DK157012B - Analogy process for preparing 3,4,5-trihydroxypiperidine derivatives - Google Patents

Description

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DK 157012 B

The present invention relates to an analogous method for the preparation of novel derivatives of 3,4,5-trihydroxypiperidine which is a useful agent for the treatment of pre-diabetes, gastritis, constipation, gastrointestinal tract infections, meteroism, intestinal air, caries, arteriosclerosis, hypertension and especially diabetic, hyperlipaemia and obesity, and for feeding animals to affect the fat-meat ratio in favor of the amount of meat.

The compounds prepared by the process of the invention have the formula

HO? H OH

Wherein R 2 represents -C00H, -COOR 3 or -CH 2 -OH, and R 3 represents a lower alkyl group.

It has been found that the novel compounds 20

of formula I are active inhibitors of α-glucosidases, especially of disccharidases. Therefore, the novel compounds are valuable agents for influencing many metabolic processes and constitute enrichment in the pharmaceutical field. IN

25 in relation to that of German publication no.

2,656,602 known 2-hydroxymethyl-3,4,5-trihydroxypiperidine are the novel compounds possessing beneficial therapeutic properties.

The process of the invention is peculiar to 3Q in that 2,6-imino-2-hydroxymethyl-2,6-didesoxy-L-ido (L-gulo) hexonic acid nitrile of the formula

HO JP OH

UC

g 2 * CH 2 OH

35 2

DK 157012 B

is hydrolyzed to a compound of formula OH

HVVH

5 CkC00H

H Δ which is then optionally esterified or optionally reduced by hydrogen-donor reducing agents to a compound of formula

BO f ° H

WE

H 2 CH 2 OH 15

The starting compounds used are known to the majority or may be prepared by methods known per se, cf. H. Paulsen, J. Sangster and 20 K. Heyns, Chem. Ber. 100, 802-815 (1967).

As hydrogen-donor reducing agents, e.g. catalytic hydrogen, alkali metal borohydrides, alkali metal cyanoborohydrides, dialkylaminoboranes or formic acid are used. Hydrogenation is usually carried out at pressures of 1-150 bar of hydrogen and temperatures between 20 and 150 ° C, as solvents therefor preferred are polar, polar solvents, especially water and alcohols.

The inhibitors prepared by the process according to the invention are suitable as treating agents in the following indications:

Pre-diabetes, gastritis, constipation, gastrointestinal infections, meteorism, intestinal air, caries, arteriosclerosis, hypertension and especially obesity, diabetic and hyperlipemia.

35 In order to broaden the spectrum of action, it is recommended to combine inhibitors of glycoside hydrolases that mutually complement each other's effect, whatever it is Ο

DK 157012B

3 is about combinations of inhibitors made by the free-range food according to the invention or about their combination with already known ones. Thus it can be e.g. It is useful to combine sucrose inhibitors prepared by the process of the invention with already known amylase inhibitors.

In many cases, combinations of inhibitors prepared by the method of the invention with known oral antidiabetic agents (0-cytotropic sulfonyl-urea derivatives and / or blood sugar-active biguanides) are also advantageous, as well as combinations with blood lipid-reducing agents such as clofibrate, nicotinic acid, cholesterol. .

The compounds can be used without dilution, e.g.

15 as a powder or in a gelatin sheath or in combination with a carrier in a pharmaceutical composition.

Pharmaceutical compositions may contain a greater or lesser amount of the inhibitor, e.g. 0.1-99.5%, combined with a pharmaceutically acceptable non-toxic, inactive carrier, the carrier may contain one or more solid, semi-solid or liquid diluents, fillers and / or non-toxic inactive pharmaceutically acceptable excipients. Such pharmaceutical compositions are preferably in the form of dosage units, i.e.

25 physically separate units containing a certain amount of inhibitor and corresponding to a fraction or multiple of the dose necessary to provide the desired inhibitory effect. The dosage units may contain 1, 2, 3, 4 or more single doses or 1/2, 1/3 or 30 1/4 of a single dose. Preferably, a single dose contains a sufficient amount of active ingredient to obtain an inhibitory effect desired by administration of one or more dosage units according to a predetermined dosage regimen, in which an entire, one-half or one-third, or one-quarter of the daily dose is administered at each head. - and middle-

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DK 157012 B

4 meals.

Other therapeutic agents can also be used. Although the dosage and dosage schedule in each case must be carefully weighed in accordance with a thorough professional judgment and taking into account the patient's aids, weight and condition and the nature and severity of the disease, the dose will usually be in an interval between about .

0.1 to approx. 100 mg / kg body weight per day. day. In many cases, a sufficient therapeutic effect 10 with a smaller dose will thereby be obtained, while in other cases a larger dose is required.

Oral administration may be effected using solid and liquid dosage units such as e.g. with powders, tablets, dragees, capsules, granules, suspensions, solutions and the like.

Dosage prescriptions can be indicated on the capsule.

In addition, the dosage can be assured so that the active substance is delivered protracted, e.g. in that the active substance is contained in polymeric substances, waxes and the like.

The inhibitors can be used individually or in mixtures of several as desired.

Sucrose inhibitor assay in vitro

The in vitro saccharase inhibitor test allows determination of the enzyme inhibitory effect of a substance by comparing the solubilized intestinal disaccharide se complex * activity in the presence or absence of the inhibitor (so-called 100% value). As a substrate that determines the specificity of the inhibition sample, a practically low glucose-free sucrose (glucose 100 ppm)} enzyme activity is used based on the spectrophotometric determination of glucose released by glucose dehydrogenase and nicotinamide adenine dinucleotide as co. -Factor.

A saccharase inhibitor unit (SIE) is defined as the inhibitory effect that in a defined test condition Ο 5

DK 15 7 012 B

preparation reduces a given saccharolytic activity to one unit (saccharase unit = SE); The saccharase unit is defined herein as the enzyme activity which cleaves, under predetermined conditions, 1 µmol of sucrose per day. 1 minute and 5 thereby results in the release of 1 yamol of glucose determined by the sample and fructose not included in the sample.

The intestinal disaccharidase complex is recovered from porcine small intestinal mucosa by tryptic digestion, precipitation from 66% ethanol at -20 ° C, uptake of the precipitate into 100 mmol phosphate buffer with pH 7.0 and completed dialysis against the same buffer.

To 10 yal of a sample solution prepared so that the extinction of the sample preparation is at least 10% but not more than 25% below 100% value, add 100 15 µl of a dilution of the intestinal disaccharidase complex to 0.1 molar maleinate buffer, pH 6.25, and pre-incubate for 10 minutes at 37 ° C. The dilution of the disaccharidase complex is adjusted to an activity of 0.1 SE / ml.

Next, the saccharolytic reaction 20 is initiated by the addition of 100 µl of a 0.4 molar solution of

sucrose ("SERVA 35579") in 0.1 molar maleinate buffer, pH 6.25 and quenched for 20 minutes at 37 ° C by addition of 1 ml glucose dehydrogenase reagent (a small bottle of lyophilized glucose -dehydrogenase mutarotase mixture ("MERCK 14053") and 331.7 mg of O-nicotinamide adenine dinucleotide (free acid, "BOEHRINGER", purity Ί) dissolved in 250 ml of 0.5 molar tris buffer, pH

7.6). To detect the glucose, incubate for 30 minutes at 37 ° C and eventually photometer at 340 nm against blind reagent (with enzyme but without sucrose).

The calculation of the inhibitory effect of the inhibitor is made difficult by the fact that even very small changes in the test system, e.g. a 100% value that varies quite a bit from determination to determination has an impact on the 35 test result that you cannot ignore. You and

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DK 157012 B

6 goes through these difficulties, with a standard being followed by a standard; of a standard, a sucrose inhibitor of the formula C25H43 ° 18N8 has a specific inhibitory effect of 77,700 SIE / g and, when used, amounts of 20-30 ng in the sample lead to an inhibition of the order of magnitude above. Knowing the difference between the 100% value extinction and the extinction for the standard inhibited preparation, the extinction difference between the 100% value and the 10 of the test solution inhibited the preparation, taking into account the amount of inhibitor used. known way of calculating its specific inhibitory effect, expressed in saccharase inhibitor units per grams (SIE / g).

15 In vitro results of the sucrose inhibitor assay

Example No. Activity (SIE / g)

Desoxynoj irimycin (U.S. Patent No. 4,065,562) 466200 3 4662000

Example 1

Preparation of Starting Material 2,6-Imino-2-hydroxymethyl-2,6-didesoxy-L-ido (L-gulo) - hexonic acid nitrile

A solution of 46.6 g of 6-amino-6-deoxy-L-sorbo-furanose hydrochloride monohydrate is added to 200 ml of 0.5 N hydrochloric acid with 14.7 g of sodium cyanide and stirred for 3 hours.

Then, evaporate at 25 ° C in vacuo to a thin syrup, add 200 ml of methanol / ethanol (1: 1) and filter from the separated site. The filtrate is evaporated at 25 ° C in vacuo, the crystalline solid obtained is stirred with ethanol, suctioned off and washed with ethanol and diethyl ether. This gives 34.5 g (92% of theory) of color Ο

. DK 157012B

7 Loose crystals to a melting point of 156 ° C (dec.).

Rf value = 0.194 (floatant 1)

Rf value = 0.119 (floatant 2)

Rf value = 0.6 (flow agent 3) 5 Solvent 1 = chloroform / methanol / 25% ammonia in volume ratio 6: 4: 1 Solvent 2 = chloroform / ethyl acetate / methanol / 25% ammonia solution in volume ratio 40: 40: 30: 1 10 Solvent 3 = ethyl acetate / methanol / water / 25% ammonia solution in volume ratio 120: 70: 10: 2.

15

Example 2 2-Hydroxymethyl-3,4,5-trihydroxy-piperidine-2-carboxylic acid ethyl ester

OH

20. HO 0H

L JcC00CaH5

N OH

=: H

20 g of the compound of Example 1 is stirred for 3 hours at the boiling point in 240 ml of hydrogen chloride saturated ethanol and cooled and filtered off by suction. The filtrate is evaporated and the evaporation residue is purified on a 120 cm long and 6 cm wide cellulose column, the product being obtained as hydrochloride by elution with 90% acetone. The hydrochloride obtained is applied to an "Amberlite" ® IR 120-cat ion exchange column. Wash well with water and then elute with 0.5% ammonia. The fractions with the pure product are collected and evaporated. The evaporation residue slowly crystallizes under acetone by the addition of a little ethanol. 5.8 g of colorless crystals with m.p. 106-35 -108 ° C.

8

DK 157Q12B

ο

Example 3 2,2-Bis-hydroxymethyl-3,4,5-trihydroxypiperidine

OH

OH

Dissolve 1.2 g of the compound of Example 2 in 20 ml of absolute methanol and add 2 g of sodium borohydride in portions. Stir further for 2 hours, gently acidify with dilute hydrochloric acid and evaporate. The evaporation residue is dissolved in approx. 200 ml of methanol / water 1: 1 and applied to an "Amberlite" IR 12O cation exchange column.

Wash well with water. The product is obtained by elution with 2% ammonia. It is evaporated and the oily evaporator residue is dissolved hot in a little methanol and left to stand. After crystallization, it is filtered off by suction and washed with methanol. 0.6 g of colorless crystals with m.p. 148-150 ° C.

20 25 30 35

Claims (2)

Wherein R 2 is -COOH, -COOR 3 or -CH 2 -OH, and R 3 is a lower alkyl group, characterized in that 2,6-imino-2-hydroxymethyl-2,6-didesoxyphenyl; L-ido (L-guXo) -hexonic acid triX of the formX HO HO ”OH 15 o cn (III) è ch2oh-hydroxysated to a compound X with the formX 20 H ° vX> ° «Γ T ^ gooh," (V) JJ ^ "CH CE20E (VI)