TWI342313B - Vinyl alcohol-vinyl hydroxyl benzoate copolymer and composite film comprising the same - Google Patents

Description

1342313, IX, invention: [Technical Field] The present invention relates to a copolymer, in particular to a vinyl alcohol-hydroxybenzoic acid vinyl ester copolymer and a composite film comprising the same . [Prior Art] The bioactive functional group is polymerized to form a continuous repeating unit polymer, or is connected to the polymer main chain by other reaction means to form a material design or enhance utility of different conventional use patterns, and has been developed. Into a specialized technology® field. The development of polymers with antibacterial activity has been published in related literatures. It can be divided into three major systems according to the way of activity. One is the bactericidal activity of the polymer body. Such representative examples are polycationic compounds (such as A polymer containing a biguanide suspension base or poly(vinyl salicylic acids) or the like. The second is to use the polymer as a matrix to decompose and release the active functional group at an appropriate timing, such as ciprofloxacin-polycarprolactone condensation polymer or polyvinyl alcohol-(peptide link) Sub)-PVA-(peptide linker)-gentamicin copolymer. The third is to introduce the bacteriostatic agent into the polymer matrix through material technology to achieve controlled release. There are quite a few examples of this type, such as nylon (nylon) as the gauze material, polystyrene (polystyrene) as the matrix or A drug designed to bind an antibiotic to the stomach for a long period of time with a 2-hydroxypropyl methylcellulose in combination with a foaming design. It is an excellent candidate for hydroxybenzoic acid to bond a functional group having antibacterial activity into a long-chain molecule to make it a wholly active copolymer. For example, propyl para-methyl benzyl benzoate ( propyl / 7-hydroxybenzoate 0806-A20988TWF (N2); david 5 1342313 (propyl paraben)) has an inhibitory ability against most of the genus, and the esterification bonds of the above derivatives are It occurs on the carboxylic acid group of para-hydroxybenzoic acid in order to make it active or even enhanced. In addition, Albertsson et al. used a similar structure of 2-hydroxy benzoic acid, a salicylic acid, to esterify with a polyacrylic acid to form a pendant functional group. The antibiotic activity was found to be lost after the base polymer. When the hydroxyl group and the carboxylic acid group of 2-hydroxybenzoic acid are simultaneously retained and repolymerized with ethylene at the fourth carbon to form a polymer, the activity is even larger than that of the original monomer molecule. By comparing the formation of para-hydroxybenzoic acid esters and the manner in which 2-hydroxy alum acid is bonded to a polymeric chain (polyacrylic acid or polyethylene), it is believed that the hydroxyl groups on the para-hydroxy alum acid are retained and allowed to It should have certain antibacterial activity when exposed to the outside of the molecule. As for the repeating monomer structure on the main chain of the esterified with a carboxylic acid group, if it is maintained at 4 carbon numbers, the antibacterial activity should be modified, and Dumitriu et al. have used naphthalene bonding to form naphthalene. A successful example of the formation of a copolymer of nalidixic acid and PVA, so polyvinyl alcohol (PVA) should be a suitable choice for the polymer backbone. SUMMARY OF THE INVENTION In view of the above, the present invention provides a vinyl alcohol-hydroxybenzoate copolymer having the following chemical formula (I): (CH2CH)m(CH2CH)n

OH (I) 0806-A20988TWF (N2): david 6 1342313 wherein m is 42 to 99, and η is 1 to 57. The present invention further provides a composite film comprising a gel film comprising the above-described vinyl alcohol-hydroxybenzoic acid vinyl ester copolymer. [Embodiment] The present invention provides a vinyl alcohol-hydroxybenzoate copolymer having the chemical formula (I):

In the formula (I), m may be 42 to 99, and η may be 1 to 57. The chemical formula (I) can be synthesized in the following manner. First, a mixed hydroxybenzoic acid and a solvent are placed in a reaction flask. The hydroxybenzoic acid may include an ortho, meta or para-substituted hydroxy citric acid or a hydroxy-cracked acid containing two or more hydroxy groups, and the solvent may be Ν-methyl-2-nitroglyoxime. (N-methyl-2-pyrrolidone, ΝΜΡ), Ν, Ν dimethyl dimethylformamide (DMF), tetrahydrofuran (THF) or acetone. Next, an activator is slowly added at room temperature to activate the base group of the benzoic acid. The activator may comprise a gasified thionyl chloride 'three gasified fill or five gasified scales. After the unreacted activator is removed, vaporized hydroxybenzimid is obtained. Next, the mixed polyvinyl alcohol and the solvent were placed in a reaction flask and the reaction was heated. The solvent used may include N-methyl-2-pyrrolidone (NMP), N,N-dimethylformamide (DMF), and four. 0806-A20988TWF (N2); david 7 1342313 Hydrofurfuran (THF) or acetone (acetone). After cooling, the above-mentioned gasified hydroxybenzimid reaction solution is slowly added, and esterification reaction is carried out at different reaction temperatures (for example, room temperature, 65 degrees Celsius and 100 degrees Celsius), and the reaction can be obtained. Vinyl alcohol-hydroxybenzoate copolymers of different graft ratios (esterification rates). The grafting reaction of the product obtained by the esterification reaction at different reaction temperatures of the present invention has a graft ratio of vinyl alcohol to hydroxy quesinate of from about 40 to 99%. The copolymer of the present invention can be used as an antibacterial agent against Staphylococcus aureus (• Minimum inhibitory concentration of Siap/ijv/ococcws (Minimum Bacterial)

Inhibition Concentration (MIC) is generally between 0.05 and 0.25%, and the minimum inhibitory concentration against Candida albicans is between 0.5 and 1.0%. Further, the copolymer may also function as a conductive polymer. The present invention further provides a composite film comprising a gel film comprising the above-described vinyl alcohol-hydroxybenzoate copolymer. Next, the production of the composite film of the present invention will be described. First, a vinyl alcohol-hydroxybenzoate copolymer solution and a gel polymer are mixed. The copolymer solution further includes, for example, N-methyl-2-pyrrolidone (NMP), N,N-dimercaptocarbamide (Ν, Ν-dimethylformamide, DMF). ), a solvent of tetrahydrofuran (THF) or acetone, and the gel polymer may be a monolithic gel polymer. After the two are uniformly mixed and dried, the residual solvent is removed to obtain the composite film. The features and advantages of the present invention are further illustrated by the following examples. EXAMPLES Example 1 0806-A20988TWF(N2); david 8 1342313 Synthesis of vinyl alcohol-para-hydroxybenzoic acid copolymer (CH2CH)m (CH2(j:

H)n OH Ο I c=o

OH synthesis step: (1) Synthesis of vaporized para-hydroxy alum

First, mix 10 g of para-hydroxy benzoic acid (equivalent to 〇〇 724 m〇le) with 40 ml of N-methyl-2-nitrosudolone (NMP) in a round flat-bottomed glass bottle and stir. Dissolved. At the same time, a reflow device is assembled, and a glass outlet tube containing anhydrous calcium carbide is added to the interface of the condenser to remove air and moisture. Then, in the process of (4), 4.8 ml of di-sulfurized sulphur (equivalent to 0.0657 mo丨e) was added. The reaction molar ratio of benzoic acid to di-sulfurized sulphur was 1. 0.9. Continue to react under the dish! hour. After the reaction, the reflux device was removed, and the reaction flask was transferred to a vacuum drying oven and heated under a pressure of 15 cm of a mercury column (C and a temperature of 40 C for 24 hours to remove unreacted gasified sulfurized sulfur and sulfur dioxide. 'At this time, care should be taken to keep the operating environment dry. 0806-A20988TWF(N2) :david 1342313 (2) Synthesis of vinyl alcohol-para-hydroxybenzoic acid vinyl ester copolymer

(CHsCHLi^CH^ Ο OH c=o

OH

A suitable amount of polyvinyl alcohol and nmp were mixed in a round flat-bottomed glass bottle, heated in an oil bath to about 120 ° C to dissolve completely and taken out for cooling. At the same time, a reflow device is assembled and a glass tube containing anhydrous calcium carbide is added to the condenser port to remove moisture from the air. During the stirring process, the gasification para-hydroxybenzamide reaction solution containing the molar number of the vinyl alcohol monomer is slowly added, and the esterification reaction is carried out at three reaction temperatures (including room temperature, 65 ° C and 100 Torr, respectively). Finally, the vinyl alcohol-para-hydroxy hydroxybenzoate copolymer of the present invention was obtained, and the average esterification rates obtained at the respective reaction temperatures were 41.6% (room temperature), 97.4±2.2% (65. 0), and 97.3, respectively. Soil 〇.6% (1〇〇.〇. In the process of synthesizing the ethylene-para-hydroxybenzoic acid copolymer, the temperature is obviously a key factor for increasing the acetification rate, and also affects the esterification reaction to saturation. Time of time 'If the heating temperature is insufficient, even after a long period of reaction, the final esterification rate is relatively low. Therefore, 'increasing the temperature should allow the reactants to have enough energy to overcome the energy barrier of the activated complex and help the reaction proceed. For example, increasing the temperature can overcome the molecular repulsion generated when the graft side chain is formed by 0806-A20988TWF(N2); david 10 1342313. If the temperature continues to increase to 100 〇c, the final purified sample will be relatively hard, but hydrolyzed. Rate analysis The rate of conversion can still be as high as 97.3% ± 0.6%. Example 2 Minimum inhibitory concentration (MIC) determination The MIC referred to herein refers to the culture solution containing 2 X 1 4 cells (ceus) per ml, and finally complete The minimum concentration of bacteriostatic components required to inhibit colony growth. The MIC test first requires the cultivation of three strains of single strain, that is, the glass beads of the frozen strain attached to Escherichia coli ATCC10536 are first added, and about 4 ml of sterilized trypsin is added. Soy agar medium (TSB) was shaken to dissolve the adherent cells into a turbid liquid, and the suspension was taken up with a 1 ML-capacity transfer line and streaked onto a nutrient agar disc culture dish to 37 Incubate in °C for 24 hours, so that a single colony is obtained at the proximal end of the scribing line. Pick the single colony into the test tube containing 10 ml of nutrient br〇th and mix evenly. After 37 C was cultured for another 20 hours, the concentration of the bacterial solution in the obtained test tube was 108 ± 1004 cells/mL (1σ, the number of measurements n=3), and then diluted to a concentration of 106 cells/mL with sterile distilled water for the antibacterial test. Staphylococcus aureus (10) like (10) ~ ATCC6538P) Two glass beads of frozen strain, the same as the previous E. coli operation mode, but with sterile distilled water instead of tryptic soy agar medium (TSB), the concentration of the obtained liquid in the test tube was 108 soil 10O7cells/mL (la, n=3) And diluted with sterile distilled water to a concentration of H) 6 CellS / mL. Finally, the attached Candida albicans (c (10) heart like ATCC10231) cold; 4 strains of glass beads of the eastern strain, add about 4 ml of sterilized steaming water, spin to make the attached bacteria dissolve into a turbid liquid, to transfer the capacity of the bacteria Circle the suspension 0806-A20988TWF (N2): david 1342313 liquid 'striped on the yeast and mold medium (YM agar) disc culture dish, incubate at 25 C constant temperature for 48 hours, so that the end of the line A single colony was obtained, and the single colony was picked by a transfer fungus, mixed in a test tube containing 1 ml of nutrient liquid medium, and then cultured again at 25 ° C for 48 hours, and the concentration of the obtained liquid in the test tube was 106 ± l. 0-2 cells/mL (1σ, n=3). After the culture of the strain is completed, the MIC measurement is started. First, the para-benzoic acid and the esterified polymer (test group) are separately prepared into a series of concentrations of the test liquid, including 1.0%, 0.5%, and 0.25%. And 0.05%. Since the acetated polymer is hardly soluble in water, the 1.0% esterified polymer is prepared by heating with propylene glycol and then diluting with water. The concentration of propylene glycol is 10% (ν/ν)·> The group consists of solvents corresponding to different concentrations of the test solution, including 10%, 5%, 2.5%, 0.5% (v/v) aqueous propylene glycol solution. After that, three kinds of bacterial liquids of 〇·丨 ml and concentration of 1〇6 cells/mL· were collected by using a micropipette, and 4.9 ml of the above-mentioned various concentrations of the esterified polymer test liquid and the para-hydroxybenzoic acid test were respectively added. Mix the liquid and the blank separately. Next, incubated at 25 C for 24 hours to carry out a bacteriostatic reaction test, and take a test solution with 1 〇μ [capacity of the transfer zone, streak the dish, where the E. coli and S. aureus culture dish Nutrient agar was used as the medium, and Candida albicans used yeast and mold medium (YM agar). The lines of the three media were repeated for each tube. The culture of the E. coli and S. aureus test groups was 37. c was inverted for 24 hours, and the Candida albicans test group was at 25. (: Inverted culture for 48 hours, and finally the cultured blood was taken out to observe the colony growth. The MIC was judged by the colony observed by the colony counter magnifying glass, and the three culture media in which the individual bacteriostatic agents were repeatedly cultured on a single strain were determined. The minimum bacteriostatic concentration required for aseptic growth is the MIC. Here, the room temperature is varnished with 0806-A20988TWF (N2); david 12 1342313 copolymer, and finally the copolymer is obtained against Staphylococcus aureus and large intestine. The hops of Bacillus licheniformis and Candida albicans were (4) 5%, G 25%, and Q 5% (w/v), respectively, and the MIC of the para-basic benzoic acid to the above species was respectively: N., 〇, 1.0% ( Wv)”x The results of this test show that the MIC of the self-derived polymer seems to have a more bacteriostatic effect than the monomolecular para-benzoic acid against Staphylococcus aureus and white bacteria. Example 3 Production of composite film First , to make a gel material, the steps are as follows, mix 5 ml of tetraethoxysilane (2 cc, 95% ethanol in a fresh ML beaker, and magnetic (four) device (four), too rich and add people H) ML, _ N salt At room temperature, for another 4 hours, it can be obtained. The gelatin coffee network (four) state of the silicone network polymer β and the above-mentioned stone sol are mixed to form a film of various solutions respectively (1) mixed with i grams of polyethylene Alcohol with 1 () ml of hydrazine, Ν _: methyl methamine in the European oil bath to soften, disperse ', then add water to dissolve, and finally dilute to ML with water. (2) Preparation containing 〇 gram pair a solution of benzoic acid with iq ml hydrazine, hydrazine-dimethyl ketoamine. (3) mixed gram of copolymer product with h ml of hydrazine, hydrazine dimethyl dimethyl carbamide & 13G ° C oil The bath was heated and dissolved to prepare a -copolymer stock solution. Next, two composite films were prepared, the first containing 5% (w/w) of the acetated polymer 'the second containing 5% (w/ w) the para-position of phenylic acid, the third does not contain any bacteriostatic agent / 7 t as the step is to add the appropriate amount of the above solution to the human gel (si ^ a yogel), and then The mixture was plated in a hard-made small box and dried under a vacuum of 120 C and Α g for 16 hours to remove the residual solution 0806-A20988TWF (N2); david 1342313 agent especially 疋 (5) N'N-dimethylformamide at the point of the point can form a dry composite film, which is taken out in a flat form and measured for average thickness by a Waltmam Mass. AMES 214-25 Thick Sound Meter. Example 4 Antibacterial Benefit Evaluation of Composite Film The antibacterial test of the composite film was prepared by diluting the above-mentioned concentration of 1〇6ceUs/mL of Escherichia coli 3, gold f color, and Staphylococcus aureus standard solution in a nutrient liquid medium to a concentration of 10, 10, 10 cells. /mL of the dilution, and each of the two 1 ml of the dilution was transferred to a Petri dish t (90 mm in diameter) containing about 15 ml of nutrient bad fat medium, which was evenly distributed on the surface of the medium by shaking. After coagulation, the various films of 15 χ 1 5 coffee 2 size _ were placed on the bacteria-containing surface with a sterile recording, and two sets of culture dishes were obtained. The group was Escherichia coli, and the other group was golden yellow (four) cocci, each group. Each of the culture dishes containing three different bacterial numbers (103, 1〇4, 1〇5 cells) was flattened with an f# membrane at equidistant positions. Finally, the film-coated culture dish was placed in a box and inverted for 24 hours. - The antibacterial benefit evaluation of the composite film of the present invention is carried out by Gram staining. The reason for using this method is that any surviving bacterial cells on the film will grow more colonies if they are transplanted by a similar single culture method. It is easy to be troublesome in judging the antibacterial effect, and Gram stain can stain the test sample with the control group (ie, the film containing no bacteriostatic agent) and the comparison group (ie containing 5% para-base benzene). The formic acid film was used as a clear control to judge the antibacterial effect. Remove the cultured film with a sterile record. After cutting, place the bacterial contact side of the film face up and lay it on the slide. Drip 1 to 2 drops of sterilized steaming water onto the film for 2 to 3 minutes, then quickly pass The flame is fixed to the film, and then a few drops of purple oxalate crystal solution (〇xa]ate crystal vi〇] et s-) is finished with the film 0806-A20988TWF (N2); david 1342313 full coverage 'after standing for 1 minute, The film was washed with sterilized distilled water, and then a solution of a total amount of iodine solution (lugol-PVP Iodine complex solution) was applied to the film to completely collapse the solution. After standing for 1 minute, the film was washed with sterilized distilled water. The continuously used ethanol-acetone mixture (50:50, v/v) was decolorized to remove any pigment and then added dropwise to a few drops of safranin (safranin s〇luti〇n) and completely covered with the film. After being colored for 1 minute, it was washed with sterilized distilled water, air-dried, magnified by a microscope to 6 倍 times to observe the staining condition, and the image was recorded by a digital camera of a charge coupling device. The antibacterial benefit of the 矽 gel composite film is evaluated by Gram staining method such as Gram-positive bacteria Staphylococcus aureus (the film should be blue when the brother is present, and the Gram-negative bacteria such as Escherichia coli The film should be red. The results showed that in the group of 丨〇3 Staphylococcus aureus colonies implanted in the culture, the film containing 5% of the copolymer synthesized at room temperature showed a slight negative reaction (the film was red) "It shows that the bacteria can not grow effectively. When the antibacterial agent and the other polymers form a composite film, the active functional groups are basically limited to the cavity of the polymer structure, and the effective space for the activity is reduced, that is, free. The volume of the free volume is reduced, and in order to reduce such limitations and to properly evaluate the resistance of a high esterification rate polymer which is generally insoluble in a solvent, the present invention is selected to have a porous cavity and has affinity for many microorganisms. The bismuth gel is used as the film substrate. Although the invention has been disclosed above in the preferred embodiments, it is not intended to limit the invention. Anyone skilled in the art will not Within the spirit and scope of the present invention, there may be some modifications and simplifications. Therefore, the scope of the present invention is defined in the appended claims. 0806-A20988TWF(N2);david 15 1342313 • · [Simple diagram Description] - 〇4 *«, [Description of main component symbols] fe 〇«»»>

0806-A20988TWF(N2);david 16

Claims (1)

1342313 Amendment No. 94123147 Revision date: 99.7.2 Patent application scope: ' 1. A vinyl alcohol-hydroxybenzoic acid vinyl ester copolymerization formula (I) (CH2CH) m(CH2CH)n Ο OH I c-o OH (I) Wherein m is 42 to 99 and n is 1 to 57. The copolymer is an antibacterial agent, and the Minimum Bacterial Inhibition Concentration (MIC) for Staphylococcus aureus/ococcws is generally between 0.05 and 0.25. %, the minimum Bacterial Inhibition Concentration (MIC) for Candida albicans a/Mca/w) is generally between 0.5 and 1.0%. 2. A composite film comprising: a sol-gel film comprising an ethylenic-hydroxybenzoate copolymer as described in claim 1 of the patent application. 3. The composite film of claim 2, wherein the gel film is a silica sol-gel.