CN108524471A - Orlistat nanoparticle and its purposes in preparing obesity treating medicine - Google Patents
Orlistat nanoparticle and its purposes in preparing obesity treating medicine Download PDFInfo
- Publication number
- CN108524471A CN108524471A CN201810350124.0A CN201810350124A CN108524471A CN 108524471 A CN108524471 A CN 108524471A CN 201810350124 A CN201810350124 A CN 201810350124A CN 108524471 A CN108524471 A CN 108524471A
- Authority
- CN
- China
- Prior art keywords
- orlistat
- copolymer
- vitamin
- formula
- nanoparticle
- 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.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5005—Wall or coating material
- A61K9/5021—Organic macromolecular compounds
- A61K9/5026—Organic macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
Landscapes
- Health & Medical Sciences (AREA)
- Public Health (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Diabetes (AREA)
- Hematology (AREA)
- Obesity (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Child & Adolescent Psychology (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to pharmaceutical technology fields, more particularly to orlistat nanoparticle and its purposes in preparing obesity treating medicine.The nanoparticle includes orlistat and selected from 2 methylacryoyloxyethyl phosphocholine copolymer of vitamin A methacrylate or 2 methylacryoyloxyethyl phosphocholine copolymer of vitamin E methacrylate or vitamin D2A kind of copolymer in 2 methylacryoyloxyethyl phosphocholine copolymer of methacrylate.The encapsulation rate of the nanoparticle is 82.67~93.83%.Animal experiment is shown, after nanoparticle 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 independent orally administration vitamin under fasting, liposoluble vitamin caused by oral orlistat can effectively be made up to lack, realize safe fat-reducing purpose.
Description
Technical field
The present invention relates to pharmaceutical technology field, more particularly to a kind of orlistat nanoparticle and its fat preparing treatment
Purposes in disease drug.
Background technology
Orlistat (orlistat) is to research and develop lipase inhibitor class slimming drugs, trade name by company of Roche Group
Xenical, the last century nineties end take the lead in listing in America and Europe, are eaten by China in Discussion on Chinese Listed, and in 2005 within 2001
The approval of product Drug Administration switchs to non-prescribed medicine.Entitled N- formyls-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 to make enzyme inactivate
Triglyceride hydrolysis at absorbable aliphatic 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, Bray GA 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, there is also have many deficiencies when for treating obesity for the oral preparation of orlistat
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 is 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 that " production or processing nanometer materials manipulate 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 breast, polymer micelle, dendritic copolymer, nano medicine crystal
Deng.Oral administration nano-drug administration system has the characteristics that：1. the grain size of drug bearing microsphere, within the scope of 10~1000nm, 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 load powder is 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
System also has slow-release controlled-release, target function etc..
Andrej Dolenc et al. (International Journal of Pharmaceutics, 2010, (396) 1
~2:149~155) orlistat is scattered in Tween-80, PVP K-30, poloxamer-188 and lauryl sodium sulfate
In the aqueous solution of various stabilizers, and it is prepared for the orlistat of 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 provides orlistat in single pharmaceutical preparation and has simultaneously in the prior art fully absorbs level
Liposoluble vitamin nanometer product technical teaching.
Invention content
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 nanoparticle of safe fat-reducing purpose.
Another object of the present invention is to provide a kind of orlistat nanoparticle to prepare treatment overweight and fat
Purposes in disease drug.
In order to realize the above object, first, the present invention is synthetically prepared according to reaction route shown in figure below 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 are one of common industrial chemicals.
Compound 4 is 2- methylacryoyloxyethyls phosphocholine (MPC), can be commercial product, also can refer to
Ishihara K et al.(Polym J 1990；22:355-360.) with Ueda T et al. (Polym J 1992；24:
1259-1269.) method disclosed by is prepared.
ROH is selected from VitAVitE or vitamin D2In a kind 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 be：N, N '-dicyclohexyl diimine (DCC) or 4- diformazans
Aminopyridine (DMAP), petroleum ether, 50~60 DEG C.The step is conventional acylation reaction, operation can according to but be not limited to hear
Tough chief editor's《Drug synthetic reaction》What (Chemical Industry Press publishes in April, 2017) and Liu Xianghong et al. were delivered《One-step method
Synthesize Retinol Palmitate》The method that (Chemical Industry in Guangzhou, the 14th phase of volume 39 in 2011,63~64) discloses carries out.Described
Methacrylic acid：The molar ratio of ROH is 1:1~2:Between 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 be：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 the copolymer 1 a~1c being prepared.
Specifically, compound 3a~3c and the rate of charge confirmation method of compound 4 are in above-mentioned steps b：With 1：1 conductization
Close the minimum molar feed ratio of each compound and compound 4 in object 3a~3c, and the dissolving 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 formulas 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 formulas 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 with product structure has been carried out to the aforementioned raw material per single step reaction
Card, and by comparing product and raw material1The similarities and differences of H-NMR spectrum datas, to conclude the generation of reaction.
(2) m in copolymer 1 a~1c structural formulas:N than calculating
The present invention is by calculating copolymer 1 a~1c's1- N in H-NMR collection of illustrative plates+(CH3)3The peak area of the characteristic peak of hydrogen accounts for
The ratio of total hydrogen peak area, calculates the m in copolymer 1 a~1c structural formulas:N ratios.
(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,
As a contrast with polystyrene, the various m of aforementioned gained are determined:N than copolymer 1 a~1c molecular weight.
Further, the structural formula of the copolymer 1 a~1c is as shown below：
Wherein, if the copolymer is copolymer 1 a, the m in structural formula:N ratios are 62:38~82:It is excellent between 18
It is selected as 82:18；
If the copolymer is copolymer 1 b, the m in structural formula:N ratios are 62:38~76:Between 24, preferably
76:24；
If the copolymer is copolymer 1 c, the m in structural formula:N ratios are 64:36~68:Between 32, preferably
68:32。
Further, of the invention in order to verify whether copolymer 1 a~1c has humidification to orlistat water solubility
People has carried out solubility test of the orlistat in copolymer 1 a~1c aqueous solutions, specific to use《Chinese Pharmacopoeia》(2015 editions)
Specified in method determine solubility of the orlistat in the 50mg/mL aqueous solutions of aforementioned copolymer 1a~1c, particularly,
The present invention determine orlistat it is maximum in the 50mg/mL aqueous solutions of 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
Nanoparticle, the repetitive unit and m of the copolymer in the single microballoon of nanoparticle:N ratios are fixed.
Further, the orlistat nanoparticle is made by solvent evaporation method, specifically, including following step
Suddenly：
(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 fully dissolves, is made oil phase；
(2) oil phase obtained by step (1) is injected into surfactant dropwise, stirring forms first time lotion；
(3) by the first time emulsion injection to surfactant emulsion agent obtained by step (2), stirring is lower until organic solvent volatilizees
Completely, second of lotion is formed；
(4) second of lotion obtained by step (3) is centrifuged, then be washed with deionized, collect microballoon, dry, sealing
It preserves.
Wherein, dispersed phase described in step (1) is one or more in ethyl alcohol, acetone, first hydrogen furans, methanol
Mixture；Step (2) and surfactant described in step (3) are selected from polyethylene, polyvinyl alcohol, spit wet -80, gelatin, hydroxyl
One kind in third methylcellulose, polysorbate.
Further, the present invention optimizes aforementioned preferred m using encapsulation rate as index:N than each copolymer and orlistat
Mass ratio in nanoparticle, it is specially as follows：
For containing orlistat and aforementioned m:N ratios are 82:For the nanoparticle of 18 copolymer 1 a, wherein Ao Li
The mass ratio that him is taken charge of with copolymer 1a is preferably 1:3.6~1:1.8, more preferably 1:3~1:2, most preferred mass ratio is 1:
2.6。
For containing orlistat and aforementioned m:N ratios are 76:For the nanoparticle of 24 copolymer 1 b, wherein Ao Lisi
The mass ratio of he and copolymer 1b are preferably 1:4.4~2.6, more preferably 1:4~1:2.8, most preferred mass ratio is 1:
3.4。
For containing orlistat and aforementioned m:N ratios are 68:For the nanoparticle of 32 copolymer 1 c, 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, most preferred mass ratio is
1:3。
Compared with prior art, advantage of the invention is that：
(1) provided by the invention includes orlistat and the nanoparticle of copolymer 1a~1c has good encapsulating
Rate, wherein the encapsulation rate for the nanoparticle that preferred technical solution is provided is between 82.67% to 93.83%, hence it is evident that high
In the encapsulation rate 73.21% of control nanoparticle.
(2) animal test results are shown, most preferred orlistat nanoparticle 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 independent orally administration vitamin under fasting, it can effectively make up oral Austria
Liposoluble vitamin caused by Li Sita lacks, and realizes safe fat-reducing purpose.
(3) animal test results are shown, to trophism after the oral medication of most preferred nanoparticle provided by the invention
The inhibiting effect of obese rat body weight increase is significantly better than the effect of the orlistat oral medication of identical equivalent.
Description of the drawings
Fig. 1 be the encapsulation rate containing orlistat and the nanoparticle of copolymer 1a-2 and two kinds of composition qualities than relationship
Figure.
Fig. 2 be the encapsulation rate containing orlistat and the nanoparticle of copolymer 1b-2 and two kinds of composition qualities than relationship
Figure.
Fig. 3 be the encapsulation rate containing orlistat and the nanoparticle of copolymer 1c-2 and two kinds of composition qualities than relationship
Figure.
Fig. 4 be the nanoparticle containing orlistat and PMB30W encapsulation rate and two kinds of composition qualities than relational graph.
Specific implementation mode
The present invention is made with reference to specific embodiment 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 used in following embodiments 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
It prepares：28.65g vitamin As (0.100mol) are taken, is placed in 500mL three-neck flasks, stone is added under stirring thereto
Oily ether, until vitamin A is completely dissolved, after 5mg DCC (N, N '-dicyclohexyl diimine) are added thereto, add
The saturated oil ethereal solution of 13.21g methacrylic acids (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, is freezed after obtained solid washing dry
It is dry, obtain 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
It prepares：43.72g vitamin Es (0.100mol) are taken, is placed in 500mL three-neck flasks, second is added under stirring thereto
Ether, until vitamin E is completely dissolved, after 5mg DMAP (4-dimethylaminopyridine) are 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
It prepares：Take 36.69g vitamin Ds2(0.101mol) is placed in 500mL three-neck flasks, is added thereto under stirring
Ether, until vitamin E is completely dissolved, after 5mg DMAP (4-dimethylaminopyridine) are 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- methylacryoyloxyethyls phosphocholine (compound 4, MPC) of embodiment and structural identification
It prepares：
(1) 20g HEMA and 15.6mg triethylamines are taken, is placed in 500mL three-necked bottles, 200mL anhydrous tetrahydro furans are added
Dissolve above-mentioned solid.Acquired solution is cooled to and stirs the 100mL of lower drop 21.9g COP after -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 is evaporated under reduced pressure.50mL anhydrous ethers are added into the residue after distillation, are filtered to remove the three second ammonium of a small amount of chlorination of precipitation, filter
Liquid is evaporated under reduced pressure to get in the intermediate OPEMA 32.6g of colorless liquid.
(2) it takes 5.0g OPEMA to be dissolved in 30mL anhydrous acetonitriles to be placed in 200mL glass pressure bottles, is cooled to -20 DEG C
It is rapidly added 2mL Trimethylamine Anhydrous thereto afterwards.Seal pressure bottle, be warming up to 60 DEG C reaction 16 hours after, be 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 copolymers (copolymer 1 a) of embodiment
Preparation
(1) it prepares：Compound 3a and the molar feed ratio of compound 4 are 1:1, products nr is copolymer 1 a-1
Take 3.56g vitamin A methacrylates (compound 3a) and 2.64g 2- methylacryoyloxyethyl phosphoric acid courages
Alkali (compound 4, MPC), the mixed solvent of the amount of reordering methanol/tetrahydrofuran (25/75, v/v), 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 shakes 16 hours postcoolings at 60 DEG C, to stop polymerisation.Reaction solution is toppled over
Into ether, the solid of precipitation is collected by filtration, is dried under vacuum, obtains solid 4.25g.
(2) it prepares：Compound 3a and the molar feed ratio of compound 4 are 4:1, products nr is copolymer 1 a-2
Take 3.56g vitamin A methacrylates (compound 3a) and 0.66g 2- methylacryoyloxyethyl phosphoric acid courages
Alkali (compound 4, MPC), the mixed solvent of the amount of reordering methanol/tetrahydrofuran (16/84, v/v), 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 shakes 16 hours postcoolings at 60 DEG C, to stop polymerisation.Reaction solution is toppled over
Into normal heptane, the solid of precipitation is collected by filtration, is dried under vacuum, obtains solid 3.99g.
6 vitamin E methacrylate -2- methylacryoyloxyethyl phosphocholine copolymers (copolymer 1 b) of embodiment
Preparation
(1) it prepares：Compound 3b and the molar feed ratio of compound 4 are 1:1, products nr is copolymer 1 b-1
Take 4.99g vitamin E methacrylates (compound 3b) and 2.64g 2- methylacryoyloxyethyl phosphoric acid courages
Alkali (compound 4, MPC), the mixed solvent of the amount of reordering methanol/tetrahydrofuran (20/80, v/v), 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 shakes 16 hours postcoolings at 60 DEG C, to stop polymerisation.Reaction solution is toppled over
Into ether, the solid of precipitation is collected by filtration, is dried under vacuum, obtains solid 6.32g.
(2) it prepares：Compound 3b and the molar feed ratio of compound 4 are 7:3, products nr is copolymer 1 b-2
Take 4.99g vitamin E methacrylates (compound 3b) and 1.31g 2- methylacryoyloxyethyl phosphoric acid courages
Alkali (compound 4, MPC), the mixed solvent of the amount of reordering methanol/tetrahydrofuran (12/88, v/v), 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 shakes 16 hours postcoolings at 60 DEG C, to stop polymerisation.Reaction solution is toppled over
Into normal heptane, the solid of precipitation is collected by filtration, is dried under vacuum, obtains solid 5.45g.
7 vitamin D of embodiment2Methacrylate -2- methylacryoyloxyethyl phosphocholine copolymer (copolymers
Preparation 1c)
(1) it prepares：Compound 3c and the molar feed ratio of compound 4 are 1:1, products nr is copolymer 1 c-1
Take 4.64g vitamin Ds2Methacrylate (compound 3c) and 2.64g 2- methylacryoyloxyethyl phosphoric acid courages
Alkali (compound 4, MPC), the mixed solvent of the amount of reordering methanol/tetrahydrofuran (20/80, v/v), 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 shakes 16 hours postcoolings at 60 DEG C, to stop polymerisation.Reaction solution is toppled over
Into ether, the solid of precipitation is collected by filtration, is dried under vacuum, obtains solid 6.11g.
(2) it prepares：Compound 3c and the molar feed ratio of compound 4 are 6:4, products nr is copolymer 1 c-2
Take 4.99g vitamin Ds2Methacrylate (compound 3c) and 1.76g 2- methylacryoyloxyethyl phosphoric acid courages
Alkali (compound 4, MPC), the mixed solvent of the amount of reordering methanol/tetrahydrofuran (14/86, v/v), 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 shakes 16 hours postcoolings at 60 DEG C, to stop polymerisation.Reaction solution is toppled over
Into normal heptane, the solid of precipitation is collected by filtration, is dried under vacuum, obtains solid 6.18g.
The structural identification of 8 copolymer of embodiment
(1)1H-NMR(CDCl3)
1. the δ of copolymer 1 a：6.60 (d), 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 (strong peak, 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. the δ of copolymer 1 b：4.25 (m), 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 (strong peak, s), 1.30
~1.26 (m), 1.24 (strong peak, 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. the δ of copolymer 1 c：6.48 (d), 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 polymerisation.
(2) weight average molecular weight (Mw), number-average molecular weight (Mn) measure the measurement with characteristic molecular amount distribution (D)
Using GPC/ALC 150C type gel permeation chrommatographs, 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 (m of co-monomer content:N than calculating)
The m in copolymer structure prepared by embodiment 5~7:N than calculation formula and result of calculation it is as shown in table 4.
M in 4 copolymer structure of table:The calculation formula and result of calculation of n
The measurement of 9 orlistat of embodiment solubility in aqueous copolymers solution
It is appropriate to weigh 5~7 preparation-obtained copolymer of embodiment, is configured to the aqueous solution of a concentration of 10mg/mL of 5mL,
Orlistat (Zhong Shanwanhan pharmaceutical Co. Ltds provide, 99.8%) is added under ultrasonic vibration into above-mentioned aqueous solution by several times, often
Secondary 2mg obtains sample solution until there is undissolved solid.Using the orlistat in gas chromatography determination 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 orlistats 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：20μL；
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 solutions 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 solutions that embodiment 5~7 is prepared
Embodiment 10 includes the nanoparticle 1a-2 of copolymer 1 a-2 and orlistatI~IIIPreparation
(1) nanoparticle 1a-2ⅠPreparation
It precisely weighs 1.20g orlistats to be dissolved in 20mL ethyl alcohol with 3.60g copolymer 1s a-2, ultrasonic echography 2 minutes
Fully oil phase is made in dissolving；Above-mentioned oil phase is injected into the 80mL polyvinyl alcohol water solutions of a concentration of 5mg/mL dropwise at 25 DEG C
In, 2min is stirred under 2000rpm, forms first time lotion；The 120mL of first time emulsion injection to a concentration of 5mg/mL are 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) nanoparticle 1a-2ⅡPreparation
It precisely weighs 1.20g orlistats to be dissolved in 20mL ethyl alcohol with 3.12g copolymer 1s a-2, ultrasonic echography 2 minutes
Fully oil phase is made in dissolving；Above-mentioned oil phase is injected into the 80mL polyvinyl alcohol water solutions of a concentration of 8mg/mL dropwise at 25 DEG C
In, 3min is stirred under 2000rpm, forms first time lotion；The 120mL of first time emulsion injection to a concentration of 8mg/mL are 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) nanoparticle 1a-2ⅢPreparation
It precisely weighs 1.20g orlistats to be dissolved in 20mL ethyl alcohol with 2.40g copolymer 1s a-2, ultrasonic echography 2 minutes
Fully oil phase is made in dissolving；The 80mL polyvinyl alcohol that above-mentioned oil phase is injected into a concentration of 10mg/mL dropwise at 25 DEG C is water-soluble
In liquid, 5min is stirred under 2000rpm, forms first time lotion；Again by first time emulsion injection to a concentration of 10mg/mL's
5 hours, formation second lotions complete to organic solvent volatilization are stirred in 120mL polyvinyl alcohol water solutions, under 500rpm；Finally
It centrifuges 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
Nanoparticle 1a-2Ⅲ, it seals at 5 DEG C and is kept in dark place, it is spare.
Embodiment 11 includes the nanoparticle 1b-2 of copolymer 1 b-2 and orlistatI~IIIPreparation
(1) nanoparticle 1b-2ⅠPreparation
It precisely weighs 1.20g orlistats to be dissolved in 20mL acetone with 4.80g copolymer 1s b-2, ultrasonic echography 2 minutes
Fully oil phase is made in dissolving；Above-mentioned oil phase is injected into the 80mL of a concentration of 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；Again by first time emulsion injection to a concentration of 2.55mg/mL's
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 centrifuges 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
Nanoparticle 1b-2Ⅰ, it seals at 5 DEG C and is kept in dark place, it is spare.
(2) nanoparticle 1b-2ⅡPreparation
It precisely weighs 1.20g orlistats to be dissolved in 20mL acetone with 4.08g copolymer 1s b-2, ultrasonic echography 2 minutes
Fully oil phase is made in dissolving；Above-mentioned oil phase is injected into the 80mL of a concentration of 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；Again by first time emulsion injection to a concentration of 3.00mg/mL's
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 centrifuges 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
Nanoparticle 1b-2Ⅱ, it seals at 5 DEG C and is kept in dark place, it is spare.
(3) nanoparticle 1b-2ⅢPreparation
It precisely weighs 1.20g orlistats to be dissolved in 20mL acetone with 3.36g copolymer 1s b-2, ultrasonic echography 2 minutes
Fully oil phase is made in dissolving；Above-mentioned oil phase is injected into the 80mL of a concentration of 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；Again by first time emulsion injection to a concentration of 4.00mg/mL's
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 centrifuges 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
Nanoparticle 1b-2Ⅲ, it seals at 5 DEG C and is kept in dark place, it is spare.
Embodiment 12 includes the nanoparticle 1c-2 of copolymer 1 c-2 and orlistatI~IIIPreparation
(1) nanoparticle 1c-2ⅠPreparation
It precisely weighs 1.20g orlistats to be dissolved in 20mL tetrahydrofurans with 3.84g copolymer 1s c-2, ultrasonic echography 2
Oil phase is made in minute fully dissolving；Above-mentioned oil phase is injected into the 80mL hydroxypropyl first of a concentration of 3.00mg/mL dropwise at 25 DEG C
In base cellulose aqueous solution, 2min is stirred under 2000rpm, forms first time lotion；Again by first time emulsion injection to a concentration of
5 hours, shapes complete to organic solvent volatilization are stirred in the 120mL hypromellose aqueous solutions of 2.55mg/mL, under 500rpm
At second of lotion；It centrifuges 20 minutes, is washed with deionized 2 times under last 4000rpm.Microballoon is collected, vacuum is dry at room temperature
The dry 12 hours nanoparticle 1c-2 to get whiteⅠ, it seals at 5 DEG C and is kept in dark place, it is spare.
(2) nanoparticle 1c-2ⅡPreparation
It precisely weighs 1.20g orlistats to be dissolved in 20mL tetrahydrofurans with 3.60g copolymer 1s c-2, ultrasonic echography 2
Oil phase is made in minute fully dissolving；Above-mentioned oil phase is injected into the 80mL hydroxypropyl first of a concentration of 3.50mg/mL dropwise at 25 DEG C
In base cellulose aqueous solution, 3min is stirred under 2000rpm, forms first time lotion；Again by first time emulsion injection to a concentration of
5 hours, shapes complete to organic solvent volatilization are stirred in the 120mL hypromellose aqueous solutions of 3.50mg/mL, under 500rpm
At second of lotion；It centrifuges 20 minutes, is washed with deionized 2 times under last 4000rpm.Microballoon is collected, vacuum is dry at room temperature
The dry 12 hours nanoparticle 1c-2 to get whiteⅡ, it seals at 5 DEG C and is kept in dark place, it is spare.
(3) nanoparticle 1c-2ⅢPreparation
It precisely weighs 1.20g orlistats to be dissolved in 20mL tetrahydrofurans with 3.12g copolymer 1s c-2, ultrasonic echography 2
Oil phase is made in minute fully dissolving；Above-mentioned oil phase is injected into the 80mL hydroxypropyl first of a concentration of 4.00mg/mL dropwise at 25 DEG C
In base cellulose aqueous solution, 5min is stirred under 2000rpm, forms first time lotion；Again by first time emulsion injection to a concentration of
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 centrifuges 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 nanoparticle 1c-2Ⅲ, it seals at 5 DEG C and is kept in dark place, it is spare.
The preparation for compareing nanoparticle of 13 orlistat of embodiment and PMB30W
PMB30W is 2- methylacryoyloxyethyls 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 orlistats to be dissolved in 20mL tetrahydrofurans with 1.20g PMB30W, ultrasonic echography 2 minutes
Fully oil phase is made in dissolving；The 80mL hydroxypropyls that above-mentioned oil phase is injected into a concentration of 4.00mg/mL dropwise at 25 DEG C are fine
In the plain aqueous solution of dimension, 5min is stirred under 2000rpm, forms first time lotion；Again by first time emulsion injection to a concentration of
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 centrifuges 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 nanoparticle, seal at 5 DEG C and be kept in dark place, it is spare.
Test example one, the grain size of nanoparticle and encapsulation rate evaluation
(1) measurement of grain size
It is micro- that the nanometer that embodiment 10~13 is prepared is determined using JSM-5600LV types scanning electron microscope (SEM)
The grain size of ball, the results are shown in Table 6.
The grain size for the nanoparticle that 6 embodiment 10~13 of table is prepared
(2) measurement of encapsulation rate
The present invention uses dialysis, and calculates the packet of 10~13 preparation-obtained nanoparticle of embodiment according to the following formula
Envelope rate, the results are shown in Table 7.
The encapsulation rate for the nanoparticle that 7 embodiment 10~13 of table is prepared
The results show that the grain size of orlistat nanoparticle made from 10-12 of the embodiment of the present invention is significantly less than embodiment
13 control nanoparticles 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 nanoparticle.
The influence evaluation of test example two, nanoparticle to fat soluble vitamin absorption
(1) material：
1. drug：Orlistat capsule (specification 0.12g), Zhong Shanwanhan pharmaceutical Co. Ltds production, is used before use
0.5% sodium carboxymethylcellulose (CMC-Na) is configured to the gavage liquid of 3g/L, by the dosage gastric infusion of 60mg/kg/d；
Nanoparticle mixture, including nanoparticle 1a-2 prepared by embodiment 10~12Ⅱ, nanoparticle 1b-2ⅡWith receive
Meter Wei Qiu 1c-2Ⅱ, by 1:1:1 mass ratio mixing, is configured to a concentration of 3g/L of orlistat with 0.5%CMC-Na before use
Gavage liquid, by 60mg/kg/d (based on orlistat) given low be administered；
Vitamin mixtures, including vitamin A, E, D2, by 1.8:2.5:2.0 ratio (W/W/W) mixing, is used before use
0.5%CMC-Na is configured to the gavage 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 rats, 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 rats, it is randomly divided into and is administered simultaneously group, administration group of staggering the time, nanoparticle group and vitamin mixtures group, often
Group 10, half male and half female.
Wherein, it is administered simultaneously group, with the CMC-Na gavages liquid of aforementioned orlistat and dimension while feeding with normal diet
The CMC-Na gavage liquid of raw element mixture presses the dosage gastric infusion, and extracts tail vein after 2 hours in gastric infusion
Liquid 0.1mL；
It staggers the time administration group, with the CMC-Na gavages 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 gavage liquid of foregoing vitamin mixture after 2 hours, and tieing up
Raw element mixture gastric infusion extracts tail vein blood 0.1mL after 2 hours；
Nanoparticle group, with the CMC-Na gavages liquid of aforementioned nanoparticle 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 under fasted conditions with the CMC-Na gavage liquid of foregoing vitamin mixture
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) it is 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 counting the preparation of blood serum sample
The tail vein blood of aforementioned each group is placed in 1mL centrifuge tubes, is separately added into n-hexane 1mL, vortex mixing 30 seconds,
To centrifuge radius 3cm, rotating speed 15000rpm is centrifuged 10 minutes.Accurate Aspirate supernatant 0.8mL, is dried up with nitrogen, residue 0.2mL
Ethyl alcohol redissolves, and 20 μ L of Aspirate supernatant after dissolving, 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 collection of illustrative plates of table2Peak area comparison result (n=10)
The results show that the orlistat nanoparticle 1a-2 prepared by the embodiment of the present invention 10~12Ⅱ, orlistat nanometer
Microballoon 1b-2ⅡNanoparticle 1c-2 is received with orlistatⅡIn the rat blood serum sample of the nanoparticle 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 liposoluble vitamin caused by oral orlistat and lack, to realize safe fat-reducing purpose.
The evaluation that test example three, nanoparticle act on nutritive obesity in rats loss of weight
(1) material
1. drug：Orlistat capsule (specification 0.12g), Zhong Shanwanhan pharmaceutical Co. Ltds production, is used before use
0.5% sodium carboxymethylcellulose (CMC-Na) is configured to the gavage liquid of 3g/L, by the dosage gastric infusion of 60mg/kg/d；
Nanoparticle, nanoparticle 1a-2 prepared by Example 10~12Ⅱ, nanoparticle 1b-2ⅡWith nanoparticle 1c-
2Ⅱ, the gavage liquid of a concentration of 3g/L of orlistat is configured to 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 rats, 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, and constituent is：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, and constituent is：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
Foundation《Herbal pharmacodynamics are studied and evaluation》Middle obesity pharmacodynamic study and evaluation method, 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 nanoparticle 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 nanoparticle group morning 9 and 3 time-division of afternoon presses 10m Lkg 2 times-1Use Ao Lisi
He or nanoparticle CMC-Na solution gavages.Blank group and the isometric 0.5%CMC-Na solution of model control group gavage, 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, nanoparticle 1a-2 provided by the inventionⅡ、1b-2ⅡWith 1c-2ⅡTo nutrition after oral medication
Property obese rat body weight increase inhibiting effect be significantly better than identical equivalent orlistat oral medication effect.
It the above is only the preferred embodiment of the present invention, it is noted that above-mentioned preferred embodiment is not construed as pair
The limitation of the present invention, protection scope of the present invention should be subject to claim limited range.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
Protection scope of the present invention is also should be regarded as into retouching.
Claims (10)
1. a kind of orlistat nanoparticle, which is characterized in that include copolymer and orlistat shown in following formula 1a~1c
2. orlistat nanoparticle according to claim 1, which is characterized in that be copolymerized shown in the formula 1a~1c
Substituent R in object structural formula is：
3. orlistat nanoparticle according to claim 1 or 2, which is characterized in that shown in the formula 1a~1c
Copolymer is vitamin A methacrylate -2- methylacryoyloxyethyl phosphocholine copolymers, the structure of the copolymer
Formula is as shown in following formula 1a：
Wherein, m in copolymer structure formula shown in formula 1a:The ratio of n is 62:38~82:18.
4. orlistat nanoparticle according to claim 3, which is characterized in that copolymer structure shown in the formula 1a
M in formula:The ratio of n is 82:18, the mass ratio of copolymer shown in the orlistat and formula 1a is 1:3.6~1:1.8.
5. orlistat nanoparticle according to claim 1 or 2, which is characterized in that shown in the formula 1a~1c
Copolymer is vitamin E methacrylate -2- methylacryoyloxyethyl phosphocholine copolymers, the structure of the copolymer
Formula is as shown in following formula 1b：
Wherein, m in copolymer structure formula shown in formula 1b:The ratio of n is 62:38~76:24.
6. orlistat nanoparticle according to claim 5, which is characterized in that copolymer structure shown in the formula 1b
M in formula:The ratio of n is 76:24, the mass ratio of the orlistat and copolymer 1b are 1:4.4~2.6.
7. orlistat nanoparticle according to claim 1 or 2, which is characterized in that shown in the formula 1a~1c
Copolymer is vitamin D2Methacrylate -2- methylacryoyloxyethyl phosphocholine copolymers, the knot of the copolymer
Structure formula is as shown in following formula 1c：
Wherein, m in copolymer structure formula shown in formula 1c:The ratio of n is 64:36~68:32.
8. orlistat nanoparticle according to claim 7, which is characterized in that copolymer structure shown in the formula 1c
M in formula:The ratio of n is 68:32, the mass ratio of the orlistat and copolymer 1c are 1:3.8~1:2.2.
9. orlistat nanoparticle according to claim 1, which is characterized in that copolymer shown in the formula 1a~1c
For 2- methylacryoyloxyethyls 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.
10. being prepared according to any orlistat nanoparticles of claim 1-9 for treating overweight and obesity medicine
Purposes in object.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810350124.0A CN108524471B (en) | 2018-04-18 | 2018-04-18 | Orlistat nanosphere and its preparing the purposes in obesity treating medicine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810350124.0A CN108524471B (en) | 2018-04-18 | 2018-04-18 | Orlistat nanosphere and its preparing the purposes in obesity treating medicine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108524471A true CN108524471A (en) | 2018-09-14 |
| CN108524471B CN108524471B (en) | 2019-03-01 |
Family
ID=63477785
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810350124.0A Active CN108524471B (en) | 2018-04-18 | 2018-04-18 | Orlistat nanosphere and its preparing the purposes in obesity treating medicine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108524471B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109157523A (en) * | 2018-10-09 | 2019-01-08 | 中山万汉制药有限公司 | A kind of orlistat dripping pill and preparation method thereof |
| WO2021097652A1 (en) * | 2019-11-19 | 2021-05-27 | 中山万汉制药有限公司 | Orlistat nanospheres and use thereof in preparation of medicines for treatment of obesity |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1180013B1 (en) * | 1999-05-27 | 2006-03-08 | Biocompatibles UK Limited | Local drug delivery |
| CN102348468A (en) * | 2009-07-31 | 2012-02-08 | 西安力邦医药科技有限责任公司 | Nanosphere or microsphere drug carrier, preparation method, composition and use thereof |
| CN106562933A (en) * | 2016-11-10 | 2017-04-19 | 广西大学 | Preparation method for lignin drug sustained release microspheres |
| CN107412196A (en) * | 2017-05-08 | 2017-12-01 | 重庆植恩药业有限公司 | Orlistat nanoparticle and preparation method thereof and the application in antineoplastic |
-
2018
- 2018-04-18 CN CN201810350124.0A patent/CN108524471B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1180013B1 (en) * | 1999-05-27 | 2006-03-08 | Biocompatibles UK Limited | Local drug delivery |
| CN102348468A (en) * | 2009-07-31 | 2012-02-08 | 西安力邦医药科技有限责任公司 | Nanosphere or microsphere drug carrier, preparation method, composition and use thereof |
| CN106562933A (en) * | 2016-11-10 | 2017-04-19 | 广西大学 | Preparation method for lignin drug sustained release microspheres |
| CN107412196A (en) * | 2017-05-08 | 2017-12-01 | 重庆植恩药业有限公司 | Orlistat nanoparticle and preparation method thereof and the application in antineoplastic |
Non-Patent Citations (6)
| Title |
|---|
| FATHIMA A等: "MICROPARTICULATE DRUG DELIVERY SYSTEM OF ORLISTAT FOR SUSTAINED RELEASE", 《INT. J. PHARM. SCI. REV. RES.》 * |
| KAZUHIKO ISHIHARA等: "Polymeric Lipid Nanosphere Consisting of Water-soluble Poly(2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate)", 《POLYMER JOURNAL》 * |
| RAQUEL PALAO-SUAY等: "Anticancer and Antiangiogenic Activity of Surfactant-Free Nanoparticles Based on Self-Assembled Polymeric Derivatives of Vitamin E: Structure−Activity Relationship", 《BIOMACROMOLECULES》 * |
| S. B. SATEESHA等: "Gastroretentive Orlistat Microspheres: Formulation, Characterization, and In Vitro Evaluation", 《DISSOLUTION TECHNOLOGIES》 * |
| SIRONG TAN等: "Antiobese Effects of Capsaicin−Chitosan Microsphere (CCMS) in Obese Rats Induced by High Fat Diet", 《J. AGRIC. FOOD CHEM.》 * |
| TOMOHIRO KONNO等: "Enhanced solubility of paclitaxel using water-soluble and biocompatible 2-methacryloyloxyethyl phosphorylcholine polymers", 《J BIOMED MASTER RES A》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109157523A (en) * | 2018-10-09 | 2019-01-08 | 中山万汉制药有限公司 | A kind of orlistat dripping pill and preparation method thereof |
| CN109157523B (en) * | 2018-10-09 | 2020-07-28 | 中山万汉制药有限公司 | Orlistat dripping pill and its prepn |
| WO2021097652A1 (en) * | 2019-11-19 | 2021-05-27 | 中山万汉制药有限公司 | Orlistat nanospheres and use thereof in preparation of medicines for treatment of obesity |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108524471B (en) | 2019-03-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Muley et al. | Hydrophobically modified inulin as an amphiphilic carbohydrate polymer for micellar delivery of paclitaxel for intravenous route | |
| Zhang et al. | Solubility and bioavailability enhancement of oridonin: a review | |
| KR100517253B1 (en) | pH responsive biodegradable polylactic acid forming polymeric micelle and their use for poorly water soluble drug delivery | |
| Park et al. | Comparison of a revaprazan-loaded solid dispersion, solid SNEDDS and inclusion compound: physicochemical characterisation and pharmacokinetics | |
| CN108524471B (en) | Orlistat nanosphere and its preparing the purposes in obesity treating medicine | |
| Qin et al. | PEGylated solanesol for oral delivery of coenzyme Q10 | |
| CN104961886B (en) | A kind of preparation method of nano-micelle medicinal materials | |
| CN106727336B (en) | Oridonin cubic liquid crystal nanoparticles and preparation method thereof | |
| CN108175860B (en) | Esterase response type curcumin-polymerized thiodipropionic acid copolymer prodrug nano-micelle and preparation method and application thereof | |
| Karan et al. | Polymeric microsphere formulation for Colon targeted delivery of 5-fluorouracil using biocompatible natural gum Katira | |
| Shao et al. | Development and evaluation of self-microemulsifying liquid and granule formulations of Brucea javanica oil | |
| CN106727309A (en) | Polymer micelle solution containing Quercetin and its preparation method and application | |
| Chen et al. | Modifying glycyrrhetinic acid liposomes with liver-targeting ligand of galactosylated derivative: preparation and evaluations | |
| Zhang et al. | Hollow microcapsules with ulcerative colitis therapeutic effects made of multifunctional Turkish galls extraction | |
| CN108578390B (en) | The purposes of nanosphere containing orlistat in the preparation of antitumor drugs | |
| Thanos et al. | Improving relative bioavailability of dicumarol by reducing particle size and adding the adhesive poly (fumaric-co-sebacic) anhydride | |
| Zou et al. | Andrographolide/phospholipid/cyclodextrin complex-loaded nanoemulsion: preparation, optimization, in vitro and in vivo evaluation | |
| Joseph et al. | Micelle/hydrogel composite as a “Natural Self-Emulsifying Reversible Hybrid Hydrogel (N’SERH)” enhances the oral bioavailability of free (unconjugated) resveratrol | |
| CN101301455A (en) | Chinese medicine compound turmeric rhizome solid dispersion for treating hyperlipemia | |
| US20210353555A1 (en) | Nanoparticles based on optionally alkoxylated poly(alkyl cyanoacrylates) having a defined degree of polymerization | |
| CN106581647A (en) | PH response insulin slow release nanoparticle, and preparation method and application thereof | |
| CN108524472B (en) | Nanosphere containing orlistat is preparing the purposes in anti-hepatic-B virus medicine | |
| Meng et al. | Characterization and pharmacokinetics of coenzyme Q10 nanoparticles prepared by a rapid expansion of supercritical solution process | |
| El-Zahaby et al. | Development of a novel solid self-nano-emulsifying osmotically controlled system of a centrally acting drug: preparation and in-vitro evaluation | |
| WO2021097652A1 (en) | Orlistat nanospheres and use thereof in preparation of medicines for treatment of obesity |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |