Abstract:
The application of pentacyclic triterpenoid saponins, extracted from the plant of  Silene viscidula,  is used to manufacture anti-diabetic medicaments. Having screened and researched the plants and the compounds massively, the compounds of pentacyclic triterpenoid saponins extracted and purified from  Silene viscidula  (hereinafter called Wacao saponnins), have strong effects on hyperglycemia, especially for the compounds that have mother nuclei of sinocrassulosides, which can be used to manufacture anti-diabetic medicaments.

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
       [0001]    See Application Data Sheet. 
       STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT 
       [0002]    Not applicable. 
       THE NAMES OF PARTIES TO A JOINT RESEARCH AGREEMENT 
       [0003]    Not applicable. 
       INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ON A COMPACT DISC OR AS A TEXT FILE VIA THE OFFICE ELECTRONIC FILING SYSTEM (EFS-WEB) 
       [0004]    Not applicable. 
       STATEMENT REGARDING PRIOR DISCLOSURES BY THE INVENTOR OR A JOINT INVENTOR 
       [0005]    Not applicable. 
       BACKGROUND OF THE INVENTION 
       [0006]    1. Field of the Invention 
         [0007]    This invention relates to a new use of Wacao saponins and their compositions to manufacture hypoglycemic agent. The invention involves an anti-diabetic agent, an anti-diabetic composition containing the anti-diabetic agent, and a method for the treatment of diabetes in particular application. 
         [0008]    2. Description of Related Art Including Information Disclosed Under 37 CFR 1.97 and 37 CFR 1.98. 
         [0009]    Diabetes mellitus is a glucose metabolic disorder characterized by abnormally high blood glucose level (hyperglycemia) in the fasting state or during an oral glucose tolerance tests (OGTT). Generally speaking, diabetes has two major types: type 1 diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM). T1DM results from the decreased insulin and even the failure to produce insulin, a hormone which is responsible for metabolism and utilization of glucose, and whose deficiency inevitably leads to hyperglycemia. T2DM results from insulin resistance (IR), which usually, as a result, shows hyperinsulinemia, the high level of insulin in blood. IR means the main insulin sensitive tissues including liver, muscle and adipose tissues produce resistance to insulin stimulation in glucose and lipid metabolism. The consequence of IR is that the body has to secrete more insulin to compensate for IR; nevertheless, blood glucose level still increases abnormally. 
         [0010]    At present, there are several anti-diabetic drugs to choose besides insulin. Insulinotropic sulfonylurea is widely used among the chemical drugs, which has the effect on anti-diabetes via stimulating the pancreatic β-cells to secrete more insulin to increase serum insulin levels, but it has the risk of causing patients hypoglycemia. Another widely used hypoglycemic agent is biguanides, such as metformin and phenformin. Its advantages are promoting the body&#39;s peripheral glucose utilization and amending the body&#39;s high blood glucose level, but will not increase the risk of hypoglycemia. It can be used in combination with insulin or insulin secretagogues, and the side effects of which are causing lactic acidosis, diarrhea and nausea. The other relatively new class of antidiabetic drugs are insulin sensitizer glitazones (thiazolidinediones, TZDs) such as rosiglitazone and pioglitazone. They can increase the sensitivity of tissues to insulin, and reduce the levels of fasting blood glucose and insulin and the postprandial by enhancing cells&#39; glucose utilization, which, however, also have some adverse reactions including causing sodium retention, increasing blood volume, and adding cardiac load etc. 
         [0011]    Plant-derived compound screening is an important way to find new antidiabetic drugs. Among them, saponins are worthy of attention. They can prevent and treat the diabetes by regulating blood lipids, improving IR, lowering blood glucose level and so on. Current studies show that  Panax  saponins,  prunella  triterpenoid saponins,  ginseng  saponins, oleanolic saponins, bitter saponins, and aralia saponins have a good hypoglycemic effect and can be developed into new antidiabetic drugs. 
         [0012]      Silene viscidula  (Called Wacao in Chinese) is a species of plant belongs to the family Caryophyllaceae, the root of which is known as ‘Wacao’ by the hmong people in China. Wacao has the effects of analgesia, hemostasia, clearing heat and diuretic, and it is often used to treat traumatic injury, rheumatic pain, bronchitis, and urinary tract infection in clinical application. Currently, researches of  Silene viscidula  are mainly concentrated on its chemical compositions, but barely in the pharmacological. The main chemical compositions of  Silene viscidula  are saponins, proteins, organic acids, polysaccharides, and cyclic peptides etc. Our former work had successfully isolated the saponins of sinocrassuloside VI, sinocrassuloside VII, sinocrassuloside VIII, sinocrassuloside IX, sinocrassuloside XII, and sinocrassuloside XIII. The cyclic peptide composition in  Silene viscidula  mainly comprises cyclic peptides A, B, C (silenins A, B, C). The steroidal ketones components in  Silene viscidula  mainly include 20-hydroxyecdysone, 1-epi-integristeroneA, abutasterone, stachysterone A, 15-hydroxystachysterone A. The organic acid constituents in  Silene viscidula  often include hydroxycinnamic acid, oleanolic acid and vanilla acid. 
       BRIEF SUMMARY OF THE INVENTION 
       [0013]    The present invention is to solve the current technology problem by providing a kind of horizontal shaft fuel tank with charcoal canister. 
         [0014]    What the invention is intended to solve on the techniques is to provide the application of pentacyclic triterpenoid saponins extracted from  silene viscidula  and the compounds as herein defined for manufacturing medicaments possessed anti-diabetic actives. Through mass screening and research, we had firstly discovered that the compounds of pentacyclic triterpenoid saponins, especially the compound based on the nucleus structure of sinocrassuloside and its analogues, extracted and purified from a plant of  silene viscidula  have a strong hypoglycemic effect. Such compounds as herein defined and its analogues or compositions can be used to prepare the anti-diabetic drugs. 
         [0015]    The analogues of compounds comprised of the mother nucleus structure of sinocrassulosides mainly refer to the sinocrassuloside modifications and its derivatives that mainly result from aglycone of sinocrassuloside binding different numbers and combinations of glucose. These compounds include sinocrassuloside VI, sinocrassuloside VII, sinocrassuloside VIII, sinocrassuloside IX, sinocrassuloside X, sinocrassuloside XI, sinocrassuloside XII, sinocrassuloside XIII and their said pharmaceutically acceptable salts. 
         [0016]    Among them, compounds of sinocrassuloside VI and sinocrassuloside VII, sinocrassuloside VIII and sinocrassuloside IX, sinocrassuloside XII and sinocrassuloside XIII are three pairs of cis and trans isomers, all of which have good anti-diabetic activities and exhibit a certain quantitative structure-activity relationship. 
         [0017]    The said ‘pharmaceutically acceptable salt’ in the present invention means compounds of the sinocrassuloside forming salts by alkali or alkaline earth metal. The alkaline includes sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, sodium bicarbonate, ammonium chloride or ammonia; and the alkaline earth metals include sodium, potassium, calcium, aluminum, copper, zinc or magnesium. 
         [0018]    The protected pentacyclic triterpenoid compounds of Wacao saponins and their analogues in the present invention mainly refer to the compounds which is extracted and purified from a plant of  silene viscidula.  In addition, compounds of Wacao saponins and its derivatives extracted from other plants, obtained by chemical synthesis, semi-synthetic or biological transformation are also within the scope of the present invention. 
         [0019]    The said Wacao saponin compounds are mainly suitable for the treatment of T2DM, but the function is not limited to this. The dose ranges from 0.1 mg/kg to 10 mg/kg body weight. 
         [0020]    The present invention provides the use of Wacao saponin herein defined and its analogues in the manufacture of hypoglycemic medicaments. The said Wacao saponins comprise as an active ingredient compound of general formula (A): 
         [0000]    
       
                 
         
             
             
         
       
     
         [0021]    In the general formula (A): 
         [0022]    R 1 ═H, Ac, Glc or any other organic group; 
         [0023]    R 2 =(E)-MC, (Z)-MC, or Ac;
   R 3 ═H, or Xyl;   R 4 ═H, CH 3 , or CH 2 CH 2 CH 2 CH 3 ;   
 
         [0026]    The AC herein defined refers to a group as follows: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0000]    The (E)-MC herein defined refers to a group as follows: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0000]    The (Z)-MC herein defined refers to a group as follows: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0027]    According to a further embodiment, the invention concerns as follows: 
         [0028]    According to a further aspect, the said compound of Wacao saponin is represented by formula (1), hereinafter referred to as ‘Compound (1)’, the sinocrassuloside VI: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0029]    According to a further aspect, the said compound of Wacao saponins is represented by formula (2), hereinafter referred to as ‘Compound (2)’, the sinocrassuloside VII: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0030]    According to a further aspect, the said compound of Wacao saponin is represented by formula (3), hereinafter referred to as ‘Compound (3)’, the sinocrassuloside VIII: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0031]    According to a further aspect, the said compound of Wacao saponin is represented by formula (4), hereinafter referred to as ‘Compound (4)’, the sinocrassuloside IX: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0032]    According to a further aspect, the said compound of Wacao saponins is represented by formula (5), hereinafter referred to as ‘Compound (5)’, the sinocrassuloside X: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0033]    According to a further aspect, the said compound of Wacao saponin is represented by formula (6), hereinafter referred to as ‘Compound (6)’, the sinocrassuloside XI: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0034]    According to a further aspect, the-said compound of Wacao saponin is represented by formula (7), hereinafter referred to as ‘Compound (7)’, the sinocrassuloside XII: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0035]    According to a further aspect, the said compound of Wacao saponin is represented by formula (8), hereinafter referred to as ‘Compound (8)’, the sinocrassuloside XIII: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0036]    According to a further aspect, the said compounds of Wacao saponins may be used alone, a mixture of two or more, or mixed with other auxiliary materials into preparations which herein defined include injections, topical solutions, ointments, pastes, patches, drops, mouthwash, suppositories, sublingual tablets, paste film, aerosols, effervescent tablets and pills. 
         [0037]    According to a further aspect, the said injections include intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection, and intraperitoneal injection and the like. 
         [0038]    According to a further aspect, the said topical solution refers to lotion or liniment. 
         [0039]    According to a further aspect, the said ointment herein defined refers to ointment or plaster. 
         [0040]    According to a further aspect, the said drops refer to eye drops or nose drops. 
         [0041]    Still according to a further aspect, the said compounds of Wacao saponins may be used alone, a mixture of two or more, or mixed with other auxiliary materials for manufacturing medicaments, foodstuffs and drinks with the anti-diabetic activity. 
       BENEFICIAL EFFECTS OF THE INVENTION 
       [0042]    The present invention provides an application of hypoglycemic drugs made from triterpenoid saponins, especially for the compounds comprising the mother nucleus structure of sinocrassuloside and the combination of above compounds. These compounds have the significant anti-diabetic activity, and can effectively lower the blood glucose and/or improve the body&#39;s glucose tolerance. 
         [0043]    According to a still further aspect, the present invention provides compounds of Wacao saponins, their analogues and combinations, particularly the one derived from the mother nucleus structure of sinocrassuloside, have the obvious stronger hypoglycemic effects compared with hypoglycemic compounds from other reported plants. 
         [0044]    According to a still further aspect, the present invention provides compounds of Wacao saponins, their analogues and combinations, particularly the one derived from the mother nucleus structure of sinocrassuloside, have the features of novel chemical skeleton structure compare to the present hypoglycemic drugs, simple preparation process, low pollution in production, stronger hypoglycemic effects and lower side effects. 
         [0045]    Furthermore, the present invention provides compounds of Wacao saponins, their analogues and combinations, particularly the one derived from the mother nucleus structure of sinocrassuloside, are the only-chemical drug derived from natural extracts of plants that are able to compete with insulin in hypoglycemic effects. 
     
    
     
       BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS 
         [0046]      FIG. 1  is a graph illustration of the effects of Wacao saponins on glycemia in KK Ay  mice. 
           [0047]      FIG. 2  is a graph illustration of the effects of Wacao saponins on body weight in KK Ay  mice. 
           [0048]      FIG. 3  is a graph illustration of the effects of Wacao saponins on food intake in KK Ay  mice. 
           [0049]      FIG. 4  is a graph illustration of the effects of Wacao saponins on water intake in KK Ay  mice. 
           [0050]      FIG. 5  is a graph illustration of the effects of Wacao saponins on glycemia in KK Ay  mice. 
           [0051]      FIG. 6  is a graph illustration of the effects of Wacao saponins on maintaining time of anti-hyperglycemia after withdrawal in KK Ay  mice. 
           [0052]      FIG. 7  is a graph illustration of the effects of Wacao saponins on serum insulin content in KK Ay  mice. 
           [0053]      FIG. 8  is a graph illustration of the effects of Wacao saponins on ISI in KK Ay  mice. 
           [0054]      FIG. 9  is a graph illustration of the effects of Wacao saponins on hepatic glycogen in KK Ay  mice. 
           [0055]      FIG. 10  is a graph illustration of the effects of Wacao saponins on muscle glycogen in KK Ay  mice. 
           [0056]      FIG. 11  is a graph illustration of the effects of Wacao saponins on body weight in normal ICR mice. 
           [0057]      FIG. 12  is a graph illustration of the effects of Wacao saponins on blood glucose in normal ICR mice. 
           [0058]      FIG. 13  is a graph illustration of the effects of Wacao saponins on blood glucose in T2DM rats. 
           [0059]      FIG. 14  is a graph illustration of the effects of Wacao saponins on OGTT in T2DM rats 
           [0060]      FIG. 15  is a graph illustration of the effects of Wacao saponins on hepatic glycogen in T2DM rats 
           [0061]      FIG. 16  is a graph illustration of the effects of Wacao saponins on muscle glycogen in T2DM rats. 
           [0062]      FIG. 17  is a graph illustration of the effects of Wacao saponins on GSP content in T2DM rats. 
       
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
       [0063]    Note: *vs. Control group, p&lt;0.05; **vs. Control group, p&lt;0.01;  # vs. Model group, p&lt;0.05;  ## vs. Model group, p&lt;0.01. 
       EXPERIMENTAL EXAMPLES 
       [0064]    Hereinafter, the principles and features of this invention will be illustrated by reference to examples which are only for explaining the present invention, but are not intended to limit the scope of the invention. 
       Example 1 
       [0065]    Extraction, Separation, Purification and Structural Identification of Wacao Saponins 
         [0066]    The roots of  Silene viscidula  were dried and grinded into powders. 21 kg powders were extracted with 95% and 70% ethanol three times under reflux and acquired 7 kg extracts. The following separation and purification method is according to the literature (J. Zhao, Norio Nakamura, Masao Hattori. New triterpenoid saponins from the roots of sinocrassula asclepiadea [J].  Pharmaceutical Society of Japan,  2004, 52(2):230-237). Six compounds were isolated and identified as sinocrassuloside VI, sinocrassuloside VII, sinocrassuloside VIII, sinocrassuloside IX, sinocrassuloside XII, and sinocrassuloside XIII. Among them, sinocrassuloside VI and sinocrassuloside VII, sinocrassuloside VIII and sinocrassuloside IX, sinocrassuloside XII and sinocrassuloside XIII are the three pairs of cis-trans isomers. 
         [0067]    Compound (1) and Compound (2) were obtained as white amorphous powder, ESI-MS (m/z): 1473.2 [M+Na] + , 1449.7[M−H] − ;  1 H-NMR and  13 C-NMR: Table 1. The ESI-MS (m/z) of Compound (1) and (2) showed a molecular ion at m/z 1450 corresponding to the same molecular formula C 71 H 102 O 31 . It was unambiguously identified as sinocrassuloside VI and sinocrassuloside VII on the basis of its  1 H-NMR and  13 C-NMR spectral data (Table 1). 
         [0068]    Compound (3) and (4) were obtained as white amorphous powder, ESI-MS (m/z): 1487.2 [M+Na] + , 1499.7 [M+Cl] − , 1463.8 [M−H]−.  1 H-NMR and  13 C-NMR: Table 1. The ESI-MS (m/z) of Compound (3) and (4) showed a molecularion at m/z 1464 corresponding to the same molecular formula C 72 H 104 O 31 . It was unambiguously identified as sinocrassuloside VIII and sinocrassuloside IV on the basis of its  1 H-NMR and  13 C -NMR spectral data (Table 1). 
         [0069]    Compound (7) and (8) were obtained as white amorphous powder, ESI-MS(m/z): 1529.3[M+Na] + , 1541.8[M+Cl] − , 1505.6[M−H] − ;  1 H-NMR and  13 C-NMR: Table 1. The ESI-MS (m/z) of Compound (7) and (8) showed the same molecular formula C 75 H 110 O 31.  It was unambiguously identified as sinocrassuloside XII and sinocrassuloside XIII on the basis of its  1 H-NMR and  13 C-NMR spectral data (Table 1). 
         [0000]    
       
         
               
             
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                   1 H-NMR and  13 C-NMR data of Compounds (1), (2), (3), (4), (7) and (8) 
               
             
          
           
               
                   
                 Compound (1) 
                 Compound (2) 
                 Compound (3) 
                 Compound (4) 
                 Compound (7) 
                 Compound (8) 
               
               
                   
                 δHmult (J in 
                 δHmult (J in 
                 δHmult (J in 
                 δHmult (J in 
                 δHmult (J in 
                 δHmult (J in 
               
               
                   
                 Hz) a )δC b ) 
                 Hz) a )δC b ) 
                 Hz) a )δC b ) 
                 Hz) a )δC b ) 
                 Hz) a )δC b ) 
                 Hz) a )δC b ) 
               
               
                   
                   
               
             
          
           
               
                 The aglycone moiety 
               
             
          
           
               
                 1 
                 0.81, 1.37 
                 37.8 
                 0.82, 1.37 
                 37.8 
                 0.85, 1.39 
                 37.8 
                 0.85, 1.39 
                 37.8 
                 0.86, 1.40 
                 37.7 
                 0.86, 1.40 
                 37.7 
               
               
                 2 
                 1.82, 2.09 
                 24.8 
                 1.82, 2.09 
                 24.8 
                 1.81, 2.01 
                 24.7 
                 1.81, 2.01 
                 24.7 
                 1.83, 2.02 
                 24.9 
                 1.82, 2.029 
                 24.9 
               
               
                 3 
                 3.94 
                 84.2 
                 3.94 
                 84.2 
                 4.09 
                 84 
                 4.09 
                 84 
                 4.1  
                 84 
                 4.1  
                 84.1 
               
               
                   
                 (7.8) 
               
               
                 4 
                   
                 54.3 
                   
                 54.3 
                   
                 54.3 
                   
                 55.3 
                   
                 54.2 
                   
                 54.2 
               
               
                 5 
                 1.35 
                 48.5 
                 1.35 
                 48.5 
                 1.36 
                 48.5 
                 1.36 
                 48.5 
                 1.37 
                 48.4 
                 1.37 
                 48.4 
               
               
                 6 
                 0.88, 1.37 
                 20.2 
                 0.88, 1.37 
                 20.2 
                 0.89, 1.36 
                 20.2 
                 0.89, 1.36 
                 20.2 
                 0.89, 1.38 
                 20.3 
                 0.89, 1.38 
                 20.2 
               
               
                 7 
                 1.5  
                 32.2 
                 1.5  
                 32.2 
                 1.51 
                 32.1 
                 1.51 
                 32.1 
                 1.52 
                 32.1 
                 1.52 
                 32 
               
               
                 8 
                   
                 40.5 
                   
                 40.3 
                   
                 40.4 
                   
                 40.4 
                   
                 40.4 
                   
                 40.3 
               
               
                 9 
                 1.78 
                 46.3 
                 1.78 
                 46.3 
                 1.78 
                 46.8 
                 1.76 
                 46.8 
                 1.78 
                 46.7 
                 1.77 
                 46.7 
               
               
                 10 
                   
                 35.3 
                   
                 35.3 
                   
                 35.8 
                   
                 35.9 
                   
                 35.9 
                   
                 35.8 
               
               
                 11 
                 1.91 
                 23.3 
                 1.91 
                 23.3 
                 1.91 
                 23.3 
                 1.91 
                 23.3 
                 1.92 
                 23.2 
                 1.92 
                 23.2 
               
               
                 12 
                 5.57 brs 
                 121.8 
                 5.57 br s 
                 121.8 
                 5.60 br s 
                 121.8 
                 5.60 brs 
                 121.8 
                 5.61 brs 
                 122 
                 5.61 brs 
                 122 
               
               
                 13 
                   
                 143.6 
                   
                 143.5 
                   
                 143.6 
                   
                 143.6 
                   
                 143.7 
                   
                 143.6 
               
               
                 14 
                   
                 41.5 
                   
                 41.5 
                   
                 41.5 
                   
                 41.5 
                   
                 41.6 
                   
                 41.7 
               
               
                 15 
                 1.90, 2.18 
                 35.8 
                 1.90, 2.16 
                 35.9 
                 1.95, 2.18 
                 35.8 
                 1.91, 2.18 
                 35.9 
                 1.96, 2.20 
                 35.9 
                 1.96, 2.20 
                 36 
               
               
                 16 
                 5.20 brs 
                 73 
                 5.17br s 
                 73 
                 5.22 br s 
                 73.1 
                 5.20 brs 
                 73.1 
                 5.23 brs 
                 73.1 
                 5.19br s 
                 73.2 
               
               
                 17 
                   
                 49 
                   
                 49 
                   
                 48.5 
                   
                 48.5 
                   
                 48.5 
                   
                 48.5 
               
               
                 18 
                 3.39 d 
                 40.9 
                 3.38 
                 40.9 
                 3.39 d 
                 40.9 
                 3.39 d 
                 40.9 
                 3.40 d 
                 40.9 
                 3.4  
                 40.9 
               
               
                   
                 (13.8) 
                   
                   
                   
                 (14.0) 
                   
                 (14.0) 
                   
                 (14.0) 
               
               
                 19 
                  1.34, 2.74 t 
                 47.9 
                 1.34, 2.74 t 
                 47.9 
                 1.34, 2.75 t 
                 47.9 
                 1.36, 2.75 t 
                 47.9 
                 1.36, 2.76 t 
                 48 
                 1.36, 2.76 t 
                 48 
               
               
                   
                 (14.0) 
                   
                   
                   
                 (14.4) 
                   
                 (14.4) 
                   
                 (14.4) 
                   
                 (14.4) 
               
               
                 20 
                   
                 30.6 
                   
                 30.5 
                   
                 30.5 
                   
                 30.6 
                   
                 30.5 
                   
                 30.5 
               
               
                 21 
                 1.30, 2.40 
                 35.2 
                 1.32, 2.41 
                 35.2 
                 1.30, 2.41 
                 35.2 
                 1.32, 2.41 
                 35.3 
                 1.30, 2.40 
                 35.3 
                 1.32, 2.41 
                 35.3 
               
               
                 22 
                 2.22, 2.39 
                 32.2 
                 2.20, 2.38 
                 32.2 
                 2.22, 2.40 
                 32.1 
                 2.20, 2.40 
                 32.1 
                 2.23, 2.40 
                 32 
                 2.23, 2.40 
                 32 
               
               
                 23 
                 9.84 s 
                 210.5 
                 9.84s 
                 210.5 
                 9.86s 
                 210.5 
                 9.86s 
                 210.5 
                 9.88 s 
                 210.6 
                 9.88s 
                 210.6 
               
               
                 24 
                 1.39 s 
                 10.7 
                 1.39 s 
                 10.7 
                 1.39 s 
                 10.7 
                 1.40 s 
                 10.7 
                 1.41 s 
                 10.7 
                 1.41 s 
                 10.7 
               
               
                 25 
                 0.79 s 
                 15.8 
                 0.84 s 
                 15.8 
                 0.84 s 
                 15.8 
                 0.88 s 
                 15.8 
                 0.88 s 
                 15.7 
                 0.88 s 
                 15.8 
               
               
                 26 
                 1.03s 
                 17.1 
                 1.04 s 
                 17.1 
                 1.07 s 
                 17.1 
                 1.07 s 
                 17.2 
                 1.08s 
                 17 
                 1.08 s 
                 17 
               
               
                 27 
                 1.75 s 
                 26.7 
                 1.77 s 
                 26.7 
                 1.79 s 
                 26.7 
                 1.76 s 
                 26.7 
                 1.79 s 
                 26.8 
                 1.79 s 
                 26.8 
               
               
                 28 
                   
                 175.1 
                   
                 175.1 
                   
                 175.1 
                   
                 175.1 
                   
                 175 
                   
                 175.1 
               
               
                 29 
                 0.94s 
                 33.2 
                 0.94 s 
                 33.2 
                 0.96 s 
                 33.2 
                 0.98 s 
                 33.2 
                 0.98 s 
                 33.3 
                 0.99 s 
                 33.2 
               
               
                 30 
                 0.98 s 
                 24.7 
                 0.99 s 
                 24.6 
                 1.02 s 
                 24.6 
                 1.02 s 
                 24.6 
                 1.03 s 
                 24.5 
                 1.02 s 
                 24.4 
               
             
          
           
               
                 3-O-β-D-Glucuronopyranosyl 
               
             
          
           
               
                 1′ 
                 4.88 d 
                 102.6 
                 4.88 d 
                 102.6 
                 4.86 d 
                 102.6 
                 4.86 d 
                 102.6 
                 4.87 d 
                 102.6 
                 4.88 d 
                 102.6 
               
               
                 2′ 
                 4.34 t 
                 77.4 
                 4.34 
                 77.4 
                 4.36 
                 77.4 
                 4.36 
                 77.4 
                 4.36 t 
                 77.3 
                 4.36 
                 77.3 
               
               
                 3′ 
                 4.28 t 
                 84.2 
                 4.27 
                 84.2 
                 4.29 
                 84.2 
                 4.27 
                 84.2 
                 4.30 t 
                 84.1 
                 4.3  
                 84.2 
               
               
                 4′ 
                 4.44 
                 70.2 
                 4.44 
                 70.2 
                 4.24 
                 69.9 
                 4.24 
                 69.8 
                 4.25 
                 70.1 
                 4.25 
                 70 
               
               
                 5′ 
                 4.5  
                 77 
                 4.5  
                 77 
                 4.39 
                 75.3 
                 4.39 
                 75.3 
                 4.42 
                 75.2 
                 4.42 
                 75.1 
               
               
                 6′ 
                   
                 170.4 
                   
                 170.4 
                   
                 169.5 
                   
                 169.5 
                   
                 179.8 
                   
                 179.8 
               
               
                 7′ 
                   
                   
                   
                   
                 3.72 s 
                 52.4 
                 3.72 s 
                 52.4 
                 4.31 t 
                 66.5 
                 4.33 t 
                 66.5 
               
               
                 8′ 
                   
                   
                   
                   
                   
                   
                   
                   
                 1.64 m 
                 28.3 
                 1.67 m 
                 28.4 
               
               
                 9′ 
                   
                   
                   
                   
                   
                   
                   
                   
                 1.25 m 
                 22.5 
                 1.28 m 
                 22.6 
               
               
                 10′ 
                   
                   
                   
                   
                   
                   
                   
                   
                 0.85 t 
                 14.3 
                 0.86 t 
                 14.2 
               
             
          
           
               
                 2′-O-β-D-Galactopyranosyl 
               
             
          
           
               
                 1″ 
                 5.55 d 
                 102.7 
                 5.55 d 
                 102.7 
                 5.54 d 
                 102.7 
                 5.54 d 
                 102.7 
                 5.56 d 
                 102.6 
                 5.55 d 
                 102.6 
               
               
                 2″ 
                 4.46 
                 72 
                 4.46 
                 72 
                 4.47 
                 73 
                 4.46 
                 73 
                 4.47 
                 73.1 
                 4.47 
                 73.1 
               
               
                 3″ 
                 4.14 dd 
                 74 
                 4.14 dd 
                 74 
                 4.14 dd 
                 74.9 
                 4.14 dd 
                 74.9 
                 4.15 dd 
                 74.8 
                 4.14 dd 
                 74.9 
               
               
                 4″ 
                 4.55 
                 68.9 
                 4.55 
                 68.9 
                 4.57 
                 68.5 
                 4.57 
                 68.5 
                 4.56 
                 68.4 
                 4.56 
                 68.4 
               
               
                 5″ 
                 4.02 
                 75.3 
                 4.02 
                 75.3 
                 4.02 
                 77 
                 4.02 
                 77 
                 4.02 
                 77.1 
                 4.02 
                 77.1 
               
               
                 6″ 
                 4.43, 4.50 
                 60.2 
                 4.42, 4.50 
                 60.2 
                 4.44, 4.52 
                 60.3 
                 4.44, 4.51 
                 60.3 
                 4.45, 4.51 
                 60.2 
                 4.45, 4.51 
                 60.2 
               
             
          
           
               
                 3′-O-β-D-Xylopyranosyl 
               
             
          
           
               
                 1″′ 
                 5.31 d 
                 103.5 
                 5.31 d 
                 103.5 
                 5.30 d 
                 103.6 
                 5.30 d 
                 103.6 
                 5.32 d 
                 103.6 
                 5.32 d 
                 103.6 
               
               
                 2″′ 
                 3.94 t 
                 73.8 
                 3.94 t 
                 73.8 
                 3.94 t 
                 74.1 
                 3.93 t 
                 75.3 
                 3.95 t 
                 74.2 
                 3.94 5 
                 74.2 
               
               
                   
                   
                   
                   
                   
                 (7.8) 
                   
                 (8.4) 
               
               
                 3″′ 
                 4.08 
                 77.4 
                 4.08 
                 77.4 
                 4.08 
                 77.4 
                 4.07 
                 77.4 
                 4.09 
                 77.4 
                 4.09 
                 77.4 
               
               
                 4″′ 
                 4.1  
                 70 
                 4.1  
                 70 
                 4.1  
                 68.9 
                 4.09 
                 68.9 
                 4.1  
                 69.1 
                 4.1  
                 69.1 
               
               
                 5″′ 
                 3.57, 4.21 
                 66.2 
                 3.57, 4.20 
                 66.2 
                 3.64, 4.21 
                 66.2 
                 3.62, 4.21 
                 66.2 
                 3.66, 4.22 
                 66.3 
                 3.66, 4.22 
                 66.2 
               
             
          
           
               
                 28-O-β-D-Fucopyranosyl 
               
             
          
           
               
                 1″″ 
                 6.17 d 
                 93.2 
                 6.13d 
                 93.1 
                 6.19d 
                 93.2 
                 6.14 d 
                 93.2 
                 6.20 d 
                 93.3 
                 6.15d 
                 93.2 
               
               
                   
                 (8.4) 
                   
                 (8.4) 
                   
                 (7.8) 
                   
                 (7.8) 
                   
                 (8.4) 
                   
                 (8.4) 
               
               
                 2″″ 
                 4.71 t 
                 71.3 
                 4.62 t 
                 71.3 
                 4.72t 
                 70.5 
                 4.62 t 
                 70.6 
                 4.73 t 
                 70.5 
                 4.70 t 
                 70.5 
               
               
                   
                 (8.4) 
                   
                 (8.4) 
                   
                 (9.6) 
                   
                 (9.0) 
                   
                 (9.0) 
                   
                 (9.0) 
               
               
                 3″″ 
                 5.68 dd 
                 73.1 
                 5.68 dd 
                 73.1 
                 5.70 dd 
                 73.8 
                 5.69dd 
                 73.8 
                 5.72 dd 
                 73.9 
                 5.72dd 
                 74 
               
               
                   
                 (9.3, 4.8) 
                   
                 (9.3, 4.8) 
               
               
                 4″″ 
                 5.75 
                 70.1 
                 5.75 
                 70.1 
                 5.75 
                 69.9 
                 5.77 
                 69.8 
                 5.76 
                 69.8 
                 5.76 
                 69.8 
               
               
                 5″″ 
                 4.2  
                 69.9 
                 4.21 
                 68.5 
                 4.19 
                 68.5 
                 4.2 
                 68.5 
                 4.21 
                 68.5 
                 4.21 
                 68.5 
               
               
                 6″″ 
                 1.22 d 
                 16.1 
                 1.19 d 
                 16.1 
                 1.24 d 
                 16.1 
                 1.21 d 
                 16.1 
                 1.26 d 
                 16.3 
                 1.25 d 
                 16.3 
               
               
                   
                 (6.6) 
                   
                 (6.6) 
                   
                 (6.0) 
                   
                 (6.6) 
                   
                 (6.6) 
                   
                 (6.6) 
               
             
          
           
               
                 2″″-O-α-L-Rhamnopyranosyl 
               
             
          
           
               
                 1″″′ 
                 5.76 s 
                 101.6 
                 5.75s 
                 101.6 
                 5.77 s 
                 101.7 
                 5.75s 
                 101.7 
                 5.78 s 
                 101.7 
                 5.76s 
                 101.8 
               
               
                 2″″′ 
                 4.52 
                 70.5 
                 4.52 
                 70.5 
                 4.54 
                 70.2 
                 4.53 
                 70.2 
                 4.54 
                 70.4 
                 4.54 
                 70.3 
               
               
                 3″″′ 
                 4.36 
                 70.6 
                 4.36 
                 68.5 
                 4.36 
                 71.9 
                 4.35 
                 71.9 
                 4.37 
                 72 
                 4.36 
                 72 
               
               
                 4″″′ 
                 4.23 
                 71.9 
                 4.23 
                 71.9 
                 4.24 
                 72 
                 4.24 
                 72 
                 4.24 
                 72.2 
                 4.24 
                 72.2 
               
               
                 5″″′ 
                 4.4 
                 70 
                 4.4 
                 70 
                 4.44 
                 68.9 
                 4.44 
                 68.9 
                 4.46 
                 68.7 
                 4.45 
                 68.6 
               
               
                 6″″′ 
                 1.62 d 
                 18.4 
                 1.62 d 
                 18.4 
                 1.65 d 
                 18.4 
                 1.65d 
                 18.4 
                 1.66 d 
                 18.8 
                 1.65 d 
                 18.8 
               
               
                   
                 (6.0) 
                   
                 (6.0) 
                   
                 (6.6) 
                   
                 (6.6) 
                   
                 (6.6) 
                   
                 (6.6) 
               
             
          
           
               
                 The acetyl group 
               
             
          
           
               
                 1″″″ 
                   
                 169.9 
                   
                 169.8 
                   
                 169.9 
                   
                 169.9 
                   
                 170.2 
                   
                 170.1 
               
               
                 2″″″ 
                 2.00 s 
                 20.9 
                 1.97s 
                 20.9 
                 2.01 s 
                 20.9 
                 2.00 s 
                 20.7 
                 2.02 s 
                 21 
                 2.00s 
                 21 
               
             
          
           
               
                 The para-methoxycinnamoyl group (MC) 
               
             
          
           
               
                 1″″″′ 
                   
                 166.8 
                   
                 166 
                   
                 166.8 
                   
                 166.8 
                   
                 166.8 
                   
                 166.7 
               
               
                 2″″″′ 
                 6.60 d 
                 114.7 
                 5.91 d 
                 114.8 
                 6.61d 
                 114.7 
                 5.93 d 
                 114.8 
                 6.62 d 
                 114.9 
                 5.96d 
                 114.8 
               
               
                   
                 (15.6) 
                   
                 (12.9) 
                   
                 (16.2) 
                   
                 (13.2) 
                   
                 (15.6) 
                   
                 (12.0) 
               
               
                 3″″″′ 
                 7.95 d 
                 145.9 
                 6.95 d 
                 145.9 
                 7.96 d 
                 145.9 
                 6.96 d 
                 145.9 
                 7.97 d 
                 146 
                 6.98d 
                 146 
               
               
                   
                 (15.6) 
                   
                 (12.6) 
                   
                 (16.2) 
                   
                 (13.2) 
                   
                 (15.6) 
                   
                 (12.0) 
               
               
                 4″″″′ 
                   
                 126.8 
                   
                 127.1 
                   
                 126.9 
                   
                 127.1 
                   
                 127 
                   
                 127.1 
               
               
                 5″″″′ 
                 7.53 d 
                 130.9 
                 7.96d 
                 132.7 
                 7.54 d 
                 130.9 
                 7.99d 
                 132.7 
                 7.55 d 
                 131 
                 7.98d 
                 132.8 
               
               
                 &amp; 
                 (12.6) 
                   
                 (9.0) 
                   
                 (10.2) 
                   
                 (8.4) 
                   
                 (10.8) 
                   
                 (9.0) 
               
               
                 9″″″′ 
               
               
                 6″″″′ 
                 7.00 d 
                 114 
                 6.95d 
                 114 
                 7.02 d 
                 114.8 
                 6.97 d 
                 114.8 
                 7.03 d 
                 114.9 
                 6.94d 
                 114.9 
               
               
                 &amp; 
                 (8.4) 
                   
                 (9.0) 
                   
                 (9.0) 
                   
                 (9.0) 
                   
                 (9.0) 
                   
                 (9.0) 
               
               
                 8″″″′ 
               
               
                 7″″″′ 
                   
                 161.7 
                   
                 160.8 
                   
                 161.7 
                   
                 160.8 
                   
                 161.8 
                   
                 160.8 
               
               
                 p-OCH 3   
                 3.67 s 
                 55.7 
                 3.64 s 
                 55.7 
                 3.68 s 
                 55.8 
                 3.65s 
                 55.7 
                 3.67 s 
                 55.9 
                 3.68 s 
                 55.8 
               
               
                   
               
             
          
         
       
     
       Example 2 
       [0070]    Effects of Wacao Saponins on Reduction of Glycemia in Type 2 Diabetes Mellitus (T2DM) Mice 
         [0071]    Animals and Breeding 
         [0072]    80 male KK Ay  mice (12 weeks of age, License NO. SCXK (BJ) 2009-0015, Beijing Huafukang Biology Technology Co. Ltd.) and 10 male normal C57BL/6 mice (12 weeks of age, License NO. SCXK (BJ) 2009-0015, Beijing Huafukang Biology Technology Co. Ltd.) were adaptively bred for one week, and then used in the pharmacological experiment. 
         [0073]    The breeding conditions were shown in table 2. 
         [0000]                                  TABLE 2               The breeding conditions of the experimental animals                                    Temperature   22° C. ± 2° C.           Lighting   12 hs light-dark cycle                    
A 12 h light-dark cycle means the alternate time of light and dark for 12 hs, respectively.
 
         [0074]    The animals were housed individually in wood-chip-bedded plastic cages. Germ-free and high-fat diets (Beijing Huafukang Biology Technology Co. Ltd.) were used to feed the KK Ay  mice, and the standard laboratory chows were used to feed normal C57BL/6 mice. All the animals had free access to food and water. 
         [0075]    Experimental Design 
         [0076]    80 KK Ay  mice were adaptively bred for 1 week, and then randomly assigned into 8 groups: Model group, Compound (1) group, Compound (2) group, Compound (3) group, Compound (4) group, Compound (7) group, Compound (8) group and metformin group, 10 mice per group. 10 normal C57BL/6 mice were used as the control group. KK Ay  mice were fed with the high-fat diet, and the control group was fed with the normal control diet. The control and model groups were hypodermically injected with sterile water, 6 medicated groups were hypodermically injected with 6 kinds of Wacao saponins, the Compounds (1), (2), (3), (4), (7) and (8), and the positive group (metformin group) was irrigated per oral. All groups were administrated at 9 am for 2 weeks. The administrating volume is 10 ml/kg BW and the administration dosage is shown in Table 3. 
         [0077]    All mice of the fasting tail blood glucose level were measured weekly using One Touch ULTRA Glucometer (Life Scan Johnson and Johnson, USA). 
         [0000]    
       
         
               
             
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 The administration and dosage of the animals 
               
             
          
           
               
                   
                 Groups 
                 Drugs 
                 Dosage (mg/kg BW) 
               
               
                   
               
               
                   
                 Control group 
                 sterile water 
                 — 
               
               
                   
                 Model group 
                 sterile water 
                 — 
               
               
                   
                 Compound (1) group 
                 Compound (1) 
                 2.0 
               
               
                   
                 Compound (2) group 
                 Compound (2) 
                 2.0 
               
               
                   
                 Compound (3) group 
                 Compound (3) 
                 2.0 
               
               
                   
                 Compound (4) group 
                 Compound (4) 
                 2.0 
               
               
                   
                 Compound (7) group 
                 Compound (7) 
                 2.0 
               
               
                   
                 Compound (8) group 
                 Compound (8) 
                 2.0 
               
               
                   
                 Metformin group 
                 Metformin 
                 500 
               
               
                   
               
             
          
         
       
     
         [0078]    Medication 
         [0079]    Each compound of Wacao saponins was dissolved into sterile water and sterilized with filtration for Sc. The metformin was dissolved into sterile water for Ig. The control and model group mice were injected with sterile water, Sc. All the animals were treated at 9 am for 2 weeks. 
         [0080]    Experimental Procedures 
         [0081]    Overnight fasting tail blood glucose level was measured weekly using One Touch ULTRA Glucometer and measured for twice. 
         [0082]    Termination of Experiment 
         [0083]    Each group was administrated for 2 weeks (14 days) and then the experiment was terminated. 
         [0084]    Statistical Analysis 
         [0085]    Results are expressed as means±SD. Statistical analysis were analyzed by using SPSS 10.0 software. Data were evaluated with analysis of variance. 
         [0086]    Results 
         [0087]    Effects of Wacao Saponins on Glycemia in KK Ay  Mice 
         [0088]    The main purpose of this experiment is to observe and compare the effects of different compounds of Wacao saponins on glycemia in KK Ay  mice, and provide experimental basis for the following studies. 
         [0089]    Before administration (week 0), the KK Ay  mice were randomly divided into 8 groups according to their levels of glycemia. The average glycemia level of KK Ay  mice in model and Wacao saponins administration groups was markedly higher than that of the normal control C57BL/6 mice, the KK Ay  mice can be used as standard diabetic model animals. 
         [0090]    After administrated for 1 week, each compound of Wacao saponins showed glycemia down-regulating effect at different degrees: Compound (1) showed the best hypoglycemic activity (p&lt;0.01 vs model group), and then followed by Compound (2), Compound (4), Compound (3) and Compound (7), and the Compound (8) showed the weakest hypoglycemic activity which has no significant difference compared with that of the model group. 
         [0091]    After the Wacao saponins-treated groups administrated for 2 weeks, the glycemic level kept on decreasing (p&lt;0.01, vs. model group), and the tendency and degree of glycemia decreased in each compound treated group is the same as the week 1, as shown in  FIG. 1 . 
         [0092]    Discussion 
         [0093]    One of the key symptoms of diabetes is hyperglycemia. Constant hyperglycemia is harmful to patients&#39; tissues, organs and cells, which may cause chronic complications such as diabetic nephropathy, diabetic foot, diabetic retinopathy and peripheral neuropathy, etc. So it is a critical step for the treatment of diabetes to reduce the level of blood glucose. 
         [0094]    In this experiment, we compared the anti-hyperglycemia effects of different compounds with each other. It was indicated that Compounds (1) and (2) (cis-trans-isomers) had the strongest anti-hyperglycemia effect, followed by Compound (3) and (4) (cis-trans-isomers), and Compound (7), and the compound (8) showed no significant effect of anti-hyperglycemia compared with that of the other compounds. After one week&#39;s administration, Compound (8) showed only the trend of anti-hyperglycemia activity, but showed no statistical difference (p&gt;0.05) compared with that of model group. Significant differences had not yet appeared even after two weeks&#39; administration of Compound (8), which indicated that Compound (8) is far more inferior to that of the former 5 compounds in treating diabetes. 
         [0095]    Conclusion 
         [0096]    Wacao saponins had strong anti-hyperglycemia effect, which are the ideal active ingredients for treating diabetes. 
       Example 3 
       [0097]    The Therapeutic Effects of Wacao Saponins in T2DM Mice 
         [0098]    Animals and Breeding 
         [0099]    60 male KK Ay  mice (12 weeks of age, License NO. SCXK (BJ) 2009-0015, Beijing Huafukang Biology Technology Co. Ltd.) and 10 normal male C57BL/6 mice (12 weeks of age, License NO. SCXK (BJ) 2009-0015, Beijing Huafukang Biology Technology Co. Ltd.) were adaptively bred for one week, and then used in the following experiment. 
         [0100]    Animals were housed under the conditions as shown in table 4. 
         [0000]    
       
         
               
             
               
               
               
             
           
               
                 TABLE 4 
               
               
                   
               
               
                 The breeding conditions of the experimental animals 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Temperature 
                 22° C. ± 2° C. 
               
               
                   
                 lighting 
                 12 hs light-dark cycle 
               
               
                   
               
             
          
         
       
     
         [0101]    A 12 h light-dark cycle means the alternate time of light and dark for 12 hs, respectively. 
         [0102]    The animals were housed individually in wood-chip-bedded plastic cages. Germ-free and high-fat diets (Beijing Huafukang Biology Technology Co. Ltd.) were used to feed the KK Ay  mice. All the animals had free access to food and water. 
       Experimental Design 
       [0103]    60 KK Ay  mice were adaptively bred for 1 week, and then randomly assigned into 6 groups: Model group, Compound (1) group, Compound (2) group, Compound (3) group, Compound (4) group, and metformin group, 10 mice per group. 10 normal C57BL/6 mice were used for the control group. KK Ay  mice were in a high-fat diet, and the control normal mice were in a normal control diet. The control and model groups were hypodermically injected with sterile water, the 6 medicated groups mice were subcutaneously injected with 6 kinds of Wacao saponins, (Compounds (1), (2), (3) and (4)) respectively, and the positive group was administered metformin by gavage. All mice were administrated at 9 am for 2 weeks and the administration volume is 10 ml/kg BW. 
         [0104]    The body weight, food and water intake of each mouse was measured and recorded weekly throughout the course of the experiment. At the end of the experiment, collected the blood and separated serum, fasting blood glucose, insulin levels and insulin sensitivity index (ISI) were measured or calculated in each group. Liver, skeletal muscles of mice in each group were taken and weighed for detecting glycogen levels of the hepatic and muscles after mice being killed. 
         [0105]    The animals were treated as follows: 
         [0000]    
       
         
               
             
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 The administration and dosage of the animals 
               
             
          
           
               
                   
                 Groups 
                 Drugs 
                 Dosage (mg/kg BW) 
               
               
                   
               
               
                   
                 Control group 
                 sterile water 
                 — 
               
               
                   
                 Model group 
                 sterile water 
                 — 
               
               
                   
                 Compound (1)group 
                 Compound (1) 
                 2.0 
               
               
                   
                 Compound (2) group 
                 Compound (2) 
                 2.0 
               
               
                   
                 Compound (3) group 
                 Compound (3) 
                 2.0 
               
               
                   
                 Compound (4) group 
                 Compound (4) 
                 2.0 
               
               
                   
                 Metformin group 
                 Metformin 
                 500 
               
               
                   
               
             
          
         
       
     
         [0106]    Medication 
         [0107]    Compound (1), (2), (3) and (4) were respectively dissolved into sterile water, and sterilized with filtration for Sc. to the medication groups. The metformin was dissolved into sterile water for Ig. to the positive control group. Control and model groups animals were injected with sterile water (10 ml/kg BW), Sc. All the animals were treated daily at 9 am for 2 weeks (14 days). 
         [0108]    Experimental Procedures 
         [0109]    Measurement of food and water intake: 24 hours&#39; food and water intake were measured by gravimetry once a week. It should be paid attention to recycling leakage of small forage into the feed box to ensure the accuracy of food intake. 
         [0110]    Determination of body weight: The body weight of mice was measured and recorded once a week. 
         [0111]    Blood detection: Blood glucose was measured weekly. Overnight fasting glycemia level was measured with One Touch ULTRA Glucometer for the tail blood test. At the end of the experiment, blood serum was collected for measuring levels of fasting blood glucose and insulin, and the insulin sensitivity index (ISI) was calculated. At the same time, hepatic tissues and skeletal muscles of mice were taken out to determine levels of hepatic and muscle glycogen by detection kits. 
         [0112]    Termination of the experiment After administrating mice for 2 weeks (14 days), the experiment was terminated. 
         [0113]    Statistical Analysis 
         [0114]    Results are shown as means±SD. Statistical analysis data were analyzed by using SPSS 10.0 software. Data were evaluated with analysis of variance. 
         [0115]    Results 
         [0116]    Effects of Wacao Saponins on Body Weight in KK Ay  Mice 
         [0117]    Experimental data showed a more significant increase in body weight of model group than that of in the control group (p&lt;0.01), which was caused by obesity of KK Ay  mouse. In the meanwhile, the body weight growth in Wacao saponins-treated groups was obviously lower than that of model group, which indicated that Wacao saponins have a good activity of inhibiting the growth of body weight as shown in  FIG. 2 . Among them, Compound (1) showed the best effect of resistance on body weight growth in mice. 
         [0118]    Effects of Wacao Saponins on Food Intake in KK Ay  Mice 
         [0119]    Results showed that level of food intake in the model group was significantly higher than that of the control group. The result is well in line with clinical diabetes ‘polyphagia’ symptom. After treated with different medication for one week, the food intake of the Compound (1) group and the positive metformin group decreased obviously compared with the model group (p&lt;0.05). No statistical difference was found except the two groups above after one week treatment. 
         [0120]    Two weeks after treatment, except the Compound (4) group, mice in all groups showed obvious reduction in food intake (p&lt;0.05 or p&lt;0.01, vs. model group), which exhibited a certain time-dependent manner, as shown in  FIG. 3 . 
         [0121]    Effects of Wacao Saponins on Water Intake in KK Ay  Mice 
         [0122]    Experimental data showed that level of water intake of the model group was much more than that of the control group, which was consistent with the clinical symptom of ‘polydipsia’ in diabetes patients. After treated with different medication for 1 or 2 weeks, water intake in each treatment group decreased significantly (p&lt;0.05 or p&lt;0.01, vs. model group), except Compound (4) group in week 1, as shown in  FIG. 4 . 
         [0123]    Effects of Wacao Saponins on Glycemia in KK Ay  Mice 
         [0124]    This experiment is to observe the anti-hyperglycemia effects of Wacao saponins on KK Ay  mice during and after the period of drug treatment, respectively. 
         [0125]    After one week of the medical intervention, each treatment group, except the Compound (4) group, showed a significantly decrease (p&lt;0.01, vs. model group) in glycemia level. After two weeks of administration, the glycemia level in each compound treatment group decreased more obviously (p&lt;0.01, vs. model group). Furthermore, results indicated that all treatment groups except Compound (4) had stronger anti-hyperglycemia effects than that of metformin, as shown in  FIG. 5 . 
         [0126]    Meanwhile, inventors also investigated the maintaining time of anti-hyperglycemia effects after withdrawal of Wacao saponins. Results showed that after the mice stopped taking medication, glycemic levels of all treated groups rebound gradually to different degrees. After 2 weeks of drug withdrawal, glycemic levels of metformin-treated group rebounded to a high level which showed no significant difference compared with that of the model group. It was indicated that the metformin&#39;s hypoglycemic effect disappeared after 2 weeks of stopping medication. Although each Wacao saponins-treated group&#39;s blood glucose level also slightly rebounded, it was still significantly lower than that of the untreated group (p&lt;0.01). This hypoglycemic effect almost lasted for 3 weeks, which indicated that Wacao saponins have much longer duration in reducing hyperglycemia than metformin, as shown in  FIG. 6 . 
         [0127]    Effects of Wacao Saponins on Blood Insulin Content and Insulin Sensitivity Index (ISI) in KK Ay  Mice 
         [0128]    The experimental results showed that the insulin level of the model group was much higher than that of the control group (p&lt;0.01) accompanied with higher glycemia level at the same time, which indicated a certain degree of insulin resistance (IR) appeared in the model group, which was consistent with T2DM syndrome. After mice being treated with different compounds of Wacao saponins, the insulin level of all groups except Compound (4) increased significantly (p&lt;0.05 or p&lt;0.01, vs. model group), which indicated that Wacao saponins promoted β cells to secrete insulin, as shown in  FIG. 7 . 
         [0129]    The ISI of the model group decreased remarkably, but it obviously elevated after treated with Wacao saponins (p&lt;0.01, vs. model group), which indicated that Wacao saponins may improve the insulin sensitivity of animals or human, as shown in  FIG. 8 . 
         [0130]    Effect of Wacao Saponins on the Level of Hepatic and Muscle Glycogen in KK Ay  Mice 
         [0131]    The data demonstrated that the level of hepatic glycogen and muscle glycogen of KK Ay  mice significantly reduced compared with the normal mice (p&lt;0.01). Results showed that all compounds of Wacao saponins had the obvious activities of increasing hepatic glycogen level (p&lt;0.05 or p&lt;0.01, vs. model group), but exhibited an incoordinate efficacy in increasing muscle glycogen level, Compounds (1) and (2) showed stronger efficacy in raising levels muscle glycogen (p&lt;0.01, vs. model group), but Compound (3) and (4) showed a weaker efficacy in raising level of muscle glycogen which had no statistical significant difference compared with that of the model group. 
         [0132]    The experiment demonstrated that Wacao saponins can enhance the glycogen storage, especially for the hepatic glycogen. Among them, Compound (1) showed the strongest activity of the function. It was almost comparable to metformin, as shown in  FIGS. 9 and 10 . 
         [0133]    Discussion 
         [0134]    The main symptom of diabetes is hyperglycemia, accompanied with such clinical manifestations as polydipsia, polyphagia, diuresis and athrepsy. Constant hyperglycemia will harm the body tissues, organs and cells, and lead to diabetic nephropathy, diabetic foot and eye ground, peripheral neuropathy and other chronic complications. Therefore, it is the primary goal for treating diabetes to reduce the body&#39;s glucose level. In addition to high blood sugar, Type 2 diabetes is often characterized by abnormal glucose tolerance, the drop of insulin sensitivity, lipid metabolism disorder and abnormal insulin tolerance. Laboratory tests show high levels of TC, TG and LDL, but low levels of HDL in T2DM patients. 
         [0135]    The results of this experiment indicated that Wacao saponins could effectively and efficiently improve the symptoms of polydipsia, polyphagia of diabetic model mice, decrease the blood glucose, promote the secretion of insulin, increase ISI and the contents of hepatic and muscle glycogen. Among them, Compound (1) showed the best anti-diabetic effects characterized by good hypoglycemic effect and the increase in insulin sensitivity, which is similar to the positive drug of metformin. Furthermore, its hypoglycemic effect lasts longer than that of metformin, which can effectively prevent blood sugar from rebounding in a short period of time after stopping medication. These observations collectively demonstrate that the anti-diabetic activity of Wacao saponins may be related to such functions as promoting insulin secretion, enhancing insulin sensitivity and promoting reserves of peripheral blood sugar in body&#39;s tissues. 
         [0136]    Conclusion 
         [0137]    Wacao saponins can improve the symptoms of polydipsia and polyphagia of diabetic model mice, effectively promote insulin secretion, increase ISI and enhance glycogen reserves, which indicated that Wacao saponins have a strong hypoglycemic effect with good druggability, and can be used to manufacture the ideal drugs in treating diabetes. 
       Example 4 
       [0138]    Effects of Wacao Saponins on Reducing Glycemia in Normal ICR Mice 
         [0139]    This experiment is to investigate the effect of Compounds (1) and (2) on reducing glycemia in normal ICR mice. 
         [0140]    Animals and Breeding 
         [0141]    40 male ICR mice (20-22 g, License NO. SCXK (BJ) 2011-0004, Beijing SPF Experimental Animal Technology Co. Ltd.) were adaptively bred for one week, and then used in the following experiment. 
         [0142]    Animals were housed under the following condition, as shown in Table 6 
         [0000]    
       
         
               
             
               
               
               
             
           
               
                 TABLE 6 
               
               
                   
               
               
                 The breeding conditions 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Temperature 
                 22° C. ± 2° C. 
               
               
                   
                 lighting 
                 12 hs light-dark cycle 
               
               
                   
               
             
          
         
       
     
         [0143]    A 12 h light-dark cycle means the alternate time of light and dark for 12 hs, respectively. 
         [0144]    Animals were housed individually in wood-chip-bedded plastic cages and fed the standard laboratory chow. All the animals had ad libitum access to food and water. 
         [0145]    Experimental Design 
         [0146]    40 ICR mice were adaptively bred for 1 week, and then randomly assigned into 4 groups: Model, Compound (1), Compound (2) and metformin group, 10 mice per group. All the animals were fed with normal food. Mice in the control group were hypodermically injected with sterile water. The treated groups were subcutaneously injected with Compounds (1) and (2), respectively. And the positive group was administrated metformin by gavage. All the animals were administrated at 9 am for 2 weeks, and the dose volume is 10 ml/kg BW. 
         [0147]    The body weight and fasting blood glucose were measured and recorded weekly. 
         [0148]    The animals were treated as the following: 
         [0000]    
       
         
               
             
               
               
               
               
             
           
               
                 TABLE 7 
               
             
             
               
                   
               
               
                 The administration and dosage of the animals 
               
             
          
           
               
                   
                 Groups 
                 Drugs 
                 Dosage (mg/kg BW) 
               
               
                   
               
               
                   
                 Control group 
                 sterile water 
                 — 
               
               
                   
                 Compound (1) group 
                 Compound (1) 
                 2.0 
               
               
                   
                 Compound (2) group 
                 Compound (2) 
                 2.0 
               
               
                   
                 Metformin group 
                 Metformin 
                 500 
               
               
                   
               
             
          
         
       
     
         [0149]    Medication 
         [0150]    Compound (1) and (2) were respectively dissolved into sterile water, and sterilized with filtration for Sc. The metformin was dissolved into sterile water for Ig. to the positive control group. Mice in the control group were injected with sterile water (10 ml/kg BW), Sc. All the animals were treated at 9 am for 2 weeks (14 days). 
         [0151]    Experimental Procedures 
         [0152]    Determination of body weight: Body weights of animals were weighed and recorded once a week. 
         [0153]    Blood detection: Blood glucose was measured weekly. Overnight fasting glycemia was measured by One Touch ULTRA Glucometer (LifeScan Johnson and Johnson, USA) for the tail blood test. 
         [0154]    End of the experiment: After mice being administrated for 2 weeks (14 days), the experiment was terminated. 
         [0155]    Statistical Analysis 
         [0156]    Data were expressed as mean±SD. Statistical analysis data were analyzed by using SPSS 10.0 software. Data were evaluated with analysis of variance. 
         [0157]    Results 
         [0158]    Influence of Wacao saponins on body weight in normal ICR mice 
         [0159]    There was no significant difference found in body weight between the Wacao saponins-treated and the normal control groups, which indicated that Wacao saponins have no obvious impact on body weight of ICR mice, as shown in  FIG. 11 . 
         [0160]    Effects of Wacao Saponins on Glycemia in Normal ICR Mice 
         [0161]    After mice being administrated for 1 week, the glycemia level in each Wacao saponins treated groups was decreased more or less, but no significant difference was found except the Compound (1) group (p&lt;0.05, vs. control group) compared with the control group. After mice being administrated for 2 weeks, the glycemia level of the Compounds (1) and (2) group decreased much obviously (p&lt;0.01, vs. control group), which indicated a stronger effect on anti-hyperglycemia, as shown in  FIG. 12 . 
         [0162]    Conclusion 
         [0163]    Compound (1) and (2) can decrease the normal blood glucose of ICR mice, which indicated that Wacao saponins had obvious hypoglycemic effects on normal ICR mice as well. 
       Example 5 
       [0164]    The Therapeutic Effects of Wacao Saponins in Experimental T2DM Model Rats 
         [0165]    Animals and Breeding 
         [0166]    50 male SD rats (180-200 g, License NO. SCXK (BJ) 2011-0004, Beijing SPF Experimental Animal Technology Co. Ltd.) were adaptively bred for 1 week, and then used in the pharmacological experiment. 
         [0167]    Animals were housed under the conditions as shown in table 8. 
         [0000]    
       
         
               
             
               
               
               
             
           
               
                 TABLE 8 
               
               
                   
               
               
                 The breeding conditions 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Temperature 
                 22° C. ± 2° C. 
               
               
                   
                 Lighting 
                 12 hs light-dark cycle 
               
               
                   
               
             
          
         
       
     
         [0168]    A 12 h light-dark cycle means the alternate time of light and dark for 12 hs, respectively. 
         [0169]    Animals were housed individually in wood-chip-bedded plastic cages and fed a standard laboratory chow. All the animals had ad libitum access to food and water. 
         [0170]    Experimental Design 
         [0171]    50 SD rats were adaptively bred for 1 week, and then randomly assigned into 5 groups: The control group, model group, Compound (1) group, Compound (2) group and glucobay group (the positive group), 10 rats per group. All the animals were in a standard laboratory diet. Control and model group animals were hypodermically injected with sterile water, test compound treatment groups were subcutaneously injected with the Compounds (1) and (2), and the positive group was administrated glucobay solution by gavage. All the animals were administrated at 9 am for 2 weeks, and the dose volume is 10 ml/kg BW. 
         [0172]    The body weight and fasting blood glucose level of each rat was measured weekly. 
         [0173]    The animals were treated as follows: 
         [0000]    
       
         
               
             
               
               
               
               
             
           
               
                 TABLE 9 
               
             
             
               
                   
               
               
                 The administration and dosage 
               
             
          
           
               
                   
                 Groups 
                 Drugs 
                 Dosage (mg/kgBW) 
               
               
                   
               
               
                   
                 Control group 
                 sterile water 
                 — 
               
               
                   
                 Model group 
                 sterile water 
                 — 
               
               
                   
                 Compound (1) group 
                 Compound (1) 
                 4.0 
               
               
                   
                 Compound (2) group 
                 Compound (2) 
                 4.0 
               
               
                   
                 Glucobay group 
                 Glucobay 
                 20 
               
               
                   
               
             
          
         
       
     
         [0174]    Medication 
         [0175]    The Compounds (1) and (2) were dissolved into sterile water, and sterilized with filtration, Sc. Glucobay was dissolved into sterile water, Ig. Animals in the control and model groups were injected with sterile water (10 ml/kg BW), Sc. All the animals were treated at 9 am for 2 weeks (14 days). 
         [0176]    Experimental Procedures 
         [0177]    Preparation of T2DM Animal Model 
         [0178]    All rats except the control group were administrated intragastrically with the high-fat emulsion (10 ml/kg) and weighed the body weight weekly. After weeks of administration, rats were intraperitoneally injected with streptozotocin (STZ, dissolved in citric acid-citrate sodium solution, pH 4.2, freshly prepared and used instantly, and store away from light) (30 mg/kg). 4 days later, blood glucose level in rats was measured by tail blood test. Diabetes was defined as fasting blood glucose level&gt;11.1 mmol/Lin in this experiment. 
         [0179]    Blood Glucose Measurement 
         [0180]    The blood glucose level was measured before and after administration of the treatment compounds for 2 weeks, respectively. The animals were fasted overnight and tail blood samples were taken. Fasting glycemia was measured by using One Touch ULTRA Glucometer or kits. 
         [0181]    Oral Glucose Tolerance Test (OGTT) 
         [0182]    OGTT was performed after administration for 2 weeks. Animals were fasted for 16 hours and the fasting blood glucose level was measured prior to the start of the OGTT. Then animals were treated with 50% glucose water solution at rate of 5 g/kg BW by gavage. Blood samples were collected and the blood glucose level was measured at 30, 60 and 120 minutes after the glucose load. 
         [0183]    Measurement of Glycosylated Serum Protein (GSP) and Glycogen Level of Liver and Muscles 
         [0184]    At the end of the experiment, the abdominal aorta blood was collected to determine GSP, and the liver and skeletal muscles were taken to detect glycogen. 
         [0185]    Termination of the Experiment 
         [0186]    The experiment was terminated after 2 weeks of compounds administration. 
         [0187]    Statistical Analysis 
         [0188]    Data are expressed as mean±SD. Statistical analysis data were analyzed by using SPSS 10.0 software. Data were evaluated with analysis of variance. 
         [0189]    Results 
         [0190]    Effect of Wacao Saponins on Glycemia in T2DM Rats 
         [0191]    After mice being administrated for 2 weeks, the glycemia level of Compounds (1) and (2) groups was significantly decreased compared with the model group (p&lt;0.01, vs. model group).The result indicated that Compounds (1) and (2) had a strong effect on anti-hyperglycemia. And such efficacy of Compound (1) was much stronger than that of the glucobay, as shown in  FIG. 13 . 
         [0192]    Effect of Wacao Saponins on OGTT in T2DM Rats 
         [0193]    30 minutes after the glucose load, all groups&#39; glycemia level increased, but the normal control group increased inconspicuously. The model and Wacao saponins-treated groups increased dramatically. No statistical significant difference was observed between the model and treatment groups. Glycemia level of the control group started increasing at 30 mins after glucose load, then continuously decreased to normal level at 120 mins. The glycemia level of the model group decreased slowly and maintained a higher level within 120 minutes after glucose load compared with the normal control group, which demonstrated that the glucose tolerance of the model group was impaired. By contrast, compounds-treated groups&#39; glycemia level declined significantly compared with the model group (p&lt;0.05 or p&lt;0.01). Compounds (1) and (2) showed a similar effect on improving glucose tolerance to glucobay, as shown in  FIG. 14 . 
         [0194]    Effect of Wacao Saponins on the Contents of GSP and Glycogen of Hepatic and Muscle in T2DM Rats 
         [0195]    Data demonstrated that hepatic and muscle glycogen levels of T2DM rats decreased significantly (p&lt;0.05), but the GSP level increased obviously (p&lt;0.01) compared with the normal rats. After treatment, the hepatic and muscle glycogen levels of each treatment group significantly increased, but the GSP level obviously decreased at the same time. Compounds (1) and (2) showed a better effect on up-regulating of glycogen levels and down-regulating of GSP levels in the oral glucose tolerance test than glucobay, as shown in  FIGS. 15, 16 and 17 . 
         [0196]    Discussion 
         [0197]    At present, it is a common method to prepare diabetes animal model by injecting overdose of streptozotocin (STZ) or alloxan, whose pathological mechanism is to selectively destroy pancreatic β cells and reduce the content of blood insulin, which leads to increase blood glucose level. The results reasonably account for the pathologic features of type 1 diabetes mellitus (T1DM). The features of the deficiency of insulin secretion caused by STZ or alloxan are quite different from the pathologic process and clinical features of the type 2 diabetes mellitus (T2DM). In this experiment, we used high-fat diet administration combining low pathologic dose injection of STZ to establish the T2DM animal model. 
         [0198]    Data showed that glycemia level of model group was much higher than that of the normal control group, which demonstrated that T2DM model was successfully established. After treated with Compounds (1) and (2), the glycogen level decreased and the GSP level increased in the model animals. The results demonstrated that Compounds (1) and (2) had a good anti-hyperglycemia effect on T2DM and the efficacy was stronger than glucobay. 
         [0199]    The experimental results demonstrated that hepatic and muscle&#39;s glycogen contents declined obviously in T2DM model rats compared with that of normal rats (p&lt;0.05). Fortunately, Compounds (1) and (2) could obviously elevate hepatic and muscles glycogen level in T2DM model rats. And the increase in glycogen level was even higher than that of the normal rats, which indicated that Compounds (1) and (2) had a good effect on inhibiting glycogenolysis, increasing the glycogen level, and counteracting the hyperglycemia which can cause damage to liver and peripheral tissues. Such effect of the compounds is superior to that of glucobay. 
         [0200]    GSP is a kind of high molecule ketoamine with stable chemical structure, which is formed when no enzymatic glycosylation action happens between glucose and serum protein molecule at high glucose status. As the halflife time of serum albumin is about 17 to 20 days, GSP level can reflect the average glycemia level in 2 or 3 weeks, which will not be invulnerable to glycemia content. The results showed that GSP level rose markedly in T2DM model group, but Compounds (1) and (2) down-regulated the GSP level significantly, whose effects are stronger than glucobay. 
         [0201]    Conclusion 
         [0202]    Compounds (1) and (2) have good efficacies of reducing blood glucose, improving glucose tolerance, increasing liver and muscle&#39;s glycogen level, and decreasing serum GSP content. All in all, these results indicated that Compounds (1) and (2) were certain strong hypoglycemic ingredients which could be used to manufacture anti-diabetic medicaments. 
         [0203]    The experimental examples above were only samples to illustrate the present invention, but not the limitation of the invention. Any revision, alternative substitution, or modification based on the present invention belongs to the scope of protection.