External emulsifiable paste capable of relieving post-traumatic bone joint degeneration pain symptoms and preparation method of external emulsifiable paste

Abstract

The invention relates to the field of biological medicine, and discloses external cream capable of relieving post-traumatic bone joint degeneration pain symptoms and a preparation method of the external cream. By constructing a posttraumatic osteoarthritis rat model, it is found that experimental rats applying the atractylenolide I ointment group are superior to a control group in the aspects of behavioral analysis, bone mineral content, pain conduction and the like. According to the research, the suspension latex preparation containing atractylenolide particles is prepared from the Chinese herbal medicine extract atractylenolide I through an advanced medicine preparation process for the first time and is used for epidermis joints of lower limbs and the like. The composition can be used for improving arthralgia symptoms caused by joint degeneration after trauma, and an experimental foundation is laid for follow-up research of related drugs.

Description

Technical Field

[0001] The present application relates to the field of biomedicine, and more specifically, to an external-use cream capable of relieving pain symptoms of post-traumatic bone and joint degeneration and a preparation method thereof. Background Art

[0002] With an aging population and increasing sports injuries, the incidence of bone and joint degeneration, particularly post-traumatic degeneration, is increasing annually. This post-traumatic degeneration can cause limb pain and even disability. In later stages, most patients require joint replacements, significantly increasing healthcare costs. Various interventions can alleviate pain, slow disease progression, improve patients' quality of life, and maximize limb function.

[0003] By leveraging Traditional Chinese Medicine's Zang-Xiang theory and the Five Elements' properties as described in the Yellow Emperor's Classic of Internal Medicine, a new approach to alleviating pain caused by post-traumatic bone and joint degeneration can be found by regulating the spleen. Atractylenolide I, a traditional spleen-tonifying herb, has been shown to possess anti-inflammatory properties. It is the primary active ingredient in Atractylenolide I, a herb used to strengthen the spleen.

[0004] However, whether Atractylodes lactone I can play a role in relieving joint pain and improving osteoporosis has not been reported. Summary of the Invention

[0005] The purpose of the invention of this application is to explore the role of Atractylodes lactone I in relieving joint pain and improving osteoporosis, and to provide an external cream that can relieve the pain symptoms of post-traumatic bone and joint degeneration and its preparation method.

[0006] In order to achieve the above-mentioned invention objectives, this application adopts the following technical solutions:

[0007] In a first aspect, the present application discloses the use of Atractylodes lactone I in the preparation of drugs for relieving joint pain and improving osteoporosis.

[0008] Furthermore, the joint pain refers to joint pain caused by post-traumatic joint degeneration.

[0009] In a second aspect, the present application discloses a drug having Atractylodes lactone I as an active ingredient.

[0010] Furthermore, the medicine also includes pharmaceutically acceptable excipients.

[0011] Furthermore, the dosage form of the drug is an external cream.

[0012] In a third aspect, the present application discloses an external-use cream capable of relieving pain symptoms of post-traumatic bone and joint degeneration, comprising, by weight:

[0013] Atractylodes lactone Ⅰ 0.1-1 part;

[0014] 0.1-1 part of octadecyl alcohol;

[0015] 0.1-1 part of white vaseline;

[0016] 0.1-1 parts of glyceryl monostearate;

[0017] 0.1-1 part of glycerin;

[0018] Emulsifier 0.1-1 part;

[0019] 1 to 5 parts of distilled water.

[0020] In a fourth aspect, the present application discloses a method for preparing the topical cream, comprising the following steps:

[0021] Dissolve the emulsifier and glycerol in distilled water, add Atractylodes lactone I and mix thoroughly by ultrasonic dispersion to form a suspension, heat in a water bath to 50-90°C and incubate for 3-9 minutes to obtain an aqueous phase;

[0022] Separately, take stearyl alcohol, white vaseline, and glyceryl monostearate and heat them on a water bath until they are completely dissolved and the temperature is between 50 and 90°C to obtain an oil phase;

[0023] The oil phase is slowly added to the water phase while continuing to stir in the same direction until condensation occurs, thereby obtaining the external cream.

[0024] In summary, this application has the following beneficial effects:

[0025] This application establishes a rat model of post-traumatic osteoarthritis (left knee anterior cruciate ligament transection (ACLT)) and finds that the rats treated with Atractylodes lactone I ointment outperformed the control group in behavioral analysis, bone mineral content, and pain transmission. This study is the first to utilize the Chinese herbal extract Atractylodes lactone I using advanced pharmaceutical preparation technology to create a suspension latex formulation containing Atractylodes lactone particles for use in the epidermal joints of the lower limbs. This formulation can be used to improve joint pain symptoms caused by post-traumatic joint degeneration, laying the experimental foundation for subsequent research on related drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 : Behavioral analysis results in Example 2 of the present application;

[0027] Figure 2 : The bone density test results in Example 3 of the present application;

[0028] Figure 3 : Immunofluorescence results in Example 4 of the present application;

[0029] Figure 4: Virus retrograde tracing results in Example 4 of the present application. DETAILED DESCRIPTION

[0030] The technical solutions and effects of the present application are further described in detail below with reference to the embodiments and drawings. It should be understood that the specific embodiments described herein are only used to explain the invention, rather than to limit the invention.

[0031] Example 1: Preparation of Atractylodes lactone I cream

[0032] This embodiment discloses an external-use cream capable of relieving pain symptoms of post-traumatic bone and joint degeneration. The raw materials used for its preparation include: 1 g of Atractylodes lactone I, 1 g of octadecyl alcohol, 1 g of white vaseline, 1 g of glyceryl monostearate, 1 g of glycerol, 1 g of emulsifier, and 5 g of distilled water.

[0033] The preparation method comprises the following steps:

[0034] Take the emulsifier and glycerol and dissolve them in distilled water, add atractylodes lactone I and mix them by ultrasonic dispersion to form a suspension, heat it in a water bath to 60℃ and incubate for 5 minutes to obtain an aqueous phase;

[0035] Separately, stearyl alcohol, white vaseline, and glyceryl monostearate were heated on a water bath until completely dissolved and the temperature reached 60°C to obtain an oil phase;

[0036] The oil phase is slowly added to the water phase while continuing to stir in the same direction until condensation occurs, thereby obtaining the external cream.

[0037] Example 2: Atractylodes lactone I alleviates pain behavior in osteoarthritis model rats

[0038] Model building

[0039] A rat model of post-traumatic osteoarthritis (left anterior cruciate ligament transection (ACLT)) was established. Eighteen 4-month-old male SD rats were divided into three groups (n=6): a sham-operated (normal control) group, a positive control group, and an atractylodes lactone I-treated group. The rats in the atractylodes lactone I-treated group were treated starting on day 4 after modeling with 3 mg of atractylodes lactone I cream applied to the affected knee joint daily, morning and evening, until day 45 after surgery. The sham-operated and positive control groups received no treatment. Behavioral analysis was performed 3 days before modeling and at 1, 2, 3, 4, 5, and 6 weeks after modeling. Samples were collected and analyzed histologically at week 6.

[0040] Behavioral analysis

[0041] The mechanical pain threshold of rats with osteoarthritis was detected by von Frey filament test and weight-bearing ratio of affected limb.

[0042] For the von Frey filament test, rats are placed in a transparent plastic cylinder (20 cm diameter) with a wire mesh bottom and allowed to acclimate for 15 minutes. One of eight von Frey filaments with stiffnesses ranging from 0.41 to 15.14 g is used. One end is pressed against the plantar skin, pushing the fiber until it bends slightly. A positive response is considered a withdrawal of the hind paw. The interval between stimulations is 10 seconds. A lower pain threshold indicates more severe pathological pain in the rat.

[0043] To test the weight-bearing rate of the affected limb, perform adaptive training once daily for three consecutive days before testing. Rats were placed in a bipedal balance test box and acclimated for 30 minutes in a standardized posture, with both hind legs resting upright on the pressure sensors of the tester and the forelimbs resting on the inclined board in front. The average of the weight bearing on the left and right legs was continuously measured over 5 seconds. This measurement was repeated three times, with a 1-minute interval between readings, and the average of the three readings was used. Weight-bearing rate of the affected limb = (weight bearing on the left hind limb / total weight bearing on both hind limbs) × 100%.

[0044] Behavioral testing was performed on the rat model of post-traumatic osteoarthritis. Figure 1 As shown, we can see that:

[0045] Hindlimb weight-bearing of the ipsilateral limb: Compared with the control group, the hindlimb weight-bearing of the Atractylodes macrocephala-1 group was significantly improved, and the differences were statistically significant. At week 6 after modeling, the hindlimb weight-bearing of the Atractylodes macrocephala-1 group had returned to the level of the sham operation group.

[0046] Paw withdrawal threshold of rats: Compared with the positive control group, the paw withdrawal threshold of the Atractylodes macrocephala lipid-1 group was significantly increased starting from the third week after modeling, and the differences were statistically significant.

[0047] Example 3: Atractylodes lactone I improves osteoporosis in osteoarthritis model rats

[0048] Model building

[0049] A rat model of post-traumatic osteoarthritis (left anterior cruciate ligament transection (ACLT)) was established. Eighteen 4-month-old male SD rats were divided into three groups (n=6): a sham-operated (normal control) group, a positive control group, and an atractylodes lactone I-treated group. The rats in the atractylodes lactone I-treated group were treated starting on day 4 after modeling with 3 mg of atractylodes lactone I cream applied to the affected knee joint daily, morning and evening, until day 45 after surgery. The sham-operated and positive control groups received no treatment. Behavioral analysis was performed 3 days before modeling and at 1, 2, 3, 4, 5, and 6 weeks after modeling. Samples were collected and analyzed histologically at week 6.

[0050] Bone density testing

[0051] After anesthetizing the rats, bone density was measured using dual-energy X-ray absorptiometry. Detection parameters included bone mineral density (BMD) of the whole body and knee joint region of interest, bone mineral content (BMC), bone surface area, tissue surface area, muscle weight, fat weight, and total weight.

[0052] The BMD, BMC, bone surface area, tissue surface area, muscle weight, fat weight, total weight, and ROL area of ​​the left knee joint (modeling) and right knee joint (normal) of the three groups of rats were measured. Figure 2 As shown, we can see that:

[0053] Compared with the positive control group, the bone density-related parameters in the Atractylodes lactone 1 group were improved, and the differences were statistically significant; there was no statistical significance in BMD measurement.

[0054] Example 4: Atractylodes lactone I alleviates pain symptoms in osteoarthritis model rats

[0055] Model building

[0056] A rat model of post-traumatic osteoarthritis (left anterior cruciate ligament transection (ACLT)) was established. Eighteen 4-month-old male SD rats were divided into three groups (n=6): a sham-operated (normal control) group, a positive control group, and an atractylodes lactone I-treated group. The rats in the atractylodes lactone I-treated group were treated starting on day 4 after modeling with 3 mg of atractylodes lactone I cream applied to the affected knee joint daily, morning and evening, until day 45 after surgery. The sham-operated and positive control groups received no treatment. Behavioral analysis was performed 3 days before modeling and at 1, 2, 3, 4, 5, and 6 weeks after modeling. Samples were collected and analyzed histologically at week 6.

[0057] Dorsal root ganglion neuron labeling and detection

[0058] Five weeks after modeling, the virus was injected into the knee joint cavity. Six weeks after modeling, the rats' spinal cords and dorsal root ganglia (DRGs) were removed, frozen, sliced ​​into 20-μm sections, and stored at -20°C until further immunofluorescence analysis. The number of calcitonin gene-related peptide (CGRP)-positive neurons in the spinal cord and DRG tissues was counted under a fluorescence microscope.

[0059] (1) Immunofluorescence results showed that compared with the positive control group, the calcitonin gene-related peptide (CGRP) in the dorsal root ganglion (DRG) of rats in the atractylodes lactone I group was significantly decreased.

[0060] (2) The three groups of animals were injected with a retrograde tracer (viral vector) in the left knee joint. Specifically, after the rats were anesthetized, the skin of the affected hind knee joint was prepared and disinfected with iodine. The knee joint was flexed, and the insulin needle was inserted from the outermost side of the patellar tendon under the patella. The needle was stopped when a clear sense of missing was felt (the insertion depth can be appropriately marked to keep it relatively consistent each time). Then, the tracer was slowly injected into the affected joint cavity with an injection volume of 5μl PRV (BrainVTA). The animals were placed back in the cage for observation. One week later, the viral signal in the spinal cord and DRG was detected to label the sensory neurons.

[0061] The results showed that compared with the positive control group, the sensory nerve innervation in the spinal cord of the Atractylodes macrolide I group was significantly reduced. The viral retrograde tracing results of DRG also showed that compared with the positive control group, the sensory nerve innervation in the Atractylodes macrolide I group was significantly reduced.

[0062] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make non-creative modifications to the present embodiment as needed, but as long as they are within the scope of the claims of the present application, they are protected by the patent law.

Claims

1. Application of Atractylodes lactone I in the preparation of drugs for relieving joint pain and improving osteoporosis.

2. The use according to claim 1, characterized in that The joint pain refers to joint pain caused by post-traumatic joint degeneration.

3. A drug, characterized in that The medicine uses atractylodes lactone I as an active ingredient.

4. The drug according to claim 3, characterized in that Pharmaceutically acceptable excipients are also included.

5. The drug according to claim 3, characterized in that The dosage form of the medicine is an external cream.

6. An external cream capable of relieving pain symptoms of post-traumatic bone and joint degeneration, characterized in that: Calculated by weight, including:

7. The method for preparing the external-use cream according to claim 6, wherein The following steps are involved: Dissolve the emulsifier and glycerol in distilled water, add Atractylodes lactone I and mix thoroughly by ultrasonic dispersion to form a suspension, heat in a water bath to 50-90°C and incubate for 3-9 minutes to obtain an aqueous phase; Separately, take stearyl alcohol, white vaseline, and glyceryl monostearate and heat them on a water bath until they are completely dissolved and the temperature is between 50 and 90°C to obtain an oil phase; The oil phase is slowly added to the water phase while continuously stirring in the same direction until condensation occurs, thereby obtaining the external cream.

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