A recombinant collagen skin barrier repair composition potentiated with an ectoine

Abstract

The present application relates to the technical field of biological medicine material and tissue damage repair preparation, and discloses a recombinant collagen skin barrier repair composition with the effect of ectoine, which comprises recombinant type III collagen, ectoine and deionized water as the balance in percentage by mass; the recombinant type III collagen is composed of components with specific proportions of long and short chains; the hydrated matrix is constructed by stirring the ectoine aqueous solution, and the collagen is added to anchor the side chain of the protein peptide chain in the matrix to form a linear extension conformation; the present application utilizes the physical steric effect of the ordered hydration structure induced by ectoine to block the environmental ion from stripping the surface-bound water of the protein, maintains the conformational stability of the collagen under the high ion strength of the inflammatory exudate, forms a dense net structure at the skin interface, and further reduces the transdermal water loss rate and enhances the residence of the active components at the damaged interface.

Description

Technical Field

[0001] This invention relates to a recombinant collagen skin barrier repair composition enhanced with ectoine, belonging to the technical field of biomedical materials and tissue damage repair agents. Background Technology

[0002] Currently, recombinant collagen utilizes molecular hydration and a physical membrane layer formed on the surface of the stratum corneum to repair the skin barrier. It also acts as a bioactive scaffold to induce endogenous cell adhesion and migration, playing a key role in accelerating the healing of pathological wounds and the reconstruction of the extracellular matrix, and constructing a physical shielding network at the damaged interface. After laser treatment or in cases of acute injury, inflammatory exudates exist at the skin interface, leading to an increase in the local environmental ionic strength. The peptide chains of recombinant collagen molecules are sensitive to environmental charges. Sodium and chloride ions erode the hydration layer on the protein surface, inducing spontaneous aggregation and collapse of the protein peptide chains. They cannot be linearly laid out at the damaged interface. Increasing the protein concentration will exacerbate molecular aggregation due to the high ionic strength environment, generating a discontinuous membrane layer with microporous structures, weakening the repair barrier and the anchoring force of the stratum corneum.

[0003] While existing technologies include formulations combining ectoine and recombinant collagen, such as the Chinese invention patent with authorization announcement number CN117598927B which discloses an anti-wrinkle composition, anti-wrinkle serum, and its preparation method, this approach focuses on the multi-component pharmacological superposition in conventional anti-wrinkle scenarios. However, it does not address the issue of protein conformational stability in high-ionic-strength inflammatory environments. The components lack specific mass ratio constraints and hydration matrix construction processes, failing to form a hydration shell that resists ionic ablation and physical steric hindrance. Furthermore, it does not design a long-short chain gradation for the molecular weight distribution of collagen. When the composition comes into contact with electrolyte-rich inflammatory exudates from the skin, the active proteins are prone to charge neutralization and aggregation, making it difficult to construct a stable, dense, and continuous biological repair network at the damaged interface.

[0004] Therefore, the technical problem to be solved by this invention is how to ensure the linear topological arrangement of active macromolecules in a complex ionic environment by constructing an interface hydration lock-in mechanism, thereby ensuring the sustained release and signal transduction of recombinant protein biological activity and improving the wound-inducing repair and drug delivery capabilities of biopharmaceutical compositions in extreme inflammatory scenarios. Summary of the Invention

[0005] To address the problems mentioned in the background art, the technical solution of the present invention is as follows: an ectoine-enhanced recombinant collagen skin barrier repair composition, comprising the following components by weight percentage:

[0006] Reorganization Type II collagen to ;

[0007] Ikdoin to ;

[0008] Deionized water is used as a reserve to make up the difference. ;

[0009] Among them, restructuring The mass ratio of type 3 collagen to ectoine is: to ;

[0010] Reorganization Type II collagen consists of collagen with a weight-average molecular weight of to The long-chain components and weight-average molecular weight are to It consists of short-chain components, and the mass ratio of long-chain components to short-chain components is [missing information]. to ;

[0011] The composition is prepared by the following steps:

[0012] step Ikdoin in ℃ to Dissolves in deionized water at ℃ and is used as to Stirring speed to A hydrated matrix is ​​obtained;

[0013] step ,exist to Under shear conditions, recombinant materials are added to the hydrated matrix. Type II collagen, using fluid shear force to reorganize The peptide side chains of type II collagen are anchored in the hydrated matrix to form a linear extended conformation within the hydrated matrix;

[0014] The composition at a mass fraction of In a sodium chloride solution environment, recombinant The hydration kinetic radius retention rate of type II collagen is not less than ,and Potential fluctuations are less than .

[0015] Preferably, the long-chain components construct a bio-repairing network on the surface of the stratum corneum, and the short-chain components are distributed within the gaps of the bio-repairing network; recombinant Type II collagen forms a hydrated shell in the hydrated matrix with a physical steric hindrance effect. This hydrated shell repels sodium and chloride ions in the sodium chloride solution environment, thus promoting recombination. Type II collagen at an ionic strength of It maintains its active conformation under certain conditions.

[0016] Preferably, it further includes trehalose and sodium hyaluronate, wherein trehalose, sodium hyaluronate, and ectoine constitute a hydration synergistic system; the osmotic pressure of the composition is [value missing]. to Its hydration stability index Satisfy the following formula: ,in, The hydration stability index of the composition; The percentage content of ectoine by mass; For reorganization Percentage of type II collagen by mass; This refers to the percentage content of trehalose by mass. Values to , Values to .

[0017] Preferred, recombinant Type II collagen contains a repeating amino acid sequence in which hydroxyproline sites bind to the hydration matrix via a hydrogen bond network, enabling the composition to form a cross-linked hydration repair film at the skin interface.

[0018] Preferably, the purity of ectoine is not less than Its mass fraction is aqueous solution in The conductivity at ℃ is lower than Reorganization The isoelectric point of type II collagen is to .

[0019] Preferred steps China joins the restructuring The rate of collagen production is to By coordinating fluid shear force with the dipole moment of ectoin, recombination is guided. The peptide chains of type II collagen complete a linear topological arrangement.

[0020] Preferably, the composition has a mass fraction of Under stress from sodium chloride solution, recombination The hydration kinetic radius of type II collagen remains at to Within the range.

[0021] Preferably, the composition does not contain nonionic surfactants, and the sum of the mass percentages of all components in the composition is [missing information]. .

[0022] Preferred, recombinant The surface of the peptide chain of type II collagen is bound to three continuous water molecule hydration membranes at polar sites. The interlayer hydrogen bond energy of the water molecule hydration membranes is higher than that of the bulk water, which is used to block the erosion of the bound water on the peptide chain surface by environmental ions.

[0023] Preferably, after the composition is applied to the surface of the damaged skin, it forms a thickness of [thickness value missing] from the long-chain component and the short-chain component. to A monolayer membrane.

[0024] Compared with the prior art, the beneficial effects of the present invention are:

[0025] 1. In the repair of the skin barrier with biopharmaceutical-grade recombinant collagen, a specific ratio of ectoine is used to generate an ordered water cluster shielding layer with quasi-lattice characteristics at the polar amino acid residue sites of recombinant type III collagen. This shielding layer can significantly improve the biochemical stability of recombinant protein. The shielding layer repels sodium ions and chloride ions in the inflammatory environment through physical steric hindrance and electrostatic shielding effect, maintains the linear extension conformation of the protein peptide chain at the skin interface, and avoids spontaneous aggregation or collapse inactivation of recombinant type III collagen due to high ionic strength erosion of the hydration layer, thus ensuring its bioavailability as a tissue regeneration template.

[0026] 2. The specific molecular weight distribution of long and short chain groups is repeated, so that the long chain components can build a large-area repair network on the surface of the stratum corneum under the protection of the ordered water cluster shielding layer, while the short chain components fill the micro gaps in the stratum corneum. With the help of specific shear force to induce linear topological arrangement, a high-density biohydration barrier is formed at the damaged skin interface, which enhances the barrier repair ability.

[0027] 3. The pre-hydration process constructs an ordered water cluster matrix, providing a physical template for the directional arrangement of recombinant type III collagen. By utilizing the fluid shear field in synergistic ectoin strong dipole moment induction, the protein peptide chain is transformed from a random coiled state to a stable hydration-locked state, thereby improving the retention rate of the hydration kinetic radius of the composition under simulated inflammatory exudation conditions.

[0028] 4. This invention utilizes an ordered water cluster matrix induced by ectoin to exhibit a universal synergistic mechanism across components. Through a preferential repulsion effect driven by a strong dipole moment, it can physically repel environmental electrolytes outside the solvation layer of bioactive macromolecules such as sodium hyaluronate, solving the problem of conformational curling and hydration volume collapse of sodium hyaluronate in a high-salt inflammatory exudative environment. This composite hydration mechanism realizes the nonlinear superposition of the structural repair function of recombinant collagen and the biofilm-forming function of sodium hyaluronate, forming an adaptive repair template with high stress resistance, improving the interfacial retention and barrier remodeling accuracy of the composition in complex pathological scenarios. Attached Figure Description

[0029] Figure 1 This is a flowchart illustrating the component structure and stepwise induced preparation process of the composition of the present invention;

[0030] Figure 2 This is a characteristic curve showing the effect of feeding rate on viscosity under a fluid shear field according to the present invention.

[0031] Figure 3 This is a schematic diagram of the protein conformation lock-in and barrier repair mechanism in the inflammatory scenario of this invention. Detailed Implementation

[0032] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0033] This invention provides an ectoin-enhanced recombinant collagen skin barrier repair composition. Through specific component physicochemical interactions and preparation processes, a multiphase hydration system capable of maintaining protein secondary structure stability under high ionic strength environments is constructed. This composition can serve as a core component of medical wound dressings or bioactive scaffolds. Its core architecture utilizes ectoin as a regulator of water molecule structure, inducing the formation of long-range ordered tetrahedral water molecule clusters in a solvent environment. These ordered water clusters are then used as physical templates to spatially confine and protect the peptide chain conformation of recombinant type III collagen. The technical solution of this invention addresses the problem of traditional recombinant collagen failing to respond to inflammatory exudates (which typically contain...) When sodium chloride and various electrolytes are present at certain concentrations, the surface bound water layer is eroded by ions, leading to the failure of charge shielding. This results in the spontaneous curling, aggregation, and precipitation of protein molecules. This can be addressed by controlling the mass ratio of recombinant type III collagen to ectoine within a specific range of 1:4 to 1:10. to Within the isotonic or near-isotonic osmotic pressure window, an environment capable of physically repelling environmental sodium ions is established. ) and chloride ions ( The supramolecular hydration shell ensures that recombinant type III collagen maintains a linearly extended topological conformation at the damaged skin interface, thereby effectively filling and repairing the microscopic gaps in the stratum corneum.

[0034] To address the issues of barrier function loss and loose stratum corneum structure in damaged skin interfaces, single-molecule collagen often struggles to simultaneously achieve both surface film continuity and deep interstitial filling. Therefore, this invention employs a recombinant type III collagen system with a bimodal distribution, consisting of collagen with a weight-average molecular weight of... to The long-chain components and weight-average molecular weight are to It consists of short-chain components, and the mass ratio of long-chain components to short-chain components is limited to a certain value. to Between the components, the long-chain components, utilizing their longer peptide chain spans, form a continuous bio-repairing network along the skin's texture on the surface of the stratum corneum, serving as the main framework of the macroscopic barrier. The short-chain components, with their higher diffusion coefficients and smaller hydrodynamic volumes, penetrate and fill the pores of the bio-repairing network and the microscopic lipid matrix gaps between keratinocytes. This gradation design of long-chain bridging and short-chain filling allows the thickness of the repair layer formed after drying to be controlled within a specific range. to At the monolayer level, it reduces transdermal water loss while ensuring breathability.

[0035] In the specific formulation system, ectoine exists not only as a moisturizer, but also as a physical anchoring agent for protein conformation. On the surface of damaged skin, the high ionic strength of inflammatory exudate can disrupt the hydration layer of proteins, leading to the exposure of hydrophobic groups and causing protein aggregation. To address this challenge, this invention limits the mass percentage of ectoine to [specific percentage missing]. to Furthermore, the mass ratio of recombinant type III collagen to ectoine is [missing information]. to Within this concentration range and ratio, the zwitterionic groups (carboxyl and amino groups on the tetrahydropyrimidine ring) in the ectoin molecule can polarize the surrounding water molecules, causing them to oriented and arrange into a continuous hydrated film of three or more layers. The interlayer hydrogen bond energy is higher than that of bulk water, forming a hydrated matrix with quasi-lattice characteristics. When recombinant type III collagen is dispersed in this matrix, the hydroxyproline and other polar amino acid residues on its peptide chains are anchored to the ordered water clusters in the matrix through a hydrogen bond network. This anchoring produces a microscopic steric hindrance effect, physically blocking free ions in the environment from directly contacting the protein backbone, thereby preventing double-layer compression and protein salting out caused by ionic strength. In this system, even at a mass fraction of [missing information], [missing information]. In a sodium chloride solution environment, the hydration kinetic radius retention rate of recombinant type III collagen is still not lower than And its The amplitude of the potential fluctuation is less than .

[0036] To ensure the formation of the aforementioned microstructure, this invention does not employ a conventional direct mixing process, but rather a stepwise induced preparation procedure. This procedure performs an ordered water cluster pre-construction step (step S101), in which ectoine is... to Dissolved in deionized water under constant temperature conditions, and as to Stirring at a specific speed to This step, through specific thermodynamic conditions and hydrodynamic disturbances, fully solvates ectoin and induces the water molecules in the solvent to rearrange from a disordered state to an ordered cluster structure, forming a hydrated matrix as a carrier, and performing a low-shear segment stretching step (step S102), that is, in to Under low shear conditions, with to Recombinant type III collagen is slowly added to the hydrated matrix at a low rate. During this stage, the low shear rate avoids the destruction of the nascent water bridge connections by strong turbulence. At the same time, the fluid shear force, in conjunction with the dipole moment of ectoin, guides the peptide chains of recombinant type III collagen to unfold along the arrangement direction of the water clusters, and finally forms and locks into a linear extended conformation in the hydrated matrix.

[0037] For implementation schemes requiring further enhancement of environmental adaptability, trehalose and sodium hyaluronate can also be introduced into the composition. Together with ectoine, they form a synergistic hydration system. In this multi-component system, the amount of each component added must meet a specific hydration stability index. The constraint relationship, i.e. ,in, , , These represent the mass percentage content of ectoine, recombinant type III collagen, and trehalose, respectively. As the principal stability coefficient, it takes the value of to This reflects the dominant contribution of ectoin to the protein hydration shell; As an auxiliary coordination coefficient, it takes the value of to This reflects the auxiliary stabilizing effect of trehalose on the glass transition temperature. The value is controlled within a specific range and the overall osmotic pressure of the composition is maintained at [value missing]. to In this synergistic system, the ionic disorder at the damaged interface can be transformed into a trigger signal. Specifically, the higher the ionic strength, the denser the ectoin-induced water cluster structure, thereby enhancing the composition's adaptive protective capability in extreme inflammatory scenarios. In the system of this invention, the ordered water cluster matrix constructed with ectoin induces a universal synergistic mechanism across components. Through its strong dipole moment-driven preferential repulsion effect, it not only plays a crucial anchoring and locking role against the peptide chain conformation of recombinant type III collagen, but also provides environmental stability protection for biomacromolecules such as sodium hyaluronate introduced into the system. Specifically, when this composition is applied to an acutely damaged skin interface with high-salt inflammatory exudation, free sodium and chloride ions in the environment tend to erode the bound water layer on the surface of active macromolecules, leading to a decrease in the density of the hyaluronic acid. Long-chain polysaccharides such as sodium hyaluronate undergo conformational coiling and hydration volume collapse, thereby weakening their original barrier sealing and wetting depth. At this time, the quasi-lattice hydrated matrix generated by ectoin polarization acts as a molecular-level physical shielding layer, excluding environmental electrolytes from the solvation layer of sodium hyaluronate, allowing it to maintain a linearly extended topological state even under high ionic strength. This composite hydration mechanism based on ectoin regulation enables product systems with ectoin combined with collagen, ectoin combined with sodium hyaluronate, or ectoin combined with both to overcome the pharmacological limitations of single components, achieving nonlinear superposition of structural repair function and biological film-forming function, forming an adaptive repair template with high stress resistance, ensuring the interfacial retention and repair accuracy of multiple active components in complex pathological scenarios.

[0038] To ensure that the raw materials meet the requirements for constructing the aforementioned microstructure, this invention strictly limits the physicochemical properties of key components, and the purity of the selected ectoine is not lower than [specific value missing]. And its mass fraction is aqueous solution in The conductivity below needs to be lower than To eliminate interference from impurity ions in the construction of the hydrated matrix, the selected recombinant type III collagen has an isoelectric point (...). ) Controlled within to The composition contains a specific repeating amino acid sequence, in which the density of hydroxyproline sites is sufficient to support the formation of a continuous hydrogen bond network with the hydrated matrix. The composition as a whole does not contain nonionic surfactants to avoid surfactant molecules from intercalating into the protein-water interface and damaging the key hydrated shell structure. Through the systematic integration of the above components, proportions and processes, the present invention finally obtains a skin barrier repair composition that appears as a homogeneous and transparent liquid on a macroscopic scale and has anti-ionic abrasion capabilities on a microscopic scale. This composition has high biocompatibility and can be widely used in the preparation of pharmaceutical preparations or medical devices that promote the repair of skin and soft tissue.

[0039] Example 1: In a clinical nursing scenario of acute repair after phototherapy, the skin barrier is damaged and accompanied by inflammatory exudate with high ionic strength, including sodium ions. Concentration maintained at to And chloride ions Concentration maintained at to This causes local osmotic pressure fluctuations to The above causes conventional collagen preparations to suffer from double-layer compression and molecular aggregation and precipitation due to the erosion of the surface hydration layer, preventing the formation of an effective coating. When the recombinant collagen skin barrier repair composition of this invention, enhanced with ectoine, comes into contact with this damaged interface, the composition contains [amount missing] by mass percentage. to The ectoin molecule, using its dipole moment, polarizes and locks free water molecules in the exudate into ordered water clusters with a tetrahedral structure, thereby constructing a layer with a thickness greater than [missing information] around the recombinant type III collagen molecules. It also possesses a high-barrier hydration shell with a dense hydrogen bond network, which physically repels high concentrations of sodium ions from the environment. With chloride ions It penetrates and blocks the replacement of bound water on the protein surface.

[0040] Protected by this stable hydration shell, the recombinant type III collagen complex maintains a linear extended conformation in an inflammatory environment, with a weight-average molecular weight of [missing information]. to The long-chain components self-assemble along the skin grain direction to form a continuous macroscopic repair network to resist exudate erosion, while the weight-average molecular weight is to The short-chain components, utilizing their low hydrodynamic volume, pass through the pores of the network and fill the lipid matrix gaps between keratinocytes. Together, they form a layer of thickness [missing information] on the wound surface after drying. to A dense monolayer repair film; Continuous monitoring data from a skin moisture meter indicates that the skin area treated with this composition... Inner transdermal water loss rate The reduction exceeded Furthermore, the decrease in the skin surface erythema index confirms that the ectoin-induced hydration shell effectively maintains the micro-conformation stability of recombinant type III collagen and achieves macro-barrier remodeling under extreme physiological conditions.

[0041] Example 2: To verify the technical efficacy of the ectoin-enhanced recombinant collagen skin barrier repair composition of the present invention in real-world application scenarios and to determine the optimal ratio range of the core components, this example designed a gradient control experiment covering multi-dimensional parameter changes. The experiment simulated high ionic strength and inflammatory exudation environments to evaluate the conformational stability, film integrity, and effect on transdermal water loss rate of the composition under extreme conditions. The improvement effect; the experimental platform uses simulated inflammatory exudate as the test medium, prepared from deionized water, which contains a concentration of Sodium chloride Concentration is calcium chloride and concentration of Lactic acid, and through sodium hydroxide The solution will Value adjusted to The overall osmotic pressure is set to To simulate the biochemical environment of damaged skin surface.

[0042] To eliminate random factors and verify the robustness of the technical solution, the experimental design included multiple groups of samples, including the sample group of this invention, a partially missing control group, and an out-of-range control group. The specific composition and test results of each sample are detailed in Table 1. The sample group of this invention was prepared according to the preferred component ratio; control group 1 contained only recombinant type III collagen without ectoine, used to verify the synergistic effect; control group 2, although containing ectoine, had a recombinant type III collagen to ectoine mass ratio set at [value missing]. The ratio is below the lower limit of the range defined in this invention, used to verify the critical significance of the ratio range; the mass ratio of recombinant type III collagen to ectoine in control group 3 is set at [value missing]. The range is higher than the upper limit defined by this invention, and is used to investigate the possible negative effects of excessive ectoine.

[0043] Table 1: Formulation Composition and Key Performance Test Results of Each Sample Group

[0044]

[0045] During the experiment, a dynamic light scattering instrument was used. Monitoring the incubation of each sample group in simulated inflammatory exudate The mean hydration kinetic radius of the protein was determined, and data showed that the retention rates of both sample group A and sample group B exceeded [a certain value]. This indicates that ectoin induces the formation of a stable hydrated shell, effectively resisting ion erosion. In contrast, the retention rate of control group 1 was only [missing information]. This indicates that in the absence of ectoine protection, recombinant collagen aggregates and undergoes conformational collapse. Although the retention rate in control group 2 was improved, it was still lower than that in the sample group of this invention, confirming that... When the mass ratio is below the lower limit, ectoine is insufficient to form a complete hydration network covering the protein; each sample group was uniformly coated onto a simulated skin substrate, and... Films were formed under constant temperature and high humidity conditions and analyzed using a scanning electron microscope. The microstructure of the membrane was observed and its integrity was scored. The sample group of this invention exhibited a dense and continuous mesh structure with no obvious microcracks or pores; while the membrane surface formed in control group 1 was rough and contained a large number of micro-defects caused by protein aggregation; finally, the membrane was tested on the damaged skin area of ​​the volunteer subjects. test.

[0046] Example 3: This example combines Figures 1 to 3 This describes an ectoine-enhanced recombinant collagen skin barrier repair composition, such as... Figure 1 As shown, the raw material input layer contains 0.5% to 36.0% ectoine as a water molecule structure modifier, deionized water as the balance to construct the solvent environment matrix, and recombinant type III collagen. The recombinant type III collagen exhibits a bimodal distribution, with a ratio of long-chain components with a weight-average molecular weight of 50kDa to 60kDa to short-chain components with a weight-average molecular weight of 3 to 8kDa of 5:1 to 10:1. During the preparation process, ordered water clusters are pre-constructed through step S101, and the mixture is heated to 28 to 32 kDa. Stirring at 450 to 550 rpm for 35 to 45 minutes in an environment induces water molecules to form long-range ordered tetrahedral clusters, thereby obtaining a hydrated matrix with quasi-lattice characteristic physical templates. Step S102 is performed to stretch and anchor low-shear chain segments. Under shear conditions of 60 to 90 rpm and a feeding rate of 0.1 to 0.5 kg / min, the peptide side chains are anchored in the matrix and locked in a linear extended conformation. Finally, a supramolecular hydrated shell with physical steric hindrance and electrostatic shielding effect is formed in the microstructure and functional display layer to repel sodium and chloride ion ablation. The long chain constructs the backbone and the short chain fills the gaps to form a monolayer dense biorepairing network with a thickness of 2 nm to 5 nm. The composition maintains a radius retention rate of ≥95% and a zeta potential fluctuation of <10% in an environment with a mass fraction of 0.9% NaCl.

[0047] like Figure 2 As shown in the figure, the horizontal axis represents the collagen addition rate, which is set between 0.05 and 0.8 kg / min, and the vertical axis represents the specific viscosity of the system. The curves in the figure correspond to the viscosity changes at shear rates of 60 rpm and 90 rpm, respectively. The data shows that within the selected low shear rate range, the steady growth of the system's rheological state can be maintained by controlling the addition rate; Figure 3 As shown, ordered water molecule clusters induced by ectoin molecules on the surface of the stratum corneum construct a dense hydration shell around recombinant collagen through a hydrogen bond network. The figure identifies a repair film layer with a thickness of 2 to 5 nm, formed by long-chain components with a weight-average molecular weight of 50 to 60 kDa and short-chain components with a weight-average molecular weight of 3 to 8 kDa. This film binds to skin tissue through anchoring points in the stratum corneum and utilizes the ion repulsion vector generated by the hydration shell to repel ions from the environment. and Ions, thereby protecting the active conformation of collagen from erosion by environmental electrolytes.

[0048] Example 4: To further clarify the basis for setting key process parameters in the preparation of the ectoin-enhanced recombinant collagen skin barrier repair composition of the present invention, this example systematically explains the principles and calibrates the core parameters in the ordered water cluster preconstruction and low-shear segment stretching steps. In the ordered water cluster preconstruction step, the stirring speed is crucial to whether ectoin can effectively induce water molecules to form long-range ordered tetrahedral clusters. If the speed is too low, the fluid shear force is insufficient to break the random hydrogen bond network of bulk water, making it difficult for ectoin molecules to disperse uniformly and polarize surrounding water molecules. If the speed is too high, the shear stress generated by strong turbulence will destroy the nascent ordered water cluster structure, leading to a decrease in the stability of the hydrated matrix. Therefore, this example designs a set of gradient stirring experiments, setting the speed range as follows: to ,by For gradient, in Stirring under constant temperature conditions By nuclear magnetic resonance hydrogen spectrum Determine the transverse relaxation time of water molecules in the system This characterizes the degree of restriction on the movement of water molecules and their level of order.

[0049] The test results show that as the rotational speed increases from... Upgraded to The system A decrease in the value indicates an increased degree of confinement of water molecules, suggesting the formation of an ordered structure; when the rotational speed is at... to When the interval is, When the value reaches its minimum and remains stable, it indicates that the fluid shear force and intermolecular forces are in optimal balance, forming a stable ectoine-hydrated matrix; however, when the rotational speed exceeds... back, The value begins to rise, indicating that excessive shear force has caused the disintegration of some ordered structures. This invention limits the stirring speed during the ordered water cluster pre-construction step to [specific value]. to In the low-shear chain stretching step, matching the addition rate of recombinant type III collagen with the system's shear rate is crucial to ensuring the correct unfolding and anchoring of the protein peptide chains in the hydrated matrix. If the addition rate is too fast or the shear rate is too low, protein molecules are prone to forming local concentration centers and self-aggregating. If the shear rate is too high, although it facilitates dispersion, it may sever the weak hydrogen bond anchoring sites that are forming between the protein molecules and the hydrated matrix. In this embodiment, the optimal parameter window is determined by monitoring the characteristic viscosity change of the system, and the system's base shear rate is set to [value missing]. to At different mass flow rates to Add recombinant type III collagen.

[0050] Data shows that when the addition speed is controlled at to And the shear rate is maintained at to At that time, the specific viscosity of the system increased linearly and steadily, without any abrupt viscosity changes due to molecular aggregation, and the circular dichroism chromatogram of the final product was also consistent. Analysis showed that the triple helix structure of collagen remained intact. The above applies once the addition speed exceeds [a certain threshold]. The system exhibits fluctuations in non-Newtonian fluid properties, indicating the formation of non-uniform aggregates at the microscopic level. Therefore, a specific matching range between the feeding rate and shear rate in this step is crucial to ensure that the recombinant collagen achieves uniform dispersion and conformational locking at the monomolecular level within the hydrated matrix. In summary, this embodiment reveals, through the principled decomposition and quantitative calibration of key process parameters, the following principles are demonstrated. to Pre-construction speed and to The feeding rate is a precise engineering choice based on the equilibrium of fluid dynamics and molecular thermodynamics, rather than an arbitrary empirical setting.

[0051] Example 5: To ensure that the ectoin-enhanced recombinant collagen skin barrier repair composition of the present invention maintains consistent microstructural characteristics and macroscopic repair efficacy across different production batches, this example establishes a standardized offline calibration and quality control procedure for the interaction strength between the core functional component ectoin and recombinant type III collagen. Quantitative methods are used to monitor the orderliness of the hydration matrix and the locking efficiency of protein conformation, thereby eliminating performance fluctuations that may be introduced by minor differences in raw materials. During the offline calibration stage, the relationship between ectoin concentration and the transverse relaxation time of water molecules in the system is established. The standard correlation curve was prepared at a mass percentage concentration of... to Ectocin aqueous solution, using low-field nuclear magnetic resonance technology in The following measurements The data fitting results show that as the concentration of ectoine increases, The value shows a non-linear decreasing trend, and reaches a certain concentration. to The inflection point within the interval indicates that the efficiency of free water to bound water conversion has reached its peak. Based on this curve, this invention sets the intermediate product of the hydrated matrix in the production process... Acceptance threshold, i.e., only when the actual measurement The value falls within the predicted value of the standard curve. Only when the deviation is within the acceptable range can the batch of hydrated matrix be considered qualified.

[0052] In addition, differential scanning calorimetry is introduced in this embodiment to address the batch stability of recombinant type III collagen. As a key quality control measure for conformation locking, different batches of recombinant type III collagen raw materials were taken and dispersed in a standard ectoine hydrate matrix according to the proportions of this invention, and their thermal denaturation temperature was measured. A qualified conformational locking state should exhibit the following characteristics: The value is at least higher than that of pure collagen aqueous solution. Furthermore, the endothermic peak half-width narrows, which directly reflects the physical stabilizing effect of the ectoin-induced hydration shell on the protein triple helix structure. If a batch of raw materials in a standard matrix... The increase was less than If this is not the case, a pre-calibration procedure needs to be initiated, by fine-tuning the initial concentration of ectoine or the pre-construction stirring time until... The indicators returned to the acceptable range. Furthermore, to address potential differences in shear efficiency of mixing equipment during actual production, this embodiment establishes an on-site commissioning procedure based on characteristic viscosity. Before the low-shear chain segment stretching step of each batch begins, a small sample is mixed, and the rate of increase in the system's specific viscosity is monitored in real time. If the rate of increase deviates from the standard linear model by more than [a certain amount], [further action is taken]. By adjusting the speed of the agitator or the feeding position, the rheological behavior of the system can be brought back to the characteristic range of Newtonian fluid.

[0053] Example 6: To address the inconsistent dissolution behavior of different batches of recombinant type III collagen raw materials due to differences in molecular weight distribution, and to establish an on-site calibration process to ensure the quality of the ectoin hydration matrix in the ordered water cluster preconstruction step, this example establishes the following standardized engineering procedure: for each newly arrived batch of recombinant type III collagen, gel permeation chromatography is used... Determine its weight-average molecular weight and polydispersity coefficient If measured In to The edge of the preferred range, or Exceed If solubility is not achieved, a solubility correction procedure needs to be initiated. This involves adjusting the stirring rate and temperature based on the standard dissolution process to compensate for differences in the raw materials until the intrinsic viscosity of that batch of raw materials in the standard solvent is corrected. The deviation from the reference value is less than .

[0054] Secondly, regarding the construction quality of the ectoin hydration matrix, this embodiment introduces an online relaxation time monitoring procedure based on low-field nuclear magnetic resonance technology. During the ordered water cluster pre-construction step, the transverse relaxation time of water molecules in the system is collected in real time. Data, only when The value exhibits a characteristic rapid decline and then stabilizes at... to The interval lasts for at least When the system determines that the hydrated matrix has constructed an ordered tetrahedral cluster structure that can effectively lock the protein conformation, it automatically triggers the feeding command for the low-shear chain stretching step. If it detects... If the value fluctuates abnormally or fails to reach the preset steady-state range, the stirring speed will be automatically adjusted or the stirring time will be extended through the feedback control algorithm until the microscopic order of the hydrated matrix meets the preset physical index requirements.

[0055] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the present invention.

[0056] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A recombinant collagen skin barrier repair composition with ectoine enhancement, characterized in that, Includes the following components by mass percentage: Reorganization Type II collagen to ; Ikdoin to ; Deionized water is used as a reserve to make up the difference. ; Among them, restructuring The mass ratio of type 3 collagen to ectoine is: to Reorganization Type II collagen consists of collagen with a weight-average molecular weight of to The long-chain components and weight-average molecular weight are to It consists of short-chain components, and the mass ratio of long-chain components to short-chain components is [missing information]. to ; The composition is prepared by the following steps: step Ikdoin in ℃ to Dissolves in deionized water at ℃ and is used as to Stirring speed to A hydrated matrix is ​​obtained; step ,exist to Under shear conditions, recombinant materials are added to the hydrated matrix. Type II collagen, using fluid shear force to reorganize The peptide side chains of type II collagen are anchored in the hydrated matrix to form a linear extended conformation within the hydrated matrix; the composition contains a mass fraction of In a sodium chloride solution environment, recombinant The hydration kinetic radius retention rate of type II collagen is not less than ,and Potential fluctuations are less than .

2. The ectoin-enhanced recombinant collagen skin barrier repair composition according to claim 1, characterized in that: Long-chain components construct a bio-repair framework on the surface of the stratum corneum, while short-chain components are distributed within the gaps of this framework; recombinant Type II collagen forms a hydrated shell in the hydrated matrix with a physical steric hindrance effect. This hydrated shell repels sodium and chloride ions in the sodium chloride solution environment, thus promoting recombination. Type II collagen at an ionic strength of It maintains its active conformation under certain conditions.

3. The ectoin-enhanced recombinant collagen skin barrier repair composition according to claim 1, characterized in that: It also includes trehalose and sodium hyaluronate, which together with ectoine form a synergistic hydration system; the osmotic pressure of the composition is... to Its hydration stability index Satisfy the following formula: ,in, The hydration stability index of the composition; The percentage content of ectoine by mass; For reorganization Percentage of type II collagen by mass; This refers to the percentage content of trehalose by mass. Values to , Values to .

4. The ectoin-enhanced recombinant collagen skin barrier repair composition according to claim 1, characterized in that: Reorganization Type II collagen contains a repeating amino acid sequence in which hydroxyproline sites bind to the hydration matrix via a hydrogen bond network, enabling the composition to form a cross-linked hydration repair film at the skin interface.

5. The ectoin-enhanced recombinant collagen skin barrier repair composition according to claim 1, characterized in that: The purity of ectoine is not less than Its mass fraction is aqueous solution in The conductivity at ℃ is lower than Reorganization The isoelectric point of type II collagen is to .

6. The ectoin-enhanced recombinant collagen skin barrier repair composition according to claim 1, characterized in that: step China joins the restructuring The rate of collagen production is to By coordinating fluid shear force with the dipole moment of ectoin, recombination is guided. The peptide chains of type II collagen complete a linear topological arrangement.

7. The ectoin-enhanced recombinant collagen skin barrier repair composition according to claim 1, characterized in that: The composition at a mass fraction of Under stress from sodium chloride solution, recombination The hydration kinetic radius of type II collagen remains at to Within the range.

8. The ectoin-enhanced recombinant collagen skin barrier repair composition according to claim 1, characterized in that: The composition does not contain nonionic surfactants, and the sum of the mass percentages of all components in the composition is [missing information]. .

9. The ectoin-enhanced recombinant collagen skin barrier repair composition according to claim 1, characterized in that: Reorganization The surface of the peptide chain of type II collagen is bound to three continuous water hydration membranes at polar sites. The interlayer hydrogen bond energy of the water hydration membranes is higher than that of the bulk water.

10. The ectoin-enhanced recombinant collagen skin barrier repair composition according to claim 1, characterized in that: After the composition is applied to the surface of the damaged skin, it forms a thickness of [thickness value missing] from the combined long-chain and short-chain components. to A monolayer membrane.

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