CN106854233A - One species peptide and its preparation method and application - Google Patents
One species peptide and its preparation method and application Download PDFInfo
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- CN106854233A CN106854233A CN201710124106.6A CN201710124106A CN106854233A CN 106854233 A CN106854233 A CN 106854233A CN 201710124106 A CN201710124106 A CN 201710124106A CN 106854233 A CN106854233 A CN 106854233A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
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- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
- G01N33/6896—Neurological disorders, e.g. Alzheimer's disease
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Abstract
The present invention provides species peptide and its preparation method and application, and the class peptide includes following subunit：Ethylenediamine (I), piperonyl amine (II), beta Alanine (III), 1 naphthylamines (IV) and cysteine (V), class peptide symthesis are simple and stronger with α synapse nucleoprotein binding abilities, examination effectively can be carried out to patient PD and normal human serum by the α synapse nucleoproteins in serum, new liquid biopsy method and thinking is provided to diagnose and monitoring Parkinson's.
Description
Technical field
The invention belongs to medicine and pharmacology technical field, it is related to species peptide and its preparation method and application.
Background technology
Parkinson's (Parkinson ' s Disease, PD) are a kind of very common nervous system degeneration diseases, occurred frequently
With Elderly patients, its average age of onset is or so 60 one full year of life.Up to now, the PD illness rates of China's over-65s crowd
1.7% (but this numeral may be artificial relatively low, because many patients in rural area are never diagnosed), and big portion are reached
The Parkinsonian for dividing is Sporadic cases, and the only patient less than 10% has the household heredity factors that can be examined.Parkinson's
Main pathology lesion be that the denaturation of substantia nigra of midbrain dopamine (dopamine, DA) serotonergic neuron is dead, line shape is caused therefrom
Body DA contents are substantially reduced and caused a disease.Although but have numerous researchs on Parkinson's at present, its definite cause of disease mesh
Preceding still unclear, inherent cause, environmental factor, age ageing and oxidative stress etc. may participate in PD dopaminergic neurons
Denaturation death process.British physician James Parkinson are described in detail to this disease first within 1817, its clinical table
Now mainly include static tremor, bradykinesia, myotonia and posture gait disorder, at the same patient can with depressed, constipation and
The non-motor symptoms such as sleep-disorder.The diagnosis of Parkinson's relies primarily on medical history, clinical symptoms and sign.General auxiliary examination
Change more without exception.Drug therapy is the topmost treatment means of Parkinson's.Dopar is still maximally effective medicine.
Operative treatment is one kind effectively supplement of drug therapy.Rehabilitation, psychotherapy and good nursing also can be to a certain degree
Upper improvement symptom.Although the treatment means of application can only improve symptom at present, it is impossible to prevent the progress of the state of an illness, cannot also cure disease
Disease, but effective treatment can significantly improve the quality of life of patient.The life expectancy of PD patient is with general population without significant difference.
But China, used as the big country of population in the world first, the social phenomenon of current aging is increasingly severe, therefore Parkinson's turn into
China Today society and medical treatment have to the significant problem for facing.
The pathological change that Parkinson's are protruded is that the denaturation of substantia nigra of midbrain dopamine (dopamine, DA) serotonergic neuron is dead
Die, striatum DA level conspicuousness reduce and black substance remaining neuron kytoplasm in there is eosinophilic inclusion, i.e. Lewy body
(Lewy body).At least more than 50%, striatum DA level subtracts substantia nigra dopaminergic neuron death during clinical symptoms occurs
Less more than 80%.In addition to dopaminergic system, the non-dopaminergic system of Parkinsonian also has obvious impaired.Such as
The cholinergic neuron of Meynert basal nucleis, the noradrenergic neuron of locus coeruleus, the serotonin energy of brain stem nuclei of median raphe
Neuron, and cerebral cortex, brain stem, spinal cord and periphery autonomic nerves system neuron.Striatal Dopamine Content shows
Write decline closely related with the appearance of Parkinson's motor symptoms.Midbrain-limbic system and midbrain-Cortical systems dopamine concentration
Significantly reduce with Parkinsonian that hypophrenia, the disturbance of emotion etc. occur closely related.Alpha-synapse nucleoprotein is that one kind exists
Central nervous system presynaptic and the soluble protein of core week expression, its pathogenesis and correlation function barrier with Parkinson's
Hinder closely related, be the main component of Lewy body.
Research finds that alpha-synapse nucleoprotein is normal, there is dynamic equilibrium between false folding and its oligomerization, when this flat
Weighing apparatus is broken rear fibrillation and is gathered into macromolecular, insoluble fine fibre rapidly；Alpha-synapse nucleoprotein is in different influence factors
Under can show variform perhaps, including unfold state, dissolving forecourt kenel, alpha-helix state (film combination), β-piece attitude, dimer
State, oligomerization figure and insoluble unformed shape and fiber state；Structural change caused by the point mutation of alpha-synapse nucleoprotein,
Concentration (the pH value of the increase of intracellular content, the accumulation of a large amount of protein moleculars, the truncation of structure sequence, intracellular anion and salt
Change), neurotoxic molecule (heavy metal, organic solvent, carbon monoxide, MPTP, insecticide and herbicide), translation after repair
Decorations (oxidation, phosphorylation and nitration) etc. can promote alpha-synuclein aggregation to form indissoluble fiber；It is presently believed to widow
The alpha-synapse nucleoprotein of poly- state most cytotoxicity.
The diagnosis of Parkinson's relies primarily on medical history, clinical symptoms and sign.According to the hidden spy for attacking onset, being gradually in progress
Point, one side is involved and then is developed to offside, is shown as static tremor and is slow in action, and excludes atypia Parkinson's sample symptom
Clinical diagnosis can be made.Diagnosis is effectively then more supported to Dopar treatment.Being as good as conventional blood, examination of cerebrospinal fluid more
Often.Also atypism changes for head CT, MRI.Can find that PD patient has hyposphresia more than olfactometry.Using 18F- DOPA as showing
Track agent row DOPA capture functions PET imagings can show that the synthesis of dopamine mediator is reduced.With 125I- β-CIT, 99mTc-TRODAT-1
Can show that DAT quantity is reduced as tracer row DAT (DAT) functional image, in disease early stage even Patients with Subclinical
Can show reduces, and can support diagnosis.It is not yet conventional to carry out but this inspection fee is more expensive.
Since 1997 it is found that alpha-synapse nucleoprotein is mutated with familial Parkinson's disease gene (SCNA) and Louis is small
Since the main component of body is relevant, focus is just become in its Molecular pathogenesis in Parkinson's, we have invented one
Species peptide with alpha-synapse nucleoprotein content in easier, sensitiveer, more inexpensive, hurtless measure method detection blood diagnosing and
Monitoring PD.Class peptide (peptoid) is the foldamers similar to polypeptide structure with N-substituted glycinic acid as unit.It can
To be folded into the functional unit of high bioactivity and high specific, and its component units is more more rich than polypeptide, and is resistant to egg
White enzyme；Therefore, class peptide compounds have good bioactivity and chemical property.
Therefore, it is this area in a kind of reagent that can be used in Parkinson's detection, diagnosis and/or monitoring of this area exploitation
Problem demanding prompt solution.
The content of the invention
In view of the shortcomings of the prior art, it is an object of the invention to provide species peptide and its preparation method and application.This
The described class peptide symthesis of invention are simple and stronger with alpha-synapse nucleoprotein binding ability, can effectively by the α in serum-
Synapse nucleoprotein to patient PD and normal human serum carries out examination.
To reach this goal of the invention, the present invention uses following technical scheme：
On the one hand, the present invention provides a species peptide, and the class peptide includes following subunit：Ethylenediamine (I), piperonyl amine
(II), Beta-alanine (III), naphthalidine (IV) and cysteine (V) subunit.
In the present invention, the structure of the donor of each subunit of the class peptide is as follows：
Preferably, the sequence of the subunit of the class peptide is I-IV-III-II-I-I-V.
Preferably, in the present invention, the class peptide has following structure：
In the present invention, the class peptide with structure as described above is stronger with alpha-synapse nucleoprotein binding ability, can be effective
Ground carries out examination by the alpha-synapse nucleoprotein in serum to Parkinsonian and normal human serum.
Second aspect, the present invention provides a kind of preparation method of class peptide as described above, and the preparation method passes through solid phase
Synthetic method synthesizes.
Preferably, the preparation method is comprised the following steps：
(1) according to the subunit order of connection of class peptide, first subunit of the class peptide is connected on solid phase carrier；
(2) bromoacetic acid is reacted under the activation of activator with the amino of first subunit being connected on solid phase carrier
Form amido link；
(3) product that second donor of subunit of class peptide is obtained with step (2) is reacted, is substituted off bromine former
Son, completes second connection of subunit；
(4) connection of bromoacetic acid and follow-up subunit is then repeated, until completing the connection of all subunits；
(5) the class peptide that obtains will be synthesized from solid phase carrier it will be cleaved and will obtain the class peptide.
Preferably, step (1) described solid phase carrier is Rink amide AM resins.
Preferably, step (1) is described is connected to solid phase on solid phase carrier before first subunit of the class peptide
Carrier carries out swelling.
It is when solid phase carrier is Rink amide AM resins, its is swelling, and cause Rink using hexahydropyridine deprotection
Amide AM resins expose amino.
Preferably, during first subunit of the class peptide is connected on solid phase carrier, in condensing agent and work
Carried out in the presence of agent.
Preferably, the condensing agent is the tetramethylurea hexafluorophosphoric acids of 2- (3'-N- oxos-BTA) -1,1', 3,3'-
In salt, O- BTAs-N, N, N', N'- tetramethylurea tetrafluoro boric acid or I-hydroxybenzotriazole any one or at least
Two kinds of combination.
Preferably, the activator is N- methylmorpholines.
Preferably, step (2) described activator is N, N'- DICs or dicyclohexylcarbodiimide.
Preferably, the temperature of step (2) described reaction be 20-40 DEG C, such as 20 DEG C, 22 DEG C, 25 DEG C, 28 DEG C, 30 DEG C, 32
DEG C, 35 DEG C, 37 DEG C, 39 DEG C or 40 DEG C.
Preferably, the time of step (2) described reaction be 20-100min, such as 20min, 25min, 30min, 35min,
40min, 50min, 60min, 70min, 80min, 90min or 100min.
The donor described in step (3) is the compound for referring to provide the class peptide subunit, and for example cysteine is sub-
The donor of unit is cysteine, and the donor of Beta-alanine subunit is Beta-alanine, the like.
Preferably, the temperature of step (3) described reaction be 20-40 DEG C, such as 20 DEG C, 22 DEG C, 25 DEG C, 28 DEG C, 30 DEG C, 32
DEG C, 35 DEG C, 37 DEG C, 39 DEG C or 40 DEG C.
Preferably, the time of step (3) described reaction be 50-150min, such as 50min, 55min, 60min, 65min,
70min, 80min, 90min, 100min, 120min, 140min or 150min.
Repeat the connection i.e. repeat step (2) of bromoacetic acid and follow-up subunit described in step (4) in the present invention
With step (3), the subunit for only connecting is follow-up subunit.
Preferably, the decomposition agent for being used during step (5) described cracking includes the following composition of mass percent：95% trifluoro
Acetic acid, 2.5% ultra-pure water and 2.5% tri isopropyl silane.
Preferably, the group that the coupled reaction can be will not participate in the preparation process of the polypeptide sub-units is carried out
Radical protection, to ensure the accuracy of connection site, makes reaction more accurately and successfully carry out, and then completes all sub- single
After the connection of unit, then carry out deprotection deprotection base.
On the other hand, the present invention provides a kind of amyloid detection agent, and the amyloid detection agent is included as above
Described class peptide.
Class peptide of the present invention can be as the detection agent of detection amyloid, or conduct detection amyloid
Detection agent a kind of composition.
Preferably, the amyloid is alpha-synapse nucleoprotein.
On the other hand, the present invention provides a kind of pharmaceutical composition, and described pharmaceutical composition includes class peptide as described above.
Preferably, described pharmaceutical composition also includes pharmaceutically acceptable auxiliary material.
Preferably, the pharmaceutically acceptable auxiliary material is excipient, diluent, carrier, flavor enhancement, adhesive or filling
In agent any one or at least two combination.
On the other hand, detection, diagnosis are being prepared and/or is being supervised the invention provides class peptide as described above or pharmaceutical composition
Application in the medicine of the control disease related to alpha-synapse nucleoprotein.
Preferably, the disease is Parkinson's.
Compared with prior art, the invention has the advantages that：
(1) class peptide of the invention is stronger with alpha-synapse nucleoprotein binding ability, is obtained from primitive resonance technique by surface etc.
To class peptide of the invention and alpha-synapse nucleoprotein binding kineticses constant in equilibrium dissociation constant KD be 10-8Mol/L
The order of magnitude；
(2) detect that class peptide is contrasted to patient PD and normal human blood signal intensity from primitive resonance technique using surface etc.,
Can find substantially differentiate patient and normal person using such peptide；
(3) the PD diagnostic techniques based on such peptide can realize noninvasive unmarked quick diagnosis；
(4) class peptide symthesis of the invention are simple, efficiency high, low cost.
Brief description of the drawings
Fig. 1 is class peptide molecule of the invention and concentration is 2.632 μM, 1.316 μM, 0.658 μM, 0.329 μM of alpha-synapse
The result figure from primitive resonance detection such as surface that nucleoprotein is combined；
Fig. 2 is the test result figure of class peptide of the invention and alpha-synapse nucleoprotein specific binding；
Fig. 3 is class peptide of the invention to patient PD and the contrast effect figure of normal human serum signal；
Fig. 4 is the sensitivity test result figure that class peptide of the invention detects serum in PD diagnostic systems.
Specific embodiment
Technical scheme is further illustrated below by specific embodiment.Those skilled in the art should be bright
, the embodiment be only to aid in understand the present invention, be not construed as to concrete restriction of the invention.
Embodiment 1
In the present embodiment, class peptide is synthesized by solid phase subunit synthetic method, specifically includes following steps：
(1) by Rink amide AM resins (substitution level 0.3mmol/g) it is swelling after use hexahydropyridine deprotection, will be partly
Cystine mixes with the tetramethylurea hexafluorophosphate equimolars of 2- (3'-N- oxos-BTA) -1,1', 3,3'-, in N- first
It is coupled under the activation of base morpholine.
(2) in 2M bromoacetic acids and 3.2M N, N '-DIC (DIC) being added into Rink amideAM resins,
30min is reacted at 37 DEG C, by the aminoacylates of resinous terminal；
(3) add 2M primary amine and react 90min at 37 DEG C, bromine atoms are replaced by nucleophilic substitution, complete one
The synthesis of individual subunit；
(4) repeat step (2) and (3) are until complete the synthesis of its counit；
(5) it is to be synthesized finish after, side chain protecting group is removed, and uses 95% trifluoroacetic acid, 2.5% ultra-pure water, 2.5%
From resin be cleaved class peptide standby by tri isopropyl silane.
The molecular formula of the class peptide that the present embodiment is prepared is as follows：
Embodiment 2
In the present embodiment, class peptide is synthesized by solid phase subunit synthetic method, specifically includes following steps：
(1) by Rink amide AM resins (substitution level 0.3mmol/g) it is swelling after use hexahydropyridine deprotection, will be partly
Cystine mixes with O- BTAs-N, N, N', N'- tetramethylurea tetrafluoro boric acid equimolar, in the work of N- methylmorpholines
It is coupled under change.
(2) in 2M bromoacetic acids and 3.2M N, N '-DIC (DIC) being added into Rink amideAM resins,
60min is reacted at 25 DEG C, by the aminoacylates of resinous terminal；
(3) add 2M primary amine and react 50min at 25 DEG C, bromine atoms are replaced by nucleophilic substitution, complete one
The synthesis of individual subunit；
(4) repeat step (2) and (3) are until complete the synthesis of its counit；
(5) it is to be synthesized finish after, side chain protecting group is removed, and uses 95% trifluoroacetic acid, 2.5% ultra-pure water, 2.5%
From resin be cleaved class peptide standby by tri isopropyl silane.The class peptide of synthesis has molecular structure as described in Example 1.
Embodiment 3
In the present embodiment, class peptide is synthesized by solid phase subunit synthetic method, specifically includes following steps：
(1) by Rink amide AM resins (substitution level 0.3mmol/g) it is swelling after use hexahydropyridine deprotection, will be partly
Cystine mixes with I-hydroxybenzotriazole equimolar, is coupled under the activation of N- methylmorpholines.
(2) it is anti-at 40 DEG C in 2M bromoacetic acids and 3.2M dicyclohexylcarbodiimides being added into Rink amide AM resins
20min is answered, by the aminoacylates of resinous terminal；
(3) add 2M primary amine and react 150min at 20 DEG C, bromine atoms are replaced by nucleophilic substitution, complete one
The synthesis of individual subunit；
(4) repeat step (2) and (3) are until complete the synthesis of its counit；
(5) it is to be synthesized finish after, side chain protecting group is removed, and uses 95% trifluoroacetic acid, 2.5% ultra-pure water, 2.5%
From resin be cleaved class peptide standby by tri isopropyl silane.The class peptide of synthesis has molecular structure as described in Example 1.
Embodiment 4
In the present embodiment, the binding ability between class peptide and alpha-synapse nucleoprotein is investigated, specific method is as follows：
Using the specific step of binding ability between surface plasma resonance image-forming technical testing class peptide and alpha-synapse nucleoprotein
It is rapid as follows：
(1) class peptide is dissolved into ddH2In O, concentration is 1-1000 μM；
(2) by sample spot on a SPRi chip surface, every kind of sample repeats 3 points, after 4 DEG C are incubated 12 hours, uses
10X PBS, 1X PBS, ultra-pure water cleaning is dry.Then chip is closed 30 minutes with 1M Monoethanolaminium chlorides, then uses ultra-pure water
Cleaning 5 times, is finally dried up with clean nitrogen；
(3) chip is arranged on SPRi instruments, determines SPRi angles and adjust to optimal optical position, in detection zone choosing
Coherent detection point, including sample spot and blank spot are taken, setting experimental flow rate is 2 μ L/s；
(4) selection PBS passes sequentially through concentration for buffer solution is passed through after flow cell to baseline stability for 2.632 μM, 1.316 μ
M, 0.658 μM, 0.329 μM detected that binding time is 300 seconds, and Dissociation time is 300 seconds, is passed through phosphoric acid between each concentration
Lived again.
(wherein △ AU are for reflecting binding signal intensity in surface plasma resonance image-forming to testing result such as Fig. 1
Unit, is a kind of dimensionless unit) shown in, through fitting, equilibrium dissociation constant KDIt is 6.18 × 10-8Mol/L, this shows class peptide
There is at a relatively high affinity level with alpha-synapse nucleoprotein.
Embodiment 5
In the present embodiment, class peptide is tested the specificity that alpha-synapse nucleoprotein is combined, method is as follows：
Using the specificity of surface plasma resonance image-forming technical testing class peptide combination alpha-synapse nucleoprotein, specific steps
It is as follows：
(1) with embodiment 4, same SPRi chips are made, installed on SPRi instruments, determine SPRi angles and regulation to
Optimal optical position, coherent detection point, including sample spot and blank spot are chosen in detection zone, and setting experimental flow rate is 2 μ L/s；
(2) PBS is selected for buffer solution is passed through after flow cell to baseline stability the alpha-synapse for passing sequentially through that concentration is 2.632 μM
Nucleoprotein (a-syn), human serum albumins (HSA), IgG, transferrins (TRF), IgM are combined test, and binding time is
300 seconds, Dissociation time was to be passed through phosphoric acid between 300 seconds, each concentration to be lived again.
Testing result is as shown in Fig. 2 the combination of class peptide of the invention and alpha-synapse nucleoprotein has specificity higher.
Embodiment 6
In the present embodiment, method is as follows to be detected to serum signal using class peptide：
Using surface plasma resonance image-forming technical testing class peptide, detection PD patients serums are specific with normal human serum
Step is as follows
(1) with embodiment 4, same SPRi chips are made, installed on SPRi instruments, determine SPRi angles and regulation to
Optimal optical position, coherent detection point, including sample spot and blank spot are chosen in detection zone, and setting experimental flow rate is 2 μ L/s；
(2) after selection PBS is passed through flow cell to baseline stability for buffer solution, the blood of different patients and normal person is each led into
Clear dilution (1:5000), Dissociation time is 300 seconds, and each sample room is passed through phosphoric acid and Proteinase K is lived again.
Testing result is as shown in figure 3, be in inhomogeneity peptide concentration lg values (i.e. lgCPDI2) following table surface plasma resonance into
The signal bond strength of picture, from the figure 3, it may be seen that when class peptide concentration is higher, can substantially distinguish patient PD and normal person (normal).
Embodiment 7
In the present embodiment, the test of PD diagnostic system sensitivity is carried out using class peptide, method is as follows：
Using the sensitivity of surface plasma resonance image-forming technical testing PD diagnostic systems detection serum, specific steps
It is as follows：
(1) with embodiment 4, same SPRi chips are made, installed on SPRi instruments, determine SPRi angles and regulation to
Optimal optical position, coherent detection point, including sample spot and blank spot are chosen in detection zone, and setting experimental flow rate is 2 μ L/s；
(2) after selection PBS is passed through flow cell to baseline stability for buffer solution, the blood of different patients and normal person is each led into
Clear dilution, diluted concentration is respectively 1:2000、1:4000、1:8000、1:16000、1:32000, binding time is 300 seconds,
Dissociation time is 300 seconds, and each sample room is passed through phosphoric acid and Proteinase K is lived again.
Testing result is as shown in figure 4, serum-dilution ratio is less than or equal to 1:When 8000, can substantially distinguish patient PD with
Normal person, illustrating it has high sensitivity.
In sum, class peptide of the invention is a kind of alpha-synapse nucleoprotein high sensitivity high specific in serum to be combined
Class peptide, new liquid biopsy method and thinking is provided to diagnose and monitoring Parkinson's.
Applicant states that the present invention illustrates class peptide of the invention and its preparation method and application by above-described embodiment,
But the invention is not limited in above-described embodiment, that is, do not mean that the present invention has to rely on above-described embodiment and could implement.It is affiliated
Those skilled in the art it will be clearly understood that any improvement in the present invention, equivalence replacement to raw material selected by the present invention and
The addition of auxiliary element, selection of concrete mode etc., within the scope of all falling within protection scope of the present invention and disclosing.
Claims (10)
1. a species peptide, it is characterised in that the class peptide includes following subunit：Ethylenediamine (I), piperonyl amine (II), β-the third
Propylhomoserin (III), naphthalidine (IV) and cysteine (V).
2. class peptide according to claim 1, it is characterised in that the sequence of the subunit of the class peptide is I-IV-III-II-
I-I-V。
3. class peptide according to claim 1 and 2, it is characterised in that the class peptide has following structure：
4. the preparation method of the class peptide according to any one of claim 1-3, it is characterised in that the preparation method passes through
Solid-phase synthesis synthesize；
Preferably, the preparation method is comprised the following steps：
(1) according to the subunit order of connection of class peptide, first subunit of the class peptide is connected on solid phase carrier；
(2) bromoacetic acid is reacted to be formed under the activation of activator with the amino of first subunit being connected on solid phase carrier
Amido link；
(3) product that second donor of subunit of class peptide is obtained with step (2) is reacted, is substituted off bromine atoms, it is complete
Into second connection of subunit；
(4) connection of bromoacetic acid and follow-up subunit is then repeated, until completing the connection of all subunits；
(5) the class peptide that obtains will be synthesized from solid phase carrier it will be cleaved and will obtain the class peptide.
5. preparation method according to claim 4, it is characterised in that step (1) described solid phase carrier is Rink amide
AM resins；
Preferably, step (1) is described is connected to solid phase carrier on solid phase carrier before first subunit of the class peptide
Carry out swelling；
It is when solid phase carrier is Rink amide AM resins, its is swelling, and cause Rink using hexahydropyridine deprotection
Amide AM resins expose amino.
6. the preparation method according to claim 4 or 5, it is characterised in that connect first subunit of the class peptide
During on to solid phase carrier, carried out in the presence of condensing agent and activator；
Preferably, the condensing agent is the tetramethylurea hexafluorophosphates of 2- (3'-N- oxos-BTA) -1,1', 3,3'-, O-
Any one in BTA-N, N, N', N'- tetramethylurea tetrafluoro boric acid or I-hydroxybenzotriazole or at least two
Combination；
Preferably, the activator is N- methylmorpholines；
Preferably, step (2) described activator is N, N'- DICs or dicyclohexylcarbodiimide；
Preferably, the temperature of step (2) described reaction is 20-40 DEG C；
Preferably, the time of step (2) described reaction is 20-100min；
Preferably, the temperature of step (3) described reaction is 20-40 DEG C；
Preferably, the time of step (3) described reaction is 50-150min；
Preferably, the decomposition agent for being used during step (5) described cracking includes the following composition of mass percent：95% trifluoro second
Acid, 2.5% ultra-pure water and 2.5% tri isopropyl silane；
Preferably, the group that the coupled reaction is will not participate in the connection procedure of the subunit of the polypeptide carries out group guarantor
Shield, then after the connection for completing all subunits, then carries out deprotection deprotection base.
7. a kind of amyloid detection agent, it is characterised in that the amyloid detection agent is included as in claim 1-3
Class peptide described in any one；
Preferably, the amyloid is alpha-synapse nucleoprotein.
8. a kind of pharmaceutical composition, it is characterised in that described pharmaceutical composition is included as any one of claim 1-3
Class peptide.
9. pharmaceutical composition according to claim 8, it is characterised in that described pharmaceutical composition is also comprising can pharmaceutically connect
The auxiliary material received；
Preferably, the pharmaceutically acceptable auxiliary material is in excipient, diluent, carrier, flavor enhancement, adhesive or filler
Any one or at least two combination.
10. prepared by the pharmaceutical composition described in the class peptide or claim 8 or 9 according to any one of claim 1-3
Application in the medicine of detection, diagnosis and/or the monitoring disease related to alpha-synapse nucleoprotein；
Preferably, the disease is Parkinson's.
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| CN111868072A (en) * | 2019-01-03 | 2020-10-30 | 京东方科技集团股份有限公司 | Peptoid compound, preparation method thereof, nano-carrier and pharmaceutical composition |
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| CN111868072B (en) * | 2019-01-03 | 2023-08-29 | 京东方科技集团股份有限公司 | Peptoid compound, preparation method thereof, nano carrier and pharmaceutical composition |
| CN113227117A (en) * | 2019-11-29 | 2021-08-06 | 京东方科技集团股份有限公司 | Peptoid compound and detection chip with peptoid compound coupled to surface |
| CN113227117B (en) * | 2019-11-29 | 2022-12-20 | 京东方科技集团股份有限公司 | Peptoid compound and detection chip with peptoid compound coupled to surface |
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