Background technology
Cardiovascular and cerebrovascular disease and diabetes are deadly diseases of serious threat human health, and the sickness rate of apoplexy, coronary heart disease, diabetes is more and more high at present, and becomes younger day by day, is the highest disease of mortality rate except that malignant tumor.At these three kinds of chronic diseases, the mankind still do not have the radical cure way at present.
Cardiovascular and cerebrovascular disease and diabetes are diseases of one group of height correlation, and same patient often suffers from the multiple disease in above-mentioned three kinds of diseases simultaneously, such as suffering from coronary heart disease and diabetes simultaneously.At present, modern medicine has realized that the generation of above-mentioned disease, development have numerous identical pathomechanisms, these pathological changes or reciprocal causation or addition each other, thus cause disease to take place.
At present, medical circle generally acknowledges that insulin resistant (IR) is cardiovascular and cerebrovascular disease and the total pathogenesis of diabetes.Body can cause a series of pathological change after producing insulin resistant because of a variety of causes, such as: obesity, hyperlipidemia, carbohydrate tolerance reduction etc.As time goes on, hypertension, atherosclerosis be will progressively produce, thereby coronary heart disease and apoplexy caused.Certainly, insulin resistant itself is exactly the early stage performance of diabetes, and its natural history must cause the generation of diabetes.
Therefore, for there being insulin resistant but as yet the crowd of morbidity not treats intervention at insulin resistant, is that the above-mentioned disease of prevention takes place essential and effective means.
If coronary heart disease, apoplexy take place, no matter be that acute stage or convalescent care have common intervention target spot too, such as: antioxidation and antiinflammatory.For acute stages treated, antioxidation and anti-inflammatory drug can reduce the death of cell, reservation function to greatest extent.For convalescent care, antioxidation and antiinflammatory and blood fat reducing can delay atherosclerotic generation, reduce the probability that thrombosis (or hemorrhage) takes place once more.
As from the foregoing, glucagon opposing, antioxidation, anti-inflammatory response and blood fat reducing are common, the essential control target spots of cardiovascular and cerebrovascular vessel and diabetes.
Regrettably, prior art still can't provide the single medicine that possesses above-mentioned four kinds of pharmacological actions simultaneously.Existing medicine often all is to intervene (treatment) at single pathomechanism (target spot).Only at hypertension, blood lipid-lowering medicine is only at hyperlipidemia such as, antihypertensive drug, and diabetes medicament is only at diabetes, or the like.Therefore, a patient often will use multiple medicine in treatment, and this not only causes patient's inconvenience of taking medicine, and even more serious is: numerous medicines tend to bring more side effect, brings difficulty also for simultaneously doctor's clinical treatment decision-making.
So there is intensive demand this area to many target drugs of cardiovascular and cerebrovascular disease and diabetes control.
The world of medicine's practice is for many years recognized people, and the unification compound is difficult to satisfy many target spots pharmacological action demand.Therefore, this area scientific research personnel begins to consider to fall multiple existing chemical compound and is combined into the compound medicine use, so that satisfy the demands.Simultaneously, also there are more and more many people to turn one's attention to " natural " medicinal plants, because natural drug has the irreplaceable advantage of chemical synthetic drug.And along with the high speed development of modern science, various compositions can be by sharp separation and evaluation in the plant amedica, and this lays a good foundation for the development of natural drug.
In recent years, many work is being done by the world of medicine aspect the medicinal plants study of this area, strengthened analysis and research to bioactive substance, in the hope of further clear and definite its effect.Yet the effect that obtains is limited.This mainly is the pathogenesis complexity owing to cardiovascular and cerebrovascular vessel and diabetes, and normal with one or more different classes of complication, existing single active skull cap components is difficult to comprehensive proving effective.
It is reported that genipin and peoniflorin be the energy blood sugar lowering all, but mechanism of action is not fully aware of separately (referring to: CELL METABOLISM, JUNE 2006; New Chinese medicine and clinical pharmacology 2006,17 (1): 1-3; Planta Med1997,63 (4): 323).Peoniflorin has effects such as antiinflammatory, blood fat reducing (referring to Chinese Pharmacological circular ChinesePharmacological B ulletin 2003 Sep; 19 (9): 965~70), about other pharmacological actions of genipin research and report are arranged seldom then, this further improves treatment of diabetes for the pluses and minuses of scientifically utilizing the two is disadvantageous.
The coupling of different pharmaceutical, as if always a kind of good solution.Yet, up to now, at genipin can with the Radix Paeoniae use in conjunction, the mechanism of action of the two is underlying issues such as adduction, collaborative or antagonism, prior art does not provide scientific basis even similar hint or attempt not.
Summary of the invention
One object of the present invention is to provide genipin and Radix Paeoniae to be used for the treatment of and/or to prevent application in the medicine of cardiovascular and cerebrovascular disease and diabetes in preparation.
Cardiovascular and cerebrovascular disease
The inventor studies show that, genipin and Radix Paeoniae unite use, obtaining synergism aspect antioxidation, antiinflammatory, the blood lipid regulation, this is the result that this area obtains with basic test for the first time.Infer its mechanism, we estimate is the free radical capture effect that genipin has specific target spot, under this effect, has strengthened the antioxidation of peoniflorin, has reduced inflammatory reaction simultaneously, and this is a common phenomena in medical pathologies physiology.
Cardiovascular and cerebrovascular disease of the present invention is meant: coronary heart disease, apoplexy, myocardial infarction, vascular senile dementia.
Diabetes
Genipin needs sizable dosage that hypoglycemic activity is just arranged, not medical value.Under the guide of a large amount of pre-stage tests, through a large amount of experimentatioies, the inventor confirms that first genipin and Radix Paeoniae can give diabetics simultaneously.After adding peoniflorin, significantly reduced the consumption of genipin when reaching identical blood sugar lowering level, made genipin aspect treatment or prevent diabetes effectively, possess medical value.
Advantageously, further studies show that: the coupling of the two is obtaining synergism aspect the solution insulin resistant, and this is that those skilled in the art are unpredictable.
Expectation is not limit by any theory or prescription, and the inventor infers that this is because Radix Paeoniae acts on different target spots with genipin, especially at different pathological stages performance therapeutical effect, has brought into play the wholistic therapy advantage of drug combination.
When clinical practice, the doctor can formulate individual dosage regimen according to factors such as patient's age, system, the orders of severity, for example gives 1/5-1 and doubly treat/prevent the pharmaceutical composition of the present invention of effective dose.
Wherein, diabetes of the present invention are meant non-insulin-dependent diabetes mellitus and insulin dependent diabetes mellitus (IDDM).
Second purpose of the present invention is that the present invention is combined in the application in the preparation functional health product.
Because the present composition possesses above-mentioned effect, also meets the requirement of functional health product safety.Therefore, combinations thereof of the present invention also can be used as the major function composition of preparation functional health-care food or is spiked in food, the beverage as accelerant, for adaptation population's life-time service.And, on the basis of the present invention's test, it will be appreciated by persons skilled in the art that the functional health product that contains combinations thereof of the present invention has following health care at least: auxiliary lipid-lowering function, auxiliary hyperglycemic function, anti-oxidation function.
Equally, the health product technology of preparing also belongs to known technology.But for medicine, the present composition is used for source, the condition of health product can be more extensive and loose.For example, can use the combination of genipin and Radix Paeoniae Alba extract, also can directly select the combination of genipin and peoniflorin, also can select genipin and contain other plant extract of peoniflorin or use complete synthesis, the semisynthetic of its chemistry.In the functional component of health product constitutes, can only contain the present invention's combination, also can be that the present invention makes up and adds that other is auxiliary or strengthen the composition of effect, as: add vitamin E in order to strengthen anti-oxidation efficacy, do not repeat them here.
Peoniflorin of the present invention, genipin can be independent dosage form for administering drug combinations, the product form that also can contain the two simultaneously provides.
Therefore, another object of the present invention is to be provided for containing the Pharmaceutical composition of peoniflorin and genipin, in wherein genipin and peoniflorin, the part by weight of the two is 0.1-10: 1, preferred 0.2-5: 1,1: 8-8: 1 and more preferably 1: 3,1: 1 and 1: 2.
In the present composition, can select flavour of a drug (Radix Paeoniae) directly to be ground into powder and be used as medicine, extract or other forms that also can be equivalent to above-mentioned Chinese crude drug crude drug amount are used as medicine.Therefore, the activity of pharmaceutical composition of the present invention is formed and is comprised the former powder of medical material, fat or water solubility extract, effective site or effective ingredient.For example, comprise described active the composition:
A) genipin (genipin): genipin, structural formula is as follows:
Genipin uses in preparation as medical artificial skin at present extensively as the albumen coupling agent.Monomer formulation also can directly be buied, so no longer narrate its preparation method at this for known technology in its preparation method and source in market; With
B) be the material of main active with peoniflorin (Paeoniflorin): the dry root powder of Radix Paeoniae (Radix Paeoniae Alba, Radix Paeoniae Rubra, river Radix Paeoniae Rubra), the extract that contains glycosides compound (being preferably peoniflorin and lactone glucoside of Radix Paeoniae) or peoniflorin monomer or derivatives thereof.In addition, studies show that the Radix Paeoniae Alba extract that further contains peoniflorin, lactone glucoside of Radix Paeoniae, Hydroxy peoniflorin, oxypaeoniflorin, benzoylpaeoniflorin and/or lacdtlorin etc. also is useful.The source that it will be appreciated by persons skilled in the art that Radix Paeoniae glycoside of the present invention is not limited to Radix Paeoniae, contains belonging to other plant together and also can realizing the present invention of Radix Paeoniae glycoside, and it extracts and preparation method is a known technology, does not give unnecessary details at this.
Advantageously, above-mentioned active component is when being used as medicine with form of extract, and the purity of peoniflorin in described extract is 5-98wt%, preferred 30%, 60%, 75% or 97%.
Can adopt modern extract and isolation technics with the purity of raising active substance, remove unwanted impurity as far as possible, similar techniques is quite effectively ripe in the prior art, for example: Chinese patent application ZL021109737, ZL021332983, ZL031131263; With ZL99114969.6, ZL99112969.5, ZL03109126.1, ZL200510026142.6.
Can be at absorption characteristics in the dissolution characteristics of Radix Paeoniae glycoside, genipin and the body, adopt the standard preparation technology to make suitable oral or parenterai administration dosage form, similar techniques is also quite ripe in the prior art effectively, for example: tablet, dispersible tablet, oral cavity quickly disintegrating tablet, sustained-release preparation, drop pill, soft capsule, stable injection or infusion solution or contain dosage form such as phospholipid preparation.
That carries out subsequently studies show that, compare with existing cardiovascular and cerebrovascular vessel and diabetes medicament combination, pharmaceutical composition of the present invention has still kept identical even more favourable pharmacological action (for example having avoided unwanted untoward reaction) with regard to some aspect, many index significantly is better than the genipin or the Radix Paeoniae of using separately, and this is that those skilled in the art can't infer according to prior art.Therefore, the present invention attempts to provide a kind of selection, and it provides a kind of new effective product as the substituting of prior art, and this product and corresponding treatment scheme thereof will be that the doctor highly approves and puts forth effort and recommends.
The pharmacological effect experimental study
1. to antioxidation and the blood lipid regulation effect of IR model hyperlipidemia rat
Material
Present composition A: genipin+peoniflorin 1: 3 (80,20mg/kg);
Present composition B: genipin+peoniflorin 3: 1 (80,20mg/kg);
Present composition C: genipin+Radix Paeoniae Alba extract (containing Radix Paeoniae Alba total glycosides 65%) 2: 1 (80,20mg/kg);
Normal control group: do not have; Hyperlipidemia group: do not have;
Contrast medicine: genipin (20mg/kg, 40mg/kg); Peoniflorin (40mg/kg, 20mg/kg);
Positive drug: rosiglitazone group: 3mg/kg.
Experimental technique
The foundation of model: after SD rat adaptability raised for 1 week, random packet, 8 every group.Normal group is fed normal feedstuff, drinks the tap water of cleaning.All the other rats feed high fat, high sugar feed (normal feedstuff 42%, Adeps Sus domestica 23%, sucrose 24%, Sal 5%), and drink 1% saline solution.Once a day, medicine is irritated stomach, gets the hematometry observation index after continuous 4 weeks.
The results are shown in following table 1,2:
The influence (nmol/L) of table 1 pair hyperlipemia rat serum and liver LPO and SOD
Compare with the hyperlipidemia group
*P<0.05
*P<0.01
The influence (mmol/L) of table 2 pair IR rat model blood fat
Compare with the hyperlipidemia group
*P<0.05
*P<0.01; Compare with normal group
△ △P<0.01
Conclusion
Hyperlipidemia model and normal group compare, and the LPO of serum and liver obviously raises, and obviously reduce and SOD is active, show that hyperlipemia rat is with the free radical lipid peroxidation injury.Simultaneously, the TC of model group, TG and HDL-C obviously raise than normal group, show that model group has the disorders of lipid metabolism symptom.
Above-mentioned experiment shows, there is certain antioxidant activity in the peoniflorin of using separately when heavy dose of, the low dosage peoniflorin is then not obvious, genipin does not then have antioxidant activity, low, the high dose group of each group of the present composition all have antioxidant activity, especially the activity of high dose group more remarkable (p<0.01) obviously is better than peoniflorin, genipin and the rosiglitazone contrast medicine used separately, shows that the two unites to use and have well collaborative anti-oxidation efficacy.Each medicine group is basic consistent with antioxidant activity for the influence performance of blood fat, demonstrates the advantage of the present composition equally.
2, to the influence of tumor necrosis factor injury rats heart microvascular endothelial cell inflammatory factor
Material
Present composition A: genipin+peoniflorin 1: 5 (25,50,100 μ g/ml);
Present composition B: genipin+Radix Paeoniae Alba extract (containing peoniflorin 50%) 5: 1 (25,50,100 μ g/ml);
Contrast medicine: genipin (50 μ g/ml, 100 μ g/ml); Peoniflorin (50 μ g/ml, 100 μ g/ml);
Method
By CMEC In vitro culture technology, set up the cell model of TNF-α damage, be evaluation index with cell viability (MTT), lactic acid dehydrogenase (LDH), observe the protective effect of medicine to the CMEC of damage.Detect the content of interleukin 6 (IL-6), monocyte chemoattractant protein 1 (MCP-1), interleukin 8 (IL-8) in the damage CMEC supernatant with the double-antibody sandwich elisa method; Detect the evaluation index that is expressed as of damaging CMEC IL-6, MCP-1, IL-8mRNA with real time quantitative PCR method, observe medicine and the CMEC that damages is synthesized the influence that discharges inflammatory mediator at variable concentrations.
The result
Show that the present composition 25~100 μ g/ml all can improve the cell viability of TNF-α damage CMEC, reduce the release of LDH, with 50 μ g/ml best results.Genipin does not have this effect, and peoniflorin 100 μ g/ml have faint effect, and 25 μ g/ml are suitable with the present composition.
The present composition can concentration dependent in 25~100 μ g/ml scopes the release of inhibition TNF-α damage CMECIL-6 and the expression of mRNA.Genipin does not have this effect, and peoniflorin 100 μ g/ml have faint effect, but is weaker than the present composition 25 μ g/ml slightly.
The present composition is to all reducing the release of IL-6 in TNF-α damage CMEC effect 3~12h, wherein effect is the most obvious when 9h.The MCP-1 of inhibition TNF-α damage CMEC that can concentration dependent in 25~100 μ g/ml scopes discharges, and effect growth inhibitory action in time weakens gradually.The present composition of 25~50 μ g/ml has the trend of the MCP-1mRNA expression that reduces damage CMEC, and the present composition of 100 μ g/ml reduces the expression of MCP-1.The present composition discharges the trend that IL-8 has reduction to TNF-α damage CMEC.Genipin does not have this effect, and peoniflorin 100 μ g/ml have faint effect, but is weaker than the present composition 25 μ g/ml slightly.
This test shows that the present composition can suppress the effect that TNF-α induces CMEC secretion IL-6, MCP-1, reduces its mRNA and expresses, and prevents the inflammation cascade reaction, alleviates the damage of inflammatory factor to CMEC.The genipin that gives does not separately have this effect, and peoniflorin high concentration (heavy dose) has faint effect.Can draw genipin and Radix Paeoniae and share conclusion with collaborative antiinflammatory action.
3. to the influence of normal rat blood sugar and serum insulin
Method:
8 the week age SD rat random packet, 8 every group.
Each organizes continuous gastric infusion 8d, and every day 1 time, blank group gives the equivalent distilled water.Pluck eyeball in 2 hours after the last administration and get blood, measure blood glucose and serum insulin.
Experiment medicine: glibenclamide group: 25mg/kg;
Genipin group (mg/kg): genipin 120,50;
Radix Paeoniae A organizes (mg/kg): Radix Paeoniae Alba extract (Radix Paeoniae Alba total glycosides content 60%) 12,50;
Radix Paeoniae B organizes (mg/kg): peoniflorin 80,20;
Combination A group (mg/kg): (genipin: Radix Paeoniae Alba extract (Radix Paeoniae Alba total glycosides content 60%)=1: 1) 200,100,50;
Combination B group (mg/kg): (genipin: peoniflorin=1: 2) 80,20;
The result:
Table 3 pair normal rat blood sugar and serum insulin influence (x ± s)
*: with the contrast of blank group, front and back difference P<0.5;
*: with the contrast of blank group, front and back difference P<0.01
Experiment shows: the glibenclamide group can reduce normal rat blood glucose and improve serum insulin levels, and difference has significance.Though the heavy dose of group of genipin can reduce normal rat blood glucose, compares with the blank group, difference does not have significance.Heavy dose of group can improve the serum insulin value, and difference has significance.Each dosage group of peoniflorin and Radix Paeoniae Alba extract group does not reduce normal rat blood glucose, and difference does not have significance, and each dosage group does not have the significance influence to serum insulin.
Each dosage group of the present composition all can reduce the rat euglycemia, but there was no significant difference influences there was no significant difference too to serum insulin.
4. to the influence of blood glucose in diabetic rats due to the chain assistant rhzomorph and serum insulin
Method:
Get male SD rat and prepare low dose of chain assistant rhzomorph by standard method and cause diabetes rat model, behind the 3d according to rat blood sugar value random packet, 8 every group.
Each organizes continuous gastric infusion 8d, and every day 1 time, blank group gives the equivalent distilled water.2 hours (metformin group 1 hour) plucked eyeball and got blood after the last administration, measures blood glucose and serum insulin.
Experiment medicine: positive drug metformin group: 300mg/kg;
Genipin (mg/kg): genipin 100,50;
Radix Paeoniae A organizes (mg/kg): Radix Paeoniae Alba extract (Radix Paeoniae Alba total glycosides content 50%) 400,100;
Radix Paeoniae B organizes (mg/kg): peoniflorin 80,20;
Combination A group (mg/kg): [genipin: Radix Paeoniae Alba extract (paeoniflorin content 60%)=1: 6] 200,100,50;
Combination B group (mg/kg): [genipin: peoniflorin=6: 1] 100,50,30;
The result:
Rat diabetes blood glucose and serum insulin influence due to the table 4 pair chain assistant rhzomorph (x ± s)
*: with the contrast of blank group, front and back difference P<0.5;
*: with the contrast of blank group, front and back difference P<0.01
Experiment shows: the metformin group can effectively reduce this rat model blood glucose, and difference has significance, but does not improve the rat blood serum insulin level.The heavy dose of group of genipin has hypoglycemic activity, and serum insulin is raise the significance influence.All the other dosage groups do not have obvious blood sugar reducing function.
The heavy dose of group of peoniflorin and Radix Paeoniae Alba extract group can reduce rat blood sugar, and difference has significance.Each dosage group does not have the significance influence to serum insulin.
Each group of the present composition all can reduce rat blood sugar, and its effect is better than metformin or suitable with it, and middle dosage (containing) can improve serum insulin for above group.
5. the influence of insulin resistant blood glucose in diabetic rats and serum insulin due to high fat being raised
Method:
Get the male SD rat random packet, 8 every group.
Normal group gives common raising, and all the other each groups give high fat and raise to 8 ages in week, and the 9th week began except that normal group, and all the other give continuous gastric infusion 8d, and every day 1 time, blank group gives the equivalent distilled water.Pluck eyeball in 2 hours after the last administration and get blood, measure blood glucose and serum insulin.
Experiment medicine: positive drug rosiglitazone group: 3mg/kg;
Genipin (mg/kg): genipin 180,80,40;
Radix Paeoniae group (mg/kg): Radix Paeoniae Alba extract (Radix Paeoniae Alba total glycosides content 75%) 360,180,80;
Combination A group (mg/kg): [genipin: Radix Paeoniae Alba extract (Radix Paeoniae Alba total glycosides content 40%)=1: 8] 200,100,50
Combination B group (mg/kg): [genipin: peoniflorin=8: 1] 150,60,30;
The result:
Rat diabetes blood glucose and serum insulin influence due to table 5 pair high fat is raised (x ± s)
*: with the contrast of blank group, front and back difference P<0.5;
*: with the contrast of blank group, front and back difference P<0.01
Experiment shows: the rosiglitazone group has significant difference to rat blood sugar reduction effect, and serum insulin levels is reduced appreciable impact.The genipin heavy dose all has hypoglycemic activity for above group, and serum insulin is not had the significance influence.Dosage all can reduce rat blood sugar for above group in the Radix Paeoniae group, and difference has significance, and reduction has the significance influence to serum insulin.
Each dosage group of the present composition all can reduce rat blood sugar and serum insulin is reduced appreciable impact, and its effect is suitable with rosiglitazone.
Discuss:
Above-mentioned experiment shows, genipin and Radix Paeoniae have the hypoglycemic activity of different mechanism of action for diabetes rat.The genipin mechanism of action is stimulating pancreas β emiocytosis insulin, and Radix Paeoniae acts on and reduces insulin resistant and may urge the periphery glucose utilization.
The present composition has above-mentioned all mechanism of action simultaneously, and greatly reduce single pharmaceutical quantities, and advantageously share of the two do not reduced euglycemia, and wherein mechanism is at present theoretically still can't explain fully, infer its mechanism may be for receptor mechanism behind the saturated and receptor.