CH619967A5 - Process for the preparation of nitrosourea derivatives - Google Patents

Description

The present invention relates to a process for the preparation of new nitrosourea derivatives which have a high activity against leukemia and tumors and a low toxicity and are therefore very useful for the therapeutic treatment of leukemia and tumors.

Numerous compounds have already been proposed which have an inhibitory action against leukemia and tumors, but which were unsatisfactory in terms of the level of activity and / or toxicity.

It has now been found that certain new nitrosourea derivatives, as defined below, have a surprisingly high inhibitory activity against leukemia and tumors, while their toxicity is very low, as has been demonstrated in in vivo studies.

The aim of the present invention is therefore to obtain new nitrosourea derivatives of the formula in which A1 is the β-D ~ glucopyranosyl group and R1 is an alkyl group with 2 to 4 carbon atoms or a halogen-substituted alkyl group with 1 to 4 carbon atoms.

Typical examples of compounds of the formula Ia are: 1-ethyl-3- (β-D-glucopyranosyl) -1 -nitrosourea; I- (β-D-glucopyranosyl) -3-ni (roso-3-n-propylurea;

1 "1-n-ButyI-3- (ß-D-glucopyranosyl) -1 -nitrosourea, and 1 - (2-chloroethyl) -3- (ß-D-glucopyranosyl) -1 -nitrosourea.

The second group of preferred compounds of the formula I is represented by the formula Ib:

0

A

■ nhcn

NO

• R

(Ib)

0

a-

• no nhcn

] ■

(I)

- "I in which A2 represents a mannopyranosyl group or a galactopyranosyl group and R2 represents an alkyl group having 1 to 4 carbon atoms or a halogen-substituted alkyl group having 1 to 4 carbon atoms.

Typical examples of compounds of formula Ib include ":

1- (2-chloroethyl) -3- (ß-D-mannopyranosyl) -1-nitrosourea, and

1 - (2-chloroethyl) -3- (ß-D-galactopyranosyl) -1 -nitrosourea. The third group of preferred compounds of the formula I ■ i "is represented by the formula Ic:

0

R

in which A represents a glycosyl group, a monovalent residue of methyl glucoside from which the 2-hydroxy group has been removed, a monovalent residue of alditol from which the 2-hydroxy group has been removed, represents an N-substituted carbamoyloxyalkyl group or a hydroxy-substituted cyclohexyl group if n is 1, or A is a tetravalent radical of ribostamycin from which the four amino groups have been removed when n is 4 and R is a lower alkyl group or a halogen-substituted lower alkyl group, where R is not a methyl group when A represents the glucosyl group.

When A is a glucosyl group, it can be derived not only from a monosaccharide but also from a disaccharide and can be in the form of a deoxy derivative. Likewise, if A is a monovalent residue of methyl glucoside, it can be in the form of a deoxy derivative.

A first group of preferred compounds of the formula Ia corresponds to the new nitrosourea derivatives of the formula I

, s a3-nhcn;

• NO

'R

(Ic)

I in which A1 represents a glycosyl group or a hydroxy-substituted cyclohexyl group and R2 represents an alkyl group with 1 to 4 carbon atoms or a halogen-substituted alkyl group with 1 to 4 carbon atoms.

In the compounds of formula Ic, the glycosyl group z. B. the arabinosyl, xylosyl, lyxosyl, talosyl, idosyl, gulosyl, altrosyl or allosyl group, and the hydroxy-substituted cyclohexyl group can be e.g. B. be a monohydr-oxycyclohexyl group, such as the 2-, 3- or 4-hydroxycyclo-hexyl group, a dihydroxycyclohexyl group, such as the 2,3-, i 2,4-, 2,5-, 2,6-, 3, 4- or 3,5-dihydroxycyclohexyl group, a trihydroxycyclohexyl group such as the 2,3,4-, 2,3,5-, 2,3,6-, 3,4,5- or 3,4,6-trihydroxycyclohexyl group or a tetrahydroxycyclohexyl group such as the 2,3,4,5-, 2,3,4,6-, or 2,3,5,6-tetrahydroxycycIohexyIgruppe.

Typical examples of compounds of formula Ic include:

1-methyl-l-nitroso-3- (β-D-xylopyranosyI) urea; 1 - (2-chloro I) -1 -nitroso-3 ~ (ß-D-xyIopyranosyl) urea;

619 967

6

1 - (1,3 / 2N-dihydroxycyclohexyl) -3-methyl-3-nitrosourea, and

1 - (2-ChloroethyI) -3- (1,3 / 2N-dihydroxycycIohexyI) -l -nitrosourea.

The fourth group of preferred compounds of the formula I is represented by the formula Id:

0

• nh

/

"T \

ch,

(Id)

in which A5 represents the pentahydroxycyclohexyl group and R2 represents an alkyl group with 1 to 4 carbon atoms or a halogen-substituted alkyl group with 1 to 4 carbon atoms.

A typical example of compounds of the formula Ie is: 1 - (2-chloroethyl) -1-nitroso-3- (1 N, 3,5 / 2,4,6-pentahydroxycy-cIohexyl) urea.

The sixth group of preferred compounds of the formula I corresponds to the formula If:

0

■ NHCN,

'NO

in which A4 the mannopyranosyl group, the methyl-2,6-dideoxy-glucopyranoside residue of the formula

OCH.

or the methyl-2-deoxy-6-0-mesyl-β-D-glucopyranoside residue of the formula

VR

(If)

in which A6 represents a glycosyl group derived from Biose and R: -'i 'represents an alkyl group having 1 to 4 carbon atoms or a halogen-substituted alkyl group having 1 to 4 carbon atoms.

As a glycosyl group derived from Biose, the maltosyl, lactosyl, sucrosyl, cellobiosyl, trehalosyl, gentiobiosyl, melibiosyl, turanosyl, sophorosyl or isosu-crosyl group can be mentioned.

Typical examples of compounds of the formula If um 1 - (β-D-maltosyl) -3-methyl-3-nitrosourea; ìn 1 - (2-chloroethyl) -3- (ß-D-maltosyl) -1 -nitrosourea; 1 - (ß-D-lactosyl) -3-methyl-3-nitrosourea, and 1 - (2-chloroethyl) -3- (ß-D-lactosyl) -1 -nitrosourea.

The seventh group of preferred compounds of the formula I corresponds to the formula Ig:

CH2OMS

0I \ OH H / H

OCH.

0

A 'NHC

NO

(ig)

R

in which A7 is a monovalent alditol radical from which the 2-hydroxy group has been removed and which has the formula:

in which Ms represents the mesyl group.

Typical examples of Formula Id include: 1 - (β-D-mannopyranosyl) -3-methyl-3-nitrosourea; Methyl-2,6-dideoxy-2- (N'-methyl-N'-nitrosoureido) -ß-D-glu-copyranoside, and

Methyl-2-deoxy-6-0-mesyl-2- (N'-methyl-N'-nitrosoureido) -ß-D-glucopyranoside.

The fifth group of preferred compounds of the formula I corresponds to the formula Ie:

CH2OH

CH-

(CHOH)

n ch2oh

0

• NHCN

, N0

'r

(Ie)

in which n represents zero or an integer from 1 to 3 and R2 represents a halogen-substituted alkyl group having 1 to 4 carbon atoms.

The monovalent alditol residue as A7 can be derived from tritol, tetritol, pentitol or hexitol.

Typical examples of compounds of the formula Ig include:

N-carbamoyl-N '- (2-chloroethyl) -N'-nitroso-D-glucosaminol, and

7

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N-Carbamoyl-N '- (2-chloroethyl) -N'-nitroso-D-galactosaminol.

The eighth group of preferred compounds of the formula I corresponds to the formula Ih:

j »o

-NHCNI

'R

(ih)

in which As is an N-substituted carbamoyloxylakyl group of the formula

0

R '

■ 0

: only in which R- 'is an alkylene group with 2 to 5 carbon atoms and R4 is an alkyl group with 1 to 4 carbon atoms or a halogen-substituted alkyl group with 1 to 4 carbon atoms, and R2 is an alkyl group with 1 to 4 carbon atoms or a halogen-substituted alkyl group with 1 to Represents 4 carbon atoms.

As the alkylene group R3, the ethylene, trimethylene, propylene, tetramethylene, 1,2-dimethylethylene, pentamethylene, 1-methyltetramethylene, 2-methyltetramethylene, 1,2-dimethyltrimethylene and 1,3- DimethyItrimethylengruppe be called.

Typical examples of compounds of the formula Ih include:

(N-carbamoyl-N'-methyl-N'-nitroso) - (O-carbamoyl-methyi) -iithanolamine, and

[N-CarbamoyI-N '- (2-chloroethyl) -N'-nitroso] - (0-carbamoyl-N'-2-chloroethyl) -; ithanolamine.

The ninth group of preferred compounds of formula 1 is represented by formula Ii:

0

-NHCN

(Ii)

in which A1 'represents the fi-deoxy-glucopyranosyl group and R2 represents an i-alkyl group with 1 to 4 carbon atoms or a halogen-substituted alkyl group with 1 to 4 carbon atoms.

Typical examples of compounds of the formula Ii are: 1- (6-deoxy-β-D-glucopyranosyl) -3-methyl-3-nitrosourea, »and

1 - (2-chloroethyl) -3- (6-deoxy-ß-D-glucopyranosyI) -1 -nitrosourea.

The tenth group of preferred groups of the formula I s is represented by the formula Ij:

0

A10 1-NHCN

NO

]

(Ij)

> R

in which A1 "is a tetravalent radical of ribostamycin of the formula

619 967

8th

and R2 represents an alkyl group having 1 to 4 carbon atoms or a halogen-substituted alkyl group having 1 to 4 carbon atoms.

Typical examples of compounds of formula Ij include:

Tetra-N- (N'-methyl-N'-nitroso) carbamoyl ribostamycin, and

Tetra-N- [N '- (2-chloroethyI) -N'-nitroso] -carbamoyl-ribosta-

mycin.

The new nitrosourea derivatives of formula I have proven to be highly active in inhibiting leukemia and tumors with low toxicity.

These compounds can therefore be processed into useful pharmaceutical preparations which contain a nitrosourea derivative of the formula I together with a pharmaceutical carrier or diluent.

The pharmaceutical preparation can be in any known form. The nature of the preparation and the pharmaceutical carrier or diluent used depends, of course, on the intended route of administration, e.g. B. orally or parenterally. In general, the preparation may be in a form suitable for injection or oral administration, e.g. B. in the form of ampoules, capsules, tablets, powder, granules and the like.

For the treatment of leukemia and tumors in humans and animals, the patients are administered a therapeutically effective amount of a nitrosourea compound of the formula I at suitable intervals. The amount of the nitrosourea derivative administered depends on the compound used in each case, the composition of the preparation, the mode of use and the mode of administration and other factors. Thus, numerous factors that can modify the effect of the drug, such as age, body weight, gender, diet, time of administration, route of administration, speed of metabolism or excretion, drug combination, sensitivities and degree or state of the disease, will be taken into account by the person skilled in the art . The optimum administration dose under the given conditions can easily be determined by the person skilled in the art using conventional examination methods.

The chemotherapeutic efficacy of a large number of the nitrosourea derivatives of the formula I against leukemia L1210 in mice was investigated using the following method:

Examined compounds compound name

No.

1

1 - (2-chloroethyl) -3- (ß-D-glucopyranosyl) -1 -

nitrosourea

2nd

1 - (2-chloroethyl) -3 - (ß-D-mannopy ranosyl) -1 -

nitrosourea

3rd

1 - (2-chloroethyl) -1 -nitroso-3- (ß-D-xylopyra-

nosyl) urea

4th

1 - (2-chloroethyl) -3- (1,3 / 2N-dihydroxycyclo-

hexyl) -1 nitrosourea

5

l- (2-chloroethyl) -l-nitroso-3- (lN, 3,5 / 2,4,6-

pentahydroxycyclohexyl) urea

6

Methyl 2,6-dideoxy-2- (N'-methyl-N'-nitro-

soureido) -ß-D-glucopyranoside

7

1 - (2-chloroethyl) -3- (ß-D-galactopyranosyI) -1 -

nitrosourea

8th

l- (2-chloroethyl) -3- (ß-D-lactosyl) -l-nitroso

urea

9

N-carbamoyl-N '- (2-chloroethyl) -N'-nitroso-D-

glucosaminol

10th

"Daunomycin" (comparative substance 1)

11 l- (β-D-glucopyranosyl) -3-methyl-3-nitroso-urea (comparative substance 2)

12 N-carbamoyl-N'-methyl-N'-nitroso-D-glucosaminol (comparative substance 3)

Laboratory animals

BDF, mice (about 6 weeks old; body weight: 22 ± 1 g); each study group consisted of 2 male mice.

BDFr mice (about 7 weeks old; body weight:

21 ± 2 g); each study group consisted of 3 male mice.

BDF mice (about 7 weeks old; body weight:

22 ± 2 g); each study group consisted of 2 male mice.

BDF mice (about 6 weeks old; body weight 21 ± 2 g); each study group consisted of 3 male mice.

BDFr mice (about 6 weeks old; body weight:

20 ± 1 g); each study group consisted of 3 female mice.

BDFr mice (about 6 weeks old; body weight:

21 dt 1 g); each study group consisted of 3 male mice.

or lines which were inoculated for the examinations ml / ml / ml /

The compound to be examined was dissolved in a physiological saline solution and administered intraperitoneally in a volume of 0.1 ml / mouse. It was administered once a day for three days, the first 24 hours after the intraperitoneal vaccination of the L1210 cells. The effect of the compound to be examined was determined on the basis of the survived days of the mice, the percentage increase in the lifespan and the anatomical findings, including the volume of the ascites.

B The same procedure as in A, but administration of the drug was continued for 5 days. 5 „C The same procedure as under A, but the compound to be examined was suspended in 0.5% CMC solution and the administration of the drug was continued for 5 days.

D The same procedure as under A, but the compound to be examined was dissolved in distilled water for injections.

The results of the tests are summarized in the table. The percentage increase in lifespan f (| (ILS) was calculated as follows:

Percentage increase in lifespan (ILS) = ——— X100

T: The average survived days of the treated animals.

C: The average survived days of the untreated control animals.

m

B

C.

is

D

T

A cells from Leukemia L1210 (2.1 X 106 cells / 0.05 111 mouse) were inoculated intraperitoneally. B cells from Leukemia L1210 (1.9 X 10f 'cells / 0.05

Mouse) were inoculated intraperitoneally. C cells from Leukemia L1210 (1.2 X 106 cells / 0.05 mouse) were inoculated intraperitoneally.

IS

Test procedure

9

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table

Connect

Ver

Do

test

Dose-

Survived days

ILS

Ascites volume of such mg mg / kg

Treated / control

%

Treated / control

No.

animals

drive

10th

95.9 /

9.3

931.1

0 / 4.7

8th

95.9 /

9.3

931.1

0 / 4.7

6.25

> 40.1 /

9.3

> 331.2

0 / 4.7

5

15.37

9.3

64.5

1.5 / 4.7

1

A

A

A

4th

40.3 /

9.3

333.3

0 / 4.7

2nd

15.3 /

9.3

64.5

2.5 / 4.7

1

12.9 /

9.3

38.7

5.0 / 4.7

0.5

12.5 /

9.3

34.4

4.2 / 4.7

0.2

12.0 /

9.3

29.0

4.2 / 4.7

8th

18.3 /

9

103.3

0 / 2.8

4th

15.0 /

9

66.7

1.2 / 2.8

2nd

D

A

A

2nd

14.7 /

9

63.0

1.2 / 2.8

1

13.7 /

9

51.9

2.3 / 2.8

0.5

13.0 /

9

44.4

3.0 / 2.8

16

9.0 / 11.0

-18.2

0 / 3.0

8th

> 60.0 / 11.0

> 445.5

0 / 3.0

3rd

D

A

A

4th

> 60.0 / 11.0

> 445.5

0 / 3.0

2nd

> 24.0 / 11.0

> 118.2

0 / 3.0

1

17.0 / 11.0

54.5

0 / 3.0

16

13.0 / H), 3

26.2

0 / 2.7

4th

B

B

A

8th

> 59.3 / 10.3

> 475.7

0 / 2.7

4th

> 60.0 / 10.3

> 482.5

0 / 2.7

2nd

22.0 / 10.3

113.6

0.8 / 2.7

30th

11.6 /

9.7

19.6

0 / 1.5

20th

> 61.1 /

9.7

> 529.9

0 / 1.5

10th

15.5 /

9.7

59.8

0 / 1.5

5

A

A

A

8th

16.0 /

9.7

64.9

2.5 / 1.5

6

14.0 /

9.7

44.3

2.0 / 1.5

4th

13.0 /

9.7

34.0

2.8 / 1.5

2nd

12.5 /

9.7

28.9

4.0 / 1.5

1

12.0 /

9.7

23.7 -

4.5 / 1.6

6

C.

C.

B

300

13.0 /

8.5

52.9

0.3 / 4.0

200

15.1 /

8.5

77.6

2.0 / 4.0

10th

18.7 /

9.3

101.1

0 / 3.7

8th

> 47.0 /

9.3

> 405.4

0 / 3.7

7

B

B

A

4th

> 47.0 /

9.3

> 405.4

0 / 3.7

2nd

14.7 /

9.3

58.1

0.8 / 3.7

1

18.5 /

9.3

98.9

4.6 / 3.7

10th

> 47.0 /

9.7

> 384.5

0 / 4.2

8th

E

A

A

5

14.3 /

9.7

47.4

1.0 / 4.2

2.5

12.5 /

9.7

28.9

3.3 / 4.2

9

F

B

A

16

> 53.7 / 10.7

> 401.8

0 / 2.5

8th

18.0 / 10.7

68.2

0 / 2.5

£

2nd

13.6 /

8.5

60.0

10th

C.

C.

C.

1

12.6 /

8.5

48.3

0.5

12.8 /

8.5

50.6

0.25

10.2 /

8.5

20.0

200

16.6 /

8.0

107.5

1.4 / 3.1

11

D

A

A

150

14.1 /

8.0

83.4

1.9 / 3.1

100

13.9 /

8.0

80.7

4.6 / 3.1

50

12.0 /

8.0

55.6

2.8 / 3.1

400

13.4 / 10.7

25.0

3.4 / 2.5

12

F

B

D

200

15.0 / 10.7

40.0

4.0 / 2.5

100

13.9 / 10.7

30.0

3.0 / 2.5

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10th

It is clearly evident from the test results from the table that the new nitrosourea derivatives of the formula I result in a considerable increase in the lifespan in the therapeutic treatment of leukemia in comparison with «Dau-nomycin», which was used clinically for the treatment of leukemia, and that they had little or no accumulation of ascites. The new nitrosourea derivatives are also notable for their low toxicity, which makes them ideal for practical use in the treatment of leukemia and tumor diseases. For example, a typical nitro-urea derivative, namely 1 - (2-chloroethyl) -3- (ß-D-glucopy-ranosyI) -l-nitrosourea, gave an LD5l) value of approximately 25 mg / kg when administered intraperitoneally to BDFr mice.

The new nitrosourea derivatives of the formula I can be obtained by nitrosation of the corresponding urea derivatives.

The process according to the invention for the preparation of nitrosourea derivatives of the formula:

{

0

"NO '

1

NHCN I n

\ T

(i)

in which A is a glycosyl group, a monovalent residue of a methyl glucoside from which the 2-hydroxy group has been removed, a monovalent residue of alditol from which the 2-hydroxy group has been removed, an N-substituted carbamoyloxyalkyl group or a hydroxy-substituted cyclohexyl group if n is 1, or a tetravalent radical of ribostamycin from which the four amino groups have been removed when n is 4 and R represents a lower alkyl group or a halogen-substituted lower alkyl group, wherein R is not a methyl group when A represents the glucosyl group, thereby characterized that you have a compound of formula

0

r "l

J- NHCNHRJ n

Solvents are recrystallized to obtain the desired product in high purity.

The compounds of the formula II which are used as starting material in the process according to the invention are likewise new compounds and can be prepared by using a compound of the formula

A-NH2 (III)

in which A means the same as above, with a lower alkyl or a halogen-substituted lower alkyl isocyanate of the formula

R-N = C = O (IV)

in which R means the same as above. Before the reaction, the alcoholic hydroxyl groups present in group A can, if necessary, be replaced by one of the many known OH protecting groups, e.g. As acetyl, are protected.

A large number of isocyanates can be used as compounds of the formula IV. Typical examples of isocyanates of the formula IV are methyl, ethyl, n-propyl, i-propyl, n-butyl, i -Butyl-, sec-butyl-, tert-butyl-, chloromethyl-, ß-chloroethyl-, y-chloropropyl-, ß-chloropropyl-, ô-chlorobutyi and ß-bromoethyl isocyanate.

Thus, the starting compounds of formula II which are used for the preparation of compounds of formula Ia, Ib, Ic in which A3 represents a glycosyl group, Id in which A4 is the mannopyranosyl group and If can be prepared in the same way by Acetyl-glycopyranosylamine with i "an isocyanate of formula IV in an inert organic solvent, such as. B. methanol, and the reaction product is deacetylated. As a typical example of this preparation, a scheme for the preparation of 1 - (β-D-mannopyrano-syl) -3-methylurea is explained below.

s p

Penta-O-acetyl-D-mannopyranose of the formula

ÇH20AC

a n \ OAc AcO. _. AcO \ i i / OAc

(Ii)

in which A, R and n have the meaning given above, reacted with a nitrosating agent.

In this process, the nitrosation reaction can be carried out in a manner known per se by using an alkali metal nitrite, nitrogen trioxide, nitrous oxide and the like as the nitrosating agent. Preferably the alkali metal nitrite is sodium or potassium nitrite. The reaction can usually be carried out at a temperature of about -10 to 30 ° C and preferably under acidic conditions, e.g. B. a pH of about 1 to 3. Under these conditions, the reaction time is advantageously about 1 to 12 hours. Water, a lower fatty acid such as formic acid, acetic acid or propionic acid or a mixture thereof can advantageously be used as the reaction medium.

After the nitrosation reaction has ended, the reaction product can, if desired, be purified in a conventional manner, e.g. B. using a cation exchange resin, then lyophilized, and then from a suitable su in which Ac represents the acetyl group, which can be prepared in a known manner (PA Levene, et al., J. Biol. Chem., 90 "89 (1931 ) is used as the starting material and treated with HBr and NaN3 in CH3CN and then hydrogenated in the presence of a Raney nickel catalyst to produce the compound of the following formula:

ÇH20Ac

AcO

11

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in which Ac has the same meaning as above, whereupon this compound is reacted with the compound CH3NC = 0 to give the compound of the formula

CH2OAC

AcO

OAc AcO

nhconhch.

ch2oh, in which Ac again has the same meaning as above, whereupon this compound is deacetylated to give the desired compound of the formula which can be prepared in a known manner (A. Neuberger et al., J. Chem. Soc., 122 (1937) is used as the starting material and is reacted with tosyl chloride or mesyl chloride to produce the compound of the formula ch2oh nhcnhch.

CH2OTs (or Ms)

och

NHC00CH

to surrender.

The starting compounds of the formula II, which are used for the preparation of the compounds of the formula Id, in which A4 the methyl-2,6-dideoxy-β-D-glucopyrano-sidrest of the formula och.

in which Ts is the tosyl group and Ms is the mesyl group, whereupon this compound is reacted with sodium iodide to form the compound of the formula, can be prepared, for example, in the following manner:

The compound of the formula

OCH.

nhcooch which is then hydrogenated in the presence of a Raney nickel catalyst and then reacted with an isocyanate of the formula IV to give the desired compound of the formula

619967

12

och

nhcnhr in which R has the same meaning as above.

The compounds of formula II, which is used for the preparation of the compounds of formula Id, in which A4 the methyl-2-deoxy-6-0-mesyI-ß-D-glucoside residue of the formula

CH2OMs och.

means in which Ms represents the mesyl group, z. B. can be obtained in the following way:

The compound of the formula ch2oh och,

ho oh och

in which Ms represents the mesyl group and R has the same meaning as above.

The starting compounds of the formula II, which are used for the preparation of compounds of the formula Ic in which A3 represents a hydroxy-substituted cyclohexyl group, can be obtained in a simple manner by reacting a hydroxy-substituted cyclohexylamine with an isocyanate of the formula IV. The hydroxy-substituted cyclohexylamine serving as the starting material can be prepared in a known manner (T. Suami and S. Ogawa, Bull. Chem. Soc. Japan, 37, 194 (1964).

The starting compounds of formula II, which are used for the preparation of compounds of formula Ie can be prepared in the same manner as the latter compounds, namely by reacting an inosamine with an isocyanate of formula IV in an inert organic solvent, such as. B. methanol. Various isomers can be used as the inosamine, including the scyllo, allo-, myo-, muco-, cis-, neo-, chiro- and epi-i inosamines. On the other hand, deoxyinosamines, such as mono-, di-, tri- and tetradeoxyinosamines, can also be used for the reaction with an isocyanate of the formula IV.

The starting compounds of the formula II, which are used for the preparation of compounds of the formula Ig, can be obtained by reacting an aminoalditol of the formula

»? Hz0H

in which R has the meaning given above, which can be prepared in a known manner (T. Suami, et al., Bull. Chem. Soc. Japan, 43, 3013 (1970) is used as the starting material and mesylated with mesyl chloride to form the Compound of the formula ch20mS

ch-nh.

i '

(choh)

n ch2oh with an isocyanate of the formula IV in an inert solvent, such as water, methanol, ethanol or acetone. As typical examples of amino alditols, aminotritols, aminotetritols, aminopentitols and amino hexitols can be mentioned. The most typical aminohexitols are glucosaminol and galactosaminol.

The starting compounds of formula II, which are used for the preparation of compounds of formula Ih, can, for. B. by reacting an alkanolamine of the formula

H2N-R3-OH

in which R3 represents an alkylene group with 2 to 5 carbon atoms, with an isocyanate of the formula IV in an inert organic solvent, such as methanol, ethanol or acetone.

The starting compounds of formula II, which are used for the preparation of compounds of formula Ii, can, for. B. can be obtained using the following known method, which comprises the following stages:

nhconhr

13

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ch, çh3

i 0 1 o

/ i \ nhconhch3 / \ nhconch3

i / 1 '

oh 1 oh l5.

The starting compounds of formula II, which are used for the preparation of compounds of formula Ij, can be easily obtained by reacting ribosta-mycin with an isocyanate of formula IV in an inert organic solvent, e.g. B. methanol.

The new nitrosourea derivatives produced according to the invention are also useful as intermediates for the production of certain other chemicals which can be used for pharmaceutical purposes.

The new nitrosourea derivatives of the formula I can be converted into the corresponding acetyl derivatives in a simple manner by treating them with a mixture of pyridine and acetic anhydride.

"Amberlite IR-120" (H + form) treated to remove the sodium ion from it. The reaction mixture was then freeze-dried, whereby 1.0 g of the compound mentioned in the title in the form of an amorphous substance of pale yellow

Color was obtained, [a] ^ - 2 ° (c = 1.0, H20). These

Substance gave a single spot at Rf 0.82 in thin layer chromatography on silica gel using chloroform-methanol (3: 2 by volume ratio) as an emulsifier.

N.M.R. spectrum (in D20):

x4.74 (d, IH, J 9 Hz, H-l)

8.99 (t 3H, J 7 H7, C-CH,).

Example 1 "u

(a) 1-Ethyl-3- (ß-D-glucopyranosyl) -I-nitrosourea l-ethyl-3- (ß-D-glucopyranosyI) urea (930 mg) was added to a mixture of water (4.3 ml ) and acetic acid (4.3 ml) and 370 mg (1.5 mol equivalents) of sodium nitrite are added to this solution in small portions with stirring. The mixture was stirred at room temperature overnight to complete the nitrosation. The reaction mixture was then treated with 4 ml of a cation exchange resin,

(b) Ì-Àthyl-3- (tetra-O-acetyl-ß-D-gtucopyranosyl) -1-nitrosourea

1-Ethyl-3- (ß-D-gliicopyranosyI) -l -nitrosourea (1.0 g) was dissolved in a mixture of pyridine (6 ml) and acetic anhydride (6 ml) and the resulting solution was left to stand at room temperature overnight to complete the acetylation. The reaction mixture was then evaporated to dryness under reduced pressure and the residue was crystallized from ethanol. The desired connection

619 967

14

was obtained in a yield of 1.0 g (67%). Melting point: 116 to 118 ° C.

[a]

24 D

-8 ° (c = 1.0, chloroform).

Elemental analysis:

calculated for C17H25N30n: C45.63, H 5.64, found: C 45.89, H 5.60,

N 9.39% N 9.34%

Example 2

(a) I- (ß-D-glucopyranosyl) -3-nitroso-3-n-propylurea l- (ß-D-glucopyranosyl) -3-n-propylurea (1.50 g) was added to a mixture of water ( 10 ml) and acetic acid (2 ml) and the resulting solution was mixed with 560 mg (1.5 mol equivalents) of sodium nitrite at room temperature with stirring to effect the nitrosation. The reaction mixture was treated with “Amberlite IR-120” (H + form) in order to remove the sodium ions and then evaporated to dryness at a temperature below 35 ° C. and under reduced pressure. Crystallization of the residue from n-propanol gave 1.35 g of the desired compound. Yield: 80%. Melting point: 112 ° C (decomposes under foaming

men). [a] ^ -g ° (c = 1.0, H20).

Elemental analysis: calculated i found:

calculated for C10H19N3O7:

C 40.95, C 40.77,

H 6.54, H 6.69,

N 14.33% N 13.93%

IR spectrum:

3300 (attributable to OH)

1700 (attributable to CO)

1530 (attributable to NH) and 1490 cm "" 1 (attributable to N-NO).

N.M.R. spectrum: (in D20):

t4.77 (d, lH, J9Hz, H-l),

9.22 (t, 3H, J 7 Hz, C-CH3).

(b) / - (Tetra-0-acetyl-ß-D-glucopyranosyl) -3-nitroso-3-n-pro-pylurea l- (ß-D-glucopyranosyI) -3-nitroso-3-n-propylurea in the same way as in Example 1 (b) to the desired

(b) 1 - "- Butyl-3- (tetra-0-acetyl ~ β-D-glucopyranosyl) -l-nitrosourea

1-n-Butyl-3- (β-D-g! Ucopyranosyl) -1 -nitrosourea was obtained in the same manner as in Example 1 (b)

Acetylated compound. Melting point: 127 to 128 ° C. [a] ^

- 12 ° (c = 1.0, chloroform).

Example 4

in (a) 1 - (2-chloroethyl) -3- (ß-D-glueopyranosyl) -l-nitrosourea

A solution of 0.60 g of 1 - (2-chloroethyl) -3- (ß-D-glucopy-ranosyl) urea in 15 ml of 99% formic acid was mixed with 282 mg of sodium nitrite with ice cooling and stirring, l <• Das The mixture was stirred under ice-cooling for 1 hour to carry out the nitrosation. After the addition of 15 ml of cold water, the reaction mixture was ice-cooled for 30 minutes and then treated with 5 ml of “Amberlite IR-120” (H + form) in order to remove the sodium cations. The reaction mixture treated in this way was evaporated to dryness at a temperature below 30 ° C. by distilling off the solvent under reduced pressure. The residue was taken up in a small volume of n-propanol and the solution mixed with a volume of petroleum ether to form a precipitate. The precipitate that had settled was filtered off and then dried in a desiccator under vacuum, giving 0.65 g of the desired compound as an amorphous solid of a pale yellow color. Yield:

92%. Melting point: 85 ° C (decomposed), [a] p -14 °

(c = 0.5, methanol).

Elemental analysis:

calculated for: Cc, HlfiN3C107:

C 34.46, H 5.14, found: C 34.36, H 5.09,

N 13.40, N 13.20,

Cl 11.30% Cl 11.33%

th compound acetylated. Melting point: 111 ° C. [a] ^ (c = 0.5, chloroform).

• 10 °

Example 3

(n) 1-n-Butyl-3- (β-D-glucopyranosyl) -1 -nitrosourea In-butyl-3- (β-D-glucopyranosyl) urea (0.50 g) was mixed in a mixture of water ( 2 ml) and acetic acid (2 ml)

dissolved and the solution obtained was mixed with sodium nitrite (186 mg, 1.5 molar equivalents). The nitrosation was carried out in the same manner as described in Example 1 (a). The reaction mixture was treated with “Amberlite IR-120” (H + form) to remove the sodium cations and

then freeze-dried, whereby 0.5 g of the desired compound was obtained as a glass-like substance of pale yellow color, [a] ^ -1.5 ° (= 1.0, H20). This substance

<

gave a single spot at Rf 0.51 in a thin layer chromatogram on silica gel using chloroform-methanol (4: 1 by volume ratio) as the eluent.

I.

N.M.R. spectrum (in D20):

t4.74 (d, IH, J 7.5 Hz, H-l),

9.13 (t, 3H, J 6.5 Hz, C-CH,).

(b) 1 - (2-Chloroethyl) -3- (tetra-0-acetyl-ß-D-glucopyranosyl) -l -nitrosourea

1 - (2-Chloroethyl) -3- (ß-D-glucopyranosyl) -l -nitrosourea was treated with acetic anhydride in the same manner as described in Example 1 (a), to give the desired compound. Melting point: 103 to 105 ° C.

[a] p -2 ° (c = 0.5, methanol).

Example 5

1 - (2-Chloroethyl) -3- (ß-D-mannopyranosyl) -I -nitrosourea l- (2-chloroethyl) -3- (ß-D-mannopyranosyl) -urea (500 mg) was dissolved in 10 ml 99% Formic acid dissolved and the solution obtained mixed with 250 mg of sodium nitrite in an ice bath with stirring, the sodium nitrite being added to the solution in small portions. The mixture was stirred for 1 hour and then 10 ml of cold water was added, followed by stirring for another 30 minutes to complete the nitrosation. The reaction solution was then treated with 20 ml of “Amberlite IR 120” (H + form) and then shaken for 20 minutes to remove the sodium cation. After the cation exchange resin was filtered off from the reaction mixture, the filtrate was evaporated to dryness by evaporating the solvent under reduced pressure. A crystalline solid was obtained. This solid was digested with ethanol to give the desired compound. Yield: 308 mg (55.9%). Melting point: 98 to 100 ° C (decomposition under foaming), [a] ^ -10 ° (c =, 5, H20).

15

619 967

Elemental analysis:

calculated for CgH16N3C107 (molecular weight: 313.69):

C 34.46, H 5.14, N 13.40, Cl 11.30% found: C35.21, H5.61, N 11.69, Cl 12.07%.

Example fi

(a) 1- (2-chloroethyl) -3- (ß-D-galaetopyranosyl) - / - nitrosourea

150 mg of sodium nitrite were slowly added in an ice bath with stirring to a solution of 300 mg of 1- (2-chloroethyI) -3- (β-D-galactopyranosyl) urea in 2 ml of 99% formic acid. The mixture was stirred under ice cooling for 1 hour to complete the nitrosation. The reaction mixture was then mixed with 50 ml of ethyl ether, an oily substance being eliminated. The ethyl ether phase was decanted from the oily substance. The addition of ethyl ether and the separation of the ethyl ether phase from the oily substance by decantation was repeated three times using 50 ml, 20 ml and 20 ml ether, respectively. The oily substance was then taken up in 30 ml of methyl alcohol and the solution obtained was mixed with 3 ml of “Amberlite IR-120” (H + form). The mixture was filtered to remove the cation exchange resin and the filtrate was concentrated under reduced pressure, whereby crystals of the desired compound were precipitated. Yield: 210 mg (63.6%). Melting point: 145 ° C

(Decomposition under foaming), [a] ^ + 13.0 ° (c = 0.5, H20).

IR spectrum: 1720, 1535 (ureido) and 1495 cm'1 (nitrosamin).

NMR spectrum: Ô 3.52 (t, 2H, J 7 Hz, N-CH2-CH2-C1) Ô 4.28 (t, 2H, J 7 Hz, N-CH2-CH2-C1) ô 6.03 (t, 1H, Jnii = Jl29Hz, Hl) ô 10.57 (d, IH, J 9Hz, NH)

Elemental analysis:

calculated for C9HU, N3C106 (molecular weight: 313.5):

C 34.46, H 5.14, N 13.40, Cl 11.30% found: C34.58, H5.09, N 13.12, Cl 11.22%. ) n

(b) /, (2-chloroethyl) -3- (tetra-0-acetyl-ß-D-galactopyranosyl) - / -nitrosourea

1 - (2-Chloroethyl) -3- (ß-D-galactopyranosyl) -l -nitrosourea (115 mg) was dissolved in 2 ml of anhydrous pyridine and 1 ml of acetic anhydride was added dropwise to this solution while cooling with ice and stirring. The mixture was stirred under ice cooling for 5 hours to complete the acetylation. The reaction mixture was then evaporated to dryness under reduced pressure and the residue was crystallized from water to give 138 mg (78.1%) of the desired compound. Melting point:

69 to 70 ° C. [ex] 19> 5 + 5.0 ° (c = 0.5, chloroform).

D

5 s

Example 7

(a) 1-Methyl-3- (ß-D-xyiopyranosy!) urea

2,3,4-Tri-O-acetyI-ß-D-xylopyranosyl azide (1.03 g; 3.4 mil-limol) was dissolved in ethyl acetate (10 ml) and in the presence of platinum oxide (29 mg) as a catalyst Room temperature "" catalytically hydrogenated for 1 hour, the initial hydrogen pressure being 3.50 kg / cm 2, whereby the azido group was converted into the amino group.

Thin layer chromatography of the reaction product on silica gel using toluene-methyl ethyl ketone (volume ratio 1: 1) as eluent gave no spot at Rf 0.84, which corresponds to the starting material, but a single spot at Rf 0.39.

After the catalyst had been removed by filtration, the reaction mixture was washed with ethyl acetate (10 ml) and the filtrate was combined with the washing liquids and then mixed with methyl isocyanate (0.8 ml, 12.8 millimoles) and the mixture obtained for 1. Stirred for 5 hours under ice cooling and then left overnight at room temperature.

Thin layer chromatography of the reaction product on silica gel using benzene-ethanol (volume in ratio 5: 1) as the eluent gave no spot at Rf 0.62, which corresponds to the starting material, but a main spot at Rf 0.53.

After removal of the solvent by distillation under reduced pressure, the reaction product was crystallized from i * isopropanol, whereby high crystals (979 mg) of melting point 165 to 168 ° C. were obtained. Recrystallization from isopropanol gave l-methyl-3- (2,3,4-tri-0-acetyl-β-D-xylopyranosyl) urea (801 mg) in a pure state.

Yield: 70.7%. Melting point: 170 to 172 ° C. [a] "ÏJ + 43 ° (c = 1.4, chloroform).

560 mg of the compound thus obtained was dissolved in 1.5 ml of methanol, the solution was mixed with 2 ml of 0.1N sodium methoxide, and the resulting mixture was left in a refrigerator overnight to effect deacetylation. The precipitate which separated out was collected by filtration and gave 284 mg of crude crystals. Recrystallization from methanol gave the desired compound (205 mg). Yield: 61.9%. Melting point: 197.5 to 198.0 ° C (below

Decomposition), [ot]

20 _ D

27.3 ° (c = 0.7, H20).

Elemental analysis:

calculated for C7H, 4N205: C 40.77, H 6.84, found: 'C 40.50, H 6.67,

N 13.59% N 13.26%.

(b) / -Methyl-I-nitroso-3- (ß-D-xylopyranosyl) urea l-methyl-3- (ß-D-xyIopyranosyl) urea (418 mg, 2.0

Millimoles) and sodium nitrite (174 mg, 2.5 millimoles) were dissolved in 3 ml of water and 0.5 ml of glacial acetic acid were added to the resulting mixture with stirring and ice cooling and kept under the same conditions for a further 2 hours and then overnight in a refrigerator ditched.

Thin layer chromatography of the reaction product on silica gel using chloroform-methanol (3: 1 by volume) did not give a spot at Rf 0.19, which corresponds to the starting material, but a single spot at Rf 0.60.

After treatment with "Amberlite IR-120" (H + form) in a known manner to remove the sodium cations, followed by distillation under reduced pressure to remove the solvent used, a crystalline residue was obtained. The residue was digested with ethanol to give the desired compound (299 mg). Yield: 62.7%. Melting point: 109 to 110 ° C

(with decomposition), [a] ^ - 26.8 ° (c = 0.6, H20).

IR spectrum (cm1): 1485 (N-NO)

Elemental analysis:

calculated for C7H13N30 (1: C35.74, H 5.57, N 17.87% found: C35.92, H 5.54, N 17.65%.

(c) 1-Methyl-1-nitroso-3- (2,3,4-tri-O-acetyl-β-D-xylopyra-nosyl) urea

1-Methyl-1-nitroso-3- (β-D-xylopyranosyl) urea (110 mg) was dissolved in a mixture of pyridine (1 ml) and acetic anhydride (1 ml) and the resulting mixture was added

619 967

16

Allow to stand at room temperature to effect acetylation. The reaction mixture was then concentrated under reduced pressure to give a crystalline residue, which was then crystallized from ethanol to give 147 mg of the desired compound. Yield; 87.0%.

Melting point: 127 to 127.5 ° C (with decomposition). [«] ^

-25.4 ° (c = 0.7, chloroform).

I.R. spectrum (cm-1): 1495 (N-NO).

m

Example 8

(a) 1- (2-chloroethyl) -3- (β-D-xylopyranosyl) urea 2,3,4-tri-O-acetyl-β-D-xylopyranosyl azide (1.94 g, 6.4 millimoles) was dissolved in ethyl acetate (15 ml) and subjected to catalytic hydrogenation in the presence of platinum oxide (45 mg) as a catalyst at room temperature for 1 hour at an initial hydrogen pressure of 3.50 kg / cm 2 to convert the azide group to the amino group .

Thin layer chromatography of the reaction product on silica gel using toluene-methyl ethyl ketone.> U (volume ratio 1: 1) gave no spot at Rf 0.84, which corresponds to the starting product, but a single spot at Rf 0.39.

After removal of the catalyst by filtration, the reaction mixture was washed with ethyl acetate (10 ml) and: * • the filtrate was combined with the washing liquids, and then 2-chloroethyl isocyanate (1.5 ml, 17.7 millimoles) was added and the resulting mixture was kept for Stirred for 1.5 hours under ice cooling.

Thin layer chromatography of the reaction product on silica gel using toluene-methyl ethyl ketone (volume ratio 1: 1) as the eluent gave a major spot at Rf 0.52 and a minor spot at Rf 0.40.

The reaction mixture was concentrated under reduced pressure to remove the solvent used,

leaving a syrup to which 0.1N sodium methoxide (7 ml) was added, after which the homogeneous mixture thus obtained was left to stand in a refrigerator to effect deacylation. The precipitate formed was collected by filtration to give 1.25 g of crude crystals. Recrystallization from methanol gave the desired compound (768 mg). Yield: 46.9%. Melting point 153 to 155 ° C (with decomposition), [a] ^ -11.0 °

(c = 1.1, H20). 45

Further recrystallization gave a sample of analytical purity. Melting point: 159.5 to 160 ° C (with decomposition). [a] 2p -13.0 ° (c = 0.7, H20).

SO

(b) i- (2-Chloroethyl) -l-nitroso-3- (ß-D-xylopyranosyl) urea l- (2-chloroethyl) -3- (ß-D-xylopyranosyl) urea

(314 mg), 1.2 millimoles) was dissolved in 99% formic acid (3 ml), sodium nitrite (173 mg, 2.5 millimoles) was added to the solution, and the mixture was stirred under ice-cooling for 2 hours.

Thin layer chromatography of the reaction product on silica gel using chloroform-ethanol (volume ratio 2: 1) as eluent gave no stains at Rf 0.31, which corresponds to the starting material, but a main stain at Rf 0.63.

1 ml of water was added to the reaction mixture and after standing for 5 minutes the mixture was treated with “Amberlite IR-120” (H + form) in order to remove the sodium ion therefrom. After distillation under reduced pressure to remove the solvent used and subsequent digesting with ethyl ether, a hygroscopic amorphous solid mass (264 mg) was obtained. Yield: 75.5%. Melting point:

84 to 85 ° C (with decomposition), [a] 2 £ -8.0 ° (c = 0.5, H20).

Elemental analysis: calculated for C8H14

C 33.87,

found: C 34.17,

N30 (, C1:

H 4.97, H 5.10,

N 14.81, N 14.44,

CI 12.50% Cl 12.80% '.

(c) 1- (2-Chloroethyl) -i-nitroso-3- (2,3,4-tri-O-acetyl-ß-D-xylo-pyranosyO-urea

1 - (2-Chloroethyl) -1-nitroso-3- (ß-D-xylopyranosyl) urea (99 mg) was acetylated with acetic anhydride and pyridine in the same manner as described in Example 1 (b). Crystallization from a mixture of chloroform and n-propanol gave the desired compound (42 mg). Yield: 29.4% '. Melting point: 124.5 to 125 ° C (with decomposition).

[a] ß -19.0 ° (c

0.9, chloroform).

Example 9

(a) 1- (1,3 / 2N-Dihydroxycyelohexyl) -3-methyl-urea 2a-amino-β, 3β-cyclohexanediol (378 mg) was added to water

(10 ml) was dissolved and methyl isocyanate (0.35 ml, 2 molar equivalents) was added to the solution while stirring and cooling with ice. When the addition is complete, stirring is continued for 1.5 hours. The reaction mixture is then concentrated under changed pressure and the residue is recrystallized from ethanol, the desired compound being obtained in the form of needles (368 mg) with a melting point of 170 to 173 ° C. Concentrating the mother liquor under reduced pressure gave another portion of the desired compound (96 mg). Melting point: 169 to 172 ° C. The total yield was 464 mg (86%).

(b) / - (1,3 / 2N-Dihydroxycyclohexyl) -3-methyl-3-nitrosourea

1 - (1,3 / 2N-Dihydroxycyclohexyl) -3-methylurea (308 mg, 1.6 millimoles) and sodium nitrite (190 mg, 2.8 millimoles) were dissolved in 5 ml of water, followed by 1 ml of glacial acetic acid with stirring and ice cooling was added to the solution.

After stirring further for 1 hour while cooling with ice, the reaction mixture was left overnight in a refrigerator.

Pale yellow crystals which had separated from the reaction mixture were collected by filtration and gave 65 mg of the desired compound. Melting point: 65 to 67 ° C (with decomposition). The filtrate was treated with “Amberlite IR-120” (H + form) to remove the sodium cations and then distilled under reduced pressure to remove the solvent used, leaving a crystalline residue. The residue was digested with ethanol to give another portion of the desired compound (203 mg) in the form of pale yellow crystals. Melting point: 64 to 65 ° C (with decomposition). Overall yield: 268 mg (75.4%).

IR spectrum (cm "1): 1715 (amide I), 1545 (amide II), 1475 (N-NO).

Elemental analysis:

calculated for C8H15N304: C44.23, H6.96, N 19.34% found: C 44.61, H 6.96, NI 8.77%.

(c) I- (1,3-Di-O-acetyl-1,3 / 2N-dihydroxycyclohexyl) -3-methyl-3-nitrosourea

1 - (1,3 / 2N-Dihydroxycyclohexyl) -3-methyl-3-nitrosourea (105 mg) was dissolved in a mixture of acetic anhydride (1 ml) and pyridine (1 ml) and the resulting mixture overnight at room temperature ditched. After treating the reaction mixture as in Example 1 (b)

17th

619 967

the crystalline residue obtained was digested with ethyl ether to obtain the desired compound (123 mg). Yield: 84.5%. Melting point: 119 to 120 ° C (with decomposition).

Example 10

(a) 1- (2-C'hloriithyl) -3- (1,3 / 2N-dihydroxycyciohexyl) urea

2 «-amino-Iß, 3ß-cycIohexandioI (557 mg) was in 2-

Suspended methoxyethanol (30 ml) containing methyl cellosolve, and the resulting mixture was mixed with 2-chloroethyl isocyanate (1 ml, 2.7 molar equivalents) in small portions with stirring, and ice cooling. Stirring was continued for an additional 1.5 hours to obtain a clear solution, which was then concentrated under reduced pressure to give a crystalline residue. Recrystallization from a mixture of methanol and n-i propanol gave 670 mg of the desired compound. Yield: 67%. Melting point 125 to 126.5 ° C (with decomposition).

(b) 1 - (2-ChloroethyI) -3- (1,3 / 2N-dihydroxycyclohexyl) -l-nitro-> urea

1 - (2-chloroethyl) -3- (1,3 / 2N-dihydroxycycIohexyl) urea (426 mg, 1.8 millimoles) was dissolved in 99% formic acid (6 ml) and then sodium nitrite (204 mg, 3.0 millimoles) was added to the solution while stirring and cooling with ice, and the resulting mixture was then stirred with ice cooling for a further 2 hours.

The reaction product was a clear yellow solution, whose thin layer chromatography on silica gel using chloroform-ethanol (6: 1 by volume) as eluent gave no stain at Rf 0.51, which corresponds to the starting compound, but a main stain at Rf 0.71.

The reaction mixture was mixed with 2 ml of water and the mixture obtained after standing for 5 minutes with «Amberlite! IR-120 »(H + form) treated to remove the sodium cations and then distilled under reduced pressure to remove the solvent used. Recrystallization of the residue obtained from a mixture of ethanol and ethyl ether gave the desired compound (281 mg) as pale yellow crystals. Yield: 58.7%. Melting point: 125 to 125.5 ° C (with decomposition). IR spectrum (cm '): 1700 (amide I), 1555 (amide II), 1500 (N-NO).

I.

Elemental analysis:

calculated for C9Hlf) N304Cl:

C 40.68, H 6.07, N 15.82, Cl 13.34% found: C40.51, H5.87, N 15.70, Cl 13.13%.

S '

(c) / - (2-chloroethyl) -3- (1,3-di-O-acetyl-l, 3 / 2N-dihydroxycy-clohexyl) -! - nitrosourea l- (2-chloroethyl) -3- (l , 3 / 2N-dihydroxycyclohexyl) -l-nitrosourea (1 16 mg) was dissolved in a mixture of acetic anhydride (1 ml) and pyridine (1 ml) and the resulting mixture was left overnight at room temperature. The reaction mixture was then distilled under reduced pressure to remove the solvent used, whereby a crystalline residue was obtained, which was digested with ethanol to give the desired compound (132 mg). Yield: 86.4%. Melting point: 151 to 152 ° C (with decomposition).

Example II

1- (β-D-Mannopyranosyl) -3-methyl-3-nitrosourea "

1- (β-D-Mannopyranosyl) -3-methyl-urea (200 mg) was dissolved in a mixture of 1.5 ml of glacial acetic acid and 3.5 ml of water and the solution obtained was mixed with 99 mg of sodium nitrite. The mixture thus obtained was shaken at room temperature for 3 hours to achieve nitrosation. The reaction mixture was treated with “Amberlite IR-120” (H + form) to remove the sodium cation. The reaction mixture thus treated was then concentrated under reduced pressure at a temperature below 30 ° C to remove the solvent, whereby a crystalline product was precipitated. This crystalline product was filtered off and washed with n-propanol to give 180 mg of the desired compound. Yield: 78%.

Melting point: 103 ° C (with decomposition), [tx] (c = 0.99, H20).

Elemental analysis:

calculated for CKH15N, 07: C 36.23, H 5.70, found: C 35.92, H 5.70,

12.6 °

N 15.84% N 15.67%.

Example 12

Methyl-2, fi-dideoxy-2- (N '-methyl-N' -nitrosoureido) -ß-D-glu-copyranoside

Methyl 2,6-dideoxy-2- (N'-methyl-ureido) -ß-D-glucopyra-noside (124 mg) of the formula

OCH-

NHCNHCH.

was dissolved in a mixture of 0.5 ml of glacial acetic acid and 2.5 ml of water, and the resulting solution was mixed with 55 mg of sodium nitrite at room temperature and with shaking to achieve nitrosation. The reaction mixture thus obtained was then left in a refrigerator overnight, whereby a crystalline product was separated. This crystalline product was collected by filtration and washed with ethanol to obtain 80 mg of methyl 2,6-dideoxy-2- (N'-methyl-N'-nitroso-ureido) -ß-D-glucopyranoside as the desired product. The mother liquor was then treated with “Amberlite IR-120” (H + form) to remove the sodium ion and mother liquor treated in this way was concentrated by distilling off the solvent under reduced pressure. Crystallization of the residue from ethanol gave 45 mg of the desired compound second

28

Portion. Yield: 125 mg (90%). [cx] "previous year H20).

14 ° (c = U, 52,

Elemental analysis:

calculated for CgH17N30 (1: C41.06, H 6.51, found: C 41.03, H 6.42,

N 16.00 '

Example 13

Methyl 2-deoxy-6-0-mesyl-2- (N '-methyl-N' -nitroso-ureido) -ß-D-glucopyranoside

Methyl 2-deoxy-6-0-mesyl-2- (N'-methylureido) -ß-D-glu-copyranoside (400 mg) of the formula

619 967

18th

was dissolved in 50 ml of water and the aqueous solution obtained was adjusted to pH 3 by adding 2 ml of acetic acid. The acidified aqueous solution was mixed with 120 mg of sodium nitrite and the mixture was shaken at room temperature overnight to achieve the nitrosation. The reaction mixture was treated with "Amberlite IR-120" (H + form) to remove the sodium ion. The reaction mixture was then concentrated under reduced pressure at a temperature below 25 ° C. The residue was crystallized from ethanol and gave 350 mg of the desired compound.

oa

Melting point: 131 to 132 ° C (with decomposition), [a] "p - 14 ° (c = 0.5, dimethyl sulfoxide).

Elemental analysis:

calculated for C1 () H19N3S09:

C 33.61, H 5.35, found: C 33.87, H 5.36,

was dissolved in 25 ml of 99% formic acid and 262 mg (3.8 millimoles) of sodium nitrite were added in small portions to the solution thus obtained. The mixture thus obtained was stirred under ice-cooling for 2 hours to complete the nitrosation. The reaction mixture was mixed with 2 ml of 1 water, left to stand for 5 minutes and then treated with “Amberlite IR-120” to remove the sodium cations. The mixture thus treated was evaporated to dryness by evaporating the solvent under reduced pressure to obtain a crystalline residue. This residue was digested with ethanol to give 636 mg of the desired compound as pale yellow crystals. Yield: 77.9%. Melting point: 140 to 142 ° C (with decomposition).

IR spectrum: 1705 (Amidi), 1555 (AmidII) and 1500cm "1 (N-NO).

N 11.75, N 11.44,

S 8.97% S 8.73%.

Elemental analysis:

calculated for C9H16N307C1:

C 34.46, H 5.14, N 13.40,

found: C 34.66, H 5.15, NI 1.50,

Cl 11.30% Cl 11.50%.

Example 14

(a) 1- (2-Chloroethyl) -3- (1N, 3.5 / 2.4, fi-pentahydroxycycìohexyl) urea

A solution of scyllo-inosamine (551 mg) in water (20 ml) was cooled in an ice bath. To this cold solution, 0.6 ml (2.3 molar equivalents) of 2-chloroethyl isocyanate were added in small portions while cooling with ice and stirring. The mixture thus obtained was further stirred for 2 hours to complete the reaction. The precipitated crystalline product was separated from the reaction mixture by filtration and washed with ethanol and gave 432 mg of the desired product as crystals of melting point 195 to 197 ° C. (with decomposition). The filtrate from which the crystalline product was removed was then concentrated by evaporation of the solvent under reduced pressure, and the residue thus obtained was digested with ethanol. 357 mg of the desired compound were obtained as a second portion. Overall yield: 90% (789 mg). «

IR spectrum (in KBr): 1625 (C = 0) and 1580 cm "1 (NH,

Amide II).

Elemental analysis: Ml calculated for C9H17N206C1:

C 37.97, H 6.02, N9.84, Cl 12.45% found: C 37.96, H 5.78, N9.76, Cl 12.71%.

(b) 1 - (2-chloroethyl) -1-nitroso-3- (1 N, 3,5 / 2,4,6-pentahydroxy-cyclohexyl) urea l- (2-chloroethyl) -3- (lN, 3,5 / 2,4,6-pentahydroxycyclo-hexyl) urea (741 mg, 2.6 millimoles) of the formula

(c) 1- (2-Chloroethyl) -l-nitroso-3- (2,3,5,6-penta-O-acetyl-1N, 3,5 / 2,4,6-pentahydroxycyclohexyl) urea

1 - (2-Chloroethyl) -1-nitroso-3- (1N, 3,5 / 2,4,6-pentahydroxycyclohexyl) urea (116 mg) was described in the same manner as in Example 1 (b) , acetylated by reaction with acetic anhydride and pyridine. The acetylation product was crystallized from ethanol to give 171 mg of the desired compound as pale yellow crystals. Yield: 88.3%. Melting point: 205.5 to 209 ° C (with decomposition).

Elemental analysis:

calculated for C, 9H26Oj2N3CI:

C 43.56, H 5.00, found: C 43.58, H 4.97,

N 8.02, N 7.79,

Cl 6.77% Cl 6.75%.

Example 15 (a) 1- (β-D-Maltosyl) -3-methylurea

To a solution of 2.5 g 2,3,6,2 ', 3', 4 ', 6'-hepta-0-acetyl-ß-D-maltosylamine (prepared as ß-D-maltosylamine according to the method by A. Bertho in "Ann.", 562.229 (1949)) in 40 ml of dioxane, 0.7 ml of methyl isocyanate was slowly added with ice cooling and stirring. The mixture was stirred for an additional 3 hours and the reaction mixture was then concentrated under reduced pressure. Crystallization of the residue from isopropanol gave 2.33 g of l- (β-D-maltosyl) -3-methylurea-hepta-0-acetate. This compound (2.0 g) was dissolved in 50 ml of ammonia-saturated methanol at 0 ° C and the mixture was left at room temperature for 2 days to complete the reaction. The reaction mixture was reduced

19th

619 967

concentrated pressure and the crystallization of the residue from methanol-ethanol gave the desired compound. Yield: 665 mg (57.8%). Melting point: 90 to 92 ° C.

M d'5 + 70.0 ° (c = 0.5, H20).

IR spectrum: 1660 and 1570cm ^ '(Ureido).

(b) 1- (β-D-Maltosyl) -3-methyl-3-nitrosourea

1 - (β-D-Maltosyl) -3-methylurea (300 mg) was dissolved in a mixture of 2 ml of water and 0.5 ml of glacial acetic acid and 100 mg of sodium nitrate were slowly added to the resulting solution with ice cooling and stirring . The solution was stirred overnight with water cooling to complete the nitrosation. The reaction mixture was then treated with a cation exchange resin, “Amberlite IR-120”, to remove the sodium ions and then concentrated under reduced pressure. Crystallization of the residue from ethanol-ethyl ether gave the desired compound. Yield: 293 mg (91.0%). Melting point:

128 ° C (decomposition under foaming), [a] ^ + 90 ° (c = 0.5, "water).

IR spectrum: 1730, 1550 (ureido) and 1490 cm-1 (nitrosamin).

Example 16

(a) / - (2-chloroethyl) -3- (ß-D-maltosyl) urea

The procedure of Example 15 (a) was repeated using 2,3,6,2 ', 3', 4 ', 6'-hepta-0-acetyl-β-D-matosylamine and 2-chloroethyl isocyanate. The desired compound was obtained in a yield of 46.1%. Melting point: 108 ° C (with decomposition).

(b) 1 - (2-Ch! ethyl) -3- (ß-D-maltosyl) -l-nitrosourea

150 mg of sodium nitrite were slowly added to a solution of 500 mg of 1 - (2-chloroethyl) -3- (β-D-mal-tosyl) urea in 3 ml of aqueous 99% formic acid with ice cooling and stirring. The mixture was stirred for 2 hours and the reaction mixture was then treated with "Amberlite IR-120" (H + form) to remove the sodium cation.

After removal of the cation exchange resin from the reaction mixture by filtration, the filtrate was concentrated under reduced pressure. Crystallization of the residue from ethanol-isopropanol gave the desired nitroso compound. Yield: 227 mg (52.0%). Melting point: 96 ° C

(Decomposition under foaming), [a] ^ + 66 ° (c = 0.5, water). , s

IR spectrum: 1730.1540 (ureido) and 1505 cm "1 (nitrosamin).

Example 17 mi

(a) / - () 8-D-Lttctosyl) -3-methylurea The procedure of Example 15 (a) was repeated using

Use of 2,3,6,2 ', 3', 4 ', 6'-hepta-0-acetyl-ß-D-lactosylamine and methyl isocyanate. The desired connection was obtained.

(b) l- (β-D-lactosyl) -3-methyl-3-nitrosourea

150 mg of sodium nitrite were slowly added to a solution of 500 mg of l- (β-D-lactosyl) -3-methyl-urea in a mixture of 5 ml of water and 0.5 ml of glacial acetic acid while cooling with ice and stirring. The mixture was stirred overnight with cooling with water and the reaction mixture was treated with “Amberlite IR-120” (H + form) to remove the sodium cations. After filtering the reaction mixture to remove the resin, the filtrate was concentrated under reduced pressure. Crystallization of the residue from methanol-ethanol gave the desired compound as pure crystals. Yield: 440 mg (82.0%). Melting point:

175 ° C (decomposition with foams), [a] ^ + 4.0 ° (c = 0.5, water).

IR spectrum: 1725, 1540 (ureido) and 1485 cm'1 (nitrosamin).

Example 18

(a) 1- (2-Chloroethyl) -3- (β-D-lactosyl) urea

The procedure of Example 15 (a) was repeated using 2,3,6,2 ', 3', 4 ', 6'-hepta-0-acetyI-β-D-lacto -sylamine and 2-chloroethyl isocyanate. The desired connection was obtained. Melting point: 119 to 120 ° C.

(b) l- (2-Chloroethyl) -3- (ß-D-lactosyI) -l-nitrosourea

150 mg of sodium nitrite were slowly added to a solution of 500 mg of 1- (2-chloroethyl) -3- (β-D-lac-tosyI) urea in 300 ml of 99% aqueous formic acid with ice cooling and stirring. The mixture was then processed in the same manner as described in Example 17 to obtain the desired compound. Yield: 290 mg (54.5%). Melting point: 129 ° C (decomposition under foaming).

[«]

22.5 D.

+ 4.0 ° (c = 0.5, water).

IR spectrum: 1735, 1540 (ureido) and 1500 cm "1 (nitrosa-min). The corresponding D-galactosoaminol can also be prepared in an analogous manner.

Example 1V

(a) N-Carbamoyl-N '- (2-chloroethy!) - D-glucosaminol

A solution of 500 mg of D-glucosaminol in 8 ml of water was mixed with 1.5 ml (5 molar ratios) of 2-chloroethyl isocyanate with stirring and cooling with water. The mixture was stirred under water cooling for 1 day. The reaction mixture was then filtered under vacuum to remove the solids including the by-products. The filtrate was mixed with 12 ml of a cation exchange resin, “Amberlite IR-120” (H + form) and then separated from the resin by filtration. The filtrate was concentrated under reduced pressure and the residue was taken up in methanol. The addition of ethyl ether to the methanolic solution caused the excretion of a gel-like solid. This colorless solid was dissolved in a solution of sodium methoxide in methanol, then left to stand under ice cooling for 1 day and then mixed with “Amberlite IR-120” (H + form). After the mixture reached pH 4, it was filtered to remove the resin. The filtrate was concentrated under reduced pressure to give a residue, which was then crystallized from ethanol. The desired compound was obtained in the form of crystals in a yield of 360.6 mg (45.1%)

receive. Melting point: 124 to 125 ° C. [a] ^ (c = 1.0, methanol).

IR spectrum: 1620 (amide I) and 1580 cm "1 (amide II).

(b) N-Carbamoyl-N '- (2-chloroethyl) -N-nitroso-D-glucosaminoi A solution of 50 mg of N-carbamoyl-N' - (2-chloroethyl) -D-

Glucosaminol in 1 ml glacial acetic acid was mixed with 27 mg (2.25 molar ratios) of sodium nitrite with ice cooling and stirring and the mixture was further stirred for 4 hours with ice cooling to complete the nitrosation. The reaction mixture was filtered under vacuum to remove the solids including the by-products. The filtrate was mixed with 2 ml of "Amberlite IR-120" (H + form). After the mixture reached pH 1, it was filtered to remove the resin. The filtrate was freeze-dried to give the desired nitroso compound. Yield: 31 mg (56.3%). Melting point: 73 to 74 ° C.

619 967

20th

[a] 2p + 22, S "C (c = 0.5, methanol).

IR spectrum: 1710 (Amidi), 1540 (Amid II) and 1490 cm 1 (N-nitroso group).

IR spectrum: 1700 (ester), 1630 (amide I) and 1590 cm 1 (amide II).

Elemental analysis:

calculated for C \, HIS07N3C1:

C 34.24, H 5.74, found: C 33.22, H 5.45,

N 13.31, N 12.52,

Cl 11.23% CI 10.79%.

Elemental analysis:

calculated forCKHi503N, CI2:

C 35 ^ 31, H 5.56, found: C 35.54, H 5.52,

N 15.44, N 15.36,

Cl 26.05% CI 26.07%.

Example 20

(a) (N-Carbamoyl-N'-methyl) -O- (O-carbamoyl-N-methyl) ethanolamine

To a solution of 10 ml of ethanolamine in 40 ml of methanol was gradually added 24 ml (2.5 molar ratios) of methyl isocyanate in an ice bath. The mixture was shaken for 3 hours to separate out a crystalline product. The whole mixture was left in an ice box overnight and then filtered, whereby the desired compound was obtained as white crystals. Yield: 11.11 g (38.3%), melting point: 133 to 134 ° C.

[uJ22-4 ° (c = 1.0, H20).

IR spectrum: 1700 (ester, 1630 (C = 0-amide I) and 1590 cm "1 (amide II).

Elemental analysis:

calculated for C (1H,., N303: C 41.14, H 7.48, N 23.99%

found:

C 41.25, H 7.32, N 24.16%.

(b) (N-Carbamoyl-N'-methyl-N'-nitroso) - (O-carbamoyl-N-methyl) ethanolamine

A solution of 1.73 g (2.2 molar ratios) of sodium nitrite slowly became a solution of 2 g of (N-carbamoyl-N'-methyl) -O- (O-carbamoyl-N-methyl) ethanolamine in 2 ml of glacial acetic acid added in 45 ml of water in an ice bath. The mixture was stirred for 1 hour to complete the nitrosation. The reaction mixture was then filtered to separate the separated white colored crystals. The filtrate was mixed with 50 ml of a cation exchange resin, “Amberlite IR-120” (H + form). After the mixture reached pH 2, it was separated from the resin by filtration. The filtrate was evaporated to dryness by evaporating the solvent at a temperature below 30 ° C under reduced pressure. The residue was crystallized from acetone and gave yellow-colored needles. Recrystallization of the crystals from methanol gave the desired compound. Yield: 1.694 g (72.7%). Melting point: 97 ° C (decomposition under foaming), [ot] ^ - 0.4 ° (c = 1.0, methanol).

IR spectrum: 1730 (ester and amide I), 1530 (amide II) and 1480 cm "" 1 (N-nitroso group).

Elemental analysis: calculated for C6H12N404: found:

C 35.29, H 5.93, C 35.45, H 5.85,

Example 21

(a) (N-Carbamoyl-N-2-chloroethyl) - (0-carbamoyl-N-2-chloroethyl) ethanolamine

The procedure of Example 20 (a) was repeated using 2 ml of ethanolamine and 4 ml of 2-chloroethyl isocyanate. The desired compound was obtained in a yield of 1.999 g (22.4%). Melting point: 124 to 126 ° C.

[a] 2q + 64 ° (c = 1.0, acetone).

(b) (N-Carbamoyl-N'-2-chloroethyl-N'-nitroso) - (D-carbamoyl-i ti N '-2-chloroethyl) ethanolamine

To a solution of 200 mg (N-carbamoyl-N'-2-chloro-ethyI) - (0-carbamoyl-N'-2-chloro-ethyI) -ethanolamine from Example 20 (b) in 4 ml of glacial acetic acid was slowly added 200 mg ( 3.9 molar ratios) sodium nitrite added in an ice bath. The mixture was stirred under ice cooling for 2 hours to complete the nitrosation. The reaction mixture was then filtered to remove the insoluble substances and the filtrate was mixed with 14 ml of "Amberlite IR-120" (H + form). After the mixture reached pH 1, it was filtered to remove the resin. The filtrate was concentrated under reduced pressure and the resulting oily residue was crystallized from a mixture of chloroform and isopropanol, whereby the desired compound was obtained as yellow-colored crystals. Yield: 149.7 mg

(67.6%). Melting point: 64 to 65 ° C. [a] 2p + 4 ° (c = 0.5, methanol).

IR spectrum: 1700 (ester and amide I), 1530 (amide II) and 1490 cm "1 (N-nitroso group).

<"Elemental analysis:

calculated for C8H1404N4C12:

C 31.90, H 4.69, N 18.61,

found: C32.21, H4.69, N 18.32,

N 27.44% N 27.75%.

Cl 23.55% Cl 23.34%.

Example 22

(a) 1- (2,3,4-Tri-0-acetyl-fi-0-tosyl-β-D-g / ucopyranosy /) - S-methylurea

A suspension of 1 - (β-D-glucopyranosyl) -3-methyl-urea (3.23 g, 13.7 millimoles) in pyridine (50 ml) was washed with a solution of tosyl chloride (3.12 g, 16.4 Millimol) in pyridine (6 ml) and the mixture was stirred for 18.5 hours at 5 ° C. with ice cooling and then for a further 43 hours at room temperature, whereupon a further portion of the solution of tosyl chloride (1.40 g, 7.3 Millimoles) in 3 ml of pyridine was added. The mixture was stirred for an additional 23 hours. Thin layer chromatography of the reaction mixture on silica gel using chloroform-methanol (2: 1 by volume) as eluent gave no stain at Rf 0.43, which corresponds to the starting compound, but a major stain at Rf 0.75.

The reaction mixture was poured into 800 ml of ice water and left in the cold overnight. A syrupy phase had separated out and the supernatant aqueous layer was removed by decantation and evaporated to dryness under reduced pressure. The residue was then subjected to acetylation by reacting with acetic anhydride in pyridine to give 5.47 g of a syrup. This syrup was subjected to column chromatography with 100 g of silica gel and using benzene-isopropanol (volume ratio 10: 1) as the eluent. Those fractions of the eluate which gave a single spot at Rf 0.39 were pooled and evaporated to dryness by evaporation of the solvent under reduced pressure. Crystallization of the residue from n-propanol gave the desired compound in a yield of 2.20 g

(31.1%). Melting point: 118.5 to 120 ° C. [a] 2 pounds + 20.1 ° (c = 2.45, chloroform).

21st

619 967

(b) 1 - (2,3,4-Tri- () -acetyl-6-deoxy-6-iodo-ß-D-glucopyra-nosyI) -3-methylurea

A solution of 2.38 g (4.6 millimoles) of 1 - (2,3,4-tri-O-aceto-tyl-6-0-tosyl-ß-Dg! UcopyranosyI) -3-methylurea in 15 ml of dioxane was mixed with 6.64 g (44.3 millimoles) of sodium azide - and the mixture was stirred at 100 ° C. for 3 hours and then cooled to room temperature. The reaction mixture was added to a mixture of 100 ml of chloroform and 20 ml of water with stirring. The chloroform layer was washed with saturated aqueous solution of sodium chloride, then with aqueous 20% sodium thiosulfate solution and then with water and then dried over anhydrous sodium sulfate. The chloroform layer treated in this way was then evaporated to dryness under reduced pressure, giving 3.90 g of an amorphous solid as an i-residue. This solid was subjected to column chromatography with 100 g of silica gel using benzene-n-propanol (volume ratio 10: 1) as the eluent. Those fractions of the eluate which gave a single spot at Rf 0.46 were pooled and evaporated to dryness by evaporation of the solvent under reduced pressure. The residue was taken up in methanol and then twice the volume of water was added, the desired compound being precipitated as crystals. Yield: 1.47 g (67.6%). Melting->

point: 144 to 147 ° C. [a] 2 ^ + 2.4 ° (c = 1.4, chloroform).

(c) 1- (2,3,4-Tri-0-acetyl-6-deoxy-β-D-glucopyranosyl) -3-methylurea

1 - (2,3,4-Tri-0-acetyl-6-deoxy-6-iodo-β-D-glucopyrano-syl) -3-methylurea (661 mg) was dissolved in 20 ml of ethyl acetate and subjected to catalytic hydrogenation during 2 hours in the presence of 7 ml of "Amberlite IR-45" (OH form) and in the presence of a Raney nickel catalyst at room temperature and at an initial hydrogen pressure of 3.4 kg / cm2 to remove the azido group in to transfer the amino group.

Thin layer chromatography of the reaction mixture on silica gel using benzene-isopropanol (volume ratio 6: 1) as the eluent gave no stain at Rf 0.39, which corresponds to the starting compound, but a single stain at Rf 0.32.

After removal of the catalyst and the ion exchange resin by filtration, the filtrate was evaporated to dryness under reduced pressure. The recrystallization of the crystalline residue obtained from ethanol gave the desired compound. Yield: 341 mg (70.3%). Melting point: 180 to 182.5 ° C. [a] + 22.4 ° (c form).

0.56, chloro-

(d) I- (6-deoxy-β-D-glucopyranosyl) -3-methylurea

A solution of 1.42 g of l- (2,3,4-tri-0-acetyl-6-deoxy-ß-ss D-glucopyranosyl) -3-methylurea in 6 ml of methanol was mixed with 1 ml of 1N sodium methoxide in methanol and the mixture was left to stand at room temperature for 22 hours.

The reaction mixture was then neutralized by adding a "cation exchange resin," Amberlite IR-120 "(H + form), filtered off from the resin and then concentrated to a syrupy residue by evaporation of the solvent under reduced pressure. Crystallization of the residue from ethanol gave the desired compound. Yield: 0.66 g (73.1%). Melting point: 194 to

196 ° C. [A] 2 ^ -23.9 ° (c = 1.0, water).

(e) / - (fi-deoxy-ß-D-glucopyranosyl) -3-methyl-3-nitrosourea l- (6-deoxy-ß-D-glucopyranosyl) -3-methylurea (617 mg, 2.8 Millimoles) and sodium nitrite (242 mg, 3.5 millimoles) were dissolved in 4 ml of water and the resulting solution was mixed with 0.5 ml of glacial acetic acid while cooling with ice and stirring. The mixture was stirred under ice cooling for 1 hour and then left in a refrigerator overnight to complete the nitrosation.

i The reaction mixture was treated with “Amberlite IR-120” (H + form) to remove the sodium cations and was then evaporated to dryness by evaporating the solvent under reduced pressure. The digestation of the crystalline residue obtained with ethanol gave the desired nitroso compound. Yield: 516 mg (71.3%). Melting point: 76 to 77 ° C (with decomposition).

[cx] 2 ^ -24.0 ° (c = 1.1, water).

IR spectrum (in KBr): 1745 (C = 0, amide I), 1535 (CNH, i amide II) and 1470 cm-1 (N = 0).

Elemental analysis:

calculated for CxH15N, Oh • '/: H20:

C 37.21, H 6.25, N 16.27%. found: C 36.94, H 6.11, N 16.66%.

Example 23

(a) / - (2-chloroethyl) -3- (fi-deoxy-ß-D-glucopyranosy /) - urea

A solution of 234 mg (0.91 millimoles) l- (6-deoxy-ß-D-

glucopyranosyl) -3-methyl-3-nitrosourea in 2 ml water was treated with a solution of 401 mg (3.5 millimoles) 2-chloroethylamine hydrochloride and 0.5 ml (3.3 millimoles) triethylamine in 3 ml aqueous 50% mixed ethanol. The mixture was stirred at room temperature for 2 hours and the reaction mixture was then passed over a column (10 × 160 mm) “Aberlite IRA-400” (OH ~ form), which was then washed with water. The outflowing solution was evaporated to dryness under reduced pressure and the crystalline residue thus obtained was subjected to chromatography in a column (8 × 350 mm) silica gel (8 g) using chloroform-methanol (5: 2 by volume) as the eluent. Those fractions of the eluate which gave a single spot at Rf 0.50 were combined and the evaporation of the solvent evaporated to dryness under reduced pressure. Recrystallization of the residue from acetone gave the desired compound. Yield: 98 mg (37.7%). Melting point: 130 to 132 ° C.

[«] -25.5 ° (c = 0.5, water).

(b)] - (2-chloroethyl) -3- (fi-deoxy-ß-D-glucopyranosyl) -! - nitrosourea

A solution of 193 mg (0.67 millimoles) of 1- (2-chloroethyl) -3- (6-deoxy-ß-D-glucopyranosyl) urea in 1.5 ml of aqueous 99% formic acid was treated with 99 mg of sodium nitrite Ice cooling and stirring mixed. The mixture was stirred with ice cooling for 3 hours to complete the nitrosation. The reaction mixture was then mixed with 0.5 ml of water and then left to stand for 15 minutes, after which it was treated with “Amberlite IR-120” (H + form) to remove the sodium cations. The reaction mixture was then concentrated to a syrupy residue by evaporating the solvent under reduced pressure. The syrup was subjected to chromatography on a column (8 X 300 mm) silica gel (7 g) using chloroform-methanol (4: 1 by volume). Those fractions of the eluate which gave a single spot at Rf 0.58 were pooled and under

619 967

22

evaporated to dryness to give 145 mg (72.3%) of the desired nitroso compound as a glassy but hygroscopic solid of pale yellow color, [a] ^ -5.6 ° (c = 0.8, water).

The nitroso compound obtained (75 mg) was acetylated by reaction with acetic anhydride in pyridine. The reaction mixture from the acetylation was evaporated to dryness under reduced pressure, whereby a crystalline residue was obtained. The digesting of this residue with isopropanol gave 57 mg of l- (2-chloroethyl) -3- (2,3,4-tri-0-acetyl-β-D-glucopyranosyl) -l-nitrosourea as crystals. Yield: 53.4%. Melting point: 143 to 144 ° C.

Example 24

Tetra-N- (N -methyl-N -nitroso) -carbamoyl-ribostamycin

Tetra-N- (N'-methyl) carbamoyl ribostamycin (1.0 g)

was dissolved in 30 ml of ice-cold water and the solution obtained was mixed with 0.6 g of sodium nitrite. To the mixture thus obtained, which was in the form of a solution, 2.0 ml of glacial acetic acid was added dropwise with ice cooling and stirring. The mixture was stirred continuously for 2 hours and then left overnight at 0 ° C to complete the nitrosation. The reaction solution was evaporated in vacuo below 25 ° C. The residue was taken up in a small volume of methanol and the ethanolic solution obtained was again evaporated to dryness. The residue thus obtained was taken up in about 20 ml of anhydrous methanol and the small amount of insoluble matter was filtered off. The filtrate was evaporated to a volume of about a quarter of the original volume by evaporation of the solvent under reduced pressure.

The concentrated solution was cooled to -20 ° C, whereby a pale yellow amorphous product was excreted. The excreted product was filtered off and the mother liquor was mixed with twice the amount of ethyl ether and then cooled in order to separate another portion of the amorphous product. The precipitated product was collected by filtration and gave 0.68 g of the desired compound as an amorphous and hygroscopic solid of a pale yellow color. Melting point: 88 ° C (decomposition under foaming).

[e] + ^ 5 ° (c = 1.0, methanol).

Elemental analysis:

calculated for C2JH42N12OiX: C 37.59, H 5.30, N 21.05%

"'Found: C 36.61, H 5.40, N 19.87%.

Example 25

Tetra-N-fN- (2-chloroethyl) -N -nitrosoj-carbamoyl-ribosta-mycin

Tetra-N- [N '- (2-chloroethyl)] - carbamoyl-ribostamycin (1.9 g) was dissolved in 30 ml of cold water and the resulting solution was mixed with the same volume (30 ml) of glacial acetic acid. To this mixture, 1.1 g of sodium nitrite was gradually added while cooling with ice and stirring. The mixture thus obtained was stirred under ice-cooling for 2 hours to complete the nitrosation. The reaction mixture was treated with “Amberlite IR-120” (H + form) to remove the sodium ions. The reaction mixture was then filtered and the filtrate evaporated under reduced pressure at a temperature below 25 ° C. The residue was dissolved in ethanol and the solution was again concentrated in the same manner as above. The residue thus obtained was taken up in a small amount of ethanol, and a small volume of ethyl ether was added to the solution obtained, so that the desired compound was precipitated. The precipitate was collected by filtration, washed with ethyl ether and dried in an excicator. The desired compound was obtained in a yield of 0.9 g. Melting

"point: 105 ° C (decomposition under foaming), [a] ^ + 30.7 ° (c = 1.0, methanol).

C.

Claims (34)

619 967 PATENT CLAIMS 1. Process for the preparation of new nitrosourea derivatives of the formula 0, N0 A ■ NHC- ■ N A- 0 II ■ NHCN 'NO' R n 'R 1 /, ï 1 (Ia) (I) in which A1 represents the β-D-glucopyranosyl group and R1 represents an alkyl group having 2 to 4 carbon atoms or a halogen-substituted alkyl group having 1 to 4 carbon atoms. , 2 (If) a6— nhcnr _ r3 ocnhr4 'R in which Afi represents a glycosyl group derived from a disaccharide and R2 represents an alkyl group with 1 to 4 carbon atoms or a halogen-substituted alkyl group with 1 to 4 carbon atoms. 2 o 'R a —nhcn: (Ih) in which A5 the pentahydroxycyclohexyl group and R2 a ^ sr2 Represents alkyl group having 1 to 4 carbon atoms or a halogen-substituted alkyl group having 1 to 4 carbon atoms. 2nd ch- (CHOH) n has ch2oh, in which n is zero or an integer from 1 to 3 and R2 is a halogen-substituted alkyl group having 1 to 4 carbon atoms. 2. The method according to claim 1, characterized in that the nitrosating agent is an alkali metal nitrite. 2 ii A NHCN 0! NHCNHR (") n in which A, R and n have the same meaning as above, reacting a with a nitrosating agent. 3rd 619 967 3. The method according to claim 1, characterized in that the nitrosating agent nitrogen trioxide or Distick-? Substance tetroxide is. 4th A NHCN (Ii) in which A '' denotes the 6-deoxyglucopyranosyl group and R2 denotes an alkyl group with 1 to 4 carbon atoms or a halogen-substituted alkyl group with 1 to 4 carbon atoms. 4. The method according to claim 1, characterized in that the nitrosation is carried out at a temperature of - 10 ° C to 30 ° C. 5 619 967 chloräthyI) -N'-nitrosol] - (0-carbamoyI-N-2-chloroethyl) -ätha-nolamine. 5. The method according to claim 1, characterized in that the nitrosation is carried out at a pH of 1 to 3. 6. The method according to claim 1, characterized in that the nitrosation is carried out in an inert solvent. ( 7 ... i / N0 (lg) SR CH20Ms OCH, in which Ms represents the mesyl group. 7. The method according to claim 6, characterized in that the inert solvent is water, a lower fatty acid or a mixture thereof. 8. The method according to claim 7, characterized in that the lower fatty acid is formic acid, acetic acid or <propionic acid. 9. The method according to any one of claims 1 to 8 for the preparation of compounds of the formula .NO • R (Ib) in which A2 represents the mannopyranosyl group or the galactopyranosyl group and R2 represents an alkyl group with 1 to 4 carbon atoms or a halogen-substituted alkyl group with 1 to 4 carbon atoms. 10th -NHCN (Ij) in which A1 "is a tetravalent radical of ribostamycin of the formula HÖ \ OH and R2 represents an alkyl group having 1 to 4 carbon atoms or a halogen-substituted alkyl group having 1 to 4 carbon atoms. 10. The method according to any one of claims 1 to 8, for the production of nitrosourea derivatives of the formula in which A is a glycosyl group, a monovalent radical of methylglucoside, from which the 2-hydroxy group has been removed, a monovalent radical of an alditol, from which the 2-hydroxyl group an N-substituted carb-amoyloxyalkyl group or a hydroxy-substituted cyclohex-yl group when n is 1 or a tetravalent radical of ribostamycin from which the four amino groups have been removed when n is 4 and R is a lower alkyl group or one represents halogen-substituted lower alkyl group, where R is not a methyl group, if A is a glucosyl group, characterized in that a compound of the formula 0 11. The method according to any one of claims 1 to 8 for the production of nitrosourea derivatives of the formula 0 , N0 «A * • NHCN • R (Ic) in which A3 represents a glycosyl group or a hydroxy-substituted cyclohexyl group and R2 represents an alkyl group with 1 to 4 carbon atoms or a halogen-substituted alkyl group with 1 to 4 carbon atoms. 12. The method according to claim 11, characterized in that the glycosyl group is the arabinosyl, xylosyl, lyxosyl, talosyl, idosyl, gulosyl, altrosyl or allosyl group. 13. The method according to claim 11, characterized in that the hydroxy-substituted cyclohexyl group is a mono-, di-, tri- or tetrahydroxycyclohexyl group. 14. The method according to any one of claims 1 to 8 for the preparation of nitrosourea derivatives of the formula 0 -NO A - NHC - N (id) * CH, in which A4 the mannopyranosyl group, the methyl-2,6-dideoxy-glucopyranoside residue of the formula 15. The method according to any one of claims 1 to 8 for the preparation of nitrosourea derivatives of the formula in which A7 is a monovalent residue of alditol from which the 2-hydroxy group has been removed and which has the formula or the methyl-2-deoxy-6-0-mesyl -ß-D-giucopyranosiderest ,, of the formula CHo0H 16. The method according to any one of claims 1 to 8 for which Ax is an N-substituted carbamoyloxyalkyl group. Preparation of nitrosourea derivatives of the formula ^ the formula 0, no q 17. The method according to claim 16, characterized in that the glycosyl group is the maltosyl, lactosyl, sucrosyl, cellobiosyl, trehalosyl, gentiobiosyl, melibiosyl, turanosyl, sophorosyl or isosucrosyl group. in which R3 denotes an alkyl group with 2 to 5 carbon atoms and R4 denotes an alkyl group with 1 to 4 carbon atoms or a halogen-substituted alkyl group with 1 to 4 carbon atoms, and R2 denotes an alkyl group with 1 to 4 carbon atoms or a halogen-substituted alkyl group with 1 to 4 carbon atoms Represents 4 carbon atoms. 18. The method according to any one of claims 1 to 8 for the preparation of nitrosourea derivatives of the formula OCH, 19. The method according to claim 18, characterized in that the monovalent residue of alditol is a residue of tritol, tetritol, pentitol or hexitol. 20. The method according to any one of claims 1 to 8 for the production of nitrosourea derivatives of the formula q (form lh) NO A5 NHCN ^ (le) «" ° / no 21. The method according to any one of claims 1 to 8 for the production of nitrosourea derivatives of the formula 619 967 22. The method according to any one of claims 1 to 8 for the production of nitrosourea derivatives of the formula 23. The method according to any one of claims 1 to 8 for the preparation of l-ethyl-3- (ß-D-glucopyranosyl) -l-nitroso-urea, 1 - (ß-D-glucopyranosyl) -3-nitroso-3-n -propylurea, 1-n-butyl-3- (ß-D-glucopyranosyI) -l -nitrosourea or 1 - (2-chloroethyl) -3- (ß-D-glucopyranosyl) -l -nitrosourea. 24. The method according to any one of claims 1 to 8 for the preparation of l- (2-chloroethyl) -3- (ß-D-mannopyranosyl) -l-nitrosourea or l- (2-chloroethyl) -3- (ß-D- galactopyra-nosyl) -1-nitrosourea. 25. The method according to any one of claims 1 to 8 for the preparation of l-methyl-l-nitroso-3- (ß-D-xylopyranosyl) urea, l- (2-chloroethyl) -l-nitroso-3- (ß- D-xylopyranosyl) urea, 1 - (1,3 / 2N-dihydroxycyclohexyl) -3-methyl-3-nitro-sourea or l- (2-chloroethyl) -3- (1,3 / 2N-dihydroxycyclo-hexyl) -1-nitrosourea. 26. The method according to any one of claims 1 to 8 for the preparation of 1 - (ß-D-mannopyranosyl) -3-methyl-1-nitrosourea, methyl-2,6-dideoxy-2- (N'-methyl-N'- nitrosourei-do) -ß-D-glucopyranoside or methyl-2-deoxy-6,0-mesyl-2- (N'-methyl-N'-nitrosoureido) -ß-D-glucopyranoside. 27. The method according to any one of claims 1 to 8 for the preparation of l- (2-chloroethyl) -l-nitroso-3- (lN, 3,5 / 2,4,6-pentahydroxycyclohexyl) urea. 28. The method according to any one of claims 1 to 8 for the production of l- (β-D-maltosyl) -3-methyl-3-nitrosourea, l- (2-chloroethyl) -3- (β-maltosyl) -l-nitrosourea , l- (β-D-lactosyl) -3-methyl-3-nitrosourea or l- (2-chloroethyl) -3- (β-D-lactosyl) -l-nitrosourea. 29. The method according to any one of claims 1 to 8 for the preparation of N-carbamoyl-N '- (2-chloroethyl) -N'-nitroso-D-glucosaminol or N-carbamoyl-N' - (2-chloroethyl) -N ' -nitroso-D-galactosaminol. 30. The method according to any one of claims 1 to 8 for the preparation of (N-carbamoyl-N'-methyl-N'-nitroso) - (O-carbamoyl-N-methyl) -ethanolamine or [N-carbamoyl-N '- ( 2- 31. The method according to any one of claims 1 to 8 for the preparation of l- (6-deoxy-ß-D-glucopyranosyl-3-methyl) 3-nitrosourea or 1 - (2-chloroethyl) -3- (6-deoxy-ß -D-gluco-pyranosyl) -1 -nitrosourea. 32. The method according to any one of claims 1 to 8 for the preparation of tetra-N- (N'-methyl-N'-nitroso) carbamoy) -ribostamvcin or tetra-N- [N '- (2-chloroethyl) -N'- nitroso] carbamoyl ribostamycin. 33. Compound of the formula I, prepared by the process according to claim 1. 34. Connection according to claim 33, produced according to one of claims 2 to 32. 0 ! J nhcn -no • rj (Ia)