CN107474117A - A kind of antitumor cyclic peptide and preparation method thereof and medical applications - Google Patents
A kind of antitumor cyclic peptide and preparation method thereof and medical applications Download PDFInfo
- Publication number
- CN107474117A CN107474117A CN201710667566.3A CN201710667566A CN107474117A CN 107474117 A CN107474117 A CN 107474117A CN 201710667566 A CN201710667566 A CN 201710667566A CN 107474117 A CN107474117 A CN 107474117A
- Authority
- CN
- China
- Prior art keywords
- cyclic peptide
- cell
- antitumor
- antitumor cyclic
- preparation
- 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.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/64—Cyclic peptides containing only normal peptide links
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
A kind of antitumor cyclic peptide and preparation method thereof and medical applications, belong to field of biological pharmacy.Antitumor cyclic peptide, its amino acid sequence are
Description
Technical field
The invention belongs to field of biological pharmacy, and in particular to a kind of antitumor cyclic peptide and preparation method thereof and medical applications.
Background technology
In recent years, using the polypeptide drugs of modern biotechnology synthesis as one of focus of medicament research and development, it is because suitable
Wide, safe and evident in efficacy, prevention, diagnosis and the treatment applied to related significant diseases such as tumours at present, tool should be demonstrate,proved
There is wide DEVELOPMENT PROSPECT.
As polypeptide drugs turn into one of indispensable key areas of research and development modern medicines, its development is also more and more faster.
Having had more than 300 kinds of polypeptide drugs enters clinical experimental stage at present, has more than 600 kinds of polypeptide drugs just in preclinical test
In, and 60 kinds of polypeptide drugs listings are had more than, including for treating the polypeptide drugs of tumour, be such as used to treat resectable marrow
The Mepect of knurl, the Istodax for treating skin T cell lymphoma and for treating CD30 positive Hodgkin lymphomas
Adcetris etc..Polypeptide drugs molecular structure is small, easy transformation, is easily-synthesized, and the waste discharged in production process is few, belongs to green
Pharmacy, therefore polypeptide drugs are one of 21 century most promising medicines.
The content of the invention
The invention provides a kind of antitumor cyclic peptide, the amino acid sequence of the antitumor cyclic peptide is
Two of which Cys forms disulfide bond.
The molecular weight of the antitumor cyclic peptide is 1442.71, can be purified by reversed-phased high performace liquid chromatographic, and
Purifying can make its purity reach 96.78% by this way.
The invention provides a kind of anti-tumor compositions, the anti-tumor compositions include anti-tumor cyclic of the present invention
Peptide.
The present invention further discloses the preparation method of the antitumor cyclic peptide, the present invention is made using solid-phase synthesis
The standby antitumor cyclic peptide.The solid-phase synthesis can be reached by following measures：
Fmoc schemes are taken, CTC macromolecule resins is chosen, is the Fmoc-Gly- of c-terminus by the anti-tumor cyclic PEPC end
OH is connected in the form of covalent bond on CTC resins, takes off Fmoc blocking groups, with the ammonia for the Gly being incorporated on macromolecule resin
Base as synthesis starting point, according to the antitumor cyclic peptide from C-terminal be c-terminus to the N-terminal i.e. amino acid sequence of aminoterminal
CGHFKFGYLRCG order, carry out condensation reaction successively respectively and form peptide bond, take off Fmoc protections, condensation forms peptide bond, constantly
Repetitive cycling the step, until sequence, all coupling is complete, takes off Fmoc protections；Afterwards, polypeptide is obtained with methanol cutting resin
Crude product, it is further purified using reversed-phased high performace liquid chromatographic, two Cys is formed disulfide bond cyclization by processing；Again with anti-
Phase high performance liquid chromatography purifies to obtain the target cyclic peptide of high purity 96.78%.
The invention provides application of the antitumor cyclic peptide in antineoplastic is prepared.The tumour be selected from liver cancer,
Cervical carcinoma and lung cancer.The antitumor cyclic peptide is in 5-500 μM of concentration range to HepG-2 cell and cervical cancer cell
Hela proliferation inhibition rate is respectively 2.6%-100% and 3.9%-99.9%, and the antitumor cyclic peptide is in 9-900 μM of concentration model
It is 16.5%-99.3%, its IC to enclose the interior proliferation inhibition rate to lung cell A54950Respectively 36.90 μM of value, 61.41 μM,
39.87μM。
The invention provides the antitumor cyclic peptide to prepare the application in being used to promote apoptosis of tumor cells medicine.
In one embodiment, the tumour cell is selected from liver cancer cells, cervical cancer cell and lung carcinoma cell.
In one embodiment, the tumour cell is lung carcinoma cell.It is right when the anti-tumor cyclic peptide concentration is 40 μM
The rush apoptosis rate of lung cell A549 is 40.49%.
Compared with prior art, the present invention has the following advantages that and technique effect：It is anti-swollen that the present invention has synthesized this first
Knurl cyclic peptide, and use CCK-8 methods to determine the antitumor activity of the cyclic peptide, the antitumor cyclic peptide, which has, significantly to be suppressed
The ability of tumor cell proliferation.In addition, the present inventor also have detected rush of the cyclic peptide to tumour cell using Flow Cytometry
Apoptotic effect, as a result find, the antitumor cyclic peptide has significant apoptosis-promoting effect to tumour cell.
Brief description of the drawings
The antitumor cyclic peptide of Fig. 1HPLC figure.
The antitumor cyclic peptide of Fig. 2MS figure.
The antitumor cyclic peptide of the invention that Fig. 3 concentration is 40 μM detects figure to the rush Apoptosis of lung cell A549.
Embodiment
The embodiment of the present invention illustrates with the following Examples, but the present invention is not limited to following examples.
The synthesis in solid state of 1 antitumor cyclic peptide of embodiment
During the synthesis in solid state of antitumor cyclic peptide of the present invention, using Fmoc schemes, CTC resins are chosen, are entered
Row processing makes resin expose avtive spot, and Fmoc-Gly-OH is connected on CTC resins in the form of covalent bond, washed with DMF
Resin.Add volumetric concentration and take off Fmoc blocking groups for 25% piperidines/DMF solution.Utilize the synthesis in solid state process of standard
(SPPS) HBTU, N-methylmorpholine and Fmoc-Cys (Trt)-OH, are added, makes Fmoc-Cys (Trt)-OH with being connected to
Gly on CTC resins occurs dehydration condensation and linked together by peptide bond, and whether ninhydrin detection reaction is complete.Again plus
Enter volumetric concentration and take off Fmoc blocking groups for 25% piperidines/DMF solution；Using above-mentioned steps, according to amino acid sequence
CGHFKFGYLRCG's is sequentially connected with up, and until sequence, all coupling is complete, takes off Fmoc protections.Resin is washed with methanol
Wash to be cut.Transfer a resin into cutting pipe, addition includes TFA, H in the prior art2O, phenol, EDT, thioanisole etc. one
The mixed solution of kind solution or several solns, ether precipitation, has cut resin and has obtained polypeptide crude product.Utilize RPLC
Method is further purified, and is then dissolved in pure water, by 1% I2/ MeOH solution (1g I2It is dissolved in 100ml MeOH) slowly drip
It is added in solution, two Cys is formed disulfide bond cyclization.The aqueous solution is extracted with dichloromethane, uses reversed-phased high performace liquid chromatographic
Purifying obtains the antitumor cyclic peptide that purity is 96.78%.
Embodiment 2 determines cyclic peptide to HepG-2 cell, cervical cancer cell Hela and lung carcinoma cell using CCK-8 methods
The inhibitory action of A549 propagation
By HepG-2 cell cell suspension (1.5 × 104Individual/mL), cervical cancer cell Hela cell suspensions (2 × 104
Individual/mL) and lung cell A549 cell suspension (2 × 104Individual/mL) according to every μ L of hole 200 volume it is separately added into 96 orifice plates
It is interior, in 37 DEG C of constant temperature CO2Incubator culture.Cell attachment grows after 24h, suctions out old nutrient solution, antitumor cyclic peptide is pressed
Prepare as 5-500 μM and 9-900 μM of decoction, be separately added into corresponding 96 orifice plate respectively according to the concentration of Tables 1 and 2, and
Negative control and blank control are set, in 37 DEG C of constant temperature CO2Incubator is incubated.After 48h, decoction is suctioned out, adds what is prepared
CCK-8 solution, continue at 37 DEG C of constant temperature CO2Incubator culture.After 2h, its extinction at 450nm wavelength is surveyed on ELIASA
Angle value.As shown in Table 1, in the presence of 5-500 μM of concentration, the extracorporeal inhibiting rate to HepG-2 cell is antitumor cyclic peptide
2.6%-100%, its IC50It is worth for 36.90 μM, the inhibiting rate to cervical cancer cell Hela is 3.9%-99.9%, its IC50Value
For 61.41 μM.As shown in Table 2, in the presence of 9-900 μM of concentration, the inhibiting rate to lung cell A549 is antitumor cyclic peptide
16.5%-99.3%, its IC50It is worth for 39.87 μM.
The inhibitory action that 1 antitumor cyclic peptide of table is bred to tumour cell HepG-2 and Hela cell
The inhibitory action that 2 antitumor cyclic peptide of table is bred to tumour cell A549
Apoptosis-promoting effect of the embodiment 3 using flow cytometry detection cyclic peptide to lung cell A549
Lung cell A549 is chosen as research object, detects apoptosis-promoting effect of this cyclic peptide to tumour cell.By logarithm
The lung cell A549 cell suspension (1.75 × 10 in growth period4Individual/mL) added according to every hole 2mL volume in 6 orifice plates, in 37
DEG C constant temperature CO2Incubator culture.Cell attachment grows after 24h, adds final concentration of 40 μM of antitumor cyclic peptide decoction, sets not
Dosing only adds the negative control of culture medium and cell, and the mono- positive controls contaminated of dosing FITC/PI, in 37 DEG C of constant temperature CO2Culture
Case culture.It is standby in handled cell culture fluid to centrifuge tube after 48h, with the trypsin digestion cell without EDTA, before addition
The nutrient solution that face is collected carefully blows and beats cell.Centrifugation cell, cell is washed with the PBS of 4 DEG C of precoolings, centrifugation is thin again
Born of the same parents.
Cell is resuspended with the combination buffer for preparing concentration, it is 1-5 × 10 to adjust its concentration6/mL.Take that 100 μ L's is thin
Born of the same parents' suspension adds Annexin V/FITC and mixed after room temperature lucifuge incubation 5min, it is molten to add propidium iodide in 5mL streaming pipes
Liquid (PI) and PBS, carry out flow cytometer detection on flow cytometer.
Testing result is as shown in figure 3, it will be apparent from this figure that this cyclic peptide has apoptosis-promoting effect to lung cell A549.
Final result shows that this cyclic peptide is 40.49% to the rush apoptosis rate of lung cell A549 under 40 μM of concentration.
Similarly, with identical method of testing, the results showed that the antitumor cyclic peptide is for HepG-2 cell and palace
Neck cancer cell Hela, which also has, promotees cells apoptosis.
Claims (8)
1. a kind of antitumor cyclic peptide, it is characterised in that the amino acid sequence of the antitumor cyclic peptide is
Two of which Cys forms disulfide bond.
2. a kind of anti-tumor compositions, it is characterised in that the anti-tumor compositions include the anti-tumor cyclic described in claim 1
Peptide.
3. a kind of preparation method for preparing antitumor cyclic peptide described in claim 1, it is characterised in that made using solid-phase synthesis
The standby antitumor cyclic peptide.
4. the antitumor cyclic peptide described in claim 1 is as the application for preparing antineoplastic.
5. the application described in claim 4, it is characterised in that the tumour is selected from liver cancer, cervical carcinoma and lung cancer.
6. the antitumor cyclic peptide described in claim 1 is preparing the application in being used to promote apoptosis of tumor cells medicine.
7. application according to claim 6, it is characterised in that the tumour cell is selected from liver cancer cells, cervical cancer cell
And lung carcinoma cell.
8. application according to claim 7, it is characterised in that the tumour cell is lung carcinoma cell.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710667566.3A CN107474117B (en) | 2017-08-07 | 2017-08-07 | Antitumor cyclopeptide and preparation method and medical application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710667566.3A CN107474117B (en) | 2017-08-07 | 2017-08-07 | Antitumor cyclopeptide and preparation method and medical application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107474117A true CN107474117A (en) | 2017-12-15 |
| CN107474117B CN107474117B (en) | 2020-08-07 |
Family
ID=60598763
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710667566.3A Active CN107474117B (en) | 2017-08-07 | 2017-08-07 | Antitumor cyclopeptide and preparation method and medical application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107474117B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109503701A (en) * | 2018-07-20 | 2019-03-22 | 北京工业大学 | A kind of cyclic peptide and its application in preparation of anti-tumor drugs |
| CN112480213A (en) * | 2020-11-27 | 2021-03-12 | 常州大学 | Amphiphilic anti-tumor polypeptide and application thereof |
| CN114478701A (en) * | 2022-03-18 | 2022-05-13 | 广西民族大学 | Heptapeptide with anti-tumor activity and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110305706A1 (en) * | 2009-02-23 | 2011-12-15 | Scott Thomas Brady | Compositions and Methods for Treating a Disease Mediated by Soluble Oligomeric Amyloid Beta |
| CN103665106A (en) * | 2012-09-20 | 2014-03-26 | 天津嘉宏科技有限公司 | Antineoplastic polypeptide and preparation method and applications thereof |
| CN106397544A (en) * | 2016-09-06 | 2017-02-15 | 中国科学院南海海洋研究所 | Application of cyclic peptide compounds in preparation of antitumor drug |
-
2017
- 2017-08-07 CN CN201710667566.3A patent/CN107474117B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110305706A1 (en) * | 2009-02-23 | 2011-12-15 | Scott Thomas Brady | Compositions and Methods for Treating a Disease Mediated by Soluble Oligomeric Amyloid Beta |
| CN103665106A (en) * | 2012-09-20 | 2014-03-26 | 天津嘉宏科技有限公司 | Antineoplastic polypeptide and preparation method and applications thereof |
| CN106397544A (en) * | 2016-09-06 | 2017-02-15 | 中国科学院南海海洋研究所 | Application of cyclic peptide compounds in preparation of antitumor drug |
Non-Patent Citations (4)
| Title |
|---|
| BENOIT BESTGEN等: "In Search of Small Molecule Inhibitors Targeting the Flexible CK2 Subunit Interface", 《PHARMACEUTICALS》 * |
| LAUDET等: "Structure-based design of small peptide inhibitors of protein kinase CK2 subunit interaction", 《BIOCHEM.J.》 * |
| SHAN TANG等: "Structure-based Discovery of Novel CK2α-Binding Cyclic Peptides with Anti-cancer Activity", 《MOLECULAR INFORMATICS》 * |
| 周玥: "蛋白激酶CK2抑制剂作用机理的分子模拟研究及其靶向设计", 《中国博士学位论文全文数据库（电子期刊）医药卫生科技辑》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109503701A (en) * | 2018-07-20 | 2019-03-22 | 北京工业大学 | A kind of cyclic peptide and its application in preparation of anti-tumor drugs |
| CN109503701B (en) * | 2018-07-20 | 2022-03-15 | 北京工业大学 | Cyclopeptide and application thereof in preparation of antitumor drugs |
| CN112480213A (en) * | 2020-11-27 | 2021-03-12 | 常州大学 | Amphiphilic anti-tumor polypeptide and application thereof |
| CN114478701A (en) * | 2022-03-18 | 2022-05-13 | 广西民族大学 | Heptapeptide with anti-tumor activity and preparation method and application thereof |
| CN114478701B (en) * | 2022-03-18 | 2024-04-19 | 广西民族大学 | Heptapeptide with anti-tumor activity, and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107474117B (en) | 2020-08-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100509857C (en) | High substitution degree carboxymethyl indianbread polysaccharide, preparation method and uses thereof | |
| CN107474117A (en) | A kind of antitumor cyclic peptide and preparation method thereof and medical applications | |
| CN103694320B (en) | A kind of preparation method of that peptide of pulika | |
| CN104630318B (en) | A kind of preparation method of small water turtle antineoplastic polypeptide | |
| CN101143894A (en) | Highly effective polypeptide for inhibiting angiogenesis, physical chemistry modifying method and application thereof | |
| CN104232718B (en) | The preparation method and application of dendrobium candidum antineoplastic polypeptide | |
| CN103204907B (en) | Raja porosa cartilage polypeptide angiogenesis inhibitory factor, and preparation method and application thereof | |
| CN104106763B (en) | Polygonatum polysaccharide has the application in the auxiliary functional food suppressing colorectal carcinoma effect in preparation | |
| CN104530199A (en) | Antitumor polypeptide, and preparation method and application thereof | |
| CN107849551A (en) | New trypsase is the same as work(iso series and its purposes | |
| CN112386707A (en) | Tumor-targeted polypeptide drug conjugate and preparation method thereof | |
| CN101041692A (en) | Scorpion analgesic antibacterial active peptide and preparation method thereof | |
| CN1723955A (en) | Extractive of rhizome belamcandae, prepn. method and use thereof | |
| CN107337715A (en) | A kind of antitumor cyclic peptide and its preparation and application | |
| CN105237624B (en) | A kind of heptapeptide EMLQPPL and its application | |
| CN109503701B (en) | Cyclopeptide and application thereof in preparation of antitumor drugs | |
| WO2020108427A1 (en) | NOVEL INTERFERON-α AND PREPARATION METHOD THEREFOR, COMPOSITION AND USE THEREOF | |
| CN106928335A (en) | Her-2 polypeptides, composition and preparation method and application for tumour | |
| WO2007054030A1 (en) | Polyethylene glycol modifications of thymosin alpha-1 | |
| CN111072784B (en) | Macromolecular furin inhibitor and preparation method and application thereof | |
| CN110144015B (en) | Method for synchronously extracting and purifying ganoderan, ganoderma triterpenic acid and amino acid | |
| CN109467597B (en) | Novel interferon and preparation method, composition and application thereof | |
| CN103232543B (en) | Recombinant protein Tumstatin-CD137L4 with Tumstatin activities as well as preparation method and application thereof | |
| CN110551179B (en) | Modified anti-HIV polypeptide and preparation method and application thereof | |
| CN101880327A (en) | Scorpion arialgesic anti-tumoral peptide fusion and acquisition method thereof |
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 |