SU1060118A3 - Process for preparing derivatives of glucoseamine - Google Patents

Abstract

Glucosamine derivatives of the formula <IMAGE> (X, R, R1, R2, R4, R6-R9 have the meaning stated in Claim 1) are prepared by condensation of corresponding 3-O-carboxymethylglucopyranosides with corresponding dipeptides. These glucosamine derivatives have the ability to increase the immunogenicity of an antigen on admixture therewith and, on systemic administration, to enhance the immunological reactivity of the treated organism. They can thus be used as adjuvants to increase the vaccine protection from bacterial, viral or parasitic pathogens.

Description

This invention relates to a process for the preparation of novel glucoamine derivatives with valuable pharmacological properties, in particular, a strongly pronounced immune enhancing effect. A known method for producing compounds expanding the arsenal of means of influencing a living organism by synthesizing the latter is based on the known cleavage reaction of the glucofuranose cycle under the influence of acidic reagents 1, the purpose of the invention is to obtain new glucosamine derivatives. This goal is achieved by the fact that according to the method of producing glucosamine derivatives of the general formula ... KO) - lol-g / 3iH-CO-Bi Lo-CH Kch 4 I t I 50-1GN-Sh-CO-Sh1-CH (JH212 " 5 .P) (L) (D) or phenyl; .-apkil where R is hydrogen or C.-C2-alky: T; hydrogen, C-C-alkyl or hydroxymethyl; R. - carboxyl, methoxycarbonyl carbamoyl, H- (C-C-alkyl) -carbamoyl, N-benzyl or n-carbamoylmethylcarbyl; RJ is carboxyl, methoxycarbonyl carbamoyl or N-methylcarb moyl, provided that the alkyl R contains more than 1 carbon atom, if R2 is methyl or hydrogen, a. and Rj is car-oxyl, a compound of the general formula ji Kch El IGG (JO-ISH-CH-CO-lSfH-CH- {(5H2) 2-B5 No.) (L) (pj where Ki-Kr- have the indicated values R-C-C-alkylidene or -cycloalkylidene is treated with acidic ion exchangers or trifluoroacetic acid for 10-25 hours at room temperature in 13de, tetrahydrofuran and / or methylene chloride. The target product is isolated with known methods. used such as those containing sulfonic acid groups, for example, Amberlite IR-120 (styrene resin with strongly acidic sulfo groups) or Dowex 50 (polystyrene sulfate new acids.) The starting materials used in the process can be obtained, for example, by introducing the acetamido-peptide residue R2 into the corresponding oxazoline with a free hydroxy group in position 3 of the sugar residue in one or several steps. The compounds of the invention have valuable pharmacological properties, in particular, clearly pronounced immunity enhancing effect 1. Strengthening cellular immunity in a living organism: an increase in the hypersensitivity of the late reaction type against ovalbumin in guinea pigs. Guinea pigs (according to Pittoright) on the first day of the start of the experiments were immunized with 10. mg of ovalbumin in Freund’s full auxiliary agent by injecting 5 t: 0.1 ml of the mixture consisting of the antigen and the auxiliary agent into each animal hind legs. 4 weeks later, the skin reacts by subcutaneous injection of 100 µg of ovalbumin in 0.1. W of a buffered saline solution and, based on the erythremic points calculated after 24 hours and the increase in the dermal swelling of the reaction volume, are quantified. The specific increase in antigen increase observed after 24 hours (late-type reaction), the reaction volume is a measure of the distribution of cellular and mimicity. Ovalbumin is too weak an immunogen to use as an acid or in an oil-water emulsion with an incomplete auxiliary agent (10 parts ovalbumin solution in 0.9% Iac1, mixed with 8.5 parts Bayol T and 1.5 parts ) to induce a late-type reaction. In order to effectively immunize it, it is necessary: but in the complete auxiliary agent, add to mycobacteria (5 mg of sacrificed and lyophilized M butyricum per 10 MP of Bayol F / araltsel A). In order to prove the immunity of the test substances, they can be added to the antigen-oil mixture instead of mycobacteria in: doses of 10-100 µg. Glucosamine peptides can imitate the action of mycobacteria and quantify them. The ED value of so-10 (µg / animal) in this example is 10, for example, in the following compounds X: 2-benzoyl. But-3-0- {1) -1 (D -1-carbamoyl-3-carboxypro IL) -l-carbamate J-carbamate -2-deoxy-1) -glucose, 2-benoylamino-3-0- {C1 -1- (D-1-carbamol-3-carboxypropyl) -1-carbamoylethyl-carbamoylmethyl - 2-deoxy D-glucose and 2-acetamido-3-0-l-1 (D-1-carbamoyl-3-carboxypropyl) carbamoyl ethyl carbamoylmethyl 2-deoxy-B-glucose.

A significant increase in late action ability against valbumin can also be achieved by not including this compound in the antigen-oil mixture, but administering it subcutaneously in doses of 10-100 µg each animal for several days after immunization (for example, B1, 2, 3 , 4, 5d.) In a solution of sodium chloride.

This shows that these compounds significantly enhance cellular immunity, both in a mixture with an antigen (an auxiliary effect in the narrow sense), and in their use independently of the time and place of injection of the antigen (systemic improvement of immunity).

2. Strengthening of humoral immunity in a living organism: an increase in the production of antibodies against bovine serum albumin (BSA) in mice.

NMRI-type mice are immunized by injecting 10 µg of BSA free from the precipitate intramuscularly on the first day of the experiment. After 9, 15 and 29 days. take a serum sample and examine it to determine the content of anti-BSA antibodies by passive haemaglutination. At used doses, soluble ECA is subimmunogenic for recipient animals, i.e. it can not cause the occurrence of antibodies or causes their occurrence in a minor amount. Additional treatment of mice with a specific, enhancing immunity, substance before or after antigen administration leads to an increase in antibody titers in serum. The effect of the treatment is expressed as the sum of the differences in the logj titers obtained during the three days of bleeding.

The compounds of the invention have an intraperitoneal or subcutaneous administration of 100-300 mg / kg per animal for five days after immunization with BSA to significantly increase the production of antibodies against BSA.

The effect of these compounds, which stimulates immunization, as opposed to other bacterial immunoleptics (for example, LPS from E. eoli) regardless of antigens: injections of new compounds show up

anti-BSA titer when applied only on immunized BSA mice. Subcutaneous administration of these compounds is as effective as intraperitoneal, i.e. observable

the effect of enhancing the imbalance of systems-. It does not depend on how the stimulant is administered in the same way as the antigen, or in a mixture with the antigen, as. when applying the known auxiliary agents.

Thus, these compounds can specifically develop (Hukural immunity, they improve the immunological response to

irritation, their enhancing immune action is based on the systemic activation of the immune apparatus. 3. Enhancing humoral immunity theta in vitro: replacement effect

T cells in the reaction of antibodies of the cells of the new spleen to sheep erythrocytes (CE).

To induce antibody response during

In many cases, lymphocytes (T cells) derived from the thymus are needed. These cells interact with the precursors of the antibody-forming lymphocytes 8 cells) and help them respond to stimulation by the so-called T-dependent antigens with proliferation, differentiation and antibody synthesis. Suspension of cells of Sesenka mice with congenital athymia

(type nu / nu) do not contain functional T-cells and cannot, for example, form anti-CE antibodies in a test tube in the presence of CE. These compounds can functionally

replace the T cells and make possible the antibody response against CE. The addition of these substances to the described cultures of spleen cells in the presence of CE results for

4 days to a significant increase

the number of cells that make antibodies. From the results of the experiments, it follows that these compounds increase the formation of humoral antibodies in the test tube and compensate for the defect of the T-cell system.

4. Selective mitogenicity for B-cells: causing a proliferation effect in B-lymphocyte culture.

for three days, incubate the suspension with a high content of B-lymphocytes (lymph node cells from mice with congenital athymia, such as nu / nu), as well as almost pure immature and mature T-lymphocytes (thymus cells or resistant to cortisone, i.e. after 48 h after cortisone injection, persistent thymus cells of Balb / c mice in the presence of test subjects

substances. A measure of proliferative activity is the incorporation of N-thymidine into lymphocytes during the last 18 hours of the culture period.

The compounds of the invention are mitogenic for B-lymphocytoB (i.e., for precursors of antibody-forming cells), but not for T-lymphocytes. They can stimulate the proliferation of lymphocytes, which are involved in the humoral immune response.

5. Compatibility.

Although these compounds have a potentiating effect on guinea pigs, for example, already after a single dose of 0.05 mg / kg subcutaneously, on a smaller subcutaneous dose after 5 times a dose of 10 mg / kg, no toxicity is observed after five times 300 mg / kg intraperitoneally. we Therefore, these substances have a wide therapeutic spectrum.

The compound of the invention has the property, on the one hand, in a mixture with an antigen to increase its immunogenicity, on the other hand, when systemically applied, to increase the immunological reactivity of the individual to his or her treatment of the organism. Moreover, these substances are able to induce cellular and humoral immunity and activate lymphocytes responsible for the formation of antibodies.

The novel compounds can thus be used as adjuvants in a mixture with vaccines in order to improve the effect of the vaccination and anti-infective agent based on humoral antibodies and / or cellular immunity against bacteria, viruses and parasites.

In addition, the proposed compounds in a mixture with a wide variety of antigens are suitable as auxiliary agents. In experimental and industrial preparation, the scientific research institute of antisera for therapy and diagnostics and in the induction of immunologically activated lymphocyte populations for transfer. cells.

The compounds can also be used without the simultaneous addition of antigen to promote an already started, but still invisible, immune response in a living organism. Therefore, the compounds are particularly suitable for stimulating the body's search, for example, in chronic. D1 or acute infections or in selective (antigen-specific) immunological defects that occur in old age or during severe primary diseases, and especially after treatment with ionizing rays or radiation. immune suppressing hormones. Thus, these substances can also be used, preferably in combination with anti-infective antibiotics, chemical-therapeutic agents and other methods of treatment, to counteract immunological damage to m.

P i m i r 1. Mix 2 g of 2-phenyl 4, (D-l5 -carbamoyl-3-carboxypropyl) -1-carbamoyl ethyl-carbamoylJ-5,6-0-isopropylidene-in-glucofurano) delta 2-oxazolin in water with 10 ml-Dowex-50-, H at room temperature for 15 hours. After filtering and drying, a colorless powder is obtained with a decomposition point of 140 ° C. The resulting 2-benzoylamino-3-0- {D-1-Ll- (D-1-car5 bamoyl-3-carboxypropyl) -1-carbamoylethyl-carbamoylethyl j-2-deoxy-f, iD-glucose contains the following conditions: Drying different amounts of water of crystallization. Yield 51%, 0.86 g.

Getting the raw materials. 3.77 g of 2-phenyl-4,5- |; s-O- (D-1-carboxyethyl) -5,6-0-isopropylidene-B-glucofuran-d-oxazolyl and 60 ml of acetonitrile and 15 ml of dimethyl5 formamide stirred with 1.4 ml of triethytamine and 2.55 g of N-ethyl-5-phenyl and zoxazolium-3 sulfonate for 1, 5 hours at 0 ° C, and almost all of them are dissolved. Then

0, 3.44 g of 1-amide- (b-alanyl-B-glutamic acid p-benzyl ester hydrochloride and another 1.4 ml of triethylamine are added and continued to stir for 24 hours.

5 at room temperature. It is evaporated under oil vacuum to a syrup and chromatographed on silica gel with a mixture of chloroform and acetone (8: 2). 6.5 g (97%) of a colorless 0 thick syrup are obtained.

2-phenyl-4,5-- 3-3-O-lL- (Dl-carb-amoyl-3-benzyloxycarbonylpropyl) -carbamoylethyl-l-carbamate-5, 6-isopropylidene-0-glucofur 5: but -L-oxazoline which

dissolving with the ester crystallizes. mp, 96-99 ° C,, (+ 13 ° (in chloroform),.

Crystalline 2-phenyl-4,5- {30-b-0-1-b- (B-1, -carbamoyl-3-benzyloxycarbonylpropyl) -carbamoylethyl-1-carbamoylethyl-5, 6-isopropylidene-O-glucofurano-l -oxazoline hydrogenates with 5% palladium on carbon in, dioxane at room temperature and normal pressure, and after evaporation in vacuo, 5.4 g (94%) of 2-phenyl-4,5-3-B-0-1 are obtained. -1, - (B-1 to Arb amoyl-3-to arboxypropyl) -carbamoylethyl-1-carbamoylethyl-5 .6-isopropylidene-B-glucofurano-D-oxazoline in the form of syrup. Calculated: C 51.40, H 6.26, N 9, 99 C24 H4 4-01-) 0.35 H 2 O (560.86) Found: C 51.13; H 6.38, N 9.89. Example 2. Starting material: 100 g of 2-phenyl-4,5- (5.6-0-isopropylidene-B-glucofurano) -D-oxazoline is dissolved in the absence of moisture and carbon dioxide in 1 l of acetonitrile and ijpH is added vigorous stirring at 15, .2 g of 55% dispersion of NaH - mineral y. oil and stir for 1 hour at room temperature. Then, 42 ml of ethyl chloroacetic acid ethyl ester are added dropwise with 42 ml of this mixture and after 1.5 hours another 42 ml. Through. 1.5 h add another 11.4 g of NaH-diPersia, mix for half an hour and add 42 m of chloroacetic ether dropwise at 0 ° C. After 2 hours, it is warmed to room temperature and evaporated in vacuo, and then in oil vacuum to a state of syrup, which is dissolved by ether, extracted three times by shaking with water, the ether phase is dried with sodium sulfate and after evaporation, 155 g of brownish are obtained. oil. This oil is dissolved in 150 ml of methanol and then a solution of 30 g of hydroxide is added to 150 ml of water, extracted two times with ether and the ether phase is washed once with water. The aqueous phases in vacuo are freed from ether and brought to pH 4 at a pH meter with 1N hydrochloric acid. ; The precipitated crystalline 2-phenyl-4,5-.3-0- (carboxymethyl) -5,6-0-from propyliden-r) -glucofyrano - l-oxazoline is sucked off, washed with water and dried. Obtain 107 g (93% of theoretical) with t. Pl. 186-. , (ot) |) -9 ° (CHCIj, с 0.9) and Cot) 2 ° (CHCIji с 3). 3.63 g of 2-phenyl-4,5- (3-0-carbox methyl-5, b-O-isopropylidene-B-glucofurano) -l-oxazoline, 3.43 g of hydrochloride compound 1-amide-y -benzyl ester of l-alanyl-P-glutamic acid, 1.21 g of m-oxysuccinic-, mida, 2.16 g of dicyclohexylcarbodimide and 1.45 ml of triethylamine are dissolved in 40 ml of dimethylformamide and stirred for 24 hours at room temperature temperature It is evaporated in an oil vacuum, dissolved with dichloro ethane and water, sucked away from the precipitated dipiclohexyl urea and extracted by shaking the organic phase twice with water and the aqueous phase twice with dichloroethane. After drying and evaporation of the organic phase, a thick syrup is obtained, which is chromatographed on silica gel in a mixture with chloroform - ethanol (9: 1). The resulting 2-phenyl-4,5- 3-0-Cl-L- (D-carbamone-3-benzyloxycarbonylpropyl) -carbamoylethyl-carbamate-5, 6-0-isopropylidene-B-glucofurano. D-oxazoline, which trituration with ether crystallizes, melts at 167168 ° C, (cC) | ° -5 ° (chloroform). 4.5 g of 2-phenyl-4,5- {3-0-1-L- (1-D-carbamoyl-3-benzyloxycarbonylpropyl) -carbamoyl ethyl-carbamoylmethyl-5, 6-0-isopropylidene-D-glucofurano - d2 α-oxazoline is hydrogenated in dirxane with 5% palladium on carbon, filtered from the catalyst and then eine is extracted with ethanol. The combined filtrates are evaporated and recrystallized from isopropyl alcohol. The resulting 2-phenyl-4,5- (-l-D-carbamoyl-3-carboxypropyl) -carbamoylethyl-J-carbamoyl-methyl-5, 6-0-isopropylidene-B-glucofurano-ω-oxazoline melts at 200-207 ° C. Obtaining the target product. 2.85 g of the resulting 2-phenyl-4,5- {3-O-1-B- (-lD-carbamoyl-3-carboxypropyl) -carbamoyl ethyl-carbamoylmethyl-5, 6-0-isopropylidene-b-glucofurano-I-4-oxazoline stirred in a mixture of 30 ml of water and 15 ml of tetrahydrofuran with 5 ml of Dowex-50-N for 15 hours at room temperature, filtered through a solid filter and evaporated to dryness in vacuo. By trituration with ether, a colorless powder of 2-benzoyl-amino-3-O- f is obtained | L-l- (D-1-carbamoyl-3-carboxypropyl) -1-carbamoylethyl-carbamoylmethyl -2-deoxy-, p-D-glucose with mp. 175-177 ° C (in the form of a hydrate). Yield 55%, 1.51 g. Calculated: C 51.11, H 5.97, N 10.37, C2HH (540.53) Found: C 50.95, H 6.04, N 10.23 Example 3 4.4g 2-4 enyl-4, 5- {3-O-C1-b- (1-P-3-dicarboxypropyl) -carbamoyl ethyl-carbamoylmethyl-5, 6-0-isopropylidene-B-glucofuraoxazoline is mixed for 20 hours at 40 ° C with 11 ml of Dowex-50 x 4 ion exchanger in a mixture of 45 ml of tetrahydrofuran and 20 ml of water. After filtration and cleaning with charcoal. (Darco G-60) solution is freeze-dried to obtain colorless amorphous 2-ben amido-2-deoxy-3-0-C1-L- {lD-3-di carboxypropyl) -carbamoylethylJ-car bamoylmethyl-C-glucopyranose with optical rotation (o.) ° + 25 ° (H ™ O, c 0.997). Yield 37%, 1.56 g. Preparation of starting material, b, 33 2-phenyl-4,5- (3-0-carboxymethyl-5, b-0-isopropylidene-C glucofurano L-oxazoline, 5.75 g 2 ethoxy-M-carbethoxy-1, 2-dihydroquinoline (EEDGl) and 9.3 ml of triethylamine were added to a solution of b-ala-nyl-in-glutamic acid dibenzyl ester trifluoroacetate (obtained from 8.3 g of N-dibenzene ester). Tper-butyloxycarb nyl-L-alanyl-Vgglutamic acid with 5.1 ml of trifluoroacetic acid and 2.6 ml of dichloroethane by four-hour hydrolysis at 40 s in 70 ml of dichloroethane. The mixture is kept for 15 hours at 40 ° C, p chloroform is added, extracted by shaking twice with water and the aqueous phases once with chloroform.After drying with sodium sulfate and evaporation and the chloroform solution contains 19.9 g of oil, which is purified with 400 g of silica gel / Merck by elution with ether, then chloroform - acetone 17: 3. At the same time, pure 2-phenyl-4,5- {s-0-1-b- (1-b-3-diben zyloxycarbonylpropyl) -carbamoyl ethyl-carbamoylmethyl-5, b-0-isopropyliden -P-glucofurano} - "z-oxazoline with m. Kip. FROM IB-C and U} 1 ° -47 ° (CHCI with 1.54). 7 g of 2-phenyl-4,5- {3-0-1-L-Y-0-3-dibenzyloxycarbonylpropyl) -cabamoylethyl -carbamoylmethyl-5, b-O-isopropylidene-U-glucofurano-J-L-oxazoline is hydrogenated with 1, 8 g of 5% Pd / C in a mixture of 80 ml of tetrahydrofuran and 20 ml. water until the reaction ceases, is sucked off from the catalyst, evaporated in vacuo and the residue is triturated with ether. This gives 4.9 g of 2-phenyl-4,5- {Z-O-lL- (1-B-3-; Sy1carboxypropyl) -carbamoylethylJ-carbamoylmethyl-5, 6-0-isopropylidene-0-glucofur. But J-y-oxazoline as a colorless powder. Calculated; C 48.13, H 6.08; N 7.32. .NaO ,, - 1.8 NdO (.5-73-, 9) Found: C, 48.59; -H, 6.07; N 7.52, Example 4. 2 g of 2-phenyl-4, (1-C-carbamoyl-3-tert-butoxycarbonylpropyl) -carbamoyl-2-hydroxyethylJ-carbamoylmethyl-5, 6-0-isopropylidene-O-glucofurano are hydrolyzed - d2-oxazoline for 20 h at room temperature with a mixture of 15 ml of methylated methylene and 5 ml of trifluoroacetic acid. It is evaporated in an oil vacuum, the residue is triturated with ether and 2-benzamido-2-deoxy-3-0-Cl-L- (lD-carbamoyl-3-carboxypropyl) -carbamoyl-2-hydroxyethyl 3-carbamoylmethyl-B-glucopyranose is obtained in the form of a beige powder, so pl. 100-115 ° C, (o) |, + 23 ° (H20, c 0.886), which is crystallized with 2 mol of water and 1 mol of trifluoroacetic acid. Rf 0.28; CHCgj: CHjOH 1: 1 (thin-layer silica gel. Merck). Output 60%, 1.16 g. The Radiation of raw materials. As in Example 3, 5.7 g of 2-phenyl-4,5- (3-0. -Carboxymethyl-5, b-0-isopropylidene-D-glucofurano) -d2 oxazoline is condensed with 4.9 g of o - hydrochloride. amide-j - L-seryl-D-glutamic acid tert-butyl ester with the addition of 2.3 ml of triethylamine and 5.2 g of 2-ethoxy-K-carbethoxy-1, 2-dihydroquinoline in 45 ml of dichloroethane. After 18 hours at 40 ° C crystallized forms. An additional 50 ml of dichloroethane is added, the mixture is cooled in an ice base, filtered, and the crystals are washed with cold diethlorethane. Analytically pure colorless crystals of 2-phenyl-4, 5- {3-0-1-L- (1-B-carbamoyl-3-tert-butyloxycarbonylpropyl) -carbamoyl-2-hydroxyethyl-carbamoyl-methyl-5, 6-0 -isopropylidene-B-glucofurano} - l2-oxazolin have so pl. 187-188 ° С (ot) 20 + 7 ° (СН, ОН, с 1,125). Calculated: C 42.60, H 5.27; N 7.99. B bj ° 12 2 f Оb HjO - О, 9 5 7 CF 3 COOH Found: C 42.87; H 5.32; N 8.20 Example 5. 6.4 g of 2-phenyl-4, 5- (lD- r -propylcarbamoyl-3-tert-butoxycarbonylpropyl) -carbamoyl ethyl-carbamoylmethyl-5, b-O-isopropylidene-B-glucofurano} - 4-oxazoline is hydrogenated with 1.8 g of S% -Horo Pd / C in 80 ml of tetrahydrofuran and 20 ml of water until the process is terminated, filtered from the catalyst and concentrated. Rf 0.58. SCHEZ: CHjOH 3: 1 (thin-layer silica gel. Merck), Yield 5.8 g. The resulting acid is then washed with 10 ml of the Dowex-50 X 4 ion exchanger, 50 ml of tetrahydrofuran and 25 ml of water for 15 hours at room temperature and 12 hours at 40 ° s After filtration, purification with charcoal (Darso-C-BO), secondary filtration and freeze-drying, a colorless, amorphous 2-benzamido-2.-Deso-xi-3-0-1-b- (1-DN-n-propylcarbamoyl-3 -carboxypropyl) -carbamoylethyl -carbamoylmethyl-D-glucopyranose, m.p. b5-140 ° C and ioL) + 28 ° (water, s 1.03), Rf 0.48 CHCEj: CHjOH 1: 1 (thin-layer silica gel, Merck), Yield 65% 4.15 g.

Source connection 5.25 g of 2-phenyl-4, .5- (3-0-carbosimethyl-5 f6-0-isopropylidene-B-glucofurano) -D-oxazoline and the trifluoroacetic acid salt of ot-p-propylamide- y-benzyl ester of L-ala nyl-O-glutamic acid,; - obtained from 6.2 g of oL - p-propyl-amide-f-benzyl ester of N-tert-butyloxycarbonyl-alanyl-B-glutamic acid and 4.2 ml of trifluoroacetic acid in 2.5 ml of dichloroethane, 6 hours at 40 ° C - in 60 ml of dichloroethane with the addition of 7.75 ml of triethylamine and 4.8 g of 2-etrxy-I-carbethoxy-1,2-dihydroquinoline. After 20 hours at 40 ° C, 50 ml of chloroform is added, shaken twice with water and twice with chloroform. After drying and concentrating the chloroform phase, 15 g of α-oil is obtained, which is purified on 200 g of sidicagel according to Merck by washing with ether, then with chloroform with acetone 7: 3. Get b, 4 g of colorless amorphous 2-phenyl-4,5-. - {H-O-lL- (lD-propyl-carbamoyl-3-tert-butoxycarbonylpropyl) -carbamoylethyl J-carbamoylmethyl-5, 6-0-isopropylidene-B-glucofurano - d2-oxazoline, Rf 0.35 СССЕ acetone 7 : 3 (thin-layer silica gel, Merck).

Calculated: C, 51.08; H 6,799, 16; HjO 4.72

N

2b zvCHO-11 1 2 ° (11И8)

H 7.00; Found: C 51.15;

4.77.

H 9.49,

"20

Example 6. 8.3 g of 2-phenyl-4, 5- {3-0-1-methyl-1- (lD-carbamoyl-3-tert-butyloxycarbonylpropyl) -carbamoyl ethyl -5.6-0-isopropylidene-O-glucofurano } - l-oxazoline is left to stand for 15 hours at room temperature in a mixture of 20 ml of trifluoroacetic acid, 60 ml of methylene chloride and 2 ml of water. It is then evaporated in vacuo and the residue is dissolved with ether. The resulting pink powder is dissolved in 200 ml of water and 0.5 coal (Da rco-G-60) is purified. After filtration and evaporation, a colorless amorphous 2-benzamido-2-deoxy-3-0-C1-methyl-1- ( D-1-carbamoyl-3-carboxypropyl) -carbamoyl ethyl-j-carbamoylmethyl-in-glucopyranose, m.p. 110-120 ° C, t) i) + 31 ° (HjO, c 0.88), Rf 0.52 acetone: ethanol

x: 1 (thin layer silica gel. Merck), which crystallizes with 0.6 mol trifluoroacetic acid and 1.7 mol water. Yield 52%, 4.15 g.

Getting the raw materials. 7.3 g of 2-phenyl-4,5- (3-O-carboxymethyl-5,6-O-isopropylidene-b-glucofurano) l-oxazoline, 6.5 hydrochloride d. -amide-j-T.-butyl ether

oi-amino-isobutyroyl-b-gluta new

0 Acid and 2.9 g of isobutyl chloroformate are dissolved in 25 ml of dimethylformamide and 50 ml of dichloroethane. To this is added dropwise at a temperature of from -15 to -10 ° C over 30 minutes to grow 6,1 ml of triethylamine in 20 ml of dichloroethane. It is then heated to room temperature and stirred for another 15 hours at room temperature. 50 MP of dichloroethane are added, shaken with water, twice with 0.5N. NaOH and three times with water, the aqueous phases .- twice with dichloroethane, the organic phase is dried and after evaporation 16.6 g of oil are obtained.

5 It is purified on silica gel / Merck, by removing the No. 1 van of CHCe.j: C. HdOH 19: 1, 9.7 g of colorless amorphous 2-phenyl-4,5-3-0-0-1-methyl-1- (lD -carbamoyl-3-tert-butyloxy0 carbonylpropyl) -carbamoyl ethyl 3-5,6-O-isopropylidene-B-glucofurano-oxazoline. Optical rotation (o (,) 5) + 6 ° (СНС1 ,, с 1.027),

m.p. 75-B9 ° C.

five

, 35, СНС1: С Н (thin-layer silica gel. Merck),

Calculated: From 46.31; H 5.86; N 8.57; HjO 4.69

  0.6 SRSC 1.7

0

VD-DJD: J

Found: C, 46.59; H 6.00; N 8.74, 4.53.

The following compounds are prepared in a similar manner.

4.1 g (85.7%) (B-l-cap5 basamyl-3-carboxypropyl) -carbamoyl-. ethyl carbamoylmethyl -2-ace.tamido-2-deoxy-B-glucose, (ot) 2o -10 ° + 1 ° (water, s 0.930).

Calculated: C 43.08; H 6.59;

0 N 11.16; Н20 4.67

.O -1.30 (501.87)

Found; C 42.85, H 6.61; N 11.26; Ho 4.67,

3.85 g (75.7% - :) 2-benzamido-25-deoxy-3-0-Li- (B-1,3-tert-N-methylcarbamoylpropyl) -carbamoyl ethyl-carbamoylmethyl-glucopyranose, t. square 125-132 ° С, (+ 24 ° (Н, О, с 0.93).

0

Calculated: C 50.55: H 6.89; N 11.91; HjO 5.51

 IS 50 (600)

Found: C, 50.18; H 6.81; N 11.65; HgO 5.73

five

2.43 g (50.4%) of 2-benzamido-2-deoxy-3-0-L-1- {D-1, 3-tert-methoxycarbonylgofopyl) -carbamoylmethyl-3-carbamoylmethyl-B-glucopyranose as a hydrate, t pl. 80-90 ° C., (Ot; i ° +25 (СНЗОП, с 1,017).

4.16 g (81%) of 2-benzamido-2-deoxy 3-0-Cb-1- (DlN-carbamoylmethyl-carbamoyl-3-carboxypropyl) -carbamoyl ethyl-3-carbamoyl-methyl-D-glucopyranose, t.p. 163-170S .

Calculated: C 49.02; H 6.34; N 10.99; H20 3.99

С2 (, Н,., И50 „1Н1 HjO (637.01)

Found: C, 49.85; H 6.48; Ы 10.64; EU 3.99

4.16 g (93%) 2-benz.amido-2-deoxy-3-0-1-1- (D-1-carbamoyl-3-carboxypropyl) -carbamoylpropyl-carb amoylmetyl-D-glucopyr anos a, t pl. 114-155 ° C.

Calculated: C 50.95; H 6.31) N 9.90; Ы20 1.98 С24 ЗА01Г ° b 2 (565.71)

Found: C 50.69; H 6.47, N 9.76; 1.98.

4.28 g (.85%) 2-benzamido-3-deoxy-3-0-1- (D-1-carbamoyl-3-carboxypropyl) -carbamoyl-2-methylproppropyl J - 1-carbamoylmethyl-D - glucopyranose, ((/) 2 ° + 32 ° (from 0.78, water).

Calculated: C 49,72; H 6.70; N 9.28, HjO 5.86

CorHj N Oi-t 15b% ° (603, .9)

Found: C 49.82; H 6.67} Ы 9.09; K20 5.86.

3.91 g (82%) of 2-acetamido-3-O- (D-1-carbamoyl-3-carboxypropyl) -carbamoylmethyl-carbamoylmethyl -2-deoxy-D-glucose, (oL) 2 ° +27 (c 0.944 , water).

Calculated: C, 43.97; H 6.08 N 12.06

 0, (464.43)

Found: C 43.72; H 5.88; N 12.20;

3.57 g (81%.) 2-acetamido-3-O- {y-1- (D-1, 3-dicarbamoylpropyl) -carbamoylethyl-carbamolmethyl} -2-deoxy-D-glucose, (S) 1 ° + 7 ° + 1 ° (water, with 0.514).

Calculated: C 43.26; H 6.78; N 14.01; NZO 4.46

C.gHj.N 50.6-1.24 H2 (499.81)

found: C 43.22; H 6.72; N 14.11; NZO 4.46.

4.54 g (95%; 2-acetamido-3- (1-1-carbamo-1-3-carboxypropyl) -carbamoylmethyl-1-carbamoyl-propyl J-2-deoxy-1) -glucose, - {s (./ || +46 + 1 (water, from 0.630) ...

Calculated: With 46.34 H 6.55; N 11.38

CigHN O ,, (492,48)

Found: C, 46.07; H 6.44; N.

4.17 g (87%) of 2-acetamido-3--0- (D-1, 3-dicarboxypropyl) -carbamoylethyl -carbamoylmethyl J-2-deoxy-D-glucose dimethyl ester, (c6da, c 0 , 0814).

Calculated: C 45.66 / H 6.75; 5 N 7.99 / HjO 3.54

С2оНз з0 2-.103 EUO (526.05). Found: C 45.90; H 6.70 N 8.00; H, 0 3.60

4.4 g (92%) 3-0- {Ll- (Dl-Kap10 bamoyl-1-3-carboxypropyl) -carbamoylethyl} -carbamoylmethyl -2-deoxy-2-p-propionamido B-glucose, (d) + 8 ° (water, with 1,146).

Calculated: C 42.90; H 6-, 93 5 N 10.53 H20 7.42

 , 19 NLO (531.93). Found: C 42.66 / H 7.01; G1 10.54; HPO 7.42

4.29 g C89%) 3-0- {Ll- (Dl-KapQ bamoyl-3-carboxypropyl) -carbamoylethyl-carbamoylmethyl} -2-kaprioylamido-2-deoxy-B-glucose a, (al-) + 11 ° (water, with 1.052).

Calculated: C 52.87, H 7.85 / 5 N 9, 4 9

C2e, H4bH40 ,, (590.67) Found: C .52.60; H 7.73; I-9.21,

4.55-g (95.2%) 2-acetamido-2-deQ hydroxy-3-0-UL-l- (Dl, 3-tert-methylcap50mylpropin) -carbamoylethyl -carbamoylmethyl JD-glucose, (dL + 31 ° (water, from 0.41) .-

 Calculated: C, 44.67 / H, 7.261 N, 13.02

С2оНз5К.; 0 о -179 2 ° (537.77) Found: С; G 7.40, N 13.31 .. -.

4.02 g (84%) of 2-acetamido-3-O- (L-1- (D-1-carbamoyl-3-carboxypropyl) 0 -carbamoyl-2-hydroxyethyl-carbamoylmethyl-Teale} -2-deoxy-P - glucose, (() 20 + 10 ° (water, about 1.653).

Calculated: C 43.72 / H 6.12

5 N 11.33

., (494.45) Found: C 43.51; H 6.03; N 11.11

4.41 g (92%) of 2-acetamido-3-OP (D-1-carbamoyl-3-carboxypropyl) -carbamoylbutyl-J-carbamoylmethyl -2-deoxy-1) -glucose a, (ot-) + 12 ° (water from 7.22) .-

Found: C 45.11; H 7.03; N 10.29; Н „0 4.38 С2оНзХо11-129 (529.75)

Included: C 45.35; H 6.99 N 10.58} TsuO 4.38

4.34 g (69.5%) (D-l-carbamal-3-carboxaxyl) -caro bamoylethyl-carbamoylmethyl -2-deoxy-2-stearoyl-amido-P-glucose, (oC) ° +24 (dimethylformamide, s

 0.663).

Calculated: C 58,10; H 8.89,

5 G 7.97; Cj., (702.89), Found: C, 57.92: H, 8.95; N 7.81. 4.55 g (95%) of 2-acetamido-3-O (B-1-car6amoyl-3-carboxyp-peel) -carbamoyl-2-methylpropyl-car moylmethyl} -2-deoxy-in-glucoea, (° C) | ° W of the Code, s 1,096). C, 45.53; H 6.98I N 10.62 H20 3.99 C.2o i PM40-, 1 1.17 HjO (527 Found: C 45.80; H 6.81, N 10.65; HrO 3.99. 4.41 g (92%) 2-acetamido-3-O-Cb-1- (D-1-carbamoyl-3-carboxypril drank) -carbamoyl-3-methylbutyl / - car bamoylmethyl-2-deoxy-D-glucose, U | 20 (water, p. 1.060). Calculated: C, 46.66; H, 7.16; N, 10.36; 3.71. Cl. Hj. NO -.- 1.11 HPO (540.53) Found: C, 46.45- , H 7.04 j N 10.06; EUO 3.71. 4.21 g (88%) 2-acetamido-3-O- {C-1- (D-carbamoyl-3-carboxypropyl) -carbamoylpropyl - carbamoylmethyl -2-deoxy-B-gl1ocose, (o (4-11 °; water, s 1,019). Calculated: C 44.27; H 6.79 N 10.87 H, 0 4.46 C., H, - 1.28 (515.54) Found: C 44.32; H 6.73; N 10.78; HjO 4.46. 4.13 g (86%) of 2-acetamido-3-O- b -1- (Dl-carbamoyl-methylcarbamoyl-3-carbox propyl) -carbamoylethyl J-carbamoylmethyl V2-deoxy-D-glucose a, white powder, (OH) IP (water, with 0.706). Calculated: C 43,24 H H 6,43, N 12,61; H20 3,61, - 1.11 (555.50) Found: SG43.10, H 6, 305N 12., 90; 3.61. 4.46 g (93%) of 2-trimethylacetamido2-deoxy-3, 0 - {Cb- 1- (D-1-carbamoyl 3-carboxypropyl) -1-carbamoylethyl carbamoylmethyl 1-ti, ft-D-glucoea, (d) 20-7 ° (c 0.6, methanol).

Claims (1)

(54) (57) METHOD FOR PRODUCING GL10COZAMINE DERIVATIVES of General Formula CH ₂ 0H J - o £ 0 \ -0H GCD — f / HH-C0-R1 T? 2 ~ CH I, V II ^s I * C 0 -UH-CH- C0-HH-SI- (SP3) 2 “ ^to 5 '(D) (L) (D) where R, is Cy-Cj-alkyl or phenyl; Yag is hydrogen or C, -C _e -alkyl; - hydrogen, C ^ -Cj-alkyl or hydroxymethyl; R ₄ is carboxyl, methoxycarbonyl, carbamoyl, N- (C ₁ -C ₃ алки -alkyl) carbamoyl, N-benzyl or N-carbamoylmethylcarbamoyl; R is carboxyl, methoxycarbonyl, carbamoyl or N-methylcarbamoyl, provided that alkyl R contains more than 1 carbon atom, if R _{2 is} methyl or hydrogen, and R. and R ₅ are carboxyl, characterized in that the compound is common formulas H ₂ ~ SP in _e in. СО-КН-СН-СО-ПН-СН- (СНз) ₂ -В 5 (D) W (D) where R ^ -Rj have the indicated meanings; ^R 6 ~ ^C 1 ~ C ₄ -alkylidene or ^C 5 ~ C ^ -cycloalkylidene, is treated with an acid ion exchanger or trifluoroacetic acid for 10-25 hours at room temperature in water, tetrahydrofuran and / or methylene chloride.