阅读说明：本技术 材料单体OGAn-CA及其制备方法与应用 (Material monomer OGAN-CA, and preparation method and application thereof ) 是由 夏中华 万蕊 于 2021-10-26 设计创作，主要内容包括：本发明涉及医学材料领域,具体而言,提供了一种材料单体OGA-(n)-CA及其制备方法与应用。材料单体OGA-(n)-CA为氰基丙烯酸酯官能团与聚乙醇酸片段的化学结合,既具有氰基丙烯酸酯类材料快速聚合固化的特征,又具有聚乙醇酸材料良好的生物相容性和生物降解性。(The invention relates to the field of medical materials, and particularly provides a material monomer OGA n -CA and a preparation method and application thereof. Material monomer OGA n CA is the chemical combination of cyanoacrylate function and polyglycolic acid segment, and has the characteristic of rapid polymerization and solidification of cyanoacrylate materials and good biocompatibility and biodegradability of polyglycolic acid materials.)

1. Material monomer OGA_n-CA characterized in that said OGA_n-structural formula I of CA is as follows:

wherein n is an integer of 1 to 10.

2. Monomeric OGA of material according to claim 1_nThe preparation method of the-CA is characterized in that the material monomer OGA is obtained by esterification reaction of a compound with a structural formula II and alpha-cyanoacrylate protected by anthracene and deprotection group reaction_n-CA，

Wherein n is an integer of 1 to 10.

3. A composition of matter, a method of making,characterized in that said composition comprises monomeric OGA of the material of claim 1_n-CA。

4. The composition of claim 3, wherein the composition is a monomeric OGA of material_n-a plurality of combinations in CA.

5. Composition according to claim 3 or 4, characterized in that it further comprises a pharmaceutically acceptable adjuvant selected from thickeners, stabilizers, thermal and/or photo initiators and accelerators to initiate cross-linking, colorants, plasticizers, preservatives, heat-dissipating agents, biocompatible agents and/or fibrous reinforcing materials.

6. The composition of claim 3 or 4, wherein the composition further comprises one or more biological agents or therapeutic agents.

7. Monomeric OGA of material according to claim 1_n-CA or a composition according to any one of claims 3 to 6 for use in medicine.

8. Use according to claim 7, characterized in that it comprises the preparation of medical adhesives or tissue engineering materials;

preferably, the medical adhesive comprises adhesives for wound adhesion, hemostasis, visceral and soft tissue wound closure, coverage, leakage stoppage, hard tissue fixation.

9. A polymer prepared by OGA of the material monomer of claim 1_n-CA or the composition according to any of claims 3 to 6, obtained by cross-linking copolymerization under the action of anions.

10. The polymer of claim 9, wherein the anion is-OH and/or-NH₂-OH and/or-NH₂Derived from tissues, body fluids,At least one of skin and blood.

Technical Field

The invention relates to the field of medical materials, in particular to a material monomer OGA_n-CA and a preparation method and application thereof.

Background

CA, named cyanoacrylate, was first synthesized by a German chemist in the last 50 th century, and its structure has strong electron withdrawing capacity of cyano group and ester group to promote the activity of olefinic double bond and fast polymerization in the presence of anion. Furthermore, cyanoacrylate has strong adhesive force, and the synthesized monomer may be compounded with stabilizer, plasticizer and other additives (such as tackifier) to improve biocompatibility, stability and clinical performance, so that it has become a medical adhesive commonly used in clinical in recent years, and researchers now use it for embolism varicose vein and closed cornea surgery in addition to the conventional functions of hemostasis, wound adhesion and sterilization.

The cyanoacrylate has important and wide application in hemostasis and wound adhesion, and has fast polymerization phase change characteristic to reach the effect of fast hemostasis and wound adhesion and to isolate outer germs, and liquid-solid phase change characteristic makes it easy to carry and use. However, cyanoacrylate materials also have some limitations at present, and researches show that although the toxicity of the long-chain derivatives is lower, the more complicated synthesis and manufacturing technology limits the industrial development and application to a certain extent; the biodegradation rate of cyanoacrylate also needs to be optimized, the long carbon chain polymer formed by polymerization phase change is slowly degraded under physiological conditions, and the molecular weight is often as high as hundreds of thousands or even millions of daltons; in addition, the adhesion is too high and not easy to remove, and the polymerization is too fast and not easy to control, which limits the further application of such materials. Thus, despite the great potential for the application of cyanoacrylate war wound dressings, there is a need for further improvements in their biocompatibility and biodegradability.

In view of the above, the present invention is particularly proposed.

Disclosure of Invention

The first purpose of the invention is to provide a material monomer OGA_n-CA。

The second purpose of the invention is to provide a material monomer OGA_n-a process for the preparation of CA.

A third object of the present invention is to provide a composition.

The fourth purpose of the invention is to provide a material monomer OGA_n-CA or use of a composition.

A fifth object of the present invention is to provide a polymer.

In order to achieve the above purpose of the present invention, the following technical solutions are adopted:

material monomer OGA_n-CA, said OGA_n-CA has the formula I：

Wherein n is an integer of 1 to 10.

Monomer OGA of the above material_nThe preparation method of the-CA comprises the steps of carrying out esterification reaction on a compound with a structural formula II and anthracene-protected alpha-cyanoacrylate, and then carrying out deprotection group reaction to obtain the material monomer OGA_n-CA，

Wherein n is an integer of 1 to 10.

A composition comprising monomer OGA of the above material_n-CA。

Further, the composition is monomer OGA_n-a plurality of combinations in CA.

Further, the composition further comprises a pharmaceutically acceptable adjuvant selected from thickeners, stabilizers, thermal and/or photo initiators and accelerators to initiate cross-linking, colorants, plasticizers, preservatives, heat dissipating agents, biocompatible agents and/or fiber reinforcing materials.

Further, the composition also includes one or more biological agents or therapeutic agents.

Monomer OGA of the above material_n-CA or the use of a composition in medicine.

Further, the use comprises preparing a medical adhesive or a tissue engineering material;

preferably, the medical adhesive comprises adhesives for wound adhesion, hemostasis, visceral and soft tissue wound closure, coverage, leakage stoppage, hard tissue fixation.

A polymer prepared by OGA of the monomer_n-CA or the composition is obtained by crosslinking copolymerization under the action of anions.

Further, the anion is-OH and/or-NH₂-OH and/or-NH₂Derived from at least one of tissue, body fluid, skin and blood.

Compared with the prior art, the invention has the beneficial effects that:

the invention takes cyanoacrylate structure as polymerizable group, chemically joins oligomeric glycolic acid segments (OGA) with different lengths in monomer molecule, obtains brand new polymerizable curing material monomer OGA_n-CA. Material monomer OGA_nThe CA monomer has small molecular weight and is liquid at normal temperature, and because of containing cyanoacrylate functional groups, olefinic bonds in the monomer can be subjected to rapid intermolecular polymerization when contacting a human body, and a liquid material is solidified to form a polymer film, so that the liquid material can rapidly cover and fill wounds to play the roles of physically protecting and blocking bacteria. Meanwhile, the obtained polymer has good biocompatibility and flexibility because of containing a large amount of OGA side chains, and more importantly, the OGA contains a plurality of ester bonds easy to hydrolyze, so that the polymer can be endowed with good degradation performance. The physical and chemical properties of the polymer can be further regulated and controlled by regulating the number of OGA repeating units, so that different practical application requirements are met. In addition, nuclear magnetic results show that the material monomer is successfully synthesized; the physical and chemical properties of the material are characterized by means of curing time, shear tensile strength, GPC (molecular exclusion chromatography), in-vitro degradation test and the like, and the results show that all the monomers of the material can be polymerized on the organism, and meanwhile, the material can generate effective adhesive force, and the polymer has low molecular weight and good degradation performance.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.

FIG. 1 is a monomer OGA of the material of example 2₃Nuclear magnetic characterization results of the polymerization phase transition of CA.

Detailed Description

Embodiments of the present invention will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer.

Unless otherwise defined, technical and scientific terms used herein have the same meaning as is familiar to those skilled in the art. In addition, any methods or materials similar or equivalent to those described herein can also be used in the present invention.

The invention provides a material monomer OGA_n-CA having the following structural formula I:

wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

Through the chemical combination of the cyanoacrylate function and the polyglycolic acid segment, the material system has the characteristic of rapid polymerization and solidification of cyanoacrylate materials, and has good biocompatibility and biodegradability of polyglycolic acid materials. Specifically, the material system can jointly generate adhesion capacity based on the engagement effect of the main chain of the polycyanoacrylate and the hydrogen bond and electrostatic effect of the OGA branched chain, and the purpose that the polymer firmly covers and protects the wound surface is achieved. Meanwhile, hydrogen bonds and electrostatic action are easy to be quickly damaged under the condition of water molecule infiltration, and the purpose of removing the water molecules as required can be realized. The polymer contains a large number of OGA branched chains, and has good biodegradability based on the terminal autocatalysis mechanism of OGA and the control of polymerization degree in the linear polymerization process.

Monomer OGA of the above material_nThe preparation method of-CA can be as follows: after the compound with the structural formula II and anthracene-protected alpha-cyanoacrylate undergo esterification reaction, the compound undergoes deprotection group reaction to obtain a material monomer OGA_n-CA，

Wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

Esterifying carboxyl on anthracene-protected alpha-cyanoacrylate with hydroxyl of compound of formula II to obtain intermediate reactant, deprotecting carboxyl and removing anthracene protecting group to obtain monomer OGA_n-CA。

For example, when n is 1, the material monomer OGA₁The synthetic route for-CA is as follows:

when n is 2, the material monomer OGA₂The synthetic route for-CA is as follows:

when n is 3, the material monomer OGA₃The synthetic route for-CA is as follows:

the invention provides a composition, which can comprise monomer OGA of the material provided by the invention_nAny one of-CA, and also in various combinations.

In a preferred embodiment, the composition further comprises a pharmaceutically acceptable adjuvant selected from thickeners, stabilizers, thermal and/or photo initiators and accelerators to initiate cross-linking, colorants, plasticizers, preservatives, heat dissipating agents, biocompatible agents and/or fibrous reinforcing materials.

The thickener comprises polycyanoacrylate, polylactic acid, polyglycolic acid, polycaprolactone, polyalkylacrylate, and polyalkylmethacrylate; stabilizer packageIncluding anionic stabilizers and free radical stabilizers, the former being metaphosphoric acid, maleic anhydride, alkylsulfonic acid, phosphorus pentoxide, iron (III) chloride, antimony oxide, 2,4, 6-trinitrophenol, mercaptans, alkylsulfonates, alkyl sulfones, alkyl sulfoxides, alkyl sulfites, sultones, sulfur dioxide and sulfur trioxide; the latter are hydroquinone, catechol and derivatives of the above compounds; initiators or accelerators include: molecules with nucleophilic function, organic or inorganic or their mixture, selected from amino, quaternary amine, hydroxyl, thiol, phosphorus-containing compound, preferably NaHCO₃，Na₂CO₃Or sodium phosphate; the colorant is selected from dyes, pigments, including PGA microfibrils, collagen microfibrils, cellulose microfibrils and olefinic microfibrils; the plasticizer comprises polyethylene glycol ester, butyl stearate, lauric acid, dioctyl glutarate, triglyceride, dioctyl adipate, triethyl phosphate, triethyl citrate, acetyl triethyl citrate and acetyl tributyl citrate; preservatives include those conventionally used but which do not initiate polymerization of the monomers and are selected from potassium sorbate, sodium benzoate, sorbic acid, chlorocresol; the heat sink comprises a liquid miscible with the monomer that evaporates during polymerization, releasing heat from the composition; the biocompatible agent comprises sodium bisulfite; the fibrous reinforcing material comprises natural or synthetic rubber to enhance the impact resistance of the composition, preferably styrene or acrylonitrile.

In a preferred embodiment, the composition may further comprise one or more biological or therapeutic agents including anti-inflammatory analgesics, sedatives, local anesthetics, non-steroidal anti-inflammatory drugs, anti-allergic drugs, anti-ulcer drugs, antibiotics, antibacterial drugs, antiviral drugs, antifungal drugs, immunosuppressive agents, naturally derived or genetically engineered proteins, polysaccharides, glycoproteins or ester proteins, oligonucleotides, polypeptide drugs, antibodies, antigens, chemotherapeutic drugs, procoagulants, hemostatic agents, and the like.

Wherein the procoagulant or hemostatic agent is prothrombin, thrombin, fibrinogen, fibrin, fibronectin, blood coagulation factor, tissue factor, collagen, gelatin, vasopressin, plasminogen activator inhibitor, platelet activator, synthetic peptide having hemostatic activity, etc.

The monomer OGA of the material provided by the invention_n-CA or the composition can be used in medicine. Such as medical adhesives or tissue engineering materials, etc. The medical adhesive can be used for wound adhesion, hemostasis, viscera and soft tissue wound closure, covering, leaking stoppage, hard tissue fixation and the like. The material monomer OGA_nthe-CA or the composition can also be used for preparing medicines for embolization of varicose veins or closed eye cornea surgery.

The invention also provides a polymer, which is obtained by crosslinking and copolymerizing a material monomer OGAN-CA or a composition under the action of anions. The anion being-OH and/or-NH₂-OH and/or-NH₂Derived from at least one of tissue, body fluid, skin and blood.

The invention is further illustrated by the following specific examples, which, however, are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.

EXAMPLE 1 Material monomer OGA_nPreparation of-CA

(ii) material monomer OGA₁-CA

The synthetic route is as follows:

the specific operation is as follows:

(1) a500 mL round-bottom flask was charged with 28g of Compound 1, 53.05g of benzyl bromide, 60mL of triethylamine, and 300mL of dichloromethane in this order, and stirred at room temperature overnight before TLC to monitor completion of the reaction. Dichloromethane and triethylamine were distilled off under reduced pressure, 300mL dichloromethane were added and redissolved, washed 2 times with 150mL saturated aqueous ammonium chloride solution, the organic layer was dried over magnesium sulfate overnight, filtered and concentrated using petroleum ether: ethyl acetate (volume ratio 5: 1) was chromatographed to give 31.8g of a colorless transparent liquid (Compound 2). The yield was 67%.

(2) 21g (compound 3), 3g DMAP and 300mL dichloromethane are added into a 500mL round-bottom flask, after the reaction solution is stirred to be clear, 16g compound 2 is added, and after stirring is continued for 10 minutes, 20g DCC is added. After stirring overnight at room temperature the reaction was monitored by TLC, filtered and concentrated, petroleum ether: ethyl acetate 5: purification on column 1 gave a colorless solid (compound 4). 80% yield.

(3) Accurately weighing 10g of compound 4, dissolving the compound 4 in 150mL of ethyl acetate, adding 10 percent by weight of palladium-carbon, reacting for about 5 hours in a hydrogen atmosphere, and reacting with dichloromethane: methanol 10: 1TLC spot plate monitor reaction. After the palladium on carbon was filtered off, the solvent was dried by spin-drying to obtain a colorless transparent solid (compound 5). Yield of 97%.

(4) Weighing 4g of compound 5, 2.7g of maleic anhydride, 10mg of phosphorus pentoxide, 5mg of hydroquinone and 50mL of dimethylbenzene, heating and refluxing at 140 ℃ for 8h, spinning off the dimethylbenzene, and repeatedly dissolving with benzene and distilling under reduced pressure for three times. 20mL of benzene was added, cooled at 4 ℃ for 2h, filtered of solid impurities, and after removal of the benzene under reduced pressure, the residue was purified by distillation with dichloromethane: diethyl ether (volume ratio 1: 4) was recrystallized at-20 deg.C, the excess maleic anhydride was filtered off while cold, and then recrystallized by redistilled dichloromethane and n-hexane at normal temperature to give 0.93g of a pale yellow oil (compound 6) in 50% yield.

(II) Material monomer OGA₂-CA

The synthetic route is as follows:

the specific operation is as follows:

(1) a250 mL round bottom flask was charged with 21.08g of Compound 7 and 22.65g of imidazole, 25mL of DMF (N, N-dimethylformamide) and finally 25.67g of t-butyldimethylchlorosilane and stirred overnight at room temperature under nitrogen. The resulting mixture was poured into 150mL of saturated aqueous sodium bicarbonate and extracted four times with 150mL of hexane. Using anhydrous MgSO₄Drying, filtering, evaporating the solvent under reduced pressure, and concentrating to obtain petroleum ether: ethyl acetate (10: 1 by volume) was chromatographed to give 24.89g of the product (Compound 8) in 80% yield.

(2) A250 mL round-bottom flask was charged with 10g of Compound 8, 100mL of distilled water was added, the mixture was stirred at 80 ℃ overnight, concentrated under reduced pressure, cooled at-20 ℃ for crystallization, and the solid was collected by filtration to give 7.5g of the product (Compound 9) in 80% yield.

(3) 7.43g of Compound 2, 8.66g of Compound 9, 7.63g of dicyclohexylcarbodiimide and 1.74g of pyridine 4-methylbenzenesulfonate are weighed out. After dissolving in 100mL of dichloromethane, the reaction was stirred at room temperature overnight. The insoluble matter was filtered, and washed with 100mL of an aqueous sodium bicarbonate solution and 100mL of an aqueous sodium chloride solution in this order. The magnesium sulfate was dried, filtered, concentrated under reduced pressure, and chromatographed to give 11.94g of the product (Compound 10) in 83% yield.

(4) 25.2g of glacial acetic acid was added to tetrahydrofuran (70mL) containing 12.09g of Compound 10, and 65.82g of tetrabutylammonium fluoride was slowly added and stirred at room temperature overnight. The product was poured into 300mL of ethyl acetate and 300mL of deionized water, and the organic layer was washed twice with 200mL of saturated sodium bicarbonate, 5% citric acid, and saturated aqueous sodium chloride solution in this order, dried over magnesium sulfate, filtered, distilled under reduced pressure, and chromatographed to give 7.7g of the product (compound 11) in 80% yield.

(5) 21g of anthracenecyanoacrylic acid (Compound 3), DMAP3g and 300mL of DCM were added to a 500mL round-bottomed flask, and after stirring until the reaction solution became clear, 25g of Compound 11 was added, and after stirring for 10 minutes, 20g of DCC (dicyclohexylcarbodiimide) was added. Stir at rt overnight, TLC monitored for reaction completion, filtered and concentrated, petroleum ether: ethyl acetate (5: 1 by volume) was chromatographed to give 35.69g of a white solid (Compound 12) in 70% yield.

(6) 10g of compound 12 and 1g of palladium-carbon were weighed, and ethyl acetate was used as a solvent, and nitrogen was introduced into a reaction flask, and the reaction was stirred at room temperature overnight. The palladium on carbon was filtered using filter paper, ethyl acetate was evaporated under reduced pressure, and the mixture was purified as dichloromethane: the product was isolated and purified by column chromatography using methanol (10: 1 by volume) as eluent to give 7.86g of a white solid (Compound 13) in 90% yield.

(7) Weighing 4g of compound 13, 2.3g of maleic anhydride, 10mg of phosphorus pentoxide, 5mg of hydroquinone and 50mL of dimethylbenzene, heating and refluxing at 140 ℃ for 8h, spinning off the dimethylbenzene, and repeatedly dissolving with benzene and distilling under reduced pressure for three times. 20mL of benzene was added, cooled at 4 ℃ for 2h, filtered of solid impurities, and after removal of the benzene under reduced pressure, the residue was purified by distillation with dichloromethane: diethyl ether (volume ratio 1: 4) was recrystallized at-20 deg.C, the excess maleic anhydride was filtered off while cold, and then recrystallized by redistilled dichloromethane and n-hexane at normal temperature to give 1.1g of a pale yellow oil (compound 14) in 50% yield.

(III) Material monomer OGA₃-CA

The synthetic route is as follows:

the specific operation is as follows:

(1) accurately weighing 10g of compound 10, dissolving in 150mL of ethyl acetate, adding 10 percent by weight of palladium-carbon, reacting for about 5 hours in a hydrogen atmosphere, and reacting with dichloromethane: methanol 10: 1TLC spot plate monitor reaction. After the palladium on carbon was filtered off, the solvent was evaporated by rotary evaporation to obtain a colorless transparent solid (compound 15). Yield of 97%.

(2) 10g of Compound 15 and 2g of DMAP were weighed out accurately and dissolved in 250mL of dichloromethane, and after stirring and reacting for 10 minutes at normal temperature, 23.73 g of Compound and 3.96g of DCC were added. The reaction was stirred at room temperature overnight, petroleum ether: ethyl acetate 5: the reaction was monitored on a 1-point plate. Insoluble white solid was filtered and concentrated, petroleum ether: ethyl acetate 5: purification on column 1 gave a colorless solid (compound 16). 75% yield.

(3) 25.2g of glacial acetic acid were added to tetrahydrofuran (70mL) containing 12.09g of Compound 16, and 65.82g of tetrabutylammonium fluoride was slowly added and stirred at room temperature overnight. The product was poured into 300mL of ethyl acetate and 300mL of deionized water, and the organic layer was washed twice with 200mL of saturated sodium bicarbonate, 5% citric acid and saturated aqueous sodium chloride solution in this order, dried over magnesium sulfate, filtered, distilled under reduced pressure, and chromatographed to give 7.7g of the product (compound 17) in 80% yield.

(4) 7.43g of Compound 3, 8.66g of Compound 17, 7.63g of dicyclohexylcarbodiimide and 1.74g of pyridine 4-methylbenzenesulfonate are weighed out. After dissolving in 100mL of dichloromethane, the reaction was stirred at room temperature overnight. The insoluble matter was filtered, and washed with 100mL of an aqueous sodium bicarbonate solution and 100mL of an aqueous sodium chloride solution in this order. The magnesium sulfate was dried, filtered, concentrated under reduced pressure, and chromatographed to give 11.94g of the product (Compound 18) in 83% yield.

(5) 10g of compound 18 and 1g of palladium-carbon were weighed, and ethyl acetate was used as a solvent, and a reaction flask was filled with nitrogen, and stirred at room temperature for reaction overnight. The palladium on carbon was filtered using filter paper, ethyl acetate was evaporated under reduced pressure, and the mixture was purified as dichloromethane: the product was isolated and purified by column chromatography using methanol (10: 1 by volume) as eluent to give 7.86g of a white solid (Compound 19) in 90% yield.

(6) Weighing 4g of compound 19, 2g of maleic anhydride, 10mg of phosphorus pentoxide, 5mg of hydroquinone and 50mL of xylene, heating and refluxing at 140 ℃ for 8h, spinning off the xylene, and repeatedly dissolving with benzene and distilling under reduced pressure for three times. 20mL of benzene was added, cooled at 4 ℃ for 2h, filtered of solid impurities, and after removal of the benzene under reduced pressure, the residue was purified by distillation with dichloromethane: diethyl ether (volume ratio 1: 4) was recrystallized at-20 deg.C, the excess maleic anhydride was filtered off while cold, and then recrystallized by redistilled dichloromethane and n-hexane at normal temperature to give 1.2g of a pale yellow oil (compound 20) in 50% yield.

(IV) Material monomer OGA₄-CA to OGA₁₀-CA

Referring to the above synthetic routes, monomers of materials with n ═ 4, 5, 6, 7, 8, 9, and 10 were synthesized, respectively.

Example 2 polymeric phase Change and Nuclear magnetic characterization of materials

With material monomer OGA₂CA is for example a liquid before use, which polymerizes under anionic initiation conditions (e.g.initiated by water vapour when placed in a humid environment), from a liquid state to a solid state. The change of the characteristic peaks of nuclear magnetism before and after the monomer polymerization is further tested through nuclear magnetism, and the result is shown in figure 1, the characteristic peaks of hydrogen atoms on the olefinic double bond of the monomer structure before the polymerization (chemical shifts are 6.7ppm and 7.1ppm), and a wide unshaped peak is formed after the polymerization is shifted to the vicinity of 2.6ppm, which indicates that the olefinic double bond in the monomer is completely converted into a polycarbonic chain.

Detecting monomer OGA of material by nuclear magnetic characterization_nPolymerization and curing of the-CA's all take place.

The above experiments show that this material system has the typical characteristics of polymerizable curing based on cyanoacrylate functionality in the monomer structure.

Example 3 Performance testing

Monomer curing time

OGA_nAfter the-CA series monomer is contacted with the pigskin tissue, the liquid state can be completely changed into the solid state within 5 minutes, and nuclear magnetic analysis proves that the olefinic double bond in the monomer is completely polymerized. The polymerization rate of this system is greatly reduced compared to conventional n-butyl cyanoacrylate adhesives (within 30 s), probably because the steric hindrance of the OGA chain in the monomer molecule and the inhibition of the terminal carboxyl group affect the thermodynamic collision efficiency of the olefinic double bonds, resulting in an extension of the through-cure time.

Shear tensile strength

Shear tensile strength is a common indicator of adhesive bonding ability. The national pharmaceutical industry standard (YY/T0729-2009) is taken as the experimental guidance. Preparing long-strip pigskin (2.5cm is multiplied by 7.5cm), sucking 20 mu L of monomer to drip on the pigskin 2.5cm is multiplied by 1cm²The areas with the same size are quickly and uniformly coated, another pigskin with the same size is quickly lapped on the pigskin, the pressure in the vertical direction of 10N is 10min, a FGS-500TW-SL tensile testing machine is used for testing the maximum tensile force when the bonding area of the two pigskins is broken at a constant speed of 20mm/min, and six groups of parallel tests are set for each sample. The results are given in the table below.

Polymer molecular weight determination

The physicochemical properties of polymers are closely related to their molecular weight. The molecular weight of the polymer was analyzed by molecular gel exclusion chromatography, and the results are shown in the following table, wherein the polymer (oligomer) formed by polymerization and solidification of each monomer has a weight average molecular weight of 5000-7000 dalton (Da) and a more concentrated molecular weight distribution (PDI ═ 1.15-1.35).

Example 4 Biosafety

Cytotoxicity

Part 5 of the biological evaluation of medical devices according to GB/T16886.5-2017: the in vitro cytotoxicity test method adopts sterile normal saline injection and sterile normal saline injection with the volume fraction of 2 percent DMSO as leaching liquor to carry out MTT method test. Adding the product into 6-well plate under aseptic condition, 10 μ L per well, and spreading uniformly in a medium containing 5% CO₂After 2 days of curing in a 37 ℃ incubator. According to 6cm²The leaching solution is added in the ratio of/mL, and leached for 72 hours at 37 ℃ under the condition of shaking at 100 rpm. The cells were diluted to 25%, 12.5%, 6.25%, and 3.125% with serum-containing MEM medium before being contacted, while blank, negative, and positive controls were set. NCTCL929 cells in a logarithmic growth phase are selected for the test, the appearance of the cells is observed after the sample is contacted with the cells for 24 hours, and the survival rate of the cells is determined by an MTT method.

The experimental results are as follows: observation of cell morphology the non-polar leach liquor and the polar leach liquor of each concentration of the product have discrete particles in the cytoplasm of cultured cells, and no cell is dissolved and no cell proliferation is reduced. The cell survival rate of 25% of the nonpolar leaching liquor of the monomer is 95% and the cell survival rate of 25% of the polar leaching liquor is 93% by MTT method detection. The material has no potential cytotoxicity reaction of 25% non-polar leaching liquor and 25% polar leaching liquor.

Sensitization

Part 10 of the biological evaluation of medical devices according to GB/T16886.10-2005: and in the stimulation and delayed hypersensitivity tests, 0.9 percent sodium chloride injection and fresh vegetable oil are selected as leaching liquor, 0.1mL is injected into each injection point in an intradermal way, and the skin condition of the animal at the excitation part is observed and recorded after 24 hours and 48 hours of excitation.

Experimental materials and instruments: guinea pig, Freund's adjuvant (SIGMA F5881-10ml CAS9007-81-2), 0.9% sodium chloride injection, cotton seed oil, syringe, the product.

The experimental results are as follows: the skin of the stimulated part of the animal in the test group is not stimulated positively, the positive stimulation incidence rate is 0 percent, and the material is proved to have no skin sensitization reaction.

Intradermal reaction

Part 10 of the biological evaluation of medical devices according to GB/T16886.10-2005: stimulation and delayed hypersensitivity testing

Experimental materials and instruments: healthy rabbits, syringes, 0.9% sodium chloride injection, cottonseed oil and the product.

The experimental results are as follows: after the leaching liquor of the product is injected, the difference of the average scores of the product group and the control group is less than 1.0, and the product has no skin irritation.

Skin irritation

Part 10 of the biological evaluation of medical devices according to GB/T16886.10-2005: stimulation and delayed type hypersensitivity tests, the skin condition of the application site was observed at 1, 24, 48 and 72 hours after 0.1mL of the product was applied to the skin of rabbits at home and blocked for 4 hours.

Experimental materials and instruments: rabbit, the product, depilatory, pipettor, medical gauze.

The experimental results are as follows: no erythema and/or edema symptoms appear on the skin of the experimental area, and the product has no irritant effect on the skin of the rabbits.

Acute systemic toxicity

(1) The experimental method comprises the following steps: referring to GB/T-14233.2-2005, mice were gavaged with 10g/kg of the material monomer polymer.

The experimental results are as follows: after the mice are subjected to gastric lavage, no toxic reaction occurs, the mice are killed and examined for diseases after 4 days, and no abnormality is observed by naked eyes.

(2) The experimental method comprises the following steps: the polymer colloid leaching liquor (leaching ratio is 1.25 cm)²mL, the leaching medium is sterile physiological saline) are respectively injected into tail vein and abdominal cavity, and the dosage is 50 mL/kg.

Experimental materials and instruments: 2% of methyl cellulose solution, the product, Kunming mice and an electronic balance.

The experimental results are as follows: no significant toxicity was observed, mice did not show special toxicant symptoms, no death, and normal weight gain, similar to normal controls.

The experiments indicate that no acute toxicity of mice is observed under the experimental conditions of the product.

Hemolysis of blood

The experimental method is carried out according to GB/T14233.2-2005, GB/T16886.12-2017. Healthy adult rabbits with the body weight of more than 2.0kg are selected, blood is collected from the heart on the test day, and diluted anticoagulated rabbit blood is prepared after anticoagulation. The test sample contained 5% CO₂Curing the mixture in a constant-temperature incubator at 37 ℃ for 2d, adding 0.9% sodium chloride injection, leaching for 72h at 37 ℃, and respectively collecting 10mL of leaching liquor to test sample tubes; adding 10mL of 0.9% sodium chloride injection into each tube of the negative control group; the positive control group added 10mL of purified water per tube. Each set operated 3 tubes in parallel. And (3) after the temperature is kept for 30min, 0.2mL of diluted anticoagulated rabbit blood is added into each test tube, and the temperature is kept for 60min after the mixture is uniformly mixed. After the liquid in the tube was poured out, it was centrifuged, and absorbance was measured at a wavelength of 545nm with an ultraviolet spectrophotometer, and the hemolysis ratio (%) of the test sample was calculated.

Experimental materials and instruments: adult rabbit, anticoagulant, 0.9% sodium chloride injection, the product, a constant temperature water bath kettle and an ultraviolet spectrophotometer.

The experimental results are as follows: the material monomer has the hemolysis rate of 0.1% under the above experimental conditions, has no obvious hemolysis risk, and meets the GB/T14233.2-2005 standard.

The polymerization degree of the OGAN-CA system is far lower than that of the traditional CA material (for example, the weight average molecular weight of n-butyl cyanoacrylate after polymerization under physiological conditions is hundreds of thousands or even millions), and the polymerization degree is probably related to that the steric hindrance effect of the OGA chain greatly reduces the probability of the reaction of the proximity of the ethylenic bond functional groups, and the reason that the bonding strength of the system is lower than that of the n-butyl cyanoacrylate (about 300-500 kPa) is probably also the main reason. From a degradation point of view, this relatively low molecular weight is more favorable for metabolism: the polylactic acid has good biodegradability, and the side chain oligomeric lactic acid is degraded to continuously increase water solubility, and the lower molecular weight of the polymer is beneficial to direct absorption and metabolism of degradation products, so that the system has potential biodegradable and metabolizable characteristics.

The invention synthesizes material monomers containing cyanoacrylate functional groups and oligoglycolic acid segments (OGA) by a chemical synthesis mode. The monomer has small molecular weight and is liquid at normal temperature, and can be polymerized under the condition of anion due to the cyanoacrylate functional group. The obtained polymer contains a large amount of OGA side chains, so that the polymer has good biocompatibility and softness, and more importantly, the OGA contains a plurality of ester bonds easy to hydrolyze, so that the polymer can be endowed with good degradation performance. The physical and chemical properties of the polymer can be further regulated and controlled by regulating the number of OGA repeating units, so that different practical application requirements are met.

While particular embodiments of the present invention have been illustrated and described, it would be obvious that various other changes and modifications can be made without departing from the spirit and scope of the invention. It is therefore intended to cover in the appended claims all such changes and modifications that are within the scope of this invention.