CS251099B2 - A method for producing novel therapeutically useful 1,2-5,6-dianhydrohexitols - Google Patents

Abstract

The invention relates to a method for producing new, therapeutically useful 1,2,5,6-dianhydrohexitols containing free carboxyl groups of the general formula I

Description

R represents an alkylcarbonyl group of 4 to 8 carbon atoms, which also contains a free carboxyl group and/or an aralkylcarbonyl group of 7 to 10 carbon atoms, optionally containing a carboxyl group,

R ¹ represents a hydrogen atom, an alkylcarbonyl group of 4 to 8 carbon atoms, which also contains a free carboxyl group or an alkoxycarbonyl group of 2 to 5 carbon atoms, an aralkylcarbonyl group of 7 to 10 carbon atoms, optionally containing a carboxyl group, or an alkylcarbonyl group of 1 to 3 carbon atoms, as well as a process for preparing salts of these compounds which have an inhibitory effect on tumors. These substances can be processed into pharmaceutical compositions which contain the above-mentioned novel compounds as an active ingredient.

The process according to the invention allows the preparation of new compounds of general formula I by reacting 1,2,5,6-dianhydrohexitol of general formula X, the hexitol skeleton being dulcitol or mannitol,

231099

I'm sorry (

CH — OH I ch-qh

I am

I ^{0 CH} Í where the hexitol skeleton is dulcitol or mannitol, with a dicarboxylic acid anhydride of the general formula XI

where

A represents an alkylene group of 2 to 10 carbon atoms or a phenylene group, preferably in an anhydrous environment in the presence of a base and optionally converting the compound of general formula I thus obtained into its salt.

The β-carboxypropionyl derivatives can be obtained by the process according to the invention by directly acylating the corresponding dianhydrohexitols with succinic anhydride.

Preferably, the 1,2,5,6-dianhydrodulcitol is reacted with succinic anhydride in an anhydrous solvent in the presence of a tertiary base. The product obtained, isolated from the reaction mixture in almost quantitative yield, is then subjected to chromatography on a silica gel column to separate the monosuccinyl and disuccinyl derivatives. These products are identical to those obtained by hydrogenolysis of the corresponding benzyl ester.

The dideoxydihalogen derivatives of the above-mentioned dianhydro compounds can be obtained by treating dianhydrohexitol derivatives containing free carboxyl groups with potassium bromide and hydrogen bromide. The dihalogen derivatives are crystalline substances that are easy to identify and are formed in the reaction in a yield of almost 100 percent.

The dianhydrodisuccinylhexitols prepared by the process of the invention are relatively poorly soluble in water, which is even more true for the dihalodesoxydisuccinylhexitols. In order to increase the solubility of these compounds in water, they can be converted into salts which can be formed with alkali metals, alkaline earth metals and organic bases.

Compared to known dianhydrohexitols and dideoxydihalohexitols, which are unsubstituted or carry only hydrophobic substituents, the new dianhydrohexitols and dideoxydihalohexitols prepared by the method according to the invention and containing a free carboxyl group have much more advantageous therapeutic effects. In order to demonstrate this fact, the properties of the

1.2.5.6- dianhydrodulcitol (DADj and 1,2,5,6-dianhydro-3,4-diacetyldulcitol (dlAcDAD), which are known from the literature with properties

1.2.5.6- dianhydro-3,4-bis-(^-carboxypropionyl)dulcitol (diSuDAD), produced by the method of the invention. The data obtained when used in cases of Walker i. m. carcinosarcoma, S-180 sarcoma and P388 leukemia are shown in Table 1.

From 1 to 3 9

T a b l e 1 k a 1 Compound LDso LDio EDso EDso mg/kg mg/kg ,uM/kg μΜ/kg

DAD 15 10 diAcDAD 34 25.5 diSuDAD 630 190

The novel acylated dianhydrohexitol compounds produced by the process of the invention can be used to inhibit tumors in various ways.

It is known that the cytostatic effect of hexitol with two terminal functions can usually be favorably influenced by acylation of the secondary hydroxy group. The results of these tests were described in Institóris, L. et al.: Arzenim.-Forsch. 17, 1945 to 1955, 1967. It is also known that the therapeutic index of 1,2,5,6-diauhydrodulcitol derivatives, acetylated in position 3 and/or 4, is favorable in the case of most transplanted tumors compared to non-acetylated compounds, as described in MA-2630/1974, 12. 24.: Otvos L., Elekes I.: Tydrahedron Letters 29, 2477 to 2480, 1975.: Somfai-Relle S. et al.: Proc. lOth Ing. Cong. Chemother sv, II. 1302-1304, 1977. A further substantial improvement in therapeutic index can be observed when the acyl group contains a free carboxyl group.

16 4

110 15 7

550 14 40

In order to characterize the antitumor effect of terminally bifunctional 3,4-bis-O-acylhexitols containing free carboxyl groups, a comparison of the therapeutic index on transplanted tumors was performed, comparing the index of 1,2,5,6-dianhydro-3,4-bis-((3-carboxypropionyl)dulcitol (disuDAD) with the index of 1,2,5,6-dianhydro-3,4-bis-O-acetyldulcitol (diacDAD), which is the most characteristic substance from the group of 1,2,5,6-dianhydro-3,4-diacylhexitols that do not contain any free carboxyl group. The experiments were performed under the same conditions.

Acute toxicity

After a single injection, the animals were monitored for 21 days. The results are shown in the following table.

Table

compound experimental animal gender route of administration LDso in mg/kg diacDAD Swiss males i. p. 34 non-inbred males p. o. 43 mice males i. v. 39 29 disuDAD Swiss males i. p. non-inbred males p. o. 660 mice males i. v. 505 diacDAD non-inbred males i. p. -950 rats 28 Wistar disuDAD non-inbred males i. p. 220 rats males p. o. <200 Wistar 2) Ascites in P388 Leukemia in Mice small dose that does not cause death based on toxicity and only causes a decrease One million ascites cells were inoculated- the weight of the animal. to male mice of the BDFi strain. The mice were Results of a single intraperitoneal treated intraperitoneally either daily after official submission: or 9 days from the first day for transplantation, or one-time the day after the trans- diacDAD disuDAD plantation. The evaluation is based on the pro- extending life expectancy. MED: 11.5 mg/kg MED: 100 mg/kg MTD: 16.0 mg/kg MTD: 200 mg/kg MED = minimum effective dose T/C % 130 MTD/MED = 1.4 MTD/MED = 2.0

MTD = maximum tolerated dose; maxi251399

The therapeutic range of disuDAD is higher than that of diacDAD for single administration and especially for long-term administration.

3) Sarcoma S180

The S180 sarcoma was inoculated subcutaneously into male Swiss mice as a small piece of tissue. Intraperitoneal administration of the active substance was performed from the following day, the vessels were removed for evaluation on day 11.

Results after a single intraperitoneal administration on day 1 for tumor transplantation.

diac DAD EDso — 1 x 8.7 mg/kg

MTD = 1 x 21 mg/kg

MTD/ED 50 = 2.4 disuDAD MD 50 = 1 x 40 mg/kg

MTD = 1 x 300 mg/kg

MTD/ED50 = 7.5

Intravenous administration

Intravenous administration of disuDAD is particularly advantageous. If the active substance is dissolved in sodium bicarbonate at an IM concentration, it can be administered intravenously at any concentration.

The clinical use of diacDAD is limited by the incomplete solubility of this substance, so the compound can only be administered as an intravenous infusion.

DisuDAD when administered intravenously has a good inhibitory effect on S180 sarcoma when transplanted under the skin.

Results of treatment of subcutaneous sarcoma S180 with intravenous administration:

diacDAD ED84 = 2 x 13 mg/kg

LD50/ED84 = 1.5 disuDAD ED84 = 1 x 400 mg/kg x 210 mg/kg

LD50/ED84 = 2.4

4) Harding-Passey melanoma

A small portion of Harding-Passey tumor tissue is transplanted under the skin of male mice. The active substance is administered daily from the day after transplantation for 14 days. The effect is evaluated by weighing the tumors 21 days after transplantation.

diacDAD ED50 = 14 x 3 mg/kg

MTD = 14 x 7 mg/kg Thl = MTD/EDso = 2.3 disuDAD EDso = 14 x 10 mg/kg

MTD = 14 x 50 mg/kg Thl = MTD/ED 50 = 5.0

DisuDAD has a very favorable therapeutic index in the case of slow-growing refractory Harding-Passey melanoma.

5) Walker 256

Walker tumor is inoculated with 2 χ 10 ⁷ ascites tumor cells intramuscularly into the thigh of male Wistar rats.

This tumor is relatively well treated and is suitable for monitoring the dose-response of a single intraperitoneal administration, as well as for monitoring the effect of these doses on the blood count.

Quantitative comparison of diacDAD and disuDAD was performed based on blood counts performed on day 4 based on the tumor inhibitory effect detected by tumor weighing on day 10.

Labeling doses of diacDAD compared to these doses of disuDAD

Table Compound dose EDso mg/kg granulocytes ÍOX50 mg/kg lymphocytes toxso mg/kg LDto mg/kg diacDAD 3.0 10.0 24.0 25.5 disuDAD 5.0 20.0 110.0 190.0

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Therapeutic index, calculated based on marker doses

Table compound diacDAD disuDAD therapeutic index

LDio/EDso safety index

Grtox5o/ED90

Lytox5o/ED90

Hematological index

Lytox5o/Grtox50

Toxic, non-lethal area

LDio/Grtox50

LDio/Lytoxos

The new compounds can be converted into pharmaceutical preparations suitable for oral administration, for example into tablets as such or in admixture with conventional excipients such as starch, lactose, talc, etc. Tablets for oral administration can contain, for example, 20 to 90% of the active ingredient.

These substances can also be converted into solutions or suspensions for injection purposes using water or a physiologically inert organic solvent, for example various glycols and the like. The solutions obtained can be administered, for example, intravenously, intramuscularly, intraperitoneally or into various cavities and can contain 1 to 10% of the active substance, suspensions can be administered intramuscularly, intraperitoneally or into cavities and can contain, for example, 1 to 70% of the active substance.

The compounds of the invention can also be administered topically to the skin or to surgical scars as such or in admixture with conventional antibacterial and/or healing agents such as sulfonamides, corticosteroids, vitamins and the like.

Pharmaceutical preparations may contain a single active ingredient of general formula I or II or a mixture of these ingredients.

The invention will be illustrated by the following examples, all starting materials used in these examples are known.

Example 1

Acylation of 1,2,5,6-dianhydroprodulcitol by the action of succinic anhydride. 1.46 g (10 mmol) of 1,2,5,6-dianhydrodulcitol is dissolved in 20 ml of anhydrous ethyl acetate, and 5 ml of pyridine is added to the solution.

8.5 38

3.3 4.0

8.0 22.0

2.4 5.5

2.6 9.5

1.1 1.7 and then 2.0 g (20 mmol) of succinic anhydride are added in small portions with constant stirring. The mixture is heated to 45 °C and stirred at this temperature for a further 8 hours. The solvent is distilled off in vacuo, the residue is mixed with a mixture of ethyl acetate and water and the resulting mixture is carefully acidified with ice cooling to pH 4 to 5. The organic phase is separated, washed with water, dried and evaporated. In this way 2.5 grams of solid residue are obtained, which corresponds to the quantitative yield if calculated as monosuccinyldianhydroducitol.

The resulting crude product is chromatographed on a silica gel column using a 1:1 volume mixture of methanol and ethyl acetate. The fractions containing the pure product are combined, the solution is evaporated in vacuo and the resulting solid is recrystallized from a mixture of methanol and ether. In this way, 1.8 g of 1,2,5,6-dianhydro-3-([beta]-carboxypropionyl]dulcitol and 0.4 g of 1,2,5,6-dianhydro-3,4-bis-(carboxypropionyl)dulcitol are obtained. The physical constants of these compounds are identical to the physical constants of the resulting products obtained in Examples 2 and 3.

Example 2

Process for producing 1,2,5,6-dianhydro-3,4-diphthalyldulcitol from 1,2,5,6-dianhydro-3-phthalyldulcitol

2.92 (20 mmol) of 1,2,5,6-dianhydrodulcitol are suspended in 200 ml of anhydrous benzene while stirring with a magnetic stirrer. Then 5.6 ml (40 mmol) of triethylamine are added to the suspension and then 5.92 g (40 mmol) of powdered phthalic anhydride. The mixture is heated to 55 °C and stirred at this temperature for a further 90 minutes. The precipitated white material, which contains diphthalyltriethylamine and monophthaltriethylamine, is separated from the benzene phase.

Trituration of this white solid with petroleum ether gives a non-hygroscopic salt, melting point 138-140°C.

The desired carboxylic acid derivatives are liberated from the above salt by dissolving the salt in a mixture of ethyl acetate and water and carefully acidifying it to pH 4-5 under ice-cooling. The organic phase is separated, washed with water, dried and evaporated. In this way, 4.2 g of a solid residue are obtained, which is a mixture of monophthalyldianhydrodulcitol and diphthalyldianhydrodulcitol.

The resulting crude material was chromatographed on a silica gel column using a 1:1 mixture of methanol and ethyl acetate. The fractions containing pure product were pooled, the solutions were evaporated in vacuo and the resulting solid was recrystallized from a mixture of ether and petroleum ether. In this way, 0.75 g (22%) of 1,2,5,6-dianhydro-3-phthalyldulcitol and 1.33 g (31%) of 1,25,6-dianhydro-3,4-diphthalyldulcitol were obtained.

Physical properties for 1,2,5,6-dianhydro-3,4-diphthalyldulcitol

The product is homogeneous by thin layer chromatography. R _f = 0.8 using a 1:1 mixture of ethyl acetate and methanol and p-nitrobenzylpyridine for the detection of this substance. Melting point 170-172 °C.

Light in the infrared has bands at

550 — 2,800 cm ^-1 (acyl hydroxyl),

1725 cm“ ¹ (ester carbonyl),

680 cm ^-1 (carbonyl acids),

585 and 1,562 cm ¹ (aromatic ring).

The characteristic band for saccharide hydroxyl groups at 3,400 cm- ¹ cannot be demonstrated.

Analysis for:

C22H18O20, molecular weight:

442.38, calculated:

59.73% C, 4.10% H, 36.17% O; found:

59.03% C, 4.06% H, 35.98% O.

Physical properties for 1,2,5,6-dianhydro-3-phthalyldulcitol

The product is homogeneous by thin layer chromatography. R _f = 0.4 using the above mixture.

Melting point: 79 to 81 °C.

The infrared spectrum is the same as that for the diphthalyl derivative, but contains a characteristic band at 3,400 cm“ ¹ for carbohydrate hydroxyl groups.

Analysis for:

C14H14O5, calculated:

64.12% C, 5.34% H, 30.53% O; found *

63.96 °/o C, 5.18% H, 30.15% O.

Example 3

Method for manufacturing ampoules containing lyophilized powder, 10 mg of 1,2,5,6-dianhydro-3,4-bis ((3-carboxypropionyl) mannitol

One ampoule of powder contains 10 mg of 1,2,5,6-dianhydro-3,4-bis(1-carboxypropionyl)mannitol.

- One ampoule of solution contains 300 mg of sorbitol and distilled water for injection up to 2 ml.

Pharmaceutical compositions are obtained in the following manner:

g of pure (100!%) 1,2,5,6-dianhydro-3,4-bis ((S-carboxypropionyl) mannitol, suitable for pharmaceutical purposes, is placed in a calibrated container with a capacity of 10 liters, this active substance is dissolved at a temperature of 20 °C in distilled water for injection purposes, free from carbon dioxide, and the volume is made up to the mark with distilled water of the same quality. The solution is then sterilized by filtration under aseptic conditions. Each 2 ml of the sterile solution thus obtained is filled into ampoules and lyophilized in the usual way. The frozen substance sublimes at a hypoeutectic temperature, final drying is carried out at a temperature of 30 °C. The ampoules are then stoppered with rubber stoppers and sealed with a metal strip under sterile conditions, after which they are labeled. The ampoules containing the powder are packaged together with ampoules containing the solvent, which contain 2 ml of an aqueous solution of sorbitol with a concentration of 15% w/v. %, the solution is prepared in the usual way.

Example 4

Method for manufacturing capsules containing 100 mg of 1,5-dihydro-5-bis-carboxypropionylmannitol

One capsule contains:

100 mg 1,2,5,6-dianhydro-3,4-bis-([r-carboxypropionyl] mannitol mg Carbowax 6,000 and mg talc.

Carbowax and talc are homogenized and the resulting finely powdered homogenate is mixed with the pure crystalline derivative produced according to the process of the invention and the granules thus obtained are filled into hard gelatin capsules. Each capsule contains 110 mg of the granulated mixture.

Example 5

Method of manufacturing enteric-coated capsules

Capsules prepared according to the method of Example 3 are provided with a coating that dissolves only in the intestine.

Claims (2)

SUBJECT VYNALEZU A process for the preparation of novel therapeutically useful 1,2,5,6-dianhydrohexitols of formula I CH CH CH-OH AND CH-OH where CH-OR ¹ 1 CH-Q-R ' CH \ O / ^CH 2 0) CH. CH 7 (x) wherein the hexitol skeleton is dulcitol or mannitol, with a dicarboxylic anhydride of formula XI The CO backbone of hexitol is dulcitol or mannitol, -Wi R represents an alkylcarbonyl group of 4 to 8 carbon atoms which also contains a free carboxyl group and / or an aralkylcarbonyl group of 7 to 10 carbon atoms, optionally containing a carboxyl group, R ¹ represents a hydrogen atom, an alkylcarbonyl group having 4-8 carbon atoms which also contains a free carboxy group or an alkoxycarbonyl group having 2 to 5 carbon atoms, an aralkylcarbonyl group having 7-10 carbon atoms, optionally containing a carboxyl group or an alkylcarbonyl group having 1 to And the salts of these compounds, characterized in that the 1,2,5,6-dianhydrohexitol of formula (X) is reacted: A represents a C 2 -C 10 alkylene group or a phenylene group, preferably in an anhydrous medium in the presence of a base, and optionally converts the compound of formula I thus obtained to its salt. 2. The process of claim 1 wherein the reaction is carried out in an anhydrous environment in the presence of a base.