FI78468C - FOERFARANDE FOER FRAMSTAELLNING AV TERAPEUTISKT ANVAENDBARA 3,4,5-TRIHYDROXIPIPERIDINDERIVAT. - Google Patents

Description

1 78468

Process for the preparation of therapeutically useful 3,4,5-trihydroxypiperidine derivatives

This invention relates to a process for the preparation of novel 3,4,5-trihydroxy-piperidine derivatives. These new compounds are useful as medicaments, for example in the treatment of precursors of diabetes, peritonitis, constipation, gastrointestinal inflammation, meteorism, flatulence, caries, atherosclerosis, hypertrophic and cardiac disease, and in particular diabetes mellitus, hyperlipidemia. influencing the fat ratio in favor of the meat portion.

The compounds of the invention have

general formula I

OH

| CH-OH H

wherein 25 is -COOR 4, -CO-NR 2 -R 2 or -CH 2 -OR 2, wherein R 1 and R 2 represent hydrogen or a lower alkyl group.

It has been found that the novel compounds of the formula I are effective inhibitors against β-glucosidases, in particular with respect to disaccharides. Thus, the new compounds are valuable substances, affecting a number of metabolic events and thus increase the field of technology for drugs.

Compared to 2-hydroxymethyl-3,4,5-trihydroxypiperidine known from DT-OS 2,656,602, these 35 new compounds have advantageous therapeutic properties.

2 78468

According to the invention, the compounds of the formula I are prepared by:

(a) formula III

OH

5 H0 γ ^ γ-0Η I L · ''01 111 | ^ CH2 OH

B

The 2,6-imino-2-hydroxymethyl-2,6-dideoxy-L-ido- (L-gulo) hexonic acid nitrile of formula 10 is saponified according to formula V

OH

"0 1 ™

Tn> ooh

| L CH2OH H

which is then esterified, if desired, and the ester is reacted with amine 20, if desired, to form an amide or, if desired, reduced with a hydrogen donating reducing agent of formula VI.

OH

HO 10 '25 LJr'0'201' i ^ CH2OH h, or b) CH-, H, C of formula VII. $ 3

35 h2n-h2c and CH90H

OH

The compound of formula VIII is deprotected with a mineral acid to give a compound of formula VIII

3 78468 [0C; -

h2n-h2c I

5 OH

which is reacted directly in solution or after isolation with cyanide of formula III

10 OH

1.0 γ ^ γ0 "LV ™

I t-CH 2 OH

To a compound of formula 15 which is treated according to method a) above to give a compound of formula I.

The starting compounds used are for the most part known or can be easily prepared by methods known per se (vit. H. Paulsen, J. Sangster and K. Heyns, Chem. Ber. 100 (1967), pp. 802-815).

Examples of hydrogen-donating reducing agents which can be used are catalytic hydrogen, alkali metal borohydrides, alkali metal cyanoborohydrides, dialkylaminoboranes or formic acid. In the reductive amination to N-alkylamines, sodium cyanoborohydride is preferably used, hydrogenating the nitrile group to the aminomethyl group, preferably hydrogen in the presence of Raney nickel or in the presence of metal catalysts. The hydrogenation is generally carried out at a hydrogen pressure of 1 to 150 bar and a temperature of between 20 and 150 ° C; in this case, protic, polar solvents, in particular water and al-35 alcohols, are preferably used as solvent.

4 73468

Among the new compounds of the formula I, mention may be made in particular of compounds in which R2 is -COOH, -COCK2H -CH2OH or -CONH2.

The compounds according to the invention are suitable for the treatment of the following conditions: pre-diabetes, peritonitis, gastrointestinal inflammation, flatulence, phthalitis, caries, atherosclerosis, overexertion and in particular hyperlipemia, diabetes and cardiac obesity.

In order to broaden the range of action, it may be advantageous to combine inhibitors of glucose hydrolases which complement each other, whereby the compounds according to the invention can be combined with one another or with previously known inhibitors. In this case, for example, it may be expedient to combine the sucrase inhibitors prepared according to the invention with previously known amylase inhibitors.

In many cases, combinations of compounds of the formula I with known oral antidiabetic agents (β-cytotropic sulphonylurea derivatives and / or blood sugar-acting biguanides) and blood lipid-lowering action zones, such as clofibrate, nicotinic acid and other cholesty acids, are also preferred.

The novel compounds may be administered undiluted, for example as powders or in a gelatin box or together with a carrier, in conventional pharmaceutical compositions which may contain, for example, 0.1 to 99.5% of active ingredient in association with a pharmaceutically acceptable, non-toxic inert carrier, the carrier may contain one or more solid, semi-solid or liquid diluents, excipients and / or non-toxic, inert and pharmaceutically acceptable excipients for the compositions.

5 73468

Although the dosage and dosing schedule of the compounds of formula I must be carefully determined in each case according to a thorough examination by a skilled artisan, taking into account the patient's age, weight 5 and condition, disease quality and severity, dosage is usually in the range of about 0.1 to 100 ml / kg body weight per day. In many cases, a sufficient therapeutic effect is obtained with a lower dose, while in other cases a higher dose is required. Oral administration may be by the use of solid and liquid dosage units such as powders, tablets, pills, capsules, suspensions, solutions, and the like.

In addition to the aforementioned pharmaceutical compositions, foods containing compounds of formula I, e.g. sugar, bread, potato products, fruit juice, beer, chocolate and other confectionery, as well as preserves, such as marmalades, may be prepared by adding a therapeutically effective amount of at least one inhibitors of the invention.

Foods prepared using the compounds of formula I are suitable both for the diet of a patient suffering from metabolic disorders and also for the feeding of healthy individuals as a method of preventing metabolic disorders.

The compounds prepared according to the invention further have the property of strongly influencing the ratio between the proportion of undesirable fat in animals and the proportion of lean meat desired (lean meat) in favor of lean meat. This is particularly relevant for the rearing and keeping of livestock in agriculture, such as pigs for fattening, but is also of great importance for the rearing and keeping of other livestock and livestock 35. Their use can further lead to a significant rationalization of animal nutrition in terms of time, quantity and quality. Because they cause a certain slowing down in digestion, the residence time of the nutrients in the gastrointestinal tract is prolonged, allowing a smaller amount of ad libitum feeding. Furthermore, their use allows in many cases significant savings in valuable protein feed.

The amount of active ingredient administered to animals to obtain the desired effect can be varied within wide limits due to the advantageous properties of the active ingredient. It is preferably about 0.1 mg to 1.0 g, especially 1 to 100 mg of feed per day. The duration of dosing can vary from 10 hours or a day to several years. The appropriate amount of active ingredient and the duration of dosing are closely related to the purpose of feeding. They depend in particular on the species, sex, state of health and purpose of the animal and can be easily determined by a professional.

The active ingredients according to the invention are administered to the animals by conventional methods. The route of administration depends in particular on the species of animal, its behavior and its general condition. Thus, an-20 raising can be performed once or more times a day, at regular or irregular intervals orally. For reasons of convenience, oral dosing is recommended in many cases, especially when watering and feeding animals.

The active ingredients can be administered as such or in preparations, the preparation being understood as meaning both a premix mixed with a non-toxic inert carrier, which may be arbitrarily chosen, and also as part of a complete feedingstuff as an additional feed or compound ingredient in an exclusive compound feed. Dosage of suitable preparations in drinking water can also be used.

The active ingredients can also be administered as preparations in combination with other nutrients and active ingredients, for example mineral salts, trace elements, vitamins, egg whites, energy-giving substances (such as starch, sugar, fats), colorants and / or flavorings or other feed additives such as, for example, 7 78468 with growth promoters in a suitable form. The active ingredients may be administered to the animals before, during or after feeding.

Oral dosing with feed 5 and / or drinking water is preferred, in which case all or part of the active ingredient is added to the feed and / or drinking water as required.

The active ingredients can be added by conventional methods simply by mixing as pure substances, preferably in finely divided form or as preparations with edible, non-toxic carriers, if desired also as a premix or concentrate in feed and / or drinking water. The feed or drinking water may contain the active ingredients according to the invention, for example in concentrations of about 0.001 to 5.0% by weight, in particular 0.02 to 2.0% by weight. The optimum concentration of the active ingredient in the feed or drinking water depends in particular on the amount of feed and / or drinking water used by the animals and can be easily determined by a person skilled in the art.

The premixes may preferably contain from about 0.1 to 5%, in particular from 0.5 to 5.0% by weight, of the active ingredient according to the invention, with arbitrarily selected edible carriers and / or mineral salts, for example carbonated feed lime, and are prepared by customary mixing methods. .

The compound feed preferably contains 0.001 to 5.0, in particular 0.02 to 2.0% by weight, of the active ingredient according to the invention of conventional feed additives, for example cereal flour or by-products, oil cake flour, animal egg white, minerals, trace elements and vitamins. in addition. They can be prepared by conventional mixing methods.

Preferably, the active ingredients are also protected in premixes and compound feed, if desired, by a suitable material covering their surface, for example non-toxic waxes or gelatin, with respect to air, light and / or moisture.

e 78468

Ready-to-use formulation according to the invention A composition example of a feed for poultry contains: 200 g wheat flour 5,340 g maize flour 360.3 g soybean meal 60 g beef 15 g dicalcium phosphate 10 g calcium carbonate 10 4 g iodinated table salt 7.5 g vitamin-mineral mixture active ingredient premix which, when mixed thoroughly, constitute 1 kg of feed.

The vitamin-mineral mixture contains:

15 6,000 I.E. vitamin A

1,000 I.E. vitamin 10 mg vitamin E 1 mg vitamin 3 mg riboflavin 20 2 mg pyridoxine 20 mg vitamin B ^ 2 5 mg calcium pantothenate 30 mg nicotinic acid 200 mg choline chloride 25 200 mg MnSO 4 x H 2 O 140 mg ZnSO 2 x H 2 O 100 mg FeSO 2 x H 2 O and 20 mg CuSO 4 x H 2 O.

The active ingredient premix contains the desired amount of active ingredient prepared according to the invention, for example 1600 mg, in addition to 1 g of DL-methionine and enough soybean meal to give 3.2 g of premix.

9 78468

An example of a compound feed composition for pigs containing a compound of formula I contains: 630 g of feed grain meal (containing 200 g of maize, 150 g of barley, 150 g of oats and 130 g of wheat meal) 5 80 g of fish meal 60 g of soybean meal 58.8 g of tapioca meal 38 g of brewer's yeast 50 g of a vitamin-mineral mixture for pigs (composition 10 same as in chicken feed) 30 g of flax cake flour 30 g of corn glue feed 10 g of soybean oil 10 g of sugar cane molasses and 15 2 g of active ingredient premix (composition similar to chicken feed, for example) giving 1 kg of feed by thorough mixing.

The compound feeds shown are preferably used for rearing and fattening chickens or pigs; however, they may be used as the same or equivalent composition for the rearing or fattening of other animals.

The compounds of the formula I can be used individually or in arbitrary mixtures with one another.

In vitro sucrase inhibition assay The in vitro sucrase inhibition assay allows the determination of the enzyme inhibitory effect of a substance by comparing the effect of a solubilized intestinal disaccharidase complex in the presence and absence of an inhibitor (so-called 100% value).

In this case, practically glucose-free sucrose (glucose <100 ppm) is used as a medium for determining the specificity of the inhibition test; The enzyme activity assay is based on the spectrometric determination of released glucose by glucose dehydrogenase using nicotinamide adenine dinucleotide as a cofactor.

10 78468

The sucrase inhibitory unit (SIE) is defined as the inhibitory effect which, in a given number of tests, reduces the pre-given saccharolytic effect by one unit (sucrase unit = SE); the sucrase unit is then defined by its enzymatic activity which, under predetermined conditions, degrades one pmole of sucrose per minute and thus results in the release of one pmole of sucrose / minute as determined in the test and the release of one pmole of fructose not determined in the experiment.

Intestinal disakkaridaasikompleksi obtained from porcine intestinal mucosa using tryptic digestion; precipitation from 66% ethanol at -20 ° C, the precipitate is placed in 100 mmol of phosphate buffer (pH 7.0) and dialyzed against the same buffer.

10 [mu] l of the sample solution formed so that the extinction of the sample is at least 10% but not more than 25% below 100% is added with 10 [mu] l of a 20 dilution of the intestinal disaccharidase complex in 0.1 mol of maleinate buffer (pH 6, 25) and pre-incubated for 10 minutes at 37 ° C. The dilution of the disaccharidase complex is adjusted to an activity value of 0.1 SE / ml.

The saccharolytic reaction is then started by adding 100 μl of a 0.4 M solution of sucrose (pH 6.25) and after a 20 minute incubation period at 37 ° C and adding 1 ml of glucose dehydrogenase reagent to a flask of glucose dehydrogenase-mutarotase, lyophilized "MERCK 14053") and 331.7 mg / β-nicotinamide adenine dinucleotide (free acid, "BOEHRINGER", purity I) dissolved in 250 ml of 30 Tris buffer, pH 7.6, stop the reaction.

To detect glucose, incubate for 30 minutes at 37 ° C and then perform a photometric measurement at 430 nm against pure reagent (with enzyme but without sucrose).

11 78468

Calculating the inhibitory effect of inhibitors is difficult in that even small changes in the test arrangement, for example a 100% value that varies slightly from one assay to another, already have a noticeable effect on the test result. These difficulties are avoided if a standard is used in each assay. The standard uses a sucrase inhibitor of the formula ^ 25H43 ° 18N, which has a specific inhibitory effect of 77,700 SIE / g and which, when used in an amount of 10 to 20 ng in the test, results in an inhibitory effect of the order of magnitude described above. Knowing the deviations of the extinctions at 340 nm from the 100% value and the inhibitory effect obtained with the standard, the extinction differences can be calculated from the 100% value and the inhibitory effect of the sample solution, taking into account the amount of inhibitor used, by calculating its specific effect in sucrase inhibitory units in grams. places (SIE / g).

Example 1 Preparation of starting material 2,6-imino-2-hydroxymethyl-2,6-dideoxy-L-ido-20 (L-gulo) hexonic acid nitrile

A solution of 46.6 g of 6-amino-6-deoxy-L-sorbofuranose hydrochloride monohydrate was added to 200 ml of 0.5 N hydrochloric acid together with 14.7 g of fiumium cyanide, and the mixture was stirred for 3 hours. The mixture was then evaporated in vacuo at 25 ° C to a thin syrup, 200 ml of methanol / ethanol (1: 1) were added and the separated salt was removed by filtration. The filtrate was evaporated to dryness at 25 ° C in vacuo, the resulting crystalline solid was mixed with ethanol, filtered and washed with ethanol / diethyl ether and diethyl ether. 34.5 g (92% of theory) of colorless crystals with a melting point of 156 DEG C. (dec.) Were obtained.

Rf value 0.194 (eluent 1)

Rf value 0.119 (eluent 2) 35 Rf value 0.6 (eluent 3) i2 78 468

Mobile solution 1 = chloroform / methanol / 25% ammonia solution in a volume ratio of 6: 4: 1

Run solution 2 = chloroform / ethyl acetate / methanol / 25% ammonia solution in a volume ratio of 40: 40: 30: 1.

Example 2 2-Hydroxymethyl-3,4,5-trihydroxypiperidine-2-carboxamide

10 OH

G0-γ ^ γ-0Η I JjvV1 CO-NH2 Η ^ ΟΗ2ΟΗ 15 g of the compound obtained in Example 1 were mixed with 240 ml of ethanol saturated with hydrogen chloride gas for 3 hours at reflux temperature, cooled and suction filtered. The solid product was transferred to a 25 cm long and 3 cm 20 wide column containing the Amberlite cation exchanger IR 120. Washed well with water and then eluted with 0.2% ammonia. Fractions containing pure compound were collected and evaporated to dryness, the oily residue crystallized on trituration with methanol.

2.3 g of colorless crystals were obtained, m.p. 200-202 ° C (dec.). Activity SIE / g 264 181.

Example 3

2-Hydroxymethyl-3,4,5-trihydroxypiperidine-carboxylic acid ethyl ester 30 OH

U / JCOOC2H5

U ^ CH2OH

20 g of the compound obtained in Example 1 were mixed with 240 ml of ethanol saturated with hydrogen chloride for 3 hours at reflux temperature, cooled and suction filtered.

i3 78468

The filtrate was evaporated to dryness and the residue was purified on a 120 cm long and 6 cm wide cellulose column to give the compound as the hydrochloride eluting with 90% acetone. The resulting hydrochloride was applied to an Am-5 berlite IR 12O cation exchange column, washed well with water and then eluted with 0.5% ammonia. Fractions containing pure compound were combined and evaporated to dryness. The evaporation residue crystallized slowly from acetone with the addition of a little ethanol. 5.8 g of 10 colorless crystals were obtained; mp. 106-108 ° C.

Example 4 2,2-Dihydroxymethyl-3,4,5-trihydroxypiperidine

OH

° H

15 L Jh CH2 ° h

H 2 CH 2 OH

1.2 g of the compound obtained in Example 3 was dissolved in 20 ml of absolute methanol, and 2 g of sodium borohydride was added portionwise to the mixture. The solution was stirred for 2 hours, carefully acidified with dilute hydrochloric acid and evaporated to dryness. The evaporation residue was dissolved in about 200 ml of a methanol / water mixture (1: 1) and applied to an Amberlite IR 120 cation exchange column. Washed 25 well with water. The product was eluted with 2% ammonia and the resulting solution was evaporated to dryness, the oily evaporation residue was hot dissolved in a small amount of methanol and allowed to stand. After crystallization, the crystals were filtered off with suction and washed with methanol. 0.6 g of colorless crystals were obtained; 30 sp. 148-150 ° C.

Claims (2)

The 2,6-imino-2-hydroxymethyl-2,6-dideoxy-L-ido- (L-gulo) hexonic acid nitrile according to CH 2 OH H 25 is saponified to the formula VOH! ^ OH 30. jCOOH V ·. to a compound of formula VI 78468 OH hoyVoh [^ Ν ip ^ CH2OH VI 5. ch2oh H, which is then esterified with an amine or, if desired, reduced with an amine to give an amide or, if desired, reduced with a hydrogen-donating reducing agent. , or b) the compound of formula VII is deprotected with a mineral acid to give a compound of formula VIII to give a compound of formula VIII. HX | \ _1 / lch2oh A compound of the formula H2N-H2C I OH which is reacted directly in solution or after isolation with cyanide to give a compound of the formula 35 m 16 73 468 OH HOn 1 ^ OH LyCN III \ N 5. i-CH2OH H to be treated according to method a) above to give a compound of formula I. i7 78468 For the preparation of a therapeutically active 3,4,5-trihydroxypiperidine derivative in the form of all-5 forms of OH η ° ^ Λ ° η L 10 I ch9oh H color 1 * 2 is -COOR ^, -CO-NR ^ R 2 or -CH 2 -OR 2, colors R 2 and R 2 are optionally selected from the group consisting of alkyl, the like, and a) 2,6-imino-2-hydroxymethyl-2,6-dideoxy-L-ido - (L-gulo) hexonsyranitrile with formula III 20 ° H HO .JyOH L ^ CN hi | L ch2oh H 25 account for the formation of the form V OH hovVob C> cooh | ^ ClI2OH H vilken därefter, om sä volunskas, förestras ocst ester Brin-35 gas att reagera med en Amin account en amid ell reducer, om é selfskas, med ett reducerande medel, vilket avger väte, account en förening med Formeln VI