CN111366650A - Early prediction marker of endogenous small molecules of neurodegenerative diseases and application - Google Patents

Abstract

The invention discloses an endogenous small molecule early prediction marker for neurodegenerative diseases and application thereof, wherein the marker comprises 11, 12-epoxy-5, 8, 14-eicosatrienoic acid, morphine-D3, D-sphingosine, taurine, MAM2201-N- (5-chloropentyl) analogue, 5- (2,5-dihydroxyhexyl) oxo-2-ketone, oxalic acid, aniline, LPE22:1, 2,4-dinitrophenol, hippocampal alkali, N, N' -bis [4- (2,6-dimethylmorpholino) phenyl ] thiourea, erucic acid and arachidonic acid ethanol amide phosphate. The early prediction marker of the endogenous micromolecules of the neurodegenerative diseases has the characteristics of high accuracy and high sensitivity, and the prediction area AUC under the prediction line of some markers reaches over 0.95, so that the accuracy and the sensitivity of the early prediction marker are high, and the early prediction marker can be directly used for indexes of the neurodegenerative diseases.

Description

Early prediction marker of endogenous small molecules of neurodegenerative diseases and application

Technical Field

The invention belongs to the technical field of biomedicine, and particularly relates to an endogenous small molecule early prediction marker for neurodegenerative diseases and application thereof.

Background

Neurodegenerative diseases, including Alzheimer's Disease (AD) and Parkinson's Disease (PD), are related diseases in which the function of neurons deteriorates due to the interaction between aging and the environment of an individual. With the progress of aging in China and the world, the number of the neurodegenerative diseases increases year by year, and reaches 2019, the number of the neurodegenerative diseases exceeds 1500 ten thousand, which causes great burden to the country and families.

At present, the medicine which has no specific effect aiming at the neurodegenerative disease has irreversible pathogenesis progress, when a patient has obvious clinical phenotype, the disease course of the patient is usually in the middle and late stage, and the treatment can only slow down the development of the disease and can not radically reverse the damage of a cerebral nerve network. Therefore, the therapeutic principle for neurodegenerative diseases is early diagnosis and early treatment.

Unfortunately, there is no effective technical means for early diagnosis of neurodegenerative diseases. By the time the patient develops obvious symptoms, the symptoms often reach the middle and late stage of the disease and are irreversible, so that the development of a new early diagnosis marker of neurodegenerative diseases is urgently needed.

Although the pathogenesis of neurodegenerative diseases is different, and the early clinical manifestations are different, the degeneration of the function of brain neurons is the main pathogenesis, so essentially the neurodegenerative diseases are related to brain aging. Since all neurodegenerative diseases have the same characteristics of aging and degeneration of cerebral neurons, it is possible to develop common biomarkers of neurodegenerative diseases, and since the aging problem is severe in China at present, among which the elderly aged 65 years or older will increase from 1.19 hundred million in 2010 to 3.6 hundred million in 2050 (accounting for 25.6% of the total population), and the risk of suffering from various neurodegenerative diseases increases year by year, it is necessary to develop completely new biomarkers of neurodegenerative diseases.

Disclosure of Invention

In view of the above, the present invention provides an early prediction marker of endogenous small molecules of neurodegenerative diseases and applications thereof.

In order to solve the technical problems, the invention discloses an endogenous small molecule early prediction marker for neurodegenerative diseases, which comprises one or more of 11, 12-epoxy-5, 8, 14-eicosatrienoic acid, morphine-D3, D-sphingosine, taurine, MAM2201-N- (5-chloropentyl) analogue, 5- (2,5-dihydroxyhexyl) oxo-2-one, oxalic acid, aniline, LPE22:1, 2,4-dinitrophenol, hippocampal, N, N' -bis [4- (2,6-dimethylmorpholino) phenyl ] thiourea, erucic acid and arachidonic acid ethanol amide phosphate.

The invention also discloses application of the endogenous small molecule early prediction marker for the neurodegenerative disease in preparing a neurodegenerative disease prediction product, wherein the prediction product comprises a reagent for detecting the one or more markers, and the reagent is used for judging whether a subject has the neurodegenerative disease by detecting the concentration of the one or more markers in the body of the subject.

Optionally, in particular the concentration of one or more markers in the blood of the subject is detected.

Alternatively, the markers 11, 12-epoxy-5, 8, 14-eicosatrienoic acid, morphine-D3, D-sphingosine, taurine, MAM2201-N- (5-chloropentyl) analog, 5- (2,5-dihydroxyhexyl) oxo-2-one are present in significantly higher concentrations in patients with neurodegenerative diseases than in patients with non-neurodegenerative diseases.

Alternatively, the markers oxalic acid, aniline, LPE22:1, 2,4-dinitrophenol, hippocampal base, N' -bis [4- (2,6-dimethylmorpholino) phenyl ] thiourea, erucic acid, arachidonic acid ethanolamide phosphate are present in significantly lower concentrations in patients with neurodegenerative diseases than in patients with non-neurodegenerative diseases.

The invention also discloses a composition for early prediction of neurodegenerative diseases, which comprises one or more than one of the endogenous small molecule early prediction markers of the neurodegenerative diseases.

Compared with the prior art, the invention can obtain the following technical effects:

the early prediction marker of the endogenous micromolecules of the neurodegenerative diseases has the characteristics of high accuracy and high sensitivity, and the prediction area AUC under the prediction line of some markers reaches over 0.95, so that the accuracy and the sensitivity of the early prediction marker are high, and the early prediction marker can be directly used for indexes of the neurodegenerative diseases.

Of course, it is not necessary for any one product in which the invention is practiced to achieve all of the above-described technical effects simultaneously.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention and not to limit the invention. In the drawings:

FIG. 1 is a ROC graph and corresponding molecular formula of 11, 12-epoxy-5, 8, 14-eicosatrienoic acid as a neurodegenerative marker of the present invention for predicting neurodegenerative diseases; wherein, A is ROC curve diagram of 11, 12-epoxy-5, 8, 14-eicosatrienoic acid, and B is molecular formula of 11, 12-epoxy-5, 8, 14-eicosatrienoic acid;

FIG. 2 is a ROC graph of the neurodegenerative marker morphine-D3 of the present invention predicting neurodegenerative disease and the corresponding molecular formula; wherein, A is ROC curve chart of morphine-D3, B is molecular formula of morphine-D3;

FIG. 3 is a ROC plot and corresponding molecular formula of the neurodegenerative marker D-sphingosine of the present invention for predicting neurodegenerative disease; wherein A is ROC curve diagram of D-sphingosine, and B is molecular formula of D-sphingosine;

FIG. 4 is a ROC graph of taurine as a neurodegenerative marker of the present invention predicting neurodegenerative disease and the corresponding molecular formula; wherein, A is an ROC curve chart of taurine, and B is a molecular formula of taurine;

FIG. 5 is a ROC plot and corresponding molecular formula of the neurodegenerative marker MAM2201-N- (5-chloropentyl) analog of the present invention for predicting neurodegenerative disease; wherein, A is the ROC curve diagram of the MAM2201-N- (5-chloropentyl) analogue, and B is the molecular formula of the MAM2201-N- (5-chloropentyl) analogue;

FIG. 6 is a ROC plot and corresponding molecular formula for the neurodegenerative marker 5- (2,5-dihydroxyhexyl) oxo-2-one of the present invention predicting neurodegenerative disease; wherein A is ROC curve of 5- (2,5-dihydroxyhexyl) oxo-2-one, and B is molecular formula of 5- (2,5-dihydroxyhexyl) oxo-2-one;

FIG. 7 is a ROC plot and corresponding molecular formula for oxalic acid, a neurodegenerative marker of the present invention, to predict neurodegenerative disease; wherein A is an ROC curve chart of oxalic acid, and B is a molecular formula of oxalic acid;

FIG. 8 is a ROC plot and corresponding molecular formula for the neurodegenerative marker aniline of the present invention to predict neurodegenerative; wherein, A is ROC curve diagram of aniline, B is molecular formula of aniline;

FIG. 9 is a ROC graph of the neurodegenerative marker LPE22:1 of the present invention predicting neurodegenerative disease and the corresponding molecular formula; wherein, A is an ROC curve chart of LPE22:1, and B is a molecular formula of LPE22: 1;

FIG. 10 is a ROC graph of the neurodegenerative marker 2,4-dinitrophenol of the present invention predicting neurodegenerative disease and the corresponding molecular formula; wherein A is ROC curve diagram of 2,4-dinitrophenol, B is molecular formula of 2, 4-dinitrophenol;

FIG. 11 is a ROC plot and corresponding molecular formula of hippocampal base, a neurodegenerative marker of the present invention, for predicting neurodegenerative disease; wherein, A is ROC curve chart of hippocampus alkaloid, B is molecular formula of hippocampus alkaloid;

FIG. 12 is a ROC graph of the neurodegenerative marker N, N' -bis [4- (2,6-dimethylmorpholino) phenyl ] thiourea of the present invention for predicting neurodegenerative disease and the corresponding molecular formula; wherein, A is ROC curve diagram of N, N '-bis [4- (2,6-dimethylmorpholino) phenyl ] thiourea, and B is molecular formula of N, N' -bis [4- (2,6-dimethylmorpholino) phenyl ] thiourea;

FIG. 13 is a ROC plot and corresponding molecular formula for the neurodegenerative marker erucic acid of the present invention predicting neurodegenerative; wherein, A is an ROC curve diagram of erucic acid, and B is a molecular formula of the erucic acid;

FIG. 14 is a ROC plot and corresponding molecular formula of arachidonic acid ethanolamide phosphate as a neurodegenerative marker of the present invention predicting neurodegenerative diseases; wherein A is ROC curve diagram of arachidonic acid ethanol amide phosphate, and B is molecular formula of arachidonic acid ethanol amide phosphate.

Detailed Description

The following embodiments are described in detail with reference to the accompanying drawings, so that how to implement the technical features of the present invention to solve the technical problems and achieve the technical effects can be fully understood and implemented.

Example 1 endogenous Small molecule early predictive markers for neurodegenerative diseases

The invention discloses an endogenous small molecule early prediction marker for neurodegenerative diseases, which can be used for early prediction of the neurodegenerative diseases by single or combination of two or more than two of 11, 12-Epoxy-5, 8, 14-eicosatrienoic acid (11,12-Epoxy- (5Z,8Z,11Z) -icosaterioic acid), Morphine-D3 (Morphine-D3), D-Sphingosine (D-Sphingosine), Taurine (Taurine), MAM2201-N- (5-chloropentyl) analogue (MAM2201-N- (5-chloropentanyl) analogue), 5- (2,5-dihydroxyhexyl) oxo-2-one (5- (2, 5-dihydroxyyl) oxolan-2-one), 3,4, 5-trihydroxy-1-cyclohexene carboxylic acid (kishinoic acid), aniline (Aniline), lysophosphatidylethanolamine LPE22: one or more of 1, 2,4-Dinitrophenol (2,4-Dinitrophenol), hippocampal base (Harmaline), N '-bis [4- (2,6-dimethylmorpholino) phenyl ] thiourea (N, N' -di [4- (2,6-dimethylmorpholino) phenyl ] thiourea), Erucic acid (Erucic acid), arachidonic acid ethanolamide phosphate (Arachidonoylethanolamide phosphate), and their corresponding smile-format structures are shown in table 1.

TABLE 1 molecular formula Smile Structure of each marker

Of the 14 neurodegenerative disease markers, the significant increase in neurodegenerative disease patients compared to non-neurodegenerative disease patients was: 6 of 11, 12-epoxy-5, 8, 14-eicosatrienoic acid, morphine-D3, D-sphingosine, taurine, MAM2201-N- (5-chloropentyl) analogue, 5- (2,5-dihydroxyhexyl) oxo-2-one.

Of the 14 neurodegenerative disease markers, the significant decrease in neurodegenerative disease patients compared to non-neurodegenerative disease patients was: 3,4, 5-trihydroxy-1-cyclohexene carboxylic acid, aniline, lysophosphatidylethanolamine LPE22:1, 2,4-dinitrophenol, hippocampal alkali, N, N' -bis [4- (2,6-dimethylmorpholino) phenyl ] thiourea, erucic acid, arachidonic acid ethanol amide phosphate ester totally 8.

Through prospective research on 121 blood samples of non-neurodegenerative disease patients and 32 blood samples of neurodegenerative disease patients in China (wherein 14 blood samples of Alzheimer patients and 18 blood samples of neurodegenerative disease patients), the change of each marker in neurodegenerative disease and the corresponding area AUC under a prediction line are shown in Table 2, and ROC curves of the 14 markers for predicting neurodegenerative disease are shown in FIGS. 1A, 2A and 3A-14A; the molecular structures of the 14 blood neurodegenerative disease diagnosis markers are shown in figures 1B, 2B, 3B-14B.

As can be seen from table 2 and the accompanying figures, AUC characterizes the predictive power of the marker, and AUC ═ 1 indicates 100% accuracy and sensitivity. The AUC of the 14 corresponding markers for distinguishing healthy people reaches over 0.90, and the area under the line of some markers such as morphine-D3 and arachidonic acid ethanol amide phosphate reaches over 0.95, which shows that the markers have higher accuracy and sensitivity in predicting neurodegenerative diseases.

Table 2 change in each marker in neurodegenerative patients and corresponding area under the predictive line ROC

Example 2 diagnosis of neurodegenerative diseases with 11, 12-Epoxy-5, 8, 14-eicosatrienoic acid (11,12-Epoxy- (5Z,8Z,11Z) -icosaterienoic acid) as single marker:

the specific implementation method comprises the following steps:

1. obtaining a sample of a subject, and carrying out pretreatment: collecting blood sample of a subject according to a standard and reasonable operation process, adding 100 mu L of blood into 350 mu L of methanol, performing vortex mixing, performing high-speed centrifugation, taking supernatant, performing concentration drying, adding 100 mu L of acetonitrile for redissolution, and performing UHPLC-QTOFMS test directly through a filter plug.

2. Data processing: and after the UHPLC-QTOFMS is tested, collecting UHPLC-QTOFMS data for analysis. The data processing uses XCMS program, and the parameter setting refers to software description. After the XCMS program treatment, charge-to-mass ratios, retention times, peak areas and secondary mass spectrum indexes of all metabolites are obtained.

3. Determination of the concentration of 11, 12-epoxy-5, 8, 14-eicosatrienoic acid: mass-to-charge-mass ratios and mass spectrum secondary fragments are determined by aligning secondary mass spectrum databases. The concentration of 11, 12-epoxy-5, 8, 14-eicosatrienoic acid in blood was determined by the ratio of its peak area to the peak area of the standard, wherein metabolite concentration determination was made by the Baiqu biomedical science and technology Limited in Shanghai.

4. Diagnosis of neurodegenerative diseases: the concentration of 11, 12-epoxy-5, 8, 14-eicosatrienoic acid in the blood of the subject measured as described above is compared with a threshold value for patients with non-neurodegenerative diseases. If the value is above the threshold for a non-neurodegenerative patient, then the subject has a higher likelihood of neurodegenerative disease. Wherein the concentration of 1, 12-epoxy-5, 8, 14-eicosatrienoic acid in the blood of non-neurodegenerative disease patients is less than 30 nmol/L. The results showed 0.9765 for the area under the characteristic curve. The marker can be accurately applied to the diagnosis of neurodegenerative diseases.

Taking a sample as an example, the concentration of 11, 12-epoxy-5, 8, 14-eicosatrienoic acid in the sample blood is 678nmol/L, which is higher than the threshold value (90nmol/L) for the general population, indicating that the individual has a higher probability of neurodegenerative disease.

Example 3 diagnosis of neurodegenerative diseases with the marker D-sphingosine as single marker:

the D-sphingosine concentration of the subject was determined in the same manner as in 1-3 steps of example 2. Comparing the measured concentration of D-sphingosine in the blood of the subject with a threshold value for a patient with a non-neurodegenerative disease. If the value is above the threshold for a non-neurodegenerative patient, then the subject has a higher incidence of neurodegenerative disease. The result shows that the area under the characteristic curve line is 0.9161, which proves that the marker can be more accurately applied to the diagnosis of the neurodegenerative diseases.

In the case of a sample, the concentration of D-sphingosine in the blood of the sample is 171nmol/L, which is higher than the threshold value for the compound in patients with non-neurodegenerative diseases (55nmol/L), indicating that the individual has a higher probability of suffering from neurodegenerative diseases.

Example 4

The diagnosis is carried out by combining two metabolites, namely 11, 12-epoxy-5, 8, 14-eicosatrienoic acid and D-sphingosine, and taking a sample as an example, the concentration of 11, 12-epoxy-5, 8, 14-eicosatrienoic acid in the sample is 821nmol/L (which is greater than the normal threshold value of 90nmol/L) and the concentration of sphingosine in the sample is 263nmol/L (which is greater than the normal threshold value of 55nmol/L), and the two metabolites are elevated in patients with neurodegenerative diseases by looking up a table 2. The above two metabolite combination data indicate that the subject is most likely to suffer from neurodegenerative diseases and should respond early.

While the foregoing description shows and describes several preferred embodiments of the invention, it is to be understood, as noted above, that the invention is not limited to the forms disclosed herein, but is not to be construed as excluding other embodiments and is capable of use in various other combinations, modifications, and environments and is capable of changes within the scope of the inventive concept as expressed herein, commensurate with the above teachings, or the skill or knowledge of the relevant art. And that modifications and variations may be effected by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.

Claims (6)

1. An endogenous small molecule early prediction marker for neurodegenerative diseases is characterized by comprising one or more of 11, 12-epoxy-5, 8, 14-eicosatrienoic acid, morphine-D3, D-sphingosine, taurine, MAM2201-N- (5-chloropentyl) analogue, 5- (2,5-dihydroxyhexyl) oxo-2-ketone, oxalic acid, aniline, LPE22:1, 2,4-dinitrophenol, hippocampal alkali, N, N' -bis [4- (2,6-dimethylmorpholino) phenyl ] thiourea, erucic acid and arachidonic acid ethanol amide phosphate.

2. Use of the endogenous small molecule early predictive marker of a neurodegenerative disease as claimed in claim 1 for the preparation of a product for predicting a neurodegenerative disease, wherein the product comprises a reagent for detecting the one or more markers, wherein the reagent determines whether the subject suffers from a neurodegenerative disease by detecting the concentration of the one or more markers in the subject.

3. Use according to claim 2, in particular to detect the concentration of one or more markers in the blood of a subject.

4. The use of claim 2, wherein the marker is 11, 12-epoxy-5, 8, 14-eicosatrienoic acid, morphine-D3, D-sphingosine, taurine, MAM2201-N- (5-chloropentyl) analog, 5- (2,5-dihydroxyhexyl) oxo-2-one, at a significantly higher concentration in patients with neurodegenerative diseases than in patients with non-neurodegenerative diseases.

5. The use according to claim 2, wherein the markers oxalic acid, aniline, LPE22:1, 2,4-dinitrophenol, hippocampal base, N' -bis [4- (2,6-dimethylmorpholino) phenyl ] thiourea, erucic acid, arachidonic acid ethanolamide phosphate are present in significantly lower concentrations in patients with neurodegenerative diseases than in patients with non-neurodegenerative diseases.

6. An early stage prediction composition for neurodegenerative diseases, which comprises one or more endogenous small molecule early stage prediction markers for neurodegenerative diseases as claimed in claim 1.

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