CN108524471B - Orlistat nanosphere and its preparing the purposes in obesity treating medicine - Google Patents
Orlistat nanosphere and its preparing the purposes in obesity treating medicine Download PDFInfo
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Abstract
The present invention relates to pharmaceutical technology field, in particular to orlistat nanosphere and its purposes in obesity treating medicine is being prepared.The nanosphere includes orlistat and selected from vitamin A methacrylate -2- methylacryoyloxyethyl phosphocholine copolymer or vitamin E methacrylate -2- methylacryoyloxyethyl phosphocholine copolymer or vitamin D2One of methacrylate -2- methylacryoyloxyethyl phosphocholine copolymer copolymer.The encapsulation rate of the nanosphere is 82.67~93.83%.Animal experiment is shown, after nanosphere orally administration rat the absorption level of each vitamin be apparently higher than orlistat and vitamin stagger the time (2 hours) oral administration vitamin absorption it is horizontal, and it is on close level with the absorption of the vitamin after orally administration vitamin independent under fasting, the missing of liposoluble vitamin caused by oral orlistat can be effectively made up, realizes safe fat-reducing purpose.
Description
Technical field
The present invention relates to pharmaceutical technology field, in particular to a kind of orlistat nanosphere and its fat in preparation treatment
Purposes in disease drug.
Background technique
Orlistat (orlistat) is to research and develop lipase inhibitor class slimming drugs, trade name by company, Roche Group
Xenical, the 1990s end take the lead in listing in America and Europe, are eaten in Discussion on Chinese Listed, and in 2005 by China within 2001
The approval of product Drug Administration switchs to non-prescribed medicine.Entitled N- formyl-L-Leu (the s) -1- of its chemistry [(2s, 3s) -3- oneself
- 4 oxygroup -2- glycidyl methyl of base] ten diester, also referred to as tetrahydrolipostatin (Tetrahydrolipstatin, THL) they are a kind of
Semi-synthetic lipstatin derivative, chemical structural formula are as shown below:
The serine residue of orlistat and stomach pancreatic lipase forms covalence key, can not will be in food so that enzyme be made to inactivate
Triglyceride hydrolysis at absorbable fatty acid, and then reduce the uptake of fat.Currently, orlistat is unique both at home and abroad
The chemical slimming drugs for not influencing appetite, not acting on central nervous system, security features are superior, and Bray GA is in " lancet "
Orlistat is described as " most safe " in the article for the entitled Management of Obesity that magazine is delivered
(safest) slimming drugs.However, the oral preparation of orlistat is for there is also have many deficiencies when treating obesity
Place, one of them is exactly the loss of liposoluble vitamin.Due to orlistat can reduce and meanwhile take include vitamin A, D with
The absorption of liposoluble vitamin including E, therefore must or after orlistat was with meal medication 2 hours or slept during its medication
The preceding suitable liposoluble vitamin of supplement.But under modern society quickly rhythm of life, many patients can forget supplement dimension life
Element and cause a series of adverse reactions.
The definition of nanotechnology (Nanotechnology) is " production or processing nanometer materials or manipulation nanoscopic objects
Ability ", it is just soon of interest by pharmacy scientific and technical personnel since its rise, and be applied to drug delivery (Drug
Delivery) field, wherein the administration nano-drug administration system that can be used for oral drugs includes: liposome, polymer nanocomposite ball/nanometer
Capsule, solid lipid nano granule, micro emulsion, nano-emulsion, self-emulsifying nanometer cream, polymer micelle, dendritic copolymer, nano medicine crystal
Deng.Oral administration nano-drug administration system has the following characteristics that the partial size of 1. drug bearing microsphere within the scope of 10~1000nm, and specific surface area is aobvious
Increase；2. increasing drug solubility；3. improving the dissolution rate of drug；4. the gastrointestinal tract mucosa that enhancing carries powder is sticked
Property；5. enhancing the gastrointestinal tract stability of drug；6. enhancing carries powder in the residence time and face of action position or absorption site
Product；7. enhance drug crosses over Mucosa Barrier ability；8. improving the oral administration biaavailability of drug；9. certain oral administration nanometer administrations
There are also slow-release controlled-release, target functions etc. for system.
Andrej Dolenc et al. (International Journal of Pharmaceutics, 2010, (396) 1
~2:149~155) by orlistat disperse Tween-80, PVP K-30, poloxamer-188 and lauryl sodium sulfate
In the aqueous solution of various stabilizers, and the orlistat for being prepared for Nano grade by melting emulsion process, the Nano grade
The In Vitro Dissolution rate of orlistat is significant to be higher than bulk pharmaceutical chemicals.
Chinese patent CN107412196A discloses a kind of nanometer being made of orlistat and polyethylene glycol-polycaprolactone
Microballoon, the microballoon change the hydrophobic property of orlistat, and by the sustained releasing character of microballoon, can be used for preparing antineoplastic
Object.
Temporary incapability, which provides orlistat simultaneously and had in single pharmaceutical preparation, in the prior art fully absorbs level
Liposoluble vitamin nanometer product technical teaching.
Summary of the invention
Liposoluble vitamin caused by oral orlistat is effectively compensated for the purpose of the present invention is to provide one kind to lack
It loses, realizes the orlistat nanosphere of safe fat-reducing purpose.
Another object of the present invention is that it is overweight and fat in preparation treatment to provide a kind of orlistat nanosphere
Purposes in disease drug.
In order to realize the above object, firstly, present invention reaction route according to shown in the following figure is synthetically prepared and is total to
Polymers 1a~1c, and structural identification has been carried out to copolymer 1 a~1c.
In upper reaction equation, compound 2 is methacrylic acid, and CAS 79-41-4 is one of common industrial chemicals.
Compound 4 is 2- methylacryoyloxyethyl phosphocholine (MPC), can be commercial product, see also
Ishihara K et al.(Polym J 1990；22:355-360.) and Ueda T et al. (Polym J 1992；24:
1259-1269.) method disclosed by is prepared.
ROH is selected from VitAVitE or vitamin D2One of liposoluble vitamin.
In upper reaction equation, compound 2 and ROH carry out acylation reaction, generate compound 3a~3c respectively.
Reaction reagent, condition and the operating method of the step a are as follows: N, N '-dicyclohexyl diimine (DCC) or 4- diformazan
Aminopyridine (DMAP), petroleum ether, 50~60 DEG C.The step is conventional acylation reaction, operation can according to but be not limited to hear
" the one-step method that " drug synthetic reaction " (publication Chemical Industry Press's in April, 2017) of tough chief editor and Liu Xianghong et al. are delivered
Synthesize Retinol Palmitate " (Chemical Industry in Guangzhou, the 14th phase of volume 39 in 2011,63~64) disclose method carry out.Described
Methacrylic acid: the molar ratio of ROH is between 1:1~2:1.
Further, compound 3a~3c is reacted with compound 4 generates copolymer 1 a~1c.
Reaction reagent, condition and the operating method of the step b are as follows: azodiisobutyronitrile (AIBN), methanol/tetrahydrofuran
(MeOH/THF).The Preparation of that the step operation can be delivered according to Kazuhiko Ishihara et al.
Phospholipid Polymers and Their Properties as Polymer Hydrogel Membranes.
Method disclosed by (Polymer Journal, Volume 22, Issue 5,355~360) carries out, and is disclosed according to the document
Method structural identification is carried out to copolymer 1 a~1c for being prepared.
Specifically, in above-mentioned steps b compound 3a~3c and compound 4 feed ratio confirmation method are as follows: using 1:1 as change
Close the minimum molar feed ratio of each compound and compound 4 in object 3a~3c, and the dissolution with gained copolymer in water
Degree >=50mg/mL is index, it is determined that the maximum molar feed ratio of each compound and compound 4 in compound 3a~3c,
The results are shown in Table 1.
The maximum molar feed ratio of two kinds of reactants in 1 step b of table
| Reactant | Maximum molar feed ratio |
| Compound 3a: compound 4 | 4:1 |
| Compound 3b: compound 4 | 7:3 |
| Compound 3c: compound 4 | 6:4 |
Specifically, the definition of substituent R and corresponding preparation are former in copolymer 1 a~1c structural formula in above-mentioned reaction equation
Material, intermediate product are as shown in table 2 below.
The definition of substituent R and corresponding raw material, intermediate product are prepared in 2 copolymer 1 a~1c structural formula of table
Further, the present invention confirms the structure of copolymer 1 a~1c, specially as follows:
(1)1H-NMR
The present invention uses1H-NMR (400Hz, CDCl3), it is true that structure has been carried out to raw material and the product of aforementioned every single step reaction
Card, and by comparing product and raw material1The similarities and differences of H-NMR spectrum data, to conclude the generation of reaction.
(2) in copolymer 1 a~1c structural formula m:n ratio calculating
The present invention is by calculating copolymer 1 a~1c's1- N in H-NMR map+(CH3)3The peak area of the characteristic peak of hydrogen accounts for
The ratio of total hydrogen peak area, calculates the m:n ratio in copolymer 1 a~1c structural formula.
(3) molecular weight determination of copolymer 1 a~1c
The present invention uses gel permeation chromatography well-known to those skilled in the art (GPC), and using THF as solvent,
Using polystyrene as control, the molecular weight of copolymer 1 a~1c of the various m:n ratios of aforementioned gained is determined.
Further, the structural formula of the copolymer 1 a~1c is as shown below:
Wherein, if the copolymer is copolymer 1 a, the m:n ratio in structural formula is excellent between 62:38~82:18
It is selected as 82:18；
If the copolymer is copolymer 1 b, the m:n ratio in structural formula is between 62:38~76:24, preferably
76:24；
If the copolymer is copolymer 1 c, the m:n ratio in structural formula is between 64:36~68:32, preferably
68:32。
Further, whether there is humidification in order to verify copolymer 1 a~1c to orlistat water solubility, the present invention
People has carried out solubility test of the orlistat in copolymer 1 a~1c aqueous solution, specific to use " Chinese Pharmacopoeia " (2015 editions)
Specified in method determine solubility of the orlistat in the 50mg/mL aqueous solution of aforementioned copolymer 1a~1c, particularly,
The present invention determines orlistat in the maximum 50mg/mL aqueous solution with gained copolymer 1 a~1c under minimum molar feed ratio
Solubility, to demonstrate copolymer 1 a~1c to the water-soluble humidification of orlistat.
Based on this, the present invention provides it is a kind of include orlistat and copolymer 1a or copolymer 1 b or copolymer 1 c
The repetitive unit of nanosphere, the copolymer in the single microballoon of nanosphere is fixed with m:n ratio.
Further, the orlistat nanosphere is made by solvent evaporation method, specifically, including following step
It is rapid:
(1) orlistat and aforementioned copolymer 1a or copolymer 1 b or copolymer 1 c are dissolved in dispersed phase, ultrasonic wave is super
Sound, which sufficiently dissolves, is made oily phase；
(2) the resulting oil of step (1) is mutually injected into surfactant dropwise, stirring forms first time lotion；
(3) by the resulting first time emulsion injection of step (2) into surfactant emulsion agent, stirring is lower until organic solvent volatilizees
Completely, second of lotion is formed；
(4) resulting second of the lotion of step (3) is centrifuged, then be washed with deionized, collect microballoon, dry, sealing
It saves.
Wherein, dispersed phase described in step (1) is selected from one of ethyl alcohol, acetone, first hydrogen furans, methanol or a variety of
Mixture；Surfactant described in step (2) and step (3) is selected from polyethylene, polyvinyl alcohol, spits wet -80, gelatin, hydroxyl
One of third methylcellulose, polysorbate.
Further, the present invention optimizes each copolymer and orlistat of aforementioned preferred m:n ratio using encapsulation rate as index
Mass ratio in nanosphere, specially as follows:
For the nanosphere of copolymer 1 a for being 82:18 containing orlistat and aforementioned m:n ratio, wherein Ao Li
The mass ratio for taking charge of he and copolymer 1a is preferably 1:3.6~1:1.8, more preferably 1:3~1:2, and most preferred mass ratio is 1:
2.6。
For the nanosphere containing the copolymer 1 b that orlistat and aforementioned m:n ratio are 76:24, wherein Ao Lisi
The mass ratio of he and copolymer 1b are preferably 1:4.4~2.6, and more preferably 1:4~1:2.8, most preferred mass ratio is 1:
3.4。
For the nanosphere containing the copolymer 1 c that orlistat and aforementioned m:n ratio are 68:32, wherein Ao Lisi
The mass ratio of he and copolymer 1c are preferably 1:3.8~1:2.2, more preferably 1:3.2~1:2.6, and most preferred mass ratio is
1:3。
Compared with prior art, present invention has an advantage that
(1) provided by the invention includes that the nanosphere of orlistat and copolymer 1a~1c has good encapsulating
Rate, wherein the encapsulation rate of nanosphere provided by preferred technical solution is between 82.67% to 93.83%, hence it is evident that high
In the encapsulation rate 73.21% of control nanosphere.
(2) animal test results are shown, most preferred orlistat nanosphere orally administration rat provided by the invention
Afterwards the absorption level of each vitamin be apparently higher than orlistat and vitamin stagger the time (2 hours) oral administration vitamin absorption
Level, and be on close level with the absorption of the vitamin after orally administration vitamin independent under fasting, it can effectively make up oral Austria
The missing of liposoluble vitamin caused by Li Sita, realizes safe fat-reducing purpose.
(3) animal test results are shown, to trophism after the oral administration of most preferred nanosphere provided by the invention
The inhibiting effect of obese rat body weight increase is significantly better than the effect of the orlistat oral administration of identical equivalent.
Detailed description of the invention
Fig. 1 is to contain orlistat and the encapsulation rate of the nanosphere of copolymer 1a-2 and the relationship of two kinds of composition quality ratios
Figure.
Fig. 2 is to contain orlistat and the encapsulation rate of the nanosphere of copolymer 1b-2 and the relationship of two kinds of composition quality ratios
Figure.
Fig. 3 is to contain orlistat and the encapsulation rate of the nanosphere of copolymer 1c-2 and the relationship of two kinds of composition quality ratios
Figure.
Fig. 4 is the encapsulation rate of the nanosphere containing orlistat and PMB30W and the relational graph of two kinds of composition quality ratios.
Specific embodiment
The present invention is made combined with specific embodiments below and further being elaborated, the embodiment is served only for explaining this
Invention, is not intended to limit the scope of the present invention.Test method as used in the following examples is normal unless otherwise specified
Rule method；Used material, reagent etc., unless otherwise specified, for the reagent and material commercially obtained.
The preparation of 1 vitamin A methacrylate (compound 3a) of embodiment and structural identification
Preparation: 28.65g vitamin A (0.100mol) is taken, is placed in 500mL three-neck flask, stone is added under stirring thereto
Oily ether, until vitamin A is completely dissolved, after 5mg DCC (N, N '-dicyclohexyl diimine) is added thereto, add
The saturated oil ethereal solution of 13.21g methacrylic acid (compound 2,0.15mol) is to slowly warm up to 50 DEG C and carries out instead under stirring
It answers, and to terminal using high performance liquid chromatography tracking reaction.Vacuum distillation removes petroleum ether, freezes after obtained solid washing dry
It is dry, it obtains faint yellow solid 31.55g (0.089mol), 51~52 DEG C of fusing point, yield 89%.
Structural identification:
Compound 2:1H-NMR(CDCl3) δ (ppm): 6.69 (1H, s), 6.55 (1H, s), 2.03 (3H, s).
Vitamin A:1H-NMR(CDCl3) δ (ppm): 6.55 (1H, d), 6.54 (2H, d), 6.53 (1H, dd), 6.26 (1H,
D), 5.66 (1H, t), 4.16 (2H, d), 2.16 (3H, s), 2.14 (3H, s), 1.93 (2H, t), 1.80 (3H, s), 1.77 (2H,
M), 1.46 (2H, t), 1.08 (6H, s).
Compound 3a:1H-NMR(CDCl3) δ (ppm): 6.56 (3H, d), 6.51 (1H, dd), 6.46 (1H, s), 6.38
(1H, s), 6.27 (1H, d), 5.66 (1H, t), 4.75 (2H, d), 2.15 (3H, s), 2.14 (3H, s), 2.02 (3H, s), 1.94
(2H, t), 1.80 (3H, s), 1.78 (2H, m), 1.46 (2H, t), 1.09 (6H, s).
The preparation of 2 vitamin E methacrylate (compound 3b) of embodiment and structural identification
Preparation: 43.72g vitamin E (0.100mol) is taken, is placed in 500mL three-neck flask, second is added under stirring thereto
Ether, until vitamin E is completely dissolved, after 5mg DMAP (4-dimethylaminopyridine) is added thereto, add 13.23g methyl
The saturation diethyl ether solution of acrylic acid (compound 2,0.16mol), is to slowly warm up to 50 DEG C and is reacted under stirring, and using efficient
Liquid chromatography tracking reaction is to terminal.Vacuum distillation removes ether, is freeze-dried after obtained solid washing, obtains pale solid
45.39g (0.091mol), 42~43 DEG C of fusing point, yield 91%.
Structural identification:
Compound 2:1H-NMR(CDCl3) δ (ppm): 6.69 (1H, s), 6.55 (1H, s), 2.03 (3H, s).
Vitamin E:1H-NMR(CDCl3) δ (ppm): 2.74 (2H, t), 2.12 (3H, s), 2.11 (3H, s), 2.11 (3H,
S), 1.72 (2H, t), 1.58 (3H, m), 1.46 (3H, s), 1.40~1.16 (18H, m), 0.94 (3H, d), 0.93 (3H, d),
0.92 (3H, d), 0.90 (3H, d).
Compound 3b:1H-NMR(CDCl3) δ (ppm): 6.43 (1H, s), 6.15 (1H, s), 2.73 (2H, t), 2.12 (3H,
S), 2.09 (3H, s), 2.08 (3H, s), 2.01 (3H, s), 1.73 (2H, t), 1.59 (3H, m), 1.45 (3H, s), 1.42~
1.13 (18H, m), 0.95 (3H, d), 0.94 (3H, d), 0.91 (3H, d), 0.90 (3H, d).
3 vitamin D of embodiment2The preparation of methacrylate (compound 3c) and structural identification
Preparation: 36.69g vitamin D is taken2(0.101mol) is placed in 500mL three-neck flask, is added thereto under stirring
Ether, until vitamin E is completely dissolved, after 5mg DMAP (4-dimethylaminopyridine) is added thereto, add 13.22g first
The saturation diethyl ether solution of base acrylic acid (compound 2,0.15mol), is to slowly warm up to 50 DEG C and is reacted under stirring, and using high
Effect liquid phase chromatogram method tracking reaction is to terminal.Vacuum distillation removes ether, is freeze-dried after obtained solid washing, it is solid to obtain canescence
Body 44.15g (0.095mol), 85~87 DEG C of fusing point, yield 95%.
Structural identification:
Compound 2:1H-NMR(CDCl3) δ (ppm): 6.69 (1H, s), 6.55 (1H, s), 2.03 (3H, s).
Vitamin D2:1H-NMR(CDCl3) δ (ppm): 6.43 (1H, d), 6.23 (1H, d), 5.48 (2H, s), 5.21 (1H,
S), 5.17 (1H, s), 3.53 (1H, m), 2.35 (2H, m), 2.10 (2H, s), 2.08 (2H, t), 1.99 (1H, t), 1.85 (4H,
M), 1.63 (1H, t), 1.50 (2H, t), 1.44 (2H, m), 1.30~1.24 (3H, m), 1.18 (2H, t), 1.15 (3H, s),
0.88 (12H, d).
Compound 3c:1H-NMR(CDCl3) δ (ppm): 6.45 (1H, s), 6.44 (1H, d), 6.41 (1H, s), 6.24 (1H,
D), 5.46 (1H, s), 5.46 (1H, s), 5.18 (1H, s), 5.17 (1H, s), 3.50 (1H, m), 2.37 (2H, m), 2.13 (1H,
T), 2.07 (3H, m), 1.99 (4H, m), 1.80 (2H, m), 1.80 (2H, t), 1.59 (1H, t), 1.49~1.46 (4H, m),
1.25~1.21 (5H, m), 1.14 (3H, s), 0.93 (3H, d), 0.90 (6H, d), 0.84 (3H, d).
The preparation of 4 2- methylacryoyloxyethyl phosphocholine (compound 4, MPC) of embodiment and structural identification
Preparation:
(1) 20g HEMA and 15.6mg triethylamine are taken, is placed in 500mL three-necked bottle, 200mL anhydrous tetrahydro furan is added
Dissolve above-mentioned solid.The 100mL of lower drop 21.9g COP is stirred after acquired solution is cooled to -20 DEG C without water beetle hydrogen tetrahydrofuran solution,
It is added dropwise within 1 hour, is reacted 3 hours at -20 DEG C~-30 DEG C, be filtered to remove the three second ammonium solid of chlorination of precipitation, gained filter
Liquid vacuum distillation.50mL anhydrous ether is added into the residue after distillation, is filtered to remove the three second ammonium of a small amount of chlorination of precipitation, filters
Liquid vacuum distillation is to get the intermediate OPEMA 32.6g for being in colorless liquid.
(2) it takes 5.0g OPEMA to be dissolved in 30mL anhydrous acetonitrile to be placed in 200mL glass pressure bottle, is cooled to -20 DEG C
It is rapidly added 2mL Trimethylamine Anhydrous thereto afterwards.Pressure bottle is sealed, after being warming up to 60 DEG C of reactions 16 hours, is cooled to -20 DEG C,
The white solid of precipitation is filtered under argon atmospher, is then dried under reduced pressure to get MPC (compound 4,3.1g).
Structural identification: compound 4:1H-NMR(CDCl3) δ (ppm): 6.10 (1H, s), 5.60 (1H, s), 4.26 (4H, m),
4.05 (2H, t, J=6.1), 3.75 (2H, t, J=6.1), 3.32 (9H, s), 1.90 (3H, s) are consistent with document report.
5 vitamin A methacrylate -2- methylacryoyloxyethyl phosphocholine copolymer (copolymer 1 a) of embodiment
Preparation
(1) prepare: the molar feed ratio of compound 3a and compound 4 is 1:1, and products nr is copolymer 1 a-1
Take 3.56g vitamin A methacrylate (compound 3a) and 2.64g 2- methylacryoyloxyethyl phosphoric acid gallbladder
Alkali (compound 4, MPC), the amount of reordering methanol/tetrahydrofuran (25/75, v/v) mixed solvent, is configured to the quality of compound 3a
The solution that concentration (wt%) is 20%.After acquired solution is moved in polymerisation tube, it is molten to remove that argon gas is passed through into solution
Oxygen in liquid.Reaction tube is sealed, and is cooled down after being shaken 16 hours at 60 DEG C, to stop polymerization reaction.Reaction solution is toppled over
Into ether, the solid of precipitation is collected by filtration, drying under vacuum obtains solid 4.25g.
(2) prepare: the molar feed ratio of compound 3a and compound 4 is 4:1, and products nr is copolymer 1 a-2
Take 3.56g vitamin A methacrylate (compound 3a) and 0.66g 2- methylacryoyloxyethyl phosphoric acid gallbladder
Alkali (compound 4, MPC), the amount of reordering methanol/tetrahydrofuran (16/84, v/v) mixed solvent, is configured to the quality of compound 3a
The solution that concentration (wt%) is 20%.After acquired solution is moved in polymerisation tube, it is molten to remove that argon gas is passed through into solution
Oxygen in liquid.Reaction tube is sealed, and is cooled down after being shaken 16 hours at 60 DEG C, to stop polymerization reaction.Reaction solution is toppled over
Into normal heptane, the solid of precipitation is collected by filtration, drying under vacuum obtains solid 3.99g.
6 vitamin E methacrylate -2- methylacryoyloxyethyl phosphocholine copolymer (copolymer 1 b) of embodiment
Preparation
(1) prepare: the molar feed ratio of compound 3b and compound 4 is 1:1, and products nr is copolymer 1 b-1
Take 4.99g vitamin E methacrylate (compound 3b) and 2.64g 2- methylacryoyloxyethyl phosphoric acid gallbladder
Alkali (compound 4, MPC), the amount of reordering methanol/tetrahydrofuran (20/80, v/v) mixed solvent, is configured to the quality of compound 3b
The solution that concentration (wt%) is 20%.After acquired solution is moved in polymerisation tube, it is molten to remove that argon gas is passed through into solution
Oxygen in liquid.Reaction tube is sealed, and is cooled down after being shaken 16 hours at 60 DEG C, to stop polymerization reaction.Reaction solution is toppled over
Into ether, the solid of precipitation is collected by filtration, drying under vacuum obtains solid 6.32g.
(2) prepare: the molar feed ratio of compound 3b and compound 4 is 7:3, and products nr is copolymer 1 b-2
Take 4.99g vitamin E methacrylate (compound 3b) and 1.31g 2- methylacryoyloxyethyl phosphoric acid gallbladder
Alkali (compound 4, MPC), the amount of reordering methanol/tetrahydrofuran (12/88, v/v) mixed solvent, is configured to the quality of compound 3b
The solution that concentration (wt%) is 20%.After acquired solution is moved in polymerisation tube, it is molten to remove that argon gas is passed through into solution
Oxygen in liquid.Reaction tube is sealed, and is cooled down after being shaken 16 hours at 60 DEG C, to stop polymerization reaction.Reaction solution is toppled over
Into normal heptane, the solid of precipitation is collected by filtration, drying under vacuum obtains solid 5.45g.
7 vitamin D of embodiment2Methacrylate -2- methylacryoyloxyethyl phosphocholine copolymer (copolymer
Preparation 1c)
(1) prepare: the molar feed ratio of compound 3c and compound 4 is 1:1, and products nr is copolymer 1 c-1
Take 4.64g vitamin D2Methacrylate (compound 3c) and 2.64g 2- methylacryoyloxyethyl phosphoric acid gallbladder
Alkali (compound 4, MPC), the amount of reordering methanol/tetrahydrofuran (20/80, v/v) mixed solvent, is configured to the quality of compound 3c
The solution that concentration (wt%) is 20%.After acquired solution is moved in polymerisation tube, it is molten to remove that argon gas is passed through into solution
Oxygen in liquid.Reaction tube is sealed, and is cooled down after being shaken 16 hours at 60 DEG C, to stop polymerization reaction.Reaction solution is toppled over
Into ether, the solid of precipitation is collected by filtration, drying under vacuum obtains solid 6.11g.
(2) prepare: the molar feed ratio of compound 3c and compound 4 is 6:4, and products nr is copolymer 1 c-2
Take 4.99g vitamin D2Methacrylate (compound 3c) and 1.76g 2- methylacryoyloxyethyl phosphoric acid gallbladder
Alkali (compound 4, MPC), the amount of reordering methanol/tetrahydrofuran (14/86, v/v) mixed solvent, is configured to the quality of compound 3c
The solution that concentration (wt%) is 20%.After acquired solution is moved in polymerisation tube, it is molten to remove that argon gas is passed through into solution
Oxygen in liquid.Reaction tube is sealed, and is cooled down after being shaken 16 hours at 60 DEG C, to stop polymerization reaction.Reaction solution is toppled over
Into normal heptane, the solid of precipitation is collected by filtration, drying under vacuum obtains solid 6.18g.
The structural identification of 8 copolymer of embodiment
(1)1H-NMR(CDCl3)
1. δ: the 6.60 (d) of copolymer 1 a, 6.54 (m), 6.52 (d), 6.23 (d), 5.71 (t), 4.79 (d), 4.27
(m), 4.04 (t), 3.75 (t), 3.28 (s), 2.12 (s), 2.18 (s), 1.92 (t), 1.84 (s), 1.77 (m), 1.48 (t)
1.29 (Qiang Feng, s), 1.05 (s).Compared with compound 3a and compound 4,6.38 (1H, s, compound 3a), 6.10 (1H, s, changes
Close object 4) with absorb unimodal appearance by force at unimodal disappearance and 1.29 at 5.60 (1H, s, compounds 4), demonstrate polymerization
The generation of reaction.
2. δ: the 4.25 (m) of copolymer 1 b, 4.00 (t), 3.73 (t), 3.32 (s), 2.73 (t), 2.11 (s), 2.07
(s), 2.04 (s), 1.76 (t), 1.61 (t), 1.56 (m), 1.45 (m), 1.40 (s), 1.35 (m), 1.33 (Qiang Feng, s), 1.30
~1.26 (m), 1.24 (Qiang Feng, s), 1.22~1.15 (m), 0.95 (d), 0.90 (d), 0.89 (d), 0.87 (d).With compound
3b and compound 4 are compared, 6.43 (1H, s, compound 3b), 6.15 (1H, s, compound 3b), 6.10 (1H, s, compounds 4) with
Unimodal appearance is absorbed by force at unimodal disappearance and 1.33 and 1.24 at 5.60 (1H, s, compounds 4), and it is anti-to demonstrate polymerization
The generation answered.
3. δ: the 6.48 (d) of copolymer 1 c, 6.28 (d), 5.46 (s), 5.44 (s), 5.16 (s), 4.27~4.07 (m),
3.48 (m), 3.32 (s), 2.33~2.25 (m), 2.15 (t), 2.04 (m), 1.98~1.82 (m), 1.64 (t), 1.47~
1.44 (m), 1.34 (s), 1.24~1.17 (m), 0.90 (d), 0.88 (d), 0.84 (d).With 4 phase of compound 3b and compound
Than 6.45 (1H, s, compound 3b), 6.41 (1H, s, compound 3b), 6.10 (1H, s, compounds 4) and 5.60 (1H, s, chemical combination
Object 4) at unimodal disappearance, demonstrate the generation of polymerization reaction.
(2) weight average molecular weight (Mw), number-average molecular weight (Mn) measurement and characteristic molecular amount are distributed the measurement of (D)
Using GPC/ALC 150C type gel permeation chrommatograph, at 25 DEG C, using tetrahydrofuran as solvent, using polystyrene as
Control determines the weight average molecular weight (Mw) and number-average molecular weight (Mn) of copolymer prepared in embodiment 5~7, and following
Formula calculates molecular weight distribution (D).
The molecular weight determination of copolymer prepared by embodiment 5~7 is as shown in table 3.
3 molecular weight of copolymer measurement result of table
(3) measurement (calculating of m:n ratio) of co-monomer content
The calculation formula and calculated result of m:n ratio in copolymer structure prepared by embodiment 5~7 are as shown in table 4.
The calculation formula and calculated result of m:n in 4 copolymer structure of table
The measurement of 9 orlistat of embodiment solubility in aqueous copolymers solution
It weighs that the preparation-obtained copolymer of embodiment 5~7 is appropriate, is configured to the aqueous solution that 5mL concentration is 10mg/mL,
It is added by several times into above-mentioned aqueous solution under ultrasonic vibration orlistat (Zhong Shanwanhan pharmaceutical Co. Ltd provides, 99.8%), often
Secondary 2mg obtains sample solution until there is undissolved solid.Using the orlistat in gas chromatography measurement sample solution
Concentration, chromatographic condition are as follows.
Mobile phase: acetonitrile, phosphoric acid and water (860:0.05:140)；
Standard solution: using mobile phase as the 0.5mg/mL USP orlistat RS of solvent.Sample introduction or at 5 DEG C immediately after preparation
Lower storage；
Detector: UV 195；
Chromatographic column: 3.9-mm × 15-cm, 4- μm of filler LI；
Flow velocity: 1.0mL/min；
Sampling volume: 20 μ L；
Meanwhile system suitability is had rated using standard solution above-mentioned, it is desirable that relative standard deviation≤2%.
Solubility (S) of the orlistat in copolymer 1 a~1c in 50mg/mL aqueous solution is calculated according to the following formula
In above formula, A1The peak area of representative sample solution；A2Represent the peak area of standard solution.
The measurement result of solubility is as shown in table 5.
Solubility of 5 orlistat of table in the copolymer 50mg/mL aqueous solution that embodiment 5~7 is prepared
Embodiment 10 includes the nanosphere 1a-2 of copolymer 1 a-2 and orlistatI~IIIPreparation
(1) nanosphere 1a-2ⅠPreparation
It precisely weighs 1.20g orlistat and 3.60g copolymer 1 a-2 is dissolved in 20mL ethyl alcohol, ultrasonic echography 2 minutes
Sufficiently oily phase is made in dissolution；Above-mentioned oil is mutually injected into the 80mL polyvinyl alcohol water solution that concentration is 5mg/mL dropwise at 25 DEG C
In, 2min is stirred under 2000rpm, forms first time lotion；The 120mL that first time emulsion injection to concentration is 5mg/mL is gathered again
5 hours, formation second lotions complete to organic solvent volatilization are stirred in vinyl alcohol aqueous solution, under 500rpm；Last 4000rpm
Lower centrifugation 20 minutes, is washed with deionized 2 times.Microballoon is collected, it is micro- to be dried in vacuo 12 hours nanometers to get white at room temperature
Ball 1a-2Ⅰ, it seals at 5 DEG C and is kept in dark place, it is spare.
(2) nanosphere 1a-2ⅡPreparation
It precisely weighs 1.20g orlistat and 3.12g copolymer 1 a-2 is dissolved in 20mL ethyl alcohol, ultrasonic echography 2 minutes
Sufficiently oily phase is made in dissolution；Above-mentioned oil is mutually injected into the 80mL polyvinyl alcohol water solution that concentration is 8mg/mL dropwise at 25 DEG C
In, 3min is stirred under 2000rpm, forms first time lotion；The 120mL that first time emulsion injection to concentration is 8mg/mL is gathered again
5 hours, formation second lotions complete to organic solvent volatilization are stirred in vinyl alcohol aqueous solution, under 500rpm；Last 4000rpm
Lower centrifugation 20 minutes, is washed with deionized 2 times.Microballoon is collected, it is micro- to be dried in vacuo 12 hours nanometers to get white at room temperature
Ball 1a-2Ⅱ, it seals at 5 DEG C and is kept in dark place, it is spare.
(3) nanosphere 1a-2ⅢPreparation
It precisely weighs 1.20g orlistat and 2.40g copolymer 1 a-2 is dissolved in 20mL ethyl alcohol, ultrasonic echography 2 minutes
Sufficiently oily phase is made in dissolution；At 25 DEG C by it is above-mentioned oil mutually be injected into dropwise concentration be 10mg/mL 80mL polyvinyl alcohol it is water-soluble
In liquid, 5min is stirred under 2000rpm, forms first time lotion；It is again 10mg/mL's by first time emulsion injection to concentration
5 hours, formation second lotions complete to organic solvent volatilization are stirred in 120mL polyvinyl alcohol water solution, under 500rpm；Finally
It is centrifuged 20 minutes, is washed with deionized 2 times under 4000rpm.Microballoon is collected, is dried in vacuo 12 hours at room temperature to get white
Nanosphere 1a-2Ⅲ, it seals at 5 DEG C and is kept in dark place, it is spare.
Embodiment 11 includes the nanosphere 1b-2 of copolymer 1 b-2 and orlistatI~IIIPreparation
(1) nanosphere 1b-2ⅠPreparation
It precisely weighs 1.20g orlistat and 4.80g copolymer 1 b-2 is dissolved in 20mL acetone, ultrasonic echography 2 minutes
Sufficiently oily phase is made in dissolution；Above-mentioned oil is mutually injected into the 80mL that concentration is 2.55mg/mL dropwise at 25 DEG C, and to spit wet -80 water-soluble
In liquid, 2min is stirred under 2000rpm, forms first time lotion；It is again 2.55mg/mL's by first time emulsion injection to concentration
120mL is spat in wet -80 aqueous solution, and 5 hours, formation second lotions complete to organic solvent volatilization are stirred under 500rpm；Finally
It is centrifuged 20 minutes, is washed with deionized 2 times under 4000rpm.Microballoon is collected, is dried in vacuo 12 hours at room temperature to get white
Nanosphere 1b-2Ⅰ, it seals at 5 DEG C and is kept in dark place, it is spare.
(2) nanosphere 1b-2ⅡPreparation
It precisely weighs 1.20g orlistat and 4.08g copolymer 1 b-2 is dissolved in 20mL acetone, ultrasonic echography 2 minutes
Sufficiently oily phase is made in dissolution；Above-mentioned oil is mutually injected into the 80mL that concentration is 3.00mg/mL dropwise at 25 DEG C, and to spit wet -80 water-soluble
In liquid, 3min is stirred under 2000rpm, forms first time lotion；It is again 3.00mg/mL's by first time emulsion injection to concentration
120mL is spat in wet -80 aqueous solution, and 5 hours, formation second lotions complete to organic solvent volatilization are stirred under 500rpm；Finally
It is centrifuged 20 minutes, is washed with deionized 2 times under 4000rpm.Microballoon is collected, is dried in vacuo 12 hours at room temperature to get white
Nanosphere 1b-2Ⅱ, it seals at 5 DEG C and is kept in dark place, it is spare.
(3) nanosphere 1b-2ⅢPreparation
It precisely weighs 1.20g orlistat and 3.36g copolymer 1 b-2 is dissolved in 20mL acetone, ultrasonic echography 2 minutes
Sufficiently oily phase is made in dissolution；Above-mentioned oil is mutually injected into the 80mL that concentration is 4.00mg/mL dropwise at 25 DEG C, and to spit wet -80 water-soluble
In liquid, 5min is stirred under 2000rpm, forms first time lotion；It is again 4.00mg/mL's by first time emulsion injection to concentration
120mL is spat in wet -80 aqueous solution, and 5 hours, formation second lotions complete to organic solvent volatilization are stirred under 500rpm；Finally
It is centrifuged 20 minutes, is washed with deionized 2 times under 4000rpm.Microballoon is collected, is dried in vacuo 12 hours at room temperature to get white
Nanosphere 1b-2Ⅲ, it seals at 5 DEG C and is kept in dark place, it is spare.
Embodiment 12 includes the nanosphere 1c-2 of copolymer 1 c-2 and orlistatI~IIIPreparation
(1) nanosphere 1c-2ⅠPreparation
It precisely weighs 1.20g orlistat and 3.84g copolymer 1 c-2 is dissolved in 20mL tetrahydrofuran, ultrasonic echography 2
Oily phase is made in minute sufficiently dissolution；Above-mentioned oil is mutually injected into the 80mL hydroxypropyl first that concentration is 3.00mg/mL dropwise at 25 DEG C
In base cellulose aqueous solution, 2min is stirred under 2000rpm, forms first time lotion；It is to concentration by first time emulsion injection again
5 hours, shapes complete to organic solvent volatilization are stirred in the 120mL hypromellose aqueous solution of 2.55mg/mL, under 500rpm
At second of lotion；It is centrifuged 20 minutes, is washed with deionized 2 times under last 4000rpm.Microballoon is collected, vacuum is dry at room temperature
The dry 12 hours nanosphere 1c-2 to get whiteⅠ, it seals at 5 DEG C and is kept in dark place, it is spare.
(2) nanosphere 1c-2ⅡPreparation
It precisely weighs 1.20g orlistat and 3.60g copolymer 1 c-2 is dissolved in 20mL tetrahydrofuran, ultrasonic echography 2
Oily phase is made in minute sufficiently dissolution；Above-mentioned oil is mutually injected into the 80mL hydroxypropyl first that concentration is 3.50mg/mL dropwise at 25 DEG C
In base cellulose aqueous solution, 3min is stirred under 2000rpm, forms first time lotion；It is to concentration by first time emulsion injection again
5 hours, shapes complete to organic solvent volatilization are stirred in the 120mL hypromellose aqueous solution of 3.50mg/mL, under 500rpm
At second of lotion；It is centrifuged 20 minutes, is washed with deionized 2 times under last 4000rpm.Microballoon is collected, vacuum is dry at room temperature
The dry 12 hours nanosphere 1c-2 to get whiteⅡ, it seals at 5 DEG C and is kept in dark place, it is spare.
(3) nanosphere 1c-2ⅢPreparation
It precisely weighs 1.20g orlistat and 3.12g copolymer 1 c-2 is dissolved in 20mL tetrahydrofuran, ultrasonic echography 2
Oily phase is made in minute sufficiently dissolution；Above-mentioned oil is mutually injected into the 80mL hydroxypropyl first that concentration is 4.00mg/mL dropwise at 25 DEG C
In base cellulose aqueous solution, 5min is stirred under 2000rpm, forms first time lotion；It is to concentration by first time emulsion injection again
The 120mL of 4.00mg/mL is spat in wet -80 aqueous solution, and, formation second complete to organic solvent volatilization in 5 hours is stirred under 500rpm
Secondary lotion；It is centrifuged 20 minutes, is washed with deionized 2 times under last 4000rpm.Microballoon is collected, it is small to be dried in vacuo 12 at room temperature
When to get white nanosphere 1c-2Ⅲ, it seals at 5 DEG C and is kept in dark place, it is spare.
The preparation for compareing nanosphere of 13 orlistat of embodiment and PMB30W
PMB30W is 2- methylacryoyloxyethyl phosphocholine (MPC) and butyl methacrylate (BMA) with 7:3 moles
Than the copolymer of formation, research (Journal of of the preparation method with reference to Tomohiro Konno et al.
BiomedicalMaterial Research, 2003,65A (2): 209~214).
It precisely weighs 1.20g orlistat and 1.20g PMB30W is dissolved in 20mL tetrahydrofuran, ultrasonic echography 2 minutes
Sufficiently oily phase is made in dissolution；Above-mentioned oil is mutually injected into the 80mL hydroxypropyl fibre that concentration is 4.00mg/mL dropwise at 25 DEG C
It ties up in plain aqueous solution, 5min is stirred under 2000rpm, form first time lotion；It is to concentration by first time emulsion injection again
The 120mL of 4.00mg/mL is spat in wet -80 aqueous solution, and, formation second complete to organic solvent volatilization in 5 hours is stirred under 500rpm
Secondary lotion；It is centrifuged 20 minutes, is washed with deionized 2 times under last 4000rpm.Microballoon is collected, it is small to be dried in vacuo 12 at room temperature
When to get white control nanosphere, seal at 5 DEG C and be kept in dark place, it is spare.
Test example one, the partial size of nanosphere and encapsulation rate evaluation
(1) measurement of partial size
It is micro- that the nanometer that embodiment 10~13 is prepared is determined using JSM-5600LV type scanning electron microscope (SEM)
The partial size of ball, the results are shown in Table 6.
The partial size for the nanosphere that 6 embodiment 10~13 of table is prepared
(2) measurement of encapsulation rate
The present invention uses dialysis, and calculates the packet of the preparation-obtained nanosphere of embodiment 10~13 according to the following formula
Envelope rate, the results are shown in Table 7.
The encapsulation rate for the nanosphere that 7 embodiment 10~13 of table is prepared
The results show that the partial size of orlistat nanosphere made from 10-12 of the embodiment of the present invention is significantly less than embodiment
13 control nanospheres obtained, and there is preferable encapsulation rate, encapsulation rate is 82.67% between 93.83%, hence it is evident that high
In the encapsulation rate 73.21% of control nanosphere.
The influence evaluation of test example two, nanosphere to fat soluble vitamin absorption
(1) material:
1. drug: orlistat capsule (specification 0.12g), the production of Zhong Shanwanhan pharmaceutical Co. Ltd are used before use
0.5% sodium carboxymethylcellulose (CMC-Na) is configured to the stomach-filling liquid of 3g/L, by the dosage gastric infusion of 60mg/kg/d；
Nanosphere mixture, the nanosphere 1a-2 prepared comprising embodiment 10~12Ⅱ, nanosphere 1b-2ⅡWith receive
Meter Wei Qiu 1c-2Ⅱ, mixed by the mass ratio of 1:1:1, being configured to orlistat concentration with 0.5%CMC-Na before use is 3g/L
Stomach-filling liquid, by 60mg/kg/d (based on orlistat) given low be administered；
Vitamin mixtures include vitamin A, E, D2, mix in the ratio (W/W/W) of 1.8:2.5:2.0, use before use
0.5%CMC-Na is configured to the stomach-filling liquid that single vitamin A concentration is 2g/L, by the agent of 106mg/kg/d (based on vitamin A)
Measure gastric infusion.
2. animal and feed
SD rat, be purchased from Zhongshan University's Experimental Animal Center, 40, half male and half female, 75~85g of weight.
Normal diet is purchased from Hunan SJA Laboratory Animal Co. , Ltd, wherein containing moisture 9.7%, crude protein
20.5%, crude fat 4.62%, coarse ash 6.2%, crude fibre 4.35%, nitrogen-free extracts 50.51%, calcium 1.23%, phosphorus
0.91%, lysine 1.3%, methionine+cystine 0.68%.
(2) test method:
By SD rat, it is randomly divided into and is administered simultaneously group, administration group of staggering the time, nanosphere group and vitamin mixtures group, often
Group 10, half male and half female.
Wherein, it is administered simultaneously group, with the CMC-Na stomach-filling liquid of aforementioned orlistat and dimension while feeding with normal diet
The CMC-Na stomach-filling liquid of raw element mixture presses the dosage gastric infusion, and extracts tail vein after gastric infusion 2 hours
Liquid 0.1mL；
It staggers the time administration group, with the CMC-Na stomach-filling liquid of aforementioned orlistat by the agent while feeding with normal diet
Gastric infusion is measured, presses the dosage gastric infusion with the CMC-Na stomach-filling liquid of foregoing vitamin mixture after 2 hours, and tieing up
Raw element mixture extracts tail vein blood 0.1mL after gastric infusion 2 hours；
Nanosphere group, with the CMC-Na stomach-filling liquid of aforementioned nanosphere mixture by institute while feeding with normal diet
The dosage gastric infusion stated, and tail vein blood 0.1mL is extracted after gastric infusion 2 hours；
Vitamin mixtures group presses the dosage with the CMC-Na stomach-filling liquid of foregoing vitamin mixture under fasted conditions
Gastric infusion, and tail vein blood 0.1mL is extracted after gastric infusion 2 hours.
(3) blood is detected using high performance liquid chromatography (HPLC)
1. chromatographic condition:
Chromatographic column: octadecylsilane chemically bonded silica is filler；
Detection wavelength: 265nm；
Mobile phase and flow velocity: with methanol-water (85:15) for mobile phase A, methanol is Mobile phase B, carries out gradient elution, such as
Shown in table 8.
8 gradient elution method of table
| Time (divides) | Flow velocity (mL/ minutes) | A% |
| 0 | 2.0 | 30 |
| 17.0 | 2.0 | 30 |
| 17.5 | 3.0 | 20 |
| 34.5 | 3.0 | 20 |
| 35.0 | 3.0 | 20 |
| 35.5 | 4.0 | 0 |
| 39.5 | 4.0 | 0 |
| 40.5 | 4.0 | 30 |
| 41.0 | 2.0 | 30 |
2. for the preparation for counting blood serum sample
The tail vein blood of aforementioned each group is placed in 1mL centrifuge tube, n-hexane 1mL is separately added into, is vortexed and mixes 30 seconds,
It is centrifuged 10 minutes with being centrifuged radius 3cm, revolving speed 15000rpm.Accurate Aspirate supernatant 0.8mL, with being dried with nitrogen, residue 0.2mL
Ethyl alcohol redissolves, and 20 μ L of Aspirate supernatant after dissolution, sample introduction is analyzed.
3. each group is for vitamin A, E, D in examination blood serum sample2The comparison result of concentration is shown in Table 9.
Vitamin A, E, D in 9 each group blood serum sample HPLC map of table2Peak area comparison result (n=10)
The results show that the orlistat nanosphere 1a-2 prepared by the embodiment of the present invention 10~12Ⅱ, orlistat nanometer
Microballoon 1b-2ⅡNanosphere 1c-2 is received with orlistatⅡIn the rat blood serum sample of the nanosphere mixture group detection of composition
Fat-soluble A, vitamin E and vitamin D2Higher content is all had, shows that orlistat provided by the invention is received
Meter Wei Qiu can effectively compensate for the missing of liposoluble vitamin caused by oral orlistat, to realize safe fat-reducing purpose.
The evaluation that test example three, nanosphere act on nutritive obesity in rats loss of weight
(1) material
1. drug: orlistat capsule (specification 0.12g), the production of Zhong Shanwanhan pharmaceutical Co. Ltd are used before use
0.5% sodium carboxymethylcellulose (CMC-Na) is configured to the stomach-filling liquid of 3g/L, by the dosage gastric infusion of 60mg/kg/d；
Nanosphere, nanosphere 1a-2 prepared by Example 10~12Ⅱ, nanosphere 1b-2ⅡWith nanosphere 1c-
2Ⅱ, it is configured to the stomach-filling liquid that orlistat concentration is 3g/L with 0.5%CMC-Na respectively before use, by 60mg/kg/d (Yi Aoli
Take charge of he count) given low administration；
2. animal and feed
SD rat, be purchased from Zhongshan University's Experimental Animal Center, 60, half male and half female, 75~85g of weight.
Normal diet is purchased from Hunan SJA Laboratory Animal Co. , Ltd, constituent are as follows: moisture 9.7%, slightly
Protein 20 .5%, crude fat 4.62%, coarse ash 6.2%, crude fibre 4.35%, nitrogen-free extracts 50.51%, calcium 1.23%,
Phosphorus 0.91%, lysine 1.3%, methionine+cystine 0.68%.
Nutritional feed is purchased from Hunan SJA Laboratory Animal Co. , Ltd, constituent are as follows: moisture 8.6%,
Crude protein 18.8%, crude fat 16.2%, coarse ash 5.2%, crude fibre 3.98%, nitrogen-free extracts 42.99.%, calcium
1.24%, phosphorus 0.83%, lysine 1.38%, methionine+cystine 0.78%.
(2) method-modeling, grouping and administration
According to obesity pharmacodynamic study and evaluation method in " Herbal pharmacodynamics research and evaluation ", using pre- preventing obesity mould
Type method.Experimental animal is randomly divided into 4 groups by weight, every group 10, is used in conjunction with the mechanism of action and usage of orlistat capsule
Amount gives blank control group to be fed with normal diet, model group, orlistat control group and nanosphere mixture when every morning 9
Group feeding nutritional feed, at night 7 when take food away, it is quantitative daily (to be eaten up with most animals to food and adjust feeding daily for principle
Expect administered dose).When orlistat control group and the nanosphere group morning 9 and 3 time-division of afternoon 2 times are pressed 10m Lkg-1Use Ao Lisi
He or nanosphere CMC-Na solution stomach-filling.Blank group and the isometric 0.5%CMC-Na solution of model control group stomach-filling, continuous 7
Weigh the changes of weight situation of each group in week.
(3) result
Weight is as shown in table 10 compared with the situation of change before administration after each group rat is administered 7 weeks.
10 rat body weight situation of change of table
As can be seen from Table 10, nanosphere 1a-2 provided by the inventionⅡ、1b-2ⅡWith 1c-2ⅡTo nutrition after oral administration
Property obese rat body weight increase inhibiting effect be significantly better than identical equivalent orlistat oral administration effect.
The above is only the preferred embodiment of the present invention, it is noted that above-mentioned preferred embodiment is not construed as pair
Limitation of the invention, protection scope of the present invention should be defined by the scope defined by the claims..For the art
For those of ordinary skill, without departing from the spirit and scope of the present invention, several improvements and modifications can also be made, these change
It also should be regarded as protection scope of the present invention into retouching.
Claims (6)
1. a kind of orlistat nanosphere, which is characterized in that include copolymer and orlistat shown in following formula 1a ~ 1c
；
Copolymer shown in the formula 1a ~ 1c is vitamin A methacrylate -2- methylacryoyloxyethyl phosphocholine
Copolymer, the structural formula of the copolymer is as shown in following formula 1a:
；
Wherein, the ratio of m:n is 62:38 ~ 82:18 in copolymer structure formula shown in formula 1a；
Alternatively, copolymer shown in the formula 1a ~ 1c is vitamin E methacrylate -2- methylacryoyloxyethyl phosphorus
Sour choline copolymer, the structural formula of the copolymer is as shown in following formula 1b:
；
Wherein, the ratio of m:n is 62:38 ~ 76:24 in copolymer structure formula shown in formula 1b；
Alternatively, copolymer shown in the formula 1a ~ 1c is vitamin D2Methacrylate -2- methylacryoyloxyethyl phosphorus
Sour choline copolymer, the structural formula of the copolymer is as shown in following formula 1c:
；
Wherein, the ratio of m:n is 64:36 ~ 68:32 in copolymer structure formula shown in formula 1c.
2. orlistat nanosphere according to claim 1, which is characterized in that copolymer structure shown in the formula 1a
The ratio of m:n is 82:18 in formula, and the mass ratio of copolymer shown in the orlistat and formula 1a is 1:3.6 ~ 1:1.8.
3. orlistat nanosphere according to claim 1, which is characterized in that copolymer structure shown in the formula 1b
The ratio of m:n is 76:24 in formula, and the mass ratio of the orlistat and copolymer 1b are 1:4.4 ~ 2.6.
4. orlistat nanosphere according to claim 1, which is characterized in that copolymer knot shown in the formula 1c
The ratio of m:n is 68:32 in structure formula, and the mass ratio of the orlistat and copolymer 1c are 1:3.8 ~ 1:2.2.
5. orlistat nanosphere according to claim 1, which is characterized in that copolymer shown in the formula 1a ~ 1c
For 2- methylacryoyloxyethyl phosphocholine and it is selected from vitamin A methacrylate, vitamin E methacrylate and dimension
Raw element D2Any one methyl acrylic ester in methacrylate carries out reaction and is made.
6. -5 any orlistat nanospheres are in preparation for treating overweight and obesity drug according to claim 1
In purposes.
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