阅读说明：本技术 选择性降低肠道次级胆汁酸生成的产品及应用 (Product for selectively reducing intestinal secondary bile acid generation and application ) 是由 沈剑萍 于 2020-04-30 设计创作，主要内容包括：本发明提供一种选择性降低肠道次级胆汁酸生成的产品及应用,所述产品含有口服定位释放制剂,所述口服定位释放制剂于结肠定位释放,所述口服定位释放制剂中含有肠道菌7α-脱羟反应抑制剂。本产品能够在结肠定位缓释释放,为多种疾病的治疗提供了技术支持。(The invention provides a product for selectively reducing generation of intestinal secondary bile acid and application thereof, wherein the product contains an oral localized release preparation which is released in a colon in a localized manner, and the oral localized release preparation contains an intestinal bacteria 7 alpha-dehydroxy reaction inhibitor. The product can be released in colon in a positioning and slow-release manner, and provides technical support for the treatment of various diseases.)

Use of a 7 α -dehydroxy reaction inhibitor in the manufacture of a colon targeted release product for reducing the production of secondary bile acids in the colon, wherein the 7 α -dehydroxy reaction inhibitor is used as an effective ingredient for reducing the production of secondary bile acids in the colon.

2. The use according to claim 1, wherein the 7 α -dehydroxylation inhibitor comprises acids which lower the pH of the environment in the colon as the main active ingredient.

3. The use according to claim 2, wherein the acid is selected from the group consisting of glutamic acid, aspartic acid, citric acid, malic acid, gluconic acid, lactic acid, aminosalicylic acid, ascorbic acid, acetic acid, propionic acid, butyric acid, fatty acid, or a combination of any one or more of their pharmaceutically acceptable salts.

4. A product for reducing the production of secondary bile acids in the colon, said product comprising an oral targeted release formulation for targeted release in the colon, said oral targeted release formulation comprising a 7 α -dehydroxy inhibitor.

5. The product according to claim 4, wherein the 7 α -dehydroxylation inhibitor comprises acids capable of lowering the pH of the environment in the colon as the main active ingredient.

6. The product according to claim 5, wherein the acid is selected from the group consisting of glutamic acid, aspartic acid, citric acid, malic acid, gluconic acid, lactic acid, aminosalicylic acid, ascorbic acid, acetic acid, propionic acid, butyric acid, fatty acid, or a combination of any one or more thereof in a pharmaceutically acceptable salt.

7. The product of claim 4, wherein the oral localized release formulation is a time-lag coating, a pH sensitive enteric coating, or a coating of a polymeric material that is activatable by colonic flora enzyme systems.

8. The product of claim 4, wherein the 7 α -dehydroxy inhibitor is supported in a sustained release carrier, and the sustained release carrier and the 7 α -dehydroxy inhibitor are encapsulated in an oral localized release formulation.

9. The product of claim 8, wherein the sustained release carrier is selected from the group consisting of: any one of resin, activated carbon, ethyl cellulose, or silica.

Use of an alpha-dehydroxy reaction inhibitor for the preparation of a colon-targeted release medicament for the treatment of non-alcoholic fatty liver disease, diabetes, obesity, liver cancer, diabetes, colorectal cancer, colorectal polyps, or ulcerative colitis.

11. The use according to claim 10, wherein the 7 α -dehydroxylation inhibitor comprises acids which lower the pH of the environment in the colon as the main active ingredient.

12. The use according to claim 11, wherein the acid is selected from the group consisting of glutamic acid, aspartic acid, citric acid, malic acid, gluconic acid, lactic acid, aminosalicylic acid, ascorbic acid, acetic acid, propionic acid, butyric acid, fatty acid, or a combination of any one or more thereof in a pharmaceutically acceptable salt.

13. The use according to claim 10, wherein the oral localized release formulation is a time lag effect coating, a pH sensitive enteric coating or a coating of a polymeric material that can be activated by colonic flora enzyme systems.

14. The use according to claim 10, wherein the 7 α -dehydroxy reaction inhibitor is supported in a slow release carrier, and the slow release carrier and the 7 α -dehydroxy reaction inhibitor are encapsulated in an oral targeted release formulation.

15. Use according to claim 14, wherein the slow release carrier is selected from the group consisting of: any one of resin, activated carbon, ethyl cellulose, or silica.

16. A medicine for treating nonalcoholic fatty liver disease, diabetes, obesity, liver cancer, diabetes, colorectal cancer, colorectal polyp or ulcerative colitis is characterized in that the main active ingredient of the medicine is a 7 alpha-dehydroxy reaction inhibitor capable of being released in a colon-specific manner.

17. The medicament according to claim 16, wherein the main active ingredient of the 7 α -dehydroxy reaction inhibitor is an acid capable of lowering the pH of the environment in the colon.

18. The medicament of claim 17, wherein the acid is selected from the group consisting of glutamic acid, aspartic acid, citric acid, malic acid, gluconic acid, lactic acid, aminosalicylic acid, ascorbic acid, acetic acid, propionic acid, butyric acid, fatty acid, and a combination of any one or more thereof in a pharmaceutically acceptable salt.

19. The medicament of claim 16, wherein the oral targeted release formulation is a time-lag coating, a pH sensitive enteric coating, or a coating of a polymeric material that is activatable by colonic flora enzyme systems.

20. The medicament of claim 16, wherein the 7 α -dehydroxy inhibitor is supported in a sustained release carrier, and the sustained release carrier and the 7 α -dehydroxy inhibitor are encapsulated in an oral targeted release formulation.

21. The medicament of claim 20, wherein the sustained release carrier is selected from the group consisting of: any one of resin, activated carbon, ethyl cellulose, or silica.

Technical Field

The invention belongs to the field of drug research and development, and particularly relates to a composition capable of selectively inhibiting intestinal secondary bile acid and application of the composition in treating non-alcoholic fatty liver disease, liver cancer, diabetes, metabolic syndrome, colorectal cancer, colorectal polyp, ulcerative colitis and other diseases.

Background

Bile acids are produced in the liver by cholesterol conversion. Bile acid is the main component in bile, and accounts for about 85% of the solid components in bile. Bile acids are natural ionized detergents that play an important role in fatty acid absorption, transport, secretion, and regulation of cholesterol metabolism. Bile acids present in human bile are mainly: cholic acid, chenodeoxycholic acid, deoxycholic acid, and a small amount of lithocholic acid. Under normal conditions, the liver of human beings has low bile acid synthesis speed, and the liver of human beings synthesizes about 400-600 mg per day. Enterohepatic circulation of bile acids refers to the process in which, after secretion of bile acids into the intestinal lumen, in the terminal jejunum or ileum, and back again into the liver via the portal vein system, about 95% of the bile acids are reabsorbed back into the liver and only 5% of the bile acids are excreted through the feces. The normal bile acid pool in human liver is about 3-5 g, while the organism maintains the digestion and absorption of lipid substances, about 12-32 g of bile acid is needed, and the intestinal liver circulation for about 12 times after meal every day can make up the synthesis deficiency of the bile acid in liver, so that the limited bile acid pool can play the role of emulsification to the maximum extent. Therefore, the physiological significance of the enterohepatic circulation of bile acids is to allow limited reuse of bile acids and to promote the normal digestion and absorption of lipid foods.

The liver is the only organ that synthesizes bile acids, which are synthesized by cholesterol in the parenchymal hepatocytes, producing primary bile acids in the free state: chenodeoxycholic acid and cholic acid are combined with taurine/glycine or sulfate/glucuronic acid by amido bond to form completely ionized hydrophilic polar molecule with negative charge, namely combined bile acid (tauro/glycocholic acid, T/G-BA; sulfate/glucuronic acid cholic acid, S/U-BA). The conjugated bile acid is secreted into the intestinal tract by the bile duct, has extremely high solubility in the pH environment of the intestinal tract, and is beneficial to exerting physiological action.

In the intestine, bound bile acids dissociate into free bile acids under the action of intestinal bacteria at the terminal ileum and upper colon, and are subsequently converted into secondary bile acids by 7 α -dehydroxylation of intestinal bacteria in the colon: deoxycholic acid and lithocholic acid, both reabsorbed by diffusion in the colon or excreted by the feces.

The conjugated bile acid can hardly be absorbed at the front end of small intestine, but most of T/G-BA can be actively and effectively reabsorbed into intestinal epithelial cells by apical sodium-dependent bile acid transporter [ ASBT ] of apical membrane in ileum, reabsorbed into portal vein and transported back to liver with blood flow, and the reabsorbed bile acid is secreted into biliary system by bile salt output pump [ BESP (ABCB11) ] of liver cell capillary biliary tract membrane together with newly synthesized conjugated bile acid in liver cell. And the S/U-BA is reabsorbed into the intestinal epithelial cells by the multidrug resistance related protein 2[ MRP2(ABCC2) ] of the apical membrane of the intestinal epithelial cells, is pumped into the portal vein system by MRP3 of the basement membrane, and is secreted into the capillary bile duct by MRP2 of the liver cell capillary bile duct membrane.

As shown in fig. 8, the enterosymbiotic bacteria convert primary bile acid generated by the host into secondary bile acid through 3 main steps, firstly deaminate the primary bile acid to generate unbound bile acid, secondly transport the unbound bile acid into symbiotic bacteria with bile acid metabolism ability, and thirdly dehydroxylate the unbound bile acid by 7-alpha to finally generate final secondary bile acid (deoxycholic acid and lithocholic acid).

Bile acid has important physiological functions and certain cytotoxicity. The cytotoxicity of different bile acids varies, and in general, the more hydrophobic the bile acid becomes, the more hydrophobic the secondary bile acid produced by the metabolism of the intestinal flora is than the primary bile acid, and as early as 1940, deoxycholic acid (DCA) in the secondary bile acid has been shown to be carcinogenic. Some patients with liver disease present with a disturbed intestinal flora, wherein the flora associated with bile acid metabolism such as Bacteroides and Clostridium is significantly elevated, producing large amounts of secondary bile acids causing damage to enterocytes and hepatocytes. In recent years, more and more researches prove that secondary bile acid can cause toxicity to host cells, stimulation to intestinal cells can cause inflammation, hyperplasia and intestinal cancer of the intestinal cells, and entering liver through the circulation of liver and intestine can cause inflammation, fibrosis and even canceration of liver cells.

Non-alcoholic fatty liver disease comprises a series of diseases, from abnormal deposition of simple lipids in the liver (simple steatosis), to steatosis accompanied by inflammation to non-alcoholic steatohepatitis (NASH) and even cirrhosis, resulting in liver canceration, without a history of alcoholism. Fatty liver and non-alcoholic steatohepatitis can develop in various age groups including children. In the United states, 10-20% of the population suffers from fatty liver, and 3-5% suffer from non-alcoholic steatohepatitis. It is estimated that the incidence of nonalcoholic fatty liver disease in more developed cities in china is about 2-fold higher in 20 years. The incidence of fatty liver or simple fatty changes in the population is 10-15%, and in obese people is as high as 70-80%. The incidence rate of the non-alcoholic fatty liver disease in common people is about 3 percent, and the incidence rate in obese people can reach 15 to 20 percent. Non-alcoholic steatohepatitis is one of the typical manifestations of metabolic syndrome. The metabolic syndrome is characterized by central obesity, insulin resistance, hypertension, hyperglycemia, hypertriglyceridemia and fatty liver. Currently, there is no accepted standard treatment regimen for non-alcoholic steatoliver disease and non-alcoholic steatohepatitis. The primary conditions for the treatment of non-alcoholic steatoliver disease and non-alcoholic steatohepatitis are to change lifestyle, strengthen exercise, avoid drinking, reduce body weight, balance nutrition and diet. Although the efficacy is not very positive, there are mechanisms to apply some antidotes such as hepatocyte protection drugs, etc. The promising drugs include antioxidants, insulin sensitizers (e.g., metformin and thiazolidinediones) hepatoprotectants and lipid lowering drugs. However, the use of these drugs is controversial and more research is required to confirm their efficacy and safety. There is a need to develop new safe methods of preventing and/or reversing the accumulation of fat in the liver.

Colorectal cancer is a common malignancy, listed at position 3 and 2 in male and female malignancies, respectively, accounting for approximately 8% of cancer deaths worldwide. The incidence of colorectal cancer is increasing year by year. The development and clinical features of colorectal cancer depends largely on the precautions taken, and recent statistics of cancer in the united states indicate a timely basis. The endoscope detection is obtained, and the precancerous lesions are removed, and the incidence rate of the precancerous lesions tends to decrease. Colorectal polyps are the most common disease in clinic, most of which are adenomatous polyps, while 70% of patients with colorectal cancer develop from the progression of colorectal adenomatous polyps. Colorectal adenoma is the most main precancerous lesion of colorectal cancer, the currently recognized polyp development modes at home and abroad are adenomatous polyp, dysplasia and adenocarcinoma, and the canceration process lasts about 10 years, so the treatment of colorectal polyp is particularly important.

In recent years, the incidence of colorectal cancer is increasing with westernization of people's dietary habits. High fat diet is one of the important factors for promoting colorectal cancer development, and can increase the level of secondary bile acid in intestinal cavity. Secondary bile acids are closely related to the occurrence of colorectal cancer, especially deoxycholic acid, which is considered to be a carcinogenic and carcinogenic factor.

At present, some medicines aim at the pathway of bile acid liver-intestine circulation and are used for treating diseases such as non-alcoholic fatty liver disease (NASH), diabetes, obesity and the like, for example, ASBT receptor inhibitors developed by foreign pharmaceutical companies can inhibit bile acid reabsorption in intestinal tracts, are expected to be used for treating NASH, but are stopped due to serious side effects; cholestyramine can absorb bile acid in intestinal tracts, is used for reducing blood cholesterol, and is also rarely used clinically due to common side effects and the like. In addition, cholestyramine has no therapeutic effect on nonalcoholic fatty liver disease (NASH), diabetes, colon cancer, and the like.

It can be seen that the treatment method for inhibiting all bile acid hepatic-intestinal circulation without targeting and selectivity can seriously interfere with the normal physiological functions of bile acid, cause damage to the organism and have poor curative effect. As described above, 95% of bile acids are reabsorbed before entering the colon, and only 5% of bile acids enter the colon and are metabolized by the intestinal flora to finally form secondary bile acids (deoxycholic acid, lithocholic acid, etc.), which are closely related to body diseases.

Research shows that the 7 alpha-dehydroxylation reaction of intestinal bacteria is sensitive to the pH of the environment, the optimal enzyme activity pH is about 7, the pH is reduced to 6, the enzyme activity is only 25 percent and is reduced to below 5, and the enzyme is almost inactivated.

There is currently no drug that selectively inhibits secondary bile acids in the colon.

Disclosure of Invention

In view of the above-mentioned disadvantages of the prior art, the present invention aims to provide a drug for selectively inhibiting intestinal secondary bile acid and its application, which fills the gap in the prior art.

To achieve the above and other related objects, the present invention provides the use of a 7 α -dehydroxy reaction inhibitor for the preparation of a colon targeted release product for reducing the production of secondary bile acids in the colon, wherein the 7 α -dehydroxy reaction inhibitor is an effective ingredient for reducing the production of secondary bile acids in the colon.

That is, the 7 α -dehydroxylation inhibitor is the only active ingredient or one of the active ingredients that reduces the production of secondary bile acids in the colon.

Furthermore, the main effective component of the 7 alpha-dehydroxy reaction inhibitor is an acid capable of reducing the pH value of the environment in the colon.

Further, the acid is selected from the group consisting of glutamic acid, aspartic acid, citric acid, malic acid, gluconic acid, lactic acid, aminosalicylic acid, ascorbic acid, acetic acid, propionic acid, butyric acid, fatty acid, or a combination of any one or more thereof of pharmaceutically acceptable salts thereof.

Further, the common techniques for colon targeted drug delivery systems are: a. coating by using time lag effect; a pH sensitive enteric coating; coating with a polymeric material which is activatable by colonic flora enzyme systems. The macromolecular material coating which can be activated by colonic flora enzyme system utilizes the unique enzyme system activation coating of colonic flora, a plurality of macromolecular materials are degraded by the enzymes in colon, and the macromolecular materials as drug carriers can not be degraded in stomach and small intestine due to the lack of corresponding enzymes, so that the drug can not be released in stomach and small intestine, for example, pectin, guar gum, azo polymers and alpha, beta, gamma-cyclodextrin can be used as the carrier material of colonic drug delivery system. The main innovation of the present invention is the concept of the whole invention, and those skilled in the art can select suitable coatings according to the prior art means to achieve colon specific release of 7 α -dehydroxy inhibitor, and the coatings capable of achieving colon specific release are all in the same spirit of the present invention, therefore, the present application is not limited to the above three specific materials or types.

Further, the 7 alpha-dehydroxy reaction inhibitor is loaded in a slow release carrier, and the slow release carrier and the 7 alpha-dehydroxy reaction inhibitor are wrapped in an oral positioning release preparation.

The slow release function is realized by adopting a slow release carrier in the prior art. Still further, the sustained release carrier is selected from the group consisting of: any one of resin, activated carbon, ethyl cellulose, or silica.

The slow release carrier can slowly and uniformly release the 7 alpha-dehydroxy reaction inhibitor in the colon for several hours, thereby persistently reducing the pH in the colon. Preferably, the slow release carrier has the function of adsorbing bile acid in the intestinal tract. The slow release carrier with the function can also absorb a part of bile acid in intestinal tracts.

Further, the present invention provides a product for reducing secondary bile acid production in the colon, said product comprising an oral targeted release formulation for targeted release in the colon, said oral targeted release formulation comprising a 7 α -dehydroxy response inhibitor.

The product has colon positioning and releasing functions, and can be positioned to achieve colon release.

The intestinal symbiotic bacteria convert primary bile acid generated by a host into secondary bile acid through 3 main steps, firstly, the primary bile acid is combined to be deaminated to generate unconjugated bile acid, secondly, the unconjugated bile acid is transported into symbiotic bacteria with bile acid metabolic capability, and thirdly, the unconjugated bile acid is subjected to 7-alpha dehydroxylation reaction to finally generate final secondary bile acid (deoxycholic acid and lithocholic acid).

The 7 α -dehydroxylation inhibitor herein refers to a substance that inhibits the 7 α -dehydroxylation in the third step. That is, the 7 α -dehydroxylation inhibitor is a substance that inhibits the 7 α -dehydroxylation of enteric bacteria. The 7 alpha-dehydroxylation inhibitor can inhibit the 7 alpha-dehydroxylation of intestinal bacteria by enhancing the acidity of the environment in the colon, so as to inhibit the reaction efficiency of unconjugated bile acid to generate final secondary bile acid through 7-alpha dehydroxylation, and finally achieve the purpose of inhibiting the generation of the secondary bile acid.

Furthermore, the main effective component of the 7 alpha-dehydroxy reaction inhibitor is an acid capable of reducing the pH value of the environment in the colon.

More preferably, the acid is selected from the group consisting of glutamic acid, aspartic acid, citric acid, malic acid, gluconic acid, lactic acid, aminosalicylic acid, ascorbic acid, acetic acid, propionic acid, butyric acid, fatty acid, or a combination of any one or more of their pharmaceutically acceptable salts.

Further, the oral targeted release formulation is state of the art. Common techniques for oral colon-specific drug delivery systems are: a. coating by using time lag effect; a pH sensitive enteric coating; c. coating with polymer material capable of being activated by colonic flora enzyme system. The macromolecular material coating which can be activated by colonic flora enzyme system utilizes the unique enzyme system activation coating of colonic flora, a plurality of macromolecular materials are degraded by the enzymes in colon, and the macromolecular materials as drug carriers can not be degraded in stomach and small intestine due to the lack of corresponding enzymes, so that the drug can not be released in stomach and small intestine, for example, pectin, guar gum, azo polymers and alpha, beta, gamma-cyclodextrin can be used as the carrier material of colonic drug delivery system. The main innovation of the present invention is the concept of the whole invention, and those skilled in the art can select suitable coatings according to the prior art means to achieve colon specific release of 7 α -dehydroxy inhibitor, and the coatings capable of achieving colon specific release are all in the same spirit of the present invention, therefore, the present application is not limited to the above three specific materials or types.

Further, the 7 alpha-dehydroxy reaction inhibitor is loaded in a slow release carrier, and the slow release carrier and the 7 alpha-dehydroxy reaction inhibitor are wrapped in an oral positioning release preparation.

The slow release function is realized by adopting a slow release carrier in the prior art. Still further, the sustained release carrier is selected from the group consisting of: any one of resin, activated carbon, ethyl cellulose, or silica.

The slow release carrier can slowly and uniformly release the 7 alpha-dehydroxy reaction inhibitor in the colon for several hours, thereby persistently reducing the pH in the colon. Preferably, the sustained-release carrier has a function of adsorbing bile acid in the intestinal tract after releasing the 7 α -dehydroxy reaction inhibitor. The slow release carrier with the function can also absorb a part of bile acid in intestinal tracts.

In one embodiment, the product for inhibiting secondary bile acids in the colon comprises at least 3 layers, wherein the core component is the 7 alpha-dehydroxylation inhibitor, the core component is positioned in a slow release carrier, and the outermost layer wraps a colon positioning carrier.

Different slow release carriers and colon-specific release preparations are adopted, so that the amount of the 7 alpha-dehydroxy reaction inhibitor which can be loaded is different, the mass ratio of the colon-specific release preparations to the slow release carriers is different, and the application does not limit the weight ratio. In general, one skilled in the art can obtain formulations with higher drug loading based on experimental methods in the prior art in combination with specifically employed materials.

The main innovation of the present invention is the concept of the whole invention, and those skilled in the art can select a suitable slow release carrier according to the prior art means to achieve the slow release of the 7 α -dehydroxylation inhibitor, so that the carrier which can slowly release the 7 α -dehydroxylation inhibitor is consistent with the concept of the present invention, and therefore, the present application is not limited to the specific material or type of the above slow release carrier.

The product is generally an oral preparation.

In another aspect of the invention, there is provided a use of the 7 α -dehydroxy reaction inhibitor in the preparation of a colon-targeted release medicament for the treatment of non-alcoholic fatty liver disease, diabetes, obesity, liver cancer, diabetes, colorectal cancer, colorectal polyps, or ulcerative colitis.

Furthermore, the main effective component of the 7 alpha-dehydroxylation inhibitor is an acid capable of reducing the pH value of the environment in the colon.

More preferably, the acid is selected from the group consisting of glutamic acid, aspartic acid, citric acid, malic acid, gluconic acid, lactic acid, aminosalicylic acid, ascorbic acid, acetic acid, propionic acid, butyric acid, fatty acid, or a combination of any one or more of their pharmaceutically acceptable salts.

Further, the oral targeted release formulation is state of the art. Common techniques for oral colon-specific drug delivery systems are: a. coating by using time lag effect; a pH sensitive enteric coating; c. coating with polymer material capable of being activated by colonic flora enzyme system. The macromolecular material coating which can be activated by colonic flora enzyme system can utilize the unique enzyme system activation coating of colonic flora, a plurality of macromolecular materials are degraded by the enzymes in colon, and the macromolecular materials as drug carriers can not be degraded in stomach and small intestine due to the deficiency of corresponding enzymes, so that the drug can not be released in stomach and small intestine, for example, pectin, guar gum, azo polymers and alpha, beta, gamma-cyclodextrin can be used as the carrier material of colonic drug delivery system. The person skilled in the art can select suitable coatings according to the prior art.

Further, the 7 alpha-dehydroxy reaction inhibitor is loaded in a slow release carrier, and the slow release carrier and the 7 alpha-dehydroxy reaction inhibitor are wrapped in an oral positioning release preparation.

The slow release function is realized by adopting a slow release carrier in the prior art. Still further, the sustained release carrier is selected from the group consisting of: any one of resin, activated carbon, ethyl cellulose, or silica. The person skilled in the art can load the drug into the above-mentioned carriers according to experimental methods of the prior art, reference methods such as "application of ion exchange resin drug carriers in drug delivery systems" (ion exchange and adsorption, 2010, 26 (1): 89-96); the research progress of active carbon for medicine slow release carrier [ J ] novel chemical materials 2014(05) 10-12. specifically, 7 alpha-dehydroxylation inhibitor is loaded in the slow release carrier, which can be seen in embodiments 2-4 of the invention.

The slow release carrier can slowly and uniformly release the 7 alpha-dehydroxy reaction inhibitor in the colon for several hours, thereby persistently reducing the pH value of the environment in the colon. Preferably, the slow release carrier has the function of adsorbing bile acid in the intestinal tract, and the slow release carrier with the function can also adsorb a part of bile acid in the intestinal tract.

In one embodiment, the 7 α -dehydroxylation inhibitor with colon targeting function comprises at least 3 layers, wherein the core component is the 7 α -dehydroxylation inhibitor, the core is positioned in the slow release carrier, and the outermost layer wraps the colon targeting carrier.

In another aspect of the invention, the invention provides a medicament for treating non-alcoholic fatty liver disease, diabetes, obesity, liver cancer, diabetes, colorectal cancer, colorectal polyp or ulcerative colitis, wherein the main active ingredient of the medicament is a 7 alpha-dehydroxy reaction inhibitor capable of being released in a colon-specific manner.

Furthermore, the main effective component of the 7 alpha-dehydroxy reaction inhibitor is an acid capable of reducing the pH value of the environment in the colon.

More preferably, the acid is selected from the group consisting of glutamic acid, aspartic acid, citric acid, malic acid, gluconic acid, lactic acid, aminosalicylic acid, ascorbic acid, acetic acid, propionic acid, butyric acid, fatty acid, or a combination of any one or more of their pharmaceutically acceptable salts.

Further, the oral targeted release formulation is state of the art. Common techniques for oral colon-specific drug delivery systems are: a. coating by using time lag effect; a pH sensitive enteric coating; c. coating with polymer material capable of being activated by colonic flora enzyme system. The macromolecular material coating which can be activated by colonic flora enzyme system can utilize the specific enzyme system activation coating of colonic flora, a plurality of macromolecular materials are degraded by the enzymes in colon, and the macromolecular materials as drug carriers can not be degraded in stomach and small intestine due to the lack of corresponding enzymes, thus ensuring that the drug is not released in stomach and small intestine. Such as pectin, guar gum, azo polymers and alpha, beta, gamma-cyclodextrin can be used as carrier materials for colonic delivery systems.

Further, the 7 alpha-dehydroxy reaction inhibitor is loaded in a slow release carrier, and the slow release carrier and the 7 alpha-dehydroxy reaction inhibitor are wrapped in an oral positioning release preparation.

The slow release function is realized by adopting a slow release carrier in the prior art. Still further, the sustained release carrier is selected from the group consisting of: any one of resin, activated carbon, ethyl cellulose, or silica.

The slow release carrier can slowly and uniformly release the 7 alpha-dehydroxy reaction inhibitor in the colon for several hours, thereby persistently reducing the pH value of the environment in the colon. Preferably, the slow release carrier has the function of adsorbing bile acid in the intestinal tract, and the slow release carrier with the function can also adsorb a part of bile acid in the intestinal tract.

In one embodiment, the therapeutic drug comprises at least 3 layers, wherein the core component of the therapeutic drug is the 7 alpha-dehydroxy reaction inhibitor, the core is positioned in the slow release carrier, and the outermost layer wraps the colon positioning carrier.

As described above, the drug for selectively reducing the generation of the intestinal secondary bile acid and the application thereof of the invention have the following beneficial effects:

the medicine prepared by the application can be positioned in colon for slow release, selectively reduces the generation of secondary bile acid in intestinal tract, and provides a new possibility for treating various diseases. Experiments also prove that the pharmaceutical preparation prepared by the method can effectively reduce the generation of secondary bile acid, and has positive significance for treating various diseases.

Drawings

FIG. 1 shows the effect of pH on the 7. alpha. -dehydroxylation of enteric bacteria in example 1 of the present invention

FIG. 2 shows the digestion and absorption in the small intestine and colon of the colon-specific release sustained-release glutamic acid preparation prepared in example 3 of the present invention

FIG. 3 shows the digestion and absorption of the sustained release citric acid formulation prepared in example 4 of the present invention in the small intestine and colon

FIG. 4 shows the non-alcoholic steatohepatitis and liver cancer pathological section treated by the sustained-release glutamic acid preparation in example 7 of the present invention (A, B represents a liver tissue pathological section of a 12-week established non-alcoholic steatohepatitis-liver fibrosis-liver cancer model, C represents a control group continuing for 12 weeks, D, E represents a treatment group of the sustained-release glutamic acid preparation for colon-specific release continuing for 12 weeks)

FIG. 5 shows the serum ALT index after the treatment of the sustained-release glutamic acid preparation of each experimental group in example 7 of the present invention

FIG. 6 shows that the sustained-release glutamic acid preparations of the experimental groups of example 8 of the present invention can significantly improve blood glucose metabolism, fasting blood glucose and insulin resistance after treatment

FIG. 7 shows a slice of example 9 of the present invention showing the sustained release citric acid preparation for the treatment of colorectal adenoma and carcinoma

FIG. 8 is a schematic diagram showing the process of converting primary bile acid produced by a host into secondary bile acid for enterobacteria

Detailed Description

The present invention selectively inhibits the amount of secondary bile acids in the intestinal tract without affecting primary bile acids and without affecting the gastrointestinal circulation of 95% of bile acids. In fact, a great deal of previous research and clinical medication proves that the direct interference on the liver-intestine circulation of all bile acids causes serious side effects, such as liver damage, serious constipation, influence on the absorption of fat-soluble vitamins and other nutrient components, bleeding tendency, gallstones and the like, and the curative effect is not good enough. The present invention solves these problems.

The 7 α -dehydroxylation by intestinal bacteria is the last step in the conversion of bile acids to primary bile acids, and often the exclusive anaerobes and gram-negative bacteria in the intestine express enzymes with 7 α -dehydroxylation. Other scientific researches show that the gram-negative bacteria expressing 7 alpha-dehydroxylation reaction in intestinal tracts of patients with certain diseases such as diabetes, steatohepatitis, intestinal polyps and the like are obviously increased. Intestinal bacteria have a wide variety of 7 α -dehydroxylation reactions, and it is therefore critical to develop a molecule that can inhibit these 7 α -dehydroxylation reactions in a broad spectrum. The research of the applicant shows that the 7 alpha-dehydroxylation reaction of the intestinal bacteria has a commonality that the intestinal bacteria are sensitive to the pH of the environment, the optimum enzyme activity pH is about 7, the pH is reduced to 6, the enzyme activity is only 50 percent and is reduced below 5, and the enzyme activity is less than 25 percent.

We screened a large number of molecules that could be used to lower intestinal pH, ideally some more acidic edible and medicinal organic acids, short chain fatty acids, or acidic amino acids. However, these molecules are rapidly degraded and absorbed in the upper digestive tract after oral administration and hardly reach the colon.

The molecules inhibiting the 7 alpha-dehydroxy reaction are coated in a colon-specific release system, so that the drug molecules in the coating can be protected from degradation or absorption after oral administration and can not be released until reaching the colon.

There are a number of formulations currently available in the art for delivering substances to the colon for release. The oral colon-specific release preparation can keep the integrity in the stomach and jejunum after being taken orally, and can release substances according to the design requirement after entering the tail end of the ileum, thereby achieving the purposes of quick release and slow release. For example, enteric coated release formulations may be employed. That is, a polymer soluble in an appropriate pH range may be selected as required. Timed release formulations may also be used, the timing and location of the drug release being controlled by varying the amount of time lag between release of the formulation. The coating can also be degraded by using enzymes unique to colonic flora such as colonic beta-glucosidase to activate the release of the drug therein, and many high molecular materials are degraded by the enzymes in the colon, and the high molecular materials can not be degraded in the stomach and the small intestine due to the lack of the corresponding enzymes as drug carriers, so that the drug is not released in the stomach and the small intestine. Such as pectin, guar gum, azo polymers and alpha, beta, gamma-cyclodextrin can be used as carrier materials for colonic delivery systems.

Examples of oral terminal ileal site-directed release formulations include, but are not limited to, the methods disclosed in Review articles or patents, such as Colon Targeted Drug Delivery Systems A Review on Primary and Novel applications Oman Medical Journal 2010, Volume 25, Issue 2; design Trends and applications, AAPS PharmSciTech, Vol.16, No.4, August 2015; and cn201680001612. x; US9993435B 2; the technical scheme is described in US 5525634, US5866619, US 9993435. Such dosage forms may use hydroxypropylmethylcellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, or combinations thereof in varying proportions to provide the desired release profile to provide colonic release of one or more active ingredients.

Preferably, we use a colon-specific enzyme system activation system (or in combination with a pH sensitive coating) to deliver molecules that inhibit the 7 α -dehydroxylation reaction to the colon. The colon-specific enzyme system activation layer mainly comprises polysaccharide substances such as pectin and the like, and is coated outside active ingredients (molecules for preparing 7 alpha-dehydroxylation reaction); the pH sensitive layer adopts any existing polymer material composition which can be dissolved under the condition that the pH reaches or exceeds 7, and is coated outside the unique enzyme system activation layer of the colon.

Another important design of the present invention is to load the active ingredient in a sustained release carrier with a colon-localizing layer on the outside. The slow release carrier can avoid the explosive release of the active ingredients, so that the compound is slowly and uniformly released in the colon for several hours, and the pH value in the colon and intestinal tracts is reduced for a long time.

The slow release technology is the prior art, the currently commonly used slow release carriers comprise resin, activated carbon, ethyl cellulose, silicon dioxide and the like, and the slow release carriers can enable the compound to be slowly and uniformly released in a colon tract for 6 hours, so that the pH value in the colon intestine is durably reduced, the activity of 7 alpha-dehydroxylation reaction is durably inhibited, and the concentration of secondary bile acid is reduced.

Conditions treated by ingestion of such formulations include, but are not limited to, non-alcoholic fatty liver disease, diabetes, colorectal cancer, colorectal polyps, ulcerative colitis, crohn's disease, metabolic syndrome.

The therapeutically effective amount of the pharmaceutical composition of the present invention will depend on a number of factors. For example, the race/species, age and weight of the subject, the precise condition to be treated and its severity, the nature of the formulation. The therapeutically effective amount should ultimately be at the discretion of the attendant physician. In any event, a pharmaceutically effective amount for treating a patient suffering from non-alcoholic fatty liver disease, diabetes, colorectal cancer, colorectal polyps, ulcerative colitis and related conditions will generally be from 0.1 to 5000mg/kg of recipient (mammal) body weight per day. More typically, an effective amount will be 1 to 200mg/kg body weight/day. Thus, for an adult mammal of 50kg body weight, the actual amount per day is typically from 50mg to 10 g. This amount may be administered in a single dose per day or in several (e.g. 2, 3, 4, 5 or more) divided doses per day, such that the total daily dose is the same.

The formulations of the present invention may be combined with one or more other pharmaceutically active agents in the treatment of a disease.

The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention. It is to be understood that the processing equipment or apparatus not specifically identified in the following examples is conventional in the art. Furthermore, it is to be understood that one or more method steps mentioned in the present invention does not exclude that other method steps may also be present before or after the combined steps or that other method steps may also be inserted between these explicitly mentioned steps, unless otherwise indicated; it is also to be understood that a combined connection between one or more devices/apparatus as referred to in the present application does not exclude that further devices/apparatus may be present before or after the combined device/apparatus or that further devices/apparatus may be interposed between two devices/apparatus explicitly referred to, unless otherwise indicated. Moreover, unless otherwise indicated, the numbering of the various method steps is merely a convenient tool for identifying the various method steps, and is not intended to limit the order in which the method steps are arranged or the scope of the invention in which the invention may be practiced, and changes or modifications in the relative relationship may be made without substantially changing the technical content.

Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments, and is not intended to limit the scope of the present invention; in the description and claims of the present application, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.

When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.

Example 1 Effect of pH on the Activity of intestinal bacteria 7 alpha-Dehydroxylation

Clostridium mollicum (Clostridium leptum) anaerobic culture in vitro (80% nitrogen-10% carbon dioxide-10% hydrogen) was investigated for its activity in the conversion of cholic acid into secondary bile acid deoxycholic acid by 7 α -dehydroxylation reaction in different pH environments. Clostridium tender suspension +2uM cholic acid +50mM potassium dihydrogen phosphate buffer, adding an amount of citric acid or glutamic acid to adjust pH to 6 or 5. 60 minutes at 37 degrees celsius. At the end of the reaction, 1.0ml of 0.5N hydrochloric acid was added to the tube to terminate the enzyme reaction.

The activity of 7 alpha-dehydroxylation reaction is shown in figure 1, which confirms that the optimum enzyme activity pH of 7 alpha-dehydroxylation reaction of intestinal bacteria is 6.5-7, the pH is reduced to 6, the enzyme activity is only 50%, and is reduced to below 5, and the enzyme activity is less than 20%.

EXAMPLE 2 sustained Release formulation of glutamic acid-717 resin

In this example, glutamic acid was used as an active molecule for suppressing the 7 α -dehydroxylation of intestinal bacteria and strongly basic anion exchange resin 717 (Amberlite 717, 717 for short) was used as a sustained release carrier.

Pretreatment of i.717 resin

A 717 resin copolymer of styrene-divinylbenzene was used to encapsulate glutamic acid. It has a quaternary ammonium cationic group and active chloride ions. By substituting chloride ions attached to quaternary ammonium groups, drug ions can be loaded into the resin. Specifically, a quantity of 717 resin was washed by immersion in 50 ℃ deionized water to remove water soluble impurities, and then transferred to 95% ethanol with stirring to remove organic impurities. After washing with deionized water until no residual ethanol was present, it was dried under vacuum at 50 ℃. The predried 717 resin was immersed in a 0.1mol/l hydrochloric acid solution for 24h with constant stirring, then washed to neutrality with deionized water and dried to give the anion (Cl)^-) And (4) exchanging resin.

Preparing a sustained release formulation of glutamic acid-717 resin

Ion exchange resins encapsulate drugs mainly by ion exchange reactions. In this example, we used glutamic acid. Purified glutamic acid (0.50g) was dissolved in 50mL of deionized water under magnetic stirring at 40 deg.C, 1g of resin was added, and stirring was continued for 12 h. And pouring out the supernatant, washing with water for a plurality of times, and naturally drying to obtain the glutamic acid loaded resin.

An amount of the glutamic acid-loaded resin was transferred to a small beaker, 50ml of a solution (formic acid: water 75: 25, v/v) was added, a sample was collected from the TBSS aqueous solution every predetermined time period, and the loading amount of glutamic acid was measured and calculated by High Performance Liquid Chromatography (HPLC). The drug loading (Q) of the resin glutamic acid was calculated according to the following formula:

Qt＝V/WR.(C0-Ct)

where Qt is the drug loading of the resin at time t. C₀Is the initial drug concentration, C_tIs the drug concentration at time t. V is the volume of the liquid medicine, W_RIs the mass of the resin.

EXAMPLE 3 preparation of a sustained Release Glutamine formulation with colonic localized Release

The colon-specific enzyme system activation layer mainly contains polysaccharides such as pectin and the like and is coated outside the active ingredients; the pH sensitive layer adopts any polymer material composition which can be dissolved under the condition that the pH value reaches or exceeds 7, and is coated outside the special enzyme system activation layer of the colon. The active ingredient is a molecule that inhibits 7 alpha-dehydroxylation.

20% of Ewing S100, 15% of triethyl citrate and 5% of talcum powder, and adding the three components into ethanol, and uniformly stirring. The coating machine is adopted to control the temperature to be about 45 ℃ and the coating machine is uniformly sprayed on the gelatin capsule shell to form a pH sensitive layer film which is dissolved only at the pH of 7.

2g of beta-cyclodextrin polymer or calcium pectin and hydroxypropyl methylcellulose are dissolved in 10 ml of dimethyl sulfoxide DMSO. Sucking 10ul of the solution, adding into the above capsule, and drying at 55 deg.C for 10 h. After cooling to room temperature, the colonic flora enzyme system activating layer is obtained.

The glutamic acid-717 resin in the embodiment 2 is filled into the prepared capsule to prepare the sustained-release glutamic acid preparation with colon-specific release.

The glutamic acid preparation is placed in gastric juice, intestinal juice and colon juice of a mouse, as shown in figure 2, and the capsule can resist the acid environment of the stomach and the middle upper end of the small intestine, and can be triggered and dissolved only by enzyme of flora peculiar to the colon part, so that the selectivity of the capsule to the colon part can be effectively enhanced.

Example 4 preparation of a colonic site-directed Release sustained Release citric acid formulation

In this example, 717 strongly basic anion exchange resin was used as the slow release carrier.

Pretreatment of i.717 resin

A 717 resin copolymer of styrene-divinylbenzene was used to encapsulate glutamic acid. It has a quaternary ammonium cationic group and active chloride ions. By substituting chloride ions attached to quaternary ammonium groups, drug ions can be loaded into the resin. Specifically, a quantity of 717 resin was washed by immersion in 50 ℃ deionized water to remove water soluble impurities, and then transferred to 95% ethanol with stirring to remove organic impurities. After washing with deionized water until no residual ethanol was present, it was dried under vacuum at 50 ℃. The predried 717 resin was immersed in a 0.1mol/l hydrochloric acid solution for 24h with constant stirring, then washed to neutrality with deionized water and dried to give an anion (Cl-) exchange resin.

ii, preparing a sustained release formulation of citric acid-717 resin

Purified citric acid (2.0g) was dissolved in 50mL of deionized water at room temperature, 3g of anion exchange resin was added, and the mixture was stirred at room temperature for 12 hours. And pouring out the supernatant, washing with water for a plurality of times, and naturally drying to obtain the resin loaded with the citric acid. 1.0g of resin loaded with 1.2g to 1.5g of citric acid was measured.

By adding citric acid-resin to the colon targeted release capsule prepared according to the method of example 3 or other commercially available formulations, a colon targeted release sustained release citric acid formulation is obtained.

As shown in FIG. 3, a graph showing the release results of the citric acid-resin added to the capsule for colon site-specific release prepared according to the method of example 3 shows that such a capsule can endure the acidic environment of the upper part of the stomach and small intestine and can be dissolved only by the enzyme of the flora specific to the colon site, thereby effectively enhancing the selectivity to the colon site.

The following experiments describe that the sustained-release citric acid preparation prepared by the present example is a preparation obtained by adding citric acid-resin to a colon-specific release capsule prepared according to the method of example 3.

EXAMPLE 5 colonic-targeted delivery of 7 alpha-dehydroxy inhibitor in animals

8 rats (SPF grade, body weight 220g-260g) were free to eat and drink water at room temperature 25 ℃. Stool samples were taken the day before drug intake. The colon-specific sustained release citric acid preparation (prepared in example 4) (prepared by using gelatin capsules of a size specific for small animals) is orally administered at a weight of 1g/Kg for intragastric administration for 4 weeks, and fecal samples are taken and tested for the contents of primary bile acid and secondary bile acid before and after treatment, with the results as shown in table 1:

TABLE 1

Therefore, the absolute quantity and the ratio of the secondary bile acid in the feces are obviously reduced after treatment, and the total bile acid is also obviously reduced.

After the experiment of the rats in the experimental group, 5 rats in a blank control group (SPF grade, weight 220g-260g, room temperature 25 ℃, free food and water) were anesthetized, and then the corresponding digestive tract tissue was taken to immediately detect pH, and the results are shown in Table 2:

TABLE 2

packet/pH	Stomach (stomach)	Jejunum	Ileum	Colon
					Blank control group	2.42±1.38	5.79±0.34	6.95±0.22	7.02±0.33
Colon sustained-release citric acid group	2.19±1.10	5.21±0.86	6.08±0.55	4.89±0.61

As can be seen, colon pH decreased significantly after treatment (P < 0.01).

Example 6 colonic targeted delivery of a sustained release 7 α -dehydroxy inhibitor

The influence of the slow-release aspartic acid (Asp), malic acid, acetic acid, butyric acid and other substances released in the colon in a positioning way on the pH value of the colon is respectively tested, the contents of the primary bile acid and the secondary bile acid in the fecal sample before and after treatment are respectively detected, the preparation method is the same as that of example 4, the test method is the same as that of example 5, and the results are as follows:

aspartic acid (Asp):

TABLE 3

ng/mg excrement	Absolute value before treatment	Absolute value after treatment	P value
				Primary bile acid
Cholic acid	689±171	301±186	<0.001
				Chenodeoxycholic acid	630±119	184±177	<0.001
Secondary bile acid
				Deoxycholic acid	1599±142	198±81	<0.001
Ursodeoxycholic acid	1206±294	174±130	<0.001
				Lithocholic acid	1572±502	138±106	<0.001

Malic acid:

TABLE 4

ng/mg excrement	Absolute value before treatment	After treatmentTo the value	P value
				Primary bile acid
Cholic acid	792±455	289±208	<0.001
				Chenodeoxycholic acid	511±205	360±165	<0.001
Secondary bile acid
				Deoxycholic acid	1175±386	101±55	<0.001
Ursodeoxycholic acid	1062±521	199±136	<0.001
				Lithocholic acid	1381±310	125±111	<0.001

Acetic acid:

TABLE 5

ng/mg excrement	Absolute value before treatment	Absolute value after treatment	P value
				Primary bile acid
Cholic acid	674±224	362±324	<0.001
				Chenodeoxycholic acid	435±410	383±251	<0.05
Secondary bile acid
				Deoxycholic acid	1944±805	282±140	<0.001
Ursodeoxycholic acid	1581±403	104±89	<0.001
				Lithocholic acid	1547±555	188±151	<0.001

Butyric acid:

TABLE 6

ng/mg excrement	Absolute value before treatment	Absolute value after treatment	P value
				Primary bile acid
Cholic acid	401±377	265±65	<0.001
				Chenodeoxycholic acid	979±645	438±354	<0.001
Secondary bile acid
				Deoxycholic acid	1730±659	452±215	<0.001
Ursodeoxycholic acid	1484±230	424±280	<0.001
				Lithocholic acid	1262±374	299±265	<0.001

It can be seen that similar experimental results can be achieved with the acids described above.

EXAMPLE 7 colonic targeted delivery of a sustained Release Glutamine formulation for the treatment of non-alcoholic steatohepatitis and liver cancer

SPF grade C57BL/6J mice were fed high fat diet (D12492, 60 kcal% fat) for 12 weeks one month after birth, with no restriction in diet and water. Mice were injected intraperitoneally with 200ug STZ (STZ in 0.1M citrate-HCl buffer, pH 4.5) on day 2 of birth. And 4 mice are taken at 12 weeks for anesthesia, then liver tissues are taken for pathological examination, and the mice are killed by dislocation of cervical vertebrae after the experiment is completed. The mice with non-alcoholic fatty liver disease and liver cancer which are successfully modeled are continuously divided into two groups for testing, one group recovers normal feed, and the other group is fed with 3% colon-specific release sustained-release glutamic acid preparation (prepared in example 3), and the diet and drinking water are not limited. Treatment was continued for a period of 12 weeks. 6 mice are respectively taken at 6 th and 12 th week of treatment, after anesthesia, liver tissue is taken for pathological examination, and after the experiment is completed, the mice are killed by dislocation of cervical vertebra. In the whole experiment process, indexes such as mouse tail blood, body weight and the like are collected once a week.

As a result, the mice treated with the colon-specific release sustained-release glutamic acid preparation had complete remission of nonalcoholic steatohepatitis and liver cancer. As shown in fig. 4: a B is a control group for 12 weeks after the establishment of 12 weeks non-alcoholic steatohepatitis-hepatic fibrosis-liver cancer model liver histopathology section C; D. and E is a sustained release glutamic acid preparation treatment group continuing colon-specific release for 12 weeks. The normal range for the ALT index to recover is also shown in fig. 5.

Example 8 study of diabetes model mice

The mouse model established in example 6 above is also suitable for the study of diabetes models. SPF grade C57BL/6J mice were fed high fat diet (D12492, 60 kcal% fat) for 12 weeks one month after birth, with no restriction in diet and water. Mice developed type II diabetes by intraperitoneal injection of 200ug STZ (STZ in 0.1M citrate buffered saline, pH4.5) once on day 2 of birth. One group of 6 normal feeds is fed with 3% colon-specific release sustained-release glutamic acid preparation (prepared in example 3), and the diet and drinking water are not limited. Treatment was continued for a period of 12 weeks. In the whole experiment process, indexes such as mouse tail blood, body weight and the like are collected once a week.

The results are shown in fig. 6, which shows that the colon-specific release sustained-release glutamic acid preparation can significantly improve blood glucose metabolism, and the fasting blood glucose and the insulin resistance index are recovered to normal levels after treatment, which are significantly different from those of the control group.

Example 9 prevention and treatment of colorectal adenoma and colorectal carcinoma with a colonic targeted release sustained release citric acid formulation

SPF grade C57BL/6J mice 6 weeks old are free to eat and drink at room temperature about 25 ℃. Diazomethane (AOM, 10mg/kg) was injected intraperitoneally twice a week for 6 consecutive weeks. Divided into two groups, drug intervention was initiated simultaneously at the beginning of intraperitoneal AOM injection. Blank control group; the treatment group was fed with a colon specific release sustained release citric acid formulation (prepared in example 4) 500 mg/Kg/day. Mice were sacrificed by anesthesia and cervical dislocation at week 15 and week 30, and colon tissues were analyzed. The number of adenomas is shown in table 7:

TABLE 7

Group of	Number of adenomas/single	Size of adenoma (mm)
			Control group (15)Mouse)	7.6±2.5	3.8±1.5
Citric acid treatment group (15 mice)	1.2±0.8	1.1±0.5

It can be seen that the colon-specific release sustained-release citric acid preparation treatment group significantly reduced the formation of adenomas compared with the control group, with P < 0.001.

FIG. 7 shows that 30% of mice in the placebo group exhibited High Grade Intraepithelial Neoplasia (HGIN) and even carcinoma in situ, whereas the treated group exhibited a low number of adenomas and no high grade intraepithelial neoplasia or carcinoma in situ.

The person skilled in the art will be able to identify and know that: the above examples are merely one preferred embodiment known at present, and are not limited to the specific formulation forms listed in the examples, as long as they can deliver the molecules inhibiting the 7 α -dehydroxylation of intestinal flora and the formation of secondary bile acids through the colon-specific delivery system.

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