Glucoside medicine external preparation for treating diabetic skin wound and preparation method thereof

Abstract

According to the glycoside medicine external preparation for treating diabetic skin wounds and the preparation method of the glycoside medicine external preparation, the prepared external preparation is convenient to use and remarkable in effect, and the clinical administration requirement is met; the external preparation is simple in auxiliary materials and good in stability; the preparation process is simple and can be industrialized on a large scale.

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical preparations, and relates to a glycoside drug external preparation for treating diabetic skin wounds and a preparation method thereof. Background Art

[0002] Diabetic skin trauma is one of the most common serious complications in diabetic patients. The most notable feature of this type of skin trauma is slow healing, accompanied by severe infection, which affects the overall treatment. Severe diabetic skin trauma may even continue to deteriorate into difficult-to-heal ulcers and may bring the risk of amputation. There are currently relatively mature treatment measures for the treatment of simple skin trauma. However, in diabetic patients, metabolic abnormalities lead to skin capillary lesions, causing local skin tissue hypoxia and obstructed nutrient supply. In addition, excessive accumulation of sugar in the traumatized tissue significantly increases the level of reactive oxygen, inflammation, and exogenous bacterial growth, which in turn leads to reduced tissue repair function and reduced local angiogenesis, ultimately significantly delaying the healing of the wound.

[0003] At present, many researchers in this field in China are exploring new treatment strategies based on the special pathological properties of diabetic skin trauma from the aspects of local microenvironment, anti-inflammatory and antibacterial, angiogenesis, and skin re-epithelialization. The early treatment of diabetic skin trauma mainly uses protective dressings, antibiotics, etc. With the development of treatment technology, growth factors that promote angiogenesis and skin healing are gradually applied to the treatment of diabetic skin trauma. Despite this, the actual clinical treatment effect of diabetic skin trauma is still unsatisfactory, and new effective treatment methods need to be further explored.

[0004] Glycoside compound SHPL-49, (2R, 3S, 4S, 5R, 6R)-2-(hydroxymethyl)-6-(4-(4-methoxyphenyl)butoxy)tetrahydro-2H-pyran-3,4,5-triol, the compound structure is as follows:

[0005]

[0006] The previously applied Chinese patent CN109988201B disclosed and authorized the glycoside compound SHPL-49; the Chinese patent CN112625073B disclosed and authorized the preparation method of the glycoside compound; the Chinese patent CN109988202B disclosed and authorized that the glycoside compound can be used to prevent and treat ischemic microcirculatory disorders in the lower limbs. Especially diabetic foot. Studies have found that the glycoside compound can promote the proliferation, migration, and tubule formation of endothelial cells by acting on macrophages, thereby affecting the formation of new blood vessels, while regulating the microenvironment of skin tissue, promoting collagen formation at the skin lesions of diabetic mice, thereby promoting wound healing. Therefore, it provides a reliable guarantee for the research of this molecule in the treatment of ischemic microcirculatory disorders.

[0007] Since glycoside compounds are prone to unstable factors such as oxidation, hydrolysis, and cleavage during the preparation process, how to formulate the glycoside compounds stably and reliably is a difficult problem that needs to be solved urgently. Summary of the invention

[0008] In view of the difficulties in the prior art, the main purpose of the present invention is to provide a glycoside drug external preparation and a preparation method thereof which is easy to use, has significant effects, good stability, simple preparation process and is suitable for large-scale industrial production. Based on this, the present invention is completed.

[0009] In the first aspect, the present invention provides a glycoside drug topical preparation, which is a cream or ointment, and is composed of a glycoside drug active ingredient and a pharmaceutically acceptable excipient. The glycoside drug active ingredient is a glycoside compound SHPL-49, such as (2R, 3S, 4S, 5R, 6R)-2-(hydroxymethyl)-6-(4-(4-methoxyphenyl)butoxy)tetrahydro-2H-pyran-3,4,5-triol as shown in general formula I, and the pharmaceutically acceptable excipients include an aqueous phase, an oil phase, an emulsifier, and an optional wetting agent and / or an antibacterial agent.

[0010]

[0011] Furthermore, the aqueous phase is selected from one or more of propylene glycol (PG), purified water, polyethylene glycol 400 (PEG 400) and / or diethylene glycol monoethyl ether (Transcutol-P).

[0012] Furthermore, the oil phase is selected from one or more of white vaseline, liquid paraffin, beeswax 1014, propylene glycol caprylate (Capryol90), polyethylene glycol 3350 (PEG 3350), mineral oil and / or cetearyl alcohol.

[0013] Furthermore, the emulsifier is selected from one or more of Span 60, Tefose 63, Labrafil M 1944CS and / or Tween 60.

[0014] Furthermore, the optional wetting agent and / or antibacterial agent is selected from one or more of disodium ethylenediaminetetraacetate (EDTA-2Na), glycerol, methylparaben and / or ethylparaben.

[0015] Furthermore, the content of the SHPL-49 compound of formula (I) in the external preparation is 0.5 to 20% by weight, preferably 2 to 10% by weight.

[0016] Furthermore, the content of the water phase in the external preparation is 5-75% by weight, the content of the oil phase is 5-75% by weight, the content of the emulsifier is 5-25% by weight, and the content of the wetting agent and / or antibacterial agent is 0.1-1% by weight.

[0017] In a second aspect, the present invention provides a pharmaceutical composition comprising a glycoside compound, an aqueous phase, an oil phase, an emulsifier and an optional wetting agent and / or an antibacterial agent, wherein the glycoside compound is a compound represented by formula I.

[0018] The aqueous phase is selected from one or more of propylene glycol (PG), purified water, polyethylene glycol 400 (PEG 400) and / or diethylene glycol monoethyl ether (Transcutol-P).

[0019] The oil phase is selected from one or more of white vaseline, liquid paraffin, beeswax 1014, propylene glycol caprylate (Capryol 90), polyethylene glycol 3350 (PEG 3350), mineral oil and / or cetearyl alcohol.

[0020] The emulsifier is selected from one or more of Span 60, Tefose 63, Labrafil M 1944CS and / or Tween 60.

[0021] The optional wetting agent and / or antibacterial agent is selected from one or more of disodium ethylenediaminetetraacetate (EDTA-2Na), glycerol, methylparaben and / or ethylparaben.

[0022] The content of the SHPL-49 compound of formula (I) in the external preparation is 0.5 to 20% by weight, preferably 2 to 10% by weight.

[0023] The external preparation contains 5-75% by weight of the water phase, 5-75% by weight of the oil phase, 5-25% by weight of the emulsifier, and 0.1-1% by weight of the wetting agent and / or antibacterial agent.

[0024] In a third aspect, the present invention provides an application of an external preparation of a glycoside drug in the preparation of an external medicine for treating diabetic skin trauma, wherein the external preparation is a cream or an ointment, which is composed of a glycoside drug active ingredient and a pharmaceutically acceptable excipient, and the glycoside drug active ingredient is a compound shown in formula I.

[0025] In a fourth aspect, a method for preparing a glycoside drug external preparation comprises the following steps:

[0026] S1: Weigh the raw materials: active ingredients of glycoside drugs of formula I, aqueous phase, oil phase, emulsifier and optional wetting agent and / or antibacterial agent:

[0027] S2: Fully stir and dissolve the prescribed amount of glycoside active ingredient SHPL-49 with the aqueous phase;

[0028] S3: Add the prescribed amount of oil phase, emulsifier and optional component wetting agent and / or antibacterial agent to the above aqueous phase solution, heat and stir until melted (70°C, 700rpm), mix evenly and homogenize;

[0029] S4: Continue stirring and cool to room temperature to prepare an external preparation.

[0030] Beneficial Effects

[0031] The advantages of the glycoside drug external preparation of the present invention are: the prepared external preparation is easy to use, has significant effects, and meets the clinical drug administration requirements; the external preparation has simple auxiliary materials and good stability; the preparation process is simple and can be industrialized on a large scale. DETAILED DESCRIPTION

[0032] The specific embodiments of the present invention are further described below. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in the embodiments described below can be combined with each other as long as they do not conflict with each other.

[0033] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.

[0034] Example 1

[0035]

[0036] Preparation process: According to the formula, the active ingredients of glycoside drugs SHPL-49 and PG are fully stirred and dissolved; the prescribed amount of white vaseline, liquid paraffin, Span 60 and methylparaben are added to the above solution, heated and stirred until melted (70°C, 700rpm), mixed evenly and homogenized; continued stirring and cooled to room temperature to prepare a milky white semisolid topical preparation.

[0037] Example 2

[0038]

[0039] Preparation process: According to the formula, the active ingredient of the glycoside drug SHPL-49 is fully stirred and dissolved with purified water; the prescribed amount of Capryol 90, white vaseline, beeswax 1014, Span 60, EDTA-2Na and methylparaben are added to the above solution, heated and stirred until melted (70°C, 700rpm), mixed evenly and homogenized; continued stirring and cooled to room temperature to prepare a milky white semisolid topical preparation.

[0040] Example 3

[0041]

[0042] Preparation process: According to the formula, the active ingredient of the glycoside drug SHPL-49 is fully stirred and dissolved with PG and purified water; the prescribed amount of liquid paraffin, Tefose 63, Labrafil M 1944CS, EDTA-2Na and methylparaben are added to the above solution, heated and stirred until melted (70°C, 700rpm), mixed evenly and homogenized; continued stirring and cooled to room temperature to prepare a milky white semisolid external preparation.

[0043] Example 4

[0044]

[0045] Preparation process: According to the formula, the active ingredient of the glycoside drug SHPL-49, PG and PEG 400 are fully stirred and dissolved; the prescribed amount of PEG 3350, glycerol and methylparaben are added to the above solution, heated and stirred until melted (70°C, 700rpm), mixed evenly and homogenized; continued stirring and cooled to room temperature to prepare a milky white semisolid external preparation.

[0046] Example 5

[0047]

[0048] Preparation process: According to the formula, the active ingredient of the glycoside drug SHPL-49 is fully stirred and dissolved with PG, Transcutol-P and purified water; the prescribed amount of mineral oil, cetearyl alcohol, Tween 60, Span 60, EDTA-2Na and ethylparaben are added to the above solution, heated and stirred until melted (70°C, 700rpm), mixed evenly and homogenized; stirring is continued and cooled to room temperature to prepare a milky white semisolid external preparation.

[0049] Example 6

[0050]

[0051]

[0052] Preparation process: According to the formula, the active ingredient of the glycoside drug SHPL-49 is fully stirred and dissolved with PG and purified water; the prescribed amount of mineral oil, hexadecanol, Tefose 63 (polyethylene glycol-7-stearate), EDTA-2Na and methylparaben are added to the above solution, heated and stirred until melted (70°C, 700 rpm), mixed evenly and homogenized; stirring is continued and cooled to room temperature to prepare a milky white semisolid external preparation.

[0053] Example 7

[0054]

[0055] Preparation process: According to the formula, the active ingredient of the glycoside drug SHPL-49 is fully stirred and dissolved with PG and purified water; the prescribed amount of mineral oil, mono- and distearic glyceryl, hexadecanol, Tefose 63, EDTA-2Na and methylparaben are added to the above solution, heated and stirred until melted (70°C, 700 rpm), mixed evenly and homogenized; stirring is continued and cooled to room temperature to prepare a milky white semisolid external preparation.

[0056] Example 8 Accelerated stability test

[0057] The external preparation samples prepared in Examples 1 to 7 were placed under 40°C and RH75% conditions to conduct accelerated stability tests. The results are as follows:

[0058]

[0059]

[0060]

[0061] The results show that the glycoside drug external preparations of the present invention, the prescriptions of Examples 1-4, have no significant changes in appearance, content and total impurities, and are of stable quality. The prescription of Example 5 has no significant changes in content and total impurities under the conditions of accelerated stability investigation, but the appearance changes slightly at 3 months. Examples 6 and 7 have stable appearance under the conditions of 6-month accelerated stability, but the total impurities increase significantly, and the content of active ingredients decreases significantly, which is presumed to be caused by the degradation of the active ingredients. The above data show that the prescriptions of Examples 1-5 have good chemical stability and physical stability.

[0062] Example 9: Drug efficacy experiment

[0063] 1. Experimental Materials

[0064] Drug preparation: According to the preparation process of Examples 1-5, the drug-containing prescriptions of Prescriptions 1-5 and the corresponding blank control samples were prepared. Experimental animals: C57 BL / 6N mice, SPF grade, male, about 23-25g, 6-7 weeks old, purchased from Shanghai Weitong Lihua Experimental Animal Technology Co., Ltd.

[0065] Feeding conditions: Males and females were kept in separate cages, with no more than 5 mice in each cage. The environmental parameters of the animal room were recorded during the feeding period. Feeding environment: temperature: 20-26℃; relative humidity: 40%-70%; ventilation: ≥15 air changes per hour; feeding feed: SPF rat maintenance feed sterilized by cobalt 60 irradiation, purchased from Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd. Drinking water: free drinking of homemade deionized water. Bedding: corn cob bedding, purchased from Dezhou Gumei Agricultural Technology Co., Ltd.

[0066] 2. Animal model making

[0067] 2.1 Establishment of diabetic mouse model

[0068] This experiment uses C57 BL / 6N male mice, the weight of male mice is controlled at 23g-25g, and the weight is about 25g after normal feed feeding. They are fed with high-sugar and high-fat feed for 3 consecutive weeks. The weight is weighed on the day of modeling, and blood is collected from the tail vein and the basal blood sugar is measured with a blood glucose meter. Before modeling, fasting but not water is allowed for 10 hours, and 1% streptozotocin (STZ) prepared with 0.1mol / L, pH 4.4 citric acid buffer is intraperitoneally injected at 50mg / kg (body weight). STZ is induced for five consecutive days, and the blank control group is injected with an equal volume of citric acid buffer. After that, continue to feed with high-sugar and high-fat feed, and test blood sugar after five days. Mice with blood sugar greater than or equal to 16.7mmol / L are selected as diabetic mice.

[0069] 2.2 Establishment of diabetic mouse skin trauma model

[0070] After the diabetic mice were grouped, a full-thickness skin injury model was prepared on the back skin. The mice were fasted but not watered 12 hours before wound modeling. On the day of modeling, the mice were weighed and blood sugar was measured. The mice were anesthetized with isoflurane breathing anesthesia using a small animal ventilator. The backs of the anesthetized diabetic mice were shaved (slightly above the middle of the lumbar spine), and the surrounding skin was disinfected with 75% ethanol. The 8mm sterile skin biopsy puncher marked the modeling area slightly above the middle of the lumbar spine on the back of the mice. The marked skin and subcutaneous tissue were surgically removed under sterile conditions, avoiding large blood vessels, and paying attention to removing the subcutaneous fascia. The wound was wiped clean with a sterile cotton ball, which was the open wound model of full-thickness skin defect in diabetic mice. After the wound was formed, the mice were kept warm, kept in separate cages, and allowed to drink water and eat high-sugar and high-fat feed freely.

[0071] 3. Dosage regimen

[0072] In the diabetic mouse skin wound model, the wound was smeared with the drug-containing prescriptions of Examples 1-5 and their blank control samples, and the efficacy of continuous administration for 9 days was monitored. The wound surface was photographed with a digital camera on days 0, 3, 6, and 9, and the wound area was counted using PS software to determine the therapeutic effects of SHPL-49 topical preparations with different prescriptions on diabetic skin wounds.

[0073] The experiment required 110 male mice, and a total of 11 groups were set up: diabetic mouse skin wound model group (DM group), DM+drug-containing groups of each prescription of Examples 1-5, and DM+blank control group of Examples 1-5 (drug group that does not contain SHPL-49 active ingredient and is only composed of excipients: Blank group). The dosage of each mouse drug-containing prescription and blank control sample was 0.5 ml / time; the dosage frequency was 2 times / day, and the dosage was continued for 9 days.

[0074] 4. Pharmacodynamic indicators

[0075] Wound area

[0076] In the diabetic mouse wound model, the wounds were smeared with the drug-containing samples of the prescriptions of Examples 1-5 and their blank control samples for 9 consecutive days. Pictures were taken on the 0th, 3rd, 6th and 9th days after the wound was formed, and the wound area was calculated.

[0077] 4.3 Statistical analysis

[0078] GraphPad Prism 6.0 software was used to perform statistical analysis of the data after the experiment. The statistical differences of the data were analyzed by Two-way ANOVA and T-test. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001 indicated significant differences.

[0079] 5. Experimental Results

[0080] Summary of wound area of ​​each group after 9 days of continuous drug administration

[0081]

[0082]

[0083] Note: Two-way ANOVA statistical analysis. *P<0.05, **P<0.01, ***P<0.001DM group vs DM+drug group, DM+drug group vs DM+Blank group. Others are all P>0.05. N: The number of animals treated for 9 consecutive days.

[0084] The experimental results showed that compared with the DM group at the same time point, the drug-containing group of Example 1-5 significantly promoted the healing of the wound surface of diabetic mice on Day 3, 6, and 9 ( * P<0.05, ** P<0.01, *** P<0.001).

[0085] Based on the above data analysis, the external preparations of Examples 1-5 have good chemical and physical stability and can significantly promote the healing of wounds in diabetic mice.

Claims

1. A glycoside drug external preparation, the external preparation is a cream or ointment, composed of a glycoside drug active ingredient and a pharmaceutically acceptable excipient, the glycoside drug active ingredient is a glycoside compound SHPL-49, such as (2R, 3S, 4S, 5R, 6R)-2-(hydroxymethyl)-6-(4-(4-methoxyphenyl)butoxy)tetrahydro-2H-pyran-3,4,5-triol as shown in general formula I, and the pharmaceutically acceptable excipients include an aqueous phase, an oil phase, an emulsifier and an optional wetting agent and / or an antibacterial agent.

2. The preparation according to claim 1, wherein the aqueous phase is selected from one or more of propylene glycol (PG), purified water, polyethylene glycol 400 (PEG400) and / or diethylene glycol monoethyl ether (Transcutol-P); the oil phase is selected from one or more of white vaseline, liquid paraffin, beeswax 1014, propylene glycol caprylate (Capryol 90), polyethylene glycol 3350 (PEG 3350), mineral oil and / or cetearyl alcohol; the emulsifier is selected from one or more of Span 60, Tefose 63, Labrafil M 1944CS and / or Tween 60; the optional wetting agent and / or antibacterial agent is selected from one or more of disodium ethylenediaminetetraacetic acid (EDTA-2Na), glycerol, methylparaben and / or ethylparaben.

3. The preparation according to claim 1, wherein the content of the compound of formula (I) SHPL-49 in the external preparation is 0.5 to 20% by weight, preferably 2 to 10% by weight.

4. The preparation according to claim 1, wherein the content of the water phase in the external preparation is 5 to 75% by weight, the content of the oil phase is 5 to 75% by weight, the content of the emulsifier is 5 to 25% by weight, and the content of the wetting agent and / or antibacterial agent is 0.1 to 1% by weight.

5. A pharmaceutical composition comprising a glycoside compound, an aqueous phase, an oil phase, an emulsifier and an optional wetting agent and / or an antibacterial agent, wherein the glycoside compound is a compound represented by formula I.

6. The composition of claim 5, wherein the aqueous phase is selected from one or more of propylene glycol (PG), purified water, polyethylene glycol 400 (PEG400) and / or diethylene glycol monoethyl ether (Transcutol-P); the oil phase is selected from one or more of white vaseline, liquid paraffin, beeswax 1014, propylene glycol caprylate (Capryol 90), polyethylene glycol 3350 (PEG 3350), mineral oil and / or cetearyl alcohol; the emulsifier is selected from one or more of Span 60, Tefose 63, Labrafil M 1944CS and / or Tween 60; the optional wetting agent and / or antibacterial agent is selected from one or more of disodium ethylenediaminetetraacetic acid (EDTA-2Na), glycerol, methylparaben and / or ethylparaben.

7. The composition according to claim 5, wherein the content of the compound of formula (I) SHPL-49 in the external preparation is 0.5 to 20% by weight, preferably 2 to 10% by weight.

8. The composition according to claim 5, wherein the content of the water phase in the external preparation is 5 to 75% by weight, the content of the oil phase is 5 to 75% by weight, the content of the emulsifier is 5 to 25% by weight, and the content of the wetting agent and / or antibacterial agent is 0.1 to 1% by weight.

9. Use of an external preparation of a glycoside drug in the preparation of an external medicine for treating diabetic skin trauma, wherein the external preparation is a cream or an ointment, which is composed of a glycoside drug active ingredient and a pharmaceutically acceptable excipient, and the glycoside drug active ingredient is a compound shown in formula I.

10. A method for preparing a glycoside drug external preparation, comprising the following steps: S1: Weigh the raw materials: active ingredients of glycoside drugs of formula I, aqueous phase, oil phase, emulsifier and optional wetting agent and / or antibacterial agent: S2: Fully stir and dissolve the prescribed amount of glycoside active ingredient SHPL-49 with the aqueous phase; S3: Add the prescribed amount of oil phase, emulsifier and optional component wetting agent and / or antibacterial agent to the above aqueous phase solution, heat and stir until melted (70°C, 700rpm), mix evenly and homogenize; S4: Continue stirring and cool to room temperature to prepare an external preparation.

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