N-terminal 1-position acetylation modified derivative of NPFF as well as preparation method and application of N-terminal 1-position acetylation modified derivative

Abstract

The invention discloses an N-terminal 1-position acetylation modified derivative of NPFF as well as a preparation method and application of the N-terminal 1-position acetylation modified derivative, and relates to the technical field of biological medicines. The method comprises the following steps: modifying an N terminal of neuropeptide FF (NPFF) by adopting acetylation of NPFF (N terminal 1 position) to generate an NPFF-Acety compound; the derivative provided by the invention shows remarkable anti-inflammatory activity in a mouse foot swelling inflammation model.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to an acetylated derivative of NPFF at the N-terminal 1 position, and a preparation method and application thereof. Background Art

[0002] Neuropeptide FF (NPFF) is a neuropeptide that regulates the body's quality of life in mammals and is widely distributed in multiple species. NPFF is a neuropeptide composed of eight amino acid residues, and its sequence is FLFQPQRFamide. NPFF has the function of regulating a variety of physiological activities, including pain, eating, obesity, lipid metabolism, endocrine, body temperature, blood pressure, emotions, inflammatory response, etc. The endogenous receptor of NPFF is generally considered to be NPFFR2 (NPFF receptor2, NPFFR2).

[0003] Although NPFF has good physiological activity, its application in the biomedical field is also subject to certain limitations, especially the inherent lack of stability of the polypeptide and the short half-life in vivo, which limit the application of such polypeptides.

[0004] Acetylation is a common method for modifying polypeptide residues. Acetylation of polypeptides is beneficial to enhancing the stability of polypeptides and improving their biological activity. In the process of acetylation of NPFF polypeptide, the 1st position of the N-terminus of NPFF, referred to as NPFF (1st position of the N-terminus), acetylation of NPFF (1st position of the N-terminus) can help change the biological activity and bring about possible therapeutic applications, thereby improving the therapeutic effect. So far, no method of modifying NPFF by acetylation has been seen. Therefore, acetylation of NPFF is expected to improve the stability of NPFF and enhance its biological activity, which is a new direction worth exploring in the research of polypeptide drugs. Summary of the invention

[0005] In view of the deficiencies in the above-mentioned background technology, the present invention provides a derivative of NPFF modified by acetylation at position 1 of the N-terminus, and a preparation method and application thereof. The method uses NPFF (position 1 of the N-terminus) to modify the N-terminus of neuropeptide FF (NPFF) to generate an NPFF-Acety derivative. The derivative showed significant anti-inflammatory activity in a mouse foot swelling inflammation model.

[0006] The first object of the present invention is to provide a method for preparing a derivative of NPFF with acetylation modification at position 1 of the N-terminus, comprising the following steps:

[0007] S1. Place the wang resin in N,N-dimethylamide to swell;

[0008] S2, dissolving the swollen resin, Fmoc-protected Phe, 4-dimethylaminopyridine, and N,N'-diisopropylcarbodiimide in DMF for reaction, removing unreacted reagents after the reaction, and then adding pyridine and acetic anhydride for end-capping treatment to obtain an end-capped compound;

[0009] S3, adding the capped compound to a 19% by volume piperidine DMF solution, then draining, and adding a 19% by volume piperidine DMF solution again to obtain a protected amino acid through reaction; then dissolving the protected amino acid and o-benzotriazole-tetramethyluronium hexafluorophosphate in DMF, and adding 4-methylmorpholine to carry out a condensation reaction;

[0010] Repeat S2 to S3, and condense the amino acids of the following sequence in order from the C-terminal to the N-terminal residue sequence: Acety-Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH2 to obtain a condensate;

[0011] The condensate, NH2-(CH2CH2O)8-CH2CH2COOH and HBTU were dissolved in DMF, and then NMM was added to react to remove the Fmoc protecting group, and then cleaved to obtain a cleavage solution containing the peptide;

[0012] The lysate was post-processed to obtain the N-terminal 1-acetylated derivative of NPFF.

[0013] Preferably, the lysis process comprises:

[0014] Prepare lysis solution: add ultrapure water, triisopropylsilane and 1,2-ethanedithiol to trifluoroacetic acid and mix well; wherein the volume ratio of trifluoroacetic acid, ultrapure water, triisopropylsilane and 1,2-ethanedithiol is 94:2:1:3;

[0015] The product after removal of the Fmoc protecting group is immersed in the lysis buffer for 110 to 220 minutes.

[0016] Preferably, after the lysis, the method further comprises: drying the lysate in a chlorine environment; washing the dried sample with ether for multiple times, and volatilizing the dried sample at room temperature until it is completely dry; and then purifying the dried sample with high performance liquid chromatography;

[0017] During purification, the mobile phase was a mixture of 0.16% trifluoroacetic acid and acetonitrile in a volume ratio of 50:50.

[0018] Preferably, the obtained end-capping compound, the product after condensation reaction and the product after removal of Fmoc protecting group are all washed in DMF and methanol for multiple cycles to remove impurities.

[0019] The second object of the present invention is to provide a derivative of NPFF with acetylation modification at position 1 of the N-terminus, which is Phe acetylated at the amino terminus of NPFF, and the amino acid sequence of Phe acetylated at the amino terminus of NPFF is: Acety-Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH2.

[0020] Preferably, the chemical structural formula of the derivative is as follows:

[0021]

[0022] The third object of the present invention is to provide an N-terminal 1-acetylated derivative of NPFF for use in the preparation of analgesics or anti-inflammatory drugs, and the derivative is prepared into tablets, capsules or injections according to conventional pharmacy.

[0023] The fourth object of the present invention is to provide an anti-inflammatory tablet comprising the following components in parts by weight: 40.0 parts of NPFF-Acety, 175.0 parts of a filler, 10.0 parts of a disintegrant, 5.0 parts of a binder, and 3.0 parts of a lubricant;

[0024] Among them, NPFF-Acety is a derivative of NPFF with acetylation modification at the N-terminal position 1.

[0025] The fifth object of the present invention is to provide an anti-inflammatory soft capsule, comprising the following components in parts by weight: 85.0 parts of NPFF-Acety, 85.0 parts of a filler, 5.0 parts of a binder, and 5.0 parts of a lubricant;

[0026] Among them, NPFF-Acety is a derivative of NPFF with acetylation modification at the N-terminal position 1.

[0027] The sixth object of the present invention is to provide an anti-inflammatory injection, comprising the following components by weight: 120.0 parts of NPFF-Acety, 1.0 parts of citric acid, 1.0 parts of sodium citrate, 16.0 parts of sodium chloride, and 2000 parts of water for injection;

[0028] Among them, NPFF-Acety is a derivative of NPFF with acetylation modification at the N-terminal position 1.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] The present invention provides an acetylated derivative of NPFF at position 1 of the N-terminus and a preparation method thereof. The present invention uses acetylation of NPFF (position 1 of the N-terminus) to modify the N-terminus of neuropeptide FF (NPFF) to generate an NPFF-Acety complex.

[0031] The present invention evaluates the anti-inflammatory activity of NPFF (N-terminal position 1) acetylation by a designed experimental scheme using a mouse foot swelling inflammation model. Experimental data show that NPFF (N-terminal position 1) acetylation exhibits significant anti-inflammatory properties. In a mouse foot swelling model simulating an inflammatory environment, 14 mg / kg and 28 mg / kg doses of NPFF (N-terminal position 1) acetylation were administered, which effectively inhibited the inflammatory response by 31.34% and 44.50% (after an observation period of 6 hours), respectively. This significant effect shows the therapeutic value of NPFF (N-terminal position 1) acetylation in controlling inflammation. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The mouse foot swelling inflammation model was used to evaluate the anti-inflammatory activity of NPFF (N-terminal position 1). VSControl, *P<0.05, **P<0.001. NPFF (N-terminal position 1) acetylation has significant anti-inflammatory activity. DETAILED DESCRIPTION

[0033] In order to enable those skilled in the art to better understand and implement the technical solution of the present invention, the present invention is further described below in conjunction with specific embodiments and drawings, but the embodiments are not intended to limit the present invention.

[0034] The first aspect of the present invention provides a method for preparing an N-terminal 1-acetylated derivative of NPFF, comprising the following steps:

[0035] S1. Place the wang resin in N,N-dimethylamide to swell;

[0036] S2, dissolving the swollen resin, Fmoc-protected Phe, 4-dimethylaminopyridine, and N,N'-diisopropylcarbodiimide in DMF for reaction, removing unreacted reagents after the reaction, and then adding pyridine and acetic anhydride for end-capping treatment to obtain an end-capped compound;

[0037] S3, adding the capped compound to a 19% by volume piperidine DMF solution, then draining, and adding a 19% by volume piperidine DMF solution again to obtain a protected amino acid through reaction; then dissolving the protected amino acid and o-benzotriazole-tetramethyluronium hexafluorophosphate in DMF, and adding 4-methylmorpholine to carry out a condensation reaction;

[0038] Repeat S2 to S3, and condense the amino acids of the following sequence in order from the C-terminal to the N-terminal residue sequence: Acety-Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH2 to obtain a condensate;

[0039] The condensate, NH2-(CH2CH2O)8-CH2CH2COOH and HBTU were dissolved in DMF, and then NMM was added to react to remove the Fmoc protecting group, and then cleaved to obtain a cleavage solution containing the peptide;

[0040] The lysate was post-processed to obtain the N-terminal 1-acetylated derivative of NPFF.

[0041] The cracking process includes:

[0042] Prepare lysis solution: add ultrapure water, triisopropylsilane and 1,2-ethanedithiol to trifluoroacetic acid and mix well; wherein the volume ratio of trifluoroacetic acid, ultrapure water, triisopropylsilane and 1,2-ethanedithiol is 94:2:1:3;

[0043] The product after removal of the Fmoc protecting group is immersed in the lysis buffer for 110 to 220 minutes.

[0044] After the lysis, the method further includes: drying the lysis solution in a chlorine environment; washing the dried sample with ether for multiple times, and volatilizing the dried sample at room temperature until it is completely dry; and then purifying the dried sample with high performance liquid chromatography;

[0045] During purification, the mobile phase was a mixture of 0.16% trifluoroacetic acid and acetonitrile in a volume ratio of 50:50.

[0046] The obtained end-capping compound, the product after the condensation reaction and the product after the Fmoc protecting group is removed are all washed in DMF and methanol for multiple cycles to remove impurities.

[0047] The second aspect of the present invention provides a derivative of NPFF with acetylation modification at position 1 of the N-terminus, wherein the derivative is Phe acetylated at the amino terminus of NPFF, and the amino acid sequence of Phe acetylated at the amino terminus of NPFF is: Acety-Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH2.

[0048] The chemical structure of the derivative is as follows:

[0049]

[0050] The third aspect of the present invention provides an application of an acetylated derivative of NPFF at position 1 of the N-terminus in the preparation of analgesics or anti-inflammatory drugs. The derivative is prepared into tablets, capsules or injections according to conventional pharmacy.

[0051] The fourth aspect of the present invention provides an anti-inflammatory tablet comprising the following components in parts by weight: 40.0 parts of NPFF-Acety, 175.0 parts of a filler, 10.0 parts of a disintegrant, 5.0 parts of a binder, and 3.0 parts of a lubricant;

[0052] Among them, NPFF-Acety is a derivative of NPFF with acetylation modification at the N-terminal position 1.

[0053] The fifth aspect of the present invention provides an anti-inflammatory soft capsule, comprising the following components in parts by weight: 85.0 parts of NPFF-Acety, 85.0 parts of a filler, 5.0 parts of a binder, and 5.0 parts of a lubricant;

[0054] Among them, NPFF-Acety is a derivative of NPFF with acetylation modification at the N-terminal position 1.

[0055] The sixth aspect of the present invention provides an anti-inflammatory injection, comprising the following components in parts by weight: 120.0 parts of NPFF-Acety, 1.0 parts of citric acid, 1.0 parts of sodium citrate, 16.0 parts of sodium chloride, and 2000 parts of water for injection;

[0056] Among them, NPFF-Acety is a derivative of the NPFF provided above with acetylation modification at position 1 of the N-terminus.

[0057] The present invention adopts NPFF (N-terminal 1 position) acetylation to modify the N-terminus of neuropeptide FF (NPFF) to generate NPFF-Acety derivatives. The derivatives exhibited significant anti-inflammatory activity in a mouse foot swelling inflammation model. The molecular formula of the compound is C 55 H 76 N 14 O 12 , molecular weight 1125.30.

[0058] The basic principle of using the mouse foot swelling inflammation model as a biological platform for evaluating in vivo anti-inflammatory activity is that carrageenan is used as a standard inflammatory factor and injected subcutaneously into the plantar surface of the hind limb of the mouse to induce an inflammatory response, causing an increase in exudate at the site of inflammation, which in turn increases the weight of the injected foot. By accurately weighing and comparing the weight difference between the inflamed and control sides of the foot, the degree of inflammation and the actual efficacy of the substance to be tested can be quantified. This method is a reliable and sensitive means of evaluating the in vivo activity of anti-inflammatory drugs.

[0059] Inhibition rate = (foot swelling value of the drug group - foot swelling value of the blank group) / (foot swelling value of the blank group) × 100%

[0060] The present invention verifies the anti-inflammatory effect of NPFF (position 1 of N-terminal) by acetylation and mouse paw inflammation. The experimental results show that acetylation of NPFF (position 1 of N-terminal) exhibits significant anti-inflammatory activity in the inflammatory model: in the paw edema model, the inflammation is reduced by 31.34% and 44.50% (12 hours) at doses of 14 mg / kg and 28 mg / kg, respectively. This discovery shows the application prospect of acetylation of NPFF (position 1 of N-terminal) as an anti-inflammatory drug, which has important medical value.

[0061] The acetylated NPFF (N-terminal position 1) compound refers to a derivative of NPFF modified by acetylation at the N-terminal position 1.

[0062] It should be noted that the experimental methods used in the present invention are all conventional methods unless otherwise specified; the reagents and materials used are all commercially available unless otherwise specified.

[0063] Example 1

[0064] The preparation of acetylated NPFF (position 1 at the N-terminus), specifically a method for preparing a derivative of acetylated NPFF at position 1 at the N-terminus, comprises the following steps:

[0065] Synthesis method: TBTU+DIEA+protected amino acid, all triple feed, modification on the amino group of Phe at the N-terminus.

[0066] The specific embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings. For method steps in the embodiments where specific experimental conditions are not explicitly mentioned, the standard operating procedures recommended by the manufacturer or conventional conditions generally recognized in the industry will be followed for execution.

[0067] The solid phase peptide synthesis method was used to prepare the acetylated derivative of NPFF (N-terminal position 1), and the steps were as follows:

[0068] Step 1: Resin swelling. Add N,N-dimethylformamide (DMF) to the reaction vessel, then add Wang resin, and shake for 21 minutes to allow the resin to swell fully.

[0069] Step 2: At room temperature, remove the solvent from the previous step by sand core filtration, add 1mmol and 5 times molar excess of Phe, and at the same time add 5 times molar excess of 4-dimethylaminopyridine and 5 times molar excess of N,N'-diisopropylcarbodiimide, and react at room temperature for 3 hours with DMF as solvent. After the reaction is completed, wash with DMF 5-8 times, 7-9mL each time, to remove unreacted reagents, etc. Then add appropriate amount of pyridine and acetic anhydride in a volume ratio of 1:1, react for 30 minutes, and obtain a capped compound.

[0070] Step 3: Deprotection: After removing DMF, add a 19% piperidine DMF solution by volume, drain for 7 minutes, add a 19% piperidine DMF solution by volume again, and react for 21 minutes.

[0071] Step 4: Detection. After removing the piperidine DMF solution, carefully pick up a small amount of resin particles with a sterile pipette and place them in a blank test tube, then add one drop each of ninhydrin, potassium cyanide and phenol solution. Heat the test tube at 101-110℃ for 5 minutes. If the solution turns dark blue, it indicates a positive result.

[0072] Step 5: Washing: Wash three times with DMF, methanol, and DMF in sequence to ensure that impurities are removed.

[0073] Step 6: Condensation reaction. The protected amino acid (4 times the molar amount of the synthetic peptide) and o-benzotriazole-tetramethyluronium hexafluorophosphate (HBTU) were dissolved in DMF and added to the reaction vessel. Subsequently, 4-methylmorpholine (NMM) (11 times the molar amount of the peptide) was quickly added to carry out the condensation reaction for 35 minutes.

[0074] Step 7: Washing: Wash twice with DMF (11 ml / g), then wash four times with methanol (11 ml / g), and finally wash twice with DMF (11 ml / g) to ensure that impurities are fully removed.

[0075] Step 8: Cyclic operation. Repeat the process from step 2 to step 7, and condense the following sequence of amino acids in the order from the C-terminal to the N-terminal residue:

[0076] Acety-Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH2

[0077] Step 9: Add NH2-(CH2CH2O)8-CH2CH2COOH (3.8 times the molar amount of NPFF) and HBTU (6 times the molar amount of peptide), both dissolved in DMF, then add NMM (19 times the molar amount of NPFF), and react for 35 minutes. Using the detection method in step 4, the result shows colorless, indicating that the Fmoc protecting group has been successfully removed.

[0078] Step 10: Perform multiple solvent washing and draining treatments. First, use DMF (10 ml / g) to wash three times, then use methanol (10 ml / g) to wash three times, then use DMF (10 ml / g) to wash three times, and finally use DMF (10 ml / g) to wash three times. After all washings are completed, drain the solution for 35 minutes.

[0079] Step 11: Cut the peptide on the resin. Specifically, mix 93% trifluoroacetic acid (V / V) with 8 ml per gram of resin as the cutting solution, and then add 3% ultrapure water (V / V), 1% triisopropylsilane (V / V) and 3.5% 1,2-ethanedithiol (V / V). Then, soak the resin with the cutting solution for 155 minutes.

[0080] Step 12: Blow dry and wash. Blow dry the lysate in a chlorine environment. Then, wash the dried sample nine times with ether and evaporate it naturally at room temperature (25°C) until it is completely dry.

[0081] Step 13: Preparation of dry powdered peptide crude product. The washed sample was purified by high performance liquid chromatography, and the mobile phase was a mixture of 0.16% trifluoroacetic acid and acetonitrile in a volume ratio of 50:50. Through this step, the pure acetylated derivative of NPFF was successfully prepared, and its structural formula is as follows:

[0082]

[0083] From the above structural formula, it can be seen that the mass spectrometry molecular weight of acetylated NPFF (position 1 at the N-terminus) is 1125.30, which is consistent with the theoretically calculated molecular weight, indicating that the acetylation of NPFF (position 1 at the N-terminus) was correctly identified.

[0084] Example 2. The anti-inflammatory activity of the acetylated NPFF (position 1 at the N-terminus) provided in Example 1 was evaluated using a mouse paw edema inflammation model.

[0085] 1) Experimental animals: The experimental mice were raised in a clean environment, with the ambient temperature maintained between 22-25°C, the humidity controlled between 46% and 53%, and sufficient light from 7:00 to 19:00 daily. There were 5 mice in each cage, and they were free to eat and drink.

[0086] 2) Experimental groups: The experiment was divided into four groups: a blank control group (Vehicle), a positive control group (DEX, dose of 5 mg / kg), and two different doses of NPFF (N-terminal 1 position) acetylation groups (12 mg / kg and 24 mg / kg, respectively). Each group contained 9 mice to ensure the consistency of experimental conditions.

[0087] 3) Mouse foot swelling inflammation model: Healthy mice aged 6-8 weeks were selected and randomly divided into several groups, each containing 9 mice. Each group of mice received different types of intraperitoneal injections of drugs. After 28 minutes, 23 μl of 1% (W / V) carrageenan was injected into the subcutaneous tissue of the right toe of the mouse as an inflammatory factor, while an equal amount of 0.9% saline was subcutaneously injected into the left toe as a control. Six hours after the injection of carrageenan, the thickness of the left and right feet of the mouse was measured and recorded using a vernier caliper. By calculating the difference in thickness between the right and left feet, the swelling thickness (unit: mm) was obtained to evaluate the inhibitory effect of the drug on the inflammatory response.

[0088] exist Figure 1 In the mouse foot swelling inflammation model shown, the acetylated drug NPFF (N-terminal position 1) showed an inflammation inhibition effect of 31.34% at a dose of 12 mg / kg; and when the dose was increased to 24 mg / kg, the inflammation inhibition rate increased to 44.50% (6 hours).

[0089] Example 3. Preparation of tablets

[0090] Formula: 40.0 parts of NPFF-Acety, 178.0 parts of filler, 6.0 parts of disintegrant, 7.0 parts of binder, 4.0 parts of lubricant, totaling 235.0 g.

[0091] Process: 1000 tablets were prepared according to the conventional process for preparing tablets, each tablet containing 50 mg of NPFF-Acety.

[0092] Dosage and administration: Once a day, 3 tablets each time.

[0093] Example 4. Preparation of soft capsules

[0094] Formula: 84.0 parts of NPFF-Acety, 86.0 parts of filler, 6.0 parts of adhesive, 4.0 parts of lubricant, totaling 180.0 g.

[0095] Process: 1000 capsules are prepared according to the conventional process for preparing capsules, each capsule containing 50 mg of NPFF-Acety.

[0096] Dosage and administration: Once a day, 3 capsules each time.

[0097] Example 5. Preparation of injection

[0098] Formula: 110.0 parts of NPFF-Acety, 1.5 parts of citric acid, 1.5 parts of sodium citrate, 14.0 parts of sodium chloride, 2000 ml of water for injection.

[0099] Process: According to the conventional process for preparing injections, a total of 1000 injections of 2 ml were prepared, each containing 120 mg of NPFF-Acety.

[0100] Dosage and administration: Once a day, 2 sticks each time.

[0101] The fillers, disintegrants, binders, lubricants and other excipients in Examples 3 to 5 are all the most common excipients in pharmacy. NPFF-Acety is a derivative of NPFF provided in Example 1 with acetylation modification at position 1 of the N-terminus.

[0102] In summary, the present invention not only successfully prepared NPFF (N-terminal position 1) acetylated as a new anti-inflammatory drug candidate, but also verified its significant anti-inflammatory effect through rigorous experiments. The application of NPFF (N-terminal position 1) acetylated as an anti-inflammatory drug provides a new strategy and idea for the treatment of inflammatory diseases, indicating that it has broad development prospects and potential market value in the medical field.

[0103] The present invention describes preferred embodiments and their effects. However, those skilled in the art may make additional changes and modifications to these embodiments once they are aware of the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0104] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a derivative of NPFF with acetylation modification at position 1 of the N-terminus, characterized in that: The following steps are involved: S1. Place the wang resin in N,N-dimethylamide to swell; S2, dissolving the swollen resin, Fmoc-protected Phe, 4-dimethylaminopyridine, and N,N'-diisopropylcarbodiimide in DMF for reaction, removing unreacted reagents after the reaction, and then adding pyridine and acetic anhydride for end-capping treatment to obtain an end-capped compound; S3, adding the blocked compound to a 19% by volume piperidine DMF solution, then draining, and adding a 19% by volume piperidine DMF solution again to obtain a protected amino acid through reaction; Then, the protected amino acid and o-benzotriazole-tetramethyluronium hexafluorophosphate are dissolved in DMF, and 4-methylmorpholine is added to carry out a condensation reaction; Repeat S2 to S3, and condense the amino acids of the following sequence in order from the C-terminal to the N-terminal residue sequence: Acety-Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH2 to obtain a condensate; The condensate, NH2-(CH2CH2O)8-CH2CH2COOH and HBTU were dissolved in DMF, and then NMM was added to react to remove the Fmoc protecting group, and then cleaved to obtain a cleavage solution containing the peptide; The lysate was post-processed to obtain the N-terminal 1-acetylated derivative of NPFF.

2. The method for preparing the derivative of NPFF with acetylation modification at position 1 of the N-terminus according to claim 1, characterized in that: The lysis process includes: Prepare lysis solution: add ultrapure water, triisopropylsilane and 1,2-ethanedithiol to trifluoroacetic acid and mix well; wherein the volume ratio of trifluoroacetic acid, ultrapure water, triisopropylsilane and 1,2-ethanedithiol is 94:2:1:3; The product after removal of the Fmoc protecting group is immersed in the lysis buffer for 110 to 220 minutes.

3. The method for preparing the derivative of NPFF with acetylation modification at position 1 of the N-terminus according to claim 1, characterized in that: After the lysis, the method further includes: drying the lysis solution in a chlorine environment; washing the dried sample with ether for multiple times, and volatilizing the dried sample at room temperature until it is completely dry; and then purifying the dried sample with high performance liquid chromatography; During purification, the mobile phase was a mixture of 0.16% trifluoroacetic acid and acetonitrile in a volume ratio of 50:

50.

4. The method for preparing the derivative of NPFF with acetylation modification at position 1 of the N-terminus according to claim 1, characterized in that: The obtained end-capping compound, the product after the condensation reaction and the product after the Fmoc protecting group is removed are all washed in DMF and methanol for multiple cycles to remove impurities.

5. A derivative of NPFF with acetylation modification at position 1 of the N-terminus obtained by the method according to any one of claims 1 to 4, characterized in that: The derivative is Phe at the amino terminus of NPFF modified by acetylation, and the amino acid sequence of Phe at the amino terminus of NPFF modified by acetylation is: Acety-Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH2.

6. The derivative of NPFF modified by acetylation at position 1 of the N-terminus according to claim 5, characterized in that: The chemical structure of the derivative is as follows:

7. Use of the N-terminal 1-acetylated derivative of NPFF according to claim 5 or 6 in the preparation of analgesics or anti-inflammatory drugs, characterized in that: The derivative is prepared into tablets, capsules or injections according to conventional pharmacy.

8. An anti-inflammatory tablet, characterized in that: The composition comprises the following components in parts by weight: 40.0 parts of NPFF-Acety, 175.0 parts of filler, 10.0 parts of disintegrant, 5.0 parts of binder, and 3.0 parts of lubricant; Among them, NPFF-Acety is a derivative of NPFF provided in claim 5 or 6 with acetylation modification at position 1 of the N-terminus.

9. An anti-inflammatory soft capsule, characterized in that: The composition comprises the following components in parts by weight: 85.0 parts of NPFF-Acety, 85.0 parts of filler, 5.0 parts of adhesive, and 5.0 parts of lubricant; Among them, NPFF-Acety is a derivative of NPFF provided in claim 5 or 6 with acetylation modification at position 1 of the N-terminus.

10. An anti-inflammatory injection, characterized in that: The invention comprises the following components in parts by weight: 120.0 parts of NPFF-Acety, 1.0 parts of citric acid, 1.0 parts of sodium citrate, 16.0 parts of sodium chloride, and 2000 parts of water for injection; Among them, NPFF-Acety is a derivative of NPFF provided in claim 5 or 6 with acetylation modification at position 1 of the N-terminus.

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