COX2 inhibitory peptide, composition for inhibiting COX2 and application

Abstract

The invention provides a COX2 inhibitory peptide, a composition for inhibiting COX2 and application, and belongs to the technical field of bioactive peptides. The amino acid sequence of the COX2 inhibitory peptide comprises RGPF, FPK or NPW. Research finds that IC50 values of COX2 activity inhibition of the COX2 inhibitory peptide with the amino acid sequence shown as RGPF, FPK or NPW are 360.17 + / -32.60 [mu] M, 640.33 + / -10.29 [mu] M and 1100.90 + / -89.93 [mu] M respectively, and the COX2 inhibitory peptide can effectively inhibit the activity of COX2, so that inflammation and cancers caused by COX2 are effectively prevented and / or treated, and the purposes of antipyretic and analgesic can be achieved. Meanwhile, the COX2 inhibitory peptide disclosed by the invention has the advantages of safety, no toxicity, good water solubility and the like, and provides a choice for safely using a COX2 inhibitor clinically.

Description

Technical Field

[0001] The present invention belongs to the technical field of bioactive peptides, and in particular relates to a COX2 inhibitory peptide, a COX2 inhibiting composition and an application thereof. Background Art

[0002] Cyclooxygenase-2 (COX2) is an inducible cyclooxygenase, part of the COX enzyme class, and participates in the synthesis of prostaglandins together with COX1. COX2 is expressed in a variety of tissues, but is usually expressed at a low level in non-inflammatory areas. Growth factors, cytokines, bacterial lipopolysaccharides, etc. can stimulate a significant increase in the expression of COX2, catalyzing the conversion of arachidonic acid into prostaglandins, which in turn causes a series of adverse reactions such as inflammation, pain, and even cancer.

[0003] Studies have found that COX2 expression and enzyme activity are significantly increased 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. The use of COX2 inhibitors can effectively alleviate the symptoms of related diseases and affect the progression of the disease to a certain extent.

[0004] At present, COX2 inhibitors are mainly chemically synthesized non-steroidal anti-inflammatory drugs, including salicylates, aromatic or heterocyclic aromatics, hydroxybenzothiazines (coxibs), etc. However, these drugs, while having COX2 inhibitory activity, also increase the possibility of thrombotic cardiovascular disease. In 2005, the US FDA believed that the risk of cardiovascular events increased by coxibs was unavoidable and recommended the suspension of the sale of valdecoxib.

[0005] As the research deepens, some natural compounds have also 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 exert 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 have multiple biological activities and high nutritional value. More importantly, peptides are non-toxic, safer, and easier to digest and absorb. However, there are no reports on the use of COX2 inhibitory peptides in the preparation of COX2 inhibitory drugs or reagents. Summary of the invention

[0007] In view of this, the object of the present invention is to provide a COX2 inhibitory peptide, a COX2 inhibiting composition and use, wherein the COX2 inhibitory peptide has a COX2 inhibitory effect.

[0008] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

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

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

[0011] Preferably, the composition further comprises a pharmaceutically acceptable excipient.

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

[0013] The present invention also provides a use of the COX2 inhibitory peptide or composition in preparing a product for inhibiting COX2.

[0014] The present invention also provides a use of the above COX2 inhibitory peptide or composition in the preparation of products for preventing and / or treating diseases caused by COX2.

[0015] The present invention also provides a use of the above COX2 inhibitory peptide or composition in the preparation of a product for preventing and / or treating pain and / or fever caused by COX2.

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

[0017] Preferably, the product comprises a reagent or a medicine.

[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] The present invention provides a COX2 inhibitory peptide, a composition for inhibiting COX2 and its application. The present invention finds through research that the IC value of the COX2 inhibitory peptide represented by the amino acid sequence such as RGPF, FPK or NPW for inhibiting COX2 activity is 50 The values ​​are 360.17±32.60μM, 640.33±10.29μM and 1100.90±89.93μM, respectively, which can effectively inhibit the activity of COX2, thereby effectively preventing and / or treating inflammation and cancer caused by COX2, and can also achieve the purpose of antipyretic and analgesic. At the same time, the COX2 inhibitory peptide of the present invention has the advantages of safety, non-toxicity, good water solubility, etc., and provides a new choice for the safe use of COX2 inhibitors in clinical practice. BRIEF DESCRIPTION OF THE DRAWINGS

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

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

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

[0024] In the present invention, the composition also includes pharmaceutically acceptable excipients. The excipients include excipients. The excipients broadly refer to any component other than the effective active ingredient, and the excipients may be inert substances, inactive substances and / or substances without medical activity. Excipients may meet different purposes, such as as carriers, solvents, diluents, tablet adjuvants and / or improve the release and absorption of active ingredients, including one or more of solvents, diluents, buffers, preservatives, isotonic agents, pH regulators, chelating agents and stabilizers. The buffer includes one or more of Krebs buffer, sodium dihydrogen phosphate, sodium hydrogen phosphate, succinate, maleic acid, fumaric acid and tartaric acid. The preservative includes one or more of phenol, o-cresol, m-cresol, p-cresol, methyl parahydroxybenzoate, propyl parahydroxybenzoate, 2-phenoxyethanol, butyl parahydroxybenzoate, benzyl alcohol, benzoic acid, chlorocresol and chlorphenesin. The chelating agent may be selected from ethylenediaminetetraacetic acid or citric acid. The isotonic agent may be selected from one or more of salt (eg, sodium chloride), sugar alcohol (eg, glycerol, propylene glycol, 1,3-propanediol, 1,3-butanediol), and polyethylene glycol (eg, PEG400).

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

[0026] The present invention also provides a use of the COX2 inhibitory peptide or composition in preparing a product for inhibiting COX2.

[0027] The present invention also provides a use of the above COX2 inhibitory peptide or composition in the preparation of products for preventing and / or treating diseases caused by COX2.

[0028] The present invention also provides a use of the above COX2 inhibitory peptide or composition in the preparation of a product for preventing and / or treating pain and / or fever caused by COX2.

[0029] As an implementation method, the disease includes one or both of inflammation and cancer. The product includes an agent or a drug. The product can be used to inhibit the activity of COX2 using a COX2 inhibitory peptide or composition as the only active ingredient, or can be used in combination with other ingredients that inhibit the activity of COX2 to inhibit the activity of COX2, thereby achieving the effect of preventing and / or treating inflammation and cancer, and also having antipyretic and analgesic effects. The present invention has found that RGPF, FPK or NPW active peptides can effectively inhibit the activity of COX2, and the IC of COX2 activity inhibition is 50 The values ​​were 360.17±32.60 μM, 640.33±10.29 μM and 1100.90±89.93 μM, respectively.

[0030] In the present invention, the dosage form of the product includes any pharmaceutically acceptable dosage form, such as a sustained-release agent, a capsule, a granule, an oral solution, a tablet or an injection. The product can be administered orally or by injection. The mass ratio of the COX2 inhibitory peptide or composition to the product is 50% to 100%, such as 50%, 60%, 70%, 80%, 90% or 100%. The product also includes pharmaceutically acceptable excipients, and the types of the excipients refer to the types of pharmaceutically acceptable excipients mentioned in the above-mentioned composition, which will not be repeated here.

[0031] In the present 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 are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0033] Example 1

[0034] Activity assay of COX2 inhibitory peptides

[0035] In this example, the amino acid sequence of the COX2 inhibitory peptide is RGPF (SEQ ID NO. 1), FPK or NPW, which are 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 COX-2 inhibitor screening kit (Beyotime, Shanghai, China).

[0037] 75 μL COX2AssayBuffer, 5 μL COX2Cofactor working solution, 5 μL human recombinant COX2 enzyme solution and 5 μL peptide sample solutions of different concentrations (100, 250, 500, 1000, 2000, 4000 μM) were added to a 96-well plate in sequence. The peptide sample solutions were diluted with pure water to different concentrations, mixed and pre-incubated at 37°C for 10 min. 5 μL COX2Probe and 5 μL COX2Substrate working solutions were added and reacted at 37°C for 5 min. The relative fluorescence unit (RFU) value of the reaction system was determined by a microplate reader, with an excitation wavelength of 560 nm and an emission wavelength of 590 nm. In the 100% enzyme activity control group, COX2AssayBuffer was used to replace the human recombinant COX2 enzyme solution, and pure water was used to replace the peptide sample solution in the blank control group. Among them, 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 is the sample concentration at which COX2 activity is inhibited by 50%.

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

[0041] Table 1 Inhibition rate of COX2 by different active peptides 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 indicate that the active peptides with the amino acid sequences of RGPF, FPK or NPW can effectively inhibit COX2.

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

[0045] In summary, the active peptides RGPF, FPK and NPW all have the ability to effectively inhibit COX2 activity and are COX2 inhibitory peptides. The active peptides RGPF, FPK and NPW can effectively inhibit COX2 activity to prevent and / or treat inflammation or cancer caused by COX2, and also have antipyretic and analgesic effects.

[0046] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A COX2 inhibitory peptide, It is characterized in that The amino acid sequence of the COX2 inhibitory peptide includes RGPF, FPK or NPW.

2. A composition for inhibiting COX2, It is characterized in that The active ingredients of the composition include one or more of the RGPF, FPK and NPW described in claim 1.

3. The composition according to claim 2, It is characterized in that The composition further includes a pharmaceutically acceptable excipient.

4. The composition according to claim 2, It is characterized in that The only active ingredient of the composition is one or more of RGPF, FPK and NPW as described in claim 1.

5. Use of the COX2 inhibitory peptide according to claim 1 or the composition according to any one of claims 2 to 4 in the preparation of a product that inhibits COX2.

6. Use of the COX2 inhibitory peptide according to claim 1 or the composition according to any one of claims 2 to 4 in the preparation of a product for preventing and / or treating diseases caused by COX2.

7. Use of the COX2 inhibitory peptide according to claim 1 or the composition according to any one of claims 2 to 4 in the preparation of a product for preventing and / or treating pain and / or fever caused by COX2.

8. The use according to claim 6, It is characterized in that The disease includes one or both of inflammation and cancer.

9. The use according to any one of claims 5 to 7, It is characterized in that The product includes a reagent or a drug.

10. The use according to any one of claims 5 to 8, It is characterized in that The dosage form of the product includes any pharmaceutically acceptable dosage form.

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