Application of zanthoxylum bungeanum maxim extract in preparation of medicine for treating skin diseases

Abstract

The invention relates to the field of modern application of traditional Chinese medicines, in particular to application of a pepper extract in preparation of a medicine for treating skin diseases. Furthermore, the medicine can also be used for treating itching and pain caused by skin diseases such as herpes zoster pain, atopic dermatitis, eczema, neurodermatitis, senile skin itching, psoriasis and diabetic foot. Wherein the postherpetic pain comprises pain, pruritus and / or pain, tactile hypersensitivity and the like. According to the invention, the zanthoxylum bungeanum extract obtained by preferably using a supercritical extraction method is zanthoxylum oleylamine containing active ingredients such as zanthoxylum amide and zanthoxylum bungeanum volatile oil, and the effect of the zanthoxylum bungeanum extract is obviously superior to that of monomers such as hydroxyl-alpha-sanshool; the Chinese prickly ash extract is a pure natural Chinese prickly ash extract component, is easy to accept by patients, has a high compliance rate on medical advice, and can effectively solve the problem of lack of special medicines for treating postherpetic pain and the like in the prior art, and the Chinese prickly ash extract is a pure natural Chinese prickly ash extract component which is a pure natural Chinese prickly ash extract component which is easy to accept by the patients and has a high compliance rate on medical advice.

Description

Technical Field

[0001] The invention relates to the field of biomedicine, and in particular to the application of Zanthoxylum bungeanum extract in the development of new drugs, specifically to the application in the preparation of drugs for treating post-herpetic pain and the like.

[0002] This application claims the priority of Chinese invention patent application No. 2023108537517. The application date of the priority patent is 20230712. The name of the invention of the priority application is "Application of a Zanthoxylum bungeanum extract in the preparation of a medicament for treating postherpetic pain". The entire contents of the prior application documents are incorporated into this application. Background Art

[0003] Postherpetic neuralgia (PHN) is the most common sequela of herpes zoster. It is generally believed that after the rash of herpes zoster subsides, the local skin is still painful and uncomfortable for more than one month, which is called postherpetic pain or postherpetic neuralgia, which manifests as paroxysmal or persistent burning pain, tingling, throbbing pain, and knife-cutting pain. It is more common in the elderly. Modern medicine believes that postherpetic neuralgia is related to the reduction of human immunity, and its incidence rate increases with age. It usually occurs after the herpes disappears, and postherpetic neuralgia appears in the primary skin area. It is reported that PHN can last for three months or even longer, seriously affecting the patient's quality of life and also bringing great burden to the patient's family and society.

[0004] In addition to pain (burning, electric shock-like pain), post-herpetic pain also includes skin tactile and / or sensory hypersensitivity, causing paresthesia, pain induced by light touch, ant-like sensation, itching, local tightening and numbness, etc. The specific manifestation is that the skin in the painful area cannot tolerate light touch, and even light touch of one's own clothes can cause extremely uncomfortable feeling. Studies have shown that herpes zoster virus infection can cause the nerve sheath to rupture, the virus is killed and basically disappears, and after the treatment of damage such as rash is restored, the damaged nerve cells are prone to continuous spontaneous discharge due to excessive excitement, or the sensitivity of nerve cells increases, making it impossible to treat post-herpetic pain through antiviral drugs. It is generally believed that the pain and sensory hypersensitivity of post-herpetic pain have a very complex mechanism, which may be related to the loss of nerve fiber damage caused by long-term viral chronic infection, increased activity of sensory afferent fibers, abnormal discharges and the resulting central hyperexcitement.

[0005] The mechanism and manifestation of pain caused by herpes zoster itself and postherpetic pain are different. The pain caused by herpes zoster itself is caused by factors such as nerve inflammation during the herpes period, and usually only manifests as spontaneous burning pain; while the main cause of postherpetic pain is the abnormal activity of free nerve endings under the skin, damage to peripheral nerve plexuses and central dorsal horn nerve cells, etc., which usually transforms into persistent spontaneous pain or intermittent spontaneous pain, allodynia, etc.

[0006] Most of the existing drugs for treating PHN target pain and use analgesics for intervention. Some clinicians use drugs such as pregabalin, lidocaine, and capsaicin for treatment, which block nerve signal transmission through local anesthesia to reduce sensitivity and eliminate residual pain. However, the efficacy is not significant enough, and long-term use of such drugs has certain toxic side effects, which slows down the recovery and improvement of the patient's actual health status.

[0007] Itch is the most common symptom of skin diseases, which not only affects the body (such as skin damage, secondary infection, scarring and reduced sleep, etc.), but can also lead to emotional disorders such as anxiety and depression. The incidence of chronic pruritus is extremely high, even reaching 60% in the elderly, affecting a very wide range of people. Dermatogenic pruritus accounts for the largest proportion of chronic pruritus, and is common in atopic dermatitis, eczema, psoriasis, neurodermatitis, eczema, psoriasis, sequelae of herpes zoster and chronic urticaria.

[0008] Atopic dermatitis is a chronic, recurrent, inflammatory skin disease, with the main clinical manifestations of eczematous skin lesions, severe itching, repeated attacks and a long course of disease. Its pathogenesis is complex and is closely related to factors such as skin barrier dysfunction, immune abnormalities, skin flora disorders, and family inheritance. According to surveys, the global prevalence of the disease in children is 15% to 30%, and in adults it is 7% to 10%. At least 230 million people are suffering from the disease worldwide, and it is increasing year by year. There is currently no radical treatment for atopic dermatitis. Local medication or combined systemic medication can be selected according to the severity of the disease. According to the guidelines, topical glucocorticoids are the first-line treatment for atopic dermatitis, and calcineurin inhibitors are important anti-inflammatory drugs; the Chinese guidelines believe that external preparations such as zinc oxide oil (paste) and black bean distillate ointment also have a certain antipruritic effect. Systemic medications mainly include antihistamines, immunosuppressants, glucocorticoids, biological agents, JAK inhibitors, etc. Traditional Chinese medicine believes that atopic dermatitis is caused by a combination of internal and external evils. The internal causes are spleen deficiency and stomach heat, spleen and lung qi deficiency, and hyperactivity of heart fire. The external causes are wind, dampness, and heat that fight against each other in the skin, which turn into heat over time. The dampness and heat accumulate in the skin pores and cause the disease. The treatment principles are to strengthen the spleen and eliminate dampness, clear away heat and cool blood, promote blood circulation and remove blood stasis, and dispel wind and relieve itching.

[0009] Neurodermatitis is a common chronic neurological dysfunction disease of the skin, also known as chronic simple lichen, with long course of disease, repeated skin itching, skin lichenification and other main characteristics. In the population, the incidence of neurodermatitis is about 12%, and the number of female patients is more than that of male patients. It is mostly limited to the back of the neck, elbows, popliteal fossa, and sacral coccyx. It can also be multi-site or systemic. Because the disease is difficult to cure and easy to relapse, patients are prone to insomnia, irritability, anxiety and other symptoms, which greatly affect the quality of life of patients. Modern medicine believes that neurodermatitis is mostly caused by local skin immune pathological reactions caused by internal and external stimulation such as neuropsychiatric psychology, immunity, and endocrine. Traditional Chinese medicine believes that neurodermatitis is mostly caused by rheumatic heat blocking the skin at the beginning. When encountering emotional frustration, heart fire inflammation, spleen and stomach dampness and heat, the qi and blood circulation is dysfunctional, the skin is stagnant, the disease is long-term, the yin of the ying is damaged, and the blood deficiency causes wind and dryness, and the skin is malnourished. There is currently a lack of safe and effective specific treatment drugs for neurodermatitis. Most clinical treatments use topical glucocorticoids such as halometasone cream and flumethasone ointment or combined with antihistamines. However, due to the long course of the disease, topical glucocorticoid preparations are prone to cause adverse reactions such as local skin atrophy and capillary dilation, and safer and more effective treatment drugs are urgently needed.

[0010] Zanthoxylum bungeanum, also known as Sichuan pepper, Sichuan pepper or Qin pepper, is a plant of the Rutaceae family. Zanthoxylum bungeanum is a traditional Chinese spice and Chinese medicinal material. In 2002, it was confirmed by the Ministry of Health as a dual-purpose food and medicine. In terms of food, Zanthoxylum bungeanum has always been loved by people for its unique strong aroma and lingering numbing taste, and has become a must-have seasoning in the kitchen; in terms of medicine, Zanthoxylum bungeanum is spicy and warm in nature, enters the stomach, kidney, and spleen meridians, and has the effects of killing insects, relieving itching, warming the middle and relieving pain, and dispelling cold. Although Zanthoxylum bungeanum contains a variety of active substances such as volatile oils, amides, and flavonoids, and has high edible and medicinal value, my country has not yet formed a complete Zanthoxylum bungeanum industry chain. It is mainly based on primary products such as Zanthoxylum bungeanum granules, Zanthoxylum bungeanum oil, and Zanthoxylum bungeanum powder. The degree of fine processing is very low, and there is little development and utilization of products specifically for Zanthoxylum bungeanum active substances. This is actually a waste of existing resources.

[0011] Zanthoxylum bungeanum volatile oil is an oily liquid component that can evaporate with steam distillation and is the main chemical component of Zanthoxylum bungeanum. The volatile oil content in Zanthoxylum bungeanum is about 0.7% to 9.0%, and the main chemical components are small molecules such as alcohols, olefins, ketones, terpenes, esters, epoxides and aldehydes. Zanthoxylum bungeanum fruits, roots, leaves, flowers and other parts contain volatile oils, among which the fruit has the highest content. Zanthoxylum bungeanum volatile oil is a transparent liquid with a color of light yellow to orange-yellow. It is a mixture with complex components and an aromatic smell. It is volatile at room temperature, insoluble in water but easily soluble in various organic solvents, such as ether and ethanol. Studies have found that Zanthoxylum bungeanum volatile oil has anti-tumor, antioxidant, insecticidal, antibacterial and antiseptic, anti-inflammatory and analgesic effects, and inhibits smooth muscle contraction.

[0012] Zanthoxylum bungeanum is rich in alkaloids, among which the amide alkaloid components are the flavor and active substance basis of Zanthoxylum bungeanum, and are also the characteristic components of Zanthoxylum bungeanum. The content of sanshool components is rich, including α-sanshool, hydroxy-α-sanshool, β-sanshool, hydroxy-β-sanshool, γ-sanshool, hydroxy-γ-sanshool, etc., which can be extracted by solvents such as ethanol, methanol, n-butanol, ethyl acetate, etc. Although Zanthoxylum bungeanum has a long history of consumption, it is not considered to be an effective drug for the treatment of PHN. In addition, although there have been studies in the prior art on the use of some components of Zanthoxylum bungeanum for anti-inflammatory and analgesic purposes, since the pain of PHN is not directly related to inflammation, common anti-inflammatory analgesics such as acetaminophen and ibuprofen have no clear therapeutic effect on PHN. The pain and hypersensitivity of PHN have very complex mechanisms. Studies have found that it may be related to the loss of nerve fiber damage caused by long-term viral chronic infection, increased activity of sensory afferent fibers, abnormal discharges and the resulting central overexcitation. Summary of the invention

[0013] The purpose of the present invention is to overcome the problem of lack of effective therapeutic drugs for postherpetic pain in the prior art, and to provide an application of a Zanthoxylum bungeanum extract in the preparation of a drug for treating postherpetic pain.

[0014] In addition, the drug can also treat itching and pain caused by herpes zoster pain, herpes zoster pain, atopic dermatitis, eczema, neurodermatitis, senile skin itching, psoriasis and diabetic foot.

[0015] The present invention relates to application of Zanthoxylum bungeanum extract in preparing medicine.

[0016] Specifically, it is an application of Zanthoxylum bungeanum extract in the preparation of medicines for treating skin diseases.

[0017] Furthermore, the present invention also aims to provide a use of a Zanthoxylum bungeanum extract in the preparation of a medicament for treating postherpetic pain, herpes zoster pain, atopic dermatitis, allergic dermatitis, neurodermatitis or senile skin pruritus.

[0018] Furthermore, the purpose of the present invention is to provide a Zanthoxylum bungeanum extract with preferred therapeutic effect for use in the preparation of a medicament for treating itching and pain caused by at least one of the skin diseases selected from the group consisting of postherpetic pain, herpes zoster pain, atopic dermatitis, allergic dermatitis, neurodermatitis or senile pruritus.

[0019] Preferably, the Zanthoxylum bungeanum extract has a better technical effect of treating itching and pain caused by skin diseases such as postherpetic pain, herpes zoster pain, atopic dermatitis, eczema, neurodermatitis, senile pruritus, psoriasis and diabetic foot compared to other Zanthoxylum bungeanum extracts or effective ingredients of Zanthoxylum bungeanum.

[0020] Preferably, the use of Zanthoxylum bungeanum extract in the preparation of a medicament is a medicament for treating itching and pain caused by at least one of eczema, psoriasis or diabetic foot.

[0021] Furthermore, the present invention also aims to provide a method for preparing the Zanthoxylum bungeanum extract having the preferred therapeutic effect.

[0022] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0023] The invention discloses an application of Zanthoxylum bungeanum extract in the preparation of medicines, wherein the medicines are medicines for treating itching and pain caused by skin diseases such as post-herpetic pain, herpes zoster pain, atopic dermatitis, eczema, neurodermatitis, senile pruritus, psoriasis and diabetic foot.

[0024] The pharmaceutical application of the pepper extract of the present invention has a significant therapeutic effect on the treatment of post-herpetic pain, etc., and realizes the innovation of pharmaceutical application of active ingredients of pure natural plant extracts, which well solves the problem of lack of targeted therapeutic drugs for post-herpetic pain. At the same time, skin diseases such as atopic dermatitis, eczema, neurodermatitis, senile skin pruritus, psoriasis and diabetic foot also have obvious symptoms such as itching, pain, skin hypersensitivity, etc., so it can be predicted that the pepper extract also has a good effect on itching and pain caused by skin diseases such as atopic dermatitis, eczema, neurodermatitis, senile skin pruritus, psoriasis and diabetic foot.

[0025] The analgesics and local anesthetics commonly used in the prior art for the treatment of PHN are not stable enough, and local anesthetics block the transmission of nerve sensations. Although patients no longer feel unbearable sequelae pain when taking the medicine, they also greatly reduce the skin's daily tactile perception ability, affecting the quality of life, and long-term use has obvious side effects, which is not good for the patient's health. Therefore, the present invention uses Zanthoxylum bungeanum extract to prepare a drug for the treatment of PHN, solves the problem of the lack of drugs for PHN in the prior art, and avoids the safety risk caused by the abuse of local anesthetics and analgesics in certain middle-aged and elderly patients.

[0026] Further, the post-herpetic pain includes one or more of pain, allodynia, and tactile hypersensitivity, and further, it may or may not be combined with pruritus. The use of Zanthoxylum bungeanum extract in the preparation of a drug for the treatment of PHN pain or tactile hypersensitivity, allodynia, and with or without pruritus can well exert a selective therapeutic effect on post-herpetic pain.

[0027] After the herpes zoster lesions heal, the skin often develops tactile / sensory hypersensitivity, which affects daily life. By using Zanthoxylum bungeanum extract in the preparation of drugs to relieve tactile / sensory hypersensitivity caused by herpes zoster, the daily quality of patients during recovery can be improved. Zanthoxylum bungeanum extract treats PHN, relieves tactile / sensory hypersensitivity, and PHN-related pruritus, avoids anesthetic drugs from completely blocking nerve perception, and better ensures the health status and quality of life of patients.

[0028] There are few studies on PHN in the prior art, and few studies on the pathogenesis of tactile / sensory hypersensitivity caused by PHN, and the specific invention mechanism is unclear. The inventor unexpectedly discovered in the study that the drug made from Zanthoxylum bungeanum extract can relieve tactile / sensory hypersensitivity in postherpetic pruritus. After scientific animal experimental research, it was verified that the efficacy is reliable and has the unique safety advantage of natural extract treatment. Experiments show that Zanthoxylum bungeanum extract can significantly increase the mechanical pain threshold of mice with neuroinflammation induced by allyl isothiocyanate (AITC).

[0029] The present invention relates to the use of a Zanthoxylum bungeanum extract in the preparation of a medicine for treating postherpetic pain. Further, the medicine can also treat pain and itching related to skin diseases such as herpetic pain, atopic dermatitis, eczema, psoriasis, allergic dermatitis, senile skin pruritus and neurodermatitis. Wherein, the postherpetic pain includes pain, itching and / or pain, tactile hypersensitivity and the like. The Zanthoxylum bungeanum extract preferably obtained by supercritical extraction is a Zanthoxylum bungeanum oil amine containing active ingredients such as Zanthoxylum bungeanum volatile oil. Its effect is significantly better than monomers such as Zanthoxylum bungeanum and hydroxy-α-sanshool, and is also significantly better than volatile oils extracted by steam distillation, or extracts extracted by water, or organic solvents such as ether and ethanol. It belongs to a pure natural Zanthoxylum bungeanum extract component, is easily accepted by patients, has a high compliance rate with medical advice, and can effectively solve the problem of lack of drugs specifically used for treating postherpetic pain in the prior art.

[0030] Furthermore, the Zanthoxylum bungeanum extract is prepared by supercritical extraction technology.

[0031] Further, the Zanthoxylum bungeanum extract is Zanthoxylum bungeanum, which is a Zanthoxylum bungeanum extract containing Zanthoxylum bungeanum amide components and Zanthoxylum bungeanum volatile oil extracted from Zanthoxylum bungeanum. The Zanthoxylum bungeanum solution of the present invention is better than the treatment of PHN with Zanthoxylum bungeanum amides or Zanthoxylum bungeanum volatile oil alone, which may be related to the amides and volatile oils in Zanthoxylum bungeanum amides cooperating with each other to promote penetration and act on subcutaneous nerve endings.

[0032] Further, the pepper oil amine is prepared by supercritical extraction technology. The pepper oil amine of the present invention is prepared by supercritical extraction technology. Compared with the volatile oil obtained by steam distillation and the pepper extract components obtained by water extraction, alcohol extraction, ether extraction, etc., and the amide chemical component monomers such as hydroxy-α-sanshool and hydroxy-β-sanshool, the pepper oil amine extracted by supercritical extraction has a more significant effect on the treatment of PHN, which has been verified by animal experiments. We believe that this may be due to the synergistic effect of the compatibility of the pepper amide component and the volatile oil in the pepper oil amine extracted by supercritical extraction. The water extraction method mainly separates the pepper polysaccharide component. Although the alcohol extraction method has a high extraction efficiency for volatile oil, the coordination relationship of the active ingredients in the obtained extract is not good, and the effect of treating PHN cannot be maximized. The ether extraction method is similar to the active ingredient composition obtained by the alcohol extraction method, and it also cannot play a good role in treating PHN. After comprehensive comparison, we chose the pepper oil amine prepared by supercritical extraction technology for the treatment of PHN.

[0033] Zanthoxylum bungeanum extract contains prickly ash amide, which can act on the Nav1.7 channel of neuron terminals, inhibit the influx of Na ions and block the transmission of pain, and can protect neurons and tactile receptors; combined with prickly ash volatile oil components, it further enhances the analgesic and transdermal penetration effects, thereby effectively relieving the symptoms of post-herpetic pain. After experimental analysis and research, prickly ash oil amine prepared by supercritical extraction technology contains prickly ash amide and prickly ash volatile oil components, and the two are in a suitable ratio, which can maximize the treatment of PHN. The application of prickly ash oleamine in drug preparation can achieve better quality assurance of drugs for the treatment of PHN, and the actual value of drug development is extremely great.

[0034] Furthermore, the postherpetic pain includes one or more of pain, allodynia, and tactile allergy, with or without itching.

[0035] The Zanthoxylum bungeanum extract is Zanthoxylum bungeanum oleylamine containing Zanthoxylum bungeanum amide components and Zanthoxylum bungeanum volatile oil.

[0036] Furthermore, the Zanthoxylum bungeanum is obtained by separating pure natural Zanthoxylum bungeanum through supercritical carbon dioxide extraction, and contains more than 50% of Zanthoxylum bungeanum amide components and more than 15% of Zanthoxylum bungeanum volatile oil components.

[0037] At the same time, the inventors also tried to compare zanthoxylum bungeanum with lidocaine (local anesthetic) and capsaicin (similar natural active ingredients). The results showed that zanthoxylum bungeanum is safer and less likely to cause adverse side effects.

[0038] Furthermore, the Zanthoxylum bungeanum extract of the present invention is obtained by pure natural extraction and separation, and belongs to pure natural Zanthoxylum bungeanum extract components. It does not use chemically synthesized small molecule drug components, is more easily accepted by patients, and the doctor's prescription can be more fully followed.

[0039] Furthermore, the supercritical extraction technology preparation refers to: adding the pepper raw material into an extraction tank and using carbon dioxide for supercritical extraction; during the extraction process, the CO2 temperature is controlled between 30 and 60°C.

[0040] Preferably, the CO2 temperature is controlled between 40 and 60°C.

[0041] Furthermore, the supercritical extraction technology preparation refers to: extracting pepper raw materials by supercritical carbon dioxide extraction, with an extraction temperature of 30-60°C, an extraction time of 2-4 hours, an extraction pressure of 15-30 MPa, and a carbon dioxide flow rate of 1.0-3m 3 / h.

[0042] Alternatively, the following parameters can be selected for the supercritical extraction technology preparation process: extraction temperature 40-60°C, extraction time 1-6 hours, extraction pressure 16-32MPa, and carbon dioxide flow rate 50-100kg / h.

[0043] Furthermore, the Zanthoxylum bungeanum is prepared by supercritical extraction technology, and the specific extraction process is as follows:

[0044] S1. Prepare the raw material of Sichuan pepper and fully dry it until the moisture content is less than 10-13%;

[0045] S2, crush the raw material of Sichuan pepper, add it into the extraction tank, and use carbon dioxide for supercritical extraction; during the extraction process, the temperature of CO2 is controlled between 30 and 60°C, and a two-stage evaporator is used to condense and collect the extract; after the carbon dioxide is extracted, it is recycled. Preferably, the temperature of CO2 is controlled between 30 and 60°C.

[0046] Furthermore, the medicine is an external preparation.

[0047] Preferably, the drug is any one of a tincture, a liniment, an ointment, a lotion, an emulsion, a film coating, a gel, a paste, a transdermal patch, a spray, and an aerosol.

[0048] Preferably, the drug is a liquid preparation and / or a semisolid preparation.

[0049] Furthermore, the drug contains pharmaceutically acceptable excipients.

[0050] Pharmaceutically acceptable excipients refer to inert substances that are administered together with active ingredients and facilitate the administration of active ingredients, including but not limited to any glidants, permeation enhancers, diluents, preservatives, dyes / colorants, flavor enhancers, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents or emulsifiers that are acceptable for human use and approved by the State Administration for Market Regulation.

[0051] Preferably, the auxiliary material comprises at least one or more of the following ingredients: gelatin, vegetable oil and polyethylene glycol.

[0052] Furthermore, the zanthoxylum bungeanum is extracted by supercritical carbon dioxide extraction.

[0053] Furthermore, the Zanthoxylum bungeanum is extracted by supercritical carbon dioxide extraction, which includes the following preparation process steps:

[0054] Step 1: Take the peppercorns and crush them, and extract them by supercritical carbon dioxide extraction, with an extraction temperature of 35-50°C, an extraction time of 1-4 hours, an extraction pressure of 16-28 MPa, and a carbon dioxide flow rate of 1.0-2.5 m 3 / h, to obtain the primary extract. Alternatively, the following process parameters can be used for the supercritical carbon dioxide extraction method to extract pepper: extraction temperature 40-60°C, extraction time 1-6 hours, extraction pressure 16-32MPa, carbon dioxide flow rate 50-100kg / h, to obtain the primary extract;

[0055] Step 2: Heat the primary and secondary extracts at 35-50℃ respectively, let them stand to remove moisture and precipitation.

[0056] Step 3: Mix the primary and secondary materials to obtain the Zanthoxylum bungeanum extract.

[0057] The pepper extract prepared by the present invention adopts a carbon dioxide supercritical extraction method. By controlling the extraction time and the carbon dioxide supercritical conversion temperature in the extraction process, the selectivity is better, and the ratio of amide components and volatile oil components in the obtained pepper oil amine is appropriate, so the pepper extract can be better used for treating atopic dermatitis, eczema, psoriasis and neurodermatitis.

[0058] More specifically, the present invention relates to the following technical solutions:

[0059] The invention discloses an application of zanthoxylum bungeanum in preparing medicines for treating atopic dermatitis, eczema, psoriasis and neurodermatitis.

[0060] Preferably, the Zanthoxylum bungeanum is prepared by supercritical extraction technology.

[0061] Preferably, the supercritical extraction technology preparation refers to: adding the pepper raw material into an extraction tank, and using carbon dioxide for supercritical extraction; during the extraction process, the CO2 temperature is controlled between 40 and 50°C, or is optionally controlled between 40 and 60°C.

[0062] Preferably, the supercritical extraction technology preparation refers to: extracting the pepper raw material by supercritical carbon dioxide extraction, with an extraction temperature of 40-50°C, an extraction time of 2-4 hours, an extraction pressure of 20-28 MPa, and a carbon dioxide flow rate of 1.5-2.5 m3 / h. Alternatively, the process parameters can be controlled as follows: 45-60°C, extraction time 1-6 hours, extraction pressure 16-32MPa, carbon dioxide flow rate 50-100kg / h.

[0063] Preferably, the Zanthoxylum bungeanum oleylamine is a Zanthoxylum bungeanum extract containing zanthoxylum bungeanum amide components and Zanthoxylum bungeanum volatile oil.

[0064] Preferably, the medicament is an external preparation.

[0065] Preferably, the drug is any one of a tincture, liniment, ointment, lotion, emulsion, film coating, gel, paste, transdermal patch, spray, and aerosol.

[0066] Preferably, the drug is a liquid preparation.

[0067] Preferably, the drug contains pharmaceutically acceptable excipients.

[0068] Compared with the prior art, the present invention has the following beneficial effects:

[0069] 1. The Zanthoxylum bungeanum extract of the present invention is used for the preparation of medicines, especially for the treatment of postherpetic pain (PHN). The amide components and volatile oil components in Zanthoxylum bungeanum oleamine are coordinated to achieve the effect of local medication to specifically inhibit PHN. The main component is a pure natural extract, which is easily accepted by patients, improves the compliance rate of doctors' orders, and plays a better therapeutic role.

[0070] 2. The pharmaceutical application of the Zanthoxylum bungeanum extract of the present invention is safer to use than the local anesthetics and capsaicin in the prior art, and has little impact on daily life.

[0071] 3. The Zanthoxylum bungeanum extract of the present invention contains ingredients such as prickly ash amide (hydroxy-α-sanshool and hydroxy-β-sanshool, etc.) and volatile oils (limonene, linalool), among which prickly ash amide can inhibit the Nav1.7 channel of neuronal endings, inhibit the influx of Na ions and block the transmission of pain, and can protect neurons and tactile receptors; combined with the prickly ash volatile oil ingredients, it further synergistically enhances the analgesic and transdermal penetration effects, and effectively alleviates the unbearable pain caused by the damage of the patient's nerve cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0072] Figure 1 This is the characteristic spectrum of Zanthoxylum bungeanum extract.

[0073] Figure 2 This is the liquid chromatogram of Zanthoxylum bungeanum extract.

[0074] Figure 3 These are photos comparing the recovery of the skin on the back of mice. DETAILED DESCRIPTION

[0075] The present invention is further described in detail below in conjunction with test examples and specific implementation methods. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following embodiments, and all technologies realized based on the content of the present invention belong to the scope of the present invention.

[0076] Example 1

[0077] Preparation of Zanthoxylum bungeanum extract

[0078] S1. Weigh 1000 g of pepper and fully dry it until the moisture content is less than 12%.

[0079] S2. Crush the peppercorns and extract them by supercritical carbon dioxide extraction at a temperature of 35°C, a time of 4 hours, an extraction pressure of 16 MPa, and a carbon dioxide flow rate of 1.0 m 3 / h to obtain the initial extract.

[0080] S3, step 2: heat the primary and secondary extracts to 30-35°C respectively, let stand, remove water and precipitate,

[0081] S4, step three: mixing the primary and secondary materials to obtain the Zanthoxylum bungeanum extract.

[0082] Example 2

[0083] Preparation of Zanthoxylum bungeanum extract

[0084] S1. Weigh 1000 g of pepper and fully dry it until the moisture content is less than 12%.

[0085] S2. Crush the peppercorns and extract them by supercritical carbon dioxide extraction at a temperature of 40°C, for 2 hours, at a pressure of 20 MPa, and at a carbon dioxide flow rate of 1.5 m 3 / h to obtain the initial extract.

[0086] S3, step 2: heat the primary and secondary extracts to 35-40°C respectively, let stand, remove water and precipitate,

[0087] S4, step three: mixing the primary and secondary materials to obtain the Zanthoxylum bungeanum extract.

[0088] Example 3

[0089] Preparation of Zanthoxylum bungeanum extract

[0090] S1. Weigh 1000 g of pepper and fully dry it until the moisture content is less than 12%.

[0091] S2. Crush the peppercorns and extract them by supercritical carbon dioxide extraction at a temperature of 45°C, for 3 hours, at a pressure of 24 MPa, and at a carbon dioxide flow rate of 2.0 m 3 / h to obtain the initial extract.

[0092] S3, step 2: heat the primary and secondary extracts to 35-40°C respectively, let stand, remove water and precipitate,

[0093] S4, step three: mixing the primary and secondary materials to obtain the Zanthoxylum bungeanum extract.

[0094] Example 4

[0095] Preparation of Zanthoxylum bungeanum extract

[0096] S1. Weigh 1000 g of pepper and fully dry it until the moisture content is less than 12%.

[0097] S2. Crush the peppercorns and extract them by supercritical carbon dioxide extraction at a temperature of 50°C, an extraction time of 1 hour, an extraction pressure of 28 MPa, and a carbon dioxide flow rate of 2.5 m 3 / h to obtain the initial extract.

[0098] S3, step 2: heat the primary and secondary extracts to 30-35°C respectively, let stand, remove water and precipitate,

[0099] S4, step three: mixing the primary and secondary materials to obtain the Zanthoxylum bungeanum extract.

[0100] Example 5

[0101] Preparation of Zanthoxylum bungeanum extract

[0102] S1. Weigh 1000 g of pepper and fully dry it until the moisture content is less than 12%.

[0103] S2. Crush the peppercorns and extract them by supercritical carbon dioxide extraction at a temperature of 60° C., an extraction time of 4 hours, an extraction pressure of 32 MPa, and a carbon dioxide flow rate of 90 kg / h to obtain a primary extract.

[0104] S3, step 2: heat the primary and secondary extracts to 30-50°C respectively, let stand, remove water and precipitate,

[0105] S4, step three: mixing the primary and secondary materials to obtain the Zanthoxylum bungeanum extract.

[0106] Example 6

[0107] Preparation of Zanthoxylum bungeanum extract

[0108] S1. Weigh 1000 g of pepper and fully dry it until the moisture content is less than 12%.

[0109] S2. Crush the peppercorns and extract them by supercritical carbon dioxide extraction at a temperature of 45° C., for 2 hours, at a pressure of 28 MPa, and at a carbon dioxide flow rate of 60 kg / h to obtain a primary extract.

[0110] S3, step 2: heat the primary and secondary extracts to 35-50°C respectively, let stand, remove water and precipitate,

[0111] S4, step three: mixing the primary and secondary materials to obtain the Zanthoxylum bungeanum extract.

[0112] Example 7

[0113] Preparation of Zanthoxylum bungeanum extract

[0114] S1. Weigh 1000 g of pepper and fully dry it until the moisture content is less than 12%.

[0115] S2. Crush the peppercorns and extract them by supercritical carbon dioxide extraction at a temperature of 50° C., for 5 hours, at a pressure of 28 MPa, and at a carbon dioxide flow rate of 90 kg / h to obtain a primary extract.

[0116] S3, step 2: heat the primary and secondary extracts to 35-50°C respectively, let stand, remove water and precipitate,

[0117] S4, step three: mixing the primary and secondary materials to obtain the Zanthoxylum bungeanum extract.

[0118] Example 8

[0119] Preparation of Zanthoxylum bungeanum extract

[0120] S1. Weigh 1000 g of pepper and fully dry it until the moisture content is less than 12%.

[0121] S2. Crush the pepper and extract it by supercritical carbon dioxide extraction at a temperature of 55° C., an extraction time of 6 hours, an extraction pressure of 24 MPa, and a carbon dioxide flow rate of 100 kg / h to obtain a primary extract.

[0122] S3, step 2: heat the primary and secondary extracts to 30-50°C respectively, let stand, remove water and precipitate,

[0123] S4, step three: mixing the primary and secondary materials to obtain the Zanthoxylum bungeanum extract.

[0124] Example 9

[0125] Characterization Test of Zanthoxylum bungeanum Extract

[0126] 1) Volatile oil content test

[0127] The Zanthoxylum bungeanum extract prepared by the carbon dioxide supercritical extraction method of Example 1 was tested for volatile oil components according to the method A of the volatile oil determination method 2204 of the Chinese Pharmacopoeia.

[0128] Take about 3g of the sample, weigh it (accurate to 0.01g), put it in a flask, add 200mL of water and a few glass beads, shake and mix, and connect the volatile oil analyzer and the reflux condenser. Add water from the upper end of the condenser until it fills the scale part of the volatile oil analyzer and overflows into the flask. Put it in an electric heating mantle and slowly heat it to boiling, and keep it at a slight boil for about 5 hours until the amount of oil in the analyzer no longer increases, stop heating, let it stand for a while, open the piston at the lower end of the analyzer, and slowly release the water until the upper end of the oil layer reaches 5mm above the scale 0 line. Let it stand for more than 1 hour, then open the piston to let the oil layer drop until its upper end is just flush with the scale 0 line, read the amount of volatile oil, and calculate the content (%) of volatile oil in the sample.

[0129] 2) Amide content test

[0130] The total amides of Zanthoxylum bungeanum extract were tested by ultraviolet spectrophotometry according to General Chapter 0401 of the Chinese Pharmacopoeia.

[0131] Preparation of reference solution: Take an appropriate amount of hydroxy-α-sanshool and add anhydrous ethanol to make a solution containing 0.1 mg of hydroxy-α-sanshool per 1 mL to obtain the reference solution.

[0132] Preparation of standard curve: accurately measure an appropriate amount of the above-mentioned hydroxy-α-sanshool reference solution, use anhydrous ethanol to prepare a standard series of solutions with a concentration of 0.2-10.0 μg / mL (prepare and use immediately, the concentration range can be appropriately adjusted according to the absorbance value), measure the absorbance value at a wavelength of 270 nm using an ultraviolet spectrophotometer, use anhydrous ethanol for blank correction, and draw a standard curve with the hydroxy-α-sanshool concentration as the horizontal coordinate and the absorbance value as the vertical coordinate.

[0133] Determination method: Take about 30 mg of the mixed sample, weigh it accurately, put it in a stoppered conical bottle, accurately add 25 mL of anhydrous ethanol, seal it, weigh it, ultrasonically treat it (power 500 W, frequency 40 kHz) for 15 minutes, let it cool, weigh it again, make up the lost weight with anhydrous ethanol, and shake it well. Accurately measure 50 μL of the solution, put it in a 10 mL volumetric bottle, add anhydrous ethanol to the scale, shake it well, and use it as the test solution. The absorbance value is measured at a wavelength of 270 nm by an ultraviolet spectrophotometer, and the concentration of hydroxy-α-sanshool in the test solution is read from the standard curve and calculated.

[0134] The test results are shown in the following table.

[0135] Table 1 Test results of Zanthoxylum bungeanum extract

[0136] Extract 1 Extract 2 batch number C6-0230-221007G C6-0230-220521G Total amides (%, g / g) 57.5 60.3 Volatile oil (%, mL / g) 28.0 29.2 Hydroxy-α-sanshool / Hydroxy-β-sanshool (%, g / g)* 47.3 47.7 Limonene / linalool (%, g / g) 17.3 13.4

[0137] *Note: For the determination method of the contents of hydroxy-α-sanshool / hydroxy-β-sanshool (%, g / g) and limonene / linalool (%, g / g) in the above table, refer to the gas chromatography and high performance liquid chromatography of Zanthoxylum bungeanum extract described later (see Example 10 Quality Control of Zanthoxylum bungeanum extract for details).

[0138] It can be seen that Zanthoxylum bungeanum is a CO2 supercritical extract of Zanthoxylum bungeanum, and is named Zanthoxylum bungeanum because it mainly contains amides and volatile oil components. Therefore, Zanthoxylum bungeanum can be preferably used as an active ingredient for preparing medicines, thereby obtaining medicines with the effect of alleviating skin tactile / sensory hypersensitivity.

[0139] Example 10

[0140] Quality Control of Zanthoxylum bungeanum Extract

[0141] In terms of quality control, the present invention simultaneously establishes the characteristic spectra of the amide and volatile oil parts for the first time, and determines the contents of volatile oil and total amide in the extract. In addition, the contents of limonene and linalool in the volatile oil and the contents of hydroxy-α-sanshool and hydroxy-β-sanshool in the amide are determined.

[0142] The Zanthoxylum bungeanum extract prepared in Example 1 was characterized by the following specific method:

[0143] 2.1 Gas chromatography characteristic spectrum (General Chapter 0521 of the Chinese Pharmacopoeia)

[0144] Chromatographic conditions and system suitability test

[0145] Capillary column with polyethylene glycol as stationary phase (column length 30m, inner diameter 0.25mm, film thickness 0.25μm); column temperature is programmed: initial temperature is 50℃, maintained for 4 minutes, then heated to 160℃ at a rate of 8℃ per minute, then heated to 240℃ at a rate of 10℃ per minute, maintained for 2 minutes. Inlet temperature 250℃℃; detector temperature 250℃℃; split injection, split ratio 30:1; carrier gas is nitrogen, flow rate is 1.2mL per minute. The theoretical plate number calculated based on the limonene peak should not be less than 2000000.

[0146] Preparation of reference solution

[0147] Take appropriate amounts of limonene, linalool, linalyl acetate, 4-terpineol, α-terpineol, L-carvone, eucalyptol, and myrcene reference substances, accurately weigh them, and add ethyl acetate to prepare a solution containing 0.3 mg of limonene, 0.1 mg of linalool, 50 μg of linalyl acetate, 30 μg of 4-terpineol, 15 μg of α-terpineol, 40 μg of L-carvone, 40 μg of eucalyptol, and 25 μg of myrcene as the reference solution.

[0148] Preparation of test solution

[0149] Take about 40 mg of the mixed sample, accurately weigh it, place it in a stoppered conical flask, accurately add 50 mL of ethyl acetate, stopper it, weigh it, ultrasonically treat it (power 500 W, frequency 40 kHz) for 20 minutes, cool it, weigh it again, make up the lost weight with ethyl acetate, shake it well, filter it, and you will get the test solution.

[0150] Determination method

[0151] Accurately pipette 1 μL of the reference solution and the test solution respectively, inject into the gas chromatograph, and measure to obtain the result.

[0152] The reference solution is measured to obtain a gas phase reference characteristic spectrum, such as Figure 1 As shown in the figure, peak 1: myrcene, peak 2 (S): limonene, peak 3: linalool, peak 4: linalool, peak 5: linalyl acetate, peak 6: 4-terpineol, peak 7: L-carvone.

[0153] The test solution is measured to obtain the test liquid phase characteristic spectrum, such as Figure 2 As shown in the figure, 7 characteristic peaks should be presented. The peak corresponding to the limonene reference peak is the S peak. The relative retention time of each characteristic peak and the S peak should be within ±5% of the specified value. The specified values ​​of retention time are: 0.92 (peak 1), 1.00 [peak 2 (S)], 1.02 (peak 3), 1.70 (peak 4), 1.72 (peak 5), 1.81 (peak 6), and 2.04 (peak 7).

[0154] 2.2 Characteristic spectrum of high performance liquid chromatography (General Chapter 0512 of the Chinese Pharmacopoeia)

[0155] Chromatographic conditions and system suitability test

[0156] Octadecylsilane bonded silica gel is used as filler (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); water is used as mobile phase A, methanol is used as mobile phase B, and gradient elution is performed according to the provisions in the following table; column temperature is 35°C; flow rate is 1 mL / min; detection wavelength is 270 nm. The theoretical plate number calculated based on hydroxy-α-sanshool should not be less than 50,000.

[0157] Table 2 Gradient elution program

[0158]

[0159] Preparation of reference solution

[0160] Take appropriate amount of hydroxy-ε-sanshool, hydroxy-α-sanshool, hydroxy-β-sanshool, hydroxy-γ-sanshool, and γ-sanshool reference substances, weigh them accurately, and add anhydrous ethanol to make solutions containing 5 μg of hydroxy-ε-sanshool, 0.5 mg of hydroxy-α-sanshool, 5 μg of hydroxy-β-sanshool, 0.25 mg of hydroxy-γ-sanshool, and 50 μg of γ-sanshool per 1 mL as reference solutions.

[0161] Preparation of test solution

[0162] Take about 30 mg of the mixed sample, weigh it accurately, put it in a stoppered conical bottle, accurately add 25 mL of anhydrous ethanol, stopper it, weigh it, ultrasonically treat it (power 500 W, frequency 40 kHz) for 15 minutes, let it cool, weigh it again, make up the lost weight with anhydrous ethanol, and shake it well. Accurately measure 500 μL of the solution, put it in a 10 mL volumetric bottle, add anhydrous ethanol to the scale, shake it well, and use it as the test solution.

[0163] Determination method

[0164] Accurately pipette 5 μL of the reference solution and the test solution respectively, inject into the liquid chromatograph, and measure to obtain the result.

[0165] The reference solution is measured by liquid chromatography to obtain a control characteristic spectrum, such as Figure 1 In the figure, peak 1: hydroxy-ε-sanshool, peak 2 (S): hydroxy-α-sanshool, peak 3: hydroxy-β-sanshool, peak 4: hydroxy-γ-sanshool, peak 5: γ-sanshool.

[0166] The test solution is measured by liquid chromatography to obtain the test sample characteristic spectrum, such as Figure 2 As shown. There should be 5 characteristic peaks in the figure. The peak corresponding to the hydroxy-α-sanshool reference peak is the S peak. The relative retention time of each characteristic peak and the S peak should be within ±10% of the specified value. The specified values ​​of retention time are: 0.60 (peak 1), 1.00 [peak 2 (S peak)], 0.55 (peak 3), 4.47 (peak 4), 7.75 (peak 5).

[0167] Exploration Experiment Example 1

[0168] Preparation of Zanthoxylum bungeanum volatile oil

[0169] S1. Weigh 1000 g of pepper and fully dry it until the moisture content is less than 12%.

[0170] S2. Take coarse powder of Zanthoxylum bungeanum, extract it by steam distillation for 5 hours, let it stand to remove water, and obtain volatile oil of Zanthoxylum bungeanum.

[0171] Exploration Experiment Example 2

[0172] Preparation of Zanthoxylum bungeanum water extract

[0173] S1. Weigh 1000 g of pepper and fully dry it until the moisture content is less than 12%.

[0174] S2. Take coarse powder of Zanthoxylum bungeanum, add 10 times the amount of water, boil, extract for 1 hour, filter, add 8 times the amount of water to the residue, decoct and extract for 1 hour, combine the extracts to obtain a water extract.

[0175] S3. Using a rotary evaporator, vacuum decompression, concentration and drying are performed to obtain the Zanthoxylum bungeanum water extract.

[0176] Exploration Experiment Example 3

[0177] Preparation of Zanthoxylum bungeanum alcohol extract

[0178] S1. Weigh 1000 g of pepper and fully dry it until the moisture content is less than 12%.

[0179] S2. Take coarse powder of Zanthoxylum bungeanum, add 10 times the amount of 95% ethanol, reflux extract for 1 hour, filter, add 8 times the amount of 95% ethanol to the residue, reflux extract for another 1 hour, combine the extracts to obtain an alcohol extract.

[0180] S3. Using a rotary evaporator, vacuum decompression concentration and drying are performed to obtain the Zanthoxylum bungeanum alcohol extract.

[0181] Exploration Experiment Example 4

[0182] Preparation of Zanthoxylum bungeanum ether extract

[0183] S1. Weigh 1000 g of pepper and fully dry it until the moisture content is less than 12%.

[0184] S2. Take coarse powder of Zanthoxylum bungeanum, add 10 times amount of ether, reflux extract for 1 hour, filter, add 8 times amount of ether to the residue, reflux extract for another 1 hour, combine the extracts to obtain ether extract.

[0185] S3. Using a rotary evaporator, vacuum decompression, concentration and drying are performed to obtain the Zanthoxylum bungeanum ether extract.

[0186] Exploration Experiment Example 5

[0187] Preparation of pepper amide extracts

[0188] S1. Weigh 1000 g of pepper and fully dry it until the moisture content is less than 12%.

[0189] S2. Take the coarse powder of Sichuan pepper, add 10 times the amount of methanol, reflux extract for 1 hour, filter, add 8 times the amount of methanol to the residue, reflux extract for another 1 hour, combine the extracts, and concentrate them with a rotary evaporator to obtain a crude amide extract. The crude extract is passed through a preparative silica gel column, using methanol: water (10:1) as the eluent, eluting, collecting the elution segment containing the amide component, and obtaining an eluent.

[0190] S3. Use a rotary evaporator to concentrate under vacuum and dry to obtain the prickly ash amide extract.

[0191] Experimental Example 1

[0192] Effect of Zanthoxylum bungeanum on allylisothiocyanate (AITC)-induced neuroinflammation in mice The role of mechanical pain threshold (animal experiment)

[0193] The mechanism of PHN caused by herpes zoster is relatively complex. Studies have found that it may be related to the loss of nerve fiber damage caused by long-term viral chronic infection, increased activity of sensory afferent fibers, abnormal discharges and central overexcitation caused by them. When AITC is applied to the soles of rats' feet, it can cause local inflammation of nerve endings and neuropathic pain, thus simulating the pain and pain hypersensitivity caused by PHN.

[0194] Animal grouping and administration: Mice were divided into 4 groups (10 in each group): blank control group, model control group, positive drug control group (5% compound lidocaine cream), 5% Zanthoxylum group (Zanthoxylum group prepared in Example 1, dissolved in dimethylformamide (DMF)), 5% volatile oil group, 5% amide group (DMF dissolved), 5% water extract group (DMF dissolved), 5% alcohol extract group (DMF dissolved), 5% ether extract group (DMF dissolved), 5% hydroxy-α-sanshool group (DMF dissolved), 5% hydroxy-β-sanshool group (DMF dissolved). The drug concentrations of the above-mentioned test groups are all weight percentages by weight. Except for the blank control group, 10% AITC (5 μL) was applied to the soles of the mice, and the positive drug and Zanthoxylum group (a full layer of drug was applied to the soles) were immediately applied after application. After 40 minutes of administration, the mechanical pain threshold of each group of mice was determined using the Von-Frey acupuncture pain test kit. After administration, whether the mechanical pain threshold of mice can be prolonged was evaluated.

[0195] Specific method for index evaluation: mice were randomly placed in a cage with a wire mesh bottom of 1cm×1cm aperture. After the mice were adapted to the cage for 30 minutes, the soles of both feet of the mice were vertically stimulated with von Frey fiber filaments of different strengths for 1 to 3 seconds. The positive reaction was the lifting or licking of the feet, otherwise it was a negative reaction. The measurement was performed 5 times in a row, with an interval of 10 seconds each time. The positive result was recorded as a positive result if 3 out of 5 measurements showed a positive reaction. The average value of the left and right sides was the final mechanical foot withdrawal threshold of the mice.

[0196] Similar to the positive drug (lidocaine), local application of Zanthoxylum bungeanum group can significantly increase the mechanical pain threshold of mice with neuroinflammation, and the effect of the same dose of Zanthoxylum bungeanum is better than the simple addition of the application of volatile oil alone and the application of amide alone, with obvious synergistic effect, and the effect of Zanthoxylum bungeanum is significantly better than the water extract, alcohol extract and ether extract groups, as well as the hydroxy-α-sanshool group (DMF dissolved) and hydroxy-β-sanshool group (DMF dissolved), with obvious technical progress.

[0197] Table 3. Effects of Zanthoxylum bungeanum on mechanical pain threshold of mice with neuroinflammation induced by allyl isothiocyanate

[0198]

[0199] The experimental results show that the effect of the prickly ash amine prepared in Example 1 on improving the mechanical pain threshold of the neuroinflammatory mice is the most prominent, reaching 4.25±0.25 seconds, while the volatile oil and amide groups are 3.72±0.31 seconds and 3.91±0.18 seconds, respectively. That is, at the same dosage (5% dilution) ratio, prickly ash amine, as a complex containing volatile oil and amide, can exert a better antipruritic effect, which is better than applying volatile oil alone or amide alone, and has a synergistic effect.

[0200] Further comparison of ether, hydroxy-α-sanshool and hydroxy-β-sanshool shows that the effect of zanthoxylum bunge is also the best, which means that the effect can not be achieved by a single sanshool ingredient, but there is a more complex mechanism of action.

[0201] Experimental Example 2

[0202] Experimental study on the irritation of zanthoxylum oleylamine and capsaicin on mouse skin

[0203] Capsaicin has a certain effect in treating PHN, but long-term use can cause strong skin irritation. This study compared the skin irritation of the same dose of Zanthoxylum bungeanum extract and capsaicin on mice.

[0204] Animal grouping: Healthy rats were divided into 3 groups (10 in each group): blank control group, 5% capsaicin group (purity 98%, dissolved in DMF), 5% zanthoxylum group (zanthoxylum prepared in Example 1, dissolved in DMF), the drug concentrations of the above test groups were all weight percentage wt%.

[0205] The back of the rat was depilated with 8% barium sulfide with the spine as the boundary, and the experiment was carried out 24 hours later. The drug was tested on the right side of the depilated area of ​​the animal, 1 mL / animal, and DMF solvent was applied on the left side, 1 mL / animal, as the control group. The drug administration area was covered with gauze and plastic wrap, fixed with tape for 4 hours, and then the plastic wrap was removed, and the residual drug on the skin was washed off with saline. Once / day, continuous administration for 7 days, 24 hours after the last administration, the residual drug solution and solvent were washed off with warm water. Observe and record the presence or absence of edema and erythema on the local skin of each animal at 1h, 24h, 48h, and 72h after the removal of the test substance, and perform skin irritation reaction scores (Table 4), the average value of the stimulation test reaction = (total score of edema formation + total score of erythema formation) / total number of animals. Perform skin irritation intensity scoring. If obvious lesions are observed, skin pathological tissue examination is performed 72h after the last administration.

[0206] Table 4 Skin irritation response scoring criteria

[0207]

[0208] The skin irritation rating results all showed that (

[0209] ), under the same dosage, the irritation of Zanthoxylum bungeanum extract is significantly less than that of capsaicin. The above results show that Zanthoxylum bungeanum extract is expected to be used to treat postherpetic pain (PHN) with less side effects.

[0210] Table 5. Irritation of Zanthoxylum bungeanum and capsaicin to mouse skin

[0211]

[0212] Note: # P>0.05, * P<0.05.

[0213] Experimental Example 3

[0214] Interventional effects of fennel amine on tactile / sensory hypersensitivity induced by varicella-zoster virus in rats

[0215] Methods: Male SD rats weighing 200-250g were randomly divided into 11 groups (10 rats in each group), namely: blank control group, model control group, positive drug control group (5% compound lidocaine cream), 5% zanthoxylum group (zanthoxylum group prepared in Example 1, dissolved in DMF), 5% volatile oil group, 5% amide group (dissolved in DMF), 5% water extract group (dissolved in DMF), 5% alcohol extract group (dissolved in DMF), 5% ether extract group (dissolved in DMF), 5% hydroxy-α-sanshool group (dissolved in DMF), and 5% hydroxy-β-sanshool group (dissolved in DMF). The experimental reagents were prepared in the same way as in Experimental Example 1, and the drug concentrations of each test group were all weight percentages (wt%).

[0216] The animal model of chronic infection of varicella zoster virus was established according to the literature. Vero cells (African green monkey kidney cells) were first added to DMEM / F12 complete medium to prepare a cell suspension. 4 / cm 2 Inoculate in culture flasks, then infect Vero cells with the virus. After 2 days, collect the infected cells and prepare the cell suspension as the virus inoculation solution. Finally, 50 μL (which contains approximately 6×10 6 The inoculation solution of 100 infected cells (100 infected cells) was injected into the left toe of SD rats. The model was successfully established when the rats began to show abnormal mechanical pain threshold 3 days after virus infection. The drug was administered on the day of model establishment (a layer of drug was applied to the sole of the foot), and the mechanical pain threshold of rats in each group was measured before model establishment and 3, 7, 14, and 21 days after model establishment using the VonFrey acupuncture pain test kit. After drug administration, whether the mechanical pain threshold of rats could be prolonged was evaluated.

[0217] Specific method for index evaluation: Rats were randomly placed in a cage with a wire mesh bottom of 1cm×1cm aperture. After the rats were adapted to the cage for 30 minutes, the soles of both feet of the rats were vertically stimulated with von Frey fiber filaments of different strengths for 1 to 3 seconds. The presence of foot lifting or licking behavior was considered a positive reaction, otherwise it was considered a negative reaction. The measurement was performed 5 times in a row, with an interval of 10 seconds each time. A positive result was recorded if 3 out of 5 measurements showed a positive reaction, and the average value of the left and right sides was the final mechanical foot withdrawal threshold of the rats.

[0218] The experimental results are shown in Table 6. Similar to the positive drug (lidocaine), local application of prickly ash oil amine can significantly increase the pain threshold of rats induced by herpes zoster virus. This model is caused by chronic infection of herpes zoster virus, which is an animal model closer to the real pain and hypersensitivity of herpes zoster patients. The mechanical withdrawal threshold of rats in each group was measured 14 and 21 days after modeling, which can fully reflect the characteristics of post-herpetic pain, that is, the virus has been cleared and the inflammation has subsided. The symptoms at this time are mainly pain or pain / hypersensitivity caused by post-herpetic nerve damage.

[0219] Moreover, the effect of the same dose of Zanthoxylum bungeanum is better than that of the volatile oil group and the amide group alone, and has obvious synergistic effect. The effect of Zanthoxylum bungeanum is significantly better than that of the water extract, alcohol extract and ether extract groups, as well as the hydroxy-α-sanshool group (DMF dissolved) and hydroxy-β-sanshool group (DMF dissolved), and has obvious technical progress.

[0220] Table 6. Interventional effects of Zanthoxylum bungeanum on tactile / sensory hypersensitivity in rats induced by varicella zoster virus

[0221]

[0222] Note: # P>0.05, * P<0.05, ** P<0.01, *** P<0.001vs model group.

[0223] Experimental Example 4

[0224] The therapeutic effect of Zanthoxylum bungeanum on atopic dermatitis in mice induced by dinitrochlorobenzene (DNCB) (Animal experiment)

[0225] Animal grouping and dosing:

[0226] SPF male BALB / c mice, weighing (20±2)g, 6 weeks old, were divided into 11 groups (10 mice in each group): blank control group, model group, positive control group (mometasone furoate cream), 5% sanshoolamine group (dissolved in dimethylformamide (DMF)), 5% volatile oil group, 5% amide group (dissolved in DMF), 5% water extract group (dissolved in DMF), 5% ethanol extract group (dissolved in DMF), 5% ether extract group (dissolved in DMF), 5% hydroxy-α-sanshool group (dissolved in DMF), and 5% hydroxy-β-sanshool group (dissolved in DMF). Except for the blank control group, which was given blank matrix solution (acetone: olive oil = 9:3), the other groups were stimulated with DNCB solution on the back and right ear of mice to establish AD models.

[0227] 0d: depilatory cream was used to remove the hair on the back of the mice, with an area of ​​about 3×3 cm; 1-3d: 150 μL and 30 μL of 1% DNCB were applied to the depilated area on the back and the right ear, respectively, and the left ear was treated with blank matrix solution, once a day.

[0228] 4-6d: 150 μL and 30 μL of 0.5% DNCB were applied to the back skin and right ear of the mice, respectively, and the left ear was treated with an equal amount of blank matrix solution, once a day.

[0229] 7-21d: 0.5% DNCB 150μL and 30μL were applied to the back skin and right ear of mice, respectively, and the left ear was treated with an equal amount of blank matrix solution, once every two days. After 12h of stimulating the back and right ear skin of mice with DNCB solution each time, the positive drug control group and the experimental drug group were smeared with the prepared drug solution, and the blank control group and the model control group were smeared with an equal amount of DMF.

[0230] 22-28d: No DNCB solution stimulation was used, only PAC and ZBE were applied by smear.

[0231] Specific methods for indicator evaluation:

[0232] ① Scratching frequency statistics

[0233] On the 21st day of the experiment, the number of scratches of the mice within 10 minutes after the last sensitization with 0.5% DNCB was recorded. The judgment criteria were: if the mouse scratched the skin of the area stimulated by DNCB (i.e., the back and the right ear), it was counted as one scratch. If the scratch lasted for 3 seconds, it was counted as two scratches. The video was recorded for 10 minutes. Finally, the total number of continuous scratches of the mice within 10 minutes was counted based on the video.

[0234] ②Skin lesion score

[0235] On the 27th day of the experiment, the inflammation of the back skin and right ear of each group of mice after sensitization with DNCB was recorded and scored. The severity score of the lesion was used to evaluate the degree of dermatitis on the back skin of each group of mice: including erythema, hypertrophy, scaling, edema and epidermal exfoliation, and the scoring levels were: no symptoms: 0 points, mild symptoms: 1 point, moderate symptoms: 2 points, severe symptoms: 3 points. The dermatitis score was calculated as the sum of individual scores.

[0236] ③Ear thickness difference statistics

[0237] The ear thickness difference was calculated based on the measured thickness of the left and right ears of the mice, that is, ear thickness difference (mm) = DNCB-sensitized right ear thickness (mm) - non-sensitized left ear thickness (mm).

[0238] ④ELISA to detect serum IgE and skin IL-6, IL-13, IL-31, TNF-α levels

[0239] Place the whole blood sample at room temperature for 2 hours, centrifuge at 4°C, 1000Xg for 20 minutes, and take the supernatant. Take the back skin tissue of each group of mice, rinse the tissue with pre-cooled PBS, weigh it, cut the tissue into pieces and put it into a centrifuge tube. Add pre-cooled PBS (tissue: PBS = 1:9) and homogenize, ice bath for 30 minutes, and ultrasonically treat it 3 times during the period. Finally, centrifuge the homogenate at 4°C, 5000Xg for 10 minutes, and take the supernatant. According to the instructions of the ELISA kit, the levels of mouse serum IgE and skin IL-6, IL-13, IL-31, and TNF-α were measured.

[0240] Experimental results (see Table 7 and Table 8, Figure 3 ): Topical application of Zanthoxylum bungeanum can significantly improve the skin lesions and dermatitis status of atopic dermatitis model mice, reduce the number of scratching times, and reduce the expression level of related inflammatory factors in mice. The same dose of Zanthoxylum bungeanum is better than the group applying volatile oil alone and the group applying amide alone, with obvious synergistic effects. Zanthoxylum bungeanum is significantly better than the water extract, alcohol extract and ether extract groups, as well as the hydroxy-α-sanshool group and the hydroxy-β-sanshool group, with obvious technical progress.

[0241] Table 7 Effects of Zanthoxylum bungeanum on inflammation in mice with atopic dermatitis

[0242]

[0243] Note: # P>0.05, * P<0.05, ** P<0.01, *** P<0.001vs model group.

[0244] Controlled by the experimental results in the above table, through the atopic dermatitis mouse experiment, it can be seen that when using prickly ash amine, it can reduce the number of inflammatory scratching of mice to a greater extent than the volatile oil and amide test groups. Although the water extract group scratched more and was basically the same as the ether extract test group, the number of scratching in the pure volatile oil test group was also high. This may be because prickly ash amine reduces the number of scratching of mice mainly through the mechanism of improving dermatitis, which has multiple pathways, and is not a simple action principle of a single ingredient.

[0245] Table 8 Effect of Zanthoxylum bungeanum on inflammation in mice with atopic dermatitis

[0246]

[0247]

[0248] Note: # P>0.05, * P<0.05, ** P<0.01, *** P<0.001vs model group.

[0249] From the experimental results in the above table, it can be seen that zanthoxylum bungeanum can reduce the expression of related inflammatory factors in mice, and its effect is better than that of single-ingredient extracts or water extracts. This is consistent with the results of the aforementioned winding experiment, and also proves that zanthoxylum bungeanum has a synergistic effect, rather than a single active ingredient and a single pathway effect.

[0250] Experimental Example 5

[0251] Therapeutic effect of zanthoxylum bungeanum on rats with neurodermatitis

[0252] Methods: Male SD rats weighing 200-250g were randomly divided into 11 groups (6 rats in each group), namely: blank control group, model control group, positive drug control group (halometasone cream), 5% zanthoxylum amine group (DMF dissolved), 5% volatile oil group, 5% amide group (DMF dissolved), 5% water extract group (DMF dissolved), 5% alcohol extract group (DMF dissolved), 5% ether extract group (DMF dissolved), 5% hydroxy-α-sanshool group (DMF dissolved), and 5% hydroxy-β-sanshool group (DMF dissolved).

[0253] Local skin friction irritation in rats:

[0254] After the rats were fixed, a 4-cm diameter area was selected on their backs. The hair was removed with a depilatory agent until the skin was exposed. The skin was rubbed with a homemade electric friction instrument until local congestion and bleeding points appeared. Each time was 5 minutes, once a day. Chronic mild unpredictable stress: The stimulation methods used included: 24-hour fasting, water deprivation, overnight lighting, thermal stimulation (5 minutes in a 45℃ electric thermostat), etc. In order to prevent the rats from adapting, more than one stimulation method was randomly given every day so that the rats could not expect the occurrence of stimulation. The modeling method combining local skin friction stimulation with chronic mild unpredictable stress lasted for 8 weeks, and the drug was continuously administered for 4 weeks after modeling. The serum was separated and prepared, and the levels of IL-1β, IL-4, IL-6 and TNF-α in the serum were detected by ELISA.

[0255] The experimental results show (see Table 9) that 5% sanshoolamine can effectively reduce the content of IL-1β, IL-4, IL-6 and TNF-α in serum and relieve the inflammatory response of rats. The same dose of sanshoolamine is better than the volatile oil and amide groups alone, with obvious synergistic effects, and the effect of sanshoolamine is significantly better than the water extract, alcohol extract and ether extract groups, as well as the hydroxy-α-sanshool group (DMF dissolved) and the hydroxy-β-sanshool group (DMF dissolved), with obvious technical progress.

[0256] Table 9 Effect of Zanthoxylum bungeanum on inflammation in rats with neurodermatitis

[0257]

[0258] Note: # P>0.05, * P<0.05, ** P<0.01, *** P<0.001vs model group.

Claims

1. Use of a Zanthoxylum bungeanum extract in the preparation of a drug for treating postherpetic pain, herpes zoster pain, atopic dermatitis, allergic dermatitis, senile skin pruritus and neurodermatitis.

2. The use according to claim 1, characterized in that: The invention discloses an application of Zanthoxylum bungeanum extract in preparing a medicine for treating itching and pain caused by at least one of post-herpetic pain, herpes zoster pain, atopic dermatitis, allergic dermatitis, neurodermatitis or senile skin pruritus.

3. Use of a Zanthoxylum bungeanum extract in the preparation of a drug for treating itching and pain caused by at least one of eczema, psoriasis or diabetic foot.

4. The use according to claim 1, characterized in that: The Zanthoxylum bungeanum extract is prepared by supercritical extraction technology.

5. The use according to claim 4, characterized in that: The supercritical extraction technology preparation refers to: adding the pepper raw material into the extraction tank and using carbon dioxide for supercritical extraction; during the extraction process, the CO2 temperature is controlled between 30 and 60 °C.

6. The use according to claim 5, characterized in that: The supercritical extraction technology preparation refers to: taking the pepper raw material and extracting it by supercritical carbon dioxide extraction, the extraction temperature is 30~60℃, the extraction time is 2~4 hours, the extraction pressure is 15~30MPa, and the carbon dioxide flow rate is 1.0~3m 3 / h.

7. The use according to claim 1, characterized in that: The postherpetic pain includes one or more of pain, allodynia, and allodynia, with or without itching.

8. The use according to claim 1, characterized in that: The Zanthoxylum bungeanum extract is Zanthoxylum bungeanum oleylamine containing Zanthoxylum bungeanum amide components and Zanthoxylum bungeanum volatile oil.

9. The use according to claim 1, characterized in that: The medicine is an external preparation.

10. The use according to claim 9, characterized in that: The medicine is any one of tincture, liniment, ointment, lotion, emulsion, film coating, gel, paste, transdermal patch, spray and aerosol.

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