CN114805493A - Preparation and anti-tumor effect of RGD/KLA integrated lipopeptide - Google Patents
Preparation and anti-tumor effect of RGD/KLA integrated lipopeptide Download PDFInfo
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Abstract
The invention relates to an anticancer lipopeptide C 8 H 15 The preparation of O-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Asp-Gly-Arg and the application thereof in the anti-tumor treatment belong to the field of biological medicine. The anticancer lipopeptides are prepared based on key biological features of membrane-cleavable peptides (positive charge, alpha-helical structure and amphiphilicity). The invention provides a solid-phase synthesis method of the anticancer lipopeptide. The anticancer lipopeptide has the characteristics of low hemolytic activity and good serum stability, can effectively make up the defect of high hemolytic toxicity of most anticancer peptides, and solves the problem of high hemolytic toxicity of the anticancer peptidesThe problem of poor in vivo stability of peptide drugs. In vitro anticancer experiments and in vivo antitumor experiments prove that the anticancer lipopeptide has good antitumor effect and good application prospect.
Description
Technical Field
The invention belongs to the field of biomedicine, and particularly relates to preparation of an artificially synthesized anticancer lipopeptide with good anticancer activity and application of the artificially synthesized anticancer lipopeptide in tumor treatment.
Background
Cancer, also called malignant tumor, is a disease caused by abnormality of cell division and proliferation mechanism, and has become one of the most serious diseases threatening human health worldwide. Currently, surgical therapy, radiation therapy (radiotherapy) and chemotherapy (chemotherapy) are the main treatments for cancer. However, the traditional anticancer drugs have high anticancer efficiency, but have some disadvantages, such as low selectivity, obvious side effects, immunosuppression, nerve and gastrointestinal damage and the like; more importantly, the combination of these anti-cancer drugs is highly likely to cause multidrug resistance (MDR) in tumors. Therefore, new breakthroughs are expected for the drug therapy of tumors. Anticancer peptides (ACPs) are bioactive peptides with antitumor activity, such as RGD peptides and KLA peptides. The special action mechanism of the anti-cancer peptide makes the anti-cancer peptide become a hot spot in the research of biological medicines in recent years, and provides a new direction for the research of novel anti-cancer medicines.
Compared with the traditional chemotherapeutic drugs and protein drugs, the polypeptide drugs play an important role in disease treatment due to the advantages of small relative molecular mass, good stability, high biological activity, specificity and the like. Although ACPs have good anticancer activity, the ACPs still have the disadvantages of low cell membrane permeability, poor stability, susceptibility to hydrolysis by proteases, and the like, which limits further development. Therefore, researchers have focused on modifications and alterations to polypeptide drugs to overcome the above-mentioned deficiencies. For example, the polypeptide is modified by fatty acid, so that the polypeptide is not easy to hydrolyze by protease, the stability is improved, and the modified polypeptide drug molecule is enlarged, so that the filtration of glomeruli can be avoided, the half-life period in vivo is prolonged, and the drug effect is improved. Therefore, the present study integrates RGD and KLA peptides and modifies them with fatty acids in anticipation of developing therapeutic drugs of practical significance to alleviate patient suffering. When the inventor searches and compares the whole sequence amino acid structure of the anticancer lipopeptide of the invention by NCBI protein database, no identical polypeptide is found.
Disclosure of Invention
The present invention aims at synthesizing one kind of anticancer lipopeptide with excellent anticancer activity via solid phase synthesis process and applying the anticancer lipopeptide in antitumor treatment.
In order to realize the purpose of the invention, the following technical scheme is provided:
the amino acid sequence of the anticancer lipopeptide is as follows: c 8 H 15 O-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Asp-Gly-Arg, wherein the technical scheme is as follows:
(1) design of anticancer lipopeptide: according to the cationic and amphiphilic structure characteristics of the anticancer peptide, the RGD peptide and the KLA peptide are integrated, and the N terminal is simultaneously provided with N-caprylic acid (C) 8 H 16 O 2 ) Modification is beneficial to improving the lipid solubility and stability of the medicine, and can promote the absorption of cancer cells to the medicine.
(2) Synthesis of anticancer lipopeptide: the anticancer lipopeptide is prepared by a solid-phase synthesis method, wherein the preparation scheme comprises the steps of firstly removing Fmoc protecting groups from amino acids with Fmoc protecting groups and side chain protecting groups coupled on Resin, then catalyzing the amino acids to be coupled on Wang Resin one by one from C end to N end by using a catalyst, then coupling N-octanoic acid at N end, and finally using a cutting fluid (TFA: H) 2 O: tis 95:2.5:2.5) cutting resin, and removing side chain protecting group to obtain the anticancer lipopeptide. The anticancer lipopeptide has the following structural formula:
(3) in vitro anticancer activity studies of anticancer lipopeptides: testing the toxicity of the anti-cancer lipopeptide on H22 cells by MTT method;
(4) secondary structure studies of anticancer lipopeptides: detecting the secondary structure of the anticancer lipopeptide by Circular Dichroism (CD), wherein pure water simulates a hydrophilic environment, and a Sodium Dodecyl Sulfate (SDS) solution simulates a hydrophobic environment of a cell membrane;
(5) research on membrane rupture activity of anticancer lipopeptides: co-culturing the anticancer lipopeptide and H22 cells for 5min and 30min respectively, staining by PI/Hoechst 33342, and observing the influence of the anticancer lipopeptide on the cell membrane morphology under a microscope;
(6) serum stability and hemolytic studies of anticancer lipopeptides: co-culturing with Fetal Bovine Serum (FBS) and anticancer lipopeptide in advance, and detecting the change of anticancer activity after FBS pretreatment by MTT to evaluate the stability of the FBS; evaluating the hemolytic activity of the mouse erythrocyte by the influence of the anticancer lipopeptide on the morphological change of the mouse erythrocyte;
(7) in vivo antitumor activity studies of anticancer lipopeptides: tumor-implanted mice are selected as experimental models, anti-cancer lipopeptide is injected into the tail vein every other day, the weight and the tumor volume of the mice are measured, and the tumor tissues of the mice are dissected and subjected to tissue section and HE staining to detect the in-vivo anti-tumor activity of the anti-cancer lipopeptide.
Drawings
FIG. 1: the MTT method detects the anticancer activity of the anticancer lipopeptide on H22 cells. Relative cell viability of co-cultured H22 cells with various concentrations of anticancer lipopeptide for 24H.
FIG. 2: CD spectra of anticancer lipopeptides in pure water and SDS solutions.
FIG. 3: membrane rupture activity of anticancer lipopeptides. After the lipopeptide and H22 cells are respectively incubated for 5min (a-b) and 30min (c-d), PI/Hoechst 33342 is stained, and the influence of the anticancer lipopeptide on the cell membrane morphology is observed under a microscope.
FIG. 4: hemolytic activity and serum stability of anticancer lipopeptides. Injecting physiological saline (a) and 2.25mg/mL anticancer lipopeptide (b) into the tail vein of the mouse for 2h, then taking blood, and observing the morphology of red blood cells (400 x) under a microscope; (c) the change of anticancer activity of anticancer lipopeptide on H22 cells was measured by MTT method after incubating anticancer lipopeptide with 10% serum and serum-free PBS for 24H at 37 ℃.
FIG. 5: anti-tumor activity of anti-cancer lipopeptides in vivo. (a) Relative tumor volume change with time after NS and anticancer lipopeptide are injected into the tail vein of the tumor-implanted mice every other day; (b) graph of body weight change over time following tail vein injection of NS and anticancer lipopeptides in tumor-implanted mice every other day; (c) tumor tissue sections (200 x) 11 days after saline injection every other day; (d) tumor tissue sections (200X) 11 days after the next day injection of anti-cancer lipopeptide.
Detailed Description
The following is a description of the present invention for further explanation of its constitution, but the present invention is not to be construed as being limited to the following embodiments.
Example 1: preparation method of anticancer lipopeptide
The synthesis of the anticancer lipopeptide is carried out based on a solid-phase polypeptide synthesis method protected by Fmoc. The sequence of the anticancer lipopeptide is C 8 H 15 O-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Asp-Gly-Arg, and the specific synthetic steps are as follows:
(1) synthesis of Fmoc-Lys (Boc) -Ala-Leu-Lys (Boc) -Asp (OtBu) -Gly-Arg (Pbf) -Wang Resin;
Fmoc-Lys (Boc) -Ala-Leu-Lys (Boc) -Ala-Leu-Lys (Boc) -Asp (OtBu) -Gly-Arg (Pbf) -Wang Resin samples were synthesized by solid phase synthesis; swelling Fmoc-Arg (Pbf) -Wang Resin with DMF for 30 min; detecting ninhydrin with a small amount of resin, and adding 20% piperidine-containing DMF solution to react for 30min to remove Fmoc protecting group if no color change exists; washing the resin with DMF, DCM and DMF sequentially, and swelling the resin with anhydrous DMF for 30 min; after swelling, adding 2 times of equivalent of amino acid and 2.6 times of equivalent of DCC, HOBT and DIEA for reaction for more than 48 h; after dialysis, a small amount of resin is taken for ninhydrin detection, and if no color change exists, the corresponding amino acid is connected; then, sequentially washing the resin with DMF, DCM, and DMF, dialyzing in 95% ethanol with dialysis bag (MW8000-14000) for more than 20 times, each time for 30 min; the above steps were repeated until Fmoc-Lys (Boc) -Ala-Leu-Lys (Boc) -Asp (OtBu) -Gly-Arg (Pbf) -Wang Resin samples were synthesized.
(2)C 8 H 15 Synthesis of O-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Asp-Gly-Arg;
first, Fmoc-Lys (Boc) -Ala-Leu-Lys (Boc) -Asp (OtBu) -Gly-Arg (Pbf) -Wang Resin is subjected to Fmoc group removal in the step (1) to obtain Lys (Boc) -Ala-Leu-Lys (Boc) -Ala-Leu-Lys (Boc) -Asp (OtBu) -Gly-Arg (Pbf) -Wang Resin, and freeze-drying the same; 2.5 times of equivalent of NHS and EDC and 2 times of equivalent of n-octanoic acid are put into anhydrous DMF for activation for 5h, and then the lyophilized resin is added for reaction for 48 h; washing the resin with DMF, DCM, and DMF after reaction, loading the resin into dialysis bag (MW8000-14000), dialyzing for more than 20 times (each dialysis time is 3 times)0 min; after the dialysis is finished, washing the dialysis membrane; the resulting sample was lyophilized and cleavage solution (TFA: H) was added 2 O: tis 95:2.5:2.5) cutting off side chain protecting group and resin to obtain target lipopeptide C 8 H 15 O-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Asp-Gly-Arg。
Example 2: in vitro anti-cancer activity assay for anti-cancer lipopeptides
The in vitro anti-cancer activity of anti-cancer lipopeptides was tested by the MTT method. H22 cells were selected in the logarithmic growth phase, H22 cells were seeded into 96-well plates, and then 5% CO was added 2 The lipopeptide was prepared in different concentrations by using a medium at 37 ℃ in an incubator, and the concentrations were set to 0. mu.g/mL, 20. mu.g/mL, 40. mu.g/mL, 60. mu.g/mL, 100. mu.g/mL, 120. mu.g/mL, 140. mu.g/mL, 160. mu.g/mL, 180. mu.g/mL, and 200. mu.g/mL, respectively. 100 μ L of medium containing the drug was added to each well, and 5 wells were set for each concentration to reduce the effect of error due to experimental contingency. After co-culturing the cells and drug for 24h, 20. mu.L (5 mg. multidot.mL) was added to each well -1 ) The MTT solution was cultured in a cell culture chamber for 4 hours. After 4h, the supernatant was discarded, 150. mu.L of DMSO solution was added to each well, and the 96-well plate was placed on a shaker in the dark at 170 r.min -1 And oscillating for 7min, fully dissolving the crystals in the holes, detecting the OD value of the crystals by using an enzyme-labeling instrument, setting the wavelength of the enzyme-labeling instrument to be 485nm, measuring absorbance to calculate the relative survival rate of cancer cells, and evaluating the in-vitro anticancer activity of the lipopeptide.
Example 3: secondary structure determination of anticancer lipopeptides
The secondary structure of lipopeptides in water and SDS solution was examined using Circular Dichroism (CD). Lipopeptides were dissolved in pure water and 30mM SDS solution, respectively, to form lipopeptide solutions at a concentration of 150. mu.M. The wavelength setting range is 180 nm and 260nm, and the optical path selection is 0.5 mm. The SDS solution environment can simulate the surface environment of cancer cell membranes, so that the lipopeptide can be bonded to the cancer cell membranes to observe the change of secondary structures. The data is converted by the following formula:
θ M ＝θobs×1000/cln
θ M is the average residue ovality (deg.cm) 2 ·dmol -1 ) θ obs is the ellipticity (mdeg) observed at a given wavelength, c is the sample concentration (mM), l is the optical path length (mM), and n is the number of residues of the peptide.
Example 4: research on membrane-breaking activity of anticancer lipopeptide on cancer cells
To study the membrane-breaking activity of anticancer lipopeptides, the anticancer lipopeptides were co-cultured with H22 cells for 5min and 30min, respectively, stained with PI/Hoechst 33342, and the effect of the anticancer lipopeptides on the cell membrane morphology was observed under a microscope. PI is a nuclear staining reagent that stains DNA and releases red fluorescence after intercalating into double-stranded DNA. PI cannot enter a living cell membrane to stain, but can cross a disrupted cell membrane to stain nuclei.
Example 5: serum stability and hemolytic Activity assays for anti-cancer lipopeptides
Whether the lipopeptide has serum stability is determined by measuring the change of killing capacity of the lipopeptide to H22 cells in serum-free environment. Meanwhile, a control group and an experimental group are arranged, wherein the control group is the lipopeptide dissolved by PBS, and the experimental group is the lipopeptide dissolved by PBS containing 10% serum. The two groups were incubated in a 37 ℃ cell culture chamber for 24H, and the relative survival rate of H22 cells was calculated by performing MTT experimental studies.
In the hemolytic activity experiment, two female mice were randomly selected, and a physiological saline solution and 2.25mg/mL (15 mg. kg) -1 ) The lipopeptide is injected into a mouse body by a tail vein injection method, blood is taken by an eyeball blood taking mode after 2 hours, the lipopeptide is added with heparin for anticoagulation and then diluted by normal saline, and the erythrocyte morphology is observed under a fluorescence microscope.
Example 6: in vivo antitumor Activity of anticancer lipopeptides
Establishing an in vivo tumor-bearing mouse model to test the in vivo anti-tumor activity of the lipopeptide. The mice used in the experiment are (24-27 g) female Kunming mice, and are strictly raised according to the 'nursing and use guide of animal experiment'. Will be 1 × 10 6 H22 cells were inoculated subcutaneously into the left forelimb axilla of mice when tumors grew 150-220mm 3 The antitumor activity test is carried out. Mice were randomly assigned to give birthSaline group and lipopeptide group, 5 per group. The 2 groups of mice were administered 4 times at an injection dose of 0.2mL and a concentration of 2.25mg/mL -1 The interval between injections in each group was 1 day. The experiment was for a period of 9 days, and the mice were sacrificed by cervical dislocation. The body weight and tumor volume of mice were recorded before each injection and before final dissection, tumors of each group of mice were dissected and then fixed by soaking in 4% paraformaldehyde solution for more than 24h, and finally tumor tissue sections were sectioned and HE stained.
The anticancer lipopeptide prepared by the invention has good biomedical performance:
(1) the anticancer lipopeptide has excellent in vitro anticancer activity
The in vitro anticancer activity of the anticancer lipopeptide is detected by MTT method. As shown in FIG. 1, the relative cell survival rate of H22 cells gradually decreased with the increase of lipopeptide concentration, and was 45.93% at 160. mu.g/mL, indicating that lipopeptide has strong in vitro anticancer effect and concentration dependence on cancer cell killing.
(2) The anticancer lipopeptide has alpha helical structure in hydrophobic cell membrane environment
The secondary structure of the anticancer lipopeptides in water and SDS solutions was examined by circular dichroism. As shown in FIG. 2, the dotted line represents an aqueous solution simulating a hydrophilic environment and the solid line represents an SDS solution simulating a hydrophobic environment of a cell membrane, and it can be seen that the secondary structure of lipopeptides in the environments of water and an SDS solution is greatly different. Wherein a typical alpha helix structure characteristic absorption peak can be seen in the SDS solution, a positive peak is obvious at 195-200 nm, and negative peaks are obvious at 208nm and 222nm, which shows that the alpha helix structure appears after the lipopeptide is dissolved in the SDS solution, but the phenomenon does not appear after the lipopeptide is dissolved in water. Therefore, it is presumed that when the anticancer lipopeptide encounters a cell membrane, it is inserted into the cell membrane in an α -helical structure, and membrane cleavage is performed.
(3) The anticancer lipopeptide has strong membrane penetrating capacity
PI is a nuclear staining reagent for DNA, which releases red fluorescence after intercalation into double-stranded DNA, cannot penetrate living cell membranes to stain, but can cross broken cell membranes to stain nuclei. As can be seen in FIG. 3a, the red fluorescence is very low, indicating that at 5min the H22 cell membrane is still relatively intact. As can be seen from FIG. 3c, the red fluorescence is significantly increased compared to FIG. 3a, indicating that at 30min, the lipopeptide is able to perform a membrane-penetrating action, and thus the H22 cell membrane is changed, and the permeability is significantly increased. The fluorescent dye Hoechst 33342 itself can slightly enter normal cell membranes, staining them with a low blue color, so it can be observed from fig. 3b that at 5min blue fluorescence has appeared due to the entrance of Hoechst 33342. From FIG. 3d, an increase in blue fluorescence was observed, because apoptotic cells had increased membrane permeability, and thus Hoechst 33342 entered apoptotic cells more than normal cells, and fluorescence intensity was higher than that in normal cells. In addition, the structure of chromosomal DNA of apoptotic cells is changed so that the dye can bind to DNA more efficiently, and the p-glycoprotein pump function on apoptotic cell membranes is impaired, Hoechst 33342 cannot be efficiently discharged outside cells to be accumulated inside cells, and blue fluorescence is enhanced.
(4) The anticancer lipopeptide has high serum stability and low hemolytic toxicity
The hemolytic activity of the anti-cancer lipopeptides was evaluated by the effect of the anti-cancer lipopeptides on the morphology of mouse erythrocytes. As shown in FIGS. 4a-b, the morphology of the mouse erythrocytes in the normal saline group and the anticancer lipopeptide group is not changed much, and the mouse erythrocytes are in a normal biconcave disc shape, so the anticancer lipopeptide has low hemolytic toxicity and high biological safety.
A large proportion of anticancer peptides do not exert good antitumor effect in vivo because they have short biological half-life in vivo and low biological stability, and can be easily inactivated by hydrolysis by protease in vivo. Fetal Bovine Serum (FBS) is a blood component collected from cattle, and contains a plurality of proteolytic enzymes, and polypeptides are hydrolyzed by proteases to weaken the tumor-inhibiting activity. Therefore, lipopeptides were first pretreated with PBS and 10% Fetal Bovine Serum (FBS) -containing PBS, respectively, and whether the anticancer activity of lipopeptides was affected was examined by the MTT method. FIG. 4c is a comparison of the killing ability of P17 lipopeptide alone and after 24H of co-culture with serum to H22 cells, and it can be seen that the killing ability of lipopeptide after co-culture with serum to cancer cells is slightly reduced, but still has strong killing ability, indicating that the lipopeptide has good serum stability.
(5) The anticancer lipopeptide has good in vivo antitumor effect
To investigate the in vivo antitumor activity of the lipopeptides, a tumor-bearing mouse model was established by subcutaneous injection of approximately 1X 10 in the left axilla of mice 6 H22 cells, the tumor volume is 150-220mm 3 At that time, injection of lipopeptide drug was initiated, and tumor volume was measured and weighed. After the experiment is finished, the mouse is killed by a cervical dislocation method, the tumor tissue of the mouse is dissected, and then the mouse is fixed by 4% paraformaldehyde and is subjected to tissue section and HE staining treatment. As shown in fig. 5a, the relative tumor volume of mice in the anti-cancer lipopeptide group was significantly smaller than that in the normal saline group. As shown in FIG. 5b, the body weight of mice bearing tumor of the anticancer lipopeptide group did not change much with the increase of the administration times, which indicates that the anticancer lipopeptide has less toxic and side effects and can play the role of anti-tumor without affecting the body weight of the mice. To gain a more direct understanding of the anti-cancer effects of lipopeptides, tumor tissues were HE stained and observed under a microscope. As shown in FIGS. 5c-d, the tumor cell density of the anticancer lipopeptide group (FIG. 5d) was significantly decreased compared to the NS group (FIG. 5c) tumor tissues, and it can be seen that the anticancer lipopeptide has a good in vivo antitumor effect.
The above description is only a preferred example of the present invention and is not intended to limit the above facts. The invention is intended to cover such departures from the present disclosure as come within known or customary practice in the art to which this invention pertains.
Sequence listing
<110> Binzhou medical college
<120> preparation and anti-tumor effect of RGD/KLA integration lipopeptide
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 17
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 1
Arg Gly Asp Lys Leu Ala Lys Leu Ala Lys Lys Leu Ala Lys Leu Ala
1 5 10 15
Lys
Claims (3)
1. An anticancer lipopeptide, wherein the sequence from N-terminus to C-terminus is C 8 H 15 O-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Asp-Gly-Arg, wherein Lys is lysine, Ala is alanine, Leu is leucine, Asp is aspartic acid, Gly is glycine, Arg is arginine, C is 8 H 15 O represents caprylic acid, and the structural formula of the anticancer lipopeptide is as follows:
2. anticancer lipopeptide C 8 H 15 The preparation method of O-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Asp-Gly-Arg is characterized by comprising the following steps:
(1) synthesis of Fmoc-Lys (Boc) -Ala-Leu-Lys (Boc) -Asp (OtBu) -Gly-Arg (Pbf) -Wang Resin;
Fmoc-Lys (Boc) -Ala-Leu-Lys (Boc) -Ala-Leu-Lys (Boc) -Asp (OtBu) -Gly-Arg (Pbf) -Wang Resin samples were synthesized by solid phase synthesis; swelling Fmoc-Arg (Pbf) -Wang Resin with DMF for 30 min; detecting ninhydrin with a small amount of resin, and adding 20% piperidine-containing DMF solution to react for 30min to remove Fmoc protecting group if no color change exists; washing the resin with DMF, DCM and DMF sequentially, and swelling the resin with anhydrous DMF for 30 min; after swelling, adding 2 times of equivalent of amino acid and 2.6 times of equivalent of DCC, HOBT and DIEA for reaction for more than 48 h; after dialysis, a small amount of resin is taken for ninhydrin detection, and if no color change exists, the corresponding amino acid is connected; then, sequentially washing the resin with DMF, DCM, and DMF, dialyzing in 95% ethanol with dialysis bag (MW8000-14000) for more than 20 times, each time for 30 min; the above steps were repeated until Fmoc-Lys (Boc) -Ala-Leu-Lys (Boc) -Asp (OtBu) -Gly-Arg (Pbf) -Wang Resin samples were synthesized.
(2)C 8 H 15 Synthesis of O-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Asp-Gly-Arg;
first, Fmoc-Lys (Boc) -Ala-Leu-Lys (Boc) -Asp (OtBu) -Gly-Arg (Pbf) -Wang Resin is subjected to Fmoc group removal in the step (1) to obtain Lys (Boc) -Ala-Leu-Lys (Boc) -Ala-Leu-Lys (Boc) -Asp (OtBu) -Gly-Arg (Pbf) -Wang Resin, and freeze-drying the same; 2.5 times of equivalent of NHS and EDC and 2 times of equivalent of n-octanoic acid are put into anhydrous DMF for activation for 5h, and then the lyophilized resin is added for reaction for 48 h; after the reaction is finished, sequentially washing the resin with DMF, DCM and DMF, filling the resin into a dialysis bag (MW8000-14000), and dialyzing for more than 20 times, wherein the dialysis time is 30min each time; after the dialysis is finished, washing the dialysis membrane; the resulting sample was lyophilized and cleavage solution (TFA: H) was added 2 O: tis 95:2.5:2.5) cutting off side chain protecting group and resin to obtain target lipopeptide C 8 H 15 O-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Asp-Gly-Arg。
3. The anticancer lipopeptide C of claim 1 8 H 15 The application of O-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Lys-Ala-Leu-Lys-Ala-Leu-Lys-Asp-Gly-Arg in preparing antitumor drugs.
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