CS251099B2 - Method of new therapeutically applicable 1,2,-5,6-dianhydrohexitols production - Google Patents

Abstract

The invention relates to a process for the production of new ones, therapeutically useful 1,2,5,6-dianhydrohexitols free carboxylic acid groups of 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, a C4 -C8 alkylcarbonyl group which also contains a free carboxyl group or a C2 -C5 alkoxycarbonyl group, a C7 -C10 aralkylcarbonyl group, optionally containing a carboxyl group, or an alkylcarbonyl group of 1 as well as a process for producing salts of these compounds having tumor inhibitory activity. These compounds can be formulated into pharmaceutical compositions containing the novel compounds as active ingredients.

The novel compounds of formula (I) may be prepared by reacting 1,2,5,6-dianhydrohexitol (X) with a hexitol backbone of dulcitol or mannitol.

231099

I o cu '(

CH-OH 1 ch-qh

I m

I ^{0 CH} wherein hexitolovým skeleton is dulcitol, mannitol or with anhydrldem dicarboxylic acid of formula XI

where

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 converting the compound of formula I thus obtained into its salt.

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

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

The didesoxydihalogen derivatives of the above dianhydro compounds can be obtained by treating dianhydrohexitol derivatives containing free carboxyl groups with potassium bromide and hydrogen bromide. Dihalogenated derivatives are crystalline substances that are easy to identify and form in the reaction in a yield of nearly 100 percent.

The dianhydrodisuccinylhexitols prepared according to the process of the invention are relatively poorly water-soluble, which applies even more to dihalogenatedoxydisuccinylhexitols. In order to increase the water solubility of these compounds, they can be converted into salts which can be formed with alkali metals, alkaline earth metals and organic bases.

Compared to known dianhydrohexitols and didesoxydihalogenhexitols, which are unsubstituted or carry only hydrophobic substituents, the novel dianhydrohexitols and didesoxydihalogenhexitols produced by the process of the invention and containing a free carboxyl group have far more beneficial therapeutic effects. To demonstrate this, the properties were compared

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- (4-carboxypropionyl) -dulcitol (diSuDAD) produced by the method of the invention The data obtained when used in the cases of Walker's im carcinosarcoma, S-180 sarcoma and P388 leukemia are shown in Table 1. .

Z 1 Z 3 9

T a b u 1 k a 1 Compound LD 50 LDio EDso EDso mg / kg mg / kg , µM / 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 have been described in Institóris, L. et al.: Arzenim.-Forsch. 17, 1945-1955, 1967. It is also known that the therapeutic index of 1,2,5,6-diauhydrodulcitol derivatives acetylated at position 3 and / or 4 is favorable for most transplanted tumors as compared to non-acetylated compounds as described in MA-2630/1974, 12, 24, Otvos, L., Elekes, I, Tydrahedron Letters 29, 2477-2480, 1975, Somfai-Relle, S., et al., Proc. 10 Ing. Cong. Chemother, II. 1302-1304 (1977). A further substantial improvement in the therapeutic index is seen when the acyl group contains a free carboxyl group.

16 4

111 15 7

550 14 40

In order to characterize the antitumor effect of terminally bifunctional 3,4-bis-O-acylhexitols containing free carboxyl groups, the therapeutic index was compared on transplanted tumors, the 1,2,5,6-dianhydrO'-3,4- index was compared. bis - ((3-carboxypropionyl) dulcitol (disuDAD) with the 1,2,5,6-dianhydro-3,4-bis-O-acetyldulcitol index (diacDAD), which is the most characteristic of 1,2,5,6 -dianhydro-3,4-diacylhexitols containing no free carboxyl group The experiments were carried out under the same conditions.

Acute toxicity

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

Table

compound test animal gender LD50 mg / kg diacDAD swiss males i. 34 neinbrední males p. 43 mice males i. 39 29 disuDAD swiss males i. neinbrední males p. 660 mice males i. 505 diacDAD neinbrední males i. -950 rats 28 Wistar disuDAD neinbrední males i. 220 rats males p. <200 Wistar 2) Ascites in P388 leukemia in mice the normal dose which does not cause death based on toxicity and causes only a decrease One million ascites cells were inoculated animal weight. well to BDFi male mice. The mice were Results with single intraperitone- treated intraperitoneally either daily after Administration: either 9 days from the first day for transplantation, or one day following the trans- diacDAD disuDAD plantation. The evaluation is based on extension of 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 breadth of disuDAD is higher than the therapeutic breadth of diacDAD for single and especially long-term administration.

3) Sarkom S180

Sarcoma S180 was inoculated under the skin of Swiss male mice as a small portion of the tissue. Intraperitoneal administration of the drug was performed from the following day, the vessels were removed for day 11 evaluation.

Results after single intraperitoneal administration on Day 1 for tumor transplantation.

diac DAD ED 50 - 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 / ED 50 = 7.5

Intravenous use

Intravenous administration of disuDAD is particularly preferred. When the active ingredient is dissolved in IM sodium bicarbonate, it can be administered intravenously at any concentration.

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

DisuDAD by intravenous administration has a good inhibitory effect on s180 sarcoma in transplantation under the skin.

Results for the treatment of subcutaneous sarcoma S180 by 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 every day from the day following the transplantation, the administration time is 14 days. The effect is evaluated by weighing the tumors 21 days after transplantation.

diacDAD ED 50 = 14 x 3 mg / kg

MTD = 14 x 7 mg / kg Th1 = MTD / ED 50 = 2.3 disuDAD ED 50 = 14 x 10 mg / kg

MTD = 14 x 50 mg / kg Th1 = MTD / ED50 = 5.0

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

5) Walker 256

The Walker tumor is inoculated with 2 × 10 ⁷ ascitic tumor cells intramuscularly into the thigh of male Wistar rats.

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

A quantitative comparison of diacDAD and disuDAD was performed on the basis of a blood count performed on day 4 based on the tumor inhibitory effect found by weighing the tumor on day 10.

Labeling doses of diacDAD compared to those 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 on the basis of the marking doses

Table compound diacDAD disuDAD therapeutic index

LD10 / ED50 security index

Grtox5o / ED90

Lytox5o / ED90

Hematological. index

Lytox5o / Grtox50

Toxic, non-lethal area

LDio / Grtox50

LDio / Lytoxos

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

They may also be converted into solutions or suspensions for injection using water or a physiologically inert organic solvent such as various glycols and the like. The solutions obtained can be administered, for example, intravenously, intramuscularly, intraperitoneally or in various cavities, and can contain 1 to 10% of active ingredient, suspensions can be administered into muscle, intraperitoneally or in cavities, and contain, for example, 1-70% of active ingredient.

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

The pharmaceutical preparations may contain a single active compound of the formula I or II or a mixture thereof.

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 with succinic anhydride, 1.46 g (10 mmol of 1,2,5,6-dianhydrodulcitol) are 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 stirring, and the mixture is heated to 45 [deg.] C. and stirred at this temperature for 8 hours. The residue was mixed with ethyl acetate / water and the resulting mixture was carefully acidified with ice-cooling to pH 4-5. The organic phase was separated, washed with water, dried and evaporated to give 2.5 g of a solid residue. which corresponds to the quantitative yield when calculated as monosuccinyldianhydroducitol.

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

Example 2

Process for the preparation of 1,2,5,6-dianhydro-3,4-diphenyldulcitol with 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 under magnetic stirring, and then 5 ', 6 ml (40 millimoles) of triethylamine are added, followed by 5.92 g (40 mmol) of phthalic anhydride powder was heated to 55 ° C and stirred at this temperature for an additional 90 minutes. The deposited white material containing diphthalyltriethylamine and monophthaltriethylamine was separated from the benzene phase.

Trituration of this white solid with petroleum ether gave the non-hygroscopic salt. Melting point 138-140 ° C.

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

The resulting crude material is chromatographed on a silica gel column using a 1: 1 mixture of methanol and ethyl acetate. The pure product fractions were combined, the solutions were evaporated in vacuo and the resulting solid was recrystallized from ether / petroleum ether. 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 are thus obtained.

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

The product is homogeneous in thin layer chromatography. R _f = 0.8 in ethyl acetate: methanol 1: 1 p-nitrobenzylpyridinu for detecting the substance. Melting point 170-172 ° C.

Infrared light has bands at

550-2,800 cm ^-1 (acylhydroxyl),

1725 cm ^-1 (estercarbonyl),

680 cm ^-1 (carbonyl acids),

585 and 1562 cm ¹ (aromatic ring).

The characteristic band for saccharhydroxyl groups at 3400 cm ^-1 cannot be demonstrated.

Analysis for:

C22H18O20, molecular weight:

442.38, calculated:

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

% H, 4.06;% O, 35.98.

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

The product is homogeneous in thin layer chromatography. R _f = 0.4 in the above said mixture.

Melting point: 79-81 ° C.

The spectrum in infrared light is the same as that for the diphthyl derivative, but at 3,400 cm ^{-1 it} contains a characteristic band for the saccharide hydroxyl groups.

Analysis for:

C14H14O5, calculated:

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

63.96 ° C, 5.18% H, 30.15% O.

Example 3

Process for the manufacture of ampoules containing a lyophilized powder, 10 mg of 1,2,5,6-dianhydro-3,4-bis ((3-carboxypropionyl) mannitol)

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

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

The pharmaceutical compositions are obtained as follows:

g of pure (100%) 1,2,5,6-dianhydro-3,4-bis ((S-carboxypropionyl) mannitol, suitable for pharmaceutical use, is placed in a 10 l calibrated container, this active substance dissolves at 20 ° C in distilled water for injection purposes, carbon dioxide-free, and volume is made up to the mark with distilled water of the same quality, then sterilized by filtration under aseptic conditions. The frozen substance is sublimed at hypoeutectic temperature, the final drying is carried out at 30 DEG C. The ampoules are then stoppered with rubber stoppers and sealed with a metal band under sterile conditions and labeled. contain 2 ml of an aqueous solution of sorbitol at a concentration of 15% w / v, the solution being prepared in a conventional manner.

Example 4

Process for preparing capsules containing 100 mg of 1S, 5'-dianhydro-5'-bis-4'-carboxypropionyl imannitol

Each capsule contains:

100 mg 1,2,5,6-dianhydro-3,4-bis - (R-carboxypropionyl) mannitol mg Carbowax 6000 and mg talc.

The carbowax and talc are homogenized and the resulting finely-powdered homogenate is mixed with the pure crystalline derivative produced by the process of the invention and the granule can thus be filled into hard gelatin capsules. Each capsule contains 110 mg of granular mixture.

Example 5

Process for the preparation of capsules soluble in the intestine

The capsule prepared according to the method of Example 3 is provided with a coating which 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.