CN107573451B - A kind of beta-lactam antibiotic graft polymers and its application - Google Patents
A kind of beta-lactam antibiotic graft polymers and its application Download PDFInfo
- Publication number
- CN107573451B CN107573451B CN201710596048.7A CN201710596048A CN107573451B CN 107573451 B CN107573451 B CN 107573451B CN 201710596048 A CN201710596048 A CN 201710596048A CN 107573451 B CN107573451 B CN 107573451B
- Authority
- CN
- China
- Prior art keywords
- beta
- lactam antibiotic
- graft polymers
- sample
- column
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000003782 beta lactam antibiotic agent Substances 0.000 title claims abstract description 48
- 229920000578 graft copolymer Polymers 0.000 title claims abstract description 48
- 239000002132 β-lactam antibiotic Substances 0.000 title claims abstract description 48
- 229940124586 β-lactam antibiotics Drugs 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 claims abstract description 20
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000003814 drug Substances 0.000 claims abstract description 15
- 125000004029 hydroxymethyl group Chemical group [H]OC([H])([H])* 0.000 claims abstract description 15
- 238000001514 detection method Methods 0.000 claims abstract description 14
- 229940079593 drug Drugs 0.000 claims abstract description 11
- 229920005990 polystyrene resin Polymers 0.000 claims abstract description 11
- 239000007788 liquid Substances 0.000 claims abstract description 10
- XJRPTMORGOIMMI-UHFFFAOYSA-N ethyl 2-amino-4-(trifluoromethyl)-1,3-thiazole-5-carboxylate Chemical compound CCOC(=O)C=1SC(N)=NC=1C(F)(F)F XJRPTMORGOIMMI-UHFFFAOYSA-N 0.000 claims abstract description 9
- 241000894006 Bacteria Species 0.000 claims abstract description 8
- 238000007142 ring opening reaction Methods 0.000 claims abstract description 8
- 239000003054 catalyst Substances 0.000 claims abstract description 7
- 239000006166 lysate Substances 0.000 claims abstract description 7
- 229920000642 polymer Polymers 0.000 claims abstract description 5
- 238000012544 monitoring process Methods 0.000 claims abstract description 3
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 16
- 239000000243 solution Substances 0.000 claims description 13
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 12
- 239000003960 organic solvent Substances 0.000 claims description 12
- 238000006243 chemical reaction Methods 0.000 claims description 11
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 10
- 239000012065 filter cake Substances 0.000 claims description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 8
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 8
- 239000000872 buffer Substances 0.000 claims description 7
- 238000002360 preparation method Methods 0.000 claims description 7
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 6
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 6
- 239000000047 product Substances 0.000 claims description 6
- 239000007983 Tris buffer Substances 0.000 claims description 5
- 230000001580 bacterial effect Effects 0.000 claims description 5
- 102000006635 beta-lactamase Human genes 0.000 claims description 5
- 238000004321 preservation Methods 0.000 claims description 5
- 230000003068 static effect Effects 0.000 claims description 5
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 5
- SCYULBFZEHDVBN-UHFFFAOYSA-N 1,1-Dichloroethane Chemical class CC(Cl)Cl SCYULBFZEHDVBN-UHFFFAOYSA-N 0.000 claims description 4
- 229930186147 Cephalosporin Chemical class 0.000 claims description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 4
- 108090000204 Dipeptidase 1 Proteins 0.000 claims description 4
- 229940124587 cephalosporin Drugs 0.000 claims description 4
- 150000001780 cephalosporins Chemical class 0.000 claims description 4
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 claims description 4
- 229930182555 Penicillin Natural products 0.000 claims description 3
- 235000009508 confectionery Nutrition 0.000 claims description 3
- 230000035945 sensitivity Effects 0.000 claims description 3
- 229910052708 sodium Inorganic materials 0.000 claims description 3
- 239000011734 sodium Substances 0.000 claims description 3
- WZPBZJONDBGPKJ-VEHQQRBSSA-N aztreonam Chemical class O=C1N(S([O-])(=O)=O)[C@@H](C)[C@@H]1NC(=O)C(=N/OC(C)(C)C(O)=O)\C1=CSC([NH3+])=N1 WZPBZJONDBGPKJ-VEHQQRBSSA-N 0.000 claims description 2
- 229940041011 carbapenems Drugs 0.000 claims description 2
- 238000006555 catalytic reaction Methods 0.000 claims description 2
- 150000002960 penicillins Chemical class 0.000 claims description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims 3
- GSNUFIFRDBKVIE-UHFFFAOYSA-N DMF Natural products CC1=CC=C(C)O1 GSNUFIFRDBKVIE-UHFFFAOYSA-N 0.000 claims 1
- 206010059866 Drug resistance Diseases 0.000 abstract description 10
- 238000004458 analytical method Methods 0.000 abstract description 7
- 108090000623 proteins and genes Proteins 0.000 abstract description 4
- 230000001413 cellular effect Effects 0.000 abstract description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 abstract description 3
- 102000004169 proteins and genes Human genes 0.000 abstract description 3
- 230000000694 effects Effects 0.000 abstract description 2
- 238000005406 washing Methods 0.000 abstract 1
- 102000020235 metallo-beta-lactamase Human genes 0.000 description 10
- 108060004734 metallo-beta-lactamase Proteins 0.000 description 10
- HVFLCNVBZFFHBT-ZKDACBOMSA-N cefepime Chemical compound S([C@@H]1[C@@H](C(N1C=1C([O-])=O)=O)NC(=O)\C(=N/OC)C=2N=C(N)SC=2)CC=1C[N+]1(C)CCCC1 HVFLCNVBZFFHBT-ZKDACBOMSA-N 0.000 description 8
- 229960002100 cefepime Drugs 0.000 description 8
- 150000003952 β-lactams Chemical class 0.000 description 7
- 230000003115 biocidal effect Effects 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- KAESVJOAVNADME-UHFFFAOYSA-N 1H-pyrrole Natural products C=1C=CNC=1 KAESVJOAVNADME-UHFFFAOYSA-N 0.000 description 3
- KMIPKYQIOVAHOP-YLGJWRNMSA-N cefditoren Chemical compound S([C@@H]1[C@@H](C(N1C=1C(O)=O)=O)NC(=O)\C(=N/OC)C=2N=C(N)SC=2)CC=1\C=C/C=1SC=NC=1C KMIPKYQIOVAHOP-YLGJWRNMSA-N 0.000 description 3
- 229960004069 cefditoren Drugs 0.000 description 3
- -1 pyrrole oxime Chemical class 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 101710116957 D-alanyl-D-alanine carboxypeptidase Proteins 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- HSRJKNPTNIJEKV-UHFFFAOYSA-N Guaifenesin Chemical compound COC1=CC=CC=C1OCC(O)CO HSRJKNPTNIJEKV-UHFFFAOYSA-N 0.000 description 2
- 239000004793 Polystyrene Substances 0.000 description 2
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 229920002223 polystyrene Polymers 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 235000011121 sodium hydroxide Nutrition 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 108020004256 Beta-lactamase Proteins 0.000 description 1
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000002814 agar dilution Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000012742 biochemical analysis Methods 0.000 description 1
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- 229940036735 ceftaroline Drugs 0.000 description 1
- ZCCUWMICIWSJIX-NQJJCJBVSA-N ceftaroline fosamil Chemical compound S([C@@H]1[C@@H](C(N1C=1C([O-])=O)=O)NC(=O)\C(=N/OCC)C=2N=C(NP(O)(O)=O)SN=2)CC=1SC(SC=1)=NC=1C1=CC=[N+](C)C=C1 ZCCUWMICIWSJIX-NQJJCJBVSA-N 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 229920006389 polyphenyl polymer Polymers 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 108700026220 vif Genes Proteins 0.000 description 1
Landscapes
- Cephalosporin Compounds (AREA)
Abstract
The present invention provides a kind of beta-lactam antibiotic graft polymers and its applications, comprising the following steps: under the effect of the catalyst, hydroxymethyl polystyrene resin and bisglycidyl ether are condensed；The obtained amino on product and 4 side chains of beta-lactam antibiotic carries out ring-opening reaction, by washing repeatedly, obtains the graft polymers of the drug resistance of detection beta-lactam antibiotic；Polymer is filled into column, utilizes the appearance time of sample in UV detector monitoring efflux；Bromocresol purple detection is carried out to packed column, to be quickly obtained sample to the sensitive information of drug.This method is applicable not only to protein sample, is also applied for the drug resistance of bacterium living body and cellular lysate liquid detection beta-lactam antibiotic, therefore can be applied to clinical labororatory and carry out quick resistance analysis, and doctor is instructed to determine clinical application scheme as early as possible.
Description
Technical field
The present invention is a kind of beta-lactam antibiotic graft polymers and its application.Belong to biochemical analysis field.
Background technique
The change of penicillin binding protein (PBP) often results in the clinically important drug-resistant phenotype of the following two kinds；With beta-lactam class
The affinity of antibiotic lowers, so as to cause bacterium to beta-lactam antibiotic drug resistance；Acquired resistance is by plasmid-mediated
, after bacterium obtains drug resistant gene, a large amount of beta-lactamases (rather than PBPs) is generated, inactivates penicillinase-fast penicillin slowly, table
Reveal drug resistance, mostly critical drug resistance；Traditional antibiotics sensitivity detection beta-lactam antibiotic method is agar dilution
Method, i.e. minimal inhibitory concentration MIC detect beta-lactam antibiotic method.The advantages of this method is quantitative, accurate, major defect
It is that complicated for operation time-consuming.In recent years, the Drug Resistance Detection beta-lactam for being passivated enzyme gene or biochemical activity based on antibiotic resists
Raw element method is becoming increasingly popular, and the advantages of this method is the table that can determine specific beta-lactamase from molecular level quickly
It reaches, the disadvantage is that its primer is only used for detection beta-lactam antibiotic known, it is impossible to be used in new lactamase base
The detection beta-lactam antibiotic of cause and mutated gene.Directly detection beta-lactam antibiotic bacterial lactamase is active
Experimental method also has been reported that, such as Bai Nawu, Pirelli et al. hydrolyze β-using ultraviolet spectrophotometry analysis bacterial lysate
Beta-lactam antibiotics level is to provide its drug resistance information, but this method heavy workload, and there is still a need for several hours to obtain
Test result out.
Summary of the invention
The object of the present invention is to provide a kind of beta-lactam antibiotic graft polymers and its applications, utilize hydroxymethyl
Polystyrene resin is grafted after bisglycidyl ether and carries out ring-opening reaction with the amino on 4 side chains of beta-lactam antibiotic,
Obtain detection beta-lactam antibiotic cephalosporins medicine drug resistance graft polymers；Polymer is filled into column, is flowed out by monitoring
The UV absorption variation of liquid is to be quickly obtained sample to the sensitive information of drug.It the advantage is that, supervised using UV detector
The appearance time of metallo-β-lactamase in efflux is controlled, and bromocresol purple detection is carried out to packed column, to be quickly obtained
Sensitive information of the sample to drug.To it is medicaments insensitive, lowering with the affinity of beta-lactam antibiotic, make beta-lactam
The sample of ring degradation can obtain fine analysis result；This method is applicable not only to protein sample, is also applied for bacterium living body
And cellular lysate liquid detects beta-lactam antibiotic, therefore can be applied to clinical labororatory and carry out quick resistance analysis.
The object of the present invention is achieved like this, a kind of beta-lactam antibiotic graft polymers and its application, this is poly-
Close the structural formula of object are as follows:
Wherein, R is beta-lactam antibiotic；
The preparation method of the beta-lactam antibiotic graft polymers the following steps are included:
1), in organic solvent A, under the catalysis of a small amount of catalyst, hydroxymethyl polystyrene resin and double shrinks are sweet
Oily ether condensation, reaction solution filtering, filter cake are sufficiently washed with organic solvent A, finally obtain graft polymers；
2), disperse organic solvent B for the graft polymers that the product of step 1) obtains, by beta-lactam antibiotic plus
Enter and carry out ring-opening reaction in reactor, end of reaction, reaction solution filters, and filter cake is sufficiently washed with organic solvent B, finally obtains inspection
Survey beta-lactam antibiotic graft polymers.
Catalyst used in step 1) are as follows: sodium hydroxide, SDS or 16- sodium alkyl sulfate；Hydroxymethyl polystyrene tree
The molar ratio of hydroxyl and bisglycidyl ether is 1:2-1:10 in rouge；Organic solvent A are as follows: dichloroethanes, ethyl acetate, tetrahydro furan
It mutters, DMF or MSO；
Using hydroxyl in hydroxymethyl polystyrene resin as standard, the input amount of the step 1) catalyst is 0.5%
5.5%.
Step 2) the organic solvent B are as follows: methylene chloride, chloroform, carbon tetrachloride, tetrahydrofuran or DMF；Hydroxymethyl is poly-
The molar ratio of hydroxyl and beta-lactam antibiotic is 1:2-1:8 in styrene resin；The beta-lactam antibiotic are as follows:
Penicillins, cephalosporins, Carbapenems or monobactam class.
A method of sample is quickly obtained to the sensitive information of drug using beta-lactam antibiotic graft polymers,
The following steps are included: 1., by above-mentioned steps 2) obtained beta-lactam antibiotic graft polymers is suspended in PBS, uses
Wet method dress post, every pillar are packed into lmL gel, after packed column installs, continued column with sterile buffer, will reveal to column top
Out when liquid level, stopped column, static preservation；2., sample passed through into the obtained packed column of step 3) with certain flow rate, utilization is ultraviolet
The appearance time of metallo-β-lactamase in detector monitors efflux；After sample flows completely out, bromine first is carried out to packed column
Phenol violet detection, to be quickly obtained sample to the sensitive information of drug.Wherein, 1. the PBS is 50mM Tris, pH to step
7.0, the NaN that sterile buffer is 0.01%3- PBS, volume are five times of column interpolymer volume；The step 2. sample
Are as follows: beta-lactamase, bacterial lysate or the clinical bacteria of purifying, the flow velocity of sample are 0.1ml/min.
The beneficial effects of the present invention are:
The present invention provides a kind of beta-lactam antibiotic graft polymers and its applications, utilize hydroxymethyl polyphenyl second
Olefine resin is grafted after bisglycidyl ether and carries out ring-opening reaction with the amino on 4 side chains of beta-lactam antibiotic, is examined
Survey beta-lactam antibiotic cephalosporins medicine drug resistance graft polymers；Polymer is filled into column, is monitored using UV detector
The appearance time of metallo-β-lactamase in efflux, and bromocresol purple detection is carried out to packed column, to be quickly obtained sample
Sensitive information of the product to drug.It the advantage is that, it is that medicaments insensitive, with beta-lactam class antibiotic affinity lowers, make
The sample of beta-lactam nucleus degradation can obtain fine analysis result；This method is applicable not only to protein sample, is also applied for
The analysis of the drug resistance of live bacteria and cellular lysate liquid, therefore can be applied to clinical labororatory and carry out quick resistance analysis.
Detailed description of the invention
Fig. 1 is the preparation route figure of beta-lactam antibiotic graft polymers of the present invention.
Fig. 2 metallo-β-lactamase compares in two kinds of packed columns in the delivery time.It is (red: blank packed column, packing material
For the hydroxymethyl poly styrene polymer of non-grafted antibiotic；Blue: beta-lactam antibiotic graft polymers)
Fig. 3 determines the beta-lactam nucleus hydrolyzed in packed column by metallo-β-lactamase using bromocresol purple method.(left side is
Beta-lactam antibiotic graft polymers packed column without metallo-β-lactamase elution；Right side is through Metallo-β-lactamases
The beta-lactam antibiotic graft polymers packed column of enzyme elution)
Specific embodiment
The invention patent is described in more detail with reference to the accompanying drawings and detailed description:
The preparation method of beta-lactam antibiotic graft polymers of the present invention, does furtherly in conjunction with the embodiments
It is bright, but these embodiments are not construed as the limitation as the scope of the present invention.
Specific embodiment is as follows:
Embodiment 1
The preparation of Cefepime graft polymers
Step 1: 100ml dichloroethanes being added in there-necked flask, by 10g hydroxymethyl polystyrene resin and the bis- contractings of 20g
Water glycerin ether is added in reactor and is uniformly mixed, and 40% sodium hydrate aqueous solution 1ml is slowly added dropwise under room temperature, reacts 5 hours
Afterwards, it filters, filter cake is sufficiently washed with 200ml dichloroethanes, obtains graft polymers；
Step 2: dispersing the graft polymers that the product of step 1) obtains in 100ml tetrahydrofuran solution, by 20g head
Spore pyrrole oxime, which is added in reactor, carries out ring-opening reaction, reacts 8 hours, and reaction solution filtering, filter cake is sufficiently washed with 200ml tetrahydrofuran
It washs, finally obtains Cefepime graft polymers；
Step 3: the Cefepime graft polymers that step 2) obtains is suspended in 50mM Tris, in the PBS of pH 7.0,
Using wet method dress post, every pillar is packed into l ml gel, after pillar installs, with 5ml, 0.01%NaN3- PBS sterile buffer after
Continue column, when column top will expose liquid level, stops column, static preservation.
Embodiment 2
The preparation of ceftaroline graft polymers
Step 1: 100ml tetrahydrofuran being added in there-necked flask, by 10g hydroxymethyl polystyrene resin and the bis- contractings of 20g
Water glycerin ether is added in reactor and is uniformly mixed, and 0.3g SDS is slowly added under room temperature, after reaction 5 hours, filtering, filter cake is used
200ml tetrahydrofuran sufficiently washs, and obtains graft polymers；
Step 2: dispersing the graft polymers that the product of step 1) obtains in 100ml DMF solution, by 20g cephalo pyrrole
Oxime, which is added in reactor, carries out ring-opening reaction, reacts 8 hours, and reaction solution filtering, filter cake is sufficiently washed with 200mlDMF, final
To Cefepime graft polymers；
Step 3: the Cefepime graft polymers that step 2) obtains is suspended in 50mM Tris, in the PBS of pH 7.0,
Using wet method dress post, every pillar is packed into l ml gel, after pillar installs, with 5ml, 0.01%NaN3- PBS sterile buffer after
Continue column, when column top will expose liquid level, stops column, static preservation.
Embodiment 3
The preparation of cefditoren graft polymers
Step 1: 100ml DMF is added in there-necked flask, 10g hydroxymethyl polystyrene resin and the bis- shrinks of 20g is sweet
Oily ether is added in reactor and is uniformly mixed, and 0.4g 16- sodium alkyl sulfate is slowly added under room temperature and is filtered after reaction 5 hours, filter
Cake is sufficiently washed with 200ml DMF, obtains graft polymers；
Step 2: dispersing the graft polymers that the product of step 1) obtains in 100ml dichloromethane solution, by 20g head
Spore pyrrole oxime, which is added in reactor, carries out ring-opening reaction, reacts 8 hours, and reaction solution filtering, filter cake is sufficiently washed with 200ml methylene chloride
It washs, finally obtains cefditoren graft polymers；
Step 3: the cefditoren graft polymers that step 2) obtains is suspended in 50mM Tris, the PBS of pH 7.0
In.Using wet method dress post, every pillar is packed into l ml gel, after pillar installs, with 5ml, 0.01%NaN3- PBS sterile buffer
Liquid continued column, when column top will expose liquid level, stopped column, static preservation.
Embodiment 4
The application of Cefepime graft polymers
Metallo-β-lactamase solution is passed through into the packed column that step 3) obtains with certain flow rate, utilizes connection packed column
UV detector detects the appearance time of the metallo-β-lactamase in efflux (see Fig. 2)；After sample flows completely out, use
0.04% bromocresol purple solution elutes packed column, until color is full of packed column.Bromocresol purple testing result is shown in figure
3.The result shows that Cefepime graft polymers packed column has apparent hesitation to metallo-β-lactamase；Bromocresol purple method
Testing result shows that the beta-lactam on Cefepime graft polymers packed column is broken.
Claims (5)
1. a kind of beta-lactam antibiotic graft polymers, it is characterised in that: the structural formula of the polymer are as follows:
Wherein, R is beta-lactam antibiotic；
The preparation method of the beta-lactam antibiotic graft polymers the following steps are included:
1), in organic solvent A, under the catalysis of catalyst, hydroxymethyl polystyrene resin and bisglycidyl ether are condensed,
Reaction solution filtering, filter cake are sufficiently washed with organic solvent A, finally obtain graft polymers；With hydroxymethyl polystyrene resin
Middle hydroxyl is standard, and the input amount of the catalyst is 0.5%~5.5%；
2) organic solvent B, is dispersed by the graft polymers that the product of step 1) obtains, beta-lactam antibiotic is added anti-
It answers and carries out ring-opening reaction in device, end of reaction, reaction solution filters, and filter cake is sufficiently washed with organic solvent B, finally obtains detection β-
Lactam antibiotics graft polymers.
2. a kind of beta-lactam antibiotic graft polymers according to claim 1, it is characterised in that: institute in step 1)
Catalyst are as follows: sodium hydroxide, SDS or 16- sodium alkyl sulfate；Hydroxyl and double shrinks are sweet in hydroxymethyl polystyrene resin
The molar ratio of oily ether is 1:2-1:10；Organic solvent A are as follows: dichloroethanes, ethyl acetate, tetrahydrofuran, DMF or DMSO.
3. a kind of beta-lactam antibiotic graft polymers according to claim 1, it is characterised in that: step 2) is described
Organic solvent B are as follows: methylene chloride, chloroform, carbon tetrachloride, tetrahydrofuran or DMF；In hydroxymethyl polystyrene resin hydroxyl with
The molar ratio of beta-lactam antibiotic is 1:2-1:8；The beta-lactam antibiotic are as follows: penicillins, cephalosporin
Class, Carbapenems or monobactam class.
4. a kind of beta-lactam antibiotic graft polymers is being quickly obtained sample to the sensitivity of drug as described in claim 1
Application in information, the sample are beta-lactamase, bacterial lysate or the clinical bacteria of purifying.
5. a kind of be quickly obtained sample to the sensitivity of drug using beta-lactam antibiotic graft polymers described in claim 1
The method of information, it is characterised in that: the following steps are included: the beta-lactam antibiotic 1., by step 2) obtained is graft-polymerized
Object is suspended in PBS, and using wet method dress post, every pillar is packed into lmL gel, after packed column installs, is continued with sterile buffer
Column is crossed, when column top will expose liquid level, stopped column, static preservation；2., by sample with certain flow rate by step 3)
The packed column arrived using the appearance time of sample in UV detector monitoring efflux, and carries out bromocresol purple to packed column
Detection, to be quickly obtained sample to the sensitive information of drug；Wherein, 1. the PBS is 50mM Tris, pH 7.0 to step,
The NaN that sterile buffer is 0.01%3- PBS, volume are five times of column interpolymer volume；The step 2. sample are as follows: pure
Beta-lactamase, bacterial lysate or the clinical bacteria of change；The bromocresol purple solution concentration is 0.04%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710596048.7A CN107573451B (en) | 2017-07-20 | 2017-07-20 | A kind of beta-lactam antibiotic graft polymers and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710596048.7A CN107573451B (en) | 2017-07-20 | 2017-07-20 | A kind of beta-lactam antibiotic graft polymers and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107573451A CN107573451A (en) | 2018-01-12 |
| CN107573451B true CN107573451B (en) | 2019-11-08 |
Family
ID=61050023
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710596048.7A Active CN107573451B (en) | 2017-07-20 | 2017-07-20 | A kind of beta-lactam antibiotic graft polymers and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107573451B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4209426A (en) * | 1979-05-29 | 1980-06-24 | American Home Products Corporation | Polypeptides related to somatostatin |
| CN102585245A (en) * | 2012-01-13 | 2012-07-18 | 中科院广州化学有限公司 | High-dispersivity super-amphiphobic microsphere and self-cleaning epoxy resin paint prepared from same |
| CN103788264A (en) * | 2014-01-24 | 2014-05-14 | 中南大学 | Grafted acylamino hydroximic acid polymer and preparation method thereof |
-
2017
- 2017-07-20 CN CN201710596048.7A patent/CN107573451B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4209426A (en) * | 1979-05-29 | 1980-06-24 | American Home Products Corporation | Polypeptides related to somatostatin |
| CN102585245A (en) * | 2012-01-13 | 2012-07-18 | 中科院广州化学有限公司 | High-dispersivity super-amphiphobic microsphere and self-cleaning epoxy resin paint prepared from same |
| CN103788264A (en) * | 2014-01-24 | 2014-05-14 | 中南大学 | Grafted acylamino hydroximic acid polymer and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107573451A (en) | 2018-01-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10913987B2 (en) | Bacteria identification and antibiotic susceptibility profiling device | |
| Bhattacharjee et al. | Recent advances in sensor-based detection of toxic dyes for bioremediation application: a review | |
| Boedicker et al. | Detecting bacteria and determining their susceptibility to antibiotics by stochastic confinement in nanoliter droplets using plug-based microfluidics | |
| DE69630775T2 (en) | Electrochemiluminescence TEST | |
| Jorgensen et al. | Rapid penicillinase paper strip test for detection of beta-lactamase-producing Haemophilus influenzae and Neisseria gonorrhoeae | |
| CN104379760B (en) | Sample preparation for flow cytometry | |
| CN107209170A (en) | Electrochemical metabolic device for detecting activity | |
| Tomita et al. | Polymer-based chemical-nose systems for optical-pattern recognition of gut microbiota | |
| Buszewski et al. | Rapid microbiological diagnostics in medicine using electromigration techniques | |
| Needs et al. | Challenges in microfluidic and point-of-care phenotypic antimicrobial resistance tests | |
| CN107573451B (en) | A kind of beta-lactam antibiotic graft polymers and its application | |
| Thangadurai et al. | Separation and detection of certain β-lactam and fluoroquinolone antibiotic drugs by thin layer chromatography | |
| Thabit et al. | DETECTION OF EXTENDED-SPECTRUM-LACTAMASE ENZYMES (ESBLs) PRODUCED BY ESCHERICHIA COLI URINARY PATHOGENS AT ASSIUT UNIVERSITY HOSPITAL | |
| KR20210045879A (en) | Gram-negative bacterial detection composition comprising colistin conjugated with labeling material and method for detecting Gram-negative bacteria using thereof | |
| Riti et al. | Combining deep learning and droplet microfluidics for rapid and label-free antimicrobial susceptibility testing of colistin | |
| Butina et al. | Optotracing for selective fluorescence-based detection, visualization and quantification of live S. aureus in real-time | |
| Al-Mohammed et al. | Carbapenem resistance related with biofilm formation and pilin genes in clinical Pseudomonas aeruginosa isolates | |
| Mee'aad et al. | Detection of β-lactamases producing Methicillin-Resistant Staphylococcus aureus (MRSA) | |
| Otu et al. | Carriage of Plasmidmediated β-lactamase genes of Staphylococcus aureus isolates from Outpatients and Inpatients of UCTH, Calabar, Cross River State, Nigeria | |
| CN112945914A (en) | Fluorescence detection method for rapidly distinguishing gram types of bacteria | |
| Ugwu et al. | Antibiotic resistance patterns of Staphylococcus aureus isolated from nostrils of healthy human subjects in a southeastern Nigeria locality. | |
| Qureshi et al. | Status and distribution of MecA gene in hospitalized patients MRSA isolates | |
| AU597355B2 (en) | Method and apparatus for detection and quantitation of bacteria | |
| AZIZ et al. | Antimicrobial resistance;: comparison of Escherichia coli in different areas of lahore | |
| Jagad et al. | Comparison of three different methods to detect the production of β-lactamase enzyme by Sta phylococci |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |