CN116284218A - Method for synthesizing palmitoyl tripeptide-1 based on liquid phase carrier assistance - Google Patents

Abstract

The invention belongs to the technical field of drug synthesis, in particular to a method for assisting synthesis of palmitoyl tripeptide-1 based on a liquid phase carrier. First synthesize four liquid phase carriers TAG-A, TAG-B, TAG-C and TAG-D, and then condense them with Fmoc-Lys(Boc)-OH to obtain compound 4A-4D, and obtain compound 5A-4D after deprotection 5D; compound 5A-5D was condensed with Fmoc-His(Trt)-OH to obtain compound 6A-6D, deprotected to obtain compound 7A-7D, compound 7A-7D was condensed with Fmoc-protected glycine to obtain compound 8A-8D, deprotected The compound 9A-9D was obtained, and the compound 9A-9D was condensed with palmitic acid to obtain the fully protected peptide compound 10A-10D; finally, the palmitoyl tripeptide-1 compound was obtained by cleavage with trifluoroacetic acid.

Description

Method for assisted synthesis of palmitoyl tripeptide-1 based on liquid phase carrier

technical field

The invention belongs to the field of drug synthesis, in particular to a method for assisting synthesis of palmitoyl tripeptide-1 based on a liquid phase carrier.

Background technique

Palmitoyl tripeptide-1 (English name Pal-Tripeptide-1) is composed of three amino acids and one fatty acid. The amino acid sequence is glycine-histidine-lysine (GHK). Palmitic acid can be combined with The N-terminals of GHK are linked to form the peptide of interest. Palmitoyl tripeptide-1 is a special amino acid polypeptide that acts on the dermis to make the skin firmer, relieve wrinkles, promote skin metabolism, stimulate cell proliferation, and achieve wrinkle removal and whitening. Anti-aging, increase skin radiance effect.

The synthesis method of palmitoyl tripeptide-1 includes two methods of solid-phase peptide synthesis and liquid-phase peptide synthesis, but the synthesis of palmitoyl tripeptide-1 by the solid-phase method is relatively short, the solid phase carrier is expensive, and the amount of amino acids is large. , resulting in higher cost; the liquid-phase synthesis method uses less amino acid and has a high yield, but the existing liquid-phase synthesis method has a complex post-treatment process, a large amount of organic solvents are used, the cost is high and it is not environmentally friendly.

CN108218956A discloses a liquid phase synthesis method of palmitoyl tripeptide-1. For the segmental synthesis of palmitoyl tripeptide-1, the dipeptide fragment H-His-Lys(Z)-OBzl and another fragment Pal-Gly-OH in the palmitoyl tripeptide-1 sequence were synthesized respectively, and then the two The fully protected peptide of palmitoyl tripeptide was synthesized by the method of activated ester, and then the final product palmitoyl tripeptide-1 was obtained through hydrogenation reaction. When synthesizing Pal-Gly-OH, the residue of Pal-OH is difficult to quantitatively detect. The synthesis of this fragment needs to calculate the dosage according to the synthesis of another fragment. At the same time, the intermediate Pal-Gly-ONb has high activity and is not easy to store. It also makes it difficult to carry out large-scale production, and the final step of hydrogenation and hydrogenation is likely to have the risk of explosion, which is not suitable for large-scale production.

CN112409444A and CN114891063A all use Boc-Lys(Z)-OH and benzyl bromide to react earlier in the liquid-phase synthesis of disclosed palmitoyl tripeptide-1 to obtain Boc-Lys(Z)-OBzl, and then remove Boc to obtain H -Lys(Z)-OBzl; then condense with Boc-His-OH and then remove Boc to obtain H-His-Lys(Z)-OBzl; repeat the amino acid coupling steps, and couple Boc-Gly-OH sequentially according to the peptide sequence , Pal-OH, to get Pal-Gly-His-Lys(z)-OBzl; finally hydrogenation debenzylation to get Pal-Gly-His-Lys-OH. This method uses a variety of controlled reagents, which is not conducive to large-scale production, and the palladium carbon used in the final hydrogenation reaction is also very dangerous, prone to fire and explosion risks, and is not suitable for large-scale production.

The synthesis of palmitoyl tripeptide-1 is mostly carried out by using the strategy of Boc to protect the amino group, first obtain Boc-Lys(Z)-OBzl and then condense with Boc-His-OH, Pal-Gly dipeptide or connect glycine first and then connect The fully protected peptide of palmitoyl tripeptide-1 was obtained by palmitic acid, and finally hydrogenated and debenzylated to obtain palmitoyl tripeptide-1. However, in the existing synthetic routes, it is not difficult to find that the post-treatment purification process is relatively complicated, and the types of solvents used are large, and the amount of solvents is large, and some solvents are regulated solvents, which is not conducive to large-scale production. In the final deprotection of the fully protected peptide, palladium carbon and hydrogen gas are used, which also has potential safety hazards in production, and there is a risk of explosion and fire.

The liquid-phase carrier technology gradually developed in recent years combines the advantages of the solid-phase peptide synthesis method and the traditional liquid-phase peptide synthesis method. By adding a protective group carrier with a long hydrophobic alkyl chain to the C-terminus of the target polypeptide, each Each step reaction can be carried out in a halogenated solvent, and the reaction intermediate can be precipitated with a polar organic solvent. Compared with the traditional liquid-phase peptide synthesis method, the post-processing operation is simple, and the purity of the intermediate in each step is relatively high; compared with the solid-phase peptide synthesis method, the utilization rate of amino acids is improved and the cost is saved. However, the existing liquid phase carrier assists the synthesis of peptide drugs: ①: the product may have a little viscosity in the deprotection step, ②: the product may be difficult to filter during the sedimentation process of some steps because the product particles are fine The leakage of material in the production rate will be reduced.

Contents of the invention

The purpose of the present invention is to provide a method for assisting the synthesis of palmitoyl tripeptide-1 based on a liquid phase carrier to address the deficiencies of the prior art.

The present invention first uses 4,4-dihydroxybenzophenone (compound 1) as a raw material, reacts with 1-bromooctadecane through an alkylation reaction to obtain compound 2, and then reduces compound 2 by sodium borohydride to obtain a hydrophobic liquid Phase carrier compound 3 (TAG-A).

Wherein, the amount of 1-bromooctadecane is between 2.2 equivalents and 3 equivalents; the amount of sodium borohydride is between 1.0 equivalents and 3.0 equivalents.

Formula 1. Liquid Phase Carrier Synthesis Route 1

Formula 2. Synthetic Route 2 of Liquid Phase Carrier

Palmitoyl tripeptide-1 can also be synthesized by using compound 14, compound 17 or compound 19 as a liquid carrier.

Compound 12 was alkylated first to obtain compound 13, which was then reduced by sodium borohydride to obtain compound 14 (TAG-B).

Compound 14 was reacted with thionyl chloride to obtain compound 15, compound 15 was reacted by SN ₂ to obtain compound 16, and compound 16 was reduced by sodium borohydride to obtain compound 17 (TAG-C).

In addition, compound 15 can also be obtained by SN ₂ reaction to obtain compound 18, which is then reduced by sodium borohydride to obtain compound 19 (TAG-D).

The method for assisted synthesis of palmitoyl tripeptide-1 based on liquid phase carrier is: liquid phase carrier TAG-A, TAG-B, TAG-C or TAG-D (compound 3/compound 14/compound 17/compound 19) and Fmoc- Condensation of Lys(Boc)-OH gives Fmoc-Lys(Boc)-TAG (compound 4A-4D), deprotection gives NH ₂ -Lys(Boc)-TAG (compound 5A-5D); and Fmoc-His(Trt) -OH condensation gives Fmoc-His(Trt)-Lys(Boc)-TAG (compounds 6A-6D), deprotection gives NH ₂ -His(Trt)-Lys(Boc)-TAG (compounds 7A-7D), and Condensation of Fmoc-protected glycine gives Fmoc-Gly-His(Trt)-Lys(Boc)-TAG (Compounds 8A-8D), deprotection gives NH ₂ -Gly-His(Trt)-Lys(Boc)-TAG (Compound 9A -9D), and then condensed with palmitic acid to obtain the fully protected peptide Pal-Gly-His(Trt)-Lys(Boc)-TAG (compound 10A-10D). Finally, palmitoyl tripeptide-1 (compound 11) can be obtained by cleavage with trifluoroacetic acid.

Formula 3. Synthetic route of palmitoyl tripeptide-1

Further: the synthesis method of compound 4A-4D is: adding dichloromethane, liquid phase carrier (TAG-OH) (compound 3/compound 14/compound 17/compound 19), Fmoc-Lys(Boc)-OH to the flask once (1.1 equivalents-1.8 equivalents), N,N'-diisopropylcarbodiimide (1.1 equivalents-1.8 equivalents), 4-dimethylaminopyridine (0.1 equivalents-0.6 equivalents), after stirring for 3 hours, the two Chloromethane was concentrated, and methanol was added under ice-cooling to form a solid. Stirring was continued for 10 minutes, and the mixture was filtered to collect compound 4A-4D as solid powder.

The synthetic method of compound 5A-5D is: add dichloromethane, compound 4A-4D, 1,8-diazobispiro[5.4.0] undec-7-ene (1.0 equivalent to 2.5 equivalent ) after stirring at room temperature for 30 minutes, dichloromethane was concentrated, acetonitrile was added under ice-cooling, and a solid precipitated, and the stirring was continued for 10 minutes, and then filtered to collect solid powder compounds 5A-5D.

The synthesis method of compound 6A-6D is: add dichloromethane, Fmoc-His(Trt)-OH (1.1 equivalents-1.8 equivalents), N,N-diisopropylethylamine (1.1 equivalents-1.8 eq), N,N'-diisopropylcarbodiimide (1.1 eq-1.8 eq), 1-hydroxybenzotriazole, (1.1 eq-1.8 eq), after stirring for 10 minutes, compound 5A-5D was added Stirring was continued for 3 hours in the reaction liquid, then dichloromethane was concentrated, methanol was added under ice-cooling, and solids were precipitated, and stirring was continued for 10 minutes, and then filtered to collect solid powders 6A-6D.

The synthetic method of compound 7A-7D is: add dichloromethane, compound 6A-6D, 1,8-diazobispiro[5.4.0]undec-7-ene (1.0 equivalent- 2.5 equivalents) after stirring at room temperature for 30 minutes, dichloromethane was concentrated, acetonitrile was added under ice-cooling, and a solid was precipitated, then stirring was continued for 10 minutes, and then filtered to collect solid powder compounds 7A-7D.

The synthesis method of compound 8A-8D is: add dichloromethane, Fmoc-Gly-OH (1.1 equivalents-1.8 equivalents), N,N-diisopropylethylamine (1.1 equivalents-1.8 equivalents), N,N'-diisopropylcarbodiimide (1.1-1.8 equivalents), 1-hydroxybenzotriazole, (1.1-1.8 equivalents) were stirred for 10 minutes and then compounds 7A-7D were added to the reaction solution Stirring was continued for 3 hours, then dichloromethane was concentrated, methanol was added under ice-cooling, and solids were precipitated, then stirring was continued for 10 minutes, and then filtered to collect solid powder compounds 8A-8D.

The synthesis method of compound 9A-9D is: add dichloromethane, compound 8A-8D, 1,8-diazobispiro[5.4.0]undec-7-ene (1.0 equivalent- 2.5 equivalents) after stirring at room temperature for 30 minutes, dichloromethane was concentrated, acetonitrile was added under ice-cooling, and a solid was precipitated, then stirring was continued for 10 minutes, and then filtered to collect solid powder compounds 9A-9D.

The synthesis method of compound 10A-10D is as follows: add dichloromethane, palmitic acid (1.1 equivalent to 1.8 equivalent), N,N-diisopropylethylamine (1.1 equivalent to 1.8 equivalent), N,N' -Diisocarbodiimide (1.1 equivalents-1.8 equivalents), 1-hydroxybenzotriazole, (1.1 equivalents-1.8 equivalents) were stirred for 10 minutes and then compound 9A-9D was added to the reaction solution and continued to stir for 3 hours , then dichloromethane was concentrated, methanol was added under ice bath, and solid was precipitated, then stirred for 10 minutes, filtered to collect solid powder, and dried in a vacuum oven at 30°C to obtain white solid compounds 10A-10D.

The synthesis method of palmitoyl tripeptide-1 is as follows: add dichloromethane and compound 10A-10D in turn to the round bottom flask, add cutting solution after the reaction solution dissolves and continue to stir for 3 hours, concentrate the reaction solution, add iso Continue to stir for 30 minutes after the propyl ether has solid precipitation, then filter, collect the filter cake, stir and dissolve the filter cake with 100 ml of water and filter, prepare and purify the filtrate, and freeze-dry the collected solution to obtain the white solid final product palm Acyl tripeptide-1.

Beneficial effect:

The present invention utilizes a liquid phase carrier to synthesize palmitoyl tripeptide-1, uses dichloromethane as a solvent, and has mild reaction conditions and simple post-treatment process. After the reaction, the product can be precipitated by a highly polar solvent, methanol or acetonitrile, and is suitable for large-scale production. .

The carrier of the invention is soluble, and the reaction is a homogeneous reaction; the utilization rate of the amino acid is improved, and the cost is saved.

Description of drawings:

Figure 1: Ms diagram of palmitoyl tripeptide-1.

Detailed ways

In order to make the purpose, technical solutions and advantages of the present invention clearer, the present invention will be further explained below in conjunction with specific embodiments, but it is not intended to limit the protection scope of the present invention.

Example 1

At 25±5°C, 150 ml of N,N-dimethylformamide, compound 1 (10 g, 46.68 mmol), potassium carbonate (14.2 g, 102.7 mmol), 1 -Bromooctadecane (34.4 g, 102.7 mmol), then reacted at 90° C. for 4 hours. After the reaction is completed, pour it into 750ml of water while it is still hot, stir at 25±5°C for 30 minutes and then filter, beat the filter cake with 500ml of methanol and filter, collect the white solid and dry it in a blast drying oven at 45°C to obtain a white solid Compound 2 32.6 g, yield 97%.

At 25±5°C, add 350 ml of tetrahydrofuran, compound 2, and 35 ml of methanol to a 1-liter round-bottomed flask in sequence, then heat at 60°C, and add sodium borohydride (1.7 g, 45 mmol) after the reaction liquid is dissolved to react 2 Hour. After the reaction is finished, wait for the reaction solution to drop to room temperature, slowly add water dropwise to the reaction solution, and pour the reaction solution into 2 liters of water and stir for 30 minutes before filtering when there are no obvious bubbles in the reaction solution. The filter cake is beaten with 500 ml of methanol and then filtered. , collected the white solid and dried it in a forced air oven at 45° C. to obtain 31.3 g of white solid compound 3 (TAG-A), with a yield of 96%.

At 25±5°C, add 50 ml of dichloromethane, compound 3 (TAG-A, 5 g, 6.93 mmol), Fmoc-Lys(Boc)-OH (commercially available, Zhengyuan, Chengdu) into a 250 ml round bottom flask. ) (3.9 g, 8.32 mmol), N,N'-diisopropylcarbodiimide (1.6 mL, 10.4 mmol), 4-dimethylaminopyridine (169 mg, 1.387 mmol) after stirring for 3 hours , dichloromethane was concentrated to about 5 milliliters, and 100 milliliters of methanol was added under ice-cooling, and the stirring was continued for 10 minutes after the solid precipitated out. Then filter, collect the solid powder compound 4A, drain it and put it into the next step.

At 25±5°C, add 50 ml of dichloromethane, compound 4A, 1,8-diazobispiro[5.4.0]undec-7-ene (1.036 ml, 6.93 mmol) and stirred at room temperature for 30 minutes, dichloromethane was concentrated to about 5 milliliters, and 100 milliliters of acetonitrile was added under ice-cooling, and a solid was precipitated, and then the stirring was continued for 10 minutes. Then filter, collect the solid powder compound 5A, suck it dry and throw it into the next step.

At 15±5°C, add 50 ml of dichloromethane, Fmoc-His(Trt)-OH (5.157 g, 8.32 mmol) (commercially available, Chengdu Zhengyuan), N,N-Di Isopropylethylamine (1.56ml, 9mmol), N,N'-diisopropylcarbodiimide (1.6ml, 10.4mmol), 1-hydroxybenzotriazole, (1.4g, 10.4mM mol) was stirred for 10 minutes, and then compound 5A was added into the reaction solution and stirred for 3 hours. Then dichloromethane was concentrated to about 5 milliliters, 100 milliliters of methanol was added under ice-cooling, solid was precipitated, and stirring was continued for 10 minutes. Then filter, collect the solid powder compound 6A, suck it dry and put it into the next step.

At 25±5°C, add 50 ml of dichloromethane, compound 6A, 1,8-diazobispiro[5.4.0]undec-7-ene (1.036 ml, 6.93 mmol) and stirred at room temperature for 30 minutes, dichloromethane was concentrated to about 5 milliliters, and 100 milliliters of acetonitrile was added under ice-cooling, and a solid was precipitated, and then the stirring was continued for 10 minutes. Then filter, collect the solid powder compound 7A, suck it dry and put it into the next step.

At 20±5°C, 75 ml of dichloromethane, Fmoc-Gly-OH (2.47 g, 8.32 mmol), N,N-diisopropylethylamine (1.56 ml, 9 mmol), N,N'-diisopropylcarbodiimide (1.6 ml, 10.4 mmol), 1-hydroxybenzotriazole, (1.4 g, 10.4 mmol) were stirred for 10 minutes and compound 7A Added to the reaction solution and continued to stir for 3 hours. Then dichloromethane was concentrated to about 5 milliliters, 100 milliliters of methanol was added under ice-cooling, solid was precipitated, and stirring was continued for 10 minutes. Then filter, collect the solid powder compound 8A, suck it dry and put it into the next step.

At 25±5°C, add 50 ml of dichloromethane, compound 8A, 1,8-diazobispiro[5.4.0]undec-7-ene (1.036 ml, 6.93 mmol) and stirred at room temperature for 30 minutes, dichloromethane was concentrated to about 5 milliliters, and 100 milliliters of acetonitrile was added under ice-cooling, and a solid was precipitated, and then the stirring was continued for 10 minutes. Then filter, collect the solid powder compound 9A, drain it and put it into the next step.

At 20±5°C, add 75 ml of dichloromethane, palmitic acid (2.13 g, 8.32 mmol), N,N-diisopropylethylamine (1.56 ml, 9 mmol) in a 250 ml round bottom flask , N, N'-diisocarbodiimide (1.6 milliliters, 10.4 mmol), 1-hydroxybenzotriazole, (1.4 grams, 10.4 mmol) and then compound 9A was added to the reaction solution after stirring for 10 minutes Stirring was continued for 3 hours. Then dichloromethane was concentrated to about 5 milliliters, 100 milliliters of methanol was added under ice-cooling, solid was precipitated, and stirring was continued for 10 minutes. Then filter, collect the solid powder, and dry in a vacuum oven at 30°C to obtain 10.55 g of white solid compound 10A, with a yield of 93.8%.

At 10±5°C, add 50 ml of dichloromethane and compound 10A in sequence to a 500 ml round bottom flask, and add 50 ml of cutting fluid (trifluoroacetic acid: triisopropylsilane: water = 95: 2.5:2.5) Stirring was continued for 3 hours. The reaction solution was concentrated to about 20 ml, and 200 ml of isopropyl ether was added at 0° C., and a solid was precipitated, and the stirring was continued for 30 minutes. Then filter, collect filter cake, filter cake is stirred and dissolved with 100 milliliters of water after filtering, and filtrate is prepared and purified. The collected solution was lyophilized with a lyophilizer to obtain a white solid final product with a yield of 62.5% and a purity of >98%.

Example 2

At 25±5°C, 150 ml of N,N-dimethylformamide, compound 12 (10 g, 72.46 mmol), potassium carbonate (30 g, 217 mmol), 1 -Bromooctadecane (53.14 g, 159 mmol), then reacted at 90° C. for 4 hours. After the reaction is completed, pour it into 750ml of water while it is still hot, stir at 25±5°C for 30 minutes and then filter, beat the filter cake with 500ml of methanol and filter, collect the white solid and dry it in a blast drying oven at 45°C to obtain a white solid Compound 13 45.57 g, yield 98%.

At 25±5°C, add 400ml of tetrahydrofuran, compound 13, and 45ml of methanol to a 1-liter round-bottomed flask successively, then heat at 60°C, and add sodium borohydride (2.68g, 71mmol) after the reaction liquid is dissolved to react 2 Hour. After the reaction is finished, wait for the reaction solution to drop to room temperature, slowly add water dropwise to the reaction solution, and pour the reaction solution into 2 liters of water and stir for 30 minutes before filtering when there are no obvious bubbles in the reaction solution. The filter cake is beaten with 500 ml of methanol and then filtered. , collected the white solid and dried it in a forced air oven at 45° C. to obtain 45 g of white solid compound 14 (TAG-B), with a yield of 98%.

An embodiment of the liquid-phase synthesis method of the present invention based on the liquid-phase carrier-assisted synthesis of palmitoyl tripeptide-1, except that the liquid-phase carrier used (compound 14 (TAG-B) is used in this example) is different from that of Example 1 Except, other steps are consistent with embodiment 1. Finally, the yield of palmitoyl tripeptide-1 was 53%.

Example 3

At 0±5°C, add 100 ml of dichloromethane, compound 14, and 1 ml of thionyl chloride to a 250 ml round-bottomed flask in sequence, and then react at 0°C for 1 hour. Methylene chloride was washed once with 5% aqueous sodium bicarbonate solution, then the dichloromethane was dried over anhydrous magnesium sulfate and concentrated to dryness to obtain 4.8 g of compound 15 with a yield of 93%.

At 25±5°C, 80 ml of N,N-dimethylformamide, compound 15 (4.8 g, 7.2 mmol), potassium carbonate (2 g, 14.5 mmol), and para Hydroxybenzaldehyde (1.06 g, 8.7 mmol) was then reacted at 80° C. for 4 hours. After the reaction is over, pour it into 500ml of water while it is hot, stir at 25±5°C for 30 minutes and then filter, beat the filter cake with 200ml of methanol and filter, collect the white solid and dry it in a blast drying oven at 45°C to obtain a white solid Compound 16 5.1 g, yield 94%.

At 25±5°C, add 45 ml of tetrahydrofuran, compound 16, and 5 ml of methanol to a 250 ml round-bottomed flask successively, then heat at 60°C, and add sodium borohydride (0.26 g, 6.8 mmol) after the reaction liquid is dissolved to react 2 Hour. After the reaction is finished, wait for the reaction solution to drop to room temperature, and slowly add water dropwise to the reaction solution. When there are no obvious bubbles in the reaction solution, pour the reaction solution into 300 ml of water and stir for 30 minutes before filtering. The filter cake is beaten with 100 ml of methanol and then filtered. , collected the white solid and dried it in a forced air oven at 45° C. to obtain 4.9 g of white solid compound 17 (TAG-C), with a yield of 96%.

An embodiment of the liquid-phase synthesis method of the present invention based on liquid-phase carrier-assisted synthesis of palmitoyl tripeptide-1, except that the liquid-phase carrier used is different from Example 1 (compound 17 (TAG-C) is used in this example) Except, other steps are consistent with embodiment 1. Finally, the yield of palmitoyl tripeptide-1 was 65%.

Example 4

At 25±5°C, 80 ml of N,N-dimethylformamide, compound 15 (4.8 g, 7.2 mmol), potassium carbonate (2 g, 14.5 mmol), 4 -(allyloxy)-2-hydroxybenzaldehyde (1.54 g, 8.7 mmol), then reacted at 80°C for 4 hours. After the reaction is over, pour it into 500ml of water while it is hot, stir at 25±5°C for 30 minutes and then filter, beat the filter cake with 200ml of methanol and filter, collect the white solid and dry it in a blast drying oven at 45°C to obtain a white solid Compound 18 5.3 g, yield 91%.

At 25±5°C, add 45 ml of tetrahydrofuran, compound 18, and 5 ml of methanol to a 250 ml round-bottomed flask in sequence, then heat to 60°C, and add sodium borohydride (0.25 g, 6.6 mmol) after the reaction liquid is dissolved to react 2 Hour. After the reaction is finished, wait for the reaction solution to drop to room temperature, and slowly add water dropwise to the reaction solution. When there are no obvious bubbles in the reaction solution, pour the reaction solution into 300 ml of water and stir for 30 minutes before filtering. The filter cake is beaten with 100 ml of methanol and then filtered. , collected the white solid and dried it in a forced air oven at 45° C. to obtain 4.7 g of white solid compound 19 (TAG-D), with a yield of 88%.

An embodiment of the liquid-phase synthesis method of the present invention based on the liquid-phase carrier-assisted synthesis of palmitoyl tripeptide-1, except that the liquid-phase carrier used is different from that of Example 2 (compound 19 (TAG-D) is used in this embodiment) Except, other steps are consistent with embodiment 2. Finally, the yield of palmitoyl tripeptide-1 was 63%.

The above embodiments are only used to illustrate the technical solution of the present invention, and the description thereof is specific and detailed, but not limiting. For those skilled in the art, various modifications and changes can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention.

Claims (10)

1. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier is characterized by comprising the following steps of: condensing four liquid phase carriers TAG-A, TAG-B, TAG-C or TAG-D with Fmoc-Lys (Boc) -OH respectively to obtain compounds 4A-4D, and deprotecting to obtain compounds 5A-5D; condensing the compound 5A-5D with Fmoc-His (Trt) -OH to obtain a compound 6A-6D, deprotecting to obtain a compound 7A-7D, condensing the compound 7A-7D with Fmoc-protected glycine to obtain a compound 8A-8D, deprotecting to obtain a compound 9A-9D, condensing the compound 9A-9D with palmitic acid to obtain a full-protected peptide compound 10A-10D; finally obtaining palmitoyl tripeptide-1 compound 11 through trifluoroacetic acid cleavage; the specific formula is as follows:

2. ext> theext> methodext> forext> synthesizingext> palmitoylext> tripeptideext> -ext> 1ext> basedext> onext> theext> liquidext> phaseext> carrierext> assistanceext> accordingext> toext> claimext> 1ext>,ext> whereinext> theext> synthesisext> methodext> ofext> TAGext> -ext> Aext> isext> asext> followsext>:ext> Ext> 4ext>,ext> 4ext> -ext> dihydroxybenzophenoneext> isext> usedext> asext> aext> rawext> materialext>,ext> theext> rawext> materialext> isext> reactedext> withext> 1ext> -ext> bromooctadecaneext> throughext> alkylationext> reactionext> toext> obtainext> aext> compoundext> 2ext>,ext> andext> theext> compoundext> 2ext> isext> reducedext> byext> sodiumext> borohydrideext> toext> obtainext> aext> hydrophobicext> liquidext> phaseext> carrierext> TAGext> -ext> Aext>,ext> whereinext> theext> specificext> formulaext> isext> asext> followsext>:ext>

The synthesis method of TAG-B comprises the following steps: the compound 12 is subjected to alkylation reaction to obtain a compound 13, and the compound 13 is reduced by sodium borohydride to obtain a liquid carrier TAG-B;

the synthesis method of TAG-C comprises the following steps: TAG-B under the action of thionyl chloride to obtain a compound 15, wherein the compound 15 is prepared by SN ₂ ReactionObtaining a compound 16, and reducing the compound 16 by sodium borohydride to obtain a liquid carrier TAG-C;

the synthesis method of TAG-D comprises the following steps: TAG-B under the action of thionyl chloride to obtain a compound 15, wherein the compound 15 is prepared by SN ₂ Obtaining a compound 18 through reaction, and reducing the compound 18 by sodium borohydride to obtain a liquid carrier TAG-D;

3. the method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 4A-4D is as follows: dichloromethane, a liquid carrier TAG-A, TAG-B, TAG-C or TAG-D, fmoc-Lys (Boc) -OH, N' -diisopropylcarbodiimide and 4-dimethylaminopyridine are sequentially added into a flask, the mixture is stirred for 3 hours, the dichloromethane is concentrated, methanol is added under ice bath, the mixture is stirred for 10 minutes after solid precipitation, and the solid powder compounds 4A-4D are filtered and collected.

4. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 5A-5D is as follows: dichloromethane, the compounds 4A-4D and 1, 8-diazabicyclo [5.4.0] undec-7-ene are sequentially added into a flask, stirred for 30 minutes at room temperature, then dichloromethane is concentrated, acetonitrile is added into the flask under ice bath, stirring is continued for 10 minutes after solid precipitation, then filtration is carried out, and solid powder compounds 5A-5D are collected.

5. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 6A-6D is as follows: dichloromethane, fmoc-His (Trt) -OH, N-diisopropylethylamine, N' -diisopropylcarbodiimide and 1-hydroxybenzotriazole are sequentially added into a round bottom flask, after stirring for 10 minutes, the compounds 5A-5D are added into a reaction liquid, stirring is continued for 3 hours, then dichloromethane is concentrated, methanol is added under ice bath, stirring is continued for 10 minutes after solid precipitation, and then filtration is carried out, and solid powder compounds 6A-6D are collected.

6. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 7A-7D is as follows: dichloromethane, the compound 6A-6D and 1, 8-diazabicyclo [5.4.0] undec-7-ene are sequentially added into a round bottom flask, stirred for 30 minutes at room temperature, then dichloromethane is concentrated, acetonitrile is added into the mixture under ice bath, stirring is continued for 10 minutes after solid precipitation, and the solid powder compound 7A-7D is filtered and collected.

7. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 8A-8D is as follows: dichloromethane, fmoc-Gly-OH, N-diisopropylethylamine, N' -diisopropylcarbodiimide and 1-hydroxybenzotriazole are sequentially added into a round bottom flask, after stirring for 10 minutes, the compounds 7A-7D are added into a reaction liquid, stirring is continued for 3 hours, then dichloromethane is concentrated, methanol is added under ice bath, stirring is continued for 10 minutes after solid precipitation, and then filtering is carried out, and solid powder compounds 8A-8D are collected.

8. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 9A to 9D is as follows: dichloromethane, the compounds 8A-8D and 1, 8-diazabicyclo [5.4.0] undec-7-ene are sequentially added into a round bottom flask, stirred for 30 minutes at room temperature, then dichloromethane is concentrated, acetonitrile is added into the round bottom flask under ice bath, stirring is continued for 10 minutes after solid precipitation, then filtration is carried out, and solid powder compounds 9A-9D are collected.

9. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 10A to 10D is as follows: adding dichloromethane, palmitic acid, N-diisopropylethylamine, N' -diisopropylethylamine and 1-hydroxybenzotriazole into a round bottom flask in sequence, stirring for 10 minutes, adding the compounds 9A-9D into a reaction liquid, continuously stirring for 3 hours, concentrating the dichloromethane, adding methanol under ice bath, continuously stirring for 10 minutes after solid precipitation, filtering, collecting solid powder, and drying in a vacuum drying oven at 30 ℃ to obtain white solid compounds 10A-10D.

10. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of palmitoyl tripeptide-1 is as follows: adding dichloromethane and compounds 10A-10D into a round bottom flask in sequence, adding cutting fluid after the reaction fluid is dissolved, continuously stirring for 3 hours, concentrating the reaction fluid, adding isopropyl ether at 0 ℃ to precipitate solids, continuously stirring for 30 minutes, filtering, collecting a filter cake, stirring and dissolving the filter cake with 100 milliliters of water, filtering, preparing and purifying the filtrate, and freeze-drying the collected fluid by a freeze dryer to obtain a white solid final product.

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