WO2019037682A1 - Enteric oxygen microcapsule for adjusting intestinal flora, preparation method therefor and use thereof - Google Patents

Abstract

An enteric oxygen microcapsule for adjusting intestinal flora. The enteric oxygen microcapsule consists of an enteric capsule wall and a core material containing oxygen and/or an oxygen source. The oxygen in the core material accounts for 5-70% (V/V) of the total volume of the enteric oxygen microcapsule, and the oxygen concentration is 5-100%. The mass ratio of the core material to the enteric wall is 1:50-900. Also disclosed is a preparation method for the enteric oxygen capsule for adjusting intestinal flora, and further disclosed is an application of the enteric oxygen microcapsule in the preparation of drugs for treating diseases associated with intestinal flora imbalance such as obesity, diabetes, hypertension, cancer, or Alzheimer's or drugs for expelling ascarids, by using the enteric oxygen capsule as a sole active ingredient or one of a number of active ingredients.

Description

Enteric oxygen microcapsule for adjusting intestinal flora, preparation method and use thereof Technical field

The invention relates to the technical field of medicine, in particular to an oxygen microcapsule for regulating intestinal flora, a preparation method thereof and the use thereof as a pharmaceutical active ingredient for adjusting the structure of an intestinal flora in preparing a medicament.

Background technique

Oxygen is one of the most important substances in life activities.

Oxygen (O ₂ ), the most common elemental form of oxygen. Not easily soluble in water, accounting for about 21% in the air.

Oxygen is a double-edged sword. While maintaining the body's survival, it also produces a lot of free radicals.

Oxygen, the concentration in the environment, is a determining factor in regulating the distribution of microorganisms in the environment.

Oxygen is a broad-spectrum antibiotic-like effect and a special irreplaceable antibiotic-like substance. This is also the main mechanism of hyperbaric oxygen therapy (HBOT). Hyperbaric oxygen therapy (HBOT) is a treatment aid and rehabilitation method often used in hospitals to breathe oxygen in an environment of more than one atmosphere. One of its functions is to use the antibacterial effect of oxygen: oxygen is a broad-spectrum antibiotic, and high-concentration oxygen is not only resistant to anaerobic bacteria but also to aerobic bacteria.

But we know that regardless of the oxygen concentration in the environment, the oxygen carrying capacity of hemoglobin is generally relatively stable. In addition, in the calm state, the number of mesenteric capillaries is only about 1/5 - so normal people do hyperbaric oxygen therapy more than just comfort.

This is the main reason why hyperbaric oxygen therapy (HBOT) does not replace aerobic exercise - the amount of oxygen entering the intestine is still very low - unless dancing in the hyperbaric oxygen chamber jumps to fever.

In 2013, the US Food and Drug Administration (FDA) issued a warning on hyperbaric oxygen therapy in the consumer warning section of its website: Currently, there is no evidence that hyperbaric oxygen therapy is clinically effective in treating cancer, autism or diabetes. Or valid for it.

Microencapsulation is a technology in which trace substances are encapsulated in polymer films. It is a micro-packaging technology for storing solids, liquids and gases. The size is generally from 5 to 200 μm, and the shape is varied depending on the raw materials and the preparation method.

Among them, the embedded substance is called a core material, and includes flavors and fragrances, acidulants, sweeteners, colorants, lipids, enzymes, microorganisms, and gases, and various other additives. A substance that achieves microencapsulation by embedding a core material is called a wall material.

On June 27, 2012, the cover article of Science Translational Medicine, a science publication, reported reports from Dr. Kheir and colleagues from Boston Children's Hospital. They inject oxygen particles into the hypoxic animal to restore oxygen saturation to near normal levels in seconds. Chinese patent, "Particles, methods for their manufacture and applications in diagnosis" (CN1064851), the Abstract: The present invention relates to a contrast agent composed of gas-containing particles for ultrasonic diagnosis, the particle wall being composed of polycyanoacrylate Or a polyester of α-, β- or γ-hydroxycarboxylic acid.

Chinese patent, "Ultrasonic contrast agent and its manufacturing method and application in diagnostic treatment" (CN1033840) The invention relates to an ultrasonic contrast agent made of microparticles, which is composed of amylose or a biodegradable synthetic polymer and A gas and/or a liquid having a boiling point below 60 ° C.

Chinese patent CN102973594A discloses an ophthalmic preparation containing oxygen microcapsules and a preparation method thereof, and the oxygen microcapsules are mainly used for controlling various eye diseases caused by hypoxia.

Chinese patent CN1191836 discloses the use of hydrogen as a pharmaceutically active ingredient in the preparation of a pharmaceutical product. In addition, the hydrogen applied by the patenter Messer Group in the specification for the preparation of a pharmaceutical product describes that hydrogen is suitable for preparation. Treatment of inflammatory lesions in humans and mammals.

The human brain can use gas as a gasotransmitters, such as hydrogen sulfide, ammonia, or even carbon monoxide to transfer information between cells. Gut bacteria also produce gas neurotransmitters that affect our brains, moods and behaviors. (Oleskin, A.V. and B.A.Shenderov (2016). "Neuromodulatory effects and targets of the SCFAs and gasotransmitters produced by the human symbiotic microbiota." Microbial Ecology in Health and Disease 27:30971.)

In the human intestine, studies have shown that the volume of gas is less than 200 ml, generally 31 ml to 200 ml, with an average of 100 ml; individual differences are large.

The composition of the gas in the lumen varies with the specific location of the entire gastrointestinal tract. The gas in the stomach is similar to air, and the composition of the fart shows a huge individual difference. This reflects individuals with different constitutions and health conditions with different metabolic processes in the gastrointestinal tract.

More than 99% of the gas consists of five odorless gases: N ₂ , O ₂ , CO ₂ , H ₂ and CH _{4 ,} with a small amount of other odorous gases.

The ratio of N ₂ can be from 11% to 92%;

The ratio of O ₂ can be from 0% to 11%;

The ratio of CO ₂ can be from 3% to 54%;

The ratio of H ₂ can be from 0% to 86%;

The ratio of CH ₄ can range from 0% to 56%.

There are three sources of intestinal gas: air that is swallowed consciously or unconsciously, gases produced by bacteria in the gastrointestinal tract, and gases that diffuse from the blood.

Regarding the gas metabolism of the intestine, it can be summarized as two aspects of source and elimination. See Table 1.

Table 1 Gas metabolism in the intestine

Gut gas source

Swallowing gas (N ₂ , O ₂ )

Blood dispersion (N ₂ , O ₂ , CO ₂ )

· Hydrocarbonate neutralization (CO ₂ )

· Bacterial metabolism (H ₂ , CO ₂ , CH ₄ and trace gases).

Intestinal gas removal

·Helium

·Diffuse into the blood

·Bacterial metabolism

·Anal exhaust

In order to monitor the metabolism of gastrointestinal gases, it is beneficial to human health management. A team of professors at the Royal Melbourne Institute of Technology, Kourosh Kalantar-zadeh, developed a "smart capsule" for human intestinal gas monitoring. Its purpose is to measure intestinal gas samples in the most accurate and real-time field.

To determine the metabolism of intestinal microbes, gas samples are a good indicator of biology. Because different types of bacteria produce different gases, which in turn can affect intestinal function and changes in the intestinal flora.

Anaerobic bacteria, which are mainly composed of thick-walled bacteria, produce hydrogen, which can be further utilized by methanogenic archaea, sulfur-reducing bacteria or acetogenic bacteria to produce methane and hydrogen sulfide. (Ou, J. Z. et al. (2015) Human intestinal gas measurement systems: in vitro fermentation and gas capsules. Trends in Biotechnology, In Press.)

The gut microbiota, known as the "second genome" of humans, has become one of the most popular research fields in recent years.

These highly diverse and amazing numbers of bacteria live in our bodies.

Bacteria are extremely complex and are classified into aerobic, facultative anaerobic, anaerobic, and obligate anaerobic bacteria based on the difference in bacterial oxygen requirements.

In the intestines, various bacteria are combined according to a certain proportion, interdependent and mutually restrictive, forming a certain ecological balance in quality and quantity. When the internal or/and external environment changes, the normal ecological balance is broken, triggering an imbalance in the intestinal flora.

A number of recent studies have shown that diet, lifestyle, drugs, etc. will affect the type, quantity and function of intestinal flora, causing imbalance of intestinal flora, which makes the body's immune system have different responses. Changes in the intestinal flora and immune system promote insulin resistance and chronic inflammation. Obesity, diabetes, hypertension, cancer, Alzheimer's disease, neurodegenerative diseases, Parkinson's disease, etc. may appear.

The imbalance of intestinal flora is closely related to various diseases. (Gut microorganisms affect our physiology.Medical press.December 29, 2016.)

The relationship between intestinal flora imbalance and human host health is extremely complex, and the related pathophysiological mechanisms are far from clear.

The relationship between intestinal flora imbalance and obesity, diabetes and hypertension:

Metabolic diseases such as obesity and diabetes have become one of the most serious social and health problems in the world today. The current treatments are mainly for the consequences of disease rather than metabolic disorders, and it is difficult to curb the epidemic trend of global patients, so we need to find new and more effective prevention strategies. Obesity and diabetes occur as a result of genetic and environmental factors, but genomic analysis can only explain 10% to 20% of metabolic diseases. Environmental factors such as eating habits are very important in the pathogenesis of metabolic diseases. effect. In recent years, more and more studies have shown that human intestinal flora is closely related to the occurrence and development of metabolic diseases such as obesity and type 2 diabetes. The rapid advances in this emerging academic field will not only help elucidate the pathogenesis of metabolic diseases, but will also provide direction for exploring new therapeutic targets and pathways. (Research progress on the relationship between Gu Lien's intestinal flora and obesity and type 2 diabetes. Chinese Journal of Endocrinology and Metabolism 2015, 31(7): 641-645)

On March 20, 2013, JCEM: The breath test can determine whether a person has an obesity tendency.

This is the world's first large-scale study, confirming the link between ingredients and body weight in gas, which may help reveal another important factor in obesity. The researchers analyzed the gas contents exhaled by 792 people. Based on the breath test, the subjects showed four patterns:

1. Have normal gas contents;

2. There is a higher concentration of methane;

3. Have a higher level of hydrogen;

4. Both methane and hydrogen are high levels.

The high concentration of the two gases is closely related to a higher body mass index (BMI) and a higher percentage of body fat.

The presence of methane is associated with the intestinal methanobrevibacter smithii. This bacterium is responsible for the production of most of the methane in the human host.

Brevibacterium methane is an obligate anaerobic bacterium. It can be isolated in the gastrointestinal tract, anaerobic digester or other environment that is deficient in oxygen and has no other inorganic electron acceptors (such as nitrate or sulfate) other than CO ₂ .

Studies have shown that patients with periodontal disease have seven times the probability of developing diabetes.

Periodontal disease and diabetes have a two-way relationship. Diabetes is one of the risk factors for circumferential disease, and periodontal disease can also affect diabetes.

Studies have shown that the proportion of Gram-negative anaerobic bacteria in adults with periodontitis is about 75%.

It has been reported that diabetic patients with periodontitis have decreased glycated hemoglobin levels after periodontal treatment and reduced the amount of diabetes medication. ([1]. Mealey BL, Rose LF. Diabetes mellitus and inflammatory periodontal diseases. Compend Contin Educ Dent, 2008, 29(7): 402-408, 410, 412-403. [2]. Shang Yihuan and other different ages in Wuhan Correlation study on risk factors of periodontitis in civil servants. Chinese Journal of Geriatrics, 2008; 27(12): 931-934.

On August 21, 2014, Immunity reported that the intestinal bacterial metabolite Indole 3-propionic acid (IPA) may maintain the defense function of intestinal epithelial cells. (doi:10.1016/j.immuni.2014.06.014.Epub 2014Jul 24.)

A new study from the University of Eastern Finland published on Scientific Reports on April 11, 2017 showed that high levels of sputum in serum can prevent type 2 diabetes.

Propionic acid is a metabolite produced by intestinal bacteria, and the body can enhance the production of propionic acid through high-fiber foods. (Indolepropionic acid and novel lipid metabolites are associated with a lower risk of type 2diabetes in the Finnish Diabetes Prevention Study doi:10.1038/srep46337)

However, it is not clear at this time the specific source of flupropionic acid in the intestine.

We know that tryptophan is a derivative of guanidine that can be metabolically degraded by bacteria in the intestines. For example, tryptophan is metabolized to produce 5-hydroxy-3-indolyl-α-ketone.

Based on this, the best advice given by scientists is to enhance the production of cyanopropionic acid through high-fiber foods.

On October 29, 2015, a research article published on Cell Metabolism stated:

The short-chain fatty acids produced by the decomposition of intestinal bacteria increase the accumulation of lipids in the liver, which will cause the body to suffer from non-alcoholic fatty liver and liver damage. The researchers also found that mice that overdose dietary fiber had reduced Toll-like receptor 5 (TLR5) function and caused excessive bacterial growth in the gut. (Gut bacteria could be blamed for obesity and diabetes, MNT, 30, Oct, 2015)

Previous studies have shown that obesity is associated with cardiovascular disease. Recently, a British study published in the JAMA Cardiology Journal once again confirmed that overweight/obesity is associated with an increased risk of cardiac metabolic diseases, including coronary heart disease, hypertension, and type 2 diabetes. The study included nearly 120,000 people. (Lyall DM, Celis-Morales C, Ward J, et al. Association of Body Mass Index With Cardiometabolic Disease in the UK Biobank: A Mendelian Randomization Study. JAMA Cardiol. Published online July 5, 2017.)

Researchers already know that diet affects the balance of microbes in the body, but how this translates into a host's effects is not yet understood. Now, studies in mice have shown that microbes interact with the host by releasing metabolites that act on histones, thereby affecting gene transcription, not only in the colon, but also in tissues elsewhere in the body.

The relationship between intestinal flora imbalance and Alzheimer's disease:

Earlier, the University of California, Los Angeles (UCLA) study found that by feeding antibiotics, "keeping the balance of intestinal bacteria helps delay the disease associated with aging." Professor Walker said, "This study may bring more realistic methods for scientists to intervene in the aging process, delay Parkinson's disease, Alzheimer's disease, cancer, stroke, cardiovascular disease, diabetes, etc., even though these Progress may take many years." (Cell Reports, 2015, 12(10): 1656)

Antibiotics have also been shown to be resistant to Alzheimer's disease (AD). "Antibiotic-induced perturbations in microbial diversity during post-natal development alters amyloid pathology in an aged APPSWE/PS1ΔE9murine model of Alzheimer’s disease" (Scientific Reports volume 7, Article number: 10411 (2017))

In a December 2016 study published in Neurology, researchers from the Institute of Neurology and Brain Research at the University of California, Davis, first discovered that there were differences in brain samples from patients with delayed Alzheimer's disease. High levels of Gram-negative bacteria antigens in these brain samples have higher levels of bacterial lipopolysaccharide and E. coli K99 pilin.

In addition, the researchers also found that these lipopolysaccharide molecules can accumulate in amyloid plaques, which is directly related to the pathological manifestations and disease progression of patients with Alzheimer's disease.

The article said: This study reminds researchers to conduct more in-depth research to investigate the mechanism of infectious factors affecting Alzheimer's disease, to clarify the role of bacteria in their pathogenesis, bacterial infectious agents and their pathogenesis The interrelationship provides new ideas and new hopes for the development of new treatments.

(Xinhua Zhan, MD, Boryana Stamova, PhD, Lee-Way Jin, MD, Charles DeCarli, MD, Brett Phinney, PhD and Frank R. Sharp, MD. Gram-negative bacterial molecules associate with Alzheimer disease pathology. Neurology. 2016)

It can be seen that bacteria are the association of the above two research results on Alzheimer's disease. And we know that Gram-negative anaerobic bacteria account for 75% of adults with periodontitis.

The relationship between intestinal flora imbalance and cancer:

Hippocrates: Every disease begins in the intestines. Nobel laureate immunologist Pioneer Mechnikov: Death begins in the large intestine.

Aging is associated with increased levels of tumor necrosis factor (TNF) and pro-inflammatory cytokines in blood and tissues, which induce inflammation associated with aging. (Age-Associated Microbial Dysbiosis Promotes Intestinal Permeability, Systemic Inflammation, and Macrophage Dysfunction Cell Host & Microbe, VOLUME 21, ISSUE 4, P455-466.E4, APRIL 12, 2017)

Dawn Bowdish said: Currently, the only way to reduce inflammation associated with aging is to maintain a healthy diet and exercise, and to control as much as possible any chronic inflammatory disease. We hope to use drugs, probiotics or probiotics in the future to increase the barrier function of the intestines, leaving microbes to stay in their place where they live, thereby reducing inflammation associated with aging.

Research in the 2014 Lancet magazine revealed that the probability of cancer in overweight and obese people is significantly higher than that of the average person. The researchers counted 12,000 cancers caused by obesity each year in the UK, and the proportion of cancer in obese people is expected to increase at a rate of 3,700 per year. (Bhaskaran K, Douglas I, Forbes H, et al. Body-mass index and risk of 22 specific cancers: a population-based cohort study of 5·24million UK adults. The Lancet 13August 2014.)

In October 2011, two research groups from the Canadian BC Cancer Institute and the Broad Institute confirmed that F. nucleatum is also present in the intestine, and its abundance in the intestine and colorectal cancer Related.

Fusobacterium nucleatum is a Gram-negative obligate anaerobic bacterium that has the potential to cause disease and can cause periodontal disease.

Recent studies have shown that Fusobacterium nucleatum may promote colorectal cancer by interfering with the body's immune system and by activating the growth pathways of colon cells. (Mehta RS, Nishihara R, Cao Y et al. Association of Dietary Patterns With Risk of Colorectal Cancer Subtypes Classified by Fusobacterium Nucleatum in Tumor Tissue. JAMA Oncol. 2017 Jan 26.)

Obviously, Fusobacterium nucleatum, and the methane-producing Brevibacterium methanei described above in this specification, are obligate anaerobic bacteria, plus anaerobic bacteria that produce hydrogen as the main force of thick-walled bacteria. Oxygen bacteria are very happy to stay in a low-oxygen or even anaerobic environment of their own preference.

Taken together, various phenomena indicate that obesity, diabetes, especially type 2 diabetes, cancer and Alzheimer's disease are closely related to intestinal anaerobic or obligate anaerobic bacteria.

Aerobic exercise refers to physical exercise performed by the human body with sufficient oxygen supply. That is, during exercise, the oxygen inhaled by the human body is equal to the demand and reaches a physiological balance. In simple terms, aerobic exercise refers to any rhythmic exercise that has a longer exercise time (about 15 minutes or more) and a moderate or upper exercise intensity (75% to 80% of maximum heart rate). Aerobic exercise is a constant exercise, and it is exercise that lasts for more than 5 minutes. For example, walking, swimming, and cycling are all aerobics.

Professor Hu Dayi's speech in "Aerobic Metabolism Exercise and Cardiovascular Disease", using specific examples, personal experience, detailed the benefits of aerobic exercise. "Aerobic capacity" has been included in the "Clinical Vital Signs" in the United States (http://circ.ahajournals.org/content/134/24/e653).

Aerobic exercise helps lower blood pressure and is widely recommended by the US and European guidelines for hypertension. Aerobic exercise, even in an uncontrolled and unsupervised manner, can significantly reduce dynamics and office blood pressure. (Pagonas, Nikolaos; et al. Aerobic versus isometric handgrip exercise in hypertension: a randomized controlled trial. Journal of Hypertension. 2017.)

Recently, a study published in the journal Diabetes Care was designed to assess the effects of 20-week aerobic exercise training on left ventricular function and glycemic control in normal adolescents and type 1 diabetic adolescents. The conclusion is that aerobic exercise can reduce the insulin dosage by about 10% and improve the heart function of juvenile diabetes. (Gusso S, Pinto T, et al. Exercise Training Improves but Does Not Normalize Left Ventricular Systolic and Diastolic Function in Adolescents With Type 1Diabetes.Diabetes Care.2017Jul 18.pii:dc162347.doi:10.2337/dc16-2347.)

Mild cognitive impairment (MCI) is an intermediate state between normal aging and dementia. It occurs mostly in the elderly over 65 years of age and shows a slight decline in cognitive ability, but this symptom does not cause daily life. Significant negative impact. Studies have shown that about 5-20% of the elderly suffer from mild cognitive impairment. MCI often develops Alzheimer's disease (AD). Some published literature reveals that 80% of patients with mild cognitive impairment will progress to Alzheimer's disease after 6 years. Previous research suggests that aerobic exercise is expected to increase the volume of specific brain regions, thereby increasing memory. Recent research confirms that exercise not only increases brain volume, but also improves cognitive performance in patients with MCI (Aerobic exercise improves cognition in old age. Published Wednesday 30 November 2016, http://www.medicalnewstoday.com/articles/314448. Php).

Although aerobic exercise is very beneficial to the body, doctors often warn that it is not greedy, excessive exercise will produce a lot of free radicals, etc., which will have a negative effect. Therefore, aerobic exercise is a degree, and this degree is currently controlled by time and heart rate.

In 2014, Nobel Laureate, one of the discoverers of DNA structure, James Dewey Watson, presented his thoughts on diabetes in a cover article published in The Lancet: Watson believes that type 2 diabetes The pathogenesis is not oxidative stress, but rather because of insufficient oxidative stress in cells; and he extends the theory of oxidative stress to Alzheimer's disease and cancer. (Watson J.D. Type 2daibetes as a redox disease [J]Lancet, 2014, 383 (9919): 841-843.)

Watson’s speculation stems from his 2013 study of Michael Ristow and his colleagues in 2009:

The sensitivity of insulin and the amount of reactive oxygen species in the two groups of men after physical exercise were compared. One group took vitamin C and vitamin E, and the other group did not.

It was found that the amount of active oxygen in the group without vitamins increased, and the amount of active oxygen increased was directly proportional to the sensitivity of insulin after exercise, while the amount of active oxygen in the group taking vitamin C and vitamin E increased. It does not change, and the sensitivity of insulin does not increase after exercise.

And many other tests on antioxidants confirm the findings: almost all trials that used vitamin C and vitamin E to improve athletic performance failed.

The inventor believes that this interesting phenomenon that causes Watson's attention can be explained in this way:

1. Generally speaking, aerobic exercise, the body cells of the body need more oxygen, breathing faster, blood circulation is intensified, and oxygen from the blood diffuses into the intestines. The inventors believe that this is the most fundamental mechanism for aerobic exercise in health benefits - to increase the oxygen content of the intestinal environment of the body, to inhibit the anaerobic activity, and to regulate intestinal flora.

2. Vitamin C and vitamin E are extremely susceptible to oxidation. After entering the intestine, some of the vitamins are quickly oxidized by the oxygen in the intestines. In particular, fat-soluble vitamin E requires a certain amount of fat to prevent it from being rapidly oxidized. Vitamin E is absorbed in the form of micelles in the middle of the small intestine. Its absorption is similar to the absorption of dietary fat, and bile must be present. The absorption rate under normal conditions is 20% to 25%.

As a result, the increased oxygen content in the intestine due to aerobic exercise is also aggravated by the oxidation of vitamins C and E.

According to the principles of evidence-based medicine, we also saw an interesting phenomenon associated with it.

On July 21, 2015, Nature magazine reported:

Draining bile acids into the last part of the small intestine is sufficient to produce a weight loss similar to more complex conventional bariatric surgery.

Researchers believe that reduced fat absorption in the small intestine and changes in the intestinal flora are part of the reason for these effects. (Bile diversion to the distal small intestine has comparable metabolic benefits to bariatric surgery. Nature Communications 6, Article number: 7715 (2015))

Or it may be suggested that vitamin E reduces the reaction with intestinal oxygen due to the action of fat, and oxygen is not reduced by vitamin E. Conversely, the oxygen content of the intestine may also be increased during the operation of the procedure.

The inventor has reason to believe that in the near future, for the monitoring of aerobic exercise, a monitoring index of "intestinal oxygen amount" will be added, which is regarded as the main inventor of the "intelligent capsule" of the intestinal gas monitoring. Professor Rosh’s wish.

In general, the intestines are usually deficient in oxygen.

Based on this, the "bio-oxygenation" mechanism for adjusting the balance of intestinal flora has emerged.

"Intestinal Health", a live Bacillus licheniformis preparation. The main mechanism of action is to treat bacteria by bacteria, resulting in a hypoxic environment in the intestine. However, this is only the tip of the iceberg that uncovers the imbalance of intestinal flora, and it is far from the imbalance of the intestinal flora.

It goes without saying that there is no absolute balance in the world. Everything is in dynamic balance. There is no balance that can be perfect, so that all aspects benefit and are satisfied. The same is true for the balance of the intestinal flora.

The inventors believe that the determination of whether the intestinal flora is unbalanced should be based on the "presumption of innocence". Rather than saying “intestinal flora imbalance”, it is necessary to supplement “probiotics”.

We know that even dietary fiber is not much better, and the inventors do not believe that probiotics are safer than dietary fiber.

Whether the intestinal flora is out of balance depends on changing the environment inside or/and outside the host, especially in the host's intestinal environment, whether it can bring health benefits to the host.

And artificially divide the bacteria's good or bad, label the "good bacteria" and "bad bacteria", and use "good bacteria" or / and "bad bacteria" to determine the imbalance of intestinal flora or not. If there is a misconduct, it can be said to be arbitrary and unscientific.

There is a common sense: power is always in the hands of a few. Bacteria also have hands, just non-hands.

The original atmosphere is oxygen free.

At a macro level, all diseases may be struggles or efforts by anaerobic or anaerobic organisms to survive or rebuild their homes.

For major diseases, especially some so-called "difficult diseases", it is a good idea or idea to give anaerobic bacteria a strong focus.

In addition, if the intestinal flora imbalance can be applied to the "bio-oxygenation" mechanism, then those living bacteria preparations that meet the "bio-oxygenation" mechanism are not to cure all diseases, and should be more than one type. Several diseases will have a good therapeutic effect.

However, this is not the case.

It can be said that blindly applying the "biological oxygen scavenging" mechanism, the hard-working "intestinal flora imbalance" may bring many adverse effects to the body, and the risk is not small.

The intestinal flora is extremely complex and is currently poorly understood. Investigation and research on the substrate or metabolic products degraded by bacteria is more conducive to discovering the clues of some diseases related to the imbalance of intestinal flora.

Phenylalanine, an essential amino acid. Any natural protein contains about 4% phenylalanine. (The content of animal protein is more than that of vegetable protein. Another thoughtful phenomenon about phenylalanine.)

Phenylalanine, like tryptophan, is also an aromatic amino acid.

Bacteria having phenylalanine deaminase activity can deamination of phenylalanine in the medium to phenylpyruvate, which turns the ferric chloride indicator into green. Proteus, Providencia and Morganella were positive, while other bacteria in Enterobacter were negative.

Proteus, Providencia, and Morganella are both facultative anaerobic bacteria. Among them, the anaerobicity of the genus Providencia and the genus Mortierella is even worse than that of the genus Bacillus, and it also metabolizes tryptophan, which is metabolized by deamination to produce 5-hydroxy-3-propionic acid. Alpha-ketone, not guanic acid which is beneficial for diabetes.

Recently, on August 3, 2017, "Science" published a team of Professor Marco Colonna of the University of Washington School of Medicine to find the intestinal microbe that regulates specific immune cells, Lactobacillus reuteri. However, this Lactobacillus, which is almost present in the gut of all vertebrates and mammals, requires a tryptophan (Trp) to function. [Cervantes-Barragan L, Chai J N, Tianero M D, et al. Lactobacillus reuteri induces gut intraepithelial CD4+CD8αα+T cells[J].Science,2017:eaah5825.]

Lactobacillus reuteri is a facultative anaerobic bacterium, not an anaerobic bacterium, nor an obligate anaerobic bacterium.

In 2003, the inventors found that excessive phenylalanine in the diet is an important factor affecting the body's tryptophan metabolism. It was initially confirmed that reducing phenylalanine in the diet is beneficial to diabetic rats.

The idea of "a reasonable diet and a balanced Phe/Trp ratio in the diet is proposed."

Because insulin promotes the entry of amino acids into cells and reduces the concentration of amino acids in the bloodstream, it does not have this effect on tryptophan, which is not its amino acid component.

Tryptophan is the only amino acid that binds to plasma albumin and has both bound and free forms in plasma. Under normal circumstances, these two forms of tryptophan are in equilibrium. The bound type tryptophan is about 90% in plasma and about 10% in free form. Only free-type tryptophan can be directly used by tissues and can pass the blood-brain barrier. Insufficient intake of tryptophan will inhibit the production of antibodies in the body, causing abnormalities in immune regulation.

According to current research, only about 20% of human bacteria have been cultured and studied, and the other 80% of bacterial species are still unknown. Therefore, it is a long way to go to recognize these bacteria and master the laws of their activities.

The present inventors prepared a microbial preparation by using a natural non-pathogenic strain having phenylalanine deaminase activity and capable of metabolizing the properties of phenylalanine. Its preliminary pharmacodynamic experiments show a gratifying potential.

The inventors have never given up on further optimization work for many years. During this period, it was exciting to find that one strain was able to degrade tryptophan, but its metabolites were not the same as those of anaerobic fungi, the provenances of Morrowella and Morganella. Moreover, a characteristic of the strain in the presence of oxygen gives the inventors great touch.

It should be possible to create a new path to adjust the structure of the intestinal flora to re-construct a new balance in a model that is conducive to the health of the current body.

Increasing the oxygen content of the gut environment of mammals, especially human hosts, is a viable path.

Up to now, using microcapsule preparation technology, oxygen as core material, wrapped in enteric wall material, oxygen is sent to the intestines before release, in order to improve the intestinal environment oxygen content, adjust the intestinal flora, and then arrive The purpose of preventing or/and treating diseases associated with intestinal flora imbalance has not been reported in related literature and patents. Accordingly, there is an urgent need to provide a method for the use of oxygen microcapsules for regulating intestinal flora, methods for their preparation, and their use in the manufacture of medicaments for preventing or/and treating diseases associated with intestinal flora imbalance.

Summary of the invention

The term "intestinal flora imbalance" in the present invention means that the obligate anaerobic or/and anaerobic or/and facultative anaerobic bacteria in the intestinal tract of a host, particularly a human host, can be administered to give the host a health benefit.

The term "enteric solubility" in the present invention is not limited to the definition of conventionality, but should be understood as: "all feasible methods and tools for feeding oxygen into the intestine for the purpose of adjusting the intestinal flora".

The term "oxygen source" as used in the present invention means: a substance capable of generating oxygen, including the photosynthetic bacteria described herein before. Oxygen refers to conventional medical oxygen, and the term oxygen (O) has a conventional meaning and includes all isotopes of oxygen, ie, isotopes 16O, 17O or 18O. Oxygen molecules include a combination of all possible possibilities of isotopic oxygen with each other. The role of oxygen is not limited to the gas phase. A certain proportion of oxygen is completely dissolved in the liquid according to Henry's law.

The concentration of oxygen is too high, and the preparation is easily oxidized, so oxygen is mixed with an inert gas or other gas. The term "oxygen-containing gas mixture" also includes pure oxygen.

The technical problem to be solved by the present invention is to overcome the deficiencies of the prior art, to provide an oxygen microcapsule for adjusting intestinal flora, and an oxygen or oxygen source in the enteric oxygen microcapsule as an adjusted intestinal flora. Use of the sole active ingredient or one of the active ingredients in the preparation of a medicament for preventing or/and treating a disease associated with intestinal flora imbalance, or for the treatment of aphid, including prevention or/and treatment of obesity, diabetes, hypertension The use in the medicament for cancer and Alzheimer's disease also provides a preparation method for preparing the drug, i.e., enteric oxygen microcapsules, for achieving the use.

The pharmaceutical product of the invention may be in a variety of different forms. The pharmaceutical product may be a gas mixture or liquid containing an oxygen/oxygen source (eg, a gas mixture containing oxygen or pure oxygen dispersed in a fat emulsion) or a solid formulation (eg, the gas is in a clathrate).

The technical solution adopted by the present invention to solve the technical problem thereof is:

An enteric oxygen microcapsule for adjusting intestinal flora of the present invention, characterized in that the enteric oxygen microcapsule is made of an enteric capsule wall and an oxygen-containing core material, and oxygen in the core material The content is 75 to 100% (W/W), and the mass ratio of the core material to the wall of the capsule is 1:50 to 900, preferably 1:50 to 500, and most preferably 1:50 to 100.

Further, the enteric oxygen microcapsules are enteric microcapsules, enteric microparticles, enteric microdroplets, enteric microcapsule powder or enteric gel.

Further, oxygen includes one or more isotopes of oxygen, 16O, 17O or 18O.

In a certain exemplary embodiment, the core material comprises 1 to 15 parts of oxygen or/and oxygen source, wherein the oxygen concentration is 5 to 100%, so that the core material accounts for 5 to 70% of the total volume of the enteric oxygen microcapsule; The enteric wall includes 15-25 parts of sodium alginate, 15-25 parts of chitosan, 5-10 parts of polyvinyl alcohol, 35-65 parts of yeast powder and a small amount of cross-linking agent (eg, cross-linking agent The weight of the dialdehyde is 0.1 to 0.5% (w/w) of the enteric oxygen microcapsules.

The crosslinking agent is a combination of one or more of glutaraldehyde, CaCl2, BaCl2, FeCl3, CrCl3, genipin, an aqueous solution of ethylene glycol dimethacrylate, and sodium tripolyphosphate (TPP); preferably, pentane Dialdehyde acts as a crosslinking agent.

In an exemplary embodiment, the enteric oxygen microcapsules are enteric microcapsules: 1 to 100 parts of oxygen or/and oxygen source, and the oxygen concentration is 5 to 100%, so that the core material accounts for the total amount of enteric oxygen microcapsules. 5 to 70% by volume; the enteric wall is mainly made of the following raw materials: 10 to 300 parts of enteric polymer, 5 to 400 parts of surfactant, 1 to 40 parts of isotonicity adjusting agent, pH adjustment The amount of the agent is adjusted to the pH of the enteric microcapsule preparation to 6.6 to 8.7, and the balance is water.

Wherein, the enteric capsule wall further comprises an auxiliary agent such as a plasticizer, titanium dioxide or lake. The plasticizer is used in an amount of 0-35%, preferably 10-30%, of the enteric polymer, and the plasticizer includes but is not limited to triethyl citrate, glycerol acetate, glycerin, PEG6000, PEG4000, Tween 80, 1 2-propanediol, monostearic acid glyceride, preferably triethyl citrate. The amount of titanium dioxide or other lake is 2-40% by mass of the enteric polymer, preferably 10-30%, and most preferably 15-20%.

Enteric polymers are enteric materials commonly used in the field, including but not limited to formaldehyde gelatin, shell gum, cellulose acetate phthalate (CAP), copolymers of acrylic acid and methacrylic acid, methacrylic acid-A Methyl methacrylate copolymer, methacrylic acid-methyl acrylate (or ethyl ester, butyl ester) copolymer, ethyl acrylate-methacrylate copolymer, cellulose acetate, ethyl cellulose, polyacrylic resin, acrylic acid Ethyl ester-methyl methacrylate (2:1) copolymer, silicone elastomer (silicone rubber), polyvinyl acetate phthalate (PVAP), hypromellose phthalate (HPMCP) , hydroxypropyl methylcellulose peptidate, hydroxypropylmethylcellulose succinate (HPMCAS) or cross-linked alginate.

The surfactant is mainly a phospholipid, and the phospholipids include, but are not limited to, natural soybean phospholipids, natural lecithin, or derivatives thereof, and the mass ratio of oxygen to phospholipid is 1:0.1-100. The optimum is 1:10-50.

The enteric oxygen microcapsules of the present invention for regulating the intestinal flora with oxygen as the sole active ingredient or active ingredient can also be used for the preparation of a drug containing a lipid-containing, peptide-containing or protein-containing liquid.

The preparation method of the enteric oxygen microcapsule for adjusting the intestinal flora of the invention comprises the following steps:

1) preparing an aqueous solution of an emulsifier, a thickener, and a stabilizer, respectively;

2) preparing an enteric cyst wall aqueous solution;

3) The oxygen in the core material is passed through an orifice in the aqueous solution of the emulsifier prepared in the step 1) under high pressure. After 1 hour, the chitosan and polyvinyl alcohol aqueous solution in the dissolved enteric wall are slowly added. Stir for 15min, then add yeast leachate aqueous solution, continue to stir and emulsify, then slowly add aqueous sodium alginate solution in the enteric wall, ice bath and stir;

4) adding a glutaraldehyde cross-linking agent dilution solution while stirring in an ice bath, and after the Weissenberg effect occurs, adjust the pH of the solution to disperse the solid into microcapsules, and then continue stirring for more than 10 minutes;

5) sifting the solution prepared above to remove large particles of impurities and precipitates; centrifuging the supernatant, washing with 5% sodium borohydride solution, and drying by suction filtration;

Wherein, the core material comprises 1 to 15 parts of oxygen or/and oxygen source, wherein the oxygen concentration is 5 to 100%, so that the core material accounts for 5 to 70% of the total volume of the enteric oxygen microcapsule; the enteric wall Including 15 to 25 parts of sodium alginate, 15 to 25 parts of chitosan, 5 to 10 parts of polyvinyl alcohol, 35 to 65 parts of yeast powder and a small amount of crosslinking agent. The preparation method of the enteric oxygen microcapsule for adjusting intestinal flora of the invention comprises the following steps:

1) soaking the enteric polymer to prepare an enteric coating solution;

2) dissolving the isotonic adjusting agent and the surfactant in the auxiliary material with water or adding a co-solvent to uniformly disperse;

3) combine the enteric coating liquid in step 1) and the auxiliary agent in step 2), add water or enteric polymer cosolvent to the full amount, stir at high speed, adjust the pH to 6.6 ~ 8.7;

4) passing oxygen into the high-speed stirring solution of step 3) to form fine microcapsules containing a large amount of oxygen, and homogenizing into stable oxygen microcapsules;

Wherein, the enteric oxygen microcapsule is an enteric microcapsule: 1 to 100 parts of oxygen or/and oxygen source, and the oxygen concentration is 5 to 100%, so that the core material accounts for 5 to 70% of the total volume of the enteric oxygen microcapsule. The enteric capsule wall is mainly prepared by using the following raw materials: 10 to 300 parts of enteric polymer, 5 to 400 parts of surfactant, 1 to 40 parts of isotonicity adjusting agent, and the amount of pH adjusting agent is regulating intestinal The microcapsule preparation has a pH of 6.6 to 8.7 and the balance is water.

The use of an enteric oxygen microcapsule for adjusting intestinal flora of the present invention is applied to the preparation of a medicament for regulating enteric oxygen microcapsules as one of the sole active ingredients or active ingredients for regulating intestinal flora.

Preferably, the enteric oxygen microcapsules are used in the preparation of a medicament for preventing or/and treating a disease associated with intestinal flora imbalance or a medicament for aphid.

Further, the disease associated with intestinal flora imbalance is one or more of obesity, diabetes, hypertension, cancer or Alzheimer's disease.

The invention has the beneficial effects of adjusting the enteric oxygen microcapsules of the intestinal flora: the composition is simple, the production cost is low, the preparation process is simple, and the regulation effect on the intestinal flora is good, especially for obesity, diabetes, Diseases related to intestinal flora imbalance such as hypertension, cancer or Alzheimer's disease have a good preventive/therapeutic effect and can also drive mites.

The enteric oxygen microcapsules in the form of enteric solid particles have good stability, are convenient for storage and transportation, and are beneficial for the oxygen/oxygen source to efficiently regulate the intestinal flora in the intestine, so that the intestinal flora reaches equilibrium.

The invention has the beneficial effects of preparing the enteric oxygen microcapsule of the intestinal flora: the operation is simple, the preparation can be performed by using the existing pharmaceutical processing equipment, no additional equipment is required to be purchased, the equipment investment cost is low, and the scale is convenient. Production.

Detailed ways

The invention is further illustrated by the following examples.

Example 1

The following technical solutions and operation steps are implemented:

An enteric oxygen microcapsule (referred to as NDDK1) for adjusting an intestinal flora, the enteric oxygen microcapsule being made of an enteric capsule wall and an oxygen-containing core material, wherein the enteric oxygen microcapsule is an intestine Solvent microcapsules, the specific composition is as follows: 20% oxygen 600ml; poloxamer 15.0g; soybean phospholipid 11.0g; glycerol 7.5g; cellulose acetate cellulose 4.0g; sodium hydroxide solution to adjust the pH value of 6.8; Add 1000 ml of water.

The preparation method of the enteric oxygen microcapsule for adjusting the intestinal flora of the embodiment comprises the following steps:

1) soaking the enteric polymer to prepare an enteric coating solution;

2) dissolving the isotonic regulator glycerin, the surfactant poloxamer with water or adding a co-solvent to uniformly disperse;

3) Combine the enteric coating liquid in step 1) and the solution in step 2), adjust the pH to 6.6-8.7 with NaOH solution, and then make up the water to the full amount (1000 ml), and stir at high speed;

4) The oxygen is introduced into the high-speed stirring solution of step 3) to form fine microcapsules containing a large amount of oxygen, and homogenized to form stable oxygen microcapsules by a homogenizer.

Among them, experimental drugs, instruments and reagents: oxygen is medical oxygen; instruments are homogenizers; excipients are all pharmaceutical excipients.

Calculation of the total volume of oxygen: The volume of the formulation is increased by the addition of oxygen.

Example 2

An enteric oxygen microcapsule (referred to as NDDK2) for adjusting an intestinal flora, the enteric oxygen microcapsule being made of an enteric capsule wall and an oxygen-containing core material, wherein the enteric oxygen microcapsule is an intestine Microcapsules, specific composition such as: 45% oxygen 300ml, poloxamer 13.0g, soybean phospholipid 10.0g, sodium chloride 4.0g, polyvinyl acetate phthalate 7.0g, sodium hydroxide solution to adjust pH The value is 7.6 and 1000 ml is added with water.

The preparation method of the enteric oxygen microcapsule for adjusting the intestinal flora of the present embodiment is the same as that of the first embodiment.

Example 3

An enteric oxygen microcapsule (referred to as NDDK3) for adjusting an intestinal flora, the enteric oxygen microcapsule being made of an enteric capsule wall and an oxygen-containing core material, wherein the enteric oxygen microcapsule is an intestine Microcapsules, specific composition such as: 90% oxygen 700ml, poloxamer 17.0g, lecithin 11.0g, glycerol 7.0g, shellac 9.0g, sodium hydroxide solution to adjust the pH value of 8.1, add water to make 1000ml .

The preparation method of the enteric oxygen microcapsule for adjusting the intestinal flora of the present embodiment is the same as that of the first embodiment.

Application examples and effects comparison

1) Effects of enteric oxygen microcapsules used to regulate intestinal flora on ob/ob obese mice

Experimental protocol: ob/ob mice, 50 males, 10 weeks old, weighing 50-55 g were randomly divided into 5 groups, namely, model group, NDDK1, NDDK2, NDDK3, correspondingly administered low, medium and high groups and normal. Group (control). Ten normal ob/ob mice were taken from each group. The model group, NDDK1, NDDK2, and NDDK3 were administered with low, medium, and high doses for 4 weeks. The rats were weighed and calculated for food intake, and glycosylated hemoglobin was measured. HbA1c), blood glucose, high density lipoprotein (HDL), low density lipoprotein (LDL) and triglycerides (TG). The differences between the respective drug-administered groups and the model group were compared. A T test was performed between groups. See Table 2 and Table 3 for the results.

Table 2 Effect of NDDK administration for 4 weeks on ob/ob mice (n=10)

*, P < 0.05, **, P < 0.01, compared with the model group

Table 3 Effect of NDDK administration for 4 weeks on blood biochemical parameters of ob/ob mice (n=10)

*, p<0.05, **, p<0.01, compared with the model group

It can be seen from Table 2 and Table 3 that the intestinal oxygen microcapsules used to adjust the intestinal flora have weight, glycated hemoglobin (HbA1c), blood glucose, and high-density lipoprotein (HDL) in obese mice regardless of the oxygen content. Low-density lipoprotein (LDL) and triglyceride (TG) have a good regulatory effect, indicating that the enteric oxygen microcapsules of Examples 1 to 3 have a preventive or/and therapeutic effect on diseases such as obesity and diabetes, that is, Enteric oxygen microcapsules for regulating intestinal flora can be used to prepare drugs for preventing or/and treating obesity and diabetes.

2) Effect of enteric oxygen microcapsules used to regulate intestinal flora on tumor growth of S180 tumor-bearing mice

Test animals: Kunming mice, weighing 18-22g, male and female, a total of 60, randomly divided into 5 groups, 12 in each group;

Tumor species: S ₁₈₀ tumor-bearing mice ascites;

Blank control group: distilled water;

Positive control group: cyclophosphamide injection;

Drug group: NDDK1, NDDK2 and NDDK3 according to Embodiments 1 to 3 of the present invention, that is, the oxygen content is divided into three doses of high, medium and low;

Experimental method: 60 Kunming mice were taken, and S ₁₈₀ tumor fluid (1×10 ⁷ /ml, 0.2 ml/only) was inoculated into the right axilla. After inoculation, they were randomly divided into the next day, and then NDDK was administered intragastrically and intraperitoneally. Cyclophosphamide (CTX), the dosage is calculated according to the body weight, and the blank control group is given the same volume of distilled water; the drug is administered for 10 days, weighed daily, and the neck is weighed after the drug is weighed the next day, and the subcutaneous tumor is removed. The block, called the tumor weight, calculates the tumor inhibition rate, and its calculation formula is:

The tumor inhibition rate (%) = (the average tumor weight of the control group - the average tumor weight of the experimental group) / the average tumor weight of the control group × 100,

The results are shown in Table 4.

Table 4 Effect of NDDK on tumor growth of S ₁₈₀ tumor-bearing mice

**Compared with blank group: P<0.01.

It can be seen from Table 4 that the enteric oxygen microcapsules used to adjust the intestinal flora have obvious inhibitory effects on the S ₁₈₀ tumor-bearing tumor regardless of the oxygen content, indicating the enteric oxygen microcapsule pairs of Examples 1 to 3. Diseases such as cancer have a preventive or/and therapeutic effect, that is, enteric oxygen microcapsules for adjusting the intestinal flora can be used for the preparation of a medicament for preventing or/and treating cancer.

Example 4

An enteric oxygen microcapsule for adjusting an intestinal flora, the enteric oxygen microcapsule being made of an enteric capsule wall and an oxygen-containing core material, the core material comprising oxygen or/and oxygen source 1~ 15 parts, wherein the oxygen concentration is 5 to 100%, so that the core material accounts for 5 to 70% of the total volume of the enteric oxygen microcapsule; the enteric wall includes 15 to 25 parts of sodium alginate, and chitosan 15 to 25 Parts, 5-10 parts of polyvinyl alcohol, 35-65 parts of yeast dipping powder and a trace amount of glutaraldehyde cross-linking agent.

Specifically, an enteric oxygen microcapsule for adjusting intestinal flora is prepared from the following materials: core material: oxygen, orifice diameter: 1.2 μm, pressure: 3200 Pa, continuous ventilation for 1 h; The soluble capsule wall includes 2.4 g of sodium alginate, 2.4 g of chitosan, 0.4 g of polyvinyl alcohol, 0.6 g of emulsifier Span 60, 11 g of yeast extract, and a crosslinking agent: glutaraldehyde, 4 ml.

The preparation method of the enteric oxygen microcapsule for adjusting the intestinal flora of the invention comprises the following steps:

1) Prepare an aqueous solution of emulsifier Span 60 and yeast extract powder separately: 0.6g Span 60 is dissolved in 60 ° C, 80 ml of distilled water, 11 g of yeast dipping powder is dissolved in 80 ml of distilled water, and yeast dipping powder is routinely treated: Live, rest, sieve,

2) Preparation of an aqueous solution of enteric cyst wall: 2.4 g of sodium alginate was dissolved in 65 ° C, 160 ml of distilled water;

2.4 g of chitosan was dissolved in 3 ml of 36% acetic acid solution diluted with 100 ml of distilled water at 65 ° C, and 12 g of polyvinyl alcohol was dissolved in 2000 ml of distilled water at 95 ° C;

3) The oxygen in the core material is passed through an orifice in the aqueous solution of the emulsifier prepared in the step 1) under high pressure. After 1 hour, the chitosan and polyvinyl alcohol aqueous solution in the dissolved enteric wall are slowly added. Stir for 15min, then add yeast leachate aqueous solution, continue to stir and emulsify, then slowly add aqueous sodium alginate solution in the enteric wall, ice bath and stir;

4) Add the glutaraldehyde cross-linking agent dilution while stirring in an ice bath. When the Weissenberg effect occurs, adjust the pH of the solution to 8.0. After 30 minutes, the solid is dispersed into microcapsules and stirring is continued. 20 minutes;

5) The solution prepared above was sieved to remove large particles of impurities and precipitates; the supernatant was centrifuged, washed with a 5% sodium borohydride solution, and dried by suction filtration to obtain a solid enteric oxygen microcapsule.

1) Effect of enteric oxygen microcapsules used to regulate intestinal flora on serum inflammatory factors in obese rats

Experimental protocol: healthy males, 55 obese rats (SD rats) of 4 weeks old, weighing 116±15.3g, feeding environment: maintaining indoor temperature 25±5°C, relative humidity 50%～60%, illumination 12h/d, Well ventilated; free drinking water, standard rat feed, for 3 days.

Ten rats were randomly selected as normal control group and fed with normal feed; the remaining 45 were used as obesity model, fed with high fat diet, and the body weight of the rats was weighed every Friday to measure the body length of the rats. Rats weighing more than 20% of the average body weight of the normal control group were taken 4 weeks later as a successful model rat.

Thirty rats with successful model were randomly divided into 3 groups and fed with normal feed, 25 g/d per rat.

The specific grouping is as follows:

Normal group (N=10): free drinking water, quantitative ordinary feed + appropriate amount of broken microcapsule powder;

Obese group (N=10): free drinking water, quantitative ordinary feed + appropriate amount of broken microcapsule powder (described in Example 4);

High dose group (N=10): free drinking water, quantitative common feed + 300mg/d microcapsule powder (described in Example 4);

Low dose group (N=10): free drinking water, quantitative normal feed + 100 mg/d microcapsule powder (described in Example 4).

The above 4 groups were divided into two cages, each with 5 cages. Ordinary feed, 250g/d per group, 125g/d per cage, once/d. Free drinking water. The enteric oxygen microcapsules for adjusting the intestinal flora described in Example 4 were separately mixed in the feed at a set dose.

After 2 weeks of intervention, the animals were sacrificed and relevant indicators were detected: including IL-1, IL-6, TNF-α and NF-κB. As described in Table 5.

Table 5 shows the changes of serum inflammatory factors in obese rats by adjusting enteric oxygen microcapsules of intestinal flora

Note: ###P<0.001, compared with the normal control group; *P<0.05, ***P<0.001, compared with the obese control group.

As can be seen from Table 5, the results show that the enteric oxygen microcapsules used to adjust the intestinal flora, and the enteric oxygen microcapsules (obese group) used after breaking the wall have no inhibitory effect on the serum inflammatory level of obese rats, and Compared with the obese control group, the directly used enteric oxygen microcapsules (described in Example 4, solid enteric oxygen microcapsules) were treated with serum IL-1, IL- in the obese rats after 2 weeks of intervention. 6. The levels of TNF-α and NF-κB decreased significantly (P<0.001), indicating that enteric oxygen microcapsules can significantly reduce the inflammatory response in obese rats, and the intestinal tract is increased with the amount of enteric oxygen microcapsules. The inflammatory slowing effect is more significant, indicating that the enteric oxygen microcapsules have the function of protecting the gastrointestinal mucosal barrier and can effectively promote the recovery of intestinal flora imbalance.

Claims (10)

1. An enteric oxygen microcapsule for adjusting an intestinal flora, wherein the enteric oxygen microcapsule is composed of an enteric capsule wall and a core material containing oxygen or/and an oxygen source, wherein the core material Oxygen accounts for 5 to 70% (V/V) of the total volume of the enteric oxygen microcapsules, and the oxygen concentration is 5 to 100%; the mass ratio of the core material to the enteric capsule wall is 1:50-900.
2. The enteric oxygen microcapsule for regulating intestinal flora according to claim 1, wherein the enteric oxygen microcapsule is enteric microcapsule, enteric microparticle, enteric microdroplet, enteric microcapsule Powder or enteric gel.
3. An enteric oxygen microcapsule for regulating intestinal flora according to claim 1 or 2, wherein the oxygen comprises one or more isotopes of oxygen, 16O, 17O or 18O.
4. The enteric oxygen microcapsule for regulating intestinal flora according to claim 2, wherein the enteric oxygen microcapsule is an enteric solid microparticle mainly prepared by using the following raw materials by weight: the core material Including 1 to 15 parts of oxygen or/and oxygen source, wherein the oxygen concentration is 5 to 100%, so that the core material accounts for 5 to 70% of the total volume of the enteric oxygen microcapsule; the enteric wall includes sodium alginate 15~ 25 parts, 15 to 25 parts of chitosan, 5 to 10 parts of polyvinyl alcohol, 35 to 65 parts of yeast powder and a small amount of crosslinking agent.
5. The enteric oxygen microcapsule for regulating intestinal flora according to claim 2, wherein the enteric oxygen microcapsule is an enteric microcapsule: 1 to 100 parts of an oxygen or/and oxygen source, and an oxygen concentration 5 to 100%, the core material accounts for 5 to 70% of the total volume of the enteric oxygen microcapsule; the enteric capsule wall is mainly made of the following parts by weight: 10 to 300 parts of the enteric polymer, surfactant The amount of the isotonicity adjusting agent is adjusted to adjust the pH of the enteric microcapsule preparation to 6.6 to 8.7, and the balance is water.
6. A method for preparing an enteric oxygen microcapsule for regulating intestinal flora according to any one of claims 1, 2, 3 or 4, comprising the steps of:

   1) preparing an aqueous solution of an emulsifier, a thickener, and a stabilizer, respectively;

   2) preparing an enteric cyst wall aqueous solution;

   3) The oxygen/oxygen source in the core material is passed through the aqueous solution of the emulsifier prepared in the step 1). After 1 hour, the chitosan and polyvinyl alcohol aqueous solution in the dissolved enteric wall are slowly added and stirred for 15 minutes. , adding yeast aqueous solution of powder, continue to stir and emulsify, and then slowly add aqueous sodium alginate solution in the enteric wall, ice bath and stir;

   4) adding a cross-linking agent dilution solution while stirring in an ice bath, and after the Weissenberg effect occurs, adjust the pH of the solution to disperse the solid into microcapsules, and then continue to stir for more than 10 minutes;

   5) The solution prepared above was sieved to remove large particles of impurities and precipitates; the supernatant was centrifuged, washed with a 5% sodium borohydride solution, and dried by suction filtration to obtain a product.
7. A method for preparing an enteric oxygen microcapsule for regulating intestinal flora according to any one of claims 1, 2, 3 or 5, comprising the steps of:

   1) soaking the enteric polymer to prepare an enteric coating solution;

   2) dissolving the isotonic adjusting agent and the surfactant in the auxiliary material with water or adding a co-solvent to uniformly disperse;

   3) combine the enteric coating liquid in step 1) and the auxiliary agent in step 2), add water or enteric polymer cosolvent to the full amount, stir at high speed, adjust the pH to 6.6 ~ 8.7;

   4) Pass the oxygen/oxygen source into the high-speed stirring solution of step 3) to form fine microcapsules containing a large amount of oxygen, and homogenize into stable oxygen microcapsules.
8. Use of an enteric oxygen microcapsule for regulating intestinal flora according to any one of claims 1 to 5, characterized in that the preparation of intestinal oxygen microcapsules as the sole for regulating intestinal flora is prepared Application in medicines of one of the active ingredients or active ingredients.
9. Use of an enteric oxygen microcapsule for regulating intestinal flora according to claim 8, wherein said enteric oxygen microcapsule is used for preparing a prophylactic or/and therapeutic and intestinal flora The use of drugs for imbalance-related diseases or drugs for mites.
10. Use of an enteric oxygen microcapsule for regulating intestinal flora according to claim 9, wherein said diseases associated with intestinal flora imbalance are obesity, diabetes, hypertension, cancer Or one or more of Alzheimer's disease.