CN104042613B - A kind of composition and the purposes in medicine thereof - Google Patents
A kind of composition and the purposes in medicine thereof Download PDFInfo
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- CN104042613B CN104042613B CN201410260301.8A CN201410260301A CN104042613B CN 104042613 B CN104042613 B CN 104042613B CN 201410260301 A CN201410260301 A CN 201410260301A CN 104042613 B CN104042613 B CN 104042613B
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Abstract
The present invention relates to a kind ofly mix by Rhein, jamaicin the composition forming, for prevention and treatment diabetes, ephrosis, belong to field of medicaments. A kind of composition, is characterized in that example meter in mass ratio, Rhein: jamaicin is 1: 2-1: 10. The present invention shows that by great many of experiments Rhein and jamaicin are 1 at mass ratio: 2-1: in 10 scopes, synergistic function occurs, the side effects such as the diarrhoea of Rhein do not appear in discovery simultaneously. We find not (as comparative example 1 or 2) not synergy in proportion of the present invention simultaneously, therefore illustrate that special ratios of the present invention is the place of key of the present invention.
Description
Technical field
The present invention relates to a kind ofly mix by Rhein, jamaicin the composition forming, for prevention and treatment diabetes, kidneyDisease, belongs to field of medicaments.
Background technology
Rhein, chemical name 1,8-Dihydroxy-3-carboxy-anthraquinone, molecular formula is C15H8O6, be coffee-like needle, after distillationFor yellow needle, be not dissolved in water, be slightly soluble in ethanol, ether, benzene, acetone. In medicinal herbs most in use rheum officinale, folium sennae, the fleece-flower rootAll contain Rhein, be used for the treatment of clinically the disease such as diabetic nephropathy, diabetes. But Rhein easily causes diarrhoea, cause and rush downReason may be that the metabolite anthrone (anthrone) in enteric microorganism system causes by it. In addition, the dissolving of RheinProperty is also poor, the water insoluble and alcohol of Rhein, and be insoluble to most organic solvents, bioavilability is lower. Two secondAlthough acyl Rhein can pass through oral administration, can not be absorbed by intestines and stomach completely. This makes the clinical use of RheinBe restricted.
Jamaicin is the known antibacterials of commonly using, and molecular formula is C20H16ClNO4, yellow crystalline powder, odorless, tasteExtremely bitter, mainly come from the coptis, barberry, jamaicin molecular weight is 480. Jamaicin can be separated out yellow needle-like crystalline substance from etherBody. 145 DEG C of fusing points, water-soluble, be insoluble in benzene, EC. Jamaicin is a kind of quartermary ammonium alkaloids, and its esters is in waterSolubility is all smaller, and for example hydrochloride is 1: 500, and sulfate is 1: 30. Maximum feature is that side effect is extremely rare, clinicalOn be used for the treatment of bacillary dysentery and enteritis, be also useful on treatment diabetes aspect report. But jamaicin exists solubility low(being only 0.2%), body absorb the problems such as difficulty, difficulty are transported in diseased region and cell effectively, cause its biological profitExpenditure low (being generally 10%, < 25%), causes patient's medication number of times frequent, and tolerance is poor, and drug effect is poor, greatly limits that it is clinicalApplication.
Inventor finds under study for action, and jamaicin, Rhein have the effect of mutually promoting and dissolving, and hint can be carried mutuallyHigh bioavilability, and further confirmed this effect by pharmacological evaluation, reduced the toxic and side effect of Rhein simultaneously. OrderBefore there is no the report that two kinds of Rheins, jamaicin mix the composition of composition, there is no said composition for prevention and treatment sugar yetUrine is sick, the report of ephrosis.
Summary of the invention
The invention discloses a kind of composition, said composition is mixed and is formed by Rhein, jamaicin; The invention also disclosesThe medical usage of said composition in the prevention from suffering from the diseases such as diabetes, ephrosis and treatment.
A kind of composition, is characterized in that example meter in mass ratio, Rhein: jamaicin is 1: 2-1: 10.
Described a kind of composition, is characterized in that example meter in mass ratio, Rhein: jamaicin 1: 5.
Described composition is in preparation prevention and the application for the treatment of in diabetes medicament.
Described composition is in preparation prevention and the application for the treatment of in ephrosis medicine.
Rhein described in the present invention also comprises the compound that salt that itself and alkalinous metal form or various salt form;Described jamaicin band or the not compound with the crystallization water or the formation of the different crystallization water.
Composition in the present invention can be used as the purposes of unique composition for medical aspect, also can add that other are effectiveComposition; Be used for the treatment of 1 type and diabetes B, diabetic nephropathy, glomerulonephritis, chronic mesentery nephritis, chronic film kidneyScorching.
Composition in the present invention is oral and injection shape for prevention and treatment diabetes, ephrosis medicine and health productsFormula; Concrete form is tablet, capsule, granule, injection with small volume, high-capacity injection, freeze-dried powder.
Beneficial effect
The present invention shows that by great many of experiments Rhein and jamaicin are 1 at mass ratio: 2-1: in 10 scopes, occur collaborativeSynergistic effect, there is not the side effects such as the diarrhoea of Rhein in discovery simultaneously. We find not at ratio model of the present invention simultaneouslyIn enclosing, (as comparative example 1 or 2) not synergy, therefore illustrates that special ratios of the present invention is the place of key of the present invention.
Brief description of the drawings
Fig. 1 Rhein and Berberine hydrochloride animal vivo test blood concentration-time graph
Detailed description of the invention
Embodiment 1
Rhein, jamaicin, starch, microcrystalline cellulose, PVP K30 are put in Quick-stirring granulator and are mixed,Add purified water wetting softwood processed in right amount, cross 18～24 orders and granulate, 40～70 DEG C are dried to loss on drying and are less than 6%, mistake 24～The whole grain of 60 orders, adds dolomol, mixes, and particle is compressing tablet after the assay was approved.
Embodiment 2
Rhein, jamaicin, starch, microcrystalline cellulose, PVP K30 are put in Quick-stirring granulator and are mixed,Add purified water wetting softwood processed in right amount, cross 18～24 orders and granulate, 40～70 DEG C are dried to loss on drying and are less than 6%, mistake 24～The whole grain of 60 orders, adds dolomol, mixes, and particle is compressing tablet after the assay was approved.
Embodiment 3
Rhein, jamaicin, starch, microcrystalline cellulose, PVP K30 are put in Quick-stirring granulator and are mixed,Add purified water wetting softwood processed in right amount, cross 18～24 orders and granulate, 40～70 DEG C are dried to loss on drying and are less than 6%, mistake 24～The whole grain of 60 orders, adds dolomol, mixes, and particle is compressing tablet after the assay was approved.
Comparative example 1
Rhein, jamaicin, starch, microcrystalline cellulose, PVP K30 are put in Quick-stirring granulator and are mixed,Add purified water wetting softwood processed in right amount, cross 18～24 orders and granulate, 40～70 DEG C are dried to loss on drying and are less than 6%, mistake 24～The whole grain of 60 orders, adds dolomol, mixes, and particle is compressing tablet after the assay was approved.
Comparative example 2
Rhein, jamaicin, starch, microcrystalline cellulose, PVP K30 are put in Quick-stirring granulator and are mixed,Add purified water wetting softwood processed in right amount, cross 18～24 orders and granulate, 40～70 DEG C are dried to loss on drying and are less than 6%, mistake 24～The whole grain of 60 orders, adds dolomol, mixes, and particle is compressing tablet after the assay was approved.
Embodiment 4
1, composition affects STZ blood glucose in diabetic rats
Material:
Composition makes by embodiment 1, embodiment 2, embodiment 3, comparative example 1, comparative example 2 respectively; STZ (Sigma public affairsDepartment); SD rat 200 ± 20g (Guangdong Medical Lab Animal Center). Method:
200 of SD rats, male and female half and half, get 180 every lumbar injection 10mgkg-1STZ measured blood after 72 hoursSugar concentration, selected fasting blood-glucose is at 10mmolL-1100 of above animals, then by being divided at random 8 groups, 10 every group; RespectivelyFor model group, embodiment 1, embodiment 2, embodiment 3, Rhein group, jamaicin group, comparative example 1 and comparative example 2, remain 20Animal is blank group.
Result
Table 1 gastric infusion 40 days is to diabetes rat fasting blood-glucose (mmolL due to STZ-1) impact
Note: with model group comparison,*P＜0.05，**P＜0.01
Result shows: after medicine 40 days, and embodiment 1, embodiment 2 and embodiment 3, after administration, 40 days blood sugar levels are all lower than mouldType group (P < 0.01), is better than independent medication Rhein group and jamaicin group, and hypoglycemic effect is about more than 2 times, approaches blank group and fallsSugar effect is remarkable, does not find rat diarrhoea problem simultaneously. But compare lower than model group (P < with model group after the administration of Rhein group0.05), there is rat diarrhoea phenomenon simultaneously. Comparative example 1 and comparative example 2 differ not with using separately jamaicin and Rhein groupGreatly, lower than model group (P < 0.05).
Illustrate: embodiment of the present invention 1-3 has synergistic function, avoided the side effect of Rhein simultaneously. Also explanationComparative example 1 and 2, not in proportion of the present invention, does not have synergistic effect.
2, composition on STZ diabetes rat glucose load after the impact of blood sugar
By the dosage successive administration of above-mentioned test 2 days, rat fasting administration after 3 hours in the 3rd day, respectively organize rat respectively toAfter medicine 15min, gavage gives 2.5gkg-1Glucose, and giving after glucose 0,0.5,1,2h, survey the empty stomach after glucose loadChange of blood sugar.
Table 2 gastric infusion 3 days is to blood glucose in diabetic rats (mmolL due to STZ-1) impact
Note: with model group comparison,*P＜0.05，**P＜0.01
Result shows: embodiment 1-3 can significantly reduce blood glucose in diabetic rats due to the STZ of glucose load compared with model groupLevel (P < 0.01), and comparative example 1-2 with separately with Rhein group with jamaicin group compared with model group, can reduce loadBlood glucose in diabetic rats level due to STZ (P < 0.05). Find that Rhein group has diarrhoea phenomenon simultaneously. Illustrate after administration, implementExample 1-3 is than using separately Rhein or jamaicin, or comparative example 1 and 2 effects are all good, has reduced secondary work of poison of Rhein simultaneouslyWith. Comparative example 1 and 2 is also described not within the scope of the present invention, there is no synergistic effect.
2, the impact of composition on diabetes B opposing rat blood serum phylaxin
Material:
Composition is pressed respectively embodiment 3, and comparative example 1 and comparative example 2 make; (Guangdong Province's medical science is real for SD rat 200 ± 20gTest animal center); Insulin radioimmunoassay kit (Beijing Fu Rui bioengineering Co., Ltd); Resistin detects examinationAgent box (PhoenixBiotech company of the U.S.).
Method
Except 20 animals of blank, all the other 80 animals feed and raise high fat diet, and within the 10th week, animal pattern is pressed 25mgkg-1Disposable celiac STZ, is divided into 4 layers by animal pattern by body weight, then is randomized into 4 groups by body weight, and 20 every group is modelGroup, embodiment 3, comparative example 1, comparative example 2, another 20 is blank group, successive administration 30 days is got blood and is surveyed serum on an empty stomach on socket of the eyeInsulin and Serum Resistin Levels.
Result
Table 3 is to rat blood sugar level (mmolL-1) impact
Note: with model group comparison,*P＜0.05，**P＜0.01
Result shows: comparative example 1 and 2 administrations, after 30 days, can reduce rat blood sugar level (P < 0.05). But embodiment 3 utmost pointsSignificantly reduce rat blood sugar level (P < 0.01).
Table 4 composition is to rat limosis insulin (mUL-1) impact
Note: with model group comparison,*P＜0.05，**P＜0.01
Result shows: comparative example 1 and 2 administrations, after 30 days, can obviously reduce rat blood serum insulin level (P < 0.05);Embodiment 3 is (P < 0.01) particularly significantly.
The impact of table 5 composition on rat ISI (insulin sensitivity index)
Note: with model group comparison,*P＜0.05，**P＜0.01
Result shows: embodiment 3 administrations, after 30 days, can obviously improve rat insulin Sensitivity Index (P < 0.05). And it is rightRatio 1 and 2 there was no significant differences.
Impact (the μ gL of table 6 composition on rat blood serum phylaxin-1)
Note: with model group comparison,*P＜0.05，**P＜0.01
Result shows: embodiment 3 administrations, after 30 days, can obviously reduce rat blood serum resistin concentration (P < 0.05).
And comparative example 1 and 2 there was no significant differences.
4, the impact of composition on type 1 diabetes Renal Function in Rats
Material:
Composition is pressed respectively embodiment 1, and comparative example 1 and comparative example 2 make; SD rat is (in Guangdong Province medical experiment animalThe heart).
Method:
Adult male SD rats, weight 180-220g. Adaptability is fed after 7d. After 12h, press random packet: modeling group 50Only give lumbar injection STZ60mg/kg (being dissolved in 0.1mmol/I citric acid one sodium citrate buffer solution of pH4.5); Normal controlOrganize 10 and give single intraperitoneal injection same volume 0.1mmol/L citrate buffer solution. After 72h, tail venous blood sampling is surveyed random blood sugar.With blood sugar) 16.7mmol/L, be defined as diabetes model. Diabetes become mould rat to be divided at random 5 groups. Every group 10: diabetesNon-treatment group, comparative example 1 are pressed 40mgkg-1·d-1Administration, comparative example 2 are pressed 40mgkg-1·d-1Administration, embodiment 1 press40mg·kg-1·d-1Administration. Adaptability was raised after 1 week, gave to process accordingly factor by grouping.
Continue medication 12 weeks, claim weight, stay 24h urine with metabolic cage, note urine amount, surveys 24h urine albumen (UPRO24), urineCreatinine (UCR), microdose urine protein (UMA), heart extracting blood after 10% chloraldurate solution 4.5ml/kg anesthesia, 3000r/minCentrifugal, separation of serum ,-20 DEG C of preservations. survey blood urea nitrogen (BUN), serum creatinine (Scr). CrCl Ccr presses formula: urine fleshAcid anhydride concentration (mg/L) × per minute urine amount (mg)/serum creatinine concentration (mg/ml) is calculated. and proofread and correct with weight. Continue medication 12 weeksRear execution, gets right kidney.
Result
The ordinary circumstance SD rat of rat is after lumbar injection STZ, and significantly many drinks, many all appear in the non-treatment group of diabetesFood, diuresis and the disorderly symptom of the diabetic supersession such as become thin, chaeta is in disorder, slow in reacting, and activity obviously reduces, and has rat to go outThe symptoms such as existing cataract, infection and skin ulceration. The general state of the each treatment group rat of diabetes all more non-treatment group changes to some extentKind. wherein better with dosage group in composition, high dose group group rat state. Include 60 of rats in, all enter interpretation of result. nothingDe-mistake.
The impact of table 7 composition on diabetes rat renal function
*With relatively P < 0.05 of Normal group;#With relatively P < 0.05 of the non-administration group of diabetes
The impact of table 8 composition on diabetes rat kidney quality/weight
*With relatively P < 0.05 of Normal group;#With relatively P < 0.05 of the non-administration group of diabetes
Result shows: diabetes become module rat BUN, Ccr, UPRO24, the equal compared with normal group of UMA (the P < that obviously raises0.05). The each treatment group BUN of diabetes, Ccr, UMA all more non-treatment group obviously reduce. Diabetes modeling is respectively organized renal hypertrophy and is referred toNumber compared with normal control group obviously increases (P < 0.05). Each treating diabetes group renal hypertrophy index is compared with under the non-treatment group of diabetesFall. But embodiment 1 effect is better than comparative example 1 and 2.
5, composition causes the therapeutic action that improves of chronic mesangial proliferative glomerulonephritis to monoclonal antibody 1-22-3
Material:
By comparative example 1, comparative example 2 and embodiment 2 make respectively; Wistar rat is (in Guangdong Province medical experiment animalThe heart).
Method
In 7 week age of 30 male Wistar rats, divide at random three groups, 10 every group. The 1st group: oppose to physiological saline (PBS)According to; 2, the four groups of embodiment 2 of 1, the 3 group of comparative example of the 2nd group of comparative example. Excise left kidney after 24 hours, 500 μ g are mono-for tail vein injectionClonal antibody 1-22-3 (purchased from Panafarm experiment Co., Ltd) gastric infusion simultaneously, observes 42 days altogether. Within the 42nd day, put to death completePortion rat, heart extracting blood, after winning right kidney and weighing, gets nephridial tissue and does immunofluorescent.
Result
1. urine albumen result:
Table 924 hour urine albumen result
Note: compare with PBS group*P＜0.05，**P＜0.001
Result shows: embodiment 1 is compared with comparative example 1 and 2 better effects if.
2. immunofluorescence result:
Embodiment 2 can alleviate the pathological lesion of glomerulus. Our semi-quantitative analysis composition transforming growth factor β(TGF-β), the impact of type i collagen (CollI) and rat smooth muscle actin (α-SM-actin) etc., we find medicineAfter treatment, suppressed significantly the expression in glomerulus such as TGF-β, CollI and α-SM-actin, statistical procedures has significantlyThe difference (in table 6) of property.
Table 10 immunofluorescence semi-quantitative analysis result
Note: compare with PBS group***P＜0.001
3. the impact on blood biochemistry index:
Table 11 blood biochemistry index
Note: compare with PBS group,*P＜0.05，**P＜0.01
Embodiment 2 conspicuousness compared with PBS reduces T-chlest, and BUN, is better than comparative example 1 and 2.
Embodiment 5
Solubility test
Rhein has the carbonyl structure of exposure, and jamaicin has the nitrogen-atoms of exposure, and the two approaches likely and formsShare electron pair, carries out combination by intermolecular force. We have carried out solubility test for this reason, and we have carried out 12 groups altogetherExperiment, the amount of every group of experiment reservation Rhein is constant, increases progressively at double gradually the dosage of jamaicin, required thereby observation is all dissolvedThe consumption of water, tests parallel three times, averages for every group. Concrete data see the following form 12
Table 12 Rhein and Berberine hydrochloride solubility experiment
By experiment, we find, Berberine hydrochloride can obviously increase the solubility of Rhein in water. Point out the twoShare and may there is synergy.
Pharmacokinetics experiment
For proving Rhein of the present invention and Berberine hydrochloride bioavilability in vivo, taking Rhein as reference medicineThing, carries out bioavilability and evaluation of bioequivalence. 18 SD rats, male and female half and half, weight range 200～250g, experimentFasting the previous day 12h, unified feed low fat standard meal after administration 4h. Must not take other at test the last fortnight and duration of testMedicine. 18 SD are divided into 3 liang of groups (6 every group) at random. Press respectively Rhein (140mg/kg), Rhein: Berberine hydrochloride(140mg/kg: 560mg/kg, 1: 4), Rhein: Berberine hydrochloride (70mg/kg: 280mg/kg, 1: 4) oral dose administration,Before taking medicine, get blank blood, the 0.5mL that regularly takes a blood sample after taking medicine, in scribbling the test tube of heparin, centrifugal immediately, get upper plasma, coldFreeze preservation. Add acetonitrile 5ml, mix 5 minutes, centrifugal 30 minutes of 5000rpm, gets supernatant 2ml, and nitrogen dries up, and freezing preservation is measuredFront with sample introduction after mobile phase dissolving, measure the vivo medicine concentration of Rhein by HPLC method.
Pharmacokinetic parameter in table 13 Rhein and Berberine hydrochloride animal body
Found that Rhein: after Berberine hydrochloride (1: 4) administration, Rhein is pressed 70mg/kg administration and alone in vivoThe Cmax of Rhein 140mg/kg is suitable, AUC0-∞Similar to alone Rhein, Tmax and t1/2Unchanged, Rhein is described: saltThe oral Rhein AUC of acid jamaicin (1: 4) is significantly increased, and Rhein and Berberine hydrochloride share and can improve Rhein mouthThe bioavilability of clothes; And Rhein: Tmax and AUC that Berberine hydrochloride (1: 4) share0-∞There is dose dependent, with administrationDosage becomes positive correlation. The results are shown in accompanying drawing 1 and table 13 and show, this has great advantage at clinical tool.
Claims (6)
1. a pharmaceutical composition, is characterized in that example meter in mass ratio, Rhein: jamaicin is 1:4 ~ 10.
2. a kind of composition according to claim 1, is characterized in that counting in mass ratio Rhein: jamaicin is 1:5.
3. according to claim 1-2 any one composition, in described composition, Rhein, jamaicin are prototype or pharmacyAcceptable salt.
4. composition according to claim 1 is in preparation prevention and the application for the treatment of in diabetes medicament.
5. composition according to claim 1 is in preparation prevention and the application for the treatment of in ephrosis medicine.
6. composition according to claim 5 is in preparation prevention and the application for the treatment of in medicine for treating diabetic nephropathy.
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