Smearing type composition percutaneously permeating into bone joint cavity and application of smearing type composition percutaneously permeating into bone joint cavity

Abstract

The invention discloses a smearing type composition percutaneously permeating into a bone joint cavity and application of the smearing type composition. The smearing type composition comprises the following raw materials in parts by weight: 75-90 parts of calcium-based meteorite ceramic powder (Ca-MCP), 2-5 parts of cortex dictamni, 2-5 parts of radix notoginseng, 1-3 parts of flos carthami, 3-8 parts of radix angelicae sinensis and 2-5 parts of radix angelicae pubescentis. Experimental results show that when the composition provided by the invention is directly smeared on an affected part, effective components in the composition can permeate into a bone joint cavity through skin, and the composition has good permeation capacity, is safe and convenient to use, and has a great application prospect in the field of bone disease treatment.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical preparations, and particularly relates to a composition for transdermal penetration into joint cavities by smearing and its application. Background Art

[0002] Knee osteoarthritis (OA) is a disease with local degenerative changes that occur as the knee joint cartilage wears away and disappears. It mainly affects the knee joint cartilage, subchondral bone, synovium, joint capsule and other structures of the joint, and is the leading cause of knee pain and disability in the elderly.

[0003] Currently, the conventional clinical treatment solutions for joint diseases are as follows: before the joint disease reaches the surgical indication conditions, doctors prescribe some oral or topical medications to relieve pain. When the disease severity reaches the surgical indication, joint replacement surgery is performed. This waiting process means that elderly patients have to endure bone pain. Moreover, even when the surgical indication is reached, due to the low immunity of elderly patients, they face high surgical risks and high recurrence risks after surgery. Therefore, only a very small number of elderly patients will choose surgical treatment for knee osteoarthritis, and most still use conservative treatment methods to relieve knee pain. So, the treatment of knee arthritis has been in a medical bottleneck.

[0004] Traditional topical medications for joint diseases cannot directly target and deliver drugs to the bone layer through skin penetration. Generally, they can only reach the skin layer. Even if some can reach the muscle layer, the drug components will be circulated to the internal body with the blood and cannot reach the bone layer. Therefore, studying transdermal drug delivery to reach the bone layer has become an important breakthrough in the treatment of knee osteoarthritis by transdermal drug delivery. Summary of the Invention

[0005] Aiming at the problems existing in the above-mentioned prior art, the purpose of the present invention is to provide a composition for transdermal penetration into joint cavities by smearing and its application.

[0006] To achieve the above technical purpose, the present invention provides the following technical solutions:

[0007] In the first aspect of the present invention, there is provided a composition for transdermal penetration into joint cavities by smearing. Calculated by 100 parts by weight in total, it includes the following raw materials in parts by weight: calcium-based meteorite ceramic powder (hereinafter abbreviated as Ca-MCP) 75 - 90 parts, Dictamnus dasycarpus Turcz. 2 - 5 parts, Panax notoginseng 2 - 5 parts, Carthamus tinctorius 1 - 3 parts, Angelica sinensis 3 - 8 parts, Heracleum hemsleyanum Diels 2 - 5 parts.

[0008] Preferably, the Ca-MCP is prepared with reference to a Chinese patent with the authorized publication number of CN 107586098B, and is processed by mixing chondrite, purple sand, and clay in a certain mass percentage. The mass percentages of chondrite, purple sand, and clay are 0.3-43%: 56-95%: 1-5%.

[0009] The second aspect of the present invention provides the application of the above-mentioned composition for transdermal penetration into the joint cavity by smearing in the preparation of drugs for treating bone diseases.

[0010] The present invention has the following beneficial effects:

[0011] The present invention provides a composition for transdermal penetration into the joint cavity by smearing, and its raw materials include calcium-based meteorite ceramic powder (Ca-MCP), Dictamnus dasycarpus Turcz., Panax notoginseng, Carthamus tinctorius, Angelica sinensis, and Heracleum hemsleyanum Diels. The experimental results show that when the composition provided by the present invention is directly smeared on the diseased part, it is safe and non-irritating to the skin, can penetrate through the skin into the joint cavity, and has a good penetration amount, and is safe and convenient to use. Clinical trials show that the composition provided by the present invention can be applied to the preparation of drugs for treating knee osteoarthritis. Therefore, it has a good application prospect. Detailed implementation manners

[0012] In the following description, specific details such as specific system structures and technologies are presented for the purpose of illustration rather than limitation, so as to thoroughly understand the embodiments of the present invention. However, those skilled in the art should clearly understand that the present invention can also be implemented in other embodiments without these specific details. In other cases, the detailed descriptions of well-known systems, devices, circuits, and methods are omitted to avoid unnecessary details from interfering with the description of the present invention.

[0013] Example 1

[0014] I. Animal experiments

[0015] 1. Experimental materials

[0016] 1.1 Instruments and consumables

[0017] CMA402 dual-channel microinjection pump, CMA130 in vitro fixing bracket, CMA470 micro-low-temperature step-by-step collector, 1.0mL glass microinjection syringe; R580 animal anesthesia machine and ThermoStar body temperature maintenance instrument, Forma 88000 series -80℃ ultra-low temperature refrigerator (Thermo Scientific, USA), microdialysis brain probe CMA 12Elite Metal Free (effective dialysis membrane length is 4mm, molecular retention capacity is 20kD), microdialysis linear probe CMA 30Linear MD Probe (effective dialysis membrane length is 10mm, molecular retention capacity is 6000D).

[0018] 1.2 Drugs and reagents

[0019] Ringer solution (Ringers) (batch number: A17022005, Hebei Tiancheng Pharmaceutical Co., Ltd.), isoflurane (batch number: 217180801, Shenzhen Ruiwode Life Science Co., Ltd.), and experimental water were prepared by Milli-Q Integral 5 ultrapure water machine, and methanol, formic acid and other reagents were all chromatographic grade.

[0020] Preparation of ointment composition:

[0021] (1) Preparation of Ca-MCP: Ca-MCP is prepared according to a method for producing meteorite ceramics with ultra-low heavy metal dissolution and low radioactivity disclosed in a Chinese patent application authorization announcement number: CN 107586098B.

[0022] S11. Material selection and grinding: prepare raw materials according to the following components: achondrite, purple sand, and clay; grind the achondrite into 200-mesh powder, grind the purple sand into 200-mesh powder, and grind the clay into 200-mesh powder;

[0023] S12, slurry preparation: dry the three raw materials in step S1, weigh 0.3g of achondrite, 95g of purple sand, and 4.7g of clay, mix them thoroughly, add 20g of water to prepare the slurry, and the Zn content in the slurry preparation water is more than 0.7mg / L, and the β radioactivity is less than 0.03Bq / L;

[0024] S13, kneading the clay: repeatedly kneading, beating and squeezing the mixture after slurry preparation in step S2 until the cross section of the clay is shiny;

[0025] S14, aging: the mixture obtained in step S3 is placed in a sealed stainless steel container and allowed to stand for 3 months;

[0026] S15, re-kneading the mud: kneading, beating, and squeezing the mixed material obtained in step S4 again to knead the mud;

[0027] S16. Shaping: Manually shape the mixture obtained in step S5.

[0028] S17. Drying: Air-dry the formed embryo in step S6.

[0029] S18. Baking: After loading the air-dried embryo in step S7 into the kiln, raise the temperature to 100 °C in 2 hours, hold for 2 hours, then raise the temperature to 200 °C in 40 minutes, hold at 200 °C for 1 hour, and take out the embryo for embryo trimming after natural cooling.

[0030] S19. Firing: Raise the temperature to 200 °C in 1 hour, then raise the temperature to 500 °C in 1.5 hours, hold for 20 minutes, then raise the temperature to 700 °C in 40 minutes and hold for 20 minutes for crystal transformation, then raise the temperature to 1160 °C in 1 hour and 40 minutes and hold for 0.5 hour, turn off the fire, open the air damper and cool naturally to 700 °C, close the air damper, open the air damper again when the temperature drops to 500 °C, and when the temperature drops below 160 °C, open the kiln door half-way for ventilation and cooling until the product is taken out of the kiln to obtain Ca-MCP.

[0031] (2) Weigh 82 g of Ca-MCP, 3 g of Dictamnus dasycarpus Turcz., 2.5 g of Panax notoginseng, 2.4 g of Carthamus tinctorius L., 4.5 g of Angelica pubescens Maxim., and 5.6 g of Angelica sinensis (Oliv.) Diels, mix them evenly to obtain an ointment composition.

[0032] 1.3 Experimental animals

[0033] 50 male Sprague-Dawley rats, weighing 250 - 280 g (SPF, specific pathogen-free), were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., license number: SCXK (Beijing) 2021 - 0006. They were housed in IVC cages (individually ventilated cages, length × width × height: 461 × 274 × 229 mm, Tecniplast, Italy), and the laboratory environmental temperature was maintained at 22 °C and the humidity was about 50%.

[0034] 2. Experimental methods

[0035] 2.1 Drug administration

[0036] The rats were depilated 1 day in advance. Before drug administration, the right thigh joint was hot compress for 3 min, and 1 g of the ointment composition was applied around the right thigh joint, and the drug was sealed and fixed with sealing film. The drug was administered once a day for 3 consecutive days.

[0037] 2.2 Implantation of microdialysis probes in rat joints and subcutaneous tissues

[0038] On the day of the microdialysis experiment (after 3 consecutive days of drug administration), rats were subjected to arthroscopic and subcutaneous microdialysis probe implantation experiments. After weighing the rats, they were anesthetized (isoflurane, 5.0% induction and 2% maintenance). A linear MD probe was used and inserted into the subcutaneous tissue by the accompanying guide needle using the "push-in - pull-out" method; for the joint cavity, a brain probe with a membrane length of 4 mm was used and inserted into the joint cavity through the bone pores on the front of the right posterior thigh knee joint guided by a puncture needle with a diameter of 1.5 mm.

[0039] 2.3 Drug administration to rats and collection of microdialysis samples

[0040] Apply 1 g of the ointment composition around the joints of the rats. Use a circular drug delivery patch to control the drug delivery area and prevent the ointment from spreading and contaminating the probe implantation holes. Immediately after drug administration, start collecting arthroscopic and subcutaneous microdialysis fluids. The microdialysis probe was perfused with compound sodium chloride at a rate of 1.5 μL·min -1 . The microdialysis fluid samples were collected in a micro low-temperature fraction collector (4 °C), once every 60 min, until 6 h after drug administration; then the microdialysis samples were transferred and stored in a -80 °C refrigerator for future measurement; after the dialysis was completed, the probes were removed, and plasma, joint bones, fascia, and soft tissues were taken and stored in a -80 °C refrigerator for future measurement. No adverse reactions were observed.

[0041] 1.3 HPLC-MS / MS quantitative method was used to detect the contents of the main active ingredients of the ointment composition in the microdialysis fluid, plasma, knee joint, and surrounding tissues of rats.

[0042] 1.3.1 Experimental drugs and instruments

[0043] The list of control drugs is shown in Table 1, and the remaining drugs are stored in the laboratory. Experimental water was prepared by a Milli-Q Integral 5 ultrapure water machine, and methanol, formic acid, and other reagents were all of chromatographic grade.

[0044] Table 1

[0045]

[0046] HPLC-MS / MS system (AB Sciex, USA), including ExionLC-20AC high-performance liquid chromatograph, IonDriveTM Turbo V ion source, Sciex 6500+ triple quadrupole detector, and Analyst 1.7 data acquisition system. Micro SB-5200D ultrasonic cleaner (domestic SCIENTZ).

[0047] 1.3.2 Detection method

[0048] (a) HPLC-MS / MS conditions

[0049] Chromatographic conditions: The chromatographic column was an XSelect@HSS T3 chromatographic column (2.5 μm, 2.1 mm × 50 mm, Waters, USA); the mobile phase was A (water, containing 0.05‰ formic acid)∶B (methanol, containing 0.05‰ formic acid), and the gradient elution program was as follows: 0 - 1.5 min, 15% B; 1.5 - 3.0 min, 15% B - 100% B; 3.0 - 4.5 min, 100% B; 4.50 - 4.51 min, 100% B - 15% B; 4.51 - 6.0 min, 15% B. The flow rate was 0.3 mL·min -1 ; the column temperature was 30°C; the sample chamber temperature was 46°C; the injection volume was 1 μL.

[0050] Mass spectrometry conditions: Electrospray ionization source (ESI), curtain gas (N2) was 40 psi (1 psi ≈ 6.9 kPa), collision gas (N2) was 9 psi, spray voltage was +5.5 kV, atomization temperature was 550°C, and both nebulizing gas (N2) and auxiliary gas (N2) were 55 psi. Positive ion scanning was performed in multiple reaction monitoring (MRM) mode, MRM (transition) was 325.1 / 109.0, declustering potential (DP) was 5, and collision energy (CE) was 35. The results are shown in Table 2.

[0051] Table 2

[0052]

[0053] As can be seen from the results in Table 2, the plaster composition contains the main plant medicinal materials Heracleum hemsleyanum, Dictamnus dasycarpus, Panax notoginseng, Angelica sinensis, and Carthamus tinctorius. Through literature retrieval for the treatment of joint-related diseases, with anti-inflammatory and other effects and combined with the index components in the pharmacopoeia, a total of 19 target components were selected for detection. The experimental results preliminarily showed that after administration of the plaster composition, three components, evodolide, dictamnine, and kaempferide, could be quantitatively detected in the knee joint cavity, subcutaneous tissue, and blood of rats. Among them, evodolide had the highest content in the osteoarticular microdialysate. The above results indicate that the composition prepared by the present invention can penetrate through the skin into the osteoarticular cavity.

[0054] II. Clinical experiment

[0055] In order to verify the therapeutic effect of the composition provided by the present invention on osteoarthritis, the following clinical experiment was provided.

[0056] 1. Knee osteoarthritis

[0057] (1) Research subjects: 30 patients with knee osteoarthritis were selected for the experiment at Beijing Mingyifang Traditional Chinese Medicine Hospital. The patients had symptoms such as pain during joint movement, joint tenderness, joint swelling, joint morning stiffness, difficulty in squatting and standing up, pain when going up and down stairs, and limited mobility. Among them, the patients' ages ranged from 40 to 77 years old, with an average age of 66.7 years, and the disease duration was 1 to 10 years, with an average of 3.46 years. The improvement of the VAS pain score of the patients was recorded before and after treatment.

[0058] (2) Usage method: The paste composition was hot compress applied to the affected knee, and a customized spectrum therapeutic instrument was used to heat at a temperature of 36° - 42° for 30 minutes, once every other day, and 12 times constituted a treatment course, with 10 g calculated according to the active ingredient each time.

[0059] (3) Efficacy criteria

[0060] Marked effect: The arthritis symptoms of the patient, such as joint pain, swelling, and limited mobility, were significantly reduced or completely disappeared; or the VAS pain score decreased by more than 60%;

[0061] Effective: The arthritis symptoms of the patient were improved; or the VAS pain score decreased by 30%;

[0062] Ineffective: The arthritis symptoms of the patient were not improved; or the VAS pain score did not decrease;

[0063] (4) Clinical experiment results

[0064] After all patients were continuously treated for 4 times - 2 treatment courses, the treatment results of the patients with knee osteoarthritis are shown in Table 3 below.

[0065] Table 3

[0066] Project Total number of people Number of people with marked efficacy Number of people with effective results Number of people with ineffective results Total marked efficacy and effective rate Total effective rate Knee osteoarthritis 30 29 30 0 96.6％ 100％

[0067] The above results indicate that the composition provided by this application is promising for use in the preparation of drugs for the treatment of knee osteoarthritis.

[0068] The present invention is not limited to the above specific embodiments. Those of ordinary skill in the art starting from the above concepts and making various transformations without creative labor fall within the protection scope of the present invention.

Claims

1. A composition for percutaneous penetration into joint cavities by smearing, characterized in that, It comprises the following raw materials in parts by weight: 75-90 parts of calcium-based meteorite ceramic powder, 2-5 parts of dictamnus bark, 2-5 parts of notoginseng, 1-3 parts of safflower, 3-8 parts of angelica, and 2-5 parts of pubescent angelica root.

2. The composition for percutaneous penetration into joint cavities by smearing according to claim 1, wherein The Ca-MCP is processed by mixing chondrite calcium meteorite, purple sand, and clay according to a certain mass percentage. The mass percentages of the chondrite calcium meteorite, purple sand, and clay are 0.3-43%: 56-95%: 1-5%.

3. Use of the composition for topical transdermal penetration into joint cavities according to claim 1 or 2 in the preparation of a medicament for treating bone diseases.

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