CN114306333A

CN114306333A - Application of Niaoling in preparing medicine for treating arthritis

Info

Publication number: CN114306333A
Application number: CN202111653056.3A
Authority: CN
Inventors: 朱丽君; 刘中秋; 李小翠; 倪家栋; 许仪红; 傅愉; 区晓君
Original assignee: Guangzhou University of Traditional Chinese Medicine
Current assignee: Guangzhou University of Traditional Chinese Medicine
Priority date: 2021-12-30
Filing date: 2021-12-30
Publication date: 2022-04-12

Abstract

The invention discloses application of Niaoling in preparing a medicament for treating arthritis, and can also be applied in preparing medicaments for treating arthritis and having anti-inflammatory and analgesic activity effects, and rat experiments prove that the Niaoling has obvious treatment effect on rheumatoid arthritis, can obviously reduce the concentration of MMP3 in serum, has obvious treatment effect superior to that of indometacin, and has no side effect; the experiment proves that the traditional Chinese medicine composition has the characteristics of obvious anti-inflammatory and analgesic activities, high permeability, good absorption, high oral bioavailability and the like, has good pharmacy, good medicinal prospect and can enrich the treatment means of the rheumatoid arthritis.

Description

Application of Niaoling in preparing medicine for treating arthritis

Technical Field

The invention relates to the field of medical biotechnology, in particular to application of a traditional Chinese medicine active ingredient, namely Niaoling which can be used for treating rheumatoid arthritis.

Background

Rheumatoid Arthritis (RA) is a chronic, inflammatory, systemic autoimmune disease. Clinically, it is characterized by symmetrical swelling of the joints, morning stiffness, local joint pain, dysfunction, and associated mild or moderate fever. RA has high disability rate, and the erosion of multi-joint cartilage and bone can occur when the treatment is not timely carried out, thereby seriously affecting the life quality of patients. The etiology and pathogenesis of RA are not completely clear, and the current treatment of RA mainly aims at controlling symptoms and delaying disease conditions. The western medicine treatment of RA mainly takes non-steroidal anti-inflammatory drugs, glucocorticoids, immunosuppressants and the like as main materials, so that adverse reactions cannot be ignored while the disease condition is delayed. In recent years, scholars at home and abroad make extensive and intensive research on the aspect of treating RA by using plant medicines, and find that a plurality of natural medicinal components such as erycibe glycoside, total glucosides of paeony, poison vine and the like have better RA treatment effect. Therefore, the safe and efficient RA treatment medicine has wide prospect from the botanical medicine.

Niolanlin (Neoline, NE) is C-19 type aconite alkaloid extracted and separated from traditional Chinese medicine monkshood, the current research on NE at home and abroad mainly focuses on extraction and separation and structure identification, the research on the pharmacological activity and application of NE is very little, the known research shows that NE has the effects of strengthening heart, dilating blood vessels, resisting tumors and the like in vitro, and the NE has the application in resisting arrhythmia, heart failure, pain and the like, but the research report of the NE for treating the rheumatoid arthritis is not available at present.

Disclosure of Invention

Therefore, based on the background, the invention provides the application of Niaoling (CAS number: 466-26-2) in preparing the medicine for treating arthritis, and a new means can be provided for treating rheumatoid arthritis through the research on the new application of Niaoling.

The technical scheme of the invention is as follows:

one of the invention points of the invention is that: use of NE in the manufacture of a medicament for the treatment of arthritis; the structural formula of the NE is as follows:

further, the arthritis is rheumatoid arthritis.

Furthermore, the dosage form of the medicine is a conventional dosage form for clinically treating arthritis.

Further, the dosage form of the medicine can be oral preparation, injection preparation or external absorption preparation.

The invention also provides a medicament for resisting rheumatoid arthritis, which comprises effective dose of NE and pharmaceutically acceptable auxiliary agents.

The anti-rheumatoid arthritis medicament is an oral preparation or an injection preparation or an external preparation, wherein the oral preparation type comprises but is not limited to tablets, capsules, granules and the like.

The invention also provides the application of NE in the preparation of medicaments with anti-inflammatory, analgesic and antipyretic activities.

By adopting the technical scheme, the method has the following beneficial effects:

the rat experiment proves that NE has obvious curative effect on rheumatoid arthritis, can obviously reduce the concentration of Matrix metalloproteinase-3 (MMP3) which is an arthritis marker in serum, and has obvious better curative effect than indometacin; the experiment proves that the traditional Chinese medicine composition has the characteristics of obvious anti-inflammatory, analgesic and pyrolytic activities, and the like, also has the advantages of high permeability, good absorption, high oral bioavailability and the like, has better pharmacy property, good medicinal prospect and can enrich the treatment means of the rheumatoid arthritis.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.

FIG. 1 is a photograph of the swelling and lesion status of the right hind paw of a rat in the experiment of NE in example 1 of the present invention for treating RA;

wherein the Normal group (Normal) (1A), the Model group (Model) (1B), the Positive drug group (Positive) (1C), the high dose NE group (4mg/kg NE) (1D) and the low dose NE group (2mg/kg NE) (1E).

FIG. 2 is a graph of the joint and foot swelling rate scores at

days

9, 12, 15, 18 and 21 for rats in the normal group, model group, positive drug group, high dose NE group and low dose NE group in the experiment of treatment of RA with NE of example 1 of the present invention;

FIG. 3 is a graph of the joint swelling score on

days

9, 12, 15, 18 and 21 in rats of the normal group, model group, positive drug group, high dose NE group and low dose NE group in the experiment of NE in example 1 of the present invention for treating RA;

FIG. 4 is the arthritic index scores at

days

9, 12, 15, 18 and 21 for rats in the normal group, model group, positive group, high dose NE group and low dose NE group in the experiment of NE in example 1 of the present invention for the treatment of RA;

FIG. 5 is a graph of the systemic scores of rats in the normal group, model group, positive drug group, high dose NE group and low dose NE group on

days

9, 12, 15, 18 and 21 in the experiment of treatment of RA with NE of example 1 of the present invention;

FIG. 6 is a graph showing the body weight changes of rats in the normal group, model group, positive drug group, high-dose NE group and low-dose NE group on

days

9, 12, 15, 18 and 21 in the experiment of treating RA with NE of example 1 of the present invention;

FIG. 7 is a graph showing the concentrations of matrix metalloproteinase 3(MMP3) in the sera of rats of the normal group, model group, positive drug group, high-dose NE group and low-dose NE group in the experiment of treating RA with NE of example 1 of the present invention;

FIG. 8 is a graph showing the swelling rate of the hind paw of a mouse injected with lambda carrageenan for 4 hours after the mouse was perfused with Saline (50 mg/kg), diclofenac sodium (Positive), NE (15 mg/kg) and NE (7.5 mg/kg) for 30min, respectively, in example 2 of the present invention;

FIG. 9 shows the number of pain and sprain times after intraperitoneal injection of 1% acetic acid after respectively gavage with Saline (Saline), diclofenac sodium (50 mg/kg), NE (15 mg/kg) and NE (7.5 mg/kg) for 30min in mice in example 2 of the present invention;

FIG. 10 shows the recorded heat-sensitive threshold values of the mice in example 2 of the present invention displayed on the liquid crystal panel when the mice were subjected to IR55 infrared radiation with intensity in the right hind paw after being infused with Saline (Saline), diclofenac sodium (50 mg/kg), NE (15 mg/kg) and NE (7.5 mg/kg) for 30min, and when the mice had leg-lifting, beating and foot-licking actions;

FIG. 11 is a blood concentration-time curve of mice of example 3 of the present invention after oral administration of 15mg/kg NE (FIG. 11A) and intravenous injection of 1.5mg/kg NE (FIG. 11B), respectively;

FIG. 12 is the pharmacokinetic parameters of oral administration of 15mg/kg NE and intravenous injection of 1.5mg/kg NE in mice of example 3 of the present invention:

wherein FIG. 12A is the area under the plasma concentration-time curve (AUC)_0-t)；

FIG. 12B is the Mean Residence Time (MRT)_0-t)；

FIG. 12C is half-life (T)_1/2)；

FIG. 12D is the apparent distribution volume (V)_d)；

Fig. 12E is Clearance (CL);

FIG. 13 is a graph showing the uptake and transport characteristics of NE of example 3 of the present invention in a Caco-2 monolayer cell model:

wherein FIG. 13A is the effect of efflux transporter P-gp inhibitor (Verapamul), BCRP inhibitor (KO143) and MRP2 inhibitor (MK571) on the amount of transport of NE from the AP-BL side;

FIG. 13B is a graph of the effect of 3 efflux transporter inhibitors on AP-side transport of NE from BL-AP side;

FIG. 13C is the apparent permeability coefficients (P) for NE for 3 efflux transporter inhibitors_app) The influence of (a);

figure 13D is the effect of 3 efflux transporter inhibitors on the efflux rate of NE.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1: experiment on treatment of RA with NE

This example is a therapeutic experiment of main Niaoling (NE) on rat adjuvant arthritis to investigate the therapeutic effect of NE on rheumatoid arthritis.

30 male SD rats (120. + -.20 g) were used, and 6 rats were randomly divided into 5 groups, i.e., a normal group, a model group, an indomethacin group (positive drug), a 4mg/kg NE group (high dose NE group), and a 2mg/kg NE group (low dose NE group). After the rats were fixed, the hair around the root of the tail was shaved off, wiped with 75% alcohol for sterilization, and 0.1mL of physiological saline was injected subcutaneously at about 1cm from the root of the tail of the normal group of rats, while the other groups of rats were injected with the same volume of Freund's complete adjuvant. The onset started on day 9 after injection of Freund's complete adjuvant in rats. Rats in the normal group, model group, indomethacin group, 4mg/kg NE group and 2mg/kg NE group were gavaged with 20% beta-cyclodextrin, 2mg/kg indomethacin, 4mg/kg NE and 2mg/kg NE, respectively, starting on day 9, and were gavaged continuously for 14 days. In the experiment, 2mg/kg of indomethacin, 4mg/kg of NE and 2mg/kg of NE are prepared by using 20% beta-cyclodextrin. After the onset of disease from the 9 th day rat, the paw volume and body weight of the rat were measured and recorded every 3 days, and the number of joint swelling, arthritis score and whole body score of the rat were calculated.

Calculating the foot swelling rate of the rat:

joint swelling counts 1 wrist ankle (or ankle) and 5 finger (toe) joints per paw, with a maximum of 24 joints swelling per rat.

The arthritis index scoring standard is as follows:

recording 0 point when the standard is normal;

erythema and slight swelling of the ankle joint, scoring 1;

erythema and slight swelling appeared from ankle joint to toe joint or metacarpal joint, and 2 points were recorded;

erythema and moderate swelling appeared from ankle to toe or metacarpal joints, and 3 points are recorded;

erythema and severe swelling occurred in the ankle to toe joints, and was scored 4.

The whole body scoring standard is as follows:

the ear had no tubercle and redness, and scored 0;

one ear with a tubercle and redness scored 1;

nodules in both ears and redness, score 2;

the nose has no connective tissue swelling, and the score is 0;

obvious connective tissue swelling is marked as 1 point;

the tail is without nodules and is marked with 0 point;

1 point is marked if there is a nodule;

the paw is not swollen, and 0 point is recorded;

swelling of one forefoot, score 1;

swelling of the two forefoot paws, 2 points are recorded;

the hindpaw has no swelling and is scored as 0;

swelling of one hind paw, score 1;

swelling of the two hind paws, 2 points are recorded; each rat was rated a maximum of 8 points.

After the experiment is finished, fixing the rats, and shooting the right hind paw pictures of the rats in each group; the rats were then anesthetized and blood was taken through the abdominal aorta, and after the blood sample was allowed to stand at room temperature for 2 hours, serum was separated by centrifugation at 3000rpm for 10min, and the concentration of Matrix metalloproteinase-3 (Matrix metalloproteinase-3, MMP3) as an arthritis marker in the serum of each rat was measured according to the instructions of the Elisa kit. The results obtained in this experiment are shown in FIGS. 1 to 7. Wherein the results of figures 2-7 are expressed as means ± standard deviation, the data are analyzed using the Shapiro-Wilk test for normal distribution, for two sets of mean-compared data, the data are analyzed using the independent sample t-test for normal distribution and homogeneity of variances, and the data are analyzed using the satterhwaite corrected t' test for irregular variances. In addition, data were analyzed using nonparametric Wilcoxon rank sum test when they did not fit into a normal distribution. P <0.05, indicates a statistical difference. In the figure, "#" indicates that there is a statistical difference between the model group and the normal group, and "#" indicates that there is a statistical difference between the drug-treated group and the model group (n ═ 6).

According to the analysis of the results in fig. 1 to 7, the onset of disease in rats started on the 9 th day of molding. The swelling, redness and deformation of the joints were evident in the model group rats (fig. 1B) compared to the normal group rats (fig. 1A); from day 9 onwards, the foot swelling rate score (fig. 2), joint swelling number score (fig. 3), arthritis index score (fig. 4) and whole body score (fig. 5) were all increased in the model group rats compared with the normal group; and from day 12 onwards, the foot swelling rate score (fig. 2), joint swelling number score (fig. 3), arthritis index score (fig. 4) and whole body score (fig. 5) of the model group rats were significantly increased compared to the normal group (P < 0.05). Meanwhile, the body weight of the model group rats began to decrease from day 9 (fig. 6), and the body weight of the model group rats was significantly lower than that of the normal group rats from day 12 (P < 0.05). In addition, the serum concentration of MMP3 (arthritis marker) in rats in the model group was also significantly increased compared to the normal group (fig. 7, P < 0.05). The results show that the rat adjuvant arthritis model is successfully established.

As is apparent from fig. 1 to 7, NE has a significant effect in treating adjuvant arthritis in rats. Compared with the model group, after the rats of the high-dose NE group and the low-dose NE group are treated by NE, the joint swelling, redness and deformation of the rats are obviously improved, and the improvement degree is equivalent to that of the indometacin treatment group (figure 1); both NE and indomethacin treatment reduced the paw swelling rate score of adjuvant arthritic rats compared to the model group (figure 2), and from day 12, high dose (4mg/kg) NE treatment significantly reduced the paw swelling rate of adjuvant arthritic rats (P <0.05), and low dose (2mg/kg) NE treatment also significantly reduced the paw swelling rate of adjuvant arthritic rats (P <0.05) on days 12 and 15; both NE and indomethacin treatment decreased the joint swelling score of adjuvant arthritic rats compared to the model group (figure 3), and both NE and indomethacin treatment at high/low dose from day 12 significantly decreased the joint swelling score of adjuvant arthritic rats (P < 0.05); both NE and indomethacin treatment reduced the arthritis index score of adjuvant arthritis rats compared to the model group (figure 4), and high/low dose NE treatment significantly reduced the arthritis index score of adjuvant arthritis rats on days 12, 15 and 18 (P < 0.05); both high/low dose NE and indomethacin treatment significantly reduced systemic scores in adjuvant arthritis rats compared to the model group (figure 5, P < 0.05); both high/low dose NE and indomethacin treatment significantly restored body weight in adjuvant arthritis rats compared to the model group (figure 6, P < 0.05); both high/low dose NE and indomethacin treatment significantly reduced the concentration of MMP3 in the serum of adjuvant arthritis rats compared to the model group (fig. 7, P < 0.05). In conclusion, the NE has a remarkable effect of treating RA, not only can remarkably improve the joint swelling, redness and deformation conditions of an adjuvant arthritis rat, but also can remarkably reduce the foot swelling rate score, the joint swelling number score, the arthritis index score and the systemic score of the adjuvant arthritis rat, and can remarkably recover the weight of the adjuvant arthritis rat, and simultaneously remarkably reduce the concentration of MMP3 in the serum of the adjuvant arthritis rat.

Example 2: pharmacological and pharmacodynamic related experiment of NE for treating RA

The main clinical features of RA are arthritic swelling, localized joint pain and associated mild or moderate fever. Therefore, anti-inflammatory, analgesic and antipyretic effects are key to the relief of RA symptoms, and are effective in eliminating joint swelling, alleviating pain and restoring body temperature. The experiment proves the pharmacological activity of NE in resisting inflammation, easing pain and relieving heat.

(1) Anti-inflammatory activity of NE

This example examines the anti-inflammatory activity of NE using a classical carrageenan-induced mouse foot swelling inflammation model. Male 24C 57 mice at 8 weeks of age were selected, randomized into 4 groups of 6, and the thickness of the right hind paw of each mouse was measured with a vernier caliper. After the 4 groups of mice were each perfused with 10mL/kg of physiological saline, 50mg/kg of diclofenac sodium (positive drug), 15mg/kg of NE and 7.5mg/kg of NE for 30min, 20. mu.L of lambda carrageenan was subcutaneously injected to the right plantar aspect of each group of mice to cause inflammation. After 4h of inflammation, the thickness of the right foot of each mouse was measured with a vernier caliper. The foot swelling rate of each mouse was calculated using the following formula:

the anti-inflammatory assay results for NE are shown in figure 8, with the results expressed as mean ± standard deviation, the data analyzed using independent sample t-test, and the symbol "×" indicates P <0.05, i.e. significant differences between the drug-treated group and the saline group (n ═ 6). As shown in fig. 8, 50mg/kg diclofenac sodium and 15 and 7.5mg/kg NE significantly reduced the foot swelling rate of the mice 58.95%, 56.02% and 70.20%, respectively (P <0.05), compared to the saline group. The lambda carrageenan-induced inflammation model is a classic anti-inflammatory drug screening model, and experimental results show that NE has obvious anti-inflammatory activity, and the anti-inflammatory activity of the NE is equivalent to that of diclofenac sodium which is a non-steroidal anti-inflammatory drug. The significant anti-inflammatory activity of NE is probably one of the drug effect bases of NE in treating RA, and can significantly inhibit the joint inflammation of RA patients.

(2) Analgesic action of NE

The invention adopts a classical acetic acid induced mouse pain writhing model to investigate the analgesic activity of NE. Male 8-week-old C57 mice, 24, were randomly divided into 4 groups of 6 mice each. After the 4 groups of mice are respectively perfused with the normal saline, 50mg/kg of diclofenac sodium (positive drug), 15mg/kg of NE and 7.5mg/kg of NE for 30min according to the volume of 10mL/kg, 1% acetic acid solution is intraperitoneally injected according to the volume of 10mL/kg, the number of times of pain and wriggle of the mice is recorded after 5min, and the total 15min is observed. The pain inhibition rate of each group of medicines is calculated through the times of writhing, and the calculation formula is as follows:

the results of the NE analgesia assay are shown in fig. 9, with the results expressed as mean ± standard deviation, the data analyzed by t-test on independent samples, and the symbol ".", indicates P <0.05, i.e. there was a significant difference between the drug-treated group and the saline group (n ═ 6). Compared with the normal saline group, 50mg/kg of diclofenac sodium, 15mg/kg of NE and 7.5mg/kg of NE can obviously reduce the number of painful writhing of mice (P is less than 0.05), and the pain inhibition rates are 45.44%, 68.18% and 41.82% respectively. The acetic acid induced mouse pain writhing model is a classic analgesic screening model, and experimental results show that NE has a remarkable analgesic effect, and the analgesic activity of the NE is equivalent to that of diclofenac sodium, a non-steroidal anti-inflammatory drug. Therefore, analgesic activity may be one of the drug bases of NE in the treatment of RA, and can significantly relieve the symptoms of arthralgia in patients with RA.

(3) Antipyretic action of NE

The invention adopts a classical infrared thermosensitive experimental model to investigate the antipyretic activity of NE. Male 8-week-old C57 mice, 24, were randomly divided into 4 groups of 6 mice each. After the 4 groups of mice are respectively perfused with the normal saline, 50mg/kg diclofenac sodium (positive drug), 15mg/kg NE and 7.5mg/kg NE for 30min according to the volume of 10mL/kg, infrared radiation is given to the right hind paw of the mice, and the infrared light intensity is IR 55. And recording the mouse heat-sensitive threshold displayed by the liquid crystal display screen after the mouse acts like leg lifting, flapping, foot adding and the like. In order to avoid scalding the mice, the detection of the mice without leg raising reaction is stopped after 25s, and the threshold value is calculated according to 25 s.

The results are shown in fig. 10, the results are expressed as mean ± standard deviation, the data are analyzed by t-test on independent samples, and the symbol "×" indicates P <0.05, i.e. there is a significant difference between the drug-treated group and the saline group (n ═ 6). Compared with the saline group, 50mg/kg diclofenac sodium, 15mg/kg NE and 7.5mg/kg NE can respectively increase the thermal threshold value of the mice by 188.86%, 95.65% and 118.75% (P < 0.05). The infrared thermosensitive experiment is a common antipyretic screening model, and the experimental result shows that NE has obvious antipyretic effect. Therefore, the antipyretic activity of NE is probably one of the drug effect bases of NE in treating RA, and can remarkably recover the body temperature of RA patients.

Example 3: oral bioavailability and absorption transport characteristics of NE

The present implementation initially evaluates the potency of NE by examining its oral bioavailability and absorption transport characteristics.

(1) Oral bioavailability of NE

Male 8-week-old C57 mice, 10, were randomized into 2 groups of 5 mice each. 2 groups of mice were perfused with gastric lavage 15mg/kg NE and intravenously injected with 1.5mg/kg NE, and blood was collected from the tail vein at

time points

0, 3, 7, 10, 15, 20, 30, 50, 80, 120, 240, 360 and 480 min. After blood samples are centrifugally separated to obtain plasma, protein is precipitated by an organic solvent, 2uL of the plasma is taken to enter a UHPLC-MS/MS system for quantitatively detecting the concentration of NE in the plasma, a blood concentration-time curve is drawn, pharmacokinetic parameters are calculated, and the oral bioavailability is calculated according to the following formula:

oral bioavailability (100%) (AUC)_po×Dose_iv)/(AUC_iv×Dose_po)×100％

(2) Uptake transport characteristics of NE

Caco-2 cells in logarithmic growth phase were diluted with DMEM complete medium containing 10% fetal bovine serum, 1% diabody and 1% nonessential amino acids, seeded in 6-well Transwell plates to a cell density of 50 ten thousand/well, and cultured in a 5% CO2 incubator at 37 ℃ for 21 days. Changing the solution once a day during the cell culture period, after the cells grow into a stable and compact monolayer cell tissue for 21 days, washing the cell tissue with HBSS buffer solution at 37 ℃ for 3 times, measuring the trans-epithelial cell resistance, and when the resistance value is more than 420 omega/cm 2, indicating that a compact and complete cell monolayer is formed and can be used for a transmembrane transport experiment. After the resistance measurement is finished, 2mL of HBSS buffer solution is added to two sides of each hole, and the mixture is placed on a shaking table at 37 ℃ for incubation for 30min, so that the cells release nutrients absorbed in the culture process. Drug transport process from AP side to BL side: 2mL of HBSS solution containing 2. mu.M NE was added from the AP side, and an equal volume of blank HBSS solution was added to the BL side. 0.5mL of each sample was sampled on both sides at 30, 60, 90 and 120min after administration, while 0.5mL of HBSS solution containing 2. mu.M NE was supplemented on the AP side and an equal volume of blank HBSS solution was supplemented on the BL side, and each set of samples was divided into 3 portions. The transport of drug from BL side to AP side was the same as the transport experiment from AP side to BL side, i.e. drug was added to BL side and blank HBSS solution was added to AP side.

Inhibitor experiments: when the drug is transported from AP-BL, adding a mixed HBSS solution containing 2 μ M NE and an inhibitor from the AP side, and simultaneously adding a blank HBSS solution from the B side; when the drug is transported from BL-AP, the drug is added to the BL side, the HBSS solution containing the inhibitor is added to the A side, and the rest steps are the same as above. And adding 100 mu L of acetonitrile solution containing an internal standard into the sample, mixing uniformly, centrifuging, and taking 2 mu L of supernatant to perform UHPLC-MS/MS analysis to detect the concentration of NE.

The results are shown in fig. 11 to 13, where the results are expressed as mean ± standard deviation, the data are analyzed by t-test on independent samples, and the symbol ".", indicates P <0.05, i.e. there is a significant difference (n ═ 6) between the different inhibitor groups and the control group. The results of in vivo pharmacokinetic experiments show that NE is rapidly absorbed and has high absorption degree in mice, the blood concentration reaches the peak (figure 11A) within 12min, and the oral bioavailability is 63.82%. Apparent permeability coefficient (P) of NE in vitro cell transport experiments_app) Mainly at 2.1 x 10^-5cm/s to 4.59 x 10^-5In the cm/s range (FIG. 13C), it was shown that the NE is well absorbed on the cells, probably entering and exiting the cells mainly by passive diffusion; in addition, the Efflux rates of NE on Caco-2 cells were all significantly reduced when inhibitors of Efflux transporters P-gp, BCRP and MRP2 were added (FIG. 13D, P <0.05), indicating that Efflux transporters P-gp, BCRP and MRP2 are all involved in the Efflux of NE, but the Efflux rate of NE is very low (Efflux ratio < 2.14). In conclusion, NE has good oral absorption, high bioavailability and good drug action, and can be applied to the preparation of drugs for treating RA.

Through the experiments of examples 1 to 3, it can be obviously seen that NE can remarkably treat and improve the symptoms of adjuvant arthritis of rats, and the treatment effect of NE is superior to that of indomethacin which is a non-steroidal anti-inflammatory drug; through a pharmacodynamic basic experiment, NE has excellent anti-inflammatory activity, analgesic activity and antipyretic activity, and the excellent anti-inflammatory activity, analgesic activity and antipyretic activity can be obviously improved and relieved symptoms of arthritis; the experiments of oral bioavailability and in vitro absorption and transportation prove that the NE has good oral absorption, low discharge rate, high bioavailability and better druggability.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

Claims

1. The application of the Niaoling in preparing the medicine for treating the arthritis is characterized in that the Niaoling has the following structural formula:

2. the use according to claim 1, wherein the arthritis is rheumatoid arthritis.

3. The use of claim 1, wherein the medicament is in a dosage form that is conventional for the clinical treatment of arthritis.

4. The use according to claim 3, wherein the medicament is in the form of an oral preparation, an injectable preparation or an external preparation.

5. The medicine for treating rheumatoid arthritis is characterized by comprising effective amount of Niaoling and pharmaceutically acceptable auxiliary agents.

6. The drug according to claim 5, wherein the drug is in a dosage form of oral preparation, injection preparation or external preparation.

7. Application of Niaoling in preparing medicine with antiinflammatory, analgesic and antipyretic effects is provided.