JPS60123492A - Phosphorylated glycosyl-amide, -urea, -carbamate and thiocarbamate - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides novel monophosphorylated 7'CN-glycosylamides, N-glycosylureas, N-glycodylcarbamates and N-glycofluthiocarbamates, their production methods, and medicaments. Regarding its use as.

The new compound has the general formula, where tR is an optionally substituted straight-chain or branched saturated, monounsaturated or polyunsaturated alkyl group, an optionally substituted saturated or monounsaturated alkyl group, or an optionally substituted saturated or monounsaturated alkyl group. Table 4 shows saturated or polyunsaturated cycloalkyl groups, or optionally substituted aryl or aralkyl groups. C) Xil: CH2,0,5SI (represents N-alkyl across H, the alkyl group may contain up to 20 carbon atoms, preferably 1 to 10 carbon atoms, R2 has the meaning of R1, and When X represents CH, it can additionally represent hydrogen gold.1 The radicals R3 are, independently of each other, hydrogen 1 an acyl group of 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms, ~20 alkyl groups, preferably 1 to 10 alkyl groups, 1 representing a silyl group and/or a phosphoric acid or phosphoric acid ester group, 1 and R4 representing hydrogen, methyl or 10H, -0-R'; , one of the groups Ha always represents a phosphoric acid or phosphate ester group.

The alkyl group R1 has up to 40 carbon atoms, preferably 20
up to 1 and very particularly preferably from 10 to 2 carbon atoms
It has 0 pieces.

Examples of saturated alkyl groups R1 are methyl 1 ethyl n-globil, inglovir, n-butyl, inbutyl 1 n-pentyl, n-hexyl, n-hebutyl, n-octyl, n
-nonyl, n-decyl, n-undecyl, n-dodecyl, n-tridecyl (n-f) labucyl, n-/J!
ntadecyl 1n-hexadecyl, n-heptadecyl, n
-octadecyl, n-nonadecyl, eicosyl, tocosyl, tetracosyl, triacontyl 1 ethylpentyl,
Methyldecyl, inflovirdecyl, methyltridecyl t-pentylhexadecyl, 1-dodecylhexadecyl,
2'-dodecylhexadecyl, 5-dodecylhexadecyl, 1-hexadecyloctadecyl, 2-hexadecyloctadecyl, 3-hexadecyloctadecyl 1.4-
Hexadecyl octadecyl 11-octadecyl eicosyl and 2-octadecyl eicosyl.

Examples of unsaturated alkyl groups R1 are ethynyl, prop-1-enyl, furog-2-enyl, isofthynyl, boudo-1-enyl.
Erhept-2-enyl 1-pent-1-enyl, bent-2-enyl, bent-3-enyl, bent-4-enyl, hex-1-enyl, hex-2-enyl, hex-5-enyl, hex-4-enyl, hex-5-enyl, -rp-1-enyl, dec-5-enyl, thic-
9-enyl, hegetadecou-8-enyl, but-1,5-
Genyl, penta-1,5-genyl, penta-1,4
-genyl, heptadecane-Q, 11-genyl and heptadecane-8,11,14-)genyl.

Generally, among unsaturated groups, long-chain groups, especially from 10 carbon atoms to
Preference is given to 20 monounsaturated or diunsaturated alkenyls.

The unsaturated hydrocarbon radicals can be in the form of pure cis- or trans-isomers or as a mixture of isomers.

The cycloalkyl group R1 is preferably a group of 5 to 7 carbon atoms. Examples of cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl, cycloalkyl, cycloheptyl, cyclohexylcyclohexyl and cyclopentylcycloheptyl.

Examples of unsaturated cyclic radicals R1 are cyclopentenyl, cyclohexenyl and cycloheptenyl. Preferred substituents for cycloalkyl groups are alkyl groups of up to 8 carbon atoms.

Examples of nuclei that may be mentioned are: methyldicyclopentyl 1-ethylcyclopentyl, n-7'-bicyclopentyl, isoprogylcyclopentyl 1-butylcyclopentyl, octylcyclohentyl, methylcyclohexyl, ethylcyclohexyl, propylcyclohexyl isomer. Butylcyclohexyl, hexylcyclohexyl, decylcyclohexyl) cyclopentylmethyl, cyclopentyl ether 1 cyclopentylpropyl, cyclopentyl butyl, cyclopentylpentyl 1 cyclopentylhexyl) cyclopentyl octyl, cyclopentyl decyl - cyclohexylmethyl 1 cyclohexyl ethyl 1 cyclohexylpropyl) cyclohexyl butyl 1 cyclohexyl, cyclohexyl xyldecyl 1 cyclopentylcyclohexylethyl, cyclohexylcyclopentylethyl and hiscyclohexylcyclohexylethyl.

The substituents can be arranged in each case in the cis- or trans-position.

The aralkyl group R1 can take many forms. Thus, t ore 1 preferably has 6 or 10 carbon atoms.
An aryl group consisting of 1 or more may be present as a substituent on the alkyl chain; 5 aryl groups may be present, the aryl group being particularly preferably 1 consisting of the series nitro 1 lower alkyl (ol~04) and cX-c,-alkoxy.
They may be additionally substituted with one or two groups and these may contain up to 5 halodane atoms, preferably fluorine and chlorine.

The aryl group R1 is preferably c, -101o- or C
1ff1- is an aromatic group. These groups are preferably 0
Optionally substituted with 1-C4-alkyl, C8-C4-alkoxy or nitro, preferably with 1-2 substituents in each case or preferably with 1-5 fluorine or chlorine on the halodane atom will be replaced with

Examples of aryl and aralkyl groups R1 are phenyl, diphenyl, p-nitrophenyl, ethylphenyl1 p-methoxyphenyl, 2,4-dichlorophenyl1 benzyl1
p-methoxybenzyl~phenylethyl) phenylhexyl, tolylhegutyl 12-phenyltetradecyl and 0
1,4-phenyltetradecyl.

In the radical R1 - especially when these represent alkyl and alkenyl, and the individual methylene or methine groups may be substituted with oxygen, sulfur or nitrogen atoms, if the alkyl chain contains N in the middle, this nitrogen atom can have H or an alkyl group of up to 20 carbon atoms, preferably up to 6, or an aO-alkyl group of up to 20, preferably up to 6 carbon atoms.Generally in this process, up to 5 Preferably 1 to 5 methylene or methine groups may be substituted.

The radical R1 is preferably C56-5C10- or CI!
-Aryl, halogen 1 preferably F, C1 or Br, amino, CI to 0. -alkylamino, di0.
~C0-alkylamino, OH.

01-C0-alkoxy, o'-co-Rr or NH-00R/, R' in each case represents C1-06-alkyl, if R1 is substituted 11 or Preferably, two identical or different substituents are present.

The group R1 in formula I is located between 1 and 1! l! Cumulative, sulfur and/
Examples of a hydrocarbon group containing a nitrogen atom or substituted with a nitrogen atom include methoxyethyl, methoxyethoxyethyl, dimethylaminoethoxyethyl, digtylaminohexyl, mercaptoethyl, and oxobutyl. Aminodecyl, N-methylaminodecyl, chloroethyl 1-fluoromethyl-2-hydroxy-tetradecyl S7-phenyl octadecyl, 16-
Phenylhexadecyl 2-bromopropyl, 8-bromododecyl 116-chlorohexadecyl, 2-aminotetradecyl% 16-amituhexadecyl 14-(butylamino)hexadecyl 18-(dimethylamino)octadecyl, 8-hydroxy Octadecyl, 4-methoxydodecyl, 5-(butylcarbonyloxy)tridecyl-
2-(tridecylcarbonyloxy)tetradecyl 12
-()! J Decylcarbonylamide) Tetradecyl-2
-Hydroxyoctadec-8-Len-1-yl, 2-
Aminooctadec-8-en-1-yl and hydridecylcarbonyloxyoctadec-8-en-1-yl ~ In the group Ra, the acyl group and the alkyl group may be substituted, halogen ( F, O: L and Br, preferably 01); aryl, preferably phenyl or phenyl substituted with 1 to 2 carbon atoms or C1-C4-alkoxy group; and a group consisting of 01-0-alkoxy One or two substituents of are particularly suitable.

Examples of R1 from the acyl system are acetyl, propionyl)butyryl, pivaloyl, benzoyl, X'-methoxypency/I/%2,4-sochloropenzoyl, caproyl, myristoyl and stearoyl.

Examples of Hj from the alkyl system are methyl, ethyl 1 zorobyl 1 inglovir to henotyl, decyl, octadecyl, allyl S 1-propenyl, benzyl, p-methoxybenzyl, ethylidene, isofuropylidene, iso/thyritene 1 pene solidene, p-methoxybenzylidene and 1-methoxyethylidene.

Examples of R8 from silyl ether groups are trimethylsilyl and diethylfuropylsilyl, the phosphate group R1 is -p
o (oa), and the phosphate group Ha is the group 0, where in each case "alkyl" independently represents a carbon atom 1 to
is understood to mean 20, preferably 1 to 5 straight-chain or branched alkyl groups.

As can be seen in formula (2), the compounds according to the invention are based on sugar molecules, preferably pentoses and hekitoses.

Examples of such saccharides are ribose t-arabinose to xylose, liquinose, aldulose, glucose, mannose, gulose, pentose, thalose, and talose in both the D- and L-systems.

The compound in formula (II) has 1 or a group R as one of the groups R3.
4 always contains a phosphate group or a phosphite group, which is attached in an ester manner to the hydroxyl group of the sugar moiety.

When pentose is the sugar component of the compound in formula ■1
One of the secondary hydroxyl groups belonging to the sugar ring is phosphorylated. When a hexose forms the sugar component of the compound in formula (1), the second hydroxyl group belonging to the ring or the dorokifur group of the exocyclic hydroxymethyl group is phosphorylated.

The non-phosphorylated hydroxyl groups are either free, ie not blocked, or blocked with ether and/or silyl groups.

The compounds of formula 1 contain several chiral (ch1ra↓) atoms and are in the form of one optically pure noastereomer or diastereomer mixture.

The present invention also relates to a method for producing the compound in 2).

In this method, the sugar represented by formula IK can be used in free form, i.e. in unprotected form or in protected and optionally partially phosphorylated and/or optionally activated derivative form. Amino compound R1-HE in the acid addition salt form is first reacted with [wherein R1 has the above meaning].Next, the formula obtained by this method is 4. In one formula, RI SRa and R4 have the above meaning. is reacted with a carboxylic acid or carbonic acid derivative which is activated and optionally functionally protected in the usual manner in an acylation reaction to form a glycosylamide, glycosylcrea, glycosylcarbamate or glycosylamine. If a partially phosphorylated saccharide is not initially used, the phosphate group is introduced in a third reaction step.1 Of course, this reaction can consist of a number of reactions. splitting any protecting groups present in the reaction product;
In this way compounds of formula I are obtained and, if necessary, fi
It can be made by v.

In a preferred embodiment of the process according to the invention, unblocked sugars of the aldopentose or aldopentose type are reacted in a first step with the amines R1-NH2 in a manner known per se, with the splitting of the water occurring. te expression■
of glycosylamine. In this case ~Ra represents hydrogen and R4 represents hydrogen, methyl or hydroxymethyl.

The amine R'-NH, which is liquid at room temperature, can be reacted directly with the sugar, ie without solvent. In this case, the reaction is carried out at a temperature between 0°C and 100°C, preferably between 25°C and 70°C.
It is carried out at temperatures between ℃. Suitable catalysts are inorganic acids, such as hydrochloric acid or sulfuric acid, or short-chain carboxylic acids, such as acetic acid or noropionic acid.
Used in α05fi amount.

In all cases the amines R1-NH are solid at room temperature.

Thus, the first reaction can be carried out in the presence of a solvent, preferably a diluent which is inert under the reaction conditions and which preferably dissolves at least the reactants or reaction mixture. Performed in the presence of

Usable diluents are alcohols) such as methanol 1
Ethanol, 1-f lovanol and 2-f o ha/-
/l/, x-thyl compounds such as tetrahydrofuran and sooxane 1 and dimethylbormamide are required; addition of water is preferred if appropriate.

When a solvent is used in the production of glycosylamine, the reaction temperature is between 11oC and 120T, preferably 300C.
The temperature is between 70°C and 70°C.

The solvent in question can be added freely before or during the reaction. In the case of long-chain amineba 1-NH2, it is preferable to add it before the t-reaction.

The glycodylamines prepared as described above can be crystallized immediately or after cooling and can be hydrated by adding suitable auxiliaries, preferably low polar auxiliaries, such as acetone, diethyl ether, cyclohexane, ethyl acetate or petroleum ether. If so, it can be crystallized or crystallized by cooling. The excess amount of amine R1-N~ that is present can be removed by washing or recrystallizing the product in a manner known per se. In the second reaction step, the 4u glycosylamine obtained by this method is selectively acylated at the nitrogen atom with a carboxylic acid derivative or a carbonic acid derivative (glycosylamide).
Glycodylcarbamates, glycodylthiocarbamates or glycosylureas are produced.

When X represents a methylene group in formula 1, the corresponding glycosylamine of formula 2 is reacted with 1 to 10 equivalents of the carboxylic acid derivative of 2R", -OH!-00-Y.
R2 has the above meaning) and Y is a leaving group customary in halogeno or amidation reactions.
group), preferably an activated '6-activated x x thyl group,
Examples include p-nitrophenyl or the radical 0-Co-R, [wherein R2 has the meaning given above] or the radical 0-Co
−0−R,” [where R2 has the above meaning]
The reaction is carried out in an organic or aqueous-organic solvent, if appropriate in the presence of a base, at temperatures between 0°C and 50°C.

When X represents an oxygen atom in formula I, the glycosylamine of formula
t [where S Yti halogen, preferably chlorine, and R2 has the abovementioned meaning]. The reaction is carried out in an organic solvent, if appropriate in the presence of a base, at temperatures between -20°C and 50°C.

When X represents a sulfur atom in formula ■, the glycosylamine of formula
l-00-Y, where Y represents haOf7, preferably chlorine, and R2 has the meaning given above. The reaction is carried out in an organic solvent, if appropriate in the presence of a suitable catalyst and/or base, 1 and the reaction temperature is between 0°C and 70°C.

When X represents an NH group in the formula (2), the glycosylamine of the formula (2) is incyane) fl''-Ne.

It is reacted with 1 to 5 equivalents of [herein, R2. has the above meaning]. The reaction is preferably carried out in an organic solvent, if appropriate in the presence of a catalyst - and the reaction temperature is between -20°C and 50°C.

formula! in which X represents an Nft group or an N-alkyl group, a carbamate, a primary amine R2-BIH, or a primary amine R''-N
H-alkyl radical R4 can be reacted with 1 to 10 equivalents of 1 having the above meaning. This reaction is preferably carried out in an organic solvent, if appropriate in the presence of a catalyst, at a temperature of 20°C to 80°C.

The carboxylic acid and carbonic acid derivatives are preferably reacted with glycosylamines in the presence of a diluent that completely or only partially dissolves the reactants.

Organic or inorganic solvents can be used, preferably those which can prevent side reactions under the reaction conditions. The reaction is carried out in organic solvents such as ethers such as tetrahydrofuran or tahadioxane, alcohols such as ethanol or pro-9ol, ketones such as acetone or methyl ethyl ketone, trimethylformamide, ethyl acetate or pyridine; can be carried out in mixtures of each other and/or water. It is generally preferred to use anhydrous solvents.

The reaction with carboxylic acid derivatives or carbonate@conductors can be carried out in the presence of a basic auxiliary agent.

All basic compounds common in organic synthesis 1 such as tertiary aliphatic or aromatic amines, such as triethylamine or pyridine, or alkali metal hydroxides or carbonates or alkaline earth metal hydroxides or carbonates, such as hydroxide Sodium) Sodium carbonate or calcium carbonate can be used. The 2'H-glycosylamides, -lunogmates, -thiocarbamates or monoureas obtained in this way can be prepared in the form of a crystalline or amorphous solid or as a viscous syrup by methods known per se. isolated and optionally 1
This product is purified by crystallization 1 chromatography, extraction, etc.

In the case of compounds with a hydroxyl group protected on the glycosyl moiety, this protecting group can be cleaved off in a manner known per se.

Phosphorylation for preparing compounds of formula (1) according to the invention can be carried out by methods suitable for the synthesis of p, -oc bonds.

If one hydroxyl group is more reactive than the other hydroxyl groups in the cycle - i.e., for example the primary hydroxyl group or the 60th atom of a carbohydrate, the phosphorylation can be carried out selectively, e.g. by nucleoxide and carbohydrate chemistry. It can be carried out by conventional methods known for phosphorylation reactions. Generally such methods consist of reacting an N-alkyl-N-(fludohexobinocyl)-carboxamide, -urea, -carbamate- or -thiocarbamate with a phosphorylation reagent in an organic solvent.

Suitable solvents include a number of solvents, such as hydrocarbon beds to eg hexane, cyclohexane or toluene to halonated hydrocarbons, eg methylene chloride, chlorohexane.
cld Chlorobenzene, phenols such as phenol or cresol 1 Organic acid esters such as ethyl acetate or methyl benzoate 1 Nitro compounds such as nitromethane henitroethane or nitrobenzene 1 Nitriles such as acetonitrile followed by malononitrile 1 ethers such as diethyl Ethers such as dihydrofuran, dioxane or ethylene glycol dimethyl ether, or trialkyl acids such as trimethyl phosphate or triethyl phosphate can be used.

These solvents can be used on their own or in mixtures with each other or with organic bases such as pyridine, triethylamine or picoline or with organic bases and salts of organic or inorganic acids such as pyridine hydrochloride or pyridinium p-toluenesulfonate. It can be used as a mixture with

Suitable phosphorylation reagents include phosphorus halides (e.g., phosphorus trichloride, phosphorus oxychloride, phenylphosphorodichloridate, diphenylphosphochloridate, dipendylphospochloridate, monoalkylphosphorodichloride).
Dialkylphosphochloridate or partially hydrolyzed phosphorus oxychloride. Examples of particularly suitable combinations of reagents and solvents are phosphorus oxychloride and trialkyl or dialkyl phosphochloridates and methylene chloride/
These are toluene and pyridine.

Reaction temperature li mini -20 to +50°C, preferably 0
℃ to room temperature, and the reaction time is several hours to several days.

Either the reactive minor secondary hydroxyl group on the S residue is phosphorylated in the presence of a more reactive primary hydroxyl group, or the secondary hydroxyl group is phosphorylated in the presence of another secondary hydroxyl group. If selective phosphorylation of hydroxyl groups is not possible, the actual phosphorylation is preceded by a number of protecting group operations, and the hydroxyl groups to be phosphorylated are is selectively free, ie, exists in an unsubstituted state at the end of the blocking reaction. Therefore, hydroxyl groups that should not be phosphorylated must be blocked before phosphorylation.

Suitable protecting groups for sugar conductors are described in the relevant literature (eg OlB, R]l1ES]lI!.

Org, Ohem, 1975 (7) Protect
ingGroups, pages 95-145; pxenum
All protecting groups and combinations thereof used in PX''68 lysaccharide chemistry can be used.

Examples of suitable protecting groups are esters such as acetyl, benzoyl pivaloyl and methoxybenzoyl, ethers such as benzyl p-methoxypentyl allyl and prop-1-enyl-alkylidene compounds such as ethylidene, infropylidene. and Pe/Jiriden,
Ortho-esters, such as 1-methoxyethylidene,
1-Ethoxyethylidene, silyl ethers such as trimethylsilyl and t-butyldimethylsilyl, and organometallic compounds such as boric acid esters or tin ethers or tin ketals, such as triglythyl-stannyl or dibutyl-stannylidene.

These protecting group-blocked carbohydrate derivatives are then phosphorylated at one still free hydroxyl group in a suitable solvent. Suitable solvents and suitable methods for this phosphorylation are described above.

To prepare the free compound according to formula (1), all protective groups present are cleaved off by conventional methods.

The monopho-, sulforylated glycosylamides according to formula I, monocarbamates, 1-thiocarbamates or monoureas are isolated and purified by conventional methods. For example, adsorption chromatography on silica gel or ion exchange resins, extraction or crystallization can be used selectively or in combination.

Also included in the invention are salts of compounds of Formula I.

These salts are, inter alia, one of the customary pharmaceutically usable non-toxic salts, such as alkali metal salts, alkaline earth metal salts or ammonium salts.

The compounds of the invention exhibit useful pharmacological properties, in particular pronounced antiviral and immunostimulatory effects (1 m-munoet
imulating action).

It has been found that the compounds of the present invention inhibit viral propagation and significantly increase the potency of TFC to interfere with interferon induction by viral infection.

These effects can be demonstrated by the following experiments: Antiviral Effect Example A Animal Experiment 1 A skin test in guinea pigs was carried out by Hubler et al.

Dermatol, 62, 92-9.5, (197'4
)] performed according to the method developed by
The ventral sides of the guinea pigs were shaved at 0-600F) and anesthetized with Nembutal (15” f//h i.p.). A pre-marked area of the skin was infected with a multiscarcer (vaccine injection gun). I let it happen.

The medium of rabbit kidney cells infected with monoml virus type I was used as the viral material. Treatment can be carried out topically, parenterally, intraperitoneally or intravenously. Is it not treated 1 or Gracie? Infected animals treated with were used as controls. The plan is herpe
According to the number and size of s blisters), the compounds of the present invention are shown in Table 1.
Reduced the number and size of induced herpes.

Interferon - Stimulating Effect Example B The test was carried out by Merigan et al. [Nature-
LA'', 67 (1977)]. CF emau, x, (6-8 weeks old)
Winkelmann, Borcken) were infected intraperitoneally with a solution of lymphocytic choriomeningitis (LOM) virus 11-. A homogenized mouse brain infected with LOM was used as the viral material.

Untreated or placebo-treated infected animals served as controls.

Animals were treated prophylactically and therapeutically by intraperitoneal route (days -1, 0, 1 and 2).

Mouse serum was tested for interferon activity after the times indicated in Table 2. Before the test, the interferon-containing substance was dialyzed at a pH of 2 to inactivate any remaining virus, and the interferon level was measured using Verpel (Flavθ11).
etc. [Antimicrob, Agents C! he
mother.

E, 476 (1972)].

Table 2 Serum interferonka titer (U/ml) day 1 after treatment of LOM-infected mice with compounds in Example 6
2nd day 5th day control 0 800 2.800 5η/xt 0 5,600 4.00025■ 3,
600 9,600 6.400 Compound of the present invention - 2nd
The drugs shown in the table are able to increase the interferonka titer as a function of dosage (up to 12-fold stimulant effect).

The interferon-stimulating effect of the above-mentioned compounds is dependent on the additional stimulus (fils, interferon-inducing substance), ie, the new compound is injected together with the interferon-stimulating substance and produces an increase in interferon levels by 1 only when 7C.

As already mentioned, the compound of formula (1) according to the present invention has strong antiviral and immunostimulatory effects.

This effect applies to all viruses that are sensitive to interferon.

Examples of acute infections that can be mentioned as areas for prescription in human medicine are: viruses of the herpes group I/flu, rhinoviruses and enteric viruses.
terovirus day θF3).

persistent infection
Examples include viruses of the herpes group and hepatitis B.

Examples of prescriptions in veterinary medicine that can be cited include: pseudorabies virus (cattle and pigs), rhinotracheitis (rhino
trachitis) virus (cattle) and rhinopneumonia (
rhinopneumonxtis (horses), as well as Marek virus and chickens), Newcastle disease virus and chickens), foot and mouth diseases (cattle, pigs).

The interferon-stimulating action of the new compounds has been shown to affect, for example, macrophages and natural killer cells ("natu").
ral killer cells), making it suitable for stimulation of endogenous defenses.

Furthermore, it has been found that the compounds of the invention increase antibody synthesis of the immune system in an antigen-specific manner 1 and further strengthen endogenous non-specific defenses in the host.

These results were obtained through the following experiments.

Primary humoral immunity (r) increase against sheep red blood cells in vitro.

By primary immunization of mouse tibia cells in suspension culture, the development of a humoral immune response by atypical red blood cells can be suppressed experimentally in vitro [R, Eng., Mish.
ell and R,'W,DuttOn.

J, ExplMed, 126, 425 (1967),
I.

For this purpose, we used Ba1b/c mouse tile cells with antigen (
sm) and the test substance for 5 days.

The cells were taken out, washed once, plated on semi-solid agar with antigen and complement, and cultured at 57°C for 2 hours [1. In, Jerne, A.A.

Nor-din and C0Henryl, “Ce1l b
und Antiboiles”, p. 100 (1965
)% editors, Amoe and Kopr-owski, W.
istar engineering nst, Press, Ph1ladel
phia.

USA3. - Antigen of mouse lymphocytes in subsequent culture - Sensitization leads to the synthesis and release of antibodies (Ab), the specific antibodies released bind antigen to 13 ~ lyse these cells due to the presence of complement The substances of the invention are capable of increasing the number of antibody-producing cells as a function of dosage in the range 1-100 μm/-. Increased primary humoral immunity to the soluble antigen ovalpmin in vivo.

NMR mice were immunized subcutaneously (day c) with a suboptimal antigen dosage (1 μ2/animal, day 0). Suboptimal (suboptimal, mal) antigen stimulation stimulates only a minority of an animal's lymphocytes to antibody synthesis. By additionally treating the animal with a compound of the invention, ps50vry/
A single subcutaneous administration of ~ can significantly (p≦(Lo, 3) increase the antibody titer in the animal's serum. The antibody titer is 10
Measured by indirect blood agglutination on day 1. The effect of treatment is 1
Expressed by the geometric mean of 0g2 titers.

Other immunostimulants 1 e.g. L by Gram-negative bacteria
In contrast to bacterial immunostimulants such as PS, the immunostimulatory effect of the compounds according to the invention is antigen-dependent, i.e., surprisingly, the substances act in conjunction with the antigenic stimulus (in this case BK or ovalpmin). In contrast to common immune stimuli (eg LP8) which only lead to induction of antibody synthesis, the present compounds have no mitogenic properties.

TolerabilityCompounds of the type already mentioned can be used, for example, in 10 μ/~
Although it exhibits a strong effect in mice after multiple intraperitoneal or oral administrations, no toxic effects are observed even when 100"p/~ are administered. The above-mentioned substance is therefore well tolerated.

The compounds according to the invention have, on the one hand, the potency to increase the immunity of the antigen when mixed with the antigen and, on the other hand, the potency to increase the immunological response of the treated organism when administered systemically. ing. The substance is therefore capable of activating lymphocytes capable of responding to antibody production.

Thus, one novel compound can be used as an adjuvant in mixtures with vaccines in order to improve the outcome of vaccination and to increase the protection against infection by bacteria, viruses or immunocompromised parasitic pathogens. When mixed with the most diverse antigens as adjuvants, the compounds are also suitable for the experimental and industrial preparation of antisera for therapy and diagnosis.

Furthermore, without co-administration of the antigen, the novel compound was able to reach levels already below the threshold (θubthreshold θ).
In order to promote the defense reaction that is progressing in
Can be used in humans and animals. The compounds are therefore useful, for example, in cases of chronic and acute infections, or in cases of selective (antigen-specific) immunological deficiencies, as well as congenital and/or acquired general (i.e. antigen-non-specific) immunological deficiencies. Deficiency 1 For example, in old age, during the course of a dangerous primary disease,
and is particularly suitable for the stimulation of endogenous defenses, especially in the case of symptoms that occur after treatment with ionizing radiation or substances with immunosuppressive action. Thus, the substance preferably
It can be administered in combination with anti-infective antibiotics, chemotherapeutic agents or other healing methods to prevent immunological damage. Finally, the substance is also suitable for the general prevention of infectious diseases in humans and animals.

The compounds according to the invention can be used as prophylactic agents per se or as antibiotics and chemotherapeutic agents (1) to control existing infections.
For example, penicillin, cephalosporin 1 aminglycoside, etc.) can be used in infected humans and animals in combination with antibiotic treatments to enhance their therapeutic action.

Infection of mice with pathogenic bacteria resulting in the death of the experimental animal within 24-28 hours is treated for example by prophylactic treatment - preferably intraperitoneally - with the compounds of the invention 1-zomy/Kf as described in the Examples below. One compound that can be used to treat a large number of Durham-positive infections [e.g. Staphylococcus
taphylococci)] and goufuran negative [
For example, Escherichia coli (E. coli), Klepsiae 2 (Kl.
ebsiella), Proteus
) and Pseudomonas P4
] This reference to pathogenic fungi is for illustrative purposes only and is in no way limiting. Thus,
For example, in mice infected with the pathogenic strain Klebsiella 63,
0-100% of the bacteria survived immediately after treatment (e.g. 18 hours before infection) with the compounds 1-20~/in accordance with the invention, whereas only 0-30% of the untreated control animals survived and the bacterial serum-resistance gram Detection of phenotypic changes from negative to serum-sensitive Gram-negative strains.

Substances that modify the surface structure of Gram-negative bacteria can modify these bacteria, thus rendering them more susceptible to host defense mechanisms in the presence of inhibitory concentrations of compounds according to the invention. We found that cultured serum-resistant E. coli strains can be phenotypically converted to serum-sensitive types. This effect was demonstrated using engineered bacterial strains (C10,014, LP1674) in a total of 5 different types of E. coli.
This could be demonstrated using the compound of Example 5.

A transition occurred from serum-resistant bacteria to serum-sensitive bacteria.

Example 1 N-Octadecyl-D-glucopyranosylamine Octadecylamine 201 was dissolved in 120% of ethanol, and the solution was heated to 70°C. Anhydrous D-glucose 11f
added. After producing a clear solution, stirring was continued at 70°C for an additional 15 minutes. The solution was cooled to 10°C and left for 15 minutes. The resulting crystal sludge was washed with suction source JSushi twice with ethanol and dried under vacuum.7'Ca Elemental analysis: Calculated value: 066.8% Hli, 4% N3.2% Actual value: 0674% HI3 .8% N5.7% Example 2 Compound 102 according to Example 1 was dissolved in tetrahydrofuran 80%
After adding 10f of IJum (suspended in td and carbonated)
When one reaction in which dodecanoyl chloride 102 in tetrahydroflough 10 me was completed (toluene/
The solids were filtered off, the filtrate was evaporated under vacuum to a syrup, and the crude product was eluted on silica gel 60 in isopropanol 6:1. Purified by chromatography using a toluene/improper tool 10:1.

20.

[α), = 13 (c = = i, Q, dioxane).

Example 5 Compound 6.1f according to Example 2 was dissolved in methylene chloride 60-. To this solution was added dibenzylphosphochloridate 6f dissolved in toluene and pyridine. The batch was stirred overnight, then filtered and the filtrate evaporated under vacuum to a syrup, which was separated by column chromatography (eluent toluene/ethanol 10:1). The obtained main product (6f) was dissolved in 100 Tnt of tetrahydrofuran and 10 rJ of glacial acetic acid, and palladium (5 f) supported on charcoal was dissolved.
%) 21! Hydrogenation was carried out in the presence of.

When the hydrogen absorption was finished, the catalyst was added and the mother liquor was evaporated under vacuum to a syrup, which was co-evaporated several times with toluene.

[α]D-1,2° (c=1.0, dimethylformamide).

Ammonium salt: 20.

[α], = 8 (C = 0.065, water).

Example 4 N-dodecyl-D-galactopyranosylamine Example 1
It was prepared from D-galactose and dodecylamine according to the following.

Elemental analysis two calculated values: O/) 2.2% H1α7% N4.0% Actual value: 062.5% Ii1α2CH N4.4 Example 5 Compound 10r according to Example 4 and stearoyl chloride 16
The title compound was prepared according to Example 2 from Y.

[α] = 4.4° (Q = j, Q, methylene chloride).

Rf value = α26, toluene/n-furopanol 4:1 Example 6 Compound 6F according to Example 5, 7.5 y of dibenzylphosphochloridate and 3.9 t of Virison were then hydrogenated according to Example 5 to give the title The compound was prepared.

[α] = 11.5° (0 = a82, dimethylformamide).

Disodium salt: [α] 2po = 9° (c = α16, water).

Example 7 D-Likinose 102 was dissolved in 120 g of innolopanol and 60 g of water, and the solution was heated to 67°C. Dodecylamine 18F was added.

After producing a clear solution, this temperature was held for an additional 15 minutes. The mixture was cooled to room temperature and evaporated under vacuum at 1A. Resulting syrup + 09'/g tetrahydrofluff 10
It was dissolved in 〇- and methanol + Ome, and sodium carbonate 10F was added. Dodecanoic acid chloride 5,5F dissolved in 20 g of tetrahydrofuran was added dropwise at 0° C. When the reaction was complete, the mixture was worked up and the product was purified as described in Example 2.

[α]D = 5.5° (c = 1.1, tetrahydrofuran).

Elemental analysis: Calculated value: 069.7% Hll, 5% N2.8% Actual value: C369, 6% Hll, 7% N2.7% Example 8 Compound 5t according to Example 7 was added to tetrahydrone run 50d.
Dissolve 7'(,2,2-dimethoxypronozone 1rn
The mixture was heated to 70° C. for 50 minutes. The cold plate 1 mixture at room temperature was neutralized with ion exchange resin MP500 (OH type) and evaporated several times with toluene under vacuum. The resulting syrup was dissolved in 50 rFt of methylene chloride, 6f of dibenzylphosphochloridate and 1Tnt of pyridine were added, and the mixture was stirred overnight. The resulting solids were filtered off and the filtrate was concentrated to syrup. This syrup was purified by column chromatography (eluent: toluene/acetone 1
〇21), Dissolve the obtained main product in 50 g of methylene chloride, and add 2 m of trifluoroacetic acid/water (99:1) to 0.
Added at °C. After 10 minutes, the mixture was diluted with toluene and concentrated under vacuum. The residue was taken up in toluene and concentrated under vacuum five times.

The resulting syrup was dissolved in 80 g of tetrahydrofuran and 10 g of glacial acetic acid) and radium (5%) supported on charcoal was added.
) Hydrogenated in the presence of 2F.

When the hydrogen absorption had ended, the catalyst was filtered off and the filtrate was evaporated under vacuum to a syrup, which was co-evaporated several times with toluene.

[α]D=27 (c=α95, tetrahydrofuran)
.

Example 9 Compound 9t according to Example 1 was suspended in 160% of tetrahydrofuran and 40% of ethanol, and 9% of sodium carbonate was added.
Added t. Decyl chloroformate dissolved in tetrahydrofuran was added dropwise to this suspension over a period of 20 minutes. When the reaction was completed, the patch was filtered and the residue on the filter was rinsed with tetrahydrofuran. The filtrate was rinsed with tetrahydrofuran. and evaporated under vacuum. The resulting syrup was purified by chromatography (eluent: methylene chloride/methanol 20:1).

Rf value: α37, (3H, 01□10H, OH10:
1.

Elemental analysis: Calculated value: Ob8. ．． 5% H1t5% N2.2% Actual values: 06go 4% Hll, 6% N2.4% Example 10 Example 5 from compound 6F according to Example 9, dibenzylphosphochloridate 7,5 F and pyridine S9- The title compound was then prepared by hydrogenation according to the following procedure.

0 [α]D = 1.9° (c = α75, dimethylformamide).

Example 2 Example 11 Compound 9f according to Example 1 was dissolved in tetrahydrofuran 160
+m and 40 tneK of ethanol were suspended.

Add 20 nd! of tetrahydrofuran to this suspension. , 4.5 f of dodecyl isocyanate was added dropwise over 20 min. When the reaction was complete, the mixture was evaporated under vacuum and the resulting syrup was purified by column chromatography (eluent: methylene chloride/methanol). 15:1).

Rf I Naoji ass, 011. Cj l, 10
H, OH10: 1° [α], = 7.4° (C = 1
．． 04, dioxane).

Example 12 Compound 61 according to Example 11, dibenzylphosphochloroliso-7.5 tt and pyridine 59- to Example 3
The title compound was prepared by first hydrogenation according to the procedure.

[α] 20==t Oo (c=2.10, dimethylformamide).

Example 13 10 g of the compound according to Example 2 was dissolved in 100 Tnt of tetrahydrofuran, and benzaldehyde dimethyl acetal A5F and 1
)-) Add 10~ of luenesulfonic acid and add 1 to 7 of this mixture.
Warmed to 0°C for 1 hour. After cooling to room temperature, the mixture was neutralized with ion exchange resin MP500 (OH type) 1 and concentrated under high vacuum. The resulting syrup was poured into 10 ml of tetrahydrofuran, and after addition of 1.52 g of sodium hydride and 5 g of benzyl bromide, the mixture was heated to 40 DEG C. for 1 hour. After cooling to room temperature, 20 methanol was added and the mixture was stirred for 1 hour. 10 g of water was carefully added to the patch and the mixture was concentrated under high vacuum. The resulting syrup was taken up in 100 td of methylene chloride and extracted twice with 20 ml of sphagnum moss, dried over magnesium sulfate, concentrated in vacuo, and the residue was purified by column chromatography (eluent: toluene/acetic acid). Ethyl 2 〇 2 1)
.

The resulting syrup was dissolved in 60% of tetrahydrone, and after addition of 10:1 50% of glacial acetic acid/water, the mixture was diluted to 70%.
℃ for 1 hour, cooled to room temperature and evaporated with toluene several times under high vacuum. The resulting chromatographically homogeneous syrup (C7.8t) was dissolved in 100 μL of tetrahydrone 1. This solution was stirred with 400 μl of sodium hydride at 50° C. for 50 minutes and cooled to SO0°)) benzyl bromide. Added 8f. After 5 hours, the mixture was humidified to room temperature and stirred overnight. After addition of 5 g of methanol, the mixture was stirred for 1 hour, then water was carefully added and the mixture was evaporated. The resulting residue was taken up in methylene chloride and water, and the organic phase was extracted with water, dried and concentrated under high vacuum.

The residue was taken up in a 10-liter tetrahydrone column and the solution was stirred overnight with 5.5 F of dibenzylphosphochloroliso-) and 2.8 Tnt of pyridine. The mixture was filtered and the filtrate was evaporated under vacuum to a syrup which was purified by chromatography (
Dissolved Ed agent 2) Luene/ethyl acetate 15:1).

The main product obtained (5,9F) was hydrogenated in the presence of 2F of palladium on charcoal, dissolved in 80g of tetrahydrone and 10g of glacial acetic acid. When the absorption of hydrogen had ended, one catalyst was filtered off and one filtrate was evaporated under vacuum to a syrup, which was evaporated several times with toluene.

[α], , = 1.9° (C = 2.0, dimethylpolymer, V mid).

In a manner similar to Examples S and 6, the aldose was reacted with an alkylamine to form an aldosylamine, acylated with an aliphatic chloride to form an aldosylamide, and then selectively phosphorylated and The following compounds were synthesized by hydrogenation: Example 14 Mid 0 [α]D=12° (C=1.24, dimethylformamide) Example 15 Mid Example 16 Acid amide Example 17 2-nosyl-6 -phosphate)-oleic acid amide Example 18 Acid amide example 19 Acid amide example 20 Mid Example 21 Dinosyl-6-phosphate)-dodecanoic acid amide 20 disodium salt: [α]o=-0 ,1゜(Q=145,
water) Example 22 Acid amide Example 23 Acid amide example 24 Amide [α] , =-1° (c=055, water) Example 25 Amide disodium salt: [α]2D. = + 4.0゜(c=
α1, water) Example 26 Example 27 Amide disodium salt: [α]2D. -16° (c=α7, water) Example 28 Amide disodium salt: [α]20=+40 (a=α25, water) Example 29 Amide example 50 Amide practical example 51 N-(dodecyl)-N -'(D-mannopyranosyl-6
- Phosphate) - Octadecanoic acid amide Continued from page 1 Priority claim [Phase] 1% rhinoceros February 2 [Phase] West Germany @
Inventor Beter Stadler@Inventor Arnold Perasentos Yu@Inventor Gerd Schudreis Dorezi@Inventor Beter Fist Tyler Ack@Inventor Hansjoachim Zdeiler - 0 Inventor Lurugeorg Mel Drager Yu (D E) @P3403495.1 Day of the Federal Republic of Italy 5657 Hartham Ideck 8
Federal Republic of Germany Day 5657 Hartschtresemansittrasse 56 Federal Republic of Germany Day 5600 Wuppertalt am Hohotstu 17 West Haven Mobinrein, Connecticut, United States 06516 400. Molecular Fist Diagnostics Incorporated Day 5620 Felbert 15 Elsbekan Uttrasse 46 Day 5600 Wuppertaltbarke Sittrasse 75

Claims (1)

[Claims] 1. In the general formula, R1 is a linear or branched saturated 1-unsaturated or polyunsaturated alkyl group which may be optionally substituted;
The optionally substituted saturated, monounsaturated or polyunsaturated cycloalkyl group represents an optionally substituted aryl group or aralkyl group, XhaOH2, ○, 8. If it represents NH or N-alkyl of up to 20 carbon atoms and R2 has the meaning of R1 and X represents OH, it can represent ~hydrogen, the radicals Hj being mutually independently hydrogen, in each case an acyl or alkyl group of up to 20 carbon atoms, a silyl group and/or a phosphoric acid or phosphoric ester group) and R4 represents hydrogen to methyl or 10H2-ORB; one of which always represents a phosphoric acid or phosphate ester group. 2. A compound according to claim 1, in which R1 represents an alkyl group or an alkenyl group having 10 to 20 carbon atoms. & Compounds according to claim 1 or 2, wherein the alkyl or acyl group R3 has 1 to 10 carbon atoms; 4. The phosphoric acid group is a group of the formula -PO(OH); Any one of claims 1 to 5, wherein the acid ester group is a group of the formula %, where hairkyl represents a linear or branched alkyl group having 1 to 20 carbon atoms. The compound described in 5. The alkyl group of the phosphoric acid ester group has 1 to 5 carbon atoms.
5. The compound according to claim 4, having: & Pentose or hexose 2R1-NH! [In the formula, Rt has the meaning described in claim 1]
reacted with an amine of the formula ■ 4 in which Ha and R4 have the meaning given in claim 1 - provided that one of the R3 groups must not represent a phosphoric acid or phosphate ester group; 1 The compound of the formula
00-X -R2 [where R2 is the first claim
acylation against the insertion of [with the meaning given in Section 1] and, if appropriate, followed by phosphorylation, if one of the radicals R8 in formula (2) does not already represent a phosphoric acid or phosphate ester group. A method for producing the compound according to claim 1, characterized in that: 1. A drug containing at least one compound according to any one of claims 1 to 5. a. Use of the compound according to claim 1 in the control of diseases. Claim 1 for controlling viral diseases
Use of a compound according to any of items 5 to 5. 1α Use of a compound according to any of claims 1 to 5 for immunostimulatory action. 11. Use of a compound according to any of claims 1 to 5 in controlling bacterial infections.