CN110433132A - The tetrandrine nasal formulations for treating posttraumatic stress disorder - Google Patents
The tetrandrine nasal formulations for treating posttraumatic stress disorder Download PDFInfo
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Abstract
The invention discloses a kind of tetrandrine preparations for treating posttraumatic stress disorder.Tetrandrine is prepared into the nasal formulations such as temperature-sensitive gel, cubic liquid crystal, and the effect for the treatment of posttraumatic stress disorder well is played by Brain targeting.
Description
Technical field
The invention belongs to field of pharmaceutical preparations, and in particular to a kind of tetrandrine preparation for treating posttraumatic stress disorder.
Background technique
Posttraumatic stress disorder (post-traumatic stress disorder, PTSD) refers to Individual Experience or witnesses different
After ordinary catastrophic event (earthquake, big flood, war, traffic accident etc.), body delay is caused to occur or continue for a long time existing
A kind of phrenoblabia.The clinical manifestation of PTSD mainly has traumatic symptom, avoidance and numb class symptom, the alertness of recapturing to increase
Symptom etc..Treatment for PTSD, presently preferred serotonin reuptake inhibitors (Selective Serotonin
Reuptake Inhibitor, SSRI), wherein Prozac, Paxil, Sertraline curative effect are preferable, but long-term administration institute band
The side effect come is also that cannot be neglected, and common are nausea, headache, insomnia, serious person can also cause blood and lymphatic systems
Discomfort, immune disorder, sex dysfunction etc..
Tetrandrine is a kind of double benzyl isoquinolines from extracting in the dry root of Chinese medicine Stephania tetrandra section plant Fourstamen Stephania Root
Quinoline Alkaloid, belongs to calcium channel blocker, is clinically mainly used for the treatment such as cardiovascular disease, rheumatism, has anti-inflammatory, anti-
The pharmacological actions such as tumour and immunological regulation.Tetrandrine structural formula is as follows.
In the present invention, temperature-sensitive gel be it is a kind of be administered at room temperature for solution and with liquid condition, with
Because temperature increases phase transition occurs for medicine position, and the preparation of gel is converted by solution.Cubic liquid crystal is by amphipathic in the present invention
The system that molecular melting is formed in polar solvent has many advantages that mobility is low and bioadhesive as pharmaceutical carrier
Greatly, skin or mucous membrane surface can be attached to for a long time, improve the absorption of drug.
The addition of bio-adhesive material in addition to lasting medicine, it is easy to use, the bioadhesions such as first pass effect can be avoided
Outside the characteristic of agent, it also has the characteristics that, and good dispersion, adhesion strength are strong, stability is good, medicine-feeding part local concentration is high.
Summary of the invention
The present invention provides a kind of nasal formulations composition, can effective antidepression and anti-for treating posttraumatic stress disorder
Anxiety symptom, more existing dosage form is curative for effect, easy to use, and preparation process is simple.
The present invention provides a kind of nasal formulations composition, and it includes drug solutions, temperature-sensitive gel.Said medicine is molten
Liquid, raw material contain 0.1~10 part of tetrandrine, 10~30 parts of hydrochloric acid (0.1mol/L) in parts by weight.
The temperature-sensitive gel, raw material contain 0.1~10 part of PLURONICS F87,1~10 part of pool Lip river in parts by weight
407,15~35 parts of water of husky nurse.
Here is advanced optimized to foregoing invention, in above-mentioned temperature-sensitive gel, preferably PLURONICS F87 with
Poloxamer188 weight ratio is 1: 5~7.
The preparation method of nasal formulations composition provided by the invention, comprising the following steps: (1) by nasal formulations composition
In include poloxamer188 and PLURONICS F87 be added ultrapure water be sufficiently swollen at 4 DEG C, continue to add water to full dose, mix
Afterwards up to temperature-sensitive gel；(2) temperature sensitivity is added in the drug solution or suspension that include in nasal formulations composition
Property gel in mix to get nasal gel preparation.
The present invention provides a kind of tetrandrine nasal formulations composition, raw material contains 0.1~10 part of powder in parts by weight
Menispermine, 30~90 parts of cubic liquid crystals.
The preparation method of nasal formulations composition provided by the invention, comprising the following steps: (1) will be equipped with single oleic
The centrifuge tube of ester, which is put into, to be heated to glyceryl monooleate in 45 DEG C of thermostat water baths and melts completely, and ethyl alcohol is added, and vortex keeps it mixed
Close uniformly, as oily phase: (2) are added drug and mix；(3) absorption is preheated to 45 DEG C of deionized water, adds water droplet in vortex
Into oily phase, place 7 days at room temperature after mixing evenly.
The preparation method of nasal formulations composition provided by the invention, comprising the following steps: (1) add in glyceryl monooleate
Enter ethyl alcohol, the container heating water bath of the two will be filled, until glyceryl monooleate Quan Rongzhi clear transparent solutions；(2) by Fourstamen Stephania Root
Alkali and water mix, same to put 45 DEG C of heating；(3) centrifuge tube is held on eddy mixer, and it is mixed to draw tetrandrine with 1ml syringe
Suspension is added dropwise rapidly, until centrifugation liquid in pipe gradual change stiff, continues to stir, continues that surplus solution is added dropwise.
Tetrandrine has the pharmacological actions such as anti-inflammatory, antitumor and immunological regulation.Temperature-sensitive gel is easy to wrap up medicine
Object, bioadhesive is strong, can extend residence time of the drug in nasal cavity, and drug effect and easy to use can be improved, and toxic side effect is small,
There is good curative effect to posttraumatic stress disorder.
Detailed description of the invention
32 DEG C of < of temperature-sensitive gel appearance .A:T of Fig. 1 tetrandrine, 32 DEG C of B:T >
Fig. 2 tetrandrine temperature-sensitive gel rheological property
In Fig. 3 temperature-sensitive gel nasal cavity the residence time administration after 0h (A), 0.5h (B), 1h (C), 2h (D), 4h (E)
Influence .A: physiological saline group of Fig. 4 difference preparation to toad palate fibre swing；B: deoxysodium cholate group；C: temperature
Spend sensitive gels group；D: tetrandrine temperature-sensitive gel group (n=4,).
The test of Fig. 5 Elevated plus-maze proves PTSD model foundation success .A: normal mouse path；B: model mice road
Diameter；C: administration advances into open arms percentage；D: administration advances into open arms incubation period；E: residence time in open arms before being administered (n=6,) compared with normal group,*P < 0.05,**P < 0.01
The pharmacodynamics .A of Fig. 6 Elevated plus-maze test evaluation tetrandrine temperature-sensitive gel treatment PTSD: normal
Mouse path；B: model mice path；C: tetrandrine temperature-sensitive gel treatment group mouse path；D: enter after administration and open
Arm percentage；E: enter open arms incubation period after administration；F: residence time in open arms after administration (n=6,) and normal group ratio
Compared with,*P < 0.05,**P < 0.01,***P < 0.001；Compared with model group,#P < 0.05,##P < 0.01,###P < 0.001
Fig. 7 tetrandrine temperature-sensitive gel mouse deadlock is lived the time influence (n=3,) and normal group ratio
Compared with,*P < 0.05,**P < 0.01；Compared with model group,#P < 0.05,##P < 0.01
Hippocampus, prefrontal cortex, amygdaloid nucleus H.E. pathological section (× 200) in Fig. 8 difference group mouse
Hippocampus, prefrontal cortex, amygdaloid nucleus c-fos protein immunization group (× 200) in Fig. 9 difference group mouse
Illustrate technical solution of the present invention below with reference to specific embodiment, but protection of the invention is without being limited thereto.
The present invention provides a kind of tetrandrine nasal formulations composition, raw material contains 0.1~10 part of powder in parts by weight
Menispermine, 10~30 parts of hydrochloric acid (0.1mol/L), 50~70 parts of temperature-sensitive gels.
In a preferred embodiment, raw material contains 0.1~5 part of tetrandrine, 15~20 portions of salt in parts by weight
Acid (0.1mol/L), 40~50 parts of temperature-sensitive gels.
The temperature-sensitive gel includes following combination in parts by weight, 0.1~10 part of PLURONICS F87, and 1~10
Part poloxamer188,15~35 parts of water.
The present invention provides a kind of tetrandrine nasal formulations composition, raw material contains 0.1~10 part of powder in parts by weight
Menispermine, 30~90 parts of cubic liquid crystals.
Specific embodiment
Embodiment one: tetrandrine nose mixed suspension preparation
It takes 2g PLURONICS F87 and 12g poloxamer188 that suitable quantity of water is added sufficiently to be swollen at 4 DEG C, continues to add water to
50ml obtains temperature-sensitive gel after mixing；1g tetrandrine is taken, 6ml 1,2-PD and 8ml ethyl alcohol is added, mixing is equal
It is even；Above-mentioned tetrandrine suspension is added into the above-mentioned temperature-sensitive gel of 36ml, tetrandrine nasal formulations are obtained after mixing.
Embodiment two: tetrandrine nose cubic liquid crystal
1. prescription
2. preparation process
Water-bath keeps 45 DEG C of constant temperature, weighs 3.2g glyceryl monooleate and is put in 50ml centrifuge tube, and ethyl alcohol 0.3g is added,
Centrifuge tube is placed on heating water bath in the beaker in water-bath, until glyceryl monooleate Quan Rongzhi clear is molten in centrifuge tube
Liquid.0.75g tetrandrine, 1.5g water are put in beaker and are mixed, with 45 DEG C of the bath heating that discharge water.Hand-held centrifuge tube is mixed in whirlpool
In clutch, tetrandrine suspension is drawn with 1ml syringe, is added dropwise rapidly, until centrifugation liquid in pipe gradual change stiff, continues
Stirring continues that surplus solution is added dropwise.
Embodiment three: tetrandrine nose cubic liquid crystal
Precision weighs 3.2g glyceryl monooleate and is placed in centrifuge tube, and centrifuge tube is put into 45 DEG C of thermostat water baths and is heated
Melt completely to glyceryl monooleate, 0.3g ethyl alcohol is added, vortex is uniformly mixed it, as oily phase.0.1g Fourstamen Stephania Root is added
Alkali is vortexed again for mixing, and draws the deionized water that 1.4g is preheated to 45 DEG C with 1ml syringe, water droplet is added to oil in vortex
Xiang Zhong, glass bar are placed 7 days at room temperature after mixing evenly.
Example IV: tetrandrine nose temperature-sensitive gel (tetrandrine temperature sensitive
Gels, TTG) preparation
(1) above-mentioned temperature-sensitive gel is prepared；(2) precision weighs 0.2g tetrandrine in 10mL centrifuge tube, is added
3mL hydrochloric acid (0.1mol/L) is uniformly mixed；(3) mixture in (2) is added in 7mL temperature-sensitive gel, vortex mixer
Being sufficiently mixed uniformly makes dissolution to get tetrandrine nose temperature-sensitive gel.
Experimental example one: the properties evaluations of tetrandrine nose temperature-sensitive gel
1. evaluation method
Gelation temperature and gelling time, which are measured, is placed in thermostat water bath for the test tube equipped with TTG, and test tube solution face is low always
In water-bath face, bath temperature is slowly increased to 40 DEG C by 20 DEG C, and heating rate is 1 DEG C/min, reverses test tube, observation liquid flow is emotionally
Condition, temperature when liquid does not flow are denoted as gelling temp, and liquid is changed into the curdled appearance time by solution state as gelling time.
Rheological property investigates the rheological property using the advanced rotational rheometer measurement TTG of Anton Paar, investigates in 10-
The variation of TTG loss modulus (G ') storage modulus (G ") and compound viscosity (Viscosity) in 40 DEG C of temperature ranges；Investigate 32
Under the conditions of DEG C, in 0.1-100rad/s shearing frequency range, TTG loss modulus (G ') storage modulus (G ") and compound viscosity
(Viscosity) variation.
The investigation of gel nasal-cavity residence time weighs 10mgCy7 and adds to (2mg/ in the above-mentioned blank temperature-sensitive gel of 5ml
Ml), while same concentrations Cy7 aqueous solution is prepared.ICR mouse is randomly divided into blank control group, Cy7 solution group and Cy7 temperature
Sensitive gels group, Cy7 solution group mouse give 20 μ L Cy7 aqueous solutions (2mg/mL), Cy7 temperature-sensitive gel group mouse
Equivalent Cy7 temperature-sensitive gel (2mg/mL) is given, blank control group is with no treatment.Select 745nm as excitation wave
It is long, respectively upon administration 0,1,2,4h carries out toy fluorescence imaging and observes.
Ciliary toxicity, which is investigated, is randomly divided into physiological saline group (negative control), deoxysodium cholate for 12 bufo gargarizans Cantors
Group (positive control), blank temperature-sensitive gel group and TTG group, every group 3.After toad is put to death, palate is exposed, ophthalmology is used
It cuts and carefully separates palate mucous membrane with tweezers, area is about 5mm × 5mm, normal saline flushing clot and tissue.By palate mucous membrane
Cilium is placed on glass slide up, and 100 μ L physiological saline, 1% deoxysodium cholate, blank temperature-sensitive gel are added dropwise respectively
And TTG, gently covered, in 200 times of optical microphotograph microscopic observation toad palate fibre swing situations, record is from giving
Medicine starts to cilium to stop swinging duration, i.e. cilium persistent movement time (Persistent vibration
Duration, PVD).With the PVD of each administration group toad divided by the PVD of physiological saline up to cilium persistent movement percentage
(Percentage of persistent vibration, PPV).PPV is higher, and expression drug is smaller to the toxicity of cilium, on the contrary
It is bigger.
2. result
TTG gelation temperature and at 4 DEG C of gelling time, TTG appearance is the sticky flowing shape liquid (Figure 1A) of white.At 37 DEG C,
Rapidly transform into not flow-gel state (Figure 1B).It is lower than 32 DEG C by the gelation temperature that test tube roll back method measures TTG, gelling time
For 1.32min.
Rheology shows that there is TTG temperature sensitivity storage modulus to represent semi-solid state, and loss modulus represents liquid
State.For temperature at 23 DEG C, storage modulus is equal to loss modulus, this temperature is the critical-temperature that liquid changes to semisolid.When
When temperature is lower than 23 DEG C, loss modulus is higher than storage modulus, and the Viscous Characteristics of liquid are dominant, illustrates temperature sensitivity at this time
Gel is the liquid of flowing；When temperature be higher than 23 DEG C when, storage modulus be greater than loss modulus, the elastic characteristic of solid account for it is leading,
Gel state at this time is semisolid.After temperature is increased to 35 DEG C, storage modulus and loss modulus tend towards stability, and store up
Energy modulus is greater than loss modulus, therefore under nasal cavity temperature, TTG is attached in nasal cavity with semi-solid state, increases in nasal cavity
Residence time.Temperature is 32 DEG C, and shearing frequency ω is in 0.1-100rads-1When in range, storage modulus is always more than loss
Modulus, TTG are constantly in semi-solid state, show under environment in nasal cavity, and the solid property of preparation is better than liquid property, gel always
Stable structure；As shearing frequency increases, gel viscosity becomes smaller, and there are shear shinning phenomenon (Fig. 2).
TTG nasal cavity increased retention extends at any time, Cy7 solution group and Cy7 temperature-sensitive gel group mouse nasal cavity
Middle fluorescence intensity is gradually reduced, and wherein the decline of Cy7 normal saline solution group is very fast, the decline of Cy7 temperature-sensitive gel group compared with
Slowly.Solution good fluidity is easily discharged by intranasal ciliary movement, is not detained in nasal cavity, flowed to quickly by nasal cavity after administration
In oral cavity.And temperature-sensitive gel has bioadhesive, 0.5 after administration, 1,2, fluorescence intensity is above Cy7 in 4h nasal cavity
Normal saline solution (Fig. 3).Illustrate at least stop 4h or more in nasal cavity using gel as pharmaceutical carrier, extends drug in nose
The intracavitary residence time reduces drug wastage caused by nose fibre swing, improves drug bioavailability.
TTG influences smaller physiological saline group toad palate ciliary structures to toad palate fibre swing and arranges
Neatly, high-visible, movement is active, regularly swings to same direction in billowing wheat formula, and hunting frequency is higher.When giving
After 1% deoxysodium cholate, mucous membrane surface arrangement is disorderly and unsystematic, and ciliary structures are seriously damaged, and stops swinging in a short time, portion
Divide and falls off, toxicity maximum (Fig. 4).Blank temperature-sensitive gel group and TTG group, ciliary structures are more visible complete, and movement is relatively living
Jump, does not occur obvious cell detachment phenomenon, shows blank temperature-sensitive gel and TTG to cilium almost without toxicity, safety
Property is higher, can be used for nasal-cavity administration.The PVD and PPV of each administration group put in order from high to low is followed successively by physiological saline group > sky
White temperature-sensitive gel group > TTG group > deoxysodium cholate group (table 1).Compared with physiological saline group, deoxysodium cholate group
PVD and PPV significantly reduces (P=0.000 < 0.001)；Compared with deoxysodium cholate group, blank temperature-sensitive gel and TTG
PVD, PPV of group significantly increase (P=0.000 < 0.001), illustrate that blank temperature-sensitive gel and TTG safety are higher.
The different prescriptions of table 1 to fibre swing influence (n=3,).
Note: compared with physiological saline group,*P < 0.05,**P < 0.01,***P < 0.001；Compared with deoxysodium cholate group,#
P < 0.05,##P < 0.01,###P < 0.001
Experimental example two: the pharmacodynamic evaluation of tetrandrine nasal formulations composition treatment posttraumatic stress disorder
1 materials and methods
1.1 experimental animals and grouping
Experimental animal health ICR mouse 18, SPF grades, weight is 20 ± 1g, ties up tonneau China experimental animal purchased from Beijing
Technology Co., Ltd., credit number: SCXK (capital) 2016-0006.Animal feeding environment: 21-25 DEG C, 12h illumination/12h is given
Dark, ad lib drinking-water.Healthy C57BL/6N mouse 9, SPF grades, weight is 20 ± 1g, purchased from the experiment of Beijing dimension tonneau China
Zoo technical Co., Ltd, credit number: SCXK (capital) 2016-0006.Animal feeding environment: 21-25 DEG C, give 12h illumination/
12h is dark, ad lib drinking-water.
1.2 dosage regimens and evaluation
The extension of mouse single stress (Single Prolonged Stress, SPS) all Animal adaptabilities of model foundation
After raising 3d, it is randomly divided into normal group, SPS model group, TTG treatment group, every group 6, totally 18.
The foundation of SPS model is divided into three steps:
(1) confine: mouse being placed in 50mL centrifuge tube (centrifuge tube is perforated to guarantee animal eupnea), is horizontally arranged and prohibits
Plug with molten metal 2h；
(2) forced swimming: the mouse after confinement is put into depth of water 11cm, forces trip in the 2L beaker of water temperature (25 ± 1) DEG C
Swim 20min；
(3) etherization: the mouse after forced swimming being put into after restoring 15min in rearging cage, will be small with anhydrous ether
Mouse is anaesthetized to the loss of consciousness, after place it in ventilation and wait for that it revives naturally, then normal raising.
Dosage regimen starts to be administered with 2d after the success of evaluation index model foundation, and TTG group single-nostril gives 20mg
kg-120 μ L of TTG, one time a day, continuous 7d, normal group and model group are not administered.
Elevated plus-maze mouse elevated plus-maze (Elevated plus maze, EPM) by long-armed, galianconism and in
Centre area is formed by connecting, long-armed a length of 700mm, width 80mm；The a length of 665mm of galianconism, width 55mm, bottom and surrounding are black
Color is integrally placed on liftable Stainless Steel bracket, and the distance for adjusting Elevated plus-maze pedestal to ground is 80cm.Model is built
2d after vertical is freely put down mouse head towards open arms direction by middle section, record mouse in 300s respectively enter open arms and
The number of closure arm and residence time, to verify whether SPS model is successfully established.EPM detection is carried out again after administration 7d, is evaluated
Whether TTG has anti-PTSD effect.Excrement is cleared up after every mouse experiment, with 75% alcohol wipe, last dry wipe
Next round experiment is carried out after drying, and avoids the influence of excreta and smell to experiment mice.
Mouse conditioned fear model foundation conditioned fear is tested all Animal adaptabilities in (inevitability foot shock) and is raised
After supporting 3d, it is randomly divided into normal group, conditioned fear model group, TTG treatment group, every group 3, totally 9.
Conditioned fear model foundation is that mouse is put into the stimulation put under house arrest and receive two stages in electric shock case:
(1) the spontaneous activity stage: mouse is put into put under house arrest electric shock case in move freely 300s, do not give any stimulation；
(2) stimulation period: (shock parameters: 0.8mA is spaced 10s to 15 intermittent, inevitability foot shocks, continues
10s, total 300s).
Above-mentioned steps continue two days, and normal group is placed in and puts under house arrest electric shock case, but does not give and shock by electricity, total 600s.Every mouse reality
Excrement is cleared up after testing, and puts under house arrest electric shock bottom portion and side with 75% ethanol, avoids excreta and smell to experiment
The influence of mouse.
Dosage regimen starts to be administered with 2d after the success of evaluation index model foundation, and TTG group single-nostril gives 20mg
kg-120 μ L of TTG, one time a day, continuous 7d, normal group and model group are not administered.
After mouse deadlock lives in behavioral value administration 7d, animal is placed in electric shock case and carries out Environmental re-appearing experiment, does not give and appoints
What stimulation (every animal is placed on similarly shocks by electricity in case with when modeling), using video analysis software detection mouse in 300s
Dead time.
H.E. pathology section examination conditioned fear model mice takes complete after administration 7d and stiff firmly behavioral value after anesthesia
Brain tissue, which is placed in 4% formaldehyde, to be saved fixation, dehydration, paraffin embedding, thinly slices, hematoxylin-eosin (H.E.) dyeing, and 200
The variation of times optical microphotograph microscopic observation cerebral morphology[23]。
Immunohistochemistry detects c-fos protein expression inevitability foot shock model mice in administration 7d and stiff firmly behavior
After detection, anesthesia Hou Qu whole brain tissue, which is placed in 4% formaldehyde, to be saved, and is cut hippocampus and prefrontal cortex cross section, is seen under 200 times of mirrors
Examine the expression of c-fos albumen.
Statistical method analyzes software using SPSS16.0, and data use one-way analysis of variance, P < 0.05 between multiple groups
Indicate significant difference, experimental data withIt indicates.
2 experimental results
2.1 Elevated plus-mazes verify model foundation success
From normal, two groups of model path analysis, compared with normal group, the time and distance that model group is detained in open arms are equal
It is obviously shortened, shows that single extends Stress model and is successfully established (Fig. 5 A, B).By tri- index analysis of OE, OL and OT, and just
Normal group is compared, and the OE (Fig. 5 C) of model group is obvious than normally organizing low (P=0.009 < 0.01), and OL (Fig. 5 D) is than normally organizing high (P
=0.033 < 0.05), OT (Fig. 5 E) shows the success of SPS model foundation than normally organizing short (P=0.004 < 0.01).
2.2 Elevated plus-mazes prove that tetrandrine temperature-sensitive gel has anti-PTSD sample effect
Compared with normal group, model group is obviously shortened in the time that open arms is detained with distance, shows that single extension stress mould
Type is successfully established.Compared with model group, TTG group is significantly increased in the time that open arms is detained with distance, shows that TTG has PTSD
Good improvement result (Fig. 6 A, B, C).Through statistical analysis, compared with normal group, model group OE is substantially reduced (P=0.000
< 0.001), OT reduce but it is unobvious (P=0.101 > 0.05)；OL increases (P=0.671 > 0.05) than normal group.Although OL
And OT index no difference of science of statistics, but still have anxiety phenomenon compared with normal group, further prove the success of SPS model foundation.With
Model group is compared, and OE, OT obviously increase (P=0.002 < 0.01, P=0.033 < 0.05), OL after TTG single-nostril administration 7d
It reduces (P=0.201 > 0.05) (Fig. 6 D, E, F).Although OL no difference of science of statistics, TTG group still has improvement compared with model group
Trend, show TTG have good anti-PTSD effect.
2.3 stiff firmly behaviors illustrate that TTG has the function of good anti-PTSD
Compared with normal group, the model group dead time obviously increases (P=0.004 < 0.01), and it is small to illustrate that Environmental re-appearing makes
Mouse generates fear memory, the success of conditioned fear model foundation；Compared with model group, the dead time of TTG group significantly reduces (P=
0.049 < 0.05), show that TTG has the function of good anti-PTSD (Fig. 7).
2.4 H.E. pathological sections illustrate that TTG can obviously reverse the lesion of hippocampus, prefrontal cortex and amygdaloid nucleus position
Hippocampus (area CA1, the area CA2, the area CA3, the area DG), prefrontal cortex are observed by inverted fluorescence microscope (× 200)
And almond nuclear structure, it is found that normal group hippocampus, the equal caryogram of prefrontal cortex and amygdaloid nucleus position are regular, marshalling.With it is normal
Group is compared, and the area model group CA1, prefrontal cortex and amygdaloid nucleus position caryogram are irregular, misaligned uneven, and is occurred
The phenomenon that karyopycnosis contaminates deeply, shows that inevitability foot shock causes the area CA1, prefrontal cortex and amygdaloid nucleus position that disease has occurred
Become.Compared with model group, TTG group karyopycnosis dye phenomenon deeply disappears, and Cell nuclei spreads are neat, illustrate that TTG can alleviate PTSD institute
Cause mouse pathological change.For the area hippocampus CA2, the area CA3 and the area DG, normal group, model group, TTG group and indifference, therefore just
Step judges PTSD diseased region mainly at the area CA1 of hippocampus, prefrontal cortex and amygdaloid nucleus position[24](Fig. 8).
2.5 ImmunohistochemistryResults Results illustrate that TTG can significantly reduce c-fos protein expression caused by foot shock
C-fos participates in nerve fibre to the anti-of noxious stimulation as the fast reaction gene in proto-oncogene family
It answers, shows the excitability of neuron.Inverted fluorescence microscope (× 200) observation hippocampus (area CA1, the area CA2, the area CA3, the area DG),
Prefrontal cortex and almond nuclear structure, blue are nucleus, and brown color is c-fos positive signal.After PTSD model foundation, hippocampus
The expression quantity in the area CA1, prefrontal cortex and amygdaloid nucleus position c-fos significantly increases, and the area hippocampus CA2, CA3, DG has no obvious
Difference.TTG after one week of dosing, can reduce the expression quantity of Hippocampal CA 1, prefrontal cortex and amygdaloid nucleus position c-fos, show
TTG has good improvement result to PTSD symptom caused by foot shock, and the diseased region of mouse is mainly in hippocampus CA1
Area, prefrontal cortex and amygdaloid nucleus position (Fig. 9), it is consistent with H.E. result.
3 conclusions
By Elevated plus-maze carry out Behavioral assessment, mouse PTSD stress after, open arms enters number ratio and opens
The arm residence time, it is higher than normally organizing that open arms enters incubation period obviously than normally organizing reduction, shows that animal is in anxiety state；TTG
Nasal-cavity administration after a week, carries out Elevated plus-maze evaluation again, and three indexs show that the mood of anxiety has greatly change, show
TTG can improve the mood of mouse anxiety, generate the effect of anti-PTSD.Deadlock lives behavioral value the results show that foot shock can
So that mouse is generated fear memory, extends the dead time, and TTG can then shorten dead time of the mouse in fear of confined spaces case, into
And generate the effect of anti-PTSD.H.E. and c-fos immunohistochemistry pathological section reflects the diseased region of PTSD mainly in hippocampus
The area CA1, prefrontal cortex and amygdaloid nucleus.In conclusion tetrandrine temperature-sensitive gel is as a kind of traditional Chinese medicine monomer brain
Targeting preparation, the nasal cavity residence time is long, highly-safe, can significantly improve PTSD symptom, generates the effect of anti-PTSD.
Claims (11)
1. a kind of for treating the nasal formulations composition of posttraumatic stress disorder, wherein comprising drug solution, drug suspension,
Temperature-sensitive gel.
2. a kind of for treating the nasal formulations composition of posttraumatic stress disorder, wherein comprising drug solution, drug suspension,
Cubic liquid crystal.
3. the nasal formulations composition for the treatment of posttraumatic stress disorder according to claim 1 or 2, it is characterised in that described
Effective component in drug solution is tetrandrine.
4. temperature-sensitive gel according to claim 1 or 2, it is characterised in that raw material contains 0.1 in parts by weight~
10 parts of PLURONICS F87s, 1~10 part of poloxamer188,15~35 parts of water.
5. it is anti-that raw material contains 0.1~10 part of powder in parts by weight the present invention provides a kind of tetrandrine nasal formulations composition
Own alkali, 30~90 parts of cubic liquid crystals.
6. the preparation side of the nasal formulations composition of the treatment posttraumatic stress disorder described in any one of according to claim 1
Method comprising the step of mixing drug solution, temperature-sensitive gel.
7. treating the preparation side of the nasal formulations composition of posttraumatic stress disorder according to any one of claim 2
Method comprising the step of mixing drug solution, cubic liquid crystal.
8. the preparation method of the nasal formulations composition for the treatment of posttraumatic stress disorder according to claim 4, packet
It includes the following steps: 1) ultrapure water is added at 4 DEG C in the poloxamer188 and PLURONICS F87 that include in nasal formulations composition
Lower sufficiently swelling, continues to add water to full dose, up to temperature-sensitive gel after mixing；2) will include in nasal formulations composition
Drug solution is added in temperature-sensitive gel and mixes to get nasal gel preparation.
9. the preparation method of the nasal formulations composition for the treatment of posttraumatic stress disorder according to claim 5, packet
It includes the following steps: (1) centrifuge tube equipped with glyceryl monooleate being put into 45 DEG C of thermostat water baths and be heated to single oleic
Ester melts completely, ethyl alcohol is added, vortex is uniformly mixed it, as oily phase；(2) drug is added to mix；(3) it draws and is preheated to 45
DEG C deionized water, water droplet is added in oily phase in vortex, after mixing evenly at room temperature place 7 days.
10. the preparation method of the nasal formulations composition for the treatment of posttraumatic stress disorder according to claim 5, packet
It includes the following steps: (1) ethyl alcohol being added in glyceryl monooleate, the container heating water bath of the two will be filled, until glyceryl monooleate
Quan Rongzhi clear transparent solutions；(2) tetrandrine and water are mixed, it is same to put 45 DEG C of heating；(3) centrifuge tube is held to mix in whirlpool
On device, tetrandrine suspension is drawn with 1ml syringe, is added dropwise rapidly, until centrifugation liquid in pipe gradual change stiff, continues to stir
It mixes, continues that surplus solution is added dropwise.
11. nasal formulations composition according to any one of claim 1 to 10 is prepare stress after wound for treating
Purposes in the drug of obstacle.
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