CN116157132A

CN116157132A - Combination, application and method for reducing blood sugar

Info

Publication number: CN116157132A
Application number: CN202180052825.6A
Authority: CN
Inventors: 孙天宇; 孙明杰; 蒋娟艳; 高太平
Original assignee: Guangzhou Xinchuangyi Biomedical Co ltd; Guangzhou Xinchuangyi Co ltd; Guangzhou Xinchuangyi Medicine Clinical Research Co ltd; Nanjing Kangfushun Pharmaceutical Co Ltd; Xiangbei Welman Pharmaceutical Co Ltd
Current assignee: Guangzhou Xinchuangyi Biomedical Co ltd; Guangzhou Xinchuangyi Co ltd; Guangzhou Xinchuangyi Medicine Clinical Research Co ltd; Nanjing Kangfushun Pharmaceutical Co Ltd; Xiangbei Welman Pharmaceutical Co Ltd
Priority date: 2020-10-30
Filing date: 2021-09-28
Publication date: 2023-05-23
Also published as: WO2022089133A1

Abstract

The present invention provides a combination for lowering blood glucose comprising lysozyme and dossmate. The invention also provides applications and methods thereof. Lysozyme and dossmate have a significant synergistic effect in lowering blood glucose and can improve gastrointestinal dysfunction in hyperglycemic patients. In addition, the combination of the invention has synergistic and attenuation effects on other hypoglycemic drugs, so that the combination is particularly of clinical value for reducing blood sugar and hyperglycemia-related diseases or symptoms.

Description

Combination, application and method for reducing blood sugar

Technical Field

The invention belongs to the field of medicines, and particularly relates to a combination, application and method for reducing blood sugar.

Background

Glucose is the main energy source for normal physiological activities of the human body. The food is partially converted into glucose after being digested by gastrointestinal tract, absorbed into blood, and supplied to various organs and tissues of the whole body through blood circulation. Maintaining a certain blood glucose level is critical for the human body to perform physiological functions. Typically, the blood glucose concentration in humans will be maintained at a relatively constant level, with blood glucose levels (FPG) of about 4-7mmol/L in the fasting state in adults. And hyperglycemia is a common pathological condition.

Fasting blood glucose above 6mmol/L reflects a disorder of glucose metabolism, known as prediabetes.

Diabetes can be diagnosed by having fasting blood glucose of greater than 7mmol/L or glycosylated hemoglobin (HbA 1 c) of greater than 6.5%. Diabetes is a chronic metabolic disease, which has become one of the major social medical burden, affecting about 4.3 million people worldwide. Diabetes mellitus is classified into adult diabetes mellitus, childhood and adolescent diabetes mellitus, pregnant woman diabetes mellitus, senile diabetes mellitus and the like according to the population.

Persistent hyperglycemia also causes vascular lesions, microcirculation disorders, damage to the parenchymal circulatory organs, etc., leading to the development and progression of other diseases such as diabetic nephropathy, diabetic lower limb vascular lesions, diabetic foot disease, diabetic retinopathy, diabetic macular degeneration, diabetic neuropathy, cardiovascular disease, etc.

While physical exercise and dietary regulation have long been recommended for controlling hyperglycemia, a vast majority of patients still need medication. Currently, hypoglycemic agents are mainly insulin and its analogues, biguanides, sulfonylureas, thiazolidinediones, glinide, alpha-glucosidase inhibitors, glucagon-like peptide-1 (GLP-1) receptor agonists, dipeptidyl peptidase-4 (DPP-4) inhibitors, sodium-glucose transporter 2 (SGLT 2) inhibitors. The different medicines have the characteristics that:

insulin-like drugs are effective means for controlling hyperglycemia, and can directly lower blood sugar, such as insulin deluge, insulin glargine, etc., but insulin has so far been administered only by injection and relies on sophisticated administration devices to precisely control the dosage, insulin may lead to the risk of hypoglycemia. Biguanides such as metformin are first-line hypoglycemic drugs and basic drugs recommended by clinical guidelines, which lower blood glucose by reducing hepatic glucose output and improving peripheral insulin resistance, but have a higher incidence of gastrointestinal adverse reactions. Sulfonylureas and glinides stimulate insulin secretion from the beta cells of the islets of langerhans to lower blood glucose, such as glibenclamide, glimepiride, gliclazide, glipizide, gliquidone, repaglinide, nateglinide, mitiglinide, and the like. However, both of these drugs have adverse reactions such as hypoglycemia, weight gain, hepatorenal toxicity, etc. Thiazolidinediones lower blood glucose by increasing the sensitivity of target cells to insulin, such as rosiglitazone and pioglitazone, but weight gain and oedema are common adverse effects thereof, and additionally increase the risk of bone fracture and heart failure. Alpha-glucosidase inhibitors reduce postprandial blood glucose by inhibiting absorption of carbohydrates in the upper part of the small intestine, mainly acarbose, voglibose, miglitol and the like, but gastrointestinal adverse reactions such as abdominal distension, exhaustion and the like are very common. Glucagon-like peptide-1 (GLP-1) receptor agonists inhibit glucagon secretion, increase insulin secretion, such as exenatide, liraglutide, risinatide, benalunin, etc., do not significantly increase the risk of hypoglycemia because they act in a glucose concentration dependent manner, but are also administered by injection and have the common adverse effects of the gastrointestinal tract such as nausea, vomiting, etc. Dipeptidyl peptidase-4 (DPP-4) inhibitors lower blood glucose by inhibiting DPP-4, reducing GLP-1 inactivation, enhancing insulin secretion, such as sitagliptin, saxagliptin, vildagliptin, linagliptin, and alogliptin, etc., but they may slightly increase body weight, and in addition, liver and kidney dysfunction may require adjustment of dosage. Sodium-glucose transporter 2 (SGLT 2) inhibitors reduce blood glucose, such as dapagliflozin, engagliflozin, and canagliflozin, by inhibiting glucose reabsorption in the renal tubules, promoting urinary glucose excretion, and their common adverse effects including genitourinary tract infections, and their use in patients with impaired renal function has been limited.

Hyperglycemia in current medical practice is not completely cured, but rather requires long-term administration. From the situation of the existing hypoglycemic drugs, adverse reactions exist to different degrees in the existing drugs, which becomes a challenge for patients taking the drugs for a long time and even becomes a main cause of drug withdrawal. Therefore, how to provide a safer and more effective hypoglycemic drug is one of the directions of current research.

Disclosure of Invention

It is an object of the present invention to provide a set of applications. The inventors have conducted intensive studies to provide the following technical solutions.

The use of a combination comprising lysozyme and dossmate for the manufacture of a medicament for lowering blood glucose.

Use of a combination comprising lysozyme and dossmate for the manufacture of a medicament for the prevention or treatment of a disease or condition associated with hyperglycemia.

In some examples, the disease or condition associated with hyperglycemia may be selected from diabetes, diabetic nephropathy, diabetic lower limb vascular disease, diabetic foot disease, diabetic retinopathy, diabetic macular degeneration, diabetic neuropathy, cardiovascular disease caused by hyperglycemia, and the like. The diabetes is preferably type 2 diabetes.

In some examples, the combination comprising lysozyme and driller's ester may also not cause adverse gastrointestinal reactions, such as digestive dysfunction, such as diarrhea, constipation, nausea, abdominal distension, abdominal pain, vomiting, and the like.

In some examples, the combination comprising lysozyme and driller's ester may also improve gastrointestinal diseases or symptoms, such as improving digestive dysfunction, such as symptoms of diarrhea, constipation, nausea, abdominal distension, abdominal pain, vomiting, and the like. In some examples, the gastrointestinal disease or condition is due to hyperglycemia or diabetes.

In some examples, the combination comprising lysozyme and dossmate may also enhance the effects of other hypoglycemic agents.

In some examples, the combination comprising lysozyme and driller's ester may also alleviate adverse effects of other hypoglycemic agents.

Comprises the application of the combination of lysozyme and dossmate in preparing the synergist of hypoglycemic drugs.

The use of a combination comprising lysozyme and dossmate in the manufacture of a medicament for reducing adverse effects of a hypoglycemic agent.

In some examples of the use of potentiators and the use of alleviating adverse effects, the hypoglycemic agent may be selected from insulin and analogues thereof, biguanide hypoglycemic agents, sulfonylurea hypoglycemic agents, thiazolidineone hypoglycemic agents, glinide hypoglycemic agents, alpha-glucosidase inhibitor hypoglycemic agents, glucagon-like peptide-1 (GLP-1) receptor agonist hypoglycemic agents, dipeptidylpeptidase-4 (DPP-4) inhibitor hypoglycemic agents, sodium-glucose transporter 2 (SGLT 2) inhibitor hypoglycemic agents, sodium-glucose transporter 1/2 (SGLT 1/2) dual inhibitor hypoglycemic agents, or other agents having hypoglycemic effects.

In some examples of the use to reduce adverse effects, the adverse effects may be selected from gastrointestinal adverse effects, liver adverse effects, kidney adverse effects. Preferably adverse gastrointestinal reactions.

In some examples of the several applications described above, lysozyme and polyram may be combined in a variety of ways, including in separate formulations (combination package, co-administration), or in the presence of the same formulation (co-formulation). The specific formulation is not particularly limited, and may be various dosage forms suitable for pharmaceutical use, such as oral preparations, injections, inhalants, and the like.

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

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

A pharmaceutical composition comprises lysozyme and dossmate.

In some examples, the pharmaceutical composition is for lowering blood glucose.

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

In some examples, the pharmaceutical composition is used to enhance the efficacy of other hypoglycemic agents.

In some examples, the pharmaceutical composition is used to reduce adverse effects of other hypoglycemic agents.

In some examples, the pharmaceutical composition may contain other hypoglycemic agents in addition to lysozyme and dossmate.

In some examples, the additional hypoglycemic agent may be selected from insulin and analogues thereof, biguanide hypoglycemic agents, sulfonylurea hypoglycemic agents, thiazolidineone hypoglycemic agents, glinide hypoglycemic agents, alpha-glucosidase inhibitor hypoglycemic agents, glucagon-like peptide-1 (GLP-1) receptor agonist hypoglycemic agents, dipeptidyl peptidase-4 (DPP-4) inhibitor hypoglycemic agents, sodium-glucose transporter 2 (SGLT 2) inhibitor hypoglycemic agents, sodium-glucose transporter 1/2 (SGLT 1/2) dual inhibitor hypoglycemic agents, or other agents having hypoglycemic effects.

In some examples, the adverse reaction may be selected from gastrointestinal adverse reactions, liver adverse reactions, kidney adverse reactions. Preferably adverse gastrointestinal reactions.

In some examples of the above-described pharmaceutical compositions, the pharmaceutical compositions may be formulated for different routes of administration or may be formulated for different physical forms. Formulations for different routes of administration, such as oral formulations, injections, inhalant formulations, etc., are preferred. The preparations of different physical forms such as solid preparations, semisolid preparations, liquid preparations and the like may be specifically tablets (plain tablets, coated tablets, chewable tablets, dispersible tablets, orally disintegrating tablets, buccal tablets, sublingual tablets, bilayer tablets, multilayer tablets), capsules (hard capsules, soft capsules), pills (ordinary pills, pellets, micropellets, dripping pills), powders, granules, solutions, suspensions and the like.

In some examples of the above pharmaceutical compositions, the pharmaceutical compositions may further comprise pharmaceutically suitable excipients. Specific excipients are required to be compatible with the specific formulation, including, but not limited to, diluents, fillers, excipients, lubricants, disintegrants, binders, glidants, absorption enhancers, solvents, preservatives, solubilizers, adsorbents, flavoring agents, other carriers, coating materials, other inert ingredients, or any combination thereof.

In some examples of the above-mentioned several pharmaceutical compositions, the pharmaceutical compositions may be in different release forms of the formulation, such as a normal release formulation, a delayed release formulation, a sustained release formulation, a controlled release formulation, or the like. The lysozyme component and/or the multi-judicial maleate component of the pharmaceutical composition may be present in different release forms. For example, it may be in the form of a sustained release, it may be in the form of a delayed release, it may be in the form of a controlled release, it may be in the form of an enteric release, it may be in the form of a gastric release, or it may be in the form of a combination of gastric and enteric releases. The lysozyme component and/or the esma ester component may be present in the same release form or may be present in different release forms, e.g. lysozyme in an enteric release form and esma ester in a gastric release form.

In some examples of the several pharmaceutical compositions described above, the lysozyme component and the doesmaleate component may be present in different combinations.

In some examples of the several pharmaceutical compositions described above, the driller's ester and lysozyme are present in a physically combined form, such as by mixing the two components together before preparing them into a particular formulation.

In some examples of the above-described pharmaceutical compositions, the driller's ester and lysozyme may also be present in a physically separate form, e.g., the two components are treated separately with excipients, e.g., after being separately formulated into the same or different dosage form subunits, and then prepared into the final dosage form. The dosage form subunits may be dosage form subunits of various physical dosage forms, such as granules, capsules, powders, tablets, etc.; the dosage form subunits may also be dosage form subunits in various release forms such as delayed release, sustained release, controlled release, enteric, gastric and enteric, etc. The tosylate and lysozyme may exist as the same form of formulation subunit, or may exist as different forms of formulation subunit. In some examples, lysozyme is prepared as an enteric form of the formulation subunit, and dossmate is prepared as an enteric form of the formulation subunit. In some examples, lysozyme is prepared as an enteric form of the formulation subunit and dossmate is prepared as a gastric form of the formulation subunit. For example, lysozyme is prepared into enteric granules, enteric pellets or enteric tablets, dosma ester is prepared into granules, tablets, and the formulation subunits are assembled into final formulations, such as capsules, tablets, granules, and the like.

In some examples of the several pharmaceutical compositions described above, the weight ratio of lysozyme to dosmate may be from 1:100 to 100:1, preferably 1:20 to 20:1, more preferably 1:10 to 10:1.

it is a further object of the present invention to provide a combination.

A combination comprising lysozyme and dossmate.

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

In some examples, the lysozyme is in the form of a stand-alone formulation and the domamate is in the form of a stand-alone formulation. The specific physical form, release form, auxiliary materials and the like of the preparation can be various, and are described in the technical scheme of the pharmaceutical composition.

In some examples, the combination has one or more of lowering blood glucose, preventing or treating a disease or symptom associated with hyperglycemia, enhancing the efficacy of other hypoglycemic agents, or alleviating adverse effects of other hypoglycemic agents. 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 to dosmate may be from 1:100 to 100:1, preferably 1:20 to 20:1, more preferably 1:10 to 10:1.

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

A method of lowering blood glucose comprising administering to a subject in need thereof effective amounts of lysozyme and dossmate.

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 dossmate.

In some examples, the subject has a blood glucose level that is above normal.

In some examples, the subject also incorporates a gastrointestinal disease or symptom, such as gastrointestinal dysfunction, e.g., diarrhea, constipation, nausea, abdominal distension, abdominal pain, vomiting, and the like. In some examples, the gastrointestinal disease or condition is due to hyperglycemia or diabetes.

A method of increasing the efficacy of a hypoglycemic agent comprising administering lysozyme and dossmate in combination with the hypoglycemic agent.

A method of reducing adverse effects of a hypoglycemic agent comprising administering lysozyme and dossmate in combination with the hypoglycemic agent.

In some examples of methods of increasing the efficacy of a hypoglycemic agent and methods of reducing adverse effects of a hypoglycemic agent, the hypoglycemic agent may be selected from insulin and analogues thereof, biguanide hypoglycemic agents, sulfonylurea hypoglycemic agents, thiazolidineone hypoglycemic agents, glinide hypoglycemic agents, alpha-glucosidase inhibitor hypoglycemic agents, glucagon-like peptide-1 (GLP-1) receptor agonist hypoglycemic agents, dipeptidylpeptidase-4 (DPP-4) inhibitor hypoglycemic agents, sodium-glucose transporter 2 (SGLT 2) inhibitor hypoglycemic agents, sodium-glucose transporter 1/2 (SGLT 1/2) dual inhibitor hypoglycemic agents, or other agents having hypoglycemic effects.

In some examples of methods of increasing the efficacy of a hypoglycemic agent and methods of reducing adverse effects of a hypoglycemic agent, lysozyme and dossmate are used before, after, or simultaneously with the use of the hypoglycemic agent.

In some examples of methods of reducing adverse effects of hypoglycemic agents, the adverse effects may be selected from gastrointestinal adverse effects, liver adverse effects, kidney adverse effects. Preferably adverse gastrointestinal reactions.

In some examples of the several methods described above, lysozyme and polyram may be combined in a variety of ways, including in separate formulations (combination package, co-administration), or in the presence of the same formulation (co-formulation). The specific formulation is not particularly limited, and may be various dosage forms suitable for pharmaceutical use, such as oral preparations, injections, inhalants, and the like.

In some examples of methods of increasing the therapeutic effect of a hypoglycemic agent and methods of reducing adverse effects of a hypoglycemic agent, the hypoglycemic agent may be combined with lysozyme and domamate in different forms, including in separate formulations (combination package, combination administration), or may be combined in the same formulation (compound formulation).

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

in some examples of the several methods described above, the daily amount of the driller's ester is 0.2 to 20g; preferably, the daily amount of the dossmate is 1-20 g; more preferably, the daily amount of the driller's ester is 1-10 g.

In some examples of the several methods described above, lysozyme is used in a daily amount of 0.2 to 20g; preferably, the daily dosage of lysozyme is 0.5-20 g; more preferably, the daily amount of lysozyme is 1 to 10g.

In some examples of the several methods described above, the trospium ester and lysozyme may be used 1-3 times per day, respectively.

The beneficial effects are that:

The inventor finds that lysozyme and the doesmaleate have remarkable synergistic effect in reducing blood sugar and can improve gastrointestinal dysfunction of patients with hyperglycemia. In addition, when lysozyme and the driller's horse ester are used together with other hypoglycemic drugs, the effect of the hypoglycemic drugs can be enhanced, and adverse reactions can be reduced. Thus, a combination of lysozyme and polysorbates is of particular clinical value for use in lowering blood glucose and hyperglycemia related diseases or conditions.

Detailed Description

Definition:

blood sugar: blood glucose generally refers to free glucose in blood plasma. Glucose in blood also binds to hemoglobin to form glycosylated hemoglobin, the proportion of which is generally proportional to the concentration of free glucose in plasma over a period of time and thus can also be used to evaluate blood glucose levels.

Hyperglycemia: normally the blood glucose concentration in the human body is kept constant in the range of 4-7mmol/L. The hyperglycemia is the hyperglycemia above the normal range.

Diabetes mellitus: diabetes is a metabolic disease, and persistent hyperglycemia is one of its fundamental features. Clinically, diabetes can be determined by standard medical diagnostic methods.

Lysozyme: lysozyme, i.e., lysozyme, is a lysozyme derived from animals, plants, microorganisms, or a recombinant of natural lysozyme. Examples of the recombinant human lysozyme include egg lysozyme, human lysozyme, recombinant human lysozyme and phage lysozyme. Lysozyme in the present invention also includes pharmaceutically acceptable salts thereof, such as hydrochloride, chloride, sulfate or amino acid salts and the like. Lysozyme has antibacterial and antiviral activities, and is recognized by the FDA as a safe substance (GRAS) in the united states.

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

Recently, lysozyme has been found to promote intestinal bacterial release of a Nod1 ligand that further promotes insulin transport efficiency by pancreatic beta cells (Zhang, 2019). This study provides a preliminary possibility for lysozyme treatment of hyperglycemia. However, the specific effects of lysozyme still need to be demonstrated. Through research, lysozyme has a certain effect, but the blood glucose reducing effect is still not ideal.

By combining the other drug, dosma ester, with lysozyme we have unexpectedly found that the hypoglycemic effect is significantly enhanced. Further researches also find that the combination has better application prospect, not only has the effect of reducing blood sugar, but also can be combined with the existing blood sugar-reducing medicines in the current market, and improves the effectiveness and safety of the existing medicines.

Polysmamate: the chemical name of the aluminum compound is Diominium heptabis (bisulfate) aluminum compound, and the molecular formula is Al ₇ (OH) ₁₄ (C ₂₈ H ₂₅ O ₃₆ S ₇ )[Al(OH) ₃ ] ₇ . Trospide is a gastric mucosa protectant that has been marketed for many years for the treatment of gastric and duodenal ulcers.

The invention is illustrated below by means of specific examples. Wherein both lysozyme and dossmate are available from North Hunan Weilman pharmaceutical Co.Ltd.

Example 1 preparation of lysozyme Polysimase Compound Capsule

50g of lysozyme stock is taken, 200g of lactose and 100g of microcrystalline cellulose are added, 5% polyvinylpyrrolidone ethanol solution is used as a binder, and the lysozyme particles are obtained after granulating and drying. Taking 500g of a doesyl maleate raw material, 100g of lactose, 50g of low-substituted hydroxypropyl cellulose and 2g of micro powder silica gel, granulating by using 5% polyvinylpyrrolidone ethanol solution as a binder, and drying to obtain doesyl maleate granules. Uniformly mixing lysozyme particles and the driller's ester particles, and encapsulating to obtain the lysozyme driller's ester composite capsule.

Example 2 preparation of lysozyme Polysimendate enteric composite particles

Taking 20g of lysozyme zymogen material, adding 50g of lactose, 50g of pregelatinized starch and 100g of microcrystalline cellulose, using 5% polyvinylpyrrolidone ethanol solution as a binder, granulating, coating an enteric coating, and drying to obtain lysozyme enteric particles. Taking 500g of a doesyl maleate raw material, 200g of lactose and 100g of low-substituted hydroxypropyl cellulose, using 5% polyvinylpyrrolidone ethanol solution as a binder, granulating, and drying to obtain doesyl maleate granules. And respectively taking the lysozyme enteric-coated particles and the dossmate particles, uniformly mixing, and bagging to obtain the lysozyme dossmate enteric-coated composite particles.

Example 3 Effect of combination of lysozyme and Polysmears on diabetic model animals

Animals: and selecting 10-week-old clean db/db transgenic mice with fasting blood glucose not lower than 18mmol/L, and weighing 40-50g. The feed is suitable for feeding for one week, and can be fed by free drinking water and standard feed.

Medicament: lysozyme (available from North Hunan Wilman pharmaceutical Co., ltd.), dossmate (available from North Hunan Wilman pharmaceutical Co., ltd.), sucralfate (commercially available).

Grouping: animals were randomly divided into model group, lysozyme group, doesyl ester group, sucralfate group, low-dose group of lysozyme doesyl ester, medium-dose group of lysozyme doesyl ester, high-dose group of lysozyme doesyl ester, low-dose group of lysozyme sucralfate, medium-dose group of lysozyme sucralfate, high-dose group of lysozyme sucralfate, 8 animals per group.

Administration: the model group was given 5% sodium methylcellulose by gavage. The corresponding drugs (mixed 5% methylcellulose solution) were administered by respective dosing in the case of a lysozyme group (60 mg/kg), a dossmate group (600 mg/kg), a sucralfate group (600 mg/kg), a lysozyme-dossmate low-dose group (60 mg/kg of lysozyme+6 mg/kg of dossmate), a lysozyme-dossmate medium-dose group (60 mg/kg of lysozyme+60 mg/kg of dossmate), a lysozyme-dossmate high-dose group (60 mg/kg of lysozyme+600 mg/kg of dossmate), a lysozyme-sucralfate low-dose group (60 mg/kg of lysozyme+6 mg/kg of sucralfate), a lysozyme-sucralfate medium-dose group (60 mg/kg of lysozyme+60 mg/kg of sucralfate), a lysozyme-sucralfate high-dose group (60 mg/kg of lysozyme+600 mg/kg of sucralfate). The administration was carried out 1 time a day, and the administration period was 4 weeks.

And (3) detection: after the completion of the administration, the animals were fasted for 8 hours, and the fasting blood glucose values of the animals of each group were measured by taking blood from the tail tip.

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

Table 1 fasting blood glucose measurement for animals of each group

Group of	Number of animals	Fasting blood glucose (mmol/L)
Model group	8	24.33±2.80
Lysozyme group	8	22.20±4.29
Polysimamate group	8	23.65±3.84
Sucralfate group	8	23.36±3.01
Lysozyme dossier ester low dose group	8	20.11±2.57*
Medium dose group of lysozyme tosomal ester	8	19.28±2.05**

Lysozyme sauceMa Zhigao dose group	8	17.84±2.65**
Lysozyme thiosugar aluminium low dose group	8	22.11±2.08
Medium dosage group of lysozyme sucralfate	8	22.71±1.44
High dose group of lysozyme sucralfate	8	21.31±1.67

Note that: p < 0.05 (×p) and p < 0.01 (×p) compared to model group.

Conclusion: the db/db mice are characterized by hyperphagia, obesity, hyperglycemia, insulin resistance and the like due to leptin gene deficiency, and are common spontaneous diabetes animal models. The experiment researches the short-term hypoglycemic effect of lysozyme, dossmate and sucralfate and the combination of several medicines, and discovers that the lysozyme can only slightly reduce blood sugar, but has no obvious difference compared with a model group, the hypoglycemic effect can be greatly enhanced after the combination of the lysozyme and the dossmate, and has very obvious difference compared with the model group, and the auxiliary hypoglycemic effect of the sucralfate is almost negligible.

EXAMPLE 4 Long-term effects of lysozyme and Polysimethide in combination with other hypoglycemic agents on diabetic model animals

Animals: selecting 10-week-old clean db/db transgenic mice with fasting blood glucose not lower than 18mmol/L, and weighing 40-50g, and 10-week-old wild C57BL/6J mice with the same nest, and weighing 20-25g. Animals were fed with the standard feed and with free water for one week.

Medicament: lysozyme particles (example 1), lysozyme enteric particles (example 2), domamate particles (example 1), acarbose tablets (commercially available), metformin hydrochloride tablets (commercially available).

Grouping: c57BL/6J mice were set as the normal group. db/db transgenic mice were randomly divided into model groups, metformin groups, acarbose groups, lysozyme dossmate metformin groups, lysozyme dossmate Sima Zhia carbobose groups, and lysozyme dossmate enteric groups. Each group was 10 animals.

Administration: normal and model groups were given normal saline by lavage. The corresponding drugs were administrated by gavage in doses (calculated as active ingredient) respectively, consisting of metformin (250 mg/kg), acarbose (50 mg/kg), lysozyme dossmate (100 mg/kg+300 mg/kg of dossmate), lysozyme dossmate metformin (100 mg/kg+300 mg/kg of dossmate+250 mg/kg of metformin), lysozyme dobis Sima Zhia carbobose (100 mg/kg+300 mg/kg of dossmate+50 mg/kg of acarbose), and lysozyme dossmate enteric (100 mg/kg+300 mg/kg of dossmate), wherein the lysozyme dossmate enteric group uses lysozyme enteric particles and the other groups containing lysozyme use lysozyme particles. The administration was carried out 1 time per day for 10 weeks.

Blood glucose determination: the fasting blood glucose values of each group of animals were determined at week 5 after the administration and after the completion of the administration (week 10), respectively.

Gastric emptying rate measurement: after blood sugar measurement, each group of animals fasted for 6 hours, was perfused with 2ml of 1.5mmol/L phenol red solution, and after anesthesia, the stomach was removed after laparotomy, pylorus and cardia were ligated, the stomach was cut along the greater curvature of the stomach, the stomach was flushed with distilled water, the stomach content was fixed to 20ml, and 20ml of 0.5mol/L sodium hydroxide solution was added, mixed well, and left to stand. 5ml of the supernatant was taken, and 0.5ml of 20% trichloroacetic acid was added thereto, followed by centrifugation to obtain a measurement solution. The absorbance of the measurement solution at a wavelength of 560nm was measured by an enzyme-labeled instrument, and the gastric emptying rate was calculated. Gastric emptying = (1-measured liquid absorbance/standard phenol red solution absorbance) ×100%.

Small intestine push rate measurement: the small intestine was taken out, laid on paper, and the distance from the pylorus to the ileum (total length of small intestine) and the distance from the pylorus to the farthest position from the phenol red (phenol red moving distance) were measured, and the small intestine thrust rate was calculated. Small intestine propulsion = (phenol red shift distance/small intestine full length) ×100%.

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

Table 2 fasting blood glucose determination for each group of animals

Note that: p < 0.05 (×p) and p < 0.01 (×p) compared to model group.

TABLE 3 gastric emptying rate and small intestine propulsion rate of animals of each group

Note that: p < 0.05 (×p) and p < 0.01 (×p) compared to model group.

Conclusion: the experiment researches the long-term hypoglycemic effect of the lysozyme-dossmate combination on the diabetic model mice, compares the long-term hypoglycemic effect with positive medicines of metformin and acarbose, and also performs three-medicine combination research. The combination of the lysozyme and the driller has long-term blood sugar reducing effect, and the combination of the lysozyme and the driller can also enhance the blood sugar reducing effect of the metformin or the acarbose, so that the enteric preparation has better effect. Metformin is a typical guanidine hypoglycemic agent, acarbose is a typical alpha-glucosidase inhibitor, which is a very commonly used hypoglycemic agent in clinic. The combination of the invention shows synergistic potential on the existing hypoglycemic drugs.

The present experiment also investigated the effect of several drugs on gastrointestinal function. It was found that db/db mice developed severe digestive dysfunction (e.g., decreased gastric emptying rate and decreased small intestine propulsion rate) around 20 weeks, but existing hypoglycemic drugs such as metformin or acarbose only worsened digestive function. Unexpectedly, however, the combination of the lysozyme driller's ester of the present invention can significantly increase the gastric emptying rate and the small intestine propulsion rate, i.e. significantly improve the digestive dysfunction of the diabetic model animal, and especially the enteric-coated preparation has better effect. In view of the wide variety of gastrointestinal adverse reactions such as digestive dysfunction, such as diarrhea, constipation, nausea, abdominal distension, abdominal pain, vomiting, etc. existing hypoglycemic agents at present, 20% of diabetics who clinically use metformin are reported to stop taking drugs because of the gastrointestinal adverse reactions, and therefore, the combination of the present invention has higher clinical value in improving compliance and tolerance of diabetics.

The present invention has been described in detail hereinabove, and variations or modifications thereof which would occur to persons skilled in the art are deemed to be within the spirit of the invention.

Claims

The use of a combination comprising lysozyme and dossmate for the manufacture of a medicament for lowering blood glucose.
Use of a combination comprising lysozyme and dossmate for the manufacture of a medicament for the prevention or treatment of a disease or condition associated with hyperglycemia.
The use according to claim 2, wherein the disease or condition associated with hyperglycemia is selected from one or more of diabetes, diabetic nephropathy, diabetic lower limb vascular disease, diabetic foot disease, diabetic retinopathy, diabetic macular degeneration, diabetic neuropathy or cardiovascular disease caused by hyperglycemia.
The application of the combination of lysozyme and dossmate in preparing the medicine for enhancing the drug effect of the hypoglycemic medicine and/or reducing the adverse reaction of the hypoglycemic medicine.
The use according to claim 4, wherein the hypoglycemic agent is selected from one or more of insulin and analogues thereof, biguanide hypoglycemic agents, sulphonylurea hypoglycemic agents, thiazolidineone hypoglycemic agents, glinide hypoglycemic agents, alpha-glucosidase inhibitor hypoglycemic agents, glucagon-like peptide-1 receptor agonist hypoglycemic agents, dipeptidyl peptidase-4 inhibitor hypoglycemic agents, sodium-glucose transporter 2 inhibitor hypoglycemic agents, or sodium-glucose transporter 1/2 dual inhibitor hypoglycemic agents.
The use according to claim 4, wherein the adverse effect is selected from gastrointestinal adverse effects, liver adverse effects, or kidney adverse effects.
A pharmaceutical composition comprises lysozyme and dossmate.
The pharmaceutical composition according to claim 7, wherein the weight ratio of lysozyme to dosmate in the pharmaceutical composition is 1:100 to 100:1.
the pharmaceutical composition according to claim 8, wherein the weight ratio of lysozyme to dosmate in the pharmaceutical composition is 1:20 to 20:1.
the pharmaceutical composition according to claim 8, wherein the weight ratio of lysozyme to dosmate in the pharmaceutical composition is 1:10 to 10:1.
the pharmaceutical composition of claim 7, further comprising an additional hypoglycemic agent.
The pharmaceutical composition of claim 11, wherein the additional hypoglycemic agent is selected from one or more of insulin and analogues thereof, biguanide hypoglycemic agents, sulfonylurea hypoglycemic agents, thiazolidineone hypoglycemic agents, glinide hypoglycemic agents, alpha-glucosidase inhibitor hypoglycemic agents, glucagon-like peptide-1 receptor agonist hypoglycemic agents, dipeptidyl peptidase-4 inhibitor hypoglycemic agents, sodium-glucose transporter 2 inhibitor hypoglycemic agents, or sodium-glucose transporter 1/2 dual inhibitor hypoglycemic agents.
The pharmaceutical composition according to claim 7, further comprising pharmaceutically acceptable excipients.
The pharmaceutical composition of claim 7, which is an oral formulation, an injectable formulation, or an inhaled formulation.
The pharmaceutical composition of claim 7, which is a solid formulation, a semi-solid formulation, or a liquid formulation.
The pharmaceutical composition of claim 7, which is a constant release formulation, a delayed release formulation, a sustained release formulation, or a controlled release formulation.
The pharmaceutical composition of claim 7, wherein the release forms of lysozyme and dossmate may be the same or different.
The pharmaceutical composition of claim 17, wherein the dosma ester is a gastric-soluble release.
The pharmaceutical composition of claim 17, wherein the lysozyme is enteric released.
A combination comprising lysozyme and dossmate.
The combination according to claim 20, wherein the lysozyme is in the form of a separate formulation and the domamate is in the form of a separate formulation.
The combination according to claim 21, wherein the lysozyme and/or the dossmate is an oral, injectable or inhaled formulation.
The combination according to claim 21, wherein the lysozyme and/or the dossmate is a solid, semi-solid or liquid formulation.
The combination according to claim 21, wherein the lysozyme and/or dosma ester is a constant release formulation, a delayed release formulation, a sustained release formulation or a controlled release formulation.
The combination according to claim 20, wherein the weight ratio of lysozyme to dosmate in the combination is from 1:100 to 100:1.
The combination according to claim 20, further comprising an additional hypoglycemic agent.
The combination according to claim 26, wherein the additional hypoglycemic agent is selected from one or more of insulin and analogues thereof, biguanide hypoglycemic agents, sulphonylurea hypoglycemic agents, thiazolidineone hypoglycemic agents, glinide hypoglycemic agents, alpha-glucosidase inhibitor hypoglycemic agents, glucagon-like peptide-1 receptor agonist hypoglycemic agents, dipeptidyl peptidase-4 inhibitor hypoglycemic agents, sodium-glucose transporter 2 inhibitor hypoglycemic agents, or sodium-glucose transporter 1/2 dual inhibitor hypoglycemic agents.
A method of lowering blood glucose comprising administering to a subject in need thereof effective amounts of lysozyme and dossmate.
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 dossmate.
The method of claim 29, wherein the disease or condition associated with hyperglycemia is selected from the group consisting of diabetes, diabetic nephropathy, diabetic lower limb vascular disease, diabetic foot disease, diabetic retinopathy, diabetic macular degeneration, diabetic neuropathy, and hyperglycemia-induced cardiovascular disease.
A method for enhancing the therapeutic effect of hypoglycemic agents comprises the step of combining lysozyme and dosma ester with hypoglycemic agents.
A method of reducing adverse effects of a hypoglycemic agent comprising administering lysozyme and dossmate in combination with the hypoglycemic agent.
The method of claim 32, wherein the adverse reaction is selected from one or more of gastrointestinal adverse reaction, liver adverse reaction, kidney adverse reaction.
The method of any one of claims 28-33, wherein the lysozyme and the dossmate are used in a weight ratio of 1:100 to 100:1.
The method of any one of claims 28-33, wherein the daily amount of the driller's ester is 0.2-20 g.
The method of any one of claims 28-33, wherein the lysozyme is used in an amount of 0.2 to 20g per day.