CN101870726A - Peptide - cisplatin conjugate and preparation method and application thereof - Google Patents
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Abstract
The invention discloses a peptide - cisplatin conjugate which is prepared by taking a peptide containing 12 amino acids as a carrier and coupling the pipetide with anticancer drug cisplatin, and the invention makes initial research on the pharmacological properties of the peptide - cisplatin conjugate. Cisplatin is widely applied to traditional cancer treatment, but the advantages of great toxic and side effect and the like thereof restrict the application thereof in the treatment. Therefore, to reduce the toxic and side effect of cisplatin is the key to improve the cisplatin medicine. The peptide - cisplatin conjugate obtained by the coupling of peptide and the cisplatin through chemical reaction has small molecular weight when being used for preparing anti-tumor disease medicine, has no antigenicity, does not cause allergic reaction, helps the medicine to display tumor targeting property, takes envoplakin, CD63 and other antigen as the targets, kills tumors with high efficiency, promotes the apoptosis of tumor cells and reduces the toxic and side effect.
Description
Technical field
The present invention relates to the preparation field of cancer therapy drug, be specifically related to one peptide species-cis-platinum conjugates and preparation method thereof and application.
Background technology
Cis-platinum, English name Cisplatin is nineteen sixty-five find first by people such as U.S. scientist B.Rosenborg first have the metal complexes of antitumour activity.He can cause this fact of mycelial growth of bacterium in unexpected discovery of nineteen sixty-five " inert platinum electrode ", thereby has launched the research to the anticancer property of cis dichloro two platinum.Cis-platinum is the center with divalence platinum with two chlorine atoms and two amino molecule bonded heavy metal complex, is similar to difunctional alkylating agent.Discover purine and the pyrimidine bases of the main site of action of cis-platinum at DNA, dna replication dna process that can anticancer, and damage structure on its cytolemma, the cis-platinum of high density can suppress RNA and proteinic synthetic, have stronger broad spectrum anticancer effect, especially evident in efficacy to genital system tumor to ovarian cancer and carcinoma of testis; To the incidence cancer, curative effect is preferably also arranged as nasopharyngeal carcinoma, thyroid carcinoma or laryngocarcinoma; Bladder cancer, lung cancer, malignant lymphoma, mammary cancer, renal cell carcinoma, prostate cancer, soft tissue sarcoma or malignant melanoma also there is certain curative effect; The pernicious ascites pleural fluid that cancer is caused has special efficacy.
But cis-platinum also can make the patient produce very strong untoward reaction, as bone marrow depression, gastrointestinal reaction, Toxicity of Kidney, neurotoxicity, anaphylaxis, electrolyte disturbance and hepatic disorder.And, because cis-platinum commonly used in the market and improvement complex compound thereof when acting on human body, do not have target, cause human body to produce the intensive untoward reaction.Therefore, the above-mentioned defective of cis-platinum especially its toxic side effect people are pressed for seek low toxicity more, efficient, good water solubility and do not have the platinum metals antitumor drug of the tumor-targeting of new generation of cross resistance.Cisplatin medicine is big because of its molecular weight at present, causes the immune response of human body easily, produces allergic symptom, so the cis-platinum complex compound of preparation antitumor drug also should possess less relatively this feature of molecular weight.
Summary of the invention
The object of the invention is according to the deficiencies in the prior art, provides that a kind of molecular weight is little, no antigen and have the polypeptide-cis-platinum conjugates of target.
Another purpose of the present invention is to provide the preparation method of aforementioned polypeptides-cis-platinum conjugates.
A further object of the invention is to provide the application of aforementioned polypeptides-cis-platinum conjugates in the medicine of preparation treatment cancer and complication thereof.
Above-mentioned purpose of the present invention is achieved by the following technical programs:
Polypeptide of the present invention-cis-platinum conjugates is to be formed by polypeptide with general formula shown in the formula (I) and cis-platinum coupling:
Leu-Leu-Ala-Asp-Thr-Thr-His-His-Arg-X-Y-Z
(I)；
Wherein, X is Ala, Pro or Arg;
Y is Trp, Tyr or Phe;
Z is Cys, Thr or Ser.
In aforementioned polypeptides-cis-platinum conjugates, X is preferably Arg or Pro; Y is preferably Phe or Tyr; Z is preferably Cys or Ser.
In aforementioned polypeptides-cis-platinum conjugates, X most preferably is Arg, and Y most preferably is Phe, and Z most preferably is Cys.
In aforementioned polypeptides-cis-platinum conjugates, described polypeptide be acetylizad polypeptide or with the polypeptide of polyoxyethylene glycol copolymerization.
The preparation method of polypeptide of the present invention-cis-platinum conjugates comprises the steps:
(1) by the synthetic polypeptide of solid-phase polypeptide synthesis method;
(2) will synthesize good polypeptide and alkylene dihalide and react, obtain corresponding peptide C-end halogenide;
(3) peptide C-end halogenide and two (1-Ft amino) ethylamine carries out condensation reaction, obtains amido modified polypeptide precursor, shown in (II):
(4) the polypeptide precursor that obtains in the step (3) reduces deprotection reaction, obtains the polyamines modified peptides, shown in (III):
(5) with the polyamines modified peptides and the platinum chloride reaction of gained in the step (4), obtain polypeptide-cis-platinum conjugates.
As a kind of preferred version, among the above-mentioned preparation method, the halogen element in the described alkylene dihalide is Br or I.
As a kind of preferred version, among the above-mentioned preparation method, the synthetic polypeptide of described solid-phase polypeptide synthesis method comprises the steps:
(1) introducing of first Pro: Fmoc-Pro-OH is covalently bound on the CTC resin, Fmoc-Pro-OH/Resin=1.1, and reaction 4h obtains Fmoc-Pro-CTC Resin;
(2) all the other amino acid whose importings: all the other amino acid are covalently bound on the CTC resin, and Fmoc-AA/Resin=3.0 adds dimethyl formamide solution, and coupling agent is the polypeptide condensing agent;
(3) remove Fmoc: remove by the dimethyl formamide solution that adds 20 volume % piperidines;
(4) resin cracking: the trifluoroacetic acid solution that adds 1 volume % is removed the CTC resin, obtains polypeptide.
Wherein, the reaction capacity of described CTC resin is 0.9mmol/g.
Among the present invention, the reason of the synthetic polypeptide of preferred solid-phase polypeptide synthesis method is: in the method for the synthetic polypeptide of solution method mass-producing commonly used, though the easy purifying of product, purity also can be controlled, and its shortcoming is that reactions steps is too many, and preparation cycle is long; Adopt the solid-phase polypeptide synthesis method, the sample of some amount can be provided within a short period of time.Specifically, at polypeptide of the present invention, use full-automatic solid-phase polypeptide synthesizer, can prepare about 10～15g sample in all left and right sides time, purity is generally more than 85%.Be separated (about 1～2 week) through preparative liquid, purity can reach 98% again, and yield is about 40～60%.
In addition, the function of polypeptide effect and mechanism can be ever-changing, from as the immunostimulation hormone, assist protein degradation, to having anti-microbial activity.In general, polypeptide be proved to be have higher activity and highly selective, lower toxicity, lower accumulation in organ, and the complication of less drug interaction.Shortcoming is that its bioavailability is poor, degrade easily and synthesize the difficulty.The factor that influences polypeptide drugs stability mainly is that in the time of in medicine enters body, it is small-molecular peptides or amino acid that various lytic enzymes cause drug degradation.In recent years, the medicine-feeding technology of polypeptide drug also had develop rapidly.Tradition protein/polypeptide class medicine mostly is injection type greatly, use rather inconvenience, new technology comprises oral preparation/long-acting sustained-release agent, microcapsule, Foradil Aerolizer formoterol fumarate at present, PEG chemical preparation (polyoxyethylene glycol embedding medium) or the like, thus protein/polypeptide class medicine transformation period and bioavailability in vivo improved greatly.In addition, we will utilize the chemically modified means to improve polypeptide stability in vivo, concrete modifying method can but be not limited to following method:
1. the acetylize of nitrogen end: free amino group and diacetyl oxide or acyl chloride reaction with the Polypetide Nitrogen end, make it change into the acetyl peptide, can improve the polypeptide drugs transformation period in vivo usually.
2. the easily amino acid whose replacement of enzymolysis: whole polypeptide drugs of being made up of L-amino acid, common stability is all poor.If with some L-amino acid replacement to the enzyme sensitivity is D-amino acid, particularly change the amino acid whose configuration in polypeptide two ends, its stability may obviously be improved.Except using D-amino acid, other are such as hydroxy-amino-acid, and the amino acid that methylates, heterocyclic amino acid etc. also can be used for the amino acid whose replacement of L-.
3. and polyethylene glycol polymeric: in general, be easy to be discharged by the metabolism of kidney for small-molecule drug (M＜5000).Therefore,, both can improve the metabolic stability of kidney, can increase the steric hindrance of enzymic hydrolysis again by being connected with PEG (MW＞10000).And PEG itself has and has no side effect, advantage such as water-soluble and good biocompatibility.
Polypeptide of the present invention-cis-platinum conjugates can be used for preparing the medicine for the treatment of cancer, in particular for the medicine of preparation treatment lung cancer (comprising small cell lung cancer SCLC and nonsmall-cell lung cancer NSCLC), lung cancer ascites pleural fluid, colorectal carcinoma, the esophageal carcinoma or genital system tumor.
Polypeptide of the present invention-cis-platinum conjugates is the extension of the original effect of cis-platinum, its antitumor characteristic is the function of cis-platinum, but the effect of used polypeptide is the performance tumor-targeting, can be target spot with antigens such as envoplakin, CD63 respectively, tumour cell there is efficient lethality, can promotes the apoptosis of tumour cell.
Compared with prior art, the present invention has following beneficial effect:
(1) molecular weight of polypeptide of the present invention-cis-platinum conjugates is less than 2000, and far below the molecular weight (being generally 100,000～150,000) of antibody molecule, therefore no antigen almost can not cause body to produce immune anaphylaxis.
(2) polypeptide of the present invention-cis-platinum conjugates is better than macromolecular substance such as antibody to the tissue penetration and the tissue distribution effect of solid tumor;
(3) polypeptide of the present invention-cis-platinum conjugates has tumor-targeting, avoids the position beyond tumour to gather, and reduces untoward reaction, has increased the specificity of tumor-killing;
(4) toxic side effect of polypeptide of the present invention-cis-platinum conjugates is low, is 1/20～1/50 of original cis-platinum, and the curative effect of same dose medication has also strengthened 10～20 times.
Embodiment
Further explain the present invention below in conjunction with embodiment, but embodiment does not do any type of qualification to the present invention.
The preparation of embodiment 1 polypeptide-cis-platinum conjugates
Add 10 milliliters of DMF in 25 milliliters of round-bottomed flasks, 25mg (0.017mol) LLADTTHHRPWT stirred 24 hours under 3.5mg (0.019mmol) glycol dibromide and the anhydrous sodium carbonate 15mg.. room temperature.Add two (2-phthalic imidine) ethamine 10mg (0.028mol), continue to stir 24 hours.1 milliliter of the hydrazine aqueous solution of adding 80% continues to stir 3 days.Add 10 milliliter of 70% acetone soln, filter.With 70% acetone soln washing precipitation.Resolution of precipitate in 3 ml waters, is added 4.5 milligrams of platinous chlorides (0.017mmol), the solution lyophilize is obtained product.
Anti-lung cancer pharmacodynamic study in the body of embodiment 2 polypeptide-cis-platinum conjugates
1 materials and methods
1.1 cell
The NCI-H1299 human lung carcinoma cell line is provided by Chinese Academy of Sciences's Shanghai cell research.
1.2 laboratory animal
The healthy male Balb-c nude mice of SPF level, in 5～6 ages in week, body weight 16～20g is provided by Guangdong Medical Lab Animal Center, conformity certification number: SCXK (Guangdong) 2008-0002.In Guangdong Pharmaceutical University's medicine academy of sciences new medicament screen and pharmacodynamics assessment centers IVC system, 25 ℃ of temperature, humidity 70～80% is raised and is used for experiment after 1 week.
1.3 main agents and medicine
High sugared DMEM substratum is available from Gibco company; The import foetal calf serum is available from Hyclone company; Be subjected to reagent product: H001, the injection liquid self-control; The positive control medicine: cis-platinum, produce lot number: BXF 23E3 by Bayer A.G.
1.4 cell cultures with go down to posterity
Human lung carcinoma cell NCI-H1299 cultivates with the high sugared DMEM nutrient solution that contains 10% foetal calf serum, places 37 ℃, 5%CO
2Cultivate in the constant temperature incubator, cell is the monolayer adherence growth, treats that the attached cell fusion reaches at 80%～90% o'clock, goes down to posterity with 0.25% tryptic digestion that contains 0.01%EDTA.
1.5 the foundation of lotus people lung cancer nude mice animal model
To be in logarithmic phase, human lung carcinoma cell NCI-H1299 in good condition, after 0.25% trysinization that contains 0.01%EDTA, the centrifugal 1min of 1000rpm, remove supernatant, add the resuspended precipitation of the high sugared DMEM nutrient solution of serum-free, cell density is adjusted in cell counting, chooses an injection site in nude mice back of the body belly, subcutaneous vaccination human lung carcinoma cell NCI-H1299, each site injection cell about 2 * 10
6Individual, after naked eyes will see that the nude inoculation position has tumour to grow 7 day after tomorrow, can be used for experiment.
1.6 GP TH
Lotus people lung cancer nude mice is divided into 4 groups at random, is provided with as follows respectively: (1) negative control group (physiological saline); (2) positive drug (cis-platinum) control group is (10mg/kg); Three groups of investigational agents (polypeptide-cis-platinum conjugates) are respectively: (3) polypeptide-cis-platinum conjugates low dose group (2.5mg/kg), dosage group (5mg/kg) in (4) polypeptide-cis-platinum conjugates, (5) polypeptide-cis-platinum conjugates high dose group (10mg/kg).Each organizes nude mice intraperitoneal injection every day once, and the administration volume is 0.1ml/10g, successive administration 14d.
1.7 observe tumor growth situation and nude mice generalized case
Respectively at the 1st, 4,7,10,14 days weighing nude mice body weight of administration, with the 1st, 7,14 days tumour sizes of vernier caliper measurement administration, calculate tumor size with following formula: V=ab2 π/60, a: the maximum diameter of tumor nodule, b: the transverse diameter of tumor nodule, take off neck after last is measured and put to death nude mice, separate tumor tissues and take pictures, weigh, survey volume, respectively with 4% Paraformaldehyde 96,10% formalin fixed tumor tissues.
1.8 polypeptide-cis-platinum conjugates is at the tissue distribution figure of tumor bearing nude mice
Get fluorescein FITC labeling polypeptide (Fitc-polypeptide-cis-platinum conjugates) 1mg and be dissolved in 600uLPBS, be made into 1.67mg/mL.Dilute 4 times and carry out tail vein injection.(10:00) beginning cardiac perfusion behind the 15min.(fixedly mouse is opened the thoracic cavity, exposes heart, and the perfusion device enters left ventricle from apex, opens an osculum then in the right atrium, washes blood vessel with 30mLNS earlier, then with the 50mL Paraformaldehyde 96 first quick and back slow perfusion carry out fixation of tissue).Finish perfusion, get started to dissect and get the heart of mouse, liver, spleen, lung, kidney and tumor tissues.Carry out frozen section (order is the heart, liver, spleen, lung, kidney, tumor tissues), carry out frozen section, take pictures (all photos are 100x and 200x, and exposure time is 10s).
1.9 data processing
Data results with
Expression adopts SPSS 11.0 statistical softwares to carry out the check of two independent sample t, and there is statistical significance P＜0.05 for difference.
2 results
2.1 tumor formation rate
This experiment all about 0.15cm size tumor tubercle occurred at inoculation position in 3 days with nude mice after human lung carcinoma cell NCI-H1299 subcutaneous vaccination, tumor formation rate reaches 95%, shows successfully to set up lotus people lung cancer nude mice model.
2.2 respectively organize nude mice generalized case and body weight change
Phenomenon such as tangible movable minimizing, delay of response, spiritual atrophy all do not occur during most nude mices treatments, become thin.The body weight change situation of respectively organizing nude mice after the administration is as shown in table 1.
Table 1 is respectively organized nude mice body weight (g)
2.2 tumor growth situation
2.2.1 volume change: with the 1st, 7,14 days nude mice tumour lines of apsides of vernier caliper measurement administration size, obtain each gross tumor volume value, the result is as shown in table 2; According to formula " Δ V
1Gross tumor volume value after the=administration-administration pre-neoplastic volume cost " obtain the changing value that the nude mice gross tumor volume was respectively organized in administration in the 7th, 14 days respectively, the result is as shown in table 3; According to formula " Δ V
2=medicine group gross tumor volume value-negative control group gross tumor volume value " obtain the gross tumor volume difference that the 7th, 14 days each medicine groups of administration are compared with negative control group, the result is as shown in table 4.As seen, the nude mice gross tumor volume of positive drug group is significantly less than negative control group and polypeptide-cis-platinum conjugates group after the administration, and difference has statistical significance (P＜0.01) between the negative control group, the nude mice gross tumor volume of polypeptide-cis-platinum conjugates high dose group is less than negative control group, and difference has statistical significance (P＜0.01) between the negative control group, and in polypeptide-cis-platinum conjugates, the nude mice gross tumor volume of low dose group is slightly less than negative control group, but compares not statistically significant (P＞0.05) with control group.
Table 2 is respectively organized nude mice gross tumor volume (mm
3)
*P＜0.05,
*P＜0.01, medicine group vs negative control group.
Compare before table 3 and the administration, the changing value (mm of nude mice gross tumor volume was respectively organized in administration in the 7th, 14 days
3)
*P＜0.05,
*P＜0.01, medicine group vs negative control group.
After table 4 administration, the gross tumor volume difference (mm that each medicine group is compared with negative control group
3)
2.2.2 knurl heavily changes: after treatment in the 14th day finishes, put to death tumor bearing nude mice, take out the knurl piece, weighing results sees Table 5.The nude mice tumor weight of positive drug group obviously is lighter than negative control group and polypeptide-cis-platinum conjugates group after the administration, and difference has statistical significance (P＜0.01) between the negative control group, the nude mice tumor weight of polypeptide-cis-platinum coupling object height, middle dosage group is lighter than negative control group, and the difference between the negative control group has statistical significance (P＜0.05).Obtain the tumor weight difference that the 7th, 14 days each medicine groups of administration are compared with negative control group according to formula " the average knurl of Δ W=medicine group heavily be worth-the average knurl of negative control group heavily be worth ", the result is as shown in table 6.
Table 5 is respectively organized nude mice tumor weight (g)
*P＜0.05,
*P＜0.01, medicine group vs negative control group.
After table 6 administration, the knurl method of double differences value (g) that each medicine group is compared with negative control group
2.2.3 after treatment in the 14th day finishes, put to death tumor bearing nude mice, take out the knurl piece, the gross tumor volume after the measurement of water yield method is peeled off the results are shown in Table 7.The nude mice gross tumor volume of positive drug group is significantly less than negative control group and H001 group after the administration, and difference has statistical significance (P＜0.01) between the negative control group, the nude mice gross tumor volume of H001 high dose group is less than negative control group, and difference has statistical significance (P＜0.01) between the negative control group, and in polypeptide-cis-platinum conjugates, the nude mice gross tumor volume of low dose group and negative control group approaching, not statistically significant.Obtain the gross tumor volume difference that the 7th, 14 days each medicine groups of administration are compared with negative control group according to formula " Δ V=medicine group gross tumor volume value-negative control group gross tumor volume value ", the result is as shown in table 8.
Gross tumor volume (cm after the measurement of table 7 water yield method is peeled off
3)
P＜0.05,
*P＜0.01, medicine group vs negative control group.
Gross tumor volume difference (the cm that gross tumor volume after the measurement of table 8 water yield method is peeled off, each medicine group are compared with negative control group
3)
2.2.4 fluorescein-labeled polypeptide-cis-platinum conjugates is at the intravital tissue distribution figure of tumor bearing nude mice, polypeptide-cis-platinum conjugates has more distribution in the cancerous lung tissue of visible inoculation, and the heart, liver, brain, lung, kidney healthy tissues the results are shown in Figure 1.
3 conclusions
Utilize people's lung cancer NCI-H1299 cell, be inoculated in nude mice, successfully duplicated model of nude mice bearing tumor.To being subjected to reagent polypeptide-cis-platinum conjugates to carry out the test of antitumor drug effect, the result shows: when polypeptide-cis-platinum conjugates can significantly suppress tumor growth during for 10mg/kg, compare P＜0.05 with contrast model group (negative control group); When polypeptide-when the cis-platinum conjugates is 2.5mg/kg and 5mg/kg, not obvious to the restraining effect of NCI-H1299 lung cancer growth, compare not statistically significant with control group.The result shows, when polypeptide-cis-platinum conjugates dosage has the effect of obvious anti-people's lung cancer during more than or equal to 10mg/kg.
The pharmacodynamic experiment research of embodiment 3 polypeptide-cis-platinum conjugates Tuberculosis in vitro intestinal cancer
1 materials and methods
1.1 cell strain
The strain of SW1116 human colon cancer cell is provided by Chinese Academy of Sciences's Shanghai cell research.
1.3 main agents and medicine
High sugared DMEM substratum is available from Gibco company; The import foetal calf serum is available from Hyclone company; Be subjected to reagent product: H001, the injection liquid self-control; The positive control medicine: cis-platinum, produce lot number: BXF 23E3 by Bayer A.G.
1.4 cell cultures with go down to posterity
Human colon cancer cell SW1116 cultivates with the high sugared DMEM nutrient solution that contains 10% foetal calf serum, places 37 ℃, 5%CO
2Cultivate in the constant temperature incubator, cell is the monolayer adherence growth, treats that the attached cell fusion reaches at 80%～90% o'clock, goes down to posterity with 0.25% tryptic digestion that contains 0.01%EDTA.
Adopt the MTT colorimetry, choose logarithmic phase SW1116 cell, make cell suspension, adjusting cell density is 1 * 10
5/ L adds the MnSOD that final concentration is 0.5～10mg/L respectively in 96 porocyte culture plates
mThe CO2 incubator is put in 100 μ l/ holes, at 37 ℃, 5%CO
2And cultivate 24～72h under the saturated humidity condition, and add MTT solution (5g/L) 10 μ l/ holes from cultivating terminal point 4h, continue to cultivate 4h, add 10%SDS (0.01mol/L HCl solution) 100 μ l/ holes, standing over night.Microplate reader (Micro-TEKWAVE X) is surveyed absorbancy (A) value in the 570nm place, calculate cell proliferation inhibition rate and half-inhibition concentration (IC50).
1.5 GP TH
Human colon cancer cell is divided into 6 groups at random, is provided with as follows respectively: (1) negative control group (physiological saline); (2) positive drug (cis-platinum) control group is (10ug/ml); Five groups of investigational agents (polypeptide-cis-platinum conjugates) are respectively: (3) polypeptide-cis-platinum conjugates 5ug/ml dosage group, (4) polypeptide-cis-platinum conjugates 10ug/ml dosage group, (5) polypeptide-cis-platinum conjugates 20ug/ml dosage group, (6) polypeptide-cis-platinum conjugates 20ug/ml dosage group.Each organizes nude mice intraperitoneal injection every day once, and the administration volume is 0.1ml/10g, successive administration 14d.
2 results
The IC50 of H001 resistive connection intestinal cancer SW1116 is 18ug/ml, and the IC50 of cis-platinum resistive connection intestinal cancer SW1116 is 15ug/ml.
3 conclusions
Polypeptide-cis-platinum conjugates has the lethal effect the same with cis-platinum to human colon cancer cell, and its IC50 is respectively 18ug/ml and 15ug/ml.
Claims (10)
1. one peptide species-cis-platinum conjugates is characterized in that described polypeptide-cis-platinum conjugates is formed by polypeptide with general formula shown in the formula (I) and cis-platinum coupling:
Leu-Leu-Ala-Asp-Thr-Thr-His-His-Arg-X-Y-Z
(I)；
Wherein, X is Ala, Pro or Arg;
Y is Trp, Tyr or Phe;
Z is Cys, Thr or Ser.
2. polypeptide according to claim 1-cis-platinum conjugates is characterized in that described X is Arg.
3. polypeptide according to claim 1-cis-platinum conjugates is characterized in that described Y is Phe.
4. polypeptide according to claim 1-cis-platinum conjugates is characterized in that described Z is Cys.
5. according to the described polypeptide of any claim-cis-platinum conjugates in the claim 1～4, it is characterized in that described polypeptide be acetylizad polypeptide or with the polypeptide of polyoxyethylene glycol copolymerization.
6. the preparation method of the described polypeptide of any claim-cis-platinum conjugates in the claim 1～4 is characterized in that comprising the steps:
(1) by the synthetic polypeptide of solid-phase polypeptide synthesis method;
(2) will synthesize good polypeptide and alkylene dihalide and react, obtain corresponding peptide C-end halogenide;
(3) peptide C-end halogenide and two (1-Ft amino) ethylamine carries out condensation reaction, obtains amido modified polypeptide precursor, shown in (II):
(4) the polypeptide precursor that obtains in the step (3) reduces deprotection reaction, obtains the polyamines modified peptides, shown in (III):
(5) with the polyamines modified peptides and the platinum chloride reaction of gained in the step (4), obtain polypeptide-cis-platinum conjugates.
7. the preparation method of polypeptide according to claim 6-cis-platinum conjugates is characterized in that in the step (1), and the synthetic polypeptide of described solid-phase polypeptide synthesis method comprises the steps:
(1) introducing of first Pro: Fmoc-Pro-OH is covalently bound on the CTC resin, Fmoc-Pro-OH/Resin=1.1, and reaction 4h obtains Fmoc-Pro-CTC Resin;
(2) all the other amino acid whose importings: all the other amino acid are covalently bound on the CTC resin, and Fmoc-AA/Resin=3.0 adds dimethyl formamide solution, and coupling agent is the polypeptide condensing agent;
(3) remove Fmoc: remove by the dimethyl formamide solution that adds 20 volume % piperidines;
(4) resin cracking: the trifluoroacetic acid solution that adds 1 volume % is removed the CTC resin, obtains polypeptide.
8. the preparation method of polypeptide according to claim 6-cis-platinum conjugates is characterized in that in the step (2), the halogen element in the described alkylene dihalide is Br or I.
9. the preparation method of polypeptide cis-platinum conjugates according to claim 8, the reaction capacity that it is characterized in that described CTC resin is 0.9mmol/g.
10. the application of the described polypeptide of claim 1～4-cis-platinum conjugates is characterized in that described polypeptide-cis-platinum conjugates is used to prepare the medicine of treatment cancer or cancer complication.
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| WO2024041535A1 (en) * | 2022-08-22 | 2024-02-29 | 赣州和美药业股份有限公司 | Nano-composition, preparation method therefor, and use thereof |
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| CN114831134A (en) * | 2022-01-05 | 2022-08-02 | 北京化工大学 | Platinum nano particle/short peptide hydrogel and preparation method and antibacterial application thereof |
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