Amphiphilic compound having dendronic hydrophobic group and application thereof

Patent metadata

Field	Value
Publication number	US-11524975-B2
Application number	US-201816755580-A
Country	US
Kind code	B2
Filing date	Feb 28, 2018
Priority date	Oct 11, 2017
Publication date	Dec 13, 2022
Grant date	Dec 13, 2022

Abstract

The present invention relates to an amphiphilic compound having a dendronic hydrophobic group, a method for preparing the same, and a method for extraction, solubilization, stabilization, or crystallization of a membrane protein by using the same. The use of the compound according to the present invention leads to an excellent membrane protein solubilization effect and a stable storage of a membrane protein in an aqueous solution for a long time, and thus can be utilized for functional analysis and structural analysis of the membrane protein. Especially, the amphiphilic compound having a dendronic hydrophobic group showed very remarkable characteristics in the visualization of protein composites through an electronic microscope. The membrane protein structural and functional analysis is one of the fields of greatest interest in current biology and chemistry, and more than half of the new drugs that are currently being developed are targeted at membrane proteins, and thus the amphiphilic compound of the present invention can be applied to membrane protein structure studies closely related to the development of new drugs.

First claim

The invention claimed is:

1. A compound represented by Formula 1 below:

wherein R 1 to R 4 are each independently a substituted or unsubstituted C 1 -C 15 alkyl group, a substituted or unsubstituted C 1 -C 15 cycloalkyl group, or a substituted or unsubstituted C 1 -C 15 aryl group; A 1 to A 4 are each independently —CH 2 —, oxygen (O) or sulfur (S); and X 1 to X 3 are each independently an oxygen-linked saccharide.

2. The compound of claim 1 , wherein the saccharide is a monosaccharide or disaccharide.

3. The compound of claim 1 , wherein the saccharide is glucose or maltose.

4. The compound of claim 1 , wherein R 1 to R 4 are each independently a substituted or unsubstituted C 1 -C 10 alkyl group; A 1 to A 4 are each independently oxygen (O) or sulfur (S); and X 1 to X 3 are each independently glucose or maltose.

5. The compound of claim 1 , wherein R 1 to R 4 are each independently a substituted or unsubstituted C 1 -C 10 alkyl group; A 1 to A 4 are —CH 2 —; and X 1 to X 3 are glucose or maltose.

6. The compound of claim 1 , wherein the compound is one of Formulas 2 to 8 below:

7. The compound of claim 1 , wherein the compound is an amphiphilic molecule for extracting, solubilizing, stabilizing, crystallizing or analyzing a membrane protein.

8. The compound of claim 1 , wherein the compound has a critical micelle concentration (CMC) of 0.0001 to 1 mM in an aqueous solution.

9. A composition for extracting, solubilizing, stabilizing, crystallizing or analyzing a membrane protein, comprising the compound of claim 1 .

10. The composition of claim 9 , wherein the composition is a formulation in the form of a micelle, liposome, emulsion or nanoparticle.

11. A method of preparing a compound represented by Formula 1, comprising:
1) synthesizing a dialkylated mono-ol derivative by introducing an alkyl group to dimethylmalonate and performing reduction;
2) synthesizing tetra-alkylated methallyl diether to which four alkyl groups are introduced by adding methallyl dichloride to the product of Step 1);
3) synthesizing a tetra-alkylated tri-ol derivative by reacting 4-(bromomethyl)-methyl-2,6,7-trioxabicyclo[2,2,2]-octane with the product of Step 2);
4) introducing a protecting group-attached saccharide by performing maltosylation on the product of Step 3); and
5) performing deprotection on the product of Step 4):

wherein R 1 to R 4 are each independently a substituted or unsubstituted C 1 -C 15 alkyl group, a substituted or unsubstituted C 1 -C 15 cycloalkyl group, or a substituted or unsubstituted C 1 -C 15 aryl group; A 1 to A 4 are —CH 2 —; and X 1 to X 3 are each independently an oxygen-linked saccharide.

12. The method of claim 11 , wherein R 1 to R 4 are each independently a substituted or unsubstituted C 1 -C 10 alkyl group; and X 1 to X 3 are maltose.

13. A method of preparing a compound represented by Formula 1 below, comprising:
1) synthesizing a dialkylated mono-ol derivative (ether-functionalized dialkylated mono-ol derivative) by reacting an aliphatic alcohol or alkylthiol with methallyl dichloride;
2) synthesizing a tetra-alkylated mono-ol derivative (ether-functionalized tetra-alkylated mono-ol derivative) by reacting methallyl dichloride with the product of Step 1);
3) synthesizing a tetra-alkylated tri-ol derivative by reacting 4-(bromomethyl)-methyl-2,6,7-trioxabicyclo[2,2,2]-octane with the product of Step 2);
4) introducing a protecting group-attached saccharide by performing maltosylation on the product of Step 3); and
5) performing deprotection on the product of Step 4):

wherein R 1 to R 4 are each independently a substituted or unsubstituted C 1 -C 15 alkyl group, a substituted or unsubstituted C 1 -C 15 cycloalkyl group, or a substituted or unsubstituted C 1 -C 15 aryl group; A 1 to A 4 are each independently oxygen (O) or sulfur (S); and X 1 to X 3 are an oxygen-linked saccharide.

14. The method of claim 13 , wherein R 1 to R 4 are each independently a substituted or unsubstituted C 1 -C 10 alkyl group; and X 1 to X 3 are maltose.

15. A method of extracting, solubilizing, stabilizing, crystallizing or analyzing a membrane protein, comprising treating a membrane protein with a compound represented by Formula 1 below in an aqueous solution:

wherein R 1 to R 4 are each independently a substituted or unsubstituted C 1 -C 15 alkyl group, a substituted or unsubstituted C 1 -C 15 cycloalkyl group, or a substituted or unsubstituted C 1 -C 15 aryl group; A 1 to A 4 are each independently —CH 2 —, oxygen (O) or sulfur (S); and X 1 to X 3 are an oxygen-linked saccharide.

16. The method of claim 1 , wherein R 1 to R 4 are each independently a substituted or unsubstituted C 1 -C 10 alkyl group; A 1 to A 4 are each independently oxygen (O) or sulfur (8); and X 1 to X 3 are each independently glucose or maltose.

17. The method of claim 15 , wherein R 1 to R 4 are each independently a substituted or unsubstituted C 1 -C 10 alkyl group; A 1 to A 4 are —CH 2 —; and X 1 to X 3 are each independently glucose or maltose.

18. The method of claim 15 , wherein the membrane protein is a light harvesting-I and reaction center complex (LII-R complex), a leucine transporter (LeuT), a human β 2 adrenergic receptor (β 2 AR), melibiose permease (MelB), or a combination of two or more thereof.

Assignees

• Iucf Hyu Erica Campus

Inventors

• Chae Pil Seok
• Sadaf Aiman

Classifications

• G01N1/28

  Preparing specimens for investigation {including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q}(mounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 {; laboratory gas handling apparatus B01L5/00}) · CPC title

• G01N33/6842

  Proteomic analysis of subsets of protein mixtures with reduced complexity, e.g. membrane proteins, phosphoproteins, organelle proteins · CPC title

• C07H15/04 Primary

  attached to an oxygen atom of the saccharide radical · CPC title

• G01N33/68

  involving proteins, peptides or amino acids {(involving lipoproteins G01N33/92)} · CPC title

• C07K1/145

  by extraction or solubilisation · CPC title