CN108187031A - RhIGF1 is preparing the application in treating fragile X mental retardation drug - Google Patents
RhIGF1 is preparing the application in treating fragile X mental retardation drug Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
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Abstract
The invention discloses rhIGF1 (insulin-like growth factor-1, recombinant human insulin like growth factor 1) to prepare the application in treating fragile X mental retardation drug.The research of the present invention fully demonstrates after the birth of fragile X mental retardation mouse in serum and cerebral hippocampus area type-1 insulin like growth factor (insulin like growth factor 1, IGF1 it is) horizontal to be significantly lower than normal mouse of the same age, the level that can improve its IGF1 is injected intraperitoneally in rhIGF1, reaches the level similar to normal mouse of the same age.RhIGF1 is abnormal to the core symptom such as Development of Hippocampal Neurons of fragile X mental retardation, learning and memory defect, human communication disorders, anxiety and huge testis disease etc., good therapeutic effect is respectively provided with, it is related that these effects with it adjust the excessive activation of IGFR1 AKT S6K signal paths.Clinically be successfully used for the treatment of Rett syndromes and insulin resistance, there is good safety, it is abundant with Research foundation with the medicine that other are being researched and developed compared with, possible research cycle short advantage.
Description
Technical field
The invention belongs to pharmaceutical technology field, specially rhIGF1 is preparing the application in treating fragile X mental retardation drug.
Background technology
Fragile X mental retardation (Fragile X syndrome, FXS) is that a kind of common X-linkage, single-gene are lost
Transmissibility intellectual development disorder disease, male's incidence are about 1/5000, and pole heavy burden is brought to patient home and society.It is main
Clinical manifestation is middle severe intellectual development obstacle, and is often accompanied by autism spectrum disorder (Autism spectrum
Disorders, ASDs), anxiety disorder, epilepsy etc., it is also embodied by huge testis disease in male.Research thinks that its Etiological is due to crisp
Property X intellectual development obstacles gene (Fragile X mental retardation gene, FMR1) 5 ' end three nucleosides of non-translational region
Sour CGG excessive amplifications cause it to methylate, so as to cause fragile X intellectual development obstacle albumen (the Fragile X of its coding
Mental retardation protein, FMRP) missing.FMRP is a polysome correlation rna binding protein, is responsible for
Adjust a variety of expression for playing an important roll albumen to synaptic plasticity, such as PSD-95, Shank1-3 etc..Classical theory thinks,
MGluR5 excessive activations lead to Development of Hippocampal Neurons and plastic sexual abnormality, so as to cause a variety of rows such as FXS intellectual development sluggishnesses
Do not obtain good result to change, but for the treatment of this target spot, it is other it has been found that potential active drugs, such as
Arbaclofen and minocycline etc., few there is also target user in clinical trial, the defects of side effect is apparent.To being at present
Only, the effective means for being clinically directed to FXS still without one, therefore, new medicine and target spot urgently excavate.
Type-1 insulin like growth factor (Insulin-like growth factor 1, IGF1) is one by 70 amino
The growth hormone of acid composition, plays an important roll ontogeny.The mouse projection neuron cell space that studies have shown that IGF1 lacks
Volume, dendron length, Dendritic arborization and synaptic density etc. have apparent exception, so as to lead to the reduction of its cranial capacity.More
Evidence suggests insulin-like growth factor-1 (recombinant human IGF1, rhIGF1) is to Rett syndromes
There is good therapeutic effect and safety with insulin resistance.
Invention content
The object of the present invention is to provide rhIGF1 to prepare the application in treating fragile X mental retardation drug, so as to be fragile X
Syndrome provides a kind of novel drugs.
Including：RhIGF1 is preparing the application in treating fragile X mental retardation drug.
Applications of the rhIGF1 in treatment mammal and/or people's fragile X mental retardation drug is prepared.
Applications of the rhIGF1 in treatment mouse and/or people's fragile X mental retardation drug is prepared.
RhIGF1 treats fragile X mental retardation in preparation by adjusting the excessive activation of IGFR1-AKT-S6K signal paths
Application in drug.
RhIGF1 prepare by adjust the excessive activation of IGFR1-AKT-S6K signal paths treat mammal and/
Or the application in people's fragile X mental retardation drug.
By adjusting the excessive activation of IGFR1-AKT-S6K signal paths to treat mouse and/or people crisp preparing by rhIGF1
Application in property X syndrome drug.
The fragile X mental retardation show as Development of Hippocampal Neurons exception, amentia, human communication disorders, anxiety and/or
Huge testis disease.
Advantages of the present invention includes：
The research of the present invention fully demonstrates horizontal with cerebral hippocampus area IGF1 in serum after fragile X mental retardation mouse is born
Significantly lower than normal mouse of the same age, the level that can improve its IGF1 is injected intraperitoneally in rhIGF1, reaches and normal mouse phase of the same age
As it is horizontal.RhIGF1 is abnormal to the core symptom such as Development of Hippocampal Neurons of fragile X mental retardation, and learning and memory defect is social
Obstacle, anxiety and huge testis disease etc., is respectively provided with good therapeutic effect, these effects adjust IGFR1-AKT-S6K letters with it
The excessive activation of number access is related.Insulin-like growth factor-1 (abbreviation rhIGF1) is clinically successfully used for
The treatment of Rett syndromes and insulin resistance has good safety, has compared with the medicine that other are being researched and developed
Have that Research foundation is abundant, possible research cycle short advantage.
Description of the drawings
Attached drawing is for providing further understanding of the disclosure, and a part for constitution instruction, with following tool
Body embodiment is used to explain the disclosure, but do not form the limitation to the disclosure together.In the accompanying drawings：
Fig. 1 be different disposal group mouse after birth 28 days and 56 days when, noted in rhIGF1 or physiological saline (Vehical)
Penetrate preceding 2h, after injection 2h and for 24 hours serum and hippocampus IGF1 levels detection figure.Wherein A be serum I GF1 testing results, every group 6
A mouse, B be hippocampus IGF1 testing results, every group of 6 mouse.*P<0.05 represents there is statistics compared with WT+Vehicle groups
Learn meaning, #P<0.05 represents that KO+Vehicle groups are compared with statistical significance.
Fig. 2 is after mouse birth 56 days (P56), i.e. after rhIGF1 is injected 6 weeks, detection different disposal is dyed using Golgi
Group hippocampus neuronal development situation map, wherein figure A is separate groups of mice hippocampus neuron dendritic spines aspect graph, B is is recognized
For elongated, the small dendritic spines quantity statistics figure of developmental immaturity, 15 neurons of the every group of data from 5 mouse.*P<
0.05 represents there is statistical significance, #P compared with WT+Vehicle groups<0.05, which represents that KO+Vehicle groups are compared to have, counts
Learn meaning.
Fig. 3 is ability of learning and memory in mice variation testing result figure after evaluation rhIGF1 is handled 6 weeks.A is background fear note
Recall testing result, accounting for total detection time percentage with the stiff time represents；B is condition correlation fear memory testing result, is examined altogether
Surveyed ten times, list here preceding four detections as a result, because of comparison among groups no difference of science of statistics each since the 4th time, every group
There are 8 mouse.*P<0.05 represents there is statistical significance, #P compared with WT+Vehicle groups<0.05 represents KO+Vehicle
Group, which is compared, has statistical significance.
Fig. 4 is mouse social activity preference variation testing result figure after evaluation rhIGF1 is handled 6 weeks.A is test experience mouse flower
Take the exploration time around new mouse；B be test experience mouse within the detection phase to the sniff number of new mouse, every group equal 8
Mouse.*P<0.05 represents there is statistical significance, #P compared with WT+Vehicle groups<0.05 represents KO+Vehicle group phases
Than having statistical significance.
Fig. 5 is mouse anxiety Behavioral change testing result figure after evaluation rhIGF1 is handled 6 weeks.A, B are spacious field experiment knot
Fruit, wherein A have counted different disposal group mouse in the spacious field central area activity time, and B has counted its total activity within the detection phase
Distance.C, D are elevated plus-maze test as a result, wherein C represents residence time of the mouse in open arms, D expression each group mouse
Into open arms and the total degree for closing arm.*P<0.05 represents there is statistical significance, #P compared with WT+Vehicle groups<0.05 table
Show
KO+Vehicle groups, which are compared, has statistical significance.
Fig. 6 is evaluation rhIGF1 before processings (P14) and processing 2 weeks (P28), after 4 weeks (P42) and 6 weeks (P56), mouse
After changes of weight situation map (A) and each group mice behavior detect, the weight statistical chart (B) of testis is isolated.*P<
0.05 represents there is statistical significance, #P compared with WT+Vehicle groups<0.05, which represents that KO+Vehicle groups are compared to have, counts
Learn meaning.
Fig. 7 is the signal of fragile X mental retardation model mice intracerebral hippocampus abnormal activation after evaluation rhIGF1 is handled 6 weeks
The activity change situation map of molecule.A is insulin-like growth factor receptor 1 (IGFR1) in the expression of different disposal group, B
It is AKT phosphorylation level situations of change, C is the downstream molecules S6K phosphorylation level situations of change of mTORC1.*P<0.05 represents
There is statistical significance, #P compared with WT+Vehicle groups<0.05 represents that KO+Vehicle groups are compared with statistical significance.
Specific embodiment
The specific embodiment of the disclosure is described in detail below in conjunction with attached drawing.It should be understood that this place is retouched
The specific embodiment stated is only used for describing and explaining the disclosure, is not limited to the disclosure.
Serum and intracerebral hippocampus IGF1 levels are shown after the studies have shown that fragile X mental retardation model mice birth of inventor
Writing reduces.Accordingly, rhIGF1 is used to prepare the application for the treatment of fragile X mental retardation drug by the present invention for the first time.Through consulting literatures, not
See rhIGF1 to the pharmacological action of fragile X mental retardation patient or animal pattern and the report of therapeutic effect.
The research of the present invention fully demonstrates horizontal with cerebral hippocampus area IGF1 in serum after fragile X mental retardation mouse is born
Significantly lower than normal mouse of the same age, the level that can improve its IGF1 is injected intraperitoneally in rhIGF1, reaches and normal mouse phase of the same age
As it is horizontal.RhIGF1 is abnormal to the core symptom such as Development of Hippocampal Neurons of fragile X mental retardation, amentia (learning and memory
Defect), human communication disorders, anxiety and huge testis disease etc. are respectively provided with good therapeutic effect, these effects adjust IGFR1- with it
The excessive activation of AKT-S6K signal paths is related.
For the level of detection serum and hippocampus IGF1, examined using the mouse IGF1 enzyme-linked immunization (ELISA) of commercialization
Test agent box completes different disposal group mice serum and the horizontal variation of hippocampus IGF1.
To detect influences of the rhIGF1 to fragile X mental retardation model mice abnormal behaviour, using conditioned fear memory experiment
Its learning and memory level is detected, its Social behaviors is detected using three casees experiments, is tested using spacious field experiment and high family's plus maze
Its anxiety behavior is detected, it is found that rhIGF1 is respectively provided with good improvement to these abnormal behaviors of fragile X mental retardation model mice
Effect.
For influences of the detection rhIGF1 to fragile X mental retardation model mice neuronal development exception, commercialization is employed
Golgi staining kits have carried out different disposal group hippocampus of mice area neuron staining analysis, it is found that rhIGF1 can promote
Fragile X mental retardation model mice neuron normal development.
In addition, being had recorded also by the mode weighed, mouse weight changes and rhIGF1 is small to fragile X mental retardation model
The effect of the huge testis disease of mouse finds that rhIGF1 does not influence mouse weight, but significantly reduce fragile X mental retardation model mice
Testicular weight.
The present invention is explained further below by specific embodiment, but the present invention is not limited only in embodiment.
1st, Research idea of the invention and main innovative point are summarized：
The IGF1 levels variation of 28 days and 56 days after present invention confirmation fragile X mental retardation model mice birth first, then
It investigates and gives within continuous 4 weeks rhIGF to model mice behavior and physically different therapeutic effect and pharmacological mechanism.Present invention hair
RhIGF1 treatments in existing 4 weeks significantly improve model mice learning memory disorder, and mood and Social behaviors are abnormal, in addition huge testis disease
Also be improved significantly.Pharmacological mechanism studies have shown that rhIGF1 may be by that IRS1/AKT/mTORC1/S6K is inhibited to lead to
Road is active and plays therapeutic effect.
2nd, specific embodiment：
FMR1KO mouse and WT mouse are taken, is divided into WT+Vehicle, WT+rhIGF1, KO+Vehicle, KO+rhIGF1 tetra-
A group.Intraperitoneal injection rhIGF1 0.25mg/kg or saline is given once daily since 14 days (P14) after birth in mouse
(Vehicle), continuous 6 weeks.Before Behaviors survey, to reduce the tension of animal, every mouse is in daily experimental provision institute
Receive experimental implementation person in laboratory and stroke 3min, continuous three days.After the detection of each mouse, in alcohol washes chest
Wall and ground, in case the remaining smell of animal impacts subsequent experimental.Research contents is as follows：
Embodiment one：Long-term rhIGF1 handles the influence to FMR1KO mice serums and hippocampus IGF1 levels
The present invention has detected 28 days (P28) and 56 days (P56) mice serums and intracerebral learning and memory key after birth first
Core group hippocampus IGF1 is horizontal and whether exogenous intraperitoneal injection rhIGF1 (0.25mg/kg) can adjust mouse blood for a long time
The level of cleer and peaceful hippocampus IGF1.Take different disposal group of the same age be born after 28 and 56 days mouse, on the day of before rhIGF1 injections, note
Rear 2h and for 24 hours is penetrated, heart extracting blood after anaesthetizing respectively stands 30min, and 2000 leave heart 5min, take serum, and -80 DEG C of freezings are to be checked.
Mouse breaks end rapidly after putting to death takes brain, is rapidly separated hippocampus, weighs respectively, by tissue weight (mg)：Lysate volume=1:30
Lysate is added in, prepares brain tissue sample, -80 DEG C of freezings are to be checked.Using ELISA kit, blood is detected according to its operating procedure
IGF1 is horizontal in clear and hippocampal tissue sample.
The result is shown in Figure 1, by the result in Fig. 1 it is found that in P28 and P56 mouse, WT+Vehicle mice serums and hippocampus
IGF1 levels are all remarkably higher than KO+Vehicle mouse, and with advancing age, IGF1 levels are in WT and KO mouse in decline
Trend.2h after intraperitoneal injection rhIGF1, two mouse genotypes serum dramatically increase before being compared with hippocampus IGF1 levels,
But the IGF1 levels of WT mouse are higher than KO mouse.After for 24 hours, the horizontal phases of IGF1 of KO+rhIGF1 mouse and WT+Vehicle mouse
Than the trend being reduced, but no difference of science of statistics.After these are the result shows that exogenous abdominal cavity gives rhIGF1, it can be mended in 2h
It fills KO mice serums and hippocampus IGF1 is insufficient, the IGF1 with matching needed for WT mouse can be provided for KO mouse.
Embodiment two：Long-term rhIGF1 handles the influence to FMR1KO mouse hippocampal neuron forms
A large amount of cards are it was demonstrated that the immature dendritic spines density showed increased of FMR1KO hippocampus of mice area cone neurone.This
Mouse 5 of the same age is taken in invention for every group, is washed after directly putting to death, and requires to operate according to Golgi staining kits specification, at random
3 neurons are selected, totally 15 neurons, select the dendrons at different levels in 100 μm of cell space, Image is used at interval of 10 μm
J softwares record total dendritic spines and slender type (thin) the dendritic spines quantity for being considered immature long-neck, microcephaly on 50 μm of dendrons altogether.
As a result Fig. 2 is seen, by the result in Fig. 2 it is found that birth 56 days (P56), KO+Vehicle group hippocampus of mice dentate fascias
Neuron elongated dendritic spines density in area's is noticeably greater than WT+Vehicle group mouse.It is long-term to give intraperitoneal injection rhIGF1 group KO mouse
Neuron elongated dendritic spines density in hippocampal dentate area is significantly less, hence it is evident that less than KO+Vehicle group mouse, with WT+rhIGF1
The density of group mouse is suitable.In addition, long-term rhIGF1 processing is not also to the elongated dendritic spines density of WT adult mouse dentate gyrus area neuron
It makes a significant impact.These the result shows that rhIGF1 long-term disposals can to substantially reduce the elongated dendritic spines of FMR1KO hippocampus of mice close
Degree is prompted it is possible that improving FMR1KO learning and memory of little mouse level.
Embodiment three：Long-term rhIGF1 handles the influence to FMR1KO mouse anxiety sample behaviors
A large amount of evidences show that FMR1KO mouse show apparent anxiety-like behavior, while have the performance of hyperactivity shape.This
Invention is using internationally recognized classical spacious field experiment (Open field test, OFT) Elevated plus-maze (Elevated
Plus-maze test, EPM) detect mouse anxiety sample behavior, mainly using mouse to spacious field central area and elevated plus fan
Conflict nervous psychology between the fear in palace open arms area and novel exploration desire to judge the anxiety degree of mouse, mouse is more burnt
Consider, the residence time in spacious field central area and high family's plus maze open arms area is shorter.Mouse movement activity is stronger simultaneously, difference
Move distance in two experimental provisions is longer, more into arm number.
Spacious field experimental provision is made of spacious field reaction chamber and data acquisition and analysis system, and spacious field reaction chamber is one equipped with light
According to the sound proof box with air interchanger, size is 30cm long, 30cm wide, 30cm high.Surrounding and bottom are transparent plexiglas
Square box, has a visual field that can cover the digital camera head inside entire spacious field above cabinet interior by totally four, before experiment,
First set relevant parameter, during experiment, mouse is put in reaction chamber central area, while opens the activity feelings of camera record mouse
Condition, each chest put a mouse every time.Mouse is allowed freely to live 15min, identifies track, record mouse is on reaction chamber total activity road
Journey and central area activity time.We define 15 × 15cm of experimental box bottom middle2Region is central area, accounts for entire area
A quarter.
Elevated plus-maze device by two open arms (25cm × 8cm × 0.5cm) and two close arm (25cm × 8cm ×
12cm) the cross-shaped composition of journey, intersection region form halfpace, and length and width are 8cm, whole device 50cm from the ground, and
It is surrounded to reduce external interference with the door curtain made of cloth.Video-frequency monitor be placed in device center top about 1.5m, and with mouse ten on computer
Word Y-maze test software is connected.During experiment, open arms is defined on software first, closes arm and middle section, the ginseng of each arm is set
Number adjusts camera position.During detection, mouse head towards open arms is put in device intermediate region, and (hereafter every animal is put into same
Position), experimenter tenses the door curtain made of cloth immediately, leaves test section, starts to detect, timing 5min, and software records open and close arm enters
It number, residence time and into arm total degree, makes an appraisal after statistical analysis to the anxiety level of mouse.WT+Vehicle, WT+
RhIGF1, KO+Vehicle, KO+rhIGF1
As a result Fig. 5 is seen, from the result in Fig. 5：
Figure A and figure B spacious field experimental results show that in P56 mouse, KO+Vehicle mouse are small compared to WT+Vehicle
Mouse significantly reduces in the activity time of spacious field central area, for being found in the detection of its mobility, KO+Vehicle mouse fortune
It moves apart from showed increased.After chronic rhIGF1 processing, KO mouse rise appreciably in the activity time of central area, and move distance
But it significantly reduces.The chronic rhIGF1 processing of these result preliminary proofs can extend the FMR1KO mouse central movable times, reduce
Its move distance.Prompting rhIGF1 may have the function of KO mouse angst resistance effect and adjust hyperactivity.
Scheme C and the plus maze detection display of figure D high families, in P56 mouse, KO+Vehicle mouse are compared to WT+
Vehicle mouse significantly reduce in the open arms residence time of Elevated plus-maze, for being found in the detection of its mobility, KO
+ Vehicle mouse are in open arms and close arm and enter on total degree and are significantly higher than WT+Vehicle mouse.In chronic rhIGF1 processing
Afterwards, KO mouse significantly extend its residence time in open arms, while reduce the total degree into opening and closing arm.These results are demonstrate,proved
Bright chronic rhIGF1 processing can extend the FMR1KO mouse open arms residence times, reduce it into arm total degree.Further prompting
RhIGF1 has the function of angst resistance effect in FMR1KO mouse and adjusts hyperactivity.
Example IV：Long-term rhIGF1 handles the influence to FMR1KO learning and memory of little mouse level
For fragile X mental retardation patient, most crucial symptom is feeblemindedness, shows that ability of learning and memory is serious
In terms of shortage.The present invention carries out rhIGF1 using conditioned fear memory experiment in terms of KO ability of learning and memory in mice improvement
Further investigation.Experiment carries out in the case that shocks by electricity, and conditioning stimulus (Conditioned stimulus, CS) is to continue 15s, 80dB
White noise, unconditioned stimulus (Unconditioned stimulus, US) be continue 0.5s, the foot shock of 0.7mA.Experiment
1st day, animal is exposed in experimental box after adapting to 120s and is given respectively persistently at the 120th, 180,240,300 and 360s
The white noise Sound stimulat of 30s gives the foot shock of lasting 2s simultaneously in each last 2s of white noise Sound stimulat.Entire Therapy lasted
400s observes the stiff time (freezing time) of animal by camera.It tests the 2nd day, animal is placed in experimental box
120s is not to any sound or electro photoluminescence, and the deadlock that animal is observed by camera lives the time, to evaluate background fear memory
(contextual fear memory).It tests the 3rd day, after experimental box background is changed completely, mouse is put into experimental box,
0-30s is adaptation time, and 30s gives the white noise stimulation 10s identical with the training stage, gives white noise every 60s later
10s is stimulated, totally 10 times, records stiff time of each mouse, with related (sound) fear memory (the cued fear of evaluation condition
memory).The stiff time refers to animal other than respiratory movement, the wholly off state of other somatic movements of whole body, in the detection phase
Interior accounting is bigger, illustrates that its ability of learning and memory is stronger.
As a result see Fig. 3, by the result in Fig. 3 it is found that in the detection of background fear, KO+Vehicle mouse are within the detection phase
Dead time percentage be substantially less than WT+Vehicle mouse, showing the learning and memory abilities of KO mouse, to be substantially less than WT small
Mouse.And the dead time percentage of the KO mouse of rhIGF1 processing significantly increases, without statistics compared with WT+Vehicle mouse
Difference dramatically increases compared with KO+Vehicle mouse.The result shows that the background that rhIGF1 processing can improve KO mouse is related
Fear memory.In related (i.e. sound) fear memory detection of condition, KO+Vehicle mouse are motionless within 10 seconds detection phases
Time is held essentially constant, substantially less than WT+Vehicle mouse.And the dead time of rhIGF1 processing mouse dramatically increases,
It is significantly higher than KO+Vehicle mouse, slightly below WT+Vehicle mouse, but no difference of science of statistics.In 10 seconds detection phases of the 4th
No difference of science of statistics between the dead time of interior each group shows that mouse has adapted to new stimulus modality again.The above result shows that
RhIGF1 processing can significantly improve the fear memory ability of KO mouse, prompt core diseases of the rhIGF1 to fragile X mental retardation
Shape-feeblemindedness may have improvement result.
Embodiment five：Long-term rhIGF1 handles the influence to FMR1KO mouse Social behaviors
A large amount of cards are it was demonstrated that FMR1KO mouse and fragile X mental retardation patient have autism sample symptom.The present invention uses three
Case experimental evaluation mouse social interaction behavior.Experimental provision is made of three organic glass casees and video track monitoring system, in
Between case it is smaller (8 × 22.5 × 30.5cm), both sides box sizes are equal (24.5 × 22.5 × 30.5cm), and it is small to set one between each case
Door (5 × 8cm).Experiment is divided into two stages, and different disposal mouse is individually tested successively.First stage is the laundering period,
Two empty wire mesh cages (diameter 8cm) are then put into experimental box, stop 10min by mouse free shuttling 5min in experimental box,
Close wicket.Second stage is social preference detection-phase, and wooden mouse and one are placed respectively at random in two empty wire mesh cages
Experiment mice opens wicket, video track monitoring systematic perspective from not in contact with the male mice of the same age but different genotype crossed
It examines and records experiment mice in 5min and time and sniff number are explored in new mouse case, to investigate the positive of mouse social activity
Property.
As a result Fig. 4 is seen, by the result in Fig. 4 it is found that exploration time of the KO+Vehicle mouse around new mouse is notable
It reduces, while its sniff number is also significantly lower than WT mouse.Compared with KO+Vehicle mouse, the KO mouse pair of rhIGF1 processing
The sniff number showed increased of new mouse, exploring the time around new mouse also significantly extends, these indexs and WT+Vehicle
Mouse is compared to no statistics difference.These statistics indicate that, rhIGF1 significantly extends exploration time of the FMR1KO mouse to new mouse
With sniff number, prompt rhIGF1 processing that can improve the sociability of FMR1KO mouse, improve its autism sample behavior.
Embodiment six：The influence of long-term rhIGF1 processing huge testis disease to FMR1KO mouse
All experiment mices after experiment carefully detach testis and weigh immediately.
FMR1KO mouse are exactly huge testis disease there are one apparent sign.As a result Fig. 6 is seen, by the result in Fig. 6 it is found that WT
+ Vehicle mouse and KO+Vehicle mouse testis weight are there are difference, and the FMR1KO mouse testis of rhIGF1 processing
Ball weight is compared with WT mouse, no statistics difference, and substantially less than it compares KO mouse.In addition, rhIGF1 processing is small to each group
The weight of mouse does not influence.These evidences show that rhIGF1 can correct the huge testis symptom of FMR1KO mouse, in FMR1KO
Better effects are obtained in the huge testis disease treatment of mouse.
Embodiment seven：Long-term rhIGF1 processing is to the molecular mechanism of the therapeutic effect of FMR1KO mouse
Evidence suggests FMR1KO mouse ImTORC1 signal path excessive activations.The present invention uses westernblot methods,
MTORC1 signal paths are detected, to investigate molecular mechanism of the long-term rhIGF1 processing to the therapeutic effect of FMR1KO mouse
As a result see Fig. 7, by the result in Fig. 7 it is found that in P56 mouse, KO+Vehicle hippocampus of mice area IGFR1 levels with
And AKT, S6K phosphorylation level significantly increase, the IGFR1 that rhIGF1 processing significantly reduces KO mouse is horizontal, reduces simultaneously
AKT, S6K phosphorylation level, the no difference of science of statistics compared with WT+Vehicle mouse.These results prove that rhIGF1 can inhibit
The excessive activation of FMR1KO mouse IGFR1-AKT-S6K signal paths prompts rhIGF1 in terms of the symptom improvement of KO mouse
Effect may be by adjusting the excessive activation of IGFR1-AKT-S6K signal paths and generating.
To sum up, from after birth 14 days, continue 6 weeks, daily 0.25mg/kg rhIGF1 processing is to the crisp of FMR1KO mouse
Property X syndrome core symptom such as Development of Hippocampal Neurons it is abnormal, learning and memory defect, human communication disorders, anxiety and huge testis
Disease etc. is respectively provided with good improvement result, this effect may be by adjusting the excessive of IGFR1-AKT-mTORC1 signal paths
It activates and generates, it was demonstrated that rhIGF1 can be prepared into the application for the treatment of fragile X mental retardation drug.
Claims (7)
1.rhIGF1 is preparing the application in treating fragile X mental retardation drug.
Applications of the 2.rhIGF1 in treatment mammal and/or people's fragile X mental retardation drug is prepared.
Applications of the 3.rhIGF1 in treatment mouse and/or people's fragile X mental retardation drug is prepared.
4.rhIGF1 treats fragile X mental retardation medicine in preparation by adjusting the excessive activation of IGFR1-AKT-S6K signal paths
Application in object.
5.rhIGF1 prepare by adjust the excessive activation of IGFR1-AKT-S6K signal paths treat mammal and/or
Application in people's fragile X mental retardation drug.
6.rhIGF1 treats mouse and/or people's fragile X in preparation by adjusting the excessive activation of IGFR1-AKT-S6K signal paths
Application in syndrome drug.
7. the application described in claim 1-6 any claims, which is characterized in that the fragile X mental retardation shows as sea
Horse neuronal development exception, amentia, human communication disorders, anxiety and/or huge testis disease.
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| CN114042149B (en) * | 2021-12-03 | 2023-06-16 | 中国人民解放军空军军医大学 | D-Ala 2 Use of GIP for the preparation of a medicament for the treatment of anxiety disorders |
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| CN101820896A (en) * | 2007-06-08 | 2010-09-01 | 麻省理工学院 | The IGF that is used for the treatment of Rett syndrome and synaptic disorders |
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| CN114042149B (en) * | 2021-12-03 | 2023-06-16 | 中国人民解放军空军军医大学 | D-Ala 2 Use of GIP for the preparation of a medicament for the treatment of anxiety disorders |
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