New application of PDRN, composition of PDRN and application of PDRN

Abstract

The invention discloses a novel application of PDRN, a composition of PDRN and application of PDRN, and relates to the technical field of biology, and the composition comprises PDRN and beta-alanyl hydroxyprolyl diaminobutyric acid benzylamine. The PDRN or the composition can inhibit the activity of angiotensin converting enzyme and improve or eliminate edema type under-eye puffiness.

Description

TECHNICAL FIELD

[0001] The present disclosure relates to the field of biotechnology, in particular to a new use of PDRN and compositions and applications thereof. BACKGROUND

[0002] Polydeoxyribonucleotide (PDRN) refers to a deoxyribonucleic acid (DNA) fragment with biological activity in a broad sense. In the fields of skincare, biomedicine, etc., PDRN usually refers to a specific DNA fragment with a molecular weight range of about 50-1500 kDa. The base composition of PDRN is highly homologous to human DNA, and thus PDRN exhibits significant biological activity in human body. Currently, commercially available PDRN is mainly derived from the gonads or sperm cells of specific salmonids (e.g., rainbow trout Oncorhynchus mykiss or Chinook salmon Oncorhynchus keta). In addition to animal-derived PDRN, plant-derived PDRN has also attracted increasing attention from researchers. For example, Korean patent KR102682941B1 discloses a PDRN produced by rose plant cells. Recently, two other Korean studies reported PDRN derived from Chlorella protothecoides (JM Park, GB Nam, ES Lee, et al. Effects of Chlorella protothecoides-derived polydeoxyribonucleotides on skin regeneration and wound healing [J]. Archives of Dermatological Research, 2025) and PDRN derived from Gynostemma pentaphyllum callus (E Kim, S Choi, J Jang, et al. Effects of polydeoxyribonucleotide derived from Gynostemma pentaphyllum callus on skin barrier function [J]. Biochemical and Biophysical Research Communications, 2025). The results of the studies show that plant-derived PDRN can achieve similar biological activity to animal-derived PDRN at the cellular level through the adenosine A2a receptor pathway.

[0003] Previous studies have shown that PDRN has a variety of positive biological activities, one of the core mechanisms of which involves acting as an adenosine A2a receptor agonist, activating downstream signaling pathways (such as the PI3K / Akt pathway), thereby exerting a significant anti-inflammatory effect. By activating the A2a receptor, PDRN can also effectively inhibit the activity of High Mobility Group Box 1 (HMGB-1). HMGB-1 is an important pro-inflammatory and pro-fibrotic factor, and its overexpression is closely related to chronic inflammation and pathological scarring. By inhibiting HMGB-1, PDRN not only enhances its anti-inflammatory effect, but also significantly promotes the ordered deposition and remodeling of collagen, thereby effectively preventing and reducing scar hyperplasia. In addition, PDRN can effectively promote tissue repair and regeneration, and its role mainly manifests in promoting the proliferation of fibroblasts and promoting the synthesis and secretion of key components of the extracellular matrix (ECM) such as type I collagen, type III collagen, fibronectin, and elastin. These characteristics enable PDRN to exhibit a positive effect in promoting wound healing.

[0004] It is known that PDRN has significant efficacy in anti-inflammatory, prevention of scar hyperplasia, and promotion of tissue repair. However, given the unique biological activity and mechanism of action of PDRN, exploring its new uses beyond the known efficacy and providing more possibilities for efficacy are still important technical problems to be further studied and solved in the art. SUMMARY

[0005] The present disclosure relates to new uses of PDRN and compositions and applications thereof, in particular to a composition comprising PDRN and β-alanylhydroxyprolyl diaminobutyric acid benzylamine, which can inhibit angiotensin-converting enzyme (ACE) activity and improve or eliminate edema-type eye bags.

[0006] To help those skilled in the art correctly understand the term "edema-type eye bag" in the present disclosure, the term is explained in detail as follows.

[0007] The cause of edema type eye bags is related to local microcirculation disorder around the eyes and poor lymphatic return, leading to accumulation of body fluid. Lymphatic drainage is affected by bradykinin, a circulating nonapeptide that stimulates lymphatic drainage by activating the muscles surrounding the vascular network. The level of bradykinin in circulation is regulated by angiotensin-converting enzyme, which rapidly degrades bradykinin, thereby causing drainage disorders and inducing edema. On the other hand, angiotensin-converting enzyme (ACE) is a key enzyme in the renin-angiotensin system (RAS) that can hydrolyze angiotensin I to angiotensin II. Angiotensin II has a strong vasoconstrictor effect, and at the same time increases the sensation of thirst and water retention in the vascular system. Therefore, by inhibiting the activity of angiotensin-converting enzyme, the level of angiotensin II can be reduced, the microcirculation of the skin around the eyes can be improved, and at the same time the level of bradykinin can be increased, improving lymphatic circulation, thereby promoting the excretion of harmful fluids in the skin, eliminating edema, and improving or eliminating edema type eye bags.

[0008] In one aspect, the present disclosure provides a composition comprising PDRN and beta-alanyl hydroxyprolyl diaminobutyroyl benzylamide.

[0009] Beta-alanyl hydroxyprolyl diaminobutyroyl benzylamide (INCI name: Beta-Alanyl Hydroxyprolyl Diaminobutyroyl Benzylamide, Guoqing Original Preparation No. 20210003) is a polypeptide derivative that can increase skin elasticity, which is a product independently developed by Shenzhen Weiqi Technology Co., Ltd. The present inventors have surprisingly found that PDRN has a very strong inhibitory effect on ACE activity in addition to its anti-inflammatory repair effect, and can play a role in improving or eliminating edema type eye bags; in particular, when PDRN and beta-alanyl hydroxyprolyl diaminobutyroyl benzylamide are combined, the two produce a synergistic effect, further enhancing the ability of PDRN to inhibit ACE activity, which is conducive to enhancing the effect of improving or eliminating edema type eye bags.

[0010] In some embodiments, the concentration ratio of PDRN and beta-alanyl hydroxyprolyl diaminobutyroyl benzylamide is (3-4): 1.

[0011] In some embodiments, the concentration ratio of PDRN and beta-alanyl hydroxyprolyl diaminobutyroyl benzylamide is 4: 1.

[0012] In some embodiments, the PDRN is of animal origin.

[0013] In some embodiments, the PDRN is of rainbow trout, Chinook salmon and / or Atlantic salmon origin.

[0014] The base sequence of PDRN of animal origin is not exactly the same as that of PDRN of plant origin, and the PDRN of animal origin has up to 98% similarity with the DNA base sequence of human. After experimental verification, it is found that the PDRN of animal origin, especially the PDRN derived from Oncorhynchus mykiss, Oncorhynchus keta or Salmo salar, has a significant ability to inhibit ACE activity. After the PDRN of animal origin is combined with β-alanyl hydroxyproline diamino butyric acid benzylamine, the combination produces a synergistic effect and has a better ability to inhibit ACE activity. However, the PDRN of plant origin, especially the PDRN derived from rose, when combined with β-alanyl hydroxyproline diamino butyric acid benzylamine, exhibits an ability to enhance ACE activity. Apparently, the effects of combinations of PDRN of different origins with β-alanyl hydroxyproline diamino butyric acid benzylamine on ACE activity are quite different.

[0015] In another aspect of the present disclosure, a cosmetic product is provided, which comprises an effective amount of the above-mentioned composition and at least one additional ingredient.

[0016] "Effective amount" means an amount of the composition of the present disclosure that is non-toxic but sufficient to provide the desired effect. The composition of the present disclosure is used in the cosmetic product at an effective concentration to achieve the desired effect. In some embodiments, the concentration of the composition is between 0.00000001% (by weight) and 20% (by weight) relative to the total weight of the cosmetic product; in some embodiments, the concentration of the composition is between 0.000001% (by weight) and 15% (by weight) relative to the total weight of the cosmetic product; in some embodiments, the concentration of the composition is between 0.0001% (by weight) and 10% (by weight) relative to the total weight of the cosmetic product; in some embodiments, the concentration of the composition is between 0.0001% (by weight) and 5% (by weight) relative to the total weight of the cosmetic product.

[0017] In some embodiments, the dosage form of the cosmetic product includes a paste, a cream, an emulsion, an aqueous agent, an oil agent, a gel, a powder, a tablet, a mud, a patch, a film, an aerosol, a spray, a lyophilized preparation or a nano-preparation.

[0018] In another aspect of the present disclosure, the use of PDRN or the above-mentioned composition in the preparation of a composition for inhibiting angiotensin converting enzyme activity is provided.

[0019] In another aspect of the present disclosure, the use of PDRN or the above-mentioned composition in the preparation of a composition for improving or eliminating edema-type eye bags is provided.

[0020] In some embodiments, the PDRN is an animal-derived PDRN.

[0021] In some embodiments, the PDRN is an animal-derived PDRN.

[0022] In another aspect of the present disclosure, there is provided a use of the above-mentioned composition in the preparation of a cosmetic product.

[0023] In another aspect of the present disclosure, there is provided a cosmetic method for non-therapeutic purposes, the method comprising using the above-mentioned composition, or the above-mentioned cosmetic product, on the skin.

[0024] The present disclosure has the following advantages and effects:

[0025] 1. It has been verified through experiments that PDRN, especially animal-derived PDRN, can effectively inhibit ACE activity and can be used to improve or eliminate edema-type eye bags.

[0026] 2. The combination of PDRN and β-alanyl hydroxyproline diamino butyric acid benzylamine of the present disclosure produces a synergistic effect and has a better technical effect of inhibiting ACE activity, and can be used to improve or eliminate edema-type eye bags. DETAILED DESCRIPTION

[0027] In order to make the objectives, characteristics and advantages of the present disclosure more obvious and easy to understand, the present disclosure will be further described in detail below in combination with embodiments. Obviously, the described embodiments are part of the embodiments of the present disclosure, rather than all the embodiments. Based on the embodiments in the present disclosure, all other embodiments obtained by a person of ordinary skill in the art without making creative efforts fall within the scope of the claims of the present disclosure.

[0028] The salmon PDRN used in the embodiments of the present disclosure is derived from Oncorhynchus keta and has a molecular weight of 60-100 kDa, which can be purchased from Nanjing Letuo Biological Technology Co., Ltd. The rose PDRN used in the embodiments of the present disclosure can be purchased from Guangzhou Xinyingke Chemical Co., Ltd. The experimental reagents and materials used in the present disclosure can be obtained through commercial channels.

[0029] The compounding ratio of each component in the composition of the embodiments of the present disclosure is shown in Table 1:

[0030] Table 1

[0031]

[0032]

[0033] Note: When the concentration of the composition is 100 ppm, such as salmon PDRN: β-alanyl hydroxyprolyl diaminobutyric acid benzylamine = 3:1, it means that the concentration of salmon PDRN is 75 ppm and the concentration of β-alanyl hydroxyprolyl diaminobutyric acid benzylamine is 25 ppm.

[0034] Example 1 ACE activity inhibition test

[0035] 1.1 Reagents and materials

[0036] PBS buffer, N-[3-(2-furyl)acryloyl]-L-phenylalanyl glycyl glycine (FAPGG), angiotensin converting enzyme (ACE).

[0037] 1.2 Instruments

[0038] Microplate reader, air bath constant temperature shaker.

[0039] 1.3 Test samples and grouping

[0040] 1.3.1 Test samples

[0041] Composition 1, composition 2, composition 3, composition 4, salmon PDRN, β-alanyl hydroxyprolyl diaminobutyric acid benzylamine, and all the above test samples have a test concentration of 100 ppm.

[0042] 1.3.2 Grouping

[0043] Sample group: test sample, FAPGG, ACE;

[0044] Blank control group: PBS, FAPGG, ACE.

[0045] 1.4 Experimental method

[0046] ACE was prepared with secondary purified water, and FAPGG was dissolved with PBS buffer with a pH value of 8.3.

[0047] The test sample was diluted to the required concentration with PBS buffer with a pH value of 8.3. A 96-well plate was taken, 50 μL of the test sample and 50 μL of the FAPGG solution (5 mmol / L) were added to the sample group, and 50 μL of the PBS buffer (pH = 8.3) and 50 μL of the FAPGG solution (5 mmol / L) were added to the blank control group. The absorbance value S1 of the sample group and the absorbance value C1 of the blank control group were measured at 340 nm. The sample group and the blank control group were incubated at 37°C for 30 min, then ACE (0.1 U / mL) was added, and the incubation was continued at 37°C for 30 min. Then the absorbance values S2 and C2 of the sample group and the blank control group were measured at 340 nm. The ACE activity inhibition rate was calculated according to the following formula:

[0048]

[0049] 1.5 Experimental results

[0050] FAPGG is an enzymatic reaction substrate of ACE, and under the catalysis of ACE, FAPGG will undergo hydrolysis to cause its color to gradually disappear, thereby showing a result of a decrease in absorbance value. In this experiment, ACE was treated with the sample to be tested, and the reaction amount of FAPGG was determined by detecting the change in the absorbance value of FAPGG, so as to determine whether the sample to be tested of the present disclosure can inhibit the activity of ACE. The ACE activity inhibition rate of the test sample is shown in Table 2.

[0051] Table 2 ACE activity inhibition rate

[0052]

[0053]

[0054] The results show that the inhibition rate of salmon PDRN on ACE activity is as high as 83.56% at 100 ppm, proving that PDRN of animal origin has a significant effect of inhibiting ACE activity and can be used to improve or eliminate edema-type eye bags.

[0055] In addition, the results in Table 2 also show that β-alanyl hydroxyprolyl diaminobutyric acid benzylamine does not show a significant effect on ACE activity, however, compositions 2 and 3 obtained by combining salmon PDRN and β-alanyl hydroxyprolyl diaminobutyric acid benzylamine both show a significantly better inhibitory effect on ACE activity than salmon PDRN alone, proving that the combination of compositions 2 and 3 produces a significant synergistic effect. The inhibition rate of composition 1 on ACE activity is not significantly different from the inhibition rate of salmon PDRN alone on ACE activity. Composition 4 shows not only no inhibitory effect on ACE activity but also a promoting effect on ACE activity after combining rose PDRN and β-alanyl hydroxyprolyl diaminobutyric acid benzylamine, proving that the effect produced by the combination of PDRN of different origins and β-alanyl hydroxyprolyl diaminobutyric acid benzylamine is unpredictable. The animal-derived PDRN of the present disclosure and β-alanyl hydroxyprolyl diaminobutyric acid benzylamine produce an unexpected effect of inhibiting ACE activity when combined, and when they are combined at a concentration ratio of (3-4): 1, a significant synergistic effect is produced, which can be used to improve or eliminate edema-type eye bags.

[0056] In this disclosure, relational terms such as first and second and the like can be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises", "comprising", or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but can include other elements not expressly listed or inherent to such process, method, article, or apparatus. An element proceeded by "comprises... a" does not, without more constraints, exclude the existence of additional identical elements in the process, method, article, or apparatus that comprises the element.

[0057] While particular embodiments of the present disclosure have been described, alternatives, modifications, and improvements can be made to those embodiments without departing from the spirit and scope of the present disclosure. Such alternatives, modifications, and improvements have been considered to fall within the scope of the present disclosure.

Claims

1. Composition, characterized in that, PDRN and beta-alanyl hydroxyprolyl diaminobutyric acid benzylamide.

2. The composition of claim 1, wherein, The concentration ratio of the PDRN and beta-alanyl hydroxyprolyl diaminobutyric acid benzylamide is (3-4):

1.

3. The composition of claim 1, wherein, The concentration ratio of the PDRN and beta-alanyl hydroxyprolyl diaminobutyric acid benzylamide is 4:

1.

4. The composition according to any one of claims 1 to 3, characterized in that, The PDRN is of animal origin.

5. The composition according to any one of claims 1 to 3, characterized in that, The PDRN is of rainbow trout, chinook salmon and / or Atlantic salmon origin.

6. A cosmetic product, characterized by, The cosmetic comprises an effective amount of the composition of any one of claims 1-5, and at least one additional ingredient.

7. The cosmetic product according to claim 6, characterized in that, The dosage form of the cosmetic comprises a cream, a lotion, an emulsion, an aqueous agent, an oil agent, a gel, a powder, a tablet, a mud, a patch, a film, an aerosol, a spray, a lyophilized preparation or a nano-preparation.

8. Use of PDRN or the composition of any one of claims 1-5 in the preparation of a composition for inhibiting angiotensin converting enzyme activity.

9. Use of PDRN or the composition of any one of claims 1-5 in the preparation of a composition for improving or eliminating edema type eye bags.

10. Use according to claim 8 or 9, characterized in that, The PDRN is of animal origin.

11. Use according to claim 8 or 9, characterized in that, The PDRN is of rainbow trout, chinook salmon and / or Atlantic salmon origin.

12. Use of the composition of any one of claims 1-5 in the preparation of a cosmetic.

13. A cosmetic method for non-therapeutic purposes, characterized in that, The method comprises using the composition of any one of claims 1-5, or the cosmetic of claim 6 or 7 on the skin.

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