CN105982879B - A kind of Lamotrigine polymer micelle administration system of epileptogenic focus targeting - Google Patents
A kind of Lamotrigine polymer micelle administration system of epileptogenic focus targeting Download PDFInfo
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Abstract
The invention belongs to field of biotechnology, are related to the administration nano-drug administration system of epileptic focus targeting, and in particular to load Lamotrigine polymer micelle and its preparation method and application of the one kind based on tryptophan derivative (tryptophan derivate, TD) modification.The present invention contains small molecule hydrophobicity antiepileptic Lamotrigine, preparation can be targeted to the nano-micelle drug delivery system of epileptogenic focus using that can position to the Derivatives Modified Pluronic polymers micella of the targeting head tryptophan of epileptogenic focus.What prepared targeting micelle administration system can effectively improve hydrophobic drug enters brain-capacity power, by significantly improving drug after non-invasive intravenously administrable in the accumulation of brain and entering brain efficiency.Drug delivery system of the invention can significantly improve the ability that drug enters intracerebral epileptic focus, suitable for promoting other accumulations of hydrophobicity antiepileptic in epileptic focus, help and improve antiepileptic to the curative effect of intractable epilepsy.
Description
Technical field
The invention belongs to field of biotechnology, be related to administration nano-drug administration system of a kind of epileptic focus targeting and preparation method thereof and
Using, and in particular to load Lamotrigine polymerization of the one kind based on tryptophan derivative (tryptophan derivate, TD) modification
Object micella and its preparation method and application.
Background technique
It is one group caused by the brain neuron paradoxical discharge repeatedly characterized by epilepsy outbreak prior art discloses epilepsy
Chronic brain diseases are the second common diseases that illness rate is only second to cerebral apoplexy in the nervous system disease.It is reported that the existing epilepsy in China
Patient about 9,000,000, annual new cases 450,000, wherein 25%~30% patient passes through a variety of existing antiepileptics
(AEDs) regular treatment not can be effectively controlled breaking-out still, be treatment difficult point clinically, and cause disabled and death important
Reason.At present clinical treatment intractable epilepsy rely primarily on a variety of mechanism of action it is different AEDs combination or operation excision cause
Epilepsy lesion, but the former effect is limited and the latter can only be applicable in particular patient and side effect is larger, thus the application of the two by
Obvious limitation.Numerous studies are shown in recent years, and epileptic focus part blood-brain barrier (BBB) is more based on P- glycoprotein (P-gp)
The overexpression of the outer chlG of kind drug is to lead to the drug resistant important mechanisms of epilepsy, and the latter can be different by a variety of mechanism of action
AED pumps out lesion part and then influences curative effect of medication.Therefore, how in the outer chlG of effective bypass or specificity inhibition part
While activity is to increase drug concentration in local brain tissue, the normal physical protection function of blood-brain barrier (BBB) is not destroyed, from
And achieve the purpose that the curative effect for improving AED without increasing its side effect, it is Present clinical work urgent need to resolve generally acknowledged in the industry
One important topic, and the hot and difficult issue problem in epilepsy research at present.
Recently as the extensive use of nanotechnology, a new possibility way also is provided for the treatment of intractable epilepsy
Diameter.Had been found that in the research of the past nano material contain can effectively improve drug after some Neurotherapeutic drugs enter brain
Ability is without destroying normal BBB barrier function.Using Biodegradable polymeric material as transmitting and conveying therapeutic agent
The unique advantage of its targeting and controlled release is shown into the carrier of brain tissue, in the diseases such as central nervous system infection
Through entering clinical application.Researches show that the main mechanisms of most of nanoparticles across BBB to have: what nano medicament carrying system generally had
Long cycle characteristics may make it that can have more chances to be adsorbed on blood vessel, locally generate the concentration ladder across barrier
Degree, enters brain by Passive diffusion；The nanoparticle of surface modification ligands specific can be mediated or quiet by receptor-mediated, transporter
Electro Sorb mediate transcytosis and enter brain；In addition, certain nano materials (such as pluronics material) act not only as carrying
Body delivers drug, and itself can inhibit the outer pump activity of the albumen such as the P-gp on BBB, to reduce the resistance that drug enters brain
And increase the concentration of drug in local brain tissue.However, since to enter brain volume still limited for drug that administration nano-drug administration system is improved,
And it is relatively difficult to resolve the certainly intracerebral drug problem that non-selectivity is distributed in entire brain tissue, at present for the Brain targeting for overcoming BBB
Nanoparticle is also in experiment in vitro and animal experiment stage mostly, and adjusts nano material as vehicle delivery AEDs using biological respinse
Administration nano-drug administration system not yet appear in the newspapers.
Tryptophan is a kind of intracorporal necessary amino acid of people, can enter intracerebral by macromolecular neutral amino acid transporter body
It is converted into neurotransmitter serotonin.Focal due to abnormal kynurenine metabolism pathway in epilepsy, a large amount of tryptophans enter insane
Epilepsy stove intracellular metabolite is quinolinic acid and kynurenin, and the intake of local brain tissue tryptophan is caused obviously to be increased.It is multinomial in recent years
Research is using positron computer layer scanning (Epilepsia 2011,52 (9): 1692-1698；Neurology 2013,81
(7): 674-680) and MRI scan (Epilepsia 2008,49 (8): 1419-1430) finds to derive with tryptophan
Object (tryptophan derivate, TD) can effectively be gathered in epileptic focus as contrast medium prepared by ligand, thus real
Existing epileptic focus positioning.But at present both at home and abroad there is not yet carrying the nanometer of antiepileptic using the derivative as targeting head base building
The report of drug delivery system.
Summary of the invention
The purpose of the present invention is cannot be efficiently entering intractable epilepsy lesion part for clinical existing AEDs at present
Deng defect, provide a kind of nano-micelle drug delivery system for lesion part, and in particular to a kind of amino acid derivativges (color ammonia
Acid derivative) modification the nano target drug delivery system with bioactivity regulating power, more particularly to the new epileptogenic focus of one kind
The Lamotrigine polymer micelle administration system of targeting.
The present invention utilize amphiphilic Pluronic polymers material bioactivity regulating power, especially its had
Adjusting using P-gp to pump active ability outside a variety of multiple medicine transporters of representative, spread out with the tryptophan of great clinical value
Biological (TD) increases administration nano-drug administration system and penetrates as targeting head base, the Pluronic polymers carrier micelle of modified high molecular
The ability of blood-brain barrier, especially accumulation of the raising antiepileptic in epileptic focus, realize the lesion of AEDs target to
Medicine.
Specifically, epilepsy targeting of the invention carries Lamotrigine polymer micelle administration system, which is characterized in that use
Tryptophan derivative is as targeting head base, with the amphipathic nature block polymer pluronic P123 with biological respinse regulating power
As basic carrier material, using the polymer micelle F127 with tryptophan derivative group as component, the two is in deionization
The stable nucleocapsid structure of about 20nm is self-assembled into water or HBS solution or PBS solution (pH=7.4), by hydrophobicity model drug
Object Lamotrigine (LTG) is contained into the micelle inner core formed, be made epilepsy targeting of the invention carry LTG polymer micelle to
Medicine system.
The epilepsy target polymer drug delivery system of the tryptophan derivative modification is by carrier is carrier block copolymer
P123 and the block copolymer F127 auto-polymerization of tryptophan derivative modification are formed, it is preferable that as basic carrier
The weight ratio of Pluronic P123 and the F127 block copolymer of the tryptophan derivative modification as component is 7~8:3.
In the present invention, the pluronics material and its polymer with tryptophan derivative functional group derive
Object be selected from polyoxyethylene 20- polyoxypropylene 69- polyoxyethylene 20 (molecular weight 5750, pluronic P123 series, abbreviation P123),
Polyoxyethylene 100- polyoxypropylene 65- polyoxyethylene 100 (molecular weight 12600, pluronic F127 series, abbreviation F127) and color
The F127 (abbreviation TD-F127) of threonine derivative functionalization.
Selected Pluronic P123 and F127 raw material is provided by BASF Corp. of Germany in the present invention.
In the present invention, the carrier material is selected from pluronic P123 and F127, and safety and biocompatibility are equal
It is good, and the two has similar hydrophobic section and lower critical micelle concentration, therefore, the mixed with polymers micella tool formed
There is better solubilizing effect, the solubility of LTG can be obviously increased, and can keep good steady during a large amount of hemodilutions
It is qualitative.
In the present invention, the Pluronic P123 micella has biological respinse regulatory function, can effectively inhibit
The activity of P- glycoprotein penetrates various physiological and pathological barriers (blood-brain barrier, tumour screen so as to enhance the drug of its package-contained
Barrier) ability.
In the present invention, the drug that the polymer micelle of formation can be contained is delivered to by being passively or actively targeting approach
Specific tissue, by showing that modification has a specific molecular of targeting in polymer micelle, such as amino acid, peptides, carbohydrate,
Steroids, monoclonal antibody, agglutinin and compound of low molecular weight etc., can obviously increase it in the accumulation of target position, described poly-
Polyoxyethylene molecule (PEO) molecule for closing object micellar surface is able to extend the circulation time of micella in vivo, improves drug targeting
The probalility of success of delivering.
In the present invention, the target polymer micellar carrier of the described tryptophan derivative modification be by pluronic P123 and
The F127 composition of tryptophan derivative modification.
In the present invention, the tryptophan derivative modification polymer micelle F127 is delivered as hydrophobic drug
The component of carrier.
In the present invention, the tryptophan derivative is passed through by L-Trp methyl ester hydrochloride and benzyloxycarbonyl group-Beta-alanine
De-protected benzyl chloroformate group is taken off after Covalent bonding together to be formed.
The L-Trp methyl ester hydrochloride and benzyloxycarbonyl group-Beta-alanine molecule molar ratio is 1:1.
The L-Trp methyl ester hydrochloride and benzyloxycarbonyl group-Beta-alanine is in hexafluorophosphoric acid benzotriazole -1- base-oxygen
It reacts under the catalysis of base tripyrrole alkyl (PyBOP) and N, N- diisopropylethylamine (DIEA).The intermediate product of reaction is adopted
(Fig. 1-B) is characterized with nucleus magnetic hydrogen spectrum figure.
The product that above-mentioned reaction generates sloughs benzyl chloroformate group in ammonium formate and palladium carbon and generates required targeting
Head tryptophan derivative (tryptophan derivative, TD).
In the present invention, the amphipathic nature block polymer material can be selected from pluronics material, PEGylated phospholipid,
PEGylated chitosan class, PEGylated polylactic acid-based and PEGylated polyaminoacid class.
In the present invention, the amphipathic nature block polymer of the tryptophan derivative functionalization is the block copolymerization respectively
The product of the tryptophan derivative functionalization of the water-wet side of object material such as PEO molecule or PEG molecule.
In the present invention, the hydrophobic drug is selected from hydrophobic anti-infectives, anti-tumor drug or antiepileptic
Object；Especially hydrophobicity model drug LTG.
In the present invention, the F127 polymer micelle of the tryptophan derivative modification is by tryptophan derivative and carbonyl two
The F127 covalent bond of imidazoles activation forms.
The hydroxy activated of the end Pluronic F127 is the Pluronic F127 material that will purify in right amount and suitable
When carbonyl dimidazoles (CDI) reaction generation in anhydrous acetonitrile in the case where nitrogen protection of equivalent.
Wherein the molecule molar ratio of F127 material and CDI are 1:10.
The reaction system carries out at room temperature, and after reaction, reaction solution is concentrated through rotary evaporation, with pre-cooling
Anhydrous ether precipitating three times to remove unreacted CDI, collect precipitating, be dried in vacuo up to terminal activating F127-CDI powder
End.
In the present invention, the synthesis of the F127 polymer carrier materials of the tryptophan derivative modification is spread out by tryptophan
Biology reacts generation with F127-CDI in tetrahydrofuran.
The molecule molar ratio of the tryptophan derivative and F127-CDI is 10:1.
The present invention using nuclear magnetic resonance spectroscopy (1HNMR) technology to the various intermediate products of preparation process and final product into
Row Structural Identification (as shown in Figure 3), the results show that occurring unexistent feature on three unmodified F127 at 7-8.5PPM
Property absorption peak, be F127-CDI polymer micelle surface C DI group characteristic absorption peak；When changing tryptophan derivative-into
F127, the characteristic absorption peak of CDI disappears in spectrogram, absorption peak specific to multiple tryptophan derivatives occurs, shows color ammonia
Acid derivative successfully connects F127, and verifying tryptophan derivative-F127 target polymer micella synthesizes successfully.
The present invention provides the preparation methods for carrying LTG targeted nano micella comprising: color ammonia is prepared using film hydration method
The PF/LTG medicament-carried nano micelle of acid derivative modification: derive by the cool weighing pluronic P123 of optimizing prescriptions and with tryptophan
The F127 derivative of object radical functino is set in a round bottom flask, and acetonitrile is added and makes it dissolve；Hydrophobicity model drug is dissolved
It is configured to the stock solution of 3mg/ml in methanol, draws LTG stock solution and is added in the acetonitrile solution of pluronics material, two is molten
Liquid be sufficiently mixed after under the conditions of 50 DEG C rotary evaporation formed drug-polymer micelle membrane, be dried in vacuum overnight, be added prescription
The PBS (pH=7.4) of amount, 50 DEG C of water bath 200rpm isothermal vibrations make solution equilibria in 1 hour.Cross the organic filter membrane of 0.22um i.e.
Obtain the administration nano-drug administration system of epileptogenic focus targeting.
Specifically, the weight ratio of pluronic P123 and the F127 of tryptophan derivative functionalization are 7~8:3.
The present invention characterizes epilepsy targeted nanometer drug delivery system using particle instrument and transmission electron microscope, as a result such as Fig. 6
Shown, epilepsy targeted nano delivery system is particle of uniform, the spherical, partial size between 20-30nm.
The targeting of epilepsy constructed by the present invention carries LTG polymer micelle administration system and passes through pilocarpine Epilepsy
Tail vein injection mode, which is investigated, targets efficiency in body, and each area distribution in 1 hour hindbrain is the results show that carry LTG target polymer
Distribution of micella (TD-PF/LTG) group in cortex, cerebellum, hippocampus is above free LTG group, wherein in epileptic focus hippocampus
Drug concentration increase it is most obvious, be 2.7 times of free LTG, also increase by 90% compared to common micella (PF/LTG) group.
The target polymer micelle administration system of Rhodamine 123 (Rho123), brain are carried by epileptic rat tail vein injection
The case where fluorescence microscope qualitative observation Rho123 intracerebral is distributed after tissue freezing section, the epilepsy that the Primary Study present invention constructs
Target polymer micelle administration system enters brain mechanism.Since Rho123 is the classical substrate of highly expressed P-gp on BBB,
Free Rho123 is less to penetrate BBB, especially in the highly expressed epileptic focus region P-gp；From qualitative results as can be seen that
Polymer micelle after tryptophan derivative is modified can be obviously promoted Rho123 in the distribution of intracerebral, polymerize with unmodified
Object micellar phase ratio, can significantly improve fluorescence probe in the aggregation of hippocampus lesion.
Detailed description of the invention
Fig. 1 is the synthesis path and nucleus magnetic hydrogen spectrum figure (solvent DMSO-d for targeting head tryptophan derivative6)。
Fig. 2 is the synthesis path of the F127 of tryptophan derivative functionalization.
Fig. 3 hydrogen nuclear magnetic resonance spectrogram (1H-NMR the polymer P luronic F127 of different conditions) is characterized,
Wherein A:Pluronic F127 polymer micelle, solvent CDCl3,
B: the F127 (F127-CDI) of carbonyl dimidazoles functionalization, solvent CDCl3,
C: tryptophan derivative functionalization obtains F127 (F127-TD), solvent DMSO-d6。
Fig. 4 is the qualitative and quantitative result that BCECs absorbs fluorescence probe Rho 123 at different conditions.
Fig. 5 is that targeted medicament carrying nano micella prepares schematic diagram.
Fig. 6 is the partial size and Morphological Characterization for carrying LTG targeted nano micella.
Fig. 7 is point that the target polymer micella of fluorescence probe Rho 123 is observed under laser co-focusing in Different brain region
Cloth situation.
Fig. 8 is the intracorporal tissue distribution patterns of epilepsy targeted medicament carrying nano micella, the content with drug in each region of intracerebral
Enter the evaluation index of brain as drug than the blood concentration that desynchronizes.
Specific embodiment
Embodiment 1
By L-Trp methyl ester hydrochloride, 1:1 is dissolved in N, N- dimethyl formyl in molar ratio with benzyloxycarbonyl group-Beta-alanine
In amine, hexafluorophosphoric acid benzotriazole -1- base-oxygroup tripyrrole alkyl (PyBOP, 1.1eq) and n,N-diisopropylethylamine is added
(DIEA, 2eq) is poured into water, is extracted with ethyl acetate after room temperature reaction to raw material fully reacting, organic phase saturation NaCl
It washes twice, it is dry with anhydrous sodium sulfate, it is spin-dried for rear pillar chromatography and is prepared into intermediate product.Intermediate product is dissolved in methanol, is added
Ammonium formate (3eq), palladium carbon (10%, mass ratio), after back flow reaction 2 hours, after palladium carbon is filtered out, organic phase be spin-dried for after to obtain the final product
It can be used as the tryptophan derivative of targeting head base, synthesis path is as shown in Fig. 1-A；Pass through1H-NMR map tests product
Card, wherein the nuclear magnetic spectrogram result of compound 2 is as shown if figure 1-b: 1H-NMR (300MHz, DMSO-d6) δ: 10.85 (1H, s),
8.38-8.35 (1H, d, J=9Hz), 7.49-7.46 (1H, d, J=9Hz), 7.35-7.29 (6H, m), 7.21-7.17 (1H, t,
), J=6Hz 7.13-7.13 (1H, d), 7.08-7.03 (1H, t, J=9Hz), 6.99-6.95 (1H, t, J=6Hz), 4.99
(2H, s), 4.52-4.45 (2H, q, J=7.8Hz), 3.55 (3H, s), 3.17-2.97 (4H, m), 2.36-2.20 (2H, m).
Embodiment 2
The carbonyl dimidazoles functionalization of F127 terminal hydroxyl:
The Pluronic F127 material of appropriate (12.6g, 1mmol) acetone purifying is dissolved in 15mL anhydrous acetonitrile, separately will
1.62g carbonyl dimidazoles (CDI, 10mmol) are dissolved in 15mL anhydrous acetonitrile, then under nitrogen protection by the acetonitrile solution of CDI
In being slowly dropped in 2h in above-mentioned solution.After being added dropwise, hair answers system continuation to react 4h at room temperature, and reaction terminates
Afterwards, reaction solution is concentrated through rotary evaporation, with the anhydrous ether precipitating of pre-cooling three times to remove unreacted CDI, collects precipitating, very
Sky is drying to obtain terminal activating F127-CDI white crystalline powder；Pass through1H-NMR map verifies product.
Embodiment 3
The synthesis of the functionalization F127 of tryptophan derivative modification:
The final product that embodiment 1 synthesizes and the product F127-CDI that embodiment 2 synthesizes are dissolved in tetrahydro with molar ratio 10:1
In furans, after reflux 5 hours, overnight, after fully reacting, tetrahydrofuran is spin-dried for for room temperature reaction, and addition ether stirring is precipitated solid
Body, filter tryptophan derivative functionalization F127 (F127-TD)；Pass through1H-NMR map verifies product；Synthesis
Path and nucleus magnetic hydrogen spectrum figure are as shown in the figure.
Embodiment 4
Pluronic P123 and the 90mg color of drug-carrying polymer micelle weighing quantity fixing 180mg is prepared using film hydration method
The Pluronic F127 that threonine derivative is modified or do not modified is placed in a beaker, and acetonitrile 5ml is added and sufficiently dissolves, weighs Rameau
Triazine is dissolved in the stock solution that 3mg/ml is prepared into methanol, takes 5ml to be added in above-mentioned polymer solution, two solution are sufficiently mixed
Rotary evaporation forms drug-polymer micelle membrane under the conditions of 50 DEG C afterwards, is dried in vacuum overnight, the PBS (pH of recipe quantity is added
=7.4), 50 DEG C of water bath 200rpm isothermal vibrations make solution equilibria in 1 hour.0.22 μm of organic filter membrane is crossed up to Rameau three is carried
The nano-micelle drug delivery system of piperazine.
Embodiment 5
The nano-micelle of tryptophan derivative is modified with using Malvern nano-ZS current potential/Particle Size Analyzer measurement
(TD-PF/LTG) average grain diameter is 28.69 ± 0.29nm, as a result as shown in the figure.
Embodiment 6
It takes micellar solution appropriate, is added dropwise on the copper mesh of covering carbon film, drying at room temperature 0.5 hour, is placed under transmission electron microscope
The formalness of nano-micelle is observed, as a result as shown in the figure.
Embodiment 7
The Brain Microvascular Endothelial (BCECs) of logarithmic growth phase is made slender with 0.25% trypsin digestion
Born of the same parents' suspension is resuspended in serum-free DMEM after centrifugation, in being copolymerized burnt ware (35mm × 20mm) adjust cell density to 2 ×
104, observation state under light microscopic after being placed in 37 DEG C, cultivating 24 hours in 5%CO2 incubator, the normal person of form be separately added into containing
The PBS solution or blank PBS solution 1ml-1.5ml of 50mM glutamic acid, exhaustion supernatant is molten after above-mentioned the same terms are incubated for half an hour
Liquid adds DMEM incubation after washing 1 time with blank DMEM.Rhodamine 123 (Rho123) final concentration is added after 24 hours
For the Rho123PBS solution of 5uM (1.9ug/ml) or the PBS solution of 0.05%PF/Rho123, cultivated under the conditions of 37 DEG C of 5%CO2
30min, after wash 3 times with pre-cooling PBS, then plus PBS (cover ware bottom) a little, be placed directly under laser confocal microscope and see
It examines.As a result as shown, the polymer Rho123 that nano-micelle contains can significantly improve the ability that Rho123 enters BCECs,
And still there is similar effect on the BCECs of glutamic acid stimulation.
Embodiment 8
Single cell suspension is made with 0.25% trypsin digestion in the BCECs of logarithmic growth phase, suspends again after centrifugation
In PBS, adjustment cell density to 50 × 104.Take 50 × 104BCECs through or without the stimulation of glutamic acid shown in embodiment 7
Be resuspended in without serum without the Rho123 raw medicine that Rho123 final concentration of 5uM in phenol red DMEM culture solution, is added or
0.05%PF/Rho123 is in 37 DEG C, 5%CO2Incubator under be incubated for 30min.By the cell 1500r/min centrifugation 5 after incubation
Minute, and after being washed twice with ice-cold PBS, flow cytometer is analyzed, excitation wavelength 485nm, is collected at 530nm
Rho123 fluorescence carries out histogram analysis using Cell Quest software, gating when analysis, to remove cell fragment and cell mass
Interference of the block to measurement.With average fluorescent strength (the mean fluorescence of flow cytomery Rho123 intracellular
Intensity, MFI), MFI value is higher, and intake cellular uptake ability is stronger, and the outer pumping action of P-gp is weaker, on the contrary then on the contrary, result
As shown, Pluronic P123/F127 mixed micelle can effectively inhibit BCECs under physiology or pathological state to show P-gp
Function, effectively increase the penetration capacity of fluorescence probe Rho123.
Embodiment 9
In order to which the brain that enters of the Brain targeting polymer micelle of Primary Study building configures PF/ by 40mol% film hydration method
Rho123.0.8mg Rho123 and 35mg PF (P123:F127w/w 2:1,23mg P123,12mg F127 or F127-TD) are molten
In 5ml methanol, it is protected from light 1 hour formation film of rotary evaporation, addition 37 DEG C of PBS pH 7.42ml isothermal vibration 1 hour obtains molten
Liquid, 0.22um filtration can obtain the PF/Rho123 general polymer micellar solution and epilepsy targeting TD- of 0.4mg Rho123/ml
PF/Rho123 micellar solution gives the Rho that Rho123 and micella contain by the amount of every rat 3.5mg Rho123/Kg
123 solution, cardiac perfusion takes full brain after 1 hour, and 4 DEG C, fix 48 hours in 4% paraformaldehyde solution, 4 DEG C, 15% sugarcane
It is dehydrated 6 hours in sugar juice, 4 DEG C, be dehydrated 24 hours in 30% sucrose solution, carries out frozen section, thickness after embedding, freezing
It is 20 μm, nuclear staining is carried out using fluorescent reagent Hochest 33342 (1 μ g/ml), it is micro- with Carl-Zeiss laser co-focusing
Sem observation rat cerebral cortex, cerebellum, hippocampus Rho123 fluorescence distribution situation, as shown in the figure.
Pluornic P123 and the F127 micella used in the present invention is purchased from BASF AG, Rhodamine 123 (Rho123) purchase
From Sigma company.
Embodiment 10
PF/LTG general polymer micellar solution is prepared as described in Example 4 and the targeting of TD-PF/LTG epileptic focus is poly-
Close object micellar solution, SD rat (48 hours rats after normal rat or status epilepticus) difference tail vein injection 10mg
The common LTG solution of LTG/kg carries LTG general polymer micella and epilepsy target polymer micellar solution, is surveyed with HPLC method
Content of the LTG in intracerebral frontal cortex, cerebellum and hippocampus when 1 hour fixed.
Claims (8)
1. a kind of Lamotrigine polymer micelle administration system of epileptogenic focus targeting, which is characterized in that use tryptophan derivative
As targeting head base, use amphipathic nature block polymer pluronic P123 as basic carrier material, with derivative with tryptophan
The polymer micelle F127 of object group is self-assembled into stabilization in deionized water or HBS solution or PBS solution as component, the two
Nucleocapsid structure, by hydrophobicity model drug Lamotrigine contain into formed micelle inner core in, be made epileptogenic focus targeting
Lamotrigine polymer micelle administration system；
Wherein, the tryptophan derivative is by L-Trp methyl ester hydrochloride and benzyloxycarbonyl group-Beta-alanine through covalent bond
After slough blocking group and form, the molecule molar ratio of the two is 1:1.
2. the Lamotrigine polymer micelle administration system of epileptogenic focus targeting according to claim 1, which is characterized in that institute
The tryptophan derivative modification the stated polymer micelle transmits carrier as hydrophobic drug.
3. the Lamotrigine polymer micelle administration system of epileptogenic focus targeting according to claim 1, which is characterized in that institute
The Pluoronic F127 polymer micelle and amphipathic nature block polymer that the polymer micelle stated is modified by tryptophan derivative
Pluronic P123 composition, the P123 block copolymer of F127 and unmodified targeting head base that wherein tryptophan derivative is modified
Mass ratio be 1:2.
4. the Lamotrigine polymer micelle administration system of epileptogenic focus targeting according to claim 1, which is characterized in that institute
The pluronics stated and its derivative with carbonyl dimidazoles functionalization group are selected from polyoxyethylene 20 (PEO20)-polyoxy
The three block of propylene 69 (PPO69)-polyoxyethylene 20 (PEO20) and polyoxyethylene 100- polyoxypropylene 65- polyoxyethylene 100 is total
Polymers.
5. the Lamotrigine polymer micelle administration system of epileptogenic focus targeting according to claim 1 or 2, feature exist
In the F127 polymer micelle of the tryptophan derivative modification is by tryptophan derivative and carbonyl dimidazoles (CDI) function
The F127 block copolymer of change is reacted in tetrahydrofuran with the ratio of molecule molar ratio about 10:1.
6. the Lamotrigine polymer micelle administration system of epileptogenic focus targeting according to claim 5, which is characterized in that institute
In the F127 polymer micelle for the CDI functionalization stated, by the hydrophilic block polyoxyethylene of amphipathic nature block polymer F127 micella
The terminal hydroxyl of molecule is changed into the amphipathic nature block polymer that the carbonyl dimidazoles functionalization is made in carbonyl.
7. the Lamotrigine polymer micelle administration system of epileptogenic focus targeting according to claim 1, which is characterized in that institute
The hydrophobic drug stated is further selected from hydrophobic anti-tumor drug or anti-infectives.
8. the preparation method of the Lamotrigine polymer micelle administration system of epileptogenic focus targeting according to claim 1,
Be characterized in that comprising step: weigh recipe quantity amphipathic nature block polymer P123 and tryptophan derivative functionalization it is embedding
Section copolymer F127 is set in a round bottom flask, and methanol is added and makes it dissolve, weigh hydrophobic drug be dissolved in methanol be made it is certain
Round-bottomed flask is added in the stock solution of concentration, the drug methanol stock solution for drawing recipe quantity, and rotary evaporation is formed after mixing
Even drug-polymer film, is dried in vacuum overnight, and it is small that phosphate-buffered salt PBS, pH=7.4,50 DEG C of oscillation bed oscillations 1 are added
When make solution equilibria, filter membrane filtration method removes non-encapsulated drug, and the poly- of the epileptogenic focus targeting of tryptophan derivative modification is made
Close object micelle administration system.
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