CS200249B2 - Process for preparing new quinazolondiurethanes - Google Patents

Description

(54) Method for the production of new quinazolone diurethanes

The present invention relates to novel quinazolone di-urethanes, to a process for their preparation and to their use as cytostatics.

Numerous quinazolone derivatives are already known to have hypnotic, soothing properties or cause muscle relaxation (see Angew). Chemie 21 (1962), 855-861]. In contrast, there were no known derivatives of the class of these substances which are effective against malignant growth.

It has now been found that the novel quinazolone diurethanes of formula 1

(I) where

R is C 1 -C 2 alkyl having very potent cytostatic properties can be obtained according to the invention by providing 6-isocyanato-2- (3-isocyanatophenyl) -3-phenyl-4- (3H) -quinazolone of the formula XI (III)

O

is reacted with an alcohol of formula III

R = OH where

R is as defined above, optionally in the presence of inert solvents.

When 6-isocyanato-2- (3-isocyanatophenyl) -3-phenyl-4- is used as starting material. - (3H) -quinazolone and methanol, the reaction sequence can be illustrated by the following reaction scheme:

The diisocyanato derivative of the formula II used according to the invention can be prepared in a manner known per se from the diamine of the formula IV

O

for example by phosgenation of the hydrochlorides in chlorobenzene at 80 to 120 ° C.

The diamine of the formula IV can be obtained in a manner known per se by catalytic hydrogenation of the corresponding dinitro compound. This dinitro compound can be produced by the previously described process (German Published Applications Nos. 1,809,174 and DOS 1,809,175) by reacting the corresponding nitroanthranilic acid and the corresponding N-substituted nitrobenzimide chloride in a polar solvent such as acetone in the presence of an aliphatic tertiary amine such as triethylamine , at a temperature ranging from 0 to 50 ° C or by reacting the corresponding nitroanthranilic acid with nitrobenzoyl chloride to give the corresponding benzoxazinone and reacting the intermediate with the corresponding primary amine.

In both cases, a first intermediate part, which can be thermally in organic3 200249 Kem solvent, for example glycerol, at a temperature above 100 ° C or by using dehydrating agents for example Ρ ^ ₂ 0 ⁷ N-methylpyrrolidone, and cyclized to the corresponding dinitro compound.

The alcohols of formula (III) used as starting materials are known.

Preferably, an excess of the hydroxy compound of formula III is used, but the reaction may be carried out in the presence of inert diluents such as dioxane, aromatic hydrocarbons, dimethylformamide, dimethylacetamide or N-methylpyrrolidone.

In the reaction of the compounds of formulas II and III, the temperature may lie within a wide range. Generally, the temperature is in the range of about 10 ° C to about 120 ° C, preferably 25 ° C to 80 ° C.

In the process according to the invention, the hydroxy compounds of the formula III, when not used as a solvent, are preferably used in an amount of 1 to 10 moles, preferably in an amount of 2.2 to 6 moles per mole of the compound of the formula II.

When the free hydroxy compounds of formula III are reacted with the compounds of formula II, it is recommended to add a small amount of a basic catalyst such as triethylamine, pyridine, triethylenediamine or dimethylcyclohexylamine. The catalyst is used in an amount of 0.1 to about 5% by weight of the total reaction batch.

In the following example, parts by weight have parts by volume as kilograms to liters or grams to milliliters.

Example

From 2- (3'-isocyanatophenyl) -3-phenyl-6-isocyanato-4- (3H) -quinazolone and methanol.

parts by weight of 2- (3'-isocyanatophenyl) -3-phenyl-6-isocyanato-4- (3H) -quinazolone and 2 parts by volume of triethylamine are heated to boiling with 200 parts by volume of methanol for 30 minutes. The reaction mixture is stirred into water and the product is recrystallized from ethanol.

Yield of 2- (3 '- (methoxycarbonylamino) phenyl) -3-phenyl-6-methoxycarbonylamino-4- (3H) -quinazolone: 36 parts by weight (81% of theory). Mp 243-245 ° C.

Analysis:

calculated: 64.86% C; 4.5.4 H; 18.0% O; 12.61% N;

found: 64.7% CI 4.7 * Hj 18.2% O 12.8% N;

2- (3'-Isocyanatophenyl) -3-phenyl-6-isocyanato-4- (3H) -quinazolone is obtained from 2- (3'-aminophenyl) -3-phenyl-6-amino-4- (3H) - quinazolone as follows:

164 parts by weight of 2- (3'-aminophenyl) -3-phenyl-6-amino-4- (3H) -quinazolone are suspended in 1000 parts by volume of anhydrous chlorobenzene. Dry hydrogen chloride is introduced into the suspension until at least an amount of hydrogen chloride is absorbed which is sufficient to form the dihydrochloride. Phosgene is then introduced at 80 to 100 ° C until a clear solution is formed. The reaction deer is then stirred at 100 ° C for a further half hour, purged with nitrogen and the clear solution is concentrated under vacuum.

The white product thus obtained is pure 2-O'-isocyanotophenyl) -3-phenyl-6-isocyanato-4- (3H) -quinazolone according to infrared spectrum. Melting point: 153-155 ° C.

The compound of the formula is obtained in an analogous manner

Mp .: 204-206 ° C, yield: 86%.

As already mentioned, the novel compounds have good cytostatic activity, making them useful in medicine, especially in the suppression of lymphatic leukemia. The new compounds therefore represent an enrichment of the current technique.

The good efficacy of the compounds of the invention was tested in a model of transplanted L1 210 lymphatic leukemia in mice (Table 1), and the experiments were performed as follows:

Mice of 18-22 g (strain B6D2F1) were administered intraperitoneally with 2 x 10 < 6 >

Treatment was initiated 24 hours after leukemia cell transplantation and a 4X intraperitoneal dose was administered every subsequent day.

The experiment time was 2 to 3 weeks.

To evaluate the results of the experiment, the survival index was determined as follows:

If the survival time of the 50 control group is considered to be 100%, the survival time of the 50 treated group χ 100 can be determined according to the sample χ 100 survival index = ^{11 1 1 1} - * of animals.

Assessment:

Values <100% indicate a reduced survival of the treated group of animals, i.e., toxic effects of the preparation.

Values> 100% indicate an increase in survival time of 50, and tumor growth suppression can be compared according to the height of the index.

Table 1

Compound of Formula I

Leukemia LI 210 Optimal dose mg / kg body weight at 4 doses intraperitoneally

Survival index

R = CH3 100 1 047

Rg CgHg 350,913

Oytostatic quinoxaline derivatives are already known (U.S. Pat. Nos. 3,455,920, 3,463,778, I. Med. Chem. St. 10 (1967), pp. 334-366]. Compounds of the invention are progressing over these known compounds in that they have significantly lower cytotoxicity with approximately the same cytostatic effect. This is clearly apparent from comparative testing of the cytostatic and oytotoxic effects of the following two substances:

Compound I 2- (3 * -methoxycarbonylamino) phenyl] -6-methoxycarbonylamino-4- (3H) -quinazolone (a compound of the invention);

Substance II 2,3-dihydro-6-nitro-2-phenyl-4- (1H) -quinazolinone (substance known from Example 1 of U.S. Pat. No. 3,455,920).

Testing is performed both in vivo on Walker carcinoma 256 (Test 1) and in vitro on suspension cultures of mouse L fibroblasts (Earle) (Test 2).

Test conditions

G e s 1

Wistar rats weighing 140-160 g are inoculated intramuscularly with a Walker carcinoma tumor suspension to the thigh of the right hind leg 256. Treatment with compounds I and II is initiated 24 hours after tumor transplant 5 times after Animals are sacrificed 10 days after tumor transplantation and weight is recorded tumor, weight change, and food intake. The efficacy of compounds I and II is expressed by the value of the tumor mass index (TG index), which is calculated according to the formula average tumor weight of treated groups of animals TG index = average tumor weight of the control group of animals.

When the average tumor weight of the control group is considered equal to 1, the TG indices have decimal values ranging from 0.0 to 1.0.

The wording of efficacy at different TGs is according to normal international practice:

desired efficiency 1.0 to 0.8 low efficiency 0.7 to 0.6 significant efficiency 0.5 to 0.4 good efficiency 0.3 to 0.2 very good efficiency 0.1 to 0.0

Test 2. L-cell suspension cultures with an initial cell count of 2 x 10 6 cells / ml are treated with compounds I and II at 1000, 100, 10, 1 and 0.1 µg / µl respectively. After 24 hours of assay, the cell number index is calculated according to the formula average number of viable cells in treated cultures index of cell numbers = average number in control cultures.

Results

Test 1

In Vivo Assay for Walker Carcinoma 256. Tumor mass, body weight and food intake indexes were determined after dosing with the same effect (equivalent doses).

Substance II has been shown to be significantly more toxic than flight I. This is evident from higher percent animal losses, a large reduction in body weight and, finally, a significantly reduced food intake following administration of substance II. For the sake of clarity, the corresponding values for substances I and II are given in Table II.

Table 2

Daily dose mg / kg 5x p. Therapeutic effect Animal losses (%)

Change in body weight (g)

Daily food intake per animal (g)

0,6 +27

17.0

100 75, 50

0.3 0.6 0.3

12.5 0 37.5 +9 -1 -16

12.7 11.7 6

Test 2

In vitro assay on L-cells

Cell number indices after administration of the same concentrations of compounds I and II were determined.

The cell number index after administration of Compound I has the following values after 24 hours of testing:

Concentration (/ ig / ml)

0.01

0.1

1.0

100 1,000

Cell Number Index

0.9

0.8

0.9

0.7

0.7

0.4

The corresponding values for substance II are as follows

Concentration (Aig / ml)

0.01

0.1

1.0

100 ALIGN!

000

Cell Number Index

1,2

1,2

1.1

0.8

0.5

0.3

It follows that substance II has a significantly stronger cytotoxic effect against L-cells than substance I. This explains the higher toxicity of substance II observed in vivo.

In vitro experiments also show that Compound I has a mechanism of action consisting in specific retention of mitosis in metaphase, whereas Compound II acts on cells only through its non-specific cytotoxic effect. Due to the specific mechanism of action, compounds I can be used to synchronize the cell cycle of tumor cell populations. This is not possible for Substance II, so that the progress of Substance I over Substance II is also evident.

Also known are urethane derivatives having antileukemic activity (DE 1 102 721). However, the compounds according to the invention, in particular substance I, are better than the most active compounds of the cited file, i.e., compounds III and IV,

(III) (IV) both in terms of their anti-leukemic effect when administered orally and in terms of their mode of action.

Advantage of substance I over other substances of similar structure can be demonstrated on two criteria, namely on the criterion of better antileukemic effect after oral administration and on the other

200249 8 mechanism of action criterion. Indeed, it can be shown that the mechanism of action of blocking mitosis does not have a comparative substance.

The demonstration of the advantages of Compound I was carried out by means of the following comparative tests using compounds III and IV.

Comparative testing of the effect of these three compounds was performed in vivo on the leukemia of P 388 mice, an internationally used conventional test tumor. The results of the assays are shown in Table 3. In these assays, the percent increase in survival time of the leukemic mice treated with the individual preparations over the corresponding control groups served as an effect parameter. The percentages given are summarized in the table.

The data presented in Table 3 show that a significant increase in the survival time of the leukelic mice is achieved with Compound I compared to Comparative Compounds III and IV, which is significantly more significant for oral administration than for intraperitoneal administration.

In addition to these in vivo assays, the mechanism of action of these three compounds on Sarcoma MCS 4 and Sarcoma 180 cell cultures was also tested in vitro. The results are shown in Table 4 · *

From the data in this table, it is clear that of the three compounds tested, only Compound I has a mitosis blocking effect. This effect is still evident even at low concentrations of the preparation of 0.05 µg / ml. Both naphthoquinone compounds III and IV are cytostatically active only through their cytotoxic effect (the effect of deadly cell damage) and are therefore not suitable for the controlled control of tumor cell kinetics. In view of the clinical applicability of these three test compounds, only substance I meets the prerequisites for the controlled control of tumor cell kinetics.

Table 3

Comparison of the efficacy of Compound I compared to Compounds III and IV in suppressing leukemia P 388 in mice in vivo.

Parameter: Percentage prolongation of survival (PPDP) of leukemic mice compared to control groups.

Tabulated values: Optimum efficacy over the entire dose range used in the assay.

Dose range mg / kg Method of administration Optimal PPDP (%) Optimal PPDP (%) Optimal PPDP (#) AND III IV 50 to 500 after. 104 96 78 2.5 to 250 i.p. 26 7 15 Dec 2.5 to 250 i.p · 30 30 22nd 1 to 500 after. 104 59 59

2002 49

Table 4,,

Testing the mechanism of action (M) of compounds I, III and IV on cell cultures of Sarcoma 1CS4 (1) and Sarcoma 180 (2)

Concentration of preparation in nutrient medium (Aig / ml); 1 1 000 to 1

10 to 0.01

Preparation

Mechanism of action of Sarcoma MCS 4 ED ^ q WM • on cells ⁽¹⁾ (> ug / ml)

Mechanism of action of Sarkom 180 ED ^ q WM on cells ⁽²⁾ (ug / ml)

Even blocking mitosis undetectable

III fatal damage

Fatal damage

The novel compounds are preferably administered orally, mitosis blocking 0.05 lethal injury 5 lethal injury 5, possibly also intraperitoneally.

The novel compounds can be used either as such or in combination with non-toxic inert pharmaceutically acceptable carriers. Suitable dosage forms for the administration of the active compounds in combination with various inert carriers are tablets, dragees, capsules, granules, aqueous suspensions and emulsions, non-aqueous emulsions and suspensions, syrups and the like. These carriers include solid diluents or fillers, aqueous media and the like. various non-toxic organic diluents and the like. Of course, sweeteners and the like may be added to tablets and other dosage forms. Therapeutically active compound should be in the above-mentioned forms present in a concentration from approximately 0.1 to 99, preferably from 0.5 to 90% by weight, based on _(the total mixture.

The dosage forms are manufactured in a manner known per se, for example by mixing the active ingredients with diluents and / or carriers, optionally using emulsifiers and / or dispersants.

Carriers and / or auxiliary agents include, for example, the following substances:

Water, nontoxic organic solvents, or diluents, such as paraffins (e.g., petroleum fraction), vegetable oils (e.g., underground oil and sesame oil), alcohols (e.g., ethyl alcohol, glycerin), glycols (e.g., propylene glycol, polyethylene glycol); solid carriers such as natural mineral meals (e.g. highly disperse silicic acid; silicates), sugar (e.g. raw, lactic and grape sugar), emulsifiers such as non-ionic and anionic emulsifiers (e.g. polyoxyethylated fatty acid esters, polyoxyethylated fatty alcohol ethers, alkylsulfonates and arylsulfonates), dispersants (e.g. lignin, methylcellulose, starches and polyvinylpyrrolidone), glidants (e.g. magnesium stearate, talc, stearic acid and sodium lauryl sulfate).

The tablets may of course contain, as already mentioned, additives in addition to said carriers; such as sodium citrate, calcium carbonate and middle calcium phosphate, together with various other ingredients such as starches, preferably potato starch, gelatin and the like.

Lubricants such as magnesium stearate, sodium lauryl sulfate and talc may also be added when compressing the tablets. In the case of suspensions and emulsions, the active substances can be mixed not only with the abovementioned excipients, but also with various substances which improve the dye and the colorant.

The active ingredients may be contained in capsules, tablets. pastilles, dragees, ampoules, etc., also in unit amounts, each dosage unit being formed containing a single unit dose of the active ingredient.

The novel compounds may also be present in the formulations in admixture with other known active ingredients.

In general, about 50 to about 300 mg / kg of body weight per day has proven advantageous to achieve good results. Nevertheless, it may be necessary to reduce the amount, depending on the type and body weight of the object to be treated, its individual tolerability of the medicament, the type of dosage form and the administration of the medicament, as well as the time or time interval in which the administration takes place. Thus, in some cases, less than the aforementioned lowest amount may be sufficient, while in other cases the upper limit of this range must be exceeded. In the case of administration of larger amounts, it is sometimes recommended to divide the total daily dose into multiple individual doses.

Claims (1)

OBJECT OF THE INVENTION 1. A process for the preparation of the novel quinazolone diurethanes of the general formula R is C 1 -C 2 alkyl, characterized in that ae 6-isocyanato-2- (3-isocyanatophenyl) -3-phenyl-4- (3 H) -quinazolone of formula II O (II) is reacted with an alcohol of formula III R-OH (III) wherein R is as defined above, optionally in the presence of inert solvents.