CN107892710B - Supported platinum complex oxidant easy to recycle and use and preparation method and application thereof - Google Patents
Supported platinum complex oxidant easy to recycle and use and preparation method and application thereof Download PDFInfo
- Publication number
- CN107892710B CN107892710B CN201710934846.6A CN201710934846A CN107892710B CN 107892710 B CN107892710 B CN 107892710B CN 201710934846 A CN201710934846 A CN 201710934846A CN 107892710 B CN107892710 B CN 107892710B
- Authority
- CN
- China
- Prior art keywords
- platinum complex
- reaction
- oxidant
- solid
- supported platinum
- 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.)
- Expired - Fee Related
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Peptides Or Proteins (AREA)
- Catalysts (AREA)
Abstract
The invention discloses a supported platinum complex oxidant easy to recycle. The supported platinum complex oxidant has a structure shown as a formula (I). The invention also provides a preparation method of the oxidant, which comprises the following steps: a) reacting polystyrene microspheres with p-toluenesulfonyl chloride by using pyridine as a solvent to obtain yellow solid; b) ultrasonically dispersing the obtained yellow solid in ethylenediamine, and stirring to react to obtain a dark yellow solid; c) taking DMF as a solvent, and stirring the obtained dark yellow solid and a divalent platinum complex to react to obtain a black solid; d) and ultrasonically dispersing the black solid in ethanol, and then fully reacting with N-bromosuccinimide to obtain a light black solid, namely the supported platinum complex oxidant. The preparation method of the oxidant is simple, easy to recycle and high in reaction yield.
Description
Technical Field
The invention relates to a supported platinum complex oxidant, in particular to a supported platinum complex oxidant easy to recycle and a preparation method and application thereof.
Background
The polypeptide intramolecular disulfide bond is an important functional group for maintaining the high-order conformation of the polypeptide and improving the physiological activity and the pharmacological activity of the polypeptide. Disulfide bonds in drug polypeptide molecules can improve the stability and permeability of the drug molecules in vivo and increase the specific binding of the drug molecules to target molecules, thereby improving the biological activity of the polypeptide drug(s) ((Nature Chem., 2014, 6, 1009–1016; Chem. Rew., 2013, 114, 901-926.). A plurality of polypeptides comprising intramolecular disulfide bondsDrugs have been approved by the FDA and used in the treatment of various diseases, such as: octreotide for treating various endocrine tumors, nesiritide for treating heart failure, calcitonin for treating Paget's disease, oxytocin for uterine contraction, eptifibatide for resisting platelet drug, pramlintide for treating diabetes, and other polypeptide drugs. At present, a large number of polypeptides containing intramolecular disulfide bonds have been developed, and the development of polypeptide drugs has been promoted (Future Med. Chem., 2009, 1, 361-377.). The generation of disulfide bonds in polypeptide drugs is an important reaction in fine organic chemistry and pharmaceutical chemistry.
The traditional method for synthesizing polypeptide intramolecular disulfide bonds is as follows: (I) through solid-liquid reaction, polypeptide intramolecular disulfide bond is directly formed on the solid phase carrier. The solid-liquid reaction is carried out under the quasi-dilution condition, so that the formation of intramolecular disulfide bonds is promoted, and the formation of intermolecular disulfide bonds of the byproduct polypeptide is reduced. However, the yield of the polypeptide containing intramolecular disulfide bonds is greatly affected by the loading amount of the solid phase carrier, and the yield is not ideal. (II) oxidative folding to form intramolecular disulfide bonds in the liquid phase by homogeneous reaction: (II)Eur. J. Org. Chem., 2014, 3519–3530.）。
Currently, oxidants used for intramolecular disulfide bond synthesis of polypeptide drugs include: iodine, thallium (III) trifluoroacetate, oxygen, DMSO, oxidized glutathione, Ellman's reagent, and the like. However, when these oxidizing agents are used to synthesize disulfide bonds in polypeptide molecules, some oxidizing agents may react with methionine, tyrosine, and tryptophan residues in polypeptide chains, some oxidizing agents may produce by-products of dimers or multimers of polypeptides, which results in a decrease in yield, and the oxidizing agents may not be recycled, and some oxidizing agents may produce by-products with high toxicity(s) ((ii) (iii))Eur. J. Org. Chem., 2014, 3519–3530; RSC Adv. 2014, 4, 13854–13881; Synthesis, 2008, 16, 2491–2509.) 。
Besides the common oxidants, the tetravalent platinum complex oxidant is widely applied to the synthesis of intramolecular disulfide bonds of drugs, and the oxidation effect is superior to that of other currently used oxidants (A)J. Am. Chem. Soc. 2000, 122, 6809–6815; J. Org. Chem. 1999, 64, 4590–4595; Bioorg.Med. Chem. Lett. 2002, 12, 2237-2240.). However, the application of the platinum complex is limited due to high price, difficult preparation and the like, so if a platinum complex oxidant which is easy to recycle can be prepared, the cost expenditure for synthesizing the intramolecular disulfide bond of the drug by using the oxidant can be greatly reduced.
Disclosure of Invention
The invention aims to provide a supported platinum complex oxidant easy to recycle and use and application thereof, so as to solve the problems that the existing platinum complex oxidant cannot be recycled or the recycling operation is complex when used for synthesizing polypeptide intramolecular disulfide bonds.
The second purpose of the invention is to provide a preparation method of the supported platinum complex oxidant which is easy to recycle.
One of the objects of the invention is achieved by:
a supported platinum complex oxidant easy to recycle is a polystyrene microsphere loaded with a tetravalent platinum complex, and has a structure shown as a formula (I):
in the structure shown in formula (I), M is Pt4+Y is Br－And the particle size of the polystyrene microspheres is 75-320 mu m.
The polystyrene microsphere is a polyethylene glycol modified polystyrene microsphere.
The particle size of the polystyrene microspheres is 110 mu m.
The invention also provides application of the supported platinum complex oxidant in preparation of polypeptides containing intramolecular disulfide bonds.
The application can be realized by the following technical scheme:
carrying out oscillation reaction on a supported platinum complex oxidant and a reduced polypeptide solution for 0.5-1 h at room temperature, filtering and separating after the reaction is finished, and respectively recovering a solid obtained by filtering and a filtrate containing a reaction product; separating the reaction product in the filtrate by high performance liquid chromatography; and regenerating the supported platinum complex oxidant after the regeneration treatment of the solid obtained by filtering, and recycling.
The dosage ratio of the supported platinum complex oxidant to the reduced polypeptide is preferably that the mole ratio of the tetravalent platinum complex to the reduced polypeptide is 1-1.2: 1.
The reduced polypeptide solution is prepared by taking reduced polypeptide as a solute and water as a solvent.
The reduced polypeptide is a reduced polypeptide containing sulfydryl and capable of forming intramolecular disulfide bonds, and specifically comprises reduced polypeptide (1), reduced RGD peptide, reduced oxytocin, reduced arginine vasopressin and reduced oxytocin. The amino acid sequence of the reduced peptide (1) is as follows: H-Cys-Gly-Tyr-Cys-His-Lys-Leu-His-Gln-Met-NH2(ii) a The amino acid sequence of the reduced RGD peptide is as follows: H-Cys-Arg-Gly-Asp-Lys-Gly-Pro-Asp-Cys-NH2(ii) a The amino acid sequence of the reduced oxytocin is as follows: H-Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Lys-Gly-NH2(ii) a The amino acid sequence of the reduced arginine vasopressin is as follows: H-Cys-Tyr-Phe-Gln-Asn-Cys-Pro-Arg-Gly-NH2(ii) a The amino acid sequence of the reduced oxytocin is as follows: H-Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly-NH2。
The second purpose of the invention is realized by the following steps:
a preparation method of a supported platinum complex oxidant easy to recover and recycle comprises the following steps:
a) taking pyridine as a solvent, and reacting the polystyrene microspheres and p-toluenesulfonyl chloride for 2-3 d at room temperature according to the mass ratio of 1: 3-5;
after the reaction is finished, filtering, washing the obtained solid with ethanol, and then drying in vacuum to obtain a yellow solid;
b) ultrasonically dispersing the obtained yellow solid in ethylenediamine, and stirring and reacting for 1-2 d at the temperature of 60-80 ℃;
after the reaction is finished, cooling to room temperature, filtering, washing the obtained solid with ethanol, and drying in vacuum to obtain a dark yellow solid;
c) taking DMF as a solvent, and stirring and reacting the obtained dark yellow solid and a divalent platinum complex for 5-8 hours at the temperature of 70-90 ℃ according to the mass ratio of 1: 0.1-0.5;
after the reaction is finished, filtering and separating, washing the obtained solid, and then drying in vacuum to obtain a black solid;
d) ultrasonically dispersing the black solid in ethanol, and then fully reacting with N-bromosuccinimide at room temperature;
and after the reaction is finished, filtering and separating, washing the obtained solid, and drying in vacuum to obtain a light black solid, namely the supported platinum complex oxidant.
In the preparation method of the supported platinum complex oxidant, in the step (a), the polystyrene microsphere is a polyethylene glycol modified polystyrene microsphere. The particle size of the polystyrene microspheres is 75-320 mu m.
In the step (c), the divalent platinum complex is cis-Pt (en) Cl2。
In the preparation method of the supported platinum complex oxidant, in the step (d), the dosage of the N-bromosuccinimide is not less than 2 times of the dosage of the divalent platinum complex by mol.
The oxidizing agent and its preparation of the present invention have many advantages:
1. the supported platinum complex oxidant is simple to prepare and can be recycled after regeneration treatment, and the yield of the synthesized oxidized polypeptide is not influenced by the recycling frequency of the oxidant.
2. Compared with the existing oxidant, the supported platinum complex oxidant has the advantage of high yield.
3. After the polypeptide intramolecular disulfide bond is synthesized, the recovery and regeneration treatment of the oxidant are simple, and the supported platinum complex oxidant takes water as a reaction solvent, and the aim of separating a product can be achieved by filtering after the reaction.
Drawings
FIG. 1 is a flow chart of the reaction for preparing the supported platinum complex oxidant.
FIG. 2 is an SEM scan of a supported platinum complex oxidant of the present invention.
FIG. 3 is a comparison of chromatograms of reduced polypeptide (1) before and after reaction with an oxidant supported platinum complex of the present invention.
FIG. 4 is a mass spectrum of oxidized polypeptide (1).
FIG. 5 is a comparison of chromatograms of reduced RGD peptide before and after reaction with the supported platinum complex oxidant of the present invention.
FIG. 6 is a mass spectrum of oxidized RGD peptide.
FIG. 7 is a comparison of chromatograms of reduced oxytocin before and after reaction with a supported platinum complex oxidant of the present invention.
Figure 8 is a mass spectrum of oxidized oxytocin.
FIG. 9 is a comparison of chromatograms of reduced arginine vasopressin before and after reaction with the supported platinum complex oxidant of the present invention.
FIG. 10 is a mass spectrum of oxidized arginine vasopressin.
FIG. 11 is a comparison of chromatograms of reduced oxytocin before and after reaction with a supported platinum complex oxidant of the present invention.
Figure 12 is a mass spectrum of oxidized oxytocin.
Detailed Description
FIG. 1 shows a reaction scheme for preparing a supported platinum complex oxidant according to the present invention. As shown in figure 1, the invention makes polyethylene glycol modified polystyrene microsphere react with paratoluensulfonyl chloride, and then react with ethylenediamine (nitrogen is introduced) to obtain ethylenediamine modified polystyrene microsphere (which is dark yellow solid), and the ethylenediamine modified polystyrene microsphere and cis-form ethylenediamine dichloride platinum (cis-form Pt (en) Cl)2) Reacting to obtain polystyrene microspheres (black solids) loaded with divalent platinum complexes, and carrying out oxidation reaction on the polystyrene microspheres loaded with divalent platinum complexes and N-bromosuccinimide to obtain polystyrene microspheres (light black solids) loaded with tetravalent platinum complexes, namely the supported platinum complex oxidant, wherein the specific preparation process is as shown in examples 1-3:
example 1
The polystyrene microspheres used in this example were commercially available and had a particle size of 110 μm.
Adding 1.0g of polystyrene microspheres and 100mL of anhydrous pyridine into a 250 mL three-neck flask, then adding 5g of paratoluensulfonyl chloride, magnetically stirring, and reacting at room temperature for 3 d; after the reaction was completed, filtration was carried out, and the obtained solid was washed with ethanol 5 times and then 50 timesoC, vacuum drying for 12 hours to obtain yellow p-toluenesulfonyl modified polystyrene microspheres;
② adding 1.0g of p-toluenesulfonyl modified polystyrene microspheres into a 100mL three-necked flask, then adding 50mL of ethylenediamine, magnetically stirring, 60 percentoC, reacting for 2 d; after the reaction is finished, filtering, washing the solid with ethanol for 3 times, and then drying in vacuum to obtain dark yellow ethylenediamine modified polystyrene microspheres;
③ 0.2g of the ethylenediamine modified polystyrene microspheres are weighed into a 100mL three-necked flask, 30mL of DMF is added, and then 0.1g of cis-Pt (en) Cl is added2Ultrasonic dispersion, magnetic stirring, 90oC, reacting for 5 hours; after the reaction is finished, filtering and separating, washing the solid with DMF for 3 times, then washing with ethanol for 3 times, and then drying in vacuum overnight to obtain black polystyrene microspheres loaded with divalent platinum complexes;
and fourthly, weighing 0.1g of polystyrene microsphere loaded with the divalent platinum complex into a 100mL three-necked bottle, adding 50mL of ethanol, then adding 5g N-bromosuccinimide, magnetically stirring, reacting at room temperature for 12h, filtering and separating after the reaction is finished, washing the solid with ethanol for 3 times, and then drying in vacuum overnight to obtain the light black polystyrene microsphere loaded with the tetravalent platinum complex, namely the supported platinum complex oxidant.
The SEM results of the prepared supported platinum complex oxidant are shown in fig. 2.
The platinum loading of the supported platinum complex oxidant was determined by ICP-MS (inductively coupled plasma mass spectrometry) to be 0.25 mmol/g. By 2, 5-dimethoxythiophenol process (molecules, 2017, 22338.) determination of tetravalent platinum Complex of Supported platinum Complex oxidizing AgentsThe supported amount of (B) was 0.17 mmol/g.
Example 2
The polystyrene microspheres used in this example were commercially available and had particle sizes of 75 μm.
Adding 1.0g of polystyrene microspheres and 100mL of anhydrous pyridine into a 250 mL three-neck flask, then adding 5g of paratoluensulfonyl chloride, magnetically stirring, and reacting at room temperature for 2 d; after the reaction was completed, filtration was carried out, and the obtained solid was washed with ethanol 5 times and then 50 timesoC, vacuum drying for 12 hours to obtain yellow p-toluenesulfonyl modified polystyrene microspheres;
② adding 1.0g of p-toluenesulfonyl modified polystyrene microspheres into a 100mL three-necked flask, then adding 50mL of ethylenediamine, magnetically stirring, 60 percentoC, reacting for 1 d; after the reaction is finished, filtering, washing the solid with ethanol for 3 times, and then drying in vacuum to obtain dark yellow ethylenediamine modified polystyrene microspheres;
③ 0.2g of the ethylenediamine modified polystyrene microspheres were weighed into a 100mL three-necked flask, 30mL of DMF was added, and then 0.05g of cis-Pt (en) Cl was added2Ultrasonic dispersion, magnetic stirring, 90oC, reacting for 5 hours; after the reaction is finished, filtering and separating, washing the solid with DMF for 3 times, then washing with ethanol for 3 times, and then drying in vacuum overnight to obtain black polystyrene microspheres loaded with divalent platinum complexes;
and fourthly, weighing 0.1g of polystyrene microsphere loaded with the divalent platinum complex into a 100mL three-necked bottle, adding 50mL of ethanol, then adding 3g N-bromosuccinimide, magnetically stirring, reacting at room temperature for 12h, filtering and separating after the reaction is finished, washing the solid with ethanol for 3 times, and then drying in vacuum overnight to obtain the light black polystyrene microsphere loaded with the tetravalent platinum complex, namely the supported platinum complex oxidant.
Example 3
The polystyrene microspheres used in this example were commercially available and had particle sizes of 320 μm.
Adding 1.0g of polystyrene microspheres and 150 mL of anhydrous pyridine into a 250 mL three-neck flask, then adding 5g of paratoluensulfonyl chloride, magnetically stirring, and reacting at room temperature for 3 d; after the reaction is finished, the processFiltering, washing the obtained solid with ethanol for 5 times, and then 50 timesoC, vacuum drying for 12 hours to obtain yellow p-toluenesulfonyl modified polystyrene microspheres;
② adding 1.0g of p-toluenesulfonyl modified polystyrene microspheres into a 100mL three-necked flask, then adding 80mL of ethylenediamine, magnetically stirring, and 80 percentoC, reacting for 2 d; after the reaction is finished, filtering, washing the solid with ethanol for 3 times, and then drying in vacuum to obtain dark yellow ethylenediamine modified polystyrene microspheres;
③ 0.2g of the ethylenediamine modified polystyrene microspheres are weighed into a 100mL three-necked flask, 60mL of DMF is added, and then 0.1g of cis-Pt (en) Cl is added2Ultrasonic dispersion, magnetic stirring, 90oC, reacting for 5 hours; after the reaction is finished, filtering and separating, washing the solid with DMF for 3 times, then washing with ethanol for 3 times, and then drying in vacuum overnight to obtain black polystyrene microspheres loaded with divalent platinum complexes;
and fourthly, weighing 0.1g of polystyrene microsphere loaded with the divalent platinum complex into a 100mL three-necked bottle, adding 70mL of ethanol, then adding 5g N-bromosuccinimide, magnetically stirring, reacting at room temperature for 12h, filtering and separating after the reaction is finished, washing the solid with ethanol for 3 times, and then drying in vacuum overnight to obtain the light black polystyrene microsphere loaded with the tetravalent platinum complex, namely the supported platinum complex oxidant.
Example 4
The supported platinum complex oxidant (namely the polystyrene microsphere loaded with the tetravalent platinum complex) can react with various reduced polypeptides containing sulfydryl to synthesize polypeptide intramolecular disulfide bonds.
Synthesizing polypeptide intramolecular disulfide bonds by using polystyrene microspheres loaded with tetravalent platinum complexes and reduced polypeptides containing sulfydryl through the following scheme: carrying out oscillation reaction on the tetravalent platinum complex-loaded polystyrene microspheres and a reduced polypeptide aqueous solution for 0.5-1 h at room temperature, filtering and separating after the reaction is finished, and respectively recovering solids obtained by filtering and filtrate containing reaction products; separating the reaction product in the filtrate by using high performance liquid chromatography, and identifying the reaction product by mass spectrometry; and regenerating the polystyrene microspheres loaded with the tetravalent platinum complex after the regeneration treatment of the solid obtained by filtering, and recycling the polystyrene microspheres.
The oxidizing agent of the present invention can also oxidize various thiol-containing reduced polypeptides, such as reduced peptide (1), reduced RGD peptide, reduced oxytocin, reduced arginine vasopressin, reduced oxytocin. The reduced and oxidized amino acid sequences of these polypeptides are as follows:
specifically, the reduced polypeptide is oxidized by using the supported platinum complex oxidant prepared in example 1, and the specific reaction process is as follows:
firstly, reaction of supported platinum complex oxidant and reduced polypeptide (1)
In a 10mL reaction flask, 3mL of reduced polypeptide (1) solution with the concentration of 1mg/mL is prepared by water, then 15mg of supported platinum complex oxidant is added into the reaction flask, and the reaction is carried out for 0.5h at room temperature with shaking.
And (3) after the reaction is finished, filtering and separating the reaction system, filtering the obtained liquid phase, separating the product by using liquid chromatography, and measuring the molecular weight of the product by using mass spectrum: 1343.61, which corresponds to the signal peak of oxidized polypeptide (1) forming intramolecular disulfide bond; the yield is 99%; and (3) carrying out regeneration treatment on the solid phase obtained by filtering, specifically dispersing the solid phase into ethanol, and oxidizing the solid phase by using N-bromosuccinimide for 12 hours to complete regeneration (the specific regeneration operation process is the same as the condition of the step (r) in the example 1).
The regenerated oxidizing agent was reacted again with the same amount of reduced polypeptide (1), and thus, the experiment was repeated 5 times, and the yield of oxidized polypeptide (1) ranged from 98 to 99%.
FIG. 3 is a comparison of the chromatograms before and after the reaction of reduced polypeptide (1) with a supported platinum complex oxidant; as can be seen from the comparison of the figures: the reduced polypeptide (1) was completely oxidized to the oxidized polypeptide (1), and the oxidized polypeptide (1) was identified by mass spectrometry as shown in FIG. 4.
Secondly, reaction of load type platinum complex oxidant and reduction type RGD peptide
In a 10mL reaction bottle, 3mL of reduced RGD solution with the concentration of 1mg/mL is prepared by water, then 15mg of supported platinum complex oxidant is added into the reaction bottle, and the reaction is carried out for 0.5h at room temperature with shaking.
And (3) after the reaction is finished, filtering and separating the reaction system, filtering the obtained liquid phase, separating the product by using liquid chromatography, and measuring the molecular weight of the product by using mass spectrum: 946.38, which corresponds to the signal peak of oxidized RGD peptide forming intramolecular disulfide bond; and (3) regenerating the solid phase obtained by filtering, namely dispersing the solid phase into ethanol, and oxidizing the solid phase by using N-bromosuccinimide for 12 hours to complete regeneration.
The regenerated oxidant was reacted again with the same amount of reduced RGD peptide, and thus, the experiment was repeated 5 times, and the yield of oxidized RGD peptide ranged from 97 to 99%.
FIG. 5 is a comparison of chromatograms of reduced RGD peptide before and after reaction with a supported platinum complex oxidant; as can be seen from the comparison of the figures: the reduced RGD peptide was completely oxidized to oxidized RGD peptide, which was identified by mass spectrometry as shown in FIG. 6.
Thirdly, reaction of load type platinum complex oxidant and reduction type oxytocin
In a 5mL reaction bottle, 3mL of reduced oxytocin solution with the concentration of 1mg/mL is prepared by water, then 15mg of supported platinum complex oxidant is added into the reaction bottle, and the reaction is carried out for 0.5h under shaking at room temperature.
And (3) after the reaction is finished, filtering and separating the reaction system, filtering the obtained liquid phase, separating the product by using liquid chromatography, and measuring the molecular weight of the product by using mass spectrum: 1022.44, which corresponds to the signal peak of oxytocin forming intramolecular disulfide bonds; and (3) regenerating the solid phase obtained by filtering, namely dispersing the solid phase into ethanol, and oxidizing the solid phase by using N-bromosuccinimide for 12 hours to complete regeneration.
The regenerated oxidant was reacted again with the same amount of reduced oxytocin, and thus, the experiment was repeated 5 times, and the yield of oxidized RGD peptide ranged from 95 to 97%.
FIG. 7 is a comparison of chromatograms before and after reaction of reduced oxytocin with a supported platinum complex oxidant; as can be seen from the comparison of the figures: reduced oxytocin was completely oxidized to oxytocin, which was identified by mass spectrometry as shown in figure 8.
Fourthly, reaction of the supported platinum complex oxidant and reduced arginine vasopressin
In a 5mL reaction bottle, 3mL of reduced arginine vasopressin solution with the concentration of 1mg/mL is prepared by water, then 15mg of supported platinum complex oxidant is added into the reaction bottle, and the reaction is carried out for 0.5h under shaking at room temperature.
And (3) after the reaction is finished, filtering and separating the reaction system, filtering the obtained liquid phase, separating the product by using liquid chromatography, and measuring the molecular weight of the product by using mass spectrum: 1083.44, which corresponds to the signal peak of arginine vasopressin forming intramolecular disulfide bonds; and (3) regenerating the solid phase obtained by filtering, namely dispersing the solid phase into ethanol, and oxidizing the solid phase by using N-bromosuccinimide for 12 hours to complete regeneration.
The regenerated oxidant was reacted with the same amount of reduced arginine vasopressin again, and thus, the experiment was repeated 5 times with a yield of arginine vasopressin ranging from 96 to 97%.
FIG. 9 is a comparison of chromatograms of reduced arginine vasopressin before and after reaction with a supported platinum complex oxidant; as can be seen from the comparison of the figures: reduced arginine vasopressin was completely oxidized to arginine vasopressin, identified by mass spectrometry as shown in FIG. 10.
Fifthly, reacting the supported platinum complex oxidant with reduced oxytocin
In a 5mL reaction bottle, 3mL of reduced oxytocin solution with the concentration of 1mg/mL is prepared by water, then 15mg of supported platinum complex oxidant is added into the reaction bottle, and the reaction is carried out for 0.5h at room temperature with shaking.
And (3) after the reaction is finished, filtering and separating the reaction system, filtering the obtained liquid phase, separating the product by using liquid chromatography, and measuring the molecular weight of the product by using mass spectrum: 1005.44, which corresponds to the signal peak of oxytocin forming intramolecular disulfide bonds; and (3) regenerating the solid phase obtained by filtering, namely dispersing the solid phase into ethanol, and oxidizing the solid phase by using N-bromosuccinimide for 12 hours to complete regeneration.
The regenerated oxidant was reacted again with the same amount of reduced oxytocin, and thus, the experiment was repeated 5 times, and the yield of oxytocin ranged from 92 to 95%.
FIG. 11 is a comparison of chromatograms before and after reaction of reduced oxytocin with a supported platinum complex oxidant; as can be seen from the comparison of the figures: reduced oxytocin was completely oxidized to oxytocin, which was identified by mass spectrometry as shown in figure 12.
Comparative example 1
Adding acetonitrile into an acetic acid-sodium acetate buffer solution with the pH =4.65 according to the volume ratio of 1:1 to obtain a mixed solution, and introducing nitrogen into the mixed solution for 1h for later use; in a 20mL reaction flask, 8mL of the prepared mixed solution was prepared to have a concentration of 5.3X 10-4And (3) reacting the reduced polypeptide (1) solution with oxygen in the air in mol/L manner, and reacting for 22 hours by magnetic stirring to finish the reaction. The yield was 45%.
Comparative example 2
Introducing nitrogen into acetonitrile for 1h for later use; in a 20mL reaction flask, 8mL of a solution having a concentration of 5.3X 10 was prepared-4Adding 50 mu L of triethylamine into the reduced polypeptide (1) solution of mol/L, then adding 5mL of acetonitrile solution containing 10mg of iodine, reacting for 16h, removing excessive iodine simple substance after the reaction is finished, and separating the oxidized polypeptide (1) with the yield of 60%.
Comparative example 3
Adding acetonitrile into an acetic acid-sodium acetate buffer solution with the pH =4.65 according to the volume ratio of 1:1 to obtain a mixed solution, and introducing nitrogen into the mixed solution for 1h for later use; in a 20mL reaction flask, 8mL of the prepared mixed solution was prepared to have a concentration of 5.3X 10-4And (3) adding 0.1g of tetravalent platinum complex-loaded silica microspheres into the reaction flask, and oscillating for reaction at room temperature for 1h to isolate the oxidized polypeptide (1) with the yield of 68%.
The above examples and experimental data show that the supported platinum complex oxidant can be used for synthesis of polypeptide intramolecular disulfide bonds with different structures. Compared with the existing supported Ellman's oxidation reagent, the oxidant provided by the invention is simple to prepare and can be recycled, and the yield is not influenced by the recycling times. Compared with the silicon dioxide microsphere oxidant loaded with the tetravalent platinum complex, the yield is improved by 31 percent. Compared with the traditional oxidants such as iodine, oxygen and DMSO, the oxidant has the advantage of high yield, and when the oxidized polypeptide (1) is synthesized, the yield is only 45% by using oxygen as the oxidant; the yield was 60% using elemental iodine as the oxidant. After the polypeptide intramolecular disulfide bond is synthesized, the purification treatment of the polypeptide is simple, but the technical personnel in the field know that when DMSO is used as an oxidant, the purification of the polypeptide after reaction needs to be carried out multiple times of freeze drying treatment, and the DMSO is reduced to dimethyl sulfide with great toxicity, while the supported platinum complex oxidant of the invention takes water as a reaction solvent, and the purpose of separating products can be achieved by filtering after the reaction.
Claims (2)
1. The application of the supported platinum complex oxidant which is easy to recycle in preparation of polypeptide containing intramolecular disulfide bond is characterized in that the supported platinum complex oxidant is polystyrene microsphere loaded with tetravalent platinum complex, and has the structure shown in formula (I):
in the structure shown in formula (I), M is Pt4+Y is Br－The polystyrene microspheres are polyethylene glycol modified polystyrene microspheres, and the particle size of the polystyrene microspheres is 75-320 mu m; the supported platinum complex oxidant is prepared by the following method:
a) taking pyridine as a solvent, and reacting the polystyrene microspheres and p-toluenesulfonyl chloride for 2-3 d at room temperature according to the mass ratio of 1: 3-5;
after the reaction is finished, filtering, washing the obtained solid with ethanol, and then drying in vacuum to obtain a yellow solid;
b) ultrasonically dispersing the obtained yellow solid in ethylenediamine, and stirring and reacting for 1-2 d at the temperature of 60-80 ℃;
after the reaction is finished, cooling to room temperature, filtering, washing the obtained solid with ethanol, and drying in vacuum to obtain a dark yellow solid;
c) taking DMF as a solvent, and stirring and reacting the obtained dark yellow solid and a divalent platinum complex for 5-8 hours at the temperature of 70-90 ℃ according to the mass ratio of 1: 0.1-0.5; the divalent platinum complex is cis-Pt (en) Cl2；
After the reaction is finished, filtering and separating, washing the obtained solid, and then drying in vacuum to obtain a black solid;
d) ultrasonically dispersing the black solid in ethanol, and then fully reacting with N-bromosuccinimide at room temperature;
and after the reaction is finished, filtering and separating, washing the obtained solid, and drying in vacuum to obtain a light black solid, namely the supported platinum complex oxidant.
2. The use according to claim 1, wherein the polystyrene microspheres have a particle size of 110 μm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710934846.6A CN107892710B (en) | 2017-10-10 | 2017-10-10 | Supported platinum complex oxidant easy to recycle and use and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710934846.6A CN107892710B (en) | 2017-10-10 | 2017-10-10 | Supported platinum complex oxidant easy to recycle and use and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107892710A CN107892710A (en) | 2018-04-10 |
| CN107892710B true CN107892710B (en) | 2021-04-13 |
Family
ID=61803538
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710934846.6A Expired - Fee Related CN107892710B (en) | 2017-10-10 | 2017-10-10 | Supported platinum complex oxidant easy to recycle and use and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107892710B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109912477B (en) * | 2019-03-07 | 2020-12-29 | 河北大学 | Selenomethionine oxide, preparation method thereof and application thereof in synthesis of polypeptide intramolecular disulfide bond |
| CN109836470B (en) * | 2019-03-22 | 2022-03-22 | 河北大学 | Method for automatically synthesizing polypeptide intramolecular disulfide bond |
| WO2022235852A1 (en) * | 2021-05-04 | 2022-11-10 | Cend Therapeutics, Inc. | Irgd-analogs and related therapeutic methods |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105085603A (en) * | 2015-08-26 | 2015-11-25 | 河北大学 | Reusable load type platinum complex oxidizing agents, and preparation method and application thereof |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6365592B1 (en) * | 1999-05-14 | 2002-04-02 | Merck Frosst Canada & Co. | Phosphonic and carboxylic acid derivatives as inhibitors of protein tyrosine phosphatase-1B (PTP-1B) |
| CN101490052B (en) * | 2006-05-12 | 2012-08-08 | 美瑞德生物工程公司 | Therapeutic compounds and their use in cancer |
| EP2451461A4 (en) * | 2009-07-06 | 2013-05-29 | Ontorii Inc | Novel nucleic acid prodrugs and methods of use thereof |
| EP2493910A1 (en) * | 2009-10-30 | 2012-09-05 | Tranzyme Pharma, Inc. | Macrocyclic ghrelin receptor antagonists and inverse agonists and methods of using the same |
| CN106565884A (en) * | 2016-10-19 | 2017-04-19 | 黑龙江大学 | Preparing method for polystyrene microsphere loaded nano-copper composite material |
| CN106866744B (en) * | 2017-02-20 | 2018-11-13 | 河北大学 | A kind of tetravalence platinum complex oxidant and its preparation method and application |
-
2017
- 2017-10-10 CN CN201710934846.6A patent/CN107892710B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105085603A (en) * | 2015-08-26 | 2015-11-25 | 河北大学 | Reusable load type platinum complex oxidizing agents, and preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| "Mono- and di-bromo platinum(IV) prodrugs via oxidative bromination: synthesis, characterization, and cytotoxicity";Zoufeng Xu 等;《Dalton Trans》;20151214;第44卷(第46期);摘要 * |
| "Pt(Ⅳ)配合物和负载型 Pt(Ⅳ)配合物氧化合成多肽分子内二硫键的研究";张亚梅;《中国优秀硕士学位论文全文数据库（电子期刊） 工程科技I辑》;20190115(第01期);第B014-1026页 * |
| "负载铂配合物的聚合物微球合成催产素分子内二硫键的研究";刘蒙雨;《中国优秀硕士学位论文全文数据库（电子期刊） 工程科技I辑》;20170315(第03期);摘要，正文第6页第1段，第8-10页，第14页第1段，图2-1 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107892710A (en) | 2018-04-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP6554180B6 (en) | Solid phase synthesis method of DNA-encoded compound library | |
| CN107892710B (en) | Supported platinum complex oxidant easy to recycle and use and preparation method and application thereof | |
| CN105085603B (en) | A kind of load type platinum complex oxidant that can be recycled and its preparation method and application | |
| CN115028849B (en) | Preparation method and application of metal triangle supermolecule with room temperature fluorescence | |
| CN105964306B (en) | It is a kind of based on poly ion liquid magnetic nano-particle, preparation method and its application in three component reactions | |
| CN106866744B (en) | A kind of tetravalence platinum complex oxidant and its preparation method and application | |
| Zhao et al. | Cooperative chiral salen Ti IV catalysts with built-in phase-transfer capability accelerate asymmetric sulfoxidation in water | |
| WO2022050236A1 (en) | Production method for alkaline earth metal formate | |
| CN106905254B (en) | A kind of preparation method of 5- phenyl -1H- tetrazole | |
| CN102391288B (en) | Preparation methods of cefpirome intermediate and cefpirome | |
| CN108864084B (en) | Apixaban related substances and preparation method thereof | |
| CN109232586A (en) | A kind of synthetic method of the more imidazole salts cage compounds of three-dimensional | |
| CN114874227A (en) | Catalyst for amide synthesis and preparation and application thereof | |
| WO2021184649A1 (en) | Method for preparing florfenicol intermediate and compound obtained by method | |
| CN110183513B (en) | Cyclopeptide compound simulating structure of natural product and preparation method thereof | |
| JP7405594B2 (en) | Method for producing formic acid | |
| CN110156641B (en) | Synthesis method of substituted guanidine | |
| CN110041274B (en) | Method for preparing 5-fluoroalkyl triazole compound by air oxidation multi-component one-pot method | |
| JP3834647B2 (en) | Method for producing arylanthracene compound | |
| CN115784933B (en) | Difunctional nitrile oxide, preparation method and application thereof | |
| CN109912477B (en) | Selenomethionine oxide, preparation method thereof and application thereof in synthesis of polypeptide intramolecular disulfide bond | |
| CN111378009A (en) | Preparation method of octreotide | |
| CN113956268B (en) | 6-bromo-1-chlorobenzothiophene [2,3-c ] pyridine and synthetic method | |
| CN116554044A (en) | 2, 6-diisopropyl-4-carboxyaniline without amino protection and synthesis method thereof | |
| JP7309414B2 (en) | SUZUKI-MIYAURA COUPLING REACTION CATALYST AND METHOD FOR PRODUCING SAME |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20210413 Termination date: 20211010 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |