CN100486567C - Curcumin emulsion and its preparation process - Google Patents

The invention provides a curcumin oral emulsion and injection, wherein the emulsion comprises curcumin, oil, emulsifying agent and water, the injection also contains isotonic agent. The invention also discloses the process for preparing the emulsion and its use in preparing medicaments for treating or preventing tumor, resisting tumor transfer, enhancing curative effects of chemotherapy drugs, reducing side effects of chemotherapy and improving immunity functions for tumor patients.

Curcumin emulsion and preparation method thereof

Invention field

The present invention relates to curcumin emulsion and its production and use.

Background technology

Generalized curcumin is commonly referred to curcumin, be by Curcuma conventional Chinese medicine Rhizoma Curcumae Longae (Curcuma long L.), Radix Curcumae (Curcuma wenyujin Y.H.Chen et C.Ling), the effective ingredient of separation and Extraction in the dried root of Guangxi zedoary (Curcuma kwangsiensis S.G.Leeet C.F.Liang) or Rhizoma Curcumae (Curcuma phaeocaulis Val.), comprise curcumin (curcumin), demethoxycurcumin (demethoxycurcumin) and bisdemethoxycurcumin (bis-demethoxycurcumin) [Liu Shuxing, Hu Xiaojun, Shaanxi Tech Univ's journal: natural science edition, 2003].When below not doing specified otherwise, curcumin promptly refers to generalized curcumin, and their structural formulas are as follows:

I curcumin Curcumin R ₁=R ₂=CH ₃O

II demethoxycurcumin Demethoxycurcumin R ₁=H, R ₂=CH ₃O

III bisdemethoxycurcumin Bisdemethoxycurcumin R ₁=R ₂=H

Curcumin is water insoluble, and oral administration biaavailability is very low, most of with original shape excrete (about 89%) [Liu Anchang, Lou Hongxiang, Zhao Lixia. curcumin pharmacologically active and internal metabolism. external medical plant amedica fascicle .2004; 19 (1): 1-5].

The solution instability of curcumin has generally speaking been described in [Chinese Pharmacological circular, 1998,14 (5): 415-417] and curcumin has been dissolved in the D-hanks solution after making cosolvent with DMSO, is placed at 4 ℃ that external antitumor activity promptly obviously descends in 3 days.Described the hydrotropy with DMSO in [Chinese crude drug, 2002,8 (25): 585-586], with the dissolving of Hanks liquid, the effect duration of the curcumin injection of making is 0.2 year, and draws the conclusion that is not suitable for curcumin is developed to injection.

In view of the foregoing, Orally taken emulsion that the preparation bioavailability is high and stable and the stable curcumin injectable emulsion of preparation are that people thirst for the difficult problem that solves for a long time always.

Summary of the invention

The invention provides curcumin emulsion, it can be that oral breast also can be the injection breast.

The invention provides the preparation method of curcumin emulsion.

The invention provides curcumin emulsion at preparation treatment or prophylaxis of tumours, anti metastasis, enhancing chemotherapeutics therapeutic effect, alleviate chemical therapy toxic side effect and improve application aspect the tumor patient immunologic function.

Curcumin emulsion provided by the invention contains curcumin, oil, emulsifying agent and water, and in Emulsion weight, the curcumin consumption is 0.0001-5%, and oily consumption is 1-40%, and the emulsifying agent consumption is 0.1-10%, and water consumption is 50-98%.

When curcumin emulsion provided by the invention existed with injection type, it also contained the accent isotonic agent, and transferring isotonic agent is glycerol, and its consumption is 0.1-5%.

Emulsion provided by the invention, oil can be one or more in vegetable oil or the quintessence oil, and vegetable oil is selected from soybean oil, Semen Maydis oil, Oleum Arachidis hypogaeae semen, olive oil, Oleum Ricini, Petiolus Trachycarpi oil, Oleum Cocois, Oleum Brassicae campestris, Oleum sesami, medium-chain fatty acid or Oleum Gossypii semen; Quintessence oil is selected from Fructus Schisandrae oil, Oleum Hippophae, peach kernel oil, almond oil, safflower oil, Oleum Camelliae, Radix Oenotherae erythrosepalae oil, Semen Tritici aestivi germ oil, perilla oil, litsea cubeba oil, Oleum Fructus Bruceae or Oleum Curcumae; Be preferably be rich in chain fatty acid triglycerides and MCT Oil Oleum Glycines.

Emulsion provided by the invention, emulsifying agent can be one or more in amphoteric surfactant, the non-ionic surface active agent.Wherein amphoteric surfactant is selected from one or more in fabaceous lecithin, egg yolk lecithin, phosphatidylcholine, PHOSPHATIDYL ETHANOLAMINE, serinephosphatide, lipositol, phosphatidic acid, cephalin, the hydrogenated phospholipid; Non-ionic surface active agent is selected from one or more in sucrose-fatty esters (as sucrose ester), spans, Tweens, polyethylene pyrrole Lip river alkane ketone, polyoxyethylene fatty acid ester class (as Myrij Myrij), polyoxyethylene aliphatic alcohol ether class (as Bian Ze Brij), Cetomacrogol class, Cremophore EL class or the poloxamer class (as poloxamer).Be preferably in soybean phospholipid, egg yolk lecithin, hydrogenated phospholipid, the polyoxyethylene polyoxypropylene block polymer (poloxamer) one or more.

Emulsion provided by the invention can also contain additive, and additive can be the pH regulator agent, as sodium hydroxide, hydrochloric acid; Can be glycerol also, consumption be about 0-5%, can also be sterol, and as cholesterol or long-chain (C14-C22) alcohol, oleic acid, linoleic acid and derivant, cyclodextrin and derivant thereof, its consumption is 0-10%.

The preparation method of curcumin emulsion of the present invention is: after making the curcumin oil solution, with emulsifying agent this solution is distributed to and makes stabilized oil-in-water emulsion in the water.Be specially: curcumin is dissolved in the alcoholic solution, additive, emulsifying agent can be joined in this alcoholic solution, be added to this solution in the oil and be mixed to clear and bright; Remove alcohol with rotary evaporation or the evaporation of logical nitrogen current; Emulsifying agent is distributed in the water, whenever additive glycerol can be added to aqueous phase in preparation process; The oil solution that contains curcumin following of high-speed stirred is added to aqueous phase and forms thick Emulsion (but direct packaging becomes Orally taken emulsion); The thick Emulsion of gained must be suitable for injecting stable, even, the nontoxic Emulsion of use by ultrasonic disintegrator, the further emulsifying of high pressure dispersing emulsification machine.

Orally taken emulsion provided by the invention has solved the low defective of oral administration biaavailability.The inventor provides the curcumin injectable emulsion, has solved the technical barrier that people can't make curcumin injection for a long time.Emulsion curcumin provided by the invention is wrapped in the oil or is wrapped in the micelle of emulsifying agent, reduces the contact that reaches external environment with aqueous dispersion mutually, has obviously improved the stability of curcumin.Said preparation has also obviously improved curcumin antineoplastic therapeutic effect, suppresses the transfer of tumor cell more effectively.During clinical use, biodegradable emulsifying agent and provide the oil phase of energy can improve patient's life quality in the Emulsion increases immunologic function.This Emulsion is stablized, can be tolerated autoclaving, and the adjuvant of use has biocompatibility, and shows less side effect.The maximum tolerated dose experiment shows that mouse tail vein gives curcumin emulsion 500mg/kg, does not observe obvious toxic and side effects.Emulsion provided by the invention is efficient, low toxicity, is suitable for commercial production.

The specific embodiment

Preparation example 1: preparation curcumin Orally taken emulsion

Curcumin 50mg, soybean phospholipid 2.0g, cholesterol 0.5g are joined among the refined soybean oil 10g after with dissolve with ethanol, be mixed to clear and bright, and by rotary evaporation or feed vaporized nitrogen and remove alcohol, obtain containing the oil solution of curcumin 5mg/g, fabaceous lecithin 200mg/g and 50mg/g cholesterol.

Poloxamer 1.0g is dispersed in an amount of water for injection, and under the high-speed stirred condition, the oil solution that slowly drips the phospholipid contain curcumin, cholesterol is made thick Emulsion in aqueous dispersion, and thin up is to 100ml.Emulsifying finishes the back in 100 ℃, the 30min sterilization, and packing promptly gets the Emulsion that contains curcumin 0.5mg/ml.

Preparation example 2: preparation curcumin injectable emulsion

Curcumin 500mg, oleic acid 500mg are joined among the refined soybean oil 10g after with dissolve with ethanol, be mixed to clear and brightly, and by rotary evaporation or feed vaporized nitrogen and remove alcohol, obtain containing the oleic soybean oil solution of curcumin 50mg/g and 50mg/g.

With poloxamer 3.0g, glycerol 2.25g is added in an amount of water for injection, under the high-speed stirred condition, slowly drips and contains curcumin, oleic oil solution in above-mentioned aqueous dispersion, makes thick Emulsion, is diluted to 100ml with water for injection.Under the condition of pressure 60Mpa, by the further emulsifying of dispersing emulsification machine, emulsifying finishes the back in 100 ℃ with resulting thick Emulsion, the 30min sterilization, and packing promptly gets injection curcumin emulsion 5mg/ml.

Preparation example 3: preparation curcumin Orally taken emulsion

Curcumin 5000mg, egg yolk lecithin 3.0g are joined among the Semen Maydis oil 40g after with dissolve with ethanol, be mixed to clear and brightly, and remove alcohol, obtain containing the Semen Maydis oil solution of curcumin 125mg/g, egg yolk lecithin 75mg/g by rotary evaporation.

Under the high-speed stirred condition, the oil solution that slowly drips the phospholipid that contains curcumin adds in the aqueous solution of an amount of 1% hydroxypropyl beta cyclodextrin, makes thick Emulsion, adds pure water to 100ml, and packing promptly gets curcumin emulsion 50mg/ml.

Preparation example 4: preparation curcumin injectable emulsion

Curcumin 200mg, hydrolecithin 2.0g are joined among the soybean oil 10g after with dissolve with ethanol, be mixed to clear and brightly, and remove alcohol, obtain containing the soybean oil solution of curcumin 20mg/g, hydrolecithin 200mg/g by rotary evaporation.

Glycerol 2.25g is added to an amount of water for injection, under the high-speed stirred condition, slowly drips the oil solution that contains curcumin and hydrolecithin and add the above-mentioned aqueous phase that arrives, make thick Emulsion, be diluted to 100ml with water for injection.Under the condition of pressure 60Mpa, by the further emulsifying of dispersing emulsification machine, emulsifying finishes the back in 115 ℃ with resulting thick Emulsion, the 30min sterilization, and packing promptly gets injection curcumin emulsion 2mg/ml.

Preparation example 5: preparation curcumin Orally taken emulsion

Curcumin 50mg, soybean phospholipid 2.0g are joined among the refined soybean oil 10g after with dissolve with ethanol, be mixed to clear and brightly, and by rotary evaporation or feed vaporized nitrogen and remove alcohol, obtain containing curcumin 5mg/g, fabaceous lecithin 200mg/g oil solution.

Poloxamer 1.0g is dispersed in an amount of water for injection, and under the high-speed stirred condition, slowly the oil solution that drips the phospholipid that contains curcumin is made thick Emulsion in aqueous dispersion, and thin up is to 100ml.Emulsifying finishes the back in 100 ℃, the 30min sterilization, and packing promptly gets the Emulsion that contains curcumin 0.5mg/ml.

With preparation example 4 is example, detects the degraded situation of curcumin emulsion under the different temperatures, detection method: HPLC method

Table 1: the degraded of curcumin in the Emulsion under the different temperatures

By table 1 /relational expression of lnC/ and t can try to achieve the degradation rate constant k under the relevant temperature.See Table 2

Table 2: the degradation rate constant of curcumin emulsion under the different temperatures

With lnk 1 * 1000/T linear regression is got regression equation,

lnk＝-10.83*×(1×1000/T)+24.937 r＝0.9835

Activation energy Ea=90.07kJ/mol

k _25℃＝1.098×10 ^-5d ^-1，t _0.9＝399.8d

According to calculating, curcumin emulsion Chinese medicine content drops to the former 90% required time of drug level and is about 400 days.

With preparation example 3 is example, with curcumin emulsion in 30 ℃, placed 3 months under the condition of relative humidity RH75%, indexs such as the particle size distribution of detection Emulsion, outward appearance, pH value, viscosity, content, testing result shows, place under these conditions, each of Emulsion detects index mutually and there is no significant change, illustrates that the physicochemical property of Emulsion is more stable under these conditions.The results are shown in Table 3.

The accelerated test of table 3 curcumin emulsion is investigated

The tumor-inhibiting action of experimental example 1 curcumin vein emulsion and its injection relatively

Press document [Tan Jun, Cai Zhuofan, Wang Shiming. curcumin preparation stability test research. Chinese crude drug, 2002,8 (25): 585-586] method prepares the curcumin injection, prepares curcumin emulsion by preparation example 2 methods.Sterilization back room temperature is placed.Injection is now with the current, and be no more than 1 month service time after curcumin emulsion prepared.

Kunming mouse is provided by Shandong Province's natural drug Engineering Technical Research Centre animal center, and C57BL/6 pure lines Mus is available from Chinese Academy of Medical Sciences's animal center.H22 rat liver cancer and the strain of Lewis lung cancer tumor are drawn from pharmacological room of Beijing medicine institute.

Get the good H22 ascites of all growth conditions that the goes down to posterity mice of going down to posterity, get ascites after, add the long-pending injection normal saline dilution of triploid, be inoculated in 80 Kunming mouse oxters, every 0.2ml.Be taken at oxter go down to posterity inoculation back the tenth day and the good Lewis lung cancer tumor tissue of growth conditions, add the normal saline of quintuple, cross 100 eye mesh screens after tissue grinder's homogenate, the gained tumor cell suspension is inoculated in 80 C57BL/6 pure lines Mus oxters, and 0.2ml/ only.Connect after the tumor second day, by body weight mice is divided into 8 groups at random respectively, 10 every group.Be respectively model group, positive drug group (cyclophosphamide 20mg/kg), curcumin injection height (50mg/kg), in (25mg/kg), low (12.5mg/kg) dosage group and curcumin emulsion height (50mg/kg), in (25mg/kg), low (12.5mg/kg) dosage group.Every day the tail intravenously administrable once, successive administration 10 days.After the last administration, put to death animal in 24 hours, weigh, strip tumor tissue and weigh, calculate each administration group tumour inhibiting rate.Adopt student t test that curcumin emulsion group and injection group tumor are heavily compared.

The tumor-inhibiting action of table 4. curcumin injection and curcumin emulsion

With same dose injection group tumor anharmonic ratio: ^*, P＜0.05; ^*, P＜0.01.

The results are shown in Table 4, curcumin vein emulsion and injection all are the growth that dose dependent ground has suppressed mice H22 hepatocarcinoma and Lewis lung cancer, but the tumor-inhibiting action of the curcumin emulsion of same dose is better than the injection group, and statistical result shows under height (50mg/kg), the middle dosage (25mg/kg) the significance difference.Illustrate that the curcumin emulsion antitumous effect is better than common curcumin injection under the same dosage.

The influence that experimental example 2 curcumin emulsions shift tumor cell

As experimental example 1 preparation curcumin injection and curcumin emulsion.

80 of C57BL/6 mices, body weight 18 ~ 22g.Each mice is in tail vein injection B16/BL6 tumor cell (available from institute of materia medica, Chinese Academy of Medical Sciences Beijing) suspension 1 * 10 ⁶/ only.Mice was divided into 7 groups at random by body weight in second day, be respectively model group, curcumin injection height (50mg/kg), in (25mg/kg), low (12.5mg/kg) dosage group and curcumin emulsion (50mg/kg), in (25mg/kg), low (12.5mg/kg).Each is organized the tail vein and gives relative medicine, and model group is given the injection normal saline, dosage 0.2ml/10g body weight.Successive administration 21 days.After the last administration 24 hours, disconnected neck was put to death mice, strips the full lung of mice, and anatomical lens is observed branch on count lung tuberosity number down, calculated and respectively organized mouse lung metastasis mean, and utilize student t test to organize a statistics relatively.

Experimental result shows that curcumin emulsion senior middle school low dose group all can obviously suppress the transfer of tumor, and the lung that injection group high dose and middle dosage also can obviously suppress tumor shifts.Under the same dose, Emulsion has stronger metastasis effect than injection.

The influence that table 5 curcumin emulsion and injection shift mouse black-in lymphoma B16/BL6 lung

Compare with model group: ^#, P＜0.05; ^##, P＜0.01.Compare with same dose injection group: ^*, P＜0.05; ^*, P＜0.01.

Experimental example 3 curcumin emulsions are to tumor-bearing mice chemotherapy Attenuation

Prepare curcumin emulsion by preparation example 4 methods.Sterilization back room temperature is placed.Kunming mouse is provided by Shandong Province's natural drug Engineering Technical Research Centre animal center.The strain of H22 Mouse Liver carcinoma is drawn from pharmacological room of Beijing medicine institute.

Get the good H22 ascites of all growth conditions that the goes down to posterity mice of going down to posterity, get ascites after, add the long-pending injection normal saline dilution of triploid, be inoculated in 80 Kunming mouse oxters, every 0.2ml.Connect after the tumor second day, by body weight mice is divided into 8 groups at random respectively, 10 every group.Be respectively model group, chemotherapy group (cyclophosphamide 100mg/kg * 2 time), curcumin group (curcumin 50mg/kg/ day), chemotherapy merges curcumin emulsion height (cyclophosphamide 100mg/kg/5 day, curcumin 50mg/kg/ day), (cyclophosphamide 100mg/kg/5 day in, curcumin 25mg/kg/ day), low (cyclophosphamide 100mg/kg/5 day, curcumin 12.5mg/kg/ day) dosage group.Per 5 days intraperitoneal administrations of cyclophosphamide group once, curcumin group tail every day intravenously administrable once, successive administration 10 days.After the last administration 24 hours, to weigh, the eye socket blood sampling utilizes full-automatic determination of blood cell instrument to measure leukocyte count.Adopt student t test that curcumin emulsion group and injection group tumor are heavily compared.

Table 6. curcumin emulsion is to the Attenuation of chemotherapeutics

Compare with model group: ^*, P＜0.01.Compare with the cyclophosphamide group: ^#, P＜0.05; ^##, P＜0.01

The results are shown in Table 6, after the cyclophosphamide of high dose was administered twice, the mice body weight was starkly lower than model group, and after cyclophosphamide and curcumin merge administration, the mice body weight apparently higher than single to the cyclophosphamide group.Numeration of leukocyte result shows that the high dose cyclophosphamide can cause the murine interleukin number obviously to be reduced, but after the therapeutic alliance of curcumin vein emulsion, leukocyte count is apparently higher than simple cyclophosphamide treatment group.Give curcumin Emulsion merely mice body weight and leukocyte count are not seen obvious influence.These results show that curcumin emulsion can obviously alleviate the toxic and side effects of chemotherapeutic to tumor patient.

Experimental example 4 curcumin emulsions are to the tumor-bearing mice Immune Effects

Prepare curcumin emulsion by preparation example 4 methods.Sterilization back room temperature is placed.Kunming mouse is provided by Shandong Province's natural drug Engineering Technical Research Centre animal center.The strain of H22 Mouse Liver carcinoma is drawn from pharmacological room of Beijing medicine institute.Get the good H22 ascites of all growth conditions that the goes down to posterity mice of going down to posterity, get ascites after, add the long-pending injection normal saline dilution of triploid, be inoculated in 4 Kunming mouse oxters, every 0.2ml.10 mices do not connect tumor and make normal control in addition.Connect after the tumor second day, by body weight mice is divided into 5 groups at random respectively, 10 every group.Be respectively normal group, model group, curcumin emulsion height (50mg/kg/ day), in (25mg/kg/ day), low (12.5mg/kg/ day) group.Curcumin emulsion group tail every day intravenously administrable once, successive administration 10 days.After the last administration 24 hours, the aseptic spleen of getting was placed in the RPMI-1640 of serum-free, is ground to individual cells with piston, crossed 200 order nylon wires, and filtrate 1000rmp * 10min is centrifugal.In the sedimentation cell cell, add 1ml sterilization ultra-pure water splitting erythrocyte, with centrifugal collection splenocyte behind 2 times of D-hanks liquid adjustment of 1ml osmotic pressure.It is inferior to give a baby a bath on the third day after its birth with RPMI-1640.

The NK cytotoxic activity: each experimental group splenocyte is the effector lymphocyte, is adjusted into 2 * 10 with 1640 culture medium that contain 10% calf serum ⁷Cell/ml, every hole adds 100 μ l splenocyte suspensions.As target cell, be adjusted into 2 * 10 with mice YAC-1 lymphoma cell with 1640 culture medium that contain 10% calf serum ⁶Cell/ml, every hole adds target cell suspension 100 μ l, every group establish 3 parallel.The nature releasing tube adds YAC-1 cell suspension 100 μ l, contains 1640 culture medium, the 100 μ l of 10% calf serum.The every pipe of maximum releasing tube adds YAC-1 cell suspension 100 μ l, contains the RPMI-1640 100 μ l of 1%TritonX 100 and 10% calf serum.Above-mentioned each test group is put 37 ℃, 5%CO ₂Incubator in, saturated humidity is cultivated 18h, 1000 rev/mins, centrifugal 5min gets supernatant.Measure the lactic acid dehydrogenase activity of supernatant with lactic acid glue hydrogen enzyme reagent kit (Zhongsheng Beikong Biological Science ﹠ Technology Co., Ltd., Beijing).Calculate the NK cell killing activity.

The splenocyte conversion reaction: above-mentioned splenocyte suspension is adjusted into 1 * 10 with 1640 culture medium that contain 10% calf serum ⁶Cell/ml.Every hole adds spleen born of the same parents suspension 100 μ l in 96 porocyte culture plates, contains the RPMI-1640 of ConA in addition, and making the ConA final concentration is 10 μ g/ml, and other establishes and does not add the ConA group in contrast.Three parallel holes are established in every processing.Culture plate is put 5% CO ₂Behind the incubator 68h, every MTT solution that contains 10mg/ml that also adds.Continue to cultivate 4h.1000rpm * 10min abandons supernatant, adds the bluish violet crystallization that 100 μ lDMSO dissolving generates, and (USA) 570nm reads at the place OD value to microplate reader for Synergy HT, Bio-Tek.The result represents with transformation index.

Serum il-2 level determination: above-mentioned administration tumor-bearing mice, after the last administration 24 hours, eye socket blood sampling 0.5ml, the centrifugal collection serum of 3000rpm * 10min.(Genzyme USA) detects IL-2 level in the serum to measure test kit with IL-2ELISA.Calculate IL-2 content according to standard curve.

Table 7. curcumin emulsion is to the tumor-bearing mice Immune Function

Compare with normal group: ^*, P＜0.01.Compare with model group: ^#, P＜0.05; ^##, P＜0.01.

The result shows that tumor-bearing mice NK cytoactive, lymhocyte transformation rate and IL-2 level all obviously reduce than the normal group mice, and after giving curcumin Emulsion, above-mentioned condition all has clear improvement.Illustrate that curcumin Emulsion can strengthen the tumor patient immunologic function.

Experimental example 5 curcumin emulsion injections are to the potentiation of cyclophosphamide.

Experimental technique: get the mice of going down to posterity of the U14 cervical cancer ascites of inoculating 10 days, get milky ascites with asepsis injector behind the sterilization abdominal part, add the injection normal saline dilution of 3 times of volumes.80 of mices, behind the armpit skin of sterilization right side, the above-mentioned tumor cell suspension 0.2ml of subcutaneous vaccination.Be divided into 8 groups with tumor-bearing mice at random by body weight next day after connecing tumor, 10 every group.Be respectively model group, cyclophosphamide group, curcumin emulsion injection senior middle school low dose group, the high, normal, basic dosage of curcumin emulsion injection and cyclophosphamide combination group.The mice routine is raised to lotus tumor the 6th day, in the time of can touching Subcutaneous tumor, gives relative medicine by dosage shown in the table 8.Cyclophosphamide only once in intraperitoneal administration, the administration of curcumin emulsion injection tail vein injection every day, successive administration 5 days, administration volume are the 25ml/kg body weight.Behind the last administration 24h, take off cervical vertebra and put to death mice, strip tumor tissue and weigh, calculate tumour inhibiting rate.

Utilize two-way analysis of variance to carry out statistical analysis, determine the potentiation that curcumin emulsion injection and cyclophosphamide combined are used.Experimental result:

Table 8 curcumin emulsion injection is to the potentiation of cyclophosphamide

Compare with model group: *, P＜0.05; *, * *, P＜0.01.

Experimental result sees Table 8, and the result shows that when curcumin emulsion injection and cyclophosphamide combined application, antitumor activity obviously improves.Can repeat the two-way analysis of variance result and show, senior middle school's dosage curcumin emulsion injection and cyclophosphamide combined application table reveal obvious synergistic effect (P＜0.05).

Experimental example 6 curcumin emulsion injections press down the potentiation of tumor to cisplatin

Experimental technique: get the mice of going down to posterity of the U14 cervical cancer ascites of inoculating 10 days, get milky ascites with asepsis injector behind the sterilization abdominal part leather belly, add the injection normal saline dilution of 3 times of volumes.80 of mices, behind the armpit skin of sterilization right side, the above-mentioned tumor cell suspension 0.2ml of subcutaneous vaccination.Be divided into 8 groups with tumor-bearing mice at random by body weight next day after connecing tumor, 10 every group.Be respectively model group, cisplatin group (CP), curcumin emulsion injection senior middle school low dose group, the high, normal, basic dosage of curcumin emulsion injection and cisplatin combined set of applications.The mice routine is raised to lotus tumor the 6th day, in the time of can touching Subcutaneous tumor, gives relative medicine by dosage shown in the table 9.Cisplatin intraperitoneal administration every day, 1mg/kg.Curcumin emulsion injection tail every day intravenously administrable, successive administration 5 days, the administration volume is the 25ml/kg body weight.Behind the last administration 24h, take off cervical vertebra and put to death mice, strip tumor tissue and weigh, calculate tumour inhibiting rate.Utilize two-way analysis of variance to carry out statistical analysis, determine the potentiation that curcumin emulsion injection and cyclophosphamide combined are used.Experimental result:

Table 9 curcumin emulsion injection is to the potentiation of cisplatin

Compare with model group: *, P＜0.05; *, P＜0.05;

Experimental result sees Table 9.The result shows, when curcumin emulsion injection and cisplatin combined application, can obviously improve the antitumor activity of cisplatin.Two-way analysis of variance is the result show, senior middle school's dosage curcumin emulsion injection and cyclophosphamide combined application table reveal obvious synergistic effect (P＜0.05).