Medical Use of Anemoside B4 in the Treatment of Oral Ulcers
20230102973 · 2023-03-30
Inventors
Cpc classification
A61K31/704
HUMAN NECESSITIES
International classification
Abstract
Anemoside B4 is used in the preparation of a medicament for the treatment of oral ulcers. For example, anemoside B4 is used in the preparation of a medicament for the treatment of recurrent oral ulcers or oral ulcers caused by chemotherapy.
Claims
1. A method for treating oral ulcer, comprising administering a medicament comprising anemoside B4 to a subject in need thereof.
2. The method of claim 1, wherein the oral ulcer is a recurrent oral ulcer.
3. of the method of claim 1, wherein the oral ulcer is an oral ulcer caused by chemotherapy.
4. The method according to claim 1, wherein anemoside B4 is a sole active ingredient in the medicament.
5. The method according to claim 1, wherein the medicament comprises anemoside B4 as a first active ingredient and one or more active ingredients selected from the group consisting of rifampicin, dexamethasone, metronidazole, zinc sulfate, montmorillonite, chlorhexidine, chitosan, sucralfate, polyinosinic-polycytidylic acid (poly I:C), levamisole, vitamin C, B vitamins, granulocyte-macrophage colony stimulating factor (GM-CSF), recombinant human epidermal growth factor (RH EGF), and recombinant human keratinocyte growth factor (rh-KGF).
6. The method according to claim 1, wherein the medicament is an oral preparation or a non-oral preparation.
7. The method according to claim 6, wherein the non-oral preparation is selected from the group consisting of injections, rectal preparations, oral mucosa patches, and oral films.
8. The method according to claim 7, wherein the injection is selected from the group consisting of subcutaneous injections, intramuscular injections, and intravenous injections.
9. The method according to claim 7, wherein the rectal preparation is selected from the group consisting of rectal suppositories, rectal gels, and rectal perfusion agents.
10. The method according to claim 1, wherein the subject in need thereof is a mammal or a human.
Description
EXAMPLES
[0021] The present invention will be elucidated below with reference to specific examples. Those skilled in the art will appreciate that these examples are intended to illustrate the present invention only and not to limit the scope of the invention in any way.
[0022] The experimental methods in the following examples, unless otherwise specified, are conventional methods. The starting materials, reagents and the like used in the following examples are commercially available, unless otherwise specified.
Experimental Example 1
Effect of Anemoside B4 on Oral Ulcer Induced by Acetic Acid in Rats
1. The Objective of the Experiment
[0023] The therapeutic effect of anemoside B4 on oral ulcer was evaluated by establishing an acetic acid-induced oral ulcer model in rats and observing the change in food intake and ulcer diameter as well as healing time after the model was established.
2. Experimental Materials
2.1 Experimental Animals
[0024] SFP SD rats, male, 200-220 g, purchased from Hunan SJA Laboratory Animal Co., Ltd., Certificate No. 43004700060832, License No. SOCK (Xiang) 2016-0002.
2.2 Test Drugs
[0025] Anemoside B4 suppository: self-made, 0.14 g/suppository; based on the content of anemoside B4, there are three specifications: 14 mg/suppository, 7 mg/suppository and 3.5 mg/suppository, batch number: 20190523;
[0026] Blank suppository: self-made, prepared in parallel with anemoside B4 suppository, batch number: 20190523-k;
[0027] Xilei Powder: Jiangsu 707 Natural Pharmaceutical Co., Ltd., 190102;
[0028] Dexamethasone: Henan Runhong Pharmaceutical Co., Ltd., Drug Approval Number H41020330.
[0029] The preparation method of anemotide B4 suppository was as follows:
[0030] Anemoside B4 is passed through a 100 mesh sieve for later use; and then the matrix (mixed fatty acid glycerides 36# and 38#) is weighed, melted in a water bath at 60° C., and vigorously stirred, to which was slowly added anemoside B4 under stirring, until anemoside B4 is evenly dispersed in the matrix. The mixture was poured into a plug mold, cooled, and then taken out, to obtain the suppository.
2.3 Experimental Reagents
[0031] Glacial acetic acid (Xilong Science Co., Ltd., batch number 180510), chloral hydrate (Tianjin Damao Chemical Reagent Factory, batch number 20180518), isoflurane (Shenzhen RWD Life Science Co., Ltd., 217180501).
2.4 Experimental Instruments
[0032] Anesthesia machine (RWD, R580), vernier caliper (Mitutoyo, CD-6″CSX).
3. Experimental Method
3.1 Establishment of Rat Model of Oral Ulcer Induced by Acetic Acid
[0033] Rats were anesthetized by intraperitoneal injection of 10% chloral hydrate solution. After anesthesia, 3 mm×3 mm filter paper impregnated with 100% glacial acetic acid was clung to the right buccal mucosa of rats for 45 s. After cauterization for 45 s, a round or oval white lesion of the buccal mucosa in this area was observed, indicating successful modeling.
3.2 Experimental Groups
[0034] All animals with successful modeling were randomly divided into six groups according to the ulcer area: (1) model group, 14 rats; (2) Xilei powder (15 mg/kg) group, 12 rats; (3) dexamethasone (0.3 mg/kg) group, 13 rats; (4) anemoside B4 high dose (about 60 mg/kg*3, i.e. 14 mg/suppository, one suppository/time, 3 times/day) group, 14 rats; (5) anemoside B4 medium dose (about 30 mg/kg*3, i.e. 7 mg/suppository, one suppository/time, 3 times/day) group, 14 rats; (6) anemoside B4 low dose (about 15 mg/kg*3, i.e. 3.5 mg/suppository, one suppository/time, 3 times/day) group, 14 rats; and 6 rats were included in the normal group.
3.3 Administration
[0035] 24 h after modeling, drug was administrated for 3 successive days, including: local administration in Xilei powder group, once a day; intraperitoneal injection in dexamethasone group, once a day; rectal administration in anemoside B4 group, 3 times a day, and administration of blank suppository in model group, 3 times a day.
3.4 Outcome Measures
[0036] (1) General observation: daily food intake per cage was measured from the 2nd day after modeling, and animal weight was measured every 2 days.
(2) Measurement of ulcer surface diameter: on the 2nd, 4th and 5th day after modeling, the diameter of ulcer surface was measured with a vernier caliper and recorded.
4. Experimental Results
4.1 Effect of Anemoside B4 Suppository on Body Weight in Rats with Oral Ulcer
[0037] The results are shown in Table 1.
TABLE-US-00001 TABLE 1 Effect of anemoside B4 suppository on body weight of rats with oral ulcer induced by acetic acid B4 (
As shown in Table 1, the body weight of rats in dexamethasone group decreased significantly after administration. Except for dexamethasone group, the body weight of rats in other groups tended to increase after administration compared with the model group. Since the 4th day (3 days after administration), the body weight of rats in anemoside B4 groups had obviously increased compared with dexamethasone group, with significant difference (p<0.05).
4.2 Effect of Anemoside B4 on Food Intake in Rats with Acetic Acid-Induced Oral Ulcer
[0038] The results are shown in Table 2.
TABLE-US-00002 TABLE 2 Effect of anemoside B4 suppository on food intake in rats with oral ulcer induced by acetic acid (
[0039] As shown in Table 2, after the model was established, the food intake of rats in model group was significantly decreased compared with the normal group, and especially on the 3rd day after modeling, the least food intake was observed in model group and each drug group, indicating that the ulcer was the most severe on the 3rd day, and the food intake of each group was reduced due to pain. Subsequently, as the ulcer healed, the pain decreased and the food intake gradually increased in each group. On the 3rd day after modeling, when the ulcer was the most severe, the food intake of each anemoside B4 group was higher than that of the model group, and the difference between the high and medium dose groups was statistically significant (p<0.05), indicating that the ulcer injury of each anemotide B4 group was mild. The food intake of dexamethasone group increased on the 3rd day (one day after administration) compared with the model group, but then decreased obviously compared with the model group, and was always less than that of anemotide B4 groups.
4.3 Effect of Anemoside B4 on the Diameter of Oral Ulcers Induced by Acetic Acid in Rats
[0040] The results are shown in Table 3.
TABLE-US-00003 TABLE 3 Effect of anemoside B4 suppository on the diameter of oral ulcers induced by acetic acid in rats (
[0041] The data in Table 3 showed that anemoside B4 suppository could significantly reduce the diameter of acetic acid-induced oral ulcers in rats from the 4th and 5th day after modeling, having statistical difference (p<0.05) compared with the model group. Its effect of promoting ulcer healing was comparable to that of dexamethasone and better than that of Xilei powder.
5. Conclusion
[0042] Rectal administration of anemoside B4 had a good therapeutic effect on oral ulcer induced by acetic acid in rats, and its effect of promoting ulcer healing is similar to that of dexamethasone, but significantly better than that of Xilei powder. Meanwhile, anemoside B4 is better than dexamethasone in improving the overall conditions of rats with oral ulcer reflected by body weight and food intake.
Experimental Example 2
Therapeutic Effect of Anemoside B4 on Oral Ulcer Induced by Paclitaxel in Rats
1. The Objective of the Experiment
[0043] Oral ulcer is a common adverse reaction in cancer patients after radiotherapy and chemotherapy. In this experiment, a rat oral ulcer model induced by chemotherapeutic drug paclitaxel was used to evaluate the therapeutic effect of anemoside B4 on oral ulcer by observing the food intake and ulcer healing time of rats after modeling.
2. Experimental Materials
2.1 Experimental Animals
[0044] SFP SD rats, male,180-200 g, purchased from Hunan SJA Laboratory Animal Co., Ltd., Certificate No. 43004700060832, License No. SOCK (Xiang) 2016-0002.
2.2 Test Drugs
[0045] Anemoside B4 suppository: self-made, whose preparation method is the same as that of Example 1; each suppository weights 0.15 g and contains 14 mg of anemoside B4, batch number 20190523.
[0046] Blank suppositories were prepared in parallel.
[0047] Dexamethasone: Henan Runhong Pharmaceutical Co., Ltd., Drug Approval Number H41020330.
2.3 Experimental Reagents
[0048] Paclitaxel injection (Beijing Shuanglu Pharmaceutical Co., Ltd., batch number 20190101), chloral hydrate (Tianjin Damao Chemical Reagent Factory, batch number 20180518), isoflurane (Shenzhen RWD Life Science Co., Ltd., 217180501).
2.4 Experimental Instruments
[0049] Anesthesia machine (RWD, R580), vernier caliper (Mitutoyo, CD-6″CSX).
3. Experimental Method
3.1 Establishment of Rat Model of Oral Ulcer Induced by Paclitaxel Injection
[0050] Rats were anesthetized by intraperitoneal injection of 10% chloral hydrate solution. After anesthesia, the oral ulcer model induced by chemotherapeutic drugs was established by injecting 50 μl of 6 mg/ml paclitaxel injection into the right buccal mucosa of rats.
3.2 Experimental Groups
[0051] All animals with successful modeling were randomly divided into model group, (7 rats), anemoside B4 high dose (about 60 mg/kg*2) group (7 rats), and normal group (6 rats).
3.3 Administration
[0052] 24 h after modeling, in anemoside B4 group, rectal administration was performed, 1 suppository/time, twice a day, for 5 successive days; in dexamethasone group, intraperitoneal administration was performed, once a day, for 5 successive days; and in model group, blank suppository was given.
3.4 Outcome Measures
[0053] (1) General observation: On the day of modeling, rats were fasted but free access to water for 12 hours, while the diet was given quantitatively on the second day after modeling, and the food intake of rats in each cage was measured daily from the third day after modeling (the second day after administration).
(2) Measurement of ulcer surface diameter: from the 2nd day after modeling, the diameter of ulcer surface was measured with a vernier caliper and recorded for 7 successive days.
4. Experimental Results
4.1 Effect of Anemoside B4 on Food Intake in Rrats with Paclitaxel-Induced Oral Ulcer
[0054] The results are shown in Table 4.
TABLE-US-00004 TABLE 4 Effect of anemoside B4 on food intake in rats with oral ulcer induced by paclitaxel (
[0055] As shown in Table 4, after the model was established, the food intake of the animals in each model group was significantly decreased compared with that of the normal group, in which the food intake was the least on the 3rd day, then gradually recovered, and basically returned to normal from the 6th day, that meant the food intake was equivalent to that of the normal group. During this period, the food intake of rats in anemoside B4 group was consistently higher than that in the model group, and the difference was statistically significant on days 3 and 4 (p<0.05). However, there was no significant improvement in food intake of dexamethasone group.
4.2 Effect of Anemoside B4 on the Diameter of Oral Ulcers Induced by Paclitaxel in Rats
[0056] The results are shown in Table 5.
TABLE-US-00005 TABLE 5 Effect of anemoside B4 on the diameter of oral ulcers induced by paclitaxel in rats (
[0057] The experimental data in Table 5 indicated that anemoside B4 suppository could significantly reduce the ulcer surface after administration, having a significant difference (P<0.05) compared with the model group from the 3rd day. There was no significant difference in ulcer diameters between dexamethasone group and model group.
5. Conclusion
[0058] The above results showed that anemoside B4 had a good therapeutic effect on paclitaxel-induced oral ulcer in rats, could significantly shorten the healing time of the ulcer, and could improve the overall conditions of rats (food intake increased compared with the model group). The above study suggested that anemoside B4 could be used to treat oral ulcers caused by chemotherapeutic drugs in cancer patients, which could reduce patients' pain, improve their quality of life, and enable chemotherapy to be successfully completed.