4-substituted 1,3-diglycidyl urazole compounds,their production and pharmaceutical preparations containing them

4-Substituted 1,2-diglycidyl urazole compounds, their production and pharmaceutical preparations containing them

HENKEL KGaA

C. 59174 and Israel Patent 59453

German Offenlegungsschrift No. 29 07 349/relate£ to cytostatically active pharmaceutical preparations containing as their pharmacologically active principle triglycidyl isocyanurate (TGI) and/or TGI derivatives in which the hydrogen atom of the carbon in the 2-position of the glycidyl group may be replaced by an alkyl radical Containing, from 1 to 4 carbon atoms. Compounds of this type are distinguished by the fact that the three N-atoms of the isocyanuric acid ring are substituted by epoxy10 group-containing glycidyl radicals which may even be substituted in the 2-position by an alkyl radical containing from 1 to 4 carbon atoms.

German Offenlegungsschrift No. 30 37 094.6 relates inter alia to cytostatically active pharmaceutical preparations containing compounds corresponding to the following.general formula

R

I °γ^Ν_χ/° glycidyl θ glycidyl in which R may have the following meaning: alkyl, aryl, aralkyl', alkaryl, cycloalkyl, which radicals may if desired even be heterocyclic, unsaturated and/or substituted 20 by .at least one of the following substituents: halogen, hydroxyl, amino, N-substituted amino, mercapto, alkyl

ו .

mercapto, aryl mercapjso, alkyl sulfoxy, aryl sulfoxy, alkoxy, aroxy, acyloxy and heterocyclic^ radicals. The glycidyl radical has the meaning defined above.

The present invention is based on the observation that compounds of analogous structure, but derived from urazole as the parent substance, also show surprisingly strong cytostatic activity which can even exceed that of triglycidyl isocyanurate.

In a first embodiment, therefore, the present invention relates to new N-substituted di- and triglycidyl urazole compounds corresponding to the following general formula

N— N

Ra Rb in which each of the radicals Ra and Rb represents a glycidyl radical of the formula

CH₂

CH₂ (II) and the radical R is a glycidyl radical of the formula (II) above or a radical Z which is a phenyl radical, a lower alkyl radical or a lower hydroxyalkyl radical further substituted with a hydroxy, lower alkanoyloxy or morpholino group.

In a first particularly preferred embodiment of the invention, the three radicals R present in N-substitution are a glycidyl radical corresponding to general formula II.

.1 ^{In the} production of 1,2,4-triglycidyl urazole (TGU), three diastereomers are theoretically formed because only two of the three glycidyl groups have the same chemical surroundings. ־

<img file="IL64844A_D0001.tif" />

<img file="IL64844A_D0002.tif" />

Statistically, the following isomer distribution should be* obtained without any mutual influence:

RRR’ SRR' RSR* RRS'’

SSS' RSS' . SRS'SSR’

<img file="IL64844A_D0003.tif" />

: . 2 :1 (0) (v) .(a)

These three isomers are present in the crude product obtained after the treatment with alkali. According to.the results of column chromatography as described in the following, the TGU isolated asprimary product from the column consists of only two diastereomers which are present in a ratio of approximately 1:1.׳

Identification: HPLC:silica gel/methylene chloride + 2.5 % of methanol.

The two diastereomers, which it is!intended to refer to as a- and 0-TGU, differ only slightly in their ,retention times. a-TGU has slightly shorter retention time than

0-TGU. In the inverse system (mobile phase:water + 1.32 % of THF), the ratios are reversed and separation becomes slightly better so that this sytem may also be used for preparative separation.

a- and 0-TGU also differ in their melting points and in their ¹H-NMR-spectra - particularly significantly in the range from 4.0 to 4.4 ppm.

The third diastereomer, which it is intended to refer to as γ-TGU, was eluted from the column after the a-/fl-TGU.

It may al&amp;o be isolated from the’mother liquor after recrystallisation of the crude product obtained by the process bed hereinafter by purification using column chromatography.

This γ-TGU is liquid at room temperature and is present in the crude product in a proportion of from about 15 to 20% (and not 50% as had been expected).

The physical data of the three diastereomers are set out in the following Table.

. Table

<td> 20</td><td> TGU\</td><td> M.p. or 20 . n_D^:</td><td colspan="5">¹H-NMR-double bands between 4 and 5 ppm (coupling const., Hz)</td>

<td></td><td> a-</td><td colspan="2"> 104 °C (extrapol.)</td><td> 4.17</td><td> (3.0);</td><td> 4.35</td><td> (3.0)</td>

<td></td><td> ־8</td><td> 115-116*C (</td><td> I ״</td><td> 4.07</td><td> (2.4);</td><td> 4.23</td><td> (2.4)</td>

<td></td><td> Ύ”</td><td> 1.5088,.</td><td></td><td> 4.55</td><td> (2.5);</td><td> 4.66</td><td> (2.5);</td>

<td> 25</td><td></td><td></td><td></td><td> 4.08</td><td> (6.3);</td><td> 4.18</td><td> (6.3);</td>

<td></td><td></td><td></td><td></td><td> or as</td><td> a double</td><td> doublet</td><td></td>

<td></td><td></td><td></td><td></td><td> I 4.12</td><td> (12.0);</td><td> 4.60</td><td> (12.0)</td>

The integral of this range corresponds to approximately 30 two. protons.

The present invention also relates to these three diastereomers of 1,2,4-triglycidyl. urazole, to the process for their production and to their use a^ described־ ־in the following. Mixtures of these diastereomers which differ in 35 their composition from the diastereomer mixture originally obtained by synthesis also fall within the scope of the present invention. In this connection, the present invention relates in particular to diastereomer mixtures of TGU which consist essentially of two of the ibovementioned diastereomer types. Thus, the scope of the invention includes in particular the mixture consisting essentially of a- and β-TGU obtained by column chromatography and its use.

In another important embodiment of the invention, only two glycidyl radicals corresponding to general formula II are present on the urazole ring, whilst the third radical is a radical Z having the meaning defined above.

According to the invention, the molecular weight of a radical Z of the type in question preferably does not exceed the limits indicated below. Thus, the molecular weight of this radical Z is best no more than about 750 and, more particularly, no more than about 500. It can also be of advantage for the molecular weight of the radical Z not to exceed a value of approximately 300 or even a value of approximately 200.

The present invention also relates to processes for producing the new N-substituted polyglycidyl compounds corresponding to general formula I and to pharmaceutical preparations characterised in particular by cytostatic activity and containing compounds corresponding to general formula I.

־ \

\ \ - 7 The mechanism by which the compounds used in accordance with the invention act has not been explained in detail. Presumably, the glycidyl groups which are present both here and in the triglycidyl isocyanurate according to DE-OS No. 29 07 349 are of crucial importance so far as the cytostatic effect is concerned. All the compounds of general formula I according to the invention are characterised by the presence of at least two such glycidyl groups. In addition, the widely variable radical R may optionally be present in the class of compounds in question. It is possible that the distribution of lipophilic and hydrophilic preferences is influenced through this radical Z and that the absorption of the compounds by the organism may thus be controlled to a certain extent. However, the new substituent Z introduced in accordance with the invention is not limited in its meaning to this possible effect.

According to the foregoing definition, the radical Z is a hydrocarbon radical which may also contain heteroatoms. The heteroatoms in question are, in particular, N or 0. This radical preferably contains in all no more than 15 carbon atoms, preferably no more than 12 carbon atoms and, more preferably, no more than 8 carbon atoms. Radicals containing tip to 6 or, preferably, even only up to 4 carbon atoms may be particularly interesting. These figures should be interpreted independently of the particular structure and merely apply to the sum of all the carbon atoms in the radical in question.

In one particularly preferred embodiment of the invention, the radical z represents an alkyl radical optionally substituted as above. This alkyl radical may be straight-chain or branched and, excluding its substituents, preferably contains no more than 8 carbon atoms. In this embodiment of the invention, particular preference is attached to those compounds of general formula I in which the radical Z represents unsubstituted alkyl containing from 1 to 6 carbon atoms or a corresponding alkyl radical substituted by hydroxyl, lower alkanoyloxy or morpholino.

Radicals substituted in this way may be substituted once or several times by the abovementioned groups. From 1 to 3 of the abovementioned substituents are preferably present on the particular radical Z. In one particularly preferred case, compounds of general formula I containing substituted alkyl radicals of the abovementioned type are used in the pharmaceutical preparations according to the invention.

One particularly preferred embodiment of the invention relates to compounds corresponding to general formula I in which one of the radicals R, i.e. the radical Z, represents a monosubstituted or disubstituted alkyl radical of the abovementioned type selected from the following group: monohydroxy alkyl, dihydroxy alkyl, and alkanoyloxy hydroxy alkyl. The alkyl radical may advantageously contain up to 7 carbon atoms, preferably from 3 to 7 carbon atoms and, more particularly, 3, 4 or 5 carbon atoms.

According to the invention, preferred compounds of

64B44/3 general formula I may be those In which the radical Z represents Btraight-chain or branched unsubstituted alkyl containing up to 6 carbon atoms and preferably up to 4 carbon atoms. The radicals in question are, in particular, methyl, ethyl, propyl, isopropyl and the corresponding Cj-radicals. In addition, compounds of general formula I in which one of the radicals R is a monosubstituted or disubstituted alkyl radical of the type mentioned above containing in particular 3 carbon atoms and preferably contains at least one hydroxyl group, are particularly preferred. In these compounds, therefore, at least one hydroxy group is always present, preferably adjacent another substituent on the radical in question, whilst the other two radicals R represent the glycidyl radical corresponding to general formula II. In another preferred embodiment of the invention, these substituting groups in the radical Z are distributed between the 2-position and the 3-position of the radical R in question. In this connection, the hydroxy group may be either in the 2-position or in the 3-position. Other particularly preferred, correspondingly substituted compounds of formula I contain no other substituting group apart from the hydroxyl group in the radical Z or contain as further substituents hydroxyl or an alkanoyloxy radical.

If, in addition to the hydroxyl group, Z contains an alkanoyloxy radical, this radical also preferably contains up to at most 10 carbon atoms, the preferred limit in this case, too, lying at 8 carbon atoms. It is particularly preferred to introduce no more than 5 carbon atoms into the molecule at this point. Alkyl radicals containing the corresponding number of carbon atoms are also preferred in the case of alkanoyloxy radicals. The alkanoyloxy radicals are preferably derived from monocarboxylic acids having the abovementioned number of carbon atoms and the abovementioned structure.

The medicaments according to the invention may with advantage contain individual, defined compounds corresponding \ to general formula I, although it has been found that activesubstance mixtures of several of the compounds falling within the scope of general formula I are also highly effective cytostatic agents. According to the invention, it can also be of advantage to use certain individual compounds or a mixture of several compounds corresponding to formula I in admixture with the TGI compounds disclosed in DE-OS No.

29 07 349 and in German Patent Application P 30 37 094.6.

The production -of the compounds according to the invention may he carried out in several ways and is another subject of the present invention:

1. Introduction of the glycidyl groups of general formula

II in N-substitution into the ur.azole ring. To this end, urazole, i.e. the parent compound of formula I containing an -NH-group instead of each of the N-glycidyl groups, is first produced in known manner. The hydrogen on the 10 nitrogen atom is then replaced by the glycidyl group.

There are above all two fundamental possibilities for carrying out.this final step of the reaction. One of these is directly to introduce the glycidyl group by reacting the NH-group with epihalohydrins, particularly 15 epichlorohydrin or epibromohydrin, followed by dehydrohalogenation. The other possible method completes the formation of the molecule in the two reaction steps.

Firstly, the‘corresponding ally!-substituted intermediate products are formed, after which the allyl group is epoxidised 20 in a concluding step.

The reaction of -NH-groups with epihalohydrins is widely reported in the literature. The reaction may be carried out in the presence of a small quantity of a quaternary ammonium compound as catalyst (cf. for example 25 Houben-Weyl Methoden der Organischen Chemie,Vol. 14/2 (1963), 497, 547). Particularly suitable quaternary ammonium compounds belong to the class of phase transfer catalysts. The compounds in question are known to be quaternary ammonium compounds of pronounced lipophilic 30 character attributable in particular to the presence of sufficiently large organic residues in the quaternary ammonium compound. Detailed information on phase transfer catalysts 4 may be found, for example in Phase Transfer Catalysis in Organic Synthesis by W.P. Weber and G.J. Gobel, Springer 35 Verlag, Berlin, Heidelberg, New York, 1977 and in

1 ׳ י י

I

Phase Transfer Catalysis by E.V. Dehmlow and S.S. Dehmlow, Verlag Chemie., Weinheim, Deerfield Beech (Florida), Basel,1980.

The phase transfer catalysts are preferably used in quantities of from about 0.1 to 10% by weight, more preferably in quantities of from 0.5 to 5% by weight and most preferably in quantities of. from 0.5 to 3% by weight, based on the urazole compound, in the following reaction step, dehydrohalogenation - which to some extent is also brought about by excess epihalohydrin - is completed by the addition of bases, preferably alkali hydroxides.

In the second process, the urazole compound is not directly reacted with the epoxide compound. Instead, it is first reacted with allyl halides which, although corresponding to the glycidyl radical in the compounds of 15 general formula I, contain an olefinic double bond instead of the epoxide group, after which the allyl-substituted ׳urazoles formed are epoxidised. Epoxidation may be _tcarried out in known manner using peracids. A related reaction is, for example, the reaction of cyanuric acid 20 with allyl halides as described in US Patent No. 3,376, 301. The epoxidation of allyl isocyanurates using peracids is described for example in Houben-Weyl loc. cit. Vol. 6/3, pages 385 et seq. It may be carried out for example in the presence of a small quantity of a quaternary ammonium compound 25 as catalyst.

. . The reaction of the urazole or of the mohosubstituted .¢ Tin» 1 iiwmt 11>1> <11 1111 ummil^tii <1Γ tin» nuiiiti 11111)911 luled ' ‘J י' י- ' . . ' , urazole compound (cf in this connection Section 3. below) with epihalohydrins or with allyl halides is best carried out at temperatures in the range from about 50 to 150°C and 30 preferably at temperatures in the range from about 70 to about 125 °C.

The reaction may be carried out in an excess of the epihalohydrin compound as solvent or in ^olar aprotic solvents which partly dissolve at least. one of the reactants 35 and which are not reactive to the reactants. Particularly

J —1 wn II !JBUIIBI I ill .J. ^,־TJ ־־־.־', JT. --JILJI JILl. II L,l 11 1|Ι1|]ΙΙ.Ιΰ appropriate solvents are any of the dialkyl formamides, particularly the lower dialkyl formamides, such as dimethyl formamide. The preferred reaction time is from 1'to 10 hours and, more particularly, from 2 to 5 hours.

The complete dehydrohalogenation of the halohydrins formed as intermediates may be obtained by the addition of solid, powdered alkali, preferably NaOH, or by the addition of highly concentrated aqueous solutions. This dehydrohalogenation step is carried out either in excess epihalohydrin or after the removal thereof by distillation under reduced pressure in a polar aprotic solvent, such as for example dimethoxy ethane, diglyms or dimethyl formamide, at temperatures in the range from -10 to 60°C and preferably at temperatures in the range from 0 to 45°C.

. _י Epoxidation of the allyl groups using peracids is also preferably carried out in solvents. Solvents suitable for this purpose are, once again, polar solvents, for example halogenated hydrocarbons or alcohols.

Suitable reaction temperatures are normally in the range from 0 to 50°C and, more particularly, in the range from about 10 to 3p°C. The peracid is best used in a substantially equivalent quantity or in only a slight excess, n-chloroperbenzoic acid is readily available as a commercial product and.suitable for carrying out the reaction. The reaction time is generally of the order of 24 hours or longer, for example up to 48 hours.

If unsubstituted urazole is used as starting material in these reactions, it is possible to obtain the triglycidyl־־ substituted urazoles.

2. There are various possibilities for producing urazole derivatives according to the invention in which two of the radicals R in general formula I represent a glycidyl radical and the third radical R represents the radical Z. . One possibility is to react triglycidyl uralole (TGU) with a substoichiometric quantity of water, alcohol, primary and/or &lt;.11! nimni ¹μ*» w . secondary amines,. mercaptans, imines, imides, carboxylic acids, hydrogen halide and the like or hydrogen.

In view of the similarity of the three glycidyl groups in the TGU, this reaction always leads in the first instahce 5 to product mixtures which in turn may show therapeutic activity. : However, it is also possible and part of the process according to the invention as described in the following to separate the corresponding compounds of general formula iTrom these mixtures by suitable separation techniques, for example by preparative thin-layer chromatography or column chromatography.

In the course of these reactions, a glycidyl group is converted into the radical Z of the compounds corresponding to general formula I. A monohydroxyalky.l radical Z is 15 ־formed in the reductive treatment of the glycidyl group with hydrogen or with hydrogen donors. suitable hydrogen donors are, for example, hydride compounds, for example complex borohydrides, such as sodium borohydride. In the other cases mentioned, the triglycidyl starting compound is 20 reacted with a substoichiometric quantity of a nucleophilic compound H⁺A , resulting in the formation of a disubstituted radical Z which, in addition to a hydroxyl group, contains the radical A as a second substituent, normally on the atom adjacent the hydroxylated C-atom of the radical R.

The reaction of the glycidyl groups of a structurally similar compound, namely triglycidyl isocyanurate(TGI), with nucleophilic reactants of the type in question is state of-the art and is described, for example, in Angew.Chemie 80, 851 (1968). In the prior art, however, 30 this reaction is specifically carried out on more than only one epoxide group of the TGI and is used for example in the crosslinking of epoxide resin systems on an industrial scale. By contrast, the process according to the invention is preferably carried out under conditions which enable the yield to be increased as far as possible towards 1:1 reaction products nnd which provide for the nubnoquont isolation nnd recovery of I.Ihmjo 1:1 reaction product» by nepa 171 tiny off unreacted parts of the starting material and more . advanced reaction products.which have formed through the . 5 reaction of more than only one epoxide group with the nucleophilic reactant.

In the reaction of TGU-compounds with nucleophilic reactants H⁺A of the type mentioned above, it may be difficult^to obtain the required 1:1 reaction products 10 in high yields because the three epoxide groups of the molecule of the starting compound are substantially equal to one another in their reactivity, with the result that in many cases the required diglycidyl compound is not formed as the main reaction product. Difficulties are also occasionally involved in efforts to increase the concentration of the required compound by reacting the .. triglycidyl urazole with a substoichiometric quantity of f nucleophilic־ reactant.

It has been found that the 1:1 reaction products can be produced surprisingly easily by reacting the triglycidyl urazole with an excess and preferably with a large excess of the nucleophilic reactant H⁺A , but prematurely terminating the reaction and separating off the excess of nucleophilic reactant, unreacted TGU and 25 co-formed diaddition and triaddition products. The ' crude diglycidyl product left behind may then be purified by conventional methods, for example by column chromatography. In this process, the nucleophilic reactant is preferably used in a 3- to 30-fold excess and, more particularly, in a 30 5- to 20-fold excess over and above the necessary quantity.

. The reaction may be carried out in solvents although, if desired, the excess of the nucleophilic׳ reactant may also serve as solvent. it the reaction is carried out in solvents, the solvents used are best substantially polar 35 solvents which, preferably, are not reactive under the conditions selected for the process. The solvent may also be immiscible with water. Particularly suitable solvents are, for example, halogenated hydrocarbons, particularly chlorinated hydrocarbons. . The reaction temperature is normally in the range from about 30 to 120*C and preferably in the range from 40 to 100°C and, in one particularly suitable embodiment, is selected in such a way that the epoxide content of the reaction mixture falls by half in 4 to 5 hours.

Both in this process and in the other processes described in the following, the purification of the 1:1 reaction product containing two epoxide groups and its recovery from the mixture of reactants is generally an essential step of the process according to the invention.

3. An extremely elegant, general process for producing the compounds corresponding to general formula I is based on the reaction of the mono-N-substituted urazole compound with epihalohydrins. The production of mono-N-substituted urazole may be carried out by methods known from the literature. For relevant literature, see for example Org. Synthesis, Vol. 51, 121 (1971).

The substituent introduced into the urazole generally corresponds to the radical Z in the compounds of general formula 1. The two glycidyl groups are then introduced in a subsequent reaction. To this end, the monosubstituted urazole is reacted with the corresponding epihalohydrin compound, .for example with epichlorohydrin, followed by dehydrohalogenation, or with allyl halide, followed by epoxidation of the double bond, as described above, in I section 1. with reference to the triglydidyl urazole. In another embodiment, therefore, the present invention relates to a process for the production of Nsubstituted polyglycidyl urazole compounds corresponding to the following general formula

<img file="IL64844A_D0004.tif" />

in which R_a and R^ are as defined above, characterised in that the two glycidyl radicals corresponding to general formula II are introduced in N-substitution into urazole or into a urazole mono-N-substituted by the radical Z or a triglycidyl urazole containing glycidyl radicals corresponding to general formula II is subjected to a partial reaction with water or alcohols and the reaction products formed, which correspond to general formula I, are separated off from the reaction mixture and recovered as such.

If, in this process, the glycidyl radicals corresponding to general formula II are introduced into urazole or into mono-N-substituted urazole, this may be done by directly reacting the urazole compound optionally substituted by the radical Z with epihalohydrins, these epihalohydrin compounds corresponding to the glycidyl radicals of general formula II, followed by dehydrohalogenation, or alternatively by initially reacting the, urazole compounds with corresponding allyl halides and subsequently converting the allyl radicals or ο

the allyl radicals substituted by Rj into the glycidyl group by epoxidation, preferably using peracids. All the observations made in the foregoing apply in the same way to the characteristics of the compounds corresponding, to general formula I, the radicals R and Z and glycidyl and the reactants involved in their formation.

The compounds corresponding to general formula I, particularly in purified form and in bulk, are new compounds. They are suitable for use in medicaments. However, the new polyglycidyl urazole compounds may also be used in other fields where polyglycidyl-substituted compounds normally occupy a position of importance. Fields of this type are known to include in particular the field of plastics and, more particularly, the field of epoxide resins.

If the polyglycidyl compounds contain a radical Z in addition to two glycidyl radicals of general formula II, this radical Z may be in the 1-, 2- or 4-position. The 4-position may be particularly important for these compounds containing the radical Z, above all on preparative grounds.

The compounds corresponding to general formula I are suitable for the treatment of malignant neoplasms. The compounds may be administered in individual doses of from 1 to 200 irg/kg.

Certain individual compounds corresponding to general formula 1 or mixtures thereof may be used. Their use in admixture with other active components, for example TGI, also falls within the scope of the present invention.

The compounds of general formula I used in accordance with the invention occur in various stereoisomeric forms. In principle, any ol these various forms are suitable for the purposes of the invention. In this connection, they may be used either in admixture or even in the form of certain isolated isomers. ¹

For use as cytostatic agents, the active substances should be applied by moans of suitable vehicles. Suitable vehicles are the auxiliaries and excipients normally used for pharmacological preparations. In the present case, suitable vehicles are aqueous systems, optionally in conjunction with compatible.glycol ethers, such as glycol.

monoethyl ether or butylene glycol methyl ether or propylene glycol methyl ether, particularly if the active principle is to be administered parenterally. In the case of oral administration, the usual pharmaceutical auxiliaries and vehicles may be used, providing they are sufficiently compatible with the glycidyl compounds.

In animal experiments, good results were obtained with freshly prepared aqueous solutions administered i.p. or i.v.

The compounds used in accordance with the invention are effective against various forms of leukaemia and malignant neoplasms, such as carcinoma of the lung, carcinoma of the colon, melanomas, ependymoblastoma and sarcomas. In .some cases, the new urazoles were found to be distinctly superior to cyclophosphamide and fluoruracil.

Combination therapy in conjunction with other cytostatic 20 . agents, such as derivatives of nitrogen mustard .gas or even fluoruracil,' is possible.

It may be said quite generally of the compounds of general formula I containing a radical Z used in accordance with the invention that this radical Z shows or should show 25 little or no reactivity with the epoxide groups of the .glycidyl substituent(s) on the ring system of general formula I, at least under normal conditions or at least with cooling.

.This ensures that the active components used in accordance .30 with the invention are sufficiently stable in storage and do not undergo any undesirable reaction culminating in destruction of.the epoxide groups. This rule should also be observed in particular in the selection of any substituents present on the radical R. |

The following are examples of the radical Z in the ־ ' i

I • , —».ויין 'V! PT' W¹ 5-. _י V'^WJII UIIIIIWWJW

- 21 cytostatically active compounds of general formula 1 used in accordance with the invention: methyl, ethyl, propyl, butyl, pentyl, hexyl, the corresponding isomeric radicals, such as isopropyl, isobutyl, tert.-butyl and isopentyl.

In the context of the invention, suitable reactants for converting a glycidyl group in the tlriglycidyl urazole to form a substituted radical Z are, quite generally, alkanols, such as methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, sec.-butanol, tert.-butanol, 1-pentanol, 2-pentanol, 3-pentanol, 2-methyl-l-butanol, 3-methyl-l-butanol, 2-ethyl2-butanol, 3-methyl-2-butanol, 2,2-dimethyl-l-propanol,

1- hexanol, 2-ethyl-l-butanol, 4-methyl-1-pentanol, 4-methyl-

2- pentanol, 2-methyl-l-pentanol. Examples of polyhydric alcohols are ethylene glycol, 1,2-propane diol, 1,3-propane diol, 1,4butane diol, 1,2-butane diol, 2,3-butane diol, 1,3-butane diol, 2-butene-l,4-diol, 2-butyne-l,4-diol, 1,5-pentane diol, 2-methy11,4־-butane diol, 2,2-dimethyl-l,3-propane diol, hexane diol, 2,5-dimethyl-3-hexyne-2,5-diol, glycerol, 1,2,4-butane triol, 2-hydroxymethyl-2-ethyl propane diol, 2-methyl-2-hydroxymethyl-l,3-propane diol, pentaerythritol.

An example of the reaction of the glycidyl group with an amine is morpholine.

The polyglycidyl-substituted urazoles according to the present invention are normally present in the medicament mixtures according to the invention in concentrations of up to about 20% by weight, based on the medicament mixture. The concentration range from 0.05 to 10% by weight for example .1 is suitable, the range from 0.05 to 5% by weight being particularly suitable.

The percentages quoted in the following Examples are percentages by weight, unless otherwise indicated.

⁵ EXAMPLES

EXAMPLE 1

15.2 g (0.15 mole) of urazole, 0.4 g of tetramethyl ammonium bromide and 0.5 g of Benzalkon A (a mixture of alkyl benzyl dimethyl ammonium chlorides) were boiled under 10 reflux for 3 hours in 416 g (4.5 moles) of epichlorohydrin.

After cooling, 28.8 g (0.72 mole) of powdered sodium hydroxide were added to the solution, followed by stirring for 6 hours at 45°C. The solution was then filtered off under suction from the deposit and evaporated to dryness 15 under reduced pressure at 40°C, after which the residue was dissolved in a little methylene chloride and subjected to column chromatography (eluent - methylene chloride:methanol 95:5) on silica gel (Merck). *

The individual fractions were combined after TC-analysis. 20 After concentration by evaporation, the second collective fraction (substance with the second highest R -values) Γ yielded 1.5 g of. 1,2,4-triglycidyl urazole which, after recrystallisation from ethyl acetate, still melted at 93 to 94°C.

Epoxide number observed: 17.1 calculated: 17.8

The structure is confirmed by elemental analysis and mass spectrum. EXAMPLE 2

1.53 g (0.01 mole) of 4-phenyl urazole (prepared in accordance with Org. Syntheses, Vol. 51, page 121 (1971)), 0.08 g of Benzalkon A, 0.08 g of tetraethyl ammonium bromide and 50 g of epichloroHydrin are stirred for 4.5 hours at 60°C. After cooling to room temperature, followed by the addition of 20 g of molecular sieve 4 2, 0.88 g.(0.024

I « ιτΰΰ *f ¾¾-4 *־V™ jmillJ ,¾¾ l»RJi4AW I . 23 * mole) of NnOH in 1 ml of 11^0 Ln added dropwise, after which the reaction mixture is stirred for 1.5 hours at 45°C, filtered under suction and concentrated in vacuo at 40°C. Yield: 2.75 g . % EpO: 8.9

The crude reaction product is separated by column chromatography.

Column height: 40 cm, diameter; 4 cm Filling: silica gel 60 (Merck)

Eluent: m&amp;thylene chloride:ethylacetate:methanol 3:2:1. The zone having ah R -value of 0.65 is isolated.

Γ

1.85 g of 1,2-diglycidyl-4-phenylurazole are obtained. % EpO: 11.9 (theoretical 12.1)

The structure is confirmed by the mass, IR- and

NMR-spectra.

.EXAMPLE 3

The following tests were carried out in accordance with the procedures laid down by the National Cancer i Institute,. Bethesda, Maryland 200014, as published in

Cancer Chemotherapy Reports Part 3, September 1972, Vol. 3, No. 2. The glycidyl compounds according to Examples 1 and 2 were used as active substances. The substance was freshly prepared in the form of an aqueous 1% injection solution immediately before application.

Tumor type P 388 (leukaemia) was induced i.p. in mice (10⁶ cells/mouse) in accordance with procedure 1200 (page 91c). The average period of survival of the untreated animals is. determined.

In further groups of tests, the active substance 30 is administered to correspondingly pretreated animals. The life of the treated test animals is significantly prolonged by comparison: with the average.period of survival . of the animals'which have not been treated with the. active *

substance. The prolongation factor T/C |n dependence upon 35 the dosage of the active substance is shown in the following

<td> 1</td><td> Table: Table Example Dose administered T/C-value (mg/kg)</td>

<td> 5</td><td> 1 50 298 25 &gt; 260</td>

<td></td><td> 12.5 230 6.25 180</td>

<td> 10</td><td> 2 200 200 100 150 50 120 25 120</td>

<td> 15</td><td> EXAMPLE. ,4</td>

<td></td><td> f2/3-dlhydroxypropyl) - . N-dihydroKyprop-y-lf-N* ,N-diglycidyl urazole</td>

<td></td><td> 5 g of triglycidyl.urazole (0.019 mole) are stirred</td>

<td></td><td> for 3 hours at 70°C in 50 ml of water. The solution is</td>

<td> 20</td><td> concentrated in a rotary evaporator and dried in a high vacuum. The colourless, oily crude product (5.8 g) is purified by column chromatography.</td>

Column: 35 x 5 cm

<td> 25</td><td> Filling: silica gel 60; 0.063 - 0.2 mm(Merck) Eluent: ethyl acetate 2 parts methylene chloride 2 parts methanol 1 part The desired compound is obtained in the form of a colourless oil. Rf: 0.43 in the above eluent on silica gel</td>

<td> 30</td><td> plates Yield: 30% by weight, based on crude product</td>

<td></td><td> % epoxide oxygen: 11.02 (calculated 11.14) IR,.MS and ^H-NMR confirm the structure.</td>

<td> 35</td><td> EXAMPLE 5 1 _י yl ' N-(2-hydroxy-3-propio1^bxy propyIT-N^r,N’^r-diglycidyl urazole</td>

5 g of triglycidyl urazole (0.019 mole) and 14 g of propionic acid (0*19 mole) are stirred for 3 hours at. 100 - 110°C in 100 ml of absolute toluene to which 5 g of molecular sieve 4 8 have been added. After filtration of the molecular sieve, the solution is concentrated. The residue is dissolved in methylene chloride and extracted by shaking twice with 50 ml of 10% soda solution. The methylene chloride phase dried over sodium sulfate is concentrated. The colourless oily residue obtained (5.2 gj is purified by column chromatography.

Column: 35 x 5 cm

Filling: silica gel 60 (Merck) 0.063-0.2 nun

Solvent: methylene chloride + 5% methanol

The desired compound is isolated in the form of 15 a colourless oil.

Yield: 21% by weight , Rf: 0.3 in the above-mentioned eluent on . silica gel plates %epoxide oxygen: 9.:5 (calculated 9.3)

IR and ^H-NMR confirm the structure. EXAMPLE

N-(2־-hydroxy-3-morpholin-N-yl-propyl)-N*,N-diglycidyl urazole 5 g of triglycidyl urazole (0.019 mole) and 2 ml of morpholine (0.022 mole) are stirred for 3 hours at 50°C in

100 ml of absolute isopropanol. The solution is concentrated • and dried in a high vacuum. The yellowish, oily crude product (7.4 g) is purified by column chromatography. Column: 35 x 5 cm

Filling: silica gel 60 (Merck) 0.063 - 0.2 mm . 30 Eluent: . . methylene chloride + 20% of methanol

The desired compound is isolated in the form of a colourless oil. ז

Yield: ¹ 51% by weight .

Rf: 0.72 in the abole-mentioned eluent on silica gel plates % epoxide oxygen:

IR and /H-NMR confirm

Elemental analysis:

Observed Calculated

9.0 (calculated 9.0) the structure.

<td> 5</td><td> C:</td><td> 50.7</td><td> %</td><td> 50.55</td><td> %</td>

<td></td><td> H:</td><td> 6.2</td><td> %</td><td> 6.79</td><td> %</td>

<td></td><td> N;</td><td> 15.5</td><td> %</td><td> 15.72</td><td> %</td>

EXAMPLE

1.2- diglycidyl-4-methyl-urazole

8.5 g (0.074 mole) of 4-methyl urazole (prepared in accordance with R.C. Cookson, S.S. Gupte et al., Org. Synth. 51 (1971), page 121) and 2% of tetraethyl ammonium bromide are stirred for 4 hours at 80°C in 230 ml of epichlorohydrin (2.95 moles). After the addition of 65 g pf molecular sieve 4 8, the cooled solution is stirred for hours at 40-50°C with 24 g of 50% sodium hydroxide (0.3 mole). After filtration under suction, the filtrat^ is . concentrated‘by evaporation and the pale yellow, solid residue recrystallised from methanol. The 4.5 g (27% of 20 the theoretical) of white crystals obtained, which have an epoxide oxygen content of 14% (calculated 14.1%), melt at 90°C. IR, Ir-NME and MS confirm the structure.

EXAMPLE

1.2- diglycidyl-4-butyl urazole

The procedure is as in Example 7, except that 4-butyl urazole is used as the starting material. The crude . product is purified by column chromatography.

Column: . . 40 x 5 cm

Filling: silica gel 60 (Merck) 0.063 - 0.2 mm

Eluent: methylene chloride + 5% of methanol

36% by weight of pure product are isolated.

Rfi 0.65 in the above-mentioned eluent on silica gel plates.

% epoxide oxygen: 11.7 (calculated. 11.9)

M.p. of the white crystals: 49-52°C

<img file="IL64844A_D0005.tif" />

IR and ^H-NMR confirm the structure.

EXAMPLE 9

The three stereoisomeric forms of triglycidyl urazole (α-, ¢- and γ-risomer) are prepared and isolated as follows:

A mixture of 12.5 g (0.124 mole) of urazole, 174 g (1.88 mole) of epichlorohydrin and 0.25 g of tetraethyl ammonium bromide was stirred for 14 hours at 70 °C, the volatile constituents were distilled off under reduced ׳ pressure at a bath temperature of 40°C, the residue was taken up in 300 ml of methylene chloride, and the solution filtered, followed by the addition over a period of 5 minutes with vigorous stirring at 20°C of 70 g of a 30% sodium hydroxide solution. Stirring is continued for 1 hour at 20°C, after which the methylene chloride phase is separated off and the aqueous phase extracted three times with 100 ml of methylene chloride. The methylene chloride phases collected were concentrated by evaporation under reduced pressure (ultimately at around 1 mbar) at a maximum bath temperature of 40°C. The yield amounted to 23^2 g (69% of the theoretical). TGU is present in the form of a mixture of the three diastereomers (partly crystalline product) which, according to quantitative TC-analysis, contained 73% of a-/8-TGU and approximately 15% of γ-TGU.

Recrystallisation from ethyl acetate gave a-/B-TGU melting at 92-97°C (sintering at 87°C) in a yield of approximately 40% (composition: a-/g-TGU = 50:50 (HPLC). Further recrystallisation produced a gradual increase in the melting point to near the melting point of pure β-TGU.

The first mother liquor contained all the γ-TGU and some of the a-/6-mixture. The γ-TGU was obtained in a yield of 10% from the mother liquor by column chromatography (silica gel/C^C^ :CH^OH = 98:2). γ-TGU is a colourless \ 20 liquid having ahrefractive index n^ of 1.5088.

The a-/8-mixture obtained from the first mother liquors contained distinct concentrations of a-TGU.

As described in Example 3 experiments were carried out in mice with different compounds and the following results were obtained:

<td> Compound</td><td> Dose Administered mg/kg</td><td> T/C Value</td>

<td> B-TGU*</td><td> 50</td><td> &gt;300</td>

<td> .γ-TGU _</td><td>¹⁰⁰</td><td> 293</td>

<td></td><td> 50 up</td><td> to 210</td>

<td> a-TGU</td><td> High activity at less</td><td> than 50mg/kg</td>

<td> Example 4</td><td> 100</td><td> 300 ד</td>

<td></td><td> 50</td><td> 278</td>

<td> Example 5</td><td> 100</td><td> 250</td>

<td></td><td> 50</td><td> 230</td>

<td> Example 6</td><td> 200</td><td> 230</td>

<td></td><td> 100</td><td> 160</td>

<td> Example 7 .</td><td> 100</td><td> &gt; 300 ’</td>

<td></td><td> 50</td><td> 210</td>

<td> Example 8</td><td> 200</td><td> 243</td>

<td></td><td> 100</td><td> 180</td>

<td></td><td> 50</td><td>¹⁵³</td>

* All mice were healed