TWI446913B - A use of (22e,24r)-24-methyl-6β-methoxy-5α-cholesta-7,22-diene-3β,5-diol and 3β-hydroxy-(22e,24r)-ergosta-5,8,22-trien-7-one, natural active substances of cucurbita moschata - Google Patents

Description

Pumpkin active ingredient (22E, 24R)-24-methyl-6β-methoxy-5α-cholesterol-7,22-diene-3β,5-diol and (22E,24R)-3β-hydroxy ergot Sterol Use of -5,8,22-trien-7-one

The present invention relates to a plant extract and its use, and in particular to a pumpkin extract and its use for the preparation of a medicament for diabetes.

Diabetes is a chronic metabolic disease, mainly insulin deficiency or tissue loss of insulin response, leading to glucose metabolism dysfunction, resulting in hyperglycemia, and when the blood glucose level is too high, exceeding the renal tubular absorption limit, It will be excreted from the urine and cause diabetes. Among them, it can be divided into type 1 diabetes and type 2 diabetes according to the different conditions and causes of the disease.

Type 1 diabetes is insulin-dependent diabetes, and its cause may be that the body's autoimmune phenomenon destroys the pancreatic beta cells, resulting in insufficient insulin secretion. Current treatments for type 1 diabetes include insulin therapy, medial nutrition therapy (also known as diet planning and exercise planning), and administration of pramlintide, which controls blood sugar. )drug.

Type 2 diabetes is non-insulin-dependent diabetes mellitus, in which obese people are at high risk for type 2 diabetes. The reason may be that the fat cells of obese people continue to proliferate, leading to hypoxia of fat cells and promoting secretion of fat cells. A large number of inflammatory factors (such as TNF-α, IL-1, IL-6, etc.), and the above inflammatory factors stimulate insulin-sensitive tissues such as muscle, fat, liver cells, and produce insulin resistance. The drugs currently used to improve insulin resistance are quite limited, mainly thiazolidinedione (abbreviated as thiazolidinedione). TZD) drugs, such as pioglitazone, rosiglitazone, troglitazone, etc., these drugs are mainly regulated by the activation of the nuclear surface of the PPARγ receptor (peroxisome proliferator activated receptor gamma) The pathway of regulation, which regulates the function of intracellular glucose transporter gene transcription, promotes the increase of glucose uptake by cells, or activates AMPK of insulin-resistant cells, thereby promoting the displacement of GLUT4 from the cytoplasm to the cell membrane to improve the symptoms of type 2 diabetes. However, troglitazone has a severe hepatotoxicity report after administration and has been discontinued for clinical treatment.

In recent years, due to the prevalence of preventive medicine, the development of health foods that can be used to lower blood sugar has also received much attention. Such products include various plant ingredients such as bitter gourd, aphid, aloe, etc. However, bitter gourd has been shown to lower blood sugar, but The effective dose has not been confirmed so far; human sputum will interact with anti-depression, hypertension, estrogen and other drugs, and the body will produce hormone analogues during the treatment of human sputum, so blood pressure and mood must be monitored regularly when taking it. Aloe vera contains laxative ingredients, which can cause water and electrolyte imbalance in the body. Although it does not cause serious sequelae, it will eventually make users feel uncomfortable and difficult to obtain consistent recognition.

Since conventional medicines or health foods can cause discomfort to users, according to this, if the plant ingredients which have a significant effect on diabetes can be further found and applied to the development of therapeutic or preventive medicine, it is expected to be effective for all walks of life. Improve the health of Chinese people.

The squash is native to southern Asia, Africa and the Americas. It is an annual vine herb of the genus Cucurbitaceae. It is called the melon, papaya, squash, and winter melon. It is commonly known as the gourd in Taiwan. The pumpkin can be roughly divided into Chinese pumpkin ( Cucurbita moschata ), western pumpkin ( Cucurbita maxima (also known as Indian pumpkin), American pumpkin ( Cucurbita pepo ), black pumpkin ( Cucurbita ficifolia ), and Mexican pumpkin ( Cucurbita mixta ). The adaptability of pumpkin is very strong. It is widely cultivated in all parts of Taiwan and North and North. It is one of the main vegetables in summer and autumn. Conventional technology known that pumpkin can enhance the regeneration of liver cells, can improve pernicious anemia, and contains trace element zinc, can improve prostate hypertrophy, can treat frequent urination, urinary incontinence, impotence and premature ejaculation, but no pumpkin can be applied Controls related reports of blood sugar.

The main object of the present invention is to provide a pumpkin extract which contains a natural active ingredient which regulates blood sugar and which can improve diabetes.

A second object of the present invention is to provide a pumpkin extract which has a significant regulatory function for blood sugar of a diabetic patient and which does not cause side effects, and can be applied to the development of a drug for improving diabetes.

A second object of the present invention is to provide a pumpkin extract which is used as an active ingredient in the development of a medicament for diabetes patients.

In order to achieve the foregoing object, the technical means used in the present invention comprises: a pumpkin extract comprising: (22E, 24R)-24-methyl-6β-methoxy-5α-cholesterol-7,22-di Alkene-3β,5-diol; and (22E,24R)-3β-hydroxyergosterol-5,8,22-trien-7-one.

In the pumpkin extract of the present invention, the above pumpkin extract is obtained by extracting a pumpkin sample with methanol to obtain a pumpkin extract. Extracting the crude extract of the pumpkin with ethyl acetate and water to obtain an ethyl acetate layer and an aqueous layer; using a mixture of n-hexane and ethyl acetate as a mobile phase, the above-mentioned fixed on a rubber hose column The ethyl acetate layer was subjected to chromatographic separation, and the fraction extracted with a 70-80% ethyl acetate mixture was collected, concentrated under reduced pressure and lyophilized to obtain the above-mentioned pumpkin extract.

In the pumpkin extract of the present invention, the pumpkin sample is preferably selected from the stem of a pumpkin.

And, the use of a pumpkin extract for applying the above pumpkin extract as an insulin analog to a pharmaceutical composition for treating type 1 diabetes.

In the use of the pumpkin extract of the present invention, the above pumpkin extract may also be used as an insulin sensitizer for the preparation of a pharmaceutical composition for treating type 2 diabetes.

The above and other objects, features and advantages of the present invention will become more <RTIgt; Shown, the pumpkin extract of the present invention comprises (22E, 24R)-24-methyl-6β-methoxy-5α-cholesterol-7,22-diene-3β,5- as shown in Fig. 1. Glycol ((22E, 24R)-24-methyl-6β-methoxy-5α-cholesta-7, 22-diene-3β, 5-diol, MMCD for short) and (22E, 24R) as shown in Figure 2 3β-hydroxyergosterol-5,8,22-trien-7-one (3β-hydroxy-(22E,24R-ergosta-5,88,22-trien-7-one, abbreviated as HET), the above pumpkin extract system can be used as an insulin analogue to activate the AMPK message transmission pathway of type 1 diabetic cells. And as an insulin sensitizer, increase the sensitivity of type 2 diabetes cells to insulin, and effectively improve the absorption of glucose by cells.

Referring to Fig. 3, the pumpkin extract of the preferred embodiment of the present invention is obtained by extracting a pumpkin sample with methanol to obtain a pumpkin extract; water and ethyl acetate. The crude extract of the above pumpkin is extracted to obtain an ethyl acetate layer and an aqueous layer; and the mixture of n-hexane and ethyl acetate is used as a mobile phase, and the ethyl acetate layer fixed on a column of rubber is chromatographed. The fractions extracted with a 70-80% ethyl acetate mixture were collected, concentrated under reduced pressure, and lyophilized to obtain the above pumpkin extract.

Extracting a pumpkin sample with methanol to obtain a pumpkin crude extract, wherein the pumpkin sample is preferably selected from the stem of the pumpkin, but not limited thereto; in detail, the present invention is dry weight 71 kg of pumpkin stem powder, soaked and extracted with 120 liters of methanol, at room temperature, each soaking for 7 days, preferably three times soaking extraction, the resulting soaking solution after filtration and concentration under reduced pressure, The above crude extract of pumpkin was obtained.

The above crude extract of pumpkin is extracted and extracted with water and ethyl acetate; in a preferred embodiment of the present invention, 4 liters of water is first added to a crude extract of 7.1 kg of pumpkin, so that the crude extract of the pumpkin is suspended. Further, 4 liters of ethyl acetate was added for extraction, and it was preferred to repeat the above-described partition extraction three times to obtain the above ethyl acetate layer.

Next, the above ethyl acetate layer is subjected to chromatographic separation to obtain several regions. Separate. Specifically, the chromatographic separation is performed by using a glass column (12×150 cm), filling the normal phase tantalum as a stationary phase, and mixing one of n-hexane and ethyl acetate as a mobile phase, and the above ethyl acetate. The layer was subjected to a chromatographic separation treatment to obtain a plurality of fractions, wherein the above-mentioned n-hexane and ethyl acetate were mixed in a gradient as shown in Table 1, and n-hexane and ethyl acetate mixed in each gradient were sequentially used as a flow. The chromatographic separation process is carried out, and the segments are collected, and the fractions extracted by the above-mentioned extract containing ethyl acetate having a concentration of 70-80% (such as the fractions r, s, t or a mixture thereof) are selected. After the concentrated concentration and lyophilization, the pumpkin extract of the preferred embodiment of the present invention is obtained, wherein the pumpkin extract is preferably a fraction which is extracted with a 75% ethyl acetate extract. (such as the fraction s), obtained by concentration under reduced pressure and freeze-drying.

The fraction s can be further separated by a glass column (5×45 cm) with 350 g of colloidal rubber as a stationary phase, and extracted with a mixed solution of dichloromethane and ethyl acetate, wherein ethyl acetate accounts for 16.7~ of the mixed solution. 100%, and the distinctions sA to sH are presented in sequence.

The fraction sB was separated by high performance liquid chromatography using a liquid chromatography chromatography Si 60 column (LiChrosorb Si 60 column, 7 μm, 250 × 10 mm), with a 1:1 positive The mixture of alkane and acetone is used as a mobile phase to purify (22E,24R)-24-methyl-6β-methoxy-5α-cholesterol-7,22-diene-3β,5-diol (9 mg), the molecular formula is C ₂₉ H ₄₈ O ₃ .

Separation of sC by high performance liquid chromatography is carried out using liquid chromatography with a filler Si 60 column (LiChrosorb Si 60 column, 7 μm, 250 × 10 mm), with 4:1 dichloromethane And the ethyl acetate mixture is extracted as a mobile phase to purify (22E, 24R)-3β-hydroxyergosterol-5,8,22-trien-7-one, the molecular formula is C ₂₈ H ₄₂ O ₂ .

In order to confirm that the pumpkin extract of the preferred embodiment of the present invention can be used as an insulin analog and an insulin sensitizer, respectively, to achieve the function of lowering blood sugar in the first type of diabetic cells and the second type of diabetic cells, the following test is carried out:

(A) The above pumpkin extract reduces blood sugar levels in hyperglycemic mice

In this test, a pumpkin extract prepared by the aforementioned method was used to feed hyperglycemic mice to observe changes in blood glucose levels in the blood of hyperglycemic mice.

The hyperglycemic mice in this experiment were purchased from eight-week-old male C57BL/6JNarl mice of the National Animal Center and were housed in a temperature of 25 ± 2 ° C, humidity of 50 to 60%, light for 12 hours, and dark for 12 hours. The 17-week-old mice were firstly measured for blood glucose levels before administration (0 hours) and fasted for 2 hours and 4 hours by tail blood sampling to determine the normal blood glucose levels of the mice before induction of hyperglycemia.

The mice fasted for 8 hours were grouped into the second table, and the STZ solution (containing 10 mM sodium citrate) was intraperitoneally injected to induce the mice to become hyperglycemic mice, and the mice were rested for one week after five days of continuous administration. Then, the blood glucose values of fasting for 2 hours and 4 hours were measured once a week for three consecutive weeks by tail blood sampling to determine the high blood sugar level of the above hyperglycemic mice.

The hyperglycemic mice were given the same day before the start of the experiment, and the hyperglycemic mice were not fasted for 15 hours (ie, the feed was provided from 6 o'clock every night, and the feed was collected at 9 o'clock the next morning). Mouse weight and blood measurement The sugar value was re-fed with the pumpkin extract of the invention (200 mg/kg, dissolved in 0.5% methylcellulose) or the control group (with the same amount of methylcellulose) once a day for three consecutive days. After the drug was administered, the rats were fasted, and blood glucose was measured at the tail before the administration (0 hours) and fasting for 2 hours and 4 hours. Observing whether the pumpkin liquid has immediate or short-term improvement of hyperglycemia in hyperglycemic mice effect.

Referring to Figures 4a and 4b, the feeding of the pumpkin extract of the present invention can significantly reduce the blood glucose level of hyperglycemic mice after 2 hours, and can maintain a stable blood glucose level after 4 hours (Fig. 4b) , * or ** represents a statistically significant difference [by one-way analysis of variance (ANOVA) analysis] compared to 0 hours on the same day.

(B) MMCD and HET promote the consumption of glucose by normal cells

In order to confirm the active substances MMCD and HET of the pumpkin extract in the present case, the blood glucose concentration in the normal cells (i.e., type 1 diabetic cells) can be lowered, and the above active substances are added to the first type of diabetic cells, and the first type of diabetic cells are measured. Absorption of glucose.

In detail, this experiment used mouse liver cells (FL83B, purchased from Food Industry Development Research Institute, number BCRC 60325), and the above mouse liver cell line was cultured in 10% fetal bovine serum (Fetal Bovine Serum, abbreviated FBS) Kaighn's modification of Ham's F12K medium (referred to as F12K, purchased from Sigma-Aldrich, St. Louis, MO, USA; model N3520), and placed in a 37 ° C 5% CO ₂ incubator for subculture, After removing the old medium, the cells were washed twice with phosphate buffered saline (PBS), and serum-free F12K (100% F12K) was added for cell starvation for 3 hours. Next, the medium was removed, and washed twice with 1 time phosphate buffer, and then 450 μL of MEM medium supplemented with the additive shown in Table 3 was added, and 0, 1, 2 after the addition of the MEM medium. At 3, 4, and 5 hours, 30 μL of each medium was taken out, centrifuged at 3000 × g for 5 minutes, 10 μL of the supernatant was placed in a 96-well plate, and 250 μL of Glucose GOD FS was added, and the reaction was carried out at 37 ° C in an incubator. Minutes were measured for absorbance at 500 nm using an ELISA reader (Spectra max Plus 384, Molecular Devices LLC, USA).

Please refer to Figures 5a and 5b. Regardless of whether it is MMCD or HET, the effect is similar to that of insulin, which can significantly increase the glucose absorption of type 1 diabetes cells (such as groups B1-3 and B2-3), indicating MMCD and HET acts as an insulin analogue to boost glucose in type 1 diabetic cells Absorption amount to reduce the blood sugar of type 1 diabetes (in Figure 5a, * represents a significant difference from B1-1, statistically (by two-way ANOVA analysis); in Figure 5b, * represents and The B2-1 comparison was statistically significant (by two-way ANOVA analysis).

(C) MMCD and HET activate AMPK message delivery path

It is known that AMPK can directly catalyze the phosphorylation of AS160, and promote the transfer of GLUT4 to the cell membrane, thereby increasing the absorption of glucose by cells, and then testing whether MMCD and HET can activate AMPK and its downstream signaling pathway.

In this test, the additive shown in Table 4 was added to the culture medium, and the type 1 diabetic cells were treated for 30 minutes to prepare a crude cell protein extract, and the AS160 and ACC-1 were analyzed by Western blotting method. The total protein expression of AMPK and the phosphorylated protein expression were used to analyze IRS tyrosine phosphorylation and Akt activation in the insulin signaling pathway.

Please refer to Figures 6a and 6b. Insulin can significantly increase the phosphorylation of AMPK and the activation of AS160 and ACC-1 (Acetyl-CoA Carboxylase 1, the receptor for AMPK) in the downstream signaling pathway, while 20 μM Both MMCD and HET can promote the phosphorylation of AS160 (such as groups C1-4 and C1-6); however, if 30 μM of complex C is added to the medium (6-[4-(2-Piperidin-1-yl) -ethoxy)-phenyl]-3-pyridin-4-yl-pyrr azolo[1,5-a]-pyrimidine, referred to as compound C, is an inhibitor of AMPK), MMCD and HET are not effective in promoting phosphorylation of AS160 ( For example, CMCD and HET can also significantly activate AMPK and increase the phosphorylation of ACC-1, while the addition of complex C significantly inhibits the activation of AMPK by MMCD and HET (AMPK and ACC). -1 phosphorylation was significantly reduced), confirming that complex C can effectively inhibit the activation of AMPK, which in turn affects the activation of proteins AS160 and ACC-1 in the downstream signaling pathway.

In addition, in order to observe whether the complex C affects the effect of MMCD and HET on the collection of glucose by the cells, the additive shown in the fifth table is added to the medium, and the type 1 diabetic cells are treated for 5 hours. And 30 μL of the medium was taken out at 0, 1, 2, 3, 4, and 5 hours after the treatment, centrifuged at 3000 × g for 5 minutes, and 10 μL of the supernatant was placed in a 96-well plate, and 250 was added. μL Glucose GOD FS was reacted in a 37 ° C incubator for 10 minutes, and the absorbance at 500 nm was set by an ELISA reader to measure the amount of glucose absorbed by the cells.

Table 5: Additives applied to each group of the test

Please refer to Figures 6c and 6d. The addition of Complex C not only reduces the glucose uptake activity of insulin-enhanced cells, but also significantly reduces MMCD (such as Group C2-6) and HET (such as Group C3-6). Increased glucose uptake activity of type 1 diabetic cells (in Figure 6c, * represents a statistically significant difference (by two-way ANOVA analysis) compared to C2-1; in Figure 6d, * represents comparison with C3-1 Statistically (with two-way ANOVA analysis) there are significant differences).

Accordingly, the active substances MMCD and HET contained in the pumpkin extract of the present invention pass through the activated AMPK to activate the protein AS 160 in the downstream message transmission pathway, thereby effectively enhancing the absorption of glucose by the first type of diabetic cells. Receive, to achieve the effect of reducing blood sugar in type 1 diabetes.

(D) MMCD promotes glucose consumption by insulin-resistant cells

Treatment of FL83B cells with TNF-α for 5 hours allowed the cells to produce insulin resistance. Therefore, this experiment treated FL83B cells with TNF-α to induce insulin resistance in FL83B cells as a type 2 diabetes cell model. And in the second type of diabetic cells, insulin and a plurality of active substances in the pumpkin extract of the present invention are simultaneously added to test whether the above several active substances can promote the absorption of glucose by the second type of diabetic cells.

Specifically, this assay activates the NFκB pathway by TNF-α in FL83B cells, which produces an inflammatory response and produces insulin resistance in FL86B cells, in which cells that do not produce insulin resistance act as normal cell models and produce insulin resistance. The cells are used as a type 2 diabetes cell model; and the above-mentioned type 1 diabetes cells and type 2 diabetes cells are respectively added with additives as shown in Table 6, and the cells are treated for 5 hours and processed. After 0, 1, 2, 3, 4, and 5 hours, 30 μL of each medium was taken out, centrifuged at 3000 × g for 5 minutes, and 10 μL of the supernatant was placed in a 96-well plate, and 250 μL of Glucose GOD FS was added. The reaction was carried out in a 37 ° C incubator for 10 minutes, and the absorbance at 500 nm was set by an ELISA reader to measure the amount of glucose absorbed by the first type of diabetic cells and the second type of diabetic cells.

Referring to Figure 7, type 2 diabetic cells (such as group D1-3) do lose sensitivity to insulin (addition of insulin does not increase the amount of glucose absorbed by cells), while when the second type of diabetes cells At the same time, insulin and MMCD (such as group D1-4) or HET (such as group D1-5) were treated, it was found that MMCD can significantly promote the absorption of glucose by cells, indicating that MMCD has hypoglycemic effect on insulin-resistant cells. On the contrary, HET lacks the effect of lowering blood glucose on insulin-resistant cells (in Figure 7, * represents a significant difference from D1-1, statistically (by two-way ANOVA analysis)).

(E) Effect of MMCD on insulin signaling pathway in insulin-resistant cells

In order to test whether the MMCD can activate the insulin signaling pathway in the type 2 diabetic cells, the above-mentioned type 1 diabetes cells and the second type diabetes cells are respectively added with an additive as shown in Table 7, for the above cells. The treatment was carried out for 30 minutes, and it was observed whether several proteins (such as IRS1, Akt and AS160) of the insulin signaling pathway in the above cells were activated.

Please refer to Figure 8. Although MMCD alone cannot activate IRS1 and Akt in type 2 diabetic cells, it can significantly increase the phosphorylation of AS160 (such as group E1-5) because MMCD is permeable. Activation of AMPK enhances phosphorylation of AS160, and it is confirmed that MMCD can be used as an insulin analogue. When MMCD and insulin are simultaneously treated in type 2 diabetic cells, phosphorylation of AS160, IRS1 and Akt can be observed to increase significantly (eg, E1- Group 6), indicating that MMCD can also act as an insulin sensitizer to improve the sensitivity of insulin-resistant cells to insulin.

Accordingly, the active substance MMCD contained in the pumpkin extract of the present invention can be used as an insulin sensitizer in the second type of diabetic cells to increase the sensitivity of the insulin-resistant cells to insulin, and to increase the above-mentioned type 2 diabetes cell pair. The absorption of glucose, in turn, achieves the effect of lowering blood sugar.

The pumpkin extract of the present invention can be used as an insulin analog to activate the AMPK message transmission pathway of the first type of diabetic cells to promote the absorption of glucose by the first type of diabetic cells; the above pumpkin extract can also be used as an insulin sensitizer To improve the sensitivity of type 2 diabetes cells to insulin, and to activate insulin to activate the downstream AMPK message transmission pathway to promote the absorption of glucose by type 2 diabetes cells. Preferably, the above pumpkin extract is used alone or in combination with at least one of various ways. Pharmaceutically acceptable adjuvants, carriers, other sub-ingredients, nutrients or other active ingredients of the drug, providing raw The individual takes orally and absorbs the active ingredient of the pumpkin extract; the pumpkin extract of the present invention can be prepared into various convenient dosage forms by any food processing method, such as a tablet, a capsule, a powder, a granule or a liquid. Or combine pumpkin extract with other foods or beverages to suit the use of various biological individuals.

In summary, the pumpkin extract of the present invention has a natural component which can promote the absorption of glucose by the first type of diabetic cells and the second type of diabetic cells, thereby effectively lowering blood sugar and treating the first type and the second type of diabetes. The active ingredient; and, because the present invention and the pumpkin extract are obtained from the pumpkin through a simple step, the ingredients are natural, and as an active ingredient for treating type 1 and type 2 diabetes, it does not cause other side effects. The efficacy of the invention.

Furthermore, the pumpkin extract of the present invention can be prepared by a simple extraction step and can effectively lower the blood sugar level in the blood, and is therefore suitable for the preparation of a medicament for treating a diabetic patient, which is an effect of the present invention.

While the invention has been described in connection with the preferred embodiments described above, it is not intended to limit the scope of the invention. The technical scope of the invention is protected, and therefore the scope of the invention is defined by the scope of the appended claims.

Figure 1: Activity of the pumpkin extract of the present invention (22E, 24R)-24-methyl-6β-methoxy-5α-cholesterol-7,22-diene-3β,5-diol (MMCD) Change Learn the structure chart.

Fig. 2 is a chemical structural diagram of the activity (22E, 24R)-3β-hydroxyergosterol-5,8,22-trien-7-one (HET) of the pumpkin extract of the present invention.

Fig. 3 is a flow chart showing the extraction method of the pumpkin extract of the present invention.

Figure 4a: A line chart of changes in blood glucose concentration in hyperglycemic mice after hyperglycemia in a control group.

Figure 4b: A line graph showing changes in blood glucose concentration in hyperglycemic mice after hyperglycemia mice were fed the pumpkin extract of the present invention.

Figure 5a: Schematic diagram of reducing the glucose uptake of type 1 diabetes cells by the active substance MMCD of the pumpkin extract of the present invention.

Figure 5b: Schematic diagram of reducing the glucose uptake of type 1 diabetes cells by the active substance HET of the pumpkin extract of the present invention.

Figure 6a: Effect of the active extract of the pumpkin extract of the invention on phosphorylation of AS160 in type 1 diabetic cells.

Figure 6b: Effect of the active extract of the pumpkin extract of the invention on phosphorylation of ACC-1 and AMPK in type 1 diabetic cells.

Figure 6c: Schematic diagram of the MMCD of the pumpkin extract of the present invention increases the glucose uptake of type 1 diabetes cells.

Figure 6d: Schematic diagram of the active substance HET of the pumpkin extract of the present invention to increase the glucose uptake of the first type of diabetic cells.

Fig. 7 is a schematic view showing the activity of the pumpkin extract of the present invention for reducing the glucose uptake of the type 2 diabetic cells.

Figure 8: Signal transduction pathway of the active extract of the pumpkin extract of the present invention in type 2 diabetic cells.

Claims (3)

The use of a pumpkin active ingredient (22E, 24R)-24-methyl-6β-methoxy-5α-cholesterol-7,22-diene-3β,5-diol is used for preparation for treatment An insulin analogue of type 1 diabetes. The use of a pumpkin active ingredient (22E, 24R)-3β-hydroxyergosterol-5,8,22-trien-7-one for the preparation of an insulin analogue for the treatment of type 1 diabetes. The use of a pumpkin active ingredient (22E, 24R)-24-methyl-6β-methoxy-5α-cholesterol-7,22-diene-3β,5-diol is used for preparation for treatment An insulin sensitizer for type 2 diabetes.