CN107892710A - A kind of load type platinum complex oxidant for being easily recycled recycling and its preparation method and application - Google Patents
A kind of load type platinum complex oxidant for being easily recycled recycling and its preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of load type platinum complex oxidant for being easily recycled recycling.Load type platinum complex oxidant has such as formula（Ⅰ）Shown structure.Present invention also offers the preparation method of the oxidant：a）Using pyridine as solvent, polystyrene microsphere and paratoluensulfonyl chloride are reacted to obtain yellow solid；b）By gained yellow solid, simultaneously stirring reaction obtains dark yellow solid thing to ultrasonic disperse in ethylenediamine；c）Using DMF as solvent, gained dark yellow solid thing and divalence platinum complex stirring reaction are obtained into black solid thing；d）By black solid thing in ethanol after ultrasonic disperse with N, the light/dark balance solids that the bromo-succinimides of N ' two fully react to obtain is load type platinum complex oxidant.The preparation method of the oxidant of the present invention is simple, easily recycling and reaction yield height.
Description
Technical field
The present invention relates to load type platinum complex oxidant, specifically a kind of support type for being easily recycled recycling
Platinum complex oxidant and its preparation method and application.
Background technology
Disulfide bond is to maintain polypeptide higher order conformation in peptide molecule, improves the important official of polypeptide physiologically active and pharmacological activity
Can group.Disulfide bond in pharmaceutical polypeptide molecule can improve stability, the permeability of drug molecule in vivo, increase drug molecule
With the specific binding of target molecules, so as to improve the bioactivity of polypeptide drugs（Nature Chem., 2014, 6,
1009–1016; Chem. Rew., 2013, 114, 901–926.）.Many polypeptide drugs containing intramolecular disulfide bond by
FDA ratifies and is used to treat various diseases, such as：Treat the Octreotide of various endocrine knurls, treat the Nai Xili of heart failure
Peptide, calcitonin, uterotonics oxytocins, antiplatelet drug eptifibatide, the Remedies for diabetes for treating Paget ' s diseases
The polypeptide drugs such as pramlintide.At present, largely the polypeptide containing intramolecular disulfide bond is developed, and promotes polypeptide drug
Development（Future Med. Chem., 2009, 1, 361–377.）.The generation of polypeptide drug intramolecular disulfide bond is essence
An important reaction in thin organic chemical industry and pharmaceutical chemistry.
The method of traditional synthesis polypeptide intramolecular disulfide bond is：（I）Reacted by solid-liquid, on solid phase carrier directly
Form disulfide bond in peptide molecule.Gu the reaction between-liquid is carried out under quasi- diluting condition, the shape of intramolecular disulfide bond is promoted
Into reducing the formation of disulfide bond between accessory substance peptide molecule.But the yield of the polypeptide containing intramolecular disulfide bond is by solid phase carrier
Load capacity has a great influence, and yield is undesirable.（II）By homogeneous reaction, oxidative folding forms intramolecular disulfide bond in the liquid phase
（Eur. J. Org. Chem., 2014, 3519–3530.）.
Being presently used for the oxidant that polypeptide drug intramolecular disulfide is bonded to includes：Iodine, thallium trifluoroacetate (III), oxygen
Gas, DMSO, oxidized form of glutathione and Ellman ' s reagents etc..But these oxidants are in synthesis polypeptide intramolecular disulfide bond
When, with the methionine in polypeptide chain, tyrosine, trp residue side reaction occurs for some meetings, and some meetings make polypeptide generate dimerization
The accessory substance of body or polymer, causes yield to reduce, and oxidant can not recycle, some oxidants can produce toxicity compared with
Big accessory substance (Eur. J. Org. Chem., 2014, 3519–3530; RSC Adv. 2014, 4, 13854–
13881; Synthesis, 2008, 16, 2491–2509.) 。
In addition to above-mentioned conventional oxidant, tetravalence platinum complex oxidant obtains extensively in synthetic drug intramolecular disulfide bond
General application, and oxidation effectiveness is superior to presently used other oxidants（J. Am. Chem. Soc. 2000, 122,
6809–6815; J. Org. Chem. 1999, 64, 4590–4595; Bioorg.Med. Chem. Lett. 2002,12, 2237–2240.）.But platinum complex due to it is expensive, the reason such as be not easy to prepare and limit its application, therefore, such as
Fruit can prepare a kind of platinum complex oxidant for being easily recycled and using, then can substantially reduce and carry out synthetic drug using such oxidant
The cost payout of intramolecular disulfide bond.
The content of the invention
An object of the present invention is to provide a kind of load type platinum complex oxidant for being easily recycled recycling and should
With so that solve can not recycling or reclaimer operation when existing platinum complex oxidant is used for synthesis polypeptide intramolecular disulfide bond
The problem of more complicated.
The second object of the present invention is to provide a kind of system for the load type platinum complex oxidant for being easily recycled recycling
Preparation Method.
What an object of the present invention was realized in：
A kind of load type platinum complex oxidant for being easily recycled recycling, it is the polystyrene of load tetravalence platinum complex
Microballoon, it has such as formula（Ⅰ）Shown structure：
（Ⅰ）
Formula（Ⅰ）In shown structure, M Pt4+, Y Br-, the particle diameter of polystyrene microsphere is 75 μm ~ 320 μm.
The polystyrene microsphere is polyethyleneglycol modified polystyrene microsphere.
The particle diameter of the polystyrene microsphere is 110 μm.
Present invention also offers the load type platinum complex oxidant in the polypeptide containing intramolecular disulfide bond is prepared
Using.
The application can be specifically achieved through the following technical solutions：
By load type platinum complex oxidant and reduced form polypeptide solution, the h of concussion reaction 0.5 ~ 1, reaction are tied at ambient temperature
Shu Hou, it is separated by filtration, filtering gained solids and the filtrate containing reaction product is separately recovered；Utilize high performance liquid chromatography separation
Reaction product in filtrate；Load type platinum complex oxidant, circulation profit are regenerated after filtering gained solids regeneration treatment
With.
The load type platinum complex oxidant is with the reduced form polypeptide amount ratio with tetravalence platinum complex and reduced form
Polypeptide mol ratio is advisable for 1 ~ 1.2: 1.
The reduced form polypeptide solution is made of using reduced form polypeptide as solute, using water as solvent.
The reduced form polypeptide is reduced form polypeptide containing sulfydryl, can to form intramolecular disulfide bond, is specifically included
Reduced form polypeptide (1), reduced form RGD peptide, reduced form oxypressin, reduced form arginine vasopressin and reduced form oxytocins.
The amino acid sequence of the reduced form peptide (1) is：H-Cys-Gly-Tyr-Cys-His-Lys-Leu-His-Gln-Met-NH2；
The amino acid sequence of the reduced form RGD peptide is：H-Cys-Arg-Gly-Asp-Lys-Gly-Pro-Asp-Cys-NH2；It is described
The amino acid sequence of reduced form oxypressin is：H-Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Lys-Gly-NH2；It is described
The amino acid sequence of reduced form arginine vasopressin is：H-Cys-Tyr-Phe-Gln-Asn-Cys-Pro-Arg-Gly-NH2；Institute
The amino acid sequence for stating reduced form oxytocins is：H-Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly-NH2。
What the second object of the present invention was realized in：
A kind of preparation method for the load type platinum complex oxidant for being easily recycled recycling, comprises the following steps：
a）Using pyridine as solvent, polystyrene microsphere and paratoluensulfonyl chloride in mass ratio 1: 3 ~ 5 react at ambient temperature
2~3d；
Reaction is finished, and filtering, gained solid is dried in vacuo after being washed with ethanol, obtains yellow solid；
b）By gained yellow solid in ethylenediamine ultrasonic disperse, 1 ~ 2d of stirring reaction under the conditions of 60 ~ 80 DEG C；
Reaction is finished, and is filtered after being cooled to room temperature, and gained solid is dried in vacuo after being washed with ethanol, obtains dark yellow solid thing；
c）Using DMF as solvent, by gained dark yellow solid thing and divalence platinum complex in mass ratio 1: 0.1 ~ 0.5, at 70 ~ 90 DEG C
Under the conditions of 5 ~ 8h of stirring reaction；
Reaction finishes, and is separated by filtration, and is dried in vacuo after the washing of gained solids, obtains black solid thing；
d）Black solid thing after ultrasonic disperse, is fully reacted with N, the bromo-succinimides of N '-two at room temperature in ethanol；
Reaction finishes, and is separated by filtration, and is dried in vacuo after the washing of gained solids, obtained light/dark balance solids is load type platinum
Complex oxidant.
The preparation method of load type platinum complex oxidant of the present invention, step（a）In, the polystyrene microsphere is
Polyethyleneglycol modified polystyrene microsphere.The particle diameter of polystyrene microsphere is 75 μm ~ 320 μm.
The preparation method of load type platinum complex oxidant of the present invention, step（c）In, the divalence platinum complex is
Cis Pt (en) Cl2。
The preparation method of load type platinum complex oxidant of the present invention, step（d）In, in mol, the N, N '-
The dosage of two bromo-succinimides is not less than 2 times of the divalence platinum complex dosage.
The oxidant of the present invention and its preparation have many advantages：
1st, load type platinum complex oxidant of the present invention is prepared simple, and can be recycled after regeneration treatment, synthesized
The yield of oxidized form polypeptide is not recycled number by this oxidant to be influenceed.
2nd, there is the advantages of yield is high relative to existing oxidant, load type platinum complex oxidant of the present invention.
3rd, after synthesis polypeptide intramolecular disulfide bond, the recovery and regeneration treatment of oxidant are simple, and load type platinum of the present invention is matched somebody with somebody
Compound oxidant can reach the purpose of separation product using water as reaction dissolvent by filtering after reaction.
Brief description of the drawings
Fig. 1 is that load type platinum complex oxidant of the present invention prepares reacting flow chart.
Fig. 2 is the SEM scanning figures of load type platinum complex oxidant of the present invention.
Fig. 3 is reduced form polypeptide（1）Scheme with the Chromatographic Comparison before and after load type platinum complex oxidant reaction of the present invention.
Fig. 4 is oxidized form polypeptide（1）Mass spectrogram.
Fig. 5 is that reduced form RGD peptide is schemed with the Chromatographic Comparison before and after load type platinum complex oxidant reaction of the present invention.
Fig. 6 is the mass spectrogram of oxidized form RGD peptide.
Fig. 7 is reduced form oxypressin and the Chromatographic Comparison before and after load type platinum complex oxidant reaction of the present invention
Figure.
Fig. 8 is the mass spectrogram of oxidized form oxypressin.
Fig. 9 is reduced form arginine vasopressin and the Chromatographic Comparison before and after load type platinum complex oxidant reaction of the present invention
Figure.
Figure 10 is the mass spectrogram of oxidized form arginine vasopressin.
Figure 11 is that reduced form oxytocins is schemed with the Chromatographic Comparison before and after load type platinum complex oxidant reaction of the present invention.
Figure 12 is the mass spectrogram of oxidized form oxytocins.
Embodiment
What Fig. 1 gave load type platinum complex oxidant of the present invention prepares reacting flow chart.It is as shown in figure 1, of the invention
Polyethyleneglycol modified polystyrene microsphere and paratoluensulfonyl chloride are reacted, then and ethylenediamine（Logical nitrogen）Reaction, obtains second
Diamine modified polystyrene microsphere（For dark yellow solid thing）, ethylene diamine-modified polystyrene microsphere and cis dichloride second
Diamines closes platinum（Cis Pt（en）Cl2）Reaction, obtain loading the polystyrene microsphere of divalence platinum complex（Black solid thing）, bear
Oxidation reaction occurs for the polystyrene microsphere and N, N '-two bromo-succinimide for carrying divalence platinum complex, obtains loading tetravalence
The polystyrene microsphere of platinum complex（Light/dark balance solids）, load type platinum complex oxidant as of the present invention, its specific system
Standby process is shown in embodiment 1 ~ 3：
Embodiment 1
Polystyrene microsphere used in the present embodiment obtains to be purchased in market, 110 μm of particle diameter.
1. in 250 mL three-neck flasks, 1.0 g polystyrene microspheres and 100 mL anhydrous pyridines are added, then add 5
G paratoluensulfonyl chlorides, magnetic agitation, react at room temperature 3 d；Reaction finishes, and filtering, gained solid washs 5 times with ethanol, then
50 oC is dried in vacuo 12h, obtains the polystyrene microsphere of the p-toluenesulfonyl modification of yellow；
2. the polystyrene microsphere that 1.0g p-toluenesulfonyls are modified is added in 100mL three-necked bottle, 50mL is then added
Ethylenediamine, magnetic agitation, 60oC reacts 2d；Reaction is finished, and filtering, solid is washed 3 times with ethanol, is then dried in vacuo, obtains
The ethylene diamine-modified polystyrene microsphere of buff；
3. weighing the ethylene diamine-modified polystyrene microspheres of 0.2g in 100mL three-necked bottle, 30mL DMF, Ran Houjia are added
Enter 0.1g cis Pt (en) Cl2, ultrasonic disperse, magnetic agitation, 90oC reacts 5h；After completion of the reaction, it is separated by filtration, solid is used
DMF is washed 3 times, then is washed 3 times with ethanol, is then dried in vacuum overnight, is obtained the polyphenyl of the load divalence platinum complex of black
Ethene microballoon；
4. weighing the polystyrene microsphere of 0.1g load divalence platinum complexes in 100mL three-necked bottle, 50mL ethanol is added, so
After add 5g N, the bromo-succinimides of N '-two, magnetic agitation, react 12h at room temperature, after completion of the reaction, be separated by filtration,
Solid is washed 3 times with ethanol, is then dried in vacuum overnight, and the polystyrene for obtaining the load tetravalence platinum complex of light/dark balance is micro-
Ball, i.e., load type platinum complex oxidant of the present invention.
The SEM results of prepared load type platinum complex oxidant are as shown in Figure 2.
ICP-MS（Inductivity coupled plasma mass spectrometry）The load capacity of platinum for determining load type platinum complex oxidant is
0.25mmol/g.Utilize 2,5- dimethoxy benzenethiol methods（molecules, 2017, 22, 338.）Determine load type platinum
The load capacity of the tetravalence platinum complex of complex oxidant is 0.17mmol/g.
Embodiment 2
Polystyrene microsphere used in the present embodiment obtains to be purchased in market, 75 μm of particle diameter.
1. in 250 mL three-neck flasks, 1.0 g polystyrene microspheres and 100 mL anhydrous pyridines are added, then add 5
G paratoluensulfonyl chlorides, magnetic agitation, react at room temperature 2d；Reaction finishes, and filtering, gained solid washs 5 times with ethanol, and then 50oC is dried in vacuo 12h, obtains the polystyrene microsphere of the p-toluenesulfonyl modification of yellow；
2. the polystyrene microsphere that 1.0g p-toluenesulfonyls are modified is added in 100mL three-necked bottle, 50mL is then added
Ethylenediamine, magnetic agitation, 60oC reacts 1d；Reaction is finished, and filtering, solid is washed 3 times with ethanol, is then dried in vacuo, obtains
The ethylene diamine-modified polystyrene microsphere of buff；
3. weighing the ethylene diamine-modified polystyrene microspheres of 0.2g in 100mL three-necked bottle, 30mL DMF, Ran Houjia are added
Enter 0.05g cis Pt (en) Cl2, ultrasonic disperse, magnetic agitation, 90oC reacts 5h；After completion of the reaction, it is separated by filtration, solid is used
DMF is washed 3 times, then is washed 3 times with ethanol, is then dried in vacuum overnight, is obtained the polyphenyl of the load divalence platinum complex of black
Ethene microballoon；
4. weighing the polystyrene microsphere of 0.1g load divalence platinum complexes in 100mL three-necked bottle, 50mL ethanol is added, so
After add 3g N, the bromo-succinimides of N '-two, magnetic agitation, react 12h at room temperature, after completion of the reaction, be separated by filtration,
Solid is washed 3 times with ethanol, is then dried in vacuum overnight, and the polystyrene for obtaining the load tetravalence platinum complex of light/dark balance is micro-
Ball, i.e., load type platinum complex oxidant of the present invention.
Embodiment 3
Polystyrene microsphere used in the present embodiment obtains to be purchased in market, 320 μm of particle diameter.
1. in 250 mL three-neck flasks, 1.0 g polystyrene microspheres and 150 mL anhydrous pyridines are added, then add 5
G paratoluensulfonyl chlorides, magnetic agitation, react at room temperature 3d；Reaction finishes, and filtering, gained solid washs 5 times with ethanol, and then 50oC is dried in vacuo 12h, obtains the polystyrene microsphere of the p-toluenesulfonyl modification of yellow；
2. the polystyrene microsphere that 1.0g p-toluenesulfonyls are modified is added in 100mL three-necked bottle, 80mL is then added
Ethylenediamine, magnetic agitation, 80oC reacts 2d；Reaction is finished, and filtering, solid is washed 3 times with ethanol, is then dried in vacuo, obtains
The ethylene diamine-modified polystyrene microsphere of buff；
3. weighing the ethylene diamine-modified polystyrene microspheres of 0.2g in 100mL three-necked bottle, 60mL DMF, Ran Houjia are added
Enter 0.1g cis Pt (en) Cl2, ultrasonic disperse, magnetic agitation, 90oC reacts 5h；After completion of the reaction, it is separated by filtration, solid is used
DMF is washed 3 times, then is washed 3 times with ethanol, is then dried in vacuum overnight, is obtained the polyphenyl of the load divalence platinum complex of black
Ethene microballoon；
4. weighing the polystyrene microsphere of 0.1g load divalence platinum complexes in 100mL three-necked bottle, 70mL ethanol is added, so
After add 5g N, the bromo-succinimides of N '-two, magnetic agitation, react 12h at room temperature, after completion of the reaction, be separated by filtration,
Solid is washed 3 times with ethanol, is then dried in vacuum overnight, and the polystyrene for obtaining the load tetravalence platinum complex of light/dark balance is micro-
Ball, i.e., load type platinum complex oxidant of the present invention.
Embodiment 4
The load type platinum complex oxidant of the present invention（Load the polystyrene microsphere of tetravalence platinum complex）It can contain with a variety of
There is the reduced form polypeptides reactive synthesis polypeptide intramolecular disulfide bond of sulfydryl.
The polystyrene microsphere of load tetravalence platinum complex is synthesized with the reduced form polypeptide containing sulfydryl by following scheme
Disulfide bond in peptide molecule：By the polystyrene microsphere of load tetravalence platinum complex with reduced form polypeptid solution in room temperature condition
0.5 ~ 1h of lower concussion reaction, after reaction terminates, it is separated by filtration, filtering gained solids and the filter containing reaction product is separately recovered
Liquid；Reaction product is identified using the reaction product in high performance liquid chromatography separation filtrate, and by mass spectrum；Filtering gained
The polystyrene microsphere of load tetravalence platinum complex is regenerated after solids regeneration treatment, is recycled.
The also oxidable a variety of reduced form polypeptides containing sulfydryl of oxidant of the present invention, such as reduced form peptide (1), reduced form
RGD peptide, reduced form oxypressin, reduced form arginine vasopressin, reduced form oxytocins.The reduced form of these polypeptides and oxidation
Type amino acid sequence is as follows：
Specifically, utilize the oxidizing above-mentioned reduced form polypeptide of the load type platinum complex prepared by embodiment 1, specific reaction
Process is as follows：
First, load type platinum complex oxidant and reduced form polypeptide（1）Reaction
In 10mL reaction bulb, the reduced form polypeptide that 3mL concentration is 1mg/mL is prepared with water（1）Solution, then to reaction bulb
Middle addition 15mg load type platinum complex oxidants, room temperature concussion reaction 0.5h.
Reaction terminates, and reaction system is separated by filtration, and filtering gained liquid phase liquid chromatogram separation product, uses mass spectroscopy
The molecular weight of product is：1343.61, the result meets the oxidized form polypeptide to form intramolecular disulfide bond（1）Signal peak；Yield
For 99%；Filtering gained solid phase carries out regeneration treatment, and specific method is by solid phase dispersion into ethanol, with N, the bromos of N '-two fourth two
Acid imide oxidation 12h completes regeneration（4. condition is identical with step in embodiment 1 for specific non-regenerative operating procedure）.
By the oxidant after regeneration again with same amount of reduced form polypeptide（1）Reaction, in this way, repeating experiment 5 times, oxidized form
Polypeptide（1）Yield scope be 98-99%.
Fig. 3 is reduced form polypeptide（1）Scheme with Chromatographic Comparison before and after load type platinum complex oxidant reaction；Being compared by figure can
Know：Reduced form polypeptide（1）It is fully oxidized as oxidized form polypeptide（1）, oxidized form polypeptide（1）It is as shown in Figure 4 with Mass Spectrometric Identification.
2nd, load type platinum complex oxidant and reduced form RGD peptide react
In 10mL reaction bulb, the reduced form RGD solution that 3mL concentration is 1mg/mL is prepared with water, is then added into reaction bulb
Enter 15mg load type platinum complex oxidants, room temperature concussion reaction 0.5h.
Reaction terminates, and reaction system is separated by filtration, and filtering gained liquid phase liquid chromatogram separation product, uses mass spectroscopy
The molecular weight of product is：946.38, the result meets the signal peak for the oxidized form RGD peptide to form intramolecular disulfide bond；Filtering institute
Obtain solid phase and carry out regeneration treatment, specific method is by solid phase dispersion into ethanol, with N, the oxidation of the bromo-succinimides of N '-two
12h completes regeneration.
Oxidant after regeneration is reacted with same amount of reduced form RGD peptide again, in this way, repeating experiment 5 times, oxidized form
The yield scope of RGD peptide is 97-99%.
Fig. 5 is that reduced form RGD peptide is schemed with the Chromatographic Comparison before and after load type platinum complex oxidant reaction；Being compared by figure can
Know：Reduced form RGD peptide be fully oxidized for oxidized form RGD peptide, oxidized form RGD peptide Mass Spectrometric Identification it is as shown in Figure 6.
3rd, load type platinum complex oxidant and reduced form oxypressin react
In 5mL reaction bulb, the reduced form oxypressin solution that 3mL concentration is 1mg/mL is prepared with water, then to reaction
15mg load type platinum complex oxidants, room temperature concussion reaction 0.5h are added in bottle.
Reaction terminates, and reaction system is separated by filtration, and filtering gained liquid phase liquid chromatogram separation product, uses mass spectroscopy
The molecular weight of product is：1022.44, the result meets the signal peak for the oxypressin to form intramolecular disulfide bond；Filtering institute
Obtain solid phase and carry out regeneration treatment, specific method is by solid phase dispersion into ethanol, with N, the oxidation of the bromo-succinimides of N '-two
12h completes regeneration.
Oxidant after regeneration is reacted with same amount of reduced form oxypressin again, in this way, repeating experiment 5 times, oxygen
The yield scope of change type RGD peptide is 95-97%.
Fig. 7 is that reduced form oxypressin is schemed with the Chromatographic Comparison before and after load type platinum complex oxidant reaction；By scheming
Compare and understand：Reduced form oxypressin is fully oxidized as oxypressin, oxypressin Mass Spectrometric Identification such as Fig. 8 institutes
Show.
4th, load type platinum complex oxidant and reduced form arginine vasopressin react
In 5mL reaction bulb, the reduced form arginine vasopressin solution that 3mL concentration is 1mg/mL is prepared with water, then to anti-
Answer addition 15mg load type platinum complex oxidants, room temperature concussion reaction 0.5h in bottle.
Reaction terminates, and reaction system is separated by filtration, and filtering gained liquid phase liquid chromatogram separation product, uses mass spectroscopy
The molecular weight of product is：1083.44, the result meets the signal peak for the arginine vasopressin to form intramolecular disulfide bond；Filtering
Gained solid phase carries out regeneration treatment, and specific method is by solid phase dispersion into ethanol, with N, the oxidation of the bromo-succinimides of N '-two
12h completes regeneration.
Oxidant after regeneration is reacted with same amount of reduced form arginine vasopressin again, in this way, experiment 5 times is repeated,
The yield scope of arginine vasopressin is 96-97%.
Fig. 9 is that reduced form arginine vasopressin is schemed with the Chromatographic Comparison before and after load type platinum complex oxidant reaction；By
Figure is relatively understood：Reduced form arginine vasopressin is fully oxidized as arginine vasopressin, arginine vasopressin Mass Spectrometric Identification
As shown in Figure 10.
5th, load type platinum complex oxidant and reduced form oxytocins react
In 5mL reaction bulb, the reduced form oxytocins solution that 3mL concentration is 1mg/mL is prepared with water, then into reaction bulb
Add 15mg load type platinum complex oxidants, room temperature concussion reaction 0.5h.
Reaction terminates, and reaction system is separated by filtration, and filtering gained liquid phase liquid chromatogram separation product, uses mass spectroscopy
The molecular weight of product is：1005.44, the result meets the signal peak for the oxytocins to form intramolecular disulfide bond；Filtering gained is solid
Regeneration treatment is mutually carried out, specific method is by solid phase dispersion into ethanol, and with N, it is complete that the bromo-succinimides of N '-two aoxidize 12h
Into regeneration.
Oxidant after regeneration is reacted with same amount of reduced form oxytocins again, in this way, repeating experiment 5 times, oxytocins
Yield scope be 92-95%.
Figure 11 is that reduced form oxytocins is schemed with the Chromatographic Comparison before and after load type platinum complex oxidant reaction；Compared by figure
Understand：Reduced form oxytocins be fully oxidized for oxytocins, oxytocins Mass Spectrometric Identification it is as shown in figure 12.
Comparative example 1
The NaAc_HAc buffer solution of pH=4.65 by volume 1:1 ratio adds acetonitrile and obtains mixed solution, and mixing is molten
It is standby that liquid leads to nitrogen 1h；In 20mL reaction bulb, it is 5.3 × 10 to prepare 8mL concentration with the mixed solution of preparation-4Mol/L's
Reduced form polypeptide（1）Solution, this solution and the oxygen reaction in air, magnetic agitation reaction 22h, reaction terminate.Yield is
45%。
Comparative example 2
It is standby that acetonitrile leads to nitrogen 1h；In 20mL reaction bulb, it is 5.3 × 10 to prepare 8mL concentration-4Mol/L reduced form polypeptide
（1）Solution, 50 μ L triethylamines are added, then add the acetonitrile solution that 5mL contains 10mg iodine, react 16h, reaction terminates, and removes
Excessive elemental iodine, separate oxidized form polypeptide（1）, yield 60%.
Comparative example 3
The NaAc_HAc buffer solution of pH=4.65 by volume 1:1 ratio adds acetonitrile and obtains mixed solution, and mixing is molten
It is standby that liquid leads to nitrogen 1h；In 20mL reaction bulb, it is 5.3 × 10 to prepare 8mL concentration with the mixed solution of preparation-4Mol/L's
Reduced form polypeptide（1）Solution, the silicon dioxide microsphere of 0.1g load tetravalence platinum complexes, room temperature shake are then added into reaction bulb
Reaction 1h is swung, separates oxidized form polypeptide（1）, yield 68%.
Above example and experimental data show that load type platinum complex oxidant of the invention can be used in different structure
Peptide molecule in disulfide bond synthesis.The oxidant of the present invention is compared with existing support type Ellman ' s oxidising agents, this hair
Preparing for bright oxidant is simple, and can be recycled, and yield is not influenceed by number is recycled.With loading tetravalence platinum complex
Silicon dioxide microsphere oxidant compared to yield improve 31%.Compared to traditional oxidant such as iodine, oxygen, DMSO, oxygen of the present invention
Agent has the advantages of yield is high, when synthesizing oxidized form polypeptide (1), there was only 45% by the use of oxygen as oxidant, yield；Profit
By the use of elemental iodine as yield during oxidant be 60%.After synthesis polypeptide intramolecular disulfide bond of the present invention, the purification processes letter of polypeptide
It is single, and it is as well known to those skilled in the art when by the use of DMSO as oxidant, and the purification needs of polypeptide are repeatedly freezed after reaction
Drying process, and DMSO is reduced to the very big dimethyl sulfide of toxicity, and load type platinum complex oxidant of the present invention is with water
For reaction dissolvent, pass through the purpose of the i.e. reachable separation product of filtering after reaction.
Claims (8)
1. a kind of load type platinum complex oxidant for being easily recycled recycling, it is characterized in that, it coordinates for load tetravalence platinum
The polystyrene microsphere of thing, it has such as formula（Ⅰ）Shown structure：
（Ⅰ）
Formula（Ⅰ）In shown structure, M Pt4+, Y Br-, the particle diameter of polystyrene microsphere is 75 μm ~ 320 μm.
2. the load type platinum complex oxidant of recycling is easily recycled according to claim 1, it is characterized in that, it is described poly-
Phenylethylene micro ball is polyethyleneglycol modified polystyrene microsphere.
3. the load type platinum complex oxidant of recycling is easily recycled according to claim 1, it is characterized in that, it is described poly-
The particle diameter of phenylethylene micro ball is 110 μm.
4. claim 1 ~ 3 any load type platinum complex oxidant answering in the polypeptide containing intramolecular disulfide bond is prepared
With.
5. a kind of preparation method for the load type platinum complex oxidant for being easily recycled recycling, it is characterized in that, including it is following
Step：
a）Using pyridine as solvent, polystyrene microsphere and paratoluensulfonyl chloride in mass ratio 1: 3 ~ 5 react at ambient temperature
2~3d；
Reaction is finished, and filtering, gained solid is dried in vacuo after being washed with ethanol, obtains yellow solid；
b）By gained yellow solid in ethylenediamine ultrasonic disperse, 1 ~ 2d of stirring reaction under the conditions of 60 ~ 80 DEG C；
Reaction is finished, and is filtered after being cooled to room temperature, and gained solid is dried in vacuo after being washed with ethanol, obtains dark yellow solid thing；
c）Using DMF as solvent, by gained dark yellow solid thing and divalence platinum complex in mass ratio 1: 0.1 ~ 0.5, at 70 ~ 90 DEG C
Under the conditions of 5 ~ 8h of stirring reaction；
Reaction finishes, and is separated by filtration, and is dried in vacuo after the washing of gained solids, obtains black solid thing；
d）Black solid thing after ultrasonic disperse, is fully reacted with N, the bromo-succinimides of N '-two at room temperature in ethanol；
Reaction finishes, and is separated by filtration, and is dried in vacuo after the washing of gained solids, obtained light/dark balance solids is load type platinum
Complex oxidant.
6. being easily recycled the preparation method of the load type platinum complex oxidant of recycling according to claim 5, it is special
Sign is step（a）In, the polystyrene microsphere is polyethyleneglycol modified polystyrene microsphere.
7. being easily recycled the preparation method of the load type platinum complex oxidant of recycling according to claim 5, it is special
Sign is step（a）In, the particle diameter of the polystyrene microsphere is 75 μm ~ 320 μm.
8. being easily recycled the preparation method of the load type platinum complex oxidant of recycling according to claim 5, it is special
Sign is step（c）In, the divalence platinum complex is cis Pt (en) Cl2。
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109836470A (en) * | 2019-03-22 | 2019-06-04 | 河北大学 | A method of being automatically synthesized disulfide bond in peptide molecule |
| CN109912477A (en) * | 2019-03-07 | 2019-06-21 | 河北大学 | A kind of selenomethionine oxide and preparation method thereof and the application in synthesis polypeptide intramolecular disulfide bond |
| US20230050515A1 (en) * | 2021-05-04 | 2023-02-16 | Cend Therapeutics, Inc. | iRGD-ANALOGS AND RELATED THERAPEUTIC METHODS |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000069889A1 (en) * | 1999-05-14 | 2000-11-23 | Merck Frosst Canada & Co. | Phosphonic and carboxylic acid derivatives as inhibitors of protein tyrosine phosphatase-1b (ptp-1b) |
| CN101490052A (en) * | 2006-05-12 | 2009-07-22 | 美瑞德生物工程公司 | Therapeutic compounds and their use in cancer |
| CN102596204A (en) * | 2009-07-06 | 2012-07-18 | 昂托瑞公司 | Novel nucleic acid prodrugs and methods use thereof |
| CN102812037A (en) * | 2009-10-30 | 2012-12-05 | 特兰齐姆制药公司 | Macrocyclic Ghrelin Receptor Antagonists And Inverse Agonists And Methods Of Using The Same |
| 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 |
| CN106565884A (en) * | 2016-10-19 | 2017-04-19 | 黑龙江大学 | Preparing method for polystyrene microsphere loaded nano-copper composite material |
| CN106866744A (en) * | 2017-02-20 | 2017-06-20 | 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 (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000069889A1 (en) * | 1999-05-14 | 2000-11-23 | Merck Frosst Canada & Co. | Phosphonic and carboxylic acid derivatives as inhibitors of protein tyrosine phosphatase-1b (ptp-1b) |
| CN101490052A (en) * | 2006-05-12 | 2009-07-22 | 美瑞德生物工程公司 | Therapeutic compounds and their use in cancer |
| CN102596204A (en) * | 2009-07-06 | 2012-07-18 | 昂托瑞公司 | Novel nucleic acid prodrugs and methods use thereof |
| CN102812037A (en) * | 2009-10-30 | 2012-12-05 | 特兰齐姆制药公司 | Macrocyclic Ghrelin Receptor Antagonists And Inverse Agonists And Methods Of Using The Same |
| 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 |
| CN106565884A (en) * | 2016-10-19 | 2017-04-19 | 黑龙江大学 | Preparing method for polystyrene microsphere loaded nano-copper composite material |
| CN106866744A (en) * | 2017-02-20 | 2017-06-20 | 河北大学 | A kind of tetravalence platinum complex oxidant and its preparation method and application |
Non-Patent Citations (11)
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109912477A (en) * | 2019-03-07 | 2019-06-21 | 河北大学 | A kind of selenomethionine oxide and preparation method thereof and the application in synthesis polypeptide intramolecular disulfide bond |
| 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 |
| CN109836470A (en) * | 2019-03-22 | 2019-06-04 | 河北大学 | A method of being automatically synthesized disulfide bond in peptide molecule |
| CN109836470B (en) * | 2019-03-22 | 2022-03-22 | 河北大学 | Method for automatically synthesizing polypeptide intramolecular disulfide bond |
| US20230050515A1 (en) * | 2021-05-04 | 2023-02-16 | Cend Therapeutics, Inc. | iRGD-ANALOGS AND RELATED THERAPEUTIC METHODS |
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