CN103845278B

CN103845278B - A kind of phylloxanthin eye nano-emulsion-thermosensitive in situ gel and preparation method thereof

Info

Publication number: CN103845278B
Application number: CN201210499309.0A
Authority: CN
Inventors: 韩静
Original assignee: Shenyang Pharmaceutical University
Current assignee: Shenyang Pharmaceutical University
Priority date: 2012-11-30
Filing date: 2012-11-30
Publication date: 2016-06-22
Anticipated expiration: 2032-11-30
Also published as: CN103845278A

Abstract

The present invention provides a kind of phylloxanthin eye nano-emulsion-thermosensitive in situ gel and preparation method thereof, when high speed shear and high pressure homogenize, phylloxanthin is made lutein nanometer breast with emulsifying agent, vegetable oil and appropriate water for injection first with wet gum method, then suitable thermosensitive hydrogel substrate is selected, add appropriate thickening agent, osmotic pressure regulator, antibacterial, antioxidant and stabilizer, prepare into one and present liquid condition in vitro, the ophthalmic preparation of umklappen gel state after instillation ophthalmic。Lutein nanometer breast mean diameter prepared by the present invention is for lower than 200nm, the corneal permeability of phylloxanthin and the adhesiveness of gel and slow corrosion can be improved well, increase drug loading, reduce drug release rate, extend drug treating time, improve local biologic availability。

Description

A kind of phylloxanthin eye nano-emulsion-thermosensitive in situ gel and preparation method thereof

Technical field

The invention belongs to field of medicine preparations, the preparation method relating to a kind of phylloxanthin eye nano-emulsion-thermosensitive in situ gel。

Background technology

Senile degeneration of macula is that one can cause central vision to decline, and seriously can cause blind oculopathy, and this disease is still obstinate disease at present。And the macula lutea in retina is had important protective effect by phylloxanthin, it is possible to effectively prevent and treat visual deterioration that senile eyeball retina macular degeneration causes and blind。And phylloxanthin is as antioxidant well and photoprotection, to preventing and treating the oculopathy such as cataract, all there is curative effect well。

Although the phylloxanthin product kind of listing is numerous at present, but it is oral formulations, ophthalmic preparation is considerably less, and traditional ophthalmic solution makes the time that medicine is detained before cornea very short because of the motion of eyeball and the effect of nose tear system, generally the eye drop of about 80-90% runs off instilling in eyes one minute after。The simultaneously barrier action of cornea, medicine also restrained across corneal osmosis, eye bioavailability is very low。For overcoming these not enough, the present invention adopts situ-gel technology。Situ-gel system is the fluid of low-viscosity in vitro, such as temperature, pH value etc. under physiological condition, self will produce the phase in version of so-gel, thus extending the precorneal residence time of medicine, dosage is accurate, the advantages such as reproducibility is good。

But phylloxanthin is a kind of water-insoluble material, its water-insoluble limits its application in ophthalmic preparation, phylloxanthin is first made lutein nanometer breast by the present invention, it is possible to increase the water solublity of phylloxanthin well, and Emulsion is made nanoscale can be greatly increased the corneal permeability of phylloxanthin；Prepare into situ-gel again, the plurality of advantages such as have again that bioadhesive is good and histocompatibility, bioavailability be high。

Summary of the invention

It is an object of the invention to solve the problem that phylloxanthin dissolubility is low, another purpose is in that add the corneal permeability of phylloxanthin and extend the medicine holdup time at eye, improves medicine and obtains local biologic availability。

The present invention is achieved through the following technical solutions:

The mass percent of its prescription is: phylloxanthin 0.01%-2%, emulsifying agent 0.1%-5%, vegetable oil 0.5%-10%, temperature sensitive hydrogel base starting material 2%-30%, thickening agent 0.05%-1%, osmotic pressure regulator 0%-5%, antibacterial 0.01%-5%, antioxidant 0.01%-5%, stabilizer 0%-5%, injection deionized water surplus。Wherein emulsifying agent optimum quality percentage ratio is 1%-4%, and vegetable oil optimum quality percentage ratio is 2%-8%, temperature sensitive hydrogel base starting material optimum quality percentage ratio 10%-25%。

Heretofore described phylloxanthin, for being raw material by Flos Tagetis Erectae, through organic solvent extraction, saponification, is refined and is obtained；Detecting carotenoid content more than 85% by GB26405-2011, lutein content is more than 75%。

Heretofore described emulsifying agent is that arabic gum, tragakanta, gelatin, fatty acid Pyrusussuriensis be smooth, a kind of in Polysorbate or their mixture；Wherein fatty acid Pyrusussuriensis smooth (Span class) is a kind of in Span20, Span40, Span60, Span80 or their mixture；Polysorbate (Tween class) is a kind of in Tween20, Tween40, Tween60, Tween80 or their mixture。

Heretofore described vegetable oil is a kind of in soybean oil, Oleum Ricini, olive oil, Oleum Arachidis hypogaeae semen or their mixture。

Heretofore described temperature sensitive hydrogel base starting material, for the one in chitosan, phosphoglycerol, xylan, poly-N-isopropyl acrylamide (PNIPAAm), poloxamer188, PLURONICS F87, Polyethylene Glycol section thing (PLGA-PEG-PLGA) altogether or their mixture。

Heretofore described thickening agent is one or more mixture in carboxymethyl cellulose, polycarbophil, methylcellulose, carbomer, PVPK30, PEG400。

Heretofore described osmotic pressure regulator is one or more mixture in mannitol, glucose, NaCl。

Heretofore described antibacterial is one or more mixture in benzalkonium chloride, benzalkonium bromide, methyl hydroxybenzoate, nipagin A, nipasol, chlorobutanol。

Heretofore described antioxidant is one or more mixture in sodium sulfite, sodium sulfite, sodium pyrosulfite, sodium thiosulfate, citric acid, vitamin C, vitamin E。

Heretofore described stabilizer is one or more mixture in polyvinyl alcohol, sodium lauryl sulphate, dodecylbenzene sodium sulfonate。

The present invention is prepared via a method which: is dissolved in vegetable oil by phylloxanthin powder under heating condition, makes oil phase；Emulsifying agent is joined in appropriate water for injection, makes aqueous phase；Being added by oil phase in aqueous phase, high speed shear emulsifying obtains colostrum, and then uses high pressure homogenizer homogenizing, makes nano-emulsion；Thickening agent, osmotic pressure regulator, antibacterial, antioxidant, stabilizer are joined in a certain amount of water for injection, adds temperature sensitive hydrogel substrate under ice bath, make gel solution；The nano-emulsion made is joined in gel solution, mix homogeneously, make phylloxanthin eye nano-emulsion-thermosensitive in situ gel。

The advantage that the phylloxanthin eye nano-emulsion-thermosensitive in situ gel of the present invention combines nano-emulsion and situ-gel。Compared with ordinary eye drops and ordinary gel agent, the invention have the advantages that (1) nano-emulsion particle diameter is little, mean diameter 163nm, not only can improve the dissolubility of phylloxanthin, stability, corneal permeability, it is also possible to delay release medicine, extend action time, be conducive to improving bioavailability;(2) in-situ gel had both overcome and the deficiency of ordinary eye drops and ordinary gel agent, the advantage maintaining again the two;(3) emulsifying agent used by, macromolecular material etc. to eye almost without zest and side effect。

This eye drop is storage-stable under 4 DEG C of conditions, uniform content, it is not necessary to secondary is prepared, thus reducing opportunities for contamination。Extend the preparation slow releasing function that simultaneously works as in the eye holdup time by gelation, improve the compliance of medicine local biologic availability and patient, it is possible to be applied to clinic, have broad application prospects。

Accompanying drawing explanation

Fig. 1 is that the phylloxanthin situ-gel prepared under embodiment 1 condition measures particle size distribution by particle size determination instrument。

Detailed description of the invention

Below in conjunction with specific embodiment, technical scheme is further described。

Embodiment 1:

Prescription 1

Phylloxanthin (lutein content 78%) 0.5g

Tween8025g

Oleum Ricini 50g

Poloxamer188 230g

PLURONICS F87 20g

Methylcellulose 10g

Mannitol 9g

Benzalkonium bromide 0.1g

Vitamin E 5g

Polyvinyl alcohol 1g

Water for injection adds to 1000ml

Detailed process is:

The first step, is dissolved in Oleum Ricini by recipe quantity phylloxanthin powder under 60 DEG C of conditions, makes oil phase；Recipe quantity Tween80 is joined in 10ml water for injection, makes aqueous phase；Being added by oil phase in aqueous phase, high speed shear 10000r min-1 obtains colostrum, then with high pressure homogenizer 100MPa homogenizing 5 times, makes nano-emulsion；

Second step, joins in 500ml water for injection by recipe quantity methylcellulose, mannitol, benzalkonium bromide, vitamin E, polyvinyl alcohol, adds recipe quantity poloxamer188 and PLURONICS F87, make gel solution under ice bath；

3rd step, joins in gel solution by the nano-emulsion made, mix homogeneously, injects water to 1000ml and makes phylloxanthin eye nano-emulsion-thermosensitive in situ gel, puts into 4 DEG C of cold preservations of refrigerator。

The mean diameter recording phylloxanthin situ-gel is 163nm。

Embodiment 2:

Prescription 2

Phylloxanthin (lutein content 82%) 0.1g

Arabic gum 10g

Oleum Ricini 50g

Chitosan 300g

Methylcellulose 10g

Mannitol 9g

Benzalkonium bromide 0.1g

Vitamin E 5g

Polyvinyl alcohol 1g

Water for injection adds to 1000ml

Concrete operation step and formula are with embodiment 1, and the mean diameter recording phylloxanthin situ-gel is 198nm。

Embodiment 3:

Prescription 3

Phylloxanthin (lutein content 80%) 20g

Tween2050g

Soybean oil 100g

Polyethylene Glycol is section thing (PLGA-PEG-PLGA) 250g altogether

Methylcellulose 10g

Mannitol 9g

Benzalkonium chloride 0.1g

Vitamin E 5g

Polyvinyl alcohol 1g

Water for injection adds to 1000ml

Concrete operation step and formula are with embodiment 1, and the mean diameter recording phylloxanthin situ-gel is 190nm。

Embodiment 4:

Prescription 4

Phylloxanthin (lutein content 90%) 0.5g

Tragakanta 1g

Oleum Arachidis hypogaeae semen 10g

Poloxamer188 18g

Chitosan 2g

Methylcellulose 10g

NaCl9g

Benzalkonium bromide 0.1g

Vitamin E 0.1g

Polyvinyl alcohol 1g

Water for injection adds to 1000ml

Concrete operation step and formula are with embodiment 1, and the mean diameter recording phylloxanthin situ-gel is 202nm。

Embodiment 5:

Prescription 5

Phylloxanthin (lutein content 85%) 1g

Span205g

Olive oil 5g

Monoglyceride 230

PLURONICS F87 20g

Carboxymethyl cellulose 0.5g

Mannitol 9g

Chlorobutanol 0.1g

Sodium sulfite 50g

Dodecyl sodium sulfate 1g

Water for injection adds to 1000ml

Concrete operation step and formula are with embodiment 1, and the mean diameter recording phylloxanthin situ-gel is 178nm。

Embodiment 6:

Prescription 6

Phylloxanthin (lutein content 88%) 10g

Span8025g

Oleum Ricini 50g

Poloxamer188 230g

Carbomer 10g

Glucose 50g

Methyl hydroxybenzoate 50g

Citric acid 50g

Polyvinyl alcohol 1g

Water for injection adds to 1000ml

Concrete operation step and formula are with embodiment 1, and the mean diameter recording phylloxanthin situ-gel is 176nm。

Embodiment 7:

Prescription 7

Phylloxanthin (lutein content 76%) 5g

Tween6025g

Oleum Ricini 50g

Xylan 230g

PEG40010g

NaCl9g

Benzalkonium bromide 0.1g

Vitamin C 5g

Polyvinyl alcohol 50g

Water for injection adds to 1000ml

Embodiment 8:

Prescription 8

Phylloxanthin (lutein content 91%) 10g

Gelatin 25g

Oleum Arachidis hypogaeae semen 50g

PNIPAAm150g

PVPK305g

NaCl25g

Benzalkonium bromide 25g

Vitamin E2 5g

Sodium lauryl sulphate 25g

Water for injection adds to 1000ml

Concrete operation step and formula are with embodiment 1, and the mean diameter recording phylloxanthin situ-gel is 200nm。

Embodiment 9:

Prescription 9

Phylloxanthin (lutein content 86%) 10g

Tween4025g

Soybean oil 50g

Poloxamer188 120g

PLGA-PEG-PLGA30g

PEG4005g

Benzalkonium bromide 0.1g

Vitamin E 5g

Polyvinyl alcohol 1g

Water for injection adds to 1000ml

Concrete operation step and formula are with embodiment 1, and the mean diameter recording phylloxanthin situ-gel is 182nm。

Embodiment 10:

Prescription 10

Phylloxanthin (lutein content 83%) 0.5g

Span4025g

Oleum Ricini 50g

Chitosan 100g

PLURONICS F87 20g

PVPK3010g

Mannitol 9g

Benzalkonium bromide 0.1g

Sodium thiosulfate 5g

Water for injection adds to 1000ml

Concrete operation step and formula are with embodiment 1, and the mean diameter recording phylloxanthin situ-gel is 188nm

Embodiment 11:

Prescription 11

Phylloxanthin (lutein content 89%) 0.5g

Span6025g

Oleum Ricini 50g

PLGA-PEG-PLGA230g

Phosphoglycerol 20g

Polycarbophil 0.5g

Mannitol 9g

Benzalkonium bromide 0.1g

Vitamin E 5g

Polyvinyl alcohol 1g

Water for injection adds to 1000ml

Concrete operation step and formula are with embodiment 1, and the mean diameter recording phylloxanthin situ-gel is 199nm。

The grain size distribution of phylloxanthin situ-gel: the phylloxanthin situ-gel prepared under Example 1 condition measures particle size distribution by particle size determination instrument, and result is shown in Fig. 1, and mean diameter is 163nm。

After phylloxanthin eye nano-emulsion-thermosensitive in situ gel administration, rabbit eye irritation is investigated: the phylloxanthin situ-gel prepared under Example 1 condition is investigated。

Administering mode:

In first 24 hours of on-test, the eyes of every animal are checked (including using fluorescein sodium inspection)。The animal having eye irritation symptom, corneal defect and conjunctival damage cannot be used for experiment。Take 3 rabbit numberings。Instilling O.1mL (1) phylloxanthin eye nano-emulsion-thermosensitive in situ gel in every rabbit conjunctiva of left eye capsule during test, right eye instills normal saline and compares。After administration, the eyelid about 10 seconds of gently sleeping。Only it is administered once。Instill 2% fluorescein sodium slit lamp examination and be administered the eye local excitation response situation of latter 0,6,24,48,72 hour to 7 days。

Ocular irritation is evaluated:

Ocular irritation evaluation methodology adopts perDraizetechnique, zest is divided into four grades (nothing, slight, moderate, severe) corresponding 0,1,2,3 points respectively, then the irritant reaction score value of the cornea of every animal, iris and conjunctiva is added, obtain the Ocular irritation reaction total mark of an animal subject, it is exactly this animal subject last score value to Eye irritation the irritant reaction integration summation of every animal subject divided by number of animals, evaluates animal subject Eye irritation degree by table 1。

Table 1 Ocular irritation evaluation criterion

To rabbit eye irritation result after table 2 phylloxanthin eye nano-emulsion-thermosensitive in situ gel administration

Result shows, after administration in 0h to the 7d time, eye irritation score value is 0, it was shown that phylloxanthin eye nano-emulsion-thermosensitive in situ gel is to rabbit eye irritation。

Phylloxanthin eye nano-emulsion-thermosensitive in situ gel rheology is investigated:

The each embodiment rheology of table 3 investigates (employing rotating cylinder viscometer)

Being shown by above-mentioned test result, the gel preparation environment viscosity in vitro that embodiment 1-11 prepares is all relatively low, and after mixing with artificial tears by 40:7 and be heated to 34 DEG C, its viscosity all dramatically increases。Thus, represent that said preparation can realize the effect of in-situ gelling。It is the relatively low liquid of viscosity in vitro, gelling occurring after eye drip, forming the clear gel that viscosity is bigger, to adhere to anterior corneal surface, thus extending the precorneal residence time of medicine, it is achieved better therapeutic effect。

Claims

1. phylloxanthin eye nano-emulsion-thermosensitive in situ gel, it is characterized in that: its prescription percentage by weight is: phylloxanthin 0.01%-2%, emulsifying agent 0.1%-5%, vegetable oil 0.5%-10%, temperature sensitive hydrogel base starting material 2%-30%, thickening agent 0.05%-1%, osmotic pressure regulator 0%-5%, antibacterial 0.01%-5%, antioxidant 0.01%-5%, stabilizer 0%-5%, injection deionized water surplus, described emulsifying agent is that arabic gum, tragakanta, gelatin, fatty acid Pyrusussuriensis be smooth, a kind of in Polysorbate or their mixture；Fatty acid Pyrusussuriensis is smooth for a kind of in Span20, Span40, Span60, Span80 or their mixture；Polysorbate is a kind of in Tween20, Tween40, Tween60, Tween80 or their mixture；Vegetable oil is a kind of in soybean oil, Oleum Ricini, olive oil, Oleum Arachidis hypogaeae semen or their mixture；Gel-type vehicle raw material is the one in chitosan, phosphoglycerol, xylan, poly-N-isopropyl acrylamide, Polyethylene Glycol section thing, poloxamer188, PLURONICS F87 altogether or their mixture；Thickening agent is one or more mixture in carboxymethyl cellulose, polycarbophil, methylcellulose, carbomer, PVPK30, PEG400, and is prepared via a method which: is dissolved in vegetable oil by phylloxanthin powder under heating condition, makes oil phase；Emulsifying agent is joined in appropriate injection deionized water, make aqueous phase；Being added by oil phase in aqueous phase, high speed shear emulsifying obtains colostrum, and then uses high pressure homogenizer homogenizing, makes nano-emulsion；Thickening agent, osmotic pressure regulator, antibacterial, antioxidant, stabilizer are joined in 500ml injection deionized water, adds temperature sensitive hydrogel base starting material under ice bath, make gel solution；The nano-emulsion made is joined in gel solution, mix homogeneously, make phylloxanthin eye nano-emulsion-thermosensitive in situ gel。

2. phylloxanthin eye nano-emulsion-thermosensitive in situ gel according to claim 1, it is characterised in that: in phylloxanthin, carotenoid content is more than 85%, and lutein content is more than 75%。

3. phylloxanthin eye nano-emulsion-thermosensitive in situ gel according to claim 1, it is characterised in that: osmotic pressure regulator is one or more mixture in mannitol, glucose, NaCl；Antibacterial is one or more mixture in benzalkonium chloride, benzalkonium bromide, methyl hydroxybenzoate, nipagin A, nipasol, chlorobutanol；Antioxidant is one or more mixture in sodium sulfite, sodium sulfite, sodium pyrosulfite, sodium thiosulfate, citric acid, vitamin C, vitamin E；Stabilizer is one or more mixture in polyvinyl alcohol, sodium lauryl sulphate, dodecylbenzene sodium sulfonate。

4. phylloxanthin eye nano-emulsion-thermosensitive in situ gel according to claim 1, it is characterised in that: heating-up temperature is 40-80 DEG C；High speed shear speed is 2000-20000r min^-1；High pressure homogenizer pressure is 20MPa-150MPa；Homogenization cycles is 2-10 time。