Gel preparation and preparation method thereof

Abstract

According to the gel preparation and the preparation method thereof provided by the invention, the preparation does not use conventional stabilizers and antioxidants, and the PD-DP-008 gel preparation with better stability is obtained by optimizing the components of the gel preparation. The conventional lofting data of the preparation shows that the content change of the active substance PD-DP-008 is small, and the cumulative release amount can reach 76.005% at most after 8-hour in-vitro release. The prepared gel is uniform and fine in texture, is easy to coat after being unfolded, can form a transparent film on the surface of skin, has the advantages of high adhesiveness, quick drug release, long residence time, no irritation to skin and mucous membrane and the like, and is simple in preparation process, good in antibacterial effect and high in use value.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparations, and in particular to a gel preparation and a preparation method thereof. Background Art

[0002] Acne, also known as acne vulgaris or commonly known as "acne", is the most common chronic inflammatory disease of the hair follicle sebaceous glands in the skin, mainly manifested as acne, papules, nodules and cysts. Its occurrence is closely related to factors such as excessive sebum secretion, blockage of the hair follicle sebaceous gland duct, bacterial infection and inflammatory response. Acne is prone to occur on the face and upper trunk of adolescents. After entering this period, the level of androgen in the body rises rapidly, promoting the development of sebaceous glands and producing a large amount of sebum. At the same time, the abnormal keratinization of the hair follicle sebaceous gland duct causes duct blockage, sebum discharge is impaired and acne is formed. The current difficulties in the treatment of acne in clinical practice are drug resistance and easy recurrence.

[0003] At present, the treatment of acne at home and abroad is still mainly based on classic therapeutic drugs, such as retinoic acid, benzoyl peroxide and antibiotics, and the dosage forms are mostly topical semi-solid preparations or oral solid preparations. Compared with systemic medications, local medication preparations have obvious advantages in treating acne. They can not only avoid or reduce systemic adverse reactions, but also avoid the first-pass effect of drugs in the liver, and the dosage used for local effectiveness of the preparations is significantly reduced.

[0004] This application is based on the extended research of the antimicrobial peptide PD-DP-008 (CN 110627870 A, CN114209808A) discovered by the company in the early stage of research and development. This product is designed for acne caused by Propionibacterium acnes and has a good therapeutic effect on drug-resistant Propionibacterium acnes. The dosage form is developed as an external gel in order to solve the problem of drug-resistant acne that cannot be cured for a long time. Summary of the invention

[0005] The purpose of the present invention is to provide a gel preparation and a preparation method thereof in view of the above-mentioned deficiencies of the prior art, so as to increase the release rate of the gel, increase transdermal absorption, improve the stability of the preparation, and make the formulation process design stable and suitable for commercial production.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] The first object of the present invention is to provide a gel preparation comprising:

[0008] (a) PD-DP-008, the amino acid sequence of which is shown in SEQ ID NO.1,

[0009] (b) Hydroxypropyl methylcellulose,

[0010] (c) Propylene glycol.

[0011] The second object of the present invention is a gel preparation comprising:

[0012] (a) PD-DP-008, the amino acid sequence of which is shown in SEQ ID NO.1,

[0013] (b) Hydroxypropyl methylcellulose,

[0014] (c) propylene glycol,

[0015] (d) Dimethyl sulfoxide.

[0016] Furthermore, the mass ratio of PD-DP-008 to hydroxypropyl methylcellulose is 0.1:(1-3).

[0017] Furthermore, the mass ratio of hydroxypropyl methylcellulose to propylene glycol is (1-3):(5-20).

[0018] Furthermore, the mass ratio of PD-DP-008, hydroxypropyl methylcellulose and propylene glycol is 0.1:(2-3):(10-20).

[0019] Furthermore, the mass ratio of hydroxypropyl methylcellulose to dimethyl sulfoxide is (1-3):8.

[0020] Further, based on 100 parts by total weight, the composition is as follows:

[0021] PD-DP-008 0.1 part;

[0022] 2-3 parts of Hydroxypropyl Methylcellulose;

[0023] Propylene glycol 5-20 parts;

[0024] 8 parts of dimethyl sulfoxide;

[0025] The balance is water.

[0026] Further, based on 100 parts by total weight, the composition is as follows:

[0027] PD-DP-008 0.1 part;

[0028] 2 parts of Hydroxypropyl Methylcellulose;

[0029] Propylene glycol 10 parts;

[0030] 8 parts of dimethyl sulfoxide;

[0031] The balance is water.

[0032] The third object of the present invention is to provide a method for preparing the above-mentioned gel preparation, comprising the following steps:

[0033] S1. After emulsifying the hypromellose at high speed, add a prescribed amount of propylene glycol, mix well, and obtain a matrix;

[0034] S2. Weigh the prescribed amount of PD-DP-008, dissolve it evenly with purified water, add it to the matrix while stirring, and continue stirring to obtain PD-DP-008 gel.

[0035] Furthermore, step S1 also includes adding a prescribed amount of dimethyl sulfoxide.

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] (1) The present invention provides a pharmaceutical preparation for treating acne which is easy to use, has a controlled release effect, and has little toxicity or side effects.

[0038] (2) The gel preparation provided by the present invention does not use conventional stabilizers and antioxidants, but optimizes the components of the gel preparation to obtain a PD-DP-008 gel preparation with controlled release and good transdermal absorption. Moreover, the accelerated sampling data of the preparation shows that the content of the active substance PD-DP-008 changes little, and the cumulative release amount in vitro for 8 hours can reach up to 76.005%.

[0039] (3) The auxiliary materials of the present invention use propylene glycol as a solubilizer and moisturizer, and dimethyl sulfoxide DMSO as a transdermal absorption enhancer, which reduces the irritation to the skin caused by the use of alcohol in the traditional process, and the specific proportion of propylene glycol, DMSO and hydroxypropyl methylcellulose HPMC can effectively improve the effect of PD-DP-008 gel in treating acne. The prepared gel has a uniform and fine texture, is easy to apply after unfolding, can form a transparent film on the skin surface, has the advantages of strong adhesion, fast drug release, long retention time, and no irritation to the skin and mucous membranes. The process of the present invention is simple, the product has a good antibacterial effect, and has a good use value.

[0040] (4) The present invention constructs a mouse model and uses fusidic acid cream as a positive control to verify that the PD-DP-008 gel preparation has a significant therapeutic effect on acne vulgaris. DETAILED DESCRIPTION

[0041] In order to make the purpose, technical scheme and advantages of the present invention clearer, the specific embodiments of the present invention are further described in detail below in conjunction with specific examples. If no specific technology or conditions are specified in the examples, the technology or conditions described in the literature in this field or the product instructions are used. If the manufacturer of the reagents or instruments is not specified, they are all conventional products that can be obtained commercially.

[0042] Example 1

[0043] Preparation of different formulations of PD-DP-008 gel (specification: 0.1% (10 g: 10 mg)).

[0044] (1) The compositions of Prescription 1 and Prescription 2 are shown in Table 1.

[0045] Table 1. Composition of Formulation 1 and Formulation 2.

[0046]

[0047] Preparation process:

[0048] In a 100ml beaker, add an appropriate amount of purified water (prescription 1: 39.1g, prescription 2: 35.1g), 5.0g of propylene glycol, and an appropriate amount of DMSO (prescription 1: 0g, prescription 2: 4g), stir to dissolve evenly, add 0.5g of carbomer while stirring, continue stirring until the carbomer powder is completely swollen, add 0.4g of triethanolamine while stirring at a low speed to make a uniform matrix 1 and 2, set aside.

[0049] Weigh two portions of PD-DP-008, 62.5 mg each, dissolve each in 5 g of water, add each to matrix 1 and 2 while stirring, stir evenly, and the mixture is obtained.

[0050] (2) The compositions of Prescription 3 and Prescription 4 are shown in Table 2.

[0051] Table 2. Composition of Formulation 3 and Formulation 4.

[0052]

[0053] Preparation process:

[0054] In a 100ml beaker, add about 62.5mg of PD-DP-008, an appropriate amount of purified water (prescription 3: 44.1g, prescription 4: 40.1g), 5.0g of propylene glycol, and an appropriate amount of DMSO (prescription 3: 0g, prescription 4: 4g), stir to dissolve evenly, add 0.5g of carbomer while stirring, continue stirring until the carbomer powder is completely swollen, add 0.4g of triethanolamine while stirring at a low speed, and continue stirring to form a uniform gel.

[0055] (3) The compositions of Prescription 5 and Prescription 6 are shown in Table 3.

[0056] Table 3. Composition of Formulation 5 and Formulation 6.

[0057]

[0058] Preparation process:

[0059] Heat purified water to 80-90°C, weigh an appropriate amount (prescription 5: 38.5g, prescription 6: 34.5g) into a 100ml beaker, add 1.5g of hydroxypropyl methylcellulose, 5.0g of propylene glycol, and an appropriate amount of DMSO (prescription 5: 0g, prescription 6: 4g) while stirring, continue stirring until cooled, and obtain matrix 5 and 6 for use.

[0060] Weigh two portions of PD-DP-008, 62.5 mg each, dissolve each in 5 g of water, add each to matrix 5 and 6 while stirring, stir evenly, and obtain the product.

[0061] (4) The compositions of Prescription 7 and Prescription 8 are shown in Table 4.

[0062] Table 4. Composition of Formulation 7 and Formulation 8.

[0063]

[0064] Preparation process:

[0065] In a 250ml beaker, add 100ml of purified water and 5.0g of sodium carboxymethyl cellulose, place in the refrigerator overnight to swell, stir evenly, take 35.05g of swollen sodium carboxymethyl cellulose, 6.45g of propylene glycol, and an appropriate amount of DMSO (prescription 7: 0g, prescription 8: 4g), continue stirring to make uniform matrices 7 and 8, and set aside.

[0066] Weigh two portions of PD-DP-008, 62.5 mg each, dissolve prescription 7 in 8.5 g of water and prescription 8 in 4.5 g of water, add them to matrix 7 and 8 respectively while stirring, stir evenly, and the mixture is obtained.

[0067] Example 2

[0068] The in vitro release rates of different formulations were investigated.

[0069] (1) Solution preparation and determination method

[0070] Test solution: Take about 0.2g of the product, weigh accurately, and evenly apply it on a 0.45μm PES filter membrane. Use the Franz diffusion cell method, with water as the receiving medium, the diffusion cell temperature at 32±1℃, the rotation speed at 600r / min, and the sampling time points at 2h, 4h, 6h, and 8h. Take 1.0ml of sample each time.

[0071] Reference substance solution: Take an appropriate amount of PD-DP-008 reference substance, weigh accurately, dissolve in water and quantitatively dilute to make a solution containing about 0.025 mg per 1 ml, and shake well.

[0072] Determination method: Accurately measure the test solution and control solution, inject them into liquid chromatograph respectively, and record the chromatogram.

[0073] (2) Test method: Chromatographic column: octadecylsilane bonded silica gel as filler (C18 5μm150×4.6mm)

[0074] Mobile phase A: 70mmol / L sodium perchlorate (pH adjusted to 2.8 with phosphoric acid)

[0075] Mobile phase B: Mobile phase A-acetonitrile (20:80)

[0076] Detection wavelength: 220nm; flow rate: 1.0ml / min; column temperature: 30℃; injection volume: 100μl

[0077] Diluent: Water

[0078] Perform gradient elution according to Table 5:

[0079] Table 5 Elution schedule.

[0080] Time (min) A％ B％ 0 60 40 15 40 60 16 60 40 20 60 40

[0081] (3) Calculation formula

[0082]

[0083] Cumulative release (%) = Xi + (X1 + X2 + ... + Xi-1) × 1.0 / 8

[0084] The in vitro release rates of different formulations are shown in Table 6:

[0085] Table 6. Statistics of in vitro release of different formulations.

[0086]

[0087] As shown in Table 6, when four materials, namely, carbomer deionized type (2020 type), carbomer homopolymer type B, sodium carboxymethyl cellulose and hypromellose, are used as gel matrices, only when hypromellose is used as the matrix, the sample is released. The hypromellose may be any one of hypromellose E10M, hypromellose E4M and hypromellose K4M.

[0088] Example 3

[0089] A study to evaluate the efficacy of different formulations of PD-DP-008 gel.

[0090] (1) Experimental methods

[0091] A total of 72 SPF SD rats that passed quarantine were selected, half male and half female, and divided into a normal control group (6 rats) and a model group (66 rats). The model group rats were smeared with 2% coal tar once a day on the inner side of the right ear of the ear tube opening, and 6×10 8CFU / mL Propionibacterium acnes bacterial liquid (strain number: ATCC11827, item number: BNCC336443), 3 weeks after modeling, if the right ear modeling site appears red, swollen and desquamated, and dry and nodular when touched, it indicates that the rat auricle complex acne model has been successfully replicated; 60 model rats with obvious redness and swelling symptoms were selected and randomly divided into model control group, fusidic acid ointment group, PD-DP-008 gel prescription 1, 2, 3, 4, 5, 6, 7, and 8 groups according to the thickness of the auricle, with 6 rats in each group; each group of drugs was evenly applied to the skin lesions of the right ear of the rat, with a range of about 1cm×1cm and a thickness of about 1mm, 40mg / rat, twice a day, for 7 consecutive days; the thickness of the auricle of each group was measured before administration, 3 days after administration, and 7 days after administration, and the auricle swelling rate was calculated. Swelling rate = (auricle swelling after administration-auricle swelling before administration) / auricle swelling before administration.

[0092] (2) Experimental results

[0093] As shown in Table 7, compared with the normal control group, the auricle swelling degree and swelling rate of the rats in the model control group after 3-day administration were significantly increased (P≤0.01); compared with the model control group, the auricle swelling degree and swelling rate of the rats in the PD-DP-008 gel prescription group 1, PD-DP-008 gel prescription group 2, PD-DP-008 gel prescription group 3, PD-DP-008 gel prescription group 5, PD-DP-008 gel prescription group 6, and fusidic acid cream group after 3-day administration were significantly reduced (P≤0.05 or P≤0.01); the auricle swelling degree of the PD-DP-008 gel prescription group 7 after 3-day administration was significantly reduced (P≤0.05).

[0094] Compared with the normal control group, the auricle swelling degree and swelling rate of the rats in the model control group after 7 days of administration were significantly increased (P≤0.01); compared with the model control group, the auricle swelling degree and swelling rate of the rats in the PD-DP-008 gel prescription 6 group and the fusidic acid cream group after 7 days of administration were significantly reduced (P≤0.05 or P≤0.01).

[0095] Table 7 Effect of PD-DP-008 on the swelling degree and swelling rate of rat auricle ( n=6)

[0096]

[0097]

[0098] Note: Compared with the normal control group, ++ P≤0.01; compared with the model control group, * P≤0.05, ** P≤0.01.

[0099] Combined with the results of in vitro release test and efficacy test, it is proved that HPMC is more suitable for the matrix of PD-DP-008 than carbomer and sodium carboxymethyl cellulose.

[0100] Example 4

[0101] Investigate the amount of propylene glycol used in the prescription.

[0102] (1) Prescription process design

[0103] Prescription 9: Specification: 0.1% (10g: 10mg)

[0104] Table 8. Propylene glycol dosage screening - prescription 9

[0105]

[0106] Heat purified water to 80-90°C, weigh 160 g into a 250 ml beaker, add 4 g of hypromellose, 10 g of propylene glycol, and 16 g of DMSO while stirring, continue stirring until cooled (the hypromellose is well swollen), and set aside.

[0107] Weigh 95 g of the matrix for later use, take about 124.2 mg of PD-DP-008, dissolve it in 5 g of water, add it to the matrix while stirring, stir evenly, and you have it.

[0108] Add 5 g of purified water to the remaining matrix and stir evenly as a blank excipient.

[0109] Prescription 10: Specification: 0.1% (10g: 10mg)

[0110] Table 9. Propylene glycol dosage screening - prescription 10

[0111]

[0112] Heat purified water to 80-90°C, weigh 150g into a 250ml beaker, add 4g of hypromellose, 20g of propylene glycol, and 16g of DMSO while stirring, continue stirring until cooled (the hypromellose is well swollen), and set aside.

[0113] Weigh 95 g of the matrix for later use, take about 124.2 mg of PD-DP-008, dissolve it in 5 g of water, add it to the matrix while stirring, stir evenly, and you have it.

[0114] Add 5 g of purified water to the remaining matrix and stir evenly as a blank excipient.

[0115] Prescription 11: Specification: 0.1% (10g: 10mg)

[0116] Table 10. Propylene glycol dosage screening - prescription 11

[0117]

[0118] Heat purified water to 80-90°C, weigh 130 g into a 250 ml beaker, add 4 g of hypromellose, 40 g of propylene glycol, and 16 g of DMSO while stirring, continue stirring until cooled (the hypromellose is well swollen), and set aside.

[0119] Weigh 95 g of the matrix for later use, take about 124.2 mg of PD-DP-008, dissolve it in 5 g of water, add it to the matrix while stirring, stir evenly, and you have it.

[0120] Add 5 g of purified water to the remaining matrix and stir evenly as a blank excipient.

[0121] (2) Sample testing

[0122] The prepared samples were tested for properties, related substances, viscosity, and release curve, and the remaining samples were placed under light (light intensity 5000 Lux, ultraviolet near energy 90 μw / cm 2 The total illumination for 10 days is about 1.2×10 6 Lux﹒ hr, ultraviolet energy is about 216w﹒ hr / m 2 ), high temperature 40℃ and 60℃ tests were carried out to examine the changes in related substances and physical and chemical properties.

[0123] ①Related substances analysis methods

[0124] Chromatographic conditions: chromatographic column phenomenex Luna@Omega 5μm PS C18 4.6×250mm

[0125] Mobile phase A: 70mmol / L sodium perchlorate (pH 2.8)

[0126] Mobile phase B: Mobile phase A-acetonitrile (20:80)

[0127] Column temperature: 30°C; wavelength: 220nm; flow rate: 0.9ml / min; injection volume: 100μl

[0128] Elution was performed with the following gradient program:

[0129] Time (min) A％ B％ 0 62 38 50 40 60 55 25 75 60 25 75 61 62 38 70 62 38

[0130] Solution preparation: weigh about 1g of gel sample into a beaker, add an appropriate amount of 50% methanol solution, mix well, transfer the entire amount to a 20ml volumetric flask, make up to volume with 50% methanol, shake well, filter using a nylon filter head, discard 1ml, and take the filtrate as the test solution.

[0131] ② In vitro release analysis method: same as Example 2.

[0132] Solution preparation and determination method:

[0133] Test solution: Take about 0.2g of the product, weigh accurately, and evenly apply it on a 0.45μm PES filter membrane. Use the Franz diffusion cell method, with water as the receiving medium, the diffusion cell temperature at 32±1℃, the rotation speed at 600 rpm, and the sampling time points at 0.5h, 1h, 2h, 3h, 4h, and 6h. Take 1.0ml of sample each time.

[0134] Reference substance solution: Take an appropriate amount of PD-DP-008 reference substance, weigh accurately, dissolve in water and quantitatively dilute to make a solution containing about 0.025 mg per 1 ml, and shake well.

[0135] Determination method: Accurately measure the test solution and control solution, inject them into liquid chromatograph respectively, and record the chromatogram.

[0136] (3) Test results

[0137] Table 11. In vitro release rate of propylene glycol dosage formulation screening

[0138]

[0139] Table 12. Stability study of propylene glycol dosage prescription screening

[0140]

[0141] (4) Conclusion

[0142] The research results with propylene glycol dosage of 5%, 10% and 20% respectively showed that with the increase of propylene glycol, the viscosity of the product increased accordingly; the in vitro cumulative release rate of the sample also increased slightly, but the effect was not obvious; there was no significant effect on the stability of the sample, so the dosage of propylene glycol was selected as the conventional dosage of 10%.

[0143] Example 5

[0144] The stability of PD-DP-008 gel formulations was evaluated.

[0145] PD-DP-008 gel preparation stability test method: The gel was placed in a stability test box at 25±2°C, RH65±5%. After the initial inspection (0 day), the stability of the sample was inspected and relevant data were collected at sampling inspection time points of 1 month, 2 months, 3 months and 6 months.

[0146] During the accelerated test period from 0 to 6 months, the maximum single impurity of the PD-DP-008 gel sample changed from 0.48% to 1.94%, and the total impurity increased from 1.98% to 5.40%, with a 4.4% decrease in content. In addition, the appearance and acidity indicators were basically consistent with those on day 0, with no significant changes, and the gel sample was relatively stable. As shown in Table 13 below.

[0147] Table 13. Summary of data on accelerated stability study of PD-DP-008 gel.

[0148]

[0149] For matters not mentioned above, the prior art applies.

[0150] Although some specific embodiments of the present invention have been described in detail through examples, those skilled in the art should understand that the above examples are for illustration only and are not intended to limit the scope of the present invention. Those skilled in the art to which the present invention belongs may make various modifications or supplements to the specific embodiments described or replace them in a similar manner, but they will not deviate from the direction of the present invention or exceed the scope defined by the attached claims. Those skilled in the art should understand that any modifications, equivalent substitutions, improvements, etc. made to the above embodiments based on the technical essence of the present invention should be included in the protection scope of the present invention.

Claims

1. A gel preparation, characterized in that: contain: (a) PD-DP-008, the amino acid sequence of which is shown in SEQ ID NO.1, (b) Hydroxypropyl methylcellulose, (c) Propylene glycol.

2. A gel preparation, characterized in that: contain: (a) PD-DP-008, the amino acid sequence of which is shown in SEQ ID NO.1, (b) Hydroxypropyl methylcellulose, (c) propylene glycol, (d) Dimethyl sulfoxide.

3. The gel preparation according to any one of claims 1 to 2, characterized in that The mass ratio of PD-DP-008 to hydroxypropyl methylcellulose is 0.1:(1-3).

4. The gel preparation according to any one of claims 1 to 2, characterized in that The mass ratio of hydroxypropyl methylcellulose to propylene glycol is (1-3):(5-20).

5. The gel preparation according to any one of claims 1 to 2, characterized in that The mass ratio of PD-DP-008, hydroxypropyl methylcellulose and propylene glycol is 0.1:(2-3):(10-20).

6. The gel preparation according to claim 2, characterized in that The mass ratio of hydroxypropyl methylcellulose to dimethyl sulfoxide is (1-3):

8.

7. The gel preparation according to claim 2, characterized in that Based on 100 parts by total weight, the composition is as follows: PD-DP-008 0.1 part; 2-3 parts of Hydroxypropyl Methylcellulose; Propylene glycol 5-20 parts; 8 parts of dimethyl sulfoxide; The balance is water.

8. The gel preparation according to claim 2, characterized in that Based on 100 parts by total weight, the composition is as follows: PD-DP-008 0.1 part; 2 parts of Hydroxypropyl Methylcellulose; Propylene glycol 10 parts; 8 parts of dimethyl sulfoxide; The balance is water.

9. A method for preparing the gel preparation according to claim 1, characterized in that: The following steps are involved: S1. After emulsifying the hypromellose at high speed, add a prescribed amount of propylene glycol, mix well, and obtain a matrix; S2. Weigh the prescribed amount of PD-DP-008, dissolve it evenly with purified water, add it to the matrix while stirring, and continue stirring to obtain PD-DP-008 gel.

10. The preparation method according to claim 9, characterized in that: Step S1 also includes adding a prescribed amount of dimethyl sulfoxide.

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