CN115463186B - Mongolian medicine emplastrum, preparation method and application thereof - Google Patents
Mongolian medicine emplastrum, preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to the technical field of Mongolian medicine preparations, in particular to a Mongolian medicine emplastrum, a preparation method and application thereof. The invention provides a Mongolian medicine emplastrum, which comprises a emplastrum and a wetting solvent; the patch ointment is prepared from the following raw materials in parts by weight: 60-80 parts of raw kusnezoff monkshood root, 50-60 parts of rhizoma acori graminei, 50-60 parts of elecampane, 30-40 parts of liquidambar formosana hance, 50-60 parts of cassia seed and 50-60 parts of abutilon seed; the wetting solvent comprises the following components in parts by weight: 10-20 parts of peppermint oil, 15-25 parts of camphor, 10-20 parts of vanillyl butyl ether, 10-20 parts of methyl salicylate and 20-30 parts of light liquid paraffin. The Mongolian medicine emplastrum provided by the invention has no skin irritation, basically no sensitization reaction, good safety, better anti-inflammatory and analgesic effects, and better curative effect, and can be used for treating migraine.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicine preparations, in particular to a Mongolian medicine emplastrum, a preparation method and application thereof.
Background
The rheumatism is common and frequent, belongs to the category of Chinese medical arthralgia, and is mainly caused by invasion of four exogenous pathogens of wind, cold, dampness and heat into the body, and is most remarkable in heart and joint involvement. Rheumatic arthritis is one of the main manifestations of rheumatic fever, with 91.7% of adult affected joints in patients and 55.7% in children. About 50% of patients' conditions may develop to the heart. The disease incidence rate is up to more than 10%, the treatment is not in time, the disability rate is over 50%, and even the life is endangered.
For many years, western medicine is mainly used for treating rheumatism and rheumatoid diseases, and western medicine is mainly used for diminishing inflammation and easing pain, can only treat symptoms and cannot treat root causes, forms medicine dependence, causes complications to increase day by day, causes greater and greater damage to renal functions, destroys physical and mental health of people, and shortens the life of people. Practice proves that the traditional treatment method is difficult to radically cure rheumatism and rheumatoid.
The expert considers that the treatment principle is based on the concept of unity, and the traditional Chinese medicine is used for carrying out overall treatment based on syndrome differentiation, so that not only is the toxic and side effects greatly reduced, but also the toxic and side effects caused by taking western medicines for a long time are solved, and the traditional Chinese medicine has effects on various patients suffering from rheumatism and rheumatoid diseases which are not cured after long-term treatment.
In addition, the results of "Chinese headache epidemiological investigation" published by the Chinese medical society of pain university at a later date indicate that the incidence of primary headache is 23.8% in the 18-65 year old population in China, and nearly 1/4 Chinese (approximately more than 3 hundred million people calculated on the basis of the total population of the 5 th national census statistics) suffers from headache. Among them, the most common tension headache and migraine are 10.77% and 9.3%, respectively, and each city patient spends 1098.08 yuan per year on average for treating headache, and the investigation involves 5041 people in 6 large areas of China.
Headache is largely classified into primary and secondary. Secondary headaches are often caused by disease; primary headaches mainly include tension headaches and migraine headaches. At present, the incidence of headache in China is on an ascending trend, and the headache is not a occasional disease but a common chronic disease which is easy to repeatedly attack. There are two main causes of increased headache population. Firstly, people with increased stress and emotional problems such as anxiety, depression and the like are more and more, and patients with tension headache are more and more entangled, and headache is caused by stay up, long-time table work, irregular sleep and the like; secondly, compared with the prior art, the patient is more willing to go to the hospital to search for the cause.
Headache is a very dangerous, especially migraine, hazard. The world health organization even identifies severe migraine as one of the most disabling chronic diseases. Migraine has the characteristic of repeated attacks, can increase the complication risk, can increase the ischemic stroke risk by 2.16 times and can increase the abnormal brain white matter risk by 3.9 times. Surveys also show that 85% of the population is therefore unable to concentrate on work.
Compared with the severity of the disease, people pay much less attention to the headache, more than 60% of people can eat the analgesic without being able to eat the analgesic, which is the biggest misarea of people treating the headache. Headache causes are very complex and, in cases of ambiguous causes, intense headache can exacerbate symptoms.
The treatment of migraine comprises traditional Chinese medicine acupuncture, nonspecific treatment medicines, specific treatment and the like, and the traditional Chinese medicine acupuncture therapy can have a certain effect on relieving pain symptoms of migraine, but is difficult to treat and remove roots. Non-steroidal anti-inflammatory drugs (NSAIDs), such as acetaminophen, aspirin and ibuprofen, have significant gastrointestinal side effects and are not suitable for long-term administration. Barbiturates sedatives and opioids are easy to addict and are only suitable for other patients with severe migraine who are not effective in treatment. Specific therapeutic agent: ergot formulations are 5-HT receptor non-selective agonists, and common drugs are ergotamine and dihydroergotamine, which can terminate the acute onset of migraine. Triptan drugs may exert analgesic effects by constricting cerebral vessels, inhibiting nerve pain transmission in peripheral nerves and secondary neurons of the "trigeminal cervical complex". However, these drugs still have serious adverse reactions and are not suitable for long-term administration.
Disclosure of Invention
In view of the above, the technical problem to be solved by the invention is to provide a Mongolian medicine emplastrum, a preparation method and application thereof, wherein the Mongolian medicine emplastrum has no skin irritation, basically no sensitization reaction, good safety and better anti-inflammatory and analgesic effects, and can be used for treating migraine.
The Mongolian medicine emplastrum provided by the invention comprises a emplastrum and a wetting solvent; the patch ointment is prepared from the following raw materials: radix Aconiti Kusnezoffii (RadixAconiti Kusnezoffii), rhizoma Acori Calami (Acorus calamus), radix aucklandiae (Aucklandiae Radix), resina Liquidambaris (RESINA LIQUIDAMBARIS), semen Cassiae (Catsia tora Linn) and semen Abutili (Chingma Abutilon Seed). Wherein, the radix aconiti kusnezoffii (Radix Aconiti Kusnezoffii) has certain peculiar effects of dispelling cold and relieving pain, dispelling cold of channels and collaterals, dispelling cold of viscera, rheumatoid arthritis and heart and abdomen pain; rhizoma Acori Calami (Acorus calamus) has effects of eliminating phlegm, inducing resuscitation, invigorating spleen, and promoting diuresis. For epilepsy, palpitation, amnesia, mental confusion, damp-stagnation, distention and fullness, diarrhea, dysentery, rheumatalgia, carbuncle, swelling and scabies. Radix aucklandiae (Aucklandiae Radix) has effects of activating qi-flowing, relieving pain, warming spleen and stomach, relieving diarrhea with astringents, and can be used for treating chest and hypochondrium distending pain, abdominal distention, etc. The resina Liquidambaris (RESINA LIQUIDA MBARIS) has effects of promoting blood circulation, relieving pain, removing toxic substances, promoting granulation, and cooling blood, and can be used for treating traumatic injury, carbuncle, swelling and pain, hematemesis, traumatic hemorrhage, etc. Semen Cassiae (Catsia tora Linn) has effects of removing liver fire, improving eyesight, and relaxing bowels. It is used for treating hypertension, headache, vertigo, acute conjunctivitis, corneal ulcer, glaucoma, carbuncle, furuncle, and pyocutaneous disease. Abutilon (Chingma Abutilon Seed), also known as Abutilon, has effects of clearing heat, promoting diuresis, removing toxic substance and removing nebula, and is used for relieving carbuncle and nebula membrane. The raw materials of the patch plaster are extracted by a modern process to prepare a patch plaster which has better anti-inflammatory and analgesic effects and can be used for treating rheumatic arthritis, pain in the waist, soreness of the legs, migraine and the like.
The Mongolian medicine emplastrum provided by the invention comprises a emplastrum and a wetting solvent;
the patch ointment is prepared from the following raw materials in parts by weight: 60-80 parts of raw kusnezoff monkshood root, 50-60 parts of rhizoma acori graminei, 50-60 parts of elecampane, 30-40 parts of liquidambar formosana hance, 50-60 parts of cassia seed and 50-60 parts of abutilon seed;
the wetting solvent comprises the following components in parts by weight: 10-20 parts of peppermint oil, 15-25 parts of camphor, 10-20 parts of vanillyl butyl ether, 10-20 parts of methyl salicylate and 20-30 parts of light liquid paraffin.
In certain embodiments of the invention, the patch ointment and wetting solvent are stored separately and are mixed only when used.
Preferably, the patch ointment is prepared from the following raw materials in parts by weight:
75 parts of raw kusnezoff monkshood root, 52.5 parts of rhizoma acori graminei, 52.5 parts of costustoot, 32.5 parts of liquidambar formosana hance, 52.5 parts of cassia seed and 52.5 parts of abutilon seed;
the wetting solvent comprises the following components in parts by weight: 12 parts of peppermint oil, 20 parts of camphor, 10 parts of vanillyl butyl ether, 10 parts of methyl salicylate and 27 parts of light liquid paraffin.
In certain embodiments of the invention, the ratio of the patch ointment to the wetting solvent is 0.5 to 1.5g: 1.5-2.5 mL; specifically, it may be 1g:2mL.
The invention also provides a preparation method of the Mongolian medicine emplastrum, which comprises the following steps of:
mixing radix Aconiti Kusnezoffii, rhizoma Acori Calami, radix aucklandiae, resina Liquidambaris, semen Cassiae and semen Abutili, extracting by supercritical carbon dioxide extraction, and pulverizing to obtain extract I powder;
percolating the above residue, drying, and pulverizing to obtain extract II powder;
mixing the extract I powder and the extract II powder uniformly to obtain patch ointment;
mixing oleum Menthae Dementholatum, camphora, vanillyl butyl ether, light liquid paraffin and methyl salicylate to obtain wetting solvent.
The invention mixes raw kusnezoff monkshood root, acorus calamus, costustoot, liquidambar formosana hance, cassia seed and abutilon seed, and then carries out supercritical carbon dioxide extraction and crushing to obtain extract I medicinal powder.
Preferably, after mixing the raw kusnezoff monkshood root, the acorus calamus, the costustoot, the liquidambar formosana hance, the cassia seed and the abutilon seed, the method further comprises the following steps: pulverizing, and sieving with 60 mesh sieve.
Preferably, the temperature of the supercritical extraction is 25-35 ℃, the pressure is not lower than 7.4MPa, and the time is 2-3 h. Specifically, the supercritical extraction temperature is 30 ℃, the pressure is 8.0MPa, and the time is 2 hours or 3 hours.
Preferably, the method further comprises the steps of: sieving with 100 mesh sieve.
In some embodiments of the present invention, after mixing radix Aconiti Kusnezoffii, rhizoma Acori Calami, radix aucklandiae, resina Liquidambaris, semen Cassiae and semen Abutili, performing supercritical carbon dioxide extraction, and pulverizing to obtain extract I powder, which specifically comprises:
mixing radix Aconiti Kusnezoffii, rhizoma Acori Calami, radix aucklandiae, resina Liquidambaris, semen Cassiae and semen Abutili, pulverizing, sieving with 60 mesh sieve, adding into supercritical extraction tank, pressing carbon dioxide liquid, performing supercritical extraction, pressing carbon dioxide-containing liquid back into volatilization tank, relieving pressure, volatilizing carbon dioxide to obtain solid, pulverizing, and sieving with 100 mesh sieve to obtain extract I powder.
The medicine dregs after supercritical extraction are subjected to diacolation extraction, dried and crushed to obtain extract II medicine powder.
In certain embodiments of the invention, the agent employed in the percolation extraction is an ethanol solution. Preferably, the mass concentration of the ethanol solution is 75%.
In certain embodiments of the invention, the percolation rate of the percolation extraction is 20-30 mL/kg; specifically, the concentration may be 20mL/kg or 30mL/kg.
Preferably, the percolation extraction includes:
placing the residue after supercritical extraction into a percolating tank, adding ethanol solution, soaking, percolating at a percolating speed of 20-30 mL/kg, concentrating under reduced pressure, and recovering ethanol to obtain fluid extract.
Preferably, the soaking time is 2-4 hours; specifically, the time may be 3 hours.
Preferably, the percolation time is 22-26 hours; specifically, the time may be 24 hours.
In some embodiments of the present invention, the ethanol solution may be replenished during the percolation process so that the ethanol solution is free of residues.
In certain embodiments of the invention, the fluid extract has a relative density of 1.18 to 1.21 at 60 ℃.
And (3) after obtaining the fluid extract, drying and crushing to obtain extract II medicinal powder.
In certain embodiments of the invention, the drying is freeze drying. Specifically, the fluid extract is spread into a drying disc with the thickness of 1-2 cm, and the fluid extract is put into a freeze dryer for freeze drying.
In certain embodiments of the invention, the extract II powder is obtained by pulverizing and sieving with a 100 mesh sieve.
And after the extract I powder and the extract II powder are obtained, uniformly mixing the extract I powder and the extract II powder to obtain the patch ointment.
In certain embodiments of the invention, the blending is performed in a three-dimensional motion mixer. The mixing time is 15-25 min, specifically, 20min.
Preferably, after the extract I powder and the extract II powder are uniformly mixed, the method further comprises the following steps:
and subpackaging the obtained mixed medicinal powder into non-woven fabric bags, attaching one surface of the non-woven fabric bags to the center of a non-woven fabric adhesive tape, attaching a simethicone polyester film, and filling into an aluminum-plastic composite bag to obtain the patch ointment storage bag.
In certain embodiments of the invention, the nonwoven bag is a square or rectangular nonwoven bag and the nonwoven tape is a rectangular nonwoven tape. The split charging amount of the mixed medicinal powder in the non-woven bag is 1.0 g/bag.
The invention evenly mixes peppermint oil, camphor, vanillyl butyl ether, light liquid paraffin and methyl salicylate to obtain the wetting solvent.
Preferably, the wetting solvent is prepared according to the following method:
adding oleum Menthae Dementholatum, camphora, vanillyl butyl ether and light liquid paraffin into methyl salicylate, stirring, dissolving, clarifying, and packaging to obtain wetting solvent.
In some embodiments of the invention, the packaging is carried out in aluminum plastic composite bags, and the packaging amount is 2 mL/bag.
The invention also provides a using method of the emplastrum, which comprises the following steps of:
spraying the wetting solvent on the patch ointment bag, and applying the obtained patch to the affected part after absorption.
Specifically, the method comprises the following steps:
taking a patch ointment storage bag, removing the package, tearing off the simethicone polyester film, opening the aluminum-plastic composite bag filled with the wetting solvent, spraying the wetting solvent on a non-woven fabric bag filled with the patch ointment, and sticking the obtained patch on an affected part after the wetting solvent is absorbed by the patch ointment, and compacting.
The raw material components used in the present invention are not particularly limited, and may be generally commercially available.
The emplastrum provided by the invention has no skin irritation, basically no sensitization reaction, good safety and better anti-inflammatory and analgesic effects, can be used for treating rheumatic arthritis, pain in the abdomen, soreness of waist and legs and migraine, and has better curative effect.
Detailed Description
The technical solutions of the present invention will be clearly and completely described in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
The raw material formula comprises:
the raw material formula of the patch ointment comprises the following components: 75kg of raw kusnezoff monkshood root, 52.5kg of acorus calamus, 52.5kg of costustoot, 32.5kg of sweetgum fruit lipid, 52.5kg of cassia seed and 52.5kg of abutilon seed;
the raw material formula of the wetting solvent comprises: 11.9kg of peppermint oil, 19.7kg of camphor, 10kg of vanillyl butyl ether, 10kg of methyl salicylate and 27kg of light liquid paraffin;
the preparation method comprises the following steps:
(1) Mixing radix Aconiti Kusnezoffii, rhizoma Acori Calami, radix aucklandiae, resina Liquidambaris, semen Cassiae and semen Abutili, pulverizing, sieving with 60 mesh sieve, adding into supercritical extraction tank, pressing into carbon dioxide liquid, controlling temperature at 30deg.C and pressure at 8.0MPa, supercritical extracting for 2 hr, pressing carbon dioxide-containing liquid into volatilizing tank, pressure releasing, volatilizing carbon dioxide to obtain solid, pulverizing, sieving with 100 mesh sieve to obtain extract I powder;
(2) Placing the residue after supercritical extraction into a percolating tank, adding 75% ethanol to the residue, soaking for 3 hr, percolating at 20mL/kg for 24 hr, supplementing 75% ethanol solution during the percolating process, concentrating under reduced pressure, and recovering ethanol to obtain fluid extract (relative density of 1.19 at 60deg.C); spreading the fluid extract into a drying disc with the thickness of 1-2 cm, putting into a freeze dryer for pre-freezing, freeze-drying, crushing and sieving with a 100-mesh sieve to obtain extract II medicinal powder;
(3) Adding the extract I powder and the extract II powder into a three-dimensional motion mixer, mixing for 20min, subpackaging the obtained mixed powder into square non-woven fabric bags (1.0 g/bag), attaching one side of the non-woven fabric bags to the center of a rectangular non-woven fabric adhesive tape, attaching a simethicone polyester film, and filling into an aluminum-plastic composite bag to obtain patch ointment storage bags;
(4) Adding methyl salicylate into a preparation tank, adding peppermint oil, camphor, vanillyl butyl ether and light liquid paraffin into the stirred methyl salicylate, stirring, dissolving, clarifying, and packaging into aluminum-plastic composite bags (2 mL/bag) to obtain a wetting solvent.
Example 2
The difference from example 1 is that: the supercritical extraction time is 3h.
Example 3
The difference from example 2 is that: the percolation speed of percolation extraction is 30mL/kg.
Comparative example 1
The raw material formula comprises:
the raw material formula of the patch ointment comprises the following components: 75kg of raw kusnezoff monkshood root, 52.5kg of acorus calamus, 52.5kg of costustoot, 32.5kg of sweetgum fruit lipid, 52.5kg of cassia seed and 52.5kg of abutilon seed;
the auxiliary materials comprise the following formula: 11.9kg of peppermint oil, 19.7kg of camphor, 133kg of rubber, 133kg of rosin and 27kg of light liquid paraffin;
the preparation method comprises the following steps:
pulverizing the raw materials of the patch ointment into coarse powder, soaking in 95% ethanol for 24 hr, filtering, mixing filtrates, and concentrating into thick paste;
adding oleum Menthae Dementholatum, camphora and the thick paste into matrix prepared by heating and melting rubber, colophonium and light liquid paraffin, kneading into paste by kneader, and making into paste;
coating the paste on a simethicone-coated polyester film, sealing the four-sided elastic non-woven fabric on the coated polyester film, cutting into square pieces with the thickness of 70cm multiplied by 10cm, and obtaining the plaster with the paste content of not less than 1.6 g.
Comparative example 2
The difference from comparative example 1 is that:
the raw material formulation of the patch ointment comprises:
75kg of raw kusnezoff monkshood root, 52.5kg of acorus calamus, 52.5kg of costustoot, 32.5kg of sweetgum fruit lipid and 52.5kg of abutilon seed.
Comparative example 3
The difference from example 1 is that:
the preparation method comprises the following steps:
(1) Mixing radix Aconiti Kusnezoffii, rhizoma Acori Calami, radix aucklandiae, resina Liquidambaris, semen Cassiae and semen Abutili, pulverizing, sieving with 60 mesh sieve, adding 75% ethanol, soaking for 3 hr, percolating at 20mL/kg for 24 hr, concentrating under reduced pressure, recovering ethanol to obtain fluid extract (relative density of 1.19 at 60deg.C); spreading the fluid extract into a drying disc with the thickness of 1-2 cm, freeze-drying in a freeze dryer, crushing and sieving with a 100-mesh sieve to obtain extract I powder;
(2) Adding the extract I powder into a three-dimensional motion mixer, mixing for 20min, subpackaging the obtained mixed powder into square non-woven fabric bags (1.0 g/bag), attaching one side of the non-woven fabric bags to the center of a rectangular non-woven fabric adhesive tape, attaching a simethicone polyester film, and filling into an aluminum-plastic composite bag to obtain patch ointment storage bags;
(3) Adding methyl salicylate into a preparation tank, adding peppermint oil, camphor, vanillyl butyl ether and light liquid paraffin into the stirred methyl salicylate, stirring, dissolving, clarifying, and packaging into aluminum-plastic composite bags (2 mL/bag) to obtain a wetting solvent.
Comparative example 4
The difference from comparative example 3 is that:
the raw material formulation of the patch ointment comprises:
75kg of raw kusnezoff monkshood root, 52.5kg of acorus calamus, 52.5kg of costustoot, 32.5kg of sweetgum fruit lipid and 52.5kg of abutilon seed.
The safety and beneficial effects of the invention are demonstrated by experimental examples below.
Test example 1
1 skin irritation test
The allergic stimulus test was performed, and specific detection methods and results are as follows:
test drug: the patches prepared in examples 1 to 3;
the method of using the patch described in embodiments 1 to 3 includes:
taking a patch ointment storage bag, removing the package, tearing off the simethicone polyester film, opening the aluminum-plastic composite bag filled with the wetting solvent, spraying the wetting solvent on a non-woven fabric bag filled with the patch ointment, and sticking the obtained patch on an affected part after the wetting solvent is absorbed by the patch ointment, and compacting.
Experimental animals: white guinea pigs, male and female halves, body weight (250.+ -.20) g.
1. Test method (Experimental example 1 below used the patch prepared in example 1, experimental example 2 used the patch prepared in example 2, and experimental example 3 used the patch prepared in example 3)
(1) Acute skin toxicity test
Healthy white guinea pigs (weight of 250+/-20 g) are taken, and male and female guinea pigs are randomly divided into 2 large groups (namely a complete skin group and a broken skin group) according to the sex weight sequence, wherein each large group is divided into 7 small groups, namely a complete skin group (a physiological saline group 0.2 g/group, a test example 1 low dose group, a test example 1 high dose group, a test example 2 low dose group, a test example 2 high dose group, a test example 3 low dose group and a test example 3 high dose group, wherein the low high dose is 0.2 g/group and 0.5 g/group respectively, and the ointment is ointment with uniform wetting of medicinal powder and a wetting agent), and a broken skin group (physiological saline 0.5 g/group, a test example 1 low dose group, a test example 1 high dose group, a test example 2 low dose group, a test example 2 high dose group, a test example 3 low dose group and a test example 3 high dose group are respectively, and the low high dose is 0.2 g/group and 0.5 g/group respectively), and each group is 10. After 24 hours before administration, the back of guinea pigs is dehaired conventionally, the dehairing range is about 10cm multiplied by 4cm, and after dehairing for 24 hours, animals in each group are respectively smeared with ointment with different concentrations for contamination. The damaged skin group guinea pigs are scratched on the dehaired parts by using fine sand paper until the skin has a slightly dense bleeding point. Within 24 hours, the number of animal application times was 1. After the infection, the back of the guinea pig is coated with a breathable film and gauze, and is wrapped and fixed by adhesive tape, and the guinea pig is fed in separate cages. After 24 hours, the residual test agent was removed with warm water. After removal of the residual test substance, the animal was continuously observed for 1 week to record changes in body weight, skin hair, eyes and mucous membrane, respiration, central nervous system, limb activities, etc. and other toxic manifestations.
(2) Acute skin irritation test
Healthy white guinea pigs (250+/-20) g, male and female half, are divided into two large groups of a complete skin experimental group and a damaged skin experimental group, and the two large groups are respectively divided into 4 small groups (physiological saline group, experimental example 1 group, experimental example 2 group and experimental example 3 group). Hairing is carried out on two sides of the back of a guinea pig, and the hairing area of one side is 4cm multiplied by 5cm. The method comprises the steps of adopting a self-comparison method on the left side and the right side of a body, coating a test object on the skin of a guinea pig at a normal dose, coating an equivalent amount of corresponding blank matrix on the skin of the guinea pig at the right side of the body as a control, covering the skin with gauze, fixing the skin with adhesive tape, cleaning the residual medicine with warm water after the skin is continuously used for 24 hours, observing the skin of the test object for 1 hour, 24 hours, 48 hours and 72 hours after the medicine is cleaned, and recording whether red spots, edema and the like exist at the coating position. The animals were scored according to the skin irritation response scoring criteria to calculate the average score for each group of animals. If damage and other toxic reactions are to be recorded in detail, see tables 1 and 2.
Average score of response = (total score of erythema formation ± total score of edema formation)/animal number.
TABLE 1 irritation symptom scoring criteria
TABLE 2 skin irritation intensity evaluation criteria
| Average score | Evaluation |
| 0-0.49 | Without stimulation |
| 0.5-2.99 | Mild irritation |
| 3.0-5.99 | Moderate irritation |
| 6.0-8.0 | Strong irritation |
(3) Acute skin allergy test
Healthy white guinea pigs were divided into a whole skin group (normal saline group, experimental example 1 group, experimental example 2 group, experimental example 3 group, positive drug control group), and a damaged skin group (normal saline group, experimental example 1 group, experimental example 2 group, experimental example 3 group, positive drug control group), each group of 10. The hairing agent was used to remove hairs on both sides of the back spine of the guinea pig 24 hours before administration, and the area of hair removal on one side was 3cm×3cm.
(1) Sensitization contact experiments 1 to 3 were carried out by applying 0.2g of an adhesive plaster (the plaster was a plaster having a uniform wetting of a powder and a wetting agent), 0.2g of a physiological saline group, and 0.2ml of 1%2, 4-dinitrochlorobenzene to a positive control group to the left dehairing area, covering with gauze and fixing with an adhesive tape, washing the residual medicine with warm water after lasting for 6 hours, and rearing in separate cages, and repeating the same manner for 3 times on days 7 and 14.
(2) On day 14 after sensitization of the last dose, 0.2g of the test sample, 0.2g of the corresponding physiological saline group and 0.2mL of the positive drug (2, 4-dinitrochlorobenzene solution with the excitation concentration of 0.1%) were used for the right-side test skin corresponding to the 3 groups of guinea pigs in the same way as the method (1), the drug was cleared and observed immediately after 6 hours of drug administration, and then the skin allergic reaction condition was observed again 24 hours, 48 hours and 72 hours after drug removal. Each group was scored according to the scoring criteria for skin allergy degree. Meanwhile, attention is paid to observing whether guinea pigs have systemic allergic reactions such as asthma, unstable standing and the like, see tables 3 and 4.
Average score of response = (total score of erythema formation ± total score of edema formation)/number of animals,
sensitization incidence = number of animals with allergic reaction/total number of animals x 100%.
TABLE 3 skin allergy scoring criteria
TABLE 4 evaluation criteria for skin sensitization
| Incidence of sensitization (%) | Evaluation |
| 0-10 | No sensitization |
| 11-30 | Mild sensitization |
| 31-60 | Moderate sensitization |
| 61-80 | Highly sensitized |
| 81-100 | Extremely high sensitization |
2. Test results
(1) Acute skin toxicity test results
The results of the study show that the indexes such as fur, eyes, respiration, diet, stool and urine, autonomous activities and the like of each group of guinea pigs show that no abnormal condition is found between the complete skin group and the damaged skin group and between the low-dose group and the high-dose group of the reagent and the comparison with the normal saline group, no obvious difference is found between the complete skin group and the damaged skin group, and no poisoning and death condition are found. The clear difference in weight gain between groups compared to the corresponding matrix groups was statistically significant (P > 0.05) for the groups, and no acute skin toxicity was observed in the transdermal examples and comparative examples at the above doses, indicating that the patch was very safe for use with the Mongolian drug, see Table 5.
TABLE 5 Effect of acute toxicity on guinea pig body weight
(2) Acute skin irritation test results
The results show that after the experimental example group and the normal saline group continuously act on the intact skin and the damaged skin of guinea pigs for 6 hours, barely visible erythema or edema can appear in the experimental example group, the experimental example group can quickly disappear by oneself, other animals have no erythema, edema and other conditions, and the average response values of the skin irritation degree of the groups are as follows: the whole skin group, experimental examples 1 to 3 were 0.125, 0.125 and 0.25, respectively; experimental examples 1 to 3 of the broken skin groups were 0.125, 0.25 and 0.25, respectively, less than 0.49, and no irritation was observed. The results show that the products of examples 1-3 are free of skin irritation, and in particular that the average response to the degree of irritation of example 1 is the lowest. The results are shown in Table 6.
TABLE 6 local skin irritation response to guinea pigs
(3) Test results of sensitization to skin
The physiological saline group does not have skin allergic conditions such as erythema, edema and the like; experimental examples few animals had mild erythema and edema reactions after drug removal, and the sensitization rates were as follows: experimental examples 1-3 show that the sensitization rate of intact skin and damaged skin is 10% at the highest, and the positive control group shows obvious skin allergic conditions such as erythema, edema and the like, and the integral value is high, and the sensitization rate reaches 100%. The results show that the experimental examples have substantially no sensitization reaction to the skin, and the results are shown in table 7.
TABLE 7 Effect on skin sensitization Rate in guinea pigs
Test example 2 Primary pharmacodynamic experiments for pain relief and inflammation diminishing
The anti-inflammatory and analgesic effects of the plasters prepared in each comparative example and example were examined, and specific examination methods and results are shown below:
1. test method
(1) Rat plantar egg white swelling experiment
Experimental drugs: each comparative example and example drug, physiological saline; the positive drug was indomethacin Xin Chaji (1.5%).
Experimental animals: SD rats (200+ -10 g), male and female halves.
100 SD rats were randomly divided into 10 groups of 10 animals (female half), and the groups were blank group, normal saline group, example 1, example 2, example 3, comparative example 1, comparative example 2, comparative example 3, comparative example 4, and positive drug indomethacin group. Saline group, example group, comparative example group each rat right hind paw was given 2.5cm 2 0.1mL of physiological saline or 0.1g of ointment, 0.1mL of indomethacin Xin Chaji (1.5%) applied to right hind foot of a positive drug group rat, and the application area is 2.5cm 2 . The administration was performed 1 time a day for 4 hours, and after the completion of each administration, the test agent was removed and the administration site was cleaned with warm water, followed by continuous administration for 3 days. After completion of the last administration for 1h, 100% fresh egg white was subcutaneously injected in the right hind leg to cause inflammation in 0.1 mL. And after molding, measuring the circumference of the 0.5cm position below the ankle joint of the right hind limb by using cotton threads and a graduated scale at the 0.5h, 1h, 2h, 4h and 6h, and calculating the swelling rate. Swelling ratio (%) = (circumference measured at a certain point after molding-circumference before molding)/circumference before molding×100%.
(2) Mouse auricle xylene inflammation experiment
Experimental drugs: and (3) performing a swelling experiment with the egg white of the foot of the rat.
Experimental animals: kunming mice (22.+ -.2 g), male and female halves.
100 Kunming mice were taken, each half of the male and female, and randomly divided into 10 groups of 10 mice (each half of the female), and the groups were a blank group, normal saline, example 1, example 2, example 3, comparative example 1, comparative example 2, comparative example 3, comparative example 4, and indomethacin positive drug, respectively. Each of the groups of examples, comparative examples and physiological saline groups was coated with 0.1mL of matrix or ointment before and after the auricles of each group of rats, and the positive group of rats was coated with 0.1mL of indomethacin Xin Chaji (1.5%) for 4 hours, and after the end of the administration, the test drug was removed and the administration site was cleaned with warm water. After the administration was completed for 1 hour, 0.02mL of xylene was applied to both front and rear sides of the right ear of each mouse, the left ear was used as a control, the animals were euthanized after 30 minutes, the two ears were cut off, the same parts of the two ears were respectively punched out with a punch having a diameter of 5mm, and the animals were weighed. The swelling degree is obtained by subtracting the mass of the left ear from the mass of the right ear of each mouse.
(3) Pain relieving experiment by hot plate method
Experimental drugs: and (3) performing a swelling experiment with the egg white of the foot of the rat.
Experimental animals: kunming mice (22.+ -.2 g), male and female halves.
The Kunming mice were placed on a preheated stainless steel hotplate (55.+ -. 0.5 ℃ C.) and mice with a basal pain threshold of 5-30 s were selected for the experiment. The mice were grouped with auricle xylene inflammatory experiments, and the basal pain threshold was measured again the next day as the pre-dose pain threshold. The distance of 1cm between the two hind feet of the mice is the administration part, 0.1mL of drug or physiological saline or positive drug is smeared, and after the administration is finished, the tested drug is removed and the administration part is cleaned by warm water. The pain threshold was measured 1 time after 10min and 60min after completion of dosing, respectively, and compared to the basal pain threshold before dosing.
(4) Experiment of Effect on mouse auricle Small vessels
Experimental drugs: and (3) performing a swelling experiment with the egg white of the foot of the rat.
Experimental animals: and (5) carrying out analgesic experiments by a hot plate method.
Mice were grouped with rat plantar egg white swelling experiments. Each group of rats in the examples, the comparative examples and the normal saline groups was coated with 0.2g of normal saline or ointment before and after auricles of both ears, and the positive drug group was coated with 0.2mL of indomethacin Xin Chaji (1.5%). Abdominal injection of pentobarbital sodium (0.5 mg/10 g) into mice, sticking auricle and villus to the mice with medical adhesive tape after anesthesia, and making the ambient temperature of the mice about 36-38 ℃ with a large incandescent lamp. After 10min, removing the test medicine, cleaning the administration part with warm water, dripping a little liquid paraffin, immediately placing under a microscope, and observing and recording the diameters of auricle artery and vein blood vessel with a 40-time microscope and a micrometer.
2. Test results
(1) Experimental results of rat plantar egg white induced swelling
The pharmacodynamic effects of each example group and comparative example group were observed by injecting egg white into the plantar region of the rat to induce swelling, and the results are shown in table 8. The example groups were all able to significantly reduce the swelling rate of the plantar region. Both the examples 1-3 and the positive drug group significantly reduced the swelling rate of the plantar region (P < 0.001) compared to the blank control group; the comparative examples 1 and 3 significantly reduced the swelling rate of the plantar region (P < 0.01) compared to the blank, and the comparative examples 2 and 4 significantly reduced the swelling rate of the plantar region (P < 0.05). In summary, the groups of examples 1 to 3 have obvious effect of reducing the swelling caused by the plantar egg white of the rat, and the experimental group has better effect than the comparative group.
Table 8 changes in the rate of plantar swelling (%)
In table 8, P <0.05, P <0.01, P <0.001 compared to the blank.
(2) Experimental results of auricle xylene inflammation
The pharmacodynamic effects of each of the example groups and the comparative example group were observed by applying xylene inflammation to the auricle of the mice, and the results are shown in Table 9. Experimental results show that each group of drugs can resist swelling of the ear caused by xylene, but the therapeutic effects are different, specifically as follows: examples 1 to 3 (P < 0.01), comparative example 1 and comparative examples 3 to 4 (P < 0.05), positive drug group (P < 0.01). The examples are shown to have a pronounced anti-inflammatory effect, and the experimental examples are more effective than the comparative examples.
TABLE 9 Effect of paraxylene-induced mouse auricle inflammation
P <0.05, < P <0.01 compared to the placebo group.
(3) Pain threshold detection result by hot plate method
Analgesic efficacy was measured by thermal stimulation for each of the example groups and the comparative example group, and the results are shown in table 10. The pain threshold (P < 0.01) of rats to thermal stimulation was significantly increased in groups 1-3, both 10min and 60min post-dose; the groups of comparative examples 1 to 4 had no significant effect on the pain threshold of the rats to thermal stimulation 10min after the administration, and the groups of comparative examples 1 and 3 significantly reduced the pain threshold of the rats to thermal stimulation 60min after the administration (P < 0.01), and the group of comparative example 2 (P < 0.05). In summary, the examples have a significant analgesic effect, the comparative examples have a certain analgesic effect, and the group of examples has a better analgesic effect than the group of comparative examples.
Table 10 analgesic effect on mice (hotplate method)
P <0.05, P <0.01 compared to prior to dosing in this group.
(4) Results of drug effects on auricular small vessels
The effect of the drug on local microcirculation was judged by observing the change in small blood vessel diameter of auricle of the mice after administration, and the results are shown in Table 11. Examples 1-3 and comparative example 3 were able to significantly dilate local arterioles and venules (P < 0.05), whereas no significant change in vessel diameter was observed in comparative example 1, 2,4 and the positive drug indomethacin group.
TABLE 11 auricle small vessel diameter of mice of each group
P <0.05 compared to the placebo group.
Test example 3 pharmacodynamic test for migraine treatment
Materials: SPF-class SD male rats, 48, body weight (200+ -20) g, were supplied by Chongqing medical university animal house. The experimental animals were kept in a barrier environment at a temperature of 25℃and a humidity of 50%.
Experimental samples: physiological saline, sumatriptan, nitroglycerin injection, example 2 sample
The test method comprises the following steps: male SD rats were randomly divided into a blank control group, a model control group (model of migraine induced by nitroglycerin), a positive drug group [ sumatriptan succinate 5.8 mg/(kg.d) ], a low dose group of example 2 [0.1 g/(kg.d) ] and 1g of medicinal powder was taken and 1mL of wetting agent was added for uniform wetting for the test ], a dose group of example 2 [0.2 g/(kg.d) ], a high dose group of example 2 [0.4 g/(kg.d) ], and 6 animals per group. After adaptively raising rats before experiments, fixing the rats, shaving the heads, applying the medicine on the heads, fixing the medicine by a non-woven fabric adhesive tape, and continuously dosing for 5d and 1 time/d according to doses. The blank and model groups were perfused with physiological saline, the positive drug control group was perfused with sumatriptan, the dosing group was coated with the example 2 wet sample, and fixed with a non-woven tape. 30min after the 5d administration, 10mg/kg of nitroglycerin injection was subcutaneously injected in the frontal area of the rats. 2h after molding, 3mL/kg of 10% chloral hydrate is injected into the abdominal cavity for anesthesia, and the abdominal aorta is taken for blood, and serum is separated. Cervical vertebrae are removed after blood is taken out, brainstem is separated, and the brainstem is frozen.
5-hydroxytryptamine (5-HT), calcitonin Gene Related Peptide (CGRP), beta-endorphin (beta-EP), endothelin-1 (ET-1), matrix metalloproteinase-9 (MMP-9), nuclear factor κB (NF- κB), nitric Oxide Synthase (NOS) content assay: the enzyme-linked immunosorbent assay (ELISA) is used for measuring the contents of 5-HT, CGRP, beta-EP, ET-1, MMP-9 and NF- κB of left brainstem and measuring the activity of NOS in serum.
Relative expression analysis of Calca and c-fos mRNA Trizol method was performed to extract total RNA from the right brainstem. 1 mug total RNA template is prepared into 20 mug reaction system to synthesize first chain cDNA according to the instruction of quick reverse transcription reagent kit, and GAPDH gene is used as reference gene. The reaction conditions were 42℃for 15min,95℃for 3min. Taking 0.8 mu L of cDNA template, 0.6 mu L of each of the upstream primer and the downstream primer, preparing 20 mu L of reaction system according to the instruction of an amplification kit, and amplifying, wherein the PCR thermal cycle parameters are 1) 1X of pre-denaturation at 95 ℃ for 15min; 2) PCR at 40X,95 ℃,10s,52 ℃,20s,72 ℃ for 30s; 3) And (3) carrying out melting curve analysis, namely arranging 3 compound holes in parallel for each sample, and carrying out relative expression analysis on the Calca and c-fos mRNA of each sample.
The results of HT, beta-EP, CGRP, ET-1, MMP-9, NF-kappa B, NOS assays are shown in Table 12. Compared with the blank control group, the 5-HT and beta-EP contents of the rats in the model control group are obviously reduced (P is less than 0.01 and P is less than 0.01), and the contents are obviously increased (P is less than 0.05, P is less than 0.01 and P is less than 0.01). Compared with the model control group, the contents of 5-HT and beta-EP of the positive medicine groups are obviously increased (P is less than 0.01 ), and the contents of CGRP, ET-1, MMP-9 and NF-B, NOS are obviously reduced (P is less than 0.05, P is less than 0.01, and P is less than 0.01). Compared with a model control group, the XFDP low-dose group can obviously raise the contents of 5-HT and beta-EP (P is less than 0.01 and P is less than 0.01), obviously reduce the contents of CGRP, MMP-9 and NF-kappa B, NOS (P is less than 0.05, P is less than 0.01 and P is less than 0.01), and can reduce the content of ET-1 but has no statistical significance. Compared with the model control group, the ointment of the example 2 can obviously raise the contents of 5-HT and beta-EP (P is less than 0.01 and P is less than 0.01), and obviously reduce the contents of CGRP, ET-1, MMP-9 and NF-kappa B, NOS (P is less than 0.01, P is less than 0.05, P is less than 0.01 and P is less than 0.01). The results demonstrate that the ointment of example 2 can exert an anti-migraine effect by increasing the 5-HT and beta-EP content in brainstem, decreasing the CGRP, ET-1, MMP-9 and NF- κB content in brainstem, and decreasing the NOS content in serum.
TABLE 12 migraine rats 5-HT, CGRP, beta-EP, ET-1, MMP-9, NF- κ B. NOS content
Comparing P < 0.05.p <0.01 with the model control group, P <0.05.0P <0.01
The results of the Calca and c-fos mRNA measurements are shown in Table 13. Compared with the blank control group, the expression of the Calca mRNA and the c-fos mRNA of the model control group rats is obviously increased (P is less than 0.05 and P is less than 0.1). Compared with the model control group, the expression of the mRNA of the Calca and the c-fos of the positive drug group is obviously reduced (P is less than 0.05 and P is less than 0.05). Compared with the model control group, the low, medium and high dose groups of the medicine of the example 2 can obviously reduce the expression of the mRNA of Calca and c-fos (P is less than 0.05 and P is less than 0.05). The results demonstrate that the drug of example 2 can exert an anti-migraine effect by reducing the expression of Calca, c-fos mRNA in the brainstem.
TABLE 13 expression of migraine rat brainstem Calca and c-fos mRNA
| Group of | Calca | c-fos |
| Model control group | 1.68±0.20 * | 1.77±0.16** |
| Blank control group | 0.56±0.06 Δ | 1.01±0.06 ΔΔ |
| Positive medicine group | 0.79±0.15 Δ | 1.25±0.09 Δ |
| Low dose group | 0.81±0.05 Δ | 1.39±0.10* Δ |
| Medium dose group | 0.73±0.22 Δ | 1.28±0.06 Δ |
| High dose group | 0.65±0.13 Δ | 1.04±0.05 ΔΔ |
Note that P <005, < P <0.01 compared to the placebo group; Δp <0.05, ΔΔp <0.01 compared to model control.
Experimental results show that the medicine in the embodiment 2 can increase the 5-HT and beta-EP contents of migraine rats, reduce the contents of CGRP, ET-1, MMP-9 and NF-kappa B, NOS, reduce the expression of the mRNA of Calca and c-fos, and play a role in treating migraine.
Example 2 a pharmaceutical product exerts a therapeutic effect on migraine by modulating the expression of various neurotransmitters and vasoactive substances.
The above description of the embodiments is only for aiding in the understanding of the method of the present invention and its core ideas. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (9)
1. A Mongolian medicine emplastrum is suitable for migraine, and comprises a emplastrum ointment and a wetting solvent;
the patch ointment is prepared from the following raw materials in parts by weight: 60-80 parts of raw kusnezoff monkshood root, 50-60 parts of rhizoma acori graminei, 50-60 parts of elecampane, 30-40 parts of liquidambar formosana hance, 50-60 parts of semen cassiae and 50-60 parts of abutilon seed;
the wetting solvent consists of, by weight, 10-20 parts of peppermint oil, 15-25 parts of camphor, 10-20 parts of vanillyl butyl ether, 10-20 parts of methyl salicylate and 20-30 parts of light liquid paraffin.
2. The patch according to claim 1, wherein the patch is prepared from the following raw materials in parts by weight:
75 parts of raw kusnezoff monkshood root, 52.5 parts of rhizoma acori graminei, 52.5 parts of costustoot, 32.5 parts of liquidambar formosana hance, 52.5 parts of cassia seed and 52.5 parts of abutilon seed;
the wetting solvent consists of 11.9 parts of peppermint oil, 19.7 parts of camphor, 10 parts of vanillyl butyl ether, 10 parts of methyl salicylate and 27 parts of light liquid paraffin according to parts by weight.
3. The method for preparing the patch according to claim 1 or 2, comprising the steps of:
mixing radix Aconiti Kusnezoffii, rhizoma Acori Calami, radix aucklandiae, resina Liquidambaris, semen Cassiae and semen Abutili, extracting by supercritical carbon dioxide extraction, pressing carbon dioxide-containing liquid into volatilizing pot, relieving pressure, volatilizing carbon dioxide to obtain solid, and pulverizing to obtain extract I powder;
percolating the above residue, drying, and pulverizing to obtain extract II powder;
mixing the extract I powder and the extract II powder uniformly to obtain patch ointment;
mixing oleum Menthae Dementholatum, camphora, vanillyl butyl ether, light liquid paraffin and methyl salicylate to obtain wetting solvent.
4. The preparation method according to claim 3, wherein after mixing the raw kusnezoff monkshood root, the acorus calamus, the costustoot, the liquidambar formosana hance, the cassia seed and the abutilon seed, the preparation method further comprises: pulverizing, and sieving with 60 mesh sieve.
5. The preparation method according to claim 3, wherein the supercritical extraction is performed at a temperature of 25-35 ℃ and a pressure of not less than 7.4MPa for 2-3 hours;
through supercritical carbon dioxide extraction, will contain carbon dioxide liquid and press back the jar that volatilizees, the pressure release lets carbon dioxide volatilize, obtains the solid matter, after smashing, still includes: sieving with 100 mesh sieve.
6. A method of preparing according to claim 3, wherein the percolation extraction comprises:
and (3) filling the medicinal residues after supercritical extraction into a percolating tank, adding ethanol, soaking, percolating at a percolating speed of 20-30 mL/kg, concentrating under reduced pressure, and recovering ethanol to obtain fluid extract.
7. The method according to claim 6, wherein the mass concentration of ethanol is 75%;
the soaking time is 2-4 hours;
the percolating time is 22-26 hours.
8. The preparation method according to claim 3, wherein after the extract i powder and the extract ii powder are mixed uniformly, the preparation method further comprises:
and subpackaging the obtained mixed medicinal powder into non-woven fabric bags, attaching one surface of the non-woven fabric bags to the center of a non-woven fabric adhesive tape, attaching a simethicone polyester film, and filling into an aluminum-plastic composite bag to obtain the patch ointment storage bag.
9. A method according to claim 3, wherein the wetting solvent is prepared according to the following method:
adding oleum Menthae Dementholatum, camphora, vanillyl butyl ether and light liquid paraffin into methyl salicylate, stirring, dissolving, clarifying, and packaging to obtain wetting solvent.
Priority Applications (1)
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| CN202211285936.4A CN115463186B (en) | 2022-10-20 | 2022-10-20 | Mongolian medicine emplastrum, preparation method and application thereof |
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| CN202211285936.4A CN115463186B (en) | 2022-10-20 | 2022-10-20 | Mongolian medicine emplastrum, preparation method and application thereof |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105477223A (en) * | 2015-12-28 | 2016-04-13 | 内蒙古科尔沁药业有限公司 | Mongolian medicinal ointment capable of relieving pain |
| CN111671871A (en) * | 2020-06-08 | 2020-09-18 | 娜仁 | Chinese herbal medicine for treating joint diseases by Mongolian donkey bone sibutrol therapy |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105477223A (en) * | 2015-12-28 | 2016-04-13 | 内蒙古科尔沁药业有限公司 | Mongolian medicinal ointment capable of relieving pain |
| CN111671871A (en) * | 2020-06-08 | 2020-09-18 | 娜仁 | Chinese herbal medicine for treating joint diseases by Mongolian donkey bone sibutrol therapy |
Non-Patent Citations (3)
| Title |
|---|
| 浅析蒙医药治疗类风湿关节炎临床方药特点;何心愉;中药药理与临床;第37卷(第6期);185-190 * |
| 热感剂的质量检测及肤感效果评价;陈少格;中国洗涤用品工业(第10期);第53页 * |
| 蒙药治疗偏头痛60例;刘萨仁;中国民族医药杂志;20011230(第04期);第10页 * |
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