A COX2 inhibitory peptide, a composition for inhibiting COX2, and its application.

Abstract

This invention provides a COX2 inhibitory peptide, a composition for inhibiting COX2, and its application, belonging to the field of bioactive peptide technology. The amino acid sequence of the COX2 inhibitory peptide of this invention includes RGPF, FPK, or NPW. Studies have found that COX2 inhibitory peptides with amino acid sequences such as RGPF, FPK, or NPW exhibit an IC50 (increase in activity) for inhibiting COX2 activity. 50 The values ​​were 360.17±32.60 μM, 640.33±10.29 μM, and 1100.90±89.93 μM, respectively, which can effectively inhibit the activity of COX2, thereby effectively preventing and / or treating inflammation and cancer caused by COX2, and also achieving antipyretic and analgesic effects. Meanwhile, the COX2 inhibitory peptide of this invention has the advantages of being safe, non-toxic, and highly water-soluble, providing a safe option for the clinical use of COX2 inhibitors.

Description

Technical Field

[0001] This invention belongs to the field of bioactive peptide technology, and particularly relates to a COX2 inhibitory peptide, a composition for inhibiting COX2, and its application. Background Technology

[0002] Cyclooxygenase-2 (COX2) is an inducible cyclooxygenase, part of the COX enzyme class, which, along with COX1, participates in prostaglandin synthesis. COX2 is expressed in various tissues, but its expression is typically low in non-inflammatory sites. Growth factors, cytokines, and bacterial lipopolysaccharides can significantly increase COX2 expression, catalyzing the conversion of arachidonic acid into prostaglandins, thereby causing a series of adverse reactions such as inflammation, pain, and even cancer.

[0003] Studies have found that COX2 expression and enzyme activity are significantly elevated in patients with neurodegenerative diseases, rheumatoid arthritis, lupus erythematosus, irritable bowel syndrome, and cancer. The relationship between COX2 and inflammation and cancer makes it a potential biomarker and drug target. Using COX2 inhibitors can effectively alleviate symptoms of related diseases and, to some extent, influence disease progression.

[0004] Currently, COX2 inhibitors are mainly chemically synthesized nonsteroidal anti-inflammatory drugs (NSAIDs), including salicylates, aryl or heterocyclic aryl derivatives, and hydroxybenzothiazides (coxib derivatives). However, while these drugs have COX2 inhibitory activity, they also increase the risk of thrombotic cardiovascular disease. In 2005, the US FDA determined that the increased risk of cardiovascular events from coxib derivatives was unavoidable and recommended suspending the sale of vardicoxib.

[0005] As research has progressed, some natural compounds have shown the potential to inhibit COX2 activity. A large number of natural compounds, such as phenols, flavonoids, and alkaloids, have been identified as COX2 inhibitors or have exerted COX2 inhibitory activity through certain structural modifications. However, these compounds still have problems such as poor water solubility and low absorption rate.

[0006] Peptides have been shown in previous studies to possess various biological activities and high nutritional value. More importantly, peptides are non-toxic, safer, and easily digested and absorbed by the body. However, there are currently no reports on the use of COX2 inhibitory peptides in the preparation of COX2 inhibitory drugs or reagents. Summary of the Invention

[0007] Therefore, the purpose of this invention is to provide a COX2 inhibitory peptide, a composition for inhibiting COX2, and its application, wherein the COX2 inhibitory peptide has a COX2 inhibitory effect.

[0008] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0009] This invention provides a COX2 inhibitory peptide, wherein the amino acid sequence of the COX2 inhibitory peptide includes RGPF, FPK or NPW.

[0010] The present invention also provides a composition for inhibiting COX2, wherein the active ingredient of the composition comprises one or more of the above-mentioned RGPF, FPK and NPW.

[0011] Preferably, the composition further includes pharmaceutically acceptable excipients.

[0012] Preferably, the sole active ingredient of the composition is one or more of the above-mentioned RGPF, FPK and NPW.

[0013] The present invention also provides the use of the above-mentioned COX2 inhibitory peptide or composition in the preparation of COX2-inhibiting products.

[0014] The present invention also provides the use of the above-mentioned COX2 inhibitory peptide or composition in the preparation of products for the prevention and / or treatment of COX2-induced diseases.

[0015] The present invention also provides the use of the above-mentioned COX2 inhibitory peptide or composition in the preparation of products for the prevention and / or treatment of COX2-induced pain and / or fever.

[0016] Preferably, the disease includes one or both of inflammation and cancer.

[0017] Preferably, the product includes reagents or drugs.

[0018] Preferably, the dosage form of the product includes any pharmaceutically acceptable dosage form.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] This invention provides a COX2 inhibitory peptide, a composition for inhibiting COX2, and its applications. Through research, this invention has discovered that COX2 inhibitory peptides with amino acid sequences such as RGPF, FPK, or NPW exhibit an IC50 of inhibiting COX2 activity. 50 The values ​​were 360.17±32.60 μM, 640.33±10.29 μM, and 1100.90±89.93 μM, respectively, which can effectively inhibit the activity of COX2, thereby effectively preventing and / or treating inflammation and cancer caused by COX2, and also achieving antipyretic and analgesic effects. Meanwhile, the COX2 inhibitory peptide of this invention has the advantages of being safe, non-toxic, and highly water-soluble, providing a new option for the safe clinical use of COX2 inhibitors. Attached Figure Description

[0021] Figure 1 The inhibitory activity of RGPF active peptide, FPK active peptide, or NPW active peptide on COX2 and IC50 at different concentrations. 50 value. Detailed Implementation

[0022] This invention provides a COX2 inhibitory peptide, wherein the amino acid sequence of the COX2 inhibitory peptide includes RGPF, FPK or NPW.

[0023] The present invention also provides a composition for inhibiting COX2, wherein the active ingredient of the composition comprises one or more of the above-mentioned RGPF, FPK and NPW.

[0024] In this invention, the composition further includes pharmaceutically acceptable excipients. These excipients include excipients. Excipients broadly refer to any component other than the active ingredient, and can be inert, inactive, and / or pharmaceutically inactive substances. Excipients can serve various purposes, such as acting as carriers, solvents, diluents, tablet additives, and / or improving the release and absorption of the active ingredient, including one or more of solvents, diluents, buffers, preservatives, isotonic agents, pH adjusters, chelating agents, and stabilizers. Buffers include one or more of Krebs buffer, sodium dihydrogen phosphate, disodium hydrogen phosphate, succinate, maleic acid, fumaric acid, and tartaric acid. Preservatives include one or more of phenol, o-cresol, m-cresol, p-cresol, methylparaben, propylparaben, 2-phenoxyethanol, butylparaben, benzyl alcohol, benzoic acid, chlorocresol, and chlorophenoxyether. Chelating agents may be selected from ethylenediaminetetraacetic acid or citric acid. The isotonic agent may be selected from one or more of salts (e.g., sodium chloride), sugar alcohols (e.g., glycerol, propylene glycol, 1,3-propanediol, 1,3-butanediol), and polyethylene glycols (e.g., PEG400).

[0025] In this invention, the sole active ingredient of the composition is one or more of the above-mentioned RGPF, FPK and NPW.

[0026] The present invention also provides the use of the above-mentioned COX2 inhibitory peptide or composition in the preparation of COX2-inhibiting products.

[0027] The present invention also provides the use of the above-mentioned COX2 inhibitory peptide or composition in the preparation of products for the prevention and / or treatment of COX2-induced diseases.

[0028] The present invention also provides the use of the above-mentioned COX2 inhibitory peptide or composition in the preparation of products for the prevention and / or treatment of COX2-induced pain and / or fever.

[0029] As one possible implementation, the disease includes one or both of inflammation and cancer. The product includes a reagent or drug. The product can use a COX2 inhibitory peptide or composition as the sole active ingredient to inhibit COX2 activity, or it can be combined with other COX2-inhibiting ingredients to inhibit COX2 activity, thereby achieving the effects of preventing and / or treating inflammation and cancer, while also having antipyretic and analgesic effects. This invention has found that RGPF, FPK, or NPW active peptides can effectively inhibit COX2 activity, with an IC50 of [missing information - likely related to COX2 activity inhibition]. 50 The values ​​were 360.17±32.60μM, 640.33±10.29μM, and 1100.90±89.93μM, respectively.

[0030] In this invention, the dosage form of the product includes any pharmaceutically acceptable dosage form, such as sustained-release formulation, capsules, granules, oral liquid, tablets, or injections. The product can be administered orally or by injection. The COX2 inhibitory peptide or composition constitutes 50% to 100% of the product by mass, such as 50%, 60%, 70%, 80%, 90%, or 100%. The product also includes pharmaceutically acceptable excipients; the types of excipients are described in the section on pharmaceutically acceptable excipients mentioned in the above composition description and will not be repeated here.

[0031] In this invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0032] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0033] Example 1

[0034] Assay for the activity of COX2 inhibitory peptide

[0035] In this embodiment, the amino acid sequence of the COX2 inhibitory peptide is RGPF (SEQ ID NO.1), FPK, or NPW, and is named RGPF active peptide, FPK active peptide, or NPW active peptide, respectively.

[0036] The in vitro inhibitory activity of RGPF, FPK, or NPW active peptides was evaluated using a cyclooxygenase-2 inhibitor screening kit (Beyotime, Shanghai, China).

[0037] 75 μL COX2AssayBuffer, 5 μL COX2Cofactor working solution, 5 μL recombinant human COX2 enzyme solution, and 5 μL peptide sample solutions of different concentrations (100, 250, 500, 1000, 2000, and 4000 μM) were sequentially added to a 96-well plate. The peptide sample solutions were diluted with pure water to different concentrations, mixed well, and pre-incubated at 37°C for 10 min. Then, 5 μL COX2Probe and 5 μL COX2Substrate working solutions were added, and the reaction was carried out at 37°C for 5 min. The relative fluorescence units (RFU) of the reaction system were measured using a microplate reader at an excitation wavelength of 560 nm and an emission wavelength of 590 nm. In the 100% enzyme activity control group, COX2AssayBuffer was used instead of the recombinant human COX2 enzyme solution, and in the blank control group, pure water was used instead of the peptide sample solution. The amino acid sequences of the peptide samples were RGPF, FPK, or NPW, respectively.

[0038] COX2 inhibition rate (%) = (RFU) 100％酶活性对照 -RFU 样品 )÷(RFU 100％酶活性对照 -RFU 空白对照 )×100%.

[0039] IC 50 The value represents the sample concentration at which COX2 activity is inhibited by 50%.

[0040] The COX2 inhibition rates of active peptides RGPF, FPK, and NPW are shown in Table 1.

[0041] Table 1. Inhibition rate of different bioactive peptides on COX2 at different concentrations

[0042] Concentration (μM) RGPF FPK NPW 100 19.51±1.39 10.42±3.64 10.95±3.08 250 40.13±3.44 32.13±2.33 19.25±2.40 500 60.28±2.90 47.39±0.78 38.81±2.20 1000 74.83±1.82 60.37±1.41 50.11±3.35 2000 85.76±2.79 70.88±1.57 59.36±0.33 4000 91.41±3.88 83.65±1.98 73.88±2.10

[0043] The results in Table 1 show that the active peptides with amino acid sequences of RGPF, FPK, or NPW can effectively inhibit COX2.

[0044] Figure 1 The results showed that the IC50 values ​​of the active peptides RGPF, FPK, and NPW for inhibiting COX2 activity were [missing information]. 50 The values ​​were 360.17±32.60μM, 640.33±10.29μM, and 1100.90±89.93μM, respectively.

[0045] In summary, the bioactive peptides RGPF, FPK, and NPW all possess the ability to effectively inhibit COX2 activity, making them COX2 inhibitory peptides. By effectively inhibiting COX2 activity, these peptides can be used to prevent and / or treat COX2-induced inflammation or cancer, while also having antipyretic and analgesic effects.

[0046] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. The use of a COX2 inhibiting peptide or a composition inhibiting COX2 for the preparation of a product for the prevention and / or treatment of a COX2-induced disease, characterized in that, The disease is inflammation; The amino acid sequence of the COX2 inhibiting peptide is RGPF, FPK or NPW; The active ingredients of the composition include two or three of RGPF, FPK and NPW.

2. Use of a COX2 inhibiting peptide or a composition inhibiting COX2 for the preparation of a product for the prevention and / or treatment of pain and / or fever caused by COX2, characterized in that, The amino acid sequence of the COX2 inhibiting peptide is RGPF, FPK or NPW; The active ingredients of the composition include two or three of RGPF, FPK and NPW.

3. Use according to claim 1 or 2, characterized in that, The composition further includes pharmaceutically acceptable excipients.

4. Use according to claim 1 or 2, characterized in that, The only active ingredients of the composition are one or more of the RGPF, FPK and NPW.

5. Use according to claim 1 or 2, characterized in that, The product includes a reagent or a drug.

6. Use according to claim 1 or 2, characterized in that, The dosage form of the product includes any pharmaceutically acceptable dosage form.

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