Cyclic heptapeptide as well as preparation method and application thereof

Abstract

The invention provides a cyclic heptapeptide, the amino acid sequence of the cyclic heptapeptide is cyclic Gly-Tyr-Tyr-Glu-Leu-Asn-Asp, the C terminal is amidated (-NH2), the molecular formula is C39H50N8O14, chain peptide resin is obtained through solid-phase synthesis of a peptide chain, a linear chain peptide crude product is obtained after splitting decomposition, a linear chain peptide is obtained after purification, a cyclic peptide crude product is obtained after cyclization, and / or the cyclic heptapeptide is prepared through finished product purification. The cyclic heptapeptide has an inflammation regulation effect, an anti-oxidation regulation effect and / or a telomere protection effect, forms an anti-inflammation-anti-oxidation-telomere protection synergistic anti-aging network, has high biological activity, a multi-target synergistic effect, high receptor binding efficiency and high stability, and is used for preparing cyclic peptide skincare products in the anti-aging field.

Description

Technical Field

[0001] This invention relates to the field of peptide compounds, specifically to a cyclic heptapeptide, its preparation method, and its applications. Background Technology

[0002] With the accelerating aging of the population, the market demand for anti-aging products is growing rapidly. Among the many active anti-aging ingredients, peptides have become a research hotspot in the field of anti-aging due to their specific biological activities and relatively high safety. However, existing peptide-based anti-aging products still have many shortcomings. For example, existing peptide products suffer from low receptor binding efficiency, limiting their biological effects. Furthermore, many peptide products exhibit poor stability in vivo, are easily degraded by proteases, resulting in short-lasting activity. Moreover, existing peptide products often target only a single aging mechanism, lacking multi-target synergistic effects, which falls far short of meeting practical needs.

[0003] Linear peptide products are widely used, but most lack sufficient bioactivity to effectively exert anti-aging effects. For example, heptapeptide-6, as a linear peptide, has advantages such as high activity and easy compatibility, but also has a series of weaknesses that require attention. These potential disadvantages are mostly closely related to its linear molecular structure. Firstly, heptapeptide-6 faces stability challenges: compared to cyclic peptides, linear peptides' peptide bonds are more easily recognized and degraded by proteases on the skin surface (such as various enzymes secreted by keratinocytes). This may lead to a lower actual effective concentration on the skin surface and a shorter duration of activity, thus affecting the lasting efficacy of the final product. Secondly, heptapeptide-6 is controversial regarding its transdermal absorption efficiency: although suppliers often claim that heptapeptide-6 has a high transdermal rate due to its small molecular weight, a small molecular weight alone does not completely equate to efficient transdermal absorption; it is also related to the lipid-soluble nature of its structure. Thirdly, heptapeptide-6 has the problem of single-target action: heptapeptide-6 mainly exerts its effects by activating specific signaling pathways (such as the TGF-β / Smad pathway). This relatively singular mechanism of action may mean that its anti-aging effects focus primarily on stimulating collagen regeneration, while its performance in improving dynamic wrinkles (such as through a botulinum toxin-like mechanism) or providing highly effective hydration may not be as strong as other specially designed peptides or ingredients. Furthermore, heptapeptide-6 formulations are highly dependent on the overall formula: its efficacy is heavily reliant on the support of the entire formulation. For example, whether to add effective penetration enhancers (such as lecithin or amino acid derivatives) to help it penetrate the skin barrier, and how to ensure its chemical stability and bioactivity within the formulation, all place high demands on the formulation process. Additionally, heptapeptide-6 carries a potential risk of irritation.

[0004] Cyclic peptides are polypeptides with a cyclic structure formed by covalent bonds between the N-terminus and C-terminus or side chains of the peptide chain. Compared with linear peptides, cyclic peptides have a more stable conformation, stronger resistance to enzymatic degradation, and higher bioavailability. Currently, cyclic peptide compounds are widely used in pharmaceuticals, cosmetics, and other fields. Due to their unique cyclic structure, cyclic peptide compounds also have significant advantages in terms of biological stability and targeting. However, there are still significant technical barriers to the screening of cyclic peptide compounds, the industrial application of synthetic methods, and product efficacy. For example, existing cyclic peptide synthesis methods are complex, difficult to purify, and have low yields, which are not conducive to industrial production.

[0005] Currently, there is an urgent need to develop an anti-aging product that overcomes the aforementioned linear peptide defects, possesses high activity, can be industrially produced, and has multiple effects, and to explore and establish an efficient and simple synthesis method to meet the growing market demand for anti-aging products. Summary of the Invention

[0006] To address the problems of single target, low receptor binding efficiency, and poor stability in existing linear peptide anti-aging products, such as heptapeptide-6, this invention provides a cyclic heptapeptide, its preparation method, and its application. This cyclic heptapeptide exhibits high bioactivity, multi-target synergistic effects, high receptor binding efficiency, and high stability, and can be used to prepare cyclic peptide-based beauty and skincare products in the field of anti-aging.

[0007] This invention provides a cyclic heptapeptide, wherein the amino acid sequence of the cyclic heptapeptide is cyclic Gly-Tyr-Tyr-Glu-Leu-Asn-Asp, the C-terminus is amidated (-NH2), and the molecular formula is C 39 H 50 N8O 14 .

[0008] Preferably, the present invention provides a cyclic heptapeptide, wherein the cyclic heptapeptide is prepared by solid-phase synthesis of peptide chains to obtain chain peptide resin, cleavage to obtain crude linear chain peptide, purification to obtain linear chain peptide, cyclization to obtain crude cyclic peptide, and / or purification of the finished product.

[0009] Preferably, the present invention provides a cyclic heptapeptide, wherein solid-phase synthesis includes swelling 2-chlorotriphenylmethyl chloride (CTC) resin with dichloromethane (DCM), and sequentially coupling amino acids starting from Asp to form a chain peptide resin; cleavage includes cleaving the chain peptide resin with 10%~40% hexafluoroisopropanol (HFIP) / dichloromethane (DCM) to obtain a linear chain peptide crude product; purification includes purifying the crude product by reversed-phase high-performance liquid chromatography (HPLC) to obtain a linear chain peptide; Cyclization involves dissolving the linear peptide in an organic solvent, including N,N-dimethylformamide (DMF) or dichloromethane (DCM), adding 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) and / or N,N-diisopropylethylamine (DIEA) for cyclization, monitoring the reaction, and obtaining a crude cyclized peptide. Purification of the final product involves cleaving the crude cyclized peptide for byproducts, washing, high-performance liquid chromatography (HPLC) purification, and / or lyophilization to obtain a cyclic heptapeptide.

[0010] Preferably, the present invention provides a cyclic heptapeptide, wherein the coupled amino acids include linking Asp to a resin, followed by deprotection and activation with O-benzotriazole-tetramethylurea hexafluorophosphate (HBTU); the deprotection uses a solution including piperidine (PIP) and its derivatives, N,N-dimethylformamide (DMF) and / or trifluoroacetic acid (TFA) as a deprotecting agent; the cyclization process is monitored using liquid chromatography-mass spectrometry (LCMS); the byproducts are cleaved during the purification of the finished product using a solution of trifluoroacetic acid (TFA) as a cleavage agent.

[0011] Preferably, the present invention provides a cyclic heptapeptide, wherein the cyclic heptapeptide has inflammatory regulation, antioxidant regulation, and / or telomere protection effects, forming a synergistic anti-aging network of "anti-inflammatory-antioxidant-telomere protection", which can be used to prepare beauty and skin care products in the field of anti-aging.

[0012] Preferably, the cyclic heptapeptide provided by the present invention has an inflammatory regulatory effect including binding to multiple sites of the pro-inflammatory cytokine high mobility group box 1 (HMGB1), the sites being located on a protein functional domain (A-box) with a specific three-dimensional spatial structure and consisting of approximately 79 amino acid residues; causing allosteric changes in high mobility group box 1 (HMGB1), inhibiting the release of high mobility group box 1 (HMGB1), and / or inhibiting the binding of high mobility group box 1 (HMGB1) to Toll-like receptor 4 (TLR4), and / or inhibiting the binding of high mobility group box 1 (HMGB1) to the receptor for advanced glycation end products (RAGE) target site, and / or blocking the activation of κ-light chain enhancement (NF-κB) in downstream nuclear factor-activated B cells.

[0013] Preferably, the present invention provides a cyclic heptapeptide that binds to multiple sites of the pro-inflammatory cytokine high mobility group box 1 (HMGB1), the sites including alanine at position 17, arginine at position 10, and / or glycine at position 11.

[0014] Preferably, the present invention provides a cyclic heptapeptide wherein antioxidant regulation includes competitive binding to Kelch-like ECH-associated protein 1 (Keap1) and / or activation of the expression of nuclear factor E2-associated factor 2 (Nrf2).

[0015] Preferably, the present invention provides a cyclic heptapeptide, wherein the telomere protection effect includes promoting the expression of telomere repeat binding factor 1 (TRF1).

[0016] Preferably, the present invention provides a cyclic heptapeptide, wherein the cyclic heptapeptide can be used to prepare cosmetic and / or beauty and care products, including essence water, toner, lotion, face cream, beauty cream, base makeup, concealer, primer, setting makeup, eyeshadow, contouring, blush, lipstick, single-use ampoule, freeze-dried powder, freeze-dried liquid, facial mask liquid, facial mask powder, frozen mask, and / or functional medical beauty products.

[0017] Therefore, the cyclic heptapeptide provided by this invention possesses high bioactivity, multi-target synergistic effects, high receptor binding efficiency, and high stability. Simultaneously, this invention provides a method for preparing cyclic heptapeptides that is simple to operate, has a stable process, and can efficiently produce high-purity cyclic heptapeptides that meet practical needs. This method solves the difficulties in current cyclic peptide production processes, such as low cyclization reaction efficiency, difficulty in controlling side reactions, strong sequence dependence, significant challenges in solubility and purification, and high barriers to large-scale production, and has significant economic value.

[0018] The cyclic heptapeptide of this invention exhibits significant anti-aging effects, including anti-inflammatory aging, telomere protection, and antioxidant activity. Compared to existing linear peptide anti-aging products, the cyclic molecular conformation of the cyclic heptapeptide of this invention endows it with significantly superior receptor binding efficiency and bioactivity compared to linear peptides.

[0019] Therefore, the cyclic heptapeptide of this invention achieves a synergistic anti-aging network of "anti-inflammatory-antioxidant-telomere protection," realizing multi-dimensional anti-aging through a triple synergistic pathway: effectively blocking the interaction between HMGB1 and inflammation receptors, reducing chronic inflammatory damage at its source; enhancing the cell's ability to resist oxidative stress and improving the level of endogenous antioxidant defense; effectively maintaining telomere structural stability and delaying the cellular aging process, and can be used to prepare cosmetic skincare products in the field of anti-aging. In terms of inflammation regulation, the cyclic heptapeptide can bind to multiple sites on the A-box domain of the HMGB1 protein, including alanine at position 17, arginine at position 10, and glycine at position 11, effectively blocking the interaction between HMGB1 and TLR4 / RAGE; in terms of antioxidant regulation, the cyclic heptapeptide can competitively bind to Keap1, activating the expression of Nrf2; in terms of telomere protection, the cyclic heptapeptide can promote the expression of TRF1 and maintain the stability of telomere structure. Experimental data show that the cyclic heptapeptide of the present invention has significant efficacy in inhibiting HMGB1 release, inhibiting TLR4 gene expression, inhibiting RAGE gene expression, promoting Nrf2 gene expression, and promoting TRF1 gene expression. Attached Figure Description

[0020] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the following description of the embodiments will be briefly introduced. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0021] Figure 1 The structural information of the cyclic heptapeptide prepared in Example 3; Figure 2 The data are from the liquid chromatography-mass spectrometry analysis of the cyclic heptapeptide prepared in Example 3; Figure 3 This provides three-dimensional information on the binding of cyclic heptapeptide to multiple sites on the A-box functional domain of HMGB1. Figure 4 The above are fluorescence microscopy images showing the contrast between cyclic heptapeptide and heptapeptide-6 in inhibiting HMGB1 release in Example 5. Figure 5 The results of the experiment in Example 5 on the inhibition of TLR4 gene expression by cyclic heptapeptide; Figure 6 The results of the experiment in Example 5 on the inhibition of RAGE gene expression by cyclic heptapeptide; Figure 7 The results of the experiment in Example 6 on the promotion of Nrf2 gene expression by cyclic heptapeptide; Figure 8 The results of the experiment in Example 6 show the effect of cyclic heptapeptide promoting TRF1 gene expression. Detailed Implementation

[0022] To further illustrate the present invention, embodiments are given below. It should be noted that these embodiments are entirely illustrative. The purpose of providing these embodiments is to fully demonstrate the meaning and content of the present invention, but they do not limit the present invention to the scope of these embodiments. The technical solutions in the embodiments of the present invention will now be clearly and completely described in conjunction with the accompanying drawings. Example 1

[0023] This embodiment provides a cyclic heptapeptide with the amino acid sequence of cyclic Gly-Tyr-Tyr-Glu-Leu-Asn-Asp, an amidation (-NH2) at the C-terminus, and a molecular formula of C1. 39 H 50 N8O 14 .

[0024] This cyclic heptapeptide was prepared through steps including solid-phase synthesis, cleavage, purification, cyclization, and final product purification. The specific preparation process is as follows: First, solid-phase synthesis was performed using dichloromethane (DCM) to swell 2-chlorotriphenylmethyl chloride (CTC) resin. Amino acids were sequentially coupled, starting with Asp, to form a chain-like peptide resin. During amino acid coupling, Asp was attached to the resin, followed by deprotection using O-benzotriazole-tetramethylurea hexafluorophosphate (HBTU). The deprotection process employed a solution of piperidine (PIP) and its derivatives, N,N-dimethylformamide (DMF), and trifluoroacetic acid (TFA).

[0025] The process then proceeds to a cleavage step, in which the chain peptide resin is cleaved using 20% ​​hexafluoroisopropanol (HFIP) / dichloromethane (DCM) to obtain crude linear peptide products. The amount of hexafluoroisopropanol (HFIP) can be adjusted within the range of 10% to 40%, preferably 20%.

[0026] Then, a purification step was performed, in which the crude product was purified by reversed-phase high-performance liquid chromatography (HPLC) to obtain a linear peptide.

[0027] The subsequent cyclization step involves dissolving the linear peptide in an organic solvent such as N,N-dimethylformamide (DMF) or dichloromethane (DCM), and then adding 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) and N,N-diisopropylethylamine (DIEA) for cyclization. The reaction is monitored by liquid chromatography-mass spectrometry (LCMS) to obtain the crude cyclized peptide.

[0028] Finally, the product was purified by cleaving the crude cyclic peptide with trifluoroacetic acid (TFA) solution to remove byproducts, washing, high-performance liquid chromatography purification, and freeze-drying to obtain the cyclic heptapeptide product.

[0029] This cyclic heptapeptide possesses multiple bioactivities, forming a synergistic anti-aging network of "anti-inflammatory, antioxidant, and telomere protection," which can be used to prepare cosmetic skincare products in the anti-aging field. Regarding inflammation regulation, this cyclic heptapeptide can bind to multiple sites on the A-box functional domain of the pro-inflammatory cytokine high-mobility group box 1 (HMGB1), including alanine at position 17, arginine at position 10, and glycine at position 11. This binding induces an allosteric change in HMGB1, inhibiting its release, inhibiting its binding to Toll-like receptor 4 (TLR4), inhibiting its binding to the receptor for advanced glycation end products (RAGE), and blocking the activation of NF-κB in downstream nuclear factor-activated B cells. Regarding antioxidant regulation, this cyclic heptapeptide can competitively bind to Kelch-like ECH-associated protein 1 (Keap1), activating the expression of nuclear factor E2-associated factor 2 (Nrf2), thereby exerting its antioxidant effect. In terms of telomere protection, this cyclic heptapeptide can promote the expression of telomere repeat binding factor 1 (TRF1), thus playing a protective role for telomeres.

[0030] This cyclic heptapeptide can be used to prepare a variety of cosmetic and beauty care products, including essence water, toner, lotion, face cream, beauty balm, foundation, concealer, primer, setting powder, eyeshadow, contouring, blush, lipstick, single-use ampoules, freeze-dried powder, freeze-dried liquid, facial mask liquid, facial mask powder, frozen mask, and functional medical aesthetic products. These products can fully utilize the anti-inflammatory, antioxidant, and telomere-protecting effects of this cyclic heptapeptide to achieve skin care and beauty effects. Example 2

[0031] This embodiment provides a cyclic heptapeptide structure with the amino acid sequence of cyclic Gly-Tyr-Tyr-Glu-Leu-Asn-Asp, an amidation (-NH2) at the C-terminus, and the molecular formula of C39H50N8O14.

[0032] This cyclic heptapeptide was prepared by solid-phase synthesis. The specific preparation method is as follows: First, Rink Amide resin was selected as the solid support, with a resin loading of 0.5-0.8 mmol / g. The resin was immersed in dichloromethane for 30 minutes to allow it to fully swell. Then, the resin was treated twice with a 20% piperidine / N,N-dimethylformamide (DMF) solution for 30 minutes each time to remove the Fmoc protecting groups.

[0033] Subsequently, Fmoc-Asp(OtBu)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Leu-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Tyr(tBu)-OH, and Fmoc-Gly-OH were coupled to the resin sequentially from the C-terminus to the N-terminus. Each coupling reaction used 3 mol of amino acid, 3 mol of HBTU (O-benzotriazole-N,N,N',N'-tetramethylureonium hexafluorophosphate), 3 mol of HOBt (1-hydroxybenzotriazole), and 6 mol of DIEA (N,N-diisopropylethylamine), reacting in DMF solution for 2 hours. After each coupling, the resin was treated twice with 20% piperidine / DMF solution for 20 minutes each time to remove the Fmoc protecting groups.

[0034] After the linear heptapeptide synthesis was completed, the resin was treated with a 1% trifluoroacetic acid / dichloromethane solution to cleave the peptide from the resin while retaining the side chain protecting groups. The obtained linear peptide was dissolved in dichloromethane (DCM), and 1.2 mol of HATU (O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylureon hexafluorophosphate) and 2.4 mol of N,N-diisopropylethylamine (DIEA) were added. The reaction was carried out under dilution conditions (peptide concentration of 1-2 mM) for 24-48 hours to achieve intramolecular cyclization.

[0035] After the cyclization reaction was completed, the mixture was treated with a mixed solution of 95% trifluoroacetic acid, 2.5% triisopropylsilane, and 2.5% water for 4 hours to remove all side-chain protecting groups. The reaction solution was precipitated with cold diethyl ether, the precipitate was collected by centrifugation, and washed three times with diethyl ether.

[0036] The crude product was purified by semi-preparative high-performance liquid chromatography (HPLC) using a C18 reversed-phase column and a water-acetonitrile gradient (containing 0.1% trifluoroacetic acid) as the mobile phase. The purified product was then freeze-dried to obtain the target cyclic heptapeptide as a white powder.

[0037] The molecular weight of this cyclic heptapeptide is 846.87 Da, and its molecular formula was confirmed by high-resolution mass spectrometry (HRMS) analysis to be C64-C ... 39 H 50 N8O 14 Amino acid sequence analysis confirmed it to be a cyclic Gly-Tyr-Tyr-Glu-Leu-Asn-Asp, with an amidation (-NH2) at the C-terminus. (See attached image.) Figure 1 .

[0038] In a preferred embodiment, the purity of the cyclic heptapeptide is greater than 95%, as confirmed by HPLC analysis. The cyclic heptapeptide exhibits good stability under physiological pH conditions (pH 7.4), with a degradation rate of less than 5% after 72 hours of incubation in phosphate buffer at 37°C. Example 3

[0039] This embodiment provides a method for preparing a cyclic heptapeptide. The amino acid sequence of the cyclic heptapeptide is cyclic Gly-Tyr-Tyr-Glu-Leu-Asn-Asp, with an amidation (-NH2) at the C-terminus. The molecular formula is C 39 H 50 N8O 14 This includes the following steps: Step 1: Solid-phase synthesis of chain-like peptide resin 2-Chlorotriphenylmethylchloro (CTC) resin was swollen with dichloromethane (DCM), and amino acids were sequentially coupled starting with Asp to form a chain-like peptide resin. Specifically, Asp was first attached to the resin, followed by deprotection, and then subsequent amino acids were activated with O-benzotriazole-tetramethylurea hexafluorophosphate (HBTU) and coupled sequentially. The deprotection process was carried out using piperidine (PIP) and its derivatives, N,N-dimethylformamide (DMF), and / or trifluoroacetic acid (TFA) solutions.

[0040] Step 2: Cleavage of chain peptide resin The linear peptide crude product is obtained by cleaving the chain peptide resin using 10%–40% hexafluoroisopropanol (HFIP) / dichloromethane (DCM). In a preferred embodiment, cleavage can be performed using 20% ​​hexafluoroisopropanol (HFIP) / DCM for 2 hours.

[0041] Step 3: Purify linear peptides The crude linear peptide was purified by reversed-phase high-performance liquid chromatography (HPLC) to obtain the linear peptide. A C18 column was used during purification, with a water-acetonitrile mobile phase, gradient elution, and a detection wavelength of 220 nm.

[0042] Step 4: Cyclicizing linear peptides The linear peptide was dissolved in the organic solvent N,N-dimethylformamide (DMF), and then 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) and N,N-diisopropylethylamine (DIEA) were added to initiate a cyclization reaction. The reaction process was monitored using liquid chromatography-mass spectrometry (LCMS). The reaction was terminated when the starting peak disappeared and the target product peak appeared and stabilized, yielding the crude cyclized peptide.

[0043] The preferred conditions for the cyclization reaction are: a linear peptide:HATU:DIEA molar ratio of 1:0.3~0.4:0.6~0.8, a reaction temperature of room temperature, and a reaction time of 4-8 hours.

[0044] Step 5: Purification of the finished product The crude cyclic peptide was subjected to cleavage of byproducts, washing, high-performance liquid chromatography (HPLC) purification, and lyophilization to obtain the cyclic heptapeptide product. Specifically, the crude cyclic peptide was first treated with trifluoroacetic acid (TFA) solution to cleave byproducts, then washed with diethyl ether, purified by reversed-phase HPLC, and finally lyophilized to obtain the cyclic heptapeptide product.

[0045] During the pyrolysis of byproducts, a mixed solution of 95% TFA / 2.5% triisopropylsilane / 2.5% water was used for 2 hours. HPLC purification was performed using a C18 column with a water-acetonitrile mobile phase and a detection wavelength of 220 nm. During lyophilization, the sample solution was pre-frozen to -80°C, then primary dried at -50°C, and finally secondary dried at 20°C to obtain a cyclic heptapeptide product with a purity greater than 95%, which was confirmed by LC-MS / MS analysis. Figure 1 and Figure 2 .

[0046] The cyclic heptapeptide prepared by the above method has high purity and yield, and its structure is stable, making it suitable for subsequent drug development and bioactivity research. Example 4

[0047] This embodiment discloses a cyclic heptapeptide with the amino acid sequence cyclic Gly-Tyr-Tyr-Glu-Leu-Asn-Asp, an amidation (-NH2) at the C-terminus, and the molecular formula C 39 H 50 N8O 14 This cyclic heptapeptide possesses inflammatory regulatory, antioxidant regulatory, and telomere protective effects, forming a synergistic anti-aging network of anti-inflammatory, antioxidant, and telomere protective properties, which can be used to prepare cosmetic skincare products in the field of anti-aging.

[0048] This cyclic heptapeptide exerts its inflammatory regulatory effects through multiple mechanisms. Specifically, it binds to multiple sites on the A-box protein functional domain of the pro-inflammatory cytokine high-mobility group box 1 (HMGB1). This A-box functional domain consists of approximately 79 amino acid residues and has a specific three-dimensional spatial structure. Binding of the cyclic heptapeptide to the A-box functional domain induces an allosteric change in HMGB1, thereby inhibiting its release. Furthermore, the cyclic heptapeptide also inhibits the binding of HMGB1 to Toll-like receptor 4 (TLR4) and simultaneously inhibits the binding of HMGB1 to the receptor for advanced glycation end products (RAGE) target, thereby blocking the activation of NF-κB in downstream nuclear factor-activated B cells.

[0049] In a preferred embodiment, the cyclic heptapeptide binds to specific sites on the A-box domain of HMGB1, including alanine at position 17, arginine at position 10, and glycine at position 11. This binding to these sites is the molecular basis for its inflammatory regulatory function.

[0050] This cyclic heptapeptide also exhibits significant antioxidant regulatory effects. By competitively binding to Kelch-like ECH-associated protein 1 (Keap1), the cyclic heptapeptide activates the expression of nuclear factor E2-associated factor 2 (Nrf2), thereby initiating the intracellular antioxidant defense system. Keap1 is a negative regulator of Nrf2; under normal conditions, Keap1 binds to Nrf2, causing it to remain in the cytoplasm and promoting its degradation. After the cyclic heptapeptide binds to Keap1, Nrf2 is released from the Keap1-Nrf2 complex. Subsequently, Nrf2 translocates to the nucleus, activating antioxidant response element (ARE)-mediated expression of antioxidant genes, including antioxidant enzymes such as glutathione peroxidase and superoxide dismutase, thereby enhancing the cell's antioxidant capacity.

[0051] Furthermore, this cyclic heptapeptide also has telomere protective effects. It can promote the expression of telomere repeat-binding factor 1 (TRF1). TRF1 is an important component of the telomere protein complex, specifically binding to telomeric DNA repeat sequences to maintain the integrity of telomere structure. By promoting TRF1 expression, the cyclic heptapeptide can enhance telomere stability, slow down telomere shortening, and thus delay the cellular senescence process.

[0052] The cyclic heptapeptide's synergistic effects of inflammation regulation, antioxidant regulation, and telomere protection form a complete anti-aging network. Inflammation and oxidative stress are crucial factors in cellular senescence, with telomere shortening being a hallmark event of this process. The cyclic heptapeptide alleviates inflammation by inhibiting the HMGB1-mediated inflammatory pathway; reduces oxidative stress levels by activating the Nrf2-mediated antioxidant pathway; and protects telomere integrity by promoting TRF1 expression. These three mechanisms work synergistically to combat cellular senescence, forming a highly effective anti-aging network.

[0053] This synergistic mechanism of action across multiple targets and mechanisms makes the cyclic heptapeptide a promising candidate for application in the field of anti-aging. By simultaneously regulating inflammatory responses, oxidative stress, and telomere protection, the cyclic heptapeptide can comprehensively delay the cellular aging process, providing new ideas and directions for the development of anti-aging drugs and health products. Example 5

[0054] The inflammatory regulatory mechanism of this cyclic heptapeptide involves binding to the A-box domain of the pro-inflammatory cytokine high-mobility group box 1 (HMGB1), thereby modulating the inflammatory response. This cyclic heptapeptide can bind to multiple sites on the A-box domain of HMGB1, including alanine at position 17, arginine at position 10, and glycine at position 11. (See...) Figure 3 .

[0055] The inflammatory regulatory mechanism of this cyclic heptapeptide involves binding to the A-box domain of HMGB1, causing a conformational change in HMGB1. The A-box domain consists of approximately 79 amino acid residues and has a specific three-dimensional spatial structure. Upon binding to multiple key sites on this domain, the cyclic heptapeptide can alter the spatial conformation of HMGB1, thereby affecting its biological function.

[0056] In a preferred embodiment, the cyclic heptapeptide binds to alanine residue at position 17 of the A-box domain of HMGB1. This binding site plays a crucial role in the allosteric changes in HMGB1 induced by the cyclic heptapeptide. Through interaction with this site, the cyclic heptapeptide can be stably anchored to the HMGB1 molecule, thereby triggering a series of conformational changes.

[0057] In another preferred embodiment, the cyclic heptapeptide binds simultaneously to both arginine at position 10 and glycine at position 11 of the A-box domain of HMGB1. This synergistic binding at these two sites further enhances the interaction between the cyclic heptapeptide and HMGB1, enabling the cyclic heptapeptide to more effectively induce allosteric changes in HMGB1.

[0058] This cyclic heptapeptide, by inducing an allosteric change in HMGB1, inhibits its release. In inflammatory states, HMGB1 typically translocates from the nucleus to the cytoplasm and is eventually released into the extracellular space, exerting a pro-inflammatory effect. By binding to HMGB1 and inducing an allosteric change, the cyclic heptapeptide prevents HMGB1 from translocating from the nucleus to the cytoplasm, thereby inhibiting its release into the extracellular space. Figure 4 As shown, the inventor used 9 J / cm 2 A human dermal fibroblast model was established by UVA irradiation. After modeling, the cells were administered drugs and incubated with heptapeptide-6 and cyclic heptapeptide-6 for 24 hours, respectively. The solutions were then discarded, followed by washing, fixation, permeabilization, and antibody incubation. Fluorescence microscopy was used to photograph the cells. The results showed that when the concentration of cyclic heptapeptide was above 10 ppm, it could effectively inhibit the release of HMGB1, which was stronger than heptapeptide-6 in this study.

[0059] Cyclic heptadipeptide also inhibits the binding of HMGB1 to Toll-like receptor 4 (TLR4). Under normal conditions, HMGB1 released extracellularly can bind to TLR4, activating downstream signaling pathways and promoting inflammatory responses. Cyclic heptadipeptide-induced allosteric changes in HMGB1 alter the spatial conformation of the HMGB1-TLR4 binding site, preventing HMGB1 from effectively binding to TLR4 and thus inhibiting HMGB1-mediated TLR4 signaling pathway activation.

[0060] The inventors' research indicates that the competitive binding affinity of this cyclic heptapeptide to TLR4 is approximately -0.8 kcal / mol. Figure 5 As shown, in in vitro experiments, after treating keratinocytes stimulated by recombinant HMGB1 protein with 10 ppm cyclic heptapeptide for 24 hours, the intracellular TLR4 gene expression was reduced by 80.89% compared with the control group, indicating that when the cyclic heptapeptide was 10 ppm, it competitively inhibited the TLR4 gene expression. After treating keratinocytes stimulated by recombinant HMGB1 protein with 50 ppm cyclic heptapeptide for 24 hours, the intracellular TLR4 gene expression was reduced by 85.23% compared with the control group, indicating that when the cyclic heptapeptide was 50 ppm, it more effectively competitively inhibited the TLR4 gene expression.

[0061] Furthermore, this cyclic heptapeptide can also inhibit the binding of HMGB1 to the receptor for advanced glycation end products (RAGE). RAGE is another important receptor for HMGB1, and the binding of HMGB1 to RAGE can also activate pro-inflammatory signaling pathways. By binding to HMGB1 and causing a conformational change, the cyclic heptapeptide alters the spatial conformation of the HMGB1-RAGE binding site, preventing HMGB1 from effectively binding to RAGE and thus inhibiting HMGB1-mediated RAGE signaling pathway activation.

[0062] The inventors' research indicates that the competitive binding affinity of this cyclic heptapeptide to RAGE is approximately -6.6 kcal / mol. Figure 6 As shown, in in vitro experiments, after treating keratinocytes stimulated by recombinant HMGB1 protein with 10 ppm cyclic heptapeptide for 24 hours, the intracellular RAGE gene expression was reduced by 56.0% compared with the control group, indicating that when the cyclic heptapeptide was 10 ppm, it competitively inhibited the RAGE gene expression. After treating keratinocytes stimulated by recombinant HMGB1 protein with 50 ppm cyclic heptapeptide for 24 hours, the intracellular RAGE gene expression was reduced by 65.3% compared with the control group, indicating that when the cyclic heptapeptide was 50 ppm, it more effectively competitively inhibited the RAGE gene expression.

[0063] By inhibiting the binding of HMGB1 to TLR4 and RAGE, cyclic heptapeptide can block the activation of the κ-light chain enhancer (NF-κB) in downstream nuclear factor-activated B cells. NF-κB is an important transcription factor that plays a crucial role in the inflammatory response. Under normal circumstances, the binding of HMGB1 to TLR4 or RAGE activates a series of signaling pathways, ultimately leading to NF-κB activation and promoting the expression of pro-inflammatory factors. Cyclic heptapeptide inhibits the binding of HMGB1 to TLR4 and RAGE, blocking the activation of these signaling pathways, thereby inhibiting NF-κB activation, reducing the expression of pro-inflammatory factors, and ultimately achieving the effect of suppressing the inflammatory response.

[0064] In summary, this cyclic heptapeptide's inflammatory regulatory mechanism involves binding to multiple sites on the A-box domain of HMGB1 (including alanine at position 17, arginine at position 10, and glycine at position 11), causing allosteric changes in HMGB1, inhibiting its release, suppressing its binding to TLR4 and RAGE, and blocking the activation of downstream NF-κB, thereby achieving the effect of inhibiting the inflammatory response. Example 6

[0065] This embodiment relates to a cyclic heptapeptide antioxidant and telomere protection mechanism, which achieves competitive binding to Keap1 and promotes the expression of TRF1 through a specific cyclic heptapeptide sequence, thereby activating Nrf2 expression and protecting telomere structure.

[0066] The antioxidant mechanism of this cyclic heptapeptide partly employs a specially designed cyclic heptapeptide structure, in which seven amino acid residues are linked end-to-end by peptide bonds to form a cyclic structure. The amino acid sequence of the cyclic heptapeptide is cyclic Gly-Tyr-Tyr-Glu-Leu-Asn-Asp, with an amidation (-NH2) at the C-terminus, forming a cyclic structure. This cyclic structure endows the peptide chain with high stability and specific conformation, enabling it to accurately recognize and bind to specific binding sites on the Keap1 protein.

[0067] The competitive binding mechanism of this cyclic heptapeptide to Keap1 is based on molecular simulations and experimental verification. Residues such as glutamate (Glu) in the cyclic heptapeptide can form crucial salt bridges or hydrogen bonds with positively charged arginine residues in the Kelch domain of Keap1. When the cyclic heptapeptide binds to Keap1, it occupies the site originally used for binding Nrf2, thereby blocking the interaction between Keap1 and Nrf2. Under normal physiological conditions, Keap1 promotes the ubiquitination and proteasome degradation of Nrf2 through binding to Nrf2. The competitive binding of the cyclic heptapeptide disrupts this process, preventing Nrf2 from being degraded.

[0068] The mechanism by which this cyclic heptapeptide activates Nrf2 expression is as follows: after the cyclic heptapeptide competitively binds to Keap1, the unbound Nrf2 protein remains stable and accumulates, subsequently being transported to the cell nucleus. In the nucleus, Nrf2 forms a heterodimer with small Maf protein, which binds to antioxidant response elements (AREs), initiating the transcription of downstream antioxidant genes, including glutathione synthase, heme oxygenase-1, and NAD(P)H quinone oxidoreductase 1. The expression of these antioxidant enzymes enhances the cell's defense against oxidative stress.

[0069] The inventors' research indicates that the competitive binding affinity of this cyclic heptapeptide to Nrf2 is approximately -3.4 kcal / mol. Figure 7 As shown, in in vitro experiments, after treating fibroblasts with 10 ppm of cyclic heptapeptide for 24 hours, Nrf2 expression was quantitatively detected by ELISA. Intracellular Nrf2 gene expression increased by +13.42% compared to the control group, indicating that 10 ppm of cyclic heptapeptide promoted Nrf2 gene expression. After treating fibroblasts with 50 ppm of cyclic heptapeptide for 24 hours, Nrf2 gene expression increased by +16.38% compared to the control group, indicating that 50 ppm of cyclic heptapeptide better promoted Nrf2 gene expression. In contrast, the competitive binding ability of heptapeptide-6 to Nrf2 was approximately -5.0 kcal / mol; when heptapeptide-6 was 10 ppm, it promoted Nrf2 gene expression by +12.39%; when heptapeptide-6 was 50 ppm, it promoted Nrf2 gene expression by +15.18%, both of which were inferior to cyclic heptapeptide in this study.

[0070] In the oxidative stress model, pretreatment with cyclic heptapeptide increased cell survival by 65% ​​and reduced intracellular reactive oxygen species (ROS) levels by 58%, indicating that cyclic heptapeptide effectively protects cells from oxidative damage by activating the Nrf2 signaling pathway.

[0071] The telomere protection mechanism of this cyclic heptapeptide is partly achieved by promoting the expression of telomere repeat-binding factor 1 (TRF1). This cyclic heptapeptide can bind to specific intracellular transcription factors, such as SP1 and E2F, enhancing their binding affinity to the TRF1 gene promoter region. The specific mechanism by which the cyclic heptapeptide promotes TRF1 expression involves epigenetic modification regulation. After cyclic heptapeptide treatment, the level of histone acetylation (H3K27ac) in the TRF1 gene promoter region significantly increased, while the level of DNA methylation decreased. These changes facilitate the access of transcription factors to DNA and promote gene transcription. Furthermore, the cyclic heptapeptide can also inhibit the expression of specific miRNAs (such as miR-155), which normally inhibit the translation of TRF1 mRNA; therefore, the effect of the cyclic heptapeptide further enhances TRF1 protein production.

[0072] TRF1 is an important component of the telomere protein complex. Increased expression of TRF1 enables it to bind more effectively to telomeric DNA repeat sequences (TTAGGG)n, protecting telomeres from nuclease degradation. TRF1 can also recruit other telomere protective proteins such as TRF2, POT1, TIN2, TPP1, and Rap1 to form the telomere protection complex (shelterin), maintaining the integrity of telomere structure. In addition, TRF1 is involved in regulating telomere length, preventing excessive elongation or shortening of telomeres, thereby maintaining chromosome stability.

[0073] The inventors' research indicates that the competitive binding affinity of this cyclic heptapeptide to TRF1 is approximately -3.3 kcal / mol. Figure 8 As shown, in senescent cell models, such as the zebrafish model of telomere damage induced by hydrogen peroxide, treatment with 10 ppm cyclic heptapeptide at 28°C in the dark for 120 h increased the TRF1 gene expression level by 103% compared to the control group, indicating that 10 ppm cyclic heptapeptide promoted TRF1 gene expression. After treatment with 50 ppm cyclic heptapeptide for 120 h, the TRF1 gene expression level increased by 262% compared to the control group, indicating that 50 ppm cyclic heptapeptide greatly promoted TRF1 gene expression. This suggests that cyclic heptapeptide effectively delayed the cellular senescence process by promoting TRF1 expression.

[0074] The antioxidant and telomere protection mechanisms of this cyclic heptapeptide exhibit a synergistic effect. The antioxidant effect generated by Nrf2 activation reduces oxidative damage to telomere DNA, while increased TRF1 expression directly enhances the physical protection of telomeres. This dual protective mechanism gives the cyclic heptapeptide a unique advantage in anti-aging and prevention of oxidative stress-related diseases.

[0075] In a preferred embodiment, the cyclic heptacapeptide can be prepared using solid-phase peptide synthesis technology, achieving a purity of over 98%. The cyclic heptacapeptide can be stored as a lyophilized powder, dissolved in PBS or DMSO for use, and exhibits good stability under physiological conditions at 37°C, with a half-life exceeding 24 hours. The cyclic heptacapeptide maintains its activity within a pH range of 5.5-8.5, with an optimal pH of 7.2. The cyclic heptacapeptide can be administered via various routes of administration, including intravenous injection, subcutaneous injection, or oral formulations. Example 7

[0076] As can be seen from the application of the cyclic heptapeptide provided by the present invention in cosmetics and beauty care products, the cyclic heptapeptide can be used to prepare a variety of cosmetics and / or beauty and care products.

[0077] In one embodiment of this invention, the cyclic heptapeptide can be used to prepare essence water or toner products. In this embodiment, the concentration of the cyclic heptapeptide is 0.01%~5%, preferably 0.05%~2%, and more preferably 0.1%~1%. In a specific embodiment, the cyclic heptapeptide is added at a concentration of 0.5% to a matrix containing purified water, glycerin, propylene glycol, sodium hyaluronate, and preservatives, resulting in an essence water with good moisturizing and anti-aging effects.

[0078] The cyclic heptapeptide provided by this invention is used to prepare various cosmetic and / or beauty and skincare products. There are also various implementation methods, such as: This cyclic heptapeptide can be used to prepare emulsion products. In this embodiment, the concentration of the cyclic heptapeptide is 0.01% to 3%, preferably 0.05% to 1.5%, and more preferably 0.1% to 0.8%. In a preferred embodiment, the cyclic heptapeptide is added at a concentration of 0.3% to an emulsion system containing an aqueous phase and an oil phase, wherein the aqueous phase includes purified water, glycerin, propylene glycol, and sodium hyaluronate, and the oil phase includes vegetable oil, silicone oil, and an emulsifier. The resulting emulsion has good moisturizing effects and skin-firming functions.

[0079] This cyclic heptapeptide can be used to prepare face cream products. In this embodiment, the concentration of the cyclic heptapeptide is 0.01% to 4%, preferably 0.1% to 2%, and more preferably 0.2% to 1%. In one specific embodiment, the cyclic heptapeptide is added at a concentration of 0.8% to an emulsion system containing an aqueous phase and an oil phase, wherein the oil phase content is higher than that of the emulsion, resulting in a face cream with stronger moisturizing and repairing effects, suitable for nighttime use.

[0080] This cyclic heptapeptide can be used to prepare beauty cream products. In this embodiment, the concentration of the cyclic heptapeptide is 0.05% to 5%, preferably 0.1% to 3%, and more preferably 0.5% to 2%. In a preferred embodiment, the cyclic heptapeptide is added at a concentration of 1.5% to a matrix containing petrolatum, beeswax, glycerin, and vegetable oil, and the resulting beauty cream has good repairing and moisturizing effects.

[0081] This cyclic heptapeptide can be used to prepare base makeup products. In this embodiment, the concentration of the cyclic heptapeptide is 0.01% to 2%, preferably 0.05% to 1%, and more preferably 0.1% to 0.5%. In one specific embodiment, the cyclic heptapeptide is added at a concentration of 0.2% to a matrix containing water, silicone oil, pigment, filler, and emulsifier. The resulting base makeup product not only has good concealing effect but also provides skin care benefits during use.

[0082] This cyclic heptapeptide can be used to prepare concealer products. In this embodiment, the concentration of the cyclic heptapeptide is 0.01% to 3%, preferably 0.05% to 1.5%, and more preferably 0.1% to 0.8%. In a preferred embodiment, the cyclic heptapeptide is added at a concentration of 0.4% to a matrix containing a high concentration of pigment, silicone oil, and binder, resulting in a concealer product with good coverage and skin care effects.

[0083] This cyclic heptapeptide can be used to prepare isolation products. In this embodiment, the concentration of the cyclic heptapeptide is 0.01% to 2.5%, preferably 0.05% to 1.2%, and more preferably 0.1% to 0.6%. In one specific embodiment, the cyclic heptapeptide is added at a concentration of 0.3% to a matrix containing titanium dioxide, zinc oxide, silicone oil, and an emulsifier, and the resulting isolation product has good protective and repairing effects.

[0084] This cyclic heptapeptide can be used to prepare setting products. In this embodiment, the concentration of the cyclic heptapeptide is 0.01% to 2%, preferably 0.05% to 1%, and more preferably 0.1% to 0.5%. In a preferred embodiment, the cyclic heptapeptide is added at a concentration of 0.2% to a matrix containing talc, silica, and trace amounts of oil, resulting in a setting product with good oil control and skin care functions.

[0085] This cyclic heptapeptide can also be used to prepare eyeshadow products. In this embodiment, the concentration of the cyclic heptapeptide is 0.01% to 3%, preferably 0.05% to 1.5%, and more preferably 0.1% to 0.8%. In one specific embodiment, the cyclic heptapeptide is added at a concentration of 0.3% to a matrix containing pigments, mica powder, silicone oil, and binders, resulting in an eyeshadow product with good coloring effect and eye care function.

[0086] This cyclic heptapeptide can also be used to prepare contouring products. In this embodiment, the concentration of the cyclic heptapeptide is 0.01% to 2.5%, preferably 0.05% to 1.2%, and more preferably 0.1% to 0.6%. In a preferred embodiment, the cyclic heptapeptide is added at a concentration of 0.4% to a matrix containing pigments, fillers, and binders, resulting in a contouring product with good three-dimensional shaping effect and skin care function.

[0087] This cyclic heptapeptide can also be used to prepare blush products. In this embodiment, the concentration of the cyclic heptapeptide is 0.01% to 2%, preferably 0.05% to 1%, and more preferably 0.1% to 0.5%. In one specific embodiment, the cyclic heptapeptide is added at a concentration of 0.3% to a matrix containing organic pigments, fillers, and binders, resulting in a blush product with a natural coloring effect and skin-repairing function.

[0088] This cyclic heptapeptide can also be used to prepare lipstick products. In this embodiment, the concentration of the cyclic heptapeptide is 0.01% to 3%, preferably 0.05% to 1.5%, and more preferably 0.1% to 0.8%. In a preferred embodiment, the cyclic heptapeptide is added at a concentration of 0.5% to a matrix containing beeswax, vegetable oil, pigment, and silicone oil, resulting in a lipstick product with good moisturizing and lip-repairing effects.

[0089] This cyclic heptapeptide can also be used to prepare single-use ampoule products. In this embodiment, the concentration of the cyclic heptapeptide is 0.1% to 10%, preferably 0.5% to 5%, and more preferably 1% to 3%. In one specific embodiment, the cyclic heptapeptide is added at a concentration of 2% to a matrix containing purified water, propylene glycol, sodium hyaluronate, and preservatives, resulting in a single-use ampoule with highly effective skin repair and anti-aging effects.

[0090] This cyclic heptapeptide can also be used to prepare lyophilized powder or lyophilized liquid products. In this embodiment, the concentration of the cyclic heptapeptide is 0.5% to 15%, preferably 1% to 10%, and more preferably 2% to 7%. In a preferred embodiment, the cyclic heptapeptide is mixed at a concentration of 5% with sodium hyaluronate, collagen, and preservatives, and then lyophilized to obtain a lyophilized powder with excellent stability and highly effective skin repair function. It can be mixed with a special solvent and applied directly to the skin before use.

[0091] This cyclic heptapeptide can also be used to prepare facial mask liquids, facial mask powders, or gel masks. In this embodiment, the concentration of the cyclic heptapeptide is 0.05% to 8%, preferably 0.1% to 4%, and more preferably 0.5% to 2%. In one specific embodiment, the cyclic heptapeptide is added at a concentration of 1% to a matrix containing purified water, glycerin, sodium hyaluronate, collagen, and preservatives, resulting in a facial mask liquid with good moisturizing and firming effects.

[0092] This cyclic heptapeptide can also be used to prepare functional medical aesthetic products. In this embodiment, the concentration of the cyclic heptapeptide is 0.1% to 20%, preferably 0.5% to 10%, and more preferably 1% to 5%. In a preferred embodiment, the cyclic heptapeptide is added at a concentration of 3% to a matrix containing sodium hyaluronate, glycerin, vitamin C, and vitamin E, resulting in a functional medical aesthetic product with significant anti-aging, skin-firming, and damaged skin-repairing effects. This product is suitable for use in medical aesthetic institutions for post-treatment repair.

[0093] Through the application of various cosmetic and beauty care products, this cyclic heptapeptide exhibits excellent skin repair, anti-aging, firming, and moisturizing effects, meeting the diverse needs of different consumers.

[0094] The above are merely specific embodiments of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A cyclic heptapeptide, characterized in that, The amino acid sequence of the cyclic heptapeptide is cyclic Gly-Tyr-Tyr-Glu-Leu-Asn-Asp, with an amidation (-NH2) at the C-terminus, and the molecular formula is C 39 H 50 N8O 14 .

2. The cyclic heptapeptide as described in claim 1, characterized in that, The cyclic heptapeptide is prepared by solid-phase synthesis of peptide chains to obtain chain peptide resin, cleavage to obtain crude linear chain peptide, purification to obtain linear chain peptide, cyclization to obtain crude cyclic peptide, and / or purification of the finished product.

3. The cyclic heptapeptide as described in claim 2, characterized in that, The solid-phase synthesis involves swelling 2-chlorotriphenylmethyl chloride (CTC) resin with dichloromethane (DCM) and sequentially coupling amino acids starting with Asp to form a chain-like peptide resin. The cleavage includes cleaving the chain peptide resin with 10%~40% hexafluoroisopropanol (HFIP) / dichloromethane (DCM) to obtain a crude linear chain peptide product; The purification process includes purifying the crude product by reversed-phase high-performance liquid chromatography (HPLC) to obtain a linear peptide. The cyclization process involves dissolving the linear peptide in an organic solvent, including N,N-dimethylformamide (DMF) and dichloromethane (DCM), adding 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) and / or N,N-diisopropylethylamine (DIEA) for cyclization, monitoring the reaction, and obtaining the crude cyclized peptide. The purification of the finished product includes cleaving the crude cyclic peptide into pyrolysis byproducts, washing, high-performance liquid chromatography purification and / or freeze drying to obtain the cyclic heptapeptide.

4. The cyclic heptapeptide as described in claim 3, characterized in that, The coupled amino acid process includes linking Asp to the resin, followed by deprotection and activation with O-benzotriazole-tetramethylurea hexafluorophosphate (HBTU); the deprotection uses a solution of piperidine (PIP) and its derivatives, N,N-dimethylformamide (DMF), and / or trifluoroacetic acid (TFA) as the deprotecting agent; the cyclization process is monitored using liquid chromatography-mass spectrometry (LCMS); the byproducts in the purification of the final product are cleaved using a solution of trifluoroacetic acid (TFA) as the cleavage agent.

5. The cyclic heptapeptide as described in claim 1, characterized in that, The cyclic heptapeptide has inflammatory regulation, antioxidant regulation, and / or telomere protection effects, forming a synergistic anti-aging network of "anti-inflammatory-antioxidant-telomere protection", which can be used to prepare beauty and skin care products in the field of anti-aging.

6. The cyclic heptapeptide as described in claim 5, characterized in that, The inflammatory regulatory effects include binding to multiple sites of the pro-inflammatory cytokine high mobility group box 1 (HMGB1), said sites being located on a protein functional domain (A-box) with a specific three-dimensional spatial structure and consisting of approximately 79 amino acid residues; causing allosteric changes in HMGB1; inhibiting the release of HMGB1; and / or inhibiting the binding of HMGB1 to Toll-like receptor 4 (TLR4); and / or inhibiting the binding of HMGB1 to the receptor for advanced glycation end products (RAGE) target; and / or blocking the activation of κ-light chain enhancement (NF-κB) in downstream nuclear factor-activated B cells.

7. The cyclic heptapeptide as described in claim 6, characterized in that, The substance binds to multiple sites on the high-mobility group box 1 (HMGB1), including alanine at position 17, arginine at position 10, and / or glycine at position 11.

8. The cyclic heptapeptide as described in claim 5, characterized in that, The antioxidant regulation includes competitive binding to Kelch-like ECH-associated protein 1 (Keap1) and / or activation of nuclear factor E2-associated factor 2 (Nrf2) expression.

9. The cyclic heptapeptide as described in claim 5, characterized in that, The telomere protection effect includes promoting the expression of telomere repeat binding factor 1 (TRF1).

10. The cyclic heptapeptide as described in claim 1, characterized in that, The cyclic heptapeptide can be used to prepare cosmetic and / or beauty and care products, including essence water, toner, lotion, face cream, beauty cream, base makeup, concealer, primer, setting makeup, eyeshadow, contouring, blush, lipstick, single-use ampoule, freeze-dried powder, freeze-dried liquid, facial mask liquid, facial mask powder, frozen mask, and / or functional medical beauty products.

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