CN113476593A - Application of recombinant humanized collagen, and related composition and preparation method thereof - Google Patents
Application of recombinant humanized collagen, and related composition and preparation method thereof Download PDFInfo
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Abstract
The invention relates to application of recombinant humanized collagen, and a related composition and a preparation method thereof. Specifically, the invention provides application of the recombinant humanized collagen in preparing a product for preventing and/or treating allergic skin diseases (particularly eczema), and also provides a composition containing the recombinant humanized collagen and a preparation method thereof. Experiments prove that the recombinant humanized collagen can effectively relieve the skin red swelling, erythema, edema, pruritus and inflammatory reaction caused by eczema, and can restore the skin luster, and the composition containing the recombinant humanized collagen can more quickly relieve the eczema symptom compared with a simple recombinant humanized collagen aqueous solution.
Description
Technical Field
The invention belongs to the field of biomedicine, and relates to application of recombinant humanized collagen, a related composition of the recombinant humanized collagen and a preparation method of the composition.
Background
Infantile eczema is a common disease of children, and children are not easy to administer the medicine due to low autoimmunity and weak liver and kidney functions. The common products such as skin cream, skin cream and the like contain a small amount of hormone, and can cause adverse effects on children after long-term use. According to clinical reports, after products such as skin cream, skin cream and the like are used for a long time, the skin of children can generate drug resistance, so that the effect is lower and lower. Therefore, there is a need to develop more hormone-free products for treating skin eczema, providing more options for patients.
The patent application with application No. 201811438582.6 entitled "polypeptide, method for producing the same, and use thereof" and the patent application with application No. 201210482543.2 entitled "a recombinant human collagen and method for producing the same" describe recombinant humanized collagen that the inventors of the present invention studied and produced before, but do not directly disclose its specific use in the biomedical field.
Disclosure of Invention
Problems to be solved by the invention
The present invention intends to use the recombinant humanized collagen described in application nos. 201811438582.6 and 201210482543.2 as a raw material to evaluate its effectiveness for preventing and/or treating allergic skin diseases (particularly eczema), and to provide a composition containing the recombinant humanized collagen for preventing and/or treating related diseases, and a method for preparing the composition.
Means for solving the problems
In a first aspect, the invention provides the use of a recombinant humanized collagen comprising n repeats of the sequence shown as SEQ ID No.1, wherein n is an integer greater than or equal to 1, in the manufacture of a product for the prevention and/or treatment of allergic skin diseases; when n is an integer of 2 or more, the repeated sequences are directly linked or separated by one or more amino acid residues.
Further, in the above use, the recombinant humanized collagen optionally comprises a sequence shown as SEQ ID No.2, wherein when the recombinant humanized collagen comprises a sequence shown as SEQ ID No.2, the n repeats of the sequence shown as SEQ ID No.1 and the sequence shown as SEQ ID No.2 are directly linked or separated by one or more amino acid residues.
Preferably, in the above use, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 20, 24 or 32, wherein when n is an integer of 2 or more, the repeated sequences are directly linked or are separated by one or more amino acid residues.
Further, in the above use, the allergic skin disease is eczema.
Preferably, in the above use, the eczema is infantile eczema.
Further, in the above use, the eczema has at least one of the following symptoms: redness and swelling of the skin, erythema of the skin, edema of the skin, lack of radiance of the skin, itching of the skin and elevated expression of inflammatory immune factors.
Preferably, in the above use, the inflammatory immune factor comprises IL-4.
In a second aspect, the present invention provides a composition comprising an a phase, a B phase, a C phase, and a D phase, wherein the a phase comprises, in mass percent: 1.1-5.5% of emulsifier, 10-12% of softening agent, 2-3% of thickening agent and 2.5-3% of sun-screening oil; the phase B comprises: 0.13 to 0.75 percent of preservative, 0.3 to 0.5 percent of rheology regulator, 2 to 3 percent of active ingredient protective agent, 3 to 7 percent of humectant and the balance of water; the phase C comprises: 0.1 to 0.2 percent of moisturizer and 3 to 4 percent of antibacterial agent; the phase D comprises: 0.05 to 0.15 percent of recombinant humanized collagen; the recombinant humanized collagen comprises n repeats of a sequence shown by SEQ ID No.1 and an optional sequence shown by SEQ ID No.2, wherein n is an integer more than or equal to 1; when n is an integer of 2 or more, the repeated sequences are directly connected or separated by one or more amino acid residues; when the recombinant humanized collagen comprises a sequence shown as SEQ ID No.2, the n repeats of the sequence shown as SEQ ID No.1 and the sequence shown as SEQ ID No.2 are directly connected or separated by one or more amino acid residues.
Further, in the above composition, the emulsifier comprises at least one of M68 emulsifier, a165 emulsifier, and cetearyl alcohol; the emollient comprises liquid paraffin; the thickener comprises polydimethylsiloxane; the sunscreen oil comprises olive fruit oil; the preservative comprises at least one of methylparaben, propylparaben and phenoxyethanol; the rheology modifier comprises carbomer; the active ingredient protectant comprises trehalose; the humectant comprises glycerin; the moisturizing agent comprises triethanolamine; the antimicrobial agent comprises aloe vera extract.
In a third aspect, the present invention provides a process for preparing the above composition, comprising the steps of: s1: respectively heating the phase A component and the phase B component in the prescription amount under the stirring condition and uniformly mixing; s2: adding the phase A component into the phase B component, preserving heat, stirring, and cooling to obtain a first mixture; s3: adding the C phase component with the prescription amount into the first mixture, and homogenizing to obtain a second mixture; s4: adding the D phase component with the prescription amount into the second mixture, and homogenizing until the system is uniform, thus obtaining the composition.
In a fourth aspect, the present invention provides a product comprising the above composition and optionally pharmaceutically or cosmetically acceptable excipients.
ADVANTAGEOUS EFFECTS OF INVENTION
Through the implementation of the technical scheme, the recombinant humanized collagen can relieve the symptoms of skin redness and swelling, erythema, edema, pruritus and the like caused by eczema and reduce the inflammatory reaction caused by the eczema, and the composition containing the recombinant humanized collagen can further accelerate the relief of the symptoms of the eczema and promote the skin to recover luster.
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FIG. 1 shows the results of measurement of IL-4 expression levels at different periods in rats in each group.
Detailed Description
The following describes embodiments of the present invention, but the present invention is not limited to these embodiments. The present invention is not limited to the configurations described below, and various modifications are possible within the scope of the claims, and embodiments and examples obtained by appropriately combining the technical means disclosed in the respective embodiments and examples are also included in the technical scope of the present invention.
Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
In the present specification, a numerical range represented by "a value to B value" or "a value to B value" means a range including the end point value A, B.
In the present specification, the meaning of "may" includes both the meaning of performing a certain process and the meaning of not performing a certain process. In this specification, "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event occurs and instances where it does not.
In the present specification, "%" indicates a mass percentage, i.e., g/100g, unless otherwise specified.
Reference throughout this specification to "some particular/preferred embodiments," "other particular/preferred embodiments," "some particular/preferred aspects," "other particular/preferred aspects," or the like, means that a particular element (e.g., feature, structure, property, and/or characteristic) described in connection with the embodiment is included in at least one embodiment described herein, and may or may not be present in other embodiments. In addition, it is to be understood that the described elements may be combined in any suitable manner in the various embodiments.
The recombinant humanized collagen of the present invention is described in patent application No. 201811438582.6 entitled "polypeptide, its production method and use" and patent application No. 201210482543.2 entitled "a recombinant humanized collagen and its production method". Among them, the recombinant humanized collagen described in the patent application No. 201811438582.6 was used in COL1 group in examples, and the recombinant humanized collagen described in the patent application No. 201210482543.2 was used in COL2 group in examples.
In one embodiment, the recombinant humanized collagen of the present invention comprises N repeats of the sequence shown in SEQ ID No.1 (consisting of 30 amino acids, GERGAPGFRGPAGPNGIPGEKGPAGERGAP in order from N-terminus to C-terminus), wherein N is an integer of 1 or more, and when N is an integer of 2 or more, the repeats are directly linked or separated by one or more amino acid residues.
In a preferred embodiment, the recombinant humanized collagen of the present invention comprises n repeats of the sequence shown as SEQ ID No.1, wherein n is an integer of 1 or more, and when n is an integer of 2 or more, the repeats are directly linked.
In a more preferred embodiment, the recombinant humanized collagen of the present invention comprises n repeats of the sequence shown as SEQ ID No.1, n being 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 20, 24 or 32, preferably 16, wherein when n is an integer of 2 or more, the repeats are directly linked.
In a more preferred embodiment, the recombinant humanized collagen of the present invention comprises 16 repeats of the sequence shown in SEQ ID No.1, and the repeats are directly linked to each other, i.e. the sequence shown in SEQ ID No.3 (480 aa in total length, GERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAP in sequence from N-terminus to C-terminus); the recombinant humanized collagen used in COL1 group in the examples of the present invention contained the sequence shown in SEQ ID No. 3.
In one embodiment, the recombinant humanized collagen of the present invention comprises N repeats of the sequence shown as SEQ ID No.1 and a sequence shown as SEQ ID No.2 (consisting of 10 amino acids, GPPGPCCGGG from N to C), wherein N is an integer of 1 or more, and when N is an integer of 2 or more, the repeats are directly linked or separated by one or more amino acid residues, and the N repeats of the sequence shown as SEQ ID No.1 and the sequence shown as SEQ ID No.2 are directly linked or separated by one or more amino acid residues.
In a preferred embodiment, the recombinant humanized collagen of the present invention comprises n repeats of the sequence shown as SEQ ID No.1 and the sequence shown as SEQ ID No.2, wherein n is an integer of 1 or more, and when n is an integer of 2 or more, the repeats are directly linked or separated by one or more amino acid residues, and the n repeats of the sequence shown as SEQ ID No.1 and the sequence shown as SEQ ID No.2 are separated by one or more amino acid residues.
In a more preferred embodiment, the recombinant humanized collagen of the present invention comprises n repeats of the sequence shown as SEQ ID No.1 and the sequence shown as SEQ ID No.2, n being 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 20, 24 or 32, preferably 16, wherein when n is an integer equal to or greater than 2, the repeats are directly linked or are separated by one or more amino acid residues; the n repeats of the sequence shown as SEQ ID No.1 and the sequence shown as SEQ ID No.2 are separated by one or more amino acid residues.
In a more preferred embodiment, the recombinant humanized collagen of the present invention comprises 16 repeats of the sequence shown as SEQ ID No.1 and the sequence shown as SEQ ID No.2, each repeat being directly linked or separated by 2 amino acid residues, and the 16 repeats of the sequence shown as SEQ ID No.1 and the sequence shown as SEQ ID No.2 being separated by 5 amino acid residues, i.e. the sequence shown as SEQ ID No.4 (full length 501aa, GERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPRSGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPRSGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPRSGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPGERGAPGFRGPAGPNGIPGEKGPAGERGAPRSPEFGPPGPCCGGG from N-terminus to C-terminus); the recombinant humanized collagen used in COL2 group in the examples of the present invention contained the sequence shown in SEQ ID No. 4.
In one embodiment, the allergic skin disease of the present invention is eczema.
In a preferred embodiment, the allergic skin disease according to the invention is infantile eczema.
In one embodiment, the composition comprising recombinant humanized collagen of the present invention comprises a phase a, a phase B, a phase C and a phase D, wherein the phase a comprises, in mass percent: 1.1-5.5% of emulsifier, 10-12% of softening agent, 2-3% of thickening agent and 2.5-3% of sun-screening oil; the phase B comprises: 0.13 to 0.75 percent of preservative, 0.3 to 0.5 percent of rheology regulator, 2 to 3 percent of active ingredient protective agent, 3 to 7 percent of humectant and the balance of water; the phase C comprises: 0.1 to 0.2 percent of moisturizer and 3 to 4 percent of antibacterial agent; the phase D comprises: 0.05 to 0.15 percent of recombinant humanized collagen; the emulsifier comprises at least one of an M68 emulsifier, an A165 emulsifier, and cetearyl alcohol; the emollient comprises liquid paraffin; the thickener comprises polydimethylsiloxane; the sunscreen oil comprises olive fruit oil; the preservative comprises at least one of methylparaben, propylparaben and phenoxyethanol; the rheology modifier comprises carbomer; the active ingredient protectant comprises trehalose; the humectant comprises glycerin; the moisturizing agent comprises triethanolamine; the antimicrobial agent comprises aloe vera extract.
Unless otherwise indicated, instruments, reagents, materials, laboratory animals and the like used in the present invention are commercially available in a conventional manner.
Example 1: recombinant humanized collagen for treating eczema
1. Recombinant humanized collagen relieves skin symptoms caused by eczema
1.1 establishment of Experimental animal models (refer to the Standard for preparation of eczema animal models (draft))
42 rats were divided into seven groups, i.e., a control group, a COL1 low-concentration group, a COL1 medium-concentration group, a COL1 high-concentration group, a COL2 low-concentration group, a COL2 medium-concentration group, and a COL2 high-concentration group, each of which was 6 rats. The skin of the rat back was divided into A, B parts, where part A was near the neck and part B was the back, and depilated for part A, approximately 2cm by 2cm, and part B approximately 4cm by 4 cm. A7% acetone solution of 2, 4-Dinitrochlorobenzene (DNCB) was prepared, and 100. mu.L of the acetone solution was pipetted and applied to the site A to cause sensitization.
After 1 week, 200. mu.L of DNCB acetone solution was applied to B for excitation. And then exciting once every 5d, and after 6 times of total excitation, successfully establishing the model.
As a result: the animals of group 4 were given DNCB acetone solution at A and the rats developed severe itching movements, rolling frequently, agitation. After 6 times of excitation, erythema, edema and scratch (skin scratch) appear on the skin at the position B, and the skin is rolled violently and the itching action is obvious. Therefore, the animal model can be successfully established.
1.2 Experimental methods
After the model is established, the normal saline is given to the control group every day, and the normal saline solution containing the recombinant humanized collagen is respectively given to the COL1, the COL2 low-concentration group, the COL1 and the COL2 medium-concentration group, the COL1 and the COL2 high-concentration group at 0.5mg/mL, 1mg/mL and 3mg/mL (the normal saline solution containing the recombinant humanized collagen is self-prepared, and is formed by adding the normal saline into the recombinant humanized collagen freeze-dried powder self-produced by the inventor group for dissolving). 1 time a day, 1mL each time. When the medicine is administrated, a pipettor is required to slowly drip 200 mu L of the medicine for each drip. For better absorption by the rat skin. The administration was for a total of 9 days, and the skin condition, itching condition, and skin gloss on the back of the rats were recorded.
1.3 results of the experiment
Specific rat conditions were recorded as follows:
from the experimental results, it can be known that the administration of the physiological saline solution containing the recombinant humanized collagen can significantly improve the red swelling, erythema and pruritus at the stimulated part of the rat, and can accelerate the relief of various symptoms caused by eczema. The two recombinant humanized collagen proteins in the COL1 and COL2 groups are relatively close in the effect of treating eczema.
2. Recombinant humanized collagen reduces inflammatory reaction generated by eczema
2.1 establishment of experimental animal models (refer to the Standard for preparation of eczema animal models (draft))
60 rats are taken for model building, and the specific operation is 1.1.
2.2 Experimental methods
After the model is established, the normal saline is given to the control group every day, and the normal saline solution containing the recombinant humanized collagen is respectively given to the COL1 group, the COL2 low-concentration group, the COL1 group, the COL2 medium-concentration group, the COL1 group and the COL2 high-concentration group at 0.5mg/mL, 1mg/mL and 3mg/mL (the normal saline solution containing the recombinant humanized collagen is self-prepared, and is formed by adding the normal saline into the recombinant humanized collagen freeze-dried powder self-produced by the inventor group for dissolving). 1 time a day, 1mL each time. When the medicine is administrated, a pipettor is required to slowly drip 200 mu L of the medicine for each drip. For better absorption by the rat skin.
Of these, 4 rats were sacrificed prior to the first day of administration, and B subcutaneous tissues were taken and examined for IL-4. The remaining 56 were divided into seven groups for administration and feeding.
On the fifth day, 4 of the control group, COL1, COL2 low concentration group, COL1, COL2 medium concentration group, COL1, and COL2 high concentration group were sacrificed. And taking subcutaneous tissues at B, and detecting IL-4.
On the ninth day, the remaining control group, COL1, COL2 low concentration group, COL1, COL2 medium concentration group, COL1, COL2 high concentration group, 4 each, were sacrificed. And taking subcutaneous tissues at B, and detecting IL-4.
Then, the relative IL-4 value of each part of subcutaneous tissue is detected by a fluorescent quantitative PCR detection method.
2.3 results of the experiment
The results of the experiment are shown in fig. 1, the relative value of IL-4 is highest in the blank model sacrificed on the day of the experiment, and then at 5d, it is clearly seen that the value of IL-4 is significantly reduced when rats containing the physiological saline solution of the recombinant humanized collagen are given, whereas at 9d, the value of IL-4 is more reduced when rats containing the physiological saline solution of the recombinant humanized collagen are given relative to the control group, which proves that the physiological saline solution containing the recombinant humanized collagen can significantly reduce the inflammatory response caused by eczema and is in positive correlation with the concentration. The results of two recombinant humanized collagen proteins in the COL1 and COL2 groups were closer in reducing the expression of IL-4.
3. Related research of composition containing recombinant humanized collagen
3.1 preparation of the formula (in mass percent)
3.2 preparation method
(1) Adding the raw material of the component A into an oil phase pot, stirring and heating to 80 +/-2 ℃;
(2) heating the raw materials of the component B to 80 +/-2 ℃ in a water phase pot, and uniformly mixing;
(3) adding the component A into the component B, stirring, homogenizing for several minutes, keeping the temperature at 80 +/-2 ℃, stirring for 15min, and cooling;
(4) adding the raw material of the component C when the temperature T is 40 ℃, homogenizing for a plurality of minutes, and measuring the pH value;
(5) adding the component D (0.22 μm filter membrane for filtration), and homogenizing for several minutes to make the system uniform;
(6) and (4) inspecting the semi-finished product, filling and packaging after the semi-finished product is inspected to be qualified, inspecting the finished product, and warehousing after the finished product is inspected to be qualified.
3.3 pharmacodynamic experiments
And (3) taking 18 Wistar rats for model establishment, and performing the specific operation as 1.1.
After the animal model was established, the rats were divided into 3 groups, i.e., a control group to which 1mL of physiological saline was administered daily, a COL1 administration group to which 1g of the above composition containing about 1mg of recombinant humanized collagen was applied daily, and a COL2 administration group to which skin conditions of the rats were recorded.
The specific results are shown in the following table, and it can be seen from the table that after the composition containing the recombinant humanized collagen is applied, the degree of redness and edema at the B site can be obviously reduced, and the recovery speed is faster than that of the composition containing the recombinant humanized collagen in the above step 1. In addition, the two recombinant humanized collagen proteins in the COL1 and COL2 groups have almost the same effect after being prepared into preparations.
Sequence listing
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100 105 110
Pro Ala Gly Glu Arg Gly Ala Pro Arg Ser Gly Glu Arg Gly Ala Pro
115 120 125
Gly Phe Arg Gly Pro Ala Gly Pro Asn Gly Ile Pro Gly Glu Lys Gly
130 135 140
Pro Ala Gly Glu Arg Gly Ala Pro Gly Glu Arg Gly Ala Pro Gly Phe
145 150 155 160
Arg Gly Pro Ala Gly Pro Asn Gly Ile Pro Gly Glu Lys Gly Pro Ala
165 170 175
Gly Glu Arg Gly Ala Pro Gly Glu Arg Gly Ala Pro Gly Phe Arg Gly
180 185 190
Pro Ala Gly Pro Asn Gly Ile Pro Gly Glu Lys Gly Pro Ala Gly Glu
195 200 205
Arg Gly Ala Pro Gly Glu Arg Gly Ala Pro Gly Phe Arg Gly Pro Ala
210 215 220
Gly Pro Asn Gly Ile Pro Gly Glu Lys Gly Pro Ala Gly Glu Arg Gly
225 230 235 240
Ala Pro Arg Ser Gly Glu Arg Gly Ala Pro Gly Phe Arg Gly Pro Ala
245 250 255
Gly Pro Asn Gly Ile Pro Gly Glu Lys Gly Pro Ala Gly Glu Arg Gly
260 265 270
Ala Pro Gly Glu Arg Gly Ala Pro Gly Phe Arg Gly Pro Ala Gly Pro
275 280 285
Asn Gly Ile Pro Gly Glu Lys Gly Pro Ala Gly Glu Arg Gly Ala Pro
290 295 300
Gly Glu Arg Gly Ala Pro Gly Phe Arg Gly Pro Ala Gly Pro Asn Gly
305 310 315 320
Ile Pro Gly Glu Lys Gly Pro Ala Gly Glu Arg Gly Ala Pro Gly Glu
325 330 335
Arg Gly Ala Pro Gly Phe Arg Gly Pro Ala Gly Pro Asn Gly Ile Pro
340 345 350
Gly Glu Lys Gly Pro Ala Gly Glu Arg Gly Ala Pro Arg Ser Gly Glu
355 360 365
Arg Gly Ala Pro Gly Phe Arg Gly Pro Ala Gly Pro Asn Gly Ile Pro
370 375 380
Gly Glu Lys Gly Pro Ala Gly Glu Arg Gly Ala Pro Gly Glu Arg Gly
385 390 395 400
Ala Pro Gly Phe Arg Gly Pro Ala Gly Pro Asn Gly Ile Pro Gly Glu
405 410 415
Lys Gly Pro Ala Gly Glu Arg Gly Ala Pro Gly Glu Arg Gly Ala Pro
420 425 430
Gly Phe Arg Gly Pro Ala Gly Pro Asn Gly Ile Pro Gly Glu Lys Gly
435 440 445
Pro Ala Gly Glu Arg Gly Ala Pro Gly Glu Arg Gly Ala Pro Gly Phe
450 455 460
Arg Gly Pro Ala Gly Pro Asn Gly Ile Pro Gly Glu Lys Gly Pro Ala
465 470 475 480
Gly Glu Arg Gly Ala Pro Arg Ser Pro Glu Phe Gly Pro Pro Gly Pro
485 490 495
Cys Cys Gly Gly Gly
500
Claims (10)
1. Use of a recombinant humanized collagen comprising n repeats of a sequence shown as SEQ ID No.1, wherein n is an integer greater than or equal to 1, in the manufacture of a product for the prevention and/or treatment of allergic skin diseases; when n is an integer of 2 or more, the repeated sequences are directly linked or separated by one or more amino acid residues.
2. Use according to claim 1, wherein the recombinant humanized collagen optionally comprises the sequence shown as SEQ ID No.2, wherein when the recombinant humanized collagen comprises the sequence shown as SEQ ID No.2, the n repeats of the sequence shown as SEQ ID No.1 and the sequence shown as SEQ ID No.2 are directly linked or separated by one or more amino acid residues.
3. The use according to claim 1 or 2, wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 20, 24 or 32, wherein when n is an integer of 2 or more, the repeated sequences are directly linked or separated by one or more amino acid residues.
4. Use according to any one of claims 1 to 3, characterized in that the allergic skin disease is eczema.
5. Use according to claim 4, characterized in that the eczema has at least one of the following symptoms: redness and swelling of the skin, erythema of the skin, edema of the skin, lack of radiance of the skin, itching of the skin and elevated expression of inflammatory immune factors.
6. The use according to claim 5, wherein said inflammatory immune factor comprises IL-4.
7. A composition comprising an A phase, a B phase, a C phase and a D phase, wherein, in mass percent,
the phase A comprises: 1.1-5.5% of emulsifier, 10-12% of softening agent, 2-3% of thickening agent and 2.5-3% of sun-screening oil;
the phase B comprises: 0.13 to 0.75 percent of preservative, 0.3 to 0.5 percent of rheology regulator, 2 to 3 percent of active ingredient protective agent, 3 to 7 percent of humectant and the balance of water;
the phase C comprises: 0.1 to 0.2 percent of moisturizer and 3 to 4 percent of antibacterial agent;
the phase D comprises: 0.05 to 0.15 percent of recombinant humanized collagen;
the recombinant humanized collagen comprises n repeats of a sequence shown by SEQ ID No.1 and an optional sequence shown by SEQ ID No.2, wherein n is an integer more than or equal to 1; when n is an integer of 2 or more, the repeated sequences are directly connected or separated by one or more amino acid residues; when the recombinant humanized collagen comprises a sequence shown as SEQ ID No.2, the n repeats of the sequence shown as SEQ ID No.1 and the sequence shown as SEQ ID No.2 are directly connected or separated by one or more amino acid residues.
8. The composition of claim 7,
the emulsifier comprises at least one of an M68 emulsifier, an A165 emulsifier, and cetearyl alcohol;
the emollient comprises liquid paraffin;
the thickener comprises polydimethylsiloxane;
the sunscreen oil comprises olive fruit oil;
the preservative comprises at least one of methylparaben, propylparaben and phenoxyethanol;
the rheology modifier comprises carbomer;
the active ingredient protectant comprises trehalose;
the humectant comprises glycerin;
the moisturizing agent comprises triethanolamine;
the antimicrobial agent comprises aloe vera extract.
9. A process for the preparation of a composition according to claim 7 or 8, comprising the steps of:
s1: respectively heating the phase A component and the phase B component in the prescription amount under the stirring condition and uniformly mixing;
s2: adding the phase A component into the phase B component, preserving heat, stirring, and cooling to obtain a first mixture;
s3: adding the C phase component with the prescription amount into the first mixture, and homogenizing to obtain a second mixture;
s4: adding the D phase component with the prescription amount into the second mixture, and homogenizing until the system is uniform, thus obtaining the composition.
10. A product comprising a composition according to claim 7 or 8 and optionally a pharmaceutically or cosmetically acceptable adjuvant.
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| CN115487348A (en) * | 2022-09-23 | 2022-12-20 | 广东粤港澳大湾区黄埔材料研究院 | Composition containing recombinant humanized collagen and preparation method and application thereof |
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