Use of a pharmaceutical composition in a transdermal drug delivery formulation

Abstract

The application discloses a kind of total glycosides of Paeonia lactiflora and non-steroidal anti-inflammatory drug composition in transdermal drug preparation application.In transdermal drug preparation the application advantage of this composition is in:(1) the active epidermal barrier and stratum corneum barrier of skin is the main obstacle of total glycosides of Paeonia lactiflora transdermal absorption.The non-steroidal anti-inflammatory drug in the composition can reversibly open the active epidermal barrier of skin, if it is used with the skin stratum corneum barrier of penetration enhancer and can significantly improve the transdermal penetration performance of total glycosides of Paeonia lactiflora in this transdermal drug preparation, it is a kind of with simple preparation process, easy industrial production, skin non-invasive characteristics can safely, efficiently improve the method for the transdermal penetration ability of total glycosides of Paeonia lactiflora;(2) total glycosides of Paeonia lactiflora and non-steroidal anti-inflammatory drug combination in transdermal drug preparation, in the curative effect of treating primary dysmenorrhea and rheumatoid arthritis disease has obvious synergistic effect, curative effect is excellent.

Description

Technical Field

[0001] This invention belongs to the field of transdermal drug delivery technology, specifically relating to the application of a pharmaceutical composition consisting of total glucosides of paeony and a nonsteroidal anti-inflammatory drug in transdermal drug delivery formulations. Background Technology

[0002] Dysmenorrhea is a common gynecological condition. Based on the presence or absence of organic lesions in the reproductive organs, it can be divided into primary dysmenorrhea (PD) and secondary dysmenorrhea. Primary dysmenorrhea, also known as functional dysmenorrhea, accounts for over 90% of dysmenorrhea cases. It refers to lower abdominal pain and distension experienced before, during, and after menstruation, without any organic lesions in the reproductive organs. It is often accompanied by headaches, fatigue, and lower back pain, severely impacting the patient's quality of life and work. The earliest record of dysmenorrhea is found in the *Synopsis of Prescriptions of the Golden Chamber* (Jin Gui Yao Lue), specifically in the section on "Miscellaneous Diseases of Women": "Leukorrhea, irregular menstruation, and fullness and pain in young women." Zhang Zhongjing pointed out that menstrual pain in women is closely related to poor blood flow. Traditional Chinese medicine considers primary dysmenorrhea to fall under the category of "menstrual abdominal pain." The root cause is attributed to poor blood and qi circulation, leading to "pain due to obstruction" or insufficient nourishment of the Chong and Ren meridians and the uterus. Therefore, the occurrence of primary dysmenorrhea is closely related to the cyclical physiological changes of the Chong and Ren meridians and the uterus. Improving the patient's menstrual blood deficiency and symptoms of insufficient nourishment of the Chong and Ren meridians and uterus during menstruation, or unblocking the flow of qi and blood in the Chong and Ren meridians and uterus during menstruation, are fundamental treatments for primary dysmenorrhea. Modern medicine, however, believes that endogenous substances in the endometrium, such as prostaglandin F2α (PGF2α), estrogen, progesterone, endothelin, nitric oxide, β-endorphin, calcium ions, and oxygen free radicals, can collectively promote the formation and development of primary dysmenorrhea through various pathways.

[0003] Traditional Chinese medicine commonly uses classic formulas for treating primary dysmenorrhea, such as Wenjing Decoction, Taohong Siwu Decoction, Aiqi Decoction, Shengyu Decoction, and Xuanyu Tongjing Decoction, all of which contain white peony root, indicating its effectiveness in treating primary dysmenorrhea. Traditional Chinese medicine believes that the liver and spleen are closely related to the generation and circulation of qi and blood. The liver stores blood and regulates blood flow; the spleen governs transformation and transportation, serving as the source of qi and blood production and possessing hematopoietic function. White peony root enters the liver and spleen meridians, excelling at nourishing blood and regulating menstruation. It nourishes the Chong and Ren meridians and the uterus, fundamentally improving the reduced uterine blood flow, uterine tissue ischemia, and hypoxia caused by "lack of nourishment," thus exerting its therapeutic effect on primary dysmenorrhea. Furthermore, qi stagnation leads to blood stasis, while qi circulation promotes blood circulation. White peony enters the liver meridian and promotes the free flow of qi. It can regulate the flow of qi, soothe the liver and relieve depression. By promoting blood flow and removing blood stasis, it can fundamentally treat primary dysmenorrhea caused by poor blood circulation and blood stasis in the uterus, which is characterized by "pain due to obstruction". Total glucosides of paeony (TGP) are the main active ingredients of the traditional Chinese medicine white peony root in treating primary dysmenorrhea. Modern pharmacological studies have shown that TGP can treat primary dysmenorrhea through multiple pathways: 1) Paeoniflorin can inhibit prothrombin production, inhibit platelet aggregation, dilate venous and peripheral small blood vessels, and improve blood microcirculation, thus improving dysmenorrhea symptoms caused by uterine ischemia and hypoxia through its effects of promoting blood circulation, replenishing blood, relieving spasms, and alleviating pain; 2) TGP can reduce the content of PGF2α in mouse uterine tissue, thereby inhibiting the spasmodic contraction of uterine smooth muscle and relieving primary dysmenorrhea symptoms; 3) TGP can increase the NO level in rat uterus, inhibit excessive uterine contraction, promote blood flow, and inhibit the production of anaerobic metabolites, thereby relieving primary dysmenorrhea symptoms; 4) TGP can inhibit extracellular calcium... 2+ 5) Paeoniflorin and paeoniflorin may exert analgesic effects by increasing serum and cerebral cortex β-EP levels, activating endogenous analgesic mechanisms, and inhibiting pain transmission and nociceptive excitation.

[0004] Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic synovitis and cartilage destruction, mainly manifested as polyarticular and symmetrical joint inflammation of small joints in the hands and feet. In severe cases, it can cause cartilage damage and joint destruction, leading to joint deformity and even loss of function, with strong disability-causing characteristics, seriously affecting the living conditions and work quality of patients. Traditional medicine believes that rheumatoid arthritis conforms to the common etiological factors of bi syndrome, belonging to the category of "bi syndrome" in traditional Chinese medicine, with the pathogenic characteristics of internal factors of liver and kidney deficiency and qi and blood deficiency, and external factors of wind, cold, and dampness pathogens. In patients with rheumatoid arthritis, due to their own qi and blood deficiency and yang damage, the腠理 (interstitial space) is empty and the exterior defense is not solid. At this time, wind, cold, and dampness pathogens are more likely to invade the skin, joints, and meridians, resulting in blocked qi and blood circulation, swelling and pain in the affected area, and joint deformation and stiffness. It can be seen that the occurrence of rheumatoid arthritis is closely related to the internal deficiency syndrome of the liver, kidney, and spleen organs, and the medication should be based on tonifying the liver and kidney, soothing the liver qi, and supplementing qi and activating blood circulation as the fundamental treatment. Modern medicine believes that various factors such as immune cells (synovial B cells, synovial macrophages, neutrophils, etc.), cytokines (IL-1, IL-6, IL-17, TNF-α, etc.), autoantigens, and intestinal flora may all cause the occurrence and development of rheumatoid arthritis.

[0005] White peony root is contained in the drug compositions of classic prescriptions for treating rheumatoid arthritis commonly used in traditional medicine, such as Shaoyao Gancao Decoction, Huangqi Guizhi Wuwu Decoction, and Guizhi Shaoyao Zhimu Decoction. It can be seen that it has good curative effects in the treatment of rheumatoid arthritis. Traditional Chinese medicine believes that if the liver qi is stagnated and the qi movement is不畅 (unsmooth), it can affect the qi and blood circulation and aggravate the bi syndrome. White peony root enters the liver meridian, and the liver governs the tendons. By nourishing the liver blood, it can play a role in moistening and nourishing the tendons, helping to relieve symptoms such as joint pain, contracture, and difficulty in flexion and extension. In addition, the liver is in charge of dredging and regulating qi. White peony root can soothe the liver qi, make the qi movement smooth, promote the qi and blood circulation, and contribute to the treatment of the bi syndrome. When white peony root enters the spleen meridian, it can strengthen the spleen and nourish the blood, making the generation of qi and blood have a source, providing a material basis for the repair of joints. Patients with rheumatoid arthritis often have unbearably painful joints, and the sour and cold nature of white peony root can astringe qi and blood, relieve spasm and pain, and has good therapeutic effects on symptoms such as joint pain and muscle spasm caused by the bi syndrome. Similarly, TGP is also the main active ingredient of white peony root in the treatment of rheumatoid arthritis. Modern pharmacological research shows that TGP can treat rheumatoid arthritis through multiple pathways: 1) exert the anti-inflammatory effect of TGP, by inhibiting the proliferation of fibroblast-like synoviocytes (FLS) and promoting the apoptosis of FLS cells, thereby reducing the release of inflammatory factors, playing an anti-inflammatory role and relieving the symptoms of rheumatoid arthritis; 2) exert the antithrombotic effect of TGP, making the blood circulation smooth, which helps to nourish the tendons and repair the damaged joints, thus achieving the effect of treating rheumatoid arthritis; 3) exert the immunomodulatory effect of TGP, by reducing the proportion of CD4+ cells in the spleen and lymph nodes, CD4+ Cells / CD8 + Cell ratio and IFN-γ level increase CD8 + 4) By regulating the ratio of cells, TGP enhances the phagocytic activity of macrophages against foreign cells, thereby regulating immune function, balancing the body's immune system, and improving rheumatoid arthritis symptoms; 5) By regulating the function of osteoclasts and osteoblasts, TGP inhibits the proliferation and differentiation of osteoclasts and protects the maturation and differentiation function of osteoblasts, thus playing a role in treating rheumatoid arthritis; 6) In patients with rheumatoid arthritis, TGP regulates the gut microbiota by significantly increasing the function of immune system-related microbiota and significantly decreasing the function of immune system disease-related microbiota, thereby inhibiting the occurrence of inflammatory responses and improving rheumatoid arthritis symptoms.

[0006] Although TGPs are effective in treating primary dysmenorrhea and rheumatoid arthritis symptoms, they also have significant drawbacks. Currently, the most common dosage form of TGPs is oral capsules, which have the following problems: 1) Due to the combined effects of the first-pass effect in the liver, the binding of P-glycoprotein to TGP, and the poor lipid solubility of TGP itself, oral absorption of TGPs is poor, resulting in a low absolute bioavailability of only 3%–4%; 2) The blood concentration of TGP capsules is unstable, and the drug has a short half-life and short duration of action, requiring multiple doses and leading to poor patient compliance; 3) The local blood concentration of oral TGPs is low, failing to produce a large enough concentration in the uterus or synovial membrane area to exert a local therapeutic effect. In contrast, transdermal TGP formulations offer advantages such as being free from the first-pass effect in the liver, higher bioavailability, a longer half-life, stable blood concentrations, and the ability to exert local therapeutic effects. However, the skin is a major barrier to the transdermal absorption of drugs into the body. Due to the high water solubility of TGP, it is difficult to achieve transdermal absorption. Although there are methods to improve the transdermal performance of TGP, they have the following problems: 1) Adding penetration enhancers such as propylene glycol, oleic acid, and berberine hydrochloride. The advantage of this method is that it can improve the transdermal performance of TGP to a certain extent, and the preparation process is relatively simple. However, the drawback is that although these penetration enhancers can effectively open the stratum corneum barrier of the skin, they cannot open the active epidermal barrier of the skin. The presence of the active epidermal barrier still limits the transdermal permeation of TGP. Therefore, the effect of using these penetration enhancers alone on improving the transdermal performance of TGP is very limited, and the penetration enhancement factor is usually only 1.5 to 2.5 times; 2) Using physical methods such as iontophoresis and microneedles. The advantage of this method is that it can improve the transdermal performance of TGP, but the drawback is that the preparation process is relatively complex, and the application of these invasive skin preparations also carries the risk of skin infection; 3) Using novel carriers such as liposomes, nanoparticles, and nanogels. While this type of method also improves the transdermal performance of TGP, its drawbacks include high production costs, demanding equipment requirements, and complex manufacturing processes. Therefore, given these issues, it is crucial to develop methods and strategies that can safely and efficiently enhance the transdermal permeability of TGP, ensuring that the preparation process is simple, easily industrialized, and non-invasive.

[0007] Nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly used in clinical practice to treat primary dysmenorrhea and rheumatoid arthritis, characterized by rapid onset of action. NSAIDs are antipyretic, analgesic, and anti-inflammatory drugs that do not contain steroidal structures; common examples include nimesulide, indomethacin, flufenamic acid, meloxicam, and aspirin. Modern pharmacological studies have shown that in treating primary dysmenorrhea, high concentrations of PGF2α can act on PGF2α receptors on the walls of spiral arterioles, leading to spasmodic contractions of uterine smooth muscle and a large accumulation of acidic products in the myometrium, thus triggering dysmenorrhea. Simultaneously, it increases the sensitivity of pelvic nerves to pain, amplifying the pain sensation. NSAIDs are highly effective prostaglandin synthase inhibitors; they can block the synthesis of prostaglandins by efficiently inhibiting cyclooxygenase, reducing PGF2α levels, thereby rapidly inhibiting spasmodic contractions of the endometrial smooth muscle and treating primary dysmenorrhea. In the treatment of rheumatoid arthritis, NSAIDs can inhibit cyclooxygenase activity, thereby inhibiting the synthesis of inflammatory mediators such as arachidonic acid and prostaglandins, including anterocycline and thromboxane A2. This can reduce inflammation, swelling, and pain in the synovial tissue, thus exerting a therapeutic effect on rheumatoid arthritis. However, although NSAIDs can rapidly relieve pain in primary dysmenorrhea and rheumatoid arthritis, their short half-life, short duration of action, frequent dosing, and high relapse rate after discontinuation limit their clinical application in these diseases. Summary of the Invention

[0008] Purpose of the invention: The purpose of this invention is to provide a pharmaceutical composition for use in transdermal drug delivery formulations, which can effectively improve the transdermal permeability of total glucosides of paeony, and at the same time produce a better synergistic therapeutic effect on primary dysmenorrhea and rheumatoid arthritis.

[0009] Technical solution: The pharmaceutical composition of the present invention comprises total glucosides of paeony and a nonsteroidal anti-inflammatory drug.

[0010] The active ingredients in the pharmaceutical composition are total paeoniflorin and nonsteroidal anti-inflammatory drugs.

[0011] The composition wherein the total paeoniflorin glycosides are paeoniflorin terpenoid components, including one or more of paeoniflorin, paeoniflorin lactone, oxypaeoniflorin, benzoylpaeoniflorin, and galloylpaeoniflorin, or extracts containing paeoniflorin terpenoid components extracted from traditional Chinese medicine or plants. Further, the traditional Chinese medicine or plants include Paeonia lactiflora and Paeonia suffruticosa.

[0012] The composition, wherein the nonsteroidal anti-inflammatory drug comprises one or more of diclofenac, indomethacin, flurbiprofen, flufenamic acid, nabumetone, meloxicam, rofecoxib, celecoxib, mefenamic acid, nimesulide, loxoprofen, etoricoxib, and pharmaceutically acceptable salts or derivatives thereof.

[0013] The use of the pharmaceutical composition in the preparation of drugs for treating primary dysmenorrhea and rheumatoid arthritis.

[0014] Application of nonsteroidal anti-inflammatory drugs in improving the transdermal properties of total glucosides of paeony.

[0015] Furthermore, the application of nonsteroidal anti-inflammatory drugs combined with total glucosides of paeony in transdermal drug delivery formulations.

[0016] The transdermal drug delivery formulation is prepared by adding pharmaceutically acceptable excipients to the pharmaceutical composition.

[0017] The transdermal drug delivery formulation, wherein the pharmaceutically acceptable excipients include chemical penetration enhancers. More preferably, the transdermal drug delivery formulation contains pharmaceutically acceptable chemical penetration enhancers for enhancing the ability of a drug to penetrate the stratum corneum barrier.

[0018] The transdermal drug delivery formulation wherein the chemical penetration enhancer includes one or more of pepper oil, peppermint oil, galangal oil, mustard oil, 1,8-cineole, evodia oil, dried ginger oil, and clove oil.

[0019] The transdermal drug delivery preparations described herein are selected from the following dosage forms: patches, emulsions, plasters, ointments, plasters, pastes, coatings, gels, powders, aerosols, sprays, oils, lotions, glycerin, liniments, suspensions, solutions, sols, tinctures, and medicated oils.

[0020] More preferably, when the transdermal drug delivery formulation is a patch, it contains 11-18 mg / cm³ of total paeoniflorin. 2 Contains 4-7 mg / cm³ of nonsteroidal anti-inflammatory drugs 2 .

[0021] In practice, the nonsteroidal anti-inflammatory drugs in the drug composition can enhance the transdermal permeability of total paeoniflorin.

[0022] In practice, the total glucosides of paeony in this drug composition have a significant synergistic effect with nonsteroidal anti-inflammatory drugs in the treatment of primary dysmenorrhea and rheumatoid arthritis.

[0023] Through long-term series of studies, the patent applicant discovered that the combined use of total paeoniflorin and nonsteroidal anti-inflammatory drugs in transdermal drug delivery formulations not only improves the bioavailability of the drug, prolongs the drug half-life, maintains a stable blood drug concentration, and exerts local therapeutic effects, but more importantly, the nonsteroidal anti-inflammatory drugs in the drug composition can significantly improve the transdermal permeability of total paeoniflorin. Moreover, the combination of the two has a significant synergistic effect in the treatment of primary dysmenorrhea and rheumatoid arthritis. The specific value analysis of the combination of nonsteroidal anti-inflammatory drugs and total paeoniflorin in this transdermal drug delivery formulation is as follows: (1) In terms of improving the transdermal permeability of total paeoniflorin, nonsteroidal anti-inflammatory drugs can reversibly open the tight junction of active epidermal tissue. When used in combination with a permeation enhancer that opens the stratum corneum barrier, the transdermal permeability of total paeoniflorin can be significantly enhanced. Therefore, the method of using nonsteroidal anti-inflammatory drugs (NSAIDs) to improve the transdermal permeability of total paeoniflorin can solve the problems existing in the known technologies. It has the advantages of effectively improving the transdermal permeability of total paeoniflorin, simple preparation process, no damage to the skin, and easy industrial production; (2) In terms of the efficacy of NSAIDs themselves, NSAIDs have a rapid onset of action and can quickly improve the symptoms of primary dysmenorrhea and rheumatoid arthritis. However, their drug half-life is relatively short, the duration of drug effect is relatively short, frequent use is required, and relapse is easy after drug withdrawal. These problems also lead to the limitations of NSAIDs in the clinical treatment of primary dysmenorrhea and rheumatoid arthritis. Compared with total paeoniflorin, although NSAIDs do not have the efficacy of sustained therapeutic effect of total paeoniflorin, they have the characteristic of rapid onset of action that total paeoniflorin does not have. Therefore, the combined use of nonsteroidal anti-inflammatory drugs and total glucosides of paeony has significant synergistic effects. The combination of the two can provide both rapid onset and sustained, long-lasting therapeutic benefits for primary dysmenorrhea and rheumatoid arthritis.

[0024] Beneficial Effects: Compared with the prior art, the present invention has the following advantages: (1) The transdermal drug delivery formulation made from the pharmaceutical composition provided by the present invention can significantly improve the transdermal permeability of total paeoniflorin. Compared with the existing methods for enhancing the transdermal permeability of total paeoniflorin, applying this composition to the transdermal drug delivery formulation is a safe and efficient method to improve the transdermal permeability of total paeoniflorin, which has the characteristics of simple preparation process, easy industrial production, and non-invasive skin treatment. (2) The combined use of this pharmaceutical composition in the transdermal drug delivery formulation provided by the present invention has a significant synergistic effect in the treatment of primary dysmenorrhea and rheumatoid arthritis, with excellent efficacy. Attached Figure Description

[0025] Figure 1 The graph shows the permeation enhancement effect of flufenamic acid on paeoniflorin.

[0026] Figure 2The graph shows the permeation enhancement effect of celecoxib on paeoniflorin.

[0027] Figure 3 The graph shows the permeation enhancement effect of flurbiprofen on paeoniflorin.

[0028] Figure 4 The image shown is a graph illustrating the permeation enhancement effect of nimesulide on paeoniflorin. Detailed Implementation

[0029] The present invention will be further described below with reference to specific embodiments. It should be understood that these embodiments are illustrative and are only used to illustrate the present invention, and are not intended to limit the scope of the invention. The pharmaceutical compositions, composition patches, and contrast patches provided in the following specific embodiments were all prepared in the laboratory. Any imitation, modification, or alteration made without departing from the pharmaceutical combination concept and type provided by the present invention is within the protection scope of the present invention. The raw materials and equipment used in the present invention are all known products and were obtained by purchasing commercially available products.

[0030] Example 1

[0031] This embodiment provides a pharmaceutical composition consisting of flufenamic acid and paeoniflorin, named Composition 1; the weight ratio of flufenamic acid to paeoniflorin is 1:1, and the mixture is prepared in the laboratory.

[0032] Example 2

[0033] This embodiment provides a pharmaceutical composition consisting of celecoxib and paeoniflorin, named composition 2; the weight ratio of celecoxib to paeoniflorin is 1:1, and the mixture is prepared in the laboratory.

[0034] Example 3

[0035] This embodiment provides a pharmaceutical composition consisting of flurbiprofen and paeoniflorin, named composition 3; the weight ratio of flurbiprofen to paeoniflorin is 1:1, and the mixture is prepared in the laboratory.

[0036] Example 4

[0037] This embodiment provides a pharmaceutical composition consisting of nimesulide and paeoniflorin, named composition 4; the weight ratio of nimesulide to paeoniflorin is 1:1, and the mixture is prepared in the laboratory.

[0038] Example 5

[0039] This embodiment provides a transdermal drug delivery patch, composed of composition 5 (paeoniflorin and mefenamic acid) and other pharmaceutical excipients, named Example 5 Patch, which was prepared in the laboratory. The specific preparation method is as follows:

[0040] 5.1 Prescription Preparation

[0041] All components are in parts by weight:

[0042]

[0043] Among them, paeoniflorin is the total glycoside of paeoniflorin; mefenamic acid is a non-steroidal anti-inflammatory drug component; hydroxypropyl cellulose is used as a skeleton material; triethyl citrate is used as a plasticizer; butyl acrylate-acrylic acid copolymer is used as a binder; polyvinylpyrrolidone is used as a humectant; magnesium aluminum silicate is used as a crosslinking agent; ascorbic acid is used as an antioxidant and crosslinking regulator; and clove oil is used as a stratum corneum penetration enhancer.

[0044] 5.2 Preparation method

[0045] (1) Paeoniflorin, mefenamic acid, hydroxypropyl cellulose, triethyl citrate and an appropriate amount of water phase are fully swollen and uniformly dispersed to obtain mixture a;

[0046] (2) Mix butyl acrylate-acrylic acid copolymer, polyvinylpyrrolidone, clove oil and an appropriate amount of water to obtain mixture b;

[0047] (3) Mix magnesium aluminum silicate, ascorbic acid and the remaining aqueous phase to obtain mixture c;

[0048] (4) Combine mixtures a, b, and c and stir thoroughly. Place the resulting total mixture between the coating machine lining (non-woven fabric) and the protective layer (ethylene-vinyl acetate copolymer) and extrude to obtain a patch containing composition 5.

[0049] (5) The obtained patches were placed in a dryer and baked for 30 minutes. The finished product had a moisture content of approximately 7.4%. After removal, the patches were cut into 3cm × 3cm pieces and packaged. HPLC analysis revealed a moisture content of 18.5 mg / cm³. 2 The patch of composition 5 (containing 12.58 mg / cm³ of paeoniflorin) 2 Contains mefenamic acid 5.92 mg / cm³ 2 ).

[0050] Example 6

[0051] This embodiment provides a transdermal drug delivery patch, composed of composition 6 (benzoylpaeoniflorin, galloylpaeoniflorin, diclofenac) and other pharmaceutical excipients, named Example 6 patch, which was prepared in the laboratory. The specific preparation method is as follows:

[0052] 6.1 Prescription Preparation

[0053] All components are in parts by weight:

[0054]

[0055] Among them, benzoylpaeoniflorin and galloylpaeoniflorin are the total glycosides of paeony; diclofenac is a non-steroidal anti-inflammatory drug component; polyacrylate is used as a skeleton material; triethyl citrate is used as a plasticizer; hydroxypropyl methylcellulose is used as a binder; sodium hyaluronate is used as a moisturizer; magnesium aluminum silicate is used as a crosslinking agent; lipoic acid is used as an antioxidant; ethylenediaminetetraacetic acid is used as a crosslinking regulator; and galangal oil is used as a stratum corneum penetration enhancer.

[0056] 6.2 Preparation method

[0057] (1) Benzoyl paeoniflorin, galloyl paeoniflorin, diclofenac, polyacrylate, triethyl citrate and an appropriate amount of water phase are fully swollen and uniformly dispersed to obtain mixture I;

[0058] (2) Hydroxypropyl methylcellulose, sodium hyaluronate, lipoic acid, galangal oil and an appropriate amount of water are mixed evenly to obtain mixture II;

[0059] (3) Mix magnesium aluminum silicate, ethylenediaminetetraacetic acid and the remaining aqueous phase to obtain mixture III;

[0060] (4) Combine mixtures I, II and III and stir them thoroughly. Place the resulting total mixture between the coating machine lining (non-woven fabric) and the protective layer (aluminum foil-polyethylene composite film) and extrude to obtain a patch containing composition 6.

[0061] (5) The obtained patches were placed in a dryer and baked for 30 minutes. The finished product had a moisture content of approximately 8.5%. After removal, the patches were cut into 3cm × 3cm pieces and packaged. HPLC analysis revealed a moisture content of 22.5 mg / cm³. 2 Composition 6 patch (containing benzoyl paeoniflorin 11.25 mg / cm³) 2 Contains 5.25 mg / cm³ of galloylpaeoniflorin. 2 Contains diclofenac 6.00 mg / cm³ 2 ).

[0062] Comparative Example 1

[0063] This comparative example provides a patch, named Comparative Example 1 Patch. The only difference between Comparative Example 1 Patch and the patch of Example 5 is that Comparative Example 1 Patch does not contain paeoniflorin, and the reduced mass is proportionally allocated to the mass of mefenamic acid, while other components and contents remain unchanged.

[0064] The preparation method is described in Example 5.

[0065] Comparative Example 2

[0066] This comparative example provides a patch, named Comparative Example 2 Patch. The only difference between Comparative Example 2 Patch and the patch of Example 5 is that Comparative Example 2 Patch does not contain mefenamic acid, and its reduced mass is proportionally allocated to the mass of paeoniflorin, while other components and contents remain unchanged.

[0067] The preparation method is described in Example 5.

[0068] Comparative Example 3

[0069] This comparative example provides a patch, named Comparative Example 3 Patch. The only difference between Comparative Example 3 Patch and the patch of Example 5 is that the Comparative Example 3 Patch is a base patch that does not contain components of Composition 5 (paeoniflorin and mefenamic acid), and the reduced mass of these two components is proportionally allocated to the mass of the other components.

[0070] The preparation method is described in Example 5.

[0071] Comparative Example 4

[0072] This comparative example provides a patch, named Comparative Example 4 Patch. The only difference between Comparative Example 4 Patch and the patch of Example 6 is that it does not contain benzoylpaeoniflorin and galloylpaeoniflorin, and the reduced mass of these two compounds is proportionally allocated to the mass of diclofenac, while other components and their contents remain unchanged.

[0073] The preparation method is described in Example 6.

[0074] Comparative Example 5

[0075] This comparative example provides a patch, named Comparative Example 5 Patch. The only difference between Comparative Example 5 Patch and the patch of Example 6 is that Comparative Example 5 Patch does not contain diclofenac, and its reduced mass is proportionally allocated to the mass of benzoylpaeoniflorin and galloylpaeoniflorin, while other components and contents remain unchanged.

[0076] The preparation method is described in Example 6.

[0077] Comparative Example 6

[0078] This comparative example provides a patch, named Comparative Example 6 Patch. The only difference between Comparative Example 6 Patch and Example 6 Patch is that Comparative Example 6 Patch is a base patch that does not contain components of Composition 6 (benzoylpaeoniflorin, galloylpaeoniflorin, and diclofenac), and the reduced mass of these three components is proportionally allocated to the mass of the other components.

[0079] The preparation method is described in Example 6.

[0080] Experimental Example 1. Effect of flufenamic acid on the transdermal properties of paeoniflorin

[0081] 1.1 Main experimental reagents and instruments

[0082] Drugs and reagents: paeoniflorin, composition 1, peppermint oil, urethane, phosphoric acid, acetonitrile, physiological saline, anhydrous ethanol.

[0083] Instruments: Transdermal diffusion tester, high performance liquid chromatograph, high speed centrifuge.

[0084] 1.2 Laboratory Animals

[0085] Five female, common-grade New Zealand rabbits, weighing 2-3 kg each.

[0086] 1.3 Analytical Methods

[0087] 1.3.1 High Performance Liquid Chromatography Conditions

[0088] Chromatographic column: Luna C18 column (150*4.6mm, 5μm); mobile phase: acetonitrile: 0.1% phosphoric acid aqueous solution (14:68, V / V); flow rate: 1.0mL / min; detection wavelength: 230nm; temperature: 30℃; injection volume: 20μL.

[0089] 1.3.2 Preparation of standard solutions and establishment of standard curves

[0090] Accurately weigh paeoniflorin reference standard, add ethanol to dilute to volume, and shake well to obtain the standard solution. Perform linear regression with paeoniflorin mass concentration as the x-axis and peak area as the y-axis.

[0091] 1.4 Rabbit skin in vitro transdermal diffusion experiment method

[0092] 1.4.1 Preparation of isolated rabbit skin

[0093] After anesthetizing the rabbits with urethane solution, the rabbits' abdominal hair was removed using a shaver. Each rabbit was then euthanized, and the abdominal skin was cut off, washed with physiological saline, and stored in an ice bath for later use.

[0094] 1.4.2 Transdermal test

[0095] Skin samples from each rabbit were randomly divided into two groups: a control group and an experimental group. A Franz diffusion cell was used with PBS as the receiving medium. The control group received OP-10 solution containing 5% paeoniflorin (containing 7% peppermint oil), while the experimental group received OP-10 solution containing 5% paeoniflorin (combination 1, paeoniflorin + flufenamic acid, containing 7% peppermint oil). Skin samples from each group were placed in a transdermal diffusion apparatus and subjected to continuous stirring at 32°C and 600 rpm. 0.5 mL samples were taken from the receiving cell at 1, 2, 4, 5, and 6 hours, and an equal volume of fresh receiving medium at the same temperature was immediately added.

[0096] 1.4.3 Data Processing

[0097] The above samples were placed in a high-speed centrifuge and centrifuged at 12000 r·min⁻¹. 1 Centrifuge for 10 min under the specified conditions. Determine the paeoniflorin content in each group of samples under the chromatographic conditions described in section "1.3.1" and calculate the cumulative transdermal drug per unit area (Q) at different time points. n ).

[0098] 1.5 Experimental Results and Analysis

[0099] The results are shown in Table 1. Figure 1 As shown in the figure, compared with the control group, the experimental group showed a significant increase in the cumulative transdermal penetration of paeoniflorin (P<0.001). These results indicate that the combined use of flufenamic acid and peppermint oil significantly enhances the transdermal penetration of paeoniflorin compared to peppermint oil alone. This is because while peppermint oil effectively opens the stratum corneum barrier, it cannot open the active epidermal barrier, whereas flufenamic acid can open the active epidermal barrier. Therefore, the combined use of flufenamic acid and peppermint oil significantly enhances the transdermal properties of paeoniflorin.

[0100] Table 1. Results of paeoniflorin permeation enhancement ( n=5)

[0101]

[0102] Note: Compared with the control group, * P<0.05, ** P < 0.01, *** P<0.001, **** P < 0.0001.

[0103] Experimental Example 2. Effect of celecoxib on the transdermal properties of paeoniflorin

[0104] 2.1 Main experimental reagents and instruments:

[0105] Replace Composition 1 under “1.1” with Composition 2, replace peppermint oil with galangal oil, and use the same experimental reagents and instruments as in Experiment 1.

[0106] 2.2 Experimental animals, analytical methods, and preparation of isolated rabbit skin

[0107] Same as Experiment 1.

[0108] 2.3 Rabbit skin in vitro transdermal diffusion experiment method

[0109] 2.3.1 Transdermal test

[0110] The skin of each rabbit was randomly divided into two groups: a control group and an experimental group. A Franz diffusion cell was used with PBS as the receiving medium. The control group was supplied with an OP-10 solution containing 5% paeoniflorin (containing 7% galangal oil); the experimental group was supplied with an OP-10 solution containing 5% paeoniflorin (combination 2, paeoniflorin + celecoxib) (containing 7% galangal oil). The transdermal assay method and procedure were the same as in Example 1.

[0111] 2.3.2 Data Processing

[0112] The data processing method is the same as in Experiment 1.

[0113] 2.4 Experimental Results and Analysis

[0114] The results are shown in Table 2. Figure 2 As shown in the figure, compared with the control group, the experimental group showed a significant increase in the cumulative transdermal penetration of paeoniflorin (P<0.001). These results indicate that the combined use of celecoxib and galangal oil significantly enhances the transdermal penetration of paeoniflorin compared to galangal oil alone. This is because while galangal oil effectively opens the stratum corneum barrier, it cannot open the active epidermal barrier, whereas celecoxib can open the active epidermal barrier. Therefore, the combined use of celecoxib and galangal oil significantly enhances the transdermal performance of paeoniflorin.

[0115] Table 2. Results of paeoniflorin permeation enhancement ( n=5)

[0116]

[0117] Note: Compared with the control group, * P<0.05, ** P < 0.01, *** P<0.001, **** P < 0.0001.

[0118] Experimental Example 3. Effect of Flurbiprofen on the Transdermal Properties of Paeoniflorin

[0119] 3.1 Main experimental reagents and instruments:

[0120] Replace paeoniflorin under item “1.1” with paeoniflorin lactone, replace composition 1 with composition 3, replace peppermint oil with 1,8-cineole, and use the same experimental reagents and instruments as in Experiment 1.

[0121] 3.2 Laboratory Animals

[0122] Same as Experiment 1.

[0123] 3.3 Analytical Methods

[0124] 3.3.1 High Performance Liquid Chromatography Conditions

[0125] Chromatographic column: Luna C18 column (150*4.6mm, 5μm); mobile phase: acetonitrile: 0.1% phosphoric acid aqueous solution (13:87, V / V); flow rate: 1.0mL / min; detection wavelength: 233nm; temperature: 30℃; injection volume: 20μL.

[0126] 3.3.2 Preparation of standard solutions and establishment of standard curves

[0127] Weigh out paeoniflorin, add ethanol to make up to volume, and shake well to obtain a standard solution. Perform linear regression with the mass concentration of paeoniflorin as the x-axis and the peak area as the y-axis.

[0128] 3.4 In vitro transdermal diffusion experiment method using rabbit skin

[0129] 3.4.1 Preparation of isolated rabbit skin

[0130] The preparation method for isolated rabbit skin is the same as in Experiment 1.

[0131] 3.4.2 Transdermal test

[0132] The skin of each rabbit was randomly divided into two groups: a control group and an experimental group. A Franz diffusion cell was used with PBS as the receiving medium. The control group was supplied with an OP-10 solution containing 5% paeoniflorin (containing 7% 1,8-cineole); the experimental group was supplied with an OP-10 solution containing 5% paeoniflorin, composition 3 (paeoniflorin + flurbiprofen) (containing 7% 1,8-cineole). The transdermal assay method and procedure were the same as in Example 1.

[0133] 3.4.3 Data Processing

[0134] The samples from each group were placed in a high-speed centrifuge and centrifuged at 12000 r·min⁻¹ for 10 min. The content of paeoniflorin in each group of samples was determined according to the chromatographic conditions under section “3.3.1”, and the cumulative transdermal drug per unit area (Q') at different time points was calculated. n ).

[0135] 3.5 Experimental Results and Analysis

[0136] The results are shown in Table 3. Figure 3As shown in the figure, compared with the control group, the experimental group showed a significant increase in the cumulative transdermal penetration of paeoniflorin (P<0.001). These results indicate that the combined use of flurbiprofen and 1,8-cineole significantly enhances the transdermal permeability of paeoniflorin compared to 1,8-cineole alone. This is because while 1,8-cineole effectively opens the stratum corneum barrier, it fails to open the active epidermal barrier, whereas flurbiprofen can open the active epidermal barrier. Therefore, the combined use of flurbiprofen and 1,8-cineole significantly enhances the transdermal performance of paeoniflorin.

[0137] Table 3. Results of permeation enhancement by paeoniflorin ( n=5)

[0138]

[0139] Note: Compared with the control group, * P<0.05, ** P < 0.01, *** P<0.001, **** P < 0.0001.

[0140] Experimental Example 4. Effect of Nimesulide on the transdermal properties of paeoniflorin

[0141] 4.1 Main experimental reagents and instruments:

[0142] Replace composition 3 under section “3.1” with composition 4, and replace 1,8-cineole with Evodia rutaecarpa oil. Other experimental reagents, instruments and equipment are the same as in test example 3.

[0143] 4.2 Experimental animals, analytical methods, and preparation of isolated rabbit skin

[0144] Same as Experiment 1.

[0145] 4.3 In vitro transdermal diffusion experiment method using rabbit skin

[0146] 4.3.1 Transdermal test

[0147] The skin of each rabbit was randomly divided into two groups: a control group and an experimental group. A Franz diffusion cell was used with PBS as the receiving medium. The control group was supplied with an OP-10 solution containing 5% paeoniflorin (containing 7% Evodia rutaecarpa oil); the experimental group was supplied with an OP-10 solution containing 5% paeoniflorin, composition 4 (paaeoniflorin + nimesulide) (containing 7% Evodia rutaecarpa oil). The transdermal assay method and procedure were the same as in Example 1.

[0148] 4.3.2 Data Processing

[0149] Same as Experiment 1.

[0150] 4.4 Experimental Results and Analysis

[0151] The results are shown in Table 4. Figure 4 As shown in the figure, compared with the control group, the experimental group showed a significant increase in the cumulative transdermal penetration of paeoniflorin (P<0.001). These results indicate that the combined use of nimesulide and Evodia rutaecarpa oil significantly enhances the transdermal penetration of paeoniflorin compared to using Evodia rutaecarpa oil alone. This is because while Evodia rutaecarpa oil effectively opens the stratum corneum barrier, it cannot open the active epidermal barrier, whereas nimesulide can open the active epidermal barrier. Therefore, the combined use of nimesulide and Evodia rutaecarpa oil significantly enhances the transdermal properties of paeoniflorin.

[0152] Table 4. Results of permeation enhancement of paeoniflorin ( n=5)

[0153]

[0154] Note: Compared with the control group, * P<0.05, ** P < 0.01, *** P<0.001, **** P < 0.0001.

[0155] Experimental Example 5. Evaluation of the efficacy of the patches from Example 5 and Example 6 in treating primary dysmenorrhea in mice.

[0156] 5.1 Experimental reagents and materials

[0157] Loxoprofen sodium tablets, total glucosides of paeony tablets, patch of Example 5, patch of Comparative Example 1, patch of Comparative Example 2, patch of Comparative Example 3, patch of Example 6, patch of Comparative Example 4, patch of Comparative Example 5, patch of Comparative Example 6, estradiol benzoate injection, oxytocin injection.

[0158] 5.2 Laboratory Animals

[0159] 120 female KM mice, weighing 18–22g.

[0160] 5.3 Grouping and Dosing

[0161] One hundred and twenty female KM mice (n=10 per group) were randomly divided into four groups: normal group, model group, loxoprofen sodium group (100 mg / kg), total paeoniflorin group (100 mg / kg), patch group of Example 5, patch group of Comparative Example 1, patch group of Comparative Example 2, patch group of Comparative Example 3, patch group of Example 6, patch group of Comparative Example 4, patch group of Comparative Example 5, and patch group of Comparative Example 6. After acclimatization for four days, the loxoprofen sodium group and the total paeoniflorin group were given the corresponding dose of drug, while the normal group and the model group were given the same dose of distilled water. Two patches of the corresponding drug were applied to the abdomen of each patch group, and the drug was administered once daily for seven consecutive days. Starting from day 5, except for the normal group mice which were subcutaneously injected with saline, the other groups of mice were simultaneously injected subcutaneously with estradiol benzoate injection (0.3 mg / kg) daily. One hour after the last administration on day 7 (the corresponding patches were not removed after the last administration in each patch group), oxytocin (20 U / kg) was injected intraperitoneally. The mice were considered to have successfully established the model when they showed a writhing response with abdominal retraction, hind limb extension, and buttock elevation.

[0162] 5.4 Detection Indicators

[0163] Mice were placed in an incubator immediately after oxytocin injection. The number of writhing responses was observed and recorded at 0–1 h, 3–4 h, and 6–7 h after oxytocin injection. The inhibition rate of the number of writhing responses was calculated.

[0164] Inhibition rate of writhing response count in mice (%) = (Mean number of writhing responses in the model group - Mean number of writhing responses in each treatment group) / Mean number of writhing responses in the model group × 100%

[0165] 5.5 Experimental Results and Conclusions

[0166] (1) The writhing behavior of mice within 0-1 hour after oxytocin injection is shown in Table 5. Compared with the normal group, the number of writhing responses in the model group mice was significantly increased (P<0.0001), indicating successful modeling. Compared with the model group, the number of writhing responses in mice in the loxoprofen sodium group, total paeoniflorin group, patch group of Example 5, patch group of Comparative Example 1, patch group of Comparative Example 2, patch group of Example 6, patch group of Comparative Example 4, and patch group of Comparative Example 5 was significantly decreased (P<0.0001, P<0.05, P<0.0001, P<0.0001, P<0.05, P<0.0001, P<0.0001, P<0.05), while the number of writhing responses in the patch group of Comparative Example 3 and patch group of Comparative Example 6 was similar and showed no significant difference (P>0.05).

[0167] (2) The writhing behavior of mice within 3-4 hours after oxytocin injection is shown in Table 6. Compared with the normal group, the number of writhing responses in the model group mice was significantly increased (P<0.0001), indicating successful modeling. Compared with the model group, the number of writhing responses in mice in the total glucosides of paeony group, the patch group of Example 5, the patch group of Comparative Example 1, the patch group of Comparative Example 2, the patch group of Example 6, the patch group of Comparative Example 4, and the patch group of Comparative Example 5 was significantly decreased (P<0.001, P<0.0001, P<0.05, P<0.01, P<0.0001, P<0.05, P<0.01), while the number of writhing responses in the patch groups of Comparative Example 3 and Comparative Example 6 was similar, with no significant difference (P>0.05).

[0168] (3) The writhing behavior of mice within 6-7 hours after oxytocin injection is shown in Table 7. Compared with the normal group, the number of writhing responses in the model group mice was significantly increased (P<0.0001), indicating successful modeling. Compared with the model group, the number of writhing responses in mice in the total paeoniflorin group, the patch group of Example 5, the patch group of Comparative Example 2, the patch group of Example 6, and the patch group of Comparative Example 5 was significantly decreased (P<0.0001, P<0.0001, P<0.05, P<0.0001, P<0.05), while the number of writhing responses in the patch groups of Comparative Example 3 and Comparative Example 6 was similar, with no significant difference (P>0.05).

[0169] The above results indicate that both the patches in Example 5 and Example 6 effectively alleviated writhing symptoms in mice with primary dysmenorrhea (the matrix in the patches had no drug effect), and their efficacy was superior to that of the nonsteroidal anti-inflammatory drug (NSAID) comparative patch, the total paeoniflorin comparative patch, or the NSAID and total paeoniflorin administered by gavage. The NSAID comparative patch or the NSAID administered by gavage showed significant effects in alleviating writhing symptoms in the dysmenorrhea model mice in the early stages, but the effect was short-lived and quickly weakened. While the total paeoniflorin administered by gavage showed similar effects in alleviating writhing symptoms in the dysmenorrhea model mice in the later stages of treatment to the two combined patches, its therapeutic effect in the early stages was significantly inferior to the combined patches. Due to the active epidermal barrier, the total paeoniflorin comparative patch had a limited permeable amount, thus only exerting a weak effect, and its efficacy remained limited after administration. The two aforementioned patch compositions incorporate nonsteroidal anti-inflammatory drugs (NSAIDs) that reversibly open the active epidermal barrier. This not only significantly increases the transdermal permeability of total paeoniflorin but also endows the patch composition with the rapid onset of action characteristic of NSAIDs. Therefore, the patch composition exhibits the best therapeutic effect, demonstrating rapid, sustained, and long-lasting efficacy in treating primary dysmenorrhea in mice.

[0170] Table 5. Statistical results of the number of writhing responses and the inhibition rate of writhing responses in each group of mice within 0-1 hour. n=10)

[0171]

[0172] Note: Compared with the model group, * P<0.05, ** P < 0.01, *** P<0.001, **** P < 0.0001; compared with the normal group, ### P<0.001, #### P<0.0001.

[0173] Table 6. Statistical results of the number of writhing responses and the inhibition rate of writhing responses in each group of mice within 3-4 hours. n=10)

[0174]

[0175]

[0176] Table notes: Same as table notes in Table 5.

[0177] Table 7. Statistical results of the number of writhing responses and the inhibition rate of writhing responses in each group of mice within 5-6 hours. n=10)

[0178]

[0179] Table notes: Same as table notes in Table 5.

[0180] Experimental Example 6. Evaluation of the efficacy of the patches from Example 5 and Example 6 in treating adjuvant-induced arthritis in rats.

[0181] 6.1 Experimental reagents and materials

[0182] Nimesulide tablets, total glucosides of paeony tablets, patch of Example 5, patch of Comparative Example 1, patch of Comparative Example 2, patch of Comparative Example 3, patch of Example 6, patch of Comparative Example 4, patch of Comparative Example 5, patch of Comparative Example 6, Freund's complete adjuvant.

[0183] 6.2 Laboratory Animals

[0184] Seventy-two male SD rats, weighing 150–180 g.

[0185] 6.3 Grouping, Modeling, and Drug Administration

[0186] Seventy-two male rats, six in each group, were randomly divided into the following groups: normal group, model group, nimesulide group (150 mg / kg), total glucosides of paeony group (150 mg / kg), patch group of Example 5, patch group of Comparative Example 1, patch group of Comparative Example 2, patch group of Comparative Example 3, patch group of Example 6, patch group of Comparative Example 4, patch group of Comparative Example 5, and patch group of Comparative Example 6. After one week of acclimatization, rats in all groups (except the normal group) were injected intradermally with 0.1 mL of complete Freund's adjuvant per rat in their right hind paw to induce inflammation. On the second day, prophylactic administration was performed: the normal group and the model group were given the same dose of distilled water; the nimesulide group and the total glucosides of paeony group were given the corresponding dose of drug by gavage; and each patch group had half of the corresponding patch applied to the right hind paw of the rat, and was administered continuously for 3 weeks (once a day). One hour after the second administration at the end of the fourth week, 0.1 mL of complete Freund's adjuvant per rat was injected again to induce inflammation (the corresponding patch was not removed after the last administration in each patch group).

[0187] 6.4 Detection Indicators

[0188] The number of times rats in each group licked their right hind paw at 0–0.5 h, 3–3.5 h, and 6–6.5 h after stimulation of inflammation was observed and recorded.

[0189] 6.5 Experimental Results and Conclusions

[0190] (1) The results of the number of times the right hind paw licked by rats within 0-0.5 h after stimulation of inflammation in each group are shown in Table 8: Compared with the normal group, the number of times the model group rats licked their paws was significantly increased (P<0.0001), indicating that the model was successfully established. Compared with the model group, the number of times the rats in the nimesulide group, total glucosides of paeony group, patch group of Example 5, patch group of Comparative Example 1, patch group of Comparative Example 2, patch group of Example 6, patch group of Comparative Example 4, and patch group of Comparative Example 5 were significantly decreased (P<0.0001, P<0.05, P<0.0001, P<0.0001, P<0.05, P<0.0001, P<0.0001, P<0.05), while the number of times the rats in the patch group of Comparative Example 3 and patch group of Comparative Example 6 were similar and there was no significant difference (P>0.05).

[0191] (2) The results of the number of times the rats licked their right hind paws within 3-3.5 hours after the inflammation was induced in each group are shown in Table 8. Compared with the normal group, the number of times the rats in the model group licked their paws was significantly increased (P<0.0001), indicating that the model was successfully established. Compared with the model group, the number of times the rats in the total glucosides of paeony group, the patch group of Example 5, the patch group of Comparative Example 1, the patch group of Comparative Example 2, the patch group of Example 6, the patch group of Comparative Example 4, and the patch group of Comparative Example 5 were significantly decreased (P<0.001, P<0.0001, P<0.05, P<0.01, P<0.0001, P<0.05, P<0.01), while the number of times the rats in the patch group of Comparative Example 3 and the patch group of Comparative Example 6 were similar to those in the model group, with no significant difference (P>0.05).

[0192] (3) The results of the number of times the rats licked their right hind paws within 6-6.5 h after the stimulation of inflammation are shown in Table 8. Compared with the normal group, the number of times the rats in the model group licked their paws was significantly increased (P<0.0001), indicating that the model was successfully established. Compared with the model group, the number of times the rats in the total glucosides of paeony group, the patch group of Example 5, the patch group of Comparative Example 2, the patch group of Example 6, and the patch group of Comparative Example 5 were significantly decreased (P<0.0001, P<0.0001, P<0.05, P<0.0001, P<0.05), while the number of times the rats in the patch group of Comparative Example 3 and the patch group of Comparative Example 6 were similar and there was no significant difference (P>0.05).

[0193] The above results indicate that both the patch in Example 5 and the patch in Example 6 can effectively alleviate the paw-licking symptoms in rats with adjuvant arthritis (the matrix in the patch has no drug effect), and their efficacy is significantly better than that of the patch alone using nonsteroidal anti-inflammatory drugs, the patch with total glucosides of paeony, or the patch administered by gavage with nonsteroidal anti-inflammatory drugs and total glucosides of paeony. The nonsteroidal anti-inflammatory drug (NSAID) contrast patch or NSAID gavage administration showed significant efficacy in relieving paw-licking symptoms in rats with adjuvant arthritis in the early stages, but the effect was short-lived and quickly diminished. While gavage administration of total paeoniflorin showed similar efficacy to the two aforementioned combination patches in relieving paw-licking symptoms in the later stages, its therapeutic effect in the early stages was significantly weaker. The total paeoniflorin contrast patch, due to the active epidermal barrier, had limited transdermal penetration, thus exerting only a weak effect and its efficacy remained limited after administration. In contrast, the aforementioned two combination patches incorporated NSAIDs with reversible epidermal barrier-opening properties, significantly increasing the transdermal penetration of total paeoniflorin and giving the combination patch the rapid onset of action characteristic of NSAIDs. Therefore, the combination patch offered the best therapeutic effect, providing a rapid, sustained, and long-lasting therapeutic advantage in rats with adjuvant arthritis.

[0194] Table 8. Statistics on the number of times rats licked their paws in each group ( n=6)

[0195]

[0196] Table notes: Same as table notes in Table 5.

[0197] Test Example 7. Skin irritation evaluation of patches from Example 5 and Example 6

[0198] 7.1 Main Drugs and Reagents

[0199] Example 5 patch, Example 6 patch, Comparative Example 3 patch, Comparative Example 6 patch, physiological saline.

[0200] 7.2 Laboratory Animals

[0201] 20 New Zealand rabbits of standard grade, half male and half female, weighing 2-3 kg.

[0202] 7.3 Skin irritation test

[0203] 7.3.1 Hair removal treatment for domestic rabbits

[0204] Rabbits were dehaired on the abdomen 24 hours before the experiment. Before administering the drug, the skin was observed for any damage. The rabbits with damaged skin should not be subjected to the irritation experiment.

[0205] 7.3.2 Grouping and Dosing

[0206] The 20 New Zealand rabbits were randomly divided into 4 groups, half male and half female, with 5 rabbits in each group. The rabbits were treated according to the following groups: (1) Applying the patch of Example 5 to the left side of the hairless area on the rabbit's abdomen; (2) Applying the patch of Comparative Example 3 to the right side of the hairless area on the rabbit's abdomen at the same frequency and dosage; (3) Applying the patch of Example 6 to the left side of the hairless area on the rabbit's abdomen; (4) Applying the patch of Comparative Example 6 to the right side of the hairless area on the rabbit's abdomen at the same frequency and dosage. Each rabbit was treated with 3 patches once daily for 7 consecutive days. One day after the last treatment, the patches were gently and slowly removed, the medication was washed off with warm water, and the rabbits were observed immediately (day 0). Erythema and swelling of the rabbit's skin were observed and recorded on days 1, 3, 5, and 7.

[0207] 7.4 Evaluation Indicators

[0208] 7.4.1 Skin Irritation Evaluation Criteria

[0209] Skin irritation is evaluated using erythema and edema as indicators, and the scoring criteria are as follows:

[0210] (1) Erythema: 0 points for no erythema; 1 point for barely visible mild erythema; 2 points for clearly visible moderate erythema; 3 points for severe erythema; and 4 points for purplish-red erythema to mild eschar formation.

[0211] (2) Edema: No edema is 0 points; mild edema that is barely visible is 1 point; moderate edema with obvious swelling is 2 points; severe edema with skin swelling of 1 mm and clear outline is 3 points; severe edema with skin swelling of more than 1 mm or with blisters or ulceration is 4 points.

[0212] The highest skin irritation score for each rabbit is 8 points, consisting of erythema and edema.

[0213] 7.4.2 Evaluation criteria for skin irritation intensity

[0214] The mean skin irritation intensity of each group of rabbits = (total score of skin erythema + edema in each group of rabbits) / number of rabbits in each group. The irritation intensity scoring criteria are as follows:

[0215] A mean value of 0–0.49 is considered non-irritating; 0.5–2.99 is considered mildly irritating; 3.0–5.99 is considered moderately irritating; and 6.0–8.00 is considered severely irritating.

[0216] 7.5 Results and Analysis of Skin Irritation Tests

[0217] The results are shown in Tables 9 and 10. The skin irritation intensity evaluation of both the patch from Example 5 and the patch from Example 6 in rabbits showed no irritation. These results indicate that neither the patch from Example 5 nor the patch from Example 6 causes skin irritation when applied topically.

[0218] Table 9. Skin irritation rating of the patch in Example 5 on rabbits (n=5)

[0219]

[0220] Table 10. Skin irritation rating of the patch in Example 6 on rabbits (n=5)

[0221]

Claims

1. A transdermal drug delivery formulation, characterized in that, It is prepared from total paeoniflorin, nonsteroidal anti-inflammatory drugs, chemical penetration enhancers and pharmaceutically acceptable excipients; the total paeoniflorin is selected from paeoniflorin, paeoniflorin lactone, benzoylpaeoniflorin, galloylpaeoniflorin; the nonsteroidal anti-inflammatory drugs are selected from flufenamic acid, celecoxib, flurbiprofen, nimesulide, mefenamic acid, diclofenac; the chemical penetration enhancers are selected from peppermint oil, galangal oil, 1,8-cineole, evodia oil, clove oil.

2. The transdermal drug delivery formulation according to claim 1, characterized in that, The dosage form of the preparation is selected from patches, emulsions, plasters, ointments, plasters, pastes, coatings, gels, powders, aerosols, sprays, oils, lotions, glycerin, liniments, suspensions, solutions, sols, tinctures, and medicated oils.

3. The use of the transdermal drug delivery formulation according to claim 1 in the preparation of a medicament for treating primary dysmenorrhea and rheumatoid arthritis.

4. Application of nonsteroidal anti-inflammatory drugs in the preparation of transdermal penetration enhancers that improve the transdermal permeability of total paeoniflorin, wherein the nonsteroidal anti-inflammatory drugs are selected from flufenamic acid, celecoxib, flurbiprofen, nimesulide, mefenamic acid, and diclofenac, and the total paeoniflorin is selected from paeoniflorin, paeoniflorin lactone, benzoylpaeoniflorin, and galloylpaeoniflorin.

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