CN116983315A - Application of 12-ketolithocholic acid in preparation of medicines for preventing and treating ulcerative colitis - Google Patents
Application of 12-ketolithocholic acid in preparation of medicines for preventing and treating ulcerative colitis Download PDFInfo
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- CN116983315A CN116983315A CN202310736796.6A CN202310736796A CN116983315A CN 116983315 A CN116983315 A CN 116983315A CN 202310736796 A CN202310736796 A CN 202310736796A CN 116983315 A CN116983315 A CN 116983315A
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- ketolithocholic acid
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- CVNYHSDFZXHMMJ-VPUMZWJWSA-N 12-ketolithocholic acid Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)C(=O)C1 CVNYHSDFZXHMMJ-VPUMZWJWSA-N 0.000 title claims abstract description 55
- 206010009900 Colitis ulcerative Diseases 0.000 title claims abstract description 54
- 201000006704 Ulcerative Colitis Diseases 0.000 title claims abstract description 54
- CVNYHSDFZXHMMJ-UHFFFAOYSA-N 12-keto-lithocholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(=O)C2 CVNYHSDFZXHMMJ-UHFFFAOYSA-N 0.000 title claims abstract description 53
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- 229920003045 dextran sodium sulfate Polymers 0.000 description 13
- TVZRAEYQIKYCPH-UHFFFAOYSA-N 3-(trimethylsilyl)propane-1-sulfonic acid Chemical compound C[Si](C)(C)CCCS(O)(=O)=O TVZRAEYQIKYCPH-UHFFFAOYSA-N 0.000 description 12
- 239000007864 aqueous solution Substances 0.000 description 9
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- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical compound C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 description 1
- BHQCQFFYRZLCQQ-UHFFFAOYSA-N (3alpha,5alpha,7alpha,12alpha)-3,7,12-trihydroxy-cholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 BHQCQFFYRZLCQQ-UHFFFAOYSA-N 0.000 description 1
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- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
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- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
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Abstract
The application discloses an application of 12-ketolithocholic acid in preparing a medicament for preventing and treating ulcerative colitis, and belongs to the technical field of biological medicines. The 12-ketolithocholic acid can obviously reduce the weight loss of the ulcerative colitis model mouse, and is beneficial to weight stabilization; can obviously improve diarrhea, loose stool and even fecal occult blood conditions of ulcerative colitis model mice, and reduce disease activity index; can obviously inhibit the shortening of the colon and restore the length of the colon; can obviously relieve colon tissue pathological injury of ulcerative colitis mice; can obviously reduce the ILC3s level of the inherent immune cells of the colon and inhibit the cells from secreting the proinflammatory cytokine IL-17A, thereby playing the role of preventing and treating the exacerbation of ulcerative colitis. The medicine for preventing and treating ulcerative colitis has definite medicinal curative effect, low side effect and wide medical application prospect.
Description
Technical Field
The application belongs to the technical field of biological medicines, and particularly relates to application of 12-ketolithocholic acid in preparation of a medicine for preventing and treating ulcerative colitis.
Background
Ulcerative colitis persists and is even treated for life, which is one of the refractory diseases listed by the world health organization. The number of onset of UC is continuously increased in the last 10 years, and the onset of UC is a bottleneck problem which causes national economic burden. At present, medicines such as 5-aminosalicylic acid, glucocorticoid, immunosuppressant and the like are commonly used as medicines for clinically treating ulcerative colitis.
Clinical application and observation report for many years show that the medicines are easy to relapse after stopping medicines, have adverse reactions of different degrees of hepatorenal toxicity, hormone resistance and the like, have high price, have poor patient compliance and the like, and influence the life quality of patients.
Disclosure of Invention
In order to solve the problems in the prior art, the application aims to provide an application of 12-ketolithocholic acid in preparing a medicament for preventing and treating ulcerative colitis.
The application is realized by the following technical scheme:
application of 12-ketolithocholic acid in preparing medicine for preventing and treating ulcerative colitis is provided.
Preferably, the medicament is for use in increasing body weight, decreasing disease activity index, restoring colon length and decreasing histopathological damage in ulcerative colitis mice.
The above index about recovery of colon length is due to the fact that the colon length of the mice suffering from ulcerative colitis is shortened, and the colon length of the mice gradually tends to be in a normal length state after administration of the drug, and there is a process of recovery of the colon length.
Preferably, the disease activity index comprises the rate of weight loss, diarrhea status and hematochezia status of ulcerative colitis mice.
A medicine for preventing and treating ulcerative colitis takes 12-ketolithocholic acid as an active ingredient.
Preferably, the medicament also contains pharmaceutically acceptable auxiliary materials, and is prepared into a pharmaceutically corresponding dosage form.
Preferably, the HPLC purity of the 12-ketolithocholic acid is not less than 95%.
Preferably, the auxiliary material is a liquid, solid or semisolid auxiliary material.
Preferably, the dosage form includes any one of a liquid dosage form, a solid dosage form, and a semi-solid dosage form;
the liquid dosage forms include solutions and/or injections; the solid dosage forms include powders and/or tablets; the semi-solid dosage forms include suppositories and/or pastes.
Compared with the prior art, the application has at least the following technical effects:
the application provides an application of 12-ketolithocholic acid in preparing a medicament for preventing and treating ulcerative colitis, wherein the 12-ketolithocholic acid can obviously reduce the weight loss of a mouse with an ulcerative colitis model, and is beneficial to weight stabilization; can obviously improve diarrhea, loose stool and even fecal occult blood conditions of ulcerative colitis model mice, and reduce disease activity index; can obviously inhibit the shortening of the colon and restore the length of the colon; can obviously relieve colon tissue pathological injury of ulcerative colitis mice; can significantly reduce the level of the colon innate immune cell-3 type innate lymphoid cell (Group 3innate lymphoid cells,ILC3s) and inhibit the secretion of the proinflammatory cytokine IL-17A by the cell, thereby playing the role of preventing and treating the exacerbation of ulcerative colitis.
The medicine for preventing and treating ulcerative colitis has definite medicinal curative effect, low side effect and wide medical application prospect.
Drawings
FIG. 1 is a graph showing the comparison of the weight change of mice in each group in the effect of the example 12-ketolithocholic acid on the weight of ulcerative colitis mice;
FIG. 2 is a graph showing the comparative changes in disease activity index of various groups of mice in the influence of the example 12-ketolithocholic acid on the disease activity index of ulcerative colitis mice;
FIG. 3 is a graph showing the colon length measurement of each group of mice in the effect of the example 12-ketolithocholic acid on the colon length of ulcerative colitis mice;
FIG. 4 is a graph showing the comparison of colon length of each group of mice in the effect of the example 12-ketolithocholic acid on colon length of ulcerative colitis mice;
FIG. 5 is a microscopic schematic view of the 10X 10 times magnification of the histopathology of each group of mice in the effect of the example 12-ketolithocholic acid on colon histopathological lesions of the mice;
FIG. 6 is a comparison of the histopathological scores of groups of mice in the effect of the ketolithocholic acid of this example on colon tissue pathological lesions of colon mice;
FIG. 7 is a schematic representation of a flow scattergram of the colon ILC3s of each group of mice in this example 12-ketolithocholic acid affecting the number of colon ILC3s of mice with colon inflammation;
FIG. 8 is a graph showing comparison of the number of colon ILC3s in each group of mice in the effect of the example 12-ketolithocholic acid on the number of colon ILC3s in mice with colon inflammation;
FIG. 9 shows colon IL-17A of the mice having a colon inflammatory disease with 12-ketolithocholic acid of this example + In the influence of ILC3s number, mice of each group had colon IL-17A + ILC3s flow scatter diagram schematic;
FIG. 10 shows colon IL-17A of the mice having a colon inflammatory disease with 12-ketolithocholic acid of this example + In the influence of ILC3s number, mice of each group had colon IL-17A + ILC3s number comparison schematic.
Detailed Description
Embodiments of the present application will be described in detail below with reference to the following examples, which are to be construed as merely illustrative and not limitative of the scope of the application, but are not intended to limit the scope of the application to the specific conditions set forth in the examples, either as conventional or manufacturer-suggested, nor are reagents or apparatus employed to identify manufacturers as conventional products available for commercial purchase.
Examples:
the technical scheme of the application is based on: the 12-ketolithocholic acid can obviously reduce the symptoms of weight loss, disease activity index, colon shortening, tissue pathological damage and the like of a mouse with the ulcerative colitis model, so that the 12-ketolithocholic acid can be used for preventing and treating the ulcerative colitis.
12-Keto-lithocholic acid of the application, molecular formula C 24 H 38 O 4 Molecular weight: 390.56, the structural formula is as follows:
the 12-ketolithocholic acid is purchased from Sigma-Aldrich company, the purity is more than or equal to 95%, and the purity meets the medication standard.
1. Experimental materials
1.1 medicinal materials
12-Ketone cholic acid (CAS: 5130-29-0, HPLC. Gtoreq.95% brand: sigma-Aldrich), deoxycholic acid (CAS: 83-44-3, HPLC. Gtoreq.99% brand: sigma-Aldrich), dextran sodium sulfate (DSS, CAS number: 9011-18-1:36-50 kDa)
1.2 laboratory animals
Male mice weighing 20-22g were fed in a conventional environment and fed free-drinking water, supplied by St Bei Fu (Beijing) Biotechnology Co., ltd. Animal license number: SCXK (Beijing) 2019-0010.
2. Experimental method
2.1 establishment of ulcerative colitis model
C57BL/6 male mice of 6-8 weeks of age were selected and randomly grouped. The mice were allowed to eat free diet at the beginning of the study using the international modeling method for ulcerative colitis, and were free to drink 2.5% DSS in water for 7 days after the initial adaptation phase to induce ulcerative colitis. After successful modeling, the 1% DSS aqueous solution was continued for 2 days to maintain the colitis model state.
2.2 grouping and administration modes
(1) Control group: 6, free drinking water diet, and continuously pouring sterile physiological saline for stomach 1 time a day on the 1 st day of molding.
(2) Model group: 7, 3% DSS+1% DSS aqueous solution, free diet, and continuous gastric lavage of sterile physiological saline was started on day 1 of molding, 1 time a day.
(3) 12-ketolithocholic acid group (12-ketolithocholic acid, 12-KLCA): 6, 3% DSS+1% DSS aqueous solution and free diet were freely drunk, and 12-ketolithocholic acid aqueous solution with 2mmol/L gastric concentration was continuously infused 1 time a day starting on the 1 st day of molding.
(4) Deoxycholic acid group (DCA): 6, 3% DSS+1% DSS aqueous solution and free diet are freely drunk, and deoxycholic acid aqueous solution with 2mmol/L stomach concentration is continuously infused on the 1 st day of molding, 1 time a day.
(5) 12-Keto-lithocholic acid and deoxycholic acid combination (12-KLCA+DCA): 6, 3% DSS+1% DSS aqueous solution, free diet, and two bile acid mixture aqueous solutions with 2mmol/L concentration were continuously infused 1 time a day from the 1 st day of molding.
During the experiment, dosage formulations were freshly prepared daily, fully dissolved with deionized water, 12-ketolithocholic acid and deoxycholic acid, and gently swirled in an ultrasonic water bath to accelerate dissolution. The 3% dss solution was prepared with deionized water. All formulations were used within 1 hour of preparation.
2.3 model evaluation of ulcerative colitis
Record stool, diarrhea and weight changes daily during the experiment, after the end of the experiment, kill mice after anesthesia, open the abdominal cavity, remove the colon, measure the length of the colon, and take the proximal colon as H&E staining was used for colon histopathological observation. At the same time, analyzing colon innate immune cells ILC3s and cell subgroup IL-17A thereof + The amount of ILC3s was used to test the ability of ILC3s to secrete IL-17A and assess the ability of ulcerative colitis mice to develop a type 3 immune response in the colon.
2.4 statistical analysis
All experimental data are repeated three times, the result is expressed as mean value +/-standard deviation, SPSS statistical software is adopted to carry out single-factor analysis of variance on the experimental data, and P is less than or equal to 0.05, so that the experimental data have significant difference.
(1) The product using object is as follows: ulcerative colitis model mouse
(2) Optimal use concentration of the product: each mouse had 2mmol/L per time.
(3) The using method comprises the following steps: preparing 12-ketolithocholic acid aqueous solution with concentration of 2mmol/L, and taking orally for 1 time a day for 9 days.
3. Conclusion of the experiment
3.1 Effect of 12-ketolithocholic acid on weight loss in ulcerative colitis mice
As shown in FIG. 1, the weight change of each group of mice is shown schematically. Wherein, 12-KLCA: 12-ketolithocholic acid; DCA: deoxycholic acid; 12-KLCA+DCA: 12-ketolithocholic acid and deoxycholic acid; p <0.001 compared to control group; # P <0.001, # P <0.01 compared to model group; ΔP <0.01 compared to group 12-KLCA.
The data in fig. 1 show that DSS mice began to continuously decrease in body weight (P < 0.001) at day 6 of modeling, as compared to control. Compared with the model group, the body weight of the mice in the 12-KLCA group, the DCA group and the 12-KLCA+DCA group is recovered at the beginning of the 6 th day and tends to be stable (P < 0.001), wherein the effect of the 12-KLCA is obviously better than that of the DCA (P < 0.01), and the superiority of the 12-KLCA in the aspect of obviously improving the body weight loss caused by ulcerative colitis is reflected.
3.2 Effect of 12-ketolithocholic acid on the disease Activity index of ulcerative colitis mice
As shown in FIG. 2, the disease activity index change of each group of mice is shown in comparison. Wherein, 12-KLCA: 12-ketolithocholic acid; DCA: deoxycholic acid; 12-KLCA+DCA: 12-ketolithocholic acid and deoxycholic acid; p <0.001 compared to control group; # P <0.001, # P <0.01 compared to model group; ΔP <0.05 compared to group 12-KLCA.
The data in fig. 2 shows that model mice developed diarrhea, watery feces and even fecal occult blood from day 1 after molding to the end of the experiment, and that the disease activity index increased significantly (P < 0.001) compared to the control group. While the 12-KLCA group and the 12-KLCA+DCA group had a clear improvement from day 3 of modeling, increased appetite, gradually increased activity, gradually normal stool, and a significantly lower disease activity score than the model group (P < 0.001), while the DCA group had a significantly lower disease activity index from day 5 (P < 0.01), but the effect of 12-KLCA was significantly better than DCA (P < 0.05).
3.3 Effect of 12-ketolithocholic acid on ulcerative colitis mouse colon length
As shown in fig. 3, a schematic of colon length measurement for each group of mice is shown.
As shown in fig. 4, colon length comparison is schematically shown for each group of mice.
Wherein, 12-KLCA: 12-ketolithocholic acid; DCA: deoxycholic acid; 12-KLCA+DCA: 12-ketolithocholic acid and deoxycholic acid; p <0.01 compared to control group; compared to model group, #p <0.05; ΔP <0.05 compared to group 12-KLCA.
As can be seen from the measurement data in figure 3,
fig. 4, in combination with the analysis of the measurement data of fig. 3 above, shows that the colon of the model group mice is significantly shortened compared to the control group, whereas the 12-KLCA alone treatment significantly inhibited colonic shortening in colonic mice (P < 0.05) while the DCA combination 12-klca+dca group showed no significant change in colonic length (P > 0.05).
3.4 Effect of 12-ketolithocholic acid on colon tissue pathological lesions of colon inflammatory mice
As shown in fig. 5, microscopic diagrams of histopathology of each group of mice at 10×10 magnification are shown;
as shown in fig. 6, a comparative model of histopathological scores for each group of mice is shown.
Wherein, 12-KLCA: 12-ketolithocholic acid; DCA: deoxycholic acid; 12-KLCA+DCA: 12-ketolithocholic acid and deoxycholic acid; p <0.05 compared to control group; compared to model group, #p <0.05; ΔP <0.01, ΔP <0.05 compared to group 12-KLCA.
Fig. 5, in combination with the data of fig. 6, shows that colon tissue of mice in the model group is typical inflammatory mucosal manifestation, and that mucosal massive inflammatory cell infiltration, crypt loss, tissue structure breakage, edema and the like are seen, and that histopathological scores are significantly higher than those of the control group (P < 0.05). DCA groups and 12-klca+dca groups gave similar results to the model group. In contrast, the colon tissue of the mice in the 12-KLCA group showed reduced lesions, healed ulcers and restored crypt, and the histopathological score was significantly reduced (P < 0.05) compared to the model group, DCA group and 12-KLCA+DCA group, indicating that 12-KLCA can significantly alleviate the colon tissue pathological lesions of ulcerative colitis mice.
3.5 The effect of 12-ketolithocholic acid on the number of colon innate immune cells ILC3s in colon inflammatory mice is shown in FIG. 7, which is a schematic of a flow scattergram of colon ILC3s cells in each group of mice;
as shown in FIG. 8, a comparison of the numbers of colon ILC3s cells in each group of mice is shown.
Wherein, 12-KLCA: 12-ketolithocholic acid; DCA: deoxycholic acid; 12-KLCA+DCA: 12-ketolithocholic acid and deoxycholic acid; ILC3s, type 3innate lymphoid cells; p <0.01 compared to control group; compared to model group, #p <0.05; in contrast to the group of 12-KLCAs, deltaDeltaP <0.001, Δp <0.05.
Fig. 7 in combination with the data of fig. 8 shows that the number of colon ILC3s was significantly higher in model mice than in control mice (P < 0.01). DCA and 12-klca+dca groups were similar to the model group results. Whereas the number of mice colon ILC3s was significantly reduced (P < 0.05) in the 12-KLCA group compared to the model, DCA and 12-klca+dca groups, indicating that 12-KLCA can significantly inhibit the production of ulcerative colitis mice colon innate immune cells ILC3 s.
3.6 12-Keto-lithocholic acid colon IL-17A of colon enteritis mice + Influence of ILC3s subgroup
As shown in FIG. 9, colon IL-17A was obtained for each group of mice + ILC3s cell flow scattergram schematic;
as shown in FIG. 10, colon IL-17A was obtained for each group of mice + ILC3s cell number comparison schematic.
Wherein, 12-KLCA: 12-ketolithocholic acid; DCA: deoxycholic acid; 12-KLCA+DCA: 12-ketolithocholic acid and deoxycholic acid; ILC3s, type 3innate lymphoid cells; p <0.05 compared to control group; compared to model group, #p <0.05; ΔP <0.01, ΔP <0.05 compared to group 12-KLCA.
FIG. 9 in combination with the data of FIG. 10 shows colon IL-17A in model group mice + The number of ILC3s was significantly higher than in the control group (P<0.01)。12-KThe LCA+DCA group has similar results to the model group, while the DCA group IL-17A + The number of ILC3s is significantly higher than in model group (P<0.01)(P<0.05). Compared with the model group, DCA group and 12-KLCA+DCA group, the 12-KLCA group mice colon IL-17A + The number of ILC3s was significantly reduced (P<0.05 It is demonstrated that 12-KLCA can regulate the intestinal tract type 3 immune response by inhibiting the secretion of a pro-inflammatory factor IL-17A by the colon innate immune cells ILC3s of ulcerative colitis mice, thereby alleviating colon inflammation.
In conclusion, the 12-ketolithocholic acid can obviously reduce the weight loss and disease activity score of ulcerative colitis mice, restore the colon length, improve the pathological damage symptoms of intestinal tissue inflammation, reduce the ILC3s level of colon innate immune cells and inhibit the cells from secreting the proinflammatory cytokine IL-17A, thereby playing the role of preventing and treating exacerbation of ulcerative colitis.
Finally, it should be noted that: the foregoing description is only of the preferred embodiments of the application and is not intended to limit the scope of the application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the protection scope of the present application.
Claims (8)
- Application of 12-ketolithocholic acid in preparing medicine for preventing and treating ulcerative colitis is provided.
- 2. The use of 12-ketolithocholic acid for the preparation of a medicament for the prevention and treatment of ulcerative colitis according to claim 1, characterized in that it is useful for increasing the weight of ulcerative colitis mice, decreasing the index of disease activity, restoring colon length, alleviating damage to histopathological structures, regulating the number and function of the type 3 indigenous lymphoid cells of the colon.
- 3. Use of 12-ketolithocholic acid according to claim 2 for the preparation of a medicament for the prevention and treatment of ulcerative colitis, wherein the disease activity index comprises the rate of weight loss, diarrhea status and hematochezia status of ulcerative colitis mice.
- 4. A medicament for preventing and treating ulcerative colitis, which is characterized by taking 12-ketolithocholic acid as an active ingredient.
- 5. The medicine for preventing and treating ulcerative colitis according to claim 4, wherein the medicine further comprises pharmaceutically acceptable auxiliary materials, and is prepared into a pharmaceutically corresponding dosage form.
- 6. The drug for preventing and treating ulcerative colitis according to claim 4, wherein the HPLC purity of 12-ketolithocholic acid is not less than 95%.
- 7. The medicament for preventing and treating ulcerative colitis according to claim 5, wherein the adjuvant is a liquid, solid or semi-solid adjuvant.
- 8. The medicament for preventing and treating ulcerative colitis according to claim 5, wherein the dosage form comprises any one of a liquid dosage form, a solid dosage form and a semisolid dosage form;the liquid dosage forms include solutions and/or injections; the solid dosage forms include powders and/or tablets; the semi-solid dosage forms include suppositories and/or pastes.
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