A method for preparing a cosmetic peptide prolyl-prolyl-lysine, a condensing agent and applications thereof

Abstract

The application belongs to the technical field of chemical industry and discloses a preparation method, a condensing agent and application of a cosmetic peptide prolyl-prolyl-lysine.The method uses proline as a starting material to generate proline with a benzyl-protected amino group by combining with bromobenzene;then, in water, sodium salt is formed by using sodium bicarbonate as a condensing agent to form an active lipid, which is directly combined with unprotected proline to generate Bn-Pro-Pro-OH after reaction;then, Bn-Pro-Pro-OH is used in water, sodium salt is formed by using sodium bicarbonate, an active lipid is formed by using a condensing agent, and N6-Cbz-L-lysine is combined to generate Bn-Pro-Pro-Lys(Z)-OH;finally, hydrogenation is performed to generate the cosmetic peptide prolyl-prolyl-lysine.The method has the advantages that the product purity of the prolyl-prolyl-lysine tripeptide obtained by the method is high, the raw material cost is low, the operation is simple, the method is suitable for industrial production, and the method can meet the needs of the future skin care product industry.

Description

Technical Field

[0001] This invention belongs to the field of chemical technology, and in particular to a method for preparing the cosmetic peptide prolyl-prolyl-lysine, a condensing agent, and its application. Background Technology

[0002] Prolyl-prolyl-lysine (Pro-Pro-Lys), Cas: 85807-08-5, tripeptide-42 fragment is found in dairy products and casein. It has blood-activating properties, helps improve dark circles and skin tone, and enhances skin firmness, elasticity, and suppleness, making it suitable as an anti-wrinkle conditioning agent. It is a cosmetic raw material with significant application value. However, there are currently no literature reports on the preparation method of prolyl-prolyl-lysine, which causes inconvenience in its use. Summary of the Invention

[0003] The purpose of this invention is to overcome the shortcomings of the prior art and provide a method for preparing the cosmetic peptide prolyl-prolyl-lysine, a condensing agent, and its application.

[0004] The technical solution adopted by this invention to solve its technical problem is:

[0005] A condensing agent for preparing the cosmetic peptide prolyl-prolyl-lysine, the structural formula of which is shown below:

[0006] .

[0007] The application of the condensing agent described above in the preparation of peptide prolyl-prolyl-lysine.

[0008] The preparation method of the condensing agent as described above includes the following steps:

[0009] (1) Preparation of 4,6-dimethoxy-1,3,5-triazine-2-ylmethanesulfonate

[0010] Dichloromethane and 4,6-dimethoxy-1,3,5-triazine-2(1H)-one were added to a stirred reactor that had been fully replaced with N2. The temperature was lowered to 0°C, and methanesulfonyl chloride was slowly added dropwise. After stirring for 30 min, the temperature was raised to 38-40°C and the reaction was continued for 12 hours. The system was concentrated to 10% of its original volume and the temperature was controlled at 35-40°C to obtain 4,6-dimethoxy-1,3,5-triazine-2-ylmethanesulfonate.

[0011] The ratio of dichloromethane: 4,6-dimethoxy-1,3,5-triazine-2(1H)-one: methanesulfonyl chloride in mL:g:g is 50:5:3.6.

[0012] (2) Preparation of N,N-bis(4,6-dimethoxy-1,3,5-triazine-2-yl)-N,N,N,N-tetramethylethane-1,2-diammonium dimethanesulfonate

[0013] Dichloromethane and 4,6-dimethoxy-1,3,5-triazin-2-yl methanesulfonate were added to a stirred reactor that had been fully replaced with N2. The temperature was lowered to 0°C and controlled at 0-5°C. Tetramethylethylenediamine was added dropwise, and the mixture was stirred for 1 hour. After the reaction was completed, the system was concentrated to 20% of its original volume and the temperature was controlled at 35-40°C. Then, methyl tert-butyl ether was added and the mixture was stirred and filtered. The mixture was dried at 35-40°C to obtain N,N-bis(4,6-dimethoxy-1,3,5-triazin-2-yl)-N,N,N,N-tetramethylethane-1,2-diammonium dimethanesulfonate, which is the condensing agent.

[0014] The ratio of dichloromethane: 4,6-dimethoxy-1,3,5-triazine-2-ylmethanesulfonate: tetramethylethylenediamine: methyl tert-butyl ether in mL: g: g: ml is 50:5:1.2:50.

[0015] A method for preparing the cosmetic peptide prolyl-prolyl-lysine using the condensing agent described above is described. The method uses proline as a starting material, reacting it with benzyl bromide to form proline with a benzyl-protected amino group. Then, in water, sodium bicarbonate is used to form a sodium salt, and a condensing agent is used to form an active lipid, which directly reacts with unprotected proline. After the reaction is complete, Bn-Pro-Pro-OH is obtained. Then, Bn-Pro-Pro-OH is used in water, with sodium bicarbonate forming a sodium salt, and a condensing agent is used to form an active lipid, which is then condensed with N6-Cbz-L-lysine to obtain Bn-Pro-Pro-Lys(Z)-OH. Finally, hydrogenation is performed to obtain the cosmetic peptide prolyl-prolyl-lysine.

[0016] Furthermore, the structural formula of the cosmetic peptide prolyl-prolyl-lysine is shown below:

[0017] .

[0018] Furthermore, it includes the following steps:

[0019] (1) Preparation of Bn-Pro-OH

[0020] Water and proline were added to a stirred reactor that had been fully replaced with N2. The temperature was lowered to 10°C, sodium carbonate was added, and the reaction was stirred for 30 min. Then benzyl bromide was added dropwise, and the reaction was stirred for 12 min until the reaction was complete. The pH of the system was adjusted to 6.5-7.0 with 6N hydrochloric acid, filtered, and dried at 40-45°C to obtain Bn-Pro-OH.

[0021] (2) Preparation of Bn-Pro-Pro-OH

[0022] Add water and Bn-Pro-OH to a stirred reactor, slowly add sodium bicarbonate, control the temperature to 0-5℃ and add condensing agent, stir and react for 1 hour, add proline, stir and react for 2 hours. After the reaction is complete, adjust the pH to 6.5-7.0 with 6N hydrochloric acid, filter, and dry at 30-35℃ to obtain Bn-Pro-Pro-OH.

[0023] (3) Preparation of Bn-Pro-Pro-Lys(Z)-OH

[0024] Water and Bn-Pro-Pro-OH were added to a stirred reactor, sodium bicarbonate was slowly added, the temperature was controlled at 0-5℃, a condensing agent was added, and the reaction was stirred for 1 hour. H-Lys(Z)-OH was added, and the reaction was stirred for 2 hours. After the reaction was completed, the pH was adjusted to 6.5-7.0 with 6N hydrochloric acid, filtered, and dried at 30-35℃ to obtain Bn-Pro-Pro-Lys(Z)-OH.

[0025] (4) Preparation of H-Pro-Pro-Lys-OH

[0026] Purified water and Bn-Pro-Pro-Lys(Z)-OH were added to a stirred reactor that had been fully purged with N2. The temperature was controlled at 20-25℃, and Pd / C with a palladium content of 10% was added. The reactor was then purged with hydrogen three times and reacted under hydrogen protection. The temperature was controlled at 20-25℃ for 24 hours. After filtration, the system was concentrated to 20% of its original volume, and the temperature was controlled at 35-40℃. Ethanol was then added and the mixture was stirred. The mixture was filtered and dried at 30-35℃ to obtain H-Pro-Pro-Lys-OH, which yielded the beauty peptide prolyl-prolyl-lysine.

[0027] Furthermore, in step (1), the ratio of water:proline:sodium carbonate:benzyl bromide in mL:g:g:g is 50:5:4.6:7.4;

[0028] In step (2), the ratio of water:Bn-Pro-OH:sodium bicarbonate:condensing agent:proline in mL:g:g:g:g is 50:3.5:2.8:5:1.9.

[0029] In step (3), the ratio of water:Bn-Pro-Pro-OH:sodium bicarbonate:condensing agent:H-Lys(Z)-OH in mL:g:g:g:g is 50:3:1.7:2.9:2.7.

[0030] In step (4), the ratio of purified water:Bn-Pro-Pro-Lys(Z)-OH:Pd / C:ethanol (ml:g:g:ml) is 50:2:0.2:100.

[0031] The preparation method described above is applied in the preparation of the cosmetic peptide prolyl-prolyl-lysine.

[0032] The application of the beauty peptide prolyl-prolyl-lysine prepared by the method described above in the preparation of skin care products.

[0033] The advantages and positive effects of this invention are as follows:

[0034] 1. This invention uses readily available and inexpensive proline as a starting material. It reacts with benzyl bromo to form proline with a benzyl-protected amino group, enhancing its solubility in water after sodium salt formation. Then, in water, sodium bicarbonate is used to form a sodium salt, and a novel condensing agent is used to form an active lipid. This active lipid reacts directly with unprotected proline, resulting in a good reaction, easy purification, and avoiding the use of organic solvents. After the reaction, Bn-Pro-Pro-OH is obtained. Bn-Pro-Pro-OH is then reacted in water with sodium bicarbonate to form a sodium salt, and a self-made novel condensing agent is used to form an active lipid. This active lipid is then condensed with N6-Cbz-L-lysine to obtain Bn-Pro-Pro-Lys(Z)-OH, showing a good reaction and avoiding the use of organic solvents. Finally, hydrogenation yields a prolyl-prolyl-lysine tripeptide. The product has high purity, low raw material cost, and simple operation, making it suitable for industrial production and meeting the future needs of the skincare industry.

[0035] 2. The method of this invention yields a novel condensing agent that is inexpensive, readily available, safe and easy to operate, and highly tolerant of reaction conditions. Furthermore, post-reaction processing should be simple, allowing for the removal of byproducts through simple extraction without producing any toxic residues, especially in the later stages of the production process.

[0036] 3. The unique feature of this invention lies in the use of a self-made, highly efficient catalyst, whose selectivity for amine groups is far greater than that for hydroxyl groups. Furthermore, it exhibits excellent stability to both air and water, making it ideal for long-distance transportation and storage. It is also suitable for use with water as the reaction medium, generating less waste, and its experimental operation is significantly superior to existing published methods. Most existing condensing agents require the protection of the carboxylic acid of another amino acid, followed by deprotection after the reaction, making the reaction operation rather cumbersome. This condensing agent significantly reduces production costs and is suitable for industrial-scale scaling. Attached Figure Description

[0037] Figure 1 The NMR spectrum of the target product TM in this invention;

[0038] Figure 2 This is the mass spectrometry spectrum of the target product TM in this invention.

[0039] Figure 3 This is the liquid phase spectrum of the target product TM in this invention. Detailed Implementation

[0040] The present invention will be further described below with reference to the embodiments. The following embodiments are descriptive and not limiting, and should not be used to limit the scope of protection of the present invention.

[0041] The various experimental operations involved in the specific embodiments are all conventional techniques in the field. For parts not specifically annotated in this document, those skilled in the art can refer to various commonly used reference books, scientific and technological documents or related instructions and manuals prior to the filing date of this invention to carry out the operations.

[0042] A condensing agent for preparing the cosmetic peptide prolyl-prolyl-lysine, the structural formula of which is shown below:

[0043] .

[0044] The application of the condensing agent described above in the preparation of peptide prolyl-prolyl-lysine.

[0045] The preparation method of the condensing agent as described above includes the following steps:

[0046] (1) Preparation of 4,6-dimethoxy-1,3,5-triazine-2-ylmethanesulfonate

[0047] Dichloromethane and 4,6-dimethoxy-1,3,5-triazine-2(1H)-one were added to a stirred reactor that had been fully replaced with N2. The temperature was lowered to 0°C, and methanesulfonyl chloride was slowly added dropwise. After stirring for 30 min, the temperature was raised to 38-40°C and the reaction was continued for 12 hours. The system was concentrated to 10% of its original volume and the temperature was controlled at 35-40°C to obtain 4,6-dimethoxy-1,3,5-triazine-2-ylmethanesulfonate.

[0048] The ratio of dichloromethane: 4,6-dimethoxy-1,3,5-triazine-2(1H)-one: methanesulfonyl chloride in mL:g:g is 50:5:3.6.

[0049] (2) Preparation of N,N-bis(4,6-dimethoxy-1,3,5-triazine-2-yl)-N,N,N,N-tetramethylethane-1,2-diammonium dimethanesulfonate

[0050] Dichloromethane and 4,6-dimethoxy-1,3,5-triazin-2-yl methanesulfonate were added to a stirred reactor that had been fully replaced with N2. The temperature was lowered to 0°C and controlled at 0-5°C. Tetramethylethylenediamine was added dropwise, and the mixture was stirred for 1 hour. After the reaction was completed, the system was concentrated to 20% of its original volume and the temperature was controlled at 35-40°C. Then, methyl tert-butyl ether was added and the mixture was stirred and filtered. The mixture was dried at 35-40°C to obtain N,N-bis(4,6-dimethoxy-1,3,5-triazin-2-yl)-N,N,N,N-tetramethylethane-1,2-diammonium dimethanesulfonate, which is the condensing agent.

[0051] The ratio of dichloromethane: 4,6-dimethoxy-1,3,5-triazine-2-ylmethanesulfonate: tetramethylethylenediamine: methyl tert-butyl ether in mL: g: g: ml is 50:5:1.2:50.

[0052] A method for preparing the cosmetic peptide prolyl-prolyl-lysine using the condensing agent described above is described. The method uses proline as a starting material, reacting it with benzyl bromide to form proline with a benzyl-protected amino group. Then, in water, sodium bicarbonate is used to form a sodium salt, and a condensing agent is used to form an active lipid, which directly reacts with unprotected proline. After the reaction is complete, Bn-Pro-Pro-OH is obtained. Then, Bn-Pro-Pro-OH is used in water, with sodium bicarbonate forming a sodium salt, and a condensing agent is used to form an active lipid, which is then condensed with N6-Cbz-L-lysine to obtain Bn-Pro-Pro-Lys(Z)-OH. Finally, hydrogenation is performed to obtain the cosmetic peptide prolyl-prolyl-lysine.

[0053] Furthermore, the structural formula of the cosmetic peptide prolyl-prolyl-lysine is shown below:

[0054] .

[0055] Furthermore, it includes the following steps:

[0056] (1) Preparation of Bn-Pro-OH

[0057] Water and proline were added to a stirred reactor that had been fully replaced with N2. The temperature was lowered to 10°C, sodium carbonate was added, and the reaction was stirred for 30 min. Then benzyl bromide was added dropwise, and the reaction was stirred for 12 min until the reaction was complete. The pH of the system was adjusted to 6.5-7.0 with 6N hydrochloric acid, filtered, and dried at 40-45°C to obtain Bn-Pro-OH.

[0058] (2) Preparation of Bn-Pro-Pro-OH

[0059] Add water and Bn-Pro-OH to a stirred reactor, slowly add sodium bicarbonate, control the temperature to 0-5℃ and add condensing agent, stir and react for 1 hour, add proline, stir and react for 2 hours. After the reaction is complete, adjust the pH to 6.5-7.0 with 6N hydrochloric acid, filter, and dry at 30-35℃ to obtain Bn-Pro-Pro-OH.

[0060] (3) Preparation of Bn-Pro-Pro-Lys(Z)-OH

[0061] Water and Bn-Pro-Pro-OH were added to a stirred reactor, sodium bicarbonate was slowly added, the temperature was controlled at 0-5℃, a condensing agent was added, and the reaction was stirred for 1 hour. H-Lys(Z)-OH was added, and the reaction was stirred for 2 hours. After the reaction was completed, the pH was adjusted to 6.5-7.0 with 6N hydrochloric acid, filtered, and dried at 30-35℃ to obtain Bn-Pro-Pro-Lys(Z)-OH.

[0062] (4) Preparation of H-Pro-Pro-Lys-OH

[0063] Purified water and Bn-Pro-Pro-Lys(Z)-OH were added to a stirred reactor that had been fully purged with N2. The temperature was controlled at 20-25℃, and Pd / C with a palladium content of 10% was added. The reactor was then purged with hydrogen three times and reacted under hydrogen protection. The temperature was controlled at 20-25℃ for 24 hours. After filtration, the system was concentrated to 20% of its original volume, and the temperature was controlled at 35-40℃. Ethanol was then added and the mixture was stirred. The mixture was filtered and dried at 30-35℃ to obtain H-Pro-Pro-Lys-OH, which yielded the beauty peptide prolyl-prolyl-lysine.

[0064] Furthermore, in step (1), the ratio of water:proline:sodium carbonate:benzyl bromide in mL:g:g:g is 50:5:4.6:7.4;

[0065] In step (2), the ratio of water:Bn-Pro-OH:sodium bicarbonate:condensing agent:proline in mL:g:g:g:g is 50:3.5:2.8:5:1.9.

[0066] In step (3), the ratio of water:Bn-Pro-Pro-OH:sodium bicarbonate:condensing agent:H-Lys(Z)-OH in mL:g:g:g:g is 50:3:1.7:2.9:2.7.

[0067] In step (4), the ratio of purified water:Bn-Pro-Pro-Lys(Z)-OH:Pd / C:ethanol (ml:g:g:ml) is 50:2:0.2:100.

[0068] The preparation method described above is applied in the preparation of the cosmetic peptide prolyl-prolyl-lysine.

[0069] The application of the beauty peptide prolyl-prolyl-lysine prepared by the method described above in the preparation of skin care products.

[0070] Specifically, the relevant preparation and testing methods are as follows:

[0071] This invention uses a novel condensing agent to react in water to generate a protected tripeptide, and then removes the protection to obtain prolyl-prolyl-lysine.

[0072] The catalytic mechanism of this novel condensing agent is as follows: firstly, the free carboxyl group attacks the catalyst to form an active ester, which has high activity and very high selectivity for amino groups. Subsequently, the amino group attacks the active ester to generate the target compound.

[0073]

[0074] In this invention, 4,6-dimethoxy-1,3,5-triazine-2(1H)-one and methanesulfonyl chloride are used to form an active ester, which is then reacted with tetramethylethylenediamine to form an onium salt, thereby obtaining a condensing agent.

[0075] This invention involves reacting benzyl bromide with proline to generate benzyl-protected proline. The benzyl-protected proline (Bn-Pro-OH) is then reacted with water to form a sodium salt using sodium bicarbonate, followed by the formation of an active lipoprotein using a novel self-made condensing agent. This active lipoprotein is then reacted with proline. After the reaction is complete, Bn-Pro-Pro-OH is obtained. This Bn-Pro-Pro-OH is then reacted with water to form a sodium salt using sodium bicarbonate, followed by the formation of an active lipoprotein using a novel self-made condensing agent. This active lipoprotein is then condensed with N6-Cbz-L-lysine to obtain Bn-Pro-Pro-Lys(Z)-OH. Finally, hydrogenation in water yields a prolyl-prolyl-lysine tripeptide.

[0076] Example 1

[0077] Preparation of novel condensing agents:

[0078] The structure is as follows:

[0079]

[0080] The preparation method is as follows:

[0081] (1) Preparation of 4,6-dimethoxy-1,3,5-triazine-2-ylmethanesulfonate

[0082] The preparation method is as follows:

[0083] Dichloromethane (50 mL) and 4,6-dimethoxy-1,3,5-triazine-2(1H)-one (5 g, 31.8 mmol) were added to a stirred 100 mL reactor fully purged with N2. The mixture was cooled to 0 °C, and methanesulfonyl chloride (3.6 g, 31.8 mmol) was slowly added dropwise. After stirring for 30 min, the temperature was raised to 38-40 °C and the reaction was continued for 12 h. The system was concentrated to 10% of its original volume, and the temperature was controlled at 35-40 °C to give 7.47 g of 4,6-dimethoxy-1,3,5-triazine-2-ylmethanesulfonate, with a yield of 100%.

[0084] (2) Preparation of N,N-bis(4,6-dimethoxy-1,3,5-triazine-2-yl)-N,N,N,N-tetramethylethane-1,2-diammonium dimethanesulfonate

[0085] The preparation method is as follows:

[0086] In a stirred 100 mL reactor fully purged with N2, dichloromethane (50 mL) and 4,6-dimethoxy-1,3,5-triazin-2-yl methanesulfonate (5 g, 21.2 mmol) were added. The mixture was cooled to 0 °C and then controlled at 0–5 °C. Tetramethylethylenediamine (1.2 g, 10.6 mmol) was added dropwise. The reaction was stirred for 1 hour. After the reaction was complete, the system was concentrated to 20% of its original volume, and the temperature was controlled at 35–40 °C. Then, 50 mL of methyl tert-butyl ether (methyl tert-butyl ether) was added and the mixture was stirred. The mixture was filtered and dried at 35–40 °C to obtain 6.21 g of N,N-bis(4,6-dimethoxy-1,3,5-triazin-2-yl)-N,N,N,N-tetramethylethane-1,2-diammonium dimethanesulfonate, with a yield of 94%.

[0087] Example 2

[0088] Preparation of prolyl-prolyl-lysine tripeptide:

[0089] The structure is as follows:

[0090]

[0091] The preparation method is as follows:

[0092] (1) Preparation of Bn-Pro-OH

[0093] The preparation method is as follows:

[0094] Water (50 mL) and proline (5 g, 43.4 mmol) were added to a stirred 100 mL reactor fully purged with N2. The mixture was cooled to 10 °C, and sodium carbonate (4.6 g, 43.4 mmol) was added. After stirring for 30 min, benzyl bromide (7.4 g, 43.4 mmol) was added dropwise, and the mixture was stirred for 12 min until the reaction was complete. The pH of the system was adjusted to 6.5-7.0 with 6N hydrochloric acid, filtered, and dried at 40-45 °C to obtain 7.9 g of Bn-Pro-OH, with a yield of 89%.

[0095] (2) Preparation of Bn-Pro-Pro-OH

[0096] The preparation method is as follows:

[0097] Water (50 mL), Bn-Pro-OH (3.5 g, 17.1 mmol), and sodium bicarbonate (2.8 g, 34.1 mmol) were added slowly to a 500 mL stirred reactor. The temperature was controlled at 0–5 °C, and N,N-bis(4,6-dimethoxy-1,3,5-triazin-2-yl)-N,N,N,N-tetramethylethane-1,2-diammonium dimethanesulfonate (5 g, 8.6 mmol) was added. The reaction was stirred for 1 hour, then proline (1.9 g, 17.1 mmol) was added, and the reaction was stirred for another 2 hours. After the reaction was complete, the pH was adjusted to 6.5–7.0 with 6N hydrochloric acid. The mixture was filtered and dried at 30–35 °C to obtain 3.9 g of Bn-Pro-Pro-OH, with a yield of 76%.

[0098] (3) Preparation of Bn-Pro-Pro-Lys(Z)-OH

[0099] The preparation method is as follows:

[0100] Water (50 mL), Bn-Pro-Pro-OH (3 g, 9.9 mmol), and sodium bicarbonate (1.7 g, 19.8 mmol) were slowly added to a 500 mL stirred reactor. The temperature was controlled at 0-5 °C, and N,N-bis(4,6-dimethoxy-1,3,5-triazin-2-yl)-N,N,N,N-tetramethylethane-1,2-diammonium dimethanesulfonate (2.9 g, 5.0 mmol) was added. The reaction was stirred for 1 hour, and H-Lys(Z)-OH (2.7 g, 9.9 mmol) was added. The reaction was stirred for another 2 hours. After the reaction was complete, the pH was adjusted to 6.5-7.0 with 6N hydrochloric acid. The mixture was filtered and dried at 30-35 °C to obtain 3.9 g of Bn-Pro-Pro-Lys(Z)-OH, with a yield of 70%.

[0101] (4) Preparation of H-Pro-Pro-Lys-OH

[0102] The preparation method is as follows:

[0103] In a stirred 500 mL reactor fully purified with N2, 50 mL of purified water and 2 g (3.5 mmol) of Bn-Pro-Pro-Lys(Z)-OH were added. The temperature was controlled at 20-25 °C, and 0.2 g of Pd / C with a palladium content of 10% was added. The mixture was then purged three times with hydrogen under hydrogen protection. The reaction was carried out at 20-25 °C for 24 hours. After filtration, the system was concentrated to 20% of its original volume, and the temperature was controlled at 30-35 °C. Then, 100 mL of ethanol was added, the mixture was stirred, filtered, and dried to obtain 1.6 g of H-Pro-Pro-Lys-OH, with a yield of 95%. This yielded the prolyl-prolyl-lysine tripeptide.

[0104] The liquid phase spectrum of the target product is as follows: Figure 3 As shown, the liquid phase purity reached 99.33%, which is high and meets the requirements of the skincare industry. The 1H NMR spectrum of the target product is shown below. Figure 1 As shown, the NMR spectrum is clean with no impurity peaks, and the characteristic peaks have the correct structure. The mass spectrum of the target product is shown below. Figure 2 As shown, the characteristic peak structure is correct.

[0105] This invention utilizes readily available and inexpensive 4,6-dimethoxy-1,3,5-triazine-2(1H)-one to prepare a novel self-made condensing agent through a simple two-step reaction. Then, using Bn-Pro and proline as starting materials, the sodium salt of Bn-Pro is soluble in water and undergoes a condensation reaction with proline in water, yielding Bn-Pro-Pro. This sodium salt of Bn-Pro-Pro is then soluble in water and undergoes a condensation reaction with N6-Cbz-L-lysine in water, yielding Bn-Pro-Pro-Lys(Z)-OH. Compared to conventional acid-amine condensation reactions, this condensation reaction does not require the addition of a solvent, and the carboxylic acid does not require protection, reducing material costs and waste generation. Finally, hydrogenation yields a prolyl-prolyl-lysine tripeptide product with high purity, meeting the needs of the skincare industry. The overall yield of the four steps is 44%, the route is short, the operation is simple, and waste is minimal, making it suitable for large-scale industrial production.

[0106] The specific methods of liquid chromatography are as follows:

[0107] 1. Instruments:

[0108] Analytical instruments: Agilent 1260 liquid chromatograph; analytical balance: AR224CN

[0109] 2. Chromatographic column: Nouryon Kromasil C18 5.0um 4.6 150mm

[0110] 3. Reagents:

[0111] Sodium heptanesulfonate: ion-pairing reagent, ≥99.5%

[0112] Trifluoroacetic acid: chromatographic grade, ≥99%

[0113] Acetonitrile: chromatographic grade, ≥99%

[0114] Pure water: purified water or distilled water

[0115] 4. Chromatographic conditions:

[0116]

[0117] 5. Preparation of the mobile phase:

[0118] Mobile phase A: 10 mmol / L sodium heptanesulfonate aqueous solution with 0.1% TFA

[0119] Weigh approximately 2.2g of sodium heptanesulfonate into a mobile phase bottle containing 1L of water, add 1ml of trifluoroacetic acid, and then sonicate and filter.

[0120] Mobile phase B: Acetonitrile

[0121] Measure an appropriate amount of acetonitrile into a suitable mobile phase bottle.

[0122] 6. Solution preparation:

[0123] Preparation of test solution (2.0 mg / ml)

[0124] Weigh approximately 20 mg of the test sample into a 10 ml volumetric flask, dilute with diluent, and make up to volume and mix well.

[0125] Although embodiments of the invention have been disclosed for illustrative purposes, those skilled in the art will understand that various substitutions, variations, and modifications are possible without departing from the spirit and scope of the invention and the appended claims. Therefore, the scope of the invention is not limited to the contents disclosed in the embodiments.

Claims

1. A condensing agent for preparing the cosmetic peptide prolyl-prolyl-lysine, characterized in that: The structural formula of the condensing agent is shown below: 。 2. The application of the condensing agent as described in claim 1 in the preparation of peptide prolyl-prolyl-lysine.

3. The method for preparing the condensing agent as described in claim 1, characterized in that: Includes the following steps: (1) Preparation of 4,6-dimethoxy-1,3,5-triazine-2-ylmethanesulfonate Dichloromethane and 4,6-dimethoxy-1,3,5-triazine-2(1H)-one were added to a stirred reactor that had been fully replaced with N2. The temperature was lowered to 0°C, and methanesulfonyl chloride was slowly added dropwise. After stirring for 30 min, the temperature was raised to 38-40°C and the reaction was continued for 12 hours. The system was concentrated to 10% of its original volume and the temperature was controlled at 35-40°C to obtain 4,6-dimethoxy-1,3,5-triazine-2-ylmethanesulfonate. The ratio of dichloromethane: 4,6-dimethoxy-1,3,5-triazine-2(1H)-one: methanesulfonyl chloride in mL:g:g is 50:5:3.

6. (2) Preparation of N,N-bis(4,6-dimethoxy-1,3,5-triazine-2-yl)-N,N,N,N-tetramethylethane-1,2-diammonium dimethanesulfonate Dichloromethane and 4,6-dimethoxy-1,3,5-triazin-2-yl methanesulfonate were added to a stirred reactor that had been fully replaced with N2. The temperature was lowered to 0°C and controlled at 0-5°C. Tetramethylethylenediamine was added dropwise, and the mixture was stirred for 1 hour. After the reaction was completed, the system was concentrated to 20% of its original volume and the temperature was controlled at 35-40°C. Then, methyl tert-butyl ether was added and the mixture was stirred and filtered. The mixture was dried at 35-40°C to obtain N,N-bis(4,6-dimethoxy-1,3,5-triazin-2-yl)-N,N,N,N-tetramethylethane-1,2-diammonium dimethanesulfonate, which is the condensing agent. The ratio of dichloromethane: 4,6-dimethoxy-1,3,5-triazine-2-ylmethanesulfonate: tetramethylethylenediamine: methyl tert-butyl ether in mL: g: g: ml is 50:5:1.2:

50.

4. A method for preparing the cosmetic peptide prolyl-prolyl-lysine using the condensing agent as described in claim 1, characterized in that: The method uses proline as a starting material, which reacts with benzyl bromide to generate proline with a benzyl-protected amino group. Then, in water, sodium bicarbonate is used to form a sodium salt and a condensing agent to form an active lipid, which directly reacts with unprotected proline. After the reaction is complete, Bn-Pro-Pro-OH is obtained. Then, Bn-Pro-Pro-OH is used in water to form a sodium salt with sodium bicarbonate, and a condensing agent is used to form an active lipid. Then, it is condensed with N6-Cbz-L-lysine to obtain Bn-Pro-Pro-Lys(Z)-OH. Finally, hydrogenation is performed to obtain the beauty peptide prolyl-prolyl-lysine.

5. The preparation method according to claim 4, characterized in that: The structural formula of the cosmetic peptide prolyl-prolyl-lysine is shown below: 。 6. The preparation method according to claim 4 or 5, characterized in that: Includes the following steps: (1) Preparation of Bn-Pro-OH Water and proline were added to a stirred reactor that had been fully replaced with N2. The temperature was lowered to 10°C, sodium carbonate was added, and the reaction was stirred for 30 min. Then benzyl bromide was added dropwise, and the reaction was stirred for 12 hours until the reaction was complete. The pH of the system was adjusted to 6.5-7.0 with 6N hydrochloric acid, filtered, and dried at 40-45°C to obtain Bn-Pro-OH. (2) Preparation of Bn-Pro-Pro-OH Add water and Bn-Pro-OH to a stirred reactor, slowly add sodium bicarbonate, control the temperature to 0-5℃ and add condensing agent, stir and react for 1 hour, add proline, stir and react for 2 hours. After the reaction is complete, adjust the pH to 6.5-7.0 with 6N hydrochloric acid, filter, and dry at 30-35℃ to obtain Bn-Pro-Pro-OH. (3) Preparation of Bn-Pro-Pro-Lys(Z)-OH Water and Bn-Pro-Pro-OH were added to a stirred reactor, sodium bicarbonate was slowly added, the temperature was controlled at 0-5℃, a condensing agent was added, and the reaction was stirred for 1 hour. H-Lys(Z)-OH was added, and the reaction was stirred for 2 hours. After the reaction was completed, the pH was adjusted to 6.5-7.0 with 6N hydrochloric acid, filtered, and dried at 30-35℃ to obtain Bn-Pro-Pro-Lys(Z)-OH. (4) Preparation of H-Pro-Pro-Lys-OH Purified water and Bn-Pro-Pro-Lys(Z)-OH were added to a stirred reactor that had been fully purged with N2. The temperature was controlled at 20-25℃, and Pd / C with a palladium content of 10% was added. The reactor was then purged with hydrogen three times and reacted under hydrogen protection. The temperature was controlled at 20-25℃ for 24 hours. After filtration, the system was concentrated to 20% of its original volume, and the temperature was controlled at 35-40℃. Ethanol was then added and the mixture was stirred. The mixture was filtered and dried at 30-35℃ to obtain H-Pro-Pro-Lys-OH, which yielded the beauty peptide prolyl-prolyl-lysine.

7. The preparation method according to claim 6, characterized in that: In step (1), the ratio of water:proline:sodium carbonate:benzyl bromide (mL:g:g:g) is 50:5:4.6:7.

4. In step (2), the ratio of water:Bn-Pro-OH:sodium bicarbonate:condensing agent:proline in mL:g:g:g:g is 50:3.5:2.8:5:1.

9. In step (3), the ratio of water:Bn-Pro-Pro-OH:sodium bicarbonate:condensing agent:H-Lys(Z)-OH in mL:g:g:g:g is 50:3:1.7:2.9:2.

7. In step (4), the ratio of purified water: Bn-Pro-Pro-Lys(Z)-OH: Pd / C: ethanol (ml: g: g: ml) is 50:2:0.2:

100.

8. The application of the preparation method according to any one of claims 4 to 7 in the preparation of the cosmetic peptide prolyl-prolyl-lysine.

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