CN112274524A

CN112274524A - Application of panaxadiol saponin Rb component in preparing medicine for preventing and treating metabolic disorder related diseases

Info

Publication number: CN112274524A
Application number: CN202010975465.4A
Authority: CN
Inventors: 连晓媛; 张治针; 池彬彬; 朱荣遥; 钱恒
Original assignee: Zhejiang University ZJU
Current assignee: Zhejiang University ZJU
Priority date: 2017-08-24
Filing date: 2017-08-24
Publication date: 2021-01-29
Also published as: CN107441104B; CN107441104A

Abstract

The invention provides an application of a panaxadiol saponin Rb component in preparing a medicament for preventing and treating metabolic disorder-related diseases. The medicine is prepared with panaxadiol saponin Rb component as single active component or together with other active compounds, wherein the Rb component mainly comprises five panaxadiol saponins (Rb)₁、Rb₂、Rb₃Rc and Rd) accounting for 50-98% of the Rb component, and other active compounds including various substances capable of reducing blood sugar level, traditional Chinese medicine active substances, natural products, artificially synthesized compounds and in-vivo active substances. The invention corrects the diabetes metabolic disorder by taking independent blood sugar level as an action mechanism, can enhance the mitochondrial function and the oxidation-reduction balance capability of a diabetic patient, and can prevent and treat diseases and pathological symptoms which take metabolic disorder, particularly mitochondrial dysfunction as a common pathological mechanism for mitochondrial damage and metabolic disorder related pathological symptoms caused by pathological factors other than diabetes. The medicine is safe and effective.

Description

Application of panaxadiol saponin Rb component in preparing medicine for preventing and treating metabolic disorder related diseases

The present case is application number: 201710735766.8, filing date: 2017.8.24, title of the invention: application of panaxadiol saponin Rb component in preparing medicine for preventing and treating diabetes complication and metabolic disorder related diseases is provided.

Technical Field

The invention belongs to the field of medicine, and relates to application of a panaxadiol saponin Rb component (hereinafter referred to as Rb component) in preparing a medicine for preventing and treating diabetes complications and metabolic disorder related diseases. More specifically, the Rb component is used for improving the metabolic symptoms of polydipsia, polyphagia and polyuria of a diabetic patient, preventing and treating the central and peripheral complications of diabetes and other diseases and symptoms related to metabolic disorders (including central neurodegenerative diseases and neurotoxicity caused by cancer treatment).

Background

Pre-diabetes and diabetes are well-recognized global epidemics, and diabetes and its complications have become the third leading killer threatening human life health after cardiovascular and cerebrovascular diseases, cancer. With the prevalence of global obesity and the aging of the world population, a metabolic storm is in mid-life (Climacteric.2017 Feb; 20(1):11-21). At present, allWithin the sphere are 3 hundred million 1600 million pre-diabetic patients and 3 hundred 8700 million diabetic patients, respectively, of which at least 2 million people have neuropathy (Neuron.2017Mar 22; 93(6):1296-1313). According to the International Diabetes union (IDF) survey, it was estimated that there are roughly 5 billion diabetic patients worldwide by 2025 years (International Diabetes Federation, 2014). At present, more than 9240 thousands of diabetes patients exist in China, and have become the first major country of diabetes in the world. More worrying about that the standard reaching rate of blood sugar of common people is less than 30 percent while the prevalence rate of diabetes in China is continuously increased. Diabetes mellitus is a group of metabolic diseases characterized by defects in insulin secretion or insulin action due to insulin resistance or both leading to hyperglycaemic and carbohydrate metabolism disorder syndromes accompanied by long-term damage and dysfunction or loss of different organs (in particular eyes, kidneys, nerves, heart, blood vessels) and associated diabetic complications (ii) ((iii))Curr Pharm Des.2015；21(34):4980-8.32；Diabetes Care.2017Jan；40(1):136-154.；World J Diabetes. 2015Oct 10; 6(13):1246-58.). The clinical manifestations of diabetic complications are complex and diverse, and involve a variety of diseases, common ones are: peripheral neuropathy, microvascular complications including Diabetic Nephropathy (DN) and retinopathy (DR), macrovascular complications such as coronary heart disease, cerebrovascular disease, peripheral vascular disease, etc. Diabetes is the leading cause of new blindness and greatly increases the risk of neuropathic pain, heart disease and renal failure. Geriatric diabetes also increases the risk of osteoporosis and cancer. In the process of diabetes occurrence and development, the incidence rate of diabetic complications is up to 96 percent, and the diabetic complications are the most known diseases at present; about 10 years after the diabetes mellitus happens, at least one complication can happen to 30-40% of patients, and once the complication is generated, the drug treatment is difficult to reverse. Therefore, the diabetic complications seriously reduce the survival quality of the patients, are also the main cause of death and disability of the diabetic patients, and greatly increase the risk of disability and death of the patients.

Diabetic peripheral neuropathy is one of the most common chronic complications of diabetes, a group of sensory and autonomic nervesPeripheral neuropathy which becomes the main clinical manifestation, like diabetic nephropathy and diabetic retinopathy, seriously affects the quality of life of patients: (Diabetes Care.2017 Jan; 40(1):136-154.). Sensory neuropathy is characterized by pain, paresthesia (burning discomfort) and loss of sensation: (Neuroendocrinology.2013；98(4):267-80；Diabetes Care.2017 Jan; 40(1) 136-. Patients lose normal daily activity due to pain and often experience sleep disturbances, anxiety and depression, which severely reduces the quality of life of the patient (Nat Rev Dis Primers.2017Feb 16; 3: 17002.). As diabetic peripheral neuropathy progresses, decreased or even loss of pain sensitivity and impaired motor balance and postural instability occur, which all contribute to diabetic foot, fall and fracture in patients: (Diabetes Care.2017 Jan; 40(1):136-154.). The overall prevalence of neuropathic pain symptoms in neuropathic patients is 21% (ii)Nat Rev Dis Primers. 2017 Feb 16；3:17002.). Wherein the prevalence rate of diabetic peripheral neuropathy in China is as high as 56.5%. Diabetic autonomic neuropathy is closely related to myocardial infarction, malignant arrhythmia, and sudden death (Neuroendocrinology.2013；98(4):267-80.；Diabetes Care,2010,33(2):434-441；Diabetes Care.2010Feb；33(2):434-41；World J Diabetes. 2015Oct 10; 6(13):1246-58.). Studies have shown that patients with type 2 diabetes are at a 2-4 times higher risk of cardiovascular disease and stroke than the general population, with a 5-10 year reduced lifespan. In addition, the main clinical manifestations of diabetic autonomic neuropathy are hypoglycemia, resting tachycardia, postural hypotension, gastroparesis, constipation, diarrhea, fecal incontinence, erectile dysfunction, neurogenic bladder dysfunction, and increased or decreased sweating, gastrointestinal tract, urogenital tract and perspiration dysfunction, all of which severely reduce the quality of life of the patient(s) ((Diabetes Care.2017 Jan; 40(1):136-154.). In type 2 diabetic patients, the prevalence of diabetic autonomic neuropathy also increases with the duration of diabetes, and may occur in 60% of patients after 15 years. In the United states and Europe, pre-diabetes and type 2 diabetes (T2DM) are the most common forms of peripheral neuropathyFor this reason, at least 50% of diabetic patients, including type 1 diabetic patients, develop some form of neuropathy during their lifetime (neuron.2017 Mar 22; 93(6): 1296-. At least 20% of type 1 diabetic patients with a history of more than 20 years develop distal polyneuropathy. Newly diagnosed type 2 Diabetes patients may have at least 10% -15% of distal polyneuropathy, and the incidence of disease course will increase to 50% over 10 years (Diabetes Care.2017 Jan; 40(1): 136-154.).

Recent studies have demonstrated that the central nervous system also does not survive the deleterious effects of diabetes. Diabetic encephalopathy characterized by specific cognitive and neurobehavioral disorders is gradually perceived as a decline in memory and resolution, sleep disorders, depressed mood, and the likeCurr Pharm Des.2015; 21(34):4980-8.32). In addition, the incidence of various neurodegenerative diseases (including Alzheimer's disease, vascular dementia, Parkinson's syndrome and Huntington's disease) in type II diabetic patients is significantly higher than that in non-diabetic age-matched populations (Lancet Neurol.2004 Mar; 3, (3) 169-78), in which the incidence of AD is increased by 2 times (J Clin Invest2013Feb 1; 123(2):531-539.). In particular, it is noted that more and more studies have demonstrated that metabolic disorders, in particular mitochondrial dysfunction and the oxidative stress injury and inflammatory response triggered thereby, are common pathological features of pain caused by diabetic peripheral neuropathy and peripheral neuropathic pain caused by chemotherapeutic drugs, and also common pathological mechanisms of central neurodegenerative diseases (including senile dementia, vascular dementia, Parkinson's disease, multiple sclerosis, etc.) (J Clin invest.2013Feb 1; 123(2): 531-539). Among them, senile dementia has a metabolic disorder phenotype in brain similar to that of type 2 diabetes, and thus senile dementia is called developing intracerebral diabetes, also called type 3 diabetes (Biochem Pharmacol.2014 Apr 15; 88(4): 548-59.; Eur Neuropychlorophharmacol.2014 Dec; 24(12): 1954-60.; Neurol Sci.2015 Oct; 36(10): 1763-9.). It is noted that neurological disorders (epilepsy, schizophrenia, depression, etc.) are also closely related to metabolic disorders (Mitochondrion.2012 Jan; 12(1): 35-40).Lancet Neurol.2015 Sep；14(9):956-66.Neuromolecular Med.2015Dec；17(4):404-22.)。

The prevention and treatment of diabetic complications and neurodegenerative diseases has long been an international problem. Despite decades of research, there are currently no other methods and drugs available globally for the treatment of diabetic peripheral neuropathy, in addition to improving lifestyle and controlling blood glucose in patients: (Diabetes Care.2017 Jan; 40(1):136-154.). Cochrane review clinical studies show that tight glycemic control can reduce the incidence of type I diabetic neuropathy, but not type II diabetic neuropathy, similar studies in which results are shown in the prevention of diabetic autonomic neuropathy: (Diabetes Care.2017 Jan；40(1):136-154；Neuron.2017Mar 22; 93(6):1296-1313). In the United states, all clinical trials aimed at altering diabetic neuropathy have failed and major pharmaceutical companies have now left the area of diabetic neuropathy ((R))Diabetes Care.2017 Jan; 40(1):136-154.). Therefore, for preventing and treating diabetic complications, the search for a medicament which takes independent blood sugar as an action mechanism is not slow.

The pathological mechanism is a lighthouse that directs the development of effective drugs. Neuropathy and vasculopathy are not only important complications of diabetes, but also closely related to neurodegenerative diseases, so that protection of nerves and blood vessels is important for preventing and treating the complications of diabetes. Neurological dysfunction and cell necrosis (including endothelial cell death) in diabetic patients is caused by complex events associated with an imbalance in diabetes metabolism ((ii))Neuron.2017Mar 22; 93(6):1296-1313). Specifically, hyperglycemia, dyslipidemia, and/or insulin resistance potentiate activation of the pathways such as polyols, glycosylation end products (AGEs) and the like, as well as loss of insulin signaling, ultimately contributing to impairment of mitochondrial function, inflammatory responses, and oxidative stress. Numerous studies have demonstrated that impairment of mitochondrial physiological function, reduction of respiratory chain function (decreased ATP levels) and increased ROS production are important triggers in a range of diabetic complications, including nephropathy, cardiomyopathy and neuropathy (a) ((ii))Diabetes.2008 Jun；57(6):1446-54；Handb Clin Neurol.2014；126:353-77；Neurobiol Dis.2013 Mar；51:56-65.Circ Res.2016May 27; 118(11):1808-29.). Currently, control of blood glucose levels and treatment of upstream pathways such as PKC, polyols, PARP and hexosamine pathways have not been satisfactory to date, nor have antioxidant and anti-inflammatory properties been shown to be effective against diabetic neuropathy separately (Neuron.2017Mar 22; 93(6):1296-1313). It can be seen that targeting individual events of the complex events leading to diabetic complications or targeting a pathway in the upstream signaling network leading to a complex event is not sufficient to produce a significant efficacy in treating diabetic complications. In particular, acquired mitochondrial dysfunction has also been associated with many common diseases, such as cardiovascular disease; neurodegenerative diseases (including senile dementia, parkinson's disease and multiple sclerosis); metabolic disorders (including diabetes and obesity). Although mitochondria play a central role in human health and disease, treatment of defects in mitochondrial function has not been successful (Nat Rev Drug Discov.2013 Jun；12(6):465–483.；Sci Transl Med.2016 Feb 17；8(326):326rv3.)。

In conclusion, new ideas are needed to prevent and treat diabetic complications-either to correct metabolic imbalances, or to protect mitochondrial function and block inflammatory responses and oxidative stress. The search for new methods and new drugs that can effectively prevent and reverse mitochondrial dysfunction and metabolic disorders is of great significance for the prevention and treatment of diabetic complications and neurodegenerative diseases.

Disclosure of Invention

The invention aims to provide application of a panaxadiol saponin Rb component (called Rb component for short) in preparing a medicament for preventing and treating diabetes complications and metabolic disorder related diseases. The drug can correct the metabolic disorder of diabetes mellitus by taking independent blood sugar level as a mechanism of action and can enhance the mitochondrial function and the redox balance capability of a diabetic patient, and the drug can also treat mitochondrial damage caused by pathological factors other than diabetes mellitus and pathological symptoms related to the metabolic disorder. That is, the present invention provides a drug for preventing and treating diseases and pathological symptoms having metabolic disorders, particularly mitochondrial dysfunction, as a common pathological mechanism.

The medicine is various medicinal preparations prepared by taking a panaxadiol saponin Rb component (hereinafter referred to as Rb component) as a single active ingredient or being mixed with other active compounds and a pharmaceutically acceptable carrier. The Rb component mainly comprises five panaxadiol saponins (Rb)₁、Rb₂、Rb₃Rc and Rd), one or two or more of the five panaxadiol saponins can be selected as active ingredients; the other active compounds include various substances that lower blood glucose levels, traditional Chinese medicine active substances, natural products, synthetic compounds and in vivo active substances. Or the independent preparation of the Rb component can be used in combination with the hypoglycemic agent in clinical use.

The total content of the five panaxadiol saponins accounts for 50-98% of the Rb component, preferably more than or equal to 90%, and the Rb component₁、Rb₂、 Rb₃The content of the five monomer compounds of Rc and Rd respectively accounts for 3-50% of the content of the Rb component, preferably more than or equal to 10-30%, but does not include the condition that the content of the five monomer compounds is more than 20% at the same time. The Rb component used in the present invention was prepared using the method provided in ZL 201210242928.1. It is understood by those skilled in the art that the application of the medicine prepared by using one or more than two of the five components as active ingredients in treating diabetic complications is also within the protection scope of the patent.

The metabolic disorder-related diseases include neurodegenerative diseases (Alzheimer's disease, vascular dementia, Parkinson's syndrome, multiple sclerosis and Huntington's disease) and neurological dysfunction diseases (including epilepsy, schizophrenia, depression) and peripheral and central neuropathic symptoms resulting from cancer treatment. The diabetic complications include various peripheral and central complications of diabetes.

The structural formula of the panaxadiol saponin is as follows:

ginsenoside Rb₁(ginsenoside Rb₁):R＝–D–glucopyranosyl

Ginsenoside Rb₂(ginsenoside Rb₂):R＝–L–arabinopyranosyl

Ginsenoside Rb₃(ginsenoside Rb₃):R＝–D–xylopyranosyl

Ginsenoside Rc (ginsenoside Rc) R (L-arbinofurosyl)

Ginsenoside Rd (ginsenoside Rd) R ═ H.

Some of the experimental findings of the present invention support the clinical use of the pharmaceutical preparations of the Rb component in the prevention and treatment of the disease.

The Rb component can obviously improve diabetic complications

The Rb component in combination with low doses of insulin significantly ameliorates the "more than three, one less" typical symptoms of diabetes, an effect that does not depend on a reduction in blood glucose levels. More than three, and less than one, the patient can have polyuria, polydipsia, polyphagia, physical strength and weight loss. This is not only a typical symptom of diabetes, but also a macroscopic response to metabolic disorders of diabetes. In particular, polyphagia reflects the body's inability to utilize glucose efficiently, the body attempts to gain energy by ingesting more food, and physical and weight loss further reflects energy deficits, the body producing energy by increasing consumption of fat and protein. The Rb component and the insulin can obviously reduce the water intake and the urine output of the mice with the type I diabetes mellitus when being used singly, and can recover the weight to a certain extent, and the Rb component and the insulin can obviously improve the 'more than three and less than one' of the mice with the diabetes mellitus when being combined. It is worth mentioning that the improving effect of the Rb component on "more than three, one less" is independent of blood glucose levels. For a long time, global diabetes treatment including the most developed countries and prevention of diabetic complications have been attempted by strictly controlling blood sugar levels, but practice has proved to achieve certain effects only on type 1 diabetic patients. Therefore, the drug effect of the Rb component for improving the diabetes without depending on the reduction of the blood sugar level has special significance for preventing and treating the diabetic complications, and the research result supports the special medical application of the Rb component and various hypoglycemic drugs for preventing and treating the diabetic complications.

The Rb component can also obviously promote the wound healing of diabetic animals and improve the motor balance ability and cognitive ability of the diabetic animals, which further supports the medical application of the Rb component in preventing and treating various complications of diabetes. Diabetic foot is a serious complication in the later stage of the disease, and the main reasons of the diabetic foot include 1) pain sense weakening or disappearance caused by diabetic peripheral sensory neuropathy, so that the patient loses self-protection capability and the foot is repeatedly damaged; 2) metabolic dysfunction and/or other pathological conditions result in a diabetic patient having diminished or even no repair of damaged tissues, including resistance to infection and chronic inflammation. Therefore, the drug effect of the Rb component in promoting the wound healing of the diabetic animals not only supports the medical application of the Rb component in preventing and treating diabetic feet, but also further points out the systemic improvement effect of the Rb component on diabetic metabolic disorder and other pathological states. The reduction of the motor balance ability of the diabetic not only reflects the diabetic peripheral sensory neuropathy, but also is an important reason for the easy falling and fracture of the diabetic. The loss of interest in newborns and the decrease in exploratory and cognitive abilities of diabetic patients reflect the central nervous dysfunction or decline of diabetes. Thus, the efficacy of the Rb component in improving motor balance, exploration and cognition in diabetic animals supports the prevention and treatment of diabetic peripheral neuropathy including motor sensory nerves and autonomic nerves and their associated complications including neuralgia, cardiovascular complications, complications of the digestive and urinary systems and the genitourinary systems, and central neuropathy and associated diabetic encephalopathy including hypomnesia, sleep disorders and psychobehavioral disorders.

The Rb component can aim at the core pathological mechanism of the diabetes complication and can fundamentally treat the diabetes and the complication thereof.

First, Rb can inhibit the activation of polyol and glycosylation metabolic pathways in cells of diabetic patients through an independent hypoglycemic mechanism, and this effect can be further potentiated in combination with low doses of insulin. The activation of the polyol and the glycosylation metabolic pathway is not only a result of the prolonged elevation of intracellular glucose levels in diabetic patients, but is also an important marker of metabolic disorders of diabetes and is directly involved in the development and progression of diabetic complications. It is known that the increase of glycosylated hemoglobin level not only reflects the increase of blood sugar level in the past but also reflects the increase of blood sugar level in the body for a long time, especially, excessive glycosylated end product (AGE) in the body of a diabetic patient can not only be crosslinked with protein to influence the performance of the protein, but also change the cell function by combining with a specific receptor, and can also trigger oxidative stress and inflammatory reaction to cause pathological changes of the body. In fact, AGE is closely related to diabetic complications such as diabetic nephropathy, retinopathy, neuropathy, atherosclerosis, insomnia, impotence, gangrene, gastroparesis (slow gastric emptying). Activation of the polyol metabolic pathway consumes NADPH in large amounts, resulting in decreased NO synthesis or decreased glutathione, with consequent decrease in blood flow to the blood vessels and production of large amounts of free radicals, causing nerve damage. In addition, since the nerve tissue does not contain fructokinase, fructose, which is an intermediate product of glucose passing through a polyol pathway, cannot be utilized, and as a result, a large amount of sorbitol and fructose are accumulated in nerve cells, resulting in intracellular hypertonicity and swelling, degeneration and necrosis of nerve cells. In conclusion, Rb can obviously improve the metabolic disorder state of diabetes and inhibit oxidative stress damage related to glycosylation and polyol metabolic pathway, thereby protecting the functions of various functional proteins and tissues and organs of a diabetic patient, and being beneficial to the Rb component to play a role in preventing and treating various complications of diabetes.

Second, the Rb component can reverse diabetic mitochondrial dysfunction and energy (ATP) deficiency. Type 2 diabetes is characterized by mitochondrial dysfunction, high reactive oxygen species production and low levels of ATP, and the Rb component not only completely reverses this metabolic profile, but in combination with low doses of insulin, increases the energy-producing function of diabetic mitochondria to levels greater than that of normal animals. The strong mitochondrial energy production function is beneficial to the exertion of various energy-consuming physiological functions, including daily walking activity, cardiac activity and central nervous activity with extremely high energy consumption, so that the body function and the life quality of the diabetic patient are improved.

Thirdly, the Rb component can treat diseases of the redox system of diabetes, greatly improve the antioxidant capacity of the diabetic animals, and further enhance the pharmacological significance of the Rb component in promoting the mitochondrial oxidative phosphorylation of the diabetic animals. Reduced Glutathione (GSH) and reduced coenzyme ii (NADPH) which reduces oxidized glutathione (GSSG) to GSH are the most important endogenous substances in the body to resist oxidative damage. In diabetic patients, the polyol pathway activates a large consumption of NADPH to convert glucose to sorbitol, thereby weakening the ability to reduce GSSG, and at the same time, there occurs a decrease in the levels of cysteine and glycine (two important substrates for GSH synthesis), and it can be seen that the decrease in GSH regeneration and de novo synthesis together result in low levels of GSH in type 2 diabetes. Consistent with lowering sorbitol levels in diabetic animals, the Rb component can significantly raise GSH and NADPH levels in diabetic animals, and its strength of action not only reverses the decrease in diabetic GSH, but increases them significantly above that of normal animals, without raising the levels of GSSG. These findings demonstrate that the Rb component significantly enhances the endogenous antioxidant capacity of diabetic animals, whereas insulin does not.

The Rb component can not only improve the ability of reducing GSSG by inhibiting a polyol pathway and reducing the consumption of NADPH; more importantly, the Rb component potentiates serine metabolism-mediated de novo biosynthesis of Glutathione (GSH) in diabetic animals, highlighted by elevated levels of key intermediates in various links of the synthetic pathway, including serine, cysteine, glutamine and glutamate; the Rb component increases NADPH levels, indicating that the Rb component can also increase nucleotide biosynthesis. In particular, Rb enhances NADPH regenerating ability of diabetes by regulating the action of the carbohydrate metabolism network. The carbohydrate metabolism activity is closely related to the regeneration of NADPH, wherein serine-mediated one-carbon unit metabolism is associated with not only nucleotide synthesis but also NADPH regeneration, and glucose-6-phosphate dehydrogenase (G6PD) in the pentose phosphate bypass, malic enzyme and NAD-dependent Isocitrate Dehydrogenase (IDH) in the tricarboxylic acid cycle play an important role in the regeneration of NADPH. However, these decreases in enzyme activity lead to a decrease in NADPH regeneration capacity and decreased NADPH levels in type 2 diabetes, and are associated with ROS generation, DNA fragmentation, lipid peroxidation, mitochondrial damage, and significant decreases in ATP levels. In particular, IDH is extremely sensitive to glycosylation, which leads to impaired IDH activity in type 2 diabetes, and in particular, is an important cause of NADPH reduction. The Rb component inhibits the polyol pathway, which acts to both reduce NADPH consumption and protect IDH-mediated NADPH regeneration activities, and also potentiates serine metabolism pathway-mediated NADPH regeneration.

In conclusion, the Rb component can regulate and control the metabolic network of diabetic animals from a plurality of joint links and pathway systems and can inhibit the damage of the glycosylation of diabetes mellitus to the metabolic network, thereby improving the oxidative phosphorization capacity and the antioxidant capacity of mitochondria. The action mechanism can reverse the metabolic disorder of diabetes, provides an action mechanism for the Rb component to improve more than three and less than one symptoms of diabetes and various complications, and also strongly prompts that the Rb component can prevent and treat other metabolic disorder-related diseases, including common neurodegenerative diseases.

The Rb component protects vascular endothelial cells from damage by high blood glucose levels

The vascular disease of diabetes is one of the common diabetic complications, which is also one of the major causes of death in diabetic patients. The deficiency of nutrition and nutrition caused by vasculopathy almost participates in the occurrence and development of all diabetic complications, in particular cardiovascular complications, cerebral apoplexy, nephropathy, blindness, neuropathy and diabetic foot. And damage to vascular endothelial cells underlies vascular lesions. The Rb component can significantly combat oxidative stress damage to endothelial cells caused by high sugar levels.

The Rb component also has obvious antagonistic action on the mitochondrial dysfunction and related pathological symptoms caused by non-diabetes, which supports that the pharmaceutical preparation of the Rb component prevents and treats diseases taking the mitochondrial dysfunction as a common pathological mechanism

In the mitochondrial respiratory chain inhibitor rotenone-induced non-diabetic neurodegeneration model, the Rb component protects the neuro-glial-vascular unit, inhibiting microglial activation and neuroinflammation mediated by peripheral inflammatory cell invasion. Mitochondrial dysfunction is known to be the core pathological mechanism of toxic and side effects of chemotherapy drugs, and in a chemotherapy drug paclitaxel-induced peripheral neuropathic pain model, the Rb component can completely block the occurrence and development of peripheral neuropathic pain. The results show that the Rb component has protective effect on the mitochondrial damage caused by different disease treatment factors, which strongly supports the prevention and treatment of diseases taking mitochondrial dysfunction as a common pathological mechanism by the medicinal preparation of the Rb component.

Mitochondrial dysfunction, oxidative stress injury, inflammatory response and vascular injury are common pathological mechanisms of diabetic neuropathy, cancer treatment-induced neuropathy and non-diabetic neurodegenerative diseases (including senile dementia, parkinson's disease, multiple sclerosis, etc.), with metabolic disorders being central as they can both induce and exacerbate oxidative stress injury, inflammatory response and vascular damage. Furthermore, a decrease in the oxidative phosphorylation capacity of mitochondria coupled with an increase in ROS production underlies intramuscular fat accumulation, insulin resistance and muscle dysfunction during aging. Therefore, the common pathological mechanisms strongly support the medical application of the pharmaceutical preparation of the Rb component in preventing and treating diabetic complications, neurotoxicity caused by chemotherapeutic drugs and neurodegenerative diseases (such as senile dementia, Parkinson's disease, multiple sclerosis and the like), and also support the medical application of the pharmaceutical preparation of the Rb component in preventing and treating other diseases related to metabolic disorder including psychobehavioral disorder.

Pharmaceutical preparations made with the Rb component include oral solid or liquid preparations, injectable preparations, sustained release preparations, controlled release preparations, targeted preparations, effervescent preparations or in ointments or creams for topical administration, also for example in the form of suppositories for rectal administration, also for nasal spray preparations. The oral preparation comprises oral tablet, buccal tablet, chewable tablet, pill, dripping pill, capsule, soft capsule, granule, oral liquid, syrup, emulsion, and mixture; the injection comprises small injection, large injection, powder for injection, emulsion, and suspension.

Generally, the pharmaceutical compositions of the present invention may be prepared in a conventional manner using conventional excipients or carriers known in the art.

Pharmaceutically acceptable solid excipients or carriers include: starch, corn starch, lactose, sucrose, sodium carbonate, calcium phosphate, phosphoglycol, calcium carbonate, alginic acid, microcrystalline cellulose, gelatin; pharmaceutically acceptable liquid carriers include, for example, sterile water, polyethylene glycol, non-ionic surfactants (e.g., hydroxypropyl cellulose, tween), hydroxypropyl- β -cyclodextrin and oils such as corn oil, peanut oil, sesame oil, olive oil or liquid paraffin; as appropriate to the nature of the active ingredient and the particular mode of administration desired. Adjuvants commonly used in the preparation of such pharmaceutical compositions may also include, for example, flavoring agents, coloring agents, preservatives (such as ethyl or propyl-hydroxybenzoate) and antioxidants such as vitamin E, vitamin C, BHT and BHA; pharmaceutically acceptable polymeric materials are also included in each of the above cases.

The tablets may be uncoated or coated to modify their disintegration and subsequent absorption of the active ingredient in the gastrointestinal tract or to enhance their stability and/or appearance, in which latter two cases conventional coating agents and methods known in the art may be used.

Pharmaceutical compositions for oral use may also be in the form of hard capsules wherein the active ingredient is mixed with an inert solid excipient, for example calcium carbonate, calcium phosphate, microcrystalline fiber, kaolin, or a vehicle for encapsulation, or in the form of soft capsules wherein the active ingredient is mixed with water or an oil, for example corn oil, peanut oil, sesame oil, olive oil, liquid paraffin, or a pharmaceutically acceptable polymeric material.

Pharmaceutical compositions in the form of a suitable for injection include sterile aqueous solutions, dispersions or sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. The carrier may be a solvent or dispersion medium or a coating medium, such as water, alcohol, suitable mixtures thereof and vegetable oils, as well as pharmaceutically acceptable polymeric materials.

The key points and the beneficial effects of the invention are as follows:

the invention provides a medicine which can aim at the core pathological mechanism of diabetic complication, namely a medicinal preparation of Rb component, and the medicine can fundamentally treat various diabetic complications. The pharmaceutical preparation containing the Rb component is completely different from the existing hypoglycemic drugs in the action mechanism, and the combined use of the pharmaceutical preparation containing the Rb component and the existing hypoglycemic drugs can generate the synergistic effect. The invention also provides a medicine which can prevent and treat various serious diseases based on the common pathology machine. Mitochondrial dysfunction induces oxidative stress and inflammatory responses that further exacerbate metabolic disorders and mitochondrial damage. The malignant chain reaction and waterfall amplification are not only the core pathological mechanism of diabetes complications, but also the common pathological mechanism of nerve toxic and side effects caused by neurodegenerative diseases and cancer chemotherapy. Therefore, the pharmaceutical preparation containing the Rb component can resist the damage of different pathogenic factors to mitochondria, enhance the function of mitochondria and enhance the function of an endogenous antioxidant stress system, so that the malignant chain reaction and waterfall amplification among mitochondrial dysfunction, oxidative stress damage and inflammatory reaction are prevented from the source, and the occurrence and the development of related diseases can be prevented.

Diabetic complications are the main causes of death and disability of diabetic patients, but up to now, there is no safe and effective medicine for preventing and treating diabetic complications in the global scope. Various neurodegenerative diseases represented by senile dementia and neurotoxic side effects caused by cancer treatment are also global intractable diseases. Therefore, the Rb component medicinal preparation provided by the invention can provide safer and more effective medicaments for patients with the diseases.

Drawings

FIG. 1 half-dose insulin and its combination with the Rb fraction cause a short-term blood glucose lowering in type I diabetic mice.

FIG. 2. Long-term treatment, insulin, Rb alone and in combination, did not lower blood glucose levels in type I diabetic mice.

FIG. 3 Rb component and its combination with insulin restore body weight in type I diabetic mice.

FIG. 4 Rb component and its combination with insulin reduce water intake in type I diabetic mice.

FIG. 5 shows that the Rb component and its use with insulin can reduce the amount of urine excreted by type I diabetic mice.

FIG. 6 Rb component and its use in combination with insulin can accelerate wound healing in diabetic mice.

FIG. 7 Rb component and its use in combination with insulin can promote wound healing in diabetic mice.

FIG. 8 Rb component and its use in combination with insulin improves the balance function in diabetic mice.

FIG. 9 Rb component and its use in combination with insulin improves cognitive and autonomy in diabetic mice.

FIG. 10 Rb component and its use in combination with insulin improves curiosity exploration in diabetic mice.

FIG. 11 shows that Rb component and its combination with insulin can reduce the sorbitol accumulation as the intermediate of erythrocyte polyol pathway and the glycosylated metabolic pathway marker glycosylated hemoglobin level in diabetic mice.

FIG. 12 Rb component and its use with insulin can increase ATP levels as an energy producing substance in diabetic mice.

FIG. 13 shows that the Rb component and its combination with insulin can increase Glutathione (GSH), a key substance of the redox system of diabetic mice, and cysteine (Cys), a raw material substance of the diabetic mice.

FIG. 14 Rb component and its use in combination with insulin improves the levels of NADPH, a key coenzyme in polyol metabolism in diabetic mice.

FIG. 15 Rb component reduces high sugar induced oxidative stress, reduces endothelial and nerve cell damage.

FIG. 16.Rb component is directed against astrocyte, microglial activation and nerve cell damage caused by mitochondrial damage.

FIG. 17 infiltration of Rb component into the brain against rotenone-induced peripheral immune cells in Parkinson rats.

FIG. 18.Rb component prevents and treats neuropathic pain induced by chemotherapeutic drugs (mouse thermo-sensitive test).

FIG. 19 shows that Rb component prevents and treats neuropathic pain caused by chemotherapeutic drugs (rat thermo-sensitive test).

Detailed Description

The present invention is further illustrated by the following specific examples, which should not be construed as limiting the scope of the invention to the following examples, which should be construed as within the scope of the invention, and any alterations in the field which are made in accordance with the teachings of the present invention should be within the scope of the invention.

Example 1 Effect of the Rb fraction on blood glucose levels and metabolic characteristics (polydipsia, polyuria, body wasting symptoms) in type I diabetic mice

The method comprises the step of carrying out intraperitoneal injection (i.p.) once by using streptozotocin (150mg/kg) to cause a type I diabetes model of Kunming mice, wherein the increase of blood sugar level of the model animals is caused by the deficiency of insulin level due to the damage of islet beta cells, and diabetic complications caused by the model are related to persistent hyperglycemia and insulin signal loss. This provides an opportunity to examine the effect of the test drug on the metabolic state and complications of diabetes and its mechanism of action from multiple perspectives. Firstly, the model is used for observing the improvement effect of the Rb component alone or together with insulin on the pathological characteristics of polydipsia, diuresis, body emaciation and the like of the type I diabetic animals. The 35 qualified mice were divided into a normal animal control group, a type I diabetes model control group, an insulin group (2.5IU/kg), and an Rb group (40mg/kg) treatment group, and the combination of the insulin and Rb groups (2.5IU/kg +40mg/kg) was 7 mice per group. Insulin is injected subcutaneously in the abdomen once a day, the Rb component is administered by intragastric administration once a day, and the combined group is injected intragastric in half an hour after the subcutaneous injection of the insulin until the experiment is finished, and the same amount of physiological saline is injected intragastric in a control group. The weight, water intake, and moisture level of the animal bedding were recorded daily for each group of animals and were photographed to reflect the animal's urine output. In addition, blood glucose was measured once a week with blood glucose strips, and the dynamic changes in blood glucose levels that may occur over the course of treatment were monitored: blood glucose levels were measured 2 hours after dosing to understand the direct effect of drug treatment on blood glucose levels; blood glucose levels were measured 24 hours fasting during long-term dosing to see the steady changes in blood glucose levels that the drug may produce.

As a result:

the Rb component does not affect blood glucose levels in type I diabetic animals

As shown in the figure 1-2, after the STZ is injected into the model in large dose, the blood sugar level of the model group animals is more than 16.7mmol/L, which proves that the model is successfully made. First and multiple administrations 2Hourly and clinical half dose of insulin (2.5IU/kg) and the combination of the insulin and the 40mg/kgRb component can rapidly reduce the blood sugar level of mice with type I diabetes mellitus^***P<0.001vs prior to administration), but administration of the Rb component alone did not lower blood glucose levels. To determine whether the blood glucose levels of the animals of each administration group had improved after 2 weeks of treatment. Fasting blood glucose levels were determined 24 hours after the previous dose. As a result, it was found that no significant difference was observed in blood glucose levels of the animals of each administration group as compared with those before the treatment, and no significant difference was observed in blood glucose levels of the model animals (P)>0.05 vs) indicating that the Rb component, insulin and both cannot restore lost islet function in type I diabetic animals (fig. 1, fig. 2).

It is particularly noted here that since the Rb component has neither an acute nor a chronic effect on blood glucose levels, this sets the prerequisite that the Rb component does not alter blood glucose levels in diabetic animals. That is, any drug effect of the Rb component on diabetic animals does not depend on the change of blood sugar level, and the potential effect characteristic of the Rb has special significance for preventing and treating diabetic complications (see the following discussion for details).

The Rb component can improve the metabolic characteristics of type I diabetes and has better curative effect when combined with insulin

As shown in FIGS. 3 to 5, the diabetic model animals exhibited the typical symptoms of "more than three and less than one" of the metabolic disorders of clinical type I diabetic patients, and the mice in the diabetic model group were significantly lower in body weight than the normal group, while the water intake and urine output were significantly higher than those of the normal animals.

Both the Rb fraction and insulin alone reduced water intake in diabetic animals (b)⁺⁺P<0.01vs model), but no improvement in weight loss of diabetes was observed, and the combination of both significantly improved various indices (fig. 3-5). Within 24 hours, the bedding for normal animals was substantially dry, while the bedding for model group animals was 80% wet. The animal litter moisture was significantly improved in each of the dosed groups compared to the model group, with the feed of animals with the combination of insulin and Rb components being the most dry (fig. 5). It is emphasized that the improvement in the various indices increases with the duration of the treatment, which indicates that these indices are relevantThe targeted improvement is a stable drug effect produced by the combined treatment of the Rb component and insulin, which is a pharmacodynamic response to metabolic state deterioration. Therefore, research results support the medical application of the Rb component in preventing and treating diabetes by combining with insulin and other hypoglycemic drugs.

In order to further observe the possible effects of the Rb component and the combination of the Rb component and the hypoglycemic agent on the prevention and treatment of the diabetic complications, the research contents of the examples 2 to 8 are added in the test system of the example 1.

Example 2 the Rb component promotes the lesion repair function in diabetic animals

The method comprises the following steps: in order to reveal the potential medicinal value of the Rb component and the combined treatment of the Rb component and the hypoglycemic agent on the prevention and treatment of the diabetic foot, the wound repair condition of each group of animals is observed by utilizing a diabetic chronic comprehensive wound model. After 5 weeks of treatment, animals were anesthetized with pentobarbital sodium, and then the longer mouse hairs were subtracted with electric clippers and the depilatory cream had the back hair removed. Then, the skin on the back is disinfected by iodophor, a 1.5cm multiplied by l.5cm full-layer skin injury wound surface is formed by scissors in the back under the aseptic condition, the wound surface is cut to a subcutaneous fascia, and the skin around the wound is covered by gauze after being disinfected by alcohol. After the operation, the mice are fed in a single cage, and are freely fed and drunk, and the animals of each group continue to be treated or treated by the corresponding medicines. The wound healing conditions were observed on

days

1,3, 6, 9, 12, and 15 after molding (also after wound treatment), respectively, and the wound areas were counted.

As a result: as shown in the figure, the wounds of the animals in the administration group were in rapid healing with the prolonged administration time. The Rb component, the insulin alone and the combination of the two always keep a higher healing rate, wherein the Rb component alone is better than the insulin alone, and the combination of the two has the fastest healing rate. By the 10 th day of administration, the wound healing rate (78 + -7.3%) of the Rb component and insulin combined group was significantly different from that of the model group (53 + -7.2%) (⁺⁺P<0.01vs model) (fig. 6).

In the later period of wound healing (day 14 of administration), the skin condition of the wound of the mice was observed to find that the skin of the normal group and the skin of the combined group are basically healed, hair on the surface of the animal grows out to cover the wound, but the model group and the insulin group are not completely healed and are in a wound exposure and scab state (figure 7). Indicating that the Rb component improves the speed and state of wound healing in diabetic animals.

Example 3 Rb component improves motor balance function in diabetic animals-rotarod experiment

The method comprises the following steps: in order to reveal the potential medicinal value of the Rb component and the combined treatment of the Rb component and the hypoglycemic agent on the prevention and treatment of the diabetic peripheral neuropathy, the motor balance capability of each group of animals is investigated. After 8 weeks of treatment with the drug, the motor balance ability of each group of animals was measured with a rotarod apparatus. The animal must maintain a proper movement speed in order not to fall off the rotating rod, so that the longer the animal is maintained on the rotating rod, the stronger the movement balance ability and the motor muscle ability of the animal are indicated. 12 hours after dosing, the animals were placed on a fatigue meter with a speed set at 15 rpm for 2 minutes of the experiment, and the meter was started to uniformly accelerate to the set speed. An adaptation time of 15s was given first and it was ensured that the head of the mouse was facing in the opposite direction to the rotation of the rotarod and that the limbs were all attached to the rotarod. The timing was started when the instrument was started, and when the mouse dropped from the rotating bar, the instrument automatically stopped timing and displayed the time the mouse moved on the rotating bar, which the experimenter recorded.

As a result: as shown in FIG. 8, the time of movement on the rotarod (27.6. + -. 7.3) was significantly less in the diabetic model group than in the normal group of mice (99.2. + -. 37.1) (. times.P)<0.01vs model), indicating that the motor balance ability of the diabetic animals is significantly reduced. Insulin alone (19.7 + -14.1) did not improve the motor ability (P) in type I diabetic mice>0.05vs model), the Rb component alone (35.8 + -25.4) showed a tendency to improve the motor balance ability of mice, while the combination of the two (44.8 + -11.7) significantly improved the motor balance ability of diabetic animals. (⁺P<0.05vs model).

Example 4 the Rb component improves cognitive function-novel object recognition in diabetic animals

The method comprises the following steps: in order to further reveal the potential medicinal value of the Rb component and the combination of the Rb component and the hypoglycemic agent for preventing and treating the diabetic central neuropathy, a novelty identification test is used for observing the identification capability of each group of animals to novelty and investigating the movement time of the animals in the exploration activity to reflect the autonomous movement capability of the mice. In the novelty identification test, the greater the number of times or the longer the animal has been exposed to fresh objects, the better the animal's short term memory, and the greater the interest in fresh objects, and thus the novelty identification test can determine the level of cognition and memory in the animal. At 8 weeks after the administration of the treatment, novelty recognition tests were performed on each group of experimental animals. The first part of the experiment was an adaptation phase, where two identical things (object 1 and object 2) were placed in opposite positions in a box and the mice were allowed to adapt for 10min after placement. After an interval of 1h, a second partial test phase was carried out, in which object 1 was unchanged (old), object 2 was changed to object 3 (new), and then the mouse was again placed for 3 min. Each mouse explored the use of alcohol to wipe the object and box to eliminate the smell of the mouse staying on the object and box. The number of times the mouse was explored on each object was recorded, as well as the time of the exploration movements of the mouse in the open field of the box. Reflecting the exploration and short-term memory capacity of the mouse on the new object by using a new object priority index (the new object priority index is the frequency of exploring the new object/(the frequency of exploring the old object + the frequency of exploring the new object)); and the movement time of the mouse exploration process reflects the autonomous movement capability of the mouse.

As a result: as shown in fig. 9, compared to normal animals (0.82 ± 0.07), the preference for new animals in the model group was significantly reduced (0.37 ± 0.29) (. P <0.05 vs. model), and the time of autonomic activity in mice (70.8 ± 25.4) was also significantly reduced relative to normal animals (143.0 ± 24.9) (. P <0.01 vs. model). The results of the test indicate a decrease in memory, search and mobility in the diabetic animals, consistent with a decrease in curiosity, cognitive function and motor ability in the diabetic.

Insulin treatment fails to improve the memory cognitive ability (P) of type I diabetic mice>0.05vs model), the Rb component alone treatment tended to increase the preference index for new events (0.61 + -0.35) and significantly increased the exploration time in the open field trial (121 + -36) for diabetic animals (1: (0.05 vs model)⁺P<0.05vs model), the combination therapy of the Rb component and insulin can simultaneously and obviously improve the priority index of diabetic animals to new things(0.74±0.16)(⁺P<0.05vs model) and search times in the open field test (136.2. + -. 18.2) ((S)⁺⁺P<0.01vs model) and close to normal animals. The results of the study demonstrate that the Rb component significantly improves cognitive memory, exploration and exercise in diabetic animals, particularly when administered in combination with insulin (figure 9).

Example 5 the Rb component improves the free exploratory ability of diabetic animals-A well plate experiment is also known as the hole plate experiment

The method comprises the following steps: to further determine the Rb acting moiety and its effect on improving the exploratory ability of diabetic animals in combination with insulin, the exploratory ability of each group of animals was further determined using a well plate assay. In the plate experiment, the more times the animal drills the hole, the stronger the exploration ability of the animal is. After 8 weeks of administration, the mice were subjected to the well plate experiment. The experiment requires dim light. The hole plate experiment device is a square board with the size of 76cm multiplied by 76cm, 16 round small holes with the diameter of 5cm and equal intervals are arranged in the board, a hollow black box with the height of about 5cm is arranged below the holes, opaque transparent glass plates with the height of 50cm are arranged around the board to isolate the influence of the external environment, a mouse is placed in the center when the experiment starts, timing is started at the same time, and an observer records the times of exploring the black hole by the mouse within 3 min. The criteria recorded are the number of times the mouse has penetrated the head into the hole.

As a result: as shown in FIG. 10, the number of black hole searches in diabetic model animals (15.8. + -. 4.08) was significantly less than that in normal animals (34.86. + -. 3.34), indicating a decrease in the exploratory ability of diabetic animals. The single treatment of the Rb component (21.2 +/-4.76) and the insulin (22.61 +/-0.14) can increase the exploration times of diabetic animals to a certain extent, but the statistical significance is not achieved, and the combined treatment of the Rb component and the insulin (29.2 +/-2.86) can obviously increase the exploration times of the diabetic animals (the treatment of the Rb component and the insulin (1, 2 +/-4, 76))⁺⁺⁺P<0.001vs model) and levels near those of normal animals. Combining the results of the studies of examples 3 to 5, it was demonstrated that the Rb fraction improves neurocognitive function and motor function in diabetic animals, and the efficacy is better in combination with insulin.

Discussion and summary (examples 1-5) — effects of Rb component-produced non-blood glucose level dependent diabetes improvement of "more than three, less than one" symptoms, motor balance ability and cognitive ability, and promotion of wound healing in diabetic state not only support its medicinal use for prevention and treatment of diabetes-related complications, but also indicate that Rb component can comprehensively improve diabetic metabolic disorders.

Diabetic foot is a serious complication in the later stage of the disease, and the main reasons of the diabetic foot include 1) pain attenuation or disappearance caused by diabetic peripheral sensory neuropathy, so that the patient loses self-protection ability and the foot is repeatedly damaged; 2) metabolic dysfunction and/or other pathological conditions result in impaired or even abolished repair of damaged tissues in diabetic patients, including resistance to infection and chronic inflammation. Therefore, the drug effect of the Rb component in promoting the wound healing of the diabetic animals not only supports the medical application of the Rb component in preventing and treating diabetic feet, but also further points out the systemic improvement effect of the Rb component on diabetic metabolic disorder and other pathological states.

The reduction of the motor balance ability of the diabetic not only reflects the diabetic peripheral sensory neuropathy, but also is an important reason that the diabetic easily falls down and fractures. The loss of interest in newborns and the decrease in exploratory and cognitive abilities in diabetic patients reflect the dysfunction or decline of the diabetic central nervous system. Thus, the efficacy of the Rb component in improving motor balance, exploration and cognition in diabetic animals strongly supports the prevention and treatment of diabetic peripheral neuropathy including motor sensory and autonomic nerves and their associated complications including neuralgia, cardiovascular complications, complications of the digestive and urinary systems and the endocrine and reproductive systems, and central neuropathy and associated diabetic encephalopathy including hypomnesia, sleep disorders and psychobehavioral disorders.

To further confirm the overall improvement effect of the Rb component on diabetic metabolic disorders, further studies of examples 6 to 8 were conducted in the same experimental system as described above. Oxidative stress and chronic low grade inflammation are not only characteristic of diabetes, but also play an important role in the pathogenesis of the disease and the accompanying vascular and other complications. ROS production is associated with hyperglycemia and metabolic disorders such as impaired antioxidant function and impaired antioxidant activity caused by activation of polyol pathways and protein glycosylation and other mechanisms. Chronic exposure to oxidative stress conditions can cause chronic inflammation and fibrosis in a range of tissues, leading to the development and progression of disease states in related tissues. It can be seen that the observation of ROS levels, antioxidant systems including Glutathione (GSH) and NADPH levels, and their biosynthetic status in diabetic animals reflects both the metabolic derangement status of the disease and the extent of the disease. Similarly, observation of the effect of drugs on these indicators can reflect not only the action of drugs on correcting metabolic disorders of diabetes, but also the action mechanism of drugs on preventing and treating diabetic complications from an important perspective.

Example 6 treatment of diabetic complications by the Rb component through the non-glucose dependent regulation of the diabetes related polyols and the glycosylation metabolic pathway

The method comprises the following steps: after 8 weeks of drug treatment, 24 hours after the last administration, the mouse orbit was bled and centrifuged, and the packed red blood cells were removed to determine the sorbitol content and the glycated hemoglobin content, in order to examine the activation of the polyol and glycosylation metabolic pathways and the effects of the Rb component in diabetic animals. The level of protein glycosylation is expressed as absorbance value (OD).

As a result: as shown in fig. 11, sorbitol accumulation (15.98 ± 0.66mmol/L) in erythrocytes of model mice was significantly higher than that of erythrocytes in normal group (10.12 ± 2.21mmol/L) (. P <0.01 vs. normal group), and glycosylated hemoglobin level (35.64 ± 5.73) was also significantly higher than that of erythrocytes in normal animals (16.59 ± 6.56) (. P <0.01 vs. normal group). The research data is consistent with sorbitol pathway and glycosylation activation phenomenon of diabetic patients, and the model animal well simulates metabolic disorder state of diabetes. The insulin treatment can obviously reduce the sorbitol accumulation and improve the glycosylated hemoglobin level to a certain extent, the Rb component treatment also has the same pharmacological action, and the combined efficacy of the two is further enhanced, particularly the combination of the two can reduce the sorbitol level (9.02 +/-2.77) to the level of normal animals (+ + P <0.01vs model group). Based on the fact that polyol pathway and protein glycosylation activation are typical indexes of the diabetic metabolic disorder state, the research results prove that the Rb component can improve the diabetic metabolic disorder state particularly by combining treatment with insulin, and therefore, the comprehensive regulation and control effect of the Rb component on the diabetic metabolic disorder is deduced. The medical application of the Rb component in preventing and treating the diabetic complications is supported, and particularly the medical application of the Rb component in preventing and treating the diabetic complications by combining with hypoglycemic drugs.

Example 7 the Rb component can improve mitochondrial energy metabolism status in diabetic mice

The method comprises the following steps: mitochondrial dysfunction and reduced ATP levels are common pathological features of diabetes and are also important causes for further initiation and exacerbation of diabetic complications. Therefore, in a diabetes model, ATP levels may reflect both the effect of the drug on the metabolic state and the mechanism of action for preventing and treating diabetic complications. After 8 weeks of administration, muscle tissue was homogenized, and the homogenate was used to determine the level of energy metabolites in mouse muscle by high performance liquid mass spectrometry. The measurement results were converted to normal group units of "1".

As a result: as shown in FIG. 12, the energy metabolites ATP levels (0.52. + -. 0.32) and ADP levels (0.61. + -. 0.35) in the diabetic mice were reduced to some extent compared with those of the normal animals ATP (1.00. + -. 0.41) and ADP (1.00. + -. 0.38). Insulin treatment can significantly improve ATP level (1.22 +/-0.30) of diabetic animals⁺P<0.05vs model group), did not significantly affect ADP levels, suggesting that insulin can increase mitochondrial oxidative phosphorylation levels, i.e., promote conversion of ADP to ATP. The research result accords with the physiological function of insulin. Interestingly, the Rb component significantly upregulated both ATP (0.88. + -. 0.59) and ADP (1.48. + -. 0.66) ((R))⁺⁺P<0.01vs model group) and the magnitude of upregulation tended to exceed that of normal animals. And combined treatment with insulin and Rb component further increases ATP (2.92 + -0.37) and ADP (2.72 + -0.30) levels in diabetic animals to levels above those in normal animals ((2.92 + -0.37))⁺⁺⁺P<0.01 vs. model set<0.01vs normal control), indicating synergy between the two. It is reasonable to assume that an increase in ADP levels provides a more abundant substrate for mitochondrial oxidative phosphorylation, i.e., simultaneous increases in ADP and ATP levels indicate mitochondrial oxidative phosphorylation activityIncreased motility and mitochondrial productivity capacity. In conclusion, the Rb component not only completely reverses the impaired oxidative mitochondrial phosphorylation activity caused by insulin deficiency, but also increases mitochondrial energy production capacity in diabetic animals. Thus, the Rb component can completely reverse ATP deficiency in diabetic patients enough to provide energy support for the diabetic patients' work, daily activities, and functional maintenance and execution of various organs, and can avoid various pathological events and vicious cycles induced by energy deficiency. The synergistic effect of the insulin on the Rb component not only accords with the theory that the synergistic effect can be generated by combining two pharmacological active substances with different action mechanisms, but also supports the medical application of the Rb component and the hypoglycemic drug for preventing and treating the diabetic complications.

Example 8 the Rb component can enhance the endogenous redox potential of diabetes and thus can enhance the resistance to oxidative damage from external environmental sources

The method comprises the following steps: glutathione (GSH) and NADPH play a crucial role in maintaining redox homeostasis in vivo. After 8 weeks of administration, muscle tissue was homogenized, and the homogenate was used to determine the levels of Glutathione (GSH) and oxidized glutathione (GSSG), which are key substances of the mouse muscle redox system, and glycine (Gly) and cysteine (Cys), which are glutathione precursor substances, by high performance liquid chromatography-mass spectrometry. NADPH and NADP + levels were also measured to understand the large consumption of NADPH and the body redox state that the sorbitol pathway activates for possible metabolism and the role of the Rb component. The measurement results were converted to "1" in the unit of the normal group.

As a result: as shown in fig. 13, GSH levels (0.65 ± 0.43) and the synthetic raw material cysteine levels (0.66 ± 0.20) were significantly reduced in diabetic mice compared to normal animals, while no significant change was seen in GSSG. It can be seen that the function of synthesizing GSH is reduced in diabetic mice. With respect to the NADPH level (FIG. 14), the model group showed a tendency to decrease (0.83. + -. 0.56 vs. 1.00. + -. 0.52) compared to the normal group, but the level of NADP + was significantly lower than that of the normal animals (0.69. + -. 0.27 vs. 1.00. + -. 0.63), indicating that a decrease in the NADP + reducing ability was likely to occur in the diabetic metabolic disorder state. Taken together, the results of the study demonstrate the ability of diabetic animals to synthesize and use antioxidant substancesAre all damaged. The antioxidant capacity of diabetic animals is further challenged by NADPH reduction caused by the activation of the polyol pathway. Treatment with the Rb component can completely reverse this dysfunction of the redox system in diabetic animals, whereas insulin does not. For GSH level, the animal treated by Rb component and insulin are all obviously higher than that of diabetic animal: (⁺⁺⁺P<0.01vs model group) even exceeded normal animals (Rb fraction 1.78 ± 0.41, combination treatment 1.40 ± 0.32), Rb fraction treatment and its combination with insulin also significantly increased Cys levels ((r)⁺⁺⁺P<0.01vs model group), but each treatment group did not affect GSSG levels in diabetic animals. In terms of NADPH and NADP + levels, the Rb component treated animals were significantly higher than those of NADPH (1.58 + -0.36) and NADP + (1.09 + -0.64) of diabetic mice, but the Rb component in combination with insulin treatment could further significantly increase the NADP + levels ((1.88 + -0.68) of diabetic animals to more than normal animals.

Example 9 the Rb component reduces high sugar induced oxidative stress and endothelial cell injury

The method comprises the following steps: to examine the effect of Rb component in reducing high-sugar induced endothelial cell damage, human umbilical cord vascular endothelial cells Endo cultured for 24 hours were switched to high-glucose (150mM) containing medium, respectively, to establish a high-sugar damage model of endothelial cells. The experimental setup groups included a normal control group (CON) without any drug treatment, a high sugar culture control group and an Rb group-treated high sugar condition culture group cultured under normal conditions. Adding Rb components with different concentrations while starting high-sugar culture, continuously culturing for 72 hours, and then measuring the cell viability by an SRB method; ROS and mitochondrial membrane potential were measured and incubated for 24 hours. Measuring the Reactive Oxygen Species (ROS) in the cells by using a DCFH-DA probe method; the cellular Mitochondrial Membrane Potential (MMP) was measured by JC-1 fluorescence probe method, and the value of MMP was expressed by the ratio of red fluorescence intensity to green fluorescence intensity, and the data from the study can illustrate the effect of Rb component on the resistance to oxidative stress caused by hyperglycemia.

As a result: as shown in fig. 15, the Rb component does not significantly affect the viability of cells cultured under normal conditions, and high concentrations of glucose result in about a 12% decrease in the viability of Endo cells, while the Rb component can completely counter this decrease. In the intracellular active oxygen detection, ROS of the vascular endothelial cell ENDO high-sugar model group is increased by 83.6 percent compared with that of the normal control group<0.001), the Rb component was able to reverse the rise in ROS after intervention in the high-sugar model, i.e. ROS production was significantly lower than in the model group (b) ((b)^###P<0.001) and corresponds to the blank group. In mitochondrial membrane potential assay, MMP was reduced by 28.9% (. x.) P in endothelial cell ENDO high carbohydrate model group compared to blank group<0.001), the Rb component reverses the reduction of MMPs after intervention in the high carbohydrate model, i.e. MMP levels are close to the blank group. The research result shows that the culture condition with high sugar can cause the stress injury of vascular endothelial cells, and the Rb component can reduce the oxidative stress and the injury of the endothelial cells caused by the high sugar.

Discussion and summary (examples 6-9) -the Rb component can prevent the development of the core pathological mechanism of diabetic complications and can fundamentally prevent and treat diabetic complications

Research results prove that Rb has pharmacological action which is not possessed by insulin, and the Rb not only can correct metabolic disturbance characteristics of type I diabetes, but also can strengthen mitochondrial function and endogenous redox balance system function in a diabetic state and reach the level exceeding that of a strong animal, so that strong energy support can be provided for a diabetic patient, and various clinical symptoms of the diabetes caused by energy deficiency can be completely reversed; more importantly, the traditional Chinese medicine composition can prevent and treat oxidative stress and inflammatory reaction caused by mitochondrial dysfunction and subsequent vicious circle and waterfall amplification of the three, thereby fundamentally preventing and treating various complications of diabetes.

Example 10 the Rb component can combat against astrocyte, microglial activation caused by mitochondrial damage, neuronal cell damage and peripheral immunoinfiltration caused by blood brain barrier damage

The method comprises the following steps: mitochondrial respiratory chain inhibitors by subcutaneous injectionAn animal model of rotenone induced mitochondrial dysfunction in rats. Models are also used in models of parkinson's disease, since dopaminergic neurons are particularly sensitive to mitochondrial damage and the model animals show symptoms of parkinson's disease. The injection was started with a low dose and then 25% of the dose was escalated every 5 days, and the single administration of the daily dose was changed to an average of morning and evening doses. The specific method comprises the following steps: the first 5 days of modeling dose was 0.5mg/kg, the second 5 days of modeling dose was increased to 0.625mg/kg, the 3 rd 5 days of modeling dose was 0.75mg/kg, each dose was 0.05mL/100g, morning (8: 00, am) and evening (20: 00, pm) once. The normal control group was given an equal volume of sunflower oil each time. In the experiment, the injection of rotenone is stopped when the model animal has behavioral symptoms of grade 4 or above, and if the rats have animals which do not reach the grade 4 behavioral manifestations after the third 5 days, the model is continuously manufactured according to the administration dosage of the third 5 days. Normal control group

Model group (Rotenone), Rb group 40(Rb40, 40 mg/kg/day). The administration is carried out 60 minutes before the administration of the derris trifoliata according to the set dose, wherein each dose is half of the total daily dose, and each administration volume is 0.2mL/100 g. Normal control group and rotenone model control group were given equal volume of physiological saline. Neuroinflammation in the brain, and the health status of sensitive neurons (nigrostriatal pathways) were observed 3 weeks after administration. Brain tissue was fixed by heart perfusion with 4% paraformaldehyde. The brain tissue was coronal sectioned and subjected to relevant immunohistochemical experiments to investigate the dopamine neuron markers Tyrosine Hydroxylase (TH), the PV interneuron marker Parvalbumin (PV), the astrocyte marker Glial Fibrillary Acidic Protein (GFAP), the microglial marker (Iba-1) and the mature macrophage marker F4/80.

As a result: the increase of astrocytes and microglia soma and the thickening and shortening of axons appear in the brain of the model group rats, which indicates that the two cells are in a stress activation state (figure 16), thereby generating neuroinflammation. In particular, large areas of vascular destruction occurred in the brains of the model animals, and a large number of peripheral macrophages infiltrated into the brains through the damaged vessels (fig. 17). Consistent with this, model animals developed impairment of the nigrostriatal dopamine pathway, manifested by a decrease in positive intensity of TH staining, and loss of striatal PV interneurons. The Rb component can remarkably resist various pathological changes of model animals and even maintain the normal state. Research results show that the mitochondrial inhibitor can activate neurogenic inflammation in which microglia and astrocytes participate in brain, and can also destroy blood vessels to cause peripheral immune cells to invade the center, so that neurogenic inflammation is further aggravated and nerve degeneration and damage are caused; the Rb component can be used for remarkably resisting neuroinflammation, cerebrovascular injury and nerve injury induced by mitochondrial inhibitor. This supports the Rb component in protecting mitochondria and preventing and treating related diseases caused by mitochondrial damage or dysfunction.

Example 11 prophylactic Effect of Rb fraction on model of paclitaxel-induced neuralgia in mice

The method comprises the following steps: in the research, an ICR female mouse peripheral neuralgia model with the weight of 20-24g is induced by adopting a method of carrying out intraperitoneal injection on paclitaxel (2.8mg/kg) for 4 times every other day (

days

1,3,5 and 7), and the model is used for observing the prevention effect of the Rb component on peripheral neuralgia caused by chemotherapeutic drugs. Among the numerous antitumor drugs, the natural plant antitumor drug accounts for the largest proportion, and in the first year of 2007, the taxol in the plant antitumor drug occupies the first place of the market in 44.1% of the share, and the taxol antitumor drug has become a first-line drug for human to resist malignant tumors. The dose-limiting toxicity of paclitaxel is mainly neurotoxicity and myelosuppression, the latter is successfully overcome by applying granulocyte colony stimulating factor, but the neurotoxicity which shows neuropathic pain still troubles clinicians all over the world, because the pain caused by chemotherapy is insensitive to any analgesic drugs used clinically at present, a part of patients are forced to be reduced until stopping taking medicine, the chemotherapy effect is seriously influenced even the chemotherapy fails, and the part of paclitaxel chemotherapy pain is not rapidly stopped because of stopping taking medicine, and is usually prolonged for months or years, thus the life quality of tumor patients is seriously influenced. As can be seen, paclitaxel-induced peripheral nerve pain is representative of pain after cancer treatment, and animal models of pain induced by paclitaxel violet are also representative.

The experiment was conducted by screening mice with relatively uniform heat-sensitive reaction by hot plate method. The eligible 21 mice were divided into a blank control group (saline group, ip), a paclitaxel model group and an Rb group (40mg/kg, ig) prophylactic treatment group, 7 mice per group. The Rb component is administered by gavage once a day, and the Rb component is administered for 2 hours every time paclitaxel is administered. The Rb fraction was continued after paclitaxel withdrawal until the end of the experiment.

The thermo-sensitive response of the mouse hind paw was measured with a hot plate experiment (52 ℃. + -. 0.3) each time at 2-4 pm. The two lateral hind paws of the mouse are placed on a hot plate instrument, when the animal feels pain caused by thermal stimulation, the animal licks the hind paw or grabs after retraction, the latency period of licking the hind paw or grabs after retraction is recorded, the shorter the latency period is, the lower the pain threshold value is, and the pain threshold value prolonged to the paclitaxel animal shows that the paclitaxel animal has an antagonistic effect on the neuropathic pain induced by the chemotherapeutic drugs. The Rb fraction was continued after the paclitaxel injection was terminated and the thermosensitive response of each group of animals was continued to be measured.

As a result: the Rb component has a remarkable inhibiting effect on peripheral nerve pain of mice caused by paclitaxel. As shown in figure 18, paw withdrawal latencies were comparable in the three groups of animals prior to paclitaxel administration, and were significantly shorter in the model group than in the blank control group at days 7, 9, 11, and 13 following paclitaxel administration, indicating that paclitaxel induced significant peripheral neuropathic pain (P <0.01, P < 0.001); the latency period was significantly longer in the Rb group at day 7 and later time points than in the model group (. times.p <0.01,. times.p < 0.001). The above experimental results support the clinical use value of the Rb component in preventing and treating pain after cancer treatment, especially peripheral neuropathic pain caused by chemotherapeutic drugs.

Example 12 prophylactic Effect of Rb fraction on model of paclitaxel-induced neuralgia in rats

The method comprises the following steps: the research adopts a method of carrying out intraperitoneal injection on paclitaxel (2mg/kg, ip) for 4 times every other day (

days

1,3,5 and 7) to induce an SD male rat neuralgia model with the body weight of 300-. A heat-sensitive screening was performed on 14 rats by the hotplate method, and the 12 qualified rats were divided into paclitaxel and Rb (30 mg/kg, ig) prophylactic treatment groups of 6 rats each. The Rb component is administered by gavage once a day, and the Rb component is administered for 2 hours every time paclitaxel is administered.

The thermo-sensitive response of the rat hind paw was determined with a hot plate experiment (52 ℃. + -. 0.3) each time at 2-4 pm. The rats are placed on a hot plate instrument at both sides of the hind paw, when the animals feel pain caused by thermal stimulation, the animals retract and grab, the time (seconds) from the contact of the hind paw of the rat with the hot plate to the grabbing before retraction is recorded, namely the heat response latency, the shorter the latency is, the lower the pain threshold is, and the prolongation of the pain threshold of the paclitaxel animals indicates that the paclitaxel animals have resistance to neuropathic pain induced by chemotherapeutic drugs. The Rb fraction was continued after the paclitaxel injection was terminated and the thermosensitive response of each group of animals was continued to be measured.

As a result: the Rb component has a remarkable inhibitory effect on rat peripheral nerve pain caused by paclitaxel. As shown in fig. 19, the hindpaw withdrawal latencies of the two groups of animals were comparable before paclitaxel administration, and were significantly shorter on days six, 8, 11, and 15 after paclitaxel administration than before paclitaxel administration, indicating that paclitaxel induced significant peripheral neuropathic pain (# # P <0.01, # # P < 0.001); the latency period was significantly longer in the Rb group at day 6 and later time points than in the model group (. P <0.01,. P < 0.001). The above experimental results further support the clinical use value of the Rb component in preventing and treating pain after cancer treatment, especially peripheral nerve pain caused by chemotherapeutic drugs.

Discussion and summary (examples 11, 12) -the Rb component protects against mitochondrial damage in different pathological mechanisms and exerts a corresponding pharmacological effect

Rotenone is an inhibitor of mitochondrial oxidative phosphorylation, and by reducing ATP production leads to pathological events that lead to oxidative stress injury, excitotoxicity and inflammatory response in the central nervous system, ultimately leading to nerve injury and cerebrovascular injury, leading to various pathological conditions, including impaired motor system function and cognitive function. Mitochondrial damage is a common pathological mechanism of neurotoxicity caused by paclitaxel and other chemotherapeutic agents, while peripheral neuralgia is a symptomatic manifestation of its toxic side effects. The effects of the Rb component on resisting neuroinflammation, cerebrovascular injury and nerve injury caused by rotenone and peripheral neuralgia caused by paclitaxel are fully proved, and the Rb component has protective effect on mitochondrial injuries caused by different causes, so that the Rb component is further supported to prevent and treat peripheral and central diseases taking mitochondrial function damage as a common pathological mechanism.

Claims

1. Application of panaxadiol saponin Rb component in preparing medicine for preventing and treating metabolic disorder related diseases, wherein the panaxadiol saponin Rb component mainly comprises panaxadiol saponin Rb₁、Rb₂、Rb₃The five panaxadiol saponins of Rc and Rd have the structural formula:

ginsenoside Rb₁(ginsenoside Rb₁):R＝–D–glucopyranosyl

Ginsenoside Rb₂(ginsenoside Rb₂):R＝–L–arabinopyranosyl

Ginsenoside Rb₃(ginsenoside Rb₃):R＝–D–xylopyranosyl

Ginsenoside Rc (ginsenoside Rc) R (L-arbinofurosyl)

Ginsenoside Rd (ginsenoside Rd), R ═ H,

it is characterized in that the preparation method is characterized in that,

the medicine is various medicinal preparations prepared by taking the panaxadiol saponin Rb component as a single active ingredient or being combined with other active compounds and pharmaceutically acceptable carriers, wherein the other active compounds comprise substances for reducing blood sugar level, traditional Chinese medicine active substances, natural products, artificially synthesized compounds and in-vivo active substances.

2. The use of a panaxadiol saponin Rb component of claim 1 in the manufacture of a medicament for the prevention and treatment of a disease associated with a metabolic disorder, wherein the disease associated with a metabolic disorder comprises neurodegenerative and neurological disorders and the peripheral and central neuropathic symptoms associated with cancer therapy.

3. The use of a panaxadiol saponin Rb component according to claim 1 in the preparation of a medicament for the prevention or treatment of a disease associated with metabolic disorders, wherein the medicament is prepared with one or two or more of the five panaxadiol saponin components as an active ingredient.

4. The use of the Rb component of ginsenoside Rb as in claim 1 or 3 in the preparation of a medicament for the prevention and treatment of metabolic disorder related diseases, wherein the total content of the five ginsenoside Rb is 50-98% of the Rb component, Rb₁、Rb₂、Rb₃The content of the five monomer compounds of Rc and Rd is 3-50% of the content of the Rb component, but the content of the five monomer compounds is more than 20% at the same time.

5. The use of a panaxadiol saponin Rb component according to claim 1 or 3 in the manufacture of a medicament for the prevention and treatment of a disease associated with metabolic disorders, wherein Rb is₁、Rb₂、Rb₃The content of the five monomer compounds of Rc and Rd respectively accounts for more than or equal to 10-30% of the content of the Rb component.

6. The use of a panaxadiol saponin Rb component of claim 2 in the preparation of a medicament for the prevention or treatment of a disease associated with a metabolic disorder, wherein the neurodegenerative disease is selected from the group consisting of alzheimer's disease, vascular dementia, parkinson's disease, multiple sclerosis and huntington's disease.

7. The use of a panaxadiol saponin Rb component of claim 2 in the manufacture of a medicament for the prevention and treatment of a disease associated with a metabolic disorder, wherein the neurological disorder comprises epilepsy, schizophrenia, depression.

8. The use of a panaxadiol saponin Rb component of claim 1 in the preparation of a medicament for the prevention and treatment of a disease associated with a metabolic disorder, wherein the medicament is formulated in the form of: oral solid or liquid preparations, sustained release preparations, controlled release preparations, targeted preparations, enteric preparations, or injection preparations, or effervescent preparations, or in ointments or creams for topical administration, suppositories, or spray preparations.

9. The use according to claim 8, wherein the medicament is administered orally or by injection or by nasal spray or by external application to the skin or by anal administration.