CN112826851A

CN112826851A - Analgesic medicine and preparation method and application thereof

Info

Publication number: CN112826851A
Application number: CN202110172352.5A
Authority: CN
Inventors: 詹常森; 姜鹏; 董关涛; 温方方
Original assignee: Shanghai Hutchison Pharmaceuticals Co ltd
Current assignee: Shanghai Hutchison Pharmaceuticals Co ltd
Priority date: 2021-02-08
Filing date: 2021-02-08
Publication date: 2021-05-25

Abstract

The invention relates to the field of medical treatment, in particular to an analgesic drug and a preparation method and application thereof. The invention shows that the extract of the chopsticks has obvious effect on easing pain through deep experiments. The invention provides a new method and a new idea for analgesia.

Description

Analgesic medicine and preparation method and application thereof

Technical Field

The invention relates to the field of medical treatment, in particular to an analgesic drug, a preparation method and application thereof.

Background

Radix Chimonanthi Praecocis (Hellebori Radix Et Rhizoma) is dried root and rhizome of Radix Chimonanthi Praecocis (Helleborus thibetanus Franch.) of genus Radix Chimonanthi Praecocis of family Ranunculaceae (Ranunculaceae). It is recorded in Shaanxi province Standard of materia medica and Zhonghua Ben Uygur nationality medicated roll. The name is radix Papaveris, radix Caulophylli, Heihuaji, and Heihuaji. The Chinese medicinal herb is a special genuine medicinal material in Qinling region of China, is one of famous seven herbs in Shaanxi, is a perennial herb, has a stem height of 30-50 cm, and has no hair in the whole plant. The rootstock is short, and most dark brown fibrous roots grow under the rootstock. The stem is upright, the base part has membranous scales, and the upper part has few branches. 1-2 basal leaves with long handle, heart-shaped blade outline, 3 full clefts in the shape of bird feet, short handle, 3 full clefts in the center, inverted needle-shaped small clefts, and 2 full clefts on the lateral side; the cauline leaves have sheath-like short stems or almost no stems, and the leaves are smaller than the basal leaves and have 3 full clefts. Pollen is red, single, sometimes 2 plants live on the branch end; sepals 5, oval, lodged. The Gu whole-length fruit is flat, nearly oblong, 1.5-3cm long and has obvious transverse veins. Collected in early spring or autumn in flowering 4 months and fruit 5 months. Distributed in northwest Sichuan, southern Gansu, southern Shaanxi and northwest Hu provinces, and grown in mountain forest or bush with elevation of 1100-3700 m. The medicinal materials are slightly curled into a mass, the rootstocks are in a short cylindrical shape or branches are in a nodular shape, the length is 5-7 cm, the diameter is 3-7 mm, and the surface is dark brown or gray black. The lower part is coated with a plurality of dry, slightly flat and twisted fibrous roots, the diameter of which is 1-2 mm, and older ones are already cylindrical and are mostly brittle; hard, hard to break, yellow-white, woody, granular in cross section. Slight fishy smell, bitter taste, and numb tongue feeling after long-time chewing; has little toxicity.

The relation between the chopsticks and the analgesic effect does not appear in the prior art.

Disclosure of Invention

In view of the above-mentioned disadvantages of the prior art, the present invention aims to provide an analgesic, a preparation method and a use thereof, which provide a new idea for analgesia in the prior art.

The invention provides an application of an extract of Chimonanthus praecox in preparing an analgesic drug.

In another aspect of the invention, an analgesic drug is provided, wherein the active ingredient of the drug is an extract of Chimonanthus praecox.

The medicine necessarily comprises the pharmacodynamic components in the chopsticks. The pharmacodynamic components refer to components playing a therapeutic role.

In the medicine, the effective components playing the functions can be only the efficacy components of the chopsticks, and other molecules playing similar functions can also be contained.

The drug may be a multi-component substance.

The form of the drug is not particularly limited, and can be various forms of substances such as solid, liquid, gel, semifluid, aerosol and the like.

The drug is mainly aimed at mammals such as rodents, primates and the like.

Further, the chopsticks are prior art and commercially available to those skilled in the art.

Further, the extract of Chimonanthus praecox is an extract obtained by extracting Chimonanthus praecox with an extraction solvent.

The extraction solvent is not particularly limited as long as the effect of the present invention is not impaired, and may be water; monohydric alcohols such as methanol and ethanol; polyhydric alcohols such as 1, 3-butanediol and propylene glycol; lower alkyl esters such as ethyl acetate; ether, acetone, and the like. In the present invention, the above solvents may be used in combination. Preferred extraction solvents are water, methanol, ethanol, 1, 3-butanediol, or combinations thereof. Particularly preferred extraction solvents are water, ethanol, or a combination thereof (i.e., aqueous ethanol).

Further, the preparation method of the radix Chimonanthi Praecocis extract comprises the steps of adding 6-10 times of solvent into the medicinal materials, heating, refluxing, filtering, adding 4-8 times of solvent into filter residues, refluxing, extracting, filtering, combining the filtrates and concentrating. Further, the solvent is water or ethanol with the volume fraction of 30% -50%. Or 40% -50% of ethanol.

Further, the preparation method comprises adding 8 times of solvent into the medicinal materials, heating to boil, refluxing for 1h, filtering while hot, adding 6 times of solvent into the filter residue, refluxing for 1h, filtering, combining the filtrates and concentrating.

Furthermore, the concentration of the extract of the chopsticks in the medicine is 20-80 mug/ml, and can be 25 mug/ml, 30 mug/ml, 35 mug/ml, 40 mug/ml, 45 mug/ml, 50 mug/ml, 55 mug/ml, 60 mug/ml, 65 mug/ml, 70 mug/ml and 75 mug/ml.

In another aspect of the present invention, an analgesic drug combination is provided, comprising a therapeutically effective amount of an extract of Chimonanthus praecox and at least one other analgesic drug.

The combination therapy drug combination may be in any one of the following forms:

firstly), the extract of the radix Chimonanthi Praecocis and other analgesic drugs are respectively prepared into independent preparations, the preparation forms can be the same or different, and the administration routes can be the same or different.

When other analgesic drugs are chemical drugs, the administration forms can be rich, and the administration can be carried out in the gastrointestinal tract or the parenteral tract. Known routes of administration for each chemical are generally recommended.

And secondly) the radix Chimonanthi Praecocis extract and other analgesic drugs are prepared into a compound preparation, and when the radix Chimonanthi Praecocis extract and other analgesic drugs are administrated by the same administration route and are applied simultaneously, the radix Chimonanthi Praecocis extract and other analgesic drugs can be prepared into the compound preparation.

As described above, the analgesic drug of the present invention, the preparation method and the use thereof, have the following beneficial effects:

the invention discovers that the extraction of the chopsticks has the analgesic effect through experiments. The high dose Fr1 and the medium/high dose Fr3 can obviously improve the incubation period of light and heat pain of mice 3h after administration, and have certain analgesic activity.

Drawings

FIG. 1 Effect of T1910 extract on the rate of increase of mechanical pain threshold in the sole of the foot in mice (Mean + -SD, n-12)

FIG. 2 Effect of T1910 extract on the increase in incubation time of light and heat pain in mice after 1h of administration^*P<0.05, comparison with blank group (Mean ± SD, n ═ 12)

FIG. 3 Effect of T1910 extract on the increase in incubation time of light and heat pain in mice after 3h administration^*P<0.05, comparison with blank group (Mean ± SD, n ═ 12)

Detailed Description

The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.

The raw material medicines (or medicinal materials) adopted by the invention can be purchased from common pharmaceutical shops or Chinese medicinal material selling companies, and the specification of the raw material medicines (or medicinal materials) accords with the national pharmaceutical standard or the relevant regulations of Chinese pharmacopoeia and the like. The adopted medicinal materials are Chinese medicinal decoction pieces unless otherwise specified, and the Chinese medicinal decoction pieces can also be obtained and processed.

The curative effect standard of the invention refers to the relevant curative effect standard in the 'Chinese medicine disease judgment curative effect standard'. And (3) healing: clinical symptoms all disappeared and the laboratory check was normal. Improvement: clinical symptoms were reduced, laboratory examinations were improved or normal. And (4) invalidation: the clinical symptoms are not obviously improved or aggravated.

The preparation process of the invention uses the technical requirements of the research on the preparation process of new traditional Chinese medicines in principle, uses the new technology of modern preparations to extract the main effective components of the medicines for use, and adds some pharmaceutically acceptable auxiliary materials or carriers.

Example 1 preparation of herbal extracts

The T1910 medicinal material is unique in the basic source plants and is distributed in southern Shaanxi, southern Gansu and northern Sichuan. Growing in mountain sparse forest with elevation of 1100-3700 m. The root-like stem and root are used as the medicine by Han nationality, Uygur nationality and other nationalities.

Preparation of T1910 extract

The experimental process comprises the following steps: crushing the T1910 medicinal material, adding 8 times of solvent into the medicinal material powder, heating in a water bath until the medicinal material powder is boiled, carrying out heat preservation and reflux extraction for 1 hour, and filtering while the medicinal material powder is hot; adding 6 times of solvent into the filter residue, repeating the reflux extraction operation, reflux extracting for 1 hour, filtering while hot, combining the filtrates, concentrating and volatilizing the solvent to obtain crude extract. The information is as follows:

TABLE 1 different ethanol extract information table

Numbering	Extraction solvent	Weight of T1910 medicinal material	Extract weight g	Yield of
					Frantion 1	Water (W)	80g	24	30％
Frantion 2	30% ethanol	125g	35	28％
					Frantion 3	50% ethanol	200g	50	25％
Frantion 4	80% ethanol	250g	55	22％

Distribution of T191080% ethanol extract ethyl acetate-n-butanol-water system

The experimental process comprises the following steps: crushing the T1910 medicinal material, adding 80% ethanol in an amount which is 8 times that of the medicinal material powder, heating in a water bath until the medicinal material powder is boiled, carrying out heat preservation and reflux extraction for 1 hour, and filtering while the medicinal material powder is hot; adding 80% ethanol 6 times the amount of the filter residue, repeating the reflux extraction operation, performing reflux extraction for 1 hour, and filtering while the solution is hot; mixing filtrates, concentrating, and removing solvent to obtain crude extract. Adding 550ml purified water, extracting with ethyl acetate twice (250 ml/time), mixing ethyl acetate phases, concentrating and volatilizing solvent to obtain ethyl acetate part; extracting the water phase with water saturated n-butanol twice (250 ml each time), mixing n-butanol phases, concentrating, and volatilizing solvent to obtain n-butanol fraction; the remaining aqueous phase was concentrated. The information is as follows:

TABLE 280% ethanol extract solvent distribution information table

Example 2T 1910 extract analgesic Activity screening

The current animal pain model building method is to cause pain reaction by specific stimulation to experimental animals, such as light, heat, mechanical stimulation and the like, and to evaluate the effect of the drug on the pain reaction of the animals. Wherein, the mechanical pain sensitivity method and the light-heat pain sensitivity method are two commonly adopted experimental methods. On the basis of preliminary investigation on the mechanical pain plantar stimulation effect, tail tip thermal pain sensitivity in a tail flick illumination experiment and the drug effect time, in the experiment, a acupuncture plantar tester is adopted to measure the avoidance force of a mouse, a tail flick pain tester is used to measure the avoidance latency, the analgesic effect of the T1910 extract on the mouse is evaluated by two methods of mechanical pain stimulation and thermal stimulation, and analgesic active parts are screened.

Method and device

1. Animal grouping and administration

240 SPF grade ICR mice, male, with a body weight of 23-26 g. After 3d of adaptive feeding, the animals were randomly divided into a blank control group, a Fr1 (Low/Medium/high) group, a Fr2 (Low/Medium/high) group, a Fr3 (Low/Medium/high) group, a Fr4 (Low/Medium/high) group, a Fr5 (Low/Medium/high) group, and a Fr6 (Low/Medium/high) group, and 12 animals were administered in the doses and groups as detailed in Table 3. All the medicines are dissolved by 0.05 percent of CMC-Na solution in an ultrasonic mode, after the pelma pain threshold value and the tail flick illumination latency period baseline are measured respectively for each mouse, the mice are continuously administrated with the stomach irrigation for 4 days, and the blank group and the model group are administrated with 0.05 percent of CMC-Na solution with the same volume. Measuring the avoidance force of Kunming mice by using an electronic Von Frey pain measuring instrument after administration for 3h on the third day, measuring the avoidance latency by using a rat tail illumination pain measuring instrument after administration for 1h and 3h on the fourth day, and evaluating the analgesic effect of the extract on the mice by using two methods of mechanical pain stimulation and illumination radiation heat stimulation.

TABLE 3T 1910 series of different extract groupings and dosing regimens

2. Plantar mechanical pain threshold detection

2.1 Baseline survey

According to the administration time sequence, the mouse is sequentially placed in the special organic glass with the latticed bottom, after the application is carried out for 30min, the left foot (central position) of the mouse is detected by using an electronic Von Frey pain tester, the foot lifting of the mouse is taken as a pain response, and the pain threshold value is recorded. All mice were tested for 1 cycle, 5 cycles. The 3 measurements with the smaller deviation from the median were selected, averaged and the percentage increase in plantar mechanical pain threshold was calculated.

2.2 plantar mechanical pain threshold detection after drug administration

After each group of drug treatment, the plantar mechanical pain threshold of each group of mice is detected 3 hours after the drug administration on the third day. All mice were tested for 1 cycle, 3 times, averaged and the percentage increase in plantar mechanical pain threshold calculated. After the detection is finished, the mice should be returned to the cage immediately to ensure the supply of food and water for the mice.

3. Illumination and thermalgia latency detection

3.1 Baseline survey

The mice were fixed in an organic glass sleeve and the test was started after the mice were quiet. Marking the 3cm position of the mouse tail, aligning the position with a thermal radiation light source, starting the instrument, and recording the incubation period of the light thermal pain when the tail flick of the mouse is a pain reaction. All mice were tested for 1 cycle, 3 cycles, averaged and the percentage increase in light-induced thermal pain latency calculated.

3.2 detection of incubation time to light Heat pain after drug administration

After each group was given drug treatment, the incubation period of light-induced thermal pain was examined in each group of mice 1h and 3h after the fourth day of drug administration. All mice were tested for 1 cycle, 3 cycles, averaged and the percentage increase in light-induced thermal pain latency calculated. After the detection is finished, the mice should be returned to the cage immediately to ensure the supply of food and water for the mice.

4. Data analysis

Statistical analysis is carried out on experimental data by adopting GraphPad and SPSS23.0 software, and the result is expressed by Mean value plus or minus standard deviation (Mean plus or minus SD); one-way ANOVA (One way ANOVA) was used, and the difference was statistically significant with P < 0.05.

Second, results and analysis

1. Effect of T1910 extract on mechanical pain threshold in the soles of mice

The rate of increase of mechanical pain threshold of sole of each group of mice after 3h of administration on the third day is shown in fig. 1, compared with the blank group, the mechanical pain threshold of sole of each group of mice has a certain trend of increasing but has no significant difference after administration of different doses of T1910 extract (Fr1-Fr 6). Among the mice in the Fr2 medium/high dose, Fr3 high dose and Fr6 high dose groups, the increase rate of the mechanical pain threshold value of the soles is obvious, but the individual difference is large, and the statistical significance is not achieved.

2. Effect of T1910 extract on incubation period of mouse light-induced thermal pain

The increase rate of the incubation period of phototoxic lighting after 1h and 3h administration on day four in each group of mice is shown in fig. 2 and fig. 3. As can be seen from the figure, the administration time 1h was constant for each administration group. Wherein, the increase rate of the phototoxic incubation period of mice in Fr1 high dose, Fr3 medium/high dose, Fr4 low/medium/high dose and Fr6 high dose groups is obviously increased.

TABLE 4 mechanical pain threshold and increase rate of tail flick light incubation (Mean + -SD, n is 12) for each group of mice sole

Note:^**P<0.01，^*P<0.05, comparison with blank group

After administration for 3h, the tail-flick phototaxis pain incubation period of Fr1 high-dose and Fr3 medium/high-dose drug group mice is obviously increased compared with that of blank group mice (P is less than 0.05); the tail flick phototoxic incubation period of mice in Fr4 medium/high dose and Fr6 high dose groups also has a certain rising trend, but the individual difference is large, so the statistical significance is avoided.

The results show that the Fr1 high dose and the Fr3 medium/high dose have certain analgesic activity when being administrated for 3 hours, can obviously improve the tail flick phototoxic pain incubation period of mice, and does not influence the independent activity of the mice. In addition, after the Fr6 high-dose group mice are administrated, the incubation period of light-induced thermal pain and the mechanical pain threshold increasing rate both have obvious rising trends after 3 hours, but no obvious difference exists, and the obvious rising trends are probably related to the lower yield and the lower administration dose in the tested medicament.

Third, conclusion

The high dose Fr1 and the medium/high dose Fr3 can obviously improve the incubation period of light and heat pain of mice 3h after administration, and have certain analgesic activity.

Example 3

In order to further understand the efficacy of extracting chopsticks, experiments were designed respectively:

experiment one: the experimental process comprises the following steps: crushing the T1910 medicinal material, adding 6 times of solvent into the medicinal material powder, heating in a water bath until the medicinal material powder is boiled, carrying out heat preservation and reflux extraction for 1 hour, and filtering while the medicinal material powder is hot; adding 4 times of solvent into the filter residue, repeating the reflux extraction operation, reflux extracting for 1 hour, filtering while hot, combining the filtrates, concentrating and volatilizing the solvent to obtain crude extract.

Experiment two: the experimental process comprises the following steps: crushing the T1910 medicinal material, adding 6 times of solvent into the medicinal material powder, heating in a water bath until the medicinal material powder is boiled, carrying out heat preservation and reflux extraction for 1 hour, and filtering while the medicinal material powder is hot; adding 4 times of solvent into the filter residue, repeating the reflux extraction operation, reflux extracting for 1 hour, filtering while hot, combining the filtrates, concentrating and volatilizing the solvent to obtain crude extract.

Experiment three: the experimental process comprises the following steps: crushing the T1910 medicinal material, adding 10 times of solvent into the medicinal material powder, heating in a water bath until the medicinal material powder is boiled, carrying out heat preservation and reflux extraction for 1 hour, and filtering while the medicinal material powder is hot; adding 8 times of solvent into the filter residue, repeating the reflux extraction operation, reflux extracting for 1 hour, filtering while hot, combining the filtrates, concentrating and volatilizing the solvent to obtain crude extract.

Experiment four: the experimental process comprises the following steps: crushing the T1910 medicinal material, adding 10 times of solvent into the medicinal material powder, heating in a water bath until the medicinal material powder is boiled, carrying out heat preservation and reflux extraction for 1 hour, and filtering while the medicinal material powder is hot; adding 8 times of solvent into the filter residue, repeating the reflux extraction operation, reflux extracting for 1 hour, filtering while hot, combining the filtrates, concentrating and volatilizing the solvent to obtain crude extract.

The extracts obtained by the preparation are respectively used for investigating the analgesic effect on mice, and the results show that the extracts have similar effect.

The above examples are intended to illustrate the disclosed embodiments of the invention and are not to be construed as limiting the invention. In addition, various modifications of the methods and compositions set forth herein, as well as variations of the methods and compositions of the present invention, will be apparent to those skilled in the art without departing from the scope and spirit of the invention. While the invention has been specifically described in connection with various specific preferred embodiments thereof, it should be understood that the invention should not be unduly limited to such specific embodiments. Indeed, various modifications of the above-described embodiments which are obvious to those skilled in the art to which the invention pertains are intended to be covered by the scope of the present invention.

Claims

1. Use of radix Chimonanthi Praecocis extract in preparing analgesic medicine is provided.

2. The use as claimed in claim 1, wherein the extract of Chimonanthus praecox is obtained by extracting Chimonanthus praecox with an extraction solvent.

3. The use as claimed in claim 1, wherein the extract of Chimonanthus praecox is prepared by adding 6-10 times of solvent into the raw materials, heating, refluxing, filtering, adding 4-8 times of solvent into the residue, refluxing, filtering, mixing filtrates, and concentrating.

4. An analgesic drug is characterized in that the effective component of the drug is an extract of Chimonanthus praecox.

5. The medicament of claim 4, wherein: the extract of Chimonanthus praecox is prepared by extracting Chimonanthus praecox with an extraction solvent.

6. The medicament of claim 5, wherein: the preparation method of the radix Chimonanthi Praecocis extract comprises adding 6-10 times of solvent into the raw materials, heating, refluxing, filtering, adding 4-8 times of solvent into the filter residue, refluxing, extracting, filtering, mixing the filtrates, and concentrating.

7. The medicament of claim 6, wherein: the preparation method comprises adding 8 times of solvent into the medicinal materials, heating to boil, refluxing for 1h, filtering while hot, adding 6 times of solvent into the filter residue, refluxing for 1h, filtering, mixing filtrates, and concentrating into extract.

8. The medicament of claim 4, wherein: the concentration of the Chimonanthus praecox extract in the medicine is 20-80 mug/ml.

9. The medicament of claim 5, wherein: the solvent is water or ethanol with the volume fraction of 30-50%.

10. An analgesic drug combination is characterized by comprising a therapeutically effective amount of an extract of Chimonanthus praecox and at least one other analgesic drug.