CN102732474A - Compound for inhibiting mycobacterium tuberculosis, screening method and uses thereof - Google Patents
Compound for inhibiting mycobacterium tuberculosis, screening method and uses thereof Download PDFInfo
- Publication number
- CN102732474A CN102732474A CN2012101611450A CN201210161145A CN102732474A CN 102732474 A CN102732474 A CN 102732474A CN 2012101611450 A CN2012101611450 A CN 2012101611450A CN 201210161145 A CN201210161145 A CN 201210161145A CN 102732474 A CN102732474 A CN 102732474A
- Authority
- CN
- China
- Prior art keywords
- compound
- tyrosine phosphatase
- mycobacterium tuberculosis
- screening
- inhibition
- 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.)
- Pending
Links
- 150000001875 compounds Chemical class 0.000 title claims abstract description 81
- 238000012216 screening Methods 0.000 title claims abstract description 47
- 241000187479 Mycobacterium tuberculosis Species 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title claims abstract description 33
- 230000002401 inhibitory effect Effects 0.000 title abstract description 4
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims abstract description 64
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims abstract description 64
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 claims abstract description 58
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 claims abstract description 56
- 230000005764 inhibitory process Effects 0.000 claims abstract description 27
- 239000003814 drug Substances 0.000 claims abstract description 21
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 7
- 102000004190 Enzymes Human genes 0.000 claims abstract description 6
- 108090000790 Enzymes Proteins 0.000 claims abstract description 6
- 201000010099 disease Diseases 0.000 claims abstract description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 6
- 238000002360 preparation method Methods 0.000 claims abstract description 3
- ZQNOLGRKZRDRQO-XHSDSOJGSA-N bostrycin Chemical compound O[C@@H]1[C@@H](O)[C@@](C)(O)CC2=C1C(O)=C1C(=O)C=C(OC)C(=O)C1=C2O ZQNOLGRKZRDRQO-XHSDSOJGSA-N 0.000 claims description 30
- NCLWGURXHFTQGF-MGPLVRAMSA-N bostrycin Natural products COC1=CC(=O)c2c(O)c3C[C@H](O)[C@](C)(O)Cc3c(O)c2C1=O NCLWGURXHFTQGF-MGPLVRAMSA-N 0.000 claims description 30
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 8
- 230000009471 action Effects 0.000 claims description 8
- 241000894006 Bacteria Species 0.000 claims description 6
- 108090000623 proteins and genes Proteins 0.000 claims description 6
- 238000009585 enzyme analysis Methods 0.000 claims description 5
- 238000002189 fluorescence spectrum Methods 0.000 claims description 5
- 108010013043 Acetylesterase Proteins 0.000 claims description 4
- 241000588724 Escherichia coli Species 0.000 claims description 4
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 230000006798 recombination Effects 0.000 claims description 4
- 238000005215 recombination Methods 0.000 claims description 4
- 238000012795 verification Methods 0.000 claims description 4
- 238000001142 circular dichroism spectrum Methods 0.000 claims description 3
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 claims description 3
- 230000029087 digestion Effects 0.000 claims description 2
- 239000013604 expression vector Substances 0.000 claims description 2
- 239000012634 fragment Substances 0.000 claims description 2
- 230000000968 intestinal effect Effects 0.000 claims description 2
- 230000002503 metabolic effect Effects 0.000 claims description 2
- 244000005700 microbiome Species 0.000 claims description 2
- 230000009466 transformation Effects 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 17
- 229940079593 drug Drugs 0.000 abstract description 10
- 238000011282 treatment Methods 0.000 abstract description 4
- 201000008827 tuberculosis Diseases 0.000 abstract description 3
- 238000004458 analytical method Methods 0.000 abstract description 2
- 238000005516 engineering process Methods 0.000 abstract description 2
- 108700020796 Oncogene Proteins 0.000 abstract 1
- 102000043276 Oncogene Human genes 0.000 abstract 1
- 229940076850 Tyrosine phosphatase inhibitor Drugs 0.000 abstract 1
- 230000001355 anti-mycobacterial effect Effects 0.000 abstract 1
- 238000012258 culturing Methods 0.000 abstract 1
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 7
- 239000003112 inhibitor Substances 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 206010028980 Neoplasm Diseases 0.000 description 6
- 239000002512 suppressor factor Substances 0.000 description 6
- 239000000814 tuberculostatic agent Substances 0.000 description 6
- 238000013461 design Methods 0.000 description 5
- 201000011510 cancer Diseases 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 238000003149 assay kit Methods 0.000 description 3
- 239000003068 molecular probe Substances 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- IHIXIJGXTJIKRB-UHFFFAOYSA-N trisodium vanadate Chemical compound [Na+].[Na+].[Na+].[O-][V]([O-])([O-])=O IHIXIJGXTJIKRB-UHFFFAOYSA-N 0.000 description 3
- 102000004877 Insulin Human genes 0.000 description 2
- 108090001061 Insulin Proteins 0.000 description 2
- 241001049988 Mycobacterium tuberculosis H37Ra Species 0.000 description 2
- 208000008589 Obesity Diseases 0.000 description 2
- 108700019535 Phosphoprotein Phosphatases Proteins 0.000 description 2
- 102000045595 Phosphoprotein Phosphatases Human genes 0.000 description 2
- PLXBWHJQWKZRKG-UHFFFAOYSA-N Resazurin Chemical compound C1=CC(=O)C=C2OC3=CC(O)=CC=C3[N+]([O-])=C21 PLXBWHJQWKZRKG-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 210000001789 adipocyte Anatomy 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 210000002421 cell wall Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000013016 damping Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 230000005059 dormancy Effects 0.000 description 2
- 201000005577 familial hyperlipidemia Diseases 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 235000020824 obesity Nutrition 0.000 description 2
- 230000026731 phosphorylation Effects 0.000 description 2
- 238000006366 phosphorylation reaction Methods 0.000 description 2
- 239000002516 radical scavenger Substances 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 230000007923 virulence factor Effects 0.000 description 2
- 239000000304 virulence factor Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 229910000166 zirconium phosphate Inorganic materials 0.000 description 2
- WFJIVOKAWHGMBH-UHFFFAOYSA-N 4-hexylbenzene-1,3-diol Chemical compound CCCCCCC1=CC=C(O)C=C1O WFJIVOKAWHGMBH-UHFFFAOYSA-N 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 241001297689 Candidatus Glassbacteria Species 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 101000738772 Homo sapiens Receptor-type tyrosine-protein phosphatase beta Proteins 0.000 description 1
- 208000035150 Hypercholesterolemia Diseases 0.000 description 1
- 206010048723 Multiple-drug resistance Diseases 0.000 description 1
- 241000186359 Mycobacterium Species 0.000 description 1
- 241001646725 Mycobacterium tuberculosis H37Rv Species 0.000 description 1
- 108700035964 Mycobacterium tuberculosis HsaD Proteins 0.000 description 1
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 1
- 206010033645 Pancreatitis Diseases 0.000 description 1
- 108010009736 Protein Hydrolysates Proteins 0.000 description 1
- 102100037424 Receptor-type tyrosine-protein phosphatase beta Human genes 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 101710147108 Tyrosinase inhibitor Proteins 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 230000005784 autoimmunity Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000012292 cell migration Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000010307 cell transformation Effects 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 238000002983 circular dichroism Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 239000000287 crude extract Substances 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229940072185 drug for treatment of tuberculosis Drugs 0.000 description 1
- 239000003596 drug target Substances 0.000 description 1
- 238000003255 drug test Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 208000006575 hypertriglyceridemia Diseases 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- -1 isoxazole-salicylate compound Chemical class 0.000 description 1
- 210000001503 joint Anatomy 0.000 description 1
- 206010024627 liposarcoma Diseases 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000003032 molecular docking Methods 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 229910001453 nickel ion Inorganic materials 0.000 description 1
- 238000011275 oncology therapy Methods 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 238000002205 phenol-chloroform extraction Methods 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 230000004853 protein function Effects 0.000 description 1
- 239000003531 protein hydrolysate Substances 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000008521 reorganization Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 208000037803 restenosis Diseases 0.000 description 1
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 230000007727 signaling mechanism Effects 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention discloses a screening method of a compound for inhibiting mycobacterium tuberculosis. The method comprises the following steps: (1) establishing a screening model adopting tyrosine phosphatase as a target point: adopting genome DNA of tubercle bacillus H37 Rv as a template, adopting a PCR technology to clone tyrosine phosphatase gene, transforming host cells, culturing the transformant to obtain recombinant tyrosine phosphatase from the culture, carrying out enzyme activity analysis, and establishing the screening model adopting tyrosine phosphatase as the target point; and (2) adopting the screening model established in the step (1) to screen a compound providing inhibition activity for the tyrosine phosphatase. With the method of the present invention, the tyrosine phosphatase inhibitor with anti-mycobacterium tuberculosis effect can be rapidly and efficiently screened. In addition, the present invention further discloses a compound screened by using the screening method, and uses of the compound in preparations of drugs for treatments of diseases, wherein the inducement or one of the inducements of the diseases is the tyrosine phosphatase.
Description
Technical field
The present invention relates to a kind of screening method of mycobacterium tuberculosis compound, the compound that adopts said screening method screening gained inhibition mycobacterium tuberculosis and purposes of this compound of suppressing.
Background technology
White plaque is the communicable disease that is caused by mycobacterium tuberculosis (Mycobacterium tuberculosis).According to World Health Organization's statistics, the whole world has 1/3 population to infect tubercule bacillus approximately, and 8,800,000 new cases are arranged every year approximately, and 2,000,000 people are dead because of white plaque.At present the tuberculotherapy medicine needs life-time service, and is very big to the injury of human body, and infectious bacteria can hide and become the dormancy bacterium in the focus midium or long term of host's caseate, in case the defence immunity function lowly get final product " reviving " and falls ill again.Appearance of multiple-drug resistance tuberculosis mycobacterium and tubercule bacillus have become the significant problem in tuberculosis prophylaxis and the therapeutic process at patient's intravital " hold and stay " state at present.Therefore, seek new drug target, set up new medicaments sifting model, it is very crucial to develop the powerful low toxicity antitubercular agent that can control the multi-drug resistant tubercule bacillus and eradicate the dormancy bacterium.
The method of seeking novel antitubercular agent at present is limited to drug sensitive test mostly, but mycobacterium tuberculosis is slow owing to the speed of growth, and a drug sensitive test cycle reaches 3-4 week, is inappropriate for extensive screening, the also difficult newtype drug of finding.
The tradition antitubercular agent synthesizes through the inhibition cell walls or bacterial growth is killed MTB, but can produce the MTB selection pressure simultaneously.The medicine of target secretor type virulence factor efficiently solves this problem, produces the perviousness obstacle when also having avoided medicine through cell walls.Patient (like HIV the infected) who suffers damage for immunity system and excessive risk infect the medical personnel of TB, and these medicines also can reduce the infection probability.Infection can also be effectively removed in this type of suppressor factor and traditional microbiotic coupling, reduces the MTB resistance and produces.MPTPA/mPTPB is the secretor type virulence factor of MTB, is the suppressor factor that drug targets filters out much has application prospect with mPTPA/mPTPB, is indicating the alternative of following TB treatment.The potentiality that the suppressor factor of at present a lot of mPTPA/mPTPB makes further progress.Newfound have dibit to put active isoxazole-salicylate compound to have cytoactive, in clear its treatment effect of the scavenger cell internal evidence of infecting MTB.Compare with contrast, subdued the growth of MTB90% in the scavenger cell.It is that its greater activity reaches optionally architecture basics that its unique dibit point combines to suppress model, and this structure also provides thinking for later design novel inhibitors.But more suppressor factor has hindered their further optimization owing to lack crystalline structure information.As the standard of judging medicine, combining mixture to carry out crystalline structure research and zootype experiment with target to suppressor factor is present urgent need.
Tyrosine phosphatase is the tyrosine phosphatase ester bond on the protein hydrolysate substrate specifically, dephosphorylate, thus regulate the enzyme of this protein function.The phosphorylation of protein-tyrosine residue plays an important role for regulating the physiologic response process, for example cell proliferation and differentiation, cell migration, activated immune cell and apoptosis etc.Therefore; If the tyrosine phosphorylation effect imbalance in the cell can cause serious consequence, comprise to cause that autoimmunity, improper metabolism reply and cell transformation etc. that the consequent is the generation of numerous disease; Comprise: cancer, mellitus and autoimmunization defective.There is report PTPB specific inhibitor to be expected to improve the susceptibility of Regular Insulin, treats diabetes B, insulin resistant and obesity effectively.This just points out us; Tyrosinase inhibitor possibly be unusual effect also to be arranged aspect the disease that causes of mPTPB in the treatment inducement; Comprise: mellitus, obesity, hyperlipemia, hypertriglyceridaemia, hypercholesteremia, low HDL levels, atherosclerosis, vascular restenosis, inflammatory bowel, pancreatitis, adipocyte tumour, adipocyte cancer, liposarcoma, hyperlipemia, tumour, cancer, autoimmune disease etc.
White plaque is as global public health problem, and through using for reference cancer therapy drug achievement in research and research thinking, the method in conjunction with the signaling mechanism and the inhibitor screening of phosphoprotein phosphatase in the cancer cells helps to quicken new antitubercular agent and appears on the market.
Summary of the invention
The objective of the invention is to overcome the weak point of prior art and a kind of screening method that suppresses the mycobacterium tuberculosis compound is provided; The present invention simultaneously also provides the compound that adopts said screening method to screen the inhibition mycobacterium tuberculosis that obtains, and the purposes of said compound.
For realizing above-mentioned purpose, the technical scheme that the present invention takes is: a kind of screening method that suppresses the mycobacterium tuberculosis compound may further comprise the steps:
(1) setting up with the tyrosine phosphatase is the screening model of target spot: DNA is a template with tubercule bacillus H37Rv pnca gene group; PCR clone tyrosine phosphatase gene; Transform host cell, the culture transformation body obtains the recombination tyrosine Phosphoric acid esterase from culture; Carry out enzyme analysis alive, setting up with the tyrosine phosphatase is the screening model of target spot;
(2) the screening model screening that utilizes step (1) to be set up has the active compound of inhibition to tyrosine phosphatase.
Preferred implementation as the screening method of inhibition mycobacterium tuberculosis compound according to the invention; The host cell that adopts in the said step (1) is intestinal bacteria,, is connected with pET-28a (+) carrier through double digestion with the purpose fragment of expression vector; Be converted into e. coli bl21; Induce through IPTG, obtain highly-soluble and express, obtain the recombination tyrosine Phosphoric acid esterase.
As the preferred implementation of the screening method of inhibition mycobacterium tuberculosis compound according to the invention, described IPTG final concentration is 0.1～0.2mM, and inducing temperature is 18～37 ℃.
Preferred implementation as the screening method of inhibition mycobacterium tuberculosis compound according to the invention; Adopt tyrosine phosphatase enzyme reagent kit (RediPlate.96
TyrosinePhosphatase Assay Kit in the said step (1); Molecular Probes Inc) carries out enzyme analysis alive to the reorganization tyrosine phosphatase.
Simultaneously, the present invention also provides a kind of compound that suppresses mycobacterium tuberculosis, and said compound adopts above-mentioned screening method screening to obtain.The action target spot of said compound is a tyrosine phosphatase.The compound that obtains that screens has against mycobacterium tuberculosis effect, can effectively suppress the tyrosine phosphatase enzymic activity.Said compound called after Bostrycin.
As the preferred implementation of the compound of inhibition mycobacterium tuberculosis according to the invention, said compound called after Bostrycin, the molecular structural formula of said compound is:
As the preferred implementation of the compound of inhibition mycobacterium tuberculosis according to the invention, said compound is the natural metabolic activity product of marine microorganism.
As the preferred implementation of the compound of inhibition mycobacterium tuberculosis according to the invention, the minimum concentration that said compound suppresses mycobacterium tuberculosis is 7.5 μ g/mL, and this result adopts micro-resazurin method to measure gained.
As the preferred implementation of the compound of inhibition mycobacterium tuberculosis according to the invention, the action target spot of said compound is a tyrosine phosphatase.The verification method of the action target spot of said compound may further comprise the steps:
(1) the modelling verification compound that utilizes step in the claim 1 (1) to be set up is lived to the enzyme of tyrosine phosphatase and is suppressed active;
(2) utilize Docking software to carry out information biology prediction, compound and tyrosine phosphatase enzyme interacting;
(3) utilize fluorescence spectrum checking, compound and tyrosine phosphatase enzyme interacting;
(4) utilize circular dichroism spectrum checking, compound and tyrosine phosphatase enzyme interacting.
The present invention also provides the compound that suppresses mycobacterium tuberculosis as stated to be used for treating the purposes of medicine that one of inducement or inducement are the diseases of tyrosine phosphatase in preparation.
The screening method of inhibition mycobacterium tuberculosis compound according to the invention; At first set up with the tyrosine phosphatase is the screening model of target spot; The screening model of setting up can fast, efficiently screen the tyrosine phosphatase inhibitors with Killing Mycobacterium Tuberculosis effect; Reduce workload, accelerate working efficiency, improve the probability that obtains positive experimental result.The compound that adopts screening method screening according to the invention to obtain has against mycobacterium tuberculosis effect, is action target spot with the tyrosine phosphatase, can effectively suppress the tyrosine phosphatase enzymic activity, has bigger potential applicability in clinical practice and potentiality.
Description of drawings
Fig. 1 suppresses the molecular structural formula of the compound of mycobacterium tuberculosis for the present invention.
Fig. 2 is that tyrosine phosphatase is expressed figure in the mycobacterium tuberculosis H37Rv genome in E.coli BL21 (PlysS);
Among Fig. 2, M is standard protein molecular weight maker, and 1 is the recombinant protein crude extract, and 2 is purified recombinant albumen.
Fig. 3 is different concns tyrosine phosphatase enzyme figure alive;
Among Fig. 3, from a ~ e, the tyrosine phosphatase enzyme concn is 10 times of increases.
Fig. 4 is based on the active inhibitor screening figure of tyrosine phosphatase;
Among Fig. 4, No. 1 is blank, be for No. 2 former sodium vanadate over against photograph, No. 3 negative contrasts, 4 ~ No. 29 is the compound that is screened.
Fig. 5 is based on the active inhibitor screening figure of tyrosine phosphatase;
Among Fig. 5, No. 30 is blank, be for No. 31 former sodium vanadate over against photograph, No. 32 negative contrasts, 33 ~ No. 59 is the compound that is screened.
Fig. 6 suppresses the design sketch of mycobacterium tuberculosis H37Ra for part of compounds;
Among Fig. 6,37 representation compound Bostrycin, 38 ~ 41 represent the verivate of Bostrycin.
Fig. 7 suppresses the design sketch of mycobacterium tuberculosis H37Ra for part of compounds;
Among Fig. 7,42 ~ 44 represent the verivate of Bostrycin, and 45 represent medicament benemicin, and 46 represent the solvent DMSO of dissolved compound.
Fig. 8 is Bostrycin and mPTPB interaction synoptic diagram
The interactional fluorescence emission spectrogram of the compound Bostrycin of different concns ratio and tyrosine phosphatase mPTPB when Fig. 9 is 25 ℃;
Among Fig. 9, a:0; B:0.25; C:0.5; D:0.75; E:1; F:1.25; G:1.5.
The DMSO of different concns ratio and the interactional fluorescence emission spectrum of tyrosine phosphatase mPTPB when Figure 10 is 25 ℃;
Among Figure 10, a:0; B:0.25; C:0.5; D:0.75; E:1; F:1.25; G:1.5.
Figure 11 is the compound Bostrycin and the interactional CD spectrogram of tyrosine phosphatase mPTPB of different concns ratio;
Among Figure 11, a:0; B:0.5; C:1; D:1.5.
Figure 12 is the methyl alcohol and the interactional CD spectrogram of tyrosine phosphatase mPTPB of different concns ratio;
Among Figure 12, a:0; B:0.5; C:1; D:1.5.
Embodiment
The object of the invention, technical scheme and advantage will combine accompanying drawing and specific embodiment that the present invention is described further below for better explaining.
The foundation of screening model in the screening method of embodiment 1 inhibition mycobacterium tuberculosis compound
Present embodiment has extracted tubercule bacillus H37Rv genomic dna with the phenol chloroform extraction method; And be template; PCR clone mptpb gene; It is efficiently expressed in E.coli BL21 (PlysS), see shown in the accompanying drawing 2, utilize the nickel ion affinity chromatograph column purification to obtain mPTPB albumen; Utilize tyrosine phosphatase enzyme reagent kit (RediPlate.96
Tyrosine Phosphatase Assay Kit; Molecular Probes Inc) carries out enzyme analysis alive, and having set up with the tyrosine phosphatase is the screening model of target spot.The tyrosine phosphatase enzyme work of different concns is shown in accompanying drawing 3.
The application of screening model in the screening method of embodiment 2 inhibition mycobacterium tuberculosis compounds
1, inhibitor screening
What utilize embodiment 1 foundation is the screening model of target spot with the tyrosine phosphatase; Compound is suppressed tyrosine phosphatase mPTPB enzymic activity to be analyzed; Adopt tyrosine phosphatase enzyme reagent kit (RediPlate.96
Tyrosine Phosphatase Assay Kit; Molecular Probes; Inc); Compound and the verivate thereof of 84 kinds of separation from thalassiomycetes screened, and wherein 42 kinds of compounds have the inhibition activity of enzyme, and drug sensitive test and MIC test that H37Ra carries out are found have 39 kinds to have inhibition zone.Wherein, compound Bostrycin (molecular structural formula is seen shown in the accompanying drawing 1) and verivate thereof, inhibitory enzyme activity and inhibition zone have a stronger dependency, the action target spot that this medicine is described is mPTPB.
The used inhibition of present embodiment mPTPB enzymic activity over against according to being Trisodium vanadate (Na
3VO
4), test design sees table 1 for details.
The design of table 1 screening novel anti tuberculosis drug test
The selection result figure based on the active suppressor factor of tyrosine phosphatase sees to attach shown in the Figure 4 and 5.
2, the drug sensitive test paper method is measured the compound inhibition zone
Every substratum pastes 5 drug sensitive test papers with even interval after evenly being coated with mycobacterium tuberculosis, add the 5ug compound respectively, places 37 ℃ and cultivates for 3～4 weeks, afterwards observed and recorded inhibition zone size.The gained result sees shown in accompanying drawing 6 and 7.
3, micro-resazurin method is measured the minimum inhibitory concentration MIC value that compound suppresses mycobacterium tuberculosis
(1) every hole adds 7H9 substratum 100 μ L in aseptic 96 orifice plates (1～10 hole), and the 1st hole adds the antitubercular agent stoste 100 μ L of suitable dilution, double serial dilution to the 8 holes;
(2) the fresh culture bacterium of 2 10%Tween-80 saline water of adding and 2～3 all cell ages bottom the glass bacteria grinder; Be ground to and be cheese appearance; Be diluted to the turbidity (1mg/mL) of No. 1 Maxwell opacity tube with saline water; Inoculate 100 μ L with every hole, 1～9 hole after 1: 20 times of dilution of 7H9 substratum, the 10th hole is the substratum contrast;
(3) with no mycoderm shrouding, put into wet box, cultivate 5d for 37 ℃; Add filtration sterilization 0.1g/L resazurin colour developing liquid 30 μ L to the 9 holes on the 6th day, continue incubation 24h, red like the 9th hole (not adding medicine) pulverize; Then add with the amount resazurin and develop the color liquid, write down colour-change behind the 24h to other each holes; Like the 9th hole still is blue, observes at the 9th, 11 day respectively, and color becomes pink colour from blueness and promptly indicates bacterial growth, to keep blue minimum medicine.
The action target spot of embodiment 3 checking compound Bostrycins is a tyrosine phosphatase
1, Docking analyzes
Utilize the Sybyl-x1.1 computed in software to go out RMSD=0.5335, be worth lessly, possibly compare near actual butt joint situation, explain when Bostrycin combines with mPTPB, its position concerns that the position when mPTPB combines with part in the 20Z5 crystal concerns suitable.With original part in the 20Z5 protein structures and average mark after 20Z5 docks: 12.0515.Best result is 4.41 to use Bostrycin to dock afterwards with 20Z5, and Bostrycin and marking after 20Z5 docks are all less than former part and average mark after 20Z5 docks.The binding ability that Bostrycin and acceptor 20Z5 are described possibly be weaker than former part.But its gap has only about 3 times.Reason possibly be that former part has bigger space structure, and has flexible preferably.And the space structure of Bostrycin is less; And have stronger rigidity, structure is not easy to be distorted to cater to the needs of acceptor pocket.
Can be found out that by Fig. 8 bostrycin can interact with the mPTPB crystal, its main binding site is Ser57, and Asp165 and Arg166 compare with the site plan of mPTPB self part, and its binding site is all more similar with bond angle.Report; 160 ~ 166 amino acids can form a P-Loop, and this structure is extremely important for the formation of active pocket, and bostrycin combines with Asp165 and Arg166; Exactly broken this structure, this possibly also be the major reason that bostrycin can suppress the mPTPB enzymic activity.
2, fluorescent spectroscopy
The degree of fluorescent quenching is relevant with the binding ability that adds Bostrycin and mPTPB, and bonding strength is big more, and the amplitude that fluorescence reduces is big more.Take 1cm trace quartz colorimetric utensil earlier, add the mPTPB solution that does not contain Bostrycin, on XRF, excite with 230nm; 250-800nm scanning detects this fluorescence intensity of solution, keeps the interior mPTPB protein concentration (0.02 μ g/ μ L) of cuvette constant subsequently; Add Bostrycin gradually, making its concentration is 0.005 μ g/ μ L, 0.01 μ g/ μ L; 0.015 μ g/ μ L, 0.02 μ g/ μ L, 0.025 μ g/ μ L; 0.03 μ g/ μ L continues to detect the fluorescence curve that mPTPB produces with the Bostrycin concentration change.The Bostrycin of different concns ratio sees respectively shown in accompanying drawing 9 and 10 with the DMSO and the interactional fluorescence emission spectrum of mPTPB of interactional fluorescence emission spectrum of mPTPB and different concns ratio in the time of 25 ℃.
3, circular dichroism spectrum analysis
The CD figure that on JASCO J-810 circular dichroism spectrometer, measures damping fluid PBS measures blank and deducts as baseline as blank.Be mixed with the mPTPB solution that concentration is 0.2ug/uL with measuring damping fluid (PBS), the CD spectrogram in the 200-800nm scope behind the Bostrycin of measurement mPTPB and adding different concns thereof.What the present invention taked is the method for Bostrycin titration mPTPB, and promptly mPTPB concentration is constant, according to certain mass than the concentration that increases Bostrycin till considerable change does not take place in the CD of mixture collection of illustrative plates.Wherein Bostrycin adopts methyl alcohol as solvent; Be mixed with concentration and be about the suspension-s of 100mg/mL; And ultrasonic hydrotropy, adding 0.5uLBostrycin at every turn, the concentration ratio that makes Bostrycin and mPTPB is 0.5,1,1.5; After Bostrycin joins mPTPB solution, directly sweep spectrum and do not pass through other incubation time.The cuvette model is 1cm, and accumulation scanning is 3-4 time under the room temperature, and speed is 500nm/min.The compound Bostrycin of different concns ratio is seen respectively shown in accompanying drawing 11 and 12 with the methyl alcohol and the interactional CD spectrogram of tyrosine phosphatase mPTPB of interactional CD spectrogram of tyrosine phosphatase mPTPB and different concns ratio.
Last institute should be noted that; Above embodiment is only in order to technical scheme of the present invention to be described but not to the restriction of protection domain of the present invention; Although the present invention has been done detailed description with reference to preferred embodiment; Those of ordinary skill in the art should be appreciated that and can make amendment or be equal to replacement technical scheme of the present invention, and do not break away from the essence and the scope of technical scheme of the present invention.
Claims (10)
1. a screening method that suppresses the mycobacterium tuberculosis compound is characterized in that, may further comprise the steps:
(1) setting up with the tyrosine phosphatase is the screening model of target spot: DNA is a template with tubercule bacillus H37Rv pnca gene group; PCR clone tyrosine phosphatase gene; Transform host cell, the culture transformation body obtains the recombination tyrosine Phosphoric acid esterase from culture; Carry out enzyme analysis alive, setting up with the tyrosine phosphatase is the screening model of target spot;
(2) the screening model screening that utilizes step (1) to be set up has the active compound of inhibition to tyrosine phosphatase.
2. the screening method of inhibition mycobacterium tuberculosis compound as claimed in claim 1 is characterized in that, the host cell that adopts in the said step (1) is intestinal bacteria; With the purpose fragment of expression vector through double digestion; Be connected with pET-28a (+) carrier, be converted into e. coli bl21, induce through IPTG; Obtain highly-soluble and express, obtain the recombination tyrosine Phosphoric acid esterase.
3. the screening method of inhibition mycobacterium tuberculosis compound as claimed in claim 1 is characterized in that, adopts tyrosine phosphatase enzyme reagent kit counterweight group tyrosine phosphatase to carry out enzyme analysis alive in the said step (1).
4. a compound that suppresses mycobacterium tuberculosis is characterized in that, said compound adopts the method screening in the claim 1 to obtain.
5. the compound of inhibition mycobacterium tuberculosis as claimed in claim 3 is characterized in that, said compound called after Bostrycin, and the molecular structural formula of said compound is:
6. like the compound of claim 4 or 5 described inhibition mycobacterium tuberculosis, it is characterized in that said compound is the natural metabolic activity product of marine microorganism.
7. like the compound of claim 4 or 5 described inhibition mycobacterium tuberculosis, it is characterized in that the minimum concentration that said compound suppresses mycobacterium tuberculosis is 7.5 μ g/mL.
8. one kind like claim 4 or 5 described compounds, it is characterized in that the action target spot of said compound is a tyrosine phosphatase.
9. a compound as claimed in claim 8 is characterized in that, the verification method of the action target spot of said compound may further comprise the steps:
(1) the modelling verification compound that utilizes step in the claim 1 (1) to be set up is lived to the enzyme of tyrosine phosphatase and is suppressed active;
(2) utilize Docking software to carry out information biology prediction, compound and tyrosine phosphatase enzyme interacting;
(3) utilize fluorescence spectrum checking, compound and tyrosine phosphatase enzyme interacting;
(4) utilize circular dichroism spectrum checking, compound and tyrosine phosphatase enzyme interacting.
10. one kind is used for treating the purposes of medicine that one of inducement or inducement are the diseases of tyrosine phosphatase like claim 4 or 5 said compounds in preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012101611450A CN102732474A (en) | 2012-05-21 | 2012-05-21 | Compound for inhibiting mycobacterium tuberculosis, screening method and uses thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012101611450A CN102732474A (en) | 2012-05-21 | 2012-05-21 | Compound for inhibiting mycobacterium tuberculosis, screening method and uses thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102732474A true CN102732474A (en) | 2012-10-17 |
Family
ID=46988777
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012101611450A Pending CN102732474A (en) | 2012-05-21 | 2012-05-21 | Compound for inhibiting mycobacterium tuberculosis, screening method and uses thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102732474A (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103446079A (en) * | 2013-06-26 | 2013-12-18 | 中山大学 | Application of Bostrycin to preparation of protein tyrodine phosphatase (PTP) bonding agents or inhibitors |
| CN103694209A (en) * | 2013-06-07 | 2014-04-02 | 中山大学 | Marine fungus-derived acetophenone compounds and preparation method and application thereof |
| CN106620831A (en) * | 2016-12-22 | 2017-05-10 | 石佳明 | Dressing for caring skin ulcer |
| CN108409816A (en) * | 2018-04-13 | 2018-08-17 | 扬州工业职业技术学院 | A method of the separating flavone class compound from sweet potato leaves |
| CN108498560A (en) * | 2018-04-13 | 2018-09-07 | 扬州工业职业技术学院 | A kind of sweet potato leaf flavone extract and the preparation method and application thereof |
| CN108586553A (en) * | 2018-04-13 | 2018-09-28 | 扬州工业职业技术学院 | A kind of novel flavone sugar glycosides compound and its application as MptpB inhibitor |
| CN109836396A (en) * | 2017-11-29 | 2019-06-04 | 扬州蓝色生物医药科技有限公司 | A kind of novel thiazole sulfamide compound and its application as antituberculotic |
| CN109836395A (en) * | 2017-11-29 | 2019-06-04 | 扬州蓝色生物医药科技有限公司 | A kind of o-trifluoromethyl phenoxy acetamide base thiazole sulfonamide MptpB inhibitor |
| CN111658629A (en) * | 2020-07-07 | 2020-09-15 | 中山大学 | Application of Fusariulin M and derivatives and pharmaceutically acceptable salts thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007093557A1 (en) * | 2006-02-13 | 2007-08-23 | Laboratoires Serono S.A. | Sulfonamide derivatives for the treatment of bacterial infections |
-
2012
- 2012-05-21 CN CN2012101611450A patent/CN102732474A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007093557A1 (en) * | 2006-02-13 | 2007-08-23 | Laboratoires Serono S.A. | Sulfonamide derivatives for the treatment of bacterial infections |
Non-Patent Citations (3)
| Title |
|---|
| BRIAN BEAGLEY ET AL.: "Revision of the Absolute Configuration of (-)-Bostrycin", 《J. CHEM. SOC.》 * |
| LIHUA DONG ET AL.: "Theoretical studies on the interaction of biphenyl inhibitors with Mycobacterium tuberculosis protein tyrosine phosphatase MptpB", 《J MOL MODEL》 * |
| 黄宇虹等: "bostrycin 的抗结核菌活性及对细菌基因表达的影响", 《中国新药杂志》 * |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103694209A (en) * | 2013-06-07 | 2014-04-02 | 中山大学 | Marine fungus-derived acetophenone compounds and preparation method and application thereof |
| CN103694209B (en) * | 2013-06-07 | 2015-10-28 | 中山大学 | The acetophenone compounds in one class thalassiomycetes source and preparation method thereof and application |
| CN103446079A (en) * | 2013-06-26 | 2013-12-18 | 中山大学 | Application of Bostrycin to preparation of protein tyrodine phosphatase (PTP) bonding agents or inhibitors |
| CN106620831A (en) * | 2016-12-22 | 2017-05-10 | 石佳明 | Dressing for caring skin ulcer |
| CN109836395A (en) * | 2017-11-29 | 2019-06-04 | 扬州蓝色生物医药科技有限公司 | A kind of o-trifluoromethyl phenoxy acetamide base thiazole sulfonamide MptpB inhibitor |
| CN109836395B (en) * | 2017-11-29 | 2020-08-04 | 扬州蓝色生物医药科技有限公司 | O-trifluoromethyl phenoxyacetamido thiazole sulfonamide MptpB inhibitor |
| CN109836396B (en) * | 2017-11-29 | 2020-08-04 | 扬州蓝色生物医药科技有限公司 | Novel thiazole sulfonamide compound and application thereof as antituberculosis drug |
| CN109836396A (en) * | 2017-11-29 | 2019-06-04 | 扬州蓝色生物医药科技有限公司 | A kind of novel thiazole sulfamide compound and its application as antituberculotic |
| CN108586553A (en) * | 2018-04-13 | 2018-09-28 | 扬州工业职业技术学院 | A kind of novel flavone sugar glycosides compound and its application as MptpB inhibitor |
| CN108409816B (en) * | 2018-04-13 | 2019-12-10 | 扬州工业职业技术学院 | Method for separating flavonoid compounds from sweet potato leaves |
| CN108586553B (en) * | 2018-04-13 | 2019-12-13 | 扬州工业职业技术学院 | Novel flavone glycoside compound and application thereof as MptpB inhibitor |
| CN108498560A (en) * | 2018-04-13 | 2018-09-07 | 扬州工业职业技术学院 | A kind of sweet potato leaf flavone extract and the preparation method and application thereof |
| CN108409816A (en) * | 2018-04-13 | 2018-08-17 | 扬州工业职业技术学院 | A method of the separating flavone class compound from sweet potato leaves |
| CN111658629A (en) * | 2020-07-07 | 2020-09-15 | 中山大学 | Application of Fusariulin M and derivatives and pharmaceutically acceptable salts thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102732474A (en) | Compound for inhibiting mycobacterium tuberculosis, screening method and uses thereof | |
| Huentelman et al. | Structure-based discovery of a novel angiotensin-converting enzyme 2 inhibitor | |
| CN107190006A (en) | A kind of sgRNA of targeting IGF IR genes and its application | |
| León et al. | Abyssomicin 2 reactivates latent HIV-1 by a PKC-and HDAC-independent mechanism | |
| CN107075473A (en) | The wherein cell mass of the removed induction of neoblast, its purposes and preparation method thereof | |
| CN102427814A (en) | Kinase protein binding inhibitors | |
| Fung et al. | Unbiased screening of marine sponge extracts for anti-inflammatory agents combined with chemical genomics identifies girolline as an inhibitor of protein synthesis | |
| CN102911040A (en) | Sesquiterpenoids from marine fungi source as well as preparation method and application thereof | |
| CN104120164A (en) | Specific fluorescence probe substrates of human carboxylesterase 2 and application thereof | |
| CN105879030A (en) | Synergistic drug compound for treating tumor and preparation method thereof | |
| CN104101714B (en) | To suppress the screening anti-tumor medicine method that Smurf1 ubiquitination degraded RhoB is target spot | |
| Yang et al. | TGF‑β/Smad3 pathway enhances the cardio‑protection of S1R/SIPR1 in in vitro ischemia‑reperfusion myocardial cell model | |
| CN105617381B (en) | The new application of histon deacetylase (HDAC) inhibitor treatment β subfamily herpesviral | |
| Li et al. | ACE2 suppresses the inflammatory response in LPS-induced porcine intestinal epithelial cells via regulating the NF-κB and MAPK pathways | |
| CN107281172A (en) | Application of the melbine in the medicine for preparing cervical carcinoma | |
| CN104193686B (en) | N-Galla Turcica (Galla Helepensis) amide-pyrimidine radicals benzsulfamide analog derivative and its preparation method and application | |
| KR20230152715A (en) | Composition for treatment of EBV-related disease or condition | |
| CN100346781C (en) | Use of okadaic acid in preparation of medicine for resisting glaucoma operation scar | |
| CN107058476A (en) | Applications of the EXT1 in diagnosing cancer of liver treatment | |
| RU2190220C1 (en) | Method for finding tuberculosis micobacteria | |
| CN110772503A (en) | Application of 2-methoxy-6-acetyl-7-methyl walnut ketone | |
| CN116829189A (en) | SARMS compound containing ester aromatic propionamide and application of metabolite thereof in preparation of anti-novel coronavirus drugs | |
| Meyers et al. | Crosstalk between epidermal growth factor receptor and P-glycoprotein in actinomycin D-resistant Chinese hamster lung cells | |
| TWI453028B (en) | Radiosensitizer compositions comprising schisandra chinensis (turcz.) baill and methods for use | |
| CN104689341A (en) | Application of PP2A antagonist in preparation of medicines for treating acute liver injury |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20121017 |