CN114315959A - Sipunculus nudus tripeptide, Sipunculus nudus tripeptide metal chelate and application thereof - Google Patents
Sipunculus nudus tripeptide, Sipunculus nudus tripeptide metal chelate and application thereof Download PDFInfo
- Publication number
- CN114315959A CN114315959A CN202111082550.9A CN202111082550A CN114315959A CN 114315959 A CN114315959 A CN 114315959A CN 202111082550 A CN202111082550 A CN 202111082550A CN 114315959 A CN114315959 A CN 114315959A
- Authority
- CN
- China
- Prior art keywords
- tripeptide
- sipunculus nudus
- sipunculus
- nudus
- ethyl alcohol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to the technical field of biological medicines, and particularly discloses sipunculus nudus tripeptide, a sipunculus nudus tripeptide metal chelate and application thereof. The amino acid sequence of the sipunculus nudus tripeptide is Ser-Arg-Pro. The sipunculus nudus tripeptide metal chelate is prepared by taking sipunculus nudus tripeptide as a raw material and reacting the raw material with zinc salt or magnesium salt. Researches show that the sipunculus nudus tripeptide and the sipunculus nudus tripeptide metal chelate not only have the antioxidant effect, but also have the hair regeneration promoting effect.
Description
Technical Field
The invention relates to the technical field of biomedicine, and particularly relates to sipunculus nudus tripeptide, a sipunculus nudus tripeptide metal chelate and application thereof.
Background
Along with the improvement of living standard and the increase of working pressure, the free radicals of the human body are generated more and more, and the threat to the health of human beings is more and more serious.
Free Radical (ROS) is a substance lacking electrons, and when the free radical in a human body is excessive, the free radical can rob the electrons of original healthy cells, so that the healthy cells are unbalanced and then a destructive chain reaction is generated, thereby accelerating aging, showing on the skin, the skin aging, and wrinkles and spots; manifested on the head, causing seborrheic dermatitis, causing alopecia; the heart and blood vessels show pathological states such as coronary heart disease and hypertension, hyperlipidemia and hyperglycemia, and cause various damages to the healthy body. While humans are unconsciously injured by free radicals every day, examples are: ultraviolet radiation in sunlight, computer radiation, fatigue, diabetes, cardiovascular system diseases, hypertension, hyperglycemia, hyperlipidemia, etc. Scavenging free radicals is therefore a task in human graduate.
The process from generation to decay of free radicals is the process of electron transfer, and free radicals damaging human health are almost related to oxygen-containing substances with stronger activity, so that the damage of the free radicals to the human body is actually an oxidation process, and the oxidation resistance is needed to reduce the damage of the free radicals.
Sandworm is a special marine product in Zhanjiang and is known as Sipunculus nudus. Sipunculus nudus can be used as medicine. Medicine property: sweet, salty and cold in nature. Enter spleen and stomach meridians. The functions and indications are as follows: to nourish yin and reduce pathogenic fire. Can be used for treating night sweat due to yin deficiency, hectic fever, cough due to pulmonary tuberculosis, and gingival swelling and pain. China medicinal marine life: nourishing yin to reduce pathogenic fire. Can be used for treating hectic fever due to yin deficiency, night sweat, pulmonary tuberculosis, cough, chest distress, and excessive phlegm. "Chinese animal medical record": to invigorate the spleen. It is used for treating nocturia. The Sipunculus nudus is rich in protein, polypeptide components and 17 amino acids, wherein the amino acid content necessary for human body is very high, and in addition, the Sipunculus nudus also contains 12 trace elements such as calcium, phosphorus, iron, zinc, manganese, magnesium and the like, and nutrient components such as cordycepin and the like with the effects of resisting oxidation, bacteria, radiation, virus, fatigue, cancer, immunity regulation and aging delay.
Therefore, the development of a polypeptide with an antioxidant effect by taking the sipunculus griseus as a research object has important application value.
Disclosure of Invention
In order to overcome at least one of the technical problems of the prior art, the present invention firstly provides a sipunculus nudus tripeptide. The sipunculus nudus tripeptide can be obtained by carrying out enzymolysis on sipunculus nudus; can also be synthesized by an artificial synthesis method. Research shows that the sipunculus nudus tripeptide has an anti-oxidation effect; particularly, the further research of the invention shows that the sipunculus nudus tripeptide also has the effect of promoting hair regeneration.
The technical problem to be solved by the invention is realized by the following technical scheme:
in a first aspect of the invention, the sipunculus nudus tripeptide is provided, and the amino acid sequence of the sipunculus nudus tripeptide is Ser-Arg-Pro (shown as SEQ ID No. 1).
The sipunculus nudus tripeptide can be obtained by carrying out enzymolysis on sipunculus nudus and can also be obtained by synthesizing by a conventional polypeptide synthesis method.
In a second aspect of the present invention, there is provided a sipunculus nudus tripeptide metal chelate prepared by a method comprising the steps of:
s01, dissolving Sipunculus nudus tripeptide with water;
s02, adding metal salt into water in which Sipunculus nudus tripeptide is dissolved, and carrying out chelation reaction in water bath;
s03, adding absolute ethyl alcohol into the reaction solution, stirring and centrifuging;
s04, placing the precipitate into absolute ethyl alcohol, and washing the precipitate in a shaking way;
s05, adding absolute ethyl alcohol into the washed precipitate, and freezing and storing at-15 to-25 ℃; after freezing and preservation, separating and precipitating to obtain the sipunculus nudus tripeptide metal chelate;
the metal salt in the step S02 is a zinc salt or a magnesium salt; if zinc salt is added, preparing Sipunculus nudus tripeptide zinc chelate; and if magnesium salt is added, preparing the sipunculus nudus tripeptide magnesium chelate.
The inventor further researches and surprisingly discovers that the Sipunculus nudus tripeptide zinc chelate and the Sipunculus nudus tripeptide magnesium chelate prepared by the method have far better antioxidant effect and hair regeneration promoting effect than the Sipunculus nudus tripeptide.
Preferably, the sipunculus nudus tripeptide zinc chelate is prepared by the following method:
s11, adding water into sipunculus nudus tripeptide, and adjusting the pH value to 5-6; stirring and dissolving;
s12, adding zinc salt into water in which Sipunculus nudus tripeptide is dissolved, and carrying out chelation reaction in a water bath at 50-60 ℃ for 30-50 min;
s13, adding absolute ethyl alcohol into the reaction solution, stirring and centrifuging; wherein the volume ratio of the absolute ethyl alcohol to the reaction liquid is 2-5: 1;
s14, placing the precipitate into absolute ethyl alcohol, and washing for 1-3 min in a shaking way;
s15, adding absolute ethyl alcohol into the washed precipitate, and freezing and storing at-15 to-25 ℃; after freezing and preservation, separating and precipitating to obtain the sipunculus nudus tripeptide metal chelate; wherein the dosage ratio of the washed precipitate to the absolute ethyl alcohol is 1g: 2-5 mL.
Further preferably, the mass ratio of the sipunculus nudus tripeptide to the water in the step S11 is 1: 20-30.
Most preferably, the mass ratio of sipunculus nudus tripeptide to water in step S11 is 1: 25.
Further preferably, the mass ratio of the sipunculus nudus tripeptide to the zinc salt in the step S12 is 2-3: 1.
Most preferably, the mass ratio of the sipunculus nudus tripeptide to the zinc salt in step S12 is 2.5: 1.
Preferably, the sipunculus nudus tripeptide magnesium chelate is prepared by the following method:
s21, adding water into sipunculus nudus tripeptide, and adjusting the pH value to 7; stirring and dissolving;
s22, adding magnesium salt into water in which Sipunculus nudus tripeptide is dissolved, and carrying out chelation reaction in water bath at 65-75 ℃ for 40-60 min;
s23, adding absolute ethyl alcohol into the reaction solution, stirring and centrifuging; wherein the volume ratio of the absolute ethyl alcohol to the reaction liquid is 2-5: 1;
s24, placing the precipitate into absolute ethyl alcohol, and washing for 1-3 min in a shaking way;
s25, adding absolute ethyl alcohol into the washed precipitate, and freezing and storing at the temperature of between 15 ℃ below zero and 25 ℃ below zero; after freezing and preservation, separating and precipitating to obtain the sipunculus nudus tripeptide metal chelate; wherein the dosage ratio of the washed precipitate to the absolute ethyl alcohol is 1g: 2-5 mL.
Further preferably, the mass ratio of the sipunculus nudus tripeptide to the water in the step S21 is 1: 25-35.
Most preferably, the mass ratio of the sipunculus nudus tripeptide to the water in the step S21 is 1: 25;
further preferably, the mass ratio of the sipunculus nudus tripeptide to the zinc salt in the step S22 is 2.5-3.5: 1.
Most preferably, the mass ratio of the sipunculus nudus tripeptide to the zinc salt in step S02 is 3: 1.
In a third aspect of the invention, there is provided a composition comprising a sipunculus nudus tripeptide and a sipunculus nudus tripeptide metal chelate.
Preferably, the mass ratio of the sipunculus nudus tripeptide to the sipunculus nudus tripeptide metal chelate is 1: 3-5.
During further research, the inventor surprisingly found that the combination of sipunculus nudus tripeptide and sipunculus nudus tripeptide metal chelate can generate synergistic antioxidation; so that the antioxidant effect of the composition is further better than that of the sipunculus nudus tripeptide or the sipunculus nudus tripeptide metal chelate.
In a fourth aspect of the invention, there is provided the use of the sipunculid tripeptide, the sipunculid tripeptide metal chelate or the composition as described above in the preparation of a product having a hair regrowth promoting effect or an antioxidant effect.
Preferably, the products include daily chemical products, medicines and health products.
Has the advantages that: the invention provides sipunculus nudus tripeptide with a brand new structure; the sipunculus nudus tripeptide not only has an antioxidant effect, but also has a hair regeneration promoting effect. Further research by the inventor shows that the Sipunculus nudus tripeptide zinc chelate and the Sipunculus nudus tripeptide magnesium chelate prepared by the method have far better antioxidant effect and hair regeneration promoting effect than Sipunculus nudus tripeptide. Therefore, the Sipunculus nudus tripeptide and/or the Sipunculus nudus tripeptide metal chelate can be used for preparing daily chemical products, medicines and health products, so that the daily chemical products, the medicines and the health products have the effects of resisting oxidation and promoting hair regeneration.
Drawings
FIG. 1 is a mass spectrum of Sipunculus nudus tripeptide.
FIG. 2 is a liquid chromatogram of Sipunculus nudus tripeptide.
Detailed Description
The present invention is further explained below with reference to specific examples, which are not intended to limit the present invention in any way.
Example 1 preparation of Sipunculus nudus tripeptide
(1) Pouring 250mg of fmoc-Ser (tbu) -OH resin into a reaction column, adding DMF, soaking for 30min, and pumping to dry; then adding piperidine DMF solution, oscillating and filtering out the solvent;
(2) adding a proper amount of deprotection solution (piperidine is 20% hexahydropyridine and 80% DMF) into a reaction column, introducing nitrogen, stirring for 30 minutes, pumping to dry, adding a proper amount of DMF into the reaction column, stirring for 2 minutes by nitrogen, pumping to dry, and repeating the operation for 6 times;
(3) adding Fmoc-Arg (pbf) -OH, a condensing agent HBTU and organic base NMM, stirring and agitating nitrogen for 30 minutes to carry out condensation reaction; wherein the molar dosage ratio of Fmoc-Arg (pbf) -OH, condensing agent HBTU and organic base NMM to Fmoc-Ser (tbu) -OH resin is 3:2.85:6: 1;
(4) after the condensation reaction is finished, pumping the solution in the reaction column, adding a proper amount of DMF for washing, pumping for 2 minutes by nitrogen, pumping, and repeating the operation for 3 times;
(5) adding Fmoc-Pro-OH for condensation reaction according to the steps 2-4; removing the protecting group Fmoc on the Fmoc-Pro-OH after the reaction is finished, and exposing the N end;
(6) and (3) pumping, adding a proper amount of methanol into the reaction column, pumping for 2 minutes by nitrogen, pumping, adding a proper amount of DCM, pumping for 2 minutes by nitrogen, and pumping for repeating the operation for 3 times. Finally, adding a proper amount of methanol into the reaction kettle, blowing with nitrogen for 2 minutes, pumping, repeating the operation for 2 times, putting the resin into a proper vessel, and placing the vessel into a vacuum drier for vacuum drying for 12 hours to be cut;
(7) the dried resin was charged into a suitable round-bottomed flask and the appropriate amount of cutting fluid (1g/10ml, composition 87.5% TFA + 5% thioanisole + 2.5% phenol + 2.5% EDT + 2.5% H) was added2O), placing the mixture in a constant temperature shaking table to oscillate for 2 hours at a constant temperature of 25 ℃; and (3) filtering: filtering resin particles by using a 50mL sand core funnel, pouring the filtrate into a glass container of a centrifuge tube, adding 6-8 times of volume of anhydrous ether, and stirring while adding, wherein the separated white solid is the required polypeptide crude product;
(8) subpackaging the white solid into a centrifuge tube, sealing the centrifuge tube, putting the centrifuge tube into a centrifuge, centrifuging for 3 minutes at the rotating speed of 4000r/min, taking out, pouring out supernatant liquid, adding diethyl ether, stirring uniformly by using a glass rod, and centrifuging again; the operation is repeated for 5 times;
(9) putting the polypeptide washed for 5 times into a vacuum drier for vacuum drying for 24 hours; further purifying with high performance liquid chromatography to obtain pure product of Sipunculus nudus tripeptide with amino acid sequence of Ser-Arg-Pro (see figure 2 for liquid chromatogram of Sipunculus nudus tripeptide with amino acid sequence of Ser-Arg-Pro);
the high performance liquid chromatography conditions are as follows:
the type of the chromatographic column: 250 x 4.6mm, Boston Green ODS-AQ;
the detection wavelength is as follows: 220 nm;
the flow rate is: 1.0 mL/min;
mobile phase a water containing 0.1% trifluoroacetic acid (TFA);
mobile phase B acetonitrile containing 0.1% trifluoroacetic acid (TFA);
gradient elution conditions: at 0.1min, the volume of mobile phase B is 5%; at 25min, the volume of mobile phase B increased to 30%; at 25.01min, the volume of mobile phase B increased to 100%; at 30min, the volume of mobile phase B was 100%.
The mass spectrum of Sipunculus nudus tripeptide with the amino acid sequence of Ser-Arg-Pro (see FIG. 1) shows that 359.3 of [ M + H]+An ion peak; the molecular weight of the tripeptide is 358, which is consistent with the molecular weight of the tripeptide Ser-Arg-Pro Sipunculus nudus, and the tripeptide Ser-Arg-Pro Sipunculus nudus is successfully synthesized.
Example 2 preparation of Sipunculus nudus tripeptide Zinc chelate
(1) Dissolving: putting the sipunculus nudus tripeptide prepared by the method in the example 1 into a beaker, adding ultrapure water (the weight ratio of the sipunculus nudus tripeptide to the ultrapure water is 1: 25), stirring, dissolving, and adjusting the pH value to 5.5;
(2) carrying out chelation reaction: placing the beaker in a water bath at 55 ℃, adding zinc sulfate (the mass ratio of Sipunculus nudus tripeptide: zinc is 2.5: 1) under stirring, and carrying out chelation reaction for 35 min. Standing at room temperature;
(3) washing: adding anhydrous ethanol into the reaction solution, washing, stirring for 10min, and centrifuging (5000r/min) for 10 min; wherein the volume ratio of the absolute ethyl alcohol to the reaction liquid is 2: 1;
(4) and (3) purification: adding absolute ethyl alcohol into the precipitate, washing for 2min with shaking, centrifuging (5000r/min) for 10min, repeating the above steps for 2 times to remove free Zn + +, adding absolute ethyl alcohol again, and placing in a refrigerator at-20 deg.C for 36 h; wherein the dosage ratio of the washed precipitate to the absolute ethyl alcohol is 1g:3 mL;
(5) and (3) freeze drying: and (4) placing the mixture after centrifugation in a freeze dryer, and freeze-drying to obtain white crystals, namely the sipunculus nudus tripeptide zinc chelate.
Example 3 preparation of Sipunculus nudus tripeptide magnesium chelate
(1) Dissolving: putting the sipunculus nudus tripeptide prepared by the method in the example 1 into a beaker, adding ultrapure water (the weight ratio of the sipunculus nudus tripeptide to the ultrapure water is 1: 30), stirring, dissolving, and adjusting the pH value to 7.0;
(2) carrying out chelation reaction: placing the beaker in 70 deg.C water bath, keeping the temperature for 10min, adding anhydrous MgCI under stirring2(the mass ratio of Sipunculus nudus tripeptide magnesium is 3: 1), carrying out chelation reaction for 45min, and standing at room temperature;
(3) washing: adding anhydrous ethanol into the reaction solution, stirring for 10min, and centrifuging (5000r/min) for 10 min; wherein the volume ratio of the absolute ethyl alcohol to the reaction liquid is 2: 1;
(4) and (3) purification: adding absolute ethyl alcohol into the precipitate, washing by shaking, centrifuging, separating supernatant, and detecting free Mg++If a white precipitate is formed, the above steps are repeated until no Mg is detected++Until the end; adding anhydrous ethanol again, and placing in a refrigerator at-20 deg.C for 36 h; wherein the dosage ratio of the washed precipitate to the absolute ethyl alcohol is 1g:3 mL;
(5) and (3) freeze drying: and (3) placing the precipitate in a freeze-drying machine, and freeze-drying to obtain white crystals, namely the sipunculus nudus tripeptide magnesium chelate.
Example 4 preparation of the composition
The sipunculus nudus tripeptide prepared in the example 1 and the sipunculus nudus tripeptide zinc chelate prepared in the example 2 are uniformly mixed according to the mass ratio of 1:4 to obtain the composition.
Example 5 preparation of the composition
The sipunculus nudus tripeptide prepared in the example 1 and the sipunculus nudus tripeptide magnesium chelate prepared in the example 3 are uniformly mixed according to the mass ratio of 1:4 to obtain the composition.
Experimental example 1 Oxidation resistance test
And evaluating the antioxidant capacity of Sipunculus nudus tripeptide and Sipunculus nudus tripeptide zinc chelate by adopting a DPPH method. The specific method comprises the following steps:
preparing 100 mu mol/L DPPH ethanol solution by using ethanol with the volume fraction of 90%; preparing 1mg/mL Sipunculus nudus tripeptide, Sipunculus nudus tripeptide zinc chelate and a composition by using water to obtain a sample solution to be detected; uniformly mixing 0.5mL of sample to be detected with 2.5mL of DPPH ethanol solution, incubating at 37 ℃ in a dark place for 30min, and then measuring the absorbance of the reaction solution at 517 nm; each sample is tested in parallel for 3 times, and the average value is taken; the formula for DPPH radical clearance is: DPPH radical scavenging ratio (%) - (1- (A)x-Ay)/AZ) ×100；
In the formula, AxThe absorbance of the sample solution to be detected after incubation of 0.5mL and 2.5mL of DPPH ethanol solution; a. theyThe absorbance of the sample solution to be detected after incubation of 0.5mL and 2.5mL of ethanol solution with volume fraction of 90%; a. theZThe absorbance was measured after incubation of 0.5mL of water with 2.5mL of ethanol DPPH solution. The test results are shown in Table 1.
TABLE 1 DPPH radical scavenging test results
| Sample to be tested | DPPH radical clearance rate |
| Example 1 preparation of tripeptide Sipunculus nudus | 62％ |
| EXAMPLE 2 Sipunculus nudus tripeptide Zinc chelate prepared | 85％ |
| EXAMPLE 3 Sipunculus nudus tripeptide magnesium chelate prepared | 81％ |
| Example 4 the resulting composition | 99％ |
| Example 5 the resulting composition | 95％ |
As can be seen from the DPPH free radical clearance test results in Table 1, the Sipunculus nudus tripeptide provided by the invention has 62% of DPPH free radical clearance at the concentration of 1 mg/mL; this indicates that it has some resistance to oxidation.
Further, the DPPH free radical clearance rate of the Sipunculus nudus tripeptide zinc chelate and the Sipunculus nudus tripeptide magnesium chelate prepared in the examples 2 and 3 reaches 85 percent and 81 percent; compared with sipunculus nudus tripeptide, the method has great improvement; this shows that the Sipunculus nudus tripeptide zinc chelate and the Sipunculus nudus tripeptide magnesium chelate prepared by the method have better antioxidant capacity than Sipunculus nudus tripeptide.
Further, the compositions prepared in examples 4 and 5 achieved DPPH radical scavenging rates of 99% and 95%; far higher than Sipunculus nudus tripeptide, Sipunculus nudus tripeptide zinc chelate and Sipunculus nudus tripeptide magnesium chelate; the result shows that after the sipunculus nudus tripeptide is combined with the sipunculus nudus tripeptide zinc chelate or the sipunculus nudus tripeptide magnesium chelate, the synergistic antioxidation effect can be generated between the sipunculus nudus tripeptide zinc chelate or the sipunculus nudus tripeptide magnesium chelate, and the antioxidation capability of the sipunculus nudus tripeptide zinc chelate or the sipunculus nudus tripeptide magnesium chelate is further improved.
Experimental example 2 ability to promote Hair regeneration
The backs of 12 SD rats were dehaired with 10% NaS solution and randomly divided into 4 groups of 3 rats each; wherein unhairing rats of the experimental group 1 are smeared with the sipunculus nudus tripeptide aqueous solution prepared in the example 1; the unhairing rat of the experimental group 2 is smeared with the sipunculus nudus tripeptide zinc chelate aqueous solution prepared in the example 2; the unhairing rat of the experimental group 3 is smeared with the sipunculus nudus tripeptide magnesium chelate aqueous solution prepared in the example 3; example 4 equal amounts of water were applied daily; the concentration of tripeptide and Sipunculus nudus tripeptide zinc chelate or Sipunculus nudus tripeptide magnesium chelate aqueous solution is 1mg/L, and the tripeptide zinc chelate or the Sipunculus nudus tripeptide magnesium chelate aqueous solution is smeared for 3 times every day, and 1mL is smeared every time; after 7 days, hair growth was observed; and measuring the average length of the growing hair; the results are shown in Table 2.
TABLE 2 ability to promote hair regrowth test
As can be seen from the experiment in Table 2, the tripeptide provided by the invention and Sipunculus nudus, tripeptide zinc chelate or Sipunculus nudus tripeptide magnesium chelate have the effect of promoting hair regeneration; and the Sipunculus nudus tripeptide zinc chelate or Sipunculus nudus tripeptide magnesium chelate has far better hair regeneration promoting effect than the tripeptide and the Sipunculus nudus.
EXAMPLE 6 preparation of Hair restorer
The formula is as follows: sipunculus nudus tripeptide zinc chelate 0.7%, Sipunculus nudus tripeptide magnesium chelate 0.35%, carbomer U-200.025%, glycerol 1.0%, triethanolamine 0.05%, ethylparaben 0.2%, 95% ethanol 10%, and sterile water in balance.
The preparation method comprises the following steps: adding a small amount of sterile deionized water into carbomer U20 and glycerol, and soaking for 12-15h to fully swell to obtain carbomer U20 mixed solution; adding triethanolamine, adjusting pH to 5.5-6.5, stirring to obtain carbomer U20 gel solution; adding the metal chelating peptide aqueous solution and the ethylparaben ethanol solution, stirring and mixing uniformly, finally adding sterile deionized water to full amount, and mixing uniformly to obtain the hair growth liquid.
Sequence listing
<110> ocean medicine institute of Zhanjiang province of Guangdong
<120> Sipunculus nudus tripeptide, Sipunculus nudus tripeptide metal chelate and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 3
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 1
Ser Arg Pro
1
Claims (10)
1. The sipunculus nudus tripeptide is characterized in that the amino acid sequence of the sipunculus nudus tripeptide is Ser-Arg-Pro.
2. A Sipunculus nudus tripeptide metal chelate is characterized by being prepared by a method comprising the following steps:
s01, dissolving Sipunculus nudus tripeptide with water;
s02, adding metal salt into water in which Sipunculus nudus tripeptide is dissolved, and carrying out chelation reaction in water bath;
s03, adding absolute ethyl alcohol into the reaction solution, stirring and centrifuging;
s04, placing the precipitate into absolute ethyl alcohol, and washing the precipitate in a shaking way;
s05, adding absolute ethyl alcohol into the washed precipitate, and freezing and storing at-15 to-25 ℃; after freezing and preservation, separating and precipitating to obtain the sipunculus nudus tripeptide metal chelate;
the metal salt in the step S02 is a zinc salt or a magnesium salt; if zinc salt is added, preparing Sipunculus nudus tripeptide zinc chelate; and if magnesium salt is added, preparing the sipunculus nudus tripeptide magnesium chelate.
3. The sipunculus nudus tripeptide metal chelate according to claim 2, wherein the sipunculus nudus tripeptide zinc chelate is prepared by the following method:
s11, adding water into sipunculus nudus tripeptide, and adjusting the pH value to 5-6; stirring and dissolving;
s12, adding zinc salt into water in which Sipunculus nudus tripeptide is dissolved, and carrying out chelation reaction in a water bath at 50-60 ℃ for 30-50 min;
s13, adding absolute ethyl alcohol into the reaction solution, stirring and centrifuging; wherein the volume ratio of the absolute ethyl alcohol to the reaction liquid is 2-5: 1;
s14, placing the precipitate into absolute ethyl alcohol, and washing for 1-3 min in a shaking way;
s15, adding absolute ethyl alcohol into the washed precipitate, and freezing and storing at-15 to-25 ℃; after freezing and preservation, separating and precipitating to obtain the sipunculus nudus tripeptide metal chelate; wherein the dosage ratio of the washed precipitate to the absolute ethyl alcohol is 1g: 2-5 mL.
4. The sipunculus nudus tripeptide metal chelate according to claim 3, wherein the mass ratio of the sipunculus nudus tripeptide to the water in the step S11 is 1: 20-30; most preferably, the mass ratio of the sipunculus nudus tripeptide to the water in the step S11 is 1: 25;
in the step S12, the mass ratio of the sipunculus nudus tripeptide to the zinc salt is 2-3: 1; most preferably, the mass ratio of the sipunculus nudus tripeptide to the zinc salt in step S12 is 2.5: 1.
5. The sipunculus nudus tripeptide metal chelate according to claim 2, wherein the sipunculus nudus tripeptide magnesium chelate is prepared by the following method:
s21, adding water into sipunculus nudus tripeptide, and adjusting the pH value to 7; stirring and dissolving;
s22, adding magnesium salt into water in which Sipunculus nudus tripeptide is dissolved, and carrying out chelation reaction in water bath at 65-75 ℃ for 40-60 min;
s23, adding absolute ethyl alcohol into the reaction solution, stirring and centrifuging; wherein the volume ratio of the absolute ethyl alcohol to the reaction liquid is 2-5: 1;
s24, placing the precipitate into absolute ethyl alcohol, and washing for 1-3 min in a shaking way;
s25, adding absolute ethyl alcohol into the washed precipitate, and freezing and storing at the temperature of between 15 ℃ below zero and 25 ℃ below zero; after freezing and preservation, separating and precipitating to obtain the sipunculus nudus tripeptide metal chelate; wherein the dosage ratio of the washed precipitate to the absolute ethyl alcohol is 1g: 2-5 mL.
6. The sipunculus nudus tripeptide metal chelate according to claim 5, wherein the mass ratio of the sipunculus nudus tripeptide to the water in the step S21 is 1: 25-35; most preferably, the mass ratio of the sipunculus nudus tripeptide to the water in the step S21 is 1: 25;
in the step S22, the mass ratio of the sipunculus nudus tripeptide to the zinc salt is 2.5-3.5: 1; most preferably, the mass ratio of the sipunculus nudus tripeptide to the zinc salt in step S02 is 3: 1.
7. A composition comprising a sipunculus nudus tripeptide and a sipunculus nudus tripeptide metal chelate.
8. The composition as claimed in claim 7, wherein the mass ratio of Sipunculus nudus tripeptide to Sipunculus nudus tripeptide metal chelate is 1: 3-5.
9. Use of a sipunculus nudus tripeptide, a sipunculus nudus tripeptide metal chelate or a composition according to any one of claims 1 to 8 for the preparation of a product having hair regeneration promoting or antioxidant effect.
10. The use according to claim 9, wherein the products include daily chemicals, pharmaceuticals and nutraceuticals.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111082550.9A CN114315959B (en) | 2021-09-15 | 2021-09-15 | Sipunculus nudus tripeptide, sipunculus nudus tripeptide metal chelate and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111082550.9A CN114315959B (en) | 2021-09-15 | 2021-09-15 | Sipunculus nudus tripeptide, sipunculus nudus tripeptide metal chelate and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114315959A true CN114315959A (en) | 2022-04-12 |
| CN114315959B CN114315959B (en) | 2023-09-29 |
Family
ID=81044481
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111082550.9A Active CN114315959B (en) | 2021-09-15 | 2021-09-15 | Sipunculus nudus tripeptide, sipunculus nudus tripeptide metal chelate and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114315959B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116813694A (en) * | 2023-06-21 | 2023-09-29 | 广东医科大学 | Sipunculus nudus peptide and preparation method and application thereof |
| CN117582485A (en) * | 2023-11-15 | 2024-02-23 | 湛江中心人民医院 | Application of Sipunculus nudus tripeptide in preparation of osteoporosis drugs |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112661811A (en) * | 2021-01-21 | 2021-04-16 | 南方海洋科学与工程广东省实验室(湛江) | Antihypertensive peptide, long-acting antihypertensive peptide and preparation method thereof |
| CN112679580A (en) * | 2021-01-14 | 2021-04-20 | 南方海洋科学与工程广东省实验室(湛江) | Sipunculus nudus oligopeptide capable of promoting bone development and preparation method and application thereof |
-
2021
- 2021-09-15 CN CN202111082550.9A patent/CN114315959B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112679580A (en) * | 2021-01-14 | 2021-04-20 | 南方海洋科学与工程广东省实验室(湛江) | Sipunculus nudus oligopeptide capable of promoting bone development and preparation method and application thereof |
| CN112661811A (en) * | 2021-01-21 | 2021-04-16 | 南方海洋科学与工程广东省实验室(湛江) | Antihypertensive peptide, long-acting antihypertensive peptide and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| PAPASSARA SANGTANOO等: "Anti-inflammatory action of two novel peptides derived from peanut worms (Sipunculus nudus) in lipopolysaccharide-induced RAW264.7 macrophages", 《FOOD & FUNCTION》, vol. 11, no. 1, pages 552 - 560 * |
| 孙雪萍等: "沙虫蛋白酶解产物抗氧化与免疫调节活性的研究", 《现代食品科技》, vol. 33, no. 04, pages 67 - 72 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116813694A (en) * | 2023-06-21 | 2023-09-29 | 广东医科大学 | Sipunculus nudus peptide and preparation method and application thereof |
| CN116813694B (en) * | 2023-06-21 | 2024-03-29 | 广东医科大学 | Sipunculus nudus peptide and preparation method and application thereof |
| CN117582485A (en) * | 2023-11-15 | 2024-02-23 | 湛江中心人民医院 | Application of Sipunculus nudus tripeptide in preparation of osteoporosis drugs |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114315959B (en) | 2023-09-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114315959B (en) | Sipunculus nudus tripeptide, sipunculus nudus tripeptide metal chelate and application thereof | |
| JP5057383B2 (en) | Novel mucin-type glycoprotein and its use | |
| CN100427502C (en) | Antineoplastic oligopeptide and its preparation method and application | |
| CN110894225B (en) | Large-scale preparation and purification method and application of mu-conopeptide | |
| WO2010104147A1 (en) | Antioxidant agent | |
| CN107573408B (en) | Synthetic method of high-purity leuprorelin | |
| NZ509622A (en) | Peptides based on the sequence of human lactoferrin and their use | |
| CN110066321A (en) | A kind of anti-bacterial hydrogel and its preparation method and application | |
| AU708084B2 (en) | Peptide and a method of obtaining it | |
| CN1865283B (en) | Solid phase polypeptide synthesis preparation method for salcatonin | |
| CN111518164A (en) | ACE inhibitory peptide P2, application thereof and preparation method thereof | |
| CN108070030B (en) | Preparation method of loxenapeptide and analogue thereof | |
| CN109336953B (en) | Tartary buckwheat antioxidant peptide and preparation method and application thereof | |
| CN107759660A (en) | A kind of liquid-solid phase synthetic method of tripeptides 29 | |
| CN111499691B (en) | ACE inhibitory peptide P1, application thereof and preparation method thereof | |
| CN1199986C (en) | Trivalent iron porphrin and its derivative-short peptide compound and its synthesis | |
| CN114209636B (en) | Composition and application thereof in preparing daily chemical products with hair growth and/or hair blacking effects | |
| CN113880916B (en) | Yak skin antioxidant polypeptide and preparation method and application thereof | |
| CN107722108B (en) | Liquid phase synthesis method of acetyl tetrapeptide-9 | |
| CN110520162B (en) | Combination of salicylic acid and peptide | |
| CN110655553A (en) | ACE inhibitory peptide derived from sesame, preparation method and application thereof in preparation of antihypertensive drugs | |
| CN113940961B (en) | Traditional Chinese medicine extract, composition and application thereof in preparation of toothpaste or mouthwash with tooth cleaning and solidifying effects | |
| NEUGEBAUER et al. | Peptide synthesis on chitin | |
| CN117088941B (en) | Tripeptide derivative, composition and application thereof | |
| CN117045534B (en) | Novel use of hexapeptide |
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 |