JP6341479B2 - Pseudocyclic lipid compounds - Google Patents

Description

  The present invention relates to a novel pseudo-cyclic lipid compound.

  Lipids that have a structure in which one molecule of glycerol and two molecules of saturated or unsaturated fatty acid are ester-bonded have surface activity, and are used as various industrial treatment agents for mining, metal processing, surface finishing, and cleaning. It is widely used as various household detergents and detergents, pharmaceuticals, cosmetics and food additives. Among these uses, it is particularly useful as an emulsifier, demulsifier, detergent, dispersant, hydrotrope and the like.

  Among glycerol esters of saturated or unsaturated fatty acids of this type, two molecules of saturated or unsaturated fatty acids are ester-bonded to two hydroxyl groups of one molecule of glycerol, and phosphoric acid groups are bonded to the remaining hydroxyl groups. Etc. exist in the living body and have been synthesized by artificial means.

  In order to improve the properties of the glycerol ester compound for application to the above-mentioned uses, various studies have been made, and those having a cyclic moiety in the molecule have been synthesized. Such compounds have attracted attention because the characteristics of molecular rigidity are extremely improved, but there are problems such as difficulty in synthesis and excessively high rigidity of the compounds (Non-Patent Documents 1 to 3). Then, although the cyclic type compound which introduce | transduced the methyl branched chain in order to supplement a softness | flexibility was reported, the synthesis | combination was also difficult (nonpatent literature 4). On the other hand, quasi-cyclic compounds having moderate flexibility and easy synthesis have been reported (Non-Patent Documents 3 to 15).

  Fluorine-containing compounds are useful as ingredients of pharmaceuticals, agricultural chemicals, etc., or intermediates thereof, and use their surface-active action, for example, in the fields of metal processing, mining, surface finishing, cleaning and cleaning, and cooking. , Emulsifiers, demulsifiers, detergents, dispersants and wetting agents can be used for various industrial and household applications.

  For example, fluorosurfactants represented by perfluorooctane sulfonic acid (PFOS) and perfluorooctanoic acid (PFOA) in which a sulfo group or a carboxyl group is introduced into a fluorinated alkyl group are well known. Compared to surfactants and the like, it has excellent properties such as surface tension reducing ability (Non-Patent Documents 16 and 17).

Bull. Chem. Soc. Jpn., 70, 2545 (1997) Bull. Chem. Soc. Jpn., 72, 1575 (1999) Langmuir, 16, 10340 (2000) J. Org. Chem., 63, 2689 (1998) J. Am. Chem. Soc., 112, 3188 (1990) Langmuir, 7, 2415 (1991) Langmuir, 8, 637 (1992) J. Org. Chem., 57, 1777 (1992) J. Am. Chem. Soc., 115, 3458 (1993) J. Biochem, 119, 115 (1996) J. Org. Chem., 61, 7382 (1996) Tetrahedron Lett. 41, 2971 (2000) Langmuir, 16, 128 (2000) Chem. Eur. J., 8, 585 (2002) Curr. Opin. Colloid. Interface Sci., 8, 466 (2004) E. Kissa, “Fluorinated Surfactants and Repellents”, 2nd ed., Marcel Dekker, New York (2001) Curr. Opin. Colloid Interf. Sci., 8, 227 (2003)

  So far, no quasi-cyclic compound having a fluorine atom or a non-covalent acetylene group introduced at the non-terminal has been known. An object of the present invention is to provide a novel glycerol ester compound or a glycerol ether compound, in which a fluorine atom and / or a non-terminal triple bond or both are introduced into the molecule, which is relatively easy to synthesize, and a molecular assembly containing them Is to provide.

  The inventors of the present invention have made extensive studies on the above problems, and are novel quasicyclic compounds in which two molecules of glycerol are bound to monovalent and divalent alcohols or carboxylic acids via ether or ester bonds, and fluorine molecules are contained in the molecules. We have successfully synthesized atoms or non-terminal triple bonds or both. More specifically, by reacting a glycerol derivative with an alcohol or carboxylic acid or a derivative thereof, a quasicyclic compound formed by four ester or ether bonds is synthesized, and a fluorine atom or a non-terminal compound is formed in the molecule. Triple bonds or both were introduced. Furthermore, by using chiral glycerol derivatives as raw materials, we found that a new group of chiral quasi-cyclic compounds can be selectively produced, created molecular assemblies containing them, and completed the present invention. I let you.

  A chiral glycerol derivative is a non-reactive state by protecting the hydroxyl group at the 1-position, 2-position or 1-position and 2-position of glycerol, obtained by optical resolution or resolution using an enzyme, or (S)-(+)-2,2-dimethyl-1,3-dioxolane-4-methanol or (R)-(−)-2,2-dimethyl-1,3-dioxolane-4-methanol, or ( R)-(+)-3-benzyloxy-1,2-propanediol or (S)-(−)-3-benzyloxy-1,2-propanediol is used for the reaction.

［式中、Ｒ^１は２価の炭素数１以上１００以下の有機基、Ｒ^２およびＲ^３は１価の炭素数１以上１００以下の有機基を表し、Ｒ^１〜Ｒ^３のいずれか１以上にフッ素原子を１個以上有する、もしくは非末端に非共役の三重結合を１個以上有する、もしくはその両者を有する。また、Ｒ^２とＲ^３は同一であっても、異なるものであっても良い。Ｘ^１〜Ｘ^４はそれぞれ独立してメチレン基もしくはカルボニル基、Ｙ^１およびＹ^２はそれぞれ独立して水素原子、リン酸、ホスホコリン、ホスホセリン、ホスホエタノールアミンもしくはホスホグリセロールを表す。グリセロール部位の立体構造は、（Ｒ、Ｒ）（Ｓ、Ｓ）（Ｒ、Ｓ）（Ｓ、Ｒ）もしくはこれらの混合物で表される。］
［２］上記［１］記載の擬環状化合物から構成される分子集合体、もしくは該擬環状化合物およびそれと異なる１種以上の脂質から構成される分子集合体。
［３］更にタンパク質を含む、上記［２］記載の分子集合体。
［４］上記［１］記載の擬環状化合物を構成成分とするリポソーム。 That is, the present invention specifically provides the following inventions.
[1] Pseudocyclic compounds represented by the following general formulas (1) to (3).

[Wherein, R ¹ represents a divalent organic group having 1 to 100 carbon atoms, R ² and R ³ represent a monovalent organic group having 1 to 100 carbon atoms, and any one of R ^{1 to} R ³ As described above, it has one or more fluorine atoms, or one or more non-conjugated triple bonds at the non-terminal, or both. R ² and R ³ may be the same or different. X ^{1 to} X ⁴ each independently represents a methylene group or a carbonyl group, and Y ¹ and Y ² each independently represent a hydrogen atom, phosphoric acid, phosphocholine, phosphoserine, phosphoethanolamine or phosphoglycerol. The steric structure of the glycerol moiety is represented by (R, R) (S, S) (R, S) (S, R) or a mixture thereof. ]
[2] A molecular assembly composed of the quasi-cyclic compound according to [1] above, or a molecular assembly composed of the quasi-cyclic compound and one or more different lipids.
[3] The molecular assembly according to the above [2], further comprising a protein.
[4] A liposome comprising the pseudo-cyclic compound according to [1] above as a constituent component.

  According to the present invention, a pseudo-cyclic compound having two glycerols and four ester or ether bonds in the same molecule can be obtained. These compounds are surface-active, especially in the fields of metalworking, mining, surface finishing, cleaning and cleaning, cosmetics, medicine and food processing and cooking in industry and household, especially emulsifiers, demulsifiers, cleaning agents. , Dispersants and hydrotropes. Further, it can form a molecular assembly from a pseudo-cyclic compound or a different compound and can be used as a membrane protein reconstituted membrane lipid.

2 shows the CD spectrum of compound 13 of the present invention. 1 shows the CD spectrum of compound 36.

  The novel pseudocyclic lipid compound of the present invention can be represented by the following general formulas (1) to (3).

［式中、Ｒ^１は２価の炭素数１以上１００以下の有機基であり、Ｒ^２およびＲ^３は１価の炭素数１以上１００以下の有機基を表し、Ｒ^１〜Ｒ^３のいずれか１以上にフッ素原子を１個以上有する、もしくは非末端に非共役の三重結合を１個以上有する、もしくは両者を有する。また、Ｒ^２とＲ^３は同一であっても、異なるものであっても良い。Ｘ^１〜Ｘ^４はそれぞれ独立してメチレン基もしくはカルボニル基、Ｙ^１およびＹ^２はそれぞれ独立して水素原子、リン酸、ホスホコリン、ホスホセリン、ホスホエタノールアミンもしくはホスホグリセロールを表す。グリセロール部位の立体構造は、（Ｒ、Ｒ）（Ｓ、Ｓ）（Ｒ、Ｓ）（Ｓ、Ｒ）もしくはこれらの混合物で表される。］
上記の式中の有機基Ｒ^１からＲ^３に含まれる炭素数は、１以上、好ましくは７以上、さらに好ましくは１２以上であり、１００以下、好ましくは５０以下、さらに好ましくは３６以下である。

[Wherein, R ¹ represents a divalent organic group having 1 to 100 carbon atoms, R ² and R ³ represent a monovalent organic group having 1 to 100 carbon atoms, and any ^one of R ^{1 to} R ³ Or having at least one fluorine atom at one or more, or having at least one non-conjugated triple bond at the non-terminal, or both. R ² and R ³ may be the same or different. X ^{1 to} X ⁴ each independently represents a methylene group or a carbonyl group, and Y ¹ and Y ² each independently represent a hydrogen atom, phosphoric acid, phosphocholine, phosphoserine, phosphoethanolamine or phosphoglycerol. The steric structure of the glycerol moiety is represented by (R, R) (S, S) (R, S) (S, R) or a mixture thereof. ]
The number of carbon atoms contained in the organic groups R ¹ to R ³ in the above formula is 1 or more, preferably 7 or more, more preferably 12 or more, 100 or less, preferably 50 or less, more preferably 36 or less. .

R ¹ to R ³ each have one or more fluorine atoms, or one or more non-conjugated triple bonds at the non-terminal as unsaturated, or both. Preferably, any of R ¹ to R ³ includes a perfluoroalkyl group having 1 to 18 carbon atoms. The number of triple bonds is preferably 1 or 2. R ¹ to R ³ may include a conjugated triple bond in addition to the above.

The organic group (monovalent organic group) constituting R ² and R ³ includes an aliphatic group, an aromatic group, and a heterocyclic group.

  Aliphatic groups include chain and cyclic groups, as well as saturated and unsaturated groups. The chain aliphatic group includes an alkyl group and an alkenyl group. Cycloaliphatic groups include cycloalkyl groups and cycloalkenyl groups.

  In the alkyl group, the number of carbon atoms constituting the main chain is preferably 1 to 50, more preferably 1 to 36.

  In the alkenyl group, the number of carbon atoms constituting the main chain is preferably 2 to 50, more preferably 2 to 36. In the cycloalkyl group and the cycloalkenyl group, the number of rings can be one or more (2 to 4, preferably 2 to 3), and the number of carbon atoms constituting all the carbocycles contained in the molecule is 5 -60, preferably 6-36.

  The aromatic group includes a monocyclic group and a polycyclic group, and the polycyclic group includes a condensed polycyclic group and a chain polycyclic group. More specifically, the aromatic group includes an aryl group and an aralkyl group.

  The aryl group can have a monocyclic or polycyclic structure, and the number of carbon atoms constituting the total number of carbon atoms contained in the molecule is 6 to 60, preferably 6 to 36. The aralkyl group can have a monocyclic or polycyclic structure, and the number of carbon atoms constituting all carbocycles contained in the molecule is 7 to 70, preferably 7 to 36.

  The heterocyclic group includes an aliphatic heterocyclic group and an aromatic heterocyclic group. The ring constituent elements constituting the heterocyclic group include one or more hetero elements (oxygen, nitrogen, sulfur, selenium, etc.).

  The heterocyclic group can have a monocyclic or polycyclic structure, and the number of elements constituting the entire heterocyclic ring contained in the molecule is 5 to 50, preferably 5 to 36.

  Aromatic heterocyclic groups include thiophene ring, furan ring, pyrrole ring, pyridine ring, quinoxaline ring, purine ring, oxazole ring, benzoxazole ring, naphthoxazole ring, thiazole ring, benzothiazole ring, naphthothiazole ring, selenazole ring Benzoselenazole ring, naphthoselenazole ring, imidazole ring, benzimidazole ring, naphthimidazole ring, quinoline ring, quinoxaline ring, purine ring, acridine ring, phenanthroline ring, etc. .

  Examples of the aliphatic heterocyclic group include those derived from an aliphatic heterocyclic ring such as a pyrazoline ring, a pyridine ring, a piperidine ring, an indoline ring, a morpholine ring, a pyran ring, an imidazolidine ring, a thiazoline ring, an imidazoline ring, and an oxazoline ring. Can do.

  Specific examples of the organic group include, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, isopentyl, 2-methylbutyl, 1-methylbutyl, n -Hexyl, isohexyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, heptyl, octyl, isooctyl, 2-ethylhexyl, nonyl, decyl, undecyl, dodecyl, tetradecyl, hexadecyl, octadecyl, eicosyl, cyclopentyl, cyclo Hexyl, adamantyl, vinyl, propenyl, butenyl, acrylic, methacrylic, octynyl, dodecenyl, undecenyl, cyclohexenyl, phenyl, tolyl, pyrenyl, phenanthrenyl, naphthyl, biphenylyl, terphenylyl, benzyl, vinyl Nechiru, other naphthylmethyl, and a heterocyclic group derived from various heterocycles mentioned above.

The divalent organic group constituting R ¹ includes an organic group obtained by removing one hydrogen atom from various monovalent organic groups shown for R ² and R ³ .

The organic group constituting R ¹ to R ³ may have an inert substituent that does not participate in the synthesis reaction of the present invention. Such substituents include substituted or unsubstituted aryl groups, carbonyl groups, alkoxy groups, alkoxycarbonyl groups, acyl groups, acyloxy groups, alkyl or arylsulfonyl groups, nitro groups, halogens (fluorine atoms, chlorine atoms, bromine atoms). , Iodine atoms) and the like.

In addition, the organic groups R ¹ to R ³ may contain a different atom (hetero atom) in the main chain of the organic group. The heteroatom includes an oxygen atom, a nitrogen atom, a sulfur atom, a silicon atom and the like.

Further, the organic groups R ¹ to R ³ may contain a metal atom. In this case, the metal atoms include alkali metals such as lithium, sodium, potassium, rubidium and cesium, alkaline earth metals such as beryllium, magnesium, calcium, strontium and barium, metals such as boron, aluminum, titanium, tin and iron Atoms can be mentioned. These metal atoms are usually contained in the organic group in a bond form such as a carbon-metal bond or an ionic bond.

The organic groups R ¹ to R ³ in the present invention may be a sugar compound residue (a group obtained by removing one hydrogen atom from a sugar compound). The sugar compounds in this case include monosaccharides, amino sugars, oligosaccharides, and modified products thereof.

The monosaccharide compound has the following general formula (4)
C _n (H ₂ O) _n (4)
[Wherein, n is a number of 1 to 10, preferably 5 to 8. ]
It is a compound represented by these. This monosaccharide compound preferably includes pentose, hexose, deoxyhexose, heptose and the like.

  Specific examples of monosaccharide compounds include, for example, arabinose, ribose, xylose, glucose, galactose, mannose, fructose, rhamnose, fucose, digitoxose, timarose, oleandrose, digitalose, abiose, hamamelose, streptose, cedoheptulose, coliose Etc.

  Specific examples of amino sugar compounds (amino derivatives of sugar) include, for example, glucosamine, galactosamine, 2-deoxy-2-methylaminoglucose and the like. The oligosaccharide compounds include non-reducing oligosaccharides and reducing oligosaccharides. Specific examples thereof include sucrose, trehalose, gentianose, raffinose, lactose, cerbiose, maltose, and gentiobiose.

  The sugar compound is usually bonded through an oxygen atom bonded to the terminal 1-position carbon atom of the sugar skeleton.

The organic groups R ¹ to R ³ in the present invention may be an amine compound residue. In this amine compound, the carbon number is 50 or less, preferably 35 or less, more preferably 2 to 12. The ratio of nitrogen atoms is the ratio [N] / [C] of the number of nitrogen atoms [N] to the number of carbon atoms [C], and is 1/50 to 2/1, preferably 1/35 to 1.
This amine compound is usually bonded via its nitrogen atom.

  The amine compound includes amino acid compounds and peptide compounds in addition to primary to tertiary amines.

  Specific examples of amino acid compounds include glycine, alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, glutamic acid, asparagine, glutamine, lysine, ornithine, arginine, histidine, hydroxylysine, cysteine, cystine, methionine, phenylalanine, Tyrosine, tryptophan, proline, 4-hydroxyproline, tricolominic acid, ipotenic acid, kisscaric acid, canavanine, kainic acid, domoic acid, 1-aminocyclopropanecarboxylic acid, 2- (methylenecyclopropyl) glycine, hypoglycine A, 3- Examples include cyanoalanine, abenaic acid, mugineic acid, mimosine, levodopa, β-hydroxy-γ-methylflutamic acid, 5-hydroxytryptophan, pantothenic acid, laminin, betacyanin Indicated. In addition, amine compounds having a sulfonic acid group such as taurine are also included.

  The amine compound (including an amino acid compound and a peptide compound) can contain a halogen atom. The amine compound may have a structure in which a phosphate group and an amino alcohol (choline, ethanolamine, serine, etc.) are condensed.

In the present invention, the organic groups R ¹ to R ³ may be nucleic acid compound residues. In this nucleic acid compound, the carbon number is 150 or less, preferably 100 or less, more preferably 4 to 50.
This nucleic acid compound is usually bound via its oxygen, nitrogen or carbon atom.

  Nucleic acid compounds include nucleosides, nucleotides, or polymer chains of nucleotide units. Examples of the nucleoside include those in which a base contained in cytosine, thymine, adenine, and guanine and a sugar of 2-deoxyribose or ribose are bonded. As the nucleotide, a nucleoside sugar bonded to a phosphate group can be shown. As thymine, uracil can be shown.

In the above general formula, X ^{1 to} X ⁴ each independently represents a methylene group or a carbonyl group. When X ^{1 to} X ⁴ are methylene groups, the compound of the present invention is glycerol and organic groups R ^{1 to} R ³ having an ether bond. When X ^{1 to} X ⁴ are carbonyl groups, glycerol and the organic group R ^{1 to} R ³ are ester-bonded. Many natural phospholipids contain an ester bond, but it is known that some contain an ether bond. It is well known to those skilled in the art that its physicochemical properties are very similar except for the presence or absence of resistance to degrading enzymes, and the synthesis of ether type lipids is also known. Also in the compound of the present invention, an ester bond type and an ether bond type can be synthesized, respectively, and can be used similarly.

In the above general formula, Y ¹ and Y ² each independently represent a hydrogen atom, phosphoric acid, phosphocholine, phosphoserine, phosphoethanolamine or phosphoglycerol. These groups are known as groups constituting the hydrophilic portion of natural phospholipids, and these groups can be included in the compounds of the present invention as well. More preferably Y ¹ and Y ² are a hydrogen atom, phosphoric acid, phosphocholine or phosphoglycerol, particularly preferred are phosphoric acid or phosphocholine.

  In the present invention, a particularly suitable compound is not particularly limited, but examples thereof include compounds 11 to 13, 21 and 35 having the following formulas shown in the examples.

The compound 11 is one of the compounds represented by the general formula (1), and includes two triple bonds in R ¹ and 9 fluorine atoms (4 perfluorocarbons) in R ² and R ^3. Alkyl group). In compound 11, X ^{1 to} X ⁴ are methylene groups, and Y ¹ and Y ² are all hydrogen atoms.

The compound 12 is one of the compounds represented by the general formula (1) like the compound 11, and includes two triple bonds in R ¹ and 9 fluorine atoms (carbon) in R ² and R ³ respectively. Several perfluoroalkyl groups). In compound 12, X ^{1 to} X ⁴ are methylene groups, and Y ¹ and Y ² are all phosphocholines.

The compound 13 is one of the compounds represented by the general formula (1), and each of R ² and R ³ contains 9 fluorine atoms (a perfluoroalkyl group having 4 carbon atoms). In compound 13, X ^{1 to} X ⁴ are methylene groups, and Y ¹ and Y ² are all phosphocholines.

The compound 21 is one of the compounds represented by the general formula (2), and includes 16 fluorine atoms (a perfluoroalkylene group having 8 carbon atoms) in R ¹ . In compound 21, X ^{1 to} X ⁴ are methylene groups, and Y ¹ and Y ² are all hydrogen atoms.

The compound 35 is one of the compounds represented by the general formula (3), and includes two triple bonds in R ¹ , 17 fluorine atoms (perfluoroalkyl group having 8 carbon atoms) in R ² , and It contains one triple bond and R ³ contains one triple bond. In the compound 35, X ¹ , X ² and X ⁴ are methylene groups, X ³ is a carbonyl group, and Y ¹ and Y ² are all hydrogen atoms.

  A person skilled in the art can appropriately obtain the compound of the present invention as necessary based on the description in the present specification and the common general technical knowledge in the field of chemical synthesis. It will be understood that this is not a limitation.

  The pseudo-cyclic lipid compound of the present invention that can be represented by the general formulas (1) to (3) can be used for various purposes by utilizing its surface activity alone. In particular, it can contribute to the improvement of membrane characteristics as a constituent of the reconstituted lipid membrane. The characteristics of the compound of the present invention vary depending on the carbon chain length, the presence or absence of fluorine or a triple bond, the nature of each functional group, etc., and are difficult to define strictly. Those having a chain length are suitable as a surfactant, while those having a chain length of 12 or more carbon atoms may be suitable as a membrane component.

  In addition, the present invention can create a molecular assembly composed of the quasicyclic lipid compound of the present invention, or a molecular assembly composed of one or more quasicyclic lipid compounds and one or more different lipids. Moreover, proteins can also be included in these molecular assemblies.

  In this specification, the term “molecular assembly” refers to a mesoscale structure in which a large number of amphiphilic molecules are assembled by non-covalent interactions, such as a spherical shell or disk-shaped bicell or ribbon. There is a possibility that a molecular assembly such as a shape is formed. Micelles, liposomes, and the like are also included in the molecular assembly referred to in the present invention.

  The molecular assembly composed of the quasi-cyclic lipid compound of the present invention improves the membrane stability by quasi-cyclizing the compound, and further improves the membrane stability by fluorination. In addition, in natural lipids, the introduction of unsaturated bonds into pseudocyclic lipid compounds can improve the handling surface, just as the presence of unsaturated bonds lowers the melting point and facilitates handling. .

  The quasi-cyclic lipid compound has a much higher gel-liquid crystal phase transition temperature compared to the bilayer membrane from the corresponding double-chain lipid. This indicates that the pseudocyclic lipid compound forms a very stable lipid membrane. In addition, a certain amount of fluorination is expected to improve the stable gel-liquid crystal phase transition temperature.

  Compared to the case where the double bond is once synthesized via the triple bond, the triple bond is easier to synthesize and is less susceptible to the oxidation reaction that causes problems with the double bond (such as oleic acid). Have advantages. In addition, since the triple bond shows the rigidity of the structure due to the linear structure of four carbons, it is rarely used for lipids that require moderate flexibility such as vesicles (J. Fluorine Chem., 128, 133-138 (2007)).

  The pseudo-cyclic lipid compound of the present invention can be a component of a lipid membrane (molecular assembly) alone. Moreover, as one embodiment, when the pseudo-cyclic lipid compound of the present invention is used as a component of a lipid membrane, it can be prepared as a molecular assembly such as a liposome together with other natural or synthetic lipids.

  In the present specification, “lipid” sometimes represents a complex lipid in which phosphoric acid, sugar, amino alcohol, and the like are added to both elements of fatty acid and alcohol.

  The complex lipid represents a lipid having 3 to 50 carbon atoms, preferably 3 to 36 carbon atoms, and may contain oxygen, nitrogen, phosphorus, and sulfur. For example, glycerophospholipids such as lecithin, kephalin, phosphatidylserine, phosphatidylinositol, plasmalogen, dimyristoylphosphatidylcholine, alkyl ether glycerophospholipids, sphingophospholipids such as sphingosine and sphingomyelin, glyceroglycolipids as digalactosyl diglyceride Examples of glycosphingolipids such as 6-sulfoquinovosyl diglyceride include galactosylceramide, lactosylceramide, hematoside, and phytosphingosine.

  Moreover, the compound of the present invention can constitute a lipid membrane together with other lipids such as cholesterol, natural or synthetic membrane proteins, etc. in addition to the above-described phospholipids.

  The suitable ratio of the compound of the present invention to other lipids in the lipid membrane varies depending on the type of the compound of the present invention used and other lipids, but generally ranges from 1:99 to 100: 0, preferably A range of 10:90 to 90:10 (molar ratio) is preferred. Within this range, the properties such as the lipid bilayer in the liposome can be maintained, and effects such as the construction of lipid spheres containing an aqueous or lipid-soluble compound group can be obtained.

  The membrane protein may be any protein that is naturally attached to a biological membrane, and may be a protein that binds to and partially penetrates a lipid membrane, or a transmembrane protein. Examples include receptors such as GPCR (G protein-coupled receptor), enzymes such as hydrolases, photosynthetic membrane proteins and light-driven ion transport proteins, and the bacteriorhodopsin described in the following examples is also a membrane protein. One type.

  The membrane stabilization effect of the compound of the present invention is, for example, that a molecular assembly containing the compound of the present invention, for example, a liposome is prepared containing bacteriorhodopsin, and its CD spectrum is measured to stabilize the trimer structure of bacteriorhodopsin. It can be done by checking whether or not it is held.

Methods for Synthesizing the Compounds of the Invention The compounds of the invention can be readily synthesized by those skilled in the art based on the description herein and knowledge of organic chemical synthesis known in the art.

Specifically, the compound of the present invention can be synthesized through the following steps.
Preparation of chiral compounds Chiral glycerol derivatives are for example (S)-(+)-2,2-dimethyl-1,3-dioxolane-4-methanol or (R)-(-)-2,2-dimethyl- Start with 1,3-dioxolane-4-methanol or (R)-(+)-3-benzyloxy-1,2-propanediol or (S)-(-)-3-benzyloxy-1,2-propanediol It can be synthesized by using it as a substance. Alternatively, it can be obtained by optical resolution of a racemic mixture or resolution using an enzyme.

  In one embodiment of the present invention, a membrane protein that is a biological material is used. In this case, it is considered preferable that the compound is chiral. When it is not necessary to use a chiral compound, a mixture of two chiral reagents is commercially available and can be synthesized in the same manner.

Introduction of an aliphatic group Introduction of an aliphatic group is performed by using an alcohol, an alkyl halide or a carboxylic acid having a carbon chain having a desired chain length, such as 9-decen-1-ol, 1-bromotetradecane, 14-tetradecynoic acid, etc. It can be performed by reacting with. Cyclic aliphatic groups can be introduced by reacting with cyclic aliphatic alcohols, halogenated compounds, carboxylic acids, and the like, such as cyclopentenol, bromocyclooctane, cyclohexyl carboxylic acid, and the like.

Introduction of an aromatic group Introduction of an aromatic group can be performed by reacting with an aromatic alcohol having a desired carbon chain, a halogenated product, a carboxylic acid, etc., such as a bisphenol compound, bromophenol, benzoic acid, or the like. .

Introduction of a heterocyclic group Introduction of a heterocyclic group can be achieved by, for example, alcohols and carboxylic acids having a heterocyclic ring having a target carbon chain, such as 5-hydroxyindole and 6-chloro-2,3-dihydro-3-oxo- This can be performed by reacting with 4H-1,4-benzoxazine-4-propionic acid or the like.

Introduction of sugar Sugar can be introduced through an oxygen atom bonded to the terminal carbon atom of the sugar skeleton such as glucose, glucosamine or sucrose having a target carbon chain.

Introduction of an amine compound Introduction of an amine compound is carried out by introducing an amino acid or peptide compound having a target carbon chain, such as serine, aspartame, collagen, etc., with an ester bond at the C-terminus and N, N'-disuccinimidyl carbonate at the N-terminus It can be carried out by ester bonding through the like.

Nucleic acid introduction Nucleic acid introduction should be performed by N-N'-disuccinimidyl carbonate, etc., or phosphate ester bonding of the amino group of a nucleotide or deoxynucleotide with a nucleic acid, sugar and phosphate group. I can do it.

Protection and deprotection of active groups As is well known in the art, other reactive groups are pre-protected to ensure that only the desired reaction occurs at each step. For example, p-methoxybenzyl chloride, trityl chloride, or the like is used, and a hydroxyl group that is not intended to be reacted is previously protected and deprotected in a later step.

Introduction of fluorine Introduction of a fluorine group is carried out, for example, by reacting with perfluorobutyl iodide in order to introduce a perfluoroalkyl group. When a fluorine group is introduced in the middle of the molecule, another group such as an alkyl group is further bonded, or hexadecafluoro-1,8-diiodooctane and By reacting, introduction of fluorine and dimerization can be performed simultaneously. The length of the perfluoro group to be introduced can be appropriately changed by selecting a reagent to be reacted, and such a reagent is commercially available or can be obtained by synthesis. A person skilled in the art can easily determine the reaction conditions for introducing fluorine. It will be understood by those skilled in the art that the film stability of the resulting compound varies depending on the length of the perfluoro group.

Introduction of a triple bond Introduction of a triple bond can be achieved, for example, by reacting a commercially available reagent having a triple bond such as 7-hexadecin-1-ol or a compound obtained by an acetylene introduction reaction. One or two triple bonds may be included in any one or more of R ^{1 to} R ³ , respectively.

Although the manufacturing method of the compound of this invention is not specifically limited, Pseudo-cyclization can be carried out by couple | bonding the same or different glycerol derivative. This can be performed, for example, by a diacetylene coupling reaction, an esterification reaction, an etherification reaction, or the like.

Phosphorylation / choline phosphorylation represents a phosphate ester, and an ester bond can be easily synthesized from dehydration condensation of phosphoric acid and an alcohol compound. Further, choline can be easily synthesized by substituting one hydroxyl group of phosphoric acid with a choline group. Similarly, one hydroxyl group of phosphoric acid can be substituted with serine group, ethanolamine group, or glycerin.

Purification The compound of the present invention can be easily purified by, for example, silica gel column chromatography. A person skilled in the art can appropriately select the type of gel to be used, solvent, elution conditions, and the like.

Identification of Compound A compound can be identified by, for example, NMR. In the case of a compound containing fluorine, NMR can be performed by ¹⁹ F-NMR in addition to ¹ H-NMR.

  Hereinafter, the present invention will be described with reference to examples, but the present invention is not intended to be limited to these examples.

[Example 1]

Experiment 1
Under a nitrogen atmosphere and under water cooling, potassium hydroxide (10 eq), p-methoxy was added to a solution of (S)-(+)-2,2-dimethyl-1,3-dioxolane-4-methanol (1 eq) in dimethyl sulfoxide. Benzyl chloride (1.1-1.2 equivalents) was added and stirred at room temperature for 18 hours. The reaction solution was poured into ice water and extracted with diethyl ether. The diethyl ether extract was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20% ethyl acetate / hexane). To a methanol solution of the compound obtained, p-toluenesulfonic acid hydrate (catalytic amount) was added. In addition, the mixture was stirred at room temperature for 1-2 days. The solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (50-100% ethyl acetate / hexane) to obtain Compound 1 (74-94%, 2 steps).

Experiment 2
Under a nitrogen atmosphere, trityl chloride (1.1 eq) was added to an anhydrous dichloromethane solution of compound 1 (1 eq), triethylamine (2 eq) was added under ice cooling, and the mixture was stirred at room temperature for 18 hours. The reaction solution was poured into ice-cooled 10% aqueous hydrochloric acid and extracted with dichloromethane. The dichloromethane extract was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20-50% ethyl acetate / hexane) to obtain Compound 2 (86%).

Experiment 3
Under a nitrogen atmosphere, methanesulfonyl chloride (1.5 equivalents) is added to an anhydrous dichloromethane solution of 9-decen-1-ol (1 equivalent), and triethylamine (2 equivalents) is added under ice cooling, followed by stirring at room temperature for 3 hours. did. The reaction solution was poured into ice-cooled 10% aqueous hydrochloric acid and extracted with dichloromethane. The dichloromethane extract was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20% ethyl acetate / hexane) to obtain Compound 3 (95% or more).

Experiment 4
Under a nitrogen atmosphere, sodium hydride (40% mineral oil added, 1.5 equivalents) was added to an anhydrous N, N-dimethylformamide solution of compound 2 (1 equivalent) under ice cooling, and after 10 minutes, 30 minutes at room temperature. Stir for minutes. Under ice cooling, an anhydrous DMF solution of Compound 3 (1.5 equivalents) was added dropwise. Tetrabutylammonium iodide (catalytic amount) was added, and the mixture was stirred at room temperature for 18 hours. After adding ice water, the mixture was extracted with diethyl ether, washed with saturated brine, and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (10% ethyl acetate / hexane) to obtain Compound 4 (66-76%).

Experiment 5
Under a nitrogen atmosphere, p-toluenesulfonic acid hydrate (catalytic amount) was added to a chloroform-methanol solution of compound 4 (1 equivalent), and the mixture was stirred at room temperature for 18 hours. The solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20-30% ethyl acetate / hexane) to give compound 5 (85-95%).

Experiment 6
Under a nitrogen atmosphere, perfluorobutyl iodide (1.2 eq), sodium bicarbonate (0.5 eq) and sodium hyposulfite (0.5 eq) are added to an acetonitrile-water solution of compound 5 (1 eq), and the mixture is brought to room temperature. And stirred for 4 hours. After dilution with water, extraction with dichloromethane was performed. The dichloromethane extract was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (30-50% ethyl acetate / hexane) to obtain Compound 6 (83-93%).

Experiment 7
To a solution of zinc (2 equivalents) in tetrahydrofuran, add nickel chloride hydrate (0.1 equivalent) and a few drops of water, stir at room temperature for 10 minutes, then add compound 6 (1 equivalent) in tetrahydrofuran and add at room temperature. Stir for 3 hours. Saturated sodium bicarbonate was added, filtered, and extracted with dichloromethane. The dichloromethane extract was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (50% ethyl acetate / hexane) to obtain Compound 7 (83 to 93%).

Experiment 8
Under a nitrogen atmosphere, methanesulfonyl chloride (1.5 equivalents) was added to an anhydrous dichloromethane solution of 13-tetradecin-1-ol (1 equivalent), and triethylamine (2 equivalents) was added under ice cooling, followed by stirring at room temperature for 3 hours. did. The reaction solution was poured into ice-cooled 10% aqueous hydrochloric acid and extracted with dichloromethane. The dichloromethane extract was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (30% ethyl acetate / hexane) to obtain Compound 8 (95% or more).

Experiment 9
Under a nitrogen atmosphere, sodium hydride (40% mineral oil added) was added to an anhydrous N, N-dimethylformamide solution of Compound 7 (1 equivalent) under ice cooling, and the mixture was stirred for 10 minutes and then at room temperature for 30 minutes. Under ice cooling, an anhydrous DMF solution of compound 8 (1.5 equivalents) was added dropwise. Tetrabutylammonium iodide (catalytic amount) was added, and the mixture was stirred at room temperature for 18 hours. After adding ice water, the mixture was extracted with diethyl ether, washed with saturated brine, and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20% ethyl acetate / hexane) to obtain Compound 9 (36-%).

Experiment 10
2,3-Dichloro-5,6-dicyano-p-benzoquinone (2 equivalents) was added to a mixed solution of compound 9 (1 equivalent) in dichloromethane-phosphate buffer (pH 7) under ice cooling, and the mixture was stirred for 2 hours. . Saturated aqueous sodium hydrogen carbonate was added, extracted with dichloromethane, washed with saturated brine, and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20% ethyl acetate / hexane) to obtain Compound 10 (96%).

Experiment 11
Copper chloride was added to a piperidine solution of compound 10 (1 equivalent), and the mixture was stirred at 140 degrees for 30 minutes. After adding ice water, the mixture was extracted with chloroform, washed with saturated brine, and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (40% ethyl acetate / hexane) to obtain Compound 11 (94%). From the following NMR measurement values, it was confirmed that the product was the compound 11.
¹ H-NMR (CDCl ₃ , TMS): 3.38-3.77 (m, 18H), 2.34 (brs, 2H), 2.24 (t, J = 6.98 Hz, 4H); 05 (m, 4H), 1.46-1.68 (m, 16H), 1.19-1.45 (m, 56H).
¹⁹ F-NMR (CDCl ₃ , CFCl ₃ ): −81.63 (m, 6F), −115.12 (m, 4F), 125.01 (m, 4F), −126.58 (m, 4F) .

Experiment 12
Triethylamine (2.5 equivalents) and 2-bromoethylphosphorodichloridate (1.5 equivalents) were added to a benzene solution of compound 11 (1 equivalent) and stirred at room temperature for 18 hours. The solvent was distilled off under reduced pressure, water was added, and the mixture was stirred at room temperature for 4 hours. After extraction with chloroform, the solvent was distilled off under reduced pressure, and a 2-propanol-acetonitrile-chloroform mixture and a 30% trimethylamine aqueous solution were added to the residue, followed by stirring at 60 ° C. for 18 hours. The solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (chloroform / methanol / water) and ODS column (chloroform / methanol) to obtain Compound 12 (85%). From the following NMR measurement values, it was confirmed that the product was the compound 12.
¹ H-NMR (CDCl ₃ , TMS): 4.27 (brs, 4H), 3.90 (m, 4H), 3.55 to 3.70 (m, 12H), 3.43 to 3.53 ( m, 6H), 3.32 (s, 18H), 2.24 (t, J = 6.89 Hz, 4H), 2.08 (m, 4H), 1.46-1.70 (m, 16H) 1.23-1.46 (m, 56H).
¹⁹ F-NMR (CDCl ₃ , CFCl ₃ ): −81.34 (m, 6F), −115.53 (m, 4F), 124.43 (brs, 4F), −126.11 (m, 4F) .

Experiment 13
10% Pd / C (catalytic amount) was added to a methanol solution of compound 12 (1 equivalent). The mixture was stirred at room temperature for 18 hours under a hydrogen atmosphere and then filtered. The solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (chloroform / methanol / water) and ODS column (chloroform / methanol) ODS column (chloroform / methanol) to obtain compound 13 (54%). It was. From the following NMR measurement values, it was confirmed that the product was the compound 13.
¹ H-NMR (CDCl ₃ , TMS): 4.28 (brs, 4H), 3.89 (t, J = 5.54 Hz, 4H), 3.51 to 3.67 (m, 12H), 3. 39-3.50 (m, 6H), 3.20 (s, 18H), 2.10 (m, 4H), 1.50-1.67 (m, 12H), 1.20-1.47 ( m, 72H).
¹⁹ F-NMR (CDCl ₃ , CFCl ₃ ): −81.13 (m, 6F), −114.25 (m, 4F), 124.11 (m, 4F), −125.84 (m, 4F) .

[Example 2]

Experiment 14
Under a nitrogen atmosphere and water cooling, potassium hydroxide (10 eq), p-methoxy was added to a solution of (R)-(−)-2,2-dimethyl-1,3-dioxolane-4-methanol (1 eq) in dimethyl sulfoxide. Benzyl chloride (1.1-1.2 equivalents) was added and stirred at room temperature overnight. The reaction solution was poured into ice water and extracted with diethyl ether. The diethyl ether extract was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20-30% ethyl acetate / hexane) to a methanol solution of the compound obtained, and p-toluenesulfonic acid hydrate (catalytic amount). ) And stirred at room temperature to 90 degrees for 3 hours to 2 days. The solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (50-100% ethyl acetate / hexane) to obtain Compound 14 (58-78%, 2 steps).

Experiment 15
Under a nitrogen atmosphere, trityl chloride (1.1 eq) was added to an anhydrous dichloromethane solution of compound 14 (1 eq), triethylamine (2 eq) was added under ice cooling, and the mixture was stirred overnight at room temperature. The reaction solution was poured into ice-cooled 10% aqueous hydrochloric acid and extracted with dichloromethane. The dichloromethane extract was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (30% ethyl acetate / hexane) to obtain Compound 15 (74-99%).

Experiment 16
Under a nitrogen atmosphere, sodium hydride (40% mineral oil added, 1.5 equivalents) was added to a solution of compound 15 (1 equivalent) in anhydrous N, N-dimethylformamide under ice-cooling, 10 minutes later, 30 minutes at room temperature. Stir for minutes. Under ice cooling, an anhydrous N, N-dimethylformamide solution of compound 3 (1.5 equivalents) was added dropwise. Tetrabutylammonium iodide (catalytic amount) was added, and the mixture was stirred overnight at room temperature. After adding ice water, the mixture was extracted with diethyl ether, washed with saturated brine, and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (10-30% ethyl acetate / hexane) to obtain Compound 16 (93%).

Experiment 17
Under a nitrogen atmosphere, 2,3-dichloro-5,6-dicyano-p-benzoquinone (2 equivalents) was added to a mixed solution of compound 16 (1 equivalent) in dichloromethane-phosphate buffer (pH 7) under ice cooling. The mixture was stirred from ice-cooled to room temperature for 2 hours to overnight. After adding ice water, the mixture was extracted with dichloromethane, washed with saturated brine, and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20% ethyl acetate / hexane) to obtain Compound 17 (78-88%).

Experiment 18
Under a nitrogen atmosphere, sodium hydride (40% mineral oil added, 1.5-5.0 equivalents) in anhydrous N, N-dimethylformamide solution under ice-cooling, compound 17 (1 equivalent) in anhydrous N, N-dimethyl A formamide solution was added, and the mixture was stirred at ice-cooled to room temperature for 10 to 30 minutes. Then, an ice-cooled solution of 1-bromotetradecane (1.5 equivalents) in anhydrous N, N-dimethylformamide was added and stirred at room temperature for 18 hours. did. The reaction solution was poured into ice-cooled 10% aqueous hydrochloric acid and extracted with diethyl ether. The diethyl ether extract was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (10% ethyl acetate / hexane) to obtain Compound 18 (48-68%).

Experiment 19
P-Toluenesulfonic acid hydrate (catalytic amount) was added to a chloroform-methanol solution of compound 4 (1 equivalent), and the mixture was stirred at room temperature for 18 hours. The solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20% ethyl acetate / hexane) to obtain Compound 19 (86%).

Experiment 20
Add hexadecafluoro-1,8-diiodooctane (1 equivalent), sodium hydrogen carbonate (0.5 equivalent), and sodium hyposulfite (0.5 equivalent) to an acetonitrile-water solution of compound 19 (2.2 equivalents). And stirred at 50 ° C. for 4 hours. After dilution with water, extraction with dichloromethane was performed. The dichloromethane extract was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (ethyl acetate / hexane / chloroform) to obtain Compound 20 (58%). From the following NMR measurement values, it was confirmed that the product was the compound 20.
¹ H-NMR (CDCl ₃ , TMS): 4.33 (m, 2H), 3.40-3.76 (m, 18H), 2.67-3.02 (m, 4H), 2.37 ( brs, 2H), 1.70-1.89 (m, 4H) 1.48-1.66 (m, 8H), 1.17-1.48 (m, 64H) 0.88 (t, J = 6.68 Hz, 6H).
¹⁹ F-NMR (CDCl ₃ , CFCl ₃ ): −112.23 (dm, J = 270.42 Hz, 2F), −115.10 (dm, J = 270.42 Hz, 2F), 122.06 ( br s, 4F), -122.36 (br s, 4F), -124.13 (br s, 4F).

Experiment 21
Sodium hydrogen carbonate, 10% Pd / C (catalytic amount) was added to a methanol-chloroform solution of compound 20 (1 equivalent). Under a hydrogen atmosphere, the mixture was stirred at room temperature for 24 hours and then filtered. The solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (ethyl acetate / hexane / chloroform) to obtain Compound 21 (65%). From the following NMR measurement values, it was confirmed that the product was the compound 21.
¹ H-NMR (CDCl ₃ , TMS): 3.40-3.77 (m, 18H), 2.31 (brs, 2H), 2.43 (m, 4H), 1.50-1.65 ( m, 12H), 1.27-1.48 (m, 68H), 0.88 (t, J = 6.80 Hz, 6H).
¹⁹ F-NMR (CDCl ₃ , CFCl ₃ ): −114.91 (quin, J = 14.29 Hz, 4F), −122.26 (brs, 4F), −122.44 (brs, 4F), −124 .10 (brs, 4F).

[Example 3]

Experiment 22
Under a nitrogen atmosphere, methanesulfonyl chloride (1.5 equivalents) was added to an anhydrous dichloromethane solution of 15-hexadecin-1-ol (1 equivalent), and triethylamine (2 equivalents) was added under ice cooling, followed by stirring at room temperature for 3 hours. did. The reaction solution was poured into ice-cooled 10% aqueous hydrochloric acid and extracted with dichloromethane. The dichloromethane extract was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20% ethyl acetate / hexane) to obtain Compound 22 (94% or more).

Experiment 23
Under a nitrogen atmosphere, sodium hydride (mineral) was added to an anhydrous N, N-dimethylformamide solution of (R)-(−)-2,2-dimethyl-1,3-dioxolane-4-methanol (1 equivalent) under ice-cooling. (40% oil, 1.5 equivalents) was added, and after 10 minutes, the mixture was stirred at room temperature for 30 minutes. Under ice cooling, an anhydrous N, N-dimethylformamide solution of compound 22 (1.5 equivalents) was added dropwise. Tetrabutylammonium iodide (catalytic amount) was added, and the mixture was stirred at room temperature for 8 to 24 hours. After adding ice water, the mixture was extracted with diethyl ether, washed with saturated brine, and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (10-20% ethyl acetate / hexane) to obtain Compound 23 (83%).

Experiment 24
P-Toluenesulfonic acid hydrate (catalytic amount) was added to a methanol solution of compound 23, and the mixture was stirred at room temperature to 90 degrees for 3 hours to 3 days. The solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (50-100% ethyl acetate / hexane) to obtain Compound 24 (73-92%).

Experiment 25
Under a nitrogen atmosphere, trityl chloride (1.1 eq) was added to an anhydrous dichloromethane solution of compound 24 (1 eq), triethylamine (2 eq) was added under ice cooling, and the mixture was stirred overnight at room temperature. The reaction solution was poured into ice-cooled 10% aqueous hydrochloric acid and extracted with dichloromethane. The dichloromethane extract was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (10-20% ethyl acetate / hexane) to obtain compound 25 (90-94%).

Experiment 26
Compound 26 (11,11,12,12,13,13,14,14,15,15,16,16,17,17,18,18,18-heptadecafluoro-9-octadecanoic acid (heptadecafluorooctadec-9- ynoic acid)) was synthesized with reference to the literature (K. Takai, T. Takagi, T. Baba, T. Kanamori, J. Fluorine Chem., 124, 1959 (2004)).

Experiment 27
To a solution of compound 25 (1 eq) in anhydrous dichloromethane, compound 26 (1.5-2 eq), N, N′-dicyclohexylcarbodiimide (1.2 eq), 4-dimethylaminopyridine (1.2 eq) were added, Stir at room temperature overnight. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (10% ethyl acetate / hexane) to obtain Compound 27 (crude).

Experiment 28
P-Toluenesulfonic acid hydrate (catalytic amount) was added to a methanol-chloroform solution of Compound 27, and the mixture was stirred overnight at room temperature. The solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20-30% ethyl acetate / hexane) to obtain compound 28 (48-58%, 2 steps).

Experiment 29
Under a nitrogen atmosphere, sodium hydride (40% mineral oil added, 1.5 equivalents) was added to an anhydrous N, N-dimethylformamide solution of compound 15 (1 equivalent) used in Example 2 under ice cooling, and 10 minutes. Thereafter, the mixture was stirred at room temperature for 30 minutes. Under ice cooling, an anhydrous N, N-dimethylformamide solution of compound 22 (1.5 equivalents) was added dropwise. Tetrabutylammonium iodide (catalytic amount) was added, and the mixture was stirred at room temperature for several hours to 3 days. After adding ice water, the mixture was extracted with diethyl ether, washed with saturated brine, and dried over anhydrous magnesium sulfate. . After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (10-30% ethyl acetate / hexane) to obtain compound 29 (70-80%).

Experiment 30
P-Toluenesulfonic acid hydrate (catalytic amount) was added to a methanol-chloroform solution of Compound 29, and the mixture was stirred at room temperature for 1 to 3 days. The solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (30-40% ethyl acetate / hexane) to obtain compound 30 (46-56%).

Experiment 31
Under a nitrogen atmosphere, methanesulfonyl chloride (1.5 equivalents) was added to an anhydrous dichloromethane solution of 7-hexadecin-1-ol (1 equivalent), and triethylamine (2 equivalents) was added under ice cooling, followed by stirring at room temperature for 3 hours. did. The reaction solution was poured into ice-cooled 10% aqueous hydrochloric acid and extracted with dichloromethane. The dichloromethane extract was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20-30% ethyl acetate / hexane) to obtain Compound 31 (88%).

Experiment 32
Under a nitrogen atmosphere, sodium hydride (40% mineral oil added, 1.5 equivalents) was added to an anhydrous N, N-dimethylformamide solution of compound 30 (1 equivalent) under ice-cooling, 10 minutes later, 30 minutes at room temperature. Stir for minutes. Under ice cooling, an anhydrous N, N-dimethylformamide solution of compound 31 (1.5 equivalents) was added dropwise. Tetrabutylammonium iodide (catalytic amount) was added, and the mixture was stirred overnight at room temperature. After adding ice water, the mixture was extracted with diethyl ether, washed with saturated brine, and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (10-30% ethyl acetate / hexane) to obtain Compound 32 (55-65%).

Experiment 33
Under a nitrogen atmosphere, 2,3-dichloro-5,6-dicyano-p-benzoquinone (2 equivalents) was added to a mixture of Compound 32 (1 equivalent) in dichloromethane-phosphate buffer (pH 7) under ice cooling. The mixture was stirred on ice for 4 hours. After adding ice water, the mixture was extracted with dichloromethane, washed with saturated brine, and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20-30% ethyl acetate / hexane) to obtain Compound 33 (93%).

Experiment 34
In a nitrogen atmosphere, iodine (2 equivalents) was added to a benzene solution of morpholine (6 equivalents) and stirred at room temperature for 30 minutes. A benzene solution of compound 33 (1 equivalent) was added, and the mixture was stirred at 45 degrees for a whole day and night. An aqueous sodium thiosulfate solution was added, and the mixture was extracted with chloroform, washed with saturated brine, and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20-30% ethyl acetate / hexane) to obtain Compound 34 (71%).

Experiment 35
Under a nitrogen atmosphere, copper (I) iodide (catalytic amount) was added to a pyrrolidine solution of compound 28 and compound 34, and the mixture was stirred at room temperature for 30 minutes. After adding an aqueous ammonium chloride solution, the mixture was extracted with ether, washed with saturated brine, and dried over anhydrous magnesium sulfate. After filtration, the solvent was distilled off under reduced pressure, and the residue was purified by passing through silica gel column chromatography (20-30% ethyl acetate / hexane) to obtain Compound 35 (63%). From the following NMR measurement values, it was confirmed that the product was the compound 35.
¹ H-NMR (CDCl ₃ , TMS): 4.16 (td, J = 11.52, 4.34 Hz, 1H), 4.13 (td, J = 11.52, 6.20 Hz, 1H), 4 .00 (m, 1H), 3.38-3.77 (m, 13H), 2.56 (d, J = 4.28 Hz, 1H), 2.30-2.43 (m, 4H), 2 19-2.28 (m, 5H), 2.09-2.18 (m, 4H), 1.17-1.70 (m, 78H), 0.88 (t, J = 6.86 Hz, 3H).
¹⁹ F-NMR (CDCl ₃ , CFCl ₃ ): −81.25 (tm, J = 9.63 Hz, 3F), −96.68 (m, 2F), −121.59 (m, 2F), −122 .21 to -122.63 (m, 4F), -123.04 (m, 2F), -123.22 (m, 2F), -126.60 (m, 2F).

[Example 4]
Using the compound 13 which is the pseudo-cyclic lipid compound obtained in Example 1 and the compound 36 containing carbon instead of fluorine in the compound 13, a complex containing these compounds and a membrane protein was created, and membrane stabilization evaluation was performed. Went. The structures of compounds 13 and 36 are shown below. Compound 36 was synthesized by the synthesis method described in Langmuir 7 (11) 2415 (1991); Langmuir 8 (2) 637 (1992); Bull. Chem. Soc. Jpn., 70, 2545 (1997), and the like. The synthesis was conducted with reference to the synthesis method up to compound 13 (carbon chain lengths of 32 and 16 in the literature).

  DSC measurement (Seiko Instruments EXSTAR DSC6100) was performed on compounds 13 and 36, and compared with 1,2-ditetradecyl-glycero-3-phosphocholine (DTPC) corresponding to the monomer when compound 36 was dimerized. did. These transition temperatures are shown in Table 1 below. As can be seen from the results in Table 1, compound 36 has a transition temperature that is about twice as high as that of DTPC, but compound 13 of the present invention has a transition temperature that is brought to a physiological temperature by the introduction of a perfluoro group. It was suggested that a film having moderate flexibility can be constructed.

I. Preparation of lipid suspension Bring the cryopreserved lipid compound (compound 13 or 36) back to room temperature.
2. Weigh lipid compound on aluminum foil (7.5 μmol).
3. Dissolve the compound in chloroform solvent and transfer to eggplant flask.
4). Under a nitrogen atmosphere, chloroform is distilled off (water bath 40 ° C.), and lipid is attached to the inner wall of the flask in a film form.
5. Dry the lipid film under reduced pressure (about 3 hours).
6). Add several glass beads and 10 ml of 100 mM phosphate buffer and seal.
7. Shake for about 20 minutes in a thermostatic chamber set at 25 ° C.
8). Sonication is performed with a bath sonicator (30 to 60 ° C., 1 to several hours) to obtain a 0.75 mM lipid suspension (liposome).

II. Preparation of solubilized bacteriorhodopsin (bR) 1. 40 μM natural purple membrane (obtained by cultivating highly halophilic bacterium H. salinarum and purifying according to previous reports) and 20 mM TritonX-100 solution ( Both are mixed with 100 mM phosphate buffer) and shake at 25 ° C. for 15 hours.
2. After centrifugation (4 ° C., 15,000 to 100,000 g, 1 hour), the collected supernatant is designated as solubilized bR.

III. Reconstitution of bR into liposomes 1. Mix 20 μM solubilized bR and 0.75 mM lipid suspension and shake at 25 ° C. for 5 hours.
2. Add 0.6 g of Bio-Beads (manufactured by Bio-rad) and stir at 25-30 ° C. for 5 hours.
3. Add 0.6 g of Bio-Beads and stir with a stirrer at 28 ° C. for 15 hours.
4). The supernatant is collected using a syringe (inner diameter 1 mm) and used as a reconstituted sample.

IV. UV spectrum measurement of reconstituted sample From the result of UV measurement using the reconstituted sample of Compound 13, it was found that the yield of the reconstituted sample was 70-80%.

V. CD spectrum measurement
V-1. Sample preparation The reconstituted sample is applied to Vortex and then shaken at 25 ° C. for 20 minutes.
Aliquot 2 ml from the sample of 2.1 and perform sonication (30 ° C, 1 hour) with a sonicator.
The sample of 3.2 is put into a two-sided transmission cell, the UV spectrum is measured, and the measurement sample concentration is obtained (from the absorbance at this time, the measurement sample concentrations of the reconstituted samples of Compound 13 and Compound 36 are 7.0 and 3.0 μM, respectively) )

V-2. Measuring method Set 3 samples of V-1 to CD measuring instrument (JASCO J-820).
2. Nitrogen is allowed to flow into the CD measuring device, and the inside of the measurement chamber is replaced with nitrogen.
3. The temperature of the sample is controlled by a Peltier thermostat. In order to make the whole sample uniform at the measurement temperature, the sample is stirred with a stirrer. In addition, the measurement was started after waiting about 5 minutes after reaching the measurement temperature.
4). Measurement was started from the low temperature side (10 ° C), and the same sample was used up to the high temperature side (80 ° C).

V-3. Results The results are shown in FIGS. In the CD spectrum of the reconstructed sample of Compound 13, a negative peak was observed up to around 70 ° C. From this result, it can be seen that bR reconstituted in the lipid membrane (liposome) of compound 13 retains the trimer up to 70 ° C. On the other hand, in the CD spectrum of the reconstructed sample of Compound 36, the negative peak was confirmed from 40 ° C to 45 ° C.

  From this result, it was found that the thermal stability of the bR reconstructed film was improved by the introduction of the perfluoro group.

  According to the present invention, there is provided a quasi-cyclic compound in which two glycerols are formed in one molecule via one divalent organic group, two monovalent organic groups and four ester or ether bonds. Is done.

  The compounds according to the invention are surface-active and are used in the fields of metalworking, mining, surface finishing, cleaning and cleaning, cosmetics, medicine and food processing and cooking, especially in industry and household, especially emulsifiers, demulsifiers, cleaning agents. It is widely used for dispersants and hydrotropes.

Claims (4)

Pseudocyclic compounds represented by the following general formulas (1) to (3).

[Wherein, R ¹ represents a divalent chain aliphatic group having 12 to 36 carbon atoms, R ² and R ³ represent a monovalent chain aliphatic group having 7 to 36 carbon atoms, and R ¹ to R ^3] Any one or more of R ³ have one or more fluorine atoms, or one or more non-terminal and non-conjugated triple bonds, or both. R ² and R ³ may be the same or different. X ^{1 to} X ⁴ each independently represents a methylene group or a carbonyl group, and Y ¹ and Y ² each independently represent a hydrogen atom, phosphoric acid, phosphocholine, phosphoserine, phosphoethanolamine or phosphoglycerol. The steric structure of the glycerol moiety is represented by (R, R) (S, S) (R, S) (S, R) or a mixture thereof. ]   A molecular assembly composed of the quasicyclic compound according to claim 1, or a molecular assembly composed of the quasicyclic compound and one or more different lipids.   The molecular assembly according to claim 2, further comprising a protein.   A liposome comprising the quasi-cyclic compound according to claim 1 as a constituent component.

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