Nipailol granules as well as preparation method and application thereof

Abstract

The invention provides a preparation method of nipallol granules, which comprises the following steps: soaking nipallol for 0.5-2.5 hours by using 10-14 times by weight of water, decocting for 0.5-1.5 hours for 1-5 times, collecting filtrate, and concentrating to obtain extract; uniformly mixing the nipallol extract with auxiliary materials to prepare a soft material, granulating and drying to obtain nipallol granules; the weight ratio of the Nipailuo extract to the auxiliary materials is (1: 1)-(1: 2); the auxiliary material consists of glucose and soluble starch; the weight ratio of the glucose to the soluble starch is (1: 1)-(3: 7). The nipallol granules provided by the invention can remarkably inhibit the mouse auricle swelling rate (Plt; 0.05), and has a significant anti-inflammatory effect; the average painful body twisting frequency (Plt; 0.05) and the inhibition rate of pain and body twisting, and the inhibition rate is as high as 49.11%; the traditional Chinese medicine composition has a good antidiarrheal effect on mouse diarrhea caused by folium sennae; the traditional Chinese medicine composition has a good curative effect on lamb diarrhea, and the total effective rate is 96.67%.

Description

Technical Field

[0001] This invention belongs to the field of veterinary drug technology, specifically relating to a Nialo granule, its preparation method, and its application. Background Technology

[0002] Lamb diarrhea is a common disease in livestock farming. Due to the incomplete development of the intestinal mucosa in newborn lambs, the incidence is high in lambs during the lambing and lactation periods. The main clinical characteristic is persistent diarrhea. The primary causes of lamb diarrhea are pathogenic microbial infection and digestive system dysfunction, posing a significant threat to sheep farming. Numerous studies have shown that the main pathogens causing lamb diarrhea are Gram-negative bacteria such as Escherichia coli, Salmonella, Clostridium perfringens, Clostridium perfringens, and Shigella.

[0003] Currently, the main treatments for pathogenic microbial infections are antibiotics or chemical drugs used alone or in combination. However, with the increasing drug resistance of various pathogens, such as Escherichia coli, the main pathogen, the development of new treatments is urgent. Among these, developing new veterinary drugs from natural plants such as Tibetan medicine has become one of the best options.

[0004] Nialuo is the whole herb of two Polygonaceae plants, Polygonum difformis and Polygonum aviculare (forked Polygonum), which grow on mountain slopes, river terraces, grassy slopes at the edges of shrublands, and sand dunes at altitudes of 3000–4900 m. It has a slightly bitter, sour, and astringent taste, and is cool in nature. This invention systematically studies the pharmacological effects of Nialuo granules and aims to develop a new Tibetan veterinary drug preparation for treating lamb diarrhea, which has excellent prospects for veterinary clinical application. Summary of the Invention

[0005] To address the problems existing in the prior art, this invention provides Nialo granules, their preparation method, and applications. Through anti-inflammatory, analgesic, antidiarrheal, and preclinical pharmacodynamic studies in target animals, the pharmacological functions of Nialo granules were systematically studied. This invention holds promise for developing a new Tibetan veterinary drug formulation for treating lamb diarrhea, demonstrating excellent prospects for veterinary clinical application.

[0006] The first objective of this invention is to provide a method for preparing Nialo granules, comprising soaking Nialo in 10-14 times its weight of water for 0.5-2.5 hours, then decocting for 0.5-1.5 hours, repeating the decoction 1-5 times, collecting the filtrate and concentrating it into an extract; mixing the Nialo extract with excipients to form a soft mass, granulating and drying it to obtain Nialo granules; wherein the weight ratio of the Nialo extract to the excipients is 1:1-1:2; wherein the excipients are composed of glucose and soluble starch; wherein the weight ratio of glucose to soluble starch is 1:1-3:7.

[0007] As a preferred method, Nialo is soaked in 10 times its weight of water for 1 hour, then decocted for 1.5 hours, decocted 3 times, and the filtrate is collected and concentrated into an extract.

[0008] As a preferred method, Nialo is soaked in 14 times its weight of water for 1 hour, then decocted for 1.5 hours, decocted 4 times, and the filtrate is collected and concentrated into an extract.

[0009] Preferably, the weight ratio of the nialo extract to the excipients is 1:1.

[0010] Preferably, the weight ratio of glucose to soluble starch is 3:7.

[0011] The second objective of this invention is to provide a Nialo particle prepared using the method described above.

[0012] A third objective of this invention is to provide the use of Nialo granules in the preparation of anti-inflammatory, analgesic, or antidiarrheal medications.

[0013] Preferably, the Niallo granules are used in the preparation of a medicine for treating lamb diarrhea.

[0014] Preferably, the dosage of the drug is 0.3797 g / kg to 1.5188 g / kg; more preferably, the dosage of the drug is 0.7594 g / kg to 1.5188 g / kg; and most preferably, the dosage of the drug is 1.5188 g / kg.

[0015] A fourth objective of this invention is to provide a medicine whose active ingredient includes the aforementioned Niallo granules.

[0016] This invention optimizes and screens multiple factors in the extraction method of Niallo, and obtains a method with the highest extraction and preparation efficiency. The optimal ratio of extract to excipients for granule formulation screening is 1:1, and the excipients are glucose and soluble starch with an addition ratio of 3:7. The resulting granules have the best forming rate and solubility, and have the advantages of high yield, low cost, safety and environmental protection, and simple operation, making them suitable for large-scale production.

[0017] The Nialo granules of this invention can significantly inhibit the ear swelling rate induced by xylene in mice (P<0.05), exhibiting significant anti-inflammatory effects; they can significantly reduce the average number of painful writhing movements in mice (P<0.05) and the inhibition rate of painful writhing movements, with an inhibition rate as high as 49.11%; they have a good antidiarrheal effect on senna-induced diarrhea in mice; preclinical pharmacodynamic tests in target animals show that Nialo granules have a good curative effect on lamb diarrhea, with a total effective rate of 96.67%, and are expected to be further developed into a new Tibetan medicine veterinary preparation for treating lamb diarrhea. Attached Figure Description

[0018] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used in conjunction with embodiments of the invention to explain the invention and do not constitute a limitation thereof. In the drawings:

[0019] Figure 1 The results are obtained from the HPLC analysis of the Nialo water extract sample.

[0020] Figure 2 It is a granular formulation with three excipients in a specific ratio. Detailed Implementation

[0021] The following examples are provided to better understand the present invention, but do not limit the invention. Unless otherwise specified, the experimental methods in the following examples are conventional methods. Unless otherwise specified, the experimental materials used in the following examples were purchased from conventional biochemical reagent companies. The quantitative experiments in the following examples were all performed in triplicate, and the results were averaged. Example 1: Optimization Experiment of Nialo Extraction Method

[0022] 1. Single-factor optimization experiment

[0023] The effective components of nialool were extracted using water extraction. The number of extractions (1, 2, 3, 4, 5), extraction time (0.5h, 1h, 1.5h, 2h, 2.5h), material-to-liquid ratio (1:6, 1:8, 1:10, 1:12, 1:14), and soaking time (0.5h, 1h, 1.5h, 2h, 2.5h) were selected as the main factors for single-factor optimization experiments.

[0024] According to HPLC analysis, the aqueous extract of Nialo was found to be rich in gallic acid. Figure 1 The study proposes to use gallic acid content as an evaluation index to determine the content of nialo extract samples by single-factor analysis.

[0025] Figure 1 The results are obtained from the HPLC analysis of the Nialo water extract sample.

[0026] HPLC determination method: An Agilent SB-C18 column was used. The mobile phase was methanol-0.1% (weight / volume percentage, i.e., 0.1% means that 0.1g of phosphoric acid (solute) is contained in 100mL of solution) phosphoric acid aqueous solution (volume ratio 10:90). The flow rate was 0.8mL / min, the detection wavelength was 273nm, the column temperature was 30℃, and the injection volume was 10μL.

[0027] Sample preparation: Take 100 μL of nialool single-factor water extract with a concentration of 1 g / mL, and dilute to a final volume of 5 mL with 50% methanol. Sonicate for 40 min. Pass the treated sample solution through a 0.22 μm syringe filter to obtain the sample solution for instrumentation.

[0028] (1) Single-factor analysis of extraction frequency

[0029] The number of decoctions (1, 2, 3, 4, and 5) were selected for investigation, with a material-to-liquid ratio of 1:10, an extraction time of 1.5 h, and a soaking time of 1 h for sample preparation. The gallic acid content in the samples was detected by HPLC, and the results are shown in Table 1. With the increase in the number of extractions, the gallic acid content in the decoction also increased, with significant increases observed at 2 and 4 extractions. This indicates that too few decoctions will result in incomplete extraction of the active ingredients from the medicinal material, while too many extractions will increase economic costs. Therefore, 2, 3, and 4 extractions were ultimately selected as the three levels for investigation in the orthogonal experiment.

[0030] Table 1. HPLC results of single-factor evaluation of extraction number.

[0031] Sample Name Peak area Gallic acid content (mg / g) Extract once 635795 0.642 Extract 2 times 1088433 1.099 Extract 3 times 1222695 1.235 Extract 4 times 1600899 1.616 Extract 5 times 1736408 1.753

[0032] (2) Extraction time single factor examination

[0033] Extraction times of 0.5 h, 1 h, 1.5 h, 2 h, and 2.5 h were selected for investigation. The material-to-liquid ratio was 1:10, extraction was performed three times, and samples were prepared by soaking for 1 h. The gallic acid content in the samples was detected by HPLC, and the results are shown in Table 2. With increasing extraction time, the gallic acid content in the extract also increased. The increase in gallic acid content was more significant when the extraction time increased from 0.5 h to 1 h, and then from 1 h to 1.5 h. Considering that excessively long extraction times would lead to energy waste, extraction times of 0.5 h, 1 h, and 1.5 h were ultimately selected as the three levels for investigation in the orthogonal experiment.

[0034] Table 2. HPLC results of single-factor evaluation of extraction time.

[0035] Sample Name Peak area Gallic acid content (mg / g) Extraction for 0.5 hours 1325331 1.338 Extraction 1h 1512623 1.527 Extraction time: 1.5 hours 1763352 1.780 Extraction 2h 1728582 1.745 Extraction time 2.5 hours 1816230 1.834

[0036] (3) Single-factor analysis of the extraction liquid-to-material ratio

[0037] The extraction solid-liquid ratios (by weight) of 1:6, 1:8, 1:10, 1:12, and 1:14 were selected for investigation. The extraction time was 1.5 h, with three extractions and a 1 h soaking time for sample preparation. Gallic acid content in the samples was determined by HPLC, and the results are shown in Table 3. As the extraction solid-liquid ratio increased, the gallic acid content in the decoction also increased. Based on literature review and practical experience, the commonly used extraction solid-liquid ratio is generally between 1:10 and 1:20. However, excessively high solid-liquid ratios waste energy; therefore, solid-liquid ratios of 1:10, 1:12, and 1:14 were selected as three levels for investigation in the orthogonal experiment.

[0038] Table 3. HPLC results of single-factor investigation of extract-liquid ratio.

[0039] Sample Name Peak area Gallic acid content (mg / g) The ratio of material to liquid is 1:6 684303 0.691 The ratio of material to liquid is 1:8 1079204 1.090 Material-to-liquid ratio 1:10 1223373 1.235 The ratio of material to liquid is 1:12 1294174 1.307 The ratio of material to liquid is 1:14 1508781 1.523

[0040] (4) Single-factor analysis of soaking time

[0041] Soaking times of 0.5h, 1h, 1.5h, 2h, and 2.5h were selected for investigation. Extraction time of 1.5h, three extractions, and a material-to-liquid ratio of 1:10 were used for sample preparation. Gallic acid content in the samples was detected by HPLC, and the results are shown in Table 4. The gallic acid content did not change significantly with the change in soaking time, indicating that the soaking time has little effect on the extraction process. Following the commonly used soaking time for medicinal materials, a soaking time of 1h was selected for subsequent experiments.

[0042] Table 4. HPLC results of single-factor evaluation of soaking time

[0043]

[0044]

[0045] 2. Orthogonal optimization experiment

[0046] Based on the single-factor experiments, three factors were ultimately selected for investigation: extraction times, extraction time, and extraction-to-liquid ratio. For each factor, three levels were selected for L9(3) experiments. 4 Orthogonal design was used to screen for the optimal extraction process. The factors and levels of the orthogonal experiment are shown in Table 5.

[0047] Table 5. Factor Level Table for Orthogonal Experiment

[0048]

[0049] Gallic acid content and dry extract yield were used as comprehensive evaluation indicators. A multi-indicator total evaluation normalization method was employed for statistical analysis, and the calculation formula is as follows:

[0050] Total score normalized value (OD) = (d1 + d2 + d3 + ... + dk) / k (where k is the number of indicators)

[0051] di = (Yi - Ymin) / (Ymax - Ymin) (Ymin is the minimum value of the indicator, and Ymax is the maximum value of the indicator)

[0052] The method for determining the yield of the extract is to take 20 mL of orthogonal extraction sample into an evaporating dish, place it in an oven at 105℃ and dry it to constant weight, and then calculate the yield.

[0053] The results of the orthogonal experiment are shown in Table 6. The order of importance of the three factors is: number of extractions > extraction time > extraction material-liquid ratio. Therefore, the optimal extraction scheme is A3B3C3, which means adding 14 times the amount of water and decocting 4 times for 1.5 hours each time.

[0054] Table 6. Results of the intuitive analysis of the orthogonal experiment.

[0055]

[0056]

[0057] The results of the analysis of variance are shown in Table 7. The results show that the number of extractions, extraction time, and extraction-to-liquid ratio have no significant effect. Considering the experimental results and economic costs, the extraction process parameters of Nialo are appropriately adjusted, and the extraction process is proposed to be optimized to A1B3C2, that is: add 10 times the amount of water, decoct 3 times, 1.5 hours each time.

[0058] Table 7. Results of Variance Analysis of Orthogonal Experiments

[0059] factor Sum of squared deviations Degrees of freedom F ratio F critical value Significance Extraction liquid ratio 0.096 2 3.000 19.000 Number of extractions 0.488 2 15.250 19.000 Extraction time 0.257 2 8.031 19.000 error 0.03 2

[0060] 3. Process Validation and Optimization

[0061] Three batches of medicinal materials were weighed in parallel. Based on the optimal and proposed optimized extraction schemes determined by orthogonal experiments, extraction process verification experiments were conducted, and the results are shown in Table 8. The results show that when the extraction material-to-liquid ratio and the number of extractions were reduced, the differences in gallic acid content and dry extract yield were relatively small. Considering the need to improve extraction efficiency, reduce production costs, and enhance economic benefits, the final extraction process was determined to be A1B3C2, namely: adding 10 times the amount of water, decocting three times, each time for 1.5 hours.

[0062] Table 8 Results of the Extraction Process Validation Test

[0063]

[0064] 4. Summary of Extraction Process

[0065] Based on single-factor investigation, orthogonal optimization experiment and optimal process verification study, the extraction process of Nialool was optimized by comprehensively considering extraction efficiency and economic cost. The process is as follows: weigh Nialool according to the prescription, add 10 times the amount of water, soak for 1 hour, decoct for 1.5 hours, decoct 3 times, and combine the decoctions to obtain Nialool extract.

[0066] II. Nialo Granule Preparation Process Experiment

[0067] Preparation of Niallo Extract: According to the determined optimal extraction process, accurately weigh appropriate amounts of Niallo, pulverize them, soak for 1 hour, add 10 times the weight of the medicinal material in water, and extract for 1.5 hours each time, for a total of 3 extractions. Combine the extracts and concentrate to a Niallo extract concentration of 1 g / mL.

[0068] Excipient selection: Four excipients—glucose, sucrose, soluble starch, and dextrin—were selected for preliminary experiments, as the type of excipient is a crucial factor affecting granulation formation. Based on the preliminary experimental results, glucose and soluble starch were ultimately chosen as the granulation excipients. The 2020 Chinese Veterinary Pharmacopoeia, Part II (Appendix 8), requires that the amount of excipients generally not exceed twice the amount of drug. Therefore, drug-to-excipient weight ratios of 1:1, 1:1.5, and 1:2 (with a fixed glucose:soluble starch weight ratio of 1:1) were proposed for excipient formulation. The granulation quality was evaluated by comprehensively considering the granulation rate and solubility of the three formulations; the results are shown in Table 9.

[0069] Molding rate determination: Weigh the prepared granules, pass them through a No. 1 sieve, then through a No. 5 sieve, collect the granules that can pass through the No. 1 sieve but not the No. 5 sieve, and weigh them. Molding rate = (mass of granules after sieving / mass of granules before sieving) × 100%.

[0070] Solubility test: Accurately weigh 5g of the prepared granules, add 100mL of boiling water, stir and shake for 5min, and observe whether it can be completely dissolved.

[0071] Table 9. Evaluation of Particle Formation Rate and Solubility for Three Different Proportions

[0072] sample Molding rate Solubility 1:1 granules 89.72％ Completely dissolved 1:1.5 particles 89.67％ Completely dissolved 1:2 granules 88.17％ Completely dissolved

[0073] The results showed that the granules prepared by the three ratios of 1:1, 1:1.5, and 1:2 all had good solubility and met the relevant requirements for granules in Part II (Appendix 8) of the 2020 edition of the Chinese Veterinary Pharmacopoeia, with little difference in the formation rate. Considering the ease of the granulation process, a drug-to-excipient ratio of 1:1 was ultimately selected.

[0074] The ratio of glucose and soluble starch as excipients was further determined. The drug-to-excipient ratio was fixed at 1:1, and granulation was planned with glucose to soluble starch weight ratios of 5:5, 4:6, and 3:7. Figure 2 The resulting particles were also examined for their formation rate and solubility, and the results are shown in Table 10.

[0075] Figure 2 It is a granular formulation with three excipients in a specific ratio.

[0076] Table 10: Evaluation of Particle Formation Rate and Solubility of Auxiliary Material Proportions

[0077] sample Molding rate Solubility Glucose: Soluble starch 5:5 89.36％ Completely dissolved Glucose: Soluble starch 4:6 88.61％ Completely dissolved Glucose: Soluble starch 3:7 90.93％ Completely dissolved

[0078] The results showed that the granules prepared with the three excipient ratios all had good solubility and met the relevant requirements for granules in the 2020 edition of the Chinese Veterinary Pharmacopoeia, Part II (Appendix 8). The granules with a 3:7 ratio of two excipients had a higher formation rate than the other two ratios. Therefore, the final production process for the granules was determined as follows: the drug to excipient ratio was 1:1, the ratio of glucose to soluble starch was 3:7, the nialool extract was mixed with the excipients to form a soft mass, granulated through a No. 1 sieve, dried at 60℃, and then granulated through a No. 1 and No. 5 sieve to obtain nialool granules.

[0079] III. Pharmacodynamic trials of Nialo Granules for anti-inflammatory, analgesic, and antidiarrheal effects

[0080] 1. Anti-inflammatory test

[0081] Fifty Kunming mice were randomly divided into five groups: a low-dose Nialo granule group, a medium-dose Nialo granule group, a high-dose Nialo granule group, a positive control group, and a model control group, with ten mice in each group. Half were male and half were female. The volume of drug administered by gavage to each group was calculated as 0.2 mL / 20 g·BW. The Nialo granule dosages were 0.3797 g / kg, 0.7594 g / kg, and 1.5188 g / kg, respectively. The positive control group (aspirin) dosage was 0.2600 g / kg. The model control group was administered an equal volume of deionized water by gavage. Administration was repeated once daily for 3 days. The granules were prepared to the prescribed concentration and administered at the same volume based on body weight. Thirty minutes after administration on day 3, 20 μL of 100% xylene solution was precisely pipetted and quickly applied to both the front and back of the right ear of each mouse to induce inflammation. The left ear was left untreated. Mice were sacrificed 30 minutes after inflammation induction. Ear pieces were punched at the same location on the left and right ears of mice using a puncher. The pieces were then weighed on a precision electronic balance, and their wet weight was recorded. The degree of ear swelling, swelling rate, and the drug's inhibition rate of ear swelling were calculated. The statistical results are shown in Table 11.

[0082] Mouse ear swelling degree = right ear wet mass - left ear wet mass.

[0083] Mouse ear swelling rate = (wet mass of right ear - wet mass of left ear) / wet mass of left ear × 100%.

[0084] The drug's inhibition rate of mouse ear swelling = (mean ear swelling in the model control group - mean ear swelling in the drug group) / mean ear swelling in the model control group × 100%.

[0085] Table 11 Results of the effect of Nialo granules on xylene-induced ear swelling in mice

[0086] Group Auricular swelling rate % Inhibition rate % Nialo granules low-dose group 71.32±6.71 14.34 Nialo granules medium dose group 57.44±16.01** 31.00 Nialo granule high-dose group 47.47±10.85** 42.98 Positive control group 42.37±11.38** 49.10 Model control group 83.26±5.56 -

[0087] The results showed that high-dose and medium-dose Nialo granules inhibited mouse ear swelling by 42.98% and 31.00%, respectively, and significantly inhibited xylene-induced ear swelling in mice, demonstrating significant anti-inflammatory effects.

[0088] 2. Analgesia test

[0089] Fifty Kunming mice were randomly divided into five groups: a low-dose Nialo granule group, a medium-dose Nialo granule group, a high-dose Nialo granule group, a positive control group, and a model control group, with ten mice in each group. Half were male and half were female. The volume of drug administered by gavage to each group was calculated as 0.2 mL / 20 g·BW. The Nialo granule dosages were 0.3797 g / kg, 0.7594 g / kg, and 1.5188 g / kg, respectively. The positive control group (Changyan Ning) dosage was 0.90 g / kg. The model control group was administered an equal volume of deionized water by gavage. The mice were administered the drugs once daily for three consecutive days. Sixty minutes after administration on day 3, each group of mice was intraperitoneally injected with 1% glacial acetic acid at 10 mL / kg BW. Immediately after injection, the mice were placed in a special observation box, and the latency period of pain writhing and the number of times the mice writhing in pain within 20 minutes were recorded in real time. The pain writhing inhibition rate was calculated, and the results are shown in Table 12.

[0090] Mouse pain writhing inhibition rate = (average number of pain writhing in the model group - average number of pain writhing in the drug treatment group) / average number of pain writhing in the model group × 100%.

[0091] Table 12 Results of the effects of Nialo granules on the latency and number of writhing movements induced by glacial acetic acid in mice (n=10)

[0092] Group Painful writhing latency / s Average number of painful twisting movements Painful writhing inhibition rate % Nialo granules low-dose group 104.5±41.14 31.5±6.02* 20.25 Nialo granules medium dose group 134.1±45.47 28.3±3.74** 28.35 Nialo granule high-dose group 201.85±32.03 20.1±7.45** 49.11 Positive control group 217.9±20.67 21.9±4.81** 44.56 Model control group 120.56±32.27 39.5±4.36 -

[0093] The results showed that, compared with the model group, the high-dose Nialo granules group could prolong the latency of pain writhing in mice, and Nialo granules in each dose group could significantly reduce the average number of pain writhing in mice (P<0.01 or P<0.05). Moreover, as the dose of Nialo granules increased, the inhibition rate of pain writhing in mice also increased significantly, reaching a maximum of 49.11%.

[0094] 3. Antidiarrheal test

[0095] Fifty Kunming mice were randomly divided into six groups: a low-dose Nialo granule group, a medium-dose Nialo granule group, a high-dose Nialo granule group, a positive control group, a model control group, and a blank control group, with 10 mice in each group (half male and half female). An acute diarrhea model was established in mice by gavage for 5 consecutive days, with senna leaf at a concentration of 1 g / mL (calculated as 0.2 mL / 20 g BW). After successful model establishment, the gavage volume of the drug in each group was calculated as 0.2 mL / 20 g BW, and the Nialo granule dosages were 0.3797 g / kg, 0.7594 g / kg, and 1.5188 g / kg, respectively. The positive control group was gavage with an equal volume of Pulsatilla chinensis oral liquid, while the model control group and the blank control group were gavage with an equal volume of deionized water. The administration was repeated for 3 consecutive days, once daily. Feces from each group were collected and weighed daily from 9:00 AM to 2:00 PM. After drying in an oven, the feces were weighed again and the water content of each experimental group was calculated. The results are shown in Table 13.

[0096] Table 13 Results of the effect of Nialo granules on fecal water content in mice (n=10)

[0097] Group Modeling 5D fecal moisture content / g Stool water content after 3 days of administration / g low-dose group 2.18±0.85** 1.02±0.27 medium dose group 2.31±0.47** 0.99±0.89 High-dose group 2.10±0.63** 0.57±0.51 Positive control group 2.27±0.71** 0.48±0.29 Model control group 2.03±0.50** 2.52±0.72** Blank control group 0.34±0.05 0.37±0.04

[0098] The results showed that 5 days after establishing the acute diarrhea model, the fecal water content of mice in other groups was significantly different from that in the blank control group (P<0.01), indicating successful modeling. 3 days after administration, there was no significant difference in fecal water content between the low-dose, medium-dose, and high-dose groups of Nialool and the blank control group (P>0.05), indicating that Nialool granules have a good antidiarrheal effect.

[0099] IV. Preclinical Efficacy Studies in Target Animals

[0100] 1. Materials

[0101] 1.1 Experimental Animals

[0102] To further clarify the therapeutic effect of Nialuo granules, preclinical efficacy trials of Nialuo granules were conducted in six townships in Ngari Prefecture of Tibet and Zhangye Prefecture of Gansu Province, collecting a total of 210 cases of lambs with obvious diarrhea symptoms.

[0103] 1.2 Drugs

[0104] Nialo particles were prepared according to the final method determined in step two.

[0105] 2 methods

[0106] 2.1 Inclusion criteria for experimental animals

[0107] Diagnostic criteria: Characterized by severe diarrhea, with stools that are porridge-like or watery, yellowish-white, yellowish-green, or grayish-white in color, or with a foul odor; a significant increase in the frequency of bowel movements; accompanied by abdominal distension, abdominal pain, etc.

[0108] Selection criteria: Lambs aged 1 to 30 days, without obvious signs of dehydration, and sick lambs can suckle on their own.

[0109] Exclusion criteria: Lambs with other complications, such as pneumonia, omphalitis, and arthritis, or those that have already received other treatments, were not included in this study.

[0110] Exclusion criteria: Lambs that received anti-inflammatory or antibacterial treatments unrelated to this study or failed to complete the prescribed dosing regimen during the trial should be excluded from the final results analysis.

[0111] 2.2 Administration Method

[0112] The granules are dissolved in water at a concentration of 2 g / kg and administered orally for 3 consecutive days.

[0113] 2.3 Criteria for Evaluating Therapeutic Effect

[0114] Cure: The sick lambs regained their normal spirit and appetite, their feces were granular, and there was no loose feces on their tails, buttocks and hind legs. The lambs did not show diarrhea symptoms again within 48 hours after the diarrhea stopped.

[0115] Effective: The sick lambs showed some improvement in spirit and appetite, and their feces became slightly thicker, although there was still loose feces on their tails, rumps and hind legs.

[0116] Ineffective: The symptoms have not improved or have worsened.

[0117] Death: The condition worsens abnormally, leading to death.

[0118] Table 14. Statistical analysis of preclinical efficacy results of Nialo granules in target animals.

[0119]

[0120] As shown in Table 14, the Nialo granules of the present invention can effectively treat lamb diarrhea, with a cure rate of 90.95% and a total effective rate of 96.67%.

[0121] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing Nialo particles, characterized in that: Nialo is soaked in 10-14 times its weight of water for 0.5-2.5 hours, then decocted for 0.5-1.5 hours, 1-5 times. The filtrate is collected and concentrated into an extract. The nialo extract is mixed with excipients to form a soft mass, which is then granulated and dried to obtain nialo granules. The weight ratio of the nialo extract to the excipients is 1:1-1:

2. The excipients consist of glucose and soluble starch, and the weight ratio of glucose to soluble starch is 1:1-3:

7.

2. The method for preparing Nialo particles according to claim 1, characterized in that: Soak Nialo in 10 times its weight of water for 1 hour, then decoct for 1.5 hours. Repeat the decoction three times, collect the filtrate, and concentrate it into an extract.

3. The method for preparing Nialo particles according to claim 1, characterized in that: After soaking Nialo in 14 times its weight of water for 1 hour, it was decocted for 1.5 hours. The decoction was repeated 4 times, and the filtrate was collected and concentrated into an extract.

4. The method for preparing Nialo particles according to claim 1, characterized in that: The weight ratio of the Nialo extract to the excipients is 1:

1.

5. The method for preparing Nialo particles according to claim 1, characterized in that: The weight ratio of glucose to soluble starch is 3:

7.

6. A type of Nialo particle, characterized in that: It is prepared by applying the method described in any one of claims 1-5.

7. The use of the Nialo granules according to claim 6 in the preparation of anti-inflammatory, analgesic or antidiarrheal drugs.

8. The application according to claim 7, characterized in that: The application of the Nialo granules in the preparation of a drug for treating lamb diarrhea.

9. The application according to claim 8, characterized in that: The dosage of the drug is 0.3797 g / kg to 1.5188 g / kg; preferably, the dosage of the drug is 0.7594 g / kg to 1.5188 g / kg; most preferably, the dosage of the drug is 1.5188 g / kg.

10. A drug, characterized in that: Its active ingredient includes the Niallo particles as described in claim 6.

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