CN107684550B

CN107684550B - Diabetes treatment product and preparation and application thereof

Info

Publication number: CN107684550B
Application number: CN201610629475.6A
Authority: CN
Inventors: 徐天宏
Original assignee: Individual
Current assignee: Individual
Priority date: 2016-08-03
Filing date: 2016-08-03
Publication date: 2020-04-10
Anticipated expiration: 2036-08-03
Also published as: CN107684550A

Abstract

The invention belongs to the field of drug research and development, and particularly relates to a diabetes treatment product and preparation and application thereof. The diabetes treatment product comprises an oral fixed-release preparation which is released at the tail end of ileum and colon in a fixed way, and the oral fixed-release preparation contains nutrient components. After extensive and intensive research, the invention firstly discovers that by adopting the diabetes treatment product, nutrient components are positioned at the tail end of ileum and colon for release, or the intestinal mucosa adhesive polymer is positioned in duodenum for release in a combined manner, so that the intestinal mucosa adhesive polymer is adhered to the duodenum mucosa to form shielding, the absorption of duodenum is prevented, and the effect of metabolic surgery can be simulated and achieved more perfectly.

Description

Diabetes treatment product and preparation and application thereof

Technical Field

The invention belongs to the field of drug research and development, and particularly relates to a diabetes treatment product and preparation and application thereof.

Background

Type 2 diabetes currently affects the health of more than 4.15 billion people worldwide, and more seriously, the diabetes-related mortality rate will rise by more than 50% in the next 10 years. According to data released by the committee of health care in China, the prevalence rate of diabetes mellitus type 2 of adults in China at present reaches 11.4 percent, and rises to the first world and reaches 1.4 hundred million people, and complications caused by diabetes mellitus, such as heart disease, renal failure, blindness, foot ulcer and the like, also rise linearly. In addition, 20% of the population in China is in the pre-diabetic state, and the sugar metabolism of the population is already abnormal.

Type 2 diabetes mainly ranges from insulin resistance with inadequate insulin secretion to insulin resistance with inadequate insulin secretion. Type 1 diabetes is mainly caused by B cell destruction and absolute insulin deficiency, and most of them are autoimmune diseases.

Meanwhile, nearly one third of people worldwide are overweight or obese. In China, obesity has increased explosively. At present, about 43.6% of Chinese people are overweight or obese, i.e., 6 billion. Obesity has become a major problem worldwide. Obesity can lead to a range of complications or related diseases that in turn affect our longevity or lead to a reduction in quality of life. Diabetes is often associated with obesity. Obesity accounts for 41.5% of the population with diabetes mellitus, a causative factor of diabetes mellitus.

The current treatment of type 2 diabetes is primarily drug therapy, with a wide variety of drugs, such as biguanides (e.g., metformin), sulfonylureas (e.g., glimepiride, glyburide, gliclazide, and gliquidone), thiazolidinediones (e.g., rosiglitazone and pioglitazone), benzoic acid derivatives, insulin secretagogues, α -glucosidase inhibitors (e.g., acarbose and voglibose), GLP-1 analogs, DPPIV, and the like.

However, these drugs have certain side effects, for example, many patients cannot use them due to gastrointestinal side effects when taking metformin, and DPPIV drugs have been found to increase the risk of heart failure; after the diabetes mellitus medicines are taken for a long time, drug resistance is easy to generate, so that one medicine cannot control blood sugar, and needs to be changed or combined with other medicines, and finally, the diabetes mellitus is transited to insulin-dependent diabetes mellitus; the medicines are all required to be discharged by metabolism of liver and kidney, and cannot be used by patients with liver and kidney insufficiency; it is obvious that these drugs and insulin can only control blood sugar, and are temporary-curing and permanent-curing, and need to be used for life.

The development of various novel diabetes treatment drugs has been a development focus of various pharmaceutical companies and research institutions.

The first general guidelines for the treatment of type 2 diabetes mellitus by metabolic surgery jointly established by multiple organizations in 24.5.2016- -the International organization for diabetes mellitus has published 45 International organizations such as the American Diabetes Association (ADA), the International Diabetes Federation (IDF), the British diabetes Association (DUK), the Chinese medical society diabetes mellitus (CDS) and the India diabetes mellitus (DI) for the treatment of type 2 diabetes mellitus by metabolic surgery in combination with the guidelines. Metabolic surgery should become a treatment strategy for type 2 diabetes. This guideline is the most fundamental change that has occurred in the field of treatment of type 2 diabetes in the last century.

According to statistics of clinical researches on metabolic operations in various parts of the world in the past decades, 80% -95% of diabetes patients do not need any drug treatment or special diet after the metabolic operations, and blood sugar, insulin and glycosylated blood protein are recovered to be normal. The follow-up of the patients for 5 to 10 years after the operation discovers that more than 80 percent of the patients with type II diabetes recover the normal blood sugar, no drug treatment or diet restriction is needed, the symptoms of the diabetes can be obviously relieved, even completely cured, and can be maintained for a long time, and the effect of the gastric bypass operation which changes the gastrointestinal tract sequence is obviously better than that of the simple gastric volume restriction operation, namely, the metabolic operation becomes the only treatment strategy which can cure the diabetes.

The core of the metabolic operation treatment method represented by the gastric bypass operation is to change the normal physiological flow direction of food, so that after the operation for curing diabetes, the food in the upper digestive tract is bypassed and does not pass through the two ends of the stomach, the duodenum and the upper end of the jejunum any more.

Diet loss, weight loss are not the major cause of metabolic surgery for diabetes, and glycemic control and improvement of diabetic symptoms are far earlier than weight loss. The current main research shows that the metabolism operation changes the endocrine hormone of the gastrointestinal tract and is the main mechanism for long-term cure of the postoperative diabetes. It is clear, however, that metabolic surgery is a duplicative surgery, requiring a high skill set by the surgeon, and also presenting a number of risks of complications, such as internal bleeding, abdominal pain, colics, constipation or diarrhea, dumping syndrome, infections, vitamin deficiencies, etc.

K cells are mainly distributed in the upper small intestine and secrete insulin resistance factors such as GIP (Glucose-dependent insulin-releasing peptide) and the like after being stimulated by food nutrients such as sugar, protein and fat. The L cells are mainly distributed on the lower segment of small intestine and large intestine mucosa, and if stimulated by glucose, amino acid, fatty acid and the like, the L cells can generate GLP-1 (Glucagon-like peptide-1), GLP-2, PYY and other endocrine hormones, thereby enhancing insulin secretion, reducing insulin resistance and improving glycometabolism.

GIP is an incretin synthesized and secreted by K cells in the upper segment of the duodenum and jejunum, and is mainly secreted by food stimulation. Excessive secretion thereof results in the deposition of lipids in peripheral tissues (liver, muscle, etc.) and islet B cells, thereby causing impaired insulin resistance and secretory function, and inhibition of GIP secretion can significantly improve obesity-related type 2 diabetes.

GLP-1 is an incretin synthesized and secreted by L cells of the distal ileum and the large intestine, glucose, amino acid and fat in food are the most main nutrient substances for stimulating the release of the incretin, and GLP-1 can promote glycogen synthesis and lipolysis, inhibit gastric emptying and glucagon secretion, increase the gene expression of insulin and the synthesis of insulin precursors, and promote B cell proliferation and inhibit apoptosis. Many drug developments are focused on the treatment of diabetes, obesity and other diseases by using GLP-1, but the natural GLP-1 of a human body is very unstable and can be degraded by dipeptidyl peptidase IV (DPP IV), and the half-life period is only 1-2 minutes. Inhibition of GLP-1 degradation by GLP-1 receptor agonists, either by direct synthesis of long-acting GLP-1 analogs, or by small molecules, is a hot spot in drug development.

GLP-2 (glucagon-like peptide-2 glucose-like peptide-2) is a 33 amino acid polypeptide secreted by L cells. GLP-2 is a newly discovered specific growth factor for intestinal epithelium, and has various effects on gastrointestinal tract, including promoting growth and development of normal small intestine, protecting and repairing damaged intestinal mucosa in various intestinal diseases, inhibiting secretion of gastric acid and gastrointestinal motility, and increasing blood supply of intestinal tract.

PYY is also a 36 amino acid polypeptide secreted by L cells. It can reduce appetite.

Oxyntomodulin (OXM) is a 37 amino acid polypeptide secreted by L cells and has the effects of reducing appetite and inhibiting gastric acid secretion.

After metabolic surgery, the upper digestive tract including the duodenum does not receive food stimulation any more, and K cells mainly distributed on the mucous membrane of the upper digestive tract do not secrete insulin resistance factors such as GIP (insulin resistance factor) and the like, so that the insulin resistance phenomenon of the body is eliminated. Meanwhile, food which is not completely digested can enter the middle and lower digestive tracts earlier, so that L cells distributed in the mucous membranes of the lower digestive tracts can be stimulated to secrete hormones such as PYY, GLP1, GLP2 and OXM. These hormones have a common effect: firstly, directly reducing blood sugar; second, apoptosis of islet cells is eliminated; third, islet cell proliferation can be stimulated. These hormones may also protect pancreatic islet cells from damage such as glucotoxicity, lipotoxicity and other inflammatory conditions; fourth, reduce appetite and reduce food intake.

Disclosure of Invention

In order to overcome the problems in the prior art, the invention aims to provide a diabetes treatment product and preparation and application thereof.

In order to achieve the above objects and other related objects, the present invention adopts the following technical solutions:

in a first aspect the present invention provides a diabetes treatment product comprising an oral targeted release formulation for targeted release in the terminal ileum and colon, the oral targeted release formulation comprising a nutritional ingredient.

The terminal ileum refers to the part of the ileum close to the colon.

Preferably, the nutritional ingredients are selected from: any one or combination of more of carbohydrate, fat, or protein degradation products.

Preferably, the nutritional ingredients are selected from: any one or more of acetic acid, propionic acid, butyric acid, bile acid, long chain fatty acid, glucose, glutamine or arginine.

Preferably, the nutrient components are: a combination of propionic acid and arginine, a combination of propionic acid and glutamine, a combination of acetic acid and arginine, a combination of acetic acid and glutamine, a combination of butyric acid and arginine, a combination of butyric acid and glutamine, a combination of bile acid and arginine, a combination of bile acid and glutamine, a combination of long chain fatty acid and arginine, or a combination of long chain fatty acid and glutamine.

The long chain fatty acid contains 14 or more carbon atoms.

Preferably, in each combination, the mass ratio of the two nutrient components is (1-10): (1-10).

In the embodiment of the invention, in each combination, the mass ratio of the two nutrient components is 1: 1.

The invention finds that the two nutrient components in each combination are used together, and have a synergistic promoting effect when stimulating L cells to release the gastrointestinal hormone, so that the effect of 0.5+0.5 more than 1 can be achieved.

Preferably, the nutritional ingredients are encapsulated in an intestinal mucoadhesive polymer.

The intestinal mucoadhesive polymers are state of the art.

For example, Curr Drug Deliv 2015; 12(2):139-56.

A Mucoadsive polymeric platform for a drug delivery; a comprehensive review, displaced Monomers and Polymers 12(2009),483-495Polymers in multimedia Drug Delivery System: A Brief Note.

The intestinal mucosa adhesive polymer is preferably a polymer with relatively strong intestinal mucosa adhesiveness, no toxic or side effect and good biocompatibility.

Preferably, the intestinal mucosa-adhesive polymer is selected from any one or more of hydrophilic polymers, hydrogel-like polymers, thiolated polymers, and lectin polymers.

Preferably, the mucoadhesive enteric polymer is selected from any one or more of chitosan, polystyrene, carbomer, carbopol-cysteine, polyacrylic acid-homocysteine, alginate-cysteine, polymethacrylic acid-cysteine, sodium carboxymethylcellulose-cysteine in combination.

Further preferably, the intestinal mucosa-adhesive polymer is selected from thiolated chitosan or a combination of any one or more of chitosan-catechol.

Preferably, the mucoadhesive enteric polymer may be prepared in the form of liquid microspheres. After the release at the tail end of ileum and colon, the tightness and the persistence of nutrient components and intestinal mucosa can be enhanced, and the absorption of the nutrient components is facilitated.

Preferably, the oral targeted release formulation for targeted release in the terminal ileum and colon is an enteric coated release formulation.

Further preferably, the oral localized release formulation for localized release in the terminal ileum and colon is a pH-sensitive enteric coated release formulation.

The polymer can be selected to be soluble at a suitable pH range, as desired. Ensures that the nutrient components are positioned at the tail end of ileum and released in colon.

In a preferred embodiment of the invention, the pH sensitive enteric coating is a polyacrylic resin Eudragit S100.

Preferably, the diabetes treatment product also comprises an oral duodenal positioning release preparation, and the oral duodenal positioning release preparation contains an intestinal mucosa adhesive polymer.

Preferably, the oral duodenal-site release formulation locates and releases the intestinal mucoadhesive polymer to the duodenum, and the intestinal mucoadhesive polymer adheres to the duodenal mucosa to form a barrier to prevent absorption in the duodenum.

The intestinal mucoadhesive polymers are state of the art. For example, Curr Drug Deliv 2015; 12(2):139-56.

Preferably, the intestinal mucosa-adhesive polymer is selected from any one or a combination of hydrophilic polymers, hydrogel-like polymers, thiolated polymers, or lectin polymers.

Preferably, the mucoadhesive enteric polymer is selected from any one or more of chitosan, polystyrene, carbomer, carbopol-cysteine, polyacrylic acid-homocysteine, alginate-cysteine, polymethacrylic acid-cysteine or sodium carboxymethylcellulose-cysteine in combination.

Preferably, the mucoadhesive enteric polymer may be prepared in the form of liquid microspheres. After the duodenum is released, the contact area with the intestinal mucosa can be increased, and the use is convenient.

Preferably, the oral duodenal localized release formulation is an enteric coated release formulation.

Further preferably, the oral duodenal localized-release preparation is a release preparation coated with a pH-sensitive enteric coating.

Polymers that dissolve in a suitable pH range may be selected as desired, for example, polymers that achieve rapid release at pH greater than 5.0 and even greater than 3.0 may be selected. Ensuring that the intestinal mucosa adhesive polymer is positioned in the duodenum for release.

In a preferred embodiment of the invention, hydroxypropyl methylcellulose acetate maleate (HPMCAM) is used as the enteric coating.

Preferably, the diabetes treatment product is for the treatment of type I diabetes, type II diabetes, pre-diabetes, obesity, appetite control, metabolic syndrome or polycystic ovary syndrome.

Preferably, the diabetes treatment product is a metabolic surgery treatment replacement product.

Patients using the diabetes treatment product can no longer be treated with metabolic surgery.

Further preferably, the diabetes treatment is a gastric bypass surgery treatment substitute.

Patients using the diabetes treatment product can no longer be treated by gastric bypass surgery.

In a second aspect of the invention there is provided the use of a diabetes treatment product as hereinbefore described in the manufacture of a medicament for the treatment of type I diabetes, type II diabetes, pre-diabetes, obesity, appetite control, metabolic syndrome or polycystic ovary syndrome.

Preferably, the therapeutic agent is a metabolic surgery replacement agent.

Patients using the therapeutic agent may no longer be treated by metabolic surgery.

Further preferably, the therapeutic drug is a gastric bypass surgery substitute drug.

Patients using the therapeutic agent may no longer be treated by gastric bypass surgery.

In a third aspect of the invention, there is provided a method of treating type I diabetes, type II diabetes, pre-diabetes, obesity, appetite control, metabolic syndrome or polycystic ovary syndrome, comprising the steps of: administering a therapeutically effective amount of the aforementioned diabetes treatment product to a corresponding subject.

Preferably, the mode of administration is oral.

Preferably, the method is an alternative to metabolic surgery.

Further preferably, the method may replace gastric bypass surgery.

By "therapeutically effective amount" is meant any amount that results in improved treatment, cure, prevention, or amelioration of a disease, disorder, or side effect, or a reduction in the rate of progression of a disease or disorder, as compared to a corresponding subject that has not received such amount. The term also includes within its scope an amount effective to enhance normal physiological function. A therapeutically effective amount will produce a "therapeutic effect".

A therapeutically effective amount of the delayed release formulation 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 case, the effective amount of the delayed release formulation of the invention for treating a patient suffering from a diabetic or overweight condition and related conditions should, in general, be 0.01 to 10g/kg of the body weight of the recipient (mammal)/day. More typically, an effective amount should be 30 to 300mg/kg body weight/day. Thus, for an adult mammal of 70kg body weight, the actual amount per day is typically from 2.1 to 21 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.

Compared with the prior art, the invention has the following beneficial effects:

after extensive and intensive research, the invention firstly discovers that by adopting the diabetes treatment product, nutrient components are positioned at the tail end of ileum and colon for release, or the intestinal mucosa adhesive polymer is positioned in duodenum for release in a combined manner, so that the intestinal mucosa adhesive polymer is adhered to the duodenum mucosa to form shielding, the absorption of duodenum is prevented, and the effect of metabolic surgery can be simulated and achieved more perfectly.

Drawings

FIG. 1: in example 1, changes in GLP-1 levels in blood were analyzed.

FIG. 2: in example 1, PYY level changes in blood were analyzed.

FIG. 3: in example 1, changes in insulin levels in blood were analyzed.

FIG. 4: in example 1, the glucose level in blood was analyzed.

FIG. 5: in example 2, component a is a microsphere prepared under an electron microscope.

FIG. 6: in example 2, component B is a microsphere prepared under an electron microscope.

FIG. 7: in example 2, changes in GLP-1 levels in blood were analyzed.

FIG. 8: in example 2, changes in insulin levels in blood were analyzed.

FIG. 9: the CC microspheres and TC microspheres were tested for mucosal adhesion, respectively.

Detailed Description

The present inventors have found that various diseases including diabetes, obesity, etc. can be treated by releasing some naturally occurring nutrients not when they pass through the upper digestive tract (stomach, duodenum, jejunum) but when they reach the terminal ileum and colon. The action and the treatment mechanism are similar to those of gastric bypass surgery, namely, nutrient substances are prevented from stimulating K cells of upper digestive tract (stomach, duodenum and jejunum), and secretion of insulin resistance factors such as GIP (Glucose-dependent insulin release peptide) is inhibited; meanwhile, a large amount of L cells of the digestive tract are stimulated to generate endocrine hormones such as GLP-1 (Glucagon-like peptide-1), GLP-2, PYY and the like, so that the secretion of insulin is enhanced, the resistance of insulin is reduced, and the sugar metabolism is improved.

Natural nutrient components include fatty acid, monosaccharide or polysaccharide, and amino acid; the preferable specific nutrient components include one or two or more selected from acetic acid, propionic acid, butyric acid, bile acid, long-chain fatty acid, glucose, glutamine or arginine. The individual is administered sufficient stimulating nutrients or a combination thereof, using a formulation capable of delivering it to the terminal ileum and colon, to cause the release of oxyntomodulin from L cells of the lower digestive tract of the individual while achieving the desired result and avoiding stimulation of K cells of the upper digestive tract.

There are a number of formulations available in the prior art that can deliver a substance to the terminal ileum and colon for release. The oral positioning release preparation which is positioned and released at the tail end of the ileum and the colon can keep complete in the stomach after being orally taken, and can release substances according to the design requirement after entering the tail end of the ileum and the colon, 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. Because of the effect of gastric emptying time, the use of controlled release formulations alone for lag time does not necessarily achieve complete terminal ileal and colonic targeted release, and controlled release techniques can be combined with the use of enteric coating techniques to ensure that the substance is released only in the terminal ileum and colon.

Examples of oral targeted release formulations that are targeted for release at the terminal ileum and colon include, but are not limited to, those described in U.S. patent nos. 749307; 3,845,770; 3,916,899; 3,536,809, respectively; 3,598,123, respectively; 4,008,719, respectively; 5,674,533, respectively; 5,059,595, respectively; 5,591,767, respectively; 5,120,548, respectively; 5,073,543, respectively; 5,639,476, respectively; 5,354,556, respectively; and those described in U.S. Pat. No. 5,733,566. Such dosage forms may be used to provide the desired release profile using hydroxypropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres or combinations thereof in varying proportions to provide release of one or more active ingredients in the terminal ileum and colon.

Preferably, we use the current use of relatively common pH sensitive coatings to prepare oral targeted release formulations for localized release in the terminal ileum and colon to deliver naturally occurring nutritional compounds to the terminal ileum and colon. The oral localized release formulation for localized release in the terminal ileum and colon comprises an oral dosage form having a core containing a therapeutically effective amount of a nutritional compound and an enteric coating surrounding the core. The pH sensitivity of the enteric coating may be set to a delay period of at least about 5 to 10 minutes after sustained contact with a pH of 5.0,5.5,6.0,6.5 or 7.0, respectively, more preferably to a delay period of at least about 15 or 20 minutes after contact with the desired pH, and still more preferably to a delay period of at least about 25 or 30 minutes after contact with the desired pH in the terminal ileum and colon. In a particularly preferred embodiment, the formulation ensures the re-release of nutrients after reaching the terminal ileum and colon through the upper digestive tract, wherein the slow release (extendedrelease) is initiated after a lag phase at a pH of at least about 6.0, more preferably at least about 6.5, and still more preferably at 6.8 or 7.0, prolonging the time of release of these nutrients at the terminal ileum and colon.

Conditions treated by ingestion of such formulations include, but are not limited to, type I diabetes, type II diabetes, pre-diabetic obesity, appetite control, metabolic syndrome, and polycystic ovary syndrome.

In the treatment of type II diabetes, the formulations of the invention may be combined with one or more pharmaceutically active agents, such as metformin, sulfonylureas, such as glibenclamide and glipizide, repaglinide, nateglinide, thiazolidinediones, such as rosiglitazone and pioglitazone, acarbose, miglitol, exenatide (exenatide), pramlintide and insulin.

In addition, while the nutrient components are positioned at the tail end of the ileum and the colon for release, the intestinal mucosa adhesive polymer is positioned in the duodenum for release, adheres to the duodenal mucosa to form a shield, prevents the absorption of the duodenum, and can perfectly simulate and achieve the effect of metabolic surgery. The release is carried out by targeting the intestinal mucoadhesive polymer to the duodenum, forming a temporary barrier at the duodenal mucosa, lasting from hours to tens of hours, blocking food from contact with the duodenal mucosa. Avoid the stimulation of K cells of duodenum by nutrient substances and inhibit the secretion of insulin resistance factors such as GIP (glucose-dependent insulin-releasing peptide).

There are a number of formulations currently available in the art that can deliver a substance to the duodenum for release. The oral duodenum positioning release preparation is kept complete in the stomach after being orally taken, and can release substances according to design requirements after entering the duodenum, 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. Due to the effect of gastric emptying time, the application of controlled release formulations alone may not completely achieve a duodenal site-specific release with a lag time, and the controlled release time technique may be combined with the enteric coating technique to ensure that the substance is released only in the duodenum.

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 only, and is not intended to limit the scope of the present invention. Test methods in which specific conditions are not specified in the following examples are generally carried out under conventional conditions or under conditions recommended by the respective manufacturers.

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.

Unless otherwise indicated, the experimental methods, detection methods, and preparation methods disclosed herein all employ techniques conventional in the art of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related arts. These techniques are well described in the literature, and may be found in particular in the study of the MOLECULAR CLONING, Sambrook et al: a LABORATORY MANUAL, Second edition, Cold Spring harbor LABORATORY Press, 1989and Third edition, 2001; ausubel et al, Current PROTOCOLS Inmolecular BIOLOGY, John Wiley & Sons, New York, 1987and periodic updates; the series METHODS IN ENZYMOLOGY, Academic Press, San Diego; wolffe, CHROMATINSTRUCUTURE AND FUNCTION, Third edition, Academic Press, San Diego, 1998; (iii) Methods Inenzymolygy, Vol.304, Chromatin (P.M. Wassarman and A.P.Wolffe, eds.), academic Press, San Diego, 1999; and METHODS IN MOLECULAR BIOLOGY, Vol.119, chromatography protocols (P.B.Becker, ed.) Humana Press, Totowa, 1999, etc.

Example 1

This example prepares a diabetes treatment product containing a B-component, which is an oral targeted release formulation containing nutritional ingredients that is targeted for release in the terminal ileum and colon.

This example refers to the prior art for preparing oral targeted release formulations for targeted release in the terminal ileum and colon for targeted release of nutrients in the terminal ileum and colon. Oral localized release formulations prepared for localized release in the terminal ileum and colon are known in the art, for example, reference may be made to the technology described in the invention patent application No. 201480003932. X.

Specifically, Thiolated Chitosan microspheres were prepared using Thiolated Chitosan (TC) as a support material in this example. Then dissolving or dispersing the nutrient components in the thiolated chitosan microsphere entity to prepare the thiolated chitosan microsphere coated with the nutrient components. Then the thiolated chitosan microspheres coated with the nutrient components are coated in polyacrylic resin Eudragit S100 which is enteric carrier material, and the oral fixed-release preparation which is fixed-release at the tail end of ileum and colon is prepared. It is well known to those skilled in the art that thiolation can be applied to chitosan in many locations, all of which have strong mucoadhesive properties. Therefore, the present embodiment only uses one specific form of thiolated chitosan as an example, but is not limited to the thiolated chitosan listed. The specific thiolated chitosan used in this example was: chitosan-4-mercaptobutylamine.

More specifically, (1) preparing a microsphere crosslinking solution of TC coated with propionic acid and glutamine as nutrient components, comprising the steps of:

240mg of TC was dissolved in 12ml of a 1% acetic acid solution to obtain an acidic polymer solution, and 30mg of arginine and 30mg of propionic acid were dispersed in the acidic polymer solution. Then 50ml of liquid paraffin containing 1 percent of Span 80 is added and fully stirred; adding a toluene solution saturated with formaldehyde, fully stirring, adding a toluene solution saturated with glutaraldehyde, and fully stirring for 20 hours. And (3) cleaning the generated microspheres with normal hexane for several times, dehydrating with acetone, and sucking at room temperature to obtain thiolated chitosan microspheres wrapped with nutrient components.

(2) Coating the microspheres obtained in the step (1) by using Eudragit S100: 500mg Eudragit S100 was dissolved in ethanol: in a mixed solvent of acetone (ethanol: acetone volume ratio of 2: 1). 50mg of TC microspheres containing nutrients were dispersed in 10ml of the previous solution. 70ml of liquid paraffin containing 1% Span 80 was added, sufficiently stirred for 3 hours, and the organic solvent was volatilized. Washing with n-hexane for several times, and sucking at room temperature to obtain oral fixed-release preparation for fixed-release at terminal of ileum and colon.

It will be understood and appreciated by those skilled in the art that the oral targeted release formulation for targeted release in the terminal ileum and colon is not limited to the specific formulation forms listed in the examples, as long as it is an oral formulation capable of targeted release of nutrients in the terminal ileum and colon.

The oral fixed-release preparation which is released in the tail end of ileum and colon in a fixed way and is the same as the above is adopted to respectively prepare the effective component of (1) arginine; (2) (ii) glutamine; (3) propionic acid; (4) oral targeted release formulations of arginine and propionic acid.

The oral administration positioning release preparation is respectively (1) arginine oral administration positioning release tablets, and each arginine oral administration positioning release tablet contains 500mg arginine; (2) glutamine oral targeted release tablets, each glutamine oral targeted release tablet containing 500mg glutamine; (3) alanine oral targeted release tablets, each containing 500mg propionic acid; (4) each compound oral positioning release tablet contains 500mg arginine and 500mg propionic acid.

Each of the above oral targeted release formulations was targeted for release in the terminal ileum and colon.

15 overnight fasted insulin independent type 2 diabetic patients were divided into 5 groups in the early morning, and 3 patients were randomized into one group, and each group was administered the delayed sustained release formulation described above:

4 hours after administration, the patients received an oral glucose tolerance test. Blood was collected at the following time points: 30, 0, 15, 30, 60, 90, 120 minutes.

Blood is analyzed for the levels of hormones such as GLP-1, PYY, insulin, glucose, etc. Several of these four groups of patients showed significant improvement in the index compared to the control group with oral placebo (blank oral targeted release tablets without any nutrients, targeted release in the terminal ileum and colon). However, under the same experimental conditions, the effect of taking 2 compound oral targeted-release tablets in the fourth group on the treatment of the independent type 2 diabetes is better than that of taking 4 arginine oral targeted-release tablets and 4 alanine oral targeted-release tablets separately.

That is, compared with arginine alone as a nutritional ingredient and propionic acid alone as a nutritional ingredient, arginine and propionic acid are used together as the nutritional ingredient, the same oral targeted release preparation technology is adopted to position the nutritional ingredient to the terminal of the ileum and the colon for release, and arginine and propionic acid are used together as the nutritional ingredient, so that arginine and propionic acid have good synergistic effect on the L cells of the lower digestive tract to release gastrointestinal hormone, and the treatment effect of 0.5+0.5 to more than 1 can be achieved.

In addition, in this example, with reference to the above experimental method, the following were studied:

the acetic acid and the arginine are jointly used as nutrient components, compared with the single acetic acid and the single arginine, the same oral administration positioning release preparation technology is adopted to position the nutritional components to the tail end of the ileum and the colon for release, and the result shows that the acetic acid and the arginine jointly used as the nutrient components have good synergistic effect on the L cells of the lower digestive tract to release the gastrointestinal hormone, and the treatment effect of 0.5+0.5 to more than 1 can be achieved.

The acetic acid and the glutamine are jointly used as nutrient components, compared with the acetic acid alone as the nutrient component and the glutamine alone as the nutrient component, the same oral administration positioning release preparation technology is adopted to position the nutrient components to the tail end of the ileum and the colon for releasing, and the result shows that the acetic acid and the glutamine jointly used as the nutrient components have good synergistic effect on the L cells of the lower digestive tract to release the gastrointestinal hormone, and the treatment effect of 0.5+0.5 to more than 1 can be achieved.

Propionic acid and glutamine are jointly used as nutrient components, compared with propionic acid alone as the nutrient components and glutamine alone as the nutrient components, the propionic acid and the glutamine are positioned to the tail end of the ileum and the colon to release by adopting the same oral administration positioned release preparation technology, and the result shows that the propionic acid and the glutamine jointly used as the nutrient components have good synergistic effect on the L cells of the lower digestive tract to release the gastrointestinal hormone, and the treatment effect of 0.5+0.5 to more than 1 can be achieved.

The butyric acid and the glutamine are jointly used as nutrient components, and compared with the butyric acid alone as the nutrient component and the glutamine alone as the nutrient component, the butyric acid and the glutamine are positioned to the tail end of the ileum and the colon for releasing by adopting the same oral administration positioning release preparation technology, and the result shows that the butyric acid and the glutamine jointly used as the nutrient components have good synergistic effect on the L cells of the lower digestive tract to release the gastrointestinal hormone, and the treatment effect of 0.5+0.5 to more than 1 can be achieved.

The butyric acid and the arginine are jointly used as nutrient components, and compared with the single butyric acid and the single arginine, the butyric acid and the arginine are adopted as nutrient components, and the same oral administration positioning release preparation technology is adopted to position the butyric acid and the arginine to the tail end of the ileum and colon for release, and the result shows that the butyric acid and the arginine jointly used as the nutrient components have good synergistic effect on the L cells of the lower digestive tract to release the gastrointestinal hormone, and the treatment effect of 0.5+0.5 to more than 1 can be achieved.

The combination of bile acid and arginine is adopted as the nutrient component, the combination of bile acid and glutamine is adopted as the nutrient component, the combination of long-chain fatty acid and arginine is adopted as the nutrient component, or the combination of long-chain fatty acid and glutamine is adopted as the nutrient component, so that the curative effect of 0.5+0.5 more than 1 is achieved.

The present invention is not particularly limited with respect to the mass ratio between the two nutritional components in each combination. In each combination, the mass ratio of the two nutrient components is (1-10): (1-10) in any form.

In addition, this example also refers to the above preparation method and experimental method, and uses chitosan-catechol as carrier material to prepare chitosan-catechol microspheres. Then dissolving or dispersing the nutrient components in the chitosan-catechol microsphere entity to prepare the chitosan-catechol microsphere coated with the nutrient components. Then the chitosan-catechol microspheres coated with the nutrient components are coated in polyacrylic resin Eudragit S100 which is enteric carrier material, and the oral fixed-release preparation which is released at the tail end of ileum and colon fixed-release is prepared. Has good treatment effect.

Example 2

This example prepares a diabetes treatment product containing a component a and a component B, which are oral site-directed release formulations that are site-directed released in the terminal ileum and colon (same as example 1); the component A is an oral duodenal fixed-release preparation which contains an intestinal mucosa adhesive polymer.

This example refers to the prior art for preparing oral duodenal site-specific delivery formulations that deliver a site-specific intestinal mucoadhesive polymer to the duodenum, which adheres to the duodenal mucosa to form a barrier that prevents duodenal absorption.

Specifically, Thiolated Chitosan microspheres were prepared using Thiolated Chitosan (TC) as the intestinal mucosal adhesive polymer. Then the thiolated chitosan microspheres are wrapped in hydroxypropyl methyl cellulose acetate maleate (HPMCAM) to prepare the oral duodenal targeted release preparation. It is well known to those skilled in the art that thiolation can be added to chitosan at many locations, but with little effect. Therefore, the present embodiment only uses one specific form of thiolated chitosan as an example, but is not limited to the thiolated chitosan listed. The specific thiolated chitosan used in this example was: chitosan-4-mercaptobutylamine.

More specifically, (1) preparing a microsphere crosslinking solution for preparing TC includes the steps of: dissolving 300mg TC in 12ml of 1% acetic acid solution, adding 50ml of liquid paraffin containing 1% Span 80, and stirring thoroughly; adding a toluene solution saturated with formaldehyde, fully stirring, adding a toluene solution saturated with glutaraldehyde, and fully stirring for 20 hours. The resulting microspheres were washed several times with n-hexane, dehydrated with acetone, and blotted dry at room temperature.

(2) Coating the microspheres obtained in the step (1) by using hydroxypropyl methyl cellulose acetate maleate (HPMCAM):

200mg of HPMCAM was dissolved in 5ml of a mixed solvent of ethanol and acetone (ethanol: acetone in a volume ratio of 2:1), 40mg of TC crosslinked microspheres and 20mg of aluminum stearate were added, 20ml of liquid paraffin was added dropwise, and the mixture was sufficiently stirred. After completely evaporating off acetone and ethanol, washing the microspheres with n-hexane for several times, and sucking at room temperature for later use to obtain the oral duodenal targeted release preparation, wherein an electron microscope picture of the microspheres is shown in fig. 5.

The component B is the same as the component in example 1, and the specific preparation method comprises the following steps:

(1) preparing a microsphere cross-linked solution of TC coated with nutritional components propionic acid and glutamine, comprising the steps of:

(2) Coating the microspheres obtained in the step (1) by using Eudragit S100: 500mg Eudragit S100 was dissolved in ethanol: in a mixed solvent of acetone (ethanol: acetone volume ratio of 2: 1). 50mg of TC microspheres containing nutrients were dispersed in 10ml of the previous solution. 70ml of liquid paraffin containing 1% Span 80 was added, sufficiently stirred for 3 hours, and the organic solvent was volatilized. Washing with n-hexane several times, and sucking at room temperature for use to obtain oral fixed-release preparation for fixed-release in terminal ileum and colon, wherein the microsphere is shown in figure 6.

The prepared component A and component B are respectively prepared into the following 4 oral preparations:

the 1 st: preparing 6gA components into an oral preparation;

the 2 nd: preparing 6gB components into an oral preparation;

and (3) type: 3g of each of the component A and the component B are prepared into a mixed oral preparation according to the ratio of 1:1 (obviously, the mixing ratio can be adjusted).

And 4, the method comprises the following steps: the blank solution without microspheres was prepared as an oral formulation as a control.

8 overnight fasted insulin-independent type 2 diabetic patients were divided into 4 groups in the early morning, each of 2 patients was randomized into one group, and the patients in each group were administered the above 4 preparations, which were labeled as group 1, group 2, group 3, and group 4, respectively.

30 minutes after oral administration of the formulation, the patient received an oral glucose tolerance test. Blood was collected at the following time points: 30, 0, 15, 30, 60, 90, 120 minutes.

Blood is analyzed for the levels of hormones such as GLP-1, PYY, insulin, glucose, etc. Several of the indices were significantly improved in patients in groups 1, 2, and 3 compared to the oral placebo control group, with the greatest degree of improvement in group 3.

In addition, the invention also researches and adopts Chitosan-Catechol (Chitosan-Catechol, CC) as the intestinal mucosa adhesive polymer to prepare the Chitosan-Catechol microsphere. The results of testing the mucosa adhesiveness of the CC microsphere and the TC microsphere, respectively, are shown in fig. 9, where both the CC microsphere and the TC microsphere have good mucosa adhesiveness.

Therefore, referring to the preparation method of TC microspheres, Chitosan-Catechol microspheres are prepared using Chitosan-Catechol (Chitosan-cathechol, CC) as the intestinal mucosa adhesive polymer, and then oral duodenal targeting release preparations are prepared as the a component. Referring to the above experimental methods, the prepared a and B fractions were prepared into 4 oral formulations, respectively, and the results showed that several indices of the patients in groups 1, 2, and 3 were significantly improved compared to the control group orally administered with placebo, with the greatest degree of improvement in group 3.

The person skilled in the art will be able to identify and know that: the above example is merely one preferable embodiment known at present, and is not limited to the specific formulation forms listed in the examples, as long as it can combine:

1. an oral duodenal orientation release preparation with a shielding effect formed by the adhesion of intestinal mucosa adhesive material to the duodenal mucosa is used as a component A;

2. the B component can be oral fixed-release preparation with nutrient components released at ileum terminal and colon fixed-release.

While the invention has been described with respect to a preferred embodiment, it will be understood by those skilled in the art that the foregoing and other changes, omissions and deviations in the form and detail thereof may be made without departing from the scope of this invention. Those skilled in the art can make various changes, modifications and equivalent arrangements, which are equivalent to the embodiments of the present invention, without departing from the spirit and scope of the present invention, and which may be made by utilizing the techniques disclosed above; meanwhile, any changes, modifications and variations of the above-described embodiments, which are equivalent to those of the technical spirit of the present invention, are within the scope of the technical solution of the present invention.

Claims

1. A diabetic treatment product comprising an oral targeted release formulation and an oral duodenal targeted release formulation, wherein the oral targeted release formulation provides targeted release in the terminal ileum and colon, the oral targeted release formulation comprises a nutritional ingredient, the oral duodenal targeted release formulation comprises an intestinal mucosa adhesive polymer, the terminal ileum refers to the part of the ileum close to the colon, the intestinal mucosa adhesive polymer is selected from thiolated chitosan or chitosan-catechol, the nutritional ingredient is a combination of fatty acid and amino acid, and the amino acid is selected from arginine or glutamine.

2. The diabetes treatment product of claim 1, wherein the fatty acid is selected from the group consisting of: acetic acid, propionic acid, butyric acid, bile acid, and long chain fatty acid.

3. The diabetes treatment product of claim 1, wherein the nutritional ingredients are: a combination of propionic acid and arginine, a combination of propionic acid and glutamine, a combination of acetic acid and arginine, a combination of acetic acid and glutamine, a combination of butyric acid and arginine, a combination of butyric acid and glutamine, a combination of bile acid and arginine, a combination of bile acid and glutamine, a combination of long chain fatty acid and arginine, or a combination of long chain fatty acid and glutamine.

4. The diabetes treatment product of claim 3, wherein the mass ratio of the two nutritional ingredients in each combination is (1-10): (1-10).

5. The diabetes treatment product of claim 1, wherein the nutritional composition is encapsulated in an enteric mucoadhesive polymer.

6. The diabetes treatment product of claim 1, wherein the oral targeted release formulation for targeted release in the terminal ileum and colon is an enteric coated release formulation.

7. The diabetes treatment product according to any one of claims 1 to 6, wherein the diabetes treatment product is used for the treatment of type I diabetes, type II diabetes, pre-diabetes, obesity, appetite control, metabolic syndrome or polycystic ovary syndrome.

8. The product for the treatment of diabetes mellitus according to any one of claims 1 to 6, which is a substitute for metabolic surgery treatment.

9. Use of a diabetes treatment product according to any one of claims 1 to 6 in the manufacture of a medicament for the treatment of type I diabetes, type II diabetes, pre-diabetes, obesity, appetite control, metabolic syndrome or polycystic ovary syndrome.