AU602763B2 - Nitrosourea derivatives, a novel procedure for their preparation and their therapeutic applications - Google Patents

Description

Fo~rm COMMONWEALTH OF AUSTRALIA PATENTS ACT 1952.69 COMPLETE SPEC IFICATION

(ORIGINAL)

Class Int. Class Application Number: Lodged: 15109/88 22.04.1988 Complete Specification Lodged: Accepted: Published: Friority: Related Art: Nzime of Applicant: SANOFI 40 Avenue Ceorge-V, 75008 Paris, France Ai :ress of Applicant Actual lnventort PIERRE ROGER, JEAN-PAUL FOURNIER, CLAUDE MONNERET ALPIN MARTIN and Address for Service EDWD, WATERS SONS, 50 QUEEN STREET, MELBOUJRNE, AUSTRALIA, 3000, Complete Specification for the invention entitled: NITROSOUREA DERIVATIVES, A NOVEL PROCEDURE FOR THEIR PREPARATION AND THEIR THERAPEUTIC APPLICATIONS The frnllowing statement is a full description of this invention, including the hest method of parl'orming it known to u NITROSOUREA DERIVATIVES, A NOVEL PROCEDURE FOR THEIR PREPARATION AND THEIR THERAPEUTIC APPLICATIONS.

The present invention relates to new nitrosourea derivatives, and more especially to novel 4-deoxy sugar derivatives of nitrosoureas, to a new procedure for their preparation and to their uses as therapeutically active substances.

2-aeoxy sugar derivatives of nitrosoureas and 4-deoxy sugar derivatives of nitrosoureas have already been described in the French patent application No. 83.13878 filed on 30 August 1983 (published under .the No. 2 551 068); these compFounds exhibit useful therapeutic properties, notably antitumor properties.

More precisely, the above mentioned patent application describes compounds correspondin" to the following general formula A 0

A

X

R,

in which -R represents a hydrogen atom, an alkyl group trrml to 30, preferably 1 to 12 carbon atoms or an aralkyl 1_14 group frti~7 to 12, preferably 7 to 9 carbon atoms, optionally substituted by one or several, particularly up to 3, halogen atoms, NO 2 NH2, CF 3 groups or alkoxy groups of 1 to 4 carbon atoms, -X represents a hydroxy group or an NIRR 2 group -Y represents a hydrogen atom a hydroxy group or an NR' 1 group 2 where R 1 and/or R' each represent a hydrogen atom or a -C-N-CH2CIIHal group, Hal being a halcgen, preferably Cl, it1 2 2 0 NO and R 2 and/or H' 2 each represent a hydrogen atom, an alkyl I group comprising from 1 to b carbon atoms, an aralkyl group comprising 7 to 12, preferably 7 to 9 carbon atoms, a cycloalkyl group comprising from 3 to b carbon atoms, an aryl group of 4 to 10 carbon atoms, the aryl and aralkyl groups being possibly substituted by one or several, particularly up to 3, halogen atoms, NO 2

,NH

2

,CF

3 groups or alkoxy groups of 1 to 4 carbon atoms, R' and R" represent hydrogen, OM, M representing an alkyl group comprising from 1 to 30, preferably from i to 12 carbon atoms, an aryl group frn 4 to 10 carbon atoms, anx aralkyl group comprising from 7 to 12, preferably from 7 to 9 carbon atoms, the aryl and aralkyl groups being possibly substituted by 1 or several, particularly up to 3, halogen atoms, NO2,NH 2

,CF

3 groups or alkoxy groupsfran 1 to 4 carbon atoms, or M represent ina in acyl group frcm 2 to 8 carbon atoms, preferably 2 or 3,or an arbylgroup firc 5 to 12, preferably 5 to 9 carbon atoms, unsubstituted or substituted by one or several, particularly up to 3, NO 2

NH

2

CF

3 groups, halogen, alkoxy of 1 tc 4 carbon atoms, provided that either R' or P" represents hydrogen, R' and R" being not simultaneously hydrogen and

R

provided that at least X represents -H I ,with R 2 I with representing -C-N-CH 2 CH2Hal, or Y represents -N wit \p 0 NO 2 R'I representing -C-N-CH 2

CH

2 Hal 0 NO 3 These compounds are obtained from the compound of formula A bis C112 Y OR A bis x in which X1 represents a hydroxy or -NHR 2 group IS- Y1 represents a )-l-gen atom, hydroxy group, or -NIR 2 group; R, R I, R 2 anu. have the above-indicated meanings In formula A, and at least one of the groups XV or Y' represents the NBR 2 or N1IR' 2 rroup 1) which compound i8 reavctro with a 2-ethyl-halogeno isocyanate, to convert the KHR 2 or NHfl 2 group of the compound of forlnulaA bis respectively

NR

2

NHOH

2

CHI

2 Hal or NR' NHCH 2 ,CH.,Hal Hal being a halogen atoms, 2) the compound obtained in the preceding'step Is subjected to nitrosation, by means of a nitrxite of an alk~ali metal, to convert theI N'R CNHCH CH Hal or NR' 2 CNHCH CH Hal groups respectively 0 0 into NR2 C- C 2 CH 2 Hal or NR 2 C-N-C-1 2

CHI

2 MaL.

The compounds of formula A bis are obtained from intermediary compounds comprising a primray amine grou~p In position 3 and/o~r 6, as follows:

CH

2 Y' (or NH 2 CH 2 Y' (or NHR' 2 0 R CH 0 OR 2R' OR .reducing agent ou IR 1 Ha1 COR Rl" X (or NH 2 R 2 X reducing agent (orNHR 2 In the case of desoxy-2 compounds of D configuration, these intermediary compounds comprising a primary amine can be obtained, starting from azide comprising the desoxy-2 function with a hydrogen in position 2.

In the case of desoxy-2 of P-L configuration, the compounds comprising the primary arine can be obtained, starting from the azide comprising the desoxy-2 function with a hydrogen in position 2.

The desoxy-4 compounds in which Y represents hydrogen can also be obtained from equivalent processes starting either from the azide comprising the desoxy-4 function with a hydrogen in position 4, for example for the desoxy-4 compounds of P-L configuration, or from an intermediary compound comprising a primary amine group, for example for the desoxy-4 compound of a-D configuration.

However, it happens that the access to some Ij nitrosoureas could be encompassed only by other routes, specially for the synthesis of starting material or for the synthesis of intermediary products. This is particularly the case for the compounds of formula A in which R' represents hydrogen and Y represents hydroxy, for which the compounds with an amine function or an azide group comprising the desoxy-4 function with the hydrogen in position 4.

As a result, the desoxy-4 compounds with a D configurat on are a special case and consequently an original class of compounds which the present invention now makes available. This is as remarkable as: some members of this particular class of compounds present anti-tumor activities which have never been reached so far in pharmacological tests which are considered as particularly significant today; the process hereafter described provide means to one skilled in the art tco othr classes zf moepcunds, among which some present anti-tumor activities which are also as remarkable: it is particularly the case of the compounds in which the carbon in position 4 can be substituted by a halogen at"m and 'r the carbon in position 6 can be substituted by an azyl gt up.

The subject of the invention consists of new cntrosourea derivatives characterized in that they correspond to compounds with the formula I below:

CH

2

R

6

OI

HS OR 1

OH

in the form of one of the two anomers, 0( or in which: 1 represents an alkyl group of 1 to 12 carbon atoms, or ar aralkyl group of 7 to 12, preferably 7 to 9, carbon atoms, possibly substituted in the aromatic nucleus by 1 or more, in particular up to 3, halogen atoms, by 1 to 3 NO 2 or CF 3 groups or alkoxy groups containing 1 to 4 carbon atoms, S R represents H or Hal, IHa! being a Cl or Br atom, especially Cl, 6

R

6 represents OH or O-C-R, R corresponding to nalkyl group of 1 to 6 carbon atoms, an aryl group,unsubstituted or substituted on the aromatic nucleus by 1 or more atoms, in particular up to 3 halogen atoms, by 1 to 3 NO 2 or CF groups or alkoxy groups of 1 to 4 carbon atoms.

Nu represents the group NH-C-N-CH 2

-CH

2 -Hal Hal' corresponding to a halogen chosen from among F, C1, Br and I, identical with, or different from Hal and being in particular chlorine.

It has been observed that the new compounds thus obtained exhibit a better therapeutic index, considered generally and globally, than that for all known such compounds, thus making these compounds without equivalent among those of the family to which they belong Sthat the new 4-deoxy sugar derivatives of nitrosoureas exhibit physico-chemical properties which facilitate their therapeutic use; that the new 4-deoxy sugar derivatives of nitrosoureas are available in a solid, stable formi Sthat the new 4-deoxy sugar derivatives of nitrosoureas possess remarkable activity towards tumors which are usually very difficult to treat and, in particular making it possible to bring about the regression of certain types of tumors -that the rew ,ompounds are therapeu- .tically active, the activity of which is foaid to be equally efficacious when these new compos.ds are administered by the intraperitoneal route to the animal as when they are administered by the intravenous route, a circumstance which is not usully obtained with this type of compound, and is predkctive of an activity iA man, 4 7 In formula I and some of the formulae presented later the bond between the hydrogen atom and OR 1 group at position 1 of the sugar ring is represented by the symbol IH,OR This representation signifies that the OR I group can be either in the t position or in the position according to the HAYWORTH representation.

In the remainder of the description the term alkyl includes linear, branched or cyclic (cycloalkyl) alkyl groups.

The substances according to the invention represented by formula I are 3,4-dideoxy v- -D-xylohexopyranosides, 3,4-dide'xyv -D-xylohexopyranosides, 3,4-dideoxy-O( -D-galactohexopyranosides and 3,4-dideoxy-P D-galactohexopyranosides.

An advantageous class of compounds according to the invention is constituted by the nitrosourea derivatives corresponding to the formula I above and in which:

R

1 represents an alkyl group of 1 to 12 carbon atoms,

R

4 represents a halogen atom,

R

6 represents an OH or O-C-R, R having the meanings indicated above, In this class of nitrosourea derivatives acccrdlng to the invention particularly advantageous compounds of formula I indicated above are those in which: R I represents a CH 3 group, R represents a halogen atom, R represents OH or O-C-R, R having the meanings indicated above.

'I

L ~i N8 In this same class of nitrosourea derivatives according to the invention particularly advantageous substances of formula I indicated above are those in which, on the one hand:

R

1 represents a CH 3 group, R represents Cl,

R

6 represents OH or O-Q -RR having the meanings indicated above and, on the other:

R

1 represents a CH 3 group, R represents a halogen, 0-C-R

R

6 represents b ,R having the meanings indicated above.

Another advantageous class of nitrosourea derivatives according to the invention is constituted by those corresponding to formula I indicated above in which: R represents a CH 3 group,

R

4 represents a hydrogen atom,

R

6 represents

OH.

Another advantageous class of nitrosourea derivatives according to the invention is constituted by those correspondinr to formula I indicated above in which: R represents a CH 3 group, R represents a hydrogen atom,

R

6 represents O-fR, R having the meanings indicated above.

Among the compounds cited above, the most interesting are those having the following formulae: I' 1 ?0

CH

9 0H CH OH 1803 1676 1674 1675 '167 7 OCH 3 The present invention also relates to a procedure for the preparation of the new derivatives corresponding 'o formula I: C26 H,

ORI

in the form of one of the two anomers, c4 or in which RI represents an alkyl group of 1 to 12 carbon atoms, or an aralkyl group of 7 to 12, preferably 7 to 9, carbon atoms, possibly substituted by I, or more, in partic'l~ar up to 3, halogens ato-s, by I to 3 NO 2 or Cgroups or alkoxy groups of I to 4 carbon atoms, R, represents Hal, Hal being a halogen atom, in particular Cl, rrppresents OH or O-C-R, R having the meanings indicated above.

0 Nu represents the group ,Hal' being identical with, or different from Hal and representing a halogen atom, in particular ohlorine, characterized in that a) in a first series of rei-ctions the compound represented by formula II CH 0" 0

O(H

Oil is made to react in the form of' a mixture of the and anomors with a glycosylating agent constituted by an alcohol R 1 OHI Al having the meaning indicated above in order to convert the OH group ir position I into the

OR

I group, Sthe product thus obtained is treated with an activated derivative of the acid R-COOH in order to convert the OH group in position 6 into the O-C-R group, 0 the 9 and anomers are separated by a suitable method and 'each of the anomers, or is made to react with a halogenating agent in order tp introduce a halogen atom at vositon 4, the compound obtained being present as one of the anomers K errand S corresponding to the formula III, CH,0 0 0 lIol Hal 0 N 3 H, OR III

OH

in which S R ani R 1 have the meanings indicated in formula I, Hal represents a halogen atom b) subsequently, in a second step, the compound represented by formula III is subjected to a reduction in order to reduce N 3 to NH 2 thi.

3 E reduction being carried out, in addition, under conditions such that the halogen atom in position 4 may or may not be hydrogenolyzed, this reduction being possibly followed by hydrolysis of the O-C-R 0 group in position 6 in order to generate an OH group, the compound obtained ort completion ot this Second step being pre4 1 t in the form of one of the anomers o( or P and corresponding to the formula IV below I 12 CH R 6 R 0

IV

-NH 2 H, OR 1

OH

Rl and R6have the meanings indicated in formula I c) in a third series, of reactions the anomers/ d. or represented by formula IV 4re sbjected to reaction with a halogeno ethyl isocyanate in ord toQover N ino HCNHCH CH Hal 'and the prodict thus ord~r;o~over N 2 nt N11 2 2 0 [obtained is treated with the nitrite of an alkali metal, in particular isium nitrite, in order to convert NH ~NHCH:2CH:2Hal' into

NHCNCH

2

CH

2 "al' ,in order to obtain the compouind represented by Inth ormula It and some of the subsequent formulaeA the bond between the OH and the carbon atom at position 2 of the cyclic structure Is represented~ by the symbol r'V This representation signifies that the OH group can be in either the b position or the Position.

As activate, derivative of the acid R-COOH,use may made of the ac.~d derivatives used for the estorification of the primary hydroxyl.

~groups, such as acid halide, preferably chloride, or anhydride, As halogenating agentisulfuryl chloride or bromide is, preferably used.

The starting material corresponds to formula 11 H 2

ON

0\~O v

OH

in the form of a mixture of the ind 7 anoners ,s described in the literature (particularly by Redlich in Liebigs Chem. 1981, p. 1215 1222).

A preferred embodiment of the procedure of the invention for the preparation of the nitrosourea derivatives corresponding to formula I comprises the reaction of the compound correspunding to formula II above in the form of a mixture of the c. and P anomers with a glycosylating agent constituted by R OH, R 1 having the meaning indicated above in order to lead to the formation of the compound corresponding to the formula V.

CH

2 0H 0. V the introduction of an acyloxy group, of the formula -C-R at position 6, R having the meaning indicated above in order to give rise to the compound with the formula VI the separation of the compound VI into the two anomers, V and corresponding to the formula VII i i i F_ 1 CH 2 R H, OR1 -the reaction of the anomers, rj4 or P corresponding to formula VII, with an halogeniating agent in order to give rise to the anomers U.or having the formula VIII .1 VIII IOR

I

then either the reduction of the or anomer corresponding to formula VIII thus obtained under conditions such that N 3is reduced to NH 2without hydrogenolysis of the halogen atom at position 4 to give the o( or anomer corresponding to formula IX 1 2

R

this reduction being either followed by the reaction of the .L or k anomer corresponding to the formula IX with an halogeno ethyl isocyanate in order to convert the NH into NHCNHCH CH Hal and the action of a nitrite of an alkali H 0 0 metal, in particular sodium nitrite, in order to convert NH NHCH CHHa into NHF ir CH 2Hal in order to obtain 0

ONO

the compounds corresponding to formula I in which R represents a halogen atom and R represents O-C-R, 6 II 0 "'or followed by the hydrolysis of the O-C-R group present at

II

0 position 6 of the d. or anomer corresponding to formula IX and leading to the formation of an OH group, followed by the reaction of the c- or anomer corresponding to formula IX with an halogeno ethyl isocyanate in order to convert the NH 2 into NHCNHCHCH2 Hal' and by the action of 0 the nitrite of an alkali metal, in prt.icular sodiJm nitrite, in order to convert NHCNHC2 CH 2 Hal' into NHCNCH 2

CH

2 Hall in order g 0 to give rise to the compounds of the formula I in which R4 represents a halogen atom and

R

6 represents OH, or the reduction of the anomers VIII under conditions such that the halogen atom at position 4 is replaced by H and the N 3 group is 3 reduced to NH 2 to give rise to the or anomer corresponding to the formula X CH OR 0 0

H

2 H, OR 1 x

OH

J

16 this reduction being followed: Veither by the reaction of the t( or P anomer corresponding to formula X with an halogeno ethyl isocyanate in order to convert the NH 2 into NHCNHCH CH Hal' and the action of the nitrite of an alkali metal, 11 2 2 0 in particular sodium nitrite in order to convert NHCNHCH 2

CH

2 Hal' into NHCNCH 2

CH

2 Hal' 0

NO

in order to produce the compounds corresponding to formula I in which R, represents H, R represents O-C-R *or by the hydrolysis of the O-C-R group in posiion 6 of the 0 YC or anomer corresponding to formula X thus generation an OH group and leading to the formation of the t. or anomer corresponding to formula XI CH OH HH, OR 1

XI

OH

followed by the reaction of the o or F anomer corresponding to formula XI with an halogero ethyl isocyanate in order to convert the NH 2 into NHCNHCH2CH 2 Hal' and by the action of the nitrite of an alkali metal, in particular sodium nitrite, in order to convert NHCNHCH 2 CH2Hal' into NHCNCH2CH Hal II .I 0 ONO in order to produce the 1 or P anomer corresponding to formula I in which

R

4 represents H, and R 6 represents OH.

According to an attractive embodiment of the procedure of the invention for the preparation of derivatives corresponding to formula I, the reduction of the compounds corresponding to formula III such that N 3 is reduced to NH 2 and Hal is replaced by hydrogen is carried out by means of tributyl tin hydride in the presence of 2,2'-azo-bis-isobutyronitrile.

After the glycosylation of the compound corresponding to formula II the activated decivative of the acid RCOOH made t: react with the compound corresponding to formula II in order to introduce a benzoyl group at position 6 is advantageously benzoyl chloride,used in the presence of bis tributyl tin oxide.

Another preferred embodiment of the procedure of the invention for the preparation of nitrosourea derivatives .responding to formula I in which R 1

R

4

R

6 and Nu have the meanin !ndicated above comprises: the glycosylation of the compound corresponding to formula II in the form of the mixture of and B anomers by means of CH3OH in order to obtain the compound corresponding to formula XII

CH

2

OH

SoCH

XII

OH

S o the reaction of the compound corresponding to formula XII thus obtained with benzoyl chloride in the presence of tributyl tin oxide in order to produce the compound corresponding to formula XIII

CH

2 o 4 a O 0 OCH

XIII

N

3 3

OH

OH

the separation of the two and B anomers starting from the compound corresponding to formula XIII S the reaction of either the o( or A anomer with S0 2 C1 2 in order to obtain the or p anomer corresponding to formula XIV W OCH3 I

OH

and either the catalytYIc hydrogenation of the or anomer corresponding to formula XIV previously ex~plained in order to give rise to the 4Lor anomer corresponding to formula XV OH 2QC; I OM H, OCH 3 followed by -either the reaction of the tk or panomer corresponding to formula XV with anhalogeno ethyl isocyanate in order to convert the RH- 2 into

NH

2 en NHCNHCH 2

CH

2 Hal and by the action of the nitrite of an 2 1 0 alkali metal, in particular sodium nitrite, in order to convert NH NHCH 2

CH

2 Hal into NHCNCH CH Hal'

ONO

to give the V, o r Panomer corresponding to formula XVI 2 H, OCH 3 S. or by the reaction of the o( or anomer corresponding to formula XV with a base, in particular an alkali alcoholate in order to hydrolyse the benzoyl group and generate the d or B anomer corresponding to formula XVII

CH,OH

Cl CI 0

XVII

NH H, OCH

OH

followed by the reaction of the e or B anomer corresponding to formula XVII with an halogeno ethyl isocyanate in order to convert the NI12 into NHCNHCH 2 C H2 Hal' and by the action of the nitrite of an alkali 0 metal, in particular sodium nitrite, in order to convert NHCNHCH 2

CH

2 Hal' into 'NHCNC2 CH 2 Hal' I O1o to give rise to the or p anomer corresponding to the formula XVIII C1

XVIII

HNu OCH 3

OH

or the reduction of the d, or anomer corre3ponding to formula XIV by means of tributyl tin hydride in the presence of 2,2'-azo bis-isobutyronitrile to give the i or anomer corresponding to the formula XIX cH O 2 0 0

XIX

NH

2 i, OCH3

OH

ri--wla4-m.*-ir~ul~i3~;ES111 followed either by the reaction of the e or P anomer corresponding to formula XIX with an halogeno ethyl isocyanate in order to convert the NH 2 into NHCNHCH2 CH Hal and by the action of the nitrite of an alkali metal, O in particular sodium nitrite, in order to convert NHCNHCH CH Hal' into NHNCH 2

CH

2 Hal' 0 ONO to give the 0( or B anomer corresponding to formula XX ,OCH3 or by the reaction of the alpha or beta anomer corresponding to formula XIX with a base, in particular an alkali alocoholate, in order to hydrolyse the benzoyl group and generate the K or P anomers corresponding to formula XXI H, OCH3 followed by the reaction of the or P anomer corresponding to formula XXI with an halogeno ethyl isocyanate in order to convert NH 2 into NHCNHCH2CH2Hal' and by the action of the nitrite of an alkali O metal, in particular sodium nitrite, in order to convert NHCNHCH 2 CH Hal' into NHCNCH2CH 2 Hal' 0 ONO to give the o or B anomer corresponding to formula XXII 4- I 21

H

2 0H 0 Nu H, OCH3 XXII

OH

In carrying out the procedure according to the invention, the compounds corresponding to formula III

CH

2 -g R 0 Hal 0III N H, OR 1

OH

in which: R represents an alkyl group of 1 to 12 carbon atoms, or an aralkyl group of 7 to 12, and preferably 7 to 9, carbon atoms, possibly substituted on the aromatic nucleus by 1 or more, in particular, up to 3, halogen atoms, by 1 to 3 NO 2 or CF 3 groups or alkoxy groups of 1 to 4 carbon atoms, R represents an alkyl group of 1 to 6 carbon atoms, an aryl group unsubstituted or substituted on the aromatic nucleus by 1 or more atoms, in particular up to 3 halogen atoms, NO 2 ,oups, CF 3 groups or alkoxy groups of 1 to 4 carbon atoms, Hal represents a halogen atom, in particular chlorine, are new, and exist in the form of either their0( or P anomers.

Among the compounds corresponding to formula III, a preferreo group of compounds is constituted by those corresponding to formula II! in which: S R represents an alkyl group of 1 to 12 carbon atoms, R represents A paricularly useful new compound corresponds to the following formula i i I i .i 1 CH2q 0- H,

OCH

3 IIIa in the form of its d, or B anomer.

The compounds corresponding to formula III, in particular the two compounds corresponding to formula IIIa V, anomer or p anomer), arl: key intermediatesof the procedure and represent another feature of the present invention, In addition to the provisions which have been described, the invention also comprises others which will become evident in the subsequent description.

The invention relates not only to the new nitrosourea derivatives corresponding ~o the provisions already described and the procedure for their preparation but also to the new nitrosuurea derivates as therapeutically active substances as well as the compositions, in particular the therapeutic compositions, of which the said derivatives form part, The invention will be easier to understand with the aid of the supplement to the description which follows and which refers to examples demonstrating the procedure which is the subject of the present invention, However, it must be clearly understood that these examples of the implementation of the procedure are given only to illustrate the subject of the invention and they in t way constitute a limitation upon it.

EXAMPLE 1; Preparation of methyl 3-/(3-2-chloroethyl) 3-nitroso ureido 3,4-dideoxy- S-D-xylo-hexopyranoside (CI. 1675).

1) Preparation of methyl 3-azido 6-benzoyl 3-deoxy-p -D-gluco-hexopyranoside 29 g of 3-azido 3-deoxy D-gluco-hexopyranoside are dissolved in 600 ml of 1 N hydrogen chloride in methanol, the solution is heated under reflux for 2 hours and then evaporated to dryness in a vacuum.

The product is taken up in dichloromethane and washed with water until chloride ions have been completely removed. The organic phase is dried over sodium sulfate then evaporated to dryness in a vacuum to give a residue ~I L o f 31 g of glyco~ylated product which is used as such in the next step.

The crude product is dissolved in 250 ml, of anhydrous toluene. 90 ml of bis tributyl tin oxide are then added and the reaction mixture is then refluxed for 3 hours. The reaction mixture is cooled to -150' C and a solution of 40 ml of benzoyl chloride in 200 ml of methylene chloride is added dropwise, After being stirred for 214 hours, the reaction mixture is evaporated to dryness in a vacuum, then chromatographed on silica (eluant: CH 2 C1 2 "CH 301,98*:2), This leads to the isolation, in the form of resinous substances, of, 7,5 g of methyl 3-azidQ 6-'bonzoyl 3-deoxy-P -D-gluco-hexopyranoside.

0 3,5 C,0,83 CHQl 3 7.0 g of methyl 3-azido 6-benzoyl 3-deoxy-iC-D-gluco-hexopyranoside.

-D 102,00 (c,1,14 CH 2 Cl.

2 I2) Lrejaration of meT!hvkl3-azIdo 6-benzoyl1 4 chlqro 3,4 1dideox g h1a to -h xvra os id e To 4,8 g (0,015 mole) of methy! 3-azido 6-benzoyl 3-deoxy- P D gluICO-hexopyranosiee in 250 ml of anydrous pyridine 12,5 ml (0.15 mole) of sulfuryl chloride are added dropwise at 00 C, The solution is st~irred for 18 hours at 00 C, then allowed to warm to rc'orr. temperatuz'e 2 C The reaction mixture is then poured onto 500 g of ice and extracted with dichloromethane, The organic phas' is washed with 1 N sulfuric acid, then~ with water, dried over sodium sullate, filtered and evaporated, The oily residue Is purified by chromatography on silica using the eluant hexane-ethyl acetate 4:1 to give 4.05 g of white crystals, C 14

H

16 C1N 3 0 5 i 341,7 mp! 84-86' (hexanq-ethyl acetate) D+ 2.i30 (c,0.88 CHud 3 3) e m etitnhok -amino 6-benzoyl3- d~ex-P-D-xvlQhexopyranoside To 4.05 g (0.012 mole) of the preceding compound in 200 ml of anhydrous toluene is added a zo-bis -isobu t~f rolt rile (700 mg, 4,27 mmoles), followed, under nitrogen and dropwise, 1 q tributyl tin hydride (12.6 ml, 47 mmoles).

The reaction mixture is r,4fluxed for 2 hours and then evaporated uinder r~educed pressure, 24 F -After chr-omatogra'phy on silica using the eluant dichloromethaneamnioniacal methanol 19:1, 3 g white crystals are isolated, C 14H1 NO 5:281.3 mp; 1120-1170 -13, 60 (c,O0,3% CU 3

OH)

4) Preparation of m~thyl 3-am ino 3,4 dideoxy- -D-xylo-hexopyranoside To 3 g (0,0107 mole) of the preceding compound in methanol (45 ml) is added a molar solution of sodium methylate (5 ml), After being stirred for 2 hours at room temperature the reaction mixture is evaporated under reduced pressure, Ihe residue is purified by chromatography on silica using the eluant dichloromethane-amnioniacal methanol 17;3.

g of cr'stalline product are isolalted.

CU7H1 N 177.2 mpt 155 0-1580 (acetonitrile) 43.50 (c,0,74 CH 3

OH)

Preparation of methyl 3-/3-'(2-chloro etyl 3ni'oso ureido! 3,4-dideoxy- A -D.yohxopyyno e x 10 -3mole of methyl 3-amino 3,4-dideoxy- P -D-xylo-hexopyranoside are dissolved in 2 ml of anhydrous DUF and 5 x 10- mole of 21-chioroethyl isocyanate are added dropwise at 00 C with stirring, After being stirred for 5 hours, the reaction~ mixture is evaporated to dryness in a vacuum, A residue is dissolved in 8 ml of formic acid. Q006 mole of sodium nitrite are added In small portions and with stirring to the solution maintained at 00 C, After 30 minutes 10 ml of water are Added and stirring is continued for 1 hour. The reaction mixture is poured Into 100 ml of pure ethyl acetate, dried over sodium sulfate and evaporated to dryness,4 The residue is purified by crystallization from isopropyl ether.

Analysis.* C H 1 C1N 0 311.7 Yield,. 490/0 O 5.70 CH 3 010 EXAM1PL.E 2t Preparation of mathyl 3-.r-(Z-chloro ethyl)-3 ottroso ureido7 6-bernzoyl 3,4-dideoy' p -D-xylo-hexopyranoside (Cl, 1674)w Methyl 5-4mino 6-benzoyl 3i4-dideoXy- P -P-.Xylo-he-xopyranos1Je is prepared as described in example 1, atop 3, them the amino obtained 1.s treated as described in ex~ample 1, step Analysis: C 7H 22GiN 30 7: 415.83 Yield: 6 0 mp: 108 0 e7- 7,3 (c,0.28 CH OH) U 3 1EXAMPLE 3: Preparation of methyl 4-chioro 3-/"3-Q(-chloroethyl) 3-nitrosoureido7 3,4-dideoxy- P -D-galacto-hexopyranoside (CI: 1803), 11) orpratnf methyl 3-aminc 6-benzovl A-chioro 3,4-dideo~y~~ alacto-hexopyranos i de To 3 g of methyl 3-azido 6-benzoyl 4-chioro 3,4 dideoxy- -D-galaatohe-Xcpyranoside in 100 ml of absolute ethanol and 2 ml of triethylamine 500 mg of 10% palladiu'm on charcoal are added under nitrogen and the react4-n mixture is stirred in an atmosphere of hydrogen for 24 hours.

After the catalyst has been removed the filtrate is evaporated; the residkie is puriid by chromatography on silica.

The product ts recrystallized from 'Athyl e~her, Analysis: 14H1 ClNO 5:315.5 Yield: 82% 0 0 mp: 160 -1620 a: 11,0 (C,0.85 CHClI) 2) Preparation of methyl 3-atino 4- chloro 3,4dideo- -D ga Aedoprano side The debenzoylatiort is cat ried out~ according to the standard procedure using sodium methylate The product obtained ha the following propertiest Analysis: C 7

H

14 CIN0 4 ,211.5 mpt 1580 .1620 -4.0 (c,0.84 CHCl 3 3) Preparation o fMe~y _,chloro__3-/3'-_(2-chloro ethyl) 3.rnitroslo uraidodlad(to-hexopyranoside The preparation was oarried out as ;,as described in example 1, step startitng from the compound obtained in step 2 above, An~alysis: C 10

H

17 C1 2 N 3 0 6 t 346.17 mpt 14$0-1t480 LCl 30 (c,0.65 WeH), EXAMPLE 4: Preparation of methyl 3-~(2-chloro ekhyl) 3-nitroqu-ureidn7 3,4dideoxy- OC -D-xylo-hexopyranoside 1677) 1 Preparation of methyl 3-azido 6-benzoyl 3-o(-D-gluco-hexop ranoside 29 g of 3-azido 3-6eoxy D gluco-hexopyranoside are dissolved in 600 ml of 1 N methanolic hydrogerhloride and the solution is heated at reflux for 2 hours, then evaporated to dryness in a vacuum.

The product is taken .ip in dichloromethane and the solution is washed with water until chloride ion can no longer be detected. The, organic phase is dried over sodium sulfate and evaporated to dryness in a vacuum to give a residue of 31 g of glycosylated product used as such in the Inext step.

The crude product is dissolved in 250 ml of anhydrous tojuene, 90 ml of bis tributyl tin oxide are then added and rhe mixture is refiuxed for 3 hours, The reaction is cooled to 150 C and a solution of 40 ml of benzoyl chloride in 200 mil, of dichloromethane are added dtopwise. After being stirred for 24 hours, the reaction mixture is evaporated to dryness In a vacuum L id the residue obtained is chromatographed on silica (eluan( CH 2 Ct 2 "CH 3 0H/9$: 2) This results in the isol,-tion, In the form of vesinous substances, -o&o ehl -aio 6bool -ex' Dglchxprnsd 7.0 g of methyl 3-azido 6-berizoyl 3-deoxy -P-glucohexopyranoside.

2) Preparation of methyl 3-a zido 6-benzoy~l 4'-choro~~ i -ddeoxy-d-Dgalacto-hexo py ranoside.

To 4.8 g (0.015 mole) of methyl 3- azldo 6-bqnzy 3-deoxy-o -Dgluco-hexopyranoside in 250 ml of anhydrous pyridine are added dropwise at 00 0 12,5 ml (0.15 mole) of sulfuryl chloride. The solution is stirred for I8 hours at 00 G, then allowad to warm to room temperature.

The reaction mixture is then poured onto 500 g of ice and extracted with dichloromethane.

The organit. phase is washed with 1 N sulb'uric acid, them with water, dried, over sodium sulfate, filtered and evaporated, The oily residue is purified by chromatography on silica using the eluant hexane-ethyl acetate 4:1 to give 4.05 S of white crystals,

C

14 16

CN

3 O mp: 104-1060 (hexane-ethyl acetate) 140,5' (c,1.3 CHCl 3 27 3) Preparation Of Methyl 3-amino 6-benzoyl 3,4-diden.::y- ot- ,,ylohexopy ranos ide To 4.05 g (0.012 mole) of the preceding compound in 200 ml of anhydrous toluene is added azo-bis -isobu ty ronit rile (700 mg, 4.27 minoles) followed, dropwise and under nitrogen, by tributyl tin hydride (12.6 ml, 47 mmoles) The reaction mixture is heated at reflux for 2 hours and then evaporated under reduced pressure.

After chromatography on silica using the eluant dichloromethaneaminoniacal methanol 19:1, 3 g of white crystals are isolated.

4) Preparation of methyl 3-amino 3,4-dideoxy-t..-D-xylo-hexopyranoside To 3 g (0.0107 mole) of the preceding compound in methanol (45 ml) is added a molar solution of sodium methylate (5 After being stirred for 2 hours at room temperature the reaiction mixture is evaporated under reduced pressure. The residue '9 purified by chromatography on silica using the eluant dichloromothanie-ammoniacal methanol 17:t3.

g of crystalline product are isolated, MP 130 0-134 0 C 1630 (c'0'9 Gild 3 I 5) riepa':,aton of methyl 3-/3-(-chloroethyl) 3-nitroso ureido/ 3,4 d~deoxy-01.-D-xylo-hexopyranosid x 10- mole of -nethyl 3-amino 3,4-dideoxy- oi(-D-xylo-hex<opyranoside are dissolved in 2 ml of anhydrous DMF and 5 x 10- mole of 2-chloroethy, isocyamate are added dropwise at 00 C with stirring. After being stirred for 5 hours, the reaction mixture is evaporated to dryness in a vacuum.

residue is dissolved in 8 ml of formic acid. 0,036 mole of sodium nitrtte are added in small portions and with stirring to the solution maintained at 0 0 C. After 30 minutes 10 ml of watei are added and stirring is continued for 1 hour, The reaction mixttire is poured into 100 ml of pure ethyl acetate, dried over sodium sulfate nd e:vaporated to dryness.

The residue is purified by crystalliz.tion from IsopropNl ether.

Analysis: CIO H 18 C1N 3 0 6 311.7 mp: 1090 1100 +120.6' (c,0.48% MeOH).

EXAMPLE Preparation of methyl 4-chioro 3-/3--(2-chloro-ethyl) 3-nit'roso ureido7 3,4 dideoxy-o( -D-galacto-hexopyranoside (CI. 1676).

1) Preparation of methyl 3-amino 6-benzoyl 4-chioro 3,4-dideoxy-V -D- 2.alacto-hexopyranoside To 3 g of methyl 3-azido 6-benzoyl 4 -chloro 3,4 dideoxy- V, -0galacto-hexopyranoside in 100 ml of absolute ethanol and 2 ml of triethylamine 500 mg of 10% palladium on charcoal are added under nitrogen and the reaction mixture is stirred in an atmosphere of hydrogen for 24 hours.

After the catalyst has been removed the filtrate is evaporated; the residue is purified by chromatography on silica.

The product is recrystallized from ethyl ether.

Analysis: C 14H 18CNO 315.5 mp: 1610-1640 0 D+ 118.5 (c,1,25 CHCl 3 2) Preparation of methyl 3-amino 4-chloro 3,4 dideoxy- 6(-D-galacto- Ihexopyranoside The debeRoylation is carried out according to the standard procedure using sodium methylate.

The product obtained has the following properties: Analysis, C 7H 1ClNO4 211.5 mp:i 1350-1380 1840 (c,0.92 Me0H).

25I3) Peprtino mtyl4choo__________ 3 Prearaion f mehyl4-choro3-/3-(2-chloro-ethyl) 3-nitroso ureido! 3,4 dideoac- b( -D-galacto-hexopyranoside The preparation was carried out as described in example 1, step starting from the compound obtained in step 2 above, Analysis; C 10H 17Cl 2N 30 6 346.17 mp: 129'0_ 130' 139.30 0. 5 CH 3

OH).

PHARMACOLOGICAL STUDY; In order to test the antitumor activity of the substances of the invention described above use has primarily been made of murine leukemia 1,L1210. Use has also been made of the tumor melanoma B16, in p.grticular, the products of the invention have been tested ag, he melanoma B16 Iby administering them by the I.P. and by the I,V. routes.

La7 29 The products of the invention have also been tested against the colon carcinoma C38 by administering them by the I.P. and I.V. routes.

In practice, the biological effects of the novel nitrosourea derivatives corresponding to the present invention were tested as follows: PROTOCOL FOR TREATMENT WITH THE PRODUCTS OF THE INVENTION T. MATERIALS AND METHODS A. Tumors used Two murine tumors were used for the "in vivo" studies: the leukemia L1210 and the melanoma B16.

1) LEUKEMIA L1210 Animals The experiments were also performed on female mice which were specific pathogen-free of either the line DBA/2JIco or line B6 D2 Fl/JIco (first generati'n hybrids between the lines C57BL/6 and DBA/2).

The line used is specified for each series of experiments.

Tumoral graft On the day of the graft (by convention day 0 DO) an inoculum of 1 x 105 tumor cells in a volume of 0.2 ml is administered to each mouse by the intraperitoneal route This inoculum is prepared by diluting ascites fluid taken from the peritoneum of a donor female mouse in NCTC 109 medium (Eurobio Laboratories Paris, France), counting of the cells in a MALASSEZ cell under a microscope 5 and adjustment of the concentration to 5 x 10 cells per ml.

4- Distribution in experimental groups After the tumor graft has been made, the mice are distributed at random in cages of 5 animals. Subsequently, these cages containing 5 mice are themselves distributed at random into a control group (control) and groups treated with the products of the invention.

2) MELANOMA B16 Animals The experiments were alwafs carried out on SPF female mice B6 D2 Fl/JIco (first generation hybrids between the lines C5BL/6 and DBA/2).

Tumoral graft i i i On the day of the graft (Day 0) an inoculum of 2 x 10 tumor cells in a volume of 0.5 ml is administered to each mouse by the intraperitoneal route This inoculum is prepared from a sub-cutaneous tumor exjcsed from a donor female mouse. After excision the tumor is fragmented by means of a pair of scissors in the NCTC 109 medium. After filtration through sterile gauze in order to remove large cellular fragments, the homogeneous cell suspension obtained is counted by means of a MALASSEZ cell and diluted to the desired concentration (4 x 106 tumor cells per ml) by dilution with the NCTC 109 medium.

Distribution in experimental groups This was performed in the manner described previously for the leukemia L1210.

B. Protocol for treatment The doses of the products of the invention used in the different experiments are expressed in milligrams per kilogram of body weight.

The products to be injected were dissolved in isotonic sodium chloride solution, Two protocols for treatment were used in the different experiments: either a single injection on Dl, by the intraperitoneal or intravenous route, or 3 I.P. injections on DI, D5, D9.

In each experiment the animals of the control group received 1 or more injections, depending on the experimental protocol used, by the same route or of the same volume (0.2 ml/ 2 0 g) of the vehicle not containing the active principle (isotonic sodium chloride solution).

II. EXPRESSION OF THE RESULTS For each experiment a table specifies: the number and percentage of the total of the mice surviving to the T/C x 100 for the treated groups.

T representing the mean survival time of the mice in the treated group C representing the mean survival time of the mice in the control group (control).

III. COMMENT The strains of mice used, the experimental protocols and the 8 r mode of expression of the results are in accordance with directive 271 F of the Division of Cancer Treatment" (November 1983).

IV. PHARMACOLOGICAL RESULTS 1) Toxicological study: The results relating to toxicology are brought together in Table I below: TABLE I CI LD LD 50

LD

90 mg/kg mg/kg mg/kg 1674 )40 approximately.

1675 20 1676 )20 1677 2) Activity of the products CI. 1674, 1675, 1677 by the the leukemia L1210 USA strain and the melanoma 816, I.P. route on Substance of the invention CI, 1674: L1210 USA strain B 6

D

2

S

I

I,P? treatment on Dl, DS, D9 3 x 1.25 mg/kg, 3 x 5 mg/kg T/C Number of survivors at 3 x 1.25 111 0/10 3 x 5 130 0/10 Substance of the invention CI. 1675: L1210 USA strain B6D2F I.P. treatment on Dl, D5, D9 3 x 1,25 mg/kg, 3 x 5 mg/kg T/C Number of survivors at 3 x 1.25 164 0/10 3 x 5 >600 5/10 Melanoma B16 B6D F I.P. treatment on Dl, D5, D9 3 x 10 mg/kg T/C Number of survivors at 28 8/10 Substance of the invention CI. 1676: L1210 USA strain B6D 2F 1 62 1 3 x 1.25 treatment on Dl, D5, D9 3 x 1.25 mg/kg, 3 x 5 mg/kg Number of survivors at 0/10 0/10 I.P. treatment on D1, D5, D9 3 x 10 mg/kg Number of survivors at 3/10 Melanoma B16 B D2F 1

T/C

202 Substance of the invention CI, 1677: L1210 USA strain B6D F1- 621 3 x 1.25

T/C

157 600 I.P. treatment on Dl, D5, D9 3 x 1.25 mg/kg, 3 x 5 mg/kg Number of survivors at 0/10 6/10 I.P. treatment on DI, D5, D9 3 x 10 mg/kg Number of survivors at 7/10 Melanoma B16 B 6 2

F

1

T/C

257 3) Activity of the substances of the invention toward melanoma B16 after administration by the intravenous route: The activity of the substances of the invention was also tested by administering the said substances of the invention by the I.V. route.

ml of an homogenate of the tumor (Ig/10 ml) is administered to 33 B6C3F1 female mice by the subcutaneous route) on day 0. The products of the invention and the substances used for comparison are administered on days 3, 7 and 11 in isotonic solution.

In Table II below the results are presented relating to the activity of the substance CI. 1675 administered by the I.V. route on the growth of the melanoma B16 implanted subcutaneously, in comparison with

R

carmustine BCNU marketed under the name of BiCNU.

In Table II below and in the subsequent tables, T represents the mean weight (or median) of the tumors in the treated mice, C represents the mean weight (or median) of the tumors in the control, untreated mice and the ratio T/C% corresponds to T/C x 100.

TABLE II Product Dose Median weight of Median weight of T/C mg/kg/in, the t the or o day cn day 3 (mg cn day IC1675 20 36,0 688 8,8 23,0 1 044 13,4 32,0 2 025 25,9 BCNU 20 40 2 581 33,0 Controls 0 40 7 812 100 In this case it should be noted that the smaller the ratio the better is the activity, in the ideal case, the weight of the tumor in the treated mouse is 0 mg and hence the ratio T/C% is equal to 0.

The results obtained show that the substances of the invention are active when they are administered by the I,V. route, a finding which makes them quite suitable for use in human therapy, 4) Activity of the substance of the invention CI. 1675 administered by the I.P. route to mice bearing the colon carcinoma C38, implanted subcutaneously.

34 A tumor fragment is implanted on day 0 in female B D2F mice 1 9-22g).

The treatment with all of the products is administered by the I.P. route on days 2 and 9 which constitutes a delayed treatment. The weight of the tumors is calculated on day Th-e re.ults are oresented in Table III below.

j TABLE III Product Dose Median weight T/C Numbe. of mice without mg/kg/inj, of the tumor on t-.mor/total day 20 remarks (mg) IC 1675 30 0 0 7/7 and 5 dead mice 0 0 7/9 272 67 1/10 TCNU 30 10 premature dead 0 0 7/10 7,5 266 65 2/10 Controls 0 405 100 0/32 Table III above illustrates the considerable efficacity of the substance of the invention CI. 1675 against the colon carcinoma C38 when it is administered by the I,P. route, This activity is predicative of an activity in man for the treatment of various tumors.

Antitumor activity of the substance CI.1675 against the colon carcinoma C38: comparison of the intravenous and intraperitoneal routes, With a view to verifying the antitumor activity of the product of the invention CI.1671 when it i? administered by the I.V. route, another experiment was performed in which this product was administered by the I.P, route and the I.V. route for the purposes of comparison, This experiment is carried out under particularly difficult conditions, the first day of the treatment corresponding to the eighth day after the implantation of the carcinoma C38, whereas normally the treatment starts on the day of implantation of the tumor (DO) or 1 day after (Dl) the implantation of the tumor.

The median weight of the tumors on the first day of treatment (day 8) is about 12 mg (tumors varying from 8 to 40 mg). The animals are treated on days 8, 12 and 16 depending on the different routes indicated.

Ihe results relating to the action of the substance of the invention CI. 1675 in mice bearing the adenocarcinoma 38 of the colon are brought together in Table IV below.

TABLE IV I.P, Route I.V. Route Ds edian wt. Mice tMidjan wt. Mice SProduct Dos of the ithout of the T/C% without mg/kg/inj, tumor I ttor/total turor Jtumor/total J 120 J27 J20 J21 J20 J27 J20 J27 120 127 J20 427 S S1C 1675 20 13 172 2 1 1/8 2/8 0 0 0 C 6/8 6/6 88 726 15 5( 0 0 0 0 0 7/8 7/7 324 1099 57 84 0 0 23 352 4 2 0/8 0/8 SBCNU 20 196 274 34 21 0/8 0/6 OWOLS 0 586 1295 100 0/24 586 1295 100 0/24 It can be seen that not only is the activity of the substance of the invention towards the carcinoma C38 remarkable but that it is even higher after administration by the I.V, i ute than after I,P. administration, since at doses not producing toxicity (15 mg/kg/injection),a total regression of the tumor can also be observed. In fact, on the day a measurable tumor can no longer be observed at the beginning of the Ctreatment and on the 27th day there was no sign of recurrence, This is demonstrated by the number of mice without a tumor on the 27th day.

GENERAL CONCLUSION The experiments in animals performed with the substances according to the invention give very interesting results in the models used tumor L1210 USA and melanoma B16, and excellent results in the model of the colon carcinoma C38.

All of the results obtained 'ith these models show that the substances ,~e 36 of the invention have a wider spectrum of activity than the other antitumor substances known at present, Furthermore, the .compounds of the invention offer the remarkable advantage of bringing about the regression of animal tumors very difficult to treat with the known antitumor agents presently available.

The compounds corresponding to formula I above are hence therapeutically active substances and, from this point of view, they represent another aspect of the present invention.

The compounds according to the invention are particularly suited to the treatment of various human cancers, particularly those which respond to chemotherapy. They are particularly suited to the treatment of broncho-pulmonary tumors, tumors of the ENT sphere, digestive tumors (gastric, pancreatic, colic and rectal), breast tumors, genital tumors, bone tumors (osteosarcomas, reticulo-sarcomas), melanomas,hemato-sarcomas (Hodgkin and non-Hodgkin lymphomas), multiple myelomas.

The invention also relates to the pha-maeutical composition containing the new substances mentioned above in combination with a pharmaceutical vehicle appropriate to the chosen mode of administration, The invention 2C relates particularly to injectible sterile solutions suitable for administration by injections or intravenous perfusions. The invention relates in particular to physiologicall acceptable aqueous-alcoholic solutions.

The products according to the invention may, for example, be made available in the form of a iyophilized powder, the administration of which requires that it be made up into a solution immediately before u., with the aid of a sterile alcoholic solvent. The solution thus obtained is then diluted with pyrogen-free sterile water ard then, before being administered by intravenous perfusion, the solution is diluted again with a solution of glucose or isotonic sodium chloride.

The doses administered must be sufficiently high so as to provoke an activity in at least a relatively large proportion of the patients suffering from one or other of the various forms of cancer which are or will become accessible to the chemotherapy without, however, exceeding those doses at which the substances would be too toxic.

1 L -II As an example, the doses administered systemically, in particular by perfusion, and expressed as mg/kg, may vary from about 0.1 to about 5 mg/kg, for example about 1 mg/kg.

The invention also relates to other forms of administration, in particular by the oral route (liquid or solid compositions).

These dose ranges obviously have only indicative value, It is of course to be understood that in this type of therapy the doses administered must be assessed by the clinician in each case, on which occasion the condition of the patient and his personal reactivity towards medicines is taken into consideration.

An example of the pharmaceutical preparation of the products according to the invention contains from 10 to 250 mg, in particular 50 mg, of at least one of the products accordig to the invention, presented in the form of a sterile lyophilized powder in combination with an ampoula of a physiologically acceptable solvent, in particular of physiological serum, containing 5 ml of the solvent per ampoula.

As a resilt of their particularly high activity, the subst-nces of the invention are also useful as reference compounds in pharmacological studies, in particular in those designed to compare the antitumor properties of substances under study in comparison vith a reference compound i i-i~ lr ~i~ r ii

Claims (14)

1. Nitrosourea derivativet z-crresponding to formula I CH 2 R 6 R, 0 Nu H, OR 1 I OH in the form of one or other of the two anomers, t. or R in which R I represents an alkyl group of I to 12 carbon atoms, or an aralkyl group of 7 to 12, and preferably :f 7 to 9,carbon atoms, possibly substituted on the aromatic nucle.is by 1 or more, in particular up to 3, halogen atoms, by 1 to 3 NO 2 or CF 3 groups or alkoxy groups of I to 4 carbon atoms, R 4 represents H or Hal, Hal being a halogen atom, in particular Cl, R6 represents OH or Q-C-R R representing an alkyl group of 1 to 6 carbon atoms, an aryl group unsubstituted or substituted on the aromatic nucleus by 1 or more, in particular up to 3, halogen atoms, NO 2 or CF 3 groups or alkoxy groups of 1 to 4 carbon atoms, Nu represents the group NH-C'-CHC2-CH 2 -Ha1 Hal' 0 NO S 2 representing a halogen atom identical with, or diffr'nt from Hal and being in particular, chlorine,

2. Nitrosourea derivatives according to claim 1, corresponding to formula I, in which R 1 represents an alkyj group of 1 to 12 caTbon atoms, represents a halogen atom, R 6 has the meanings indicated in claim 1.

3. Nitrosourea derivatives according to elaim 1, corresponding to formula I in which R1 represents a C0 3 group, U~--IYU 39 R represents a halogen atom, R 6 has the meanings indicated in claim 1.

4. Nitrosoueaa !erivatives according to clai 1, corresponding to formula I in which ~1 o represents a CH 3 group, represents Cl, has the meanings indicated in claim 1, Nitrosourea derivatives according to claim 1, corresponding to formula I in which represents a CH3 group, represents a halogen atom, represents OCR R having the meaning indicated in claim 1. II 0 R 1 R 4 R 6

6. Nitrosourea derivatives according to claim 1, corresponding to formula I, in which R represents a CH 3 group, R represents a hydrogen atom, R6 represents OH. 7, Nitrosourea derivatives according to claim 1, corresponding to formula I, in which R1 repr, nts a CH3 group, R represents a hydrogen atom, R 6 represents O-C-R R having the meaning indicated in claim 1

8. Nitrosourea derivative chosen from among those having the following formulae n i i 40 i! L

9. Process for the preparation of nitrosourea derivatives corresponding to formula I H 2 A 2 6 R. 0 H, OR 1 O 0 in the form of one or other of the two anomers, or k in which R represents an alkyl group of 1 to 12 atoms, or an aralkyl group 1 of 7 to 12, and preferably from 7 t.o 9, carbon atoms, possibly substituted on the aromatic nucleus by one or more, in particular up to three, halogen atoras, by 1 to 3 NO 2 or CF 3 groups or alkoxy groups of 1 to 4 carbon atoms, R represents Hal, Hal being a halogen atom, and in particular Cl, R represents OH or 6 0 R representing an alkyl group of 1 :o 6 carbon atoms, an aryl group unsubstitutkd or substituted on the aromatic nucleus by 1 or several, and in particular up to 3, halogen atoms, by 1 to 3 NO2 or CF 3 groups or alkoxy groups of 1 to 4 carbon atoms, Nu represents the group NH-C-N-CH 2 -CH 2 -Ha Hal' NO being identical with,or different from Hal and representing a halogen atom, in particular chlorine, characterized in that a) in a first series of reactions, the starting compound corresponding to formula II 0 CH2OtH 3 OH OH 08t ~i-i is made to react in the form of the mixture of the o( and anomers with a glycosylating agent constituted by an alcch)1 R OH, R 1 having the meaning indicated above, in order to convert the OH group in position 1 into the OR 1 group, the product thus obtained is treated with an activated derivative of the acid R-COOH in order to convert the OH group in position 6 iito the O-C-R group, Sthe and B anomers are separated by a standard method, each of the anomers, and B is made to react with a halogenating agent in order to introduce a halogen atom in position 4, the desired compound which is obtained being in the form of one of the anomers, or P and corresponding to formula III, 3CH Hal 0 N H, OR 1 III OH in which R and R have the meanings indicated in for-uila I, Ha' represents a halogen atom, b) subsequently, in a second step, the reduction of the compound corresponding to formula III is carried out in order to reduce N 3 to NH 2 this reduction being carried out, in addition,under conditions such that the halogen atom in position 4 is or is not reduced to hydrogen, this reduction being possibly followed by the removal of the O-C-R group in position 6 and its replacement by an OH group, 0 the compound obtained at the end of this second step existing in the form of one of the anomers, b or and corresponding to the following formula V: 4 3 R I 2R.6 0 H, OR1 in which R 1 R 4 R 6 have the meanings indicated in formula 1, c) final~v, in a third series of reactions, the r or anomer corresponding to formula IV is allowed to react withan halogeno ethyl isocyanate in order to convert the NHi 2 into F4HCNHCH 2 CH 2 Hal' and the product thus obtained is treated with 0 the nitrite of an alkali metal, in particular sodium nitrite, in order to convert NHCNHCH CH 2 Hl into NHCNCH CQH 2 Hal' I I 2C P1111 0 ONO in order to obtain the compiund corresponding to formula 1, in the (orm, of one of the anorners, c or B

10. Process according to claim 9 for the preparation of nttrosourea derivatives corresponding to formula I r'haracteriz?d in that it comprises the reaction of the compound corresponding to formula II in the form of tb1e mixture of the o( and anoners H 2OH A OH with a glycosylating agent constituted by R 1 0H, R 1 having the meaning indicateed in claim 9, in order to give rise to the compound corresponding to formula V 0~ the introduction in position 6, R having in order to )Ai~ld the OH of an acyloxy group of the formula O-C-R 11 the meaning indicated in claim 9, 0 compround corresponding to formula VI, CH 2' the separation of the compound VI itito the two anomers K, and P C 2 0 R H, OR 1 the reaction of the tt or 5anomer corresponding to formula VII with a halogenating reagent to give the t 4 o r anomer corresponding to formula 111) it, OR 1 then Neither the reduction of the o( or anotner thus obtained corresponding to formula III under conditions such that N 3is reduced to Nil2 without hydrogenolysis of the halogen atom at position 4 to give the tiL or anomer corresponding to formula IX Hal 0 H OR Ix this reduction being IAXeither followed by the reaction of the or fanomer corresponding to formula IX with an, halogeno ethyl isocyanate in order to convert 2 into NCHH 2Ha'and by the action of the nitrite of an alkali metal, in vart Lcular sodium nitrite, in order to convert NHCflHCH 2 CH 2 'HIr 1 into NHCNCH HHl 1111 H 2 C 2 H 0 ONO in order to c.itain the e',ompokind$ corresponding to formula I in which Rrepresents a halogen a(.om and R 6 represents0-R 0 or followed by the removal from the or Panomer corre;sponding to, torm.ula IX of the O- group in position 6 and its V 2placemeot 0 by an OH group, followed by the reaction of the K or anomer, corresponding to formula IV withanhalogeno ethyl isocyanate in 3O order to convert NH 2 into NH5jNHCH 2 CH 2 Hal' and by the 0 action of the nitrite of an alkpli metal, in particular sodium A'itrite, in order to convert NHCNHCIL 2 CH2 Hal' into NHCNCH 2 CH 2 HalI 8 ONO to give the compounds corresponding to formula I in which R 4represents a halogen atom and Rrepresents OH, .r iLLi-. ii i ii ii ~__IWIIII IIIII~- U_ 'I 46 r N Sor the reduction of the respective anomers such that the halogen atom in position 4 is the N 3 group is rec iced to NH 2 to give the corresponding to formula X CH O X S0 \W H, OR 1 III under conditions replaced by H and and P anomers X this reduction being followed either by to formula X the NH 2 into the reaction of the t( or anomer corresponding with anhalogeno ethyl isocyanate in order to convert NHCNHCH 2 CH 2 Hal' and by the action of the i) 2 nitrite of an alkali metal, in particular sodium nitrite, in order to convert NHCNHCH CH Hal' into NHCNCH2CH 2 Hal' ONO in order to obtain the compounds corresponding to formula I in which R 4 represents H, R 6 represents O-C-R 0 .*or by the removal from the P. or B anomer corresponding to formula X of the O-C-R group in position 6 and its replacement II 0 O by an OH group to give the A or B anomer corresponding to formula XI i CR OH 0 Q2 H, OR X OH followed by the reaction of the 4 or anomer corresponding to formula X1 with an halogeno ethyl isocyanate in order to convert NH1 into NHCNHCH CHHl and by the action of the nitrite of an alkali metal, in particular sodium nitrite, in order to convert NHc NHCH 2 CH 2 Hal' into NHCNC H 2 CH 2 Hal' 0 O1NO in order to obtain the t4 or anomer corresponding to formula I in which R 4represents H, and R 6 represents OH.

11. Process according to claims 9 an( 10 for the preparation of nitrosourea derivatives corresponding to formula I, characterized in that the reduction of the compounds corresponding to fl r-mula III such that N 3 is reduced to Ni 2 and Hal is replaced by hydrogen is carried out by means of tributyl tin hydride in the presence of 2,2'-azo-bis-isobutyronitril4.

12. Process according to claims 9 and 10 for the preparation of nitrosourea de.'ivatives, corresponding to formula 1, characterizeu in that the compound correspcnding to formula II is subjected, after glycosylation, to reaction with bis tributyl tin oxide and benzoyl chloride in order to introduce,.the benzoyl group in position 6 and to give rise to the compound corresponding to formula III in which R represents a phenyl group in the form of the mixture of G~and anomers, which may subsequentliy be separated.

13. Process according to claim 9 for the preparation of nitrosourea derivatives corre~pondtng to formula I 4S CH 2 6 R 0 Nu H, OR 1 in which R 1 R 4 R 6 and Nu have the meanings indicated in claim 9. characterizpd in that it comprises tI glycosylation of the compound corresponding to formula II in the form of the mixture of its V, and anomers FH 2 Oil by means of CH OH in order to produce the compound corresponding to formnula XII OCH 3 the reaction of the compound c~orresponding to formula XII previ~ously obtilned with benzoyl chloride and tributyl tin oxide in order to obtain the compound corresponding to formula XIII 1 kL- CH 2 -D 2N OCH 3 XIII 23 the separation of the compound corresponding to formula XIII ito its two and Aanomers. the reaction of each of the V, and anomers with SO 2('1 in order to obtain the and anomers corresponding to formula XIV H, OCH 3 XIV anTd ,either the catalytic hydrogenation of the o4. and Fanomers corresponding to formulda XIV previously obtained to give the 4c and anomers corresponding to formula XV rlil 0xv HI OCH3 OH followed ,.either by the reac tion of the and P anomers corresponding to formula XV With an halogeno ethyl isocyanate in order to convert Nil 2 into NHCNHCH 2 C1 2 Hal' and by the action of the nitrite of 11 0 an alkali metal, in particular sodium nitrite, in order to convert o50 NHCNHCH 2 CH Hal' into NHCYCH 2 C82 Hal' to give 0 ONO the and P anomers corresponding to formula XVI CH2 0 1 0 XVI H, OCH3 o. Or the reaction of the t4 or anomers corresponding to formula XV with a base, in particilar an alkali alcoholate to remove the benzoyl group and generate the 4 or anomers corresponding to formula XVII CH OH 1 2 XVII H, OCH 3 followed by the reaction of the oc or anomers corresponding to formula XVII with an halogeno ethyl locyanate in order to convert N2 ino NHNHCHCH Hal and by the action of the nitrite of an O alkali metal, in particular sodium nitrite, in order to convert NH NHCH 2 CH Hal into NHCNH C Hal' o dA o oC or anOmers crresndinrg to forula XVIII C1 XVII Nu H 0C0 No :o give the I L L-L_ 5S1 'or the reduction of the V, or Panomers corresponding to Iformula XIV by means of cributyl tin hydride in the presence of 2,2'-azo- bis-isobutyronitrile to give the 4 or anomers corresponding to formula XIX Nh 2 K' 2 I 100 followed ,.either by the reaction of the bL or anomers corresponding to formula XIX with an haidgeno ethyl isocyantate in order to convert NH 2 into NHC1NHCH 2 CH 2 Hal. and by the action of the nitrite of an alkali g metal, in particular sodium nitrite, in order to convert rqHCNHCH 2 CH,, Hal' into NHCNCH 2 CH 2 Hal to give the tk and anomers corresponding to formula U 0 0 x H, OCH 3 or the reaction of the t4 or anomers corresponding to formula XIX with a base, in particular an alkali alcoholate, in order to remove the benzoyl gro~up and generate the t4 and P anomers corresponding to formnula XXI 52, 04:113 2:) follot,,d by the reaction of the V, and F anomers corresponding to formula XXI with an halogeno ethyl isocya1nate in order to convert the NH 2 into NH jHCH 2 CH 2 Hal and by the action of the nitrite 0 of an alkali metal, in particular sodium nitrite, in order to convert N~CH C~ la.' ino NHF NQH 2 H 2 Hal 0 ONO to give the VL and anomrnes corresponding to f~rmutla XXII CH2 0" ,On 1 3 XXII Compound$ corresponding to formula III Htl OR I OH in which I i -53 R represents an alkyl group of 1 to 12 carbon atoms, or an aralkyl group of 7 to 12, and preferably from 7 to 9, carbon atoms, possibly substituted on the aromatic nucleus by 1 or more, and in p..rticular up to 3, halogen atoms, by 1 to 3 NO 2 or CF 3 groups or alkoxy groups of 1 to 4 carbon atoms, R represents an alkyl yroup of 1 to 6 catbon atoms, an aryl group unsubstituted or substitutedon the aromatic nucleus by I or more, in particular up to 3, halogen atoms, by 1 to 3 NO 2 or CF 3 groups or alkoxy groups of 1 to 4 carbon atoms, Hal represents a halogen atom, and in particular chlorine, in the fomn of either its or anomer. Compounds according to claim 14 corresponding to formula III in which R1 represents an alkyl group of to 1. car';on atoms, R represents '16. Cimpound of the formula CH2O9 O, 0CH in the form of its t or p anomer.

17. The compounds according to any one of claims 1 to 8 as therapeutically active substances,

18. Pharmaceutical composition characterized in that comp'iges any one of the substances according to elaims 1 to 8, in comibination with a pharmaceutically acceptable vehicle.

19. Pharmaceutical composition according to clait 18, characterized in that in contains 10 to 250 mg, and in particular 50 mg, of at least one of the substances according to any one of ii 4, 54t claims 1 to 8, pres~nted in the form of a sterile lyophilized powder, in combination with an ampoula of a physiologically acceptable solve nt, in particulat physiological serum, -ontaining 5 ml of so, nt per ampoule. DATED this 21st day of April 1988 SANOF1 ENWD, WATERS SONS PATENT ATTORNEYS QUEEN STREET MELBOURNE, VIC. 3000. CertifIy fl-A t- 1; n~~!yfdp~L L tru~o thoA 1.