A gel patch for treating lower back pain caused by intervertebral disc degeneration

Abstract

The present application relates to a kind of gel patch for treating lower back pain caused by intervertebral disc degeneration, which is composed of backing layer, drug storage layer and anti-adhesion layer, wherein the drug storage layer is composed of active ingredients, matrix and penetration enhancer; characterized in that the active ingredients are volatile oil and flow extract prepared from raw materials in the following weight parts: turmeric 17.8-21.7%, angelica 11.8-14.5%, frankincense 11.8-14.5%, myrrh 11.8-14.5%, salvia miltiorrhiza 11.8-14.5%, rhizoma drynariae 8.3-10.1%, morindae 8.3-10.1%, eucommia 8.3-10.1%. The present gel patch has a remarkable effect on treating lower back pain caused by intervertebral disc degeneration.

Description

TECHNICAL FIELD

[0001] The present application relates to the medicine preparation of traditional herbal medicine with uncertain structure, in particular to the medicine for treating bone disease mainly made of Curcuma longa L. BACKGROUND

[0002] Intervertebral disc degeneration (IDD) is one of the main causes of low back pain (LBP) in clinical. With the aggravation of population aging in China, LBP has become a common clinical symptom, and its incidence is gradually increasing. More than 80% of the intervertebral discs (IVD) in people over 50 years old show degeneration-related changes. At present, the treatment of IDD by western medicine is divided into surgical treatment and non-surgical treatment. According to statistics, the recurrence rate of postoperative pain of lumbar vertebrae can reach 5.0%-74.6%, and the rate of secondary surgery can reach 13.4%-35.0%, and the surgical treatment is expensive. In addition, long-term follow-up found that some patients after surgery had different degrees of lumbar instability, adjacent segment degeneration and other complications. Non-surgical drug treatment mainly includes non-steroidal anti-inflammatory drugs, cartilage protectors, etc. These drugs have certain effect on temporary relief of symptoms, but long-term use of such drugs has large side effects, damages the gastrointestinal mucosal barrier, and increases the incidence of gastrointestinal ulcers, gastric cancer and other diseases.

[0003] In contrast, traditional Chinese medicine treatment of IDD has the advantages of small side effects, low price, definite curative effect, and treatment of both symptoms and root causes. At present, in clinical practice, the main traditional Chinese medicines for treating lower back pain caused by intervertebral disc degeneration are listed traditional Chinese medicines and prescription drugs. Among them, the listed traditional Chinese medicines mainly include: Chanma Zhengtong Tincture (see National Standard Compilation of Chinese Patent Drugs, Orthopedics Volume, page 13), Henggu Gushangguhuan Agent (see Chinese Pharmacopoeia 2020, Volume 1, page 1356), Jingyaokang Capsules (see National Standard Compilation of Chinese Patent Drugs, Orthopedics Volume, page 335), Wudizhitong Liniment (see the 19th volume of Traditional Chinese Medicine Formulas of the National Health Commission of the People's Republic of China), Yaobitong Capsules (see Chinese Pharmacopoeia 2020, Volume 1, page 1805), and Yaotongning Capsules (Chinese Pharmacopoeia 2020, Volume 1, page 1802); for intervertebral disc degeneration causing lower back pain, although the prescriptions used by different doctors are not the same, most of them are composed of the following three types of drugs: (1) wind-dampness dispelling drugs, (2) blood-activating drugs, and (3) traditional Chinese medicinal materials related to the disease (Bai J. Research Progress of Traditional Chinese Medicine Treatment of Lumbar Intervertebral Disc Degeneration [J]. Liaoning Journal of Traditional Chinese Medicine, 2021, 48(09): 210-212. DOI: 10.13192 / j.issn.1000-1719.2021.09.059.). Looking at the above-mentioned listed traditional Chinese medicines and prescription drugs, their main effects are to relieve the symptoms of lumbar pain caused by blockage of meridians.

[0004] Another study by scholars using the classical prescription Huoluo Xielingdan to treat blood stasis type of lumbar intervertebral disc herniation [such as: (Qian Tianyu. Huoluo Xielingdan in the Treatment of 88 Cases of Blood Stasis Type of Lumbar Intervertebral Disc Herniation [J]. Zhejiang Journal of Traditional Chinese Medicine, 2016, 51(06): 441. DOI: 10.13633 / j.cnki.zjtcm.2016.06.036) (Liu Qiming. Clinical Efficacy of Huoluo Xielingdan on Patients with Lumbar Intervertebral Disc Herniation [J]. World Latest Medical Information Digest, 2016, 16(02): 136.)] proved that the classical prescription Huoluo Xielingdan has a certain effect on lumbar intervertebral disc herniation in intervertebral disc degeneration. As we all know, Huoluo Xielingdan is derived from the book Medical Insights from Chinese and Western Medicine written by Zhang Xichun, in which Mr. Zhang said, “This prescription has a strong ability to circulate and dissolve blood, and is effective in treating internal and external ulcers, abdominal and limb pain, and diseases caused by blood stasis.” This prescription is good at activating blood and resolving stasis, and is effective in treating various pains caused by blood stasis, whether it is an internal medicine or a surgical ulcer. However, the research results of the present inventors show that the classical prescription Huoluo Xielingdan is still not very effective in treating lower back pain caused by intervertebral disc degeneration (not simply lower back pain caused by lumbar intervertebral disc herniation).

[0005] In addition, the existing various prescription drugs are all oral decoctions, which not only have the disadvantage of first-pass effect, but also need to pass through the liver and intestine circulation to reach the disease site, and the curative effect is obviously impaired.

[0006] Topical medications, as preparations absorbed through the skin, not only reduce the side effects of drugs but also combine rapid efficacy, comfort, and simple administration. According to Traditional Chinese Medicine (TCM) classification, lower back pain can be categorized into Qi and Blood Stasis type, Damp-Heat and Phlegm Stagnation type, Wind-Cold-Dampness Bi syndrome type, and Liver and Kidney Deficiency type. Holistic examination and syndrome differentiation are the most fundamental characteristics of TCM treatment for degenerative diseases.

[0007] Huoluo Xiaoling Dan is indicated for: Qi and blood stagnation syndrome. Symptoms include abdominal pain, leg and arm pain, bruises and swelling from falls, internal and external sores, and masses and accumulations.

[0008] The symptoms treated by this formula are all caused by blood stasis and qi stagnation, and obstruction of the meridians. Therefore, the treatment should focus on promoting blood circulation, removing blood stasis, and relieving pain by unblocking the meridians. When the meridians are unobstructed and blood and qi flow smoothly, the stagnation will dissipate, swelling will subside, and pain in the heart, abdomen, legs, and arms will be relieved. Angelica sinensis, in this formula, is pungent, sweet, and warm, acting as a qi-regulating herb in the blood. It reduces swelling and relieves pain, nourishes and promotes blood circulation, and resolves stasis without harming the body's vital energy. Combined with Salvia miltiorrhiza, it enhances the power of promoting blood circulation and removing blood stasis. Myrrh and frankincense are both excellent herbs for promoting blood circulation and relieving pain. When used together, they help to promote blood circulation and remove blood stasis, further enhancing the effect of relieving pain and reducing swelling and nodules. The four herbs used together have the effect of promoting blood circulation, unblocking the meridians, and relieving pain. As the original text states: "This formula, in promoting the flow of qi and blood, has a great power to melt qi and blood, treating internal and external sores, pain in the heart, abdomen, and limbs. Most diseases caused by qi and blood stagnation are rare." Summary of the Invention

[0009] The technical problem to be solved by the present invention is to provide a gel patch for treating lower back pain caused by intervertebral disc degeneration, which has a significant effect on treating lower back pain caused by intervertebral disc degeneration.

[0010] The technical solution of the present invention to solve the above problems is as follows:

[0011] A gel patch for treating lower back pain caused by intervertebral disc degeneration, the gel patch comprising a backing layer, a drug-reservoir layer, and an anti-adhesion layer, wherein the drug-reservoir layer comprises an active ingredient, a matrix, and a penetration enhancer; characterized in that the active ingredient is a volatile oil and fluid extract made from the following raw materials in parts by weight: turmeric 17.8–21.7%, angelica 11.8–14.5%, frankincense 11.8–14.5%, myrrh 11.8–14.5%, salvia miltiorrhiza 11.8–14.5%, dipsacus asper 8.3–10.1%, mulberry mistletoe 8.3–10.1%, and eucommia ulmoides 8.3–10.1%.

[0012] In the above technical solution, the optimal ratio of the raw materials is: 19.6% turmeric, 13.2% angelica sinensis, 13.2% frankincense, 13.2% myrrh, 13.2% salvia miltiorrhiza, 9.2% dipsacus asper, 9.2% mulberry mistletoe, and 9.2% eucommia ulmoides.

[0013] In the technical scheme, the penetration enhancer is capsaicin; the matrix is a common gel patch matrix, which is specifically composed of sodium polyacrylate (high molecular skeleton material), citric acid (crosslinking regulator), glycerol (humectant), polyvinyl alcohol (viscosity increasing agent), gelatin (excipient) and aluminum glycinate (crosslinking agent); the backing layer is non-woven fabric or polyester film; and the protective layer is anti-sticking treated paper or polyester film.

[0014] In the technical scheme, the effective component is prepared by the following method:

[0015] The raw material is extracted by water vapor distillation twice. The first time, 8 times the amount of water is used, and the water vapor distillation is extracted for 2 hours, and the volatile oil and the medicinal liquid are collected respectively. The residue is added with 6 times the amount of water, and the second time of water vapor distillation is extracted for 1 hour, and the volatile oil and the medicinal liquid are collected respectively. The medicinal liquid extracted by water vapor distillation twice is combined and rotary evaporated by a rotary evaporator to a flow extract corresponding to a crude drug concentration of 2 g / mL. The obtained flow extract and the volatile oil extracted by water vapor distillation twice are the effective component.

[0016] In the technical scheme, the storage layer is composed of 46.63% of the effective component, 53.26% of the matrix and 0.11% of the penetration enhancer by weight percentage.

[0017] In the technical scheme, the preparation method of the paste of the storage layer is as follows:

[0018] 1. 0.15% of polyvinyl alcohol by weight of the flow extract is taken, and then added into 150 times the amount of water in a 90°C water bath to swell, followed by adding the flow extract and stirring uniformly. Then, 25% of gelatin by weight of the flow extract is added to swell to obtain phase A.

[0019] 2. Citric acid of 10.63% by weight of the volatile oil and aluminum glycinate of 5.31% by weight of the volatile oil are dissolved in water, and then the capsaicin is dissolved in ethanol. Then, the two are combined and added with the volatile oil and glycerol of 75% by weight of the volatile oil, and stirred uniformly to obtain phase B.

[0020] 3. In a 50°C water bath, phase A prepared in step 1 is mixed with phase B prepared in step 2 uniformly to obtain a mixed solution.

[0021] 4. 14.86% of sodium polyacrylate by weight of the effective component is added to the mixed solution prepared in step 3, and water bath is continued at 50°C to mix uniformly to obtain the paste of the storage layer.

[0022] As can be seen from the preparation method of the paste of the storage layer, the carboxylate ions (COO-) in the sodium polyacrylate molecules and the aluminum ions (Al 3+The hydrogel, formed by cross-linking between the molecules through electrostatic interactions to create a three-dimensional network structure, is easy to mold and is flexible and viscous. Meanwhile, capsaicin is used as a penetration enhancer, generating a constant temperature (feeling temperature 35–42°C) that lasts for up to 24 hours, increasing local blood circulation, clearing the meridians, and thus accelerating the improvement of the condition.

[0023] The gel patch described in this application is prepared by the following common methods:

[0024] The ointment of the drug storage layer prepared according to the above method is evenly applied to the non-woven fabric used as the backing layer, and then covered with an anti-adhesive layer (a polyester protective film treated with anti-adhesive), rolled into shape, and the thickness is controlled to be less than 5mm; finally, after sterilization, it is cut according to specifications, quality inspected, and packaged to obtain the gel patch.

[0025] The active ingredients in the gel patch of this invention are derived from eight herbs: turmeric, angelica, frankincense, myrrh, salvia miltiorrhiza, dipsacus root, eucommia bark, and mulberry mistletoe. In this formula, turmeric is pungent, bitter, and warm in nature, promoting blood circulation, regulating qi, and relieving pain, primarily treating lower back pain due to qi stagnation and blood stasis; therefore, it is the principal herb. Angelica, salvia miltiorrhiza, frankincense, and myrrh are combined as assistant herbs. The combined use of frankincense and myrrh disperses blood stasis, promotes qi circulation, and relieves pain, enhancing the blood-activating, qi-regulating, and pain-relieving effects of the principal herb, turmeric. The combination of Angelica sinensis and Salvia miltiorrhiza invigorates and replenishes blood, assisting Curcuma longa in invigorating blood without causing stagnation, and resolving stagnation without harming the body's vital energy. Eucommia ulmoides, Dipsacus asper, and Loranthus parasiticus work synergistically to tonify the liver and kidneys, strengthen tendons and bones, and benefit blood and unblock meridians. These three herbs act as adjuvants, assisting the assistant herbs in promoting qi and blood circulation and strengthening the body's foundation. Eucommia ulmoides primarily targets the lower body's qi aspect, excelling at tonifying kidney qi; Dipsacus asper and Loranthus parasiticus primarily target the lower body's blood aspect, dispelling wind in the blood, nourishing liver and kidney yin, and restoring deficiency. This combination is particularly suitable for chronic arthralgia, kidney deficiency, and lower back pain. The combined effects of these herbs, working synergistically, invigorate blood and unblock meridians, tonify the liver and kidneys, nourish yin and blood, and warm yang to strengthen the lower back, thus achieving the therapeutic effect of treating intervertebral disc degeneration.

[0026] The prescription of the effective component in the gel patch is improved on the basis of the traditional Chinese medicine prescription Jifang Huoluo Xiaoling Dan, and the improvement is that: from the medicinal taste, turmeric, eucommia ulmoides, rhizoma dangshen and parasitic four medicines are added; 2, from the prescription structure, the added turmeric is used as the monarch drug, and the four medicines in the original prescription are combined as the minister drug. The above improvement can produce the following effects: turmeric is pungent, bitter, warm, and belongs to the spleen and liver meridians, can break blood and move qi, dredge the channels and stop pain, and is used to treat blood stasis and qi stagnation; at the same time, the four medicines of angelica sinensis, salvia miltiorrhiza, frankincense and myrrh in the original prescription can activate blood and remove stasis, dredge the collaterals and stop pain, and enhance the function of breaking blood, moving qi and stopping pain of the monarch drug turmeric, so as to solve the symptoms of swelling and numbness of the lumbar collaterals caused by qi stagnation; on the other hand, angelica sinensis can tonify blood to counteract the excessive pungent and bitter tastes of turmeric, frankincense and myrrh, so as to prevent the damage of blood and stomach, and the four medicines can help the monarch drug turmeric to activate blood without stasis, and to remove stasis without damaging the normal, so as to solve the lumbar stabbing pain caused by blood collaterals not being nourished; in addition to the monarch drug turmeric, the newly added eucommia ulmoides, rhizoma dangshen and taxillus can be used to treat lower qi and can nourish liver and kidney yin to recover deficiency, and the three medicines are used together to regulate qi and blood, which is beneficial to tonifying kidney and strengthening waist to solve the root of lumbar pain, and is suitable for people with kidney deficiency and lumbar soreness caused by long-term arthralgia; secondly, the newly added eucommia ulmoides, rhizoma dangshen and taxillus are long in tonifying kidney, benefiting qi, strengthening bones and muscles, and tonifying blood and dredging vessels, which can help the minister drug to activate blood and dredge collaterals while tonifying kidney, strengthening waist and consolidating the body.

[0027] From the above analysis, it can be seen that the treatment method of the prescription of the application is changed from activating blood, removing stasis, dredging collaterals and stopping pain to breaking blood, moving qi, dredging channels and stopping pain, and at the same time, the important medicine for tonifying kidney and strengthening waist is added, which greatly enhances the effect of treating lower back pain caused by intervertebral disc degeneration.

[0028] In addition, capsaicin is a compound that makes chili taste very hot, so it is not only a commonly used penetration enhancer, but also a compound that can stimulate the skin to rapidly warm up. According to the research of the present inventors, the specific selection of capsaicin as a penetration enhancer can produce a constant body temperature of 35-42℃ in the waist of the patient, so as to increase local blood circulation, dredge the meridians, and significantly improve the curative effect.

[0029] In order for the public to better understand the present application, the beneficial effects of the present application are further illustrated by experiments and specific embodiments. BRIEF DESCRIPTION OF DRAWINGS

[0030] Figure 1 To establish a rat model of lumbar pain caused by intervertebral disc degeneration, code A in the figure represents the intervertebral disc annulus, and P represents puncture.

[0031] Figure 2 The arrows in the figure represent the L4-5 intervertebral disc.

[0032] Figure 3The arrow in the figure indicates the L4-5 intervertebral disc. DETAILED DESCRIPTION

[0033] Example 1 (preparation of gel patch)

[0034] Example 1

[0035] 1. Preparation of prescription of active ingredients:

[0036] Turmeric 75g, Angelica 50g, Frankincense 50g, Myrrh 50g, Salvia 50g, Dipsacus 35g, Taxillus 35g, Eucommia 35g.

[0037] 2. Preparation method of active ingredients:

[0038] According to the prescription, the above-mentioned 8 raw medicinal materials are taken by the amount, and steam distillation extraction is carried out twice. The first time is 8 times the amount of water, and the water vapor distillation extraction is carried out for 2 hours, and the volatile oil and the medicinal liquid are collected respectively. The residue is added with 6 times the amount of water, and the second time of water vapor distillation extraction is carried out for 1 hour, and the volatile oil and the medicinal liquid are collected respectively. The medicinal liquid of the two times of water vapor distillation extraction is combined, and rotary evaporation is carried out by a rotary evaporator to 200g of flow extract corresponding to the crude drug concentration of 2g / mL. The volatile oil of the two times of water vapor distillation extraction is combined to obtain 1.88g of volatile oil.

[0039] 3. Preparation of ointment of drug storage layer of gel patch:

[0040] (1) Ointment formula of drug storage layer:

[0041] Active ingredients: 200g of flow extract and 1.88g of volatile oil are prepared in this example; formula of matrix: 30g of sodium polyacrylate, 0.2g of citric acid, 150g of glycerol, 0.3g of polyvinyl alcohol, 50g of gelatin, 0.1g of aluminum glycinate; penetration enhancer: 0.5g of capsaicin.

[0042] (2) Preparation of ointment of drug storage layer:

[0043] (2.1) According to the formula of the matrix, polyvinyl alcohol is taken and first added with 150 times the amount of water in a 90°C water bath kettle for swelling, then the flow extract is added and stirred uniformly, and then the gelatin is added and sealed for swelling to obtain phase A;

[0044] (2.2) According to the formula of the matrix, citric acid and aluminum glycinate are dissolved in water, and then the capsaicin is dissolved in ethanol, and then the two are combined and added with volatile oil and glycerol, and stirred uniformly to obtain phase B;

[0045] (2.3) In a 50°C water bath kettle, the A phase prepared in step (2.1) and the B phase prepared in step (2.2) are mixed uniformly to obtain a mixed solution;

[0046] (2.4) Add sodium polyacrylate into the mixed solution prepared in step (2.3), continue water bath in 50°C water bath and mix evenly, to obtain the paste of the drug storage layer.

[0047] 4. Preparation of the gel patch

[0048] The paste of the drug storage layer prepared by the above method is evenly coated on the non-woven fabric as the backing layer, and then a polyester protective film treated by anti-sticking is overlaid as the anti-sticking layer, and then rolled and formed with the thickness controlled below 5mm. Finally, the gel patch 200 is prepared by sterilization, punching according to the specifications, quality inspection and packaging.

[0049] Example 2

[0050] 1. Prescription of the effective ingredients: turmeric 82.5g, angelica 52.5g, frankincense 50.3g, myrrh 47.6g, danshen 45.1g, huangqi 33.5g, sangjisheng 33.5g, and duzhong 35.0g.

[0051] 2. Preparation method of the effective ingredients: the volatile oil 1.98g and the flow extract 197.6g are prepared by the same method as in Example 1.

[0052] 3. Preparation of the paste of the drug storage layer of the gel patch

[0053] (1) Paste formula of the drug storage layer:

[0054] Effective ingredients: the flow extract 197.6g and the volatile oil 1.98g are prepared in this example; matrix formula: sodium polyacrylate 29.66g, citric acid 0.21g, glycerol 149.69g, polyvinyl alcohol 0.296g, gelatin 49.4g, and aluminum glycinate 0.11g; penetration enhancer: capsaicin 0.49g.

[0055] (2) Preparation of the paste of the drug storage layer:

[0056] In this example, the method for preparing the paste of the drug storage layer is the same as in Example 1.

[0057] 4. Preparation of the gel patch

[0058] In this example, the method for preparing the gel patch is the same as in Example 1, and the gel patch 197 is prepared.

[0059] Example 3

[0060] 1. Prescription of the effective ingredients: turmeric 67.5g, angelica 47.1g, frankincense 49.7g, myrrh 51.9g, danshen 54.6g, huangqi 38.0g, sangjisheng 36.5g, and duzhong 34.7g.

[0061] 2. Preparation of the effective component: The volatile oil 1.80 g and the flow extract 203 g were prepared by the same method as in Example 1.

[0062] 3. Preparation of the paste of the storage layer of the gel patch

[0063] (1) Paste formula of the storage layer

[0064] Effective component: The flow extract 203.0 g and the volatile oil 1.80 g were prepared in this example; matrix formula: sodium polyacrylate 30.43 g, citric acid 0.19 g, glycerol 152.25 g, polyvinyl alcohol 0.304 g, gelatin 50.75 g, and aluminum glycinate 0.09 g; penetration enhancer: capsaicin 0.51 g.

[0065] (2) Preparation of the paste of the storage layer

[0066] In this example, the method for preparing the paste of the storage layer was the same as in Example 1.

[0067] 4. Preparation of the gel patch

[0068] In this example, the method for preparing the gel patch was the same as in Example 1, and the gel patch 203 was prepared.

[0069] Example 2: (treatment effect research example)

[0070] I. Drug efficacy experiment

[0071] 1. Experimental method

[0072] 1.1. Experimental animals

[0073] 42 SD healthy rats, clean grade, 6-8 weeks old, weighing 250 g ± 20 g, male.

[0074] ① Main experimental equipment

[0075] TD-Ⅱ fully automatic thin-layer plate laying machine (Shanghai Kezhe Biochemical Technology Co., Ltd.); WGL-125B electric heating drying oven (Tianjin Tester Instrument Co., Ltd.); BSA224S-CW electronic balance (Sartorius Scientific Instruments Beijing Co., Ltd.); rotary evaporator (Shanghai Shensheng Technology Co., Ltd.); TC-15 nested thermostat (Haining Xinhua Medical Instrument Factory); TNF-α (Sinopharm Chemical Reagent Co., Ltd.); CFA (Shanghai Maclean Reagent Co., Ltd.); TNF-α detection kit (item number EK382 / 3, Lianke Biotechnology); IL-1β (item number SEA563Ra, Youersheng Biotechnology); ZH-YLS-6B intelligent hot plate apparatus (Anhui Zhenghua Bio-instrument Equipment Co., Ltd.); 7.0T Agilent MRI scanner (Agilent Technologies, USA). Micro-injector; 21g lumbar puncture needle; experimental medicinal materials were purchased from the Traditional Chinese Medicine Pharmacy of Nanfang Hospital Affiliated to Southern Medical University.

[0076] ② Test drug

[0077] Positive drug group: Wantong Muscle and Bone Plaster was purchased from Guangdong CR Shunfeng Pharmaceutical Co., Ltd.;

[0078] Experimental group: the gel patch of Example 1 above;

[0079] Control group 1: The four ingredients of Angelica sinensis, Salvia miltiorrhiza, Boswellia carterii and Commiphora myrrha were removed from the prescription of the effective ingredients in Example 1 above, and then the gel patch was prepared according to the method in Example 1 above.

[0080] Control group 2: The four ingredients of turmeric, eucommia, dipsacus and mulberry mistletoe were removed from the prescription of the effective ingredients in Example 1 above, and then the gel patch was prepared according to the method in Example 1 above.

[0081] Control group 3: 0.5g capsaicin was replaced with 0.5g azone according to the prescription of the active ingredient in Example 1 above, and then a gel patch was prepared according to the method in Example 1 above.

[0082] 1.3. Animal husbandry and experimental conditions

[0083] Animal husbandry, observation of pain behavior, establishment of an acute low back pain model of intervertebral disc degeneration (IDD), and application of topical medication were all conducted in the SPF-grade laboratory of the Experimental Animal Center of Southern Medical University. All rats were fed Co-60 radiation-sterilized feed and purified water. The normal laboratory temperature was 21–24℃, stable at 22℃; the relative humidity was 40%–70%, stable at 50%; the noise level was 37–45 dB; the gradient pressure difference was 20.0–31.0 Pa; the air exchange rate and air cleanliness met the standards; and the lighting was a 12-hour light / 12-hour dark cycle.

[0084] 2. Modeling Method

[0085] 2.1 Grouping

[0086] The rats were randomly divided into 7 groups according to the random number table method, 6 rats in each group. Including the blank group, the model group, the positive control group (2x4 cm 2 Wan Tong muscle and bone paste), the experimental group (2x4 cm 2 gel patch of Example 1), control group 1 (2x4 cm 2 Xinjia drug gel patch), control group 2 (2x4 cm 2 Huoluo Xiaolingdan gel patch), control group 3 (2x4 cm 2 nitrogen ketone gel patch). The biological rhythm was maintained in the feeding room and the behavior laboratory. The rats were placed in the experimental observation box for 30 minutes every day for 7 days before the experiment to adapt to the change of environment. The rats were changed every morning at 9 o'clock every day for 7 days after the experiment, and the condition of the rats was observed every day.

[0087] 2.2 Establishment of rat model of lower back pain caused by intervertebral disc degeneration

[0088] The lumbar intervertebral disc degeneration model was prepared by full-thickness annulus fibrosus puncture method (see Figure 1 ). The SD rats were anesthetized with 2% isoflurane mixed with oxygen gas. After successful anesthesia, the rat was fixed in supine position, and the specific position of the intervertebral disc between L4-L5 was determined by palpation and radiology. The skin was prepared with a 3cmx5cm incision centered on the midline of the abdomen, cleaned, and iodophor was used for disinfection. The left lateral abdominal external side of L4-5 lumbar intervertebral disc was exposed by left transabdominal spinal approach. Care should be taken not to damage the main blood vessels near the lumbar vertebrae. The L4-5 intervertebral space was exposed, and a 21G puncture needle was selected. The needle was inserted from the lateral side of the annulus fibrosus, parallel to the cartilage endplate, at an angle of about 45° to the sagittal plane of the spine. A self-made needle cap was used as a protective device to control the depth of needle insertion, about 3mm. Full-thickness needle puncture was performed, and the puncture needle was removed after staying for 10s. The peritoneum and abdominal wall were sutured with 3-0 silk suture, and the skin was sutured with 4-0 nylon suture, and attention should be paid to layer-by-layer suturing. The rats were closely monitored to ensure that there were no intraoperative complications. The rats in the blank group were only exposed to the intervertebral disc operation area, and were not modeled, and did not need to be punctured. For IDD rats treated with the positive control group, the experimental group, control group 1, control group 2 and control group 3, the corresponding patches were pasted on the L4-5 intervertebral disc of the rat waist for 14 consecutive days. For the model group rats, blank patches without drugs were pasted. After operation, 800,000 U / (kg·d) of penicillin was maintained by intramuscular injection for 3 consecutive days to prevent wound infection. In order to explore the best administration time, we explored the intervertebral disc degeneration of rats two weeks after modeling and four weeks after modeling in the early stage, and according to the results we decided to intervene with the patch two weeks after modeling. After the pain behavior and MRI imaging measurement at the 4-week time point after modeling, the abdominal aortic blood of the rats was collected, and the lumbar vertebrae was separated for morphological and serological analysis.

[0089] 3, Index detection

[0090] 3.1 MRI imaging changes

[0091] Each experimental group of rats with 2% isoflurane and oxygen mixture gas anesthesia. Using PharmaScan 70 / 16US (Brook Biospin MRI Gmbh) on the double-tuned volume RF coil scanning rats L4-5 region. Sagittal T2 weighted image using the following method to collect parameters: using spin echo sequence to obtain sagittal T2 weighted image covering the entire disc region, parameters as follows: fat saturation; repetition time / effective echo time, 2500 / 27 ms; field of view, 50 x 50 mm; matrix, 320 x 320; slice thickness, 0.5 mm; interval, 0 mm; slice number, 16; total scan number, 1 (total scan time, 16.24 min). MRI immediately using 7-T imager (Bruker BioSpec System, Bruker Corporation, Billerica, MA, USA) and BGA-S gradient coil scanning rats L4 / 5 lumbar spine. Get a representative MRI image of the blank group (A), model group (B), experimental group (C), positive drug group (D), control group 1 (E), control group 2 (F), control group 3 (G) rats, experimental results (see Figure 2 )

[0092] 3.2 Behavioral pain threshold indicators

[0093] Respectively, one day before modeling and two weeks after modeling and two weeks after administration, a total of 3 time points were detected. The hot plate pain test instrument was set to a constant temperature of 55℃, and the rats were placed in a translucent organic glass box (25x25x40cm) of the hot plate pain test instrument. The bottom was black anodized aluminum plate, the timing button was opened, and the rat appeared to be positive reaction, and the time from the start to the positive reaction was recorded. The cutoff time was 30 seconds to avoid heat injury to the rats. Each rat was measured 5 times at the same time point, with an interval of 15 minutes between each measurement. The highest value and the lowest value were removed, and the average value was taken. The thermal pain threshold of each group of rats was detected at 8:00-12:30 am.

[0094] 3.3 Hemorheology indicators

[0095] After 2 weeks of application, the rats in each group were anesthetized, and 10 ml of blood was taken from the abdominal aorta, of which 5 ml was injected into a heparin tube and shaken well. The heparin tube was placed in ice and sent to the First Affiliated Hospital of Guangzhou University of Chinese Medicine for hemorheology experiment. Cooperate with the First Affiliated Hospital of Guangzhou University of Chinese Medicine to determine: whole blood viscosity high shear (200 / s), medium shear (30 / s), low shear (3 / s), whole blood viscosity and plasma viscosity indicators.

[0096] 3.4 Effect on serum inflammatory factors TNF-α, IL-1β

[0097] The remaining abdominal aorta blood 5 mL was collected, centrifuged at 3500 r / min for 15 min, and the supernatant was absorbed and divided into 100 μl per tube as a specimen, which was stored at -80℃. The inflammatory levels of TNF-α and IL-1β in rat serum were measured by ELISA according to the manufacturer's instructions.

[0098] 3.5 Pathology HE staining index

[0099] Histology is mainly evaluated and analyzed by HE staining of lumbar intervertebral disc tissue, which is the most accurate method for evaluating lumbar intervertebral disc degeneration, and is also the gold standard in clinical pathological diagnosis. The steps of HE staining are as follows: 42 rats were sacrificed 4 weeks after operation, and the L4-5 intervertebral discs of rats were dissected. Fix in 4% formaldehyde buffer for 48 hours, and decalcify in 10% EDTA (pH 7.4) for 4 weeks. Next, embed the decalcified tissue in paraffin and cut into pathological sections with a thickness of 5 μm. Use HE staining sagittal section to evaluate the morphology of nucleus pulposus and annulus fibrosus in intervertebral disc, and use optical microscope for evaluation.

[0100] 3.6 Statistical examination

[0101] All data were statistically analyzed by SPSS21.0 statistical software, and the data were expressed as "mean ± standard deviation". The differences between groups were compared by one-way ANOVA, and the multiple comparisons between groups were compared by LSD method (when variance is equal) or Dunnett's method (when variance is not equal). P<0.05 indicates significant difference, and P<0.01 indicates extremely significant difference.

[0102] 4、Experimental results:

[0103] 4.1 Increase the intervertebral disc degeneration of rats L4-5 intervertebral disc nucleus pulposus area and signal intensity

[0104] See Figure 2(A) In the blank control group, the L4-5 intervertebral disc intervertebral space height was normal, the nucleus pulposus area was full and oval, and the signal intensity was high. (B) In the model group, the L4-5 intervertebral disc intervertebral space height disappeared, the nucleus pulposus area disappeared, and the MRI signal intensity was low. The intervertebral disc showed pathological features of degeneration and stenosis. (C) In the experimental group, the L4-5 intervertebral disc showed medium signal intensity, the nucleus pulposus area was partially reduced, but the central transparent cavity was still present, and the intervertebral space was slightly narrowed. The intervertebral space height and signal intensity were higher than those in the model group. (D) In ​​the positive control group, the L4-5 intervertebral disc nucleus pulposus area and signal intensity were higher than those in the model group but lower than those in the experimental group. (E) In the control group 1, the L4-5 intervertebral disc nucleus pulposus area and signal intensity were higher than those in the model group but lower than those in the experimental group. (F) In the control group 2, the L4-5 intervertebral disc nucleus pulposus area and signal intensity were higher than those in the model group but lower than those in the experimental group. (G) In the control group (3), the nucleus pulposus area and signal intensity of the L4-5 intervertebral disc were higher than those in the model group but lower than those in the experimental group. These experimental results suggest that the patented gel patch can significantly delay the pathological progression of intervertebral disc degeneration in rats.

[0105] 4.2 Improving the behavioral pain threshold in rats with intervertebral disc degeneration

[0106] The pain threshold results of rats with intervertebral disc degeneration in each group were recorded during the study period, and the results are shown in Table 1. Compared with the blank group, the screeching time of rats in the model group, experimental group, positive control group, control group 1, control group 2, and control group 3 was significantly shortened three weeks after modeling, indicating that the pain threshold of rats was significantly reduced after modeling. * P < 0.05, consistent with the pain sensitivity characteristics of patients with low back pain caused by intervertebral disc degeneration. Two weeks after administration, compared with the model group, the positive control group, control group 1, control group 2, and control group 3 showed significant efficacy (P < 0.05). ★ P<0.05), the experimental group showed extremely significant efficacy ( ★★ P<0.01); Compared with the experimental group, the experimental group significantly prolonged the duration of rat screech in control groups 1, 2, 3 and the positive control group. ▲ (P < 0.05) increased the pain threshold in rats; the above experimental results suggest that the patented gel patch can prevent pain response caused by intervertebral disc degeneration and improve the pain tolerance of rats.

[0107] Table 1. Results of behavioral pain threshold experiments in rats with intervertebral disc degeneration in each group.

[0108]

[0109] Note 1: Compared with the blank group, * p<0.05.2, compared with the experimental group, ▲p<0.05.3, compared with the model group, ★ p<0.05, ★★ p<0.01

[0110] 4.3 Results of hemorheological experiments on rats with improved intervertebral disc degeneration

[0111] The results of blood rheology tests on rats with intervertebral disc degeneration during the study period are shown in Table 2. Compared with the control group, the model group showed significantly increased high, medium, and low shear whole blood viscosity and plasma viscosity. * P<0.05, indicating increased blood viscosity after modeling, consistent with the signs of qi stagnation and blood stasis in patients with low back pain caused by lumbar disc degeneration. Two weeks after administration, compared with the model group, the positive control group, control group 1, control group 2, and control group 3 showed significant efficacy. ★ P<0.05), the experimental group showed extremely significant efficacy ( ★★ P<0.01); Compared with the experimental group, the experimental group significantly reduced the hemorheological parameters of rats in control groups 1, 2, 3, and the positive control group. ▲ P < 0.05) enhances the blood circulation and blood stasis removal ability of rats; the above experimental results suggest that the patented gel patch can significantly improve the local blood microcirculation of rats with intervertebral disc degeneration and prevent the occurrence of blood stasis reaction.

[0112] Table 2. Hemorheological results of rats with intervertebral disc degeneration in each group.

[0113] Group High shear (mPa.s) Medium shear (mPa.s) Low shear (mPa.s) Plasma (mPa.s) Model group 6.69±0.62* 10.06±0.70* 45.65±5.75* 5.84±0.37* Blank group 4.86±0.18 6.68±0.39 34.02±3.02 3.85±0.39 Experimental group 4.99 ± 0.19 ★★ ]] 6.92 ± 0.26 ★★ ]] 35.13 ± 1.13 ★★ ]] 3.98 ± 0.16 ★★ ]] Positive control group 5.97 ± 0.10 ★▲ ]] 8.35 ± 0.43 ★▲ ]] 43.76 ± 1.13 ★▲ ]] 4.94 ± 0.36 ★▲ ]] Control group 1 5.95 ± 0.96 ★▲ ]] 8.70 ± 1.28 ★▲ ]] 42.12 ± 1.86 ★▲ ]] 4.89 ± 0.85 ★▲ ]] Control group 2 5.60 ± 1.29 ★▲ ]] 8.69 ± 0.93 ★▲ ]] 42.03 ± 2.52 ★▲ ]] 4.92 ± 0.57 ★▲ ]] Control group 3 5.75 ± 1.13 ★▲ ]] 7.98 ± 0.93 ★▲ ]] 42.53 ± 1.72 ★▲ ]] 4.86 ± 0.76 ★▲ ]]

[0114] Note 1: Compared with the blank group, * p<0.05.2, compared with the experimental group, ▲ p<0.05.3, compared with the model group, ★ p<0.05, ★★ p<0.01

[0115] 4.4 Reduce serum levels of inflammatory factors TNF-α and IL-1β in rats with intervertebral disc degeneration

[0116] IL-1β and TNF-α are classic inflammatory mediators in the cytokine network, leading to increased sensitivity to local pain and exacerbating low back pain. The results of serum inflammatory factor detection in rats with intervertebral disc degeneration during the study period are shown in Table 3. The serum levels of IL-1β and TNF-α in the model group were significantly higher than those in the control group (*P<0.05), indicating that after modeling, the content of inflammatory factors in the rat serum increased, resulting in an inflammatory pain response. Two weeks after drug administration, compared with the model group, the positive control group, control group 1, control group 2, and control group 3 showed significant efficacy in reducing serum inflammatory factor levels.★ P<0.05), the experimental group suggests a very significant drug effect ★★ P<0.01); wherein, the control group 1, the control group 2, the control group 3, the positive control group and the experimental group comparison suggests that the experimental group can significantly reduce the serum IL-1β and TNF-α levels of intervertebral disc degeneration rats( ▲ P<0.05), enhance the anti-inflammatory ability of rats; the above experimental results suggest that the patent gel patch can significantly inhibit the overexpression of pain-related inflammatory factors in the serum of intervertebral disc degeneration rats.

[0117] Table 3 Serum inflammatory factor results of intervertebral disc degeneration rats in each group

[0118]

[0119]

[0120] Note: 1. Compared with the blank group, * p<0.05.2. Compared with the experimental group, ▲ p<0.05.3. Compared with the model group, ★ p<0.05, ★★ p<0.01

[0121] 4.5 Delaying the pathological progression of intervertebral disc degeneration in rats

[0122] The results of hematoxylin-eosin staining of each group of intervertebral disc degeneration rats during the study are shown in Figure 3 : The blank group of rats has normal intervertebral disc structure, and the nucleus pulposus tissue occupies about 75% of the total intervertebral disc area. The nucleus pulposus cells are oval and vacuolated, and are evenly distributed. The boundary between the elliptical nucleus pulposus tissue and the annulus fibrosus is clear. The intervertebral disc structure of the model group of rats shows obvious degenerative changes. The elliptical structure of the nucleus pulposus tissue disappears, the annulus fibrosus is arranged in disorder, the boundary between the annulus fibrosus and the nucleus pulposus tissue is completely interrupted, and the structure is not clear. The normal nucleus pulposus tissue of the experimental group of rats occupies about 45% of the intervertebral disc area, the edge is relatively smooth and uniform, and the nucleus pulposus part shrinks, but the central transparent cavity still exists. The annulus fibrosus is arranged relatively in order, and the boundary between the annulus fibrosus and the nucleus pulposus tissue is partially clear. The nucleus pulposus tissue of the positive drug group, the control group 1, the control group 2, and the control group 3 disappears, and occupies about 25% of the intervertebral disc area, which is gradually replaced by fibrocartilage tissue. The above experimental results suggest that the patent gel patch can significantly delay the progression of intervertebral disc degeneration in rats.

[0123] II. 3 2 Analysis of variance of factorial design to study synergistic effect

[0124] (I) Experimental method

[0125] The experiment respectively takes the classic famous prescription Huoluo Xialingdan (Angelica sinensis, Salvia miltiorrhiza, Frankincense, Myrrh) and the newly added drugs (Eucommia ulmoides, Dipsacus asper, Loranthus parasiticus, Curcuma longa) as the analysis factors, each factor respectively takes the respective dosage of Huoluo Xialingdan and the newly added drugs in the examples 1, 2 and 3 in the examples as a level to carry out two-factor three-level factorial design, and then the experimental data is subjected to variance analysis. The experimental drugs of the 9 experimental groups are all gel patches, and the specific preparation method refers to the example 1 in the above example 1; the specific research method refers to the above “I. Pharmacodynamic test”.

[0126] All data are statistically analyzed by SPSS21.0 statistical software, and the data are represented as “mean ± standard deviation ”; the main effects of Huoluo Xialingdan and the newly added drugs and the interaction effects of the combination of the two are analyzed by variance analysis method of 3 2 factorial design; P<0.05 indicates significant difference, and P<0.01 indicates extremely significant difference.

[0127] (II) Experimental results

[0128] (1) Behavioral score of intervertebral disc degeneration rats

[0129] Table 4 Effect of combination of Huoluo Xialingdan and newly added drugs on behavioral pain threshold

[0130]

[0131]

[0132] The main effects and interaction effects of each factor of the data in Table 4 are statistically analyzed, and the results are shown in Table 5. The P values of the two main effects are both less than 0.01, and the P value of the interaction effect is less than 0.05, which all have statistical significance, indicating that there is an interaction between Huoluo Xialingdan and the newly added drugs, and the combination has an inhibitory effect on the pain behavior of rats with lower back pain caused by intervertebral disc degeneration.

[0133] Table 5 Variance analysis table of behavioral pain threshold score of combination of Huoluo Xialingdan and newly added drugs

[0134]

[0135] (2) Hemorheology index of intervertebral disc degeneration rats

[0136] ① Observation index: high shear

[0137] Table 6 Effect of combination of Huoluo Xialingdan and newly added drugs on high shear of hemorheology

[0138]

[0139] The main effect and interaction effect of each factor of the data in Table 6 were statistically analyzed, and the results are shown in Table 7. The P values of the two main effects are less than 0.01, and the P value of the interaction effect is less than 0.05, which are statistically significant. It is indicated that the Huoluo Xiaolingdan and the new added drugs have interaction, can reduce the high shear whole blood viscosity of the lower back pain rats caused by intervertebral disc degeneration, and have the effect of promoting blood circulation to remove blood stasis.

[0140] Table 7 Variance analysis table of the effect of the combination of Huoluo Xiaolingdan and the new added drugs on the high shear of hemorheology

[0141]

[0142] ②Observation index: medium shear

[0143] Table 8 Effect of the combination of Huoluo Xiaolingdan and the new added drugs on the medium shear of hemorheology

[0144]

[0145] The main effect and interaction effect of each factor of the data in Table 8 were statistically analyzed, and the results are shown in Table 9. The P values of the two main effects are less than 0.01, and the P value of the interaction effect is less than 0.05, which are statistically significant. It is indicated that the Huoluo Xiaolingdan and the new added drugs have interaction, can reduce the medium shear whole blood viscosity of the lower back pain rats caused by intervertebral disc degeneration, and enhance the ability of promoting blood circulation to remove blood stasis.

[0146] Table 9 Variance analysis table of the effect of the combination of Huoluo Xiaolingdan and the new added drugs on the medium shear of hemorheology

[0147]

[0148] ①Observation index: low shear

[0149] Table 10 Effect of the combination of Huoluo Xiaolingdan and the new added drugs on the low shear of hemorheology

[0150]

[0151]

[0152] The main effect and interaction effect of each factor of the data in Table 10 were statistically analyzed, and the results are shown in Table 11. The P values of the two main effects are less than 0.01, and the P value of the interaction effect is less than 0.05, which are statistically significant. It is indicated that the Huoluo Xiaolingdan and the new added drugs have interaction, can reduce the low shear whole blood viscosity of the lower back pain rats caused by intervertebral disc degeneration, and enhance the ability of promoting blood circulation to remove blood stasis.

[0153] Table 11 Variance analysis table of the effect of the combination of Huoluo Xiaolingdan and the new added drugs on the low shear of hemorheology

[0154]

[0155] (3) Serum inflammatory factor indexes of intervertebral disc degeneration rats

[0156] ① Observation index: TNF-α content

[0157] Table 12 Effect of Huoluo Xiaolingdan combined with new drugs on serum inflammatory factor TNF-α

[0158]

[0159] The statistical analysis of main effect and interaction effect of each factor on the data in Table 12 is shown in Table 13, the P value of two main effects is less than 0.01, the P value of interaction effect is less than 0.05, the differences have statistical significance, which shows that there is interaction between Huoluo Xiaolingdan and new drugs, and the drugs have therapeutic effect on the decrease of serum inflammatory factor TNF-α content of intervertebral disc degeneration rats and enhance the anti-inflammatory ability.

[0160] Table 13 Variance analysis table of effect of Huoluo Xiaolingdan combined with new drugs on serum inflammatory factor TNF-α

[0161]

[0162] ② Observation index: IL-1β content

[0163] Table 14 Effect of Huoluo Xiaolingdan combined with new drugs on serum inflammatory factor IL-1β

[0164]

[0165] The statistical analysis of main effect and interaction effect of each factor on the data in Table 14 is shown in Table 15, the P value of two main effects is less than 0.01, the P value of interaction effect is less than 0.05, the differences have statistical significance, which shows that there is interaction between Huoluo Xiaolingdan and new drugs, and the drugs have therapeutic effect on the decrease of serum inflammatory factor IL-1β content of intervertebral disc degeneration rats and enhance the anti-inflammatory ability.

[0166] Table 15 Variance analysis table of effect of Huoluo Xiaolingdan combined with new drugs on serum inflammatory factor IL-1β

[0167]

[0168] Three, experimental conclusion

[0169] In summary, the combination of Huolüoxì Lingdan and Xinka has synergistic effect in the treatment of lower back pain caused by intervertebral disc degeneration, and the effect is significantly better than that of the positive drug and single Huolüoxì Lingdan or Xinka. It can greatly relieve the symptoms of lower back pain caused by intervertebral disc degeneration and delay the pathological progression of intervertebral disc degeneration.

Claims

1. A gel patch for treating lower back pain caused by intervertebral disc degeneration, the gel patch comprising a backing layer, a drug reservoir layer, and an anti-adhesion layer, wherein the drug reservoir layer comprises an active ingredient, a matrix, and a penetration enhancer; characterized in that, The effective components are volatile oils and fluid extracts made from the following raw materials in parts by weight: turmeric 17.8–21.7%, angelica 11.8–14.5%, frankincense 11.8–14.5%, myrrh 11.8–14.5%, salvia miltiorrhiza 11.8–14.5%, dipsacus asper 8.3–10.1%, mulberry mistletoe 8.3–10.1%, and eucommia ulmoides 8.3–10.1%; The active ingredient is prepared by the following method: Take the raw material and extract it twice by steam distillation. For the first extraction, use 8 times the amount of water and steam distill for 2 hours, collecting the volatile oil and the liquid separately. Add 6 times the amount of water to the residue and steam distill for 1 hour, collecting the volatile oil and the liquid separately. Combine the liquids from the two steam distillations and evaporate them using a rotary evaporator to obtain a fluid extract with a concentration equivalent to 2 g / mL of the raw drug. The obtained fluid extract and the volatile oil extracted by the two steam distillations are then ready.

2. The gel patch for treating lower back pain caused by intervertebral disc degeneration according to claim 1, characterized in that, The active ingredients are volatile oils and fluid extracts made from the following raw materials in parts by weight: turmeric 19.6%, angelica 13.2%, frankincense 13.2%, myrrh 13.2%, salvia miltiorrhiza 13.2%, dipsacus 9.2%, mulberry mistletoe 9.2%, and eucommia ulmoides 9.2%.

3. The gel patch for treating lower back pain caused by intervertebral disc degeneration according to claim 2, characterized in that, The penetration enhancer is capsaicin.

4. The gel patch for treating lower back pain caused by intervertebral disc degeneration according to claim 3, characterized in that, The ointment of the drug storage layer is prepared by the following method: (1) Take 0.15% of the weight of the fluid extract, add 150 times the amount of water and swell in a 90°C water bath. Then add the fluid extract and stir evenly. After that, add 25% of the weight of the fluid extract and seal and let it stand to swell to obtain phase A. (2) Dissolve 10.63% by weight of citric acid and 5.31% by weight of aluminum glycinate in water, then dissolve capsaicin in ethanol, then combine the two and add the volatile oil and 75% by weight of glycerol, stir evenly to obtain phase B; (3) In a 50°C water bath, the A phase prepared in step 1 and the B phase prepared in step 2 are mixed evenly to obtain a mixed solution; (4) Add sodium polyacrylate to the mixed solution prepared in step 3 at 14.86% of the weight of the active ingredient, and continue to mix it evenly in a water bath at 50°C to obtain the ointment of the drug storage layer.

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