KR20190090443A - Antimicrobial agents with metal organic frameworks - Google Patents

Abstract

The present invention relates to an antimicrobial agent comprising a metal-organic framework compound, wherein the metal-organic framework has excellent antimicrobial activity against bacteria and is very safe.

Description

Antimicrobial agents with metal organic frameworks

The present invention relates to an antimicrobial agent comprising a metal-organic framework.

The Metal-Organic Framework (MOF) has been used in the last few decades for a wide range of applications such as energy-related gas storage (H ₂ and CH ₄ ), CO ₂ capture / separation, hydrocarbon separation, catalysis and proton conduction. Has attracted a lot of attention in the field. In recent years, the high porosity and regular structural properties have led to the expansion of metal-organic frameworks for use in biology and medicine.

Meanwhile, while multidrug resistant bacteria are formed to threaten the development of traditional drugs, transition metal ions and metal nanoparticles including Cu, Zn, Co, and Ag are attracting attention as alternatives to new antimicrobial agents that affect bacterial cells. Metal nanoparticles exhibit excellent antibacterial activity due to inherent properties such as nanometer effect and high surface area / volume ratio. However, excess metal ions reached in the form of nanoparticles are expected to be harmful to normal tissue as well as bacteria.

To solve this problem, many researchers attempted to capture metal ions by coordinating them with organic ligands. In this regard, Korean Patent Laid-Open Publication No. 2009-0046888 et al. Discloses a method for producing a metal-organic framework including copper, but there is no report on the possibility of activity of the antimicrobial agent.

Republic of Korea Patent Publication 2009-0046888

CrystEngComm, 2015, 17, 456

Accordingly, the present invention was to provide an antimicrobial agent comprising a metal-organic framework.

In order to achieve the above object, the present invention provides an antimicrobial agent comprising a metal-organic framework of the formula (1).

[Formula 1]

[M ₂ (L1) ₂ (μ-L2)] x (H ₂ O)

Wherein M is Cu, Zn or Co,

L1 is a hydrocarbon group having two carboxylate groups,

L 2 is a bis (4-pyridyl) compound or a derivative thereof

x is a number from 1 to 10.

In the present invention, a metal (II) salt compound, a hydrocarbon compound having two carboxylate groups, and a bis (pyridyl) compound are mixed with water, and then dissolved. It provides a method for preparing a metal-organic framework compound of the formula (1) characterized in that the reaction for one day.

[Formula 1]

[M ₂ (L1) ₂ (μ-L2)] x (H ₂ O)

Wherein M is Cu, Zn or Co,

L1 is a hydrocarbon group having two carboxylate groups,

L 2 is a bis (4-pyridyl) compound or a derivative thereof

x is a number from 1 to 10.

The present invention is effective in providing an antimicrobial agent including a metal-organic framework having excellent antimicrobial activity. The present invention has the effect of providing a method for producing a metal-organic framework compound under relatively mild conditions using water as a solvent. Metal-organic framework compounds of the present invention include Escherichia coli, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Pleudomonas aeruginosa, Klebsiella pneumoniae, and MRSA (Methicillin Staphyloc) Aureus) has the advantage of excellent antimicrobial activity and very safe against bacteria such as.

1 illustrates the structural formula of the metal-organic framework compound of the present invention.
2 is a crystal structure of a metal-organic framework compound of the present invention. [Cu ₂ (Glu) ₂ (bpy)] (a), [Cu ₂ (Glu) ₂ (bpa)] (b), [Cu ₂ (Glu) ₂ (bpe)] each containing a water molecule (c) , And [Cu ₂ (Glu) ₂ (bpp)] (d).
3 shows the antimicrobial activity of the control group ((a)) and [Cu ₂ (Glu) ₂ (bpa)]. (H ₂ O) at 0.002 ((b)) and 0.01% ((c)) against Staphylococcus aureus. The effect results are shown.
Figure 4 shows the results of the antimicrobial effect of the control and [Cu ₂ (Glu) ₂ (bpa)]. (H ₂ O) for E. coli.
Figure 5 shows the antimicrobial effect of the control and [Cu ₂ (Glu) ₂ (bpa)]. (H ₂ O) against Pseudomonas aeruginosa.
6 shows the antimicrobial effect of the control group and [Cu ₂ (Glu) ₂ (bpa)]. (H ₂ O) against pneumococcal.
Figure 7 shows the results of the antimicrobial effect of the control and [Cu ₂ (Glu) ₂ (bpa)]. (H ₂ O) against MRSA bacteria.

The present invention provides an antimicrobial agent comprising a porous metal-organic framework. In particular, the metal-organic framework of the present invention has been designed in consideration of the inherent properties of metals and organic ligands such as oxidation water, counter ions, coordination modes, ligand size and crosslinking properties.

The present invention provides an antimicrobial agent comprising a metal-organic framework compound of Formula 1 below.

[Formula 1]

[M ₂ (L1) ₂ (μ-L2)] x (H ₂ O)

Wherein M is Cu, Zn or Co,

L1 is a hydrocarbon group having two carboxylate groups,

L 2 is a bis (4-pyridyl) compound or a derivative thereof

x is a number from 1 to 10.

The metal-organic framework is harmless to the human body because the water molecules are crystallized.

The metal of the metal-organic framework compound of the present invention is copper, zinc or cobalt. Copper, zinc or cobalt in the present invention can increase the antibacterial effect.

L1 of the present invention is a hydrocarbon group containing two carboxylate groups. L1 is preferably the following Chemical Formula 2.

(2)

^- OOC-R-COO ^-

In the above formula, R is a bond, a linear or branched hydrocarbon group having 1 to 7 carbon atoms which may have a single bond or a double bond.

L1 is more preferably malonat, succinate, fumarate, glutarate, adipate or muconate.

The metal of the metal-organic framework compound of the present invention is a L2 ligand in which bis (4-pyridyl) or a derivative thereof is bound. The bis (4-pyridyl) or a derivative thereof is preferably represented by the following Chemical Formula 3.

(3)

In the above formula

A is a C1-C7 linear or branched hydrocarbon group which may be a bond, a single bond, or a double bond.

Bis (4-pyridyl) is more preferably 4,4'-bipyridine (bpy), 1,2-bis (4-pyridyl) ethane (1,2-bis (4) -pyridyl) ethane, bpa), 1,2-bis (4-pyridyl) ethene (1,2-bis (4-pyridyl) ethene, bpe) or 1,3-bis (4-pyridyl) propane (1 , 3-bis (4-pyridyl) propane, bpp).

Hereinafter, the structure of the compound of the present invention will be described as an example of the Cu-organic framework compound. However, the present invention is not limited to the following compounds.

The Cu-glutarate metal-organic frameworks of the present invention were all crystallized in the monoclinic C2 / c space group. Single crystal X-ray studies have shown that all of the metal-organic frameworks of the present invention form a two-dimensional (2D) sheet shape, including paddle-wheel Cu ₂ heteronuclear units connected by glutarate. The two-dimensional sheets are linked by bipyridyl ligands (L2) to form a three-dimensional (3D) framework (see FIGS. 1 and 2). In FIG. 2, in the formula [Cu ₂ (Glu) ₂ (μ-L2)] x (H ₂ O), the 3D framework when L2 is bpy is represented by (a), and the 3D framework when bpa is represented by (b). , the 3D framework in the case of bpe is shown in (c), and the 3D framework in the case of bpp is shown in (d). The number of water molecules as a solvent was calculated from elemental analysis and the like. The coordination structure of each Cu (II) ion in the four metal-organic framework compounds is in the form of a square pyramid consisting of four equiatorial carboxylate O atoms and axial pyridyl N atoms.

Solvate molecules in the metal-organic framework compounds of the invention are captured in one-dimensional (1D) channels. The metal-organic frameworks of the present invention each have a pore volume of 10-40% by volume, preferably 15-35% by volume, based on PLATON analysis. The metal-organic framework compounds of the present invention all contain well-defined 1D channels and have very similar pore shapes with suitable pore size and pore volume. It is believed that these pores and pore volumes have an effect of improving antibacterial activity.

The metal-organic framework compound of the formula 1 of the present invention is not limited but is preferably prepared by the following method.

[Formula 1]

[Cu2 (L1) 2 (μ-L2)] x (H ₂ O)

Wherein M is Cu, Zn or Co,

L1 is a hydrocarbon group having two carboxylate groups,

L 2 is a bis (4-pyridyl) compound or a derivative thereof

x is a number from 1 to 10.

In the present invention, a metal (II) salt compound, a hydrocarbon compound having two carboxylate groups, and a pyridyl compound are mixed with water and then dissolved. After the reaction in a high pressure vessel at 25 ~ 100 ℃ for 12 hours to 10 days. After cooling the reaction solution, the resulting crystals are recovered.

The metal (II) salt compound may be a nitrate, sulfate, carbonate or chloride salt of the metal.

The hydrocarbon compound having two carboxylate groups is preferably HOOCRCOOH (R is a straight or branched chain hydrocarbon group having 1 to 7 carbon atoms which may be a bond, or may have a single bond or a double bond) or its sodium salt, calcium salt or Potassium salt.

When mixing with water, ultrasonic waves may be used to increase solubility, and NaOH aqueous solution may be added if necessary.

The metal-organic framework compounds of the present invention have antimicrobial activity.

The compounds of the present invention are particularly preferred, such as Escherichia coli, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, and MRSA (Methicillin-Recusistant Staus) such as Staphylococcus. Excellent antibacterial activity against bacteria

Hereinafter, the present invention will be described in detail through one embodiment. However, the present invention is not limited to the following examples.

Production example. [Cu ₂ (Glu) ₂ (L2)] x (H ₂ O) Manufacture

_{1. [Cu 2 (Glu) 2} (bpy)] · 3 (H 2 O)

Cu (NO ₃ ) ₂ · 3H ₂ O (0.193 g, 0.8 mmol), glutaric acid (0.317 g, 2.4 mmol) and 4,4'-bipyridine (0.0625 g, 2.4 mmol) in distilled water (20 mL) The solution containing was sonicated and placed in a high pressure vessel lined with Teflon. The vessel was placed in an oven at 80 ° C. for 48 hours. After cooling to room temperature, the cyan block crystals were recovered by filtration, washed with distilled water and dried in air overnight. The yield was 0.028 g. _{[Cu 2 (C 5 H 6} O 2) 2 (C 10 H 8 N 2)] · 3 (H 2 O) (MW = 597.51)

2. [Cu ₂ (Glu) ₂ (bpa)] · 3 (H ₂ O)

Cu (NO ₃ ) ₂ · 3H ₂ O (0.48 g, 2.0 mmol), glutaric acid (0.26 g, 2.0 mmol) and 1,2-bis in distilled water (200 mL) containing 1.0 M NaOH (2 mL) (4-pyridyl) ethane (0.18 g, 1.0 mmol) was added and placed in a high pressure vessel lined with Teflon. The vessel was placed in an oven at 80 ° C. for 48 hours. After cooling to room temperature, the turquoise crystal needles were recovered by filtration, washed with distilled water and dried in air overnight. The yield was 0.40 g. _{[Cu 2 (C 5 H 6} O 2) 2 (C 12 H 12 N 2)] · 3 (H 2 O) (MW = 622.05)

_{3. [Cu 2 (Glu) 2} (bpe)] · 6 (H 2 O)

In distilled water (200 mL) containing 1.0 M NaOH (2 mL), Cu (NO ₃ ) ₂ · 3H ₂ O (0.48 g, 2.0 mmol), glutaric acid (0.26 g, 2.0 mmol) and 1,2-bis (4-pyridyl) ethylene (0.18 g, 1.0 mmol) was added and placed in a high pressure vessel lined with Teflon. The vessel was placed in an oven at 80 ° C. for 48 hours. After cooling to room temperature, the turquoise crystal needles were recovered by filtration, washed with distilled water and dried in air overnight. The yield was 0.46 g. [[Cu ₂ (C ₅ H ₆ O ₂ ) ₂ (C ₁₂ H ₁₀ N ₂ )] · 6 (H ₂ O) (MW = 678.10)

4. [Cu ₂ (Glu) ₂ (bpp)]. 6 (H ₂ O)

Cu (NO ₃ ) ₂ .3H ₂ O (0.0624 g, 0.25 mmol), glutaric acid (0.033 g, 0.25 mmol), 1,3-bis (4-pyridyl) propane (0.050 g, 0.25 mmol) and urea 0.052 g, 0.87 mmol) was left at room temperature for 7 days. Turquoise crystals were recovered by filtration, washed with distilled water and dried in air overnight. Yield 0.033 g. [Cu ₂ (C ₅ H ₆ O ₂ ) ₂ (C ₁₃ H ₁₄ N ₂ )] · 6 (H ₂ O) (MW = 693.64)

The structural analysis results of the compound are shown in Table 1 below.

One 2 3 4 Empirical formula C ₂₀ H ₂₆ Cu ₂ N ₂ O ₁₁ C ₂₂ H ₁₂ Cu ₂ N ₂ O ₉ C ₂₂ H ₂₆ Cu ₂ N ₂ O ₁₁ C ₂₃ H ₂₆ Cu ₂ N ₂ O ₉ Formula weight 597.51 575.42 612.53 601.54 Temperature (K) 296 (2) 223 (2) 296 (2) 296 (2) Wavelength (?) 0.71073 Å 0.71073 Å 0.71073 Å 0.71073 Å Space group C 2 / c C 2 / c C 2 / c C 2 / c

5. Preparation of [Co ₂ (Glu) ₂ (bpa) ₂ ]

Co (NO ₃ ) ₂ .6H ₂ O (0.232 g, 0.8 mmol), glutaric acid (0.317 g, 2.4 mmol) and 1,2-bis (4-pyridyl) ethane (0.0625 g, in DMF (20 mL) 2.4 mmol) was sonicated and placed in a high pressure vessel lined with Teflon. The vessel was placed in an oven at 130 ° C. for 48 hours. After cooling to room temperature, the crystals were recovered by filtration, washed with DMF and dried in air overnight. The yield was 0.028 g. [Co ₂ (C ₅ H ₆ O ₂ ) ₂ (C ₁₂ H ₁₂ N ₂ ) ₂ ] (MW = 723.34)

6. Preparation of [Zn (Glu) (bpe)] 2 (H ₂ O)

1,2-bis (4-pyridyl) ethylene (0.0376) on a solution of Zn (NO ₃ ) ₂ .6H ₂ O (0.0304 g, 0.1 mmol) and glutaric acid (0.0133 g, 0.1 mmol) in 4 mL H ₂ O. g, 0.2 mmol) and carefully lift up the 4 mL acetonitrile solution. Decisions can be obtained after two weeks of standing. Yield 0.0287 g. [Zn (C ₅ H ₆ O ₂ ) ₂ (C ₁₂ H ₁₀ N ₂ ) ₂ ] · 2 (H ₂ O) (MW = 413.73

Examples. Antibacterial test

The concentration of [Cu ₂ (Glu) ₂ (bpa)]. (H ₂ O) was prepared at 0.002% and 0.01%, and the antibacterial effect was measured by the following procedure. The same was true for the control group to determine the antimicrobial effect.

1) Sample preparation: Samples should be prepared according to the concentration to be tested.

2) Put the prepared sample in the Erlenmeyer flask.

3) Sterilize by using sterilizer at 121 ℃ for 15 minutes

4) In a sterile Erlenmeyer flask, add 99 mL of physiological saline (NaCl 8.5%) (the amount of physiological saline can be changed depending on the sample concentration) and 1 mL of cultured test strain. (Total volume 100mL, test strain is cultured in Nutrient liquid medium)

5) Shake incubation for 24 hours at 37 ℃.

6) Collect a certain amount from the shaken solution and spread it on agar medium.

7) Measure the number of bacteria.

Example 1.Antibacterial effect against Staphylococcus aureus

The antibacterial test for Staphylococcus aureus was carried out and the results are shown in Table 2 below. In addition, the concentration of control group ((a)) and [Cu ₂ (Glu) ₂ (bpa)]. (H ₂ O) of 0.002 ((b)) and 0.01% ((c)) against Staphylococcus aureus is also shown in FIG. The antimicrobial effect results are shown.

Test Items
Test result Test Methods Test environment Initial concentration
(CFU / mL) Concentration after 24 hours
(CFU / mL) Cytotoxicity
(%) Antibacterial test by Staphylococcus aureus BLANK 1.9 x 10 ⁴ 8.2 x 10 ⁵ - KCL-FIR-1002: 2011 (37.0 ± 0.2) ℃ Cu ₂ (Glu) ₂ (bpa)
(0.002%) 1.9 x 10 ⁴ <10 99.9 Cu ₂ (Glu) ₂ (bpa)
(0.01%) 1.9 x 10 ⁴ <10 99.9

CFU: Colony Forming Unit

Inoculation Bacterial Concentration (CFU / mL): Staphylococcus Aureus: 1.9x10 ⁶

Use strain: Staphylococcus aureus ATCC 6538

Example 2 Antimicrobial Effects Against Escherichia Coli and Pseudomonas Aeruginosa

Antibacterial test for Escherichia coli and Pseudomonas aeruginosa was carried out and the results are shown in Table 3 below. In addition, Figure 4 shows the results of the antimicrobial effect of the control group and [Cu ₂ (Glu) ₂ (bpa)] · (H ₂ O) for E. coli. Figure 5 shows the antimicrobial effect of the control and [Cu ₂ (Glu) ₂ (bpa)]. (H ₂ O) against Pseudomonas aeruginosa.

Test Items
Test result Test Methods Test environment Initial concentration
(CFU / mL) Concentration after 24 hours
(CFU / mL) Cytotoxicity
(%) Antibacterial test by E. coli BLANK 3.0 x 10 ⁴ 8.2 x 10 ⁵ - KCL-FIR-1002: 2011 (37.0 ± 0.2) ℃ Cu ₂ (Glu) ₂ (bpa)
(0.002%) 3.0 x 10 ⁴ <10 99.9 Antibacterial test with Pseudomonas aeruginosa BLANK 3.2 x 10 ⁴ 7.4 x 10 ⁵ - Cu ₂ (Glu) ₂ (bpa)
(0.002%) 3.2 x 10 ⁴ <10 99.9

Inoculation Bacterial Concentration (CFU / mL): Escherichia Coli: 3.0x10 ⁶ ,, Pseudomonas Aeruginosa: 3.2x10 ⁶

Use strain: Escherichia coli ATCC 25922

Pseudomonas aeruginosa ATCC15442

Example 3 Antibacterial Effect against Pneumococcal and MRSA

The antibacterial test for pneumococci and MRSA bacteria was performed and the results are shown in Table 4 below. In addition, Figure 6 shows the results of the antimicrobial effect of the control and [Cu ₂ (Glu) ₂ (bpa)]. (H ₂ O) against pneumococci. Figure 7 shows the results of the antimicrobial effect of the control and [Cu ₂ (Glu) ₂ (bpa)] (H ₂ O) against MRSA bacteria.

Test Items
Test result Test Methods Test environment Initial concentration
(CFU / mL) Concentration after 24 hours
(CFU / mL) Cytotoxicity
(%) Antibacterial test by pneumococcal BLANK 2.0 x 10 ⁴ 3.0 x 10 ⁵ - KCL-FIR-1002: 2011 (37.0 ± 0.2) ℃ Cu ₂ (Glu) ₂ (bpa)
(0.002%) 2.0 x 10 ⁴ <10 99.9 Antimicrobial test by MRSA BLANK 2.3 x 10 ⁴ 8.1 x 10 ⁵ - Cu ₂ (Glu) ₂ (bpa)
(0.002%) 2.3 x 10 ⁴ <10 99.9

According to the antibacterial test results, it can be seen that the compound of the present invention has excellent antibacterial effect against various bacteria.

Claims (7)

An antimicrobial agent comprising a metal-organic framework of Formula 1
[Chemical Formula 1]
[M ₂ (L1) ₂ (μ-L2)] x (H ₂ O)
Wherein M is Cu, Zn or Co,
L1 is a hydrocarbon group having two carboxylate groups,
L 2 is bis (4-pyridyl) or a derivative thereof
x is a number from 1 to 10.
The antimicrobial agent of claim 1, wherein L1 is a group represented by the following Chemical Formula 2.
(2)
^- OOC-R-COO ^-
Wherein R is a bond, a straight or branched chain hydrocarbon group having 1 to 7 carbon atoms which may have a single bond or a double bond The antimicrobial agent according to claim 1, wherein L2 is a compound of Formula 3
(3)

In the above formula
A is a C1-C7 linear or branched hydrocarbon group which may be a bond, a single bond, or a double bond. The compound of claim 1, wherein L 2 is 4,4′-bipyridine (bpy), 1,2-bis (4-pyridyl) ethene (1,2-bis (4-pyridyl) ethene, bpe), 1,2-bis (4-pyridyl) ethane (1,2-bis (4-pyridyl) ethane, bpa) or 1,3-bis (4-pyridyl) propane (1,2-bis ( 4-pyridyl) propane, bpp)
A metal (II) salt compound, a hydrocarbon compound having two carboxylate groups, and a pyridyl compound are dissolved in water, and then reacted for 12 hours to 10 days at 25 to 100 ° C. in a high pressure vessel. Method for preparing metal-organic framework compound
[Chemical Formula 1]
[M ₂ (L1) ₂ (μ-L2)] x (H ₂ O)
Wherein M is Cu, Zn or Co,
L1 is a hydrocarbon group having two carboxylate groups,
L 2 is bis (4-pyridyl) or a derivative thereof
x is a number from 1 to 10. The method according to claim 5,
Metal (II) salt compound is a manufacturing method characterized in that the metal nitrate, sulfate, carbonate or chloride salt The method according to claim 5,
A hydrocarbon compound having two carboxylate groups is HOOCRCOOH (R is a bond, a straight or branched chain hydrocarbon group having 1 to 7 carbon atoms which may have a single bond or a double bond), or HOOCRCOOH (R is a bond, or a single bond. Method for producing a sodium salt, calcium salt or potassium salt of a linear or branched hydrocarbon group having 1 to 7 carbon atoms which may have a double bond

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