WO2021115156A1

WO2021115156A1 - Composition containing sodium pyruvate and use thereof

Info

Publication number: WO2021115156A1
Application number: PCT/CN2020/133041
Authority: WO
Inventors: 周方强
Original assignee: 周方强
Priority date: 2019-12-10
Filing date: 2020-12-01
Publication date: 2021-06-17
Also published as: CN110859307A

Abstract

Disclosed are a composition containing sodium pyruvate for oral administration and the use thereof, wherein the composition comprises the following essential components: (a) 2.0-25.0 parts by weight of sodium pyruvate; and (b) 5.0-100.0 parts by weight of anhydrous glucose or other carbohydrates. The above-mentioned composition can be used to treat diabetes and organ complications thereof, or can be used to treat conditions involving increased pyruvate dehydrogenase (PDH) activity.

Description

Composition containing sodium pyruvate and its use

Technical field

The present invention relates to the field of drug therapy, in particular to an oral composition containing sodium pyruvate to treat diabetes and its organ complications and non-diabetic-induced pyruvate dehydrogenase (PDH) activity reduction related diseases.

Background technique

Diabetes is a common disease at home and abroad. According to statistics from the International Diabetes Federation, there were approximately 415 million patients worldwide in 2015; the domestic incidence in adults was approximately 10.9% in 2013 [1,2]; currently, there are more than 114 million adult diabetic patients in China. The rate is still rising. Although there are a variety of specific therapeutic drugs, there is still a lack of ideal drugs that act on multiple links in the pathogenesis; effective prevention and treatment measures are still lacking in a large area of population. Diabetes is a disorder of systemic glucose oxidation and metabolism, which leads to high blood sugar and long-term stimulation of high glucose in tissue cells, resulting in microvascular disease of the whole body. Although the damage of various organs has different manifestations, the basic pathophysiological changes are similar: accompanied by tissue hypoxia , Acidosis, oxidative stress and inflammation, as well as mitochondrial damage and apoptosis, as well as the production and tissue deposition of characteristic glycosylation end products (AGEs), which further promote the inflammatory response, leading to a vicious circle. The pathological changes develop irreversibly to organ dysfunction or failure; among them, diabetic kidney disease accounts for about 30-40% [3].

Traditionally, Pyruvate/propyl salt is not a stimulator of insulin secretion, but its derivative Methyl Pyruvate is. Since the end of the last century, there have been reports about the effects of pyruvate in rodent drinking water alleviating the production of end-products of glycation of the eye lens caused by high sugar in the body, alleviating cataracts caused by diabetes, and alleviating and protecting diabetic retinopathy. ; There are also in vitro experiments showing that pyruvate has the function of protecting the transplanted pancreatic tissue [4,5], but until 2007, Europe reported for the first time that human oral high-dose sodium pyruvate (about 1.0g/kg, orally after meals) ) Effectively alleviate the condition of 6 patients with type I (insulin-dependent) diabetes: even hypoglycemia occurs and the amount of insulin needs to be reduced [6]; there have been reports afterwards that oral pyruvate (0.5g/kg x three times a day) for half a year , Has the effect of directly stimulating the secretion of insulin by pancreatic islet cells, reducing the amount of insulin by about 1/3.

However, large doses of pyruvate (>10-15g) taken by humans at one time have obvious gastrointestinal irritation symptoms: abdominal pain, diarrhea, etc., which cannot be tolerated for a long time, so they cannot be used in clinical practice. At the same time, the literature confirms that human oral administration of a smaller dose of pyruvate, such as 7g/d x week, does not have any effective physiological effects [7]; a larger dose, such as a single oral administration of pyruvate 25g, only temporarily relieves the effects of exercise. Acidosis does not improve exercise endurance [8].

Therefore, there is an urgent need in the art for a kind of oral administration (gastrointestinal tract) that can exert pyruvate to relieve diabetes and similar disorders of glucose metabolism (their common point is the decrease of PDH activity), and at the same time, it can make the human gastrointestinal Road tolerance, suitable for clinical promotion of technical solutions for the function of pyruvate.

Summary of the invention

The present invention aims to provide a composition containing sodium pyruvate which can be used orally to prevent and treat diabetes or its organ complications and related diseases caused by or accompanied by decreased pyruvate dehydrogenase (PDH) activity.

In the first aspect of the present invention, a composition is provided, which includes the following essential components:

(a) 2.0-25.0 parts by weight of sodium pyruvate; and

(b) 5.0-100.0 parts by weight of anhydrous glucose or other carbohydrates;

And the weight of essential components accounts for 30-100% of the weight of the composition.

In another preferred embodiment, the composition has additional components, and the additional components include: sodium chloride, potassium chloride or potassium citrate, vitamin B, vitamin C, vitamin E and zinc preparations.

In another preferred embodiment, the composition contains the following components:

(i) 2.0-25.0 parts by weight of sodium pyruvate;

(ii) 0-10.0 parts by weight of sodium chloride;

(iii) 0-2.0 parts by weight of potassium chloride or the corresponding equivalent of potassium citrate; and

(iv) 5.0-100.0 parts by weight of anhydrous glucose or other carbohydrates;

And the weight of components (i)+(ii)+(iii)+(iv) accounts for 80-100% of the total weight of the composition.

In another preferred example, the molar ratio of sodium ions to glucose in the composition is 0.5-1:1; more preferably close to 1:1.

In the second aspect of the present invention, there is provided a composition provided by the present invention as described above in the preparation of a medicine, food or medicine for the treatment or prevention of diseases or disorders related to or accompanied by the inhibition of pyruvate dehydrogenase activity. Use in health products and functional drinks.

In another preferred example, the disease is diabetes and its organ complications.

In another preferred embodiment, the diabetes is type II diabetes, and/or type I diabetes.

In another preferred embodiment, the drug, food or health product and functional beverage are in the form of a solution; preferably, the osmotic pressure of the solution is 200-1200 mOsm/L, and more preferably, the osmotic pressure of the solution is 220-280mOsm/L, or 600-1000mOsm/L.

In another preferred example, the disease or condition also includes some critical illnesses, infections, tumors and senile diseases, etc. or accompanied by the performance of glucose metabolism disorders caused by decreased pyruvate dehydrogenase activity in the body; for example, but not limited to , Shock of various etiologies, bacterial or viral inflammation, burns or colon and pancreatic tumors, and Alzheimer’s.

Accordingly, the present invention provides a function of oral (gastrointestinal) or intravenous injection that can exert pyruvate to relieve diabetes and similar disorders of glucose metabolism (their common point is the decrease of PDH activity), and at the same time, it can It can make the human gastrointestinal tract tolerate, and is suitable for clinical promotion of the technical plan for the function of pyruvate.

Description of the drawings

Figure 1 shows the effects of oral administration of two sodium pyruvate aqueous solutions on the activities of various enzymes in the kidney tissue of diabetic mice; among them,

A represents the effect on AR, or aldose reductase;

B represents the effect on PDH, which is pyruvate dehydrogenase;

C represents the effect on PDHA1, the non-phosphorylated active part of pyruvate dehydrogenase;

D represents the effect on PDK, which is pyruvate dehydrogenase kinase;

Nor represents the control group of normal mice;

Con represents the untreated control group of diabetic mice;

Pyr refers to the treatment group of diabetic mice with an aqueous solution containing only 1w/v% sodium pyruvate orally;

Pyr-ORS means oral sodium pyruvate composition (0.35w/v%NaPyr, 0.2w/v%NaCl, 0.15w/v%KCl, 1.35w/v%Glucose diabetic mice treatment group;

** indicates that there are significant statistical differences between the two treatment groups and the control group of untreated mice.

The chart shows that oral administration of two sodium pyruvate solutions in diabetic mice also effectively inhibits the increased AR and PDK activities in the body (kidney tissue) stimulated by high glucose, and promotes the complete recovery of the PDH activity inhibited by PDK.

Detailed ways

The inventor’s research found that a composition containing sodium pyruvate and glucose can treat diabetes and its organ complications in a human diabetes (type II) animal model (db/db mice) by oral administration, and the treatment of sodium pyruvate is effective The amount is the amount that is sufficiently tolerated by the human body, so that it can be promoted and used in clinics and/or the population. Furthermore, the inventors also confirmed for the first time that such a composition can significantly increase the PDH activity of one of the key enzymes of inhibited sugar oxidation metabolism in diabetic animals by oral administration (this is the first time in the world to confirm in this experiment), and at the same time in animals It is the first proof that the oral sodium pyruvate composition also restores the activity of pyruvate kinase (PK), one of the key glycolysis enzymes inhibited by hyperglycemia, improves the oxidation/reduction potential in organs and tissues, and significantly increases insulin in animal plasma It was also found in in vivo and in vitro experiments that pyruvate can inhibit the aldose reductase (AR) activity that is abnormally increased by high glucose stimulation, which may reverse the abnormally enhanced glycolytic sorbitol branch of hyperglycemia [9 ], effectively restore the close to normal glycolysis process and oxidative metabolism of sugar in diabetic animals (that is, reverse the Warburg phenomenon of abnormal sugar metabolism in diabetes: under aerobic conditions, the end products of glycolysis increase, while the oxidative metabolism of sugar is inhibited [10] ), while reducing fasting high blood sugar levels, reducing body weight, and improving the diabetic state of animals; inhibiting/reducing the oxidative stress response and the production of AGEs in the kidney tissue. Oxidative stress/inflammatory response in the organs and tissues of diabetes and the generation/accumulation of glycation end products are the two key factors for the occurrence and development of diabetes/organ complications besides high glucose, leading to a vicious cycle of disease [3]. Therefore, oral administration of the composition provided by the present invention improves the function and pathological changes of organ complications (kidney). The experiment of the present invention demonstrates that the oral composition containing a small dose of sodium pyruvate can effectively reverse the decreased PDH activity and the Warburg phenomenon, and the experimental results are consistent with the clinical curative effect, and the present invention is completed on this basis. The present invention solves the problem of using a low-dose and tolerable sodium pyruvate composition to achieve the clinically unacceptable high-dose sodium pyruvate to prevent and treat diabetes and organ complications, and to promote the prevention and treatment of diabetes and other PDHs associated with the above-mentioned PDH in large-scale populations. The purpose of suppressing related disorders.

combination

The present invention provides a composition in which sodium pyruvate and anhydrous glucose or other carbohydrates are essential components. The composition may also contain sodium chloride, potassium chloride or potassium citrate, vitamin B, and vitamin C. , Vitamin E and zinc preparations and other additional components.

The content of essential components in the composition provided by the present invention can be (a) 2.0-25.0 parts by weight of sodium pyruvate; and (b) 5.0-100.0 parts by weight of anhydrous glucose or other carbohydrates;

The amount of sodium pyruvate can be 2.0-6.0 parts by weight, 2.0-10.0 parts by weight, 3.5-15.0 parts by weight, 5.0-20.0 parts by weight, etc.;

The amount of anhydrous glucose or other carbohydrates can be 7.0-50.0 parts by weight, 20.0-90.0 parts by weight, 40.0-75.0 parts by weight, 12.0-20.0 parts by weight, 30.0-80.0 parts by weight, etc.

According to the description of the present invention, those skilled in the art can understand that, as long as the composition provided by the present invention can achieve sufficient solubility, for example, can absorb a sufficient and effective amount of pyruvate, then the essential component (a) component of pyruvate can be added Sodium is replaced by pyruvate formed by pyruvate and other inorganic bases, such as but not limited to calcium pyruvate. In some embodiments of the present invention, based on the total weight of component (a), about 0%-100% calcium pyruvate is used instead of sodium pyruvate, for example, but not limited to, 5%-90%, 45%-85% %, 15%-80%, 40%-75%, 35%-70%, 30%-65%, 25%-95%, 50%-98%, 20%-60%, 10%-55%, etc. Calcium pyruvate replaces sodium pyruvate.

In some embodiments, the composition provided by the present invention contains: (i) 2.0-25.0 parts by weight of sodium pyruvate; (ii) 0-10.0 parts by weight of sodium chloride; (iii) 0-2.0 parts by weight of potassium chloride Or the corresponding equivalent of potassium citrate; and (iv) 5.0-100.0 parts by weight of anhydrous glucose or other carbohydrates; the composition may also contain sodium citrate, but the composition does not contain sodium bicarbonate in principle.

The amount of sodium pyruvate can be 2.0-6.0 parts by weight, 3.0-4.0 parts by weight, 2.0-10.0 parts by weight, 3.5-15.0 parts by weight, 5.0-20.0 parts by weight, etc.;

The amount of sodium chloride can be 0-5.0 parts by weight, 2.0-3.5 parts by weight, 4.0-8.0 parts by weight, etc.;

The amount of potassium chloride or potassium citrate can be 0-1.8 parts by weight, 1.5-2.0 parts by weight, etc.;

The amount of anhydrous glucose or other carbohydrates can be 7.0-50.0 parts by weight, 20.0-90.0 parts by weight, 13.5-50.0 parts by weight, 40.0-75.0 parts by weight, 12.0-20.0 parts by weight, 30.0-80.0 parts by weight, etc.

In some embodiments, the molar ratio of sodium ion to glucose in the composition provided by the present invention is 0.5-1:1, such as 0.6-0.8:1 and 0.7-0.9:1. In a preferred embodiment, the molar ratio of sodium ion to glucose in the composition provided by the present invention is close to (approximately) 1:1, such as 0.6-0.99:1; usually, it does not exceed 1:1.

The composition provided by the present invention may be in a solid form, such as but not limited to a powder form; it may also be in a liquid form, such as, but not limited to, a drinking water solution. After sterilization, it can be prepared into aqueous solution injection for intravenous injection.

If the composition provided by the present invention is to be stored in liquid form at room temperature, the stability of the aqueous solution of sodium pyruvate needs to be considered, and the pH of the solution is 3.5-4.9.

As used in the present invention, "other carbohydrates" include hydrous glucose, fructose, sucrose, starch, and cereals. The cereals include rice, millet, corn, sorghum, wheat and the like. The cereals used in the present invention may be powdered food products, such as, but not limited to, rice flour, corn flour, sorghum flour and the like. In actual applications, the amount of rice noodles that replace or partially replace 20 grams of anhydrous glucose is usually 50-80 grams.

use

The composition provided by the present invention can be used to treat or prevent diabetes and its organ complications. Generally, it is necessary to prepare medicines, foods or health products and functional beverages from the provided composition, and then give the individual an effective amount of the medicine when needed. The composition provided by the invention.

The composition provided by the present invention can also be used for oral administration (including gastrointestinal tract) or intravenous injection to increase the activity of PDH for other related diseases except diabetes. Similarly, it is generally necessary to prepare medicines, foods or health products and functional beverages from the provided compositions, and to give individuals a certain effective amount of the compositions provided by the present invention when needed.

When the composition provided by the present invention is used to treat or prevent diabetes and its organ complications, or to improve the activity of PDH, the most suitable and effective amount for adults, based on the sodium pyruvate contained therein, is 5-10 (not More than 15) g/day.

In some embodiments, the diabetes is type II diabetes, and/or type I diabetes; the diabetic organ complications include, but are not limited to, diabetic nephropathy, diabetic eye complications, diabetic foot, and diabetic cardiovascular complications , Diabetic cerebrovascular disease, diabetic neuropathy, etc.

Inhibition of PDH activity in tissue cells is the acute and chronic organ damage caused by many pathogenic factors, including diabetes and diabetic ketoacidosis, and its organ damage [11,12]; a variety of critical diseases caused by non-diabetics, such as hemorrhage , Trauma or infection and the shock caused by this, even tumors, congenital hypoplasia of the brain and senile degeneration, etc., all have abnormal glucose metabolism caused by low PDH activity: that is, the metabolic changes characterized by the Warburg phenomenon, which are them One of the common characteristics of cell metabolism and one of the important factors of the molecular mechanism of cell metabolism [10,13,14]. The formation and accumulation of AGEs is not only a complication of diabetic organs, but also an important pathogenic factor of senile neurodegenerative degeneration (Alzheimer’s Dis). The composition provided by the present invention improves the progress of various diseases by increasing the activity of PDH. For example, diabetic ketoacidosis is a critical illness, and its PDH activity in the body is lower than that of non-ketoacidosis diabetic patients [12]; Since pyruvate itself is a powerful antacid preparation, it can effectively correct fatal hypoxic lactic acidosis either by intravenous injection or oral administration [15], so it is also suitable for the above-mentioned diabetic ketoacidosis and non-diabetic Other related diseases. Therefore, if the conditions for intravenous administration are not available, it is more suitable for oral administration of medicines, foods or health products and functional drinks obtained from the composition provided by the present invention.

The inventors used in 1000 ml of water containing 2.0-6.0 grams of sodium pyruvate, 1.5-2.0 grams of sodium chloride, 0-1.5 grams of potassium chloride or the corresponding equivalent of potassium citrate and 5.0-50.0 grams of anhydrous glucose or other corresponding The amount of carbohydrate aqueous solution, in which, 0.35w/v% sodium pyruvate composition aqueous solution (0.35% NaPyr, 0.2% NaCl, 0.15% KCl, 1.35% Glucose, osmotic pressure about 247mOsm/) was used to feed diabetic mice. Model, and used an aqueous solution containing 1w/v% simple sodium pyruvate as a positive control, and was used in an experimental model of diabetic mice at the same time. As a result, it was found that the aqueous solution formed by the composition provided by the present invention can be compared with the positive control (1% sodium pyruvate) The aqueous solution is also effective in pre-curing the progress of diabetes in mice, and is also effective in improving the activity of pyruvate dehydrogenase. Compared with the control group of non-treated diabetic mice, these effects have statistically significant differences.

Taking into account the human body’s reasonable daily water intake, in some embodiments, an aqueous solution containing 20-25 grams of sodium pyruvate and 60.0-100.0 grams of anhydrous glucose or other corresponding amounts of carbohydrates in 1000 ml of water may be used, which may not contain Sodium chloride and potassium chloride, or 0-2.0 grams of sodium chloride and 0-2.0 grams of potassium chloride or the corresponding equivalent of potassium citrate, the osmotic pressure is less than 1000mOsm/L. It can also effectively treat and prevent diabetes and its organ complications, and can also effectively improve the activity of pyruvate dehydrogenase, and it can be tolerated by the human body / gastrointestinal tract without side effects. When the human body takes 250-500ml of the above-mentioned hypertonic formula solution orally every day, it is equivalent to oral 5-10g/d sodium pyruvate. The intake of such a volume of solution is only approximately equivalent to the volume of a bottle of beverage can. When the composition provided by the present invention is mixed with a food acceptable carrier, the resulting functional beverage can be administered to the population, thereby improving Widely and effectively play a role in the treatment and prevention of diabetes and related diseases.

In some embodiments, the composition provided by the present invention can also be contacted with the PDH inhibited by high sugar or other pathogenic factors, thereby reversing/restoring the inhibited PDH activity. Obviously, this method can be carried out in in vitro experimental studies for different purposes, which has also been confirmed in the in vitro cell experiments in the high-sugar environment of the present invention.

Another use of the composition provided by the present invention is for in vitro screening experiments, for screening and verifying ingredients for the treatment or prevention of diabetes and its organ complications; or for screening and verifying ingredients for improving the activity of PDH.

The composition provided by the present invention is used in a screening experiment, specifically the composition provided by the present invention is used in a screening experiment to verify candidate compounds. These candidate compounds can be used to treat or prevent one or more diabetes and its organ complications . Alternatively, the composition provided by the present invention can be used to confirm the ability of a candidate compound to increase PDH activity.

The method for preparing medicines (preparations), food or health products and functional beverages from the composition provided by the present invention includes the steps of: taking the composition provided by the present invention as an active ingredient and forming a product with a pharmaceutically or food acceptable carrier.

Taking the preparation of a drug (preparation) from the composition provided by the present invention as an example, suitable administration routes of the drug (or sterile preparation) include, but are not limited to, oral administration, transgastric tube or surgical gastrointestinal and rectal administration. medicine. For example, but not limited to, formulating oral drugs, mixing 5-10g sodium pyruvate with 20-40g glucose or corresponding starch, the mixture can be made into oral powder, or other solid preparations such as tablets or capsules suitable for oral administration ; Or by intravenous injection.

When the pharmaceutical preparation is an aqueous solution, the osmotic pressure of the solution is 200-1200 mOsm/L, for example, 200-280 mOsm/L, 240-800 mOsm/L, 400-900 mOsm/L, or 600-1000 mOsm/L. The aqueous solution is stored in liquid form at room temperature, and the stability of the aqueous solution of sodium pyruvate needs to be considered. The pH of the general solution is 3.5-5.0.

When treating or preventing diabetes or its organ complications or increasing the level of inhibited PDH in the body, the dosage of the composition provided by the present invention is calculated as sodium pyruvate, usually 5-10g/day (it can also be 7-9g/day, 6-8g/day, etc.), the administration period is 1-8 weeks or longer. In some embodiments, it includes multiple administrations of a certain effective amount of the composition provided by the present invention. These examples specifically include: i) single-dose administration of the composition provided by the present invention; ii) multiple administrations with a time interval of 6-8 hours. In a further or alternative preferred embodiment, the method has a drug holiday, specifically referring to the temporary delay of the administration of the composition provided by the present invention or the temporary reduction of the dose of the composition provided by the present invention; during the drug holiday At the end, resume the dosage of the composition provided by the present invention. The duration of the drug holiday varies from two days to one year.

the term

The term "an effective amount" means that for the purpose of treating diabetes, this amount can achieve the desired effect of lowering blood sugar and/or reducing the amount of insulin used after a proper administration period.

The term "acceptable" refers to a prescription component or active ingredient that does not have undue harmful effects on the health of the general treatment or prevention target.

As used herein, the term "prevention" refers to the ability to delay, or stop, or even partially reverse the progression of diabetes-prone (pre-stage) or diabetics and their organ complications and related disorders with decreased PDH activity.

As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier used for the administration of a therapeutic agent, and includes various excipients and diluents. The term refers to such pharmaceutical carriers: they are not essential active ingredients themselves, and they are not excessively toxic after administration. Suitable carriers are well known to those of ordinary skill in the art. A full discussion of pharmaceutically acceptable excipients can be found in Remington's Pharmaceutical Sciences (Mack Pub. Co., N.J. 1991). Pharmaceutically acceptable carriers in the composition may include liquids such as water and saline. In addition, these carriers may also contain auxiliary substances, such as disintegrants, emulsifiers, and pH buffer substances. Other additional components are also included in the definition of pharmaceutically acceptable carriers. For example, the composition of the present invention may also include, but is not limited to, vitamin B, vitamin C, vitamin E, zinc preparations, organic acids: such as acetate. , Malate, amino acids: such as arginine, alanine and taurine, edible coloring, flavoring, etc.

The terms "subject" or "patient" are used interchangeably herein, which refers to animals (including humans) that can be treated by the compound and/or method. "Individual" or "patient" herein encompasses both males and females, unless specifically stated otherwise. Therefore, "individual" or "patient" includes any mammals, including, but not limited to, humans, non-human primates, such as mammals, dogs, cats, horses, sheep, pigs, cows, etc., which can be used for The compounds benefit from treatment. Animals suitable for treatment with the compounds and/or methods of the present invention are preferably humans. Generally speaking, the term "patient" and the term "individual" can be used interchangeably in this article.

Although the numerical ranges and parameters used to define the broader scope of the present invention are approximate numerical values, the relevant numerical values in the specific embodiments are presented here as accurately as possible. However, any value inevitably contains standard deviations due to individual test methods. Here, "about" usually means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a specific value or range. Alternatively, the term "about" means that the actual value falls within the acceptable standard error of the average, depending on the consideration of those skilled in the art. In addition to the experimental examples, or unless otherwise clearly stated, all ranges, quantities, values and percentages used herein (for example, used to describe the amount of material, length of time, temperature, operating conditions, quantity ratios and other similar Those) have been modified by "about". Therefore, unless otherwise stated to the contrary, the numerical parameters disclosed in this specification and the appended claims are approximate values, and can be changed as required. At least these numerical parameters should be understood as the indicated effective number of digits and the value obtained by applying the general carry method.

Unless otherwise defined in this specification, the scientific and technical terms used herein have the same meanings as understood and used by those skilled in the art. In addition, without conflict with the context, the singular nouns used in this specification cover the plural nouns; and the plural nouns used also cover the singular nouns.

The above-mentioned features mentioned in the present invention or the features mentioned in the embodiments can be combined arbitrarily. All the features disclosed in the specification of this case can be used in combination with any composition form, and each feature disclosed in the specification can be replaced by any alternative feature that can provide the same, equal or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equal or similar features.

The main advantages of the present invention are:

1. For the first time, in addition to existing diabetes treatment drugs, sodium pyruvate can be used orally to promote the use of sodium pyruvate in the treatment of diabetes and its organ complications;

2. Since pyruvate itself is a powerful antacid preparation and a stimulant of PDH, it is very suitable for the prevention and treatment of diabetic ketoacidosis, and it may even be taken by mouth without the application of insulin;

3. In patients with clinical diabetic tendency, promotion and application without special dietary restrictions can achieve the effect of preventing and treating diabetes and organ complications;

4. For the first time, it provides the possibility and simple specific methods that can be promoted and applied in large-scale population to prevent and treat diabetes and organ complications;

5. In addition to being developed as an innovative drug, it can also be promoted in the population as a physiological liquid supplement or a health-care performance "functional drink", to give full play to the possibility and specificity of preventing diabetes and organ complications in large-scale populations. method.

6. It is also only suitable for oral PDH inhibition caused by other causes of non-diabetics, such as multi-cause shock, chronic organ dysfunction, infection (including infectious diseases), and even tumors and senile diseases under certain conditions , Can also play a certain degree of prevention and treatment in the population.

The present invention will be further explained below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods that do not indicate specific conditions in the following examples are usually in accordance with conventional conditions or in accordance with the conditions recommended by the manufacturer. Unless otherwise stated, all percentages, ratios, ratios, or parts are by weight.

The unit of weight-volume percentage in the present invention is well known to those skilled in the art, for example, refers to the weight of the solute in a 100 ml solution.

Unless otherwise defined, all professional and scientific terms used in the text have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to the content described can be applied to the method of the present invention. The preferred implementation methods and materials described in this article are for demonstration purposes only.

Example

Preparation of reagents:

Drug group: 0.35% sodium pyruvate, 0.2% sodium chloride, 0.15% potassium chloride and 1.35% glucose formula aqueous solution, instead of conventional animal laboratory drinking water;

Normal group: routine laboratory animal drinking water;

Control group: 1.0% sodium pyruvate aqueous solution, instead of drinking water in the conventional animal laboratory.

Experimental Example 1

Experimental animals:

Normal group: C57BLKS/J normal untreated mice: drinking regular laboratory drinking water;

Control group: C57BLKS/J diabetic db/db untreated mice (conventional human type II diabetes animal experimental model): drinking regular laboratory drinking water;

1% Pyr: Drink db/db mice containing pure 1% sodium pyruvate distilled water to replace regular laboratory drinking water;

Drug group: Drinking an aqueous solution containing the sodium pyruvate composition of the present invention to replace conventional laboratory drinking water for db/db mice, which contains 0.35% sodium pyruvate, 0.2% sodium chloride, and 0.15% chlorination Potassium and 1.35% glucose.

experimental method:

The above randomly selected diabetic mice (10 weeks old) were divided into three groups, and the normal mice of the same strain drank different experimental water, and observed and recorded under the same experimental conditions: body weight, drinking water amount (about 10ml per day on average, No difference between groups) and urine output. The experimental observation was from 8 weeks to 18 weeks of age. After execution, the blood was taken for laboratory tests and the kidneys were taken for further examination. The following are the main experimental results and explanations observed by routine or commonly used laboratory testing methods today.

statistical methods:

^* P<0.05 and ^** P<0.01 are compared with the normal group;

^# P<0.05 and ^## P<0.01 are compared with the control group;

The values of diabetic db/db mice at baseline (10 weeks of age) and 8 weeks (18 weeks of age) are mean±standard deviation, n=6 in each group.

Result data:

1. The effect of different treatment methods on body weight, blood sugar and plasma insulin levels in diabetic mice

Table 1

Note: Glucose refers to fasting blood glucose; insulin refers to fasting plasma insulin.

The results showed that the oral administration of two different sodium pyruvate drinking water can also effectively control the weight gain of experimental animals, maintain fasting blood glucose levels no longer increase, and the plasma insulin level doubled; it clearly shows that both sodium pyruvate solutions are both So that the diabetes state can be effectively controlled.

2. Changes in serum cystatin C, 24-hour urine protein and NGAL after different treatments

Table 2

Note: NAGL, neutrophil gelatinase-related lipocalcin in urine.

The results showed that oral administration of two different sodium pyruvate drinking water can also significantly reduce the amount of 24-hour urine protein; reduce the level of cystatin C in the serum and the content of NAGL in the urine. The latter two determinations are early sensitive indicators of renal damage. It clearly shows that both sodium pyruvate solutions are equally effective in protecting/reducing diabetic kidney damage.

3. Changes of ROS, SOD, CAT, MDA and GSH-PX in kidney homogenate after different treatments

table 3

Note: ROS means reactive oxygen species; SOD means superoxide dismutase; CAT means catalase; MDA means malondialdehyde; GSH-PX means glutathione peroxidase.

The results show that the above multiple indicators reflecting the oxidative stress response in the body all show: oral administration of two different sodium pyruvate drinking water can also significantly inhibit the indicators of pro-oxidative stress response ROS and MAD; increase the antioxidant SOD, CAT and Antioxidant enzyme activity such as GSH-PX. Oxidative stress response and the inflammatory response promoted thereby are an important internal link in the development of diabetes. Oral sodium pyruvate effectively inhibits the oxidative stress response in diabetic mice and helps control diabetes and nephropathy.

4. Changes of LDH, PK and PDHa/PDHt in kidney homogenate after different treatments

Table 4

Note: LDH is lactate dehydrogenase; PK is pyruvate kinase; PDHa/PDHt is the active part of pyruvate dehydrogenase/total amount of pyruvate dehydrogenase.

The results showed that the activities of PK, one of the key enzymes of glycolysis and the key enzyme of sugar oxidative metabolism, PDH, which were inhibited in diabetic mice, were significantly increased after oral administration of sodium pyruvate. Therefore, it promoted the process of hyperglycemia into normal oxidative metabolism. And effectively control blood sugar and the metabolism and function of the body's organs; at the same time, AR and LDH stimulated by high glucose are inhibited, which promotes the abnormal glucose metabolism characteristics in diabetes: the reversal of the Warburg phenomenon (Warburg Effect) [10].

^{5. Changes of AGEs and NAD +} /NADH in kidney homogenate after different treatments

table 5

Note: AGEs are advanced glycosylation end products; NAD ⁺ /NADH is nicotinamide adenine dinucleotide (oxidized form/reduced form).

The results show that oral administration of two sodium pyruvate aqueous solutions in diabetic mice can equally effectively increase the important redox potential energy in the body (kidney): NAD ⁺ /NADH ratio, which is beneficial to the body to restore anti-oxidative stress and inhibit inflammation, and significantly reduce The formation and accumulation of end-products of glycosylation in the body/tissues, the latter being the key factor in the progression and deterioration of the course of complications of the microvessels and organs in the body of diabetes, causing the vicious circle.

The results of the determination of multiple enzyme activities in kidney tissue by the Western Blot method in routine experiments are shown in Figure 1. The results in Figure 1 show that oral administration of two sodium pyruvate aqueous solutions in diabetic mice is equally effective in improving the recovery of PDH and PDHA1 activities in the body (kidney tissue). This is due to the inhibition of PDK that is abnormally hyperactive under high glucose; the two types contain Sodium pyruvate solution can also significantly reduce the AR activity enhanced by high sugar.

In this regard, the above multiple experimental results confirmed that: oral glucose-containing only 0.35% sodium pyruvate composition and 1.0% sodium pyruvate aqueous solution have the same curative effect on diabetes and renal complications in diabetic mice; also in hyperglycemic conditions Comprehensively improve the abnormality of multiple pathways of sugar metabolism and return to normal, and promote the secretion of insulin and improve the production of AGEs.

discuss:

Rodents oral administration of simple sodium pyruvate, in fact, is always accompanied by food, which is equivalent to oral sodium pyruvate containing glucose, and is essentially equivalent to oral sugar-containing sodium pyruvate composition. Therefore, it has shown the same curative effect in experiments. However, there is no need for 1.0% and only 0.35% sodium pyruvate is sufficient to achieve the same effect; however, in humans, oral sodium pyruvate (7-25 g/day) is usually ineffective, and the blood concentration does not increase due to intestinal absorption. Poor, unless a large dose is used, but gastrointestinal irritation cannot be tolerated and promoted. Since the intestinal epithelial cells possess a sodium-glucose co-transporter (Na ⁺ -Glucose Co-transporter) [15], oral administration of a small amount of sodium pyruvate is due to the composition of the present invention containing glucose at the same time. The same effect can be achieved by taking a large oral dose. This is the gist of the present invention.

Experimental Example 2

Cell experiment

Human renal tubular epithelial cell line: HK-2 cells in high glucose (30mmol/L) and normal levels (5mmol/L) with 0.5mmol/L sodium pyruvate (5 times the concentration of normal serum levels: previous experiments It has been confirmed that oral administration of the above sodium pyruvate composition can increase the level of pyruvate by more than 5 times in animal plasma[15]) and incubate together for 3 days. The determination method of routine experiment: Western blot method to determine ARPDH/PDHA1( The main active components in PDH) and the activity of pyruvate dehydrogenase kinase (PDK), and the results showed that they were consistent with the above in vivo experimental results: inhibiting the enhanced AR and PDK, and restoring the activities of PDH and PDHA1.

It is worth noting that PDK in human cells will be stimulated and enhanced by many other pathogenic factors besides high glucose, such as hypoxia, oxidative stress, tumors or aging; PDK is the kinase of PDH, which can spontaneously undergo phosphoric acid. It inhibits the activity of PDH (PDH is phosphorylated and loses its activity, and the ratio of PDHa/PDHt decreases, as shown in the above results), and there is a disorder of glucose metabolism: a performance characterized by the Warburg phenomenon. Both in vivo and in vitro experiments of the animals of the present invention have confirmed that sodium pyruvate exerts an inhibitory effect on AR and PDK activities enhanced by abnormal stimulation, and restores the activity of PDH. The above results are in line with the consensus that pyruvate activation of PDH is the result of inhibiting PDK (PDH is directly regulated by the phosphorylation inactivation of PDK) [16], confirming that oral administration of the sodium pyruvate composition provided by the present invention can effectively enhance inhibition in vivo PDH activity.

discuss:

Pyruvate is a ‘PDH stimulant’ typically represented by dichloroacetic acid (DCA) [16-18]. The above description: oral administration of the sodium pyruvate composition of the present invention restores the inhibited PDH activity in the renal tissue, because the absorbed pyruvate directly inhibits the abnormally increased PDK with the accompanying sugar [16]. Although pyruvate can increase the inhibited PDH activity whether administered systemically in vivo or in vitro, the experiments of the present invention have demonstrated for the first time that only a limited dose of sodium pyruvate composition can effectively stimulate the recovery of enhanced PDK in vivo. Inhibit the activity of the key enzyme PDH, thereby improving abnormal glucose metabolism in the whole body. Therefore, oral administration is also applicable to a variety of non-diabetic diseases with decreased PDH activity, such as shock, trauma, infection, tumor and elderly patients: they all show that the phosphorylation of PDK in the tissues leads to a decrease in PDH activity. Experiments have shown that intravenous/intraperitoneal administration of large doses of sodium pyruvate has a considerable therapeutic effect [19-22]. Therefore, the expected effect can also be achieved by using the sodium pyruvate composition of the present invention for oral administration.

All documents mentioned in the present invention are cited as references in this application, as if each document was individually cited as a reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

references:

1, Ogurtsova K, et al. IDF Diabetes Atlas: Global estimates for the prevalence of diabetes for 2015 and 2040. Diabetes Res Clin Pract. 2017Jun; 128: 40-50.

2, Wang L,et al. Prevalence and Ethnic Pattern of Diabetes and Prediabetes in China in 2013.JAMA.2017Jun 27; 317(24): 2515-2523.

3. Aliic RZ, et al. Diabetic Kidney Disease: Challenges, Progress, and Possibilities. Clin J Am Soc Nephrol. 2017Dec 7; 12(12): 2032-2045.

4. Devamanoharan PS, et al. Non-enzymatic glycation of lens proteins and haemoglobin-inhibition by pyruvate: an in-vivo study. Diabetes Obes Metab. 1999 May; 1(3): 159-64.

5. Brown M, et al. Pyruvate improve pancreatic islet engraftment and functionality. Transplant Proc. 2001 Nov-Dec; 33(7-8): 3523.

6, Petkova I, et al. Oral application of sodium pyruvate in healthy persons and patients with diabetes mellitus type 1. Comptes rendus de l'Acad_emie bulbare des Sciences. 2007; 60: 579-84.

7, Morrison MA, et al. Pyruvate ingestion for 7 days does not improve aerobic performance in well-trained individuals.J Appl Physiol(1985).2000Aug;89(2):549-56.

8,Olek RA,et al.Single pyruvate intake induces blood alkalization and modification of resting metabolism in humans.Nutrition.2015Mar;31(3):466-74.

9, Yan LJ. Redox imbalance stress in diabetes mellitus: Role of the polyol pathway. Animal Model Exp Med. 2018Mar; 1(1): 7-13.

10. Zhang G, et al. The Warburg effect in diabetes kidney disease. Semin Nephrol. 2018; 38(2): 111-120.

11. Mostert M, et al. Derangements of pyruvate dehydrogenase in circulating lymphocytes of NIDDM patients and their healthy offspring. J Endocrinol Invest. 1999; 22(7):519-26.

12. Andersen LW, et al. Pyruvate dehydrogenase activity is decreased in emergency department patients with diabetes ketoacidosis. Acad Emerge Med. 2016; 23(6):685-9.

13, Luo F, et al. Hexokinase II promotes the Warburg effect by phosphorylating alpha subunit of pyruvate dehydrogenase. Chin J Cancer Res. 2019Jun; 31(3):521-532.

14, Burns JS, et al. Metabolic pathways of the Warburg effect in health and disease: Perspectives of choice, chain or chance. Int J Mol Sci. 2017Dec 19; 18(12).pii: E2755.

15,Liu,R,et al.Pyruvate in reduced osmolarity oral rehydration salt corrected lactic acidosis in severe rats.J.Surg.Res.2018;226:173-180.

16,Priestman DA,et al.Pyruvateinhibition of pyruvate dehydrogenasekinase.FEBS Lett.1996Sep 16;393(2-3):174-8.

17, Whitehouse S, et al. Mechanism of activation of pyruvate dehydrogenase by dichloroacetate and other halogenated carboxylic acids. Biochem J. 1974 Sep; 141(3): 761-74.

18, Jaimes R 3rd, et al. Functional response of the isolated, perfused normoxic heart to pyruvate dehydrogenase activation by dichloroacetate and pyruvate.Pflugers Arch. 2016Jan; 468(1):131-142.

19, Sharma P, et al. Pyruvate modulates hepatic mitochondrial functions and reduces apoptosis indicators during hemorrhagic shock in rats.Anesthesiology.2005Jul;103(1):65-73.

20. Sharma P, et al. Role of pyruvate dehydrogenase complex in traditional brain injury and measurement of pyruvate dehydrogenase enzyme by dipstick test. J Emerge Trauma Shock. 2009 May; 2(2): 67-72.

21, Takakusagi Y, et al. Pyruvate induce transient tumor hypoxia by enhancing mitochondrial oxygen consumption and potentiates the anti-tumor effect of a hypoxia-activated prodrug TH-302.PLoS e One. 20149(107).

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Claims

A composition, characterized in that the composition includes the following essential components:

(a) 2.0-25.0 parts by weight of sodium pyruvate; and

(b) 5.0-100.0 parts by weight of anhydrous glucose or other carbohydrates;

And the weight of essential components accounts for 30-100% of the weight of the composition.
The composition of claim 1, wherein the composition has additional components, and the additional components include: sodium chloride, potassium chloride or potassium citrate, vitamin B, vitamin C, Vitamin E and zinc preparations.
3. The composition of claim 2, wherein the composition contains the following components:

(i) 2.0-25.0 parts by weight of sodium pyruvate;

(ii) 0-10.0 parts by weight of sodium chloride;

(iii) 0-2.0 parts by weight of potassium chloride or the corresponding equivalent of potassium citrate; and

(iv) 5.0-100.0 parts by weight of anhydrous glucose or other carbohydrates;

And the weight of components (i)+(ii)+(iii)+(iv) accounts for 80-100% of the total weight of the composition.
The composition according to any one of claims 1 to 3, wherein the molar ratio of sodium ion to glucose in the composition is 0.5-1:1; preferably close to 1:1.
A use of the composition according to any one of claims 1 to 4 in the preparation of medicines, foods or health products and functional beverages for treating or preventing diseases or disorders related to the inhibition of pyruvate dehydrogenase activity.
The use according to claim 5, wherein the disease is diabetes and its organ complications.
The use according to claim 5, wherein the diabetes is type II diabetes, and/or type I diabetes.
The use according to any one of claims 5-7, wherein the medicine, food or health product and functional beverage are in the form of a solution; the pH of the solution is 3.5-4.9.
The use according to claim 8, wherein the osmotic pressure of the solution is 200-1200 mOsm/L, preferably, the osmotic pressure of the solution is 220-280 mOsm/L, or 600-1000 mOsm/L.
The use according to claim 5, wherein the diseases or conditions also include some critical illnesses, infections, tumors and senile diseases, etc., which are caused by or accompanied by glucose metabolism disorders caused by decreased pyruvate dehydrogenase activity in the body. which performed.