Synthetic glycolipides, a process for the preparation thereof and several uses for these synthetic glycolipides

• US 4,751,219 A
• Filed: 02/05/1985
• Issued: 06/14/1988
• Est. Priority Date: 02/05/1985
• Status: Expired due to Fees

• Current Assignee: NEDERLANDSE CENTRALE ORGANISATIE VOOR TOEGEPAST-NATUUR-WETENSCHAPPELIJK ONDERZOEK, JULIANA VAN STOLBERGLAAN 148, 2595 CL THE HAGUE, THE NETHERLANDS
• Original Assignee: Nederlandse Centrale Organisatie Voor Toegepast-Natuur-Wetenschappelijk Onderzoek
• Inventors: Kempen, Hermanus J. M.
• Primary Examiner(s): Griffin, Ronald W.
• Application Number:
  US06/698,325
• Time in Patent Office: 1,225 Days
• Field of Search: 514/26, 536/5, 424/450, 424/461
• US Class Current: 514/26
• CPC Class Codes: A61K 47/26 Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
• CPC Class Codes: A61K 47/28 Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/1272 with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids with cholesterol as the only non-phosphatidyl surfactant A61K9/127; lipids as modifying agent A61K47/543
• CPC Class Codes: C07H 13/06 Fatty acids
• CPC Class Codes: C07H 15/04 attached to an oxygen atom of the saccharide radical
• CPC Class Codes: C07J 41/0055 the 17-beta position being substituted by an uninterrupted chain of at least three carbon atoms which may or may not be branched, e.g. cholane or cholestane derivatives, optionally cyclised, e.g. 17-beta-phenyl or 17-beta-furyl derivatives
• CPC Class Codes: C07J 43/003 not condensed

Abstract

Synthetic glycolipides having the general formula lipidyl-O-R wherein the lipidyl moiety is selected from the group consisting of 5-cholestene-3β-yl, ceramidyl, phosphatidylethanolamine and diacylglycerol and R represents a group having the formula ##STR1## in which two symbols Z represent a glycopyranosyl rest and the other symbol Z a hydrogen atom or all three symbols Z represent a glycopyranosyl rest. These new glycolipides can be used for the solubilization of cholesterol in water or aqueous media, for the manufacture of lipide-vesicles or lipoproteins in which such a glycolipide is encapsulated as well as for the reduction of the cholesterol level in the blood plasma of mammals. Preferably the compound N-tris(β-D-galactopyranosyloxymethyl)methyl-Nα-(4-(5 cholesten-3β-yloxy)succinyl)glycinamide is used.

Claims

1. Synthetic glycolipides having the general formula lipidyl-O-R, in which the lipidyl moiety is selected from the group consisting of 5-cholesten-3-β

-yl, ceramidyl, phosphatidylethanolamine and diacylglyceryl and R represents a group having the formula ##STR10## wherein two symbols Z represent a glycopyranosyl rest and the other symbol Z is a hydrogen atom or all three symbols Z represent a glycopyranosyl rest.

2. Glycolipide according to claim 1, characterized in that the glycolipide is N-tris-(β

-D-galactopyranosyloxymethyl)methyl-N.sup.α

(4-(5-cholesten-3β

-yloxy)succinyl)glycinamide.

3. Glycolipide according to claim 1, characterized in that the glycolipide is N-tris-(β

-D-2-acetamido-2-deoxygalactopyranosyloxymethyl)methyl-N.sup.α

(4-(5-cholesten-3β

-yloxy)succinylglycinamide.

4. A process for the preparation of glycolipides according to claim 1, characterized by the following stages:

(a) N-(2-hydroxy-(1,1-dihydroxymethyl)-ethyl)-N.sup.α

-benzyloxycarbonylglycinamide is prepared by reacting 2-amino-2-hydroxymethyl-1,3-propanediol with benzyloxycarbonylglycine in the presence of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroxyquinoline;

(b) per-O-acetyl-D-glycopyranosylbromide is prepared by reacting a per-O-acetyl-D-glycopyranose with PBr₃ in acetic anhydride;

(c) N-(tris(per-O-acetyl-D-glycopyranosyloxymethyl)methyl)-N.sup.α

-benzyloxycarbonylglycinamide is prepared by reacting N-(2-hydroxy-(1,1-dihydroxymethyl)-ethyl)N.sup.α

-benzyloxycarbonylglycin amide, kept in suspension in CH₃ CN with Hg(CN)₂ and HgBr₂ with per-O-acetyl-D-glycopyranosylbromide, dissolved in CH₃ CN and purifying the product by column chromatography;

(d) N-(tris(per-O-acetyl-D-glycopyranosyloxymethyl)methyl)-N.sup.α

-benzyloxy carbonylglycinamide is converted into N-(tris-glycopyranosyloxymethyl)methyl)glycine.HCl-salt by treating the compound with subsequently triethanolamine, acetic acid, hydrogen and palladium/carbon and HCl after which the compound is purified by gel filtration;

(e) one of the lipides selected from the group consisting of cholesterol, ceramide, phosphatidylethanolamine and diacylglycerol is reacted with succinic acid in pyridine into the corresponding hemisuccinyl ester and this ester is then converted into the N-hydroxy-succinimidyl-4-(lipidyl)succinate by reacting said ester with N-hydroxy-succinimide in tetrahydrofuran in the presence of dicyclohexylcarbodiimide;

(f) N-(tris(glycopyranosyloxymethyl)methyl-N.sup.α

-(4-lipidyl)succinylglycinamide is prepared by combining N-(tris(glycopyranosyloxymethyl)methyl)glycine.HCl dissolved with N-ethylmorpholine in dimethylformamide, with N-hydroxy-succinimidyl-4-(lipidyl)succinate, dissolved in dimethylacetamide;

(g) N-tris(glycopyranosyloxymethyl)methyl-N.sup.α

-(4-lipidyl)succinylglycinamide according to the invention as well as the corresponding bis(glycopyranosyloxymethyl)-derivative are purified by column chromatography.

5. A process for the preparation of N-tris-(β

-D-galactopyranosyloxymethyl)methyl-N.sup.α

(4-(5-cholesten-3β

-yloxy)succinyl)glycinamide characterized by carrying out the process as described in claim 4 using as starting compounds 5-cholesten-3β

-ol as lipide in stage e) and penta-O-acetyl-β

-D-galactopyranose as acylated monohexose in stage b).

6. Lipide vesicles or lipoproteins having encapsulated therein a glycolipide according to claim 1 characterized by dissolving the glycolipide according to claim 1 in water or aqueous salt solution and mixing this solution with a suspension of lipide vesicles or lipoproteins in water or aqueous salt solution.

7. Lipide vesicles or lipoproteins according to claim 6 characterized in that the glycolipide is N-tris-(β

-D-galactopyranosyloxymethyl)methyl-N(4-(5-cholesten-3β

-yloxy)succinyl)gylcinamide.

8. A process for solubilizing cholesterol in water or aqueous media characterized by mixing a solution of cholesterol in ethanol with a solution of a glycolipide according to claim 1 in water.

9. A process according to claim 8 characterized in that a solution of cholesterol and ethanol is mixed with a solution of N-tris-(β

-D-galactopyranosyloxmethyl)methyl-N(4-(5-cholesten-3β

-yloxy)succinyl)glycinamide.

10. A process for reducing the cholesterol level in blood plasma of mammals characterized by administering thereto a glycolipide according to claim 1 as solution in an aqueous salt solution by intravenous infusion.

11. A process according to claim 10 characterized in that N-tris-β

-D-galactopyranosyloxmethyl)methyl-N(4-(5-cholesten-3β

-yloxy)succinyl)gylcinamide.