NL8702248A - PHOSPHORIZED ACYL GLYCOSIDES, THEIR PREPARATION AND USE. - Google Patents

Description

i k - Phosphorylated acylglycosides, their preparation and their use -

The present invention relates to phosphorylated acyl glycosides, methods of preparation thereof, pharmaceutical preparations containing them and their use as medicaments.

The invention particularly relates to compounds of the formula t, wherein represents a group of any one of formulas 2a-2k, wherein hydrogen represents a metal equivalent of 10 or alkyl, X and Y are the same or different and represent oxygen or an imino group and R 1 and R 8 are the same or different and represent an optionally substituted acyl group.

The compounds of the formula I can be prepared according to the invention by a) reacting a compound of the formula 3, wherein R 2> R ^ X and Y have the above meanings and R ^ is a protecting group, with a compound of the formula IV, wherein Rjr has the same meaning as Rj, but optionally hydroxyl groups which are present are in protected form and a group R ^ present in R ^ represents a group R ^ T, which is alkyl or a protecting group and Z is a represents a cleavage group to obtain a compound of the formula V in which R 4, R 4, R 4, X and Y have the above meanings and the introduction of a phosphate group into the compound of the formula 5, or b) reaction of a compound of formula 6, wherein R 2, R 1, R 1. , X and Y have the above meanings and Rg is a protecting group, with a compound of the formula IV to give a compound of the formula 24g-1-2mule 7, wherein, Rg, Rg Rg, R, X and Y have the above meanings, deprotection of the compounds obtained in protected form and recovery of the compound of formula 1 in free form or in acid addition salt form.

The reaction of a compound of the formula 3 or 6 with a compound of the formula 4 is carried out in an inert solvent, for example in a chlorinated hydrocarbon, such as dichloromethane, with the exclusion of moisture, preferably at room temperature. For the introduction of a phosphate group into a compound of the formula 5 according to method a), the compound of the formula 5 is dissolved in an inert solvent, for example in a chlorinated hydrocarbon, such as dichloromethane, or in a cyclic ether, at reduced or room temperature. such as tetrahydrofuran, and adds a protected phosphorus chloridate. The product can be recovered from the reaction mixture and, if necessary, purified in the usual manner. The cleavage of protecting groups can be carried out in one or more steps according to known methods. A protecting group can be removed, for example, by acid cleavage, for example with an aqueous acid (ion exchanger) or hydrogenolytically.

The protecting groups used are generally the protecting groups used in saccharide chemistry, for example, benzyl, alkyl, chloroacetyl, or optionally substituted silyl.

Known protecting groups can also be used as protecting groups for the phosphate radical, e.g. benzyl or phenyl.

R2 and Ri are the same or different and represent an acyl group with, for example, 4-20, preferably 12-16, especially 14 carbon atoms, which may be substituted in the 3 "position with OH, acetoxy or an acyloxy- group, where acyl is as defined above, wherein the C-3 atom is in the R or S configuration. Therefore, in the compounds of formula I, R 2 and R 2 may be present in achiral, R, S or racemic form. In the described reactions of the invention, the configuration remains unchanged in the course of the reaction, i.e. from R-, S- or racemic starting materials, the corresponding R-, S- or racemic end products.

The compounds of the formula I are preferably obtained as acid addition salts, wherein hydrophilic basic compounds are preferably used as counterions, for example to increase the solubility in water, for example tris (hydroxymethyl) aminomethane or L-lysine.

Z represents a leaving group, preferably halogen, especially chlorine or fluorine.

The starting materials of the formula IV are partly new and can be obtained by reacting a compound of the formula 4a, wherein the above meaning has 15 and Z 'represents acyloxy, in particular 4-nitrobenzoyloxy, or hydroxyl, with a hydrohalide or a Lewis acid .

Examples of such halides / acids are HCl, HBr,

Vilsmeyer chloride or bromide, thionyl chloride, TiCl 2, TiBr 2, or diethylamino sulfur trifluoride (DAST).

The starting materials of formulas 3 and 6 are known or can be prepared in analogy to known methods, for example as described in the examples.

The compounds of the formula I exhibit pharmacological action and are therefore suitable as drugs.

The compounds of the formula I exert a favorable influence in particular on the non-specific resistance to microorganisms in the following tests.

30 1. Determination of endotoxic activity in the limulus amoebocytes lysate test

Entotoxin catalyzes the action of a proenzyme in limulus amoebocytes lysate. The separation of p-nitroaniline from a colorless substrate is measured. The degree of secretion is measured photometrically, with the correlation between absorption and endotoxin concentration (or, endotoxin activity for analogs) being linear in the range 0.01-0.1 ng / ml endotoxin (comparison 1702 24 e $ * - 4 - with the absorption values of a standard toxin). They prepare a 1:10 dilution series from each sample (dissolved in pyrogen-free sterile, double-distilled water) .A blank sample is run with each determination series. 100 µl Sample, standard or blank sample are each treated with 100 µl limulus amoebocytes lysate. After incubation for 10 min at 37 ° C, 200 µl of substrate is added to the reaction. After an additional 3 min. Incubation, the reaction is stopped with 200 µl of 50% acetic acid.

After stirring the sample, the absorbance is measured in a spectrophotometer at 405 nm against a blank value. The endotoxin content (endotoxin activity) of the sample is determined as endotoxin units (E.U.) by calculating a linear regression with the values of the standard toxin.

15 2. Generation of endotoxin shock in the mouse

This trial observes the induction of an endotoxin shock or a clinically analogous condition with lethal outcome by LPS analogs in galactosamine (GalN) sensitized mice. Male C57 bl mice (6 animals per group) simultaneously receive i.p. 8 mg GalN, dissolved in 0.5 ml PBS and 0.1 µg LPS from Salmonella abortion equi (Sigma) dissolved in 0.2 ml physiological saline. This treatment results in the death of all animals within 6-12 hours. (C. Galanos et al., Proc. Natl. Acad. Sci., USA, 76_ (1979) 5939-5943). Instead of LPS in standard treatment, the test substances are administered once parenterally or orally in different doses, either simultaneously with GalN, or before or after treatment with GalN. Evaluation of the result is done by comparing the smallest doses, which are lethal for all animals in a group, or by calculating an LD ^ q using the Spearman-Karber method.

3. Microbial septicemia in the neutropenic mouse 35 -

This model allows for the testing of a substance-mediated increase in immunity in neutropenic mice infected with 87-5 2 24 8 i -5 microorganisms. For the induction of neutropenia, groups of 20 female B6D2Fi mice receive 1 x 200 mg / kg body weight s.c. cyclophosphamide, dissolved in 0.2 ml of distilled water, on day 0. On day 3, the test substance is administered parenterally (primary ip) or perorally, if possible dissolved in physiological sodium chloride solution, or otherwise dissolved in some way (0.3 ml). Infection is performed on day 4 by i.v. administration of the involved graft in a volume of 0.2 ml (number of germs 10 per mouse: eg Pseudomonas aeruginosa Δΐ2: 1x10 ~ *; E. coli Δ 120: 6 6 4 2x10; Staph, aureus Δ113: 1x10; Candida albicans Δ124: ίχ10).

The test animals are observed up to day 10 after infection and the deaths are recorded daily. The following parameters are evaluated by comparison with Iection Coitro animals 15 or standards: a) mean survival time b) survival rate

The compounds of formula 1 show remarkable improvements in survival time and moderate survival over untreated infected control animals in experimental infections with gram negative agents (eg.

Pseudomonas and E.coli) as well as infections with gram-positive agents (eg Staph, aureus) or yeasts (eg Candida albicans).

25 4. Activation of the oxidative metabolism of human blood neutrophils

Neutrophils (at least 95% pure) are incubated with the test substance in three different concentrations for 1-2 hours at 37 ° C. The release of the cells by superoxide anions as an activation index to the reduction of cytochrome C inhibable by superoxide dismutase is then measured. 1x6 neutrophils, whether or not pretreated with the test substance, are added to cytochrome C (80 µMol), which formylmethionylleucylphenyl -35 alanine (10 µM). The controls additionally receive 50 µg of superoxide dismutase. After incubation for 15 min at 37 ° C, the reaction is stopped by cooling the test tubes in an ice bath. The test tubes are then centrifuged at 400 x g for 5 min to remove the cells. The reduction of cytochrome C, which is proportional to the amount of superoxide anion released, is measured photometrically at 550 nm. The inhibitory effect of the test substance 5 on PMN activation by LPS is measured as described, except that after incubation of the leocyte cells, incubation is continued with a stimulating concentration of LPS.

5. Carbon Disappearance Test 10 This test is based on the principle that particles of size 200-250 Å (eg carbon particles) can be removed from the organism by phagocytosis by macrophages only. After intravenous administration, carbon particles are removed from the bloodstream by macro-phage phagocytosis in the liver (Kupfferstellate cells) and the spleen.

The RES activation of a substance is tested by single or repeated administration in solution in water or as a suspension. The amount of substance intended for treatment is injected i.p. or s.c. administered in 4 single daily doses, or 20 at once, 24 hours before the start of the trial. A suspension containing 10% carbon black is diluted with a 1% gelatinous sodium chloride solution to a level of 16 mg cabbage / ml. Each mouse receives 0.2 ml / 20 g body weight i.v. 25 μΐ of blood is collected by puncture of the orbital plexus after 3, 6, 9, 12, 25 and 15 min after i.v. administration. The blood is hemolyzed in 2 ml of distilled water. The carbon concentration is then determined photometrically. The animals are then sacrificed and the weight of the livers and spleens is determined.

30 6. Herpes infection (mouse)

The intracutaneous herpes infection model allows the testing of a substance-mediated increase in the immunity of mice infected with herpes viruses. The course of the disease is monitored and allows the observation of the successive appearance of different parameters. Herpetic lesions appear at the site of infection, then ulcerate, the neighboring leg becomes paralyzed, and the paralysis continues to increase until death. Immunocompetent "hairless" (hairless) mice are infected intracutaneously on day 0 by intracutaneous administration of the involved graft in a volume of 0.025 ml (e.g. 1x10 plate-forming units Herpes simples type Ι / mouse). The substance to be tested is injected i.p. administered in solution, for example, in physiological saline (0.1 ml).

The systemic herpes infection model allows the testing of a substance-mediated increase in the immunity of mice infected with herpes viruses. Immunocompetent NMRI mice are infected on day 0 by Ί5 i.p. administration of the concerned graft in a volume of 0.1 ml (e.g., 1.3x10 "* plate-forming units of Herpes simples type 1 / miis).

The substance to be tested is s.c. administered in solution, for example in physiological saline.

The test animals are observed up to day 20 after infection and the occurrence of injuries, paralysis and death is recorded. The following parameters are measured and compared with infection control animals or a standard: a) number of mice with lesion (cumulative) b) number of mice with paralysis (cumulative 25 c) average survival time d) survival rate.

In the experimental HSV-1 infection, the compounds of the compounds of the formula I cause marked improvements in the course of the disease, the survival time and the survival rate compared to untreated control animals. These effects are observed after a single i.p. or s.c. administration between days 0 or -1 and +6.

7. Colony Stimulating Factor (CSE) Generation 35 CSFs are mediators produced by the organism due to infections or the effects of toxins. Their biological functions are complex and they are observed through stimulation of the proliferation of the hemopoietic 870 2 24 8 i.

- 8 - system, in particular of bone marrow cells. B, D0F, mice receive the test substances once or repeatedly, parenterally or orally up to a dose of 50 mg / kg. Serum from the test animals is collected at different times. The CSF-5 ....

activity titers of the sera are measured in a cell culture assay based on the proliferation rate of bone marrow cells of B2D2F1 (iz / j). Metcalf, The Hemopoietic Solony Stimulation Factors, 1984, Elsevier; T. Mosmann, J. Immunol. Method, 65 (1983) 55-63; L.M. Green et al., J. Immunol. Methods, 70 (1984) 257-268, /.

10

The compounds of formula 1 elicit CSFs in various sizes in mice at doses from about 0.025 to about 25 mg / kg. Time-kinetic difference in CSF activities is also observed, which could be beneficial in therapeutic use.

15 8. Generation of interleukin-1 (IL-1)

The determination of whether substances stimulate cells to produce IL-1 is primarily performed in a cell culture assay. First, both resident and thioglycolate-licensed adherent macrophages are recovered from mice.

These are incubated in RPMI medium for 48 hours with different concentrations of test substance and the liquids above the cell are collected. These are assayed for IL-1 activity in cultures of thymocytes from CHH / HeJ mice. The thymocytes of these supernatants containing IL-1 produced by macrophages are incubated for 72 hours to proliferate. The proliferation is measured by incorporation of 3 H-thymidine in a scintillation counter (J. Gery et al., J. Exp. Med. 136 (1972) 128-141; J. Oppenheim et al., Cellular Immunol. 50 (1980) 71 -81).

30 -; -; -

The compounds of the formula I exhibit varying degrees of IL-1 production induction.

in macrophages at concentrations from about 0.1 to about 50 µg / ml.

9. Generation of LPS (endotoxin) tolerance (lethality tolerance)

Parenteral administration of LPS one to three times daily in the mouse can induce a so-called tolerance that protects the animals from the lethal effects of LPS after administration of galactosamine (compare endotoxin shock induction with the mouse above).

The compounds of the formula I already provoke such a tolerance at a single i.p. with different administration of 0.25 mg / mouse. Pretreated (tolerant) mice are exposed to an LPS challenge at a dose of 0.01 µg LPS + 8 mg galactosamine / mouse i.p. at different times (1 day to 3 weeks) after the last treatment.

10

A majority of animals survive this LPS challenge, especially after repeated dosing (3 times) compared to untreated challenged control animals.

Furthermore, the compounds of the formula I have anti-inflammatory effect, in particular at 15 ....

specific inflammation induced by immunologically induced and hypersensitivity agents and allergic reactions. This activity can be demonstrated by various test methods, for example by investigating the influence on prostaglandin synthesis in vitro and in vivo. In vitro, the inhibition of LPS and zymosan-induced PGE2 and PGF10C release has been investigated. Thio-glycolate-stimulated peritoneal leucocytes from NMRI mice are incubated with LPS or the test substance for 24 hours. After a change of medium and washing three times, the cells are stimulated for 2 hours with LPS or 1 hour with zymosan.

PGE2 and PGF2 are observed in the supernatants.

Inhibition of both LPS and zymosan-stimulated PGE2 production of the cells is found.

In vivo, inhibition of by LPS is measured

induced PGE9 release by mouse peritoneal leucocytes after pre-30. z handling with substances to be tested. Groups of 3 NMRI mice are turned up 1,2 and 3 i.p. treated with LPS, or substance to be tested, respectively. On day 4, these animals and a control group received 1.5 ml of thioglycolate i.p. and on day 7, peritoneal mouse leucocytes are collected and stimulated with LPS. A marked reduction in PGE2 release is found compared to control animals.

In a further test, the influence on procoagulant action (PCA) is investigated. After stimulation with LPS, human endothelial cells synthesize PCA as measured by a shortening in plasma coagulation time. The coagulation time of re-calcified plasma is also shortened by mouse peritoneal macrophages obtained after LPS treatment and by rabbit peritoneal leucocytes after induction of the generalized Shwartzman reaction. Stimulated overnight with LPS alone or with LPS and test substance in DMEM medium without fetal calf serum (FCS), trial design a) J, respectively. In experimental design b), mouse peritoneal leucocytes are treated with LPS or test substance for 24 hours and stimulated with LPS overnight after medium change. The coagulation time of re-calcified human control plasma is measured using the Hakchen method after adding the mouse peritoneal leucocyte suspension, which has been repeatedly frozen and ultrasonically treated.

Addition of the test substance reduces the LPS-induced PCA compared to the control value, as evidenced by an increase in coagulation time. Pretreatment with test substance also results in reduction of PCA.

hi vivo, the influence of LPS-induced PCA in mouse peritoneal leucocytes after pretreatment of the animals with test substance can be demonstrated as follows: B-D-F. mice are injected i.p. on days 1, 2 and 3 treated with LPS 25 6 2 or test substance respectively. On day 3, all animals additionally receive 1.5 ml thioglycolate i.p. On day 6, peritoneal leucocytes are collected, the sample from each animal is adjusted to 1x10 cells / ml and stimulated with LPS in DMEM medium without FCS for 24 hours. The PCA of these cells is as above 30 ....

described determined. Pretreatment with LPS or test substance, respectively, produces a markedly reduced PCA. A similar reduction can be observed in rabbits. The PCA of rabbit peritoneal leucocytes can be reduced after induction of the generalized Shwartzman reaction and by pre-treatment with LPS or test substance, respectively.

For the study of the inhibition of the local Shwartzman reaction, groups of three chincilla rabbits 870 2 24 8-11 - i.v. or i.p. pretreated with LPS or test substance on days 1, 2 and 3. On day 6, LPS is administered intradermally in 40, 20, 10, 5, 2.5 and 1.25 µg per skin site. On day 7 the reaction is started with 2 jig LPS / kg i.v. The experiment is evaluated semi-5 quantitatively by examination of the skin necroses. Almost complete inhibition of the Shwartzman reaction is observed after pretreatment with LPS or test substance, respectively.

Furthermore, the compounds of formula 1 exhibit anti-tumor activity, as demonstrated in the following experiments: 1. Tumor necrosis factor (TNF) induction

To stimulate mouse bone marrow maerophages, bone marrow cells from BDF mice (ca. 10-13 days old) are raised

r I

with CSF) diluted to 1x10 cells / ml in medium / RPMI 1640 + 1% Pen / Strep (5000 U / ml) + 1% glutamine J and in flat microplates with the test substances in solution in the range of 100 jig to 0 , 1 µg / ml final concentration (dilution steps 1:10) incubated for 4 hours at 37 ° C / 5% (X ^. After filtration through 0.45 µm filters, the supernatants are frozen at -70 ° C until needed .

L 929 cells were cultured overnight (37 ° C, 5% CX 4) in microtiter plates at 3 x 10 4 cells / well / 100 µl, then added 100 µl of each sample and further diluting the culture in 1: 2 steps. 100 µl actinomycin D (8 µg / ml medium) is added and incubation is continued for another 18 hours at 37 ° C / 5% (X ^.

After removal of the supernatant, the remaining cell layer is stained with Giemsa solution and the absorbance is measured at 620 nm with a Titertek Multiscan Auroreader (flow). One unit of TNF is defined as the activity, which results in 50% lysis of the intended L 929 cells.

2. B16F1 melanoma test

B16F1 melanoma cells are grown in vitro for 5 days. The cells are synchronized 1 day prior to the assay by shredding the monolayer into individual cells using EDTA trypsin and returning the whole amount to the same fresh medium vial. The cells are counted and placed on 10 cells / ml medium. 0.1 ml of cell suspension (ΙΟ "* cells) is injected intravenously into the tail vein of B, DF mice. 21 days later, the mice are sacrificed and the number of lung tumors counted. The substance to be tested is dissolved and injected intraperitoneally on days -6, -4 and -1.

The compounds of formula 1 exhibit immunoprophylactic activity, which is reflected in a reduction in the number of B16F1 melanoma metastases in the lungs. As a test of therapeutic effect, mice are treated on days 3, 6, 8, 10, 13 and 15 after inoculation of the tumor cells. Here, too, a marked reduction in the number of metastases is observed.

The compounds of formula I are therefore suitable as modulators of nonspecific resistance to microorganisms in the systemic enhancement of immune response and in enhancement of non-specific immunity. Thus, the compounds of formula 1 are suitable for the treatment or supportive treatment (ie together with other specific or supportive therapeutic forms) of conditions accompanied by decreased immune response, in particular decreased humoral immune response and / or reduced hypersensitivity reaction. delayed type and in the treatment of conditions, generally requiring modulation of immune response. The compounds of formula 1 are particularly suitable for the treatment or supportive treatment of pathological conditions based on ideopathic immunological deficiencies or immunological deficiencies of the type occurring in geriatric patients 50, or in patients with severe burns or genera initiated infections. The compounds of the formula are also suitable for the treatment or supportive treatment of viral diseases (such as disseminated Herpes, progressive smallpox and disseminated varicella diseases), of Morbus Hodgkin 35 and other malignant tumors. Furthermore, the compounds of the formula I are suitable for the prevention of endotoxin shock, for example in accidents, burns and surgical interventions and 870 2 24 8 i-13 - as anti-inflammatory agents.

For the above indications, the dose naturally varies depending on the compound used, the host, the mode of administration and the nature and severity of the condition to be treated. In general, however, it is indicated that satisfactory results in animals are obtained at a daily dose, or in a single administration to obtain an adjuvant effect, for example, in supportive treatment, from about 0.0015 mg / kg to about 1 mg / kg 10 body weight of the animal. In larger animals, for example humans, the indicated daily dose is from about 0.1 mg to about 70 mg of the compound of the formula 1, which, in the case of a daily dose, can be given well four times a day. An indicated total single adjuvant dose is up to 70 mg of the compounds.

In view of their immunomodulatory activity, the compounds of the formula I are also suitable as excipients in vaccines. For this use, an indicated dose is about 0.5 mg to about 100 mg, preferably about 70 mg, administered on the day of vaccination, with a repeat of the same dose being appropriate 2-4 weeks later.

The compounds of the formula I can be administered by any convenient route, generally enterally, preferably orally, for example in the form of tablets or capsules, or parenterally, for example in the form of injectable solutions or suspensions.

The compounds of formula 1 can be administered in free form or in the form of a pharmaceutically acceptable acid addition salt. Such salts can be prepared in a conventional manner and exhibit activity in the same order as the free bases. The present invention also relates to pharmaceutical compositions containing a compound of the formula 1 in the form of the free base or in the form of a pharmaceutically acceptable acid addition salt, in association with at least one pharmaceutical carrier or diluent. Such preparations can be prepared in the usual manner.

870 2 24 8 - 14-

Dosage unit forms contain, for example, about 0.025 mg to about 35 mg of the compound of the formula 1 in free base form or in the form of a pharmaceutically acceptable acid addition salt.

The invention also relates to a method of combating diseases and infections as described above by administering to a subject in need of such treatment a therapeutically effective amount of a compound of the formula 1 or 3 or, where applicable, a pharmaceutical acceptable salt thereof. The invention also relates to a compound of the formula 1 or 3 for use as a medicament, in particular as an immunomodulator, as an anti-virus and as an anti-inflammatory agent.

The following examples illustrate the invention. Temperatures are given in ° C.

15

Example I; 6-0- (0C-D-Arabinofuranosyl) -2-deoxy-4-0-phosphono-3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamid (7-D-glucopyranose).

Di-tris salt (method a) a) Benzyl-6-0- (2,3,5-tri-O-benzyl-^ -D-arabinofuranosyl) -2-deoxy-3-0-tetradecanoyl-2- /3-(R)- tetradecanoyl- oxytetradecanamidq? - ft-D-glucopyranoside 25 270 mg Benzyl-2-deoxy-3-0-tetradecaonyl- S '- 2 - /. 3- (R) -tetradecanoyloxytetradecanamido / - pD -glucopyranoside are well pre-dried and dissolved in 50 ml of dry dichloromethane (distilled over Ρ90ς). 3 g of powdered Drierite (preheated in high vacuum to 160 [deg.]) And 2 g of silver carbonate are added. A solution of 2,3,5-tri-O-benzyl-1-chloro-1-deoxy-ö (-D-arabinofuranose (freshly prepared from 600 mg of 2,3,5-tri-) is added dropwise with vigorous stirring. 0-Benzyl-1-0-4-nitrobenzoyl-0 (, 5-D-arabinofuranose) in 50 ml dry dichloromethane Completely anhydrous conditions must be maintained The reaction is completed after about 2 hours (DC control: R ^ = 0.54 C in toluene / ethyl acetate = 2 / 1J) The solid of the clear solution is filtered, the solvent is evaporated and the residue is separated into two series-connected 870 2 24 8-15. Size B Merck-Lobar silica gel columns; linear gradient eluent of toluene / ethyl acetate (7/1), followed by toluene / ethyl acetate (3/1) each 800 ml.

5 b) Benzyl-6-0- (2,3,5-trl-0-benzyl-fr-arabinofuranosyl) -2-deoxy-4-Q-diphenylphosphono-3-0-tetradecanoyl-2- / 3- (R ) -tetradecanoyloxytetradecanamidq7-j-D-glucopyranoside: 1 g Benzyl-6-0- (2,3,5-tri-0-benzyl-o-D-arabinofuranosyl) -2-deoxy-3-0-tetradecanoyl-2 - / 3- (R) -tetradeeanoyl-10-oxytetradecanamido / p-D-glucopyranoside is dissolved in benzene and evaporated several times and then taken up in 30 ml of freshly distilled (P 2 O) dichloromethane. 278 mg of 4-dimethylaminopyridine (evaporated 3 times with dried benzene) and 1 ml of freshly distilled dry pyridine are added. Thereafter, 810 ml of diphenylphosphoric chloridate are introduced directly into the solution under an argon atmosphere. DC control: (toluene / ethyl acetate = 5/1) shows virtually no residual starting material (Rf = 0.20) after 3-4 hours. For work-up, the reaction mixture is shaken with potassium hydrogen sulfate, sodium bicarbonate and finally water, and the organic phase is dried over magnesium sulfate and concentrated by evaporation. The residue is chromatographed on a silica gel column (Merck Lobar Type C); eluent: toluene / ethyl acetate = 5/1, R4 = 0.4.

C) 6-0- (<X, -D-Arabinofuranosyl) -2-deoxy-4-0-diphenylphosphono-3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyl-oxytetradecamido ^ -D-glucopyranose : 100 mg Benzyl-6-0- (2,3,5-tri-0-benzyl-D-arabinofuranosyl) -2-deoxy-4-0-diphenylphosphono-3-0-tetradecanoyl-50 2- / 3 "(R)" tetradecanoyloxytetradecanamidoj-tb-D-glucopyranoside is suspended in 15 ml absolute ethanol and hydrogenated at room temperature and atmospheric pressure for 8 hours in the presence of palladium on active carbon (10%). The starting material slowly dissolves during this process. The catalyst is finally filtered over celite and the solvent is evaporated. Mp. 87-92 ° (from methanol). R ^ = 0.46 (in ethyl acetate).

870 2 24 8 * - 16 - d) 6-0- (C 1 -D-Arabinofuranosyl) -2-deoxy-4-O-phosphono-3-O-tetradecanoy1-2- / 3- (R) -tetradecanoyloxytetradecanamino / -D-glucopyranose (Di-tris salt): 30 mg 6-0- (fcl-D-Arabinofuranosyl) -2-deoxy-5 4-0-diphenylphosphono-3-0-tetradecanoyl-2- / 3- (R tetradecanoyloxy tetradecanamido-D-glucopyranose is taken up in 5 ml glacial acetic acid.

15 mg of platinum oxide is added and hydrogenated at room temperature and atmospheric pressure for 2 hours. Thereafter, the band indicating presence of starting material in DC (R ^ C ^ .0.93) is no longer visible and a new band appears at R ^ W 0.55 (in chloroform / methanol / glacial acetic acid / water = 5/16/1/1).

Example II; 2-Desoxy-6-Q-0 (-D-glucopyranosy1-4-0-15 phosphono-3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamido7-D-glucopyranose (process a)) a) Benzyl -6-0- (2,3,4,6-tetra-O-benzyl-D-glucopyranosyl) -2-deoxy-3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyl-oxytetradecanamido7 ~ ft-D-glucopyranoside; 1 g of Benzyl-2-deoxy-3-O-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamido7-t5-D-glucopyranoside are taken up in 50 ml of dichloromethane, which is freshly distilled. (È) is about 1.5 and 3 g of powdered Drierite (dried under high vacuum at 160 DEG C. for 8 hours) and 6 g of silver carbonate are added.

The glycosyl halide obtained from 3.07 g of 2,3,4,6-tetra-O-benzy-1-O-1-4-nitrobenzoyl-D-glucopyranose is slowly added to the reaction mixture (ca 50 ml) under exclusion of light. After 1 hour, 3 g of silver carbonate is added and the reaction mixture is reacted for a total of about 24. The solids are then filtered off, the filtrate is concentrated by evaporation and chromatographed on a 50 x 6 cm medium pressure column (gradient: toluene / ethyl acetate = 7/1 and toluene / ethyl acetate = 3/1, 800 ml each). The title compound is eluted as a mixed fraction together with 2,3,4,6-tetra-O-benzyl glucose. The separation is performed on a Merck Lobar silica gel column, type C (conditioned with hexane / ethyl acetate = 3/1) using two successive gradient elutions: 870 2 24 8-17; 1) hexane / ethyl acetate = 3/1 and hexane / ethyl acetate = 2/1 each, 800 ml.

2) hexane / ethyl acetate = 2/1 and hexane / ethyl acetate = 1/1, each 800 ml.

2 b) Benzyl-6-0- (2,3,4,6-tetra, -0-benzyl-K - D-glucopyranosyl) -2-deoxy-4-0-diphenylphosphono-3-0-tetradecanoyl-2 - / 3- (R) -tetradecanoyloxytetradecan amide / -ff-D-glucopyranoslde:

Hen dissolves 700 mg of benzyl-6-0- (2,3,4,6-tetra-0-benzyl-θί-D-glucopyranosyl) -2-deoxy ~ 3-0-tetra-1 q decanoy1-2- / 3 - (R) -tetradecanoyloxytetradecanamido / - ff-D-glucopyranoside in 20 ml of dichloromethane, fresh from distillation.

184 mg of 4-dimethylaminopyridine and 1 ml of freshly distilled pyridine are added. This is brought directly into a solution of 517 ml of diphenylphosphoric acid chloride under argon atmosphere and the mixture is stirred at room temperature for 72 hours. The reaction is not complete.

DC: 0.39 (in toluene / ethyl acetate = 5/1). To work up, 1 ml of water is added to the mixture, stirred for 10 min and the result is shaken with sodium hydrogen sulfate and sodium bicarbonate.

The organic phase is dried over magnesium sulfate, the solvent-2Q evaporated and the residue chromatographed on a type C Merck Lobar silica gel bowl (1200 ml toluene / ethyl acetate = 6/1; then 800 ml toluene / ethyl acetate = 3/1).

c) 2-Desoxy-4-0-diphenylphosphono-6-0-P (.- D- glucopyranosyl-3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradeanamido7-D-glucopyranose:

150 mg of benzyl-6-0- (2,3,4,6-tetra-0-benzyl-c * -D-glucopyranosyl) -2-deoxy-4-0-diphenylphosphono-3-0-tetradecanoyl-2 are suspended - / '3- (R) -Tetradecanoyloxytetradecanamido J-β-Ό-30 glucopyranoside in 40 ml 96% ethanol and add 33 mg 10% palladium on activated carbon. It is hydrogenated at room temperature and atmospheric pressure for about 7 hours. The catalyst is filtered off over celite and the filtrate is concentrated by evaporation. The title compound is obtained as a colorless syrup. R ^ ftO ^ S (in chloroform / 35 £ methane / glacial acetic acid / water = 16/5/1/1).

d) 2-Desoxy-6-O-D-glucopyranosyl-4-O-phosphono-3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamidci7-D- 870 2 248 è-18 - glucopyranose:

80 mg of 2-deoxy-4-0-diphenylphosphono- 6-0-¾ -D-glucopyranosyl-3-0-tetradecanoyl-1-2- / 3- (R) -tetradecanoyl-oxytetradecanamido-D-glucopyranose are dissolved in 25 ml glacial acetic acid. 40 ml of platinum oxide are added and hydrogenation is carried out at room temperature and atmospheric pressure for about 80 minutes. The catalyst is then filtered off, the glacial acetic acid is removed by evaporation with toluene 3 times, the residue is taken up in 40 ml of 96% ethanol and 14.7 mg of tris (hydroxymethyl) aminomethane (= tris) are added. When nan nan 50 ml of toluene are added, everything dissolves. The solution is concentrated by evaporation and then evaporated twice on a rotary evaporator with a small amount of absolute ethanol.

Rj = 0.63 (chloroformin thanol / glacial acetic acid / water = 5/16/1/1).

15

Example III: 2-Desoxy-6-O-methyl (3-deoxy-ft-D-manno-2-octulopyranosyl) -ono-4-0-phosphono-3-0-tetradecanoyl-1-2- / 3 (R) -tetra-decanoyloxytetradecanamido / -, ^, -D-glucopyranose (process a)): 20 a) Benzyl-6-O- / methyl (tetra-0-chloroacetyl-3-deoxy-,, ft-D-manno -2-octulopyranosyl) ono7-2-deoxy-4,0-diphenylphosphono-3-O-tetradecanoyl-2-.Z3- (R) -tetradecanoyloxy-tetradecanamido / -V * -D-glucopyranoside

98 mg of 4,5,7,8-tetra-0-chloro-25-acetyl-2,3-dideoxy-2-fluoro-1-D-manno-2-octulopyranosonic acid methyl ester and 100 mg of benzyl-2-deoxy are dissolved -4-0-diphenylphosphono-3-0-tetra-decanoyl-1-2-3- (R) -tetradecanoyloxytetradecanamidq7-f-D-glucopyranoside in 200 ml of dry dichloromethane. 200 mg of a 4A powdered molecular sieve and, after cooling to 0 °, about 3 drops of borane trifluoride etherate (BF2 * OEII) are added. The mixture is then stirred at 0 ° for 30 minutes and at room temperature for an additional 24 hours (thin layer control: dichloromethane / ether = 7/1). The mixture is then poured onto ice-cold sodium bicarbonate aqueous solution and extracted with dichloromethane and the organic phase is dried over anhydrous sodium sulfate and concentrated by evaporation.

The residue is chromatographed on silica gel (a: dichloromethane, b: dichloromethane / methanol = 500/1). The compound is first obtained as the fastest eluting component, followed by the compound, m.p. 870 2 24 8 - 19. 47-50 °.

M-Compound - 9.4 ° (c = 0.36 in chloroform) ^ Compound = 4.0 ° (c = 0.55 in chloroform).

b) Benzyl-6-O- / methyl (3-deoxy-^ -D-manno-2-octulopyranosyl) onQ7-2-deoxy-4-0-diphenylphosphono-3-Q-tetradecanoyl-2 / 3- ( R) -Tradecanoyloxytetradecanamid "f> -D-glucopyranoside

30 mg of benzyl-6-Q- / methyl (tetra-O-chloroacetyl-3-deoxy-oi, -D-manno-2-octulopyranosyl) are dissolved ono7-2-deoxy-4-0-diphenylphosphono-3-0 -tetradecanoyl-2- / 3- (R) -tetradecanoyl-oxytetradecanamido / - (- - D-glucopyranoside in 0.6 ml glacial acetic acid and add 1.2 ml 2,6-lutidine. The reaction mixture is cooled 12 to 0 And adding 0.48 ml of hydrazine dithiocarbonate with stirring, the mixture is then stirred at 0 ° for 2 hours (thin layer control: dichloromethane / methane = 10/1) and then concentrated together with toluene at a bath temperature of 30-35 ° C. The residue is extracted with dichloromethane and the solution is washed cold with dilute hydrochloric acid, water, dilute soda solution and ice water, dried, and concentrated by evaporation. The residue is chromatographed on a silica gel column (a: dichloromethane; b: dichloromethane / methanol = 100 /). 1, c: dichloromethane / methanol = 50/1) The title compound is thus obtained.

Mp. = 54-56 °, ~ 1.7 ° (c = 0.24 in chloroform), c) Benzyl-2-deoxy-6-0- / methyl (3-deoxy-ft-D-manno-2-octulopyranosyl ) -ono7 ~ 4-0-diphenylphosphono-3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamida7-fo-D-2q glue opyrano side

Mai treats a mixture of 51 mg benzyl-6-0- / methyl (3-deoxy-Xi-D-manno-2-octulopyranosyl) ono ^ 2-deoxy-4-0-diphenylphosphono-3-0-tetradecanoyl-2- 3- (R) -tetradecanoyloxytetra-decanamid-7-D-glucopyranoside, 1 ml glacial acetic acid and 2 ml 2.6-32 lutidine with 0.8 ml hydrazine dithiocarbonate solution as described under b).

Mp. 62-65 °, 4.3 ° (c = 0.35 in chloroform).

870 2 24 8 - 20 - d) 2-Desoxy-6-0- / methyl (3-deoxy-ft-D-manno-2-octulopyranosyl) onQ7-4-0-diphenyl-osfono-3-0-tetradecanoyl- 2- / 3- (R) -tetradecanoyloxytetradecanamido7 ~ ^, AD-glucopyranose

Benzyl-6-O- / methyl (3-5 deoxy-ft-D-manno-2-octulopyranosyl) -on-7-desoxy-4-O-diphenylphosphono-3-O-tetradecanoyl-2- / 3- ( R) -tetradecanoyloxytetradecanamido7-T5 -D-glucopyranoside in ethanol / methanol over palladium activated carbon.

Mp = 58-60 °, E & J-q ~ 22 ° (c = 0.31 in chloroform) .- 10 e) 2-Desoxy-6-0- / methyl (3-deoxy-fi-D-manno-2 -octulopyranosyl) ono / -4-0-phosphono-3-0-tetradecanoy1-2- / 3- (R) -tetradecanoyloxytetradecanamido7-p (, ft-D-glucopyranose 6-0- / Methyl (3-deoxy-ft - D-raanno-2-octulopyranosyi ono) -2-deoxy-4-0-diphenylphosphono-3-0-tetradecanoyl-2-r / 3- (R) -15 tetradecanoyloxytetradecanamidoZ-1H, ft-D-glucopyranose is dissolved in ethanol and hydrogenated over platinum oxide (Adam's catalyst) to give the title compound.

Mp. 138-139 ° C, LWJ-q = 26 ° (c = 0.26 in ethanol / chloroform = 3/1).

Example IV

2-Desoxy-6-jA-0- (3-deoxy-D ^ -D-manno-2-octulopyranosyl) -acid methyl ester-3,5-dideoxy-0 (-D-arabino-2-octulopyranosylic acid methyl ester) 7-4-0 ~ phosphono-3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamido2-D-glucopyranose (process b)): 25 a) Benzyl-2-deoxy-6-0- / 4-0- (3- deoxy-4,5: 7,8-diisopropylidene-^ -D-manno-2-octulopyranosyl) -acid methyl ester-3,5-dideoxy-7,8-isopropylidene-D-arabino-2-octulopyranosylic acid methyl ester 7-4-0- diphenylphosphono-3-O-tetradecanoyl-2 - /, 3- (R) -tetradecanoyl-30-oxytetradecanamidqT-ft-D-glucopyranoside:

A solution of 146 mg of /3,5 -didoxy-7,8-0-isopropylidene -4-0-(3-desoxy-4,5:7,8-di-0-isopropylidene-0 -manno-2-octulopyranosyl) -oic acid methyl ester - ^ - D-manno-2-octulo-pyranosyl / onacid methyl ester in absolute dichloromethane, react with 35 64 mg of chloromethylenedimethyliminium chloride (Vilsmeier reagent) and stir for 2 hours at room temperature. shaken saturated sodium bicarbonate solution, dried over magnesium 870 2 24 8 e - 21 - sulfate and concentrated by evaporation The residue is dissolved in 5 ml of absolute dichloromethane and added dropwise to a suspension of 574 mg of 2-deoxy-4-0. -phosphono- 3-0-tetradecanoy1-2- / 3- (R) -tetradecanoyloxytetradecanamido / -D- ^ glucopyranose, 76 mg Hg (CN) 2, 36 mg HgBr2 and 0.5 g molecular sieve 4A in 5 ml absolute dichloromethane After stirring at room temperature for 48 hours, the reaction mixture is filtered and after concentrated by evaporation, the residue is chromatographed with tolu one / ethyl acetate (2/1) over silica gel. The title compound is obtained as a colorless syrup.

VNMR: 7.45-7.09 (m, 15H, arom); 5.73 (d, 1H, 0-9.5, N-H);

JMci9Z

5.24 (dd, 1H, J2 3 ^ 11.0, J3 4 ^ 9.0, H-3); 5.03 (dddd, 1H, CH-OR), 4.92 and 4.62 (J ^ U ^ CgH ^ O); 4.59 (d, 1H, J1 ^ U8.5, H-1); 4.51 (ddd, 1H, 13 J ^ n git 8,0, J4 "3tie'v4,5, J4" 2 »3λγ3,0, Η-4"); 4,47 (dd, 1H, J4 9, 5, H-4); 4.43-4.36 (m, 2H, H-4 *, 7 "); 4.29 (dd, 1H, J5 "6lA2,0, H-5"); 4.14 (dd, 1H, J8 '* a, 81¾ ^ 9' J8 ", 7" ^ * 10> H-8 " a) ί M7 (dd, 1H, J8 "b? I1 ^ 5,0, H-8" b); 4,08-3,67 (m, 7H); 3,63 (dd, 1H, H-6 "); 3.59 (s, 3H, CH3OCO); 3.56 (s, 3H, CH30CÖ); 2.95 (dd, 1H, J3" e 3 "& η / 15.0, H-3" e) 2.43 (dd, 1H, 20 J3 * e 41 ^ 5.0 ^ -3 ^ ^ 2.36 ^, 96 (m, 6H, CH2-CO); 1.90 (dddd, 1H, H-5'e); 1.09 (dd, 1H-, H-3f, a); 1.73-1.00 (m, 82H, H-3, a, 5'a, (0¾), (CH3) 2C), · 0 .87 (t, 12H, CH 3).

b) 2-Desoxy-6-0- / 4 ~ 0- (3-deoxy-fe4-D-manno-2-octulopyranosyl) -acid methyl ester-3,5-dideoxy-C (-D-arabino-2- 25 -1 - octulopyranosylonic acid methyl ester 7-4-0-phosphono-3-0-tetradecanoyl-2-13- (R) -tetradecanoyloxytetradecanamido ^ -D-glucopyranose:

The sequential removal of the benzyl and phenyl protecting groups of benzyl-2-deoxy-6-0- / 4-0- (3-deoxy-4,5: 7,8-diisopropylidene-c / -D-manno-2- octulopyranosyl) 30-acid methyl ester-3,5-dideoxy-7,8-isopropylidene-D-arabmo-2-octulo-pyranosylic acid methyl ester / 4-0-diphenylphosphono-3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamido / PD-glucopyranoside is carried out in analogy to Examples I-III. The acid-catalyzed hydrolysis of the isopropylidene group takes place at the same time and the resulting title compound has + 3.20 ° (c = 0.2 in chloroform / methanol = 2/1).

870 2 24 8 - 22 -

Example V

2-Desoxy-6-0- £ 5-0- (3-deoxy- ¢ (-D-manno-2- * octulopyranosyl) onacid methyl ester 7-0 (-D-arabinofuranosyl ^ -4-0 ----- phosphono- 3-0-tetradecanoyl-2 - / [3- (R) -tetradecanoyloxytetradecanamidQ // - D-glucopyranose: 5 a) Benzyl-2-deoxy-6-6- / 2,3-di-0-benzyl- 5- 0 ~ Z (3-desoxy-4,5: 7,8-diisopropylidene - ((D-manno-2-octulopyranosyl) -oonic acid methyl ester 7-Q <-D-arabinofuranosyl7-4-0-diphenylphono-3-0- tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamido7 -D-10 glucopyranoside:

Hen dissolves 280 mg of 2,3-di-0-benzyl-5-0-Z (3-deoxy-4,5: 7,8-diisopropylidene-0 (-D-manno-2-octulopyranosyl) acid methyl ester J-D -arabinofuranose (dried in high vacuum over calcium sulfate) or in 10 ml of dichloromethane. Add 120 mg of chloro-15 methylenedimethyliminium chloride (reagent from Vilsmeier) to a suspension of 0.5 g of molecular sieve 4A, (in high vacuum over calcium sulfate at 180 Dried in a mixture of 25 ml of dried dichloromethane and 2 ml of dried acetic nitrile and stir the reaction mixture for 10 min with the exclusion of moisture.

This suspension is added to a freshly prepared solution of the starting material in dichloromethane and stirring is continued for a further 10 minutes.

The reaction of the reducing disaccharide derivative to halide is followed by reacting the respective thin layer chromatography samples with methanol / silver carbonate to methyl glycoside on an analytical scale. Once the formation of the disaccharide halide is complete, the reaction mixture is poured into cold saturated sodium hydrogen carbonate solution and extracted immediately with dichloromethane. The organic phase is concentrated on magnesium sulfate by evaporation and the resulting syrup is dried on calcium sulfate under high vacuum. 250 mg of mercury cyanide, 80 CF mg of mercury bromide, 2 g of molecular sieve 4A and 1.7 g of benzyl-2-deoxy-4-0- (diphenyl) phosphono-3-0-tetra-decanoyl-2- / 3- (R) are suspended -Tetradecanoyloxytetradecanamido-7-D-gluco-pyranoside, all powdered and dried in high vacuum over calcium sulfate, in 40 ml of dried benzene. 20 ml of benzene are distilled off. After cooling, a solution of the freshly prepared disaccharide chloride in 25 ml of dry benzene / dichloromethane (1/1) is added after cooling and the reaction mixture is stirred for 18 hours with the exclusion of moisture. The solids are filtered over celite, the filtrate is concentrated by evaporation and the products are separated by column chromatography over silica gel (gradient: toluene / ethyl acetate = 5/1/2/1). In toluene / ethyl acetate (2/1) the 0t-ara product has an R 1 = 0.44; = + 12.4 ° (c = 0.44 in chloroform); the £ -ara product: = 0.36.

^ b) Deoxy-6-0- / 51-0 - / (3-deoxy-o (-D-manno-2-octulopyranosyl) -acid methyl ester J- ° (-D-arabinofuranosyl7-4-0-phosphono-3-0- tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamida / -D-glucopyranose: 15.

Successive removal of the benzyl and phenyl protecting groups of benzyl-2-deoxy-6-0- / 5,3-di-0-benzyl-5-0 - / (3-deoxy-4,5: 7,8- diisopropylidene-CD-manno-2-octulopyranosyl) -acid methyl ester at 7 ° C. -D-arabinofuranosyl / -4-O-diphenylphosphono-3-O-tetradecanoyl-2- / 3- (R) -tetradecanoyloxy-tetradecanamido / -W-D-glucopyranoside is carried out in analogy to Examples I-III. The acid catalyzed hydrolysis of the isopropylidene groups occurs simultaneously. The resulting title compound has BÜ-q + 1.4 ° (c = 0.5 in dichloromethane / methanol = t / 1).

25

Example VI: 2-Desoxy-6-O-3-desoxy - ((D-manno-2-octulo-pyranosyl) -acid 7-2- / 3- (R) -hydroxytetradecanamido / ~ 4-0-fo spono - 3- / 3- (R) -tetradecanoyloxytetradecanoyl / -D-glucose: 30 a) Benzy 1-6-0- / f (3-deoxy-4,5,7,8-tetra-0-chloroacetate-0 ( -D-manno-2-octulopyranosyl) -acid benzyl ester / - 2- / 3- (R) - (benzyloxymethoxy) tetradecanamido / -2-deoxy-4-0-diphenylphosphono-3-0- / 3- (R) - tetradecanoyloxytetradecanoy-D-glucopyranis ide: 35

A mixture of 0.13 g of benzyl-2- / 3- (R) -3- (benzyloxymethoxy) -tetradecanamid-7-deoxy-4-0-diphenylphosphono-3-0- / 3- (R) - is stirred tetradecanoyloxytetradecanoyl2- ^ -D-glucopyranoside (M. Kiso et al., Carbohydrate Research, 162, 247-256 /. 1987, 8702248 4 - 24 - 1 ml dry dichloromethane, 0.1 g molecular sieve 4A, 55 mg mercury cyanide and 19 mg of bromide of mercury at room temperature for 3 hours under exclusion of moisture A solution of 0.1 g (3-deoxy-4,5,7,8-tetra-0-chloroacetyl-D-manno-2-octulopyranosyl-bromide) is added Acid benzyl ester in dry dichloromethane and stir the reaction mixture at room temperature for 4 hours After completion of the reaction (thin layer control: dichloromethane / ether = 7/1), the solids are filtered off and washed with dichloromethane IQ. Filtrate and wash solutions are combined with sodium bicarbonate solution shaken, dried and concentrated by evaporation The residue is chromatographed on silica gel (dichloromethane and dichloromethane) / methanol = 500/1); - 20 Z & 7d = + 12 ° (c = 1.8 in dichloromethane).

B) Benzyl-6-0- / T (3-deoxy-O ^ -D-manno-2-octulopyranosyl) -acid benzyl ester-2- / ¾- (R) - (benzyloxymethoxy) -tetradecanamidq7 ~ 2-deoxy -4-0-diphenylphosphono-3-0-Z3- (R) -tetra-decanoyloxytetradecanoyl / -D-glucopyranoside: 2q With a solution of 30 mg benzyl-6-0- / '(3-deoxy-4, 5,7,8-tetra-0-chloroacetyl- (-D-manno-2-octulopyranosyl) -oic acid benzyl ester 7-2 (3) (R) - (benzyloxymethoxy) tetradecanamidq7_2-deoxy-4-diphenylphosphono-3 ~ 0- / 3- (R) -tetradecanoyloxytetradecanoyl7-Φ-D-glucopyranoside in 1.8 ml 2,6-lutidine / acetic acid (2/1) 25, 0.48 ml freshly prepared hydrazine dithiocarbonate is added at 0 °.

The mixture is stirred at 0 ° for 2 hours and then evaporated under vacuum at 35 ° bath temperature. The residue is taken up in chloroform and washed successively with 1M hydrochloric acid, water, aqueous sodium bicarbonate and water. The solution is dried and concentrated by evaporation. The residue is chromatographed on silica gel (a: dichloromethane; b: dichloromethane / methanol = 100/1; 20 c: dichloromethane / methanol = 50/1); J & J-q = + 8 ° (c - 1.0 in dichloromethane).

C) 2-Desoxy-6-0- / 3-deoxy - ^ - D-manno-2-octulopyranosyl7oic acid-4-0-diphenylphosphono-2- / 3- (R) -hydroxytetradecanamido-3-0- £ ¾- (R) -tetradecanoyl · oxytetradecanoyl · 7 ~ D ~ glucose:

78 mg of benzyl-6-O-3-deoxy-8 7 0 2 2 4 8 - 25 - are hydrogenated

Or -D-manno-2-octulopyranosyl) -acid benzyl ester 7-2- / 3- (R) - (benzyl-oxymethoxy) tetradecanamidq7 ”2-deoxy-4-0-diphenylphosphono-3-0- ^ 3- (R) -tetradecanoyloxytetradecanoyl7 D-glucopyranoside in 20 ml of ethanol / methanol (3/2) in the presence of 80 mg of 10% palladium 5 on activated carbon at 40 °. After completion of the reaction (thin layer control: dichloromethane / methane = 10/1 and 2/1), the catalyst is filtered off and washed with ethanol / methanol / dichloromethane.

The filtrate and wash solutions are combined and concentrated by evaporation. The title compound is obtained. Mp.

138-140 °; SO = + 23 ° (c = 0.6 in ethanol / dichloromethane = 3/1).

d) 2-Desoxy-6-O- /% 3-deoxy-o (-D-manno-2-octulopyranosyl) -acid / -2-ƒ3- (R) -hydroxytetradecanamido7-4-Qf osf ono-jg- ( R) -tetradecanoyloxytetradecanoyl J-D-glucose: 15 In a solution of 35 mg of 2-deoxy-6-0- (3-deoxy-0 (-D-manno-2-octulopyranosyl) -acid J-4-O-diphenylphosphono-2 -3- (R) -hydroxytetradecanamido-3-0-ƒ3- (R) -tetradecanoyloxytetra-decanoy17-D-glucose in 15 ml of ethanol 30 mg of pre-reduced platinum catalyst of Adam are added and the mixture is stirred at room temperature. from the reaction, the catalyst is filtered over celite and washed with ethanol / methanol / dichloromethane. Filtrate and washing solutions are combined and concentrated by evaporation. After cleaning by preparative thin layer chromatography (dichloromethane / methanol = 2/1), the title compound is obtained.

Mp. 162-164 °. & 7β = + 25 ° (c = 0.5 in dichloromethane / ethanol = 1/1).

Example VII: 2-Desoxy-6-O- / -3-deoxy-1 * -D-manno-2-octulopyranosyl) acid 2-4-0-phosphono-3-0-tetradecanoy1-2-ƒ3- (R) -tetradecanoyloxytetradecanamido ^ -D-glucose: a) Benzyl-6-0 - / (3-deoxy-4,5,7,8-tetra-0-35 chloroacetyl - ^ - D-manno-2-octulopyranosyl) -acid benzyl ester / - 2-desoxy-: 4-0-diphenylphosphon ono-3-0-tetradecanoy 1-2-./3-(R)- tetradecanoy 1-oxytetradecanamidoV-fi-D-glucopyranoside: 8702248 - 26 -

In analogy to example V1) obtained from benzyl-2-deoxy-4-O-diphenylphosphono-3-O-tetradecanoyl-2 -3- (R) -tetradecanoyloxytetradecanamidq7 - $ -g-glucopyranoside and (3-deoxy-4,5 7,8-tetra-O-chloroacetyl-D-manno-2-octulopyranosyl-carbonone bromide) acid benzyl ester. Mp. 32-33 °. = + 10 ° (c = 1.3 in dichloromethane).

b) Benzyl-6-0- / ~ (3-deoxy-fr (-D-manno-2-octulopyranosyl) -acid benzyl ester J-2-deoxy-4-0-diphenyl-10-phosphono-3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamido7 ~ fe-D-glucopyranoside:

In analogy with example VIb.

Mp. 56-58 °. / 5 <7ρ = + 2 ° (c = 1.6 in dichloromethane).

C) 2-Desoxy-6-0- / 7 (3-deoxy-h (-D-manno-2-octulopyranosyl) -acid J-4-0-diphenylphosphono-3-0-tetradecanoyl-2- / 3- ( R) -tetradecanoyloxytetradecanamido7 ~ D ~ glucose:

In analogy with example Ic).

20 M.p. 114-115 °. = + 13 ° (c = 1.3 in ethanol / dichloromethane = 3/1).

d) 2-Desoxy-6-0 - ^ (3-deoxy- ^ -D-manno-2-octulopyranosyl) -acid 7 "4-0-phosphono-3-O-tetradecanoy1-2- / 3- (R) -25 tetradecanoyloxytetradecanamido7 ~ D ~ glucose:

In analogy to example 1d).

Mp. 168-169 °. = + 22 ° (c = 1 in methanol / dichloromethane = 1/1).

Example VIII

2-Desoxy-6-Q- [3-deoxy-P (-D-manno-2-octulopyranosyl) -acid 7-4-0-phosphono-2- / 3- (R) -tetradecanoyloxytetra-decanamido7-3-0 -./3-(R)- tetradecanoyloxytetradecanoyl7-D-glucose: oca) Benzyl-6-0 - / '(3-deoxy-4,5,7,8-tetra-0-chloroacetyl- <KD-manno-2 -octulopyranosyl) -acid benzyl ester 7 ~ 2-deoxy-4-0-diphenylphosphono-2- / 3- (R) -tetradecanoyloxytetradecanamidQ? ~ 3- 0-./3-(R)"tetradecanoyloxytetradecanoylV- β-D-glucopyranoside: 87 2 4® - 27 -

In analogy to example V1) obtained from benzyl-2-deoxy-4-0-diphenylphosphono-2- / 3- (R) -tetradecanoyloxy-tetradecanamido2 “3-0- ^ 3- (R) -tetradecanoyloxytetradecanoy1 / -ψ-D glucopyranoside and (3-deoxy-4,5,7,8-tetra-O-chloroacetyl-D-5 manno-2-octulopyranosyl bromide) acid benzyl ester.

= + 12 ° (c = 1.8 in dichloromethane).

b) Benzy 1-6-0- / j (3-deoxy-o (~ D ~ manno-2-octulopyranosyl) -acid benzyl ester / -2-deoxy-4-0-diphenylphosphono- 2- / 3- (R) - tetradecanoyloxytetradecanamidqy-3-0- / 3- (R) -tetradecanoyl-oxytetradecanoyI7 ~ P1 -D-glucopyranoside:

In analogy to example VIb).

[OiJ-Q = 48 ° (c = 1 in dichloromethane).

c) 2-Desoxy-6-0 - / - (3-deoxy-fi -D-manno-2-octulopyranosyl) impure -4-4-diphenylphosphono-2- / 3- (R) -tetradecanoyloxy-tetradecanamido7-3 -0- / 3- (R) -tetradecanoyloxytetradecanoyl-D-glucose:

In analogy with example Ic)

Mp. 138-140 °. & L = + 23 ° (c = 0.6 in 20 D ethanol / dichloromethane = 3/1).

* d) 2-Pesoxy-6-Q- / (3-deoxy-D (-D-manno-2-octulopyranosyl) -acid 7 ~ 4-0-phosphono-2- / 3 ~ (R) ~ tetradecanoyloxytetradecanamido7- 3-0- / 3- (R) -tetradecanoyloxytetradecanoyl / -D-glucose: 25 -

In analogy to example 1d).

Mp. 162-164 °. = + 26 ° (c = 0.5 in ethanol / dichloromethane = 1/1).

The compounds 30 required as starting materials can be obtained in the following manner.

A) Benzyl-2-deoxy-3-Q-tetradecanoyl-2-Z3- (R) -tetradecanoyloxytetradecanamidq7- D-D-glucopyranoside (for example I) 35 and dissolves 500 mg benzyl-2-deoxy-4,6 0- isopropylidene-3 ~ 0-tetradecanoyl-2 - / [(3- (R) -tetradecanoyloxytetradecana-mido / -fD-glucopyranoside) in 80 ml freshly distilled tetrahydro- 87 0 2 µg> - 28 - furan. add a mixture of 2 ml 40% hydrofluoric acid and 4 ml tetrahydrofuran in portions at room temperature and with vigorous stirring The reaction is followed by thin layer chromatography and is completed after 24 hours R ^ 0.28 (in toluene / ethyl acetate = 5 l / 1).

B) Benzyl-2-deoxy-3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamido ^ -ft-D-glucopyranoside (for example 1) Ίq 2 g of benzyl-2-deoxy-4,6 are dissolved - O-isopropylidene-3-O-tetradecanoyl-1-2- / 3- (R) -tetradecanoyloxytetradecanamidoj - - - - - D-glucopyranoside at 20 ° C in 90 ml of trifluoroacetic acid at ice bath temperature and maintain this temperature for approx. 7 min. .

The reaction is then almost complete.

DC control: R ^ (^ 0.28 (in toluene / ethyl acetate = 1/1). Most of the trifluoroacetic acid is evaporated and the residue is partitioned between dichloromethane and sodium bicarbonate solution. The organic phases are dried over magnesium sulfate and evaporation concentrated and the residue is chromatographed on a silica gel column (gradient: toluene / ethyl acetate = 2/1 and pure ethyl acetate, 400 ml each).

6) 2,3,5-Tri-Q-benzyl-1-chloro-1-deoxy-d (-D-arabinofuranose (for example I): 600 mg of 2,3,5-tri-O-benzyl are dissolved - 1-0-4-nitrobenzoyl-c (5-D-arabinofuranose in 50 ml dry dichloromethane and pass dry HCl gas until no further precipitation of 4-nitrobenzoic acid occurs. Thin reaction chromatography is carried out to check that the reaction is complete The precipitated 4-nitrobenzoic acid is then filtered off, washed quickly with dried dichloromethane, and the dichloromethane solution is quickly shaken with sodium bicarbonate, after which it is concentrated on magnesium sulfate and used directly for glycosylation.

The degree of hydrolysis of the glycosyl halide can be determined as follows:

A DC sample is added to a drop 870 2 24 8

»P

* - 29 - Peel dried methanol and add a trace of silver carbonate.

This mixture is shaken several times for 5 min and placed on a DC plate. The methyl glycoside is quantitatively formed from the halide originally present: R-0.37 (in toluene / 5% ethyl acetate = 10/1). The 1- * 0H compound remains unchanged (R ^ 0.11).

D) 2,3,4,6-Tetra-O-benzy1-1-chloro-1-deoxy- (X-D-glucopyranose (for Example II)

Hen dissolves 2,3,4,6-tetra-O-benzyl-1-0-4-10.

nitrobenzoyl-D-glucopyranose xn analogy to example C in dichloromethane and pass dry HCl gas (for 2 hours at ice bath temperature and then stand for 24 hours).

DC: (nitrobenzoate) = 0.44; R ^ (chloride) 0.5 (in toluene / ethyl acetate = 10/1). After quantitative reaction, the 4-nitrobenzoic acid is filtered off and the filtrate is concentrated on a rotary evaporator to remove most of the HCl gas, after which it is quickly shaken with ice-cold sodium bicarbonate solution, dried over magnesium sulfate and used directly for glucosylation. The yield is virtually quantitative. The amounts required for glycosylation are weighed out as 4-nitrobenzoate.

E) 4,5,7,8-Tetra-O-chloroacetate-2,3-dideoxy-2-fluoro-C-ft-D-manno-2-octulopyranosonic acid methyl ester (for example III) 25 - a) 4.5 7,8-Tetra-O-acetyl-3-deoxy-D-manno-2-octulopyranosonic acid methyl ester

1.19 g of 4,5,7,8-tetra-O-acetyl-2-bromo-2,3-dideoxy-D-manno-2-octulosonic acid methyl ester are dissolved in 10 ml of acetone and the solution is cooled to 0. After addition of 2.5 ml 1M aqueous sodium bicarbonate solution, stirring is continued for an additional 30 min at 0 ° (thin layer control: dichloromethane / ether - 5/1).

The acetone is then evaporated and the residue is shaken with dichloromethane. The organic phase is washed with water, dried and 35.

evaporated into a syrup. Chromatography on silica gel (hexane / ethyl acetate = 2/1) gives the title compound. = 50.7 ° 8702 24 8 - 30 - * (c = 1.08 in chloroform).

b) 4,5,7,8-Tetra-O-acetyl-3-deoxy - / - - D-manno-2-0- (tetrahydropyran-2-yl) -2-octulopyranosonic acid ethyl ester 5 At a solution of 0.917 g 4, 5,7,8-tetra-0-acetyl-3-deoxy-D-manno-2-octulopyranosonic acid methyl ester in dichloromethane, 0.99 ml of dihydropyran, a catalytic amount of O-toluenesulfonic acid monohydrate and molecular sieve 4A are added. The mixture is then stirred at room temperature for 2 hours.

The acid is then neutralized by adding sodium bicarbonate solution and the solid is filtered off. The filtrate is concentrated by evaporation and the residue is chromatographed on silica gel (hexane / ethyl acetate = 3/2). The title compound is obtained. / & (/ = 69.5 ° (c = 1.26 in chloroform).

C) 3-Desoxy-K-D-manno-2-Q- (tetrahydropyran-2-yl) -2-octulopyranosonic acid methyl ester

In a solution of 0.4 g of 4,5,7,8-tetra-O-acetyl-3-deoxy-b (-D-manno-2-0- (tetrahydropyran-2-yl) -2-20 octulopyranosonic acid methyl ester in 3 ml of methanol are added at 0 ° with a catalytic amount of metallic sodium under stirring.

After completion of the reaction (thin layer control: dichloromethane / methanol = 4/1), the mixture is neutralized with Amberlite IR 120 (H), the resin is filtered and washed with methanol, filtrate and wash solution are combined and evaporated. The title compound is obtained as a syrup. 75 ° (c = 1.05 in chloroform / methanol = 10/1). The compound is recognizable in a thin layer chromatography as a 1/1 mixture of the diastereomeric tetrahydropyranyl ether.

D) 4,5,7,8-Tetra-O-chloroacety1-3-deoxy-^ -D-manno-2-0- (tetr ahydrop3-ran-2-y 1) -2-octulopyranosonic acid methyl ester

6.5 ml of dry 2,6-utidine and 2.05 ml of triethylamine are added to a solution of 1.14 g of 3-deoxy-^ -D-manno-2-O- (tetrahydropyran-2-yl) - 2-octulopyranosonic acid methyl ester in 87 0 2 24 8 ♦ - 31 - 5 ml of dichloromethane. The solution is cooled to 0 ° and 3.5 g of chloroacetic anhydride are added. After stirring for an additional 20 hours at room temperature, the reaction mixture is cooled again, methanol is added to destroy excess reagent and the resulting mixture is evaporated to a syrup. The residue is dissolved in dichloromethane. The solution is washed successively with ice cold 2 M hydrochloric acid, water and dilute sodium bicarbonate solution, dried over magnesium sulfate and concentrated by evaporation.

The product is purified by chromatography on silica gel (hexane / ethyl acetate * 2/1) and the product is obtained as a syrup.

The two diastereomeric tetrahydropyranyl ethers can be separated chromatographically. The fast eluting component has t = 77.2 ° (c 0.5 in chloroform); the other component has / = * 7 ^ = 56.4 ° (c = 0.47 in chloroform) 15 e) 4,5,7,8-Tetra-Q-chloroacyl-1-3-deoxy-tt-D-manno- 2-octulopyranosonic acid methyl ester

A solution of 1.90 g of 4,5,7,8-tetra-0-chloroacetyl-3-deoxy-2-0- (tetrahydropyran-2-yl) - & C-D-manno-2-octulopyranosonic acid methyl ester in 50 is heated ml of glacial acetic acid up to 45 °. 5 ml of water are then added dropwise to the mixture and kept at 45 [deg.] For 5-6 hours (thin layer control: dichloromethane / methanol = 50/1). The mixture is then freeze-dried and the residue is diromatographed on silica gel (hexane / ethyl acetate = 2/1). The title compound is thus obtained. fo (7 ^ = 40.8 ° (c = 0.93 in chloroform).

F) 4,5,7,8-Tetra-0-chloroacetyl-2,3-didesoxy-2-fluoro -, (° -D-manno-2-octulopyranosonic acid methyl ester

323 mg of 4,5,7,8-tetra-O-chloroacetyl-3-deoxy-D (-D-manno-2-octulopyranosonic acid methyl ester) are dissolved in 50 ml of toluene and the solution is cooled to -60 °. A solution of 0.5 ml of diethylamino sulfur trifluoride in 2 ml of toluene is added and the mixture is stirred at -60 [deg.] for 10 minutes, then ice water is added and the mixture is treated with aqueous sodium bicarbonate solution and shaken with dichloromethane. washed with water, dried and concentrated by evaporation The syrupy residue is chromatographed on silica gel 870 2 2 * 8 - 32 - (hexane / ethyl acetate = 2/1) to give the title compound, which is pure on thin layer chromatography (dichloromethane / ether = 7/1), δ (^ p = 57.3 ° (c = 0.51 in chloroform).

5 F), 5-Didesoxy-7,8-Q-isopropylidene-4-O-methyl- (3-deoxy-4,5; 7,8-di-O-isopropylidene-06-D-manno-2- octulopyranosyl) acidic methyl ester-b 1 -D-manno-2-octulopyranosyl / acid acid methyl ester (for example IV) 10 a) (Ally1-3,5-dideoxy-4,7,8-tri-O-acetyl-UD-manno- 2-octulopyranoside) onacid methyl ester and (ally1-3,5-di-deoxy-4,7,8-tri-Q-acetyl-ft-D-manno-2-octulopyranoside) onacid methyl ester

A solution of 7.5 g (3,5 di-deoxy-2,4,7,8-tetra-O-acetyl - ((- D-manno-2-octulopyranoside) acid, methyl ester in 30 ml dichloromethane is left React at 0 ° with 10 g of TiBr 2 and keep overnight at 4 ° The solution is diluted with chloroform, extracted with ice-cold sodium bicarbonate solution, dried and evaporated, the yellowish syrup thus obtained is dissolved in nitromethane and added dropwise 0 ° a suspension of 5 ml of allyl alcohol, 4.5 g of Hg (II) cyanide and 3 g of molecular sieve 3A in nitromethane After 10 hours, the reaction mixture is filtered, concentrated by evaporation and chromatographed on silica gel using toluene / ethyl acetate. (3/1) Concentration of the fractions with R = 0.46 (toluene / ethy1-

Jt - .20 acetate = 2/1) gives the title compound as a syrup. T & I-q = 70.2 (c = 0.9 in chloroform).

b) (Ally1-7,8-0-carbony1-3,5-dideoxy-^ -D-30 manno-2-octulopyranoside) on acid methyl ester and (allyl-7,8-0-carbonyl-3,5-dideoxy-0 (- manno-2-octulopyranodide) onacid methyl ester.

A solution of 4.3 g (allyl-3,5-di-deoxy-4,7,8-tri-O-acetyl-OC-manno-2-octulopyranoside) onacid methyl ester and methyl (allyl-3,5- the deoxy-4,7,8-tri-O-acetyl-(-D-35 manno-2-octulopyranoside) onacid methyl ester in 50 ml of absolute methanol 870 2 ^ 48 - 33 - is added for 3 hours at room temperature with a solution of 70 mg of sodium stirred in 10 ml methanol, with ion-exchange resin • f

Dowex 50 (H) neutralized, filtered and concentrated by evaporation. The residue is taken up in absolute pyridine, treated with 5 g of p-nitrophenylchloroformic acid ester and stirred for 20 hours. The solution is concentrated on evaporation and the residue is taken up in ethyl acetate, extracted with sodium bicarbonate solution, dried concentrated by evaporation. Chromatography on silica gel using ethyl acetate 10 as an eluent gives the title compound as a syrup.

= + 23.2 ° (c = 1.4 in chloroform) and / d / D = + 53.7 (c = 1.0 in chloroform) c) ^ Allyl-7.8-0-carbonyl-3.5 -didesoxy- 4-O-methyl- (3-deoxy-4,5,7,8-tetra-O-acetyl-O [-D-manno-2-octulo-15-pyranosyl) -acid methyl ester & 6 -D-manno- 2-octulopyranoside / onacid methyl ester

A solution of 1.7 g of KDO bromide in absolute nitromethane is added dropwise to a suspension of 0.58 g (ally 1-7.8-0-carbony 1-3, S-dideoxy - ^ - D-maxmo- 2-octulopyranoside) onacid methyl ester, 1.25 g of Hg (CH) 2 and 1 g of molecular sieve 3A and stirring at room temperature for 15 hours. The reaction mixture is filtered, concentrated by evaporation and purified on silica gel using toluene / ethyl acetate (2/1). The title compound is obtained as a colorless syrup. = + 90.3 (c = 2.2 in chloroform).

8 7 0 2 2 4 8 - 34 - * d) / Allyl ~ 3,5-dideoxy-7,8-0-isopropylidene-4-0-methyl-3-deoxy- (4,5: 7,8-di -O-isopropylidene-^ -D-manno-2-octulopyranosyl) -acid methyl ester or -D-manno-2-octulopyranoside 7-acid methyl ester 5 Matt-treats a solution of 1.24 g /, allyl-7,8-0-carbony1- 3,5-dideoxy-4-0- (3-deoxy-4,5,7,8-tetra-0-acetyl-O-D-manno-2-octulopyranosyl) -oic acid methyl ester * K -D-manno- 2-octulopyranoside / acid methyl ester in 30 ml absolute methanol with a solution of 50 mg sodium in 50 ml methanol and stir at room temperature for 10 60 min. After neutralization with Dowex 50 (H +), the mixture is filtered and concentrated by evaporation.

The residue is dissolved in 10 ml of absolute dimethylformamide, treated with 10 mg of p-toluenesulfonic acid and 5 ml of acetone dimethyl acetal and stirred at 60 ° under slight vacuum for 48 hours. After evaporation of the solution, chromatography on silica gel is performed using toluene / ethyl acetate (1/1). The title compound is obtained as a colorless syrup. 54.9 ° (c = 1.2 in chloroform).

E) ^ 3> 5-Didesoxy-7,8-0-isopropylidene-4-0- (3-deoxy-4,5: 7,8-di-0-isopropylidene-Q {-D-manno-2- octulopyranosyl) onacid methyl ester - ^ - D-manno-2-octulopyranosyl / onacid methyl ester

A solution of 400 mg / allyl-3,5-dideoxy-7,8-0-isopropylidene-4-0- (3-deoxy-4,5: 7,8-di-25-isopropylidene-c () is treated D-manno-2-octulopyranosyl) acid methyl ester-o ((- D-manno-2-octulopyranoside 7oic acid methyl ester in 10 ml absolute tetrahydrofuran with 10 mg /ίr(COD)/PCH2 "(CgH,-)2?2.?2I > F6, which has been activated with hydrogen and stirred at room temperature for 90 min. The solution is diluted with dichloromethane, extracted with saturated sodium bicarbonate solution, dried and concentrated by evaporation. The residue is dissolved in 25 ml of tetrahydrofuran / water. (4/1) dissolved, treated with 250 mg of iodine and stirred for 20 min at room temperature After dilution with 100 ml of dichloromethane, extraction is effected with 5% sodium bicarbonate solution and the mixture is neutralized with sodium bicarbonate solution, dried and concentrated by evaporation Chromatography on silica gel with toluene / ethyl acetate (1 / 1) gives the title link.

870 2 24 8 -35- 1 H NMR: 4.52 (ddd, 1H, J4I 5, V8.0, J4I ^ * / 3.7, ^,, ^. 2.5.11-4 ^ 54.38 (m, tH, J4 5an / 11.0, J4 5e / v4.0, J4 ^ 5.0, ^ ^ 11.0, H-4); 4.38 (m, 1H, J71 gtA / 7.5 , Jyf gtaA / 6.0 »J ^, g, ^ / v4.5, H-7 *) j4.28 (dd, 1H, t ^ 1 ^ 2.0, J5t * 4.8.0, H- 5f); 4.14 (dd, 1H, Jg, a 8lb / V / 9.0, H-8 »a); 4.04 (dd, ΙΗ, Η-8'b); 5 3.99 (dd .1H, Jga gb / V7.5, Jga 7 ^ v6.0, H-8a); 3.91 (ddd, 1H, J? Gb / \ / 4.5, J? 6 / V7.5, H- 7); 3.80 (dd, 1H, H-8b); 3.80 (ddd, 1H, J6 ^ 2.0, ^ ^ λ / Μ, Ο, H-6); 3.76 (d, 1H , J3a QgA / 2.0); 3.82 (s, 3H, CH3OCO); 3.78 (s, 3H, CH3, OCO); 3.58 (dd, 1H, H-6T); 2.99 (dd, 1H, J,, A / 15.5, H-3e '); 2.06,

J J djJ C

(ddd, 1H, J3e 3a / V 12.5, J3e 5e / v 1.7, H-3e); 1.94 (dd, 1H, H-3a); 1.92 (ddd, 10 1H »J5e 5aA / 12.5, H-5e); 1.79 (dd, 1H, H-3'a); 1.44,1,38,1,36,1,32 (s, 18H, CHg); 1, 29 (t, 1H, H-5a).

G. 2,3-Di-O-benzyl-5-O- / 3-deoxy-4,5: 7,8-diisopropylidene-Q (.- D-manno-2-octulopyrauosyl) -acid methyl ester 7 ~ D ~ 13 arabinofuranose for Example V) a) 1,5-Di-0-acetyl-2,3-di-0-benzyl-D-arabinofuranose

A solution of 10 g of 2,3,5-20 tri-O-benzyl-1-O-p-nitrobenzoyl-D-arabinofuranose in J 50 mg of dry acetic anhydride is cooled to 10 °. A mixture of 15 ml of acetic anhydride and 100 ml of 95-97% sulfuric acid is added and the mixture is left at room temperature for 30 min. After cooling to 10 ° again, a few drops of sulfuric acid are added 23 and the mixture is left to stand at room temperature for 30 min. The reaction mixture is carefully poured into ice water and stirred for 10 min.

Maine neutralizes with saturated cooled sodium hydrogen carbonate solution. The water phase is extracted at pH 7.5-8 with 3 times 100 ml of dichloromethane. The organic phase is dried over magnesium sulfate and concentrated by evaporation. The title compound is obtained after chromatography on silica gel. = 0.39 (toluene / ethyl acetate 5/1).

b) 5-0-Acetyl-t-chloro-1-deoxy-2,3-di-Q-benzyl-Ci-D-arabinofuranose

1 g of 1,5-di-O-acetyl-2,3-di-O-benzyl-D-arabinofuranose is dissolved in 5 ml of glacial acetic acid and 20 ml of HCl gas saturated with glacial acetic acid are added at room temperature.

870 2 24 8 ι - 36 -

After 20 min, the mixture is treated with 35 ml of dichloromethane and 100 ml of ice water and shaking. The water phase is washed with 35 ml of dichloromethane, the combined organic phases are vigorously washed with 40 ml of cold saturated sodium hydrogen carbonate solution and dried over anhydrous magnesium sulfate. Concentration is then carried out under vacuum to obtain the title compound as a syrup.

Rj = 0.33 (toluene / ethyl acetate = 5/1).

c) Ally 1-5-0-acety 1-2,3-di-O-benzyl-10-arabinofuranoside

The following ingredients are placed in a round bottom flask under an argon atmosphere: 40 ml of dichloromethane (dried over P 2 O 3), 1.2 g of calcium sulfate (powdered, dried under vacuum at 120 °), 4.5 g of silver carbonate (under vacuum over calcium 15 sulfate dried) and 0.4 ml allyl alcohol. With the exclusion of light, 0.9 g of 5-O-acetyl-1-chloro-1-deoxy-2,3-di-O-benzyl-O-D-arabinofuranose dissolved in dried dichloromethane are added dropwise over 45 min. to. The reaction mixture is then stirred at room temperature for 18 hours. The resulting suspension is filtered over celite, washed once with dichloromethane and concentrated in vacuo. The title compound is obtained by chromatography on silica gel. = 0.54 (toluene / ethyl acetate - 5/1).

25 d) Ally1-2,3-di-O-benzyl- (λ-D-arabinofuranoside

2 g of allyl-5-0-acetyl-2,3-di-O-benzyl-D-arabinofuranoside are dissolved in 100 ml of methanol (freshly distilled over Mg curls). 2 ml of standard solution 30 (5 ml of methanol, 50 mg of metallic sodium) are then added and the mixture is allowed to stand at room temperature for 1 hour. Neutralizes with Dowex ^ 50-H, filters the resin and concentrates the solution by evaporation. The syrup is taken up in 10 ml of dichloromethane and crystallized with pentane. Mp. 61-62 °, Rf = 0.43 (toluene / 35 ethyl acetate = 2/1).

870 2 24 & t - 37 - e) Ally1-2,3-di-0-benzyl-5-0- (3-deoxy-4,5,7,8-tetra-0-acetyl-D-manno-2 -octulopyranosyl) onacid methyl ester-D-arabinofuranoside:

3.6 g of allyl-2,3-di-O-benzyl-O <-5 D-arabinofuranoside are dissolved in 40 ml of nitromethane (dried on molecular sieve 3A). 15 g of powdered molecular sieve (dried at 160P / 13 Torr) and 3.15 g of mercuricyanide (powdered, dried in vacuo on calcium filtrate) are added. 6.5 g of 3-deoxy-4,5,7,8-tetra-10-acety1-06-D-manno-2-octulopyranosyl bromide) acid methyl ester (dried in vacuo and in 12 liters) are added dropwise over a period of 6 hours. ml of absolute nitromethane dissolved) and allow the mixture to stand for an additional 18 hours. It is then diluted with 130 ml of dichloromethane, the resulting mixture is stirred for 30 hours, the solid is filtered off over celite and the solution is concentrated under vacuum to a syrup. It is separated by column chromatography (silica gel, gradient: toluene / ethyl acetate = 7 / 1- * 2/1).

The fractions with = 0.35 are collected and concentrated by evaporation. They contain the desired ketoside mixture as well as unreacted alcohol. Dissolve the well dried syrup in 40 ml of dry pyridine and drop 6 ml of acetic anhydride at ice bath temperature. After 6 hours, the mixture is treated at 0 ° with 5 ml of methanol. 30 min. Later, it is concentrated under vacuum, the remaining solvents are expelled with toluene and the residue is taken up in chloromethane. This solution is shaken with sodium bisulfate, sodium bicarbonate solution and water and dried over magnesium sulfate. The residue obtained after evaporation is chromatographed on silica gel (gradient: toluene / ethyl acetate = 4/1 2/1). The acetylated aglycone elutes at R = 0.66 (toluene / ethyl acetate = 2/1). The ketoside mixture elutes at 30 Rf = 0.35.

f) Ally1-2,3-di-0-benzy1-5-0-ZT3-deoxy-0 (, β-D-manno-2-octulopyranosyl) onacid methyl ester / -g ^ -D-arabino-furanoside: 35 Menlost7, 07 g ally1-2,3-di-0-benzy1-5-0- / 4,5,7,8-tetra-0-acetyl-3-deoxy - $, f ^ -D-manno-2-octulopyranosyl) -oic acid methyl ester / 4 ^ -D-arabinofuranoside at room temperature in g 7 λ * o 4 q - 38 - * absolute methanol. 2 ml of 0.1 N sodium methanolate are added thereto. After 1 hour, neutralization is effected with Dowex 50H +, the resin is filtered off and the solution is concentrated by evaporation. The title compound is obtained as a syrup. = 0.08 (toluene / ethyl acetate = 1/1).

g) Allyl-2,3-di-O-ben2yl-5-O-Zj (4.5: 7.8-di-O-carbonyl-3-deoxy-O-manno-2-octulopyranosyl) onacid methyl ester E7 ~ Q ^ -D-arabinofuranoside and ally1-2,3-di-0-benzy1-5-0 - / ^, 5: 7,8-di-0-carbonyl-3-deoxy- | 4-D-manno-2- octulopyranosyl) onacid methyl ester 7 “A Λ ^ -D-arabinofuranoside:

5.5 g of allyl-2,3-di-O-benzyl-5-O- / (3-deoxy-O /, ft-D-manno-2-octulopyranosyl) -acid methyl ester 7-d-D-arabinofuranoside are dissolved in 350 ml of dry pyridine and cools the solution to -35 °. Then it is added dropwise under vigorous stirring.

add a solution of 1.2 ml of chloroformic acid trichloromethyl ester in 40 ml of dry benzene. After 1.5 hours at -35 ° C, the mixture is allowed to warm to room temperature and the completion of the reaction is determined by thin layer chromatography (toluene / ethyl acetate = 1/1). R ^ values: starting material 0.08; OtKDO 0 (ara (di-O-carbonyl) = ^ 0.31). |) Κ00Γ ^ ara (di-O-carbonyl) = 0.15.

After completion of the reaction, 5 ml of ethanol are added dropwise. The mixture is concentrated under vacuum and concentrated again three times with toluene.

A dichloromethane solution of the residue is washed with sodium hydrogen carbonate solution and water and dried over magnesium sulfate and the solvent is evaporated. Separation of the KD0 anomers over silica gel (toluene / ethyl acetate = 2/1) affords the title compound.

^ 20 OC KDO Disaccharide: Silj. = -7.1 ° (c = 4.8 in chloroform).

30 U20 yy KDO Disaccharide: = 5.2 ° (c = 2.1 in chloroform): mp 146-148 °.

h) Allyl-2,3-di-O-benzyl-5-O-2'3-deoxy-O (-D-manno-2-octulopyranosyl) -acid methyl ester / -Qd-β-arabinofuranoside: 2 g are dissolved ally1-2,3-di-0-benzyl-5 ~ 0- / T4,5: 7,8-di-0-carbonyl-3-deoxy- (X-D-manno-2-octulopyranosy1) on-acid methyl ester 7- ^ -D-arabinofuranoside at room temperature in 87 DEG-24 DEG-39-40 ml of dry methanol. 2 ml of 0.1 N sodium methanolate are added.

Mai neutralizes after 3 hours with Dowex 50H +, filters and removes the solvents under vacuum. R ^ = 0.08 (toluene / ethyl acetate = 1/1).

5 i) Allyl-2,3-di-O-benzyl-5-O- (3-deoxy-4,5-yl-diisopropylidene-1 ^ -D-manno-2-octulopyranosyl) -acid methyl ester / ^ - D-arabinof uranoside;

0.8 g of allyl-2,3-di-O-benzyl-5-O- / f (3-deoxy-0 (-D-manno-2-oc tu 1 opyr ahos y 1) is dissolved in our / - C (-D-10 arabinofuranoside in 100 ml dimethylformamide (distilled over molecular sieve 4A) and add a catalytic amount of toluene-4-sulfonic acid. 0.49 ml 2-methoxypropylene is injected directly into this solution and the reaction vessel is closed immediately.

After 5 hours, 0.49 ml of 2-methoxypropylene is added and the mixture is left to stand for 18 hours. Neutralize with solid sodium carbonate, remove the solvent in vacuo, and obtain the title compound by column chromatography over silica gel (toluene / ethyl acetate = 2/1).

20

Rf ^ ° 43; = -9.3 (3.3 in chloroform).

20 j / k) 2,3-Di-O-benzyl-5-0- / T (3-deoxy-4,5; 7,8-diisopropylidene-k / -D-manno-2-octulopyranosyl) onacid methyl lesterJ-D-arabinofuranose:

1.6 g of allyl-2,3-di-O-benzyl-5-O-1 (3-deoxy-4,5: 7,8-diisopropylidene - - - D-manno-2-octulopyranosyl) -25 are dissolved unacid methyl ester / -D-arabinofuranoside in 40 ml of dry tetrahydrofuran. 3.2 g of 1,5-cyclooctadiene-bis- / methyl diphenylphosphine iridium hexafluorophosphate are added. The device is carefully rinsed with argon (99.996%) and the solution is stirred under oxygen-free argon for 3 min. The catalyst is then activated by purging with hydrogen 2-3 times. 5 Min. later it is again sprayed with argon and the mixture is stirred at room temperature for 2 hours. The reaction mixture is added to 150 ml of tetrahydrofuran / water (4/1), 1.5 g of iodine is added and the whole is left to stand at room temperature for 5 min. It is diluted with 1.2 l of water, shaken with chloroform, the organic phase is washed twice with 500 ml of 5% sodium pyrosulfite solution each time, dried over magnesium sulfate and concentrated in vacuo. The resulting syrup is separated by column chromatography over silica gel (gradient: toluene / ethyl acetate = 870 2 24 8 - 40-3 / 1-V 1/1). lip = 0.58 (toluene / ethyl acetate = 1/2); = + 33.2 ° (c = 1.6 in chloroform).

H) Allyl-2,3-di-O-benzyl-5-O- / t3-deoxy - ((-5 D-manno-2-octulopyranosyl) -acid methyl estery-O (.- D-arabinofuranoside a) 2- Anisylideneamino-2-deoxy-D-glucose:

10 g of glucosamine hydrochloride are dissolved in 47 ml of 1N soda. 5.7 ml of anisaldehyde 10 are added dropwise to this solution, on which a crystalline precipitate appears. Mai keeps the reaction mixture at ice bath temperature for 1 hour. The crystals are filtered and washed with ice cold water and alcohol / ether (1/1) and then air dried. M.p .: 161-164 ° (dec.).

B) 2-Anisylideneamino-2-deoxy-1,3,4,6-tetra-O-acetyl-D-glucopyranose;

10 g of 2-anisylideneamino-2-deoxy-D-glucose are dissolved in 10 ml of absolute pyridine. Then there is added 60 ml mixture of pyridine and acetic anhydride (1/1) at ice bath temperature. 2 hours later, the mixture is allowed to warm to room temperature and left overnight. The mixture is then poured into ice water, on which the desired product immediately crystallizes out. The crystals are filtered and recrystallized from dichloromethane / pentane. M.p .: 180-182 °.

c) 2-Amino-2-deoxy-1,3,4,6-tetra-O-acetyl-D-glucopyranose. hydrochloride:

10 g of 2-anisylideneamino-2-deoxy-1,3,4,4,6-tetra-O-acetyl-D-glucopyranose are dissolved in 400 ml of acetone and heated to a reflux condenser. Then 1 mole equivalent of 5N hydrochloric acid is added dropwise, on which the title compound precipitates immediately. After cooling, 300 ml of diethyl ether are added, the mixture is stirred vigorously for 30 min and filtered. The crystals are washed with diethyl ether and dried in a desiccator on calcium sulfate.

M.p .: 220 ° (dec.).

8 7 0 2 3 A 8 - 41 - d) 2-Afflino-2-deoxy-1,3,4,6-tetra-Q-acetyl-D-glucopyranose:

Them dissolves 17.7 g of 2-amino-2-deoxy-1,3,4,6-tetra-O-acetyl-D-glucopyranose hydrochloride in a little water.

16.34 g of sodium acetate are added thereto and the mixture is stirred at room temperature for 30 minutes. The mixture is shaken 5 times, each time with 50 ml of dichloromethane, the organic phase is dried over magnesium sulfate and the solvent is evaporated in vacuo. The title compound is recrystallized from hot ethanol. Mp. 132-134 °.

10 e) N- (R) -Hydroxytetradecanoyloxy / suceinimide:

22 g of 3- (R) -hydroxytetradecanoic acid are dissolved in 1 l of ethyl acetate and cooled to 4 °. 10.35 g of N-hydroxysuccinimide and 18.54 g of dicyclohexylcarbodiimide are added with vigorous stirring, upon which dicyclohexylurea begins to precipitate. After 3-4 hours, the mixture is warmed to room temperature and stirred overnight. The cyclohexylurea is then filtered off and the solvent is evaporated. The syrupy residue is dissolved in a little methanol at 40 ° and slowly cooled to 4 °, whereupon the title compound crystallizes out. Mp. 103-104 °.

f) 2-Desoxy-2- / 3- (R) -hydroxytetradecanamido./1-1,3,4,6-tetra-O-acetyl-D-glucopyranose;

10 g of 2-amino-2-deoxy-1,3,4,6- are dissolved. 25 tetra-O-acetyl-D-glucopyranose in 60 ml freshly distilled tetrahydrofuran. 14.7 g of N- (3) (R) -hydroxytetradecanoyl-oxy / succinimide are added thereto and the mixture is stirred at reflux for 48 hours. The reaction mixture is then concentrated in vacuo. The title compound is obtained from the residue by chromatography on silica gel.

R 3 = 0.47 (toluene / ethyl acetate-1/2), m.p. 143-145 °.

g) Benzyl-2-deoxy-2- ^ 3- (R) -hydroxy-tetradecanamido -3.4.6-tri-O-acetyl-tS -D-glucopyranoside:

A suspension of 2.03 g of iron (III) -35 chloride (anhydrous) and 3.9 g of anhydrous calcium sulfate in 65 ml of dry dichloromethane is stirred for 10 min. Then 7.8 ml of freshly distilled benzyl alcohol and 4.68 g of 2 are added. deoxy-2 - (R) -hydroxytetradecanamid (8702248) -42, 1,3,4,6-tetra-O-acetyl-D-glucopyranose and stir the suspension at room temperature for 96 hours. The mixture is then poured into cold saturated sodium bicarbonate solution and extracted 6 times with 100 ml of dichloromethane each time. The organic phase is washed with water, dried over magnesium filtrate and concentrated in vacuo. The title compound is obtained by chromatography on silica gel (gradient) :( toluene / ethyl acetate = 2/1 - 1/2 900 ml). Mp. 150-152 °; Rf = 0.48 (toluene / ethyl acetate = 1/2).

H) Benzyl-2-deoxy-2- / 3- (R) -hydroxytetra-decanamido / -fi-D-glucopyranoside:

4 g of Benzyl-2-deoxy-2- / 3- (R) -hydroxytetradecanamido-3,4,6,6-tri-O-acetyl-'-'-D-glucopyranoside 15 are dissolved in 100 ml of dry methanol. Then 3 ml of a solution of 5 mg of metallic sodium in 5 ml of methanol is added. One hour later, the mixture is neutralized with Dowex 50H +, the resin is filtered off and the solution is concentrated by evaporation. The title compound has R = 0.71 (chloroform / methanol / glacial acetic acid / water = 16/5/1/1).

20 1 fc 7 0 2 2 4 8

Benzyl-2-deoxy-4-0-diphenylphosphono- 3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamidq7-f--D-glucopyranoside (for example V): 25 a) Benzyl-2-deoxy- 2- / 3- (R) -hydroxytetra-decanamido7-4,6,0-isopropylidene-ft-D-glucopyranoside:

3 g of benzyl-2-deoxy-2 - / - (R) -hydroxytetradecanamidoj-D-glucopyranoside are dissolved in 40 ml of fresh 2q distilled dimethylformamide. 1 to 2 spatula dots of toluene-4-sulfonic acid monohydrate and 1.3 ml of isopropenyl methyl ether are then added. After standing at room temperature for 2 hours, neutralization is effected with solid sodium hydrogen carbonate (stirring for 10 min). The solvent is evaporated under high vacuum and the residue is concentrated again with 3 x 20 ml of toluene. The residue is dissolved in dichloromethane, the undissolved solid is filtered off and the solution is washed with water, dried over magnesium sulfate and concentrated by evaporation.

The residue is dried under vacuum to an amorphous powder. R ^ = 0.43 (ethyl acetate).

- 43 - b) Benzyl-2-deoxy-4,6-Q-isopropylidene-3-O-tetradecanoyl-2- £ 3- (R) -tetradecanoyloxytetradecanamido7-p-D-glucopyranoside:

3.4 g of Benzyl-2-deoxy-2-Z3-5 (R) -hydroxytetradecanamido7-4,6-0-isopropylidene) ¾-D-glucopyranoside are dissolved in 50 ml of a mixture of a mixture of dry dichloromethane and freshly distilled pyridine (2/1). A spatula tip of dimethylaminopyridine is added and the mixture is cooled to ice bath temperature. A mixture of 5.2 ml of tetradecanoyl chloride and 5 ml of dichloromethane is then added dropwise and stirred at ice bath temperature for 48 hours. Then 5 ml of methanol is added dropwise, followed by dilution with 25 ml of toluene and concentration in vacuum. Vacuum concentration with toluene is carried out several times to remove the residual pyridine and chromatographed on silica gel (toluene / ethyl acetate = 10/1). ; Rf = 0.32.

c) Benzyl-2-deoxy-3-0-tetradecanoyl-2-Z3- (R) -tetradecanoyloxytetradecanamid / fr-D-glucopyranoside 20 0.5 g of benzyl-2-deoxy-4,6-0-isopropylidene -3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetra-decanamido / -pD-glucopyranoside for about 7 min in 10 ml ice cold 90% trifluoroacetic acid. The reagent and solvent are then removed in vacuo, the residue is dissolved in dichloromethane and the solution is shaken with saturated sodium bicarbonate solution. The organic phase is dried over magnesium sulfate and the product is obtained after chromatography on silica gel (toluene / ethyl acetate = 2/1-ethyl acetate) ). R ^ = 0.22 (toluene / ethyl acetate = 1/1).

D) Benzyl-2-deoxy-3-Q-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamido7-6-Q-triphenylmethyl-R-D-glucopyranoside:

15 g of benzyl-2-deoxy-3-O-tetradecanoyl-2-Z3- (R) -tetradecanoyloxytetradecanamido / - ^ -D-35 glucopyranoside are dissolved in 1.3 l of acetic nitrile and the solution is heated to 60 °. Thereafter, 4 times 5 g of tritylpyridinium tetrafluoroborate is added in portions. The product formation is followed by thin layer chromatography (R.70 = 0.43 in toluene / ethyl acetate = 5/1).

Immediately after completion of the reaction, the solvent is evaporated, the residue is dissolved in dichloromethane and the solution is washed with water, dried over magnesium sulfate and purified by column chromatography (toluene / ethyl acetate = 11/1).

e) Benzyl-2-deoxy-4-Q-diphenylphosphono-3-tetradecanoyl-3-ir3- (fc) -tetradecanoyloxytetradecanamido. 7-6-0-triphenylmethyl-j-D-glucopyranoside: 10 330 mg Benzyl-2 -deoxy-3-O-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanamid-6-6-triphenylmethyl-1-D-glucopyranoside is carefully dried and dissolved in 50 ml of dry dichloromethane. 107 mg of dimethylamino-pyridine, 1 ml of absolute pyridine and 300 ml of diphenylphosphoric acid chloride are then added and the mixture is stirred at room temperature for 90 hours.

The mixture is then diluted with dichloromethane, shaken with saturated sodium bicarbonate solution, the organic phase is dried over magnesium sulfate and the product is obtained by chromatography on silica gel (gradient: toluene / ethyl acetate = 1/1 * 9/1).

20 = 0.34 (toluene / ethyl acetate = 10/1).

f) Benzyl-2-deoxy-4-0-diphenylphosphono-3-0-tetradecanoyl-2- / 3- (R) -tetra-decanoyloxytetradecanamido7-ft-D-glucopyranoside: 120 mg of benzyl-2-deoxy- are dissolved 4-0-diphenylphosphono-3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecana-midi / -6-O-tr-enylmethyl-1-D-glucopyranoside in dichloromethane. At ice bath temperature, 90% trifluoroacetic acid is added until the yellow color of the solution no longer disappears. Then the solution is poured into saturated sodium hydrogen carbonate solution and shaken. The organic phase is dried over magnesium sulfate, the residue is concentrated and chromatographed on silica gel (Gradient: toluene / ethyl acetate = 6/1-k 3/1, 500 ml each). R ^ = 0.45 (toluene / ethyl acetate = 2/1).

35 870 2 24 8 * - 45 - K) (3-Desoxy-4,5,7,8-tetra-O-chloroacetyl-D-manno-2-octulopyranosyl bromide) -oic acid benzyl ester for Example VI: a) 3-Desoxy- 2-0- (tetrahydropyran-2-yl) -D-5 manno-2-octulopyranosonic acid benzyl ester:

26.9 ml of 0.2M potassium hydroxide are added to a solution of 1.14 g of 3-deoxy-2-0- (tetrahydropyran-2-yl) -D-manno-2-octulopyranosonic acid methyl ester in 20 ml of dry dioxane and stirring mixture for 1 hour at room temperature. The reaction is followed by thin layer chromatography (dichloromethane / methanol = 2/1). The mixture is then treated with Amerblite IR-120 (H) to remove the base. The resin is filtered off and washed with methanol / water (10/1). Filtrate and wash solutions are combined and concentrated by evaporation. The residue is dissolved in 10 ml of Ν, Ν-dimethylformamide and left for 12 hours at room temperature with 0.373 g of potassium carbonate and 1.4 g of benzyl bromide. Ice water is then added and the product extracted with ethyl acetate and dichloromethane. The extracts are combined, dried over anhydrous sodium sulfate, concentrated by evaporation and the residue is chromatographed on silica gel (dichloromethane / methanol = 30/1). The product is a diastereomeric mixture of the tetrahydropyranyl ethers; the components migrate closely together on thin layer chromatography (dichloromethane / methanol = 3/1). The fastest migrating isomer has a melting point of 30 ° and a Ch (~ Jq - + 40 ° (c = 0.64 in dichloromethane) and the slowest migrating isomer has a melting point of 39-40 ° and a ^- ^ ~ + 44 ° (c = 2.2 in dichloromethane).

b) 2-O-Acetyl-3-deoxy-4,5,7,8-tetra-0-30-chloroacetyl-D-manno-2-octulopyranosonic acid benzyl ester:

Menlost dissolves 0.3 g of 3-deoxy-2-0- (tetrahydropyran-2-yl) -D-manno-2-octulopyranosonic acid benzyl ester, 1.9 ml of 2.6 lutidine and 0.59 ml of triethylamine in 5 ml of dry dichloromethane and slowly add 1 g of chloroacetic anhydride.

The mixture is stirred at room temperature for 18 hours. Then a small amount of ice is added and the solvent is removed in 870 2? ·: 8 - 46 - vacuum. Mai takes up the residue in dichloromethane and successively washes with 1M hydrochloric acid, water, aqueous sodium bicarbonate and water. After concentration by evaporation, the residue is chromatographed on silica gel (dichloromethane). The resulting intermediate is dissolved in 20 ml of acetic nitrile, 0.1 ml of 42% aqueous tetrafluoroboric acid is added and the mixture is stirred vigorously at room temperature for 15 minutes. Then 0.1 ml of triethylamine is added and the solvent is evaporated. The residue is chromatographed on silica gel (dichloromethane) to give the 2 O 2 H compound; / o (7 ^ = + 30 ° (c = 0.72 in dichloromethane).

This is dissolved in 4.5 ml of dichloromethane / 2,6-lutidine (2/1) and a solution of 0.1 ml of acetyl chloride in 3 ml of dichloromethane is added. The mixture is stirred overnight (20 °) and then poured into ice-cold sodium bicarbonate aqueous solution.

15

The product is extracted with dichloromethane, the extract is washed with 1M hydrochloric acid and water, dried and concentrated by evaporation.

The residue is chromatographed on silica gel (dichloromethane) and the amorphous title compound is obtained. = + 47 °.

C) (3-Desoxy-4,5,7,8-tetra-0-chloroacetyl-D-manno-2-octulopyranosyl bromide) acid benzyl ester:

A solution of 0.35 g of titanium tetrabromide in a mixture of 2.5 ml of dry dichloromethane and dry ethyl acetate (10/1) is added to a solution of 0.1 g of 2-0-acetyl-3-deoxy-4, 5,7,8-tetra-0-chloroethyl-D-manno-2-octulopyranosonic acid benzyl ester in 2 ml dry dichloromethane at 0 °. After the completion of the reaction (thin layer control: dichloromethane / ether = 15/1), 15 ml of dry dichloromethane are added and the mixture is treated with anhydrous sodium acetate. The suspension is filtered through celite and the filtrate is concentrated under vacuum at 20 °. The residue is freeze-dried from dry dioxane and used directly in the glycosidation reaction.

35 L) 3,5-Didesoxy-2,4,7,8-tetra-Q-acetyl-D-arabino-2-octulopyranosonic acid methyl ester: 87 0 2 24 δ 'ft - 47 -

A solution of 8 g of 2-deoxy-D-ribose and 50 mg of sodium bicarbonate in 43 ml of distilled water with soda is adjusted to pH = 11. 3 g of oxalacetic acid are added in portions and the pH is kept as close to 11 as possible for 3 hours, then 5 is heated. +

the solution to 50 ° and add Dowex 50-H resin to a pH

of about 5.7 has been reached. The mixture is kept at 50 ° for 2 hours, after which the pH rises to 6.2. The resin is then filtered off and the filtrate is placed on a column (80 x 3 cm) of Dowex-1x8 (bicarbonate form).

The column is washed with 500 ml of water and then eluted with a linear gradient (800 ml of water and 800 ml of 0.3 M ammonium bicarbonate).

Elution is then continued with a further 800 ml of 0.3 M ammonium bicarbonate.

Fractions of about 5 ml are collected throughout the elution.

Analysis by thin layer chromatography shows that the desired 3,5-dideoxy-D-arabino-2-octulosonic acid is present in fractions 175-220 (methanol / chloroform / water = 10/10/3). These fractions are combined, freed from bicarbonate using Biorex-70 (H +) resin, adjusted to pH 8.5 with a few drops of 30% ammonia and lyophilized. The lyophilized product is acetylated as described for 3-deoxy-D-manno-2-octulosonate (FMUhger, D.Stix and G. Schulz, Carbohydr. Res. 80, 191f \ 980j) and the resulting 3.5 -dideoxy-2,4,7,8-tetra-O-acety1-D-arabino-2-octulosonic acid also as described therein in the methyl ester. = 0.28 (toluene / ethyl acetate = 2/1).

25 870 2 24 8

Claims (15)

A chemical compound, characterized in that it has the formula 1, wherein represents a group of any one of formulas 2a-2k, 5 wherein R 1 represents hydrogen, a metal equivalent or alkyl, X and Y are the same or different and for oxygen or an imino group and R 2 and R 2 are the same or different Ί0 and represent an optionally substituted acyl group, wherein the compound may be in free form or in salt form. A compound according to claim 1, characterized in that R 1 represents a group of one of formulas 2a-2h as defined above 15. A compound according to claim 1, characterized in that a group of formula 2a, 2b, 2c, 2d or 2g, wherein R 1 represents hydrogen or alkyl, X represents oxygen, Y represents imino and R 2 and R 1 represents tetradecanoyl 2.0 optionally substituted with hydroxyl or tetradecanoyloxy. A compound according to claim 1, characterized in that XR 2 represents tetradecanoyloxy, Y represents R 3 -tetradeanoyloxytetradecanamido and R 1 represents a group of formula 2c, 2b or 2a, wherein R 1 represents methyl. 4 η - 48 - CONCLUSIONS " A compound according to claim 1, characterized in that (I) XR 2 and YR 1 are as defined in claim 4 and R 1 represents a group of the formula 2d or 2g, wherein R 1 represents methyl or a group of the formula 2a, wherein R 1 represents hydrogen, or (ii) XR 2 represents 3-tetradecanoyloxytetradecanoyl-oxy, R 1 represents a group of formula 2a, wherein R 1 represents hydrogen and YR 1 represents 3-hydroxytetradecanamido or 3-tetra-decanoyloxytetradecanamido. 870 2 24 8 - 49 - 6. A compound according to claim 4, characterized in that the 3-position of XR ^ or YR ^, when substituted, is in the 11 (R) "configuration and the glucopyranose ring of formula 1 is in the D-form. A compound according to claim 5, characterized in that the 3-position of XR ^ or YR ^, when substituted, is in the "(R)" configuration and the glucopyranose ring of formula 1 is in the D-form. 8. Chemical compound, characterized in that it has 6-0- (0 (-D-arabinofuranosyl) -2-deoxy-4-0-phosphono-3-0-tetradecanoy1-2- / 3- (R) - tetradecanoyloxytetradecanamidq / -D-glucopyranose, 2-deoxy-6-0-0 (-D-glucopyranosyl-4-0-phosphono-3-0-tetradeeanoyl- 2- / 3- (R) -tetradecanoyloxytetradecanamidq7-D-glucopyranose or 2-deoxy-6-0-methyl (3-deoxy-1b-D-manno-2-octulopyranosyl) -ono-4-0-phosphono-3-0-tetradecanoyl-2- / 3- (R) -tetradecanoyloxytetradecanami -do7-, 3-D-glucopyranose. 9. Chemical compound, characterized in that it is 2-deoxy-6- / 4-0- (3-deoxy-AD-manno-2-octulopyranosyl) -acid-methyl ester-3,5-dideoxy - ^ - D- arabino-2-octulopyranosylic acid methyl este7-4-0-phosphono-3-O-tetradecanoy1-2- / 3- (R) -tetradecanoyloxy-tetradecanamido / -D-glucopyranose, 25. Pj. 2-deoxy-6-0- / 5-0- (3-deoxy-t4-D-manno-2-octulopyranosyl) -acid methyl ester ^ 7 - <\ - D-arabinofuranosyl / -4-0-phosphono-3- 0-tetradecanoyl- 2- / 3- (R) -tetradecanoyloxytetradecanamido / -D-glucopyranose, 2-deoxy-6-0 -_ / (3-deoxy-tK.-D-manno-2-octulopyranosyl) -acid 7-2q 2 - / 3- (R) -hydroxytetradecanamida / -4-0-phosphono-3- / 3- (R) -tetradecanoyl-. oxytetradecanoyl / -D-glucose, 2-deoxy-6-0- / jt3-deoxy-t -D-manno-2-octulopyranosyl) unsaturated / - 4-0-fo s phono-3-0-tetradecanoy1-2- / 3- (R) -tetradecanoyloxytetradecanamida / - D-glucose, or 2-deoxy-6-0_Zj3-deoxy-Qi-D-manno-2-octulopyranosyl) impure ^ / - 4-0-phosphono-2- / 3 " (R) -tetradecanoyloxytetradecanamido / -3-0- / 3- (R) -tetra- 870 2 24 8 * * x -50-decanoyloxytetradecanoyl / -D-glucose. Compound according to claim 8 or 9, characterized in that it is in the form of an acid addition salt. Pharmaceutical composition, characterized in that it contains a compound of the formula 1 according to claim 1 in free form or in the form of a pharmaceutically acceptable acid addition salt, optionally together with a pharmaceutically acceptable diluent or carrier. 12. A method of using a chemical compound, characterized in that a compound of the formula 1 according to claim 1 is used as a medicine in free form or in the form of a pharmaceutically acceptable acid addition salt. 13. A method for using a chemical compound, characterized in that a compound of the formula 1 according to claim 1 in free form or in the form of a pharmaceutically acceptable acid addition salt is used as a modulator of nonspecific resistance to microorganisms. systemic treatment of immune response, in the enhancement of nonspecific immunity, as a means of treatment or supportive treatment of viral diseases, morbus Hodgkin and other malignancies, or as a means of preventing endotoxin shock. 14. Process for preparing a compound according to claim 1, characterized in that a) a compound of the formula 3, wherein R 2, X and Y have the above meaning and is a protecting group, is reacted with a compound of formula 4, wherein R 2 has the same meaning as R 2, but optionally hydroxyl groups which are present are in protected form and a group R 1 in R 1 represents a group R 2 which is alkyl or a protecting group and Z represents a cleaving group to give a compound 35 of the formula V in which R1, R25, R25, R, X and Y have the above meanings and a phosphate group in the compound 870 2 24 8 β-51 - having the formula 5, or b) a compound of the formula 6, wherein R 1, R 1, R 5> X and Y have the above meanings and Rg is a protecting group, react 5 with a compound of the formula h to give a compound of the formula 7, wherein R ^, R ^, R g, Rg, R *, X and Y have the above meanings, deprotected compounds obtained in protected form and recover the compound of the formula 1 in free form or in acid addition salt form. 15. New method, new chemical compound or new pharmaceutical preparation as described in the description and / or the examples. 15 -o-o-o-o- 8702248 £ CHjOH ° l'H? CHOH CHPH HONt _ $ ^ o \ S!> \ Ηο-λΧ> \ Hq / * R * x \ ^^ VnHH ° ^^ Vo- Η0 · ^^ - 0- Y R3 COORl) 0H 1 2a 2b HOI-feCp HQ CHjOH V ^ 0 'LSÏU »0 Γ * OH 2c Ηθ \ ^ - ^ 0 ro ^ \ s3S \ 2d & *> HOV ^^ ~ o- _ OH OH 2nd ch2oh ho' ^^^ ° \ HO | Hi0H Η0 \ ^ Ά fe-Λ ΗΟ · Α ^^ \ ~ · οοοΡή ΗΟ \ ^ Λ ^ / θ- $ oh 2f Ov_ CH, OH ^, CH2 HO I Ck I CHOH 2g \ Λ ^ ° \ ho \ ^^^ WCOORj, ^ .0 CH, 'HO II CHOH „\ 2 h Ηθ \ ^ --- \ ^ 0- 870 2 24 8 00R4 SANDOZ AG Basel, Switzerland CHêOH HO | r; CHeOH COOR ^ V - ^^ O- ^ CHOH CHjOH COORl, v \> ° \ HO I COORi, ---- A ~ -0- 2J CHOH COORi, CHOH 0 CH20H \ CHOH η H0 o ^ p ^ · \ COOR if Nsn ^ n \ / v®— □ WH, 2k ΗΟ ^ Λ ^ '° \ l'1 Rax \ ^^ c \ ^ ORr. ^ \ H ^ \ Yr Rt -z ΗΟ ^ Λ \ '* - ^^ \ 3. i ReX - ^^ - ^^ ORj 5 „n 9 CHOH YRi Rfc0 RjX' ^ w - ^ \\ ^ - ors Rt- Z yr3 6 ia l 'R' \? I, "B" ° "•« A -'- ^ wor, 87 0 2 24 8 IR "SANDOZ A.G. Basel, Switzerland