WO2022089133A1 - Combination for lowering blood sugar, application, and method - Google Patents

Abstract

Provided is a combination for lowering blood sugar, comprising lysozyme and dosmalfate. Also provided are an application of the composition and a method for the composition. Lysozyme and dosmalfate have significant synergistic effects in lowering blood sugar, and can improve gastrointestinal dysfunction of hyperglycemic patients. In addition, the combination also has synergistic and attenuating effects on other hypoglycemic drugs, and therefore has a particular clinical value for blood sugar lowering and hyperglycemia related diseases or symptoms.

Description

A combination, application and method for lowering blood sugar technical field

The invention belongs to the field of medicine, and in particular relates to a combination, application and method for reducing blood sugar.

Background technique

Glucose is the main energy source for normal physiological activities of the human body. After the food is digested by the gastrointestinal tract, it is partially converted into glucose, absorbed into the blood, and supplied to various organs and tissues of the body through the blood circulation. Maintaining a certain blood sugar level is the key to the body's physiological functions. Under normal circumstances, the blood glucose concentration of the human body will remain at a relatively stable level, and the fasting blood glucose (FPG) of adults is about 4-7 mmol/L. Hyperglycemia is a common pathological state.

Fasting blood glucose above 6 mmol/L reflects glucose metabolism disorder, which is called prediabetes.

Fasting blood glucose greater than 7mmol/L or glycosylated hemoglobin (HbA1c) greater than 6.5% can be diagnosed as diabetes. Diabetes is a chronic metabolic disease that has become one of the major social medical burdens, affecting approximately 430 million people worldwide. Diabetes can be divided into adult diabetes, childhood and adolescent diabetes, maternal diabetes, elderly diabetes and so on.

Persistent hyperglycemia can also cause vascular disease, microcirculation disorders, and damage to parenchymal circulatory organs, leading to the occurrence and development of other diseases, such as diabetic nephropathy, diabetic lower extremity vascular disease, diabetic foot disease, diabetic retinopathy, diabetic macular degeneration, Diabetic neuropathy, cardiovascular disease, etc.

Although physical exercise and dietary modification have long been recommended to control hyperglycemia, the vast majority of patients still require drug therapy. At present, hypoglycemic drugs mainly include insulin and its analogs, biguanides, sulfonylureas, thiazolidinones, glinides, α-glucosidase inhibitors, glucagon-like peptide-1 (GLP-1) receptors Agonist, dipeptidyl peptidase-4 (DPP-4) inhibitor, sodium-glucose transporter 2 (SGLT2) inhibitor. Different drugs have their own characteristics:

Insulin drugs are an effective means to control hyperglycemia, which can directly reduce blood sugar, such as insulin degludec, insulin glargine, etc., but so far insulin can only be administered by injection, and it relies on complex drug delivery devices to precisely control the dose. May cause risk of hypoglycemia. Biguanides such as metformin are the first-line hypoglycemic drugs and basic drugs recommended by clinical guidelines. They reduce blood sugar by reducing hepatic glucose output and improving peripheral insulin resistance, but the incidence of gastrointestinal adverse reactions is high. Sulfonylureas and glinides can stimulate islet beta cells to secrete insulin and lower blood sugar, such as glibenclamide, glimepiride, gliclazide, glipizide, glipizide, repaglipide Chennai, nateglinide, and mitiglinide, etc. However, these two types of drugs have adverse reactions such as hypoglycemia, weight gain, liver and kidney toxicity. Thiazolidinones, such as rosiglitazone and pioglitazone, lower blood sugar by increasing the sensitivity of target cells to insulin, but weight gain and edema are common adverse effects, and they also increase the risk of fractures and heart failure. Alpha-glucosidase inhibitors reduce postprandial blood sugar by inhibiting the absorption of carbohydrates in the upper small intestine, mainly acarbose, voglibose and miglitol, etc., but gastrointestinal problems such as abdominal distension and flatulence Reactions are very common. Glucagon-like peptide-1 (GLP-1) receptor agonists that inhibit glucagon secretion and increase insulin secretion, such as exenatide, liraglutide, lixisenatide, and benaglutide etc., because it acts in a glucose concentration-dependent manner, it does not significantly increase the risk of hypoglycemia, but it also requires injection, and there are common adverse reactions in the gastrointestinal tract such as nausea and vomiting. Dipeptidyl peptidase-4 (DPP-4) inhibitors lower blood sugar by inhibiting DPP-4, reducing GLP-1 inactivation, enhancing insulin secretion, such as sitagliptin, saxagliptin, vildagliptin, Linagliptin and alogliptin, etc., but they can cause mild to moderate weight gain, and those with liver and kidney insufficiency may need to adjust the dose. Sodium-glucose transporter 2 (SGLT2) inhibitors reduce blood sugar by inhibiting the reabsorption of glucose in the renal tubules and promoting the excretion of glucose in urine, such as dapagliflozin, empagliflozin and canagliflozin. Reactions include genitourinary tract infections, and use is limited in patients with renal impairment.

In current medical practice, hyperglycemia cannot be completely cured, but requires long-term medication. It can be seen from the situation of existing hypoglycemic drugs that all existing drugs have adverse reactions to varying degrees, which has become a challenge for patients who take long-term medication, and even become the main reason for drug discontinuation. Therefore, how to provide a safer and more effective hypoglycemic drug is one of the directions worthy of research at present.

SUMMARY OF THE INVENTION

One of the objects of the present invention is to provide a set of applications. After in-depth research, the inventor provides the following technical solutions.

Application of a combination comprising lysozyme and dosima ester in the preparation of a medicament for lowering blood sugar.

Application of a combination comprising lysozyme and dosima ester in the preparation of a medicament for preventing or treating diseases or symptoms related to hyperglycemia.

In some instances, the disease or condition associated with hyperglycemia can be selected from diabetes, diabetic nephropathy, diabetic lower extremity vascular disease, diabetic foot disease, diabetic retinopathy, diabetic macular degeneration, diabetic neuropathy, cardiovascular disease caused by hyperglycemia disease, etc. Said diabetes is preferably type 2 diabetes.

In some instances, the combination comprising lysozyme and dosima may also not cause gastrointestinal adverse reactions, eg, not cause digestive dysfunction, such as diarrhea, constipation, nausea, bloating, abdominal pain, vomiting, and other symptoms.

In some examples, the combination comprising lysozyme and dosimamate can also improve gastrointestinal diseases or symptoms, eg, improve digestive disorders such as diarrhea, constipation, nausea, bloating, abdominal pain, vomiting, and the like. In some instances, the gastrointestinal disease or symptom is due to hyperglycemia or diabetes.

In some instances, the combination comprising lysozyme and dosimamate can also enhance the effect of other hypoglycemic drugs.

In some instances, the combination comprising lysozyme and dosemamate can also alleviate adverse effects of other antidiabetic drugs.

The application of the combination comprising lysozyme and dosima ester in the preparation of a hypoglycemic drug synergist.

Application of a combination comprising lysozyme and dosima ester in the preparation of a medicine for alleviating adverse reactions of hypoglycemic drugs.

In some examples of the application of synergists and the application of reducing adverse reactions, the hypoglycemic drugs may be selected from insulin and its analogs, biguanide hypoglycemic drugs, sulfonylurea hypoglycemic drugs, thiazolidinone hypoglycemic drugs, Glinide-type hypoglycemic agents, α-glucosidase inhibitor-type hypoglycemic agents, glucagon-like peptide-1 (GLP-1) receptor agonist hypoglycemic agents, dipeptidyl peptidase-4 (DPP) -4) Inhibitor-type hypoglycemic agents, sodium-glucose transporter 2 (SGLT2) inhibitor-type hypoglycemic agents, sodium-glucose transporter 1/2 (SGLT1/2) dual inhibitor-type hypoglycemic agents, or other Drugs with hypoglycemic effect.

In some examples of applications for alleviating adverse reactions, the adverse reactions may be selected from gastrointestinal adverse reactions, hepatic adverse reactions, renal adverse reactions. Gastrointestinal adverse reactions are preferred.

In some examples of the above-mentioned applications, lysozyme and dosimamate can be combined in various ways, including in separate formulations (combination packaging, combination medication), or in the same formulation Combination (combination preparation). The specific formulation form is not particularly limited, and can be various formulations suitable for pharmaceutical use, such as oral formulations, injections, inhalation formulations, and the like.

In some examples of the above several applications, the weight ratio of lysozyme to dosima ester may be 1:100 to 100:1, preferably 1:20 to 20:1, more preferably 1:10 to 10:1.

Another object of the present invention is to provide a set of pharmaceutical compositions.

A pharmaceutical composition containing lysozyme and dosima ester.

In some instances, the pharmaceutical composition is used to lower blood sugar.

In some instances, the pharmaceutical composition is used to prevent or treat a disease or condition associated with hyperglycemia.

In some instances, the disease or condition associated with hyperglycemia can be selected from diabetes, diabetic nephropathy, diabetic lower extremity vascular disease, diabetic foot disease, diabetic retinopathy, diabetic macular degeneration, diabetic neuropathy, cardiovascular disease caused by hyperglycemia disease, etc. Said diabetes is preferably type 2 diabetes.

In some instances, the pharmaceutical composition is used to enhance the efficacy of other antidiabetic drugs.

In some instances, the pharmaceutical composition is used to alleviate other adverse effects of hypoglycemic drugs.

In some examples, in addition to lysozyme and dosimamate, the pharmaceutical composition may also contain other hypoglycemic drugs.

In some instances, the other hypoglycemic drugs can be selected from insulin and its analogs, biguanide hypoglycemic drugs, sulfonylurea hypoglycemic drugs, thiazolidinone hypoglycemic drugs, glinides hypoglycemic drugs, Alpha-glucosidase inhibitor class of hypoglycemic drugs, glucagon-like peptide-1 (GLP-1) receptor agonist class of hypoglycemic drugs, dipeptidyl peptidase-4 (DPP-4) inhibitor class of hypoglycemic drugs Drugs, sodium-glucose transporter 2 (SGLT2) inhibitor class hypoglycemic drugs, sodium-glucose transporter 1/2 (SGLT1/2) dual inhibitor class hypoglycemic drugs, or other drugs with hypoglycemic effect.

In some instances, the adverse reactions may be selected from gastrointestinal adverse reactions, hepatic adverse reactions, and renal adverse reactions. Gastrointestinal adverse reactions are preferred.

In some examples of the above-mentioned pharmaceutical compositions, the pharmaceutical compositions may be formulations with different administration routes, or may be formulations with different physical forms. Formulations of different administration routes, such as oral formulations, injections, inhalation formulations, etc., are preferably oral formulations. Formulations in different physical forms, such as solid preparations, semi-solid preparations, liquid preparations, etc., can be specifically tablets (plain tablets, coated tablets, chewable tablets, dispersible tablets, orally disintegrating tablets, buccal tablets, sublingual tablets, bilayer tablets, etc.) tablets, multi-layer tablets), capsules (hard capsules, soft capsules), pills (plain pills, small pills, micropellets, drop pills), powders, granules, solutions, suspensions, and the like.

In some examples of the above-mentioned pharmaceutical compositions, the pharmaceutical compositions may also contain pharmaceutically suitable excipients. Specific excipients need to be compatible with specific formulation forms, including but not limited to diluents, fillers, excipients, lubricants, disintegrants, binders, glidants, absorption enhancers, solvents, preservatives, Solubilizers, adsorbents, flavoring agents, other carriers, coatings, other inert ingredients, or any combination thereof.

In some examples of the above-mentioned pharmaceutical compositions, the pharmaceutical compositions may be formulations in different release forms, such as normal release formulations, delayed release formulations, sustained release formulations or controlled release formulations, and the like. The lysozyme component and/or the dosimamate component of the pharmaceutical composition may be present in different release forms. For example, it can exist in the form of sustained release, can exist in the form of delayed release, can exist in the form of extended release, can exist in the form of controlled release, can exist in the form of enteric release in the form of a gastric release, or a combination of a gastric release and an enteric release. The lysozyme component and/or the dosima ester component can exist in the same release form or in different release forms, for example, lysozyme is present in an enteric release form and dosimamate in a gastric release form .

In some examples of several of the above-mentioned pharmaceutical compositions, the lysozyme component and the dosima ester component may be present in different combinations.

In some examples of the above-mentioned pharmaceutical compositions, dosima ester and lysozyme are present in a physically combined manner, for example, the two components are mixed together and then prepared into a specific formulation.

In some examples of the above-mentioned pharmaceutical compositions, dosima ester and lysozyme may also exist in a physically separated manner, for example, the two components are separately treated with excipients, for example, the same or different dosage forms are first prepared separately. After the unit, it is then prepared to obtain the final dosage form. The dosage form subunits can be dosage form subunits of various physical dosage forms, such as granules, capsules, powders, tablets, etc.; dosage form subunits can also be dosage form subunits of various release forms, such as delayed release, sustained release, controlled release, Enteric-coated, gastric-coated, gastric-coated and enteric-coated, etc. Dosima ester and lysozyme can be present in the same form of formulation subunit or in different forms of formulation subunit. In some examples, lysozyme is prepared as a formulation subunit in enteric form, and dosimamate is prepared as a formulation subunit in enteric form. In some examples, lysozyme is prepared as a formulation subunit in enteric form and dosimamate is prepared as a formulation subunit in gastric form. For example, lysozyme is prepared into enteric-coated granules, enteric-coated pellets or enteric-coated tablets, dosima ester is prepared into granules and small tablets, and the preparation subunits are assembled into final preparations, such as capsules, tablets, granules, and the like.

In some examples of the above-mentioned pharmaceutical compositions, the weight ratio of lysozyme and dosima ester may be 1:100 to 100:1, preferably 1:20 to 20:1, more preferably 1:10 to 10:1: 1.

The third object of the present invention is to provide a group of combined drugs.

A combination drug that includes lysozyme and dosimamate.

In some examples, lysozyme and dosimamate are used in combination.

In some examples, the lysozyme is in a separate formulation and the doximab is in a separate formulation. There can be various specific physical forms, release forms, auxiliary materials, etc. of the preparation, as described in the technical scheme of the pharmaceutical composition.

In some instances, the combination drug has one or more effects of lowering blood sugar, preventing or treating diseases or symptoms related to hyperglycemia, enhancing the efficacy of other hypoglycemic drugs, or reducing adverse reactions of other hypoglycemic drugs . The diseases or symptoms related to hyperglycemia are as described in the technical scheme of the pharmaceutical composition. The other hypoglycemic drugs are as described in the technical scheme of the pharmaceutical composition.

In some examples, the weight ratio of lysozyme and dosima ester may be 1:100 to 100:1, preferably 1:20 to 20:1, more preferably 1:10 to 10:1.

A fourth object of the present invention is to provide a set of methods.

A method of lowering blood sugar comprising administering to a subject in need thereof an effective amount of lysozyme and dosima ester.

A method of preventing or treating a disease or condition associated with hyperglycemia, comprising administering to a subject in need thereof an effective amount of lysozyme and doximab.

In some instances, the disease or condition associated with hyperglycemia can be selected from diabetes, diabetic nephropathy, diabetic lower extremity vascular disease, diabetic foot disease, diabetic retinopathy, diabetic macular degeneration, diabetic neuropathy, cardiovascular disease caused by hyperglycemia disease, etc. Said diabetes is preferably type 2 diabetes.

In some instances, the subject's blood glucose level is higher than normal.

In some instances, the subject also has co-morbidities or symptoms of the gastrointestinal tract, such as gastrointestinal dysfunction, eg, diarrhea, constipation, nausea, bloating, abdominal pain, vomiting, and the like. In some instances, the gastrointestinal disease or symptom is due to hyperglycemia or diabetes.

A method of increasing the efficacy of a hypoglycemic drug, comprising combining lysozyme and dosimamate with the hypoglycemic drug.

A method for alleviating adverse effects of a hypoglycemic drug, comprising combining lysozyme and doximab with the hypoglycemic drug.

In some examples of the method of increasing the efficacy of the hypoglycemic drug and the method of reducing the adverse reaction of the hypoglycemic drug, the hypoglycemic drug may be selected from insulin and its analogs, biguanide hypoglycemic drugs, sulfonylurea hypoglycemic drugs, thiazolidinone Antidiabetic drugs, glinide antidiabetic drugs, α-glucosidase inhibitor antidiabetic drugs, glucagon-like peptide-1 (GLP-1) receptor agonist antidiabetic drugs, dipeptidyl peptides Enzyme-4 (DPP-4) inhibitor class hypoglycemic drugs, sodium-glucose transporter 2 (SGLT2) inhibitor class hypoglycemic drugs, sodium-glucose transporter 1/2 (SGLT1/2) dual inhibitor class hypoglycemic drugs medicines, or other medicines that have a blood sugar-lowering effect.

In some examples of the method of increasing the efficacy of a hypoglycemic drug and the method of reducing adverse effects of a hypoglycemic drug, lysozyme and dosemalate are administered before, after, or concurrently with the administration of the hypoglycemic drug.

In some examples of the method for alleviating adverse reactions of hypoglycemic drugs, the adverse reactions may be selected from gastrointestinal adverse reactions, liver adverse reactions, and renal adverse reactions. Gastrointestinal adverse reactions are preferred.

In some examples of the several methods described above, lysozyme and dosimamate can be combined in a variety of ways, including in separate formulations (packaging, combination), or in the same formulation Combination (combination preparation). The specific formulation form is not particularly limited, and can be various formulations suitable for pharmaceutical use, such as oral formulations, injections, inhalation formulations, and the like.

In some examples of the method of increasing the efficacy of the hypoglycemic drug and the method of reducing the adverse reaction of the hypoglycemic drug, the hypoglycemic drug may be combined with lysozyme and dosemamate in different forms, including in separate formulations. (combination packaging, combination medication), or a combination (combination preparation) may be combined so as to be present in the same preparation.

In some examples of the above several methods, the weight ratio of lysozyme to dosima ester may be 1:100 to 100:1, preferably 1:20 to 20:1, more preferably 1:10 to 10:1.

In some examples of the above-mentioned several methods, the daily dosage of dosmamate is 0.2～20g; preferably, the daily dosage of dosemamate is 1～20g; ~10g.

In some examples of the above methods, the daily dosage of lysozyme is 0.2-20 g; preferably, the daily dosage of lysozyme is 0.5-20 g; more preferably, the daily dosage of lysozyme is 1-10 g .

In some examples of the above several methods, dosima ester and lysozyme can be used 1-3 times/day, respectively.

Beneficial effects:

The inventors have found through research that lysozyme and dosima ester have a significant synergistic effect in reducing blood sugar, and can improve gastrointestinal dysfunction in patients with hyperglycemia. In addition, when lysozyme and dosimamate are used in combination with other hypoglycemic drugs, they can enhance the effect of these hypoglycemic drugs and reduce their adverse reactions. Therefore, the combination of lysozyme and dosima ester is of particular clinical value for hypoglycemia and hyperglycemia-related diseases or symptoms.

Detailed ways

definition:

Blood Glucose: Blood glucose generally refers to the free glucose in the plasma. Glucose also combines with hemoglobin in blood to form glycated hemoglobin. The ratio of glycated hemoglobin is generally proportional to the concentration of free glucose in plasma over a period of time, so it can also be used to evaluate blood sugar levels.

Hyperglycemia: Under normal circumstances, the blood sugar concentration of the human body is generally kept constant within the range of 4-7mmol/L. Anything above the normal range is called hyperglycemia.

Diabetes: Diabetes is a metabolic disease with persistent hyperglycemia as one of its basic features. Clinically, diabetes can be identified by standard medical diagnostic methods.

Lysozyme: Lysozyme, or lysozyme, is a lysozyme derived from animals, plants, microorganisms, or a recombinant of natural lysozyme. For example, egg lysozyme, human lysozyme, recombinant human lysozyme, phage lysozyme and the like can be used. The lysozyme in the present invention also includes its pharmaceutically acceptable salts, such as hydrochloride, chloride, sulfate or amino acid salt and the like. Lysozyme has antibacterial, antiviral and other activities, and is recognized as a safe substance (GRAS) by the FDA in the United States.

Lysozyme is an endogenous substance in human body, belongs to antimicrobial peptide, and is an indispensable factor for human innate immunity. Lysozyme has also been considered as a biomarker of inflammatory response because lysozyme is distributed in immune cells such as macrophages and neutrophils. There are few reports on the relationship between lysozyme and blood sugar in the existing research.

A recent study found that lysozyme can promote intestinal bacteria to release a Nod1 ligand, which can further promote the efficiency of insulin transport from pancreatic β cells (Zhang, 2019). This study provides a preliminary possibility for the treatment of hyperglycemia with lysozyme. However, the specific effect of lysozyme still needs to be confirmed. We found through research that although lysozyme has a certain effect, the hypoglycemic effect is still not ideal.

By combining another drug, dosima ester, with lysozyme, we unexpectedly found that the hypoglycemic effect was significantly enhanced. It is also found through further research that this combination has a good application prospect, not only has a hypoglycemic effect by itself, but also can be used in combination with the existing hypoglycemic drugs currently on the market to improve the effectiveness and safety of the existing drugs.

Dosima ester: its chemical name is diomin heptabis(bisulfate) aluminum complex, and its molecular formula is Al ₇ (OH) ₁₄ (C ₂₈ H ₂₅ O ₃₆ S ₇ )[Al(OH) ₃ ] ₇ . Dosemamate is a gastric mucosal protective agent that has been marketed for many years for the treatment of gastric and duodenal ulcers.

The present invention will be explained below with specific embodiments. The lysozyme and dosima ester used therein were provided by Xiangbei Wellman Pharmaceutical Co., Ltd.

Example 1 Preparation of lysozyme dosima ester composite capsules

Take 50 g of lysozyme raw material, add 200 g of lactose and 100 g of microcrystalline cellulose, use 5% polyvinylpyrrolidone ethanol solution as a binder, granulate, and dry to obtain lysozyme granules. Take 500 g of dosima ester raw material, 100 g of lactose, 50 g of low-substituted hydroxypropyl cellulose, 2 g of micropowder silica gel, use 5% polyvinylpyrrolidone ethanol solution as a binder, granulate, and dry to obtain dosima ester granules. The lysozyme granules and the dosima ester granules are uniformly mixed and encapsulated to obtain the lysozyme dosima ester composite capsules.

Example 2 Preparation of lysozyme dosima ester enteric-coated composite granules

Take 20 g of lysozyme raw material, add 50 g of lactose, 50 g of pregelatinized starch and 100 g of microcrystalline cellulose, use 5% polyvinylpyrrolidone ethanol solution as a binder, granulate, apply enteric coating, and dry to obtain lysozyme enteric-coated granules . Take 500 g of dosima ester raw material, 200 g of lactose, 100 g of low-substituted hydroxypropyl cellulose, use 5% polyvinylpyrrolidone ethanol solution as a binder, granulate, and dry to obtain dosima ester granules. The lysozyme enteric-coated granules and the dosima ester granules are respectively taken, mixed and bagged to obtain the lysozyme dosima ester enteric-coated composite granules.

Example 3 The effect of the combination of lysozyme and dosima ester on diabetic model animals

Animals: Select 10-week-old clean-grade db/db transgenic mice with fasting blood glucose not lower than 18mmol/L, weighing 40-50g. Adaptive feeding for one week, free access to water, standard diet feeding.

Drugs: lysozyme (provided by Xiangbei Wellman Pharmaceutical Co., Ltd.), dosima ester (provided by Xiangbei Wellman Pharmaceutical Co., Ltd.), sucralfate (commercially available).

Grouping: Animals were randomly divided into model group, lysozyme group, dosemamate group, sucralfate group, lysozyme dosemalate low-dose group, lysozyme dosemamate medium-dose group, lysozyme-dosemalate high-dose group, Lysozyme sucralfate low-dose group, lysozyme sucralfate medium-dose group, and lysozyme sucralfate high-dose group, with 8 animals in each group.

Administration: The model group was given 5% methylcellulose sodium by gavage. Lysozyme group (60mg/kg), dosemamate group (600mg/kg), sucralfate group (600mg/kg), lysozyme dosemamate low-dose group (lysozyme 60mg/kg + dosemamate 6mg/kg) , lysozyme dosima ester medium-dose group (lysozyme 60mg/kg + dosima ester 60mg/kg), lysozyme dosima ester high-dose group (lysozyme 60mg/kg + dosemamate 600mg/kg), lysozyme sucralfate Low-dose group (lysozyme 60mg/kg+sucralfate 6mg/kg), lysozyme-sucralfate medium-dose group (lysozyme 60mg/kg+sucralfate 60mg/kg), lysozyme-sucralfate high-dose group (lysozyme sucralfate 60mg/kg) 60mg/kg + sucralfate 600mg/kg) were given corresponding drugs (mixed with 5% methylcellulose solution) by gavage respectively. It is administered once a day for a period of 4 weeks.

Detection: After the administration was completed, fasting for 8 hours, blood was taken from the tail tip to detect the fasting blood glucose value of the animals in each group.

Results: The fasting blood glucose values of animals in each group are shown in Table 1.

Table 1 Measurement of fasting blood glucose of animals in each group

group number of animals Fasting blood glucose (mmol/L) model group 8 24.33±2.80 Lysozyme group 8 22.20±4.29 Dosima ester group 8 23.65±3.84 Sucralfate group 8 23.36±3.01 Lysozyme dosima ester low-dose group 8 20.11±2.57* Lysozyme dosima ester medium dose group 8 19.28±2.05**

High-dose group of lysozyme dosima ester 8 17.84±2.65** Lysozyme sucralfate low-dose group 8 22.11±2.08 Lysozyme sucralfate middle dose group 8 22.71±1.44 Lysozyme sucralfate high-dose group 8 21.31±1.67

Note: p < 0.05 (*), p < 0.01 (**) compared with the model group.

Conclusion: db/db mice are characterized by overeating, obesity, hyperglycemia and insulin resistance due to leptin gene defect, and are a common animal model of spontaneous diabetes. In this experiment, the short-term hypoglycemic effects of lysozyme, dosimamate, sucralfate, and several drug combinations were explorably studied, and it was found that lysozyme could only reduce blood sugar slightly, but there was no significant difference compared with the model group. The combination of sima ester can greatly enhance the hypoglycemic effect, with a very significant difference compared with the model group, while the assisted hypoglycemic effect of sucralfate is almost negligible.

Example 4 Long-term effects of lysozyme and dosima ester combined with other hypoglycemic drugs on diabetic model animals

Animals: 10-week-old clean-grade db/db transgenic mice with fasting blood glucose not lower than 18mmol/L, weighing 40-50g, and 10-week-old littermate wild-type C57BL/6J mice, weighing 20-25g. Animals were adaptively fed for one week, with free access to water and standard chow.

Drugs: lysozyme granules (Example 1), lysozyme enteric-coated granules (Example 2), dosima ester granules (Example 1), acarbose tablets (commercially available), and metformin hydrochloride tablets (commercially available).

Grouping: C57BL/6J mice were set as normal group. The db/db transgenic mice were randomly divided into model group, metformin group, acarbose group, lysozyme dosmarate group, lysozyme dosmarate metformin group, lysozyme dosmarate acarbose group, lysozyme dosemamate group Sima ester enteric-coated group. Each group consisted of 10 animals.

Administration: Normal group and model group were given normal saline by gavage. Metformin group (250mg/kg), acarbose group (50mg/kg), lysozyme dosemalate group (lysozyme 100mg/kg + dosemamate 300mg/kg), lysozyme dosemamate metformin group (lysozyme 100mg /kg + dosemamate 300mg/kg + metformin 250mg/kg), lysozyme dosemamate acarbose group (lysozyme 100mg/kg + dosmamate 300mg/kg + acarbose 50mg/kg), lysozyme dosemamate Enteric-coated group (lysozyme 100mg/kg + dosima ester 300mg/kg) was given the corresponding medicines by gavage respectively according to the dose (calculated as active ingredient), and the lysozyme dosima ester enteric-coated group used lysozyme enteric-coated granules, other lysozyme-containing groups used lysozyme granules. It was administered once a day for 10 weeks.

Blood glucose measurement: At the 5th week after administration and after administration (10th week), the fasting blood glucose values of animals in each group were measured.

Determination of gastric emptying rate: animals in each group were fasted for 6 hours after the completion of blood glucose measurement, gavaged with 2 ml of 1.5 mmol/L phenol red solution, laparotomy after anesthesia, ligated the pylorus and cardia, and removed the stomach. Cut open, rinse the inside of the stomach with distilled water, dilute the stomach contents to 20ml, add 20ml of 0.5mol/L sodium hydroxide solution, mix well, and let stand. 5 ml of the supernatant was taken, 0.5 ml of 20% trichloroacetic acid was added, and centrifuged to serve as the assay solution. The absorbance at the wavelength of 560 nm was measured with a microplate reader, and the gastric emptying rate was calculated. Gastric emptying rate=(1-measurement solution absorbance/standard phenol red solution absorbance)×100%.

Determination of small intestine advancement rate: Take out the small intestine, lay it on paper, measure the distance from the pylorus to the ileum (the full length of the small intestine), and the distance from the pylorus to the farthest distance from phenol red (the moving distance of phenol red), calculate the small intestine advancement rate . Small intestine advancement rate=(phenol red movement distance/small intestine total length)×100%.

Results: The main results of this experiment are shown in Table 2 and Table 3.

Table 2 Fasting blood glucose measurement of animals in each group

Note: p < 0.05 (*), p < 0.01 (**) compared with the model group.

Table 3 Gastric emptying rate and small intestinal propulsion rate of animals in each group

Note: p < 0.05 (*), p < 0.01 (**) compared with the model group.

Conclusion: This experiment studies the long-term hypoglycemic effect of lysozyme dosima ester combination on diabetic model mice, and compares it with the positive drugs metformin and acarbose, and also conducts a three-drug combination study. It was found that the combination of lysozyme dosima ester has a long-term hypoglycemic effect, and the combination of lysozyme dosima ester can also enhance the hypoglycemic effect of metformin or acarbose, and the enteric-coated preparation has better effect. Metformin is a typical guanidine hypoglycemic drug, acarbose is a typical α-glucosidase inhibitor, and they are very commonly used clinical hypoglycemic drugs. The combination of the present invention shows a synergistic potential to existing hypoglycemic drugs.

The effects of several drugs on gastrointestinal function were also investigated in this experiment. It was found that db/db mice developed more severe digestive dysfunction (such as decreased gastric emptying rate and small intestinal propulsion rate) around 20 weeks, but existing hypoglycemic drugs such as metformin or acarbose could only improve digestive function. deterioration. However, unexpectedly, the combination of lysozyme dosima ester of the present invention can significantly increase gastric emptying rate and small intestinal propulsion rate, that is, significantly improve the digestive dysfunction of diabetic model animals, especially the enteric-coated preparation has better effect. In view of the wide range of gastrointestinal adverse reactions such as digestive dysfunction, such as diarrhea, constipation, nausea, abdominal distension, abdominal pain, vomiting, etc., in existing hypoglycemic drugs, it is reported that 20% of diabetic patients using metformin clinically have gastric Therefore, the combination of the present invention has high clinical value in improving the compliance and tolerance of diabetic patients.

The present invention has been described in detail above. On this basis, those skilled in the art can make some changes or improvements to it. These changes or improvements made without departing from the spirit of the present invention all belong to the requirements of the present invention. protected content.

Claims (36)

Application of a combination comprising lysozyme and dosima ester in the preparation of a medicament for lowering blood sugar. Application of a combination comprising lysozyme and dosima ester in the preparation of a medicament for preventing or treating diseases or symptoms related to hyperglycemia. The use according to claim 2, wherein the disease or symptom related to hyperglycemia is selected from diabetes, diabetic nephropathy, diabetic lower extremity vascular disease, diabetic foot disease, diabetic retinopathy, diabetic macular degeneration, diabetic neuropathy or hyperglycemia one or more of cardiovascular diseases. The application of the combination comprising lysozyme and dosima ester in the preparation of a drug for enhancing the efficacy of a hypoglycemic drug and/or alleviating the adverse reaction of the hypoglycemic drug. The application according to claim 4, wherein the hypoglycemic drug is selected from insulin and its analogs, biguanide hypoglycemic drugs, sulfonylurea hypoglycemic drugs, thiazolidinone hypoglycemic drugs, glinide hypoglycemic drugs Drugs, α-glucosidase inhibitor class hypoglycemic drugs, glucagon-like peptide-1 receptor agonist class hypoglycemic drugs, dipeptidyl peptidase-4 inhibitor class hypoglycemic drugs, sodium-glucose transporter 2 Inhibitor-type hypoglycemic agents, or one or more of the sodium-glucose transporter 1/2 dual inhibitor-type hypoglycemic agents. The application according to claim 4, wherein the adverse reactions are selected from gastrointestinal adverse reactions, liver adverse reactions, or renal adverse reactions. A pharmaceutical composition containing lysozyme and dosima ester. The pharmaceutical composition according to claim 7, wherein the weight ratio of lysozyme and dosima ester in the pharmaceutical composition is 1:100 to 100:1. The pharmaceutical composition according to claim 8, wherein the weight ratio of lysozyme and dosima ester in the pharmaceutical composition is 1:20 to 20:1. The pharmaceutical composition according to claim 8, wherein the weight ratio of lysozyme and dosima ester in the pharmaceutical composition is 1:10 to 10:1. The pharmaceutical composition according to claim 7, further comprising other hypoglycemic drugs. The pharmaceutical composition according to claim 11, wherein the other hypoglycemic drugs are selected from insulin and its analogs, biguanide hypoglycemic drugs, sulfonylurea hypoglycemic drugs, thiazolidinone hypoglycemic drugs, glinide Hypoglycemic drugs, α-glucosidase inhibitor-based hypoglycemic drugs, glucagon-like peptide-1 receptor agonist hypoglycemic drugs, dipeptidyl peptidase-4 inhibitor-based hypoglycemic drugs, sodium-glucose Transporter 2 inhibitor class of hypoglycemic agents, or one or more of the sodium-glucose transporter 1/2 dual inhibitor class of hypoglycemic drugs. The pharmaceutical composition according to claim 7, further comprising pharmaceutically applicable excipients. The pharmaceutical composition according to claim 7, which is an oral preparation, an injection preparation or an inhalation preparation. The pharmaceutical composition according to claim 7, which is a solid preparation, a semi-solid preparation or a liquid preparation. The pharmaceutical composition according to claim 7, which is a normal-release preparation, a delayed-release preparation, a sustained-release preparation or a controlled-release preparation. According to the pharmaceutical composition of claim 7, the release forms of lysozyme and dosemaester in the pharmaceutical composition can be the same or different. The pharmaceutical composition according to claim 17, wherein the dosimamate in the pharmaceutical composition is released by gastric dissolution. The pharmaceutical composition according to claim 17, wherein the lysozyme in the pharmaceutical composition is enteric-coated release. A combination drug that includes lysozyme and dosimamate. The combination medicine according to claim 20, wherein the lysozyme in the combination medicine is in the form of an independent preparation, and the dosima ester is in the form of an independent preparation. The drug combination according to claim 21, wherein the lysozyme and/or dosemamate are oral preparations, injection preparations or inhalation preparations. The drug combination according to claim 21, wherein the lysozyme and/or dosemaester is a solid preparation, a semi-solid preparation or a liquid preparation. The drug combination according to claim 21, wherein the lysozyme and/or dosima ester is a normal-release preparation, a delayed-release preparation, a sustained-release preparation or a controlled-release preparation. The combination medicine according to claim 20, wherein the weight ratio of lysozyme and dosima ester in the combination medicine is 1:100 to 100:1. The combined drug according to claim 20, further comprising other hypoglycemic drugs. The combination drug according to claim 26, wherein the other hypoglycemic drugs are selected from insulin and its analogs, biguanide hypoglycemic drugs, sulfonylurea hypoglycemic drugs, thiazolidinone hypoglycemic drugs, glinide Hypoglycemic drugs, α-glucosidase inhibitor-based hypoglycemic drugs, glucagon-like peptide-1 receptor agonist hypoglycemic drugs, dipeptidyl peptidase-4 inhibitor-based hypoglycemic drugs, sodium-glucose Transporter 2 inhibitor class of hypoglycemic agents, or one or more of the sodium-glucose transporter 1/2 dual inhibitor class of hypoglycemic drugs. A method of lowering blood sugar comprising administering to a subject in need thereof an effective amount of lysozyme and dosima ester. A method of preventing or treating a disease or condition associated with hyperglycemia, comprising administering to a subject in need thereof an effective amount of lysozyme and dosima ester. The method of claim 29, wherein the hyperglycemia-related disease or condition is selected from the group consisting of diabetes, diabetic nephropathy, diabetic lower extremity vasculopathy, diabetic foot disease, diabetic retinopathy, diabetic macular degeneration, diabetic neuropathy or hyperglycemia lead to cardiovascular disease. A method for enhancing the efficacy of a hypoglycemic drug, comprising combining lysozyme and dosima ester with the hypoglycemic drug. A method for alleviating adverse effects of a hypoglycemic drug, comprising combining lysozyme and doximab with the hypoglycemic drug. The method according to claim 32, wherein the adverse reactions are selected from one or more of gastrointestinal adverse reactions, liver adverse reactions, and renal adverse reactions. According to the method of any one of claims 28-33, the weight ratio of the amount of lysozyme and dosima ester is 1:100 to 100:1. According to the method of any one of claims 28-33, the daily dosage of the dosima ester is 0.2-20 g. According to the method of any one of claims 28-33, the daily dosage of the lysozyme is 0.2-20 g.