CS236682B2 - Manufacturing process of 3-deamine analogs of glycoside antheracyclic antibiotics - Google Patents

Abstract

Prodn. of anthracycline glycosides of formula (I) [R1=H or OH; R2=(II) or (III) comprises (a) reacting the corresp. 3',4'diepi- daunorubicin(IV;R1=H) or 3', 4'-diepi-doxorubicin(IV;R1=OH) with NaNO2 and aq. acetic acid to form the mixt. (I) [R1=H, R2=(II) or (III) or the mixt. (I) [R1=OH, R2=(II) or (III) ; and (b) separating the mixt. by chromatography on silica gel after eluting with chloroform and then with 97:3 by vol. chloroform and acetone mix. (I) are useful antitumour agents.

Description

The invention relates to a novel class of glycoside anthracycline antibiotics which are derived from daunorubicin and doxorubicin, to a process for the preparation of these compounds, to pharmaceutical compositions containing them and to the use of these compounds in the treatment of certain tumors in mammals.

Daunorubicin and doxorubicin, i.e. the compounds from which the compounds of the invention are derived, are known and used antitumor antibiotics, which are commonly described in the literature.

In addition, the starting compounds from which the compounds of the present invention are prepared, i.e. 3 ', 4'-diepeptununorubicin and 3 *, 4'-dlepidoxorilicin, are also commonly known compounds, such as are disclosed, for example, in U.S. Pat. No. 4,112,076, Sep. 5, 1978.

The present invention relates to a new class of anthracycline glycosides of formula I:

in which it means

R 1 is hydrogen or hydroxy; and

R 2 is an α-L-pyranoxyl radical of formula (II) or (III):

These anthracycline glycosides contain a neutral glycoside moiety, i.e., either 6-ε-trideoxy-α-L-glycero-hexopyrazin-4-ulosyl (or α-L-cineruloxy) of formula II, or 2,6-dideoxy-α-L-hexypyranosyl (III), marked as follows:

(of formula I wherein R 1 = H, R 2 is equal to II), referred to herein as a compound of formula I-A,

2. 3‘-Deamino-3‘-hydroxy-4‘-epi-daunorubicin (of formula I wherein R 1 = H, R 2 = III), herein referred to as a compound of formula I-B,

3. 3'-Deamino-4'-dehydro-doxorubicin (of formula I wherein R 1 = OH, R 2 equals II), herein referred to as a compound of formula I-C, and

4th 3 ^{<deamino-3'-hydroxy-4'-epi-doxorubicin} (formula I wherein Ri = OH, R2 = III), referred to herein as compound of formula IC.

The present invention provides a process for the preparation of enthracycline glucosides. These novel compounds are prepared from 3 ‘, 4‘-diepidaunorubicin (Formula IV wherein R 1 is H) and 3‘, 4‘-dipdoxorubicin (Formula IV wherein R 1 = OH). Both of these compounds are commonly known anthracycline glycosides, which are exemplified in the aforementioned U.S. Patent No. 4,112,076 of September 5, 1978, and are represented by the following formula:

The process for the preparation of the novel compounds of the invention comprises the deamination of anthracycline glycosides of formula IV in which the axial amino group is adjacent to an equatoreal hydroxy group running through the stained intermediate to give formulas IA and IC as a result of hydrogen migration from C-4 '. and ID by a simple nucleophilic reaction of water at the C-3 'position.

The process according to the invention is carried out by reacting an aqueous solution of 3 ', 4'-diethyldororubicin hydrochloride or 3', 4'-diepidoxorubicin hydrochloride of the above general formula (IV), wherein R 1 is hydrogen or hydroxy, separately reacted at 0 ° C. from 3 to 4 hours with sodium nitrite and 1N aqueous acetic acid to give a 3'-aminated glycoside mixture of formula (I), i.e. a mixture of 3'-deamino-4'-dihydrot daunorubicin of formula (I); wherein R 1 is hydrogen and R 2 is a group of formula II and 3'-deamino-3'-hydroxy-4'-epi-daunorubicin of formula I wherein R 1 is hydrogen and R 2 is a group of formula III, or a glycoside of formula I, i.e. a mixture of 3'-deamino-4'-dehydro-doxorubicin of formula I, wherein R 1 is hydroxy and -R 2 is a group of formula II and 3'-deamino-3'-hydroxy-4'- of formula (I), wherein R 1 is n and hydroxy represents a group of formula (III), each mixture being subjected to chromatography on a silica gel column, eluting first with chloroform and then with a 97: 3 mixture of chloroform and acetone, each of which compounds of formula I are obtained as the free base in pure form.

Said deamination reaction is preferred. The embodiment of the invention is carried out with sodium nitrite in a cold aqueous acidic medium, but can also be carried out with nitrous oxide or alkyl nitrite in a cold aqueous medium or in an alcoholic medium.

The novel anthranil glycosides produced according to the invention exhibit an antitumor effect. In particular, 3‘-deamino-3‘-thydroxy-4‘-epitdoxorubicin of the formula I-D, which exhibits a significant effect on artificially induced tumors in mice, may be mentioned. For this reason, the invention also relates to pharmaceutical compositions comprising a therapeutically effective amount of an anthracycline glycoside of formula I which is admixed with a pharmaceutically acceptable diluent or carrier, and to a method of treating certain tumors in a mammal comprising administering to the affected patient a therapeutically effective amount. of the compounds of the invention.

The invention will be further illustrated by the following examples with reference to the results of biological tests.

Example 1

3‘-Deamino-4‘-dihydro-daunoгubicin of formula I-A and 3‘-deamino-3‘-hydroxy-4‘-epidaunirubin of formula I-B

A solution containing 0.55 g (1 mmol) of 3 ', 4'-di epi-daunorubicin hydrochloride - (general formula IV, R = H) in 25 mL water was cooled to 0 ^Q C and then treated with 0 72 (7.5 mmol) of sodium nitrite and 7.5 ml of a 1 N aqueous acetic acid solution are added in several portions over 20 minutes with stirring, the rate of addition being such that the reaction temperature does not exceed 0 ° C. After 3 hours while maintaining the mixture at 0 ° C, the reaction mixture containing the red precipitate was allowed to warm to room temperature. Nitrogen is then bubbled through the solution to remove excess nitrous acid, and then the solution is extracted with chloroform. The chloroform extract was washed with water, dried over anhydrous sodium sulfate, evaporated to a small volume and subjected to chromatography on a silica gel column. Elution with chloroform gave 3'-deamino-4'-dehydro-daunorubicin (IA) as an amorphous solid, m.p. 143-144 ° C (dec.).

[Α] D ²³⁰ = + 190 ° (c = 0.05, in methanol).

UV and VIS spectrum - λ CHsOH:

235, 253, 290, 480, 496 and 530 nm, (E] 808, 557, 188, 236, 242 and 143).

Desorption Mass Spectrum: m / z 510 (M &lt; + &gt;), 398 and 562.

1 H-NMR spectrum (CDCl 3):

1.36 (d, J = 6.5 Hz, 3H, C H 3 O 3-5), 1.70-2.40 (m, 6H, H-8, H-2 ‘, H-3 3),

2.41 (s, 3H, COCH 3),

2.97 (d, J = 19.5 Hz, 1H, H-10 _ax)

3.25 (dd, J = 19.5 Hz, 1 H, H-10 _ak ),

4.09 (s, 3H, OCH 3),

4.40 (q, J = 6.5 Hz, 1H, H-5 5),

4.62 (s, 1H, OH-9);

5.41 (m, 1H, H-7);

5.67 tt, J = 6.0 Hz, 1H, [delta],

7.30-8.10 (m, 3H, aromatic protons), 13.29, 14.01 δ (two s, 2H, OH-6, OH-11).

Further elution with 97: 3 chloroform: acetone gave 3'-deamino-3'-hydroxy-4'-epi-daunorubicin (IB), m.p. 165-170 ° C. (with decomposition).

[.alpha.] _{D @} 20 = + 350 DEG (c = 0.025, in methanol).

UV and VIS spectrum - λ ™ ° н:

235, 254, 290, 480, 496, and 530 nm, _{( .lambda.} = 714, 509, 154, 226, 231, and 142).

Desorption Mass Spectrum: m / z 528 (M &lt; + &gt;), 398, 362.

1 H-NMR spectrum (CDCl 3):

1.36 (d, J 1 = 6.0 Hz, 3H, CH 3 -5 ‘), δ σ σ ^ (m, 4H, H-8, H-2‘),

2.41 (s, 3H, COCH 3),

2.86 and 3.30 (two d, J = 19 Hz, 2H, H-10),

3.5-4.0 · (m, 2H, H-3 ', H-4'),

4.07 (s, 3H, OCH 3),

4.65 (m, 1H, H-5 '),

5.25 (m, 1H, H-7);

5.48 (m, 1H, H-1 H),

7.30-8.10 (m, 3H, aromatic protons),

13.26, 13.98 δ (two s, 2H, OH-6, OH-11).

She said d ^

3‘-Deamino-3‘-dehydro-doxorubicin of formula I-C and ddeamino-dhydroxy-4-epi-doxorubicin of formula I-D

The above compounds of formulas I-C and I-D were prepared from 3 ‘, 4‘-die, β-doxorubicin (IV, R 1 = OH) in the same manner as in Example 1.

There was obtained 3‘-deamino-3‘-dehydro-doxorubicin of formula I-C as a red powder, m.p. 140 ° C (dec.).

[.Alpha.] D @ 20 = +145 (c = 0.046, methanol).

UV and VIS spectrum - A ° ”OH:

235, 254, 290, 480, 496 and 530 nm, _{( εm} 690, 490, 190, 228, 230, 140 ₎ .

Desorption mass spectrum:

m / z 526 (M &lt; + &gt;), 414, 336.

1 H-NMR spectrum (CDCl 3):

14.9 (C-6),

27.6 (C-2),

33.1 (C-3),

33.9 (C-10 ·)

35.9 (C-8),

56.6 (O — CH3)

65.4 (C-14)

68.9 (C-7),

71.3 (C-5),

76.6 C-9

100.3 (C-1),

118.5 C-3

119.8 (C-1, C-aa)

133.7 (C-6a, C-10a, C-12a)

135.8 C-2

155.6 (C-11),

156.1 (С-б ,.

161.0 (C-4),

186.9 (C-5, C-12)

210.3 (C-4),

213.98 (C-13).

In a similar manner, the 3‘-deamino-4-hydroxy-4-epi-doxorubicin of formula I-D is obtained as a red powder, m.p. 176 DEG-180 DEG C. (with decomposition).

(a) _{D 20} = + 280 ° (c = 0) 046) in methanol).

UV and VIS spectrum - A *:

235, 254, 290, 480, 496 and 532 nm, (E 1% = 674) 478, 186, 223, 228 and 148).

Desorption mass spectrum:

m / z 544 (M &lt; + &gt;), 414, 378, 336.

и ш t

¹³ C-NMR (CD 3 OD: CDCl 3, 1: 1 v / v):

17.8 (C-6), -33.9 (C-10),

36.3 (C-8),

3.80 (C-2),

57.0 (СНз),

65.4 (C-14),

68.8 (C-7),

69.8 (C-3 ‘, C-4‘)

77.0 (C-9)

77.9 (C-5),

101.4 (C-1), Daunorubicin. HCl 12 119.3 (C-3, C-4a), Compound I-A 350 120.3 (C-1), Compound I-B 60 134.3 and 135.8 (C-6a, C-10a, C-12a), Doxorubicin. HCl 11 136.5 (C-2), Compound I-C 40 155.7 (C-6, C-11), Compound I-D 25

Biological efficacy

The cytotoxic activity of the novel anthracycline glycosides of the present invention was tested in a conventional manner against

Не La cells in vitro (duration of action of compounds 24 hours) compared to daunorubicin and doxorubicin. The results obtained are shown in Table 1.

TABLE 1

In Vitro Colony Growth Inhibition Assay

Compound

ID 50 (ng / ml)

Certain compounds of the invention exhibit excellent results when tested against P-388 and L1210 leukemias in mice compared to those obtainable with daunorubicin and doxorubicin, especially when administered orally. The results regarding the antitumor effect of the compounds of the invention against various types of leukemias in mice are shown in Tables 2, 3 and 4 below.

TABLE 2

Anti-tumor effect in leukemia P 388. Intraperitoneal treatment on the first day

Compound Dose (mg / kg) T / C ^and (%) LTS ^b Toxic Death® Daunorubicin ^d 2.9 170, 150 0/9, 0/10 0/9, 0/10 4.4 154,150,155 0/8, 0/9, 0/10 2/8, 3/9, 0/10 6.6 145, 150,160 0/8, 0/9, 0/10 7/8, 5/9, 0/10 Compound IA ^d 30.8 109 0/8 0/8 30 127 0/8 0/8 60 160, 165 0.3, 0/10 0/3, 2/10 78 95 0/10 5/10 101 100 ALIGN! 1/10 5/10 Doxorubicin® 4.4 170 0/10 0/10 8.6 160 0/10 6/10 Compound IB ^e 15 Dec 180 0/5 0/5 20 May 290 0/5 0/5 26 160 0/5 3/5 Doxorubicin ^e 4.4 225 0/8 0/8 6.6 290 · 1/8 0/8 10 300 2/8 0/8 Compound IC ^e 10 170 2/10 0/10 15 Dec 180 0/8 0/8 22.5 210 0/8 0/8 33.7 230 1/8 0/8 Doxorubicin * 4.4 245, 223 0/10, 0/10 0/10, 0/10 6.6 255, 258 1/10, 0/10 0/10, 0/10 10 245, 282 2/10, 1/10 0/10, 0/10 Compound ID ^f 6.6 250 1/10 0/10 10 290 3/10 0/10 15 Dec > 630,> 658 6/10, 6/10 0/10, 1/10 22.5 > 630,> 658 9/10, 5/10 0/10, 2/10 33.7 194 3/10 5/10 50.5 76 1/10 7/10

Notes to Table 2:

^and mean survival time, in ° / o compared to control experiments ^b long-term survival (at least 60 days) ^c as toxic death in Table 2 to 4 denotes death due to the toxic action of the substance; based on autopsy findings in dead mice ^d values from three samples ^e substance dissolved in 10% surfactant solution (Tween 80) ¹ substance dissolved or suspended in water, values from two experiments.

TABLE 3

Effect of Compound I-D on Leukemia in Mice

Tumor Way Way Day Treatment Dose (mg / kg) T / C ^and (%) LTS ^b Toxic. death Environment Compound Grossův iv iv 1 H2O Doxorubicin 10 200 0/8 0/8 13 241, 275 0/18 1/18 16.9 233, 175 1/18 6/18 Compound 13.3 183 0/7 0/7 I-D 26.6 225 0/8 0/8 39.9 250 0/8 3/8 iv oral 1 Tween ^e Compound 26 133 0/7 0/7 I-D 33.9 158 0/10 0/10 44 158 0/10 0/10 L 1210 ip iP 1 Tween ^e Doxorubicin 4.4 166 0/10 0/10 6.6 166 0/10 0/10 10 177 3/10 0/10 Compound 8.8 > 600 3/6 0/6 I-D 13.3 505 3/8 0/6 20 May 238 0/8 2/8 iv iv 1 Tween ^e Doxorubicin 10 157 0/10 0/10 13 178 0/10 0/10 16.9 185 0/10 0/10 Compound 13.3 185 0/9 0/9 I-D 20 May 264 0/9 0/8 30 600 3/8 0/8

Note:

^a - ^b - ^c have the same meaning as in Table 2 ^d of the two experiments ^e Tween 80 surfactant, 10% solution in water.

TABLE 4 Effect of Compound I-C on Gross Leukemia Compound Environment Method Dose T / C ^and (%) LTS ^b Toxic (mg / kg) death ^c

Doxorubicin Mon 80 ^d iv 13 ^e 16.9 ^e 214,275,242 228, 175, 250 0/20 1/28 0/20 8/28 Compound I-C Mon 80 ^d iv 40 ^f 185, 250 0/17 2/17 48 116 0/10 7/10 57.6 116 0/10 10/10 Compound I-C Mon 80 ^d orally 40 192 0/8 0/8 48 200 0/8 0/8 57.6 200 0/8 2/8

Notes to Table 4:

^a · ^b - ^c have the same meaning as in Table 2 ^d have the same meaning as in Table 3 ^e values from three experiments ^f values from two experiments.

It is within the scope of the invention that certain modifications and alternatives may be made without departing from the scope of the invention.

Claims (1)

A process for the preparation of 3 dea-deaminated analogues of glycoside anthracycline antibiotics of general formula I wherein: R1 is hydrogen or hydroxy; R2 is a group of formula II (II) or III (also characterized in that an aqueous solution of 3 ', 4'-diepiedonororubicin hydrochloride or 3', 4'-dieeddoxorubicin hydrochloride of formula IV í ^ íl OH ιίπ COCH ₂ fy-OH | H ^ H ^CO 0 он 0 / riC— Λ J 7 («in) но-T- where Rt is hydrogen or hydroxy, separately reacted at 0 ° C for 3 to 4 hours with sodium nitrite and 1 N aqueous acetic acid to give a 3'-deaminated mixture of glycoside of formula I, i.e. a mixture of 3'-deamino-4'-dehydrodaunorubicin of formula I wherein R1 is hydrogen and R2 is a group of formula II and 3'-amino-3'-hydroxy-4'-epi-daunorubicin of formula I wherein R1 is hydrogen and R1 is a group of formula (III) or a glycoside of formula (I), i.e. a mixture of 3'-deamino-4'-dehydro-doxorubicin of formula (I) wherein R1 is hydroxy and R2 is a group of formula (II); 3'-deamino-3'-hydroxy-4'-epi-doxorubicin of formula I, wherein R 1 is hydroxy and R 2 is III, each mixture is chromatographed on a silica gel column, eluting first with chloroform and then with chloroform And acetone in a volume ratio of 97: 3, each of said compounds of formula (I) being obtained as the free base in pure form.