CS273199B2 - Method of epidophyllotoxine-glucoside's derivatives production that contain nitrogen - Google Patents

Abstract

A method of production of compounds of general formula IV, where B represents the residue of formula IVa, IVb or IVc, where R<11> is the group -C(O)H, -C(O)R<10>, -C(O)OR<10> or -C(O)NHR<10>, where R<10> represents C1-5-alkyl possibly substituted with 1 to 3 halogens nebo phenyl, significant in the fact that the compound of general formula VI is oxidised causing the occurrence of a compound that is possibly subjected to reduction and cleavage of the acyl group. The compounds inhibit the growth of tumours and can be used in medicine.<IMAGE>

Description

The present invention relates to novel nitrogen-containing epipodophyllotoxin-glucoside derivatives, to a process for their preparation, to their therapeutic uses for combating cancer and to pharmaceutical compositions containing these novel substances.

Etoposide (VP-16, Formula Ia) and teniposide (VM-26, Formula Ib) are clinically used anti-tumor agents derived from the natural lignan of podophyllotoxin of formula II

The numbering of positions for nomenclature purposes is also indicated in formula II,

Etoposide and Teniposide They are derivatives of epipodophyllotoxin, where epipodophyllotoxin is the epimer of podophyllotoxin in item 4. Etoposide They are effective in the treatment of a number of cancers including small cell lung, non-lymphocytic leukemia and varletaneseminoma type tumors (ama Drug Evaluation, 5th ed. 1983, Chicago, Illionis, pp. 1554-1555).

CS 273 199 B2

Etoposide and tenipoeid as well as methods for their preparation have been described by Keller-Ouslen et al. U.S. Patent No. APLA. 3524 844. Etopoeid-3I ^# 4 -chinon disconnects / idajíci formula below IIIa formed in the electrochemical oxidation of etoposide £>. □. M. Holthuis et al.,

3. Electroanal. Chem, Interfacial Elektrochem., 1985, 184 (2), 317-3291. Ooeef German in U.S. Patent No. 4,609,644 describes the preparation of a quinone of formula III

IIIa »CH ₃ ; Y »H

Ayres and Lim in Cancar Chamother, Pharmacol., 7, 88-101 (1982) describe podophyllotoxin corresponding to the formula

(1)

The invention provides antitumor active compounds of formula IV

CS 273 199 B2

in which

B is a radical selected from the group consisting of radicals of formulas. IVa, IVb and IVc

(IVa) where, _R 11

represents a radical -CH, -CR ¹⁰ , n ii oo wherein R ¹⁰ is an alkyl group having 1 to 5 1 to 3 halogen atoms or an phenyl group

-COR ¹⁰ or -CNHR ¹⁰ ,

II by o carbon atoms, group.

(IVb) optionally substituted

By alkyl groups is meant straight or branched saturated carbon chains, such as methyl, ethyl, π-propyl and isopropyl. Halogen atoms are fluorine, chlorine, bromine or iodine atoms.

The invention also provides pharmaceutical compositions comprising an antitumor compound of formula IV and a pharmaceutically acceptable, non-toxic carrier.

CS 273 199 B2

The invention further provides a method of inhibiting the development of tumors in a mammal comprising administering to a mammal with such a tumor an effective amount of an antitumor active compound of formula IV.

It is an object of the present invention to provide a process for the preparation of compounds of the above formula (IV), characterized in that the compound of the formula (VI)

oxidizes to form a product of formula IV, wherein B is a radical of formula IVe, which is optionally subjected to reduction and cleavage of an R ¹¹ radical.

The process according to the invention thus consists in modifying a residue in the meaning of the symbol B and can be described by the following scheme I i

OH

OH .5 .CS 273 199 B2

Oxidation of the compound of formula (VI) may be accomplished using a combination of sodium periodate, acetonitrile and water, or a combination of sodium nitrite, acetic acid and tetrahydrofuran, or other conventional oxidizing agents.

The reduction of the quinone residue of formula XVa to the hydroquinone residue of formula IVb can be carried out by catalytic hydrogenation or by treatment with a suitable reducing agent such as sodium metabisulphite.

The acyl group R @ 1 may be cleaved by conventional methods.

For example, the trichloroethoxycarbonyl group can be cleaved by zinc and acetic acid.

The starting materials of the general formula (VI) can be prepared as described in our related Czechoslovak patent specification No. P 273 185.

In particular, the aforementioned antitumor activity is shown by the compounds of formula (IV) above, in which B is a radical of formula (IVa). This efficiency is demonstrated by the following tests.

Biological activity

The antitumor activity of representative compounds of the invention was evaluated in vitro for cytotoxicity against human and mouse tumor cell lines. As well as transplantable mouse leukemia P388.

Leukemia P3 88

Female (CDFα) mice are intraperitoneally implanted with 10 aecitic tumor cells of P388 leukemia mouse and animals are dosed with different doses of the test compound. Each dose is tested in 4 mice and 10 animals serve as control (given saline only). Test substances aa injected intraparitoneally on days 5 and 8 (day 1 is considered the day of tumor implantation). Antitumor activity is expressed as T / C (%), which is a percentage of the mean survival of the drug treated group to the mean survival of the control group. A compound having a T / C (%) ratio equal to or greater than 125 is generally considered to have significant anti-tumor activity in the P388 leukemia assay. The experiment lasts for 31 days and after the end of the experiment it records the number of surviving animals.

The results of the test described above are shown in Table I below. only the maximum T / C (%) and the dose at which the maximal effect occurs.

Table I

Anti-tumor efficacy against murine P388 leukemia compound of the dose of Example no.

(mg / kg) ^x ) maximum value

T / C (%) ^x

160 (25) 2,200 (80)

124 (> 140)

135 (275)

CS-273 199 B2

Legend:

x) The values in brackets represent the gain data using etoposide as a positive control in the same experiment

Cytotoxicity test

Teet cytotoxicity in vitro consists in culturing a variety of mammalian tumor cells, including human tumor cells, in microtiter plates using conventional tissue culture methods. Using the quadruplicate dilution technique, the concentrations required to inhibit cell growth by SO% (ICg ₀ ) are determined for each compound. The validity of this method is confirmed by a report published in the Proceading of the American Association for Cancer Research, 1984, 25, 1891 (Abst. 328). Tumor cells of the following types are used for each test compound:

B16-F10,

Moeer colon cancer, SW900 human lung cancer and three human colon cancer cells, HCT-116, HCT-VN and HCT-VP, the latter two being resistant to teniposide (VM and ethoposide VP, respectively).

IC _5Q less than 500 .mu.g / ml are a positive indicator of antitumor activity.

IC ₅₀ values of various compounds of the invention against the above cell lines are shown in Table II below.

Table XX

IC ₅₀ values (µg / ml) found in the in vitro cytotoxicity assay of compound z

Example no. B16-F10 HCT-116 HCT / VM34 HCT / VP35 MOSER SW900 (1) 72 66 58 84 87 80 1 81 68 64 70 71 > 188 (X) 39 53 55 77 71 ) 188 '2 38 46 53 79 77 > 188

Legend:

(x) values for etoposide in the same experiment are 2.7, 1.9 (B1S-F10), 2.1, 2.7 (HCT-116), 6.1, 3.1 (HCT / VM34), 30 41 (HCT / VP35). 39, 39 (Moser) and 67, 12.5 (SW900) (1) values for etopoaid in the same experiment are 7.0, 4.6 (B16-F10 (, 9.6, 10.2 (HCT116)), 31, 33 (HCT / VM34), 92,51 (HCT / VP35), 126,112 (Moser) 8-25,65 (SW900)

The above animal tests show that the compounds of formula (IV) act as potent inhibitors of tumor growth in mammals. Accordingly, the invention also provides a method for inhibiting tumor growth in mammals, comprising: and administering to said mammal suffering from a tumor an effective inhibitory dose of an antitumor active compound of formula (IV).

The invention further provides pharmaceutical compositions comprising an effective inhibitory amount of an antitumor active compound of formula (IV) and a pharmaceutically acceptable carrier. These compositions may be formulated into any dosage form suitable for the particular mode of administration. Examples of such compositions include solid compositions for oral administration, such as tablets, capsules, pills, powders and granules, liquid compositions for oral administration, such as solutions, suspensions, syrups and elixirs, and formulations for parenteral administration. As sterile solutions, suspensions and emulsions. The present compositions can also be prepared as sterile solid preparations which can be dissolved in sterile water, physiological saline solution, or other sterile injectable media prior to use.

Choosing appropriate dosages and dosing regimens for the mammal to be treated will not cause problems to those of skill in the art. It will be appreciated that the dosage employed will vary depending upon the nature of the composition employed, the active ingredient, the mode of administration, the patient being treated, the patient being treated and the type and severity of the disease. agents such as age, weight, sex, nutrition, time of administration, route of administration, rate of excretion, patient condition, combination of active ingredients, patient sensitivity to the active agent, and severity of the disease.

The invention is illustrated by the following examples, which are not intended to limit the scope of the invention in any way.

The melting points reported were determined on a capillary melting point (Thomas-Hoover) and are not corrected. ¹ H-NMR spectra were measured on either a Bruker WM 360 or Varlan VX2 200 spectrophotometer using deuterochloroform as an internal standard). Chemical shifts are given in values of J of the coupling constant in Hz. Signal shapes are denoted by the usual abbreviations 8 (einglet), d (doublet), t (triplet), q (quota), m (multiplet), b (broad signal) and dd (doublet of doublets). IR spectra were measured on either a Beckman Model 4240 or a Perkin-Elmer 1800 Fourier Transform Infrared Spectrophotometer, and are reported in cm- ^X . Thin layer chromatography was performed on finished silica gel plates (60F-254) with detection by UV radiation and / or iodine vapor. High and low resolution mass spectroscopy was carried out on KRATOS MS 50 and MS 50 resp. Kratos MS 25RFA. By flash chromatography, the mini method described by WC Still et al. in 3. Org. Chem., 43, 2923 (1978). This chromatography was ^- BU3 not performed on Merck silica gel (0.074 to 0, 038 mm) or Woelm silica gel (32-63 / ^u m). All solvent evaporations were carried out under reduced pressure.

Example 1

5'-5'-Oemethoxy acetylaminoetopoeid-3 '4'-quinone (IV B »IVa, R ^X1>» acotyl)

A. 3 ^Z- Acetamldoetoposide

To a solution of 0.420 g (0.70 mmol) of 3 ' -aminoetoposide in 14 ml of long dichloromethane and under stirring at d &lt; 2 & gt ^; 0.065 ml (0.69 mmol) of acetic anhydride was added dropwise. The reaction mixture was stirred at 2 ° C for 5.5 hours and then an additional 0.01 mL (0.11 mmol) of acetic anhydride was added. The resulting ββ mixture was stirred at 2 ° C for 1 hour,

CS 273 199 B2

It is then poured into a solution of 25 ml of water and 25 ml of a saturated aqueous sodium bicarbonate solution, extracted three times with 50 ml of dichloromethane each time, the combined organic layers are washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate and concentrated. The residue was flash chromatographed on silica gel using 3% β / 4% methanol in dichloromethane first as eluent to give 19.3 mg of an unidentified by-product (thin layer chromatography in 5% methanol in dichloromethane my Rf 0.17) and 0.328 g (73%, title compound) as an off-white powder, mp 225-227 ° C.

Thin layer chromatography in 5% methanol in dichloromethane gave the product

Rf 0.14 ·

IR (KBr): 3350-3080, 2960, 1770 cm ^1st

¹ H-NMR (CDCl 3) δ 8.45 (β, 1H), 8.23 (bs, 1H), 6.76 (β, 1H),

6.61 (s, 1, H), 6.49 (β, 1H), 6.38 (a, 1H), 5.84 (d, 3 - 8.6 Hz, 2H),

4.86 (d, O 3.2 Hz, 1H), 4.63 (m, 1H), 4.30-4.15 (m, 3H), 4.15-4.05 (m, 2H), 3.72 (a, 3H), 3.6-3.4 (m, 2H), 3.4-3.25 (m, 4H), 2.86 (m,

1H), 2.19 (β, 3H), 1.26 (d, O = 5.0 Hz, 3H).

B. ^5/5 * ^{Desmethoxy--acetamldoetoposid-3/4 Z} - quinone

To a solution of 0.150 g (0.244 mmol) of the above-prepared 3 ^of -ecetemidoetopanol in 10 ml of dry tetrahydrofuran was added under stirring at nitrogen at 2 ° C first.

1.5 ml (26.2 mmol) of acetic acid and then 34 mg (0.49 mmol) of sodium nitrite. The reaction mixture is stirred at 2 ° C for 3.5 hours, then poured into 100 ml of a mixture of equal parts of ethyl acetate and diethyl ether and washed twice with 50 ml of saturated aqueous solution each time. aqueous sodium bicarbonate solution and once with 50 ml of saturated aqueous sodium chloride solution. The organic layer was dried over magnesium sulfate, concentrated in vacuo and the residue subjected to flash chromatography on silica gel using 5% methanol in dichloromethane as eluent. 91 mg (62%) of a dark red solid are obtained. Melting point: 245-250 ° C.

IR (KBr): 3450 (br), 2925, 1779, 1669 cm ^{-1, 1} H-NMR (CDCl <P), 7.78 (m, 2H), 6.77 (s, 1H) 6.56 (s, 1H).

5.99 (m, 2H), 5.70 (e, 1H), 4.89 (d, D 3.24 Hz, 1H), 4.74 (q, 3 5.1 Hz, 1H), 4, 56 (d, δ 7.8 Hz, 1H), 4.45 (t, 0. 3.2 Hz, 1H), 4.56 (d,

7.8 Hz, 1H), 4.45 (t, 3.2 Hz, 1H), 4.34 (t, 7.5 Hz, 1H),

4.26 (d, 3-6.3 Hz, 1H), 4.16 (dd, 3.4, 2.1 Hz; 1H), 3.70 (t, 3 »

8.5 Hz, 1H), 3.57 (m, 1H), 3.42 (T, 3 11.6 Hz, 2H), 3.34-3.29 (m,

2H), 2.89 (m, 1H), 2.75 (bs, 1H), 250 (bs, 1H), 1.37 (d, 3 4.9 Hz, 3H).

Example 2

CC (2-chloroethyl) amino carbonyl] aminoetoposld-3 ', 4'-orthoquinone (IV, B-IVa, R ¹¹ ' - 2- (chloroethyl) amino-3-carbonyl)

A. 3- [(2-Chlorethylamino) kerbonyl] amino-3 ^, -demethoxyathanol

To a solution of 0.40 g (0.69 mmol) of 3'-aminoetoposide in 4 ml of dry dichloromethane was added dropwise under nitrogen at 20 ° C via a syringe 62 µl (0.76 mmol) of 2-chloroethyl-8-cyanate. . Immediately after addition, a white precipitate formed. The reaction mixture is stirred at 20 ° C for 4 hours, then at 0 ° C for 15 minutes and then filtered off with suction. After drying under vacuum, ββ yields 0.32 g (68%) of an off-white amorphous

CS 273 199 B2 solids having a melting point of 195 to 197 ° C.

IR (KBr) 3400 (broad band), 2920, 1770, 1660 cm &lt; 1 &gt; H-NMR (doutorochloroform / dlmethylsulfoxide, cf values):

8.97 (bs, 1H), 7.87 (b, 1H), 6.71 (a, 1H), 6.54 (t, 0-5.6 Hz, 1H),

6.33 (.delta., 1H), 6.31 (.beta., 1H), 6.24 (.delta., 1H), 5.78 (.alpha., 3.5-4.5 Hz, 2H),

4.79 (d, 3 - 5.3 Hz, 1H), 4.56 (m, 1H), 4.51 - 4.24 (m, 4H), 4.06 - 3.98 (m, 2H) 1.61 (a, 3H), 3.45-3.33 (m, 6H), 3.24-3.12 (m, 4H), 2.80 -2.76 (m, 1H), 1.21 (d, 3-2.7 Hz, 1H).

Mass Spec ^. MH + calcd for C 6 HH ^ gOO^NgCl 678, 1828 found 678, 1826.

B. ^{5/5 Desmethoxy-z} - (X (2-chloroethyl) Amino] carbonyl] amino etoposide 3 ^{a, 4,} -orthochi 'non

The general procedure described in Example IA was repeated except that the product prepared in the previous paragraph was used instead of 3'-acetamidoatoposide. The obtained compound melted from decomposition at 195-197 ° C.

IR (KBr) 3400 (broad band), 2940, 1780 cm ^{@ -1} .

1 H-NMR (CDCl 3): 8.62 (β, 1H), 7.53 (s, 1H), 7/43 (s, 1H), 7.28 (bt, 3-7.2 Hz) 1 H, 6.63 (s, 1 H), 6.61 (s, 1 H), 5.98 (bs, 2 H),

5.59 (s, 1H), 4.95 (d, 3-3.1 Hz, 1H), 4.79 (m, 2H), 4.58-4.30 (m,

3H), 4.28-4.13 (m, 2H), 3.80-3.60 (bm, 7H), 3.60-3.25 (bm, 3H),

2.98 (m, 1 H), 1.41 (d, 3 5.1 Hz, 3H),

FAB-MS: m / e 665 (MH ^&lt; ^{+ &gt;} ).

Example 3

^{3 Desmethoxy-5--demothyl 3} -acetylaminoetopo8id (IV-B IBB R ^1J - - acetyl) ·

The procedure described in Example IA was repeated using 65.4 µl (0.69 mmol of acetic anhydride and 0.38 g (0.66 mmol) of 3'-aminoetoposide in 10 mL of dichloromethane.) The product obtained after drying the organic phase with magnesium sulfate and concentrating in Dissolve the vacuum in 10 ml of anhydrous tetrahydrofuran and first add 1.5 ml (26.2 mmol) of acetic acid and then 0.182 g (2.64 mmol) of nitrite nitrite, followed by the procedure described in Example 1B. The solid obtained after flash chromatography was dissolved in 100 mL of ethyl acetate, 60 mL of saturated aqueous sodium metabisulphite solution was added, and the mixture was shaken until the dark red color disappeared (less than 1 minute). The residue was subjected to flash chromatography on silica gel using 4% methanol in dichloromethane as eluent to give 73 mg (18%) of the title compound as a white solid, m.p. %) of the product as a yellow-red solid melting at 210-220 ° C.

CS 273 199 B2

IR (KBr) 3440 (broad band), 2945, 1775 cm @ -1. @ ¹ H-NMR (CDCl3) .delta .: 9.49 (ba, 2H), 6.97 (bs, 1H), 6.84 (β, 1 H),

6.61 (d, O 2.0 Hz, 1H), 6.46 (s, 1H), 6.20 (d, 0-2.0 Hz, 1H), 5.95 (s, 2H), 4 94 (d, 0-3.6 Hz, 1H), 4.75 (q, 0-5.2 Hz, 1H), 4.55-4.39 (m, 3H), 4.30-4, 16 (m, 2H), 3.61 (m, 2H), 3.43-3.29 (m, 4H), 2.98 (bm, 1H), 2.19 (β, 3H), 1.38 (d, 0-4.8 Hz, 3H).

FAB-MS: m / e 602 (MH ^&lt; ^{+ &gt;} ),

Example 4

3'-Demethoxy-5'-demethyl-3'- [[C. (2,2,2-trichloro) ethyl] oxy 3 carbonyl; Amino etoposide 3 (IV, B 'IVb, R ¹¹ £ (2,2,2-trichloroethyl) oxycarbonyl athyl3)

A, 3 ^from -Omethoxy-3-ETC (2,2,2-trichloroethyl) oxy] carbonyl-13-aminoetoposide

To a solution of 0.40 g (0.70 mmol) of 3'-aminoetoposide and 90 µl (1.11 mmol) of pyridine in 5 ml of dichloromethane under stirring at 2 ° C under nitrogen is added dropwise 0.10 ml ( 0.73 mmol) of trichloroethyl chloroformate. The reaction mixture was stirred at 2-10 ° C for 5 hours, then an additional 10 µl of trichloroethyl chloroformate was added thereto, and the reaction mixture was stirred at room temperature for 15 hours.

The resulting mixture was poured into water (50 ml) and extracted with dichloromethane (3.times.50 ml). The combined organic layers were dried over anhydrous magnesium sulfate and purified by flash chromatography on silica gel using 3o (methanol in dichloromethane as eluent). isolation to give 0.32 g (62%) of a yellow solid melting above 220 ° C.

δδ (KBr-technique): 3440, 2910, 1780 (broad band), 1625, 1552 cm ^-1 , ¹ H-NMR (CDCl 3): 7.12 (m, 1H), 6.83 (m, 2H) ), 6.49 (β, 1H), 5.95 (m, 2H), 5.65 (s, 1H), 4.94 (d, 0 »3.28 Ηζ, ΙΗ), 4.80-4 60 (m, 4 H),

4.54 (d, 3 - 5.3 Hz, 1H), 4.38 (m, 1H), 4.23-4.05 (m, 2H), 3.94 (s, 3H),

3.72 (m, 1H), 3.52 (m, 1H), 3.43 (m, 1H), 3.32 (m, 2H), 3.23 (dd, 3-14.2,

5.4 Hz, 1H), 3.01 (m, 1H), 2.68 (s, 1H), 2.41 (s, 1H), 1.38 (d, 3-2.4 Hz, 3H) .

FAB-MS: m / e 748 (M + H) &lt; + & gt ^; .

B.3 ^/ -0emethoxy 5 ^of -demothyl-3 ^/ - CCC (2,2,2-trichloro) ethyl] oxy J -karbonyl3 amino etoposide 3

To a solution of 0.354 g (0.473 mmol) of the above prepared etoposide-5 ^from -trichloroethylcarbamate and 3 ml of acetic acid in 10 ml of tetrahydrofuran was added under stirring at nitrogen at 2 ° C 0.200 g (2.90 mmol) of sodium nitrite. The reaction mixture was stirred for 3 hours, then the resulting red solution was poured into 80 mL of saturated aqueous sodium bicarbonate and extracted first with 80 mL of ethyl acetate and then twice with 25 mL of ethyl acetate. The combined organic layers are washed with 50 ml of aqueous sodium chloride solution, dried over sodium sulphate and concentrated in vacuo to a red solid which is dissolved in 7 ml of tetrahydrofuran. To the solution is added 1 ml of water and 1 ml of glacial acetic acid. The mixture was cooled in ice water and 0.20 g (3.06 mmol) of activated zinc was added. The reaction mixture is then removed from the cooling bath and the red color fades rapidly. After 15 minutes, the reaction mixture was poured into water and extracted three times with ethyl acetate. The organic layers were dried over magnesium sulfate and the product purified by flash chromatography on silica gel using 5% methanol in dichloromethane as eluent. 0.2052 g (58%) of an off-white solid product was obtained.

CS 273 199 B2

IR (KBr: 3440, 2930, 1772 (br), 1623 cm ^1st

¹ H-NMR (CDCl 3): δ 12 (ba, 1H), 7.79 (bs, 1H), 7.64 (bs, 1H),

6.73 (β, 1H), 6.64 (s, 1H), 6.26 (β, 1H), 6.24 (a, 1H), 5.73 (m, 2K),

4.75 (d, 0-3.4 Hz, 1H), 4.76-4.43 (m, 4H), 4.31 (d, 0-7.5 Hz, 1H),

4.24 - 4.15 (m, 2H), 4.02 - 3.95 (m, 2H), 3.43 - 3.33 (m, 2H), 3.20 - 3.05 (m, 4H) 1.86-2.78 (m, 1H), 1.16 (d, 0-5.0 Hz, 3H).

Mass spectrum (FAB) m / e 734 (M + H) ⁺ .

Example 5

3 ^from -Demethoxy-5 ^, -demethyl-3-aminoetopoid (IV, B-IVc)

To a solution of 0.205 g (0.273 mmol) of the stoposide dihydroxytriohloroethylcarbamate of Example 4 in 5 ml of tetrahydrofuran, 0.5 ml of water and 0.5 ml of glacial acetic acid is added, with stirring at room temperature, 0.40 g (6.11 mmol) of activated zinc dust. The reaction mixture was suspended in an ultrasonic apparatus for 90 minutes. Thin layer chromatography of the reaction mixture (5% methanol in dichloromethane) showed a product with a lower R, than the starting material. The reaction mixture was poured into water and extracted four times with ethyl acetate. The combined organic extracts were dried over sodium sulfate, concentrated and the residue purified by flash chromatography on silica gel using 5% methanol in dichloromethane as eluent. Yield: 29 mg (19%) of a dark metallic gray solid.

IR (KBr): 3434, 2923, 1767, 1734, 1486 cm ^1st

@ ¹ H-NMR (CDCl3, .delta.):

7.75 (bs, 1H), 6.61 (a, 1H), 6.26 (a, 1H), 5.76 (s, 1H), 5.71 (m, 2H),

5.67 (e, 1H), 4.70 '(d, 0 · 3.2 Hz, 1H), 4.53 (m, 1H), 4.45 (bs, 1H),

4.28 (d, O = 7.7 Hz, 1H), 4.22-4.13 (m, 2H), 3.996 (m, 2H), 3.50 (bm,

1H), 3.34 (m, 2H), 3.15-3.0 (m, 3H), 2.80-2.75 (m, 1H), 1.15 (d, D 4.9 Hz, 3H).

FAB-MS: m / e = 560 (M + H) &lt; + & gt ^; .

Claims (3)

A process for the preparation of nitrogen-containing epipodophyllotoxin glucoside derivatives of the general formula IV B ty CS 273 199 B2 in which B is a radical selected from the group consisting of radicals of formulas IVe, IVb and IVc wherein R ¹¹ OH -COR ¹⁰ or II O (IVb). (IVc) represents a radical -CH, -CR Μ II Wherein R ¹⁰ is an alkyl group of 1 to 5 carbon atoms optionally substituted with 1 to 3 halogen atoms or a phenyl group. -CNHR II Wherein the compound of formula (VI) is a compound of formula (VI) Wherein R ¹¹ is as defined above, oxidizes to form a product of formula XV wherein B is a radical of formula IVa, which then optionally undergoes reduction and cleavage of the acyl group R ¹¹ , 2. A process according to claim 1 wherein the oxidation of the compound of formula (VI) is carried out by treatment with sodium iodate in the presence of acetonitrile and water or with sodium nitrite in the presence of acetic acid and tetrahydrofuran. 3. The process of claim 1 wherein the reduction of the intermediate of formula (IV) in which B is a radical of formula (IVa) is carried out by catalytic hydrogenation or by treatment with sodium metabisulphite.