WO2020184797A1 - Eco-friendly method for preparing unnatural amino acid-based antibacterial and antibiotic preservative - Google Patents

Abstract

The present invention relates to: an eco-friendly method capable of synthesizing an unnatural amino acid-based antibacterial and antibiotic preservative, which has excellent antibacterial and antibiotic performance, in a high yield and with excellent processing efficiency; and a mass production system therefor. According to the present invention, a unnatural amino acid-based antibacterial and antibiotic preservative, which has excellent antibacterial and antibiotic performance, can be mass synthesized in a high yield and with excellent processing efficiency by an eco-friendly method, and thus the present invention can be used in various fields of the eco-friendly material industry such as that of cosmetic preservatives, food preservatives, cosmetic materials, biomedicines, functional amino acids, natural antibiotic alternatives, agrifood water preservatives, acaricides and the like such as preservatives or antibacterial and sterilizing agents.

Description

Eco-friendly manufacturing method of antibacterial antibiotic preservatives based on non-natural amino acids

The present invention relates to an antibacterial antibiotic preservative based on a non-natural amino acid, and more particularly, a method for synthesizing an antibacterial antibiotic preservative based on a non-natural amino acid with excellent antibacterial and antibiotic performance in an eco-friendly method, high yield, and excellent process efficiency. And a mass production system thereof.

Preservatives and antibiotics are currently in a situation where the development of a formulation having a new mechanism is required due to the problem due to resistance and the advent of super bacteria, and attempts to solve the problem are being made using candidate substances having various antibacterial activities.

LAE (ethyl lauroyl alginate) to be produced in the present invention can be applied to wet tissues containing moisture (natural pulp, cotton, tencel fabric, natural fiber fabric, nonwoven fabric, synthetic fiber fabric, blended fiber fabric, etc.), and It can also be applied to foods.

In addition, quaternary ammonium-based cetylpyridinium chloride (CPC), which is currently the most widely used as a preservative in the past, increases the amount of foam generated over time, so it is necessary to use a defoaming agent. It indicates various problems such as process inefficiency. In addition, there are various problems such as a phenomenon vulnerable to microbial contamination. In addition, several bacteria are proliferated during the manufacturing process of the product. Among them, Burkholderia sp. has a problem in that it can be infected by manufacturing facilities. For the above reasons, it is urgent to develop a new preservative that significantly reduces or completely replaces the amount of quaternary ammonium-based materials such as cetylpyridinium chloride.

With the development of a new preservative that significantly reduces or completely replaces the amount of cetylpyridinium chloride, the amount of creambazole, which has been commonly used as a preservative component at home and abroad, is increasing. Climbazole, which is recently used as a preservative, is a white crystalline powder and is known to have a lower irritation or risk than other ingredients as it is rated 0 in the EWG Skin Dip grade, an environmental research nonprofit in the United States. In addition, creambazole is known to have an exclusive and sustained sterilizing effect against ovarian nasal fungi and trichophytic bacteria, but it is insoluble in water and is a potentially toxic harmful substance.

In addition, Gram-negative bacillus is a major causative agent of nosocomial infection, and in recent years, its resistance range and degree are becoming more severe, making it more difficult to select empirical antibiotics. In particular, Gram-negative bacillus isolated in Korea has a remarkably higher resistance to antibiotics that are commonly used compared to foreign countries, and recently, resistance to Gram-negative bacillus is also expanding.

According to the manufacturing method of LAE (ethyllauroyl alginate) known to date, most of the manufacturing methods are made under conditions that are heavily influenced by low temperature reaction conditions and pH, and the process cost efficiency such as the amount of reagent used for synthesis and synthesis time is low. There is a low problem.

Therefore, the present invention provides a method for synthesizing an antimicrobial antibiotic preservative based on a non-natural amino acid that is biodegradable and has excellent antibacterial and antimicrobial performance in an eco-friendly method and with a high yield and excellent process efficiency in order to solve such problems. It aims to solve the problem, the problem of super bacteria, and the problem of mass production.

That is, the present invention is to provide a synthetic equipment used as a method of obtaining a preservative having an antimicrobial antiseptic effect from an amino acid and a synthetic method using the same.

Accordingly, the present invention provides a method for synthesizing an antibacterial antibiotic preservative based on a non-natural amino acid represented by the following [Chemical Formula 1], including the following steps in order to solve the above problem.

(i) obtaining an intermediate represented by [Chemical Formula 2] using an amino acid as a starting material according to the following [Scheme A],

[Scheme A]

[Formula 2]

(ii) obtaining [Chemical Formula 1] from [Chemical Formula 2] according to the following [Scheme B],

[Scheme B]

[Formula 1]

In addition, the present invention provides a method for synthesizing an antibacterial antibiotic preservative based on a non-natural amino acid represented by the following [Chemical Formula 3], including the following steps in order to solve the above problem.

(iii) obtaining an intermediate represented by [Chemical Formula 4] using an amino acid derivative as a starting material in the presence of a salt according to the following [Scheme C],

[Scheme C]

[Formula 4]

(iv) obtaining [Chemical Formula 3] from [Chemical Formula 4] according to the following [Scheme D],

[Scheme D]

[Formula 3]

Detailed descriptions of [Scheme A], [Scheme B], [Scheme C], and [Scheme D], and specific structures of [Chemical Formula 1] to [Chemical Formula 4] will be described later.

In addition, the present invention provides a system capable of synthesizing and producing a large amount of antimicrobial antibiotic preservatives based on non-natural amino acids according to this manufacturing method.

According to the present invention, antimicrobial antimicrobial preservatives based on non-natural amino acids with excellent antibacterial and antibiotic properties can be mass-synthesized in an eco-friendly method, high yield, and excellent process efficiency, and thus cosmetic preservatives, food preservatives, cosmetics It can be used in various fields of the eco-friendly material industry such as raw materials, biopharmaceuticals, functional amino acids, natural antibiotic substitutes, agro-food water preservatives, and mite acaricides.

1 is a system diagram of a synthetic reaction apparatus for preparing an amino acid-based antibacterial antibiotic preservative according to the present invention.

2 is an image of a liquid preservative composition comprising an amino acid-based antibacterial antibiotic preservative according to the present invention.

3 to 5 are antibacterial test results for the liquid preservative composition according to an embodiment of the present invention, Figure 3 is E. coli, Figure 4 is Staphylococcus aureus, Figure 5 is a test result for Candida.

6 is a preservation test result of a liquid preservative composition according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail.

The present invention relates to a method for preparing an amino acid-based antimicrobial antibiotic preservative having a structure as shown in [Chemical Formula 1], and a method for synthesizing an antibacterial antibiotic preservative based on a non-natural amino acid represented by the following [Formula 1] including the following steps It is about.

(i) obtaining an intermediate represented by [Chemical Formula 2] using an amino acid as a starting material according to the following [Scheme A],

[Scheme A]

[Formula 2]

(ii) obtaining [Chemical Formula 1] from [Chemical Formula 2] according to the following [Scheme B],

[Scheme B]

[Formula 1]

In the above formula, R ₁ and R ₂ are the same as or different from each other and are each independently selected from an alkyl group having 1 to 20 carbon atoms, an alkenyl group having 2 to 20 carbon atoms, and an alkynyl group having 2 to 20 carbon atoms, and R is a structure of 20 amino acids It is a side chain that satisfies.

The step (i) is characterized in that it comprises the step of adding sodium hydroxide (NaOH) and alcohol to the aqueous amino acid solution and stirring while adding alkanal (R ₁ COH) chloride while stirring.

The step (ii) is characterized in that it comprises the step of reacting hydrochloric acid gas while stirring the [Chemical Formula 2] and alcohol, and the hydrochloric acid gas is a hydrochloric acid produced by reacting sulfuric acid and sodium chloride by designing a reactor configuration characteristically. It is characterized by using a gas.

In addition, the present invention relates to a method for preparing an amino acid-based antimicrobial antibiotic preservative having a structure as shown in [Chemical Formula 3] below, and including the following steps, of a non-natural amino acid-based antibacterial antibiotic preservative represented by the following [Formula 3] It relates to the synthesis method.

(iii) obtaining an intermediate represented by [Chemical Formula 4] using an amino acid derivative as a starting material in the presence of a salt according to the following [Scheme C],

[Scheme C]

[Formula 4]

(iv) obtaining [Chemical Formula 3] from [Chemical Formula 4] according to the following [Scheme D],

[Scheme D]

[Formula 3]

In the above formula, R ₁ and R ₂ are the same as or different from each other and are each independently selected from an alkyl group having 1 to 20 carbon atoms, an alkenyl group having 2 to 20 carbon atoms, and an alkynyl group having 2 to 20 carbon atoms, and R is a structure of 20 amino acids Is a side chain satisfying, and Y is selected from HF, HCl, HBr and HI as a salt.

In reflection of these characteristics, the present invention provides a system capable of mass-synthesizing antibacterial antibiotic preservatives based on non-natural amino acids represented by the above [Chemical Formula 1].

The synthesis system according to the present invention is largely composed of a hydrochloric acid gas generating unit and a synthetic reaction unit of an antibacterial antibiotic preservative based on a non-natural amino acid.

That is, a sulfuric acid solution storage tank containing a sulfuric acid solution, a sodium chloride storage tank containing sodium chloride, a first pipe supplying a compressed gas to the sulfuric acid solution storage tank, the sulfuric acid solution storage tank and the sodium chloride storage tank communicate with each other, and the compressed gas Including a second pipe for transporting the sulfuric acid solution to the sodium chloride storage tank when supplied, a first on-off valve mounted on the first pipe, and a second on-off valve mounted on the second pipe, the sodium chloride storage tank Hydrochloric acid gas generator for generating hydrochloric acid gas from; And,

A synthetic reaction tank for synthesizing an antibacterial antibiotic preservative based on a non-natural amino acid by receiving the hydrochloric acid gas, a third pipe for transporting the hydrochloric acid gas by communicating the sodium chloride storage tank and the synthetic reaction tank, and a fourth pipe in communication with the third pipe It consists of a pipe, and a synthetic reaction unit including a three-way valve mounted on a connection portion between the third pipe and the fourth pipe.

Hereinafter, the present invention will be described in more detail through examples and experimental examples. These Examples and Experimental Examples are only intended to specifically illustrate the present invention, and the scope of the present invention is not limited by these Examples and Experimental Examples.

Synthesis of N-α-Lauroyl- D- Arginine Ethyl ester ( D- LAE) corresponding to an embodiment of the present invention according to the following synthesis example was synthesized through a reaction apparatus configured as follows.

Synthesis Example 1. Synthesis of N-α-Lauroyl- D -Arginine Ethyl ester ( D- LAE)

(1) D- arginine and 50 mL of distilled water were added and dissolved in a 250 mL, two neck round bottom flask according to the following [Reaction Scheme 1], and then NaOH 9.2 g was added. Then, 50 mL of t-Butyl alcohol was added and stirred. After lowering the temperature to 1 to 4 °C, 13.2 mL of lauroyl chloride was added dropwise and stirred rapidly for 6 hours. The white solid was filtered, washed three times with 100 mL of 85% t-Butyl alcohol aqueous solution, and filtered.

[Scheme 1]

(2) According to the following [Reaction Scheme 2], add 10 g of N-lauroyl- D -Arginine and 100 mL of ethanol to a 250 mL, 3-necks round bottom flask, and stir at room temperature, and the Argon gas cock of the gas generator of FIG. 1 Open, open the sulfuric acid injection cock 1/3, and add 4.5 mL of sulfuric acid to 9.9 g of sodium chloride. Open the discharge cock of the reaction vessel, open the hydrochloric acid injection cock, and inject the generated hydrochloric acid gas. After all injections, raise the hydrochloric acid gas injection pipe so that it does not come into contact with the reactants, close the hydrochloric acid gas injection cock, close the discharge cock, and raise the temperature to 80 ℃. Excess residual hydrochloric acid gas is heated and stirred for 6 hours after returning the cock to the discharge port connected to the water bottle.

[Scheme 2]

Synthesis Example 2. Synthesis of N-α-Lauroyl- D -Arginine Ethyl ester ( D- LAE)

(1) According to the following [Reaction Scheme 3], add 50 g of D- Arginine and 500 mL of ethanol to a 1000 mL, 3-necks round bottom flask, followed by stirring at room temperature. Open the argon gas cock of the gas generator, open the sulfuric acid injection cock, and slowly drop sulfuric acid onto the sodium chloride solid. Open the discharge cock of the reaction vessel, open the hydrochloric acid injection cock, and inject the generated hydrochloric acid gas. After all injections, raise the hydrochloric acid gas injection pipe so that it does not come into contact with the reactants, close the hydrochloric acid gas injection cock, close the discharge cock, and raise the temperature to 80 ℃. Excess residual hydrochloric acid gas is heated and stirred for 6 hours after returning the cock to the outlet connected to the water bottle.

[Scheme 3]

(2) According to the following [Reaction Scheme 4], 50 g of D- Arginine ethyl ester dihydrochloride and 250 mL of distilled water were added and dissolved in a 1000 mL, two neck round bottom flask, and then 250 mL of t-Butyl alcohol was added and stirred. After lowering the temperature to 1 to 4 °C, 13.2 mL of Lauroyl chloride was added dropwise, and 30.7 g of NaOH was added, followed by rapid stirring for 6 hours. After filtering the white solid, it was washed three times with 500 mL of 5% hydrochloric acid (35% aq.) / 85% t-Butyl alcohol / 10% distilled water and filtered.

[Scheme 4]

N-α-Lauroyl- D -Arginine Ethyl ester ( D -LAE) (Yield 82%).

White solid, melting point 57° C., Exact MASS 384.3100 ESI-MS; m/z 385.3172 (m+H);

¹ H NMR: δH (CDCl ₃ ), 0.89 [t, 3H, (CH ₃ alkyl chain)], 1.29 [m, 18H, (9CH ₂ , alkyl chain)], 1.62-1.78 [m, 4H, (-CH ₂ -CH ₂ -CONH-), (CH ₂ -CH ₂ -NH-C(=NH)-NH ₂ )], 2.03 [s, 1H, (-NH-C(=NH)-NH ₂ )], 2.29 [t, 2H, (-CH ₂ CONH-)], 3.24-3.42 [m, 2H, (CH ₂ -NH-C(=NH)-NH ₂ )]], 4.2 [m, 2H, (-OCH ₂ -CH ₃ )], 4.44 [1H, (-NH-CHCOO-)], 7.19 [4H, (-NH-C(=NH)-NH ₂ )], 7.91 [1H, (-NH-CH-COO )]

Synthesis Example 3

Compounds	MW (g/mol)	Mole	EQ	Amount	Density(g/cm3)
L-Arginine HCl	210.66	0.01	One	21g
H 2 SO 4	98.08	0.025	2.5	25 g (14 mL)	1.84
CaCl 2	110.98	0.0275	2.75	31g
EtOH				200 mL
Lauroyl chloride	218.76	0.0096	0.96	21 g (22 mL)	0.946
H 2 O				200 mL
NaHCO 3	87.01	0.06	6	52 g
EA				200 mL

According to the following [Scheme 5], 21 g of L-Arginine HCl and 31 g of CaCl ₂ were added to 100 mL of the two necks round bottom flask, 200 mL of EtOH was added and stirred for 10 minutes, 14 mL of H ₂ SO4 was added, followed by 12 hours at 100°C. During reflux. After cooling to room temperature, it is concentrated under reduced pressure to remove EtOH, and 200 mL of distilled water is added to the reaction precipitate, followed by stirring and dispersion. After adding 52 g of NaHCO ₃ slowly, 200 mL of ethyl acetate (EA) was added and stirred, 22 mL of Lauroyl chloride was slowly added, followed by stirring at room temperature for 3 hours. Stir the reaction mixture, adjust the pH between 4 and 5 with concentrated hydrochloric acid, add 200 mL of EA, separate the organic layer, remove moisture with MgSO ₄ , filter, and concentrate under reduced pressure to form white powder Etyhl lauroyl arginate HCl (95%, 40 g) is obtained.

[Scheme 5]

Examples and Experimental Examples

(1) After composition of the N-α-Lauroyl- D -Arginine Ethyl ester ( D- LAE) synthesized according to Synthesis Examples 1 and 2 with a liquid sample L preservative as shown in FIG. 2, respectively, E. coli and Staphylococcus aureus And the results of the antibacterial test for each of Candida bacteria according to the test method "KCL-FIR-1002:2018" are shown in [Table 2] below.

Test Items		Test Methods	Test result			Environment
			Initial concentration (CFU/mL)	After 24 hours (CFU/mL)	Bacterial reduction rate
Antibacterial test E. coli	BLANK	KCL-FIR-1002:2018	1.8×10 4	1.2×10 5	-	37±0.2(℃)
	Sample L		1.8×10 4	<10	99.9
Staphylococcus aureus	BLANK		3.5×10 4	5.9×10 4	-
	Sample L		3.5×10 4	<10	99.9
Candida	BLANK		1.9×10 4	1.4×10 5	-
	Sample L		1.9×10 4	<10	99.9

Results photos for the [Table 2] are shown in Figs. 3 to 5, respectively, and in [Table 2] and Figs. 3 to 5, N-α-Lauroyl- D- Arginine Ethyl ester synthesized according to the present invention ( It can be seen that the antibacterial performance of D- LAE) is very excellent.

(2) After composition of the N-α-Lauroyl- D- Arginine Ethyl ester ( D- LAE) synthesized according to Synthesis Examples 1 and 2 with a liquid sample L preservative as shown in FIG. 2, 80 g of cotton in a sealed container After diluting 100 g of water and 1 g of the preservative, it was left on an ondol floor (2 days) and on a desk for 5 days for a week to conduct a preservation test, and the results are shown in Table 3 and FIG. 6 below.

division	Sodium benzoate	Sample L	VP3	Grapefruit Seed Extract
color	Little change	Color change appears	Color change appears	Severe color change
smell	slightly	Less	Severe	Severe
mold	A little from the inside	Begins to appear	Appear from the inside	Severe
Starch concentration	Less	slightly	plenty	plenty

As shown in [Table 3] and Fig. 6 (① sodium benzoate/② sample L/③ VP3/④ grapefruit seed extract), N-α-Lauroyl- D -Arginine Ethyl ester ( D- LAE) according to the present invention Compared to sodium benzoate, the effect of color change, mold development, and starch concentration is somewhat lower, but it can be confirmed that it is rather superior in terms of odor, and both show excellent effects compared to VP3 and grapefruit seed extract, so even in the preservation test. High effect can be confirmed.

According to the present invention, antimicrobial antimicrobial preservatives based on non-natural amino acids with excellent antibacterial and antibiotic properties can be mass-synthesized in an eco-friendly method, high yield, and excellent process efficiency, and thus cosmetic preservatives, food preservatives, cosmetics It can be used in various fields of the eco-friendly material industry such as raw materials, biopharmaceuticals, functional amino acids, natural antibiotic substitutes, agro-food water preservatives, and mite acaricides.

Claims (5)

1. A method for synthesizing an antibacterial antibiotic preservative based on a non-natural amino acid represented by the following [Chemical Formula 1] including the following steps:

   (i) obtaining an intermediate represented by [Chemical Formula 2] using an amino acid as a starting material according to the following [Scheme A],

   [Scheme A]

   

   [Formula 2]

   (ii) obtaining [Chemical Formula 1] from [Chemical Formula 2] according to the following [Scheme B],

   [Scheme B]

   

   [Formula 1]

   In the above formula, R ¹ and R ² are the same as or different from each other and are each independently selected from an alkyl group having 1 to 20 carbon atoms, an alkenyl group having 2 to 20 carbon atoms, and an alkynyl group having 2 to 20 carbon atoms, and R is a structure of 20 amino acids It is a side chain that satisfies.
2. A method for synthesizing an antibacterial antibiotic preservative based on a non-natural amino acid represented by the following [Chemical Formula 3] including the following steps:

   (iii) obtaining an intermediate represented by [Chemical Formula 4] using an amino acid derivative as a starting material in the presence of a salt according to the following [Scheme C],

   [Scheme C]

   

   [Formula 4]

   (iv) obtaining [Chemical Formula 3] from [Chemical Formula 4] according to the following [Scheme D],

   [Scheme D]

   

   [Formula 3]

   In the above formula, R ₁ and R ₂ are the same as or different from each other and are each independently selected from an alkyl group having 1 to 20 carbon atoms, an alkenyl group having 2 to 20 carbon atoms, and an alkynyl group having 2 to 20 carbon atoms, and R is a structure of 20 amino acids Is a side chain satisfying, and Y is selected from HF, HCl, HBr and HI as a salt.
3. The method of claim 1,

   The (ii) step is a method for synthesizing an antibacterial antibiotic preservative based on a non-natural amino acid, characterized in that it comprises the step of reacting hydrochloric acid gas while stirring the [Chemical Formula 2] with alcohol.
4. The method of claim 3,

   The hydrochloric acid gas is a method of synthesizing an antimicrobial antibiotic preservative based on a non-natural amino acid, characterized in that using hydrochloric acid gas generated by the reaction of sulfuric acid and sodium chloride.
5. As a synthetic system of antibacterial antibiotic preservatives based on non-natural amino acids represented by [Chemical Formula 1] according to claim 1,

   The synthesis system,

   A sulfuric acid solution reservoir containing a sulfuric acid solution, a sodium chloride reservoir containing sodium chloride, a first pipe supplying a compressed gas to the sulfuric acid solution reservoir, the sulfuric acid solution reservoir and the sodium chloride reservoir are communicated, and the compressed gas is supplied. Hydrochloric acid in the sodium chloride storage tank, including a second pipe for transporting the sulfuric acid solution to the sodium chloride storage tank, a first on/off valve mounted on the first pipe, and a second on/off valve mounted on the second pipe A hydrochloric acid gas generator generating gas; And

   A synthetic reaction tank receiving the hydrochloric acid gas and synthesizing an antibacterial antibiotic preservative based on a non-natural amino acid, a third pipe for transporting the hydrochloric acid gas by communicating the sodium chloride storage tank and the synthesis reaction tank, and a fourth pipe communicating with the third pipe A synthetic reaction unit including a pipe, and a three-way valve mounted on a connection portion between the third pipe and the fourth pipe;

   Synthesis system of antibacterial antibiotic preservatives based on non-natural amino acids comprising a.

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