CN109771415A - A kind of micromolecular inhibitor containing benzodioxole and the application in inhibition ornithine decarboxylase (ODC) - Google Patents
A kind of micromolecular inhibitor containing benzodioxole and the application in inhibition ornithine decarboxylase (ODC) Download PDFInfo
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- CN109771415A CN109771415A CN201910185281.5A CN201910185281A CN109771415A CN 109771415 A CN109771415 A CN 109771415A CN 201910185281 A CN201910185281 A CN 201910185281A CN 109771415 A CN109771415 A CN 109771415A
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- odc
- micromolecular inhibitor
- benzodioxole
- ornithine decarboxylase
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- 102000052812 Ornithine decarboxylases Human genes 0.000 title claims abstract description 78
- 108700005126 Ornithine decarboxylases Proteins 0.000 title claims abstract description 78
- 239000003112 inhibitor Substances 0.000 title claims abstract description 48
- FTNJQNQLEGKTGD-UHFFFAOYSA-N 1,3-benzodioxole Chemical compound C1=CC=C2OCOC2=C1 FTNJQNQLEGKTGD-UHFFFAOYSA-N 0.000 title claims abstract description 12
- 230000005764 inhibitory process Effects 0.000 title description 9
- 239000003814 drug Substances 0.000 claims abstract description 11
- 229940079593 drug Drugs 0.000 claims abstract description 10
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- 208000015181 infectious disease Diseases 0.000 claims abstract description 5
- 238000002360 preparation method Methods 0.000 claims abstract description 5
- 230000002401 inhibitory effect Effects 0.000 claims description 12
- 238000011282 treatment Methods 0.000 abstract description 7
- FAWGDZUNQZJISX-UHFFFAOYSA-N heptanehydrazide Chemical compound CCCCCCC(=O)NN FAWGDZUNQZJISX-UHFFFAOYSA-N 0.000 abstract description 2
- 230000000694 effects Effects 0.000 description 17
- 238000000034 method Methods 0.000 description 16
- 108090000623 proteins and genes Proteins 0.000 description 14
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- 238000001514 detection method Methods 0.000 description 8
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- 239000000758 substrate Substances 0.000 description 7
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- AHLPHDHHMVZTML-UHFFFAOYSA-N Orn-delta-NH2 Natural products NCCCC(N)C(O)=O AHLPHDHHMVZTML-UHFFFAOYSA-N 0.000 description 6
- UTJLXEIPEHZYQJ-UHFFFAOYSA-N Ornithine Natural products OC(=O)C(C)CCCN UTJLXEIPEHZYQJ-UHFFFAOYSA-N 0.000 description 6
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- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 4
- 150000003384 small molecules Chemical class 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- ATHGHQPFGPMSJY-UHFFFAOYSA-N spermidine Chemical compound NCCCCNCCCN ATHGHQPFGPMSJY-UHFFFAOYSA-N 0.000 description 4
- PFNFFQXMRSDOHW-UHFFFAOYSA-N spermine Chemical compound NCCCNCCCCNCCCN PFNFFQXMRSDOHW-UHFFFAOYSA-N 0.000 description 4
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- VLCYCQAOQCDTCN-UHFFFAOYSA-N eflornithine Chemical compound NCCCC(N)(C(F)F)C(O)=O VLCYCQAOQCDTCN-UHFFFAOYSA-N 0.000 description 3
- 239000001963 growth medium Substances 0.000 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 description 3
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- UQGFMSUEHSUPRD-UHFFFAOYSA-N disodium;3,7-dioxido-2,4,6,8,9-pentaoxa-1,3,5,7-tetraborabicyclo[3.3.1]nonane Chemical compound [Na+].[Na+].O1B([O-])OB2OB([O-])OB1O2 UQGFMSUEHSUPRD-UHFFFAOYSA-N 0.000 description 2
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Landscapes
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The present invention provides a kind of micromolecular inhibitor containing benzodioxole, the micromolecular inhibitor is N'- ({ 6- nitro -1,3- benzodioxole -5- base } methylene) spiral shell [2.3] hexane -1- carbohydrazide, structural formula are as follows:The micromolecular inhibitor containing benzodioxole is being inhibited the application on ornithine decarboxylase (ODC) by the present invention, and the application in preparation tumor, and the application in preparation treatment cause pathogeny imcrobe infection drug, and obtain remarkable result.
Description
Technical field
The present invention relates to ornithine decarboxylase (Ornithine decarboxylase；ODC micromolecular inhibitor) and its
Using, and in particular to the micromolecular inhibitor of source of people ornithine decarboxylase and its application in inhibition and killing tumor cell.
Background technique
Protein is one of main constituents of organism, is the main matter for completing various vital movements.Various
In protein, protease is most important to vital movement, the intracorporal biochemical reaction process of almost all creatures all by protease into
Row catalysis.There is the activity of various protease stringent regulatory mechanism to cause once its regulatory mechanism goes wrong in organism
The hyperactivity of protease, too low or complete deactivation, can all cause corresponding various diseases.Therefore, it is adjusted by drug
The activity for controlling protease makes it restore and be maintained at normal level, has very important theory significance and realistic meaning.With knot
Drug design based on structure, is the very important means designed using protein as the drug of target.
Polyamines (polyamines) is a kind of positively charged cation micro molecule generated from amino acid metabolism, in all lifes
All exist in object, it is all indispensable to cell growth, differentiation, survival and natural biological function etc..The more positive charges of polyamines band
Characteristic, enable them to make by forming electrostatic with negatively charged large biological molecule (DNA, RNA, protein, cell membrane etc.)
With to regulate and control very extensive biological process, including chromosome knob is configured to, DNA is synthesized and stabilization, DNA replication dna, transcription
It is generated with translation, protein phosphorylation, ribosomes, ion channel and regulation, the radicals scavenging of film surface receptor etc..Natural
There are many kinds of polyamines.In mammals, there are three types of naturally occurring, i.e. putrescine (putrescine), spermidine
(spermidine), spermine (spermine), they are essential to mammal normal growth and development.Since polyamines has
Important biological function, Intracellular levels are by stringent regulation.It is more such as tumour cell in the cell quickly bred
Amine level and ODC expression also can rise and lack of proper care.Polyamine level increase, along with cell Proliferation accelerate, apoptosis reduce, with
And tumor-infiltrated and metastasis related gene expression raising etc..Therefore, the regulation of polyamines, becomes oncotherapy and drug is ground
An important means in hair.
The starting material of Polyamine Metabolism is ornithine (ornithine), it is arginine in urea cycle (urea
Cycle the reaction product being catalyzed in) by arginase (arginase).ODC is first enzyme of polyamines route of synthesis, catalysis from
Ornithine (ornithine) arrives the reaction of putrescine, and step catalysis reaction is also a rate-limiting step of polyamines route of synthesis.Cause
This, synthesizes ODC inhibitor, inhibits the generation of putrescine, be a currently very popular oncotherapy approach.Simultaneously as sick
Pathogenic microorganism is also required to normal polyamine level, and ODC inhibitor also becomes for pathogenic microorganism (as led to African typanosomiasis nagana
Trypanosoma bocagei) important target.
Currently, the inhibitor DFMO (alpha-difluoromethyl ornithine) of ODC has been used for clinic, assists the chemotherapy of cancer.
But it is weaker with the binding ability of ODC, and activity is very high, and due to being the suicide inhibitor for forming covalent bond with ODC, poison
Side effect is very big.Therefore, it is highly desirable to develop the ODC new inhibitor with more preferable effect.
Summary of the invention
The purpose of the present invention is provide a kind of for the novel small of ODC by computer assisted high-flux medicaments sifting
Molecule inhibitor is applied to the inhibitor of ornithine decarboxylase, may can be used for preparing treatment tumour, pathogenic microorganism
The drug of infection.Specifically:
A kind of micromolecular inhibitor containing benzodioxole, the structural formula of the micromolecular inhibitor are as follows:
Answering on ornithine decarboxylase (ODC) is being inhibited using above-mentioned micromolecular inhibitor containing benzodioxole
With.
Using application of the micromolecular inhibitor containing benzodioxole in preparation tumor.
Above-mentioned micromolecular inhibitor containing benzodioxole is used to inhibit the side of ornithine decarboxylase (ODC)
Method includes the following steps:
1) building of ODC prokaryotic expression plasmid
The gene order of ODC is inserted into pET28a plasmid, is constructed by BamH I and Xho I restriction enzyme site
PET28a-hODC plasmid, is verified through DNA sequencing；
2) expression of ODC albumen
The pET28a-hODC plasmid that step 1) constructs is passed through into CaCl2Method is transformed into e. coli strain bl21, is led to
It crosses kanamycins to be screened, the bacterial strain that then will be grown on Luria-Bertani (LB) culture plate containing kanamycin,
It is seeded in LB liquid medium containing kanamycin, cultivates in 37 DEG C, 250rpm to logarithmic growth phase, then add isopropyl
Thiogalactoside (IPTG) to 0.5mM, 28 DEG C inducing expression 4 hours, finally, bacterium is collected by centrifugation；
3) purifying of ODC albumen
The bacterium that will be collected in step 2), is suspended again with lysate, then carries out cell cracking by ultrasonic method, then
By lysate in 4 DEG C, 12000 turns/min, after centrifugation, retain supernatant；Supernatant is finally utilized into Ni-NTA His label protein knot
It closes filler to be combined and purify, obtains source of people ODC albumen, the elution buffer of ODC albumen is 50mM trihydroxy methyl amino first
Alkane (Tris)/HCl, pH 8.0,300mM NaCl, 1mM DTT, 100mM imidazoles (imidazole)；
4) Activity determination of ODC albumen
In EP pipe, the ODC albumen of 400uL substrate reactions mixture, 50ug is added, EP pipe is placed on 37 DEG C after mixing
30min in water-bath；The trichloroacetic acid that 400uL 10% is added terminates reaction, and room temperature is centrifuged 5000rpm, 5min, then takes out
100uL supernatant is mixed with the NaOH solution of 200uL 4mol/L, and 400uL n-amyl alcohol is added, is sufficiently mixed, 2000rpm centrifugation
5min, then upper liquid 200uL is transferred in new EP pipe, it is equal that the sodium tetraborate mixing that 200uL0.1mol/LpH is 8.0 is added
Even, addition 200uL10mmol/L trinitrobenzene sulfonic acid mixes abundant, addition 400uL DMSO, is sufficiently mixed 1min, 3000rpm
It is centrifuged 5min；Upper liquid is finally taken out into 96 orifice plates, with the light absorption value at microplate reader detection 426nm, obtains not enzyme OD
Value
5) detection of the inhibitor to the inhibitory activity of ODC albumen
Ornithine decarboxylation is added after 400uL substrate reactions mixture is added in the method according to step 4) immediately
The micromolecular inhibitor of enzyme, the same step 4) of subsequent operation；
It is calculated by the following formula out ODC inhibiting rate:
Compare it is poor=plus micromolecular inhibitor control group mean OD value-do not add micromolecular inhibitor control group mean OD value,
The inhibitor that wherein control group is added in step 5) is DFMO inhibitor；
Test it is poor=plus micromolecular inhibitor experimental group mean OD value-micromolecular inhibitor experimental group group is not added to be averaged OD
Value；
ODC inhibiting rate=[(control difference-experiment is poor)/control is poor] × 100%.
Lysate described in the step 3) be 50mM trishydroxymethylaminomethane (Tris)/HCl, pH 8.0,
The mixed liquor of 300mM NaCl, 1mM DTT, 1mM PMSF, 5mM imidazoles (imidazole).
Substrate reactions mixture in the step 4) is in the phosphate buffer (PBS) that 150mM pH is 7.1
Dissolve 17.57ul beta -mercaptoethanol, 55.84mg 1.5mM EDTA disalt sodium, 75nM phosphopyridoxal pyridoxal phosphate (PLP) liquid storage, 2mM bird
Propylhomoserin hydrochloride.
The ornithine decarboxylase be source of people ornithine decarboxylase, non-source of people ornithine decarboxylase or with source of people ornithine
The putrescine substrate of decarboxylase and the protein of phosphopyridoxal pyridoxal phosphate binding site very high homology.
According to above scheme, first with Pocket pharmaceutical grade protein pocket analysis software, with the crystal structure of source of people ODC
Based on, its putrescine substrate and PLP ligand binding pocket are analyzed, and generate the theoretical mould of the protein pocket
Type.Then, 190,000 small molecules in SPECS small-molecule drug library are docked to above-mentioned using protein docking procedure DOCK
In protein bag model, and filter out at least containing 15 non-hydrogen heavy atoms, at least formed 2 hydrogen bonds, at least one it is hydrophobic in
The heart, docking marking are no more than -10 small molecule.Protein-small molecule docking procedure is further utilized to above-mentioned small molecule again
Autodock is successively docked to again in above-mentioned protein pocket, is carried out further docking and is calculated, finally picks out docking
Marking is lower than -7 small molecule.
The present invention also provides a kind of composition, contain a effective amount of provided by the invention small to inhibition ornithine decarboxylase
Molecule inhibitor or its analog and pharmaceutically useful carrier.It is preferred that the composition is pharmaceutical composition, contain to controlling
Treat a effective amount of micromolecular inhibitor provided by the invention or its analog and pharmaceutically useful carrier.The more preferable drug
Composition is the pharmaceutical composition for treating or preventing the disease of inhibition generation response of ornithine decarboxylase (ODC), wherein bird ammonia
Acid decarboxylase (ODC) preferably humanized's ornithine decarboxylase (ODC)；It further include containing the inhibition for preventing and treating ornithine decarboxylase
The micromolecular inhibitor provided by the invention or its analog of the condition effective amount of generation response and pharmaceutically useful carrier.
Micromolecular inhibitor of the invention or its analog and above-mentioned composition can be used for inhibiting ornithine decarboxylase
(ODC), preferred humanized's ornithine decarboxylase (ODC), wherein described inhibit to be therapeutic purposes or non-treatment purpose.It is preferred that this hair
Bright micromolecular inhibitor or its analog are used to prepare the drug for inhibiting ornithine decarboxylase (ODC), preferably humanized bird ammonia
Acid decarboxylase (ODC).Therefore the present invention also provides the methods for inhibiting ornithine decarboxylase activity, and this method is including being needed to press down
The object of ornithine decarboxylase (ODC) processed applies a effective amount of micromolecular inhibitor of the invention or its analog or above-mentioned group
Object is closed, wherein described inhibit to be therapeutic purposes or non-treatment purpose.
The present invention also provides treatments to the method for the disease of the inhibition generation response of ornithine decarboxylase, and this method includes
To the individual application prevention for needing this treatment or prevention or treat upper a effective amount of inhibition ornithine decarboxylase (preferably humanized
Ornithine decarboxylase (ODC)) micromolecular inhibitor of the invention or its analog or above-mentioned composition.
Micromolecular inhibitor of the invention or its analog can be used for preparing treatment and generate to the inhibition of ornithine decarboxylase
The drug or pharmaceutical composition of the disease of response, wherein ornithine decarboxylase (ODC) or its analog inhibit ornithine decarboxylase
Activity, the disease includes tumour or cause pathogeny imcrobe infection, preferably above-mentioned tumour.Protozoan infection refers to ornithine decarboxylation
The inhibition of enzyme generates the tumor disease or cause pathogeny imcrobe infection disease of response.
Detailed description of the invention
Fig. 1 is inhibitory effect of the micromolecular inhibitor to source of people ornithine decarboxylase of embodiment 1.
Fig. 2 is fragmentation effect of the embodiment 1 using mtt assay detection inhibitor to tumour cell.
Specific embodiment
Embodiment 1
Related micromolecular inhibitor are as follows: N'- ({ 6- nitro -1,3- benzodioxole -5- base } methylene
Base) spiral shell [2.3] hexane -1- carbohydrazide, structural formula is as shown in the figure.
Activity test method is as follows:
1. the building of source of people ODC prokaryotic expression plasmid
The gene order of source of people ODC is inserted into pET28a plasmid by BamH I and Xho I restriction enzyme site, is constructed
PET28a-hODC plasmid out is verified through DNA sequencing.
The gene order of source of people ODC:
atgaacaactttggtaatgaagagtttgactgccacttcctcgatgaaggttttactgccaaggacat
tctggaccagaaaattaatgaagtttcttcttctgatgataaggatgccttctatgtggcagacctgggagacatt
ctaaagaaacatctgaggtggttaaaagctctccctcgtgtcacccccttttatgcagtcaaatgtaatgatagca
aagccatcgtgaagacccttgctgctaccgggacaggatttgactgtgctagcaagactgaaatacagttggtgca
gagtctgggggtgcctccagagaggattatctatgcaaatccttgtaaacaagtatctcaaattaagtatgctgct
aataatggagtccagatgatgacttttgatagtgaagttgagttgatgaaagttgccagagcacatcccaaagcaa
agttggttttgcggattgccactgatgattccaaagcagtctgtcgtctcagtgtgaaattcggtgccacgctcag
aaccagcaggctccttttggaacgggcgaaagagctaaatatcgatgttgttggtgtcagcttccatgtaggaagc
ggctgtaccgatcctgagaccttcgtgcaggcaatctctgatgcccgctgtgtttttgacatgggggctgaggttg
gtttcagcatgtatctgcttgatattggcggtggctttcctggatctgaggatgtgaaacttaaatttgaagagat
caccggcgtaatcaacccagcgttggacaaatactttccgtcagactctggagtgagaatcatagctgagcccggc
agatactatgttgcatcagctttcacgcttgcagttaatatcattgccaagaaaattgtattaaaggaacagacgg
gctctgatgacgaagatgagtcgagtgagcagacctttatgtattatgtgaatgatggcgtctatggatcatttaa
ttgcatactctatgaccacgcacatgtaaagccccttctgcaaaagagacctaaaccagatgagaagtattattca
tccagcatatggggaccaacatgtgatggcctcgatcggattgttgagcgctgtgacctgcctgaaatgcatgtgg
gtgattggatgctctttgaaaacatgggcgcttacactgttgctgctgcctctacgttcaatggcttccagaggcc
gacgatctactatgtgatgtcagggcctgcgtggcaactcatgcagcaattccagaaccccgacttcccacccgaa
gtagaggaacaggatgccagcaccctgcctgtgtcttgtgcctgggagagtgggatgaaacgccacagagcagcct
gtgcttcggctagtattaatgtgtag
The above-mentioned source of people ODC sequence that we use, with database (http://www.ncbi.nlm.nih.gov/
Nuccore/NM_002539.1 the sequence in) has a base variation, and (C of box label is T in database sequence, corresponding
Amino acid cysteine is become from arginine), but do not influence its activity.
2. the expression of source of people ODC albumen
Above-mentioned pET28a-hODC plasmid is passed through into CaCl2Method is transformed into e. coli strain bl21, and by card, that is mould
Element is screened.The bacterial strain that will be grown on Luria-Bertani (LB) culture plate containing kanamycin, is seeded to containing card
In the LB liquid medium of that mycin, 37 DEG C, 250rpm cultivate to logarithmic growth phase, then add IPTG (isopropylthio half
Lactoside) to 0.5mM, 28 DEG C inducing expression 4 hours.Finally, bacterium is collected by centrifugation.
3. the purifying of source of people ODC albumen
By the bacterium of above-mentioned collection, with lysate (50mM Tris/HCl, pH 8.0,300mM NaCl, 1mM DTT, 1mM
PMSF, 5mM imidazole.) it suspends again, cell cracking is then carried out by ultrasonic method.Then by lysate 4 DEG C,
After 12000 turns of centrifugations, retain supernatant.Supernatant is combined and is purified using Ni-NTA His label protein combination filler, people
The elution buffer of source ODC albumen is 50mM Tris/HCl, pH 8.0,300mM NaCl, 1mM DTT, 100mM
imidazole。
4. the Activity determination of source of people ODC albumen
In the EP pipe of 1.5mL, (the dissolution in 150mM PBS (pH 7.1) of 400uL substrate reactions mixture is added
17.57ul beta -mercaptoethanol, 55.84mg 1.5mM EDTA disalt sodium, 75nM PLP liquid storage, 2mM ornithine hydrochloride).So
Afterwards, the ODC albumen of 50ug is added.EP pipe is placed on 30min in 37 DEG C of water-baths after mixing.Then it is added the three of 400uL 10%
Monoxone terminates reaction.Room temperature is centrifuged 5000rpm, 5min.100uL supernatant is taken out, it is mixed with the NaOH solution of 200uL 4mol/L
It closes, adds 400uL n-amyl alcohol, be sufficiently mixed.2000rpm is centrifuged 5min, upper liquid 200uL is transferred in new EP pipe,
200uL sodium tetraborate (0.1mol/L, pH8.0) is added to be sufficiently mixed.Then 200uL trinitrobenzene sulfonic acid (10mmol/ is added
L), it is sufficiently mixed.400uL DMSO is added, 1min is sufficiently mixed.3000rpm is centrifuged 5min.Upper liquid is taken out to 96 orifice plates
In, with the light absorption value at microplate reader detection 426nm.
5. the detection of the inhibitory activity of inhibitor on human source ODC albumen
In above-mentioned Activity determination step, after 400uL substrate reactions mixture is added, small-molecule drug is added immediately and mixes
It closes.Subsequent step is the same.
It is calculated by the following formula out ODC inhibiting rate
Compare poor=enzyme not enzyme control group mean OD value of control group mean OD value-
Test poor=enzyme not enzyme experimental group group mean OD value of experimental group mean OD value-
Inhibiting rate=(control difference-experiment poor)/control is poor × and 100%
The inhibitory effect of the micromolecular inhibitor obtained according to the method described above is as shown in Figure 1, as can be seen from the figure exist
50% or more inhibit concentration when, which can inhibit the activity of ODC, and rejection ability is suitable with the DFMO of 2.5mM.
6. using mtt assay detection inhibitor to the fragmentation effect of tumour cell
Using mtt assay detection inhibitor to the fragmentation effect of tumour cell
By A549 cell inoculation into 96 porocyte culture plates, 2000 cells/wells of inoculum density, culture medium RPMI-
1640,10% calf serum, 1% chain/penicillin, the hole volume 100uL/.After being cultivated 24 hours in 37 DEG C of cell incubators, add
Enter the 100uL diluted small-molecule drug of RPMI-1640 or culture medium.After continuing culture 60 hours, culture medium is sucked, is added
After 100uL MTT (final concentration 250ug/mL), continue culture 4 hours.Then culture solution is sucked, 150uL DMSO is added, is shaking
Low-speed oscillation 15min on bed, dissolves crystal sufficiently.Finally, reading the light absorption value at 490nm using microplate reader.
The mtt assay detection inhibitor obtained according to the method described above is to the fragmentation effect of tumour cell as shown in Fig. 2, from figure
It can be seen that the drug is able to suppress and killing tumor cell, but presses down below 50% in the concentration inhibited 50% or more
Ability is very weak when the concentration of system.
Claims (4)
1. a kind of micromolecular inhibitor containing benzodioxole, which is characterized in that the structural formula of the micromolecular inhibitor
Are as follows:
2. micromolecular inhibitor containing benzodioxole described in claim 1 is inhibiting ornithine decarboxylase (ODC)
On application.
3. micromolecular inhibitor containing benzodioxole described in claim 1 answering in preparation tumor
With.
4. micromolecular inhibitor containing benzodioxole described in claim 1 treats cause pathogeny imcrobe infection in preparation
Application in drug.
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| WO2006121684A2 (en) * | 2005-05-06 | 2006-11-16 | Boehringer Ingelheim International, Gmbh | Acyl hydrazones for treating cardiovascular diseases |
| WO2017217796A1 (en) * | 2016-06-15 | 2017-12-21 | 국립암센터 | Rhoa inhibitor and use thereof |
| CN107827776A (en) * | 2017-11-10 | 2018-03-23 | 上海应用技术大学 | Acylhydrazone, preparation method and applications with antitumor activity |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006121684A2 (en) * | 2005-05-06 | 2006-11-16 | Boehringer Ingelheim International, Gmbh | Acyl hydrazones for treating cardiovascular diseases |
| WO2017217796A1 (en) * | 2016-06-15 | 2017-12-21 | 국립암센터 | Rhoa inhibitor and use thereof |
| CN107827776A (en) * | 2017-11-10 | 2018-03-23 | 上海应用技术大学 | Acylhydrazone, preparation method and applications with antitumor activity |
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