.9 183
225
242 0/10
0/10
0/10 Example 2 iv 4.05
5.27
6.85 233
258
242 0/10
0/10
3/10 orally 4.05
5.27 114
157 0/10
0/10

a) Experiments were inoculated with C-jH mice, iv 2x10 leukemia cells

b) Treatment iv or orally on day 1 after tumor inoculation

c) Mean survival time of the treated mice / mean Survival time of controls χ

d) Evaluated on the basis of findings at autopsy.

EPO copy -: T

The following examples are intended to explain the invention in more detail.

Example 1: -4'-Deoxy-4'-bromdaunorubicin ~ "(A: X = H, R ₁ = OCH ₃ , R ₂ = Br)

2 g of tetra- (η-butyl) ammonium bromide were added to a solution of 4.0 g__4 _ ^ - Epi-4 ^/ -0-trifluoromethanesulfonyl-N-trifluoroacetyl- ■ -daunorubicin (II: R ₁ = OCH ₃ , R ₃ = COCF ₃ ) in 80 ml of anhydrous methylene dichloride added. After 1 h at room temperature, the reaction mixture was washed with water and the organic phase was evaporated in vacuo. The residue was purified on silica gel using a mixture of methylene dichloride and acetone as the eluant; 3.5 g of 4'-deoxy-4'-bromo-N-trifluoroacetyldaunorubicin were obtained, melting point 130 ^° C .; FD - MS 685 (M ⁺ ), TLC on silica gel plates (Merck F 254) using the solvent system methylene dichloride / acetone (10/1 based on the volume) Rf = 0.5.

160 ml of 0.1 μm aqueous sodium hydroxide solution were added to a solution of 3 g of 4'-deoxy-4'-bromo-N-trifluoroacetyldaunorubicin in 20 ml of acetone. After 4 h at 0 ^° C., the solution was adjusted to pH 8.6 with 0.1 η hydrochloric acid and extracted with methylene dichloride. The solvent was evaporated to give a residue which was converted to the hydrochloride by treatment with methanolic hydrogen chloride (2.2 g), m.p. 18O ⁰ C with dec., TLC on Kieselgel plates (Merck F 254) using the solvent system methylene dichloride / methanol / water / acetic acid (80/20/7/3 based on the volume.) Rf = 0.32.

Example 2: 4'-Deoxy-4'-bromodoxorubicin

(A: X = OH, R ₁ = OCH ₃ , R ₂ = Br)

2 g of 4'-deoxy-4'-bromdaunorubicin, prepared as in example 1, were dissolved in a mixture of methanol and dioxane. The solution was, as in U.S. PS

3 803 124, first treated with bromine, the

14-bromo derivative was obtained, and then with aqueous sodium formate, whereby 4'-deoxy-4'-bromodoxorubicin was obtained.

This was converted into the hydrochloride by treatment with methanolic hydrogen chloride, m.p. 170 ⁰ C (decomp.), FD-MS 605 (M ⁺ ), -ILC_auf-Kieselgel-Platten (Merck F 254) using the solvent system methylene dichloride / methanol / water / acetic acid (80/20/7/3 based on the Vol.) Rf = 0.20. ^ -

Example -3: 4'-Deoxy-4'-chlorodaunorubicin

(A: X = H, R ₁ = OCH ₃ , R ₂ = Cl)

Treatment of 4'-epi-4'-0-trifluoromethanesulfonyl-N-trifluoroacetyldaunorubicin (II: R = OCH ₃ , R ₃ = COCF ₃ ) with tetra (n-buty1) ammonium chloride according to the procedure described in Example 1 gave 4'-Deoxy-4'-chlorodaunorubicin, m.p. 175 ° C (dec.), TD-MS 545 (M ⁺ ), TLC on silica gel plates (Merck F 254) using the solvent system methylene dichloride / methanol / water / acetic acid (80 / 20/7/3 based on the volume) Rf = 0.32.

Example 4: 4'-Deoxy-4'-chlorordoxorubicin

(A: X = OH, R ₁ = OCH ₃ , R ₂ = Cl)

Following the procedure described in Example 2. mp 4'-deoxy-4'-deoxy-4'-chlordaunorubicin in 4'-chlordoxorubicin transferred, which was isolated as hydrochloride. 180 ⁰ C (dec.), Mass spectrum FC-561 ( M ⁺ ), TLC on silica gel plates (Merck F 254) using the solvent system methylene chloride / methanol / water / acetic acid (80/20/7/3 V / V) Rf = 0.2.

Example 5: 4'-Deoxy-4 ^/ -fluorodaunorubicin

(A: X = H, R = OCH ₃ , R ₂ = F)

Described in U.S. Patent No. 4,345,068, in 650 ml anhydrous methylene dichloride and 32 ml of anhydrous pyridine, cooled to ⁰ ° C to a stirred solution of 26 g of 4'-epi-N-trichloräthoxycarbonyldaunorubicin, was added a solution of 11 ml of trifluoromethanesulfonic acid anhydride added in 140 ml of anhydrous methylene dichloride. The reaction mixture was with

EPO. COPY

a cooled 5% aqueous sodium bicarbonate solution, ,, - water, -0.1N hydrochloric acid and water in the order given washed. The-over-anhydrous sodium sulfate

- Dried organic solution was used in the next step without further purification used.

A solution of 2 g of 4'-epi-4'-0-trifluoromethanesulfonyl-N-trichloroethoxycarbonyldaunorubicin in 30 ml of acetonitrile was treated with 0.5 g of cesium fluoride. After 10 minutes at room temperature, the mixture was poured into water and extracted with methylene dichloride. The organic phase, dried over anhydrous sodium sulfate, was evaporated to dryness in vacuo. The residue, dissolved in 30 ml of ethanol, was treated with 4 ml of acetic acid and 0.2 g of zinc dust at 0 ^{° C.} After 1 hour, the mixture filtered on Celite was diluted with water and extracted with methylene dichloride.

The crude product obtained by evaporation of the solvent in vacuo was purified by chromatography on a silica gel column using methylene dichloride as the eluent to give pure 4'-deoxy-4'-fluorodaunorubicin, which was isolated as the hydrochloride, FD-MS 529 (M ⁺ ), TLC on silica gel plates (Merck F 254) using the solvent system methylene chloride / methanol / water / acetic acid (80/20/7/3 based on the volume) Rf = 0.32.

Example. 6: 4'-Deoxy-4'-fluorordoxorubicin (A: X = OH, R ₁ = OCH ₃ , R ₂ = P) According to the procedure described in Example 2, 4'-Deoxy-V'-fluorodaunorubicin in 4'- Deoxy-4'-fluorodoxorubicin, which was isolated as the hydrochloride.

EPO COPY

Claims (22)

40 537 u / wa FARMITALIA CARLO ERBA S.P.A. IN MILAN (ITALY) 4'-haloanthracycline glycosides »Process for their production and pharmaceutical preparation containing them Patent claims: 1.) Anthracycline glycoside of the general formula (A) COCK ₂ X EPC CC-PY wherein X is a hydrogen atom or a hydroxy group, R _{1 is} a hydrogen atom or a hydroxy or methoxy group and R _{2 is} a bromine, chlorine or fluorine atom, or a pharmaceutically acceptable acid addition salt thereof 2. 4'-Deoxy-4'-bromodaunorubicin. 3. 4'-Deoxy-4'-chlorodaunorubicin.
4: 4'-Deoxy-4'-fluorodaunorubicin. 5. 4'-Deoxy-4'-bromodoxorubicin. 6. 4 "-deoxy-4'-chlorodoxorubicin. 7. 4'-Deoxy-4'-fluorodoxorubicin. 8. A process for the preparation of an anthracycline glycoside of the general formula (A), wherein R ₁ and R _{2 have} the meaning given in claim 1 and X is a hydrogen atom, characterized in that an exchange reaction between a compound of the general formula (II ) COCH. OH (II), CF ₃ SO ₂ O, wherein R _{1 has} the above meaning and R _{3 is} an amino protective group, and a fluoride, bromide or iodide carries out and of the obtained compound of the general formula (III) B- 3428943 COCH. (Ill) removed the protecting group. 9. The method according to claim 8, characterized in that cesium fluoride is used as the fluoride. that 10. The method according to claim 8, characterized in that the bromide used is tetra (n-butyl) ammonium bromide. 11. The method according to claim 8, characterized in that tetra (η-butyl) ammonium chloride is used as the chloride. 12. The method according to any one of claims 8 to 11, characterized in, that a trichloroethoxycarbonyl group is used as the amine protecting group. 13 .. The method according to claim 12, characterized in that the protective group is removed by hydrogenation. 14. The method according to claim 12 or 13, characterized in that dai the removal of the protective group with zinc dust and acetic acid is effected. 15. The method according to any one of claims 8 to 11, characterized in that that the amine protecting group is a trifluoroacetyl group begins. EPO COPY Ä 16. The method according to claim 15, characterized in that the protecting group is removed by mild alkaline hydrolysis. _. 17. The method according to claim 15 or 16, characterized in that that the removal of the protective group is effected with 0.1N aqueous sodium hydroxide solution. 18. The method according to any one of the preceding claims, characterized in that _daß | _.the exchange reaction in methylene dichloride or acetonitrile is carried out. 19. Process for the preparation of an anthracycline glycoside of the general formula (A), in which R. and R _{2 have} the meaning given in claim 1 and X is a hydroxyl group, characterized in that an anthracycline glycoside of the formula (A) in which R ₁ and R _{2 have} the above meaning and X "represents a hydrogen atom, reacted with bromine and treated the 14-bromo derivative obtained with an aqueous sodium formate solution. 20. The method according to claim 8, essentially as described in one of Examples 1, 3 and 5. 21. The method according to claim 19, essentially as described in one of Examples 2, 4 and 6. 22. A pharmaceutical composition which is an anthracycline glycoside according to any one of claims 1 to 7 or a pharmaceutical acceptable acid addition salt thereof in admixture with a pharmaceutically acceptable diluent or carrier. EPO copy 0m