CN102133185A - Nobiliside A invisible nano liposome intravenous injection and preparation method thereof - Google Patents
Nobiliside A invisible nano liposome intravenous injection and preparation method thereof Download PDFInfo
- Publication number
- CN102133185A CN102133185A CN2011100637054A CN201110063705A CN102133185A CN 102133185 A CN102133185 A CN 102133185A CN 2011100637054 A CN2011100637054 A CN 2011100637054A CN 201110063705 A CN201110063705 A CN 201110063705A CN 102133185 A CN102133185 A CN 102133185A
- Authority
- CN
- China
- Prior art keywords
- nobiliside
- preparation
- stealthy
- intravenous fluid
- polyethylene glycol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to the technical field of medicine, in particular to an invisible nano liposome intravenous injection preparation containing Nobiliside A with antitumor activity and a preparation method thereof. The invisible nano liposome intravenous injection preparation comprises the following components in proportion: 0.01-100mg/mL of Nobiliside A, 1-50mg/mL of phospholipid, 0.01-20mg/mL of cholesterol, 0.1-20mg/mL of PEG (polyethylene glycol) lipid and balance of water for injection. Indicated by animal experiments, compared with the common Nobiliside A liposome intravenous injection preparations, the Nobiliside A invisible nano liposome intravenous injection preparation provided by the invention has a longer half life and small toxic side effects on the liver and spleen, and the tumor inhibition rate is improved by about 30% compared with the common Nobiliside A liposome intravenous injection. The preparation method of the Nobiliside A invisible nano liposome intravenous injection provided by the invention is simple, can effectively improve the tumor inhibition rate of Nobiliside A and reduce the toxic side effects of the Nobiliside A.
Description
Technical field
The present invention relates to medical technical field, is a kind of stealthy nanometer liposome intravenous injection liquid formulation and preparation method thereof with Nobiliside A of anti-tumor activity.
Background technology
Nobiliside A is that a kind of triterpene saponin componds with anti-tumor activity that extracts from Holothuria nobilis Selenka (sees patent of invention " antitumoral compounds Nobiliside A and monoacylated derivant thereof in the Holothuria nobilis Selenka " for details; patent publication No.: CN1740185), chemical structural formula is as follows:
Nobiliside A not only has the good cell toxic action, forms inhibitory action but also have significant new vessels, and the accent of energy induced tumor endotheliocyte and tumor cell is died.Nobiliside A has been made the conventional liposome intravenous fluid and applied for that patent (sees patent of invention " monoacylated composite lipidosome intravenous fluid of antitumoral compounds Nobiliside A and preparation method thereof " for details; patent publication No.: CN1739543); experimental result shows: Nobiliside A conventional liposome intravenous fluid can significantly reduce the hemolytic of Nobiliside A inside and outside, has improved stimulation and the proinflammatory effect of Nobiliside A to blood vessel.Yet, after the Nobiliside conventional liposome enters in the body, easily by reticuloendothelial system (RES) picked-up, removing, the half-life of Nobiliside conventional liposome in blood lacked (half-life is generally about 10h), also easily the abundant organ (as liver and spleen) of RES is caused cumulative toxicity, the targeting of target tissue is poor, and tumor killing effect is undesirable.
Stealthy nanometer liposome is called long-circulating nanoliposome again, and this lipoid plastid particle diameter is little, usually about 100nm.Surface of liposome can form sterically hindered, the opsonic action that effectively stops plasma protein, make liposome can escape the discovery of RES and catch, the blood circulation time of prolong drug helps entering tumor tissues, and promote its infiltration and delay effect (EPR effect) to tumor tissues, messenger drug object height concentration, be accumulated in the tumor tissues for a long time, improve therapeutic effect (Tao Tao, the clinical progress of liposome delivery system [J]. pharmaceutical services and research, 2008,8 (2): 84-88).Yet do not see the relevant report of Nobiliside A being made stealthy nanometer liposome intravenous fluid so far.
Summary of the invention
The invention provides that long half time in a kind of body, toxic and side effects are little, good effect, the stealthy nanometer liposome intravenous injection of the Nobiliside A liquid formulation that tumor had medicine passive target function.
The component and the proportioning of the stealthy nanometer liposome intravenous fluid of Nobiliside A of the present invention are as follows:
Preferably:
Wherein, phospholipid is selected from one or more in soybean lecithin, Ovum Gallus domesticus Flavus lecithin, distearoyl phosphatidylcholine, dipalmitoyl phosphatidyl choline, the dimyristoyl phosphatidyl choline, preferably soya lecithin and/or Ovum Gallus domesticus Flavus lecithin.
The PEG lipid is selected from one or more in Polyethylene Glycol-DSPE, polyethylene glycol-caprolactone, polyethylene glycol-Acetic acid, hydroxy-, bimol. cyclic ester lactide, polyethylene glycol-lactic acid, polyethylene glycol-cetyl cyanoacrylate, polyoxyethylene fatty acid ether, the polyoxyethylene methyl Oleum Ricini ether, preferred Polyethylene Glycol-DSPE.
Preparation method is:
(1) preparation class lipoprotein solution:
By proportioning phospholipid, cholesterol and PEG lipid are dissolved in the 50mL chloroform, the volume ratio by chloroform and dehydrated alcohol is 1 then: 2-1: 4 add dehydrated alcohol, get the class lipoprotein solution;
(2) preparation class membrane of lipoprotein:
The class lipoprotein solution is placed on the rotary evaporator,, removes organic facies chloroform and dehydrated alcohol at 50 ℃ of rotation reduction vaporization 1-1.5h, the class lipoprotein solution one deck homogeneous film;
(3) preparation Nobiliside A hidden liposome suspension:
Water for injection by proportioning adding Nobiliside A aqueous solution and surplus makes the calm wall of class membrane of lipoprotein elute and be made into suspension, promptly gets Nobiliside A hidden liposome suspension;
(4) the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
With Nobiliside A hidden liposome suspension high pressure homogenize routinely, the filtering with microporous membrane sterilization promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Key of the present invention is to adopt the PEG lipid to prepare the stealthy nanometer liposome intravenous fluid of Nobiliside A.The PEG lipid contains hydrophilic group, the lipid-modified liposome of PEG forms sterically hinderedly at surface of liposome, make that stealthy nanometer liposome is difficult for being discerned and absorbing by RES, therefore the circulation time of stealthy nanometer liposome in blood prolongs, and makes medicine gather tumor tissues more; The lipid-modified liposome of PEG makes stealthy nanometer liposome particle diameter less than 100nm, also increases the stability of the stealthy nanometer liposome intravenous fluid of Nobiliside A.
Zoopery shows that the stealthy nanometer liposome intravenous injection of Nobiliside A of the present invention liquid formulation blood halflife reaches 15.93 hours; Toxic and side effects to liver and spleen is little, has reduced 3 times and 4 times respectively at the Nobiliside of liver and spleen A concentration ratio Nobiliside A conventional liposome intravenous injection liquid formulation; Tumour inhibiting rate has improved about 30% than Nobiliside A conventional liposome intravenous fluid.
The stealthy nanometer liposome intravenous injection of Nobiliside A of the present invention liquid and preparation method thereof is simple, can effectively improve the tumour inhibiting rate of Nobiliside A, reduces the toxic and side effects of Nobiliside A.
Description of drawings
Fig. 1 is that the stealthy nanometer liposome intravenous fluid of Nobiliside A is at the intravital blood drug level-time plot of rat.
Fig. 2 is the drug level-time diagram of the stealthy nanometer liposome intravenous fluid of Nobiliside A in mouse liver.
Fig. 3 is the drug level-time diagram of the stealthy nanometer liposome intravenous fluid of Nobiliside A in mouse spleen.
Fig. 4 is the curve chart of the stealthy nanometer liposome intravenous fluid of Nobiliside A to the tumor-bearing mice tumour inhibiting rate.
The specific embodiment
Embodiment 1: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
Soybean lecithin 0.5g, cholesterol 0.01g and Polyethylene Glycol-DSPE 0.5g, polyethylene glycol-caprolactone 0.2g, polyethylene glycol-Acetic acid, hydroxy-, bimol. cyclic ester lactide 0.3g are dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 2 adding dehydrated alcohol 100mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 10mL contains 0.01gNobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 100mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Embodiment 2: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
Ovum Gallus domesticus Flavus lecithin 0.4g, cholesterol 0.002g and Polyethylene Glycol-DSPE 0.015g, polyethylene glycol-lactic acid 0.005g are dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 3 adding dehydrated alcohol 150mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 10mL contains 0.02g Nobiliside A elutes thin film and make suspension, promptly gets the hidden liposome suspension of Nobiliside A; The aqueous solution for injection that adds surplus is settled to 200mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Embodiment 3: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
Soybean lecithin 1.0g, Ovum Gallus domesticus Flavus lecithin 0.5g, cholesterol 0.8g and Polyethylene Glycol-DSPE 0.3g, polyethylene glycol-cetyl cyanoacrylate 0.1g, polyoxyethylene fatty acid ether 0.1g are dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 4 adding dehydrated alcohol 200mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 20mL contains 1.0g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 100mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Embodiment 4: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
Soybean lecithin 1.0g, stearoyl phosphatidylcholine 0.5g, dipalmitoyl phosphatidyl choline 0.5g, cholesterol 1.0g and Polyethylene Glycol-DSPE 1.0g are dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 3 adding dehydrated alcohol 150mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.5h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 10mL contains 5.0g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 50mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Embodiment 5: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
Soybean lecithin 0.9g and dimyristoyl phosphatidyl choline 0.1g, cholesterol 0.5g and Polyethylene Glycol-DSPE 0.3g, methyl Oleum Ricini ether 0.1g are dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 3 adding dehydrated alcohol 150mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 10mL contains 0.05g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 100mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Embodiment 6: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
Ovum Gallus domesticus Flavus lecithin 1.7g, distearoyl phosphatidylcholine 0.8g, cholesterol 0.75g and Polyethylene Glycol-DSPE 0.45g, polyethylene glycol-caprolactone 0.55g are dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 4 adding dehydrated alcohol 200mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.5h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 20mL contains 2.5g NobilisideA elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 50mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Embodiment 7: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
Soybean lecithin 0.6g, Ovum Gallus domesticus Flavus lecithin 0.2g, distearoyl phosphatidylcholine 0.1g, cholesterol 0.4g and Polyethylene Glycol-DSPE 0.4g are dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 2 adding dehydrated alcohol 100mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 5mL contains 0.04g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 100mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Embodiment 8: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
Soybean lecithin 4.0g, Ovum Gallus domesticus Flavus lecithin 3.5g, cholesterol 3.0g and Polyethylene Glycol-DSPE 2.25g are dissolved in the 50mL chloroform, and the volume ratio by chloroform and dehydrated alcohol is 1: 3 adding dehydrated alcohol 150mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.5h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 15mL contains 3.0g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 150mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get NobilisideA stealth nanometer liposome intravenous fluid.
Embodiment 9: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
Soybean lecithin 0.5g, cholesterol 0.25g and Polyethylene Glycol-DSPE 0.1g, polyethylene glycol-caprolactone 0.1g are dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 3 adding dehydrated alcohol 150mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 5mL contains 0.02g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 100mL, and high pressure homogenize routinely promptly get Nobiliside A stealth nanometer liposome intravenous fluid then.
Embodiment 10: the stealthy nanometer liposome parenteral solutions of preparation Nobiliside A
With soybean lecithin 2.0g, cholesterol 1.0g and Polyethylene Glycol-DSPE 2.0g, be dissolved in the 50mL chloroform, be to add dehydrated alcohol 150mL at 1: 3 by the volume ratio of chloroform and dehydrated alcohol again, the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.2h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 10mL contains 1.0g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of NobilisideA; The water for injection that adds surplus is settled to 200mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Embodiment 11: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
Soybean lecithin 1.0g, cholesterol 0.5g and polyethylene glycol-caprolactone 0.2g, Polyethylene Glycol poly (glycolide-co-lactide) 0.1g are dissolved in the 50mL chloroform, and the volume ratio by chloroform and dehydrated alcohol is 1: 2 adding dehydrated alcohol 100mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.5h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 5mL contains 0.03g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 100mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get NobilisideA stealth nanometer liposome intravenous fluid.
Embodiment 12: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
Soybean lecithin 3.5g and dimyristoyl phosphatidyl choline 0.5g, cholesterol 2.0g and Polyethylene Glycol-DSPE 0.75g, methyl Oleum Ricini ether 0.25g are dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 4 adding dehydrated alcohol 200mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.4h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 5mL contains 0.2g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 200mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Embodiment 13: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
Soybean lecithin 0.6g and dimyristoyl phosphatidyl choline 0.6g, cholesterol 0.55g and Polyethylene Glycol-DSPE 0.3g, methyl Oleum Ricini ether 0.1g are dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 3 adding dehydrated alcohol 150mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.3h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 10mL contains 0.05g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 100mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Embodiment 14: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
Soybean lecithin 1.25g, Ovum Gallus domesticus Flavus lecithin 1.0g, cholesterol 0.15g and Polyethylene Glycol-DSPE 0.6g, polyethylene glycol-caprolactone 0.3g are dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 4 adding dehydrated alcohol 200mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.2h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 10mL contains 0.15g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 150mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Embodiment 15: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
With Ovum Gallus domesticus Flavus lecithin 0.8g, distearoyl phosphatidylcholine 0.7g, cholesterol 0.55g and Polyethylene Glycol-DSPE 0.3g, polyoxyethylene fatty acid ether 0.2g is dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 4 adding dehydrated alcohol 200mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.5h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 5mL contains 0.055g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 100mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Embodiment 16: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
With Ovum Gallus domesticus Flavus lecithin 0.1g, distearoyl phosphatidylcholine 0.1g, cholesterol 0.004g and polyethylene glycol-lactic acid 0.0035g, polyoxyethylene methyl Oleum Ricini ether 0.0005g is dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 4 adding dehydrated alcohol 200mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.5h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 5mL contains 0.004g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 200mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Execute example 17: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
With distearoyl phosphatidylcholine 0.8g, dipalmitoyl phosphatidyl choline 0.8g, cholesterol 0.8g and polyethylene glycol-lactic acid 0.35g, polyethylene glycol-cetyl cyanoacrylate 0.25g is dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 3 adding dehydrated alcohol 150mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.5h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 10mL contains 0.06g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 100mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Execute example 18: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
With soybean lecithin 1.1g, Ovum Gallus domesticus Flavus lecithin 0.5g, distearoyl phosphatidylcholine 0.1g, dipalmitoyl phosphatidyl choline 0.1g, cholesterol 0.6g and polyethylene glycol-lactic acid 0.3g, polyethylene glycol-caprolactone 0.6g is dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 3 adding dehydrated alcohol 150mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.5h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 10mL contains 0.12g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 150mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Execute example 19: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
With soybean lecithin 0.8g, Ovum Gallus domesticus Flavus lecithin 0.5g, distearoyl phosphatidylcholine 0.4g, cholesterol 0.7g and polyethylene glycol-lactic acid 0.3g, polyethylene glycol-caprolactone 0.2g is dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 3 adding dehydrated alcohol 150mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.5h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 10mL contains 0.08g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 100mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Execute example 20: the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
With soybean lecithin 0.5g, Ovum Gallus domesticus Flavus lecithin 0.5g, distearoyl phosphatidylcholine 0.5g, dimyristoyl phosphatidyl choline 0.5g, cholesterol 1.2g and Polyethylene Glycol-DSPE 0.8g, polyethylene glycol-caprolactone 0.8g is dissolved in the 50mL chloroform, volume ratio by chloroform and dehydrated alcohol is 1: 4 adding dehydrated alcohol 200mL again, gets the class lipoprotein solution; The class lipoprotein solution is placed on the rotary evaporator, remove organic facies at 50 ℃ of rotation reduction vaporization 1.4h, the class lipoprotein solution one deck homogeneous film, the aqueous solution that adding 10mL contains 0.1g Nobiliside A elutes thin film and make suspension, is the hidden liposome suspension of Nobiliside A; The water for injection that adds surplus is settled to 100mL, high pressure homogenize routinely then, and the sterilization of 0.22 μ m filtering with microporous membrane promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Zoopery
1.Nobiliside the stealthy nanometer liposome intravenous fluid of A is tested in pharmacokinetics in rats
Material and reagent: the stealthy nanometer liposome intravenous fluid of Nobiliside A (by embodiment 1 preparation, down together); Rosuvastain calcium (Shanghai Institute of Pharmaceutical Industry); Heparin sodium (Shanghai No.1 Bio-Chemical Pharmacetical Industry Co., Ltd); Methanol (chromatographically pure, Sigma-Aldrich, USA).
Mass spectrum condition: API 3000 type triple quadrupole tandem mass spectrometers, electric spray ion source (Turbo Ionspray) and Analyst 1.4 data handling systems (u.s.a. applied biosystem company limited); Agilent 1100 liquid chromatographic systems (U.S. Agilent company); Ionspray voltage is 3.50kV; 400 ℃ of atomization temperatures; Atomization gas (N
2) be 900Lh
-1Auxiliary gas (the N of heating
2) be 8.0Lmin
-1Curtain gas CUR is 8.0Lmin
-1Collision gas (N
2) 4.0Lmin
-1Be 0.2s sweep time; Adopt the anion mode to detect; Scan mode is multiple-reaction monitoring (MRM); The ionic reaction that is used for quantitative analysis is respectively m/z1183.4-m/z 96.9amu (Nobiliside A) and m/z 421.2＞m/z 101.0amu (interior mark: rosuvastain calcium).
Chromatographic condition: chromatographic column: Waters Xbridge C
18(2.1 * 50mm, 3 μ m); Pre-column: Waters Xbridge C
18(2.1 * 10mm, 3 μ m); Mobile phase: methanol: water (70: 30, V/V); Flow velocity: 0.2mL/min; Column temperature: 30 ℃.
Animal: the SD rat, male, 200 ± 20g is provided by Shanghai Slac Experimental Animal Co., Ltd., and production licence number SCXK (Shanghai) is (2007-2005).
Experimental technique: get 10 of SD male rats, be divided into two groups at random, 5 every group, be respectively the stealthy nanometer liposome intravenous fluid of Nobiliside A conventional liposome intravenous fluid group and Nobiliside A group, dosage is by body weight tail vein injection 1mg/kg, respectively at 5min after the administration, 10min, 15min, 30min, 45min, 1h, 2h, 4h, 8h, 12h, 24h, 48h and 72h eye socket are got blood 0.3mL, place in the centrifuge tube that is added with heparin sodium, shake up, the centrifugal 5min of 15000rpm, get 100 μ l plasma samples and put in the centrifuge tube, add methanol 20 μ l, add inner mark solution 500 μ l (200ng/mL), vortex 1min, the centrifugal 5min of 15000rpm gets supernatant and carries out the LC/MS/MS analysis, measures the content of Nobiliside A respectively, average and map, the results are shown in Figure 1.
As seen from Figure 1, the area under the drug-time curve (AUC) of the stealthy nanometer liposome intravenous fluid of Nobiliside A is 8 times of AUC of Nobiliside A conventional liposome intravenous fluid, and Nobiliside A blood is eliminated the half-life and extended to 22.93h by the 10.58h of Nobiliside A conventional liposome intravenous fluid.Explanation can make the blood circulation time of Nobiliside A prolong in Nobiliside A preparation cost invention preparation.
2.Nobiliside the stealthy nanometer liposome intravenous fluid of A is in the intravital tissue distribution experiment of mice
Material and reagent, mass spectrum condition, chromatographic condition are the same.
Animal: the ICR mice, male, body weight 20 ± 2g is provided by Shanghai Slac Experimental Animal Co., Ltd., and production licence number SCXK (Shanghai) is (2007-2005).
Get 50 of ICR mices, be divided into two groups at random, every group 25, be respectively the stealthy nanometer liposome intravenous fluid of Nobiliside A conventional liposome intravenous fluid group and Nobiliside A group, dosage is by body weight tail vein injection 1mg/kg, respectively at 5min after the administration, 1h, 8h, 24h and 48h respectively put to death 5, get every Mouse Liver respectively, each 100mg of spleen, respectively organize shred respectively after, each adds the homogenate routinely of 0.3mL normal saline, gets 100 μ l homogenates and places centrifuge tube, add methanol 20 μ l, mark rosuvastain calcium methanol solution (200ng/mL) 500 μ l in adding, vortex 1min, the centrifugal 5min of 15000rpm, get supernatant and carry out the LC/MS/MS analysis, measure the content of NobilisideA respectively, get the meansigma methods and the mapping of each time point, the results are shown in Figure 2, Fig. 3.
By Fig. 2 and Fig. 3 as seen, in liver that is rich in reticuloendothelial cell and spleen, the mice Nobiliside A content of the stealthy nanometer liposome intravenous fluid of injection Nobiliside A group is starkly lower than Nobiliside A conventional liposome parenteral solutions group, illustrates that preparation of the present invention can reduce the toxic and side effects to liver and spleen.
3.Nobiliside the stealthy nanometer liposome intravenous fluid of A is to the treatment experiment of mice S180 lymphosarcoma
Positive control drug is Cyclophosphamide for injection (CTX), SHANXI POWERDONE PHARMACEUTICAL.,LTD, and product batch number: 20100308,200mg/10mL props up, and dilutes with normal saline during use.
Animal: the ICR mice, male, body weight 20 ± 2g is provided by Shanghai Slac Experimental Animal Co., Ltd., and production licence number SCXK (Shanghai) is (2007-2005).
Tumor strain: mice S180 lymphosarcoma (being provided) by Chinese Academy of Sciences's Shanghai medicine.
With the mouse lymph lymphoma S180 tumor strain abdominal cavity inoculation of going down to posterity routinely, inoculate about 1 week the dislocation of mice cervical vertebra is put to death, extract ascites out, with the normal saline dilution, make 1-2 * 10
7Individual/mL cell suspension; Get 80 of ICR mices, give every mice right fore axillary region subcutaneous injection tumor strain diluent 0.2mL respectively, treat that the tumor body is long to 150mm
3About, be divided into eight groups at random, every group 10, be respectively negative control group, positive controls, Nobiliside A conventional liposome parenteral solutions is low, in, the stealthy nanometer liposome parenteral solutions of high dose group and Nobiliside A is low, in, high dose group, the height of stealthy nanometer liposome intravenous fluid of Nobiliside A and Nobiliside A conventional liposome parenteral solutions, in, the low dose group dosage all is followed successively by by body weight 2mg/kg, 1mg/kg, 0.5mg/kg, the positive controls dosage is CTX 20mg/kg, the tail vein injection administration, once a day, each 0.2mL, negative control group tail vein injection equal-volume normal saline, successive administration 7 days is observed tumor bulk-growth situation.After the drug withdrawal 24 hours, put to death animal, weigh respectively, tumor is heavy, calculates the suppression ratio of medicine to tumor according to following formula:
Tumour inhibiting rate %=(the average tumor of the average tumor weight/blank group of 1-administration group is heavy) * 100%
The therapeutic evaluation standard: tumor control rate＜40% is invalid; Tumor control rate 〉=40%, and processing p＜0.05 is effective by statistics.With the tumor control rate mapping, see Fig. 4.Among the figure, CTX represents positive controls; C low, C middle, C high represent successively Nobiliside A conventional liposome intravenous fluid low, in and high dose group; S low, S middle, S high represent successively the stealthy nanometer liposome intravenous fluid of Nobiliside A low, in and high dose group.
As seen from Figure 4, the tumour inhibiting rate of the basic, normal, high dosage group of the stealthy nanometer liposome parenteral solutions of Nobiliside A is followed successively by 38.9%, 45.1% and 63.7%, be significantly higher than the tumour inhibiting rate 10.7% of the basic, normal, high dosage group of Nobiliside A conventional liposome parenteral solutions, 21.2% and 32.3, illustrate that the stealthy nanometer liposome parenteral solutions of Nobiliside A of the present invention can significantly improve the suppression ratio of Nobiliside A to tumor.
Claims (4)
1. the stealthy nanometer liposome intravenous fluid of a Nobiliside A, component and proportioning are as follows:
Wherein, phospholipid is selected from one or more in soybean lecithin, Ovum Gallus domesticus Flavus lecithin, distearoyl phosphatidylcholine, dipalmitoyl phosphatidyl choline, the dimyristoyl phosphatidyl choline;
The PEG lipid is selected from one or more in Polyethylene Glycol-DSPE, polyethylene glycol-caprolactone, polyethylene glycol-Acetic acid, hydroxy-, bimol. cyclic ester lactide, polyethylene glycol-lactic acid, polyethylene glycol-cetyl cyanoacrylate, polyoxyethylene fatty acid ether, the polyoxyethylene methyl Oleum Ricini ether.
2. by the stealthy nanometer liposome intravenous fluid of the described Nobiliside A of claim 1, it is characterized in that said phospholipid is soybean lecithin and/or Ovum Gallus domesticus Flavus lecithin, said PEG lipid is Polyethylene Glycol-DSPE.
3. by claim 1 or the stealthy nanometer liposome intravenous fluid of 2 described Nobiliside A, it is characterized in that component and proportioning are as follows:
4. the preparation method of claim 1 or the stealthy nanometer liposome intravenous fluids of 2 or 3 described Nobiliside A, step is as follows:
(1) preparation class lipoprotein solution:
By proportioning phospholipid, cholesterol and PEG lipid are dissolved in the 50mL chloroform, the volume ratio by chloroform and dehydrated alcohol is 1 then: 2-1: 4 add dehydrated alcohol, get the class lipoprotein solution;
(2) preparation class membrane of lipoprotein:
The class lipoprotein solution is placed on the rotary evaporator,, removes organic facies chloroform and dehydrated alcohol at 50 ℃ of rotation reduction vaporization 1-1.5h, the class lipoprotein solution the class membrane of lipoprotein;
(3) preparation Nobiliside A hidden liposome suspension:
Water for injection by proportioning adding Nobiliside A aqueous solution and surplus makes the calm wall of class membrane of lipoprotein elute and be made into suspension, promptly gets Nobiliside A hidden liposome suspension;
(4) the stealthy nanometer liposome intravenous fluid of preparation Nobiliside A
With Nobiliside A hidden liposome suspension high pressure homogenize routinely, the filtering with microporous membrane sterilization promptly get Nobiliside A stealth nanometer liposome intravenous fluid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100637054A CN102133185A (en) | 2011-03-16 | 2011-03-16 | Nobiliside A invisible nano liposome intravenous injection and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100637054A CN102133185A (en) | 2011-03-16 | 2011-03-16 | Nobiliside A invisible nano liposome intravenous injection and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102133185A true CN102133185A (en) | 2011-07-27 |
Family
ID=44293199
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011100637054A Pending CN102133185A (en) | 2011-03-16 | 2011-03-16 | Nobiliside A invisible nano liposome intravenous injection and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102133185A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109758425A (en) * | 2019-03-18 | 2019-05-17 | 中国海洋大学 | A kind of selenka nano liposomes and preparation method thereof for drug administration by injection |
-
2011
- 2011-03-16 CN CN2011100637054A patent/CN102133185A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 《药学与临床研究》 20110228 顾芃等 《PEG化隐形纳米脂质体的研究进展》 第47-50页 1-4 第19卷, 第1期 * |
| 《药学学报》 20081231 熊阳等 《海参皂苷nobiliside A脂质体及其溶血行为的初步研究》 第214－220页 1-4 第43卷, 第2期 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109758425A (en) * | 2019-03-18 | 2019-05-17 | 中国海洋大学 | A kind of selenka nano liposomes and preparation method thereof for drug administration by injection |
| CN109758425B (en) * | 2019-03-18 | 2021-07-23 | 中国海洋大学 | Sea cucumber saponin nano liposome for injection administration and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Pleguezuelos-Villa et al. | A novel ultradeformable liposomes of Naringin for anti-inflammatory therapy | |
| Sun et al. | Synergistic amplification of oxidative Stress–Mediated antitumor activity via liposomal dichloroacetic acid and MOF‐Fe2+ | |
| EP2938332B1 (en) | Sustained-release lipid pre-concentrate of gnrh analogues and pharmaceutical composition comprising the same | |
| Wang et al. | Improved solubility and pharmacokinetics of PEGylated liposomal honokiol and human plasma protein binding ability of honokiol | |
| US8956659B2 (en) | Fulvestrant nanosphere/microsphere and preparative method and use thereof | |
| EP3239132B1 (en) | Cationic lipid | |
| Tong et al. | Enhanced antitumor efficacy and decreased toxicity by self-associated docetaxel in phospholipid-based micelles | |
| Lohumi | A novel drug delivery system: niosomes review | |
| WO2010009186A1 (en) | Liposome formulation having hydrophilic and hydrophobic pharmaceutical compounds co-encapsulated therein | |
| Cao et al. | A combination of a microemulsion and a phospholipid complex for topical delivery of oxymatrine | |
| Tarekegn et al. | Niosomes in targeted drug delivery: some recent advances | |
| CN103923277A (en) | Amphipathic polymer, preparation method thereof, composite nano medicine carrier and preparation method thereof | |
| Chuealee et al. | Bioactivity and toxicity studies of amphotericin B incorporated in liquid crystals | |
| Xu et al. | Preparation of a paclitaxel-loaded cationic nanoemulsome and its biodistribution via direct intratumoral injection | |
| Zheng et al. | Self-delivery nanomedicine to overcome drug resistance for synergistic chemotherapy | |
| Yu et al. | Novel long-circulating liposomes consisting of PEG modified β-sitosterol for gambogic acid delivery | |
| CN107148410B (en) | Cationic lipids for nucleic acid delivery | |
| US20050142182A1 (en) | Long circulating liposome | |
| CN102133185A (en) | Nobiliside A invisible nano liposome intravenous injection and preparation method thereof | |
| Jain et al. | Niosome a novel approach for drug delivery system: an overview | |
| CN1931157A (en) | Polyene taxol liposome and its prepn process | |
| Sun et al. | Enhanced pharmacokinetics and anti-tumor efficacy of PEGylated liposomal rhaponticin and plasma protein binding ability of rhaponticin | |
| KR20220159986A (en) | Ionic Liquids, Solvents, Formulations, and Transdermal Absorbers | |
| CA2933001C (en) | Siloxane-containing phospholipids, compositions and uses thereof | |
| Ramasamy et al. | Nanocochleate—a new drug delivery system |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20110727 |