JPH0676459B2 - COMPOUND AND POLYMER HAVING NATURAL PHOSPHOLIPID-LIKE STRUCTURE, AND PROCESS FOR PRODUCING THE SAME - Google Patents

Description

TECHNICAL FIELD The present invention relates to a compound having a structure similar to a natural phospholipid (hereinafter, may be abbreviated as a compound having a natural phospholipid-like structure) and a polymer thereof. And a method for producing the compound having a structure similar to the natural phospholipid.

(Prior Art) Many kinds of phospholipids are contained in the living body. And
These phospholipids are closely related to various metabolic processes of the living body as constituents of cells such as cell membranes, are an energy source of brain tissue, and further transport and absorption of fat, blood coagulation, and food As is clear from the fact that it contributes to the perception of taste, it plays a very important role in maintaining the life of the living body.

Therefore, many attempts have been made to apply phospholipids to various artificial organs, cell fusion, enzyme immobilization, artificial cultivation, biosensors and the like.

The conventional phospholipids used in the above-mentioned attempts are
Mainly, natural phospholipids extracted from living bodies such as lecithin, phosphatidylethanolamine, phosphatidylserine and the like.

However, in order to use the phospholipid in the above-mentioned attempts, it should be a relatively high molecular weight substance, capable of forming a strong film, and economically obtainable. Was needed. In this respect, natural phospholipids are not preferable because it is extremely difficult to obtain a uniform and strong membrane because of their low molecular weight.

Then, as an alternative to the natural phospholipid, for example, there is a compound having a structure similar to the natural phospholipid represented by the following formula, which is disclosed in JP-A-63-222185 of the present applicant.

The compound having a structure similar to the natural phospholipid represented by the formula has the advantages that the compound itself can be economically synthesized and a stronger membrane can be obtained as compared with the natural phospholipid.

(Problems to be Solved by the Invention) However, the compound having a natural phospholipid-like structure disclosed in JP-A-63-222185 described above is a structure in which a phospholipid is directly amide-bonded to a methacryloyl group,
The polymerizable functional group was a methacrylamide type. For this reason, this compound can be polymerized by radiation polymerization, but there is a drawback that the polymerization does not proceed in the ordinary radical polymerization.

The present invention has been made in view of the above points, and an object of the present invention is to provide a compound having a natural phospholipid-like structure which can be easily polymerized not only by radiation polymerization but also by ordinary radical polymerization. To provide a polymer thereof and a method for producing a compound having a structure similar to the natural phospholipid.

(Means for Solving the Problems) In order to achieve this object, the inventor of this application has made various studies. As a result, this object has been achieved by using a compound having a natural phospholipid-like structure as a compound containing a methacrylic acid ester type functional group having good polymerization reactivity.

Therefore, the compound having a natural phospholipid-like structure, which is the first invention of this application, has the following general formula: (Each of R ₁ and R ₂ in the formula represents an alkyl group having 12 to 20 carbon atoms or an alkenyl group having 12 to 20 carbon atoms).

Further, the polymer having a natural phospholipid-like structure of the second invention according to this application has the following general formula: (In the formula, each of R ₁ and R ₂ represents an alkyl group having 12 to 20 carbon atoms or an alkenyl group having 12 to 20 carbon atoms. Further, n represents the degree of polymerization and is a value within the range of 100 to 500. ) Is represented by.

The method of producing a compound having a phospholipid-like structure of the first invention, which is the third invention of this application, comprises the steps of reacting methacryloyl chloride, propionic acid and sodium hydroxide in the presence of sodium hydroxide to synthesize methacryloxypropionic acid. And a step of synthesizing methacryloyloxypropionyl chloride by reacting this methacryloyloxypropionic acid with thionyl chloride in the presence of pyridine, and adding the following general formula to the methacryloyloxypropionyl chloride: (Wherein each of R ₁ and R ₂ represents an alkyl group having 12 to 20 carbon atoms or an alkenyl group having 12 to 20 carbon atoms) in the presence of pyridine. Characterize.

(Function) Since the compound having a phospholipid-like structure of the first invention of this application contains a methacrylic acid ester type polymerizable functional group having good polymerization reactivity, it is of course possible to polymerize by radiation. Radical-polymerize easily with a suitable initiator to give the polymer.

In addition, the polymer of the compound having a phospholipid-like structure of the second invention of this application has a high molecular weight, and thus exhibits a property that conventional natural phospholipids do not have.

According to the method for producing a compound having a phospholipid-like structure of the third invention of this application, the natural phospholipid-like compound of the first invention can be easily polymerized not only by radiation polymerization but also by ordinary radical polymerization. A compound having a structure can be easily produced.

(Example) Hereinafter, each invention which concerns on this application is demonstrated with the following example. The numerical conditions, chemicals used, and
It should be understood that the devices used and the like are merely examples for facilitating the understanding of the inventions, and therefore the inventions are not limited to these numerical conditions, chemicals used, devices used, and the like. .

Example 1 Example 1 is an example of a compound having a natural phospholipid-like structure of the first invention according to the production method of the third invention 2-
(Methacryloyloxy) ethyldipalmitoyl-DL-
α-phosphatidylethanolamide (hereinafter, referred to as Example 1
It may be abbreviated as the compound of. ), And further an example of producing a polymer of the compound of Example 1.

<Explanation of Compound of Example 1> Methacryloyl chloride and propionic acid are reacted in the presence of sodium hydroxide to synthesize methacryloxypropionic acid (hereinafter, sometimes abbreviated as compound (I)). The procedure described below was performed. The following equation (1) is the reaction equation.

First, in a three-necked flask equipped with a stirrer, a calcium chloride tube, and a dropping funnel, 20.0 g (0.22 mol) of 3-hydroxypropionic acid and 17.8 g (0.44 mol) of an aqueous solution of sodium hydroxide were dissolved (100 ml). ). Next, the flask was cooled to 0 ° C. in an ice bath, and the aqueous solution in the flask was stirred while being poured into the aqueous solution through a dropping funnel.
23.2 g (0.22 mol) of methacryloyl chloride is slowly added dropwise over 1 hour. The aqueous solution was further stirred at room temperature for 12 hours, and hydrochloric acid was added to adjust the pH of the aqueous solution to 2. Next, this solution is extracted with chloroform and dehydrated with anhydrous sodium sulfate. Then, this solution is filtered and the filtrate is concentrated under reduced pressure to obtain a crude product.

A small amount of hydroquinone and anhydrous ferric chloride are added to this crude product, and the crude product is purified by distillation under reduced pressure. As a result,
The new compound (I) is obtained as a pale yellow liquid.

The boiling point of this compound (I) at a pressure of 1 Torr is 96 to 98 ° C.
Met.

The compound (I) synthesized as described above was identified by IR spectrum and ¹ H-NMR, respectively.

According to the measurement results of IR spectrum, absorption of the ester at a wavenumber of 1725 cm ^-1 is absorption of carboxyl group at a wavenumber of 1700 cm ^-1 is absorption of alkenes were observed respectively at a wavenumber 1630,1320,1295cm ^-1.

In addition, as a result of measuring the ¹ H-NMR spectrum by dissolving the compound (I) in CDCl ₃ , the chemical shift value (δ) of ¹ H
Table 1 shows the functional groups that are considered to contain protons that give the shift value.

... Then, the compound (I) synthesized as described above is reacted with thionyl chloride in the presence of pyridine to synthesize methacryloyloxypropionyl chloride (hereinafter sometimes abbreviated as compound (II)). The procedure was described in. The following equation (2) is the reaction equation.

5.27 g (0.033 mol) of compound (I) was dissolved in 50 ml of anhydrous THF (tetrahydrofuran), and 11.9 g (0.1 mol) of this was dissolved.
Of thionyl chloride (SOCl ₂ ) and 5.27 g (0.067 mol) of pyridine are added. This mixture is heated and reacted at a temperature of 50 ° C. for 1 hour, and thionyl chloride and THF in the final reaction are distilled off under reduced pressure. Further distillation of the residue under reduced pressure gave a new compound (I
I) is obtained as a pale yellow liquid.

The boiling point of this compound (II) at a pressure of 1 Torr is 75 to 76 ° C.
Met.

The compound (II) synthesized as described above was identified by IR spectrum and ¹ H-NMR, respectively.

According to the measurement results of IR spectrum, absorption of the carbonyl group of the acid chloride at a wavenumber of 1800 cm ^-1 is absorption of ester wavenumber 1725 cm ^-1 is absorption of alkenes was observed respectively in the wavenumber 1630,1320,1295Cm ^-1 It was

The compound (II) was dissolved in CDCl ₃ and the ¹ H-NMR spectrum was measured. As a result, the ¹ H chemical shift value (δ)
Table 2 shows the functional groups that are considered to contain protons that give the shift value.

... Next, in addition to the compound (II) synthesized as described above, DL-α- which is an example of the compound represented by the general formula described above
A compound having a phospholipid-like structure of Example 1 was prepared by reacting phosphatidylethanolamine dibalmitoyl (compound represented by (III) in the formula (3) below) in the presence of pyridine by the procedure described below. Is 2-
(Methacryloyloxy) ethyldipalmitoyl-DL-
α-Phosphatidylethanolamide (a compound represented by (IV) in the following formula (3)) is produced. The following equation (3) is the reaction equation. The DL-α-phosphatidylethanolamine dipalmitoyl (III) was manufactured by Sigma (product number P3275).

First, in a three-necked flask equipped with a stirrer, a calcium chloride tube, and a dropping funnel, 0.20 g (0.29 mmol) of DL-α was added.
-Phosphatidyl ethanolamine dipalmitoyl (II
I) and 0.11 g (1.45 mmol) pyridine dissolved in 40 ml chloroform are added. Next, this flask was cooled to 0 ° C. in an ice bath, and while stirring the solution in the flask, 0.204 g (1.16 mmo) was added to this solution through a dropping funnel.
The compound (II) of l) is slowly added dropwise over 1 hour. Further, this solution is reacted at 0 ° C. for 6 hours and then at 20 ° C. for 12 hours. Then the reaction mixture is added with 20 ml of water and then extracted with 80 ml of chloroform. The extracted chloroform solution is dehydrated with sodium sulfate and then filtered, and the filtrate is concentrated under reduced pressure. The residue was dissolved in 0.3 ml of chloroform, and 120 ml of acetone was added thereto, and pale yellow crystals appeared. After leaving it in the freezer for 20 hours as it is, the precipitated pale yellow crystals are collected and washed with 20 ml of acetone. In this way a crude product is obtained.

The crude product is dissolved in chlorobenzene and purified by reprecipitation with acetone. As a result, the compound (IV) having a structure similar to the natural phospholipid of Example 1 is obtained.

This compound (IV) exhibited a liquid crystal phase in the temperature range of 42 to 139 ° C.

The compound (IV) of Example 1 synthesized as described above was identified by IR spectrum and ¹ H-NMR.

According to the measurement result of IR spectrum, the wave number is 2925,2850,1460
absorption of the methylene groups in cm ^-1 is absorption of ester wavenumber 1730 cm ^-1 is absorption of amide group at a wave number of 1655 cm ^-1, wave number 1630,1
Absorption of alkene 320,1295Cm ^-1 is absorption of phosphoryl group at a wavenumber of 1255cm ^-1 is absorption Hosuhorireto the wave number 1070 cm ^-1 was observed, respectively.

In addition, the compound (IV) of Example ¹ was dissolved in CDCl ₃ to give ¹ H-
As a result of measuring the NMR spectrum, Table 3 shows the chemical shift value (δ) of ¹ H and the functional group which is considered to contain a proton giving the shift value.

<Explanation of Polymer of Compound of Example 1> Next, a polymer of the compound of Example 1 is prepared by polymerizing the compound (IV) having a structure similar to the natural phospholipid of Example 1 produced as described above as described below. Was synthesized. As the polymerization initiator, 2,2'-azoisobutyronitrile (hereinafter abbreviated as AIBN) was used.

First, 0.21 g of the compound (IV) of Example 1 and 0.82 mg of AIB
Dissolve N in 5 ml chlorobenzene and place this solution in a glass tube. Next, the inside of the glass tube is replaced with nitrogen, and then the glass tube is sealed under reduced pressure. After that, 80 this glass tube
Shake for 24 hours at a temperature of ° C. Next, when the substance in the glass tube was taken out and added to acetone, the polymer of the compound of Example 1 was precipitated. Next, this precipitate is collected, washed with acetone, further dissolved in chlorobenzene, and reprecipitated with acetone for purification.

The structural formula of the polymer of the compound of Example 1 is shown below (wherein R ₁ and R ₂ in the formula are both (CH ₂ ) ₁₄ CH ₃ ).

The polymer of the compound of Example 1 was a light brown powder and softened at a temperature of 145 ° C.

Identification of the polymer of the compound of Example 1 synthesized as described above was carried out by IR spectrum and viscosity.

According to the IR spectrum measurement results, the compound (I
It was found that the absorption of the alkenes with wave numbers of 1320,1295 cm ^-1 observed in V) disappeared in the polymer, and the other absorption peaks did not change before and after the polymerization. From this, it can be seen that the polymerization reaction is normally performed.

The polymer of the compound of Example 1 was soluble in chlorobenzene, chloroform and hot ethanol,
It was insoluble in acetone and ether.

The intrinsic viscosity of the 0-dichlorobenzene solution of the polymer of the compound of Example 1 at 25 ° C was 0.23dl / g.

Example 2 Next, the compound (II) (methacryloyloxypropionyl chloride), which was an intermediate for producing the compound having a natural phospholipid-like structure of Example 1, and the compound represented by the general formula described above. Of L-α-Sephalin (a substance represented by (V) in the following reaction formula (4)), which is an example of the above, and has a natural phospholipid-like structure of Example 2 (hereinafter, the compound of Example 2). Methacryloyloxyethyl cephalinamide (a compound represented by (VI) in the following reaction formula (4)) is produced, and a polymer of the compound of Example 2 is produced. .

<Description of Compound of Example 2> First, the compound of Example 2 is produced as described below. The following equation (4) is the reaction equation. L-α
-Sepharin (V) is made by Sigma (product number P426
4) was used. This is extracted from the cerebrum of sheep and is represented by (V) in the reaction formula (4). However, in reality, R ₃ and R ₄ are a mixture of ethanolamine phosphatides in which each of R ₃ and R ₄ is an alkyl group having 12 to 18 carbon atoms or an alkenyl group having 12 to 18 carbon atoms.

First, in a three-necked flask equipped with a stirrer, a calcium chloride tube, and a dropping funnel, 2 g of L-α-sephalin (V) and 0.91 g (11.5 mmol) of pyridine were dissolved in 50 ml of chloroform. Put in. Next, the flask was cooled to 0 ° C. in an ice bath, and 2.04 g (11.6 m) of the solution in the flask was stirred into the solution through a dropping funnel.
(mol) of compound (II) is slowly added dropwise over 1 hour. Further, this solution is reacted at 0 ° C. for 6 hours and then at 20 ° C. for 12 hours. Then 50 ml of water is added to the reaction mixture and then extracted with 150 ml of chloroform. The extracted chloroform solution is dehydrated with sodium sulfate and then filtered, and the filtrate is concentrated under reduced pressure. The residue was dissolved in 0.5 ml of chloroform, and 200 ml of acetone was added thereto, and pale yellow crystals appeared. After leaving it in the freezer for 20 hours as it is, the precipitated pale yellow crystals are collected and washed with 20 ml of acetone. In this way a crude product is obtained.

The crude product is dissolved in chlorobenzene and purified by reprecipitation with acetone. As a result, the compound (VI) having a structure similar to the natural phospholipid of Example 2 is obtained.

The compound (VI) of Example 2 exhibited a liquid crystal phase in the temperature range of 32 to 125 ° C.

The compound (VI) of Example 2 synthesized as described above was identified by IR spectrum and ¹ H-NMR, respectively.

According to the measurement result of IR spectrum, the wave number is 2925,2850,1460
absorption of the methylene groups in cm ^-1 is absorption of ester wavenumber 1730 cm ^-1 is absorption of amide group at a wave number of 1655 cm ^-1, wave number 1630,1
Absorption of alkene 320,1295Cm ^-1 is absorption of phosphoryl group at a wavenumber of 1255cm ^-1 is absorption Hosuhorireto the wave number 1070 cm ^-1 was observed, respectively.

In addition, the compound (VI) of Example ₂ was dissolved in CDCl ₃ to give ¹ H-
As a result of measuring the NMR spectrum, Table 4 shows the chemical shift value (δ) of ¹ H and the functional group which is considered to contain a proton which gives the shift value.

<Explanation of Polymer of Compound of Example 2> Next, the polymer of the compound of Example 2 was prepared by polymerizing the compound (VI) having a structure similar to the natural phospholipid of Example 2 prepared as described above as described below. Was synthesized. As the polymerization initiator, AIBN was used as in Example 1.

5 ml of 0.21 g of the compound of Example 2 (VI) and 0.82 mg of AIBN
Dissolved in chlorobenzene and placed in a glass tube. Next, the inside of the glass tube is replaced with nitrogen, and then the glass tube is sealed under reduced pressure. Then, the glass tube is shaken at a temperature of 80 ° C. for 24 hours. Next, when the substance in the glass tube was taken out and added to acetone, the polymer of the compound of Example 2 was precipitated. Next, this precipitate is collected, washed with acetone, further dissolved in chlorobenzene, and reprecipitated with acetone for purification.

The structural formula of the polymer of the compound of Example 2 is shown below (provided that each of R ₃ and R ₄ in the formula represents an alkyl group having 12 to 18 carbon atoms or an alkenyl having 12 to 18 carbon atoms). In addition,
The polymer of the compound of this Example 2 had L-α as the starting material.
-The compound of Example 2 synthesized from cephalin (V) (V
Since I) is used, R ₃ for each monomer of this polymer,
R ₄ is not necessarily the same.

The polymer of the compound of Example 2 was a light brown powder and softened at a temperature of 145 ° C.

Identification of the polymer of the compound of Example 2 synthesized as described above was performed by IR spectrum and viscosity, respectively.

According to the IR spectrum measurement results, the absorption of the alkene having a wave number of 1320,1295 cm ^-1 observed in the compound (V) before the polymerization disappeared in the polymer, and other absorption peaks did not change before and after the polymerization. I understood. From this, it can be seen that the polymerization reaction is normally performed.

Further, the polymer of the compound of Example 2 was soluble in chlorobenzene, chloroform and hot ethanol,
It was insoluble in acetone and ether.

The intrinsic viscosity of the chlorobenzene solution of the polymer of the compound of Example 2 at 25 ° C was 0.19dl / g.

(Effect of the invention) As is clear from the above description, the compound having a natural phospholipid-like structure, which is the first invention of this application, has a methacrylic acid ester type polymerizable functional group,
The polymerization reaction easily takes place under normal polymerization conditions using a polymerization initiator such as AIBN. The compound of the first invention can, of course, give a polymer by irradiation with high energy rays such as γ rays, X rays, electron rays, and ultraviolet rays.

Further, the polymer of the compound having a natural phospholipid-like structure, which is the second invention of this application, has a property not found in conventional natural phospholipids, that is, because it is a polymer, it is extremely easy to form a membrane. Yes, and the resulting membrane is much stronger than natural phospholipids. Therefore, it can be used in a wide range of fields such as the humidity sensor, gas sensor, ion permeable membrane, artificial organ, cell fusion, enzyme immobilization, biosensor, and artificial cultivation described above, and its industrial value is very large.

Further, the method for producing a compound having a natural phospholipid-like structure of the third invention of the present application is directed to a compound having a natural phospholipid-like structure which can be easily polymerized not only by radiation polymerization but also by ordinary radical polymerization. , Commercially available DL-α
-Phosphatidylethanolamine dipalmitoyl or L-α-cephalin can be easily produced by using a few reaction steps. Therefore, the compound having the above-mentioned natural phospholipid-like structure having excellent properties can be provided economically and economically on an industrial scale.

Claims (3)

[Claims] 1. The following general formula (In the formula, each of R ₁ and R ₂ represents an alkyl group having 12 to 20 carbon atoms or an alkenyl group having 12 to 20 carbon atoms.) A compound having a natural phospholipid-like structure. 2. The following general formula (In the formula, each of R ₁ and R ₂ represents an alkyl group having 12 to 20 carbon atoms or an alkenyl group having 12 to 20 carbon atoms. Further, n represents the degree of polymerization and is a value within the range of 100 to 500. ) A polymer of a compound having a natural phospholipid-like structure represented by: 3. The following general formula (In the formula, each of R ₁ and R ₂ represents an alkyl group having 12 to 20 carbon atoms or an alkenyl group having 12 to 20 carbon atoms.) In producing a compound having a natural phospholipid-like structure, A step of reacting methacryloyl and propionic acid in the presence of sodium hydroxide to synthesize methacryloxypropionic acid, and a step of reacting the methacryloylpropionic acid with thionyl chloride in the presence of pyridine to synthesize methacryloyloxypropionyl chloride. The methacryloyloxypropionyl chloride having the following general formula
(Wherein each of R ₁ and R ₂ represents an alkyl group having 12 to 20 carbon atoms or an alkenyl group having 12 to 20 carbon atoms) in the presence of pyridine. A method for producing a compound having a characteristic natural phospholipid-like structure.