FI65782C - ANALOGIFICATION OF THE NUTRITIONAL CARBIDE OF N1-GLUCOFURANOSID-6-YL-N3 - Google Patents

Description

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(32) (33) (31) Pretty · ακ> <Ι («ι · -Β · | Μ frlorkat 21.02.78 Switzerland-Switzerland (CH) 1849 / 78-5 (71) Ciba-Geigy AG, CH-4002 Basel, Switzerland-Schweiz (CH) (72) Jaroslav Stanek, Birsfelden, Switzerland-Schweiz (CH) (74) Oy Jalo Ant-Wuorinen Ab (54) Analogue method for the preparation of therapeutically active N-glucofuranosid-6-yl-N-nitrosoureas This invention relates to an analogous process for the preparation of therapeutically active N, N-glucofuranosid-6-yl N, N-nitrosoureas of the compounds of the formula: N-glucofuranosid-6-yl-N-nitrosocarbamide

WELL

CH2 - NH - CO - N - R6 R - O - CH

O-R 2 wherein lower alkyl, phenyl lower alkyl or Lower alkanoyl, R 2 is hydrogen, lower alkyl or lower alkanoyl, R 1 and R 2 together are lower alkylidene, R 5 is hydrogen, lower alkyl, phenylalkylalkyl or lower alkanoyl optionally halogen, and R

In the following, the following residues, radicals or compounds with the word "lower" contain, unless specifically mentioned, 2 65782 preferably up to 7, in particular up to 4 carbon atoms.

Lower alkyl is, for example, isopropyl, straight-chain or branched, butyl, pentyl, hexyl or heptyl, and preferably methyl, ethyl or n-propyl.

The substituted lower alkyl residue may contain one, two or more halogen atoms.

Particularly mentionable as phenyl-lower alkyl are 2-phenylethyl, chlorobenzyl, methylbenzyl, hydroxybenzyl, methoxybenzyl or especially benzyl.

The lower alkylidene residue is, for example, a 2-butylidene, 3-pentylidene and, in particular, an isopropylidene residue.

Acyl is containing from 2 to 18 carbon atoms, such as acetyl or propionyl.

Halogen is, for example, fluorine, chlorine or bromine.

The new compounds may be in the form of a mixture of anomers or as pure α- or β-anomers.

The new compounds have valuable pharmacological properties, in particular they have a very good effect on various tissue transplant tumors and leukemia.

They are effective at doses ranging from 25 to 500 mg / kg i.p. strongly inhibiting tumor growth in mice with Ehrlich-Ascites cancer and in rats with e.g. Walker CaSa 256. Corresponding doses have a prolonging effect on life expectancy when observed in mice with e.g. Leukemia L 1210.

Thus, for example, ethyl 6-deoxy-5-O-methyl-6- (3-methyl-3-nitrosourea) -β-D-glucofuranoside 50-250 mg / kg in ip doses of 80-100% was obtained. The growth inhibition and leukemia of the above-mentioned tumors in L 1210 prolonged the age by more than 90%. Similar results were also obtained with oral dosing. Suitability is good. There are no side effects even during long-term treatment. Also, no tissue changes were observed macroscopically in normal animals after three weeks of oral administration.

In C.A. 8j5, 16911j describe antitumor 5- (nitrosoureido) -5-deoxy-D-ribofuranose derivatives. Compounds of this type differ in structure from the compounds of this application in that they have a ribofuranose backbone while the compounds of the invention have a glucofuranose ring in place of them.

3,65782

However, methyl 5-deoxy-2,3-O-isopropylidene-5- (3-methyl-3-nitrosoureido) -g-D-ribofuranoside obtained according to the cited literature has been used as a known reference II in the present experimental description.

In J.Med.Chem. 1_7, 392-396 (1974) describe streptosotocin analogs for use in the treatment of leukemia. More specifically, it deals with 3-methyl-3-nitrosoureido derivatives of amino sugars, as further defined on page 393. On page 395, in the right-hand column, 3-deoxy-1,2-O-isopropylidene-3- (3-methyl-3-nitrosourureido) -α-D-ribofuranose, which is structurally very close to the above, is described in Experimental Report as Compound II methyl 5-deoxy-2,3-O-isopropylidene-5- (3-methyl-3-nitrosoureido) -β-D-ribofuranoside, so that it seems reasonable to assume that the compounds mentioned in the literature cited have essentially the same properties as test report with the corresponding compound II. Thus, the inclusion of the compound described in said literature reference in the experimental report does not seem necessary, however, the literature referred to has been sufficiently taken into account in the prior art, especially given that the amino sugars described therein differ too much from the glucofuranose form of the compounds of the invention.

German Patent Publication No. 25 30 416 describes alkyl deoxy- (3- (2-chloroethyl) -3-nitrosourea) -D-glucopyranoside with potential antitumor activity and the general formulas defined therein. Because it deals with compounds based on the D-glucopyranose backbone, such compounds differ in structure from the D-glucofuranose-type compounds of the invention, so that no direct comparison has been made between the two classes of compounds.

However, reference is made here to what has become known as CHLOROSOTOSINE and in J.Med.Chem. 104-106 (1,075) tumors to 2- (3- (2-chloroethyl) -3-nitrosourea) -2-deoxy-D-glucopyranose described as a resistance compound, which differs from the compound described in German Patent Publication 25 30 416 only in that: that in the formula Ia R is hydrogen. The attached test report includes CHLORO-SOTOSINE as the reference compound. Given the structural similarity between CHLOROSOTOSINE and the compound described in German Patent 25 30 416, it can be reasonably assumed that there is no significant difference in effect between the two classes of compounds, so that the inclusion of the compounds described in the referenced German patent does not appear necessary.

4,65782

The test report

The following compounds have been tested for their antitumor and anti-leukemic activity: I. ethyl-5-O-methyl-6-deoxy-6- (3-methyl-3-ureido) -α-D-glucofuranoside; II. methyl-5-desoxy-2.3-0-isopropylidene-5- (3-methyl-3-nitroso-ureido) -3-D-ribofuranoside; (known Chemical Abstracts, 86, 1 6911 j (1977)).

III. 2- (3- (2-chloroethyl) -3-nitrosoureido) -2-deoxy-D-glucopyranose-si; (known as J. Med. Chem. 1J), 1975, 104-106 and as CHLOROSO-TOSINE).

A. Experimental Methods 1. LD4: Means the dose at which 10% of the experimental animals (mice) died within 7 days of the start of the experiment.

2. B16-melanoma

Groups of ten female mice, strain C57 / BLf / BOM (SPF) C, weighing 18-22 g, were administered s.c. or i.m. 0.5 ml of solid tumor suspension in Hank's solution. Four hours after the transplant, treatment was started with Compound I 250 mg / kg 15 times (over three weeks, five times a week) po, Compound II 764 mg / kg once (= LD10) and ten times 191 mg / kg ten times. kg (= 1/4 LD ^), over two weeks (five times a week po, and once 313 mg / kg (= LD.jg) 3a I go 78 mg / kg (= 1/4 LD.jg) ip · two weeks (five times a week), and Compound III 258 mg / kg (= LD ^ g) once and 64.5 mg / kg (= 1/4 LE> -qq) ten times per week) po, and once 27 mg / kg (= LD · ^ fcyroroe1'1®1'1 times 6.75 mg / kg (= 1/4 LD1Q) ip over two weeks (five times per week). After 22 days The tumor weight of both the treated animals and the untreated control animals was determined to record the results. n tumor weights were performed by the methods described by Bartlett1 and Dunnetf.

3. Leukemia (6-mercaptopurine-sensitive)

Female mice (strain B ^ ^ F ^) were inoculated with 10® Leu chemistry cells in each case. At four, 5 6 5 7 8 2 24: nf 48 and 72 hours, respectively, a group of ten animals was administered the test compound orally (p.o.) or intraperitoneally (i.p.), and the animals were observed for the following days. Untreated control animals died after 7-8 days. The increase in the average lifespan of the treated animals was compared with the lifespan of the control animals and expressed as a percentage.

B. Table of combined LD50 q mg / kg B16 melanocma: Leukemia L 1210 / S2: te mg / kg Tumor mg / kg Decreased lifetime weight compared to untreated untreated controls to these roll animals% ___% _ I> 2500 mg / kg po 15 times 4 times 250 p.o. 28 1250 mg / kg i.p. 250 p.o. 4 times 125 p.o. 14 4 times 250 i.p. 75 4 times 125 i.p. 57 II 764 mg / kg p.o. once all animal- once 764 p.o. 14,313 mg / kg i.p. 764 p.o. met died 10 times 21 times 382 p.o. 3,191 p.o. c once 3 times 313 i.p. 13 131 i.p. once 156.5 i.p. 10 10 times 78 i.p. 13 III 258 mg / kg p.o. once 27 mg / kg i.p. 258 p.o. 10 times 258 p.o. 15 10 times 64.5 p.o. 53 times 129 p.o. 19 times 27 i.p. 72 times 27 i.p. 161 10 times all animal 6.75 i.p. met once died 13.5 179 65782 C. Conclusions

In terms of toxicity, Compound I has been shown to be at least three times less toxic than Compound II and at least ten times less toxic than Compound III when administered orally. When administered intraperitoneally, Compound I is about four times less toxic than Compound II and about 50 times less toxic than Compound III.

Its anti-B16 melanoma activity was found to be about three times more potent than compound II when administered orally, while the effect of compound II when administered intraperitoneally was negligible. Compared to Compound III, Compound I is clearly superior when administered orally. Compound I is at least about four times more potent than Compound II in its anti-mercaptopurine-sensitive leukemia L 1210 / S2 activity, whereas Compound I is only slightly more potent than Compound III when administered orally, and Compound I is clearly superior to Compound III when administered intraperitoneally. Given that compound II, and in particular compound III, is much more toxic than compound I, the latter is superior to compounds II and III.

Preference should be given to compounds in which R5 is methyl,

R 9 is methyl or chloroethyl and R 1 is hydrogen, methyl, ethyl or propyl and R 2 is hydrogen or R 1 and 3a R 2 together form an isopropylidene residue.

The new compounds are obtained when the compound of formula II is CH2 - NH - CO - NH - R6 R5 - 0 - (II)

Kp o - R2 are nitrosated in a manner known per se.

In this case, the compound of the formula II is preferably reacted with nitric acid, its salts or derivatives. In this case, it is preferable to use a salt of nitric acid, such as an alkali or alkaline earth metal salt, in particular a sodium salt, and then to liberate the nitric acid from an acid such as a mineral acid, e.g. hydrochloric acid, sulfuric acid or nitric acid, an organic acid such as carbonic acid, acetic acid. or a sulfonic acid, e.g. a lower alkyl sulfonic acid, such as methane or ethanesulfonic acid, or an ion exchanger containing sulfonic acid groups, e.g. Amberlite IR 120. It is also possible to use nitric anhydride, in particular a mixed anhydride formed, for example, with nitric acid or hydrohalic acid.

If necessary, work is carried out in the presence of a solvent, in which case, for example, the present organic acid can also be used as such. The reaction is preferably carried out at a low temperature, e.g. -10 ° to + 30 °.

The starting materials used in these production methods are new. They are prepared in a manner known per se from the corresponding 6-position-unsubstituted glucofuranose, e.g. by reacting it, e.g. with alkanesulphonic acid, arylsulphonic acid or hydrohalic acid, to give a reactive ester, then forming the azide and reducing the azide 6-glucose 6-desoxy thus obtained. , which is then condensed with a suitable N-Rg-carbamic acid derivative such as the corresponding isocyanate to give 6-deoxy-6- (3-Rg-ureido) -glucofuranose. As mentioned above, these reactions are carried out in a manner known per se.

Another process for the preparation of novel nitrosoureas comprises the compound of formula III

CH2 - NH2 R5 - 0 - CH

- Rx (III) 0 - Ή yr o - r2 is reacted with a compound of formula IV

N - nitroso - N - Rg - carbamic acid IV

with a reactive derivative.

8 65782

The reactive derivative may be, for example, an acid anhydride, preferably a mixed acid anhydride such as an acid azide or an activated ester. Activated esters which may be mentioned are, in particular, cyanomethyl ester, carboxymethyl ester, paranitrophenylthioester, paranitrophenyl ester, 2,4,5-trichlorophenyl ester, pentachlorophenyl ester, N-hydroxysuccinimide ester, N-hydroxyphthalimide ester, 8-hydroxyquinoline

These reactions are carried out in a manner known per se, preferably in a solvent such as water, a halogenated hydrocarbon, e.g. dichloro- or trichloroethane, an ether such as diethyl ether, tetrahydrofuran, dimethylformamide, dimethylsulfoxide or optionally alkylated pyridine, picoline, pyridine, such as pyridine. .

The starting materials used are known or are prepared in a manner known per se. The amine of formula III can be obtained from the corresponding 6-unsubstituted glucofuranose, e.g. by reacting it, e.g. with time or arylsulphonic acid or hydrohalic acid, to give a reactive ester, then converting it to the azide and reducing the azide thus obtained to 6-deoxy. 6-amino-glucofuranose.

The above processes are carried out by methods known per se, without or preferably in the presence of diluents or solvents, if necessary with cooling or heating, under elevated pressure and / or in an inert atmosphere, e.g. a nitrogen atmosphere.

The new compounds may exist as pure α- or δ-anomers or as mixtures of anomers. The latter can be separated into both pure anomers in a manner known per se on the basis of the physicochemical differences of the components, e.g. using chromatographic separation such as thin layer chromatography or some other suitable separation method. Preferably, the more efficient one is isolated from the two anomers.

The invention also relates to embodiments of the process in which the starting material is formed under reaction conditions or the starting material is used in the form of a reactive derivative or salt. In this case, the starting materials are preferably used which, according to the process, give the compounds mentioned above as being particularly valuable.

9 65782

The following examples illustrate the invention described above; however, they do not limit the invention in any way. Temperatures are expressed in degrees Celsius.

Example 1;

To a solution cooled to 0 ° containing 13.5 g of ethyl 5-deoxy-5-O-methyl-6- (3-methylureido) -D-glucofuranoside in 150 ml of distilled water and 6, 2 ml of glacial acetic acid, a solution of 3.6 g of sodium nitrite in 20 ml of distilled water is added dropwise over 2 hours. Stir this reaction mixture for 16 hours in an ice bath, saturate with sodium chloride and extract five times with 200 ml of chloroform. The chloroform extracts are combined, dried over sodium sulfate and evaporated. The residue is dissolved in 100 ml of distilled water and dried by cooling. Ethyl 6-deoxy-5-O-methyl-6- (3-methyl-3-nitrosoureido) -α-D-glucofuranoside is obtained, m.p. is 54-55 ° and (a)? = + 1 ° (chloroform, c = 1.003).

The starting material can be prepared as follows:

To a solution of 52 g of 6-azido-3-O-benzyl-6-deoxy-1,2-O-isopropylidene-α-D-glucofuranose in 600 ml of N, N-dimethylformamide is added 8 g of sodium hydride pract. (oil-free), stir for one hour at room temperature and cool to 0 °. 19.2 ml of methyl iodide are then added dropwise over 30 minutes and the mixture is further stirred for one hour. Filter and evaporate the solution. Distilled water is added to the residue and it is extracted with ether. The ether solution is washed with distilled water until neutral, dried over magnesium sulfate and evaporated. 6-Azido-3-O-benzyl-6-deoxy-1,2-O-isopropylidene-5-O-methyl-αD-glucofuranose is obtained, which is a yellowish oil with (a) ^ = -59 ° + 1 ° (chloroform, c = 0.667).

A solution of 54.6 g of 6-azido-3-O-benzyl-6-deoxy-1,2-O-isopropylidene-5-O-methyl-α-D-glucofuranose in 430 ml of abs. ethanol and 120 ml of 4.6N alcoholic hydrochloric acid are left to stand at room temperature for 18 hours and evaporated in a water-jet vacuum. The residue is dissolved in ether, washed with saturated sodium hydrogen carbonate solution and distilled water, dried over magnesium sulphate and evaporated. Column chromatography, using 1.2 kg of silica gel, eluting with methylene chloride / ethyl acetate 19/1, gives ethyl 6-azido-3-O-benzyl-6-deoxy-656582 5-O-methyl-α-glucofuranoside, which is colorless oil with (α) ρ ° = + 35 ° + 1 ° (chloroform, c = 0.834).

10.1 g of ethyl 6-azido-3-O-benzyl-6-deoxy-5-O-methyl-α-glucofuranoside in 100 ml of ethanol are reduced, in the presence of 0.5 g of 5% palladium / carbon, with hydrogen Within 90 minutes. After filtering off the catalyst, 36.7 ml of methyl isocyanate are added dropwise to the filtrate, while stirring and cooling externally, and evaporated to dryness. The residue is crystallized from ethyl acetate / ether. Ethyl 3-O-benzyl-6-deoxy-5-O-methyl-6- (3-methylureido) -α-D-glucofuranoside is obtained, m.p. 92-93.5 ° and (a) ^ = + 42 ° + 1 ° (chloroform, c = 0.973).

This product is dissolved in methanol and hydrogenated in the presence of 10% palladium / carbon. At the end of the work-up, ethyl 6-deoxy-5-O-methyl-6- (3-methylureido) -α-D-glucofuranoside is obtained, which is an oil with = + 73 ° + 1 ° (chloroform, c = 0.330).

Example 2:

To a solution of 4.7 g of ethyl 6-amino-6-deoxy-5-O-methyl-α-glucofuranoside in 40 ml of chloroform, cooled to 0 °, is added dropwise, with stirring, a solution of is 2.5 g of N-nitrosomethylcarbamyl azide in 40 ml of ether and is further stirred for 3 hours at room temperature. This solution is evaporated to dryness and the residue is dissolved in distilled water. Extract once with ether, separate the aqueous phase and lyophilize. Ethyl 6-deoxy-5-O-methyl-6- (3-methyl-3-nitrosourea) -α-D-glucofuranoside is obtained, m.p. is 54-55 ° and (a) ^ = + 65 ° + 1 ° (chloroform, c = 0.914).

Example 3:

In a similar manner, starting from the corresponding starting materials, the following compounds were prepared: a) ethyl 6- (3- (2-chloroethyl) -3-nitrosoureido) -6-deoxy-5-O-methyl-α-glucofuranoside, (α) ο ° = + 53 ° ± 1 ° (chloroform, c = 1.231), b) ethyl 6-deoxy-6- (3-methyl-3-nitrosoureido) -D-glucofuranoside, (a) ^ = + 31 ° ± 1 ° (water, c = 1.124), c) 6-deoxy-1,2-O-isopropylidene-6- (3-methyl-3-nitroso-ureido) -crD-glucofuranose, m.p. 115 ° (dec.), (U) q ° = -11 ° '1 ° (chloroform, ^ ο = d) ethyl-5-O-ethyl-6-deoxy-6- (3-methyl-3-nitrosoureido) - uD-glucofuranoside, fa) + = 57 ° (1 ° (chloroform, c - 1.023), 11 65782 e) ethyl-6-deoxy-6- (3-methyl-3-nitrosoureido) -5-0 - propyl-α-D-glucofuranoside, (a) = -45 ° ± 1 ° (chloroform, c = 1,045), f) ethyl-5-O-benzyl-6-deoxy-6- (3-methyl- 3-nitrosoureido) -D-glucofuranoside, (a) = + 41 ° ± 1 ° (chloroform, c = 1.217), g) methyl-6-deoxy-6-p-methyl-S-nitrosoureido) -5- O-methyl-α-glucofuranoside, m.p. 82-83 °, (a) 76 ° ± 1 ° (chloroform, c = 1.001), h) ethyl 6-deoxy-2,5-dimethyl-O-methyl-6- (3-methyl-3-nitroso- ureido) -D-glucofuranoside, m.p. 93.5-95 °, (a) ^ ° = + 97 ° + 1 ° (chloroform, c = υ 1.049), i) ethyl-2-O-acetyl-6-deoxy-5-O-methyl-6- (3-methyl-3-nitrosoureido) -α-glucofuranoside, m.p. 55-57 °, (a) 3 ° = + 109 ° + 1 ° (chloroform, c = 1.01), j) 6-deoxy-1,2-di-O-acetyl-5-O-methyl-6 - (3-methyl-3-nitrosoureido) -D-glucofuranose, (a)? = = 17 ° + 1 ° (chloroform, c = 0.995), k) benzyl-6-deoxy-5-O-methyl-6 - (3-methyl-3-nitrosourea) -D-glucofuranoside, (α) D = + 69 ° + 1 ° (chloroform, c = 1,318), 1) 5-acetyl-6-deoxy-1,2- O-isopropylidene-6- (3-methyl-3-nitrosoureido) -α-D-glucofuranose, (α) D = -26 ° + 1 ° (chloroform, c = 1.107).

Claims (2)

1. Analogous Process for the Preparation of Therapeutically N-glucofuranosid-6-yl-N, -nitrosocarbamides of Formula 100 ip CH - ri <*> * • N. 0 - H / 1 R 2 in which R 2 represents lower alkyl, phenyl lower alkyl or lower alkanoyl, R 2 hydrogen, lower alkyl or lower alkanoyl, R , that a) a compound of the formula CH