External dressing based on phase transition lysozyme loaded biguanide drugs and preparation method of external dressing

Abstract

The invention relates to the technical field of medicine preparation, in particular to external dressing based on phase transformation lysozyme loaded biguanide medicine and a preparation method of the external dressing. Co-culturing a proper amount of Kangpu tea strains and the tea sugar water; standing at room temperature for 10-15 days, and culturing a mycoderm with the thickness of 3 mm; filtering the mycoderm with a gauze, fishing out the mycoderm, washing the mycoderm with deionized water for 3-5 times, and centrifuging the mycoderm at the speed of 3000r / min and the temperature of 4 DEG The mycoderm is placed in a 0.1 M NaOH solution with the molecular weight of 40 to be heated, the heating temperature is 90 DEG C, and the heating time is 1 hour; placing the mycoderm in ionized water, and washing the mycoderm in a stirring state; the external dressing has the advantages that the external dressing based on the phase transformation lysozyme loaded biguanide medicine has good transdermal absorption capacity, a protective film can be formed on the skin, the affected part is kept moist, and the effective action time of the medicine is prolonged; in addition, the external dressing based on the phase transition lysozyme loaded biguanide drugs is generally good in tolerance and rare in adverse reaction.

Description

Technical Field

[0001] The invention relates to the technical field of drug preparation, and in particular to an external dressing based on phase transition lysozyme loaded with biguanide drugs and a preparation method thereof. Background Art

[0002] Traditional topical medications for skin diseases mainly include ointments, creams, tinctures, powders, etc. These drugs have relatively simple dosage forms, poor transdermal absorption, short residence time on the skin, and are easily ineffective. In addition, traditional topical medications for skin diseases also have problems such as high irritation and prone to allergies.

[0003] In view of the above problems, a phase-shift lysozyme-loaded biguanide drug topical dressing and a preparation method thereof are proposed. Summary of the invention

[0004] In order to solve the problems raised in the above background technology, the present invention provides a biguanide drug-loaded external dressing based on phase transition lysozyme and a preparation method thereof.

[0005] The technical solution provided by the present invention is: a biguanide drug-loaded external dressing based on phase transition lysozyme and a preparation method thereof, characterized in that it comprises the following steps:

[0006] S1. Co-cultivate an appropriate amount of kombucha bacteria with tea sugar water;

[0007] S2, standing at room temperature to cultivate the biofilm;

[0008] S3. Filter the bacterial membrane with a gauze, rinse with deionized water 3 to 5 times, and centrifuge at 3000 r / min and 4 degrees Celsius for 20 minutes;

[0009] S4, placing the bacterial film in a 0.1M NaOH solution with a molecular weight of 40 and heating it;

[0010] S5. Place the biofilm in deionized water and rinse it clean under stirring;

[0011] S6. Detect the pH value of the biofilm and use filter paper to absorb the moisture of the biofilm;

[0012] S7, low temperature preservation of biofilm;

[0013] S8, use 10mM HEPES buffer as solvent;

[0014] S9, prepare 2 mg / ml Lyz solution A;

[0015] S10, prepare 50mM TCEP (molecular weight 250) solution B (5M NaOH is used to adjust the pH of TCEP to 6.2);

[0016] S11, take 24.172 mg of phenformin drug powder or 12.916 mg of metformin drug powder and mix with 1 ml of solution A, and shake thoroughly to mix;

[0017] S12, add 1 ml of solution B and shake to mix;

[0018] S13. In this mixed system, the proportion of biguanide drugs is 50 mM;

[0019] S14, placing the bacterial film in the above mixed system to obtain an external dressing based on phase-shift lysozyme loaded with biguanide drugs.

[0020] Preferably, the standing time in S2 is 10-15 days, and the thickness of the cultured bacterial film is 3 mm.

[0021] Preferably, the heating temperature in S4 is 90° C. and the heating time is 1 hour.

[0022] Preferably, the volume ratio of the lysozyme liquid to the TCEP liquid in S13 is 1:1, and the proportion of biguanide drugs in the final system is 50 mM.

[0023] The beneficial effects of the present invention are as follows: the topical dressing based on phase-shift lysozyme loaded with biguanide drugs has good transdermal absorption capacity, can form a protective film on the skin, keep the affected area moist, and prolong the effective action time of the drug; in addition, the topical dressing based on phase-shift lysozyme loaded with biguanide drugs is generally well tolerated and has few adverse reactions. DETAILED DESCRIPTION

[0024] The present invention is described below with specific embodiments, but is not intended to be limiting of the present invention.

[0025] The phase-shift lysozyme-loaded biguanide drug external dressing and its preparation method are characterized by comprising the following steps:

[0026] Co-cultivate an appropriate amount of kombucha bacteria with tea sugar water; let it stand at room temperature to cultivate a bacterial film for 10-15 days, and the thickness of the cultured bacterial film is 3mm; filter the bacterial film with a gauze, rinse it with deionized water 3-5 times, and centrifuge it at 3000r / min and 4 degrees Celsius for 20min; place the bacterial film in 0.1M NaOH, a molecular weight of 40 solution and heat it at 90℃ for 1 hour; place the bacterial film in deionized water and rinse it clean under stirring; detect the pH value of the bacterial film, and use filter paper to absorb the moisture of the bacterial film; store the bacterial film at low temperature; use 10mM HEPES buffer as a solvent; prepare 2mg / ml Lyz solution A; prepare 50mM TCEP (molecular weight 250) solution B, and adjust its final pH value to 6.2 with 5M NaOH; take 24.172mg phenformin drug powder or 12.916 metformin drug powder and 1ml Liquid A is mixed and shaken thoroughly; then 1 ml of liquid B is added and shaken; in this mixed system, the proportion of biguanide drugs is 50 mM, the volume ratio of lysozyme liquid to TCEP liquid is 1:1, and the proportion of biguanide drugs in the final system is 50 mM; the bacterial film is placed in the above mixed system to obtain an external dressing based on phase transition lysozyme loaded with biguanide drugs.

[0027] The primary structure of the protein contains highly fibrillized sequence structures that can cause amyloid accumulation, and its secondary structure contains a large number of α-helical structures frozen by intramolecular disulfide bonds. On this molecular basis, the addition of the highly efficient disulfide bond reducing agent Tris(2-carboxyethyl)phosphine (TCEP) can quickly open the intramolecular disulfide bonds, thereby unlocking the α-helical structure and inducing its spontaneous transformation to a lower-energy β-sheet, thereby forming an amyloid-like assembly. The phase-transition lysozyme-PTL formed by the self-assembly of lysozyme and the highly efficient disulfide bond reducing agent tris(2-carboxyethyl)phosphine can achieve stable interfacial adhesion with various material surfaces through a multi-site adhesion mechanism.

[0028] Biguanides rapidly activate intestinal AMPK and its downstream signaling pathways in the intestine, and then transmit local signals to the central nervous system through the vagus nerve afferent fibers distributed in the intestine, and then control the liver through the vagus nerve efferent fibers, ultimately inhibiting the liver's glucose output. Biguanides inhibit the activity of mGPD, block the α-glycerophosphate shuttle process, cause NADH to accumulate in the cytoplasm, and inhibit the gluconeogenesis process with lactate and glycerol as substrates.

[0029] The structural formula of phenformin contains a benzene ring (lipid soluble). As an aromatic ring, the benzene ring can increase solubility in non-polar solvents and reduce solubility in polar solvents. Phenformin is lipophilic and binds to the inside of the cell membrane with high affinity, so we speculate that phenformin may have a structural advantage in terms of topical effects, which indicates that phenformin may be more easily absorbed and absorbed by skin cells than metformin when used topically.

[0030] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0031] The above description is only a preferred embodiment of the present invention, and does not limit the patent scope of the present invention. All equivalent structural changes made by using the contents of the present invention specification under the inventive concept of the present invention, or directly / indirectly applied in other related technical fields are included in the patent protection scope of the present invention.

[0032] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A phase-shift lysozyme-loaded biguanide drug external dressing and a preparation method thereof, characterized in that: The following steps are involved: S1. Co-cultivate an appropriate amount of kombucha bacteria with tea sugar water; S2, standing at room temperature to cultivate the biofilm; S3. Filter the bacterial membrane with a gauze, rinse with deionized water 3 to 5 times, and centrifuge at 3000 r / min and 4 degrees Celsius for 20 minutes; S4, placing the bacterial film in a 0.1M NaOH solution with a molecular weight of 40 and heating it; S5. Place the biofilm in deionized water and rinse it clean under stirring; S6. Detect the pH value of the biofilm and use filter paper to absorb the moisture of the biofilm; S7, low temperature preservation of biofilm; S8, use 10mM HEPES buffer as solvent; S9, prepare 2 mg / ml Lyz solution A; S10, prepare 50mM TCEP (molecular weight 250) solution B (5M NaOH is used to adjust the pH of TCEP to 6.2); S11, take 24.172 mg of phenformin drug powder or 12.916 mg of metformin drug powder and mix with 1 ml of solution A, and shake thoroughly to mix; S12, add 1 ml of solution B and shake to mix; S13. In this mixed system, the proportion of biguanide drugs is 50 mM; S14, placing the bacterial film in the above mixed system to obtain an external dressing based on phase-shift lysozyme loaded with biguanide drugs.

2. The phase-shift lysozyme-loaded biguanide drug external dressing and its preparation method according to claim 1, characterized in that: The standing time in S2 is 10-15 days, and the thickness of the cultured biofilm is 3 mm.

3. The phase-shift lysozyme-loaded biguanide drug external dressing and its preparation method according to claim 1, characterized in that: The heating temperature in S4 is 90° C. and the heating time is 1 hour.

4. The phase-shift lysozyme-loaded biguanide drug external dressing and its preparation method according to claim 1, characterized in that: The volume ratio of the lysozyme liquid to the TCEP liquid in S13 is 1:1, and the proportion of biguanide drugs in the final system is 50 mM.