TWI609691B - MEDICAL USES OF Ophiocordycepsformosana IN TYPE 1 DIABETES AND COMPLICATIONS THEREOF - Google Patents

Abstract

The invention relates to Taiwan Cordyceps sinensis, in particular to the Taiwan Cordyceps sinensis isolate disclosed in the present invention, for the medical use in treating type 1 diabetes and depression. The present invention is also directed to Taiwan Cordyceps sinensis, in particular, the Taiwan Cordyceps sinensis isolate disclosed in the present invention, for use in promoting the adiponectin content in the body.

Description

Medical use of Taiwan Cordyceps sinensis in type 1 diabetes and its complications

The invention relates to the medical use of Taiwan Cordyceps sinensis, in particular to the medical use of Taiwan Cordyceps sinensis in the treatment of type 1 diabetes and depression. The invention further relates to the medical use of Taiwan Cordyceps sinensis for enhancing the adiponectin content in the body.

Generalized Cordyceps (Cordyceps s.l. (Sensu lato) ) entomopathogenic (entomopathogenic) fungi in some pharmaceutically active species, comprising Cordyceps sinensis (Ophiocordyceps sinensis) and pupae Cordyceps (Cordyceps militaris) and other species, but a very The traditional Chinese medicine, which has been famous for a long time, has been used as a tonic and folk prescription for many years, and it is also called "three treasures of traditional Chinese medicine" with ginseng and antler. Taiwanese Cordyceps sinensis ( Ophiocordyceps formosana ) was first discovered in 1981 by Japanese Kobayashi and Shimizu in the Xitou area of Nantou County, Taiwan (Kobayasi, Y., Bull. Natn. Sci. Mus., Tokyo. Ser. B 7, p. 113-122 (1981)), which was originally identified as a native species of Taiwan. AD 2001, Anhui University professor Li as Huangshan Mountain in Anhui Province to Taiwan also collected Cordyceps sinensis, Cordyceps distribution means is not limited to Taiwan Taiwan (Li, et al., Mycosystema 24, p.349-355 (2005)). Taiwan Cordyceps sinensis is very difficult to collect, so there are few reports in the literature, and there are not enough materials for further identification and discussion of its possible medical uses.

In the Republic of China Patent Application No. 104130492, the applicant of the present invention revealed a novel isolate of Taiwan Cordyceps sinensis, which has been shown to be suitable for the species of Taiwan Cordyceps sinensis by morphological, biochemical and phylogenetic analysis. A typical strain. The patent application further discloses that Taiwan Cordyceps sinensis is superior to other species in the general Cordyceps genus in terms of antioxidant activity and cancer cell inhibitory activity, and has lower toxicity to normal cells.

Diabetes is a metabolic disease characterized by high blood glucose levels above the standard for a long time. According to the World Health Organization, there were 382 million diabetic patients worldwide in 2013, and the number of patients worldwide is expected to grow to 592 million in 2035. In the United States, up to 29 million people suffered from diabetes in 2014, while in Taiwan it is estimated to be more than one million. In 2012, diabetes was still the fourth highest cause of death among residents in Taiwan. Diabetes not only seriously affects people's health, but also imposes a huge burden on the medical and financial systems of various countries.

Clinically, diabetes can be roughly divided into four types: type 1 diabetes, type 2 diabetes, gestational diabetes, and recurrent diabetes. Among them, type 1 diabetes is also called insulin-dependent diabetes. (IDDM), the cause may be due to chronic autoimmune disorders, viral infections or environmental factors and genetic factors, resulting in destruction of insulin-producing islet beta cells, causing the patient to stop producing insulin. Patients usually have to inject exogenous insulin to metabolize glucose in the body, and other treatments such as pancreas transplantation and beta cell transplantation are clinically limited, so the treatment effect is limited. Type 1 diabetes is common in children and adolescents compared to other types of diabetes, so it was once called "juvenile-onset diabetes." In Taiwan, according to the 2002 and 2004 surveys, type 1 diabetes accounts for approximately 2-4% of all diabetic patients.

Thiazolidinediones (TZDs) such as rosiglitazone are commonly used clinically to control blood sugar in diabetic patients, but the administration of such drugs often causes adverse side effects such as stomach pain, diarrhea, heart failure and edema.

In addition, diabetic patients not only face typical clinical problems such as weight loss, increased fluid consumption, frequent urination, etc. According to statistics, about 15% of diabetic patients have the risk of depression and anxiety, while type 1 diabetes patients Compared with patients with type 2 diabetes, they have higher scores on the Beck depression inventory score and thus have a higher risk of depression. Drugs for treating depression can be roughly classified into first-generation antidepressants and second-generation antidepressants. The first generation of tricyclic and tetra/heterocyclic antidepressants (TCAs) can cause hyperglycemia in humans. Although the second generation of selective serotonin re-uptake inhibitors (SSRIs) has been widely used as the first line of treatment for depression, these drugs are prone to nausea, anorexia, and incompetence. Such side effects are more likely to cause blood sugar disorders in diabetic patients. Accordingly, there is a need in the related art for a new pharmaceutical composition that is therapeutically or prophylactically effective for Type 1 diabetes and/or depression, and that is highly safe for humans and animals.

Guo et al. reported that Ophiocordyceps sinensis and vanadium can synergistically improve the symptoms of diabetes and depression in rats (Guo JY et al ., Evid Based Complement Alternat Med , 2011; Article ID 450316), but alone Tibetan Cordyceps has no effect. The Republic of China Patent Publication No. 201136599 discloses that a fermentation culture of Cordyceps sobolifera is useful for treating or preventing diabetes and its complications. Only a few species of the general Cordyceps species have been shown to be pharmaceutically active, and the indications for these medicinal Cordyceps species are also different. Based on the above literature, the pharmaceutical activity of Taiwan Cordyceps sinensis was not inferred.

The inventors of the present invention have conducted extensive research on the pharmaceutical activity of Cordyceps sinensis in Taiwan. Now, the inventor of the case found that Taiwan Cordyceps sinensis, especially the Taiwan Cordyceps isolate isolated in this case, can improve type 1 diabetes and its complications, and can also promote adiponectin and various neurotransmitters in adipose tissue. The content in the marginal zone of the brain. The inventors of the present invention discovered that the Japanese Cordyceps sinensis, especially the Taiwan Cordyceps sinensis isolate disclosed in this case, has a superior effect in improving anxiety and depression through the model of depression caused by the initiation of type 1 diabetes in mice. . The disclosure of this case shows that Taiwan Cordyceps sinensis is effective in the treatment of type 1 diabetes, its complications and depression, and thus can be used as a supplement or alternative medicine for the treatment of these diseases.

In accordance with a first aspect of the present invention, the present invention relates to the use of a Taiwanese Cordyceps sinensis for use in the manufacture of a medicament for treating Type 1 diabetes and its complications in a body. Preferably, the complication of the first type of diabetes is selected from the group consisting of hyperglycemia, diabetic ketoacidosis, hyperglycemia, hyperosmolar state, hypoglycemia, depression, weight loss, neuropathy, renal disease, inflammation, A group of cardiovascular and high cholesterol complications, ocular lesions, and liver lesions. More preferably, the complication of the first type of diabetes is selected from the group consisting of hyperglycemia, depression, and weight loss.

In accordance with a second aspect of the present invention, the invention relates to the use of a Taiwanese Cordyceps sinensis for use in the manufacture of a medicament for treating depression in a body.

In accordance with a third aspect of the present invention, the invention relates to the use of a Taiwanese Cordyceps sinensis for use in the manufacture of a medicament for increasing adiponectin content in a body.

Unless otherwise stated, the following terms used in the patent specification and claims of this case have the definitions given below. Please note that the singular expression "a" used in the description and the claims is intended to cover one or more items, such as at least one, at least two, or at least three, A matter of inclusion. In addition, the open connection words such as "including" and "having" used in the claims of the present invention are combinations of elements or components described in the claims, and do not exclude other elements or components not specified in the claims.

The term " Tianjin Cordyceps sinensis" as used in this specification is intended to cover all isolates classified as Cordyceps formonsana and classified in Ophiocordyceps formosana , in particular based on the nucleotide sequences of the RPB1 , RPB2 and EF1-alpha genes. Identity , and is classified as all isolates of this species in Ophiocordyceps formosana . These strains of Cordyceps sinensis in Taiwan include strains of Cordyceps sinensis that are readily available to those of ordinary skill in the art, such as isolates that can be purchased from domestic or foreign depository institutions or obtained through application for distribution, or use in the art. A strain of Taiwan Cordyceps sinensis isolated from a natural source by a conventional microbial separation method. In a preferred embodiment, the Taiwan Cordyceps sinensis is the Taiwan Cordyceps sinensis isolate disclosed in the present case, which is deposited under the registration number of BCRC 930172 in the Food Industry Research and Development Institute (FIDI). The Biosource Collection and Research Center (BCRC), and the DSM 32000 registration number are deposited with the German National Collection of Microorganisms (DSMZ).

The isolates of Taiwan Cordyceps sinensis disclosed in this case can be cultured and propagated according to the conventional Cordyceps mycelium culture method. In general, the culture conditions suitable for Cordyceps mycelium are glucose, sucrose, galactose, fructose, corn starch, malt extract or a combination thereof as a carbon source, and ammonium sulfate and ammonium nitrate are used. , sodium nitrate, casein amino acid, casamino acid, yeast extract, peptone and tryptone or a combination of them as a nitrogen source, and adjust the pH of the medium To neutral to slightly acidic, it is preferably pH 5.0-7.0, such as pH 5.5-6.0. In a preferred embodiment, the medium is a liquid medium suitable for shake flask fermentation to obtain Cordyceps mycelium, such as potato dextrose broth. In another preferred embodiment, the medium is a solid medium such as potato dextrose agar (PDA) or Sagittide dextrose agar extract (S-DAY). The culture temperature is preferably controlled at room temperature or slightly below room temperature, for example, 15 ^o C-25 ^o C.

In general, Cordyceps sinensis is not easy to produce fruiting bodies by artificial culture, and it is usually only possible to mass produce mycelium by fermentation technology. Although the fruiting body of Cordyceps sinensis has been successfully cultured by artificial culture, the fruiting body takes a long time to grow, and the fruiting rate of the fruiting body is low and the mutation rate is high, so the production of mycelium is still relatively high. Economically beneficial. As is known to those of ordinary skill in the art, the content of active ingredients such as adenosine and cordycepin in Cordyceps sinensis is much higher than that of mycelium, so it is generally recognized that Cordyceps sinensis is compared to mycelium. Has a higher medicinal value. However, according to the disclosure of the present specification, Taiwan Cordyceps sinensis, especially the Taiwan Cordyceps sinensis isolates disclosed in this case, can also exhibit superior medicinal activity.

The term "type 1 diabetes" as used in this specification generally refers to diabetes caused by the destruction of pancreatic beta cells, resulting in the pancreas being unable to secrete insulin or the secretion is too low. Most of the complications of type 1 diabetes are due to the imbalance of blood glucose levels, and patients with type 1 diabetes have a higher incidence of complications than patients with type 2 diabetes.

In particular, in patients with type 1 diabetes who are not regularly injected with insulin, hyperglycemia, the typical symptom of hyperglycemia, occurs. If type 1 diabetes patients continue to be in a state of low insulin, the body can not use glucose as a source of energy, and instead use the body's fat to replace, the excess ketone body produced by fat metabolism accumulates in the body and leads to diabetic ketone. Diabetic ketoacidosis (DKA), in the long run, will cause weight loss. Common symptoms of diabetic ketoacidosis include: hunger, extreme thirst, frequent urination, difficulty breathing, abdominal pain, arrhythmia, hypotension, shock, exhalation with sour or fruity taste, anorexia or even nausea, vomiting, etc. In addition, hyperglycemia in the body can also cause osmotic diuretic effects, resulting in a significant dehydration of water from a large loss of urine. If the water is not properly replenished, it will cause hyperglycemic hyperosmolar state (HHS). The patient's intentional state will gradually deteriorate, and the muscles of the extremities will twitch and cause coma and shock in severe cases.

In addition, since patients with type 1 diabetes require long-term injection of insulin to control blood sugar, once the insulin injection is excessive, the blood glucose concentration is too low, causing acute hypoglycemia in the clinic. Hypoglycemia often occurs with heartbeat, night sweats, pale skin, restlessness, numbness in the limbs, lethargy, confusion, headache, and other symptoms.

Diabetic patients usually combine a variety of risk factors that cause cardiovascular disease. Long-term imbalance of blood glucose concentration causes damage to large and small blood vessels. Hyperglycemia can cause arteriosclerosis, which also thickens the microvascular wall, thickens the blood, and reduces the skin. The blood circulation of the arms, legs and feet also changes the blood circulation of the eyes and the kidneys, causing chronic and irreversible complications. Macrovascular disease refers to the coronary artery or cerebrovascular, while small vessel disease refers to micro-arterial lesions such as the retina, kidney and peripheral neuropathy. Neuropathy caused by diabetes can cause paralysis, sometimes causing pain and weakness in the hands, arms, feet, and legs. Neuropathy can also cause problems in the digestive system, heart and sexual organs. If the injury occurs in the small arteries of the lower extremities, it will cause diabetic foot, which is the main cause of amputation of diabetics.

Type 1 diabetes is almost impossible to cure. Once it is not detected early and properly treated, it is more likely to cause irreversible chronic complications, which may lead to multiple concurrent stresses due to prolonged deterioration of the condition, changes in lifestyle and dietary expenditures, and increased medical expenditures. Anxiety disorders and depression. The symptoms of depression, depression, anxiety, nervousness and anger caused by stress in the patient are more likely to promote the secretion of cortisol, adrenaline and glycemic hormone in the body, making the symptoms of hyperglycemia more obvious and causing the deterioration of diabetes.

The IUPAC designation of streptozotocin (abbreviated as STZ) is 2-deoxy-2-({[methyl(nitroso)amino)carbonyl]amino)-β-D-glucopyranose, It is a natural substance that is selectively cytotoxic to pancreatic beta cells. Therefore, it is often used in the related art to establish an animal model of type 1 diabetes. The present invention uses this conventional animal model to test the medical use of Taiwan Cordyceps in type 1 diabetes and its complications. According to the present invention, the administration of Cordyceps sinensis to Taiwan can improve the typical symptoms of type 1 diabetes, namely hyperglycemia. As mentioned above, the imbalance of blood glucose levels caused by hyperglycemia is the fundamental factor for the subsequent acute and chronic complications of type 1 diabetes, so the administration of Cordyceps sinensis in Taiwan will also help to improve these complications, and the examples in this case are also illustrated. It has been shown that the administration of Cordyceps sinensis in Taiwan can indeed improve the complications such as weight loss, anxiety and depression caused by type 1 diabetes.

Accordingly, one aspect of the present invention is the use of a Cordyceps sinensis for the manufacture of a medicament for treating Type 1 diabetes and its complications in a single body. As used herein, "complication" means acute and chronic symptoms that manifest as a result of the development of type 1 diabetes and its course of treatment. Preferably, the complications of the first type of diabetes are selected from acute complications such as hyperglycemia, diabetic ketoacidosis, hyperglycemia, hyperosmolar state, and hypoglycemia, as well as depression, weight loss, and neuropathy ( Including but not limited to acral paralysis, bladder weakness, constipation, diarrhea, sexual incompetence and loss of consciousness), renal lesions (including but not limited to glomerulonephritis, renal glomerulosclerosis, renal syndrome, hypertensive nephrosclerosis) , uric acidemia, end stage renal disease and uremia), inflammation, cardiovascular and high cholesterol complications (including but not limited to stroke, myocardial infarction, coronary thrombosis, angina pectoris, heart failure, poor peripheral circulation, gangrene and foot infections) ), a group of chronic complications of ocular lesions (including but not limited to retinopathy, cataracts, glaucoma, and visual field reduction) and liver lesions including, but not limited to, liver fibrosis, fatty liver, and cirrhosis. More preferably, the complication of the first type of diabetes is selected from the group consisting of hyperglycemia, depression, and weight loss.

In one embodiment, the complication of the first type of diabetes is hyperglycemia, which means a blood glucose value that is higher than a reference fasting blood glucose level of a normal individual. For human individuals, blood glucose values above 100 mg/dl are usually given, such as a fasting blood glucose level above 110 mg/dl, or a blood glucose level higher than 140 mg/dl after 2 hours of eating. As shown in Example 3 below, hyperglycemia caused by type 1 diabetes can be improved by administering Cordyceps sinensis to Taiwan.

In one embodiment, the complication of the first type of diabetes is weight loss, which refers to any reduction in body weight, body mass index (BMI), or other type of body mass metric in a type 1 diabetic patient. As shown in Examples 2 and 3 below, both weight loss and subcutaneous adipose tissue reduction induced by Type 1 diabetes can be improved by administering Cordyceps sinensis to Taiwan.

In a specific example, the complication of the first type of diabetes is depression, which refers to a depression or loss of interest that occurs for at least two weeks without medical treatment, and the scope is intended to cover a general mood from a poor mood. All clinical syndromes of mild depressive disorder and even major depressive disorder, preferably the fourth edition of The Diagnostic and Statistical Manual of Mental Disorders, published by the American Psychiatric Association or The method contained in the fifth edition (referred to as DSM-IV and DSM-5) was diagnosed and determined to be depression. Since anxiety and depression overlap in clinical symptoms, and many antidepressants also have anti-anxiety effects on medication, the "depression" described in this application also covers anxiety disorders. In a preferred embodiment, the "depression" includes, but is not limited to, severe depression, minor depression, bipolar disorder, persistent dysthymia, persistent palsy. Cyclothymia, pre-menstrual syndrome, premenstrual dysphoric disorder, generalized anxiety disorder, panic disorder (phobia, including social phobia) Specific subjects such as claustrophobia and agoraphobia, panic disorder, panic disorder, obsessive compulsive disorder, and post-traumatic stress disorder, overeating Bulmiia, anorexia, adolescent depression, and trichotillomania.

This application uses three widely used animal behavior tests for depression and anxiety to evaluate the therapeutic effect of Taiwan Cordyceps on depression.

The elevated plus maze test (EPM) was first proposed by Hendry et al. in 1984. The principle is that rodents, such as mice, have an inferred nature for the unknown new environment, but are in the open arms of the elevated cross maze. It also creates fear because of its aversion to high and empty nature, which creates ambivalence to simulate anxiety symptoms. Because the degree of preference of the mouse for the open arm is negatively correlated with the degree of anxiety, it means that the longer and more frequent the mouse enters the open arm, the lower the degree of anxiety.

The tail suspension test (TST) is to suspend the mouse tail and hang it in the air in an upside down state, calculate the time it takes to wriggle and not wriggle. The longer a mouse becomes helpless and no longer breaks away from writhing, the higher the degree of disappointment and self-abandonment.

The principle of the open field mission test (OFT) is that the edge of the test device is walled, rodents such as mice tend to be relatively safe in the surrounding activities, and the central open space is the wilderness without the shackles. If the number and time of mice staying in the open area increases, it means that the degree of anxiety is reduced.

As shown in Example 5 below, the above three animal behavior tests consistently showed that the symptoms of depression caused by type 1 diabetes can be improved by administering Cordyceps sinensis to Taiwan. However, according to another aspect of the present invention, type 1 diabetes is selected as a representative mode of depression, because the typical symptoms of depression caused by it are also common in many other types of physiology and psychology. And patients with environmental stress. Accordingly, the present application also encompasses the use of a Taiwanese Cordyceps sinensis for the manufacture of a medicament for treating depression in a body, wherein the "depression" is not limited to being caused by type 1 diabetes. It also covers depression caused by other chronic diseases, drug abuse, alcohol abuse, bereavement or other environmental stresses.

The application further encompasses the use of a Taiwanese Cordyceps sinensis for the manufacture of a medicament for increasing the adiponectin content in a body. Adiponectin, also known as GBP-28, apM1, AdipoQ and Acrp30, is a protein hormone secreted by adipose tissue into the blood, which acts on the metabolism of glucose and the oxidation of fatty acids. It is known that the low concentration of adiponectin in the blood is associated with the development of insulin resistance, hypertension, high triglycerides, and atherosclerosis. Compared with people who do not have diabetes, the adiponectin content of diabetic patients is usually low, and adiponectin helps to improve the body's sensitivity to insulin (Yamauchi T. et al ., Nat Med . 2001 Aug; 7(8): 941-6). Increasing the concentration of adiponectin in the blood or brain can improve the melancholic performance of experimental animals (Liu, J., et al ., Proc Nat Acad Sci, 2012 , 109: 12248-12253; and Yau SY et al ., Proc Nat Acad Sci, 111: 15810-15815). These facts point out that increasing adiponectin levels in the body can not only improve diabetes and its complications, but also help to improve the symptoms of depression.

As shown in Example 4 below, the Taiwan Cordyceps extract can up-regulate the amount of endogenous adiponectin in the SAT and the concentration in the blood circulation. In the case of a human individual, the term "adiponectin" refers to adiponectin precursor (SEQ ID NO: No. 1), deletion of 14 or 17 amino acid-directed sequences at the N-terminus of the precursor. Mature full length adiponectin, their post-translational modifications (including glycosylated forms and globular adiponectin), and their natural oligomers such as trimers, hexamers and Dimer. The term "promoting adiponectin content" refers to raising the level of endogenous adiponectin in the body after administration to a higher level than before administration, by taking individual blood samples and measuring the lipids therein. The content of the combination is used for judgment. In the case of humans, the amount of adiponectin in the blood varies depending on the individual's constitution and gender, and is generally between 5 and 10 μg/ml. In the case where the individual giving Taiwan Cordyceps sinensis is human, it is preferred that the adiponectin level in the blood is increased to a concentration of 5 to 10 μg/ml or higher, for example, a concentration of 1 μg/ml to 100 μg/ml. Preferably, the concentration is from 5 micrograms per milliliter to 75 micrograms per milliliter, for example from 10 micrograms per milliliter to 50 micrograms per milliliter.

As is known to those of ordinary skill in the related art, the occurrence of depression is closely related to the lack and imbalance of monoamine neurotransmitters such as dopamine, serotonin and norepinephrine in the brain. Therefore, many anti-depression and anti-anxiety drugs are currently aimed at improving monoamine neurotransmitters in the marginal zone of the brain. It is worth noting that the serotonin content in the hypothalamus increases with hyperglycemia caused by diabetes, meaning that controlling blood glucose levels also helps to improve depression. The data in Example 6 below shows that the extract of Cordyceps sinensis in Taiwan can significantly increase the content of the above-mentioned neurotransmitters and their metabolites in the marginal zone of the brain.

Although not wishing to be bound by any theory, the disclosure of this application shows that Taiwan Cordyceps may improve symptoms of depression through at least three different ways: (1) lowering blood glucose levels; (2) improving adiponectin performance in the body And (3) increase the content of monoamine neurotransmitters in the marginal zone of the brain.

The terms "treatment" and the like as used herein mean reversing, alleviating, delaying, or inhibiting the progression of the disease described in the present specification or one or more of its symptoms. In some embodiments, the treatment can be performed after developing one or more symptoms. In other embodiments, the treatment can be performed asymptomatically. For example, treatment can be performed to an susceptible individual prior to the onset of symptoms (eg, based on the history of symptoms and/or based on genetic or other susceptibility factors). Treatment can also be continued after the symptoms have been resolved to prevent or delay the recurrence.

According to the disclosure of the present invention, any amount of Taiwan Cordyceps sinensis which can effectively treat the diseases described in the present specification or which can effectively alleviate the severity of these diseases can be administered to a subject in need through any administration route. The exact amount required will vary from individual to individual, depending on the species, age and health of the individual, gender and diet, severity of illness, specific dosage form, mode of administration, duration of treatment, and agents used in combination or concurrently. Depending on factors. Those with ordinary knowledge in the related art should be able to understand that the daily total dose of Cordyceps sinensis in Taiwan will be determined by the physician within the scope of sound medical judgment. When administered to a human individual, the extract of Cordyceps sinensis in Taiwan is usually administered at a frequency of about 0.01 to 1000 mg/kg body weight per one to three times a day, once to three times a week or once every two weeks, preferably. It is administered in an amount of about 0.1 to 100 mg/kg body weight per time, more preferably in an amount of about 1 to 50 mg/kg body weight per time. In a specific example, the amount of Cordyceps sinensis in Taiwan can be such that the blood glucose level of the human individual is lower than the blood glucose level of about 100 mg/dl, for example, the blood sugar level is less than about 110 mg/dl, or the blood sugar after eating for 2 hours. The value is less than about 140 mg/dl. In another specific example, the amount of Cordyceps sinensis in Taiwan can be such that the adiponectin content in the blood of the human individual reaches a concentration of from 1 μg/ml to 100 μg/ml, preferably from 5 μg/ml to 75 μg/ml. For example, a concentration of 10 μg/ml to 50 μg/ml.

The term "individual" as used in this description is intended to encompass human or non-human vertebrates, such as non-human mammals. Non-human mammals include livestock animals, companion animals, laboratory animals, and non-human primates. Non-human mammals also include, without limitation, horses, cows, pigs, goats, dogs, cats, mice, rats, guinea pigs, hamsters, rabbits, baboons, and the like. It is preferably human, and more preferably a human subject suffering from type 1 diabetes and its complications, or a human subject suffering from depression.

For the purposes of research, the term "individual" as used in this specification is also intended to encompass cells, tissues and organs. The invention disclosed in the present invention is thus applicable not only to in vivo but also to in vitro.

According to the present invention, Taiwan Cordyceps sinensis can be administered to an individual in the form of fruit bodies and mycelia. Since the fruit body is not easily cultured, it is preferably administered to the individual in the form of a mycelium, more preferably in the form of a dried pulverized particle of a mycelium, an aqueous extract or a preparation of an aqueous extract.

The dried Cordyceps sinensis fruit body or mycelium can be directly ground using a grinder to prepare the aforementioned dry pulverized particles. The particle size of the dried pulverized particles is not particularly limited as long as it is suitable for direct administration to an individual and is advantageous for the intake and absorption of the individual, or is advantageous for further processing into other forms suitable for administration to an individual.

Any conventional process can be used to prepare the aqueous extract of Cordyceps, as long as the extraction process can effectively transfer the water-soluble and water-miscible components of Cordyceps from the solid portion of Cordyceps to an aqueous solvent. Generally, the extraction process comprises pulverizing the fruiting body or mycelium of Cordyceps sinensis, and then lowering the alcohol (for example, methanol) having 1 to 6 carbon atoms and preferably 1 to 4 carbon atoms. An aqueous solvent such as ethanol or a combination thereof is used for the extraction operation, and it is preferred to apply heat, ultrasonic vibration or mechanical agitation at the same time to facilitate the extraction efficiency, and finally by filtration, centrifugation, etc. The solid portion is removed to obtain an aqueous extract of Cordyceps. Preferably, the extraction process is selected from the group consisting of a leaching method, a hot water reflux method, a hot alcohol reflux method, an ultrasonic water extraction method, and an ultrasonic alcohol extraction method. In a preferred embodiment, the aqueous extract of Taiwan Cordyceps sinensis is prepared by hot water reflux method, that is, the quantitative hot water is continuously flowed through the cordyceps material to extract the aqueous extract, preferably the hot water temperature is 30-100 ^o C, more preferably 40-60 ^o C, most preferably 45-55 ^o C, e.g. 50 ^oC, and the extraction operation for 30 minutes to 12 hours, preferably 1-6 hours, for example 2 hours. In a preferred embodiment, the efficiency of the extraction operation can be assessed by the amount of extract of Cordyceps active ingredients such as D-mannitol, adenosine and/or cordycepin.

The aqueous extract of the aforementioned Taiwan Cordyceps sinensis can be formulated into an appropriate concentration and then formulated into an oral form or a parenteral form for administration to an individual. When these administration forms are prepared, a diluent such as a filler, a bulking agent, a binder, a wetting agent, a disintegrating agent, a surfactant, or the like may be added, or other types of excipients may be added.

The form of the liquid preparation for oral administration includes a suspension, a solution (syrup, a drink, etc.), an emulsion, and the like. In addition to the most commonly used diluents such as water or liquid paraffin, wetting agents, sweeteners, flavoring agents and preservatives may be added. The form of the solid preparation for oral administration includes tablets, pills, powders, granules, capsules, lozenges and the like.

Forms for parenteral administration include sterile solutions, non-aqueous solutions, suspensions, emulsions, lyophilizates, and the like. Solvents for use in solvents, suspensions or emulsions for non-aqueous solutions include, but are not limited to, vegetable oils, propylene glycol, polyethylene glycol, olive oil, and ethyl oleate.

In general, these preparations are prepared by adding at least one excipient to the pulverized granules or aqueous extract, if necessary, and then granulating, tableting or encapsulating. The excipients include, but are not limited to, water, lower alcohols having from 1 to 4 carbon atoms, natural or synthetic oils, gelling agents, suspending agents, emulsifiers, thickeners, inert powders, natural and synthetic Polymers, humectants, sweeteners, flavoring agents, fragrances, perfumes, colorants, preservatives, lubricants, surfactants. Preferred excipients include, but are not limited to, water, ethanol, starch, calcium carbonate, sucrose, lactose, gelatin, gums, dextrin, cyclodextrin, microcrystalline cellulose, carboxymethyl cellulose, cross-linked carboxy Methyl cellulose sodium and the like. Further, a lubricant such as magnesium stearate or talc may be added in addition to the excipient alone.

The Cordyceps sinensis composition disclosed in the present invention comprises an aqueous extract of the Cordyceps sinensis fruit or mycelium of Taiwan or a dry pulverized granule, and optionally contains an acceptable excipient. The term "acceptable excipient" as used in this context means an inert substance which is used as a carrier for the composition of Cordyceps sinensis in this case, which is not toxic, irritating, pyrogenic, antigenic and hemolytic to the individual to be administered, and No substantial pharmacological activity will not hinder the beneficial effects of the composition of Cordyceps sinensis in this case. The composition of Cordyceps sinensis disclosed in this case can be directly eaten, eaten, or blended into various edible materials such as beverages, snacks, animal feed, dietary supplements, other Chinese herbal medicines, or packaged into other suitable forms. For example, the aforementioned dry pulverized granules are packaged in a tea bag in the form of a tea bag for brewing. In a preferred embodiment, the composition of Cordyceps sinensis disclosed in the present invention may be combined with a composition derived from other Cordyceps species, for example, with a fruiting body or mycelium composition from Cordyceps sinensis and/or Cordyceps sinensis, thereby being formulated into A health food and pharmaceutical that is suitable for individual intake and that combines safety and efficacy.

In another preferred embodiment, the Cordyceps composition of the present invention consists essentially of an aqueous extract of the Taiwan Cordyceps sinensis isolate as described above or a dry comminuted granule and the aforementioned acceptable excipient. The term "consisting essentially of" as used herein means that the other combinations of the components described above do not exclude other undescribed components which do not substantially affect the properties of the aqueous extract or the dried pulverized particles. In another preferred embodiment, the Cordyceps sinensis composition of the present invention consists solely of the aqueous extract of the aforementioned Cordyceps sinensis isolate or the dried comminuted granules and the aforementioned acceptable excipients.

The following examples are for illustrative purposes only and are not intended to limit the scope of the invention. All data shown in the examples was done via at least three independent experiments and is expressed as mean ± standard deviation. Statistical analysis was performed using a single factor analysis of variance (ANOVA) followed by a Tukey's test or a Student's test, and its significance was determined by the manner described in the examples.

Example 1 : Preparation of Taiwan Cordyceps Extract ( OFE ) The extract of Taiwan Cordyceps sinensis used in the examples in this case was deposited with the registration number of BCRC 930172 at the Food Industry Research and Development Institute. , FIRDI) Biosource Collection and Research Center (BCRC), and the Japanese Cordyceps sinensis isolates deposited with the National Collection of Microorganisms and Cultures (DSMZ) under the DSM 32000 registration number, using the Republic of China patent The method disclosed in the application No. 104130492 is produced. Briefly, the mycelium of the above-mentioned Taiwan Cordyceps isolate was transferred to potato dextrose agar medium containing 0.4% potato starch, 2% dextrose and 1.5% agar (PDA; purchased from Maryland, USA). Difco Becton Dickinson, Sparks), and cultivated at 25 °C for 28 days. The colonies received were dried at 55 ° C until the next day and stored in a desiccator for subsequent analysis or production of the extract.

The dried Cordyceps sample was pulverized at a running speed of 1300 rpm using a homogenizer (SH-100 homogenizer from Kurabo, Japan). In a 50 ml Falcon® centrifuge tube (purchased from Becton Dickinson and Company, USA), 1 gram of Cordyceps sinensis powder was dissolved in deionized water at a ratio of solid to solvent of 1:40 and subjected to shaking 30 second. Next, the hot water was refluxed at 50 ° C for 2 hours to carry out an extraction operation (an ultrasonic wave was applied for the first 30 minutes). After centrifugation at 4000 rpm (about 3200 x g) for 20 minutes, the supernatant was collected and filtered through a 0.22 micron pore size filter (Millipore, USA).

The obtained extract of Taiwan Cordyceps sinensis was subjected to high performance liquid chromatography (Hitachi D-2000 HPLC management system) to monitor the quality and content of the contents in the extract. High performance liquid chromatography was performed on a RP-18 column (150252 Purospher® STAR RP-18 endcapped (5 μm) LiChroCART® 250-4; Merck, USA) at a flow rate of 1 ml per minute. The composition of the mobile phase was 20% methanol/H ₂ O. Extracts were stored for use in the following Examples -20 ^oC.

Example 2 : Diabetic animal model 30 8-week-old male C57BL/6 mice (average body weight 20 ± 3 g) were purchased from the Experimental Animal Center (NLAC) of the National Experimental Research Institute of the City of Taipei. The temperature of the reared mice was maintained at 21 ± 3 ^o C, the relative humidity was 50 ± 10%, and the day and night cycle was 12/12 hours. Groups and trials were performed after the mice were acclimated for 7 days in standard animal rooms. After 24 hours of fasting for 24 mice, 40 mg/kg body weight of streptomycin (STZ; Sigma-Aldrich, St. Louis, Missouri, USA) was administered via intraperitoneal injection once daily for 5 consecutive times. On the day, the mice were allowed to rest for 2 days. After confirming that the 24 streptomycin-treated mice showed symptoms of diabetes, they were randomly divided into three groups, each of which was fed with 100 μl of phosphate-buffered physiological saline (STZ-PBS group), 25 mg of Example 1 The extract of Taiwan Cordyceps sinensis (STZ-OFE group) prepared in the middle, or 10 mg of rosiglitazone (Sigma-Aldrich, USA) (STZ-ROSI group) lasted for 28 days. The remaining 6 mice that did not receive streptomycin were given 100 μl of PBS as a control group. Mouse body weight and feeding conditions were recorded daily. Blood samples were taken before sacrifice in the mice and stored at -80 ^o C for blood analysis. After sacrificing the mice, islet cells were taken and counted. Subcutaneous adipose tissue (SAT) was collected, weighed, and the ratio of tissue to body weight was calculated. Brain tissues such as prefrontal cortex, striatum, and hippocampus located in the mouse brain were also quickly taken out and weighed. See Figure 1 for the experimental flow of each case in this case.

As shown in Fig. 2C, compared with the STZ-PBS group, the STZ-OFE group significantly accumulated more subcutaneous adipose tissue, even more than the subcutaneous adipose tissue of the STZ-ROSI group. In addition, in the diabetic mouse model induced by STZ, a decrease in the number of islet β cells can be observed, and neither the ROSI nor the OFE can restore the number of β cells during the experiment (data not shown), which means that The therapeutic effect of Taiwan Cordyceps sinensis on the complications of type 1 diabetes is not through an increase in the number of beta cells.

Example 3 : Intraperitoneal glucose tolerance test ( GTT ) is shown in Figure 1. Before and after the feeding treatment (represented by PRE GTT and POST GTT, respectively), the mice were fasted for 6 hours, and each mouse was administered by intraperitoneal injection. 2 g / kg body weight of glucose. At 0, 30, 60, and 120 minutes after glucose infusion, blood samples were taken from the tail of the mice, respectively, and blood glucose concentrations were measured using an Accu-Chek® active blood glucose meter (Roche Diagnostics, Inc., Mannheim, Germany).

As shown in Fig. 2A, a typical symptom of type 1 diabetes, that is, a decrease in body weight, is exhibited in the diabetic mouse model induced by STZ. Figure 2B further shows that in the diabetic mouse model induced by STZ, if no drug is administered, the fasting blood glucose level exceeds 200 mg/dL (392.2 ± 38.7 mg/dL in the STZ-PBS group). Relatively speaking, the fasting blood glucose of the control group was only 119.5 ± 24.2 mg/dL (p < 0.001). However, when the Taiwan Cordyceps extract or rosiglitazone was given, the above symptoms of diabetic mice were significantly relieved (STZ-OFE group and STZ-ROSI group). In particular, the area under the curve (AUC) of the fasting blood glucose level of the mice in the STZ-OFE group after receiving the glucose tolerance test (POST GTT) was significantly lower than that of the STZ-PBS group, almost equivalent to the control group. It shows that Cordyceps sinensis can not only improve the glucose metabolism of type 1 diabetes, but also promote the use of energy in patients, thus improving the complications of type 1 diabetes.

Example 4 : Immunohistochemical analysis The subcutaneous adipose tissue (SAT) collected in Example 2 was fixed with 4% paraformaldehyde and embedded in paraffin for sectioning. The tissue sectioned paraffin was then removed and treated at 95 ° C in an antigen retrieval solution (Abcam, Inc., Cambridge, MA; number ab93680) for 15 minutes to expose the antigen and equip with 80% methanol. The 0.3% H ₂ O ₂ solution in the solution was treated for 30 minutes to reduce the content of endogenous peroxidase. Sections were treated with 1% bovine serum albumin (BSA) and subsequently incubated with anti-mouse adiponectin antibody (Abcam, Inc., Kanqiao, MA; number ab22554) for 18 hours at 4 °C. Subsequently, sections were incubated with goat anti-mouse secondary antibody for 30 minutes and stained using EnVision® + Dual Link System HRP (DAB+) kit (Dako Cytomation, CA). The sections were over-stained with hematoxylin, rinsed with secondary deionized water for 10 minutes, and fixed with glycerin gel (Surgipath Medical Ind., USA). The slice images were taken, and the signal intensity of adiponectin was measured by selecting 5 sites in each image, and the image analysis software Image J (developed by the National Institute of Health, Bethesda, Maryland, USA) was used to calculate the small groups. The amount of adiponectin in the SAT of the mouse.

Figure 3A shows the adiponectin content in the plasma of each group of mice, wherein the three groups of mice given STZ had lower adiponectin in the blood than in the control group, however, in the STZ-OFE group. Plasma adiponectin was significantly higher than mice given PBS alone (STZ-PBS group). This data indicates that the administration of Cordyceps sinensis can restore part of the blood adiponectin content. The representative adiponectin-stained section image of SAT is magnified 40 times, as shown in Figure 3B, and the measured adiponectin signal intensity values in the image are shown in Figure 3C. Figures 3B and 3C indicate that the adiponectin re-synthesized in the SAT of the STZ-OFE group and the STZ-ROSI group was increased by 16% and 27%, respectively, compared to the STZ-PBS group. Taken together, these data show that the extract of Cordyceps sinensis in Taiwan can up-regulate the content of adiponectin in the SAT and blood, thereby promoting the biosynthesis of SAT.

Example 5 : Animal Behavioral Test This example was judged by measuring the performance of the mouse model established in Example 2 in the open field task test (OFT), the elevated plus maze test (EPM), and the tail suspension test (TST). The degree of anxiety and depression. A monitor is installed in a muffler room equipped with vents to monitor and record the complete test procedure. In OFT, mice were individually placed in the center of a field device (43.3 × 21.7 × 20.8 cm, made of transparent plexiglass) and allowed to freely explore for 60 minutes. The total moving distance of each mouse, the residence time in the central region, and the speed of movement in the central and peripheral regions were analyzed by TOP SCAN v2.0 software (Clever Sys, Reston, VA). In the EPM, an elevated cross maze with two open arms (30 cm long by 5 cm wide) and two closed arms was raised to a height of 50 cm from the floor. The closed arm has the same dimensions as the open arm but is otherwise enclosed by a 50 cm high transparent wall. Each arm body is made of a white acrylic plate and is arranged in a cross shape by a central platform (5 cm square). At the beginning of the experiment, each mouse was placed on a central platform and faced one of the closed arms. The movement path of each mouse within 5 minutes and the time spent in the device were recorded and analyzed by TOP SCAN v2.0 software (Clever Sys, Reston, VA). TST is used to measure the melancholy behavior of animals. In TST, 2-3 cm of the tail end of each mouse was adhered to the plastic tube on the ceiling of the muffler chamber by tape, so that the mouse was 10 cm away from the nearest object, 35 cm from the floor, and the tail distance was The ceiling is 38 cm and is soundproofed and light-shielded. The time during which each mouse was still in the experiment was calculated, and the behavior of each mouse in the test was scored by three testers. The term "stationary motion" as used herein is defined in a state in which the head and all of the claws of the mouse are completely stopped.

As shown in Figures 4A to 4C, the STZ-PBS group showed significant anxiety and depression behavior compared to the control group, indicating that STZ used to induce type 1 diabetes in mice and successfully established anxiety. Animal pattern of illness and depression. Figure 4A further shows that administration of ROSI or OFE can prolong the time that the mouse stays in the central region in the open field task test (OFT), where the ratio of residence time of the control mice to the total time in the central region is 0.18. ± 0.01%; the ratio of the STZ-PBS group was 0.12 ± 0.02%; the STZ-ROSI group was 0.23 ± 0.07%; and the STZ-OFE group was 0.24 ± 0.01%. In the elevated plus maze test (EPM), the STZ-OFE group had the longest residence time in the open arms, suggesting that the addition of Cordyceps sinensis extracts in Taiwan can promote the exploration behavior of mice. As shown in Fig. 4B, the ratio of residence time of the control mice in the open arms was 19.7 ± 9.3%; the ratio of the STZ-PBS group was 9.8 ± 4.7%; and that of the STZ-RODI group was 16.8 ± 7.8%; the STZ-OFE group It is 41.0 ± 12.3%). As also shown in Fig. 4C, in the tail suspension test (TST), the STZ-PBS group showed a significantly longer resting time than the control group, indicating that STZ caused symptoms of anxiety and depression. In contrast, the resting time of the STZ-OFE group of mice returned to only slightly higher than the control group. Based on the data obtained in this example, Taiwan Cordyceps can improve the pathological complications of anxiety and depression caused by type 1 diabetes.

Example 6 : Monoamine content in limbic regions In 1 ml of extract consisting of 0.1 M HCl, 10 ^-7 M ascorbic acid and 50 pg/μL isoproterenol, The brain tissues obtained in Example 2 were homogenized separately. The homogenate was then centrifuged three times at 13200 rpm for 4 minutes at 4 °C. The supernatant was filtered through a 0.22 micron pore size filter (Millipore, USA), and then injected into the reverse phase C18 column ^{(Hypersil TM GOLD aQ, 5μm,} 150mm × 4.60mm). By being equipped with an electrochemical sensor (produced by Shiseido Co., Ltd., Japan, model P/N 70-9143, range 10 nA, with 1 Hz filter and 0.7 V AppE battery) and equipped with an automatic sampler ( HPLC system of Dionex Corporation WPS-3000) to measure norepinephrine (NE), dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), 5-hydroxytryptamine (5-HT) and 5- The concentration of hydroxyindoleacetic acid (5-HIAA). The chromatographic mobile phase contained 0.17 M NaH ₂ PO ₄ , 0.63 mM ethylenediaminetetraacetic acid (EDTA), 0.60 mM octane-1-sulfonate sodium salt and 2.0 mM KCl, formulated in 20% methanol, and 85 % H ₃ PO ₄ adjusts the pH to pH 3.30. The flow rate was maintained at 0.6 ml per minute. Standards for the above compounds were purchased from Sigma-Aldrich, St. Louis, Missouri, USA. The lower limit of detection of the HPLC system for the sample is 0.1 pg.

The contents of neurotransmitters (DA and 5-HT) and their metabolites (DOPAC and 5-HIAA) in the marginal regions of the prefrontal cortex, striatum and hippocampus were shown in the table below. 1. Table 1. Neurotransmitter content in the marginal zone of the brain of each group of mice

As shown in Table 1, administration of STZ (STZ-PBS group) resulted in a decrease in the content of DA, DOPAC, 5-HT and 5-HIAA in the three marginal regions of the brain compared to the control group. However, mice given STZ received ROSI or OFE treatment to significantly restore the levels of these neurotransmitters and their metabolites in their prefrontal cortex and striatum. This data is consistent with the results of Example 5 and Figures 4A-4C showing that ROSI and OFE can improve anxiety and depression. Based on the data in this example, Taiwan Cordyceps sinensis can increase the content of neurotransmitters in the dopamine and serotonin signaling pathways, thereby improving the anxiety and depression symptoms caused by type 1 diabetes.

While the invention has been described in detail hereinabove, it will be apparent to those of ordinary skill in the art. The disclosure of the related art and the disclosure of the references discussed in the Detailed Description of the Invention and the Detailed Description of the Invention are hereby incorporated by reference.

no

Figure 1 is an overview of the experimental process of each example of the present case;

Figure 2A is a bar graph showing changes in body weight of each group of diabetic mice during the experiment;

Figure 2B is a bar graph showing the area under the curve (AUC) of the fasting blood glucose level obtained by the glucose tolerance test (POST GTT) of each group of diabetic mice after feeding;

Figure 2C is a schematic diagram showing the accumulation of subcutaneous fat tissue in each group of diabetic mice;

Figure 3A is a bar graph showing plasma adiponectin levels in each group of diabetic mice;

Figure 3B is an adiponectin stained slice image of white subcutaneous adipose tissue (SAT);

Figure 3C is a bar graph showing the adiponectin signal intensity values in the slice image of Figure 3B;

Figures 4A-4C are bar graphs showing the performance of each group of diabetic mice in the open field task test (OFT), elevated plus maze test (EPM), and tail suspension test (TST).

Domestic deposit information [please note according to the order of the depository, date, number] Republic of China Food Industry Development Institute, January 16, 104, BCRC 930172 Foreign deposit information [please note according to the country, organization, date, number order] German National Collection of Microorganisms (DSMZ), January 28, 104, DSM 32000

<110>Muqisheng Technology Co., Ltd. <120> Medical use of Taiwan Cordyceps sinensis in type 1 diabetes and its complications <160> Number of sequence identification numbers: 1 <210> Sequence identification number: No. 1 <211> Sequence Length: 244 <212> Sequence Type: PRT <213> Organism: Homo sapiens <400> Sequence: 1 Met Leu Leu Leu Gly Ala Val Leu Leu Leu Leu Ala Leu Pro Gly His 1 5 10 15 Asp Gln Glu Thry Thr Gry Gly Pro Gly Val Leu Leu Pro Leu Pro 20 25 30 Lys Gly Ala Cys Thr Gly Trp Met Ala Gly Ile Pro Gly His Pro Gly 35 40 45 His Asn Gly Ala Pro Gly Arg Asp Gly Arg Asp Gly Thr Pro Gly Glu 50 55 60 Lys Gly Glu Lys Gly Asp Pro Gly Leu Ile Gly Pro Lys Gly Asp Ile 65 70 75 80 Gly Glu Thr Gly Val Pro Gly A La Glu Gly Pro Arg Gly Phe Pro Gly 85 90 95 Ile Gln Gly Arg Lys Gly Glu Pro Gly Glu Gly Ala Tyr Val Tyr Arg 100 105 110 Ala Phe Ser Val Gly Leu Glu Thr Tyr Val Thr Ile Pro Asn Met 115 120 125 Pro Ile Arg Phe Thr Lys Ile Phe Tyr Asn Gln Gln Asn His Tyr Asp 130 135 140 Gly Ser Thr Gly Lys Phe His Cys Asn Ile Pro Gly Leu Tyr Tyr Phe 145 150 155 160 Ala Tyr His Ile Thr Val Tyr Met Lys Asp Val Lys Val Ser Leu Phe 165 170 175 Lys Lys Asp Lys Ala Met Leu Phe Thr Tyr Asp Gln Tyr Gln Glu Asn 180 185 190 Asn Val Asp Gln Ala Ser Gly Ser Val Leu Leu His Leu Glu Val Gly 195 200 205 Asp Gln Val Trp Leu Gln Val Tyr Gly Glu Gly Glu Arg Asn Gly Leu 210 215 220 Tyr Ala Asp Asn Asp Asn Asp Ser Thr Phe Thr Gly Phe Leu Leu Tyr 225 230 235 240 His Asp Thr Asn

Claims (10)

A use of Taiwanese Cordyceps sinensis ( Ophiocordyceps formosana ) for the manufacture of a medicament for treating Type 1 diabetes and its complications in a body. The use of claim 1, wherein the complication of the first type of diabetes is selected from the group consisting of hyperglycemia, diabetic ketoacidosis, hyperglycemia, high osmotic state, hypoglycemia, depression, weight loss, neuropathy, kidney A group of lesions, inflammation, cardiovascular and high cholesterol complications, ocular lesions, and liver lesions. The use of claim 2, wherein the complication of the first type of diabetes is selected from the group consisting of hyperglycemia, depression, and weight loss. A use of Ophiocordyceps formosana in Taiwan for the manufacture of a medicament for treating depression in a body. A use of Taiwanese Cordyceps sinensis ( Ophiocordyceps formosana ) for the manufacture of a medicament for enhancing adiponectin content in an individual having type 1 diabetes. The use of any one of claims 1 to 5, wherein the Taiwan Cordyceps sinensis is an aqueous extract of a Taiwan Cordyceps isolate, optionally comprising an acceptable excipient. The use of claim 6, wherein the aqueous extract of the Taiwan Cordyceps sinensis isolate is obtained by pulverizing the mycelium of Taiwan Cordyceps sinensis and refluxing the mycelium with hot water at a temperature of 40-60 ° C for a period of time. It was prepared by removing the solid portion from 30 minutes to 12 hours. For the use of claim 6, the Taiwan Cordyceps isolate is deposited with the Bioresource Conservation and Research Center (BCRC) of the Republic of China Food Industry Development Research Institute (FIRDI) under the registration number of BCRC 930172. The use of any one of claims 1 to 5, wherein the Cordyceps sinensis is a dry comminuted granule of a Taiwan Cordyceps isolate, optionally comprising an acceptable excipient. For the purpose of claim 9, the Taiwan Cordyceps isolate is deposited with the Biological Resources Preservation and Research Center (BCRC) of the Republic of China Food Industry Development Research Institute (FIRDI) under the registration number of BCRC 930172.