CN109966169B

CN109966169B - Polypeptide composition for external use on skin and application thereof

Info

Publication number: CN109966169B
Application number: CN201910327121.XA
Authority: CN
Inventors: 樊锋凯; 黄文�; 林国华
Original assignee: Change Innovation Wuhan Bio Tech Co ltd
Current assignee: Change Innovation Wuhan Bio Tech Co ltd
Priority date: 2019-04-23
Filing date: 2019-04-23
Publication date: 2021-09-03
Anticipated expiration: 2039-04-23
Also published as: CN109966169A

Abstract

The invention discloses a polypeptide composition for external use on skin and application thereof. The skin external polypeptide composition comprises the following components in parts by mole: 3 to 10 parts carnosine, 3 to 6 parts tripeptide GHK, 2 to 5 parts copper peptide, and 6 to 8 parts palmitoyl tetrapeptide. The invention verifies through repeated experiments that 4 polypeptides are screened from more than 20 polypeptides which effectively act on the skin and are quantitatively combined. Experiments prove that the traditional Chinese medicine composition can relieve inflammatory response after hormone use is stopped, shorten the course of disease and has small recurrence risk for glucocorticoid-dependent symptoms. The skin external application agent containing Chang' e composite soothing polypeptide provided by the invention is screened from more than 20 polypeptides which effectively act on the skin and quantitatively combined. Experiments prove that the traditional Chinese medicine composition can relieve inflammatory response after hormone use is stopped, shorten the course of disease and has small recurrence risk for glucocorticoid-dependent symptoms.

Description

Polypeptide composition for external use on skin and application thereof

Technical Field

The invention belongs to the field of daily-use chemical industry, and particularly relates to a polypeptide composition and application thereof.

Background

Glucocorticoids have the effects of suppressing immune response, resisting allergy, resisting inflammation and the like, can relieve congestion and edema after being externally applied, quickly relieve clinical symptoms such as pruritus, skin damage and the like, and are often used for treating facial acute dermatitis. The patients should be stopped after the disease is relieved. But after partial patients are stopped, the disease condition is recurrent, the medicine is repeatedly used by themselves, and the patient can generate dependence on hormone after a long time; especially, the skin diseases of face and pudendum are caused by the long-term use of stronger hormone preparations in large area, which is easy to cause adverse reactions such as skin atrophy, thinning, telangiectasis, pigment spots, hirsutism, skin aging and the like, namely skin sensitivity. Dermatitis caused by long-term repeated improper external use of a hormone is a dermatitis in which the primary skin lesion disappears after the external use of the glucocorticoid, but inflammatory lesions appear after the external use of the glucocorticoid is stopped, and the symptoms are controlled and gradually worsened by repeated use of the glucocorticoid.

In recent years, the onset of disease has been on the rise year by year, and the disease is difficult to cure. At present, the treatment means for the sensitive symptoms of the skin, particularly the glucocorticoid-dependent skin, is very limited, and the moisturizing treatment or the combined treatment with the external non-hormonal immunomodulator is generally adopted, so that the treatment is long and the process is painful, so that a new effective component with the effect of resisting the glucocorticoid-dependent symptoms needs to be developed.

Disclosure of Invention

Aiming at the defects or improvement requirements of the prior art, the invention provides a polypeptide composition for external use on skin and application thereof, aiming at rapidly relieving symptoms in a short time and effectively reducing and inhibiting rebound by compounding polypeptides which affect different metabolic pathways and have different effects and locally and externally applying the polypeptide composition to the affected part of a patient with glucocorticoid dependence, thereby solving the technical problems of long treatment course and painful process of the existing treatment of the glucocorticoid dependence.

According to one aspect of the present invention, there is provided a polypeptide composition for external use on skin, comprising, in terms of mole parts:

3 to 10 parts carnosine, 3 to 6 parts tripeptide GHK, 2 to 5 parts copper peptide, and 6 to 8 parts palmitoyl tetrapeptide.

Preferably, the composition of the polypeptide for skin external application has a molar content of the palmitoyl tetrapeptide of between 30% and 50%.

Preferably, the carnosine mole fraction of the polypeptide composition for external application to skin is 5 to 8.

Preferably, the tripeptide GHK of the polypeptide composition for external use for skin is 4 to 5 parts by mole.

Preferably, the mole fraction of the copper peptide in the polypeptide composition for skin external application is 3 to 4.

According to another aspect of the present invention, there is provided a use of the polypeptide composition for preparing an external dermal pack preparation for anti-glucocorticoid dependence.

According to another aspect of the present invention, there is provided a pack for external application to the skin, comprising the polypeptide composition according to any one of claims 1 to 5.

Preferably, the external skin coating agent comprises a matrix, and the mass percentage of the polypeptide composition is between 0.1% and 0.15%.

Preferably, the external skin pack is a nano emulsion.

Preferably, the skin external pack is a water-in-oil type nanoemulsion.

In general, compared with the prior art, the above technical solution contemplated by the present invention can achieve the following beneficial effects:

the invention verifies through repeated experiments that 4 polypeptides are screened from more than 20 polypeptides which effectively act on the skin and are quantitatively combined. Experiments prove that the traditional Chinese medicine composition can relieve inflammatory response after hormone use is stopped, shorten the course of disease and has small recurrence risk for glucocorticoid-dependent symptoms.

Drawings

FIG. 1 is a facial skin condition of a patient prior to treatment;

FIG. 2 is the skin condition of a patient at 22 days of treatment;

FIG. 3 is a skin condition of a patient after four weeks of treatment;

FIG. 4 is the skin condition at three weeks of follow-up after completion of patient treatment;

figure 5 is a diagram of the skin condition of a patient after the second treatment session is completed.

The left picture is the photo record of the Chang 'e anti-aging luxury version skin decoding robot under white light, and the right picture is the photo record of the Chang' e anti-aging luxury version skin decoding robot under polarized light. The photo recording under white light is close to visual judgment of naked eyes, and the photo recording under polarized light is sensitive to heme, so that the condition of blood vessel hyperplasia on the surface of skin is effectively analyzed.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.

The invention provides a polypeptide composition for external use on skin, which comprises the following components in parts by mole:

3 to 10 parts carnosine, 3 to 6 parts tripeptide GHK, 2 to 5 parts copper peptide, and 6 to 8 parts palmitoyl tetrapeptide; preferably, the method comprises the following steps:

5 to 8 parts carnosine, 4 to 5 parts tripeptide GHK, 3 to 4 parts copper peptide, and 6 to 8 parts palmitoyl tetrapeptide.

Wherein the molar content of the palmitoyl tetrapeptide is between 30% and 50%.

The carnosine sequence is: beta-Ala-His;

the tripeptide GHK sequence is: GLy-His-Lys

The sequence of the copper peptide is as follows: Gly-His-Lys Cu

The sequence of the palmitoyl tetrapeptide is as follows: Pal-Gly-Gln-Pro-Arg-OH

The polypeptide composition provided by the invention is applied to preparing a skin external application agent with desensitization effect, and is preferably applied to preparing an anti-glucocorticoid-dependent skin external application agent.

At present, a wide variety of polypeptides are found to act on skin as signal peptide, carrier peptide and nerve signal inhibiting peptide, and have the effects of resisting oxidation, inhibiting inflammatory reaction and inducing collagen proliferation. However, for glucocorticoid-dependent diseases, the inflammatory response is persistent and acute, while the skin surface environment changes, leading to the phenomenon of refractory and repetitive verification attacks and skin metabolic disorders. No single polypeptide has been found to have a definite effect on complex glucocorticoid dependence. There is no guiding polypeptide application for glucocorticoid dependence. The inventor verifies through repeated experiments that 4 polypeptides are screened from more than 20 polypeptides which effectively act on the skin and are quantitatively combined. Experiments prove that the traditional Chinese medicine composition can relieve inflammatory response after hormone use is stopped, shorten the course of disease and has small recurrence risk for glucocorticoid-dependent symptoms.

Carnosine has antioxidant effect, and can be used for treating skin with effects of resisting photoaging and scavenging free radicals.

The tripeptides GHK and cupreotide are proven to act as activators and antioxidants of tissue remodeling, and are also a signal peptide that promotes the degradation of a large number of collagen aggregates outside the scar, the synthesis of normal collagen in the skin, the production of elastin, proteoglycans and glycosaminoglycans, the growth rate and migration of different cell types, anti-inflammatory, antioxidant responses.

Palmitoyl tetrapeptide accelerates granulocyte chemotactic protein (GCP-2) expression, and promotes wound recovery. Palmitoyl tetrapeptide-7 is a fragment of immunoglobulin IgG, which has many bioactive functions, particularly immunomodulatory functions. A number of in vivo and in vitro studies have reported the function of palmitoyl tetrapeptide-7. Skin cytokines, especially IL6, are involved in chronic inflammatory reactions and play an important role in the aging process of the skin.

The four polypeptides are mixed according to the proportion, show a comprehensive treatment effect on the glucocorticoid dependence, quickly relieve local inflammation, and have obvious skin repair and low recurrence rate. May be due to modulation of local signaling pathways in the skin in the polypeptide composition, inhibiting validation while altering skin metabolism.

However, further experiments prove that the existence form and administration concentration of different polypeptides affect the experimental effect, carnosine in the four polypeptides has the strongest water solubility, but is easy to oxidize, tripeptides GHK and copper peptide have good water solubility, palmitoyl tetrapeptide is fat-soluble polypeptide, and the composition needs to ensure the drug-loading concentration and protect the polypeptide at the same time. The inventor adopts the nano-emulsion, so that the drug loading concentration is improved. Further adopting water-in-oil type nano-emulsion, carnosine which is easy to oxidize is distributed in a water phase and is wrapped by an oil phase to obtain a better preservation effect; when the nano emulsion selected and prepared in the embodiment of the invention is coated on the surface of skin, a stable oily coating film can be formed in a long time, the transdermal effect of the polypeptides is improved, the transdermal synergistic effect of the four polypeptides is ensured, and the nano emulsion is suitable for being used as a skin external application coating agent.

The invention provides a coating agent for external application to skin, which comprises the polypeptide composition and a nano emulsion matrix.

The polypeptide composition accounts for 0.1 to 0.15 percent by mass.

The nanoemulsion matrix is preferably a water-in-oil nanoemulsion.

The following are examples:

a skin external polypeptide composition comprises the following components in parts by mole:

examples 6 to 10

A skin external pack, in the form of a polypeptide composition according to examples 1 to 5, is formulated as follows:

the preparation method comprises the following steps: adding the polypeptide composition in the embodiments 1 to 5 into a phosphate buffer according to a mixture ratio, adding soybean lecithin, yellow collagen, polyglutamic acid and glycerol, and fully dissolving to obtain a water phase; mixing other components in the ingredient list to obtain an oil phase; mixing the water phase and the oil phase to obtain coarse emulsion, performing ultrasonic treatment at 5 ℃ for 2min with ultrasonic power of 500w, and then performing high-pressure micro-jet homogenization treatment at pressure of 80MPa for three times.

Examples 11 to 15

EXAMPLE 16DPPH radical scavenging Rate

Example 3 and example 5, the results of the DPPH radical scavenging experiments are shown in the table

Experiments show that the clearance rate of the polypeptide compositions in the example 3 and the example 5 is higher than that of the single component, so that the examples 3 and 5 have better DPPH free radical scavenging capacity.

Example 17 inhibition of Hyaluronidase Activity

The results of experiments on the inhibition rate of hyaluronidase activity in examples 3 and 5 are shown in the following table

Experiments show that the clearance rate of the polypeptide compositions in the example 3 and the example 5 is higher than that of the single use of any component, so that the example 3 and the example 5 have better hyaluronidase activity inhibition capability.

EXAMPLE 18 clinical testing

80 patients with facial glucocorticoid dermatitis in the tested population have an average course of 12 months and are randomly divided into 4 groups of 20 persons, and the composition structure is as follows:

a first group: male 3 cases (of which 1 is diffuse type), female 17 cases (of which 2 are diffuse type);

second group: 4 men (1 of them was diffuse), 16 women;

third group: male 3 cases, female 17 cases;

and a fourth group: male 2 cases (1 of them was diffuse type) and female 18 cases.

The test protocol was as follows: the results of the test were observed by applying twice in the morning and evening the coating agent prepared in example 8 and example 15, respectively, a positive control (0.1% tacrolimus cream), and a blank control (the nanoemulsion used in examples 6 to 15 without the polypeptide composition).

Average relief of early stinging test: the relief of the sore pain at the affected area was observed within one week for each group of subjects as shown in the following table:

and (3) effective rate test: before and after 4 weeks of treatment, patients were assessed for score of symptoms including telangiectasia, erythema, desquamation, pustules, papules, skin atrophy, pigmentation, each symptom was classified as "light", "medium" and "heavy", scores were 1, 2 and 3, respectively, and the sum of all symptoms was used as score of symptoms, and the efficacy parameters were calculated as follows:

observing the recovery condition of the affected part in the skin around the periphery, and calculating the curative effect parameters, wherein the curative effect parameters of more than or equal to 90 percent are cured, the curative effect parameters of more than or equal to 60 percent are effective, the curative effect parameters of more than or equal to 20 percent are effective, and the curative effect parameters of less than 20 percent are ineffective. The effective rate test results are as follows:

the effective rate test results show that the total effective rates of the external skin application agents provided in the examples 8 and 15 are similar to the positive control tacrolimus cream and are obviously higher than the blank control; in terms of cure rate, example 8 (40%) was similar to the skin external pack provided in example 15 (45%), and was significantly higher than the positive control (20%) and the blank control (15%).

Onset time test: the effect of daily medication is observed, the efficacy parameters are calculated, and the onset time is determined when 14 patients with efficacy parameters exceeding 20% appear in each group. Onset time test results are given in the following table:

the results of the onset time test show that the external dermal application provided in examples 8 and 15 have shorter onset time than the positive control, and may stimulate the inhibition of hyaluronidase and the stimulation of collagen proliferation by the polypeptide composition, so the effect on the symptoms of glucocorticoid-dependent patients is more obvious than that of the tacrolimus cream with single action. On the other hand, the external dermal liniment provided in example 8 was superior in the overall effectiveness to the external dermal liniment provided in example 15, and was slightly longer in the effect-developing time than the external dermal liniment provided in example 15, and the difference was not significant.

And (3) observation of a rebound phenomenon: after four weeks of the first to third groups of medicines, the medicines are stopped, the moisturizing skin care product is coated in the morning and evening, and the rebound phenomenon is observed after follow-up after three weeks. The results are as follows:

all 60 patients were followed without one missed visit. Wherein: only 1 of the first group of patients showed rebound (diffuse type patients); patients in the second group who did not exhibit rebound; patients in the third group presented with 3 rebound phenomena. The result shows that the skin external polypeptide composition provided by the invention can effectively avoid the rebound phenomenon of glucocorticoid dependence and has obvious desensitization effect.

Among the first group of patients, patients who developed the rebound phenomenon were recorded on their facial skin conditions as shown in fig. 1 to 5 using a skin analyzer (Chang ' e anti-aging luxury version skin decoding robot) developed by Chang ' e medical anti-aging robot incorporated in Wuhan's medical service before the treatment with the skin external pack provided in example 8, at the time of onset, after the treatment was completed all around, and at the time of follow-up. FIG. 1 shows that the patient is a diffuse glucocorticoid dependent patient, male; figure 2 is a skin analysis record three weeks after treatment showing onset of action; FIG. 3 shows a significant improvement in skin after four weeks of treatment, but with symptoms of telangiectasia, desquamation, and skin atrophy; FIG. 4 shows a certain degree of rebound after the administration, which is still significantly relieved compared with the original condition. One week after the follow-up visit, the second course of treatment was performed, and after continuously using the external application agent provided in example 8 for 4 weeks as shown in fig. 5, the healing was achieved without the occurrence of rebound phenomenon.

The skin external pack prepared in example 8 has been developed as "Chang' e composite soothing polypeptide" product.

It will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.

Claims

1. The use of a polypeptide composition for preparing a skin external pack preparation for treating glucocorticoid dependence; the polypeptide composition comprises the following components in parts by mole:

2. Use of a polypeptide composition according to claim 1, wherein the palmitoyl tetrapeptide is present in a molar amount of between 30% and 50%.

3. Use of a polypeptide composition as claimed in claim 1 wherein the carnosine is present in a molar fraction of 5 to 8 parts.

4. Use of a polypeptide composition according to claim 1 wherein the tripeptide GHK is present in a molar fraction of 4 to 5 parts.

5. The use of the polypeptide composition of claim 1, wherein the copper peptide is present in a molar amount of 3 to 4 parts.

6. The use of the polypeptide composition of claim 1, wherein the topical skin coating comprises a base material, and the polypeptide composition is present in an amount of between 0.1% and 0.15% by weight.

7. The use of the polypeptide composition of claim 6, wherein the skin external application coating agent is a nanoemulsion.

8. Use of a polypeptide composition according to claim 7, wherein the nanoemulsion is a water-in-oil nanoemulsion.