CN110302359A - A kind of preparation reducing blood glucose, preparation method and applications - Google Patents

Abstract

The invention discloses the preparation that one kind can reduce blood glucose, active constituent, which contains, extracts resulting polypeptide by raw material of pluck.The preparation provided by the invention for reducing blood glucose can be effectively reduced the blood glucose of hyperglycemic patients, can be used cooperatively individually or with existing hypoglycemic drug, it is possible to solve the problems, such as the insulin resistance that patients with NIDDM occurs.

Description

A kind of preparation reducing blood glucose, preparation method and applications

Technical field

The present invention can reduce the preparation of blood glucose, also relate to the preparation method and application of said preparation.

Background technique

Diabetes are the diseases of the metabolic disorder characterized by hyperglycemia, pathogenesis be due to insulin secretion not Caused by foot or insulin action obstacle.Diabetes system is after cardiovascular disease and tumour, it has also become world's human life and health In the presence of three big high risk factors of threat.According to epidemiologic data, the whole world shares 4.15 hundred million people and suffers from diabetes at present, it is contemplated that arrives The year two thousand forty is up to 6.42 hundred million.About 1.5 hundred million, of China diabetic accounts for about population 11.8%, occupies the whole world first.In addition, population of China Simple blood glucose increased perosn (prediabetes) accounts for about adult population 30%, it is seen that the preventing and controlling for diabetes are a Xiang Jiqi Important task.

Diabetes can be divided into I type and type II diabetes, and Type I diabetes is also known as insulin dependent diabetes, since body produces There is beta Cell of islet damage in raw immune response, and insulin secretion reduces or lacks, and common children and teenager account for about diabetes Patient 5%.Type II diabetes is also known as non-insulin-dependent diabetes mellitus, and early insulin secretion is normal or slightly reduces, and high blood is presented Sugar, predominantly presentation dysglycemia, the state of an illness are generally slowly in progress, and patient's islet cells is reduced with annual 5% speed, through 10 After~12 years, about 50% islet beta cell function can be lost.Type II diabetes with adult common, account for about diabetic 90~ 95%.

It is often spy to insulin resistance to generate other than being related to islet beta cell function obstacle in type II diabetes Sign.So-called insulin resistance refers to that the target organ of insulin action has dropped the susceptible of insulin, i.e. normal dose Insulin generate be lower than natural biological effect state.Visible insulin dosage is increasing in clinic, but Blood sugar recovery normal level is not can control still.

Repulsion or dysbolism of the diabetic due to body to the various physiological actions of insulin.Normal insulin point Blood is secreted, the effects of hypoglycemic or tissue is using sugar cannot be generated, each histoorgan is caused to be involved.The complication of diabetes has Factors, including heredity, age, gender, different blood glucose levels etc..Diabetes find retinal complication 31.5%, kidney Disease 39.7%, neuropathy 51.1%, coronary heart disease 25.1%, hypertension 51.8%.

Treating diabetes generally use diabetes education, Blood sugar self-measure, diet control, appropriate exercise and drug therapy Etc. the integration scenario of so-called " five drive carriage ".

Diabetes drug treatment:

The treatment of diabetes since nineteen twenty-one finds insulin, has had short at present still using insulin as main method Effect, middle effect, long-acting various insulin preparations, curative effect are also affirmed very much, but to so far, insulin is still needed to using injection side Formula, there has been no breakthroughs for oral preparation, since existing insulin preparation is exogenous, it is necessary to it uses throughout one's life, additionally due to Diabetic often generates insulin-resistant states, is that current insulin therapy has obvious shortcoming.

Other treatment diabetes medicaments:

1. reducing sugared source

(1) melbine: inhibit intestinal wall cell to glucose absorption and gluconeogenesis, tissue glycogen is promoted to decompose.

(2) alpha-glucosidase inhibitor (Glucobay etc.): can inhibit glucosidase activity, slows down glucose generation, drops Hypoglycemia.

2. promoting insulin secretion

(1) sulfonylureas (such as this urea of Gree): promote beta Cell of islet (not completely disappearing function) excreting insulin.

(2) incretin.

3. insulin sensitizer (Rosiglitazone), can promote blood glucose to absorb, accelerate Glycogen synthesis, enhances insulin sensitivity Property.

Summary of the invention

It is an object of the present invention to provide the preparations that one kind can reduce blood glucose.

An object of the present invention is achieved through the following technical solutions: one kind can reduce the preparation of blood glucose, activity at Divide containing using pluck as the resulting polypeptide of raw material extraction.

The pluck refers to one or both of internal organ of mammal, including liver, spleen and pancreas Combination.

The molecular weight of the polypeptide is within 10000Da.Preferably, the molecular weight of the polypeptide 1000-8500Da it Between.

Content of peptides is 5-20mg/ml, preferably 6-10mg/ml in the preparation.

The preparation is oral preparation.It further include available auxiliary material on pharmacy or food in oral preparation, such as water, seasoning Agent etc..Specifically, the oral preparation can be the conventional formulation of pharmacy and food, including freeze dried powder, oral solution, capsule Agent, tablet etc..

The preparation is injection, can be liquid injection agent, injectable powder, injection tablet etc..

The second purpose of the present invention is to provide the preparation methods of the above-mentioned preparation for reducing blood glucose.

Specifically, described the preparation method is as follows: take pluck after cryogenic pulverization, membrane filtration to get.

When the preparation is injection, need to be purified under conditions of meeting injection production requirement.Similar side Method can refer to the purifying of hepatocyte growth promoting factor.

The three of the object of the invention are to provide the application of above-mentioned preparation.Specifically, the above-mentioned preparation conduct for reducing blood glucose The application of the hypoglycemic complementary food of hyperglycemic patients or drug.

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

1. the preparation provided by the invention for reducing blood glucose can be effectively reduced the blood glucose of hyperglycemic patients, can individually or with Existing hypoglycemic drug is used cooperatively.

2. preparation provided by the invention can be used for human I-type and type II diabetes treatment, insulin or other tools can be cooperated There is the comprehensive drug treatment of hypoglycemic effect.Experiments have shown that having to Experimental diabetic model rat reduces blood glucose and promotion pancreas The effect that island element generates.Preliminay clinical trials show preparation category natural prodcuts provided by the invention, without any side effects, use Safety.Preliminay clinical trials are shown, using hypoglycemic medicine or combine insulinization through many years to II type diabetic, but curative effect It is unsatisfied with and insulin resistance may have occurred, by adding with after preparation provided by the invention, rough estimates was controlled through 6~12 months It treats, it is raised efficient up to 70% or so to fasting blood-glucose is reduced.Especially to insulin resistance person, pancreas islet can be gradually decreased Plain dosage, or even insulin is finally deactivated, it still can control fasting blood-glucose and reach ideal value.It can be seen that the present invention is for treatment Diabetes have extensive prospect and potentiality, and oral effective, without any side effects.As long as adhering to medication 6 months or more, have May solve the problems, such as the insulin resistance that patients with NIDDM occurs.

Specific embodiment

Embodiment one

Fresh animal internal organ (liver of the mammals such as pig, ox, sheep, spleen, pancreas) is taken, fritter is first cut into, is subsequently placed in Rubber mill is homogenized about 6 minutes, is placed on cryogenic freezing 3 days, using ultracentrifugation, ultrafiltration, chromatography, as animal Internal organ extract (abbreviation oral solution), every milliliter of 5mg containing peptide material.

Embodiment two

Fresh animal internal organ (liver of the mammals such as pig, ox, sheep, spleen, pancreas) is taken, fritter is first cut into, is subsequently placed in Rubber mill is homogenized about 8 minutes, is placed on cryogenic freezing 5 days, using ultracentrifugation, ultrafiltration, chromatography, as animal Internal organ extract (abbreviation oral solution), every milliliter of 15mg containing peptide material.

Embodiment three

Fresh animal internal organ (liver of the mammals such as pig, ox, sheep, spleen, pancreas) is taken, fritter is first cut into, is subsequently placed in Rubber mill is homogenized about 15 minutes, is placed on cryogenic freezing 4 days, as dynamic using ultracentrifugation, ultrafiltration, chromatography Object internal organ extract (abbreviation oral solution), every milliliter of 20mg containing peptide material.

Example IV

In embodiment one, two and three, gained pluck extract is purified using conventional process for preparation of injection And it is prepared into liquid injection agent, injectable powder, injection tablet.

The therapeutic effect of one oral solution streptozotocin inducing experimental diabetes rat model of test example

The experimental rat diabetes rat model induced using streptozotocin (STZ), and with pluck extract (abbreviation oral solution) observes its influence to fasting blood-glucose and generation insulin is reduced.

Materials and methods

Animal selection: health pure lines SD rat male 30, it is about 180~220 grams of weight, 18~25 DEG C of room temperature, relatively wet It is fed under the conditions of degree 50%~80%, freely ingests, drinks water.

Experimental method: 30 SD rats are randomly divided into 3 groups, every group each 10, wherein experimental comparison group and treatment group are each 0.1mol/L citric acid-sodium citrate buffer solution (pH4.4) is dissolved in using STZ, is made into 1% solution, by 60mg/kg single abdominal cavity Drug administration by injection, blank group then apply the sterile citric acid-sodium citrate buffer solution intraperitoneal injection of the 0.1mol/L of relative volume.Experiment Treatment group rat since intraperitoneal injection STZ3 days after feeding one oral solution of embodiment, 2 times a day, each 1.0ml, totally 45 days, And control group is then fed with same volume physiological saline 1.0ml, blank group does not give injection STZ and feeds liquid.It is before injection and oral Tail vein is cut after feeding once a week and takes blood, totally 6 weeks, detects blood glucose, each group rat detects periphery blood insulin water after 6 weeks It is flat.

Experimental result

The comparison that insulin generates after the pretherapy and post-treatment change of blood sugar of 1 groups of animals of table and treatment

※ treatment group is after treatment 45 days, and average blood sugar is remarkably decreased before relatively treating, P < 0.05.And insulin level is It is restored to close to blank group level.Treatment group's insulin restores compared with control group significant difference, P < 0.05.

From the above results, it can be seen that oral solution is after treatment in 45 days, treatment group's fasting blood-glucose is gradually recovered, and insulin secretion Increased, oral solution is prompted to have promotion repair to the damage of islet cells.

The influence that two pluck extract of test example controls patients with NIDDM fasting blood-glucose

Clinical basic document

1. diabetic's selected condition

Patients with NIDDM meets type II diabetes metabolism of blood glucose according to " version Guidelines for Management of Diabetes Mellitus in 2017 " patient Abnormal three norms, i.e. fasting blood-glucose > 7.0mmol/L, 2h-plasma glucose > 11.0mmol/L, glycosylated hemoglobin > 7.0% (being vein method), before being selected in addition to having hypertension, Hyperlipidemia, without serious diabetic complication, such as glycosuria kidney Insufficiency, retinopathy, digital ulcer etc..

2. group case basic condition

28 are patients with NIDDM, and male 12, women 16, course of disease 3-24 is differed, and have been grown before selected Phase takes hypoglycemic agent (melbine, Gree this urea etc.) but fasting blood-glucose still fluctuates between 7.2~14.0mmol/L, accounts for 27 (96.2%), 2h-plasma glucose > 11.0mmol/L person 25 (89.3%), glycosylated hemoglobin > 7.0%23 (82.1%).In this group 28, before being selected in addition to oral hypoglycemic, and combine subcutaneous injection of insulin (each 8-20 Unit, 2 times a day) person 6 (21.4%).

3. therapeutic scheme

According to the situation for controlling fasting blood-glucose before patient selection, add with one oral solution of embodiment, using 3 kinds of oral solution dosage Method: slight person 3 times a day, each 10ml；The light to moderate each 20ml of person, 2 times a day or takes 20ml morning, after dinner Take 30ml；The state of an illness is relatively long, and the person that shares subcutaneous injection of insulin, each 30ml, 2 times a day.The medication course for the treatment of: every 3 months It is as a treatment course, generally take 2~4 courses for the treatment of.Monthly once, blood capillary can be used in remaining time for blood count (vein method) during treatment The observation of tube method self detecting blood sugar.Glycosylated hemoglobin, every 3 months primary.

4. curative effect judges

According to the actual conditions of this research, the curative effect in terms of controlling blood glucose to oral liquid treatment type II diabetes judges, point It is as follows for three kinds of situations.

4.1 is effective:

Patients with NIDDM, after making a definite diagnosis, applied originally hypoglycemic drug (such as melbine) more than one, but Fasting blood-glucose still fails to fully control, and fluctuates in 7.0~11.0m mol/L, after being added one oral solution of embodiment, fasting blood-glucose It can stablize in 4.4~7.0m mol/L continuous 3 months or more persons, can be described as effective.

Another situation is that selected case removes Long-term Oral hypoglycemic agent beyond the region of objective existence, subcutaneous injection of insulin has been shared, every time 8 ~20 units, daily 2~3 times, but fail to fully control blood glucose and reach ideal value < 7.0m mol/L, added embodiment flatly Liquid June~12 month are taken, can control fasting blood-glucose fluctuation, and gradually decrease subcutaneous injection of insulin dosage or even deactivated pancreas Island element only retains after taking one oral solution of antidiabetic drug and embodiment, also can determine whether to be effective.

4.2 effectively:

Above-mentioned two classes patient only oral hypoglycaemic drug or has combined the patient for answering subcutaneous injection of insulin, through adding reality Apply one oral solution June of example~after December, fasting blood-glucose can reach ideal value substantially, but have 7.0~8.5mmol/L of slight fluctuation Between, or preceding decline is relatively treated up to 50% blood glucose person, it can be described as effectively.

4.3 is invalid:

The above two original patient using antidiabetic drug, it is not up to above-mentioned effective or have when adding oral solution 6 months or more Ideal value person is imitated, referred to as in vain.Selected case should be rejected by adding one oral solution of embodiment and being not up to 6 months automatic drug withdrawal persons, Not count.

4.4 treatment results

4.4.1 curative effect: this group of case, be from April, 2018~2019 year June, added oral liquid treatment reach June~ 12 months persons, clinical efficacy see the table below

2 oral solution of table controls blood sugar influence to type II diabetes

4.4.2 side effect: oral solution is good in taste, and patient can receive, and has no any side effect.

4.4.3 typical case

XX is opened, it is male, 62 years old.Find that fasting blood-glucose increases (13.8m mol/L) in July, 2007, thereafter oral administration of metformin And Gree this urea, but fasting blood-glucose still fluctuates 11.0~13.5m mol/L, 13.0~16.5mmol/L of 2h-plasma glucose to 2017 May in year adds is infused with insulin skin, and daily 16 unit is injected in two times, but fasting blood-glucose still has fluctuation not can control.In 2018 4 The moon starts to take one oral solution of embodiment on the 11st, and 3 times a day initially, each 10ml is changed to each 20ml after 2 months, sooner or later each Once.Thereafter fasting blood-glucose is gradually controlled in range of normal value.Start to deactivate in March, 2019 (after taking oral solution 11 months) Insulin skin note, is changed to increase oral Ge Qite, and daily 2.To on July 18th, 2019, insulin has been deactivated 4 months, on an empty stomach Blood glucose still controls between 6.8~7.8mmol/L.Observation treatment is continued at present.

Xiao XX, male, 70 years old.It makes a definite diagnosis type II diabetes within 2000, has taken various hypoglycemic drugs for many years, fasting blood-glucose, Postprandial blood sugar is unstable, starts within 2012 to add with insulin 2 times a day, every time 12~20 unit, but fasting blood-glucose does not reach yet It to ideal value, fluctuates between 14~16m mol/L, adds one oral solution of embodiment since in the July, 2018, each 30ml, often Day 2 times, fasting blood-glucose is gradually stable normal thereafter, until having reduced application dose at the beginning of 7 months 2019, is infused by the daily skin of insulin 40 units are kept to daily 30 unit, and nearly 3 wheat harvesting period comes, and fasting blood-glucose controls between 7.5~8.5 units.

Deng XX, male, 43 years old.It makes a definite diagnosis type II diabetes in June, 2015, starts in July, 2015 to take melbine over 3 years on an empty stomach Blood glucose still fluctuates between 9.5~11.7m mol/L, and in September, 2018 starts to add one oral solution of embodiment, 2 times a day, every time 20ml.Thereafter fasting blood-glucose is gradually stable, until in May, 2019 (i.e. plus with oral solution 8 months after), fasting blood-glucose complete stability exists 7.0m mol/L is hereinafter, 2h-plasma glucose 11.5m mol/L glycosylated hemoglobin 6.3%.Observation treatment is continued at present.

Three normal person of test example and type II diabetes people the peripheral blood insulin-like growth factor before and after oral liquid treatment (IGF-1) comparison and its meaning detected

(1) purpose:

In order to inquire into the possibility mechanism of action that oral solution treats diabetic's regulating and controlling blood sugar.We are to 10 normal person's (nothings Diabetic history) and 10 patients with NIDDM IGF-1 in peripheral blood before application oral liquid treatment and after treatment 6 months change Amount is studied.

(2) material and method:

Reagent: human insulin-like growth factor (IGF-1) kit, R&D company, the U.S..

Operating procedure: separation serum or blood plasma are pre-processed before detection, IGF-1 are allowed to release from binding protein； Test buffer RD1-53 adds 150 holes μ l/；Each hole 50 μ l/ of measuring samples, standard items is taken to be added in ELISA Plate respectively, 2~8 DEG C It places 2 hours；With cleaning solution board-washing 4 times；The people's IGF-1 binding antibody for adding 200 holes μ l/ cold is added in ELISA Plate, and 2~8 DEG C put It sets 1 hour；With cleaning solution board-washing 4 times；After thoroughly blotting ELISA Plate, every hole adds 200 hole μ l/ of color developing agent, is stored at room temperature 30 minutes； Every hole adds 50 μ l of terminate liquid；50nm/570nm surveys A value.

As a result it calculates: using microplate reader curve quant program, according to the concentration of standard items and corresponding test sample A value, directly Reading the concentration of product to be tested and multiplying corresponding extension rate is test sample ultimate density.

(3) experimental result:

1. whether containing IGF-1 in oral solution.

IGF-1 content in 3 oral solution of table

IGF-1 content in the oral liquid formulations provided by 3 embodiments of detection, as a result as shown in Table 3, oral liquid formulations In fail to detect IGF-1.

2. the peripheral blood IGF-1 detection before and after oral liquid treatment of normal person and patients with NIDDM (each 10) is compared

4 normal person of table and type II diabetes human peripheral IGF-1 compare

※ normal person is compared with diabetes patient (before treatment) peripheral blood IGF-1 level, P < 0.05；Diabetes patient applies mouth It takes before liquid is treated and compares after treatment 6 months, P < 0.05.

Shown in 4 result of table as above, before 10 one oral liquid treatments of type II diabetes people Application Example and after treatment 6 months Compare, IGF-1 is significantly increased.IGF-1 has a kind of important feature and insulin homolog.IGF-1 and pancreas islet are known as different receptors, but There is 60% amino acid sequence homologous.The receptor of IGF-1 is 1000 times to the affinity of IGF-1, much larger than insulin receptor to pancreas Island element affinity is 100 times.Insulin mainly adjusts glycometabolism, and IGF-1 then plays the part of horizontal stable in maintenance glycometabolism body And balance, IGF-1 can promote intake of the peripheral tissues to sugar, IGF-1 can also inhibit the generation (decomposition) of hepatic glycogen.IGF-1 energy It is sensitive to glycometabolism to increase insulin, in short, IGF-1 and insulin both participate in glycometabolism, it is to have to the disorder for correcting glycometabolism Complementary effect.Shown in result from table 3, pluck extract has not been found due to passing through repeated detection The ingredient of IGF-1.But animal experiments show that having reduces blood glucose and promotion insulin generation effect.Further clinical test confirms Blood glucose can not only be reduced, and after taking 6~12 months, peripheral blood IGF-1 significantly increases in reception test person's body, P < 0.05.As it can be seen that the characteristics of pluck extract of the invention may be containing insulin-like growth factor-i be promoted.And this Invention can take orally, without any side effects.It is easy to use, it is possible to the new method as treatment type II diabetes.

Claims (10)

1. the preparation that one kind can reduce blood glucose, characterized in that its active constituent includes resulting using pluck as raw material extraction Polypeptide.

2. the preparation according to claim 1 for reducing blood glucose, characterized in that the pluck refers to that mammality is dynamic The internal organ of object.

3. the preparation according to claim 2 for reducing blood glucose, characterized in that the internal organ include liver, spleen and pancreas One or both of dirty combination.

4. the preparation according to claim 1-3 for reducing blood glucose, characterized in that the molecular weight of the polypeptide exists Within 10000Da.

5. the preparation according to claim 4 for reducing blood glucose, characterized in that the molecular weight of the polypeptide is in 1000- Between 8500Da.

6. the preparation according to claim 1-5 for reducing blood glucose, characterized in that content of peptides in the preparation For 5-20mg/ml.

7. the preparation according to claim 6 for reducing blood glucose, characterized in that content of peptides is 6- in the preparation 10mg/ml。

8. the preparation according to claim 5 for reducing blood glucose, characterized in that the preparation is oral preparation or injection Agent.

9. the preparation method that one kind can reduce the preparation of blood glucose, characterized in that take pluck after cryogenic pulverization, filter membrane mistake Filter to get.

10. the described in any item preparations for reducing blood glucose of claim 1-8 are as the hypoglycemic complementary food of hyperglycemic patients Or the application of drug.