CN108498798A - It is a kind of prevention skeletal muscle atrophy medicine target spot and its application - Google Patents
It is a kind of prevention skeletal muscle atrophy medicine target spot and its application Download PDFInfo
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Abstract
The present invention relates to a kind of medicine target spot of prevention skeletal muscle atrophy and its applications.Application the present invention provides branch α ketoacid dehydrogenase complexes (BCKDC) or branch α ketoacid dehydrogenases kinases (BCKDK) as drug target in preventing or treating skeletal muscle atrophy, and BCKDC and BCKDK is preparing the application in preventing or treating skeletal muscle atrophy drug, the method that specially anti-skeletal muscle atrophy drug is screened in inside and outside：Using BCKDC or its gene or BCKDK or its gene as drug effect object, the inhibitor of BCKDC or the activator of BCKDK are picked out as the candidate primary dcreening operation drug for preventing or treating skeletal muscle atrophy.The present invention is from its critical limitation metabolic enzyme regulatory mechanism of tumor cachexia metabolic regulation angle analysis, and pharmaceutical intervention target spot is determined, it has potential clinical value in skeletal muscle atrophy treatment, and new platform is provided for the screening of skeletal muscle atrophy medicine.
Description
Technical field
The invention belongs to biotechnologies, and in particular to it is a kind of prevention skeletal muscle atrophy medicine target spot and its answer
With.
Background technology
Skeletal muscle atrophy be common in cachectic patients (including malignant tumour, chronic heart failure, chronic obstructive pulmonary disease,
The cachexia that the diseases such as chronic kidney disease, rheumatoid arthritis induce), gerontal patient and muscular dystrophy patient.It shows as
Protein catabolism increases, and anabolism is insufficient, and severe weight is caused to decline, and muscle is lost.The generation of skeletal muscle atrophy is not
Patients ' life quality is only reduced, shortens patient survival, and seriously affect the implementation of patient base's disease treatment scheme, is reduced
Drug therapy sensibility, increases complication.
Skeletal muscle atrophy pathogenesis is mainly chronic inflammation and oxidative stress causes insulin resistance and catabolism to add
Speed, but anti-inflammatory and inhibition oxidative stress is limited to the preventive and therapeutic effect of skeletal muscle atrophy, improves insulin resistance and nutrition treatment
Symptom cannot be improved and extend patient survival.Therefore, the pathogenesis of new breakthrough is found based on understanding and therapy target is gone forward side by side
Row effective prevention has great importance for the prevention of skeletal muscle atrophy.
BCAA (branched-chain amino acid, Branched-Chain Amino Acid) is essential amino acid, including figured silk fabrics ammonia
Acid, leucine and isoleucine are not only the important composition ingredient of myogen, and are of great significance to protein synthesis.
Clinical research and results of animal show that the supplement of BCAA can increase cachectic patients' blood plasma prealbumin, hemoglobin and blood
It is horizontal to starch transferrin, increases gastrocnemius and tibialis anterior quality, reduces E3 ligase expressions.BCKDC (branched-chain alpha-ketoacids
Dehydrogenase complex, Branched-Chain α-Keto acid Dehydrogenase Complex) it is Intramitochondrial multienzyme
Compound participates in BCAA metabolism, is its restricted irreversible enzyme.By BCKD kinases, (branched-chain alpha-keto acid dehydrogenase swashs BCKDC activity
Enzyme, Branched-Chain α-Keto acid Dehydrogenase Kinase) negative regulation.It is existing that researches show that 2 types sugar
Urine is sick, Metabolic syndrome is sought peace, and obese patient shows BCKDC expression and activity exception, but mutation and the dysfunction of BCKDC
The 2-ketoacid concentration that serum BCAA and its metabolism are generated can be caused to increase, cause central nervous system dysfunction, or even experiment
Animal dead.BCKDK is focused primarily upon currently based on the study on regulation of BCKDC, literature survey finds BCKDK inhibitor BT2 mesh
The preceding positive treatment explored for diseases such as insulin resistances；And there is document to point out to carry out proliferative diseases using BCKDK inhibitor
Treatment or prevention method, as international monopoly WO 2017/155467 disclose it is a kind of treat or prevent proliferative diseases side
Method, by being selected from the accumulation of branched-chain amino acid (BCAA), the inhibition of the activity of BCAA catabolic enzymes or transcript level, drop
Low fatty acyl carnitine level (C5：1) at least one is characterized and/or is diagnosed, and this method includes giving the enhancing of BCAA catabolism
Agent and/or branched-chain alpha-ketoacid dehydrogenase complex (BCKDC) kinase inhibitor.
But there is no disclose being associated between BCKDC or BCKDK and skeletal muscle atrophy for the prior art.
Invention content
The present invention is directed to the treatment of skeletal muscle atrophy, and in the course of the research, inventor has found BCKDC or BCKDK and bone
There is close relevance, and it is thus found that a kind of medicine target spot of new prevention skeletal muscle atrophy between amyotrophia,
Additionally provide a kind of screening technique of skeletal muscle atrophy prevention or medicine based on the drug target.
To achieve the above object, the present invention adopts the following technical scheme that：
The present invention provides branched-chain alpha-ketoacid dehydrogenase complex (BCKDC) or branched-chain alpha-keto acid dehydrogenase kinases (BCKDK)
As application of the drug target in preventing or treating skeletal muscle atrophy.
The present invention also provides branched-chain alpha-ketoacid dehydrogenase complex (BCKDC) or branched-chain alpha-keto acid dehydrogenase kinases
(BCKDK) application in preventing or treating skeletal muscle atrophy drug is being prepared.
In order to advanced optimize above-mentioned technical proposal, the technical measures that the present invention takes further include：
Further, the skeletal muscle atrophy includes Sarcopenia, amyotrophia, muscular dystrophy, cachexia, wherein the evil
Sick matter includes at least in malignant tumour, chronic heart failure, chronic obstructive pulmonary disease, chronic kidney disease, rheumatoid arthritis
The cachexia of one or more inductions.
Further, the BCKDC or BCKDK includes internal preparing the application in preventing or treating skeletal muscle atrophy drug
Outer screening prevents or the drug for the treatment of skeletal muscle atrophy comprising following steps：With BCKDC or its gene or BCKDK or its base
Because as drug effect object, picking out the inhibitor of BCKDC or the activator of BCKDK as prevention or treatment skeletal muscle atrophy
Candidate primary dcreening operation drug.
Further, the inside and outside screening prevents or includes the step for the treatment of the drug of skeletal muscle atrophy：With BCKDC or
The gene of BCKDC picks out the inhibitor of BCKDC as the candidate for preventing or treating skeletal muscle atrophy as drug effect object
Primary dcreening operation drug；And using the gene of BCKDK or BCKDK as drug effect object, pick out the activator of BCKDK as preventing or
Treat the candidate primary dcreening operation drug of skeletal muscle atrophy.
Further, the inhibitor is the compound for having inhibition to BCKDC；Further, the inhibition effect
Fruit includes inhibiting BCKDC activity, or can inhibit the genetic transcription or expression of BCKDC.
Further, the activator is the compound for having activation effect to BCKDK；Further, the activation effect
Fruit includes inhibiting BCKDC activity, or can promote the genetic transcription or expression of BCKDK.
Further, it includes enzyme kinetics method to judge whether candidate primary dcreening operation drug can inhibit the active methods of BCKDC.Its
Any suitable other methods in the prior art can be used.
Further, judge whether candidate primary dcreening operation drug can inhibit genetic transcription and the base of expression or promotion BCKDK of BCKDC
Because the method transcribed and expressed includes the following steps：Cell or the mouse that normal expression BCKDC or BCKDK are provided, in medicine to be measured
Object carries and cultivates the cell or mouse in the presence of the carrier of drug to be measured, detect BCKDC or BCKDK transcription and
Whether expression changes.Any suitable other methods in the prior art can also be used in it.
Further, one kind during the activator of the inhibitor of the BCKDC and the BCKDK at least have the effect that or
It is a variety of：Inhibit skeletal muscle tissue E3 ubiquitination ligases MuRF1 and MAFbx/Atrogin-1 expression；Inhibit skeletal muscle tissue
Ubiquitin protein enzyme body activates；Regulate and control expression and the phosphorylation of the FoxO1 and FoxO3 of skeletal muscle tissue；Promote skeletal muscle tissue
MTOR expression increases or phosphorylation degree increases；Promote skeletal muscle tissue MyHC, Myf4, Myogeinin, Myf5, MyoD and
Pax3 expression increases；Increase skeletal muscle mass or weight；Increase hand grip endurance.
Compared with prior art, the invention has the advantages that：
(1) skeletal muscle atrophy is tumor cachexia important feature, and the present invention is using cachexia skeletal muscle atrophy as model, from evil
The branched-amino acid metabolic of sick matter changes feature and sets out, and explores the clinical meaning and regulatory mechanism of critical limitation sexual factor BCKDC,
It analyzes it and regulates and controls the effect that kinase b CKDK synthesizes protein level of skeletal muscle and degrades.
(2) it is based on inventor project team metabolic enzyme research experience for many years, with13Leucine is analysed in C flag indication difference group credit
The qualitative and quantitative variation of catabolic intermediate, with liquid chromatography-mass spectrometry to critical limitation sexual factor BCKDC
Expression and activity are measured, with gene overexpression/gene knockout means from inside and outside experimental analysis BCKDK to branched-chain amino acid
The regulating and controlling effect and mechanism of metabolism and skeletal muscle atrophy.
(3) mechanism based on tumor cachexia skeletal muscle atrophy explores BCKDK and mediates BCKDC dysfunctions to skeletal muscle
The effect of atrophy, is tested by inside and outside, horizontal from molecule, cell and tissue, and research BCKDK interference/overexpression is to BCKDC tables
Up to function controlling mechanism, and to the effect that branched-amino acid metabolic and protein level of skeletal muscle synthesize, prevented for skeletal muscle atrophy
Novel targets are provided.
The present invention is dry from its critical limitation metabolic enzyme regulatory mechanism of tumor cachexia metabolic regulation angle analysis and drug
Pre- target spot has potential clinical value in skeletal muscle atrophy treatment.
Description of the drawings
Fig. 1 shows the foundation of tumor cachexia diagnostic equation and verifications：Before random forest analysis discloses differentiation cachexia
Phase, the cachexia phase, the non-cachexia of tumour and normal healthy controls 15 most important metabolins (A).It concentrates in verification and is analyzed with ROC
Sensitivity and specificity (B) of the diagnosis marker to sample packet.
Fig. 2 indicates that skeletal muscle tissue dynamic metabolin changes and significantly change the impact factor of metabolic pathway.
Fig. 3 indicates that high resolution NMR Spectrum Analysis (NMR) shows dexamethasone and tumor cachexia group sample α -one
Dissident's acid signal (chemical shift [ppm] 51.6 and 26.9) reduces, and α-ketoisocaproic acid concentration is prompted to decline.
Fig. 4 indicates to handle the C2C12 myotubules of differentiation to BCKDK/BCKDC signals with dexamethasone, Formoterol and BT2
Pathway protein and myogen constituent Myogeinin, MyHC expression influence.
Fig. 5 indicates that immunocyte dyeing shows that induced by dexamethasone amyotrophia, BT2 also can induce amyotrophia, and ground plug rice
When pine and BT2 are handled simultaneously, amyotrophia is even more serious.Blue is DAPI nuclear targetings, and red is MyHC dyeing.
It is micro- after Fig. 6 indicates the C2C12 multinuclears myotubule of differentiation with the adenovirus processing 48h of 3 kinds of interference BCKDK expression
Under the microscope, green fluorescence shows the expression of BCKDK, and 3 kinds of virus treateds cause muscle fiber atrophy, wherein with third
Adenovirus (BCKDK-V3) effect of kind structure is most apparent (A)；It then carries out immunocyte dyeing and shows MyHC expressions, it can
See that 3 kinds of virus treateds cause muscle fibre MyHC expression to reduce, wherein most apparent (B) with BCKDK-V3 effects.
Fig. 7 indicate 3 kinds interference BCKDK expression adenovirus processing differentiation C2C12 multinuclears myotubule to BCKDK,
The mRNA expression of BCKDP, BCKDE1A, MyoD, Myogeinin, MyHC, MuRF1 and MAFbx influence.(*) is indicated and control group
Virus is compared, and has notable significant difference (p<0.05).
Fig. 8 shows interference BCKDK expression adenovirus processing differentiation C2C12 multinuclears myotubule to BCKDK, BCKDE1A,
The protein expression of MyoD, Myogeinin and MyHC influence.
Fig. 9 indicate the adenovirus of gastrocnemius local injection interference BCKDK expression to gastrocnemius and tibialis anterior BCKDK,
The mRNA expression influences (A) of BCKDP, the protein expression influence (B) of gastrocnemius MyoD and MuRF1 and gastrocnemius MyoD,
The mRNA expression of Myogeinin and MyHC influences (C).(*) is indicated compared with control group virus, is had notable significant difference (p
<0.05)。
Specific implementation mode
The present invention provides branched-chain alpha-ketoacid dehydrogenase complexes or branched-chain alpha-keto acid dehydrogenase kinases as drug effect
Application and BCKDC or BCKDK of the target spot in preventing or treating skeletal muscle atrophy are preparing prevention or treatment skeletal muscle atrophy
Application in drug.
With reference to the accompanying drawings and examples, the specific implementation mode of the present invention is further described.Following embodiment is only
For clearly illustrating technical scheme of the present invention, and not intended to limit the protection scope of the present invention.
Embodiment 1
The present embodiment is the metabolism change for detecting tumor patient, and experimental method is as follows：
(1) 222 tumor patients (including 84 tumor cachexia patients, 33 cachexia early stage patients and 105 are collected
The tumor patient of body mass stable) and 74 normal healthy controls crowds clinical data and serum/urine specimen, patient be divided into test set
Diagnostic model structure is carried out after collecting with verification.
(2) whole blood collects in the EP pipes without containing anti-coagulants, and the light yellow serum in upper layer is collected in centrifugation.
(3) it takes 200 μ l serum that EP pipes are added, adds 400 μ l and contain D2O and NaN3PBS buffer solution, shake mixing, centrifugation
Impurity in serum is removed, small molecule metabolites are only retained.
(4) Aspirate supernatant is transferred in the NMR pipes of 5mm after centrifuging, and is measured.
Experimental result is shown in Fig. 1, metabonomic analysis has apparent separation, wherein branched-amino acid metabolic different between showing 4 groups
It is often the most important metabolic characteristics of tumor cachexia.
The present embodiment is analyzed using NMR, and advantage is：As a kind of nondestructive analysis method, NMR analyses do not destroy
Sample structure and information need not also carry out complicated sample pretreatment, therefore have good reproducibility.In addition to this, NMR
The metabolism group investigative technique on basis measures1H、13C or31P, as long as selecting the suitable train of impulses, so that it may to realize to a certain
The scanning of atom, you can prejudice-free to tell whole otherness metabolite signals.It is overlapped for having in complex samples
The identification of the metabolome of signal, two dimensional NMR techniques can provide the atom composition and atom connection type information of molecule, directly really
Determine metabolite structures, this ensures that NMR can disclose the metabolic alterations situation of airframe systems from qualitative and quantitative angle.
Embodiment 2
The present embodiment is the metabolism change for detecting tumor cachexia animal model, and experimental method is as follows：
(1) tumor cachexia animal model is established：When B16 black cancer cells were passaged to for 4 generation, are cleaned and made with PBS
At 1 × 107The suspension of/ml density, is inoculated in that male C 57 BL/6 J mouse right fore back side back upper place is subcutaneous, and every mouse connects
0.1~0.2ml of kind.Inoculation puts to death mouse after about 3 weeks, takes tumor mass to pass on, stable tumor cachexia model is obtained after the 3rd generation.
(2) experiment mice is in being inoculated with the 11st day visible apparent tumor mass, and the next day subsequent or the daily tumor mass that measures is long and wide, meter
Calculate knurl weight.
(3) the 21st days or so, blood is taken, while putting to death animal and taking out the heart, liver, spleen, lung, kidney, epididymal adipose tissues, gastrocnemius and shin
Flesh before bone, weighs, and being transferred to ultra low temperature freezer after quick-frozen in liquid nitrogen freezes.
(4) high-resolution Magic angle spinning spectroscopic technique is used to carry out gastrocnemius metabolite analysis, with CPMG Spectrum Analysis blood
Clear metabolin.
Experimental result is shown in Fig. 2, change as a result, it has been found that 43 kinds of metabolites in 13 kinds of metabolites and serum of gastrocnemius have
Become, metabolic pathway analysis is integrated based on metabolin, finds 5 species specific metabolic characteristics, including hypoglycemia, high ketoboidies, neutrality
Amino acid increases, intermediate metabolites is disorderly and branched-amino acid concentration reduces.
The present embodiment has been narrowed compound information peak using high-resolution Magic angle spinning spectroscopic technique, so as to provide satisfaction
The high-resolution metabolin spectrogram that metabonomic analysis requires is directly realized by Solid State Structure detection and analysis, discloses histopathology shape
The metabolite profile of state.This test sera small molecule metabolites are measured using the CPMG trains of impulses, can be in the knot for not destroying sample
In the case of structure and property, the influence of lipid macromolecular metabolite signals is shielded, in real time and dynamic without being biased to detects all small point
Sub- metabolite signals.
Embodiment 3
The present embodiment is being associated in vitro study BCKDC and myotube atrophy, and experimental method is as follows：
(1) DMEM high glucose medium of the purchase without leucine is added 52.5mg's in the 500ml culture mediums
Leucine-13C6 (CLM-2262-H-PK, Cambridge Isotope Laboratories, Inc., Andover, MA), training
The multinuclear myotubule of nutrient.
(2) cell is grouped, and dexamethasone and tumor cachexia patients serum handle C2C12 multinuclear myotubules, cultivate 48h.
(3) culture medium supernatant cell pyrolysis liquid is collected, the solvent of containing the internal standard is added after sample concentration freeze-drying, it is total with nuclear-magnetism
It shakes or mass spectrum carries out leucine intermediate metabolites qualitative and quantitative analysis, and compare metabolin change level, analyze metabolin
Upstream and downstream protease expression quantity.
(4) experimental result is shown in Fig. 3, dexamethasone and tumor cachexia patients serums to handle C2C12 multinuclear myotubules, equal energy
It is that isovaleryl is auxiliary through BCKDC metabolic conversions to cause the reduction of α-ketoisocaproic acid (α-Ketoisocaproate) signal, α-ketoisocaproic acid
Enzyme A.The results show that dexamethasone and tumor cachexia patients serum induce adjoint BCDKC increased activities during amyotrophia.
The present embodiment passes through13C flag otherness metabolin carries out disease induction and medicine stimulation, with nuclear magnetic resonance
With the qualitative and quantitative variation of Mass Spectrometer Method metabolin, not only it can be found that the specific metabolic under myotube atrophing state changes feelings
Condition and drug target, and the disadvantages such as traditional metabolism group detection technique sensitivity can be overcome low.
Embodiment 4
The present embodiment is influence of the in vitro study BCKDK inhibitor BT2 processing to myotube atrophy, and experimental method is as follows：
(1) C2C12 sarcoblasts are seeded to six orifice plates, are trained in containing 10% new fetal calf serum DMEM high glucose mediums
After supporting 2~4d, observe cell density, wait for cell density about 80%, replace 2% horse serum DMEM high glucose medium culture 4d with
On, induction C2C12 myoblast differentiations are at multinuclear myotubule.
(2) cell is grouped, and BCKDK inhibitor BT2 handles C2C12 multinuclear myotubules, while DEX model groups and control is arranged
Group cultivates 48h.
(3) cell is collected, cell pyrolysis liquid is prepared, Western Blot detect muscle fibre different differentiation phases biological marker
The protein expression situation of object Myogeinin, MyHC, and the influence to BCKDK, BCKDE1A and phosphorylation BCKDE1A albumen.
Experimental result is shown in Fig. 4, BCKDK inhibitor BT2 processing causes BCKDK expression to decline, BCKDE1A phosphorylation degree liters
Height, myogen constitutive character ingredient Myogeinin and MyHC expression decline.Immunocyte dyeing shows BCKDK inhibitor BT2 processing
Muscle fiber atrophy, myogen MyHC expression is caused to significantly reduce, the result is shown in Fig. 5.
Embodiment 5
The present embodiment is that in vitro study interferes BCKDK expression to influence muscle fibre differentiation and the synthesis of multinuclear myotubule albumen,
Its experimental method is as follows：
(1) induction C2C12 myoblast differentiations are at multinuclear myotubule.
(2) cell is grouped, the C2C12 multinuclear myotubules of the adenovirus processing differentiation of 3 kinds of interference BCKDK expression, culture 48
Hour.
The adenovirus information of (3) 3 kinds of interference BCKDK expression is shown in Table 1.
Table 1：Interfere the essential information of BCKDK expression adenovirus
(4) MyHC differential expressions are analyzed with immunocytochemical stain.
(5) the C2C12 multinuclear fleshes of the cell experiment of repeated packets, the adenovirus processing differentiation of 3 kinds of interference BCKDK expression are small
Pipe, real-time qPCR analyses BCKDK, BCKDP, BCKDE1A, MyoD, Myogeinin, MyHC, MuRF1 and MAFbx's
MRNA expressions.
(6) at the same westernblot detection BCKDK, MyoD, Myogeinin, MyHC and BCKDE1A protein expression feelings
Condition.
Immunocytochemical stain result is shown in Fig. 6.(A) the C2C12 multinuclears of the adenovirus processing differentiation of interference BCKDK expression
Myotubule can cause muscle fiber atrophy to attenuate.(B) the adenovirus processing of interference BCKDK expression can cause muscle fibre MyHC expression to subtract
Few, especially the 3rd kind of adenovirus (BCKDK-V3) causes muscle fiber atrophy effect most apparent.
MRNA expression influences result and sees that Fig. 7, the adenovirus processing of 3 kinds of interference BCKDK expression can reduce multinuclear myotubule
BCKDK expression increases BCKDE1A expression, increases E3 ligases MuRF1 and MAFbx and expresses, reduction muscle fibre MyoD,
Myogeinin and MyHC expressions, but there is different characteristic.In general, the adenovirus of the third interference BCKDK
(BCKDK-V3) effect is most notable, for this purpose, subsequent experimental is carried out with the adenovirus (BCKDK-V) of the third interference BCKDK.
Protein expression influences result and sees Fig. 8, after interference BCKDK expression, multinuclear myotubule meeting BCKDK, MyoD,
The protein expression of Myogeinin and MyHC reduces, and BCKDE1A expression increases.
Embodiment 6
The present embodiment is influence of In vivo study BCKDK and the BCKDC change to skeletal muscle atrophy, and experimental method is as follows：
(1) experimental method establishes tumor cachexia animal model as described in Example 2.
(2) tumor cachexia animal pattern is grouped, and is interfered Gastrocnemius Muscle of Cancer local injection using micro syringe within the 7th day
The adenovirus of BCKDK expression.
It puts to death mouse within (3) the 21st days, compares gastrocnemius quality and histopathologic characteristics, Real-timeqPCR analyzes sura
Myogen different differentiation phases marker MyHC, Myogeinin, MyoD of flesh and tibialis anterior；E3 ligases；BCKDK and
BCKDPmRNA expressions change.
Experimental result is shown in Fig. 9, (A) gastrocnemius local injection interferes the adenovirus of BCKDK expression, gastrocnemius part BCKDK
MRNA expression significantly reduce, the mRNA of BCKDP expression is significantly raised, but BCKDK and the BCKDP expression of tibialis anterior are without significantly
Change.(B) Gastrocnemius Muscle of Cancer MyoD protein expressions obviously inhibit after BCKDK expression interference, and MuRF1 protein expressions are significantly raised；
(C) the mRNA expression of Gastrocnemius Muscle of Cancer MyHC, Myogeinin and MyoD also substantially reduces；(D) HE dyeing also shows interference
Cross section diameter of muscle fiber attenuates after BCKDK.
By above-described embodiment it is found that the present invention is from its critical limitation metabolic enzyme of tumor cachexia metabolic regulation angle analysis
Regulatory mechanism, and pharmaceutical intervention target spot is determined：Branched-chain alpha-ketoacid dehydrogenase complex (BCKDC) or branched-chain alpha-keto acid dehydrogenase
Kinases (BCKDK) has potential clinical value in skeletal muscle atrophy treatment.
Specific embodiments of the present invention are described in detail above, but it is only used as example, the present invention is not intended to limit
In particular embodiments described above.To those skilled in the art, it is any to the invention carry out equivalent modifications and replace
In generation, is also all among scope of the invention.Therefore, without departing from the spirit and scope of the invention made by impartial conversion and repair
Change, all should be contained within the scope of the invention.
Claims (10)
1. branched-chain alpha-ketoacid dehydrogenase complex or branched-chain alpha-keto acid dehydrogenase kinases are preventing or are controlling as drug target
Treat the application in skeletal muscle atrophy.
2. branched-chain alpha-ketoacid dehydrogenase complex or branched-chain alpha-keto acid dehydrogenase kinases are preparing prevention or treatment skeletal muscle atrophy
Application in drug.
3. application according to claim 1 or 2, which is characterized in that the skeletal muscle atrophy include Sarcopenia, amyotrophia,
Muscular dystrophy, cachexia；Wherein, the cachexia includes at least chronic heart failure, chronic obstructive pulmonary disease, chronic renal
The cachexia of one or more inductions sick, in rheumatoid arthritis.
4. application according to claim 2, which is characterized in that the application includes that inside and outside screening prevents or treat bone
The drug of amyotrophia comprising following steps：With branched-chain alpha-ketoacid dehydrogenase complex or its gene or branched-chain alpha-ketoacid dehydrogenation
Kinase enzyme or its gene pick out the inhibitor or branch α -one of branched-chain alpha-ketoacid dehydrogenase complex as drug effect object
The activator of acidohydrogenase kinases is as the candidate primary dcreening operation drug for preventing or treating skeletal muscle atrophy.
5. application according to claim 4, which is characterized in that the inside and outside screening prevents or treat skeletal muscle atrophy
The step of drug includes：Using the gene of branched-chain alpha-ketoacid dehydrogenase complex or branched-chain alpha-ketoacid dehydrogenase complex as drug
Effective object, the inhibitor for picking out branched-chain alpha-ketoacid dehydrogenase complex are first as the candidate of prevention or treatment skeletal muscle atrophy
Sieve drug；And using the gene of branched-chain alpha-keto acid dehydrogenase kinases or branched-chain alpha-keto acid dehydrogenase kinases as drug effect pair
As picking out the activator of branched-chain alpha-keto acid dehydrogenase kinases as the candidate primary dcreening operation drug for preventing or treating skeletal muscle atrophy.
6. application according to claim 5, which is characterized in that the inhibitor is to branched-chain alpha-ketoacid dehydrogenase complex
Compound with inhibition, the inhibition include inhibiting branched-chain alpha-ketoacid dehydrogenase complex activity, or inhibit
The genetic transcription or expression of branched-chain alpha-ketoacid dehydrogenase complex.
7. application according to claim 5, which is characterized in that the activator is to have to branched-chain alpha-keto acid dehydrogenase kinases
It includes inhibiting branched-chain alpha-ketoacid dehydrogenase complex activity, or promote branch to have the compound of activation effect, the activation effect
The genetic transcription or expression of chain alpha-ketoacid dehydrogenase kinases.
8. the application described according to claim 6 or 7, which is characterized in that judge whether candidate primary dcreening operation drug can inhibit branch α-
The active method of ketoacid dehydrogenase complex includes enzyme kinetics method.
9. the application described according to claim 6 or 7, which is characterized in that judge whether candidate primary dcreening operation drug can inhibit branch α-
The genetic transcription or expression of ketoacid dehydrogenase complex or the genetic transcription and expression for promoting branched-chain alpha-keto acid dehydrogenase kinases
Method includes the following steps：The thin of normal expression branched-chain alpha-ketoacid dehydrogenase complex or branched-chain alpha-keto acid dehydrogenase kinases is provided
Born of the same parents or mouse cultivate the cell or mouse, detection branch in drug to be measured or in the presence of carrying the carrier of drug to be measured
Whether the transcription of chain alpha-ketoacid dehydrogenase complex or branched-chain alpha-keto acid dehydrogenase kinases and expression change.
10. the application described according to claim 6 or 7, which is characterized in that the inhibition of the branched-chain alpha-ketoacid dehydrogenase complex
The activator of agent and branched-chain alpha-keto acid dehydrogenase kinases at least have the effect that in it is one or more：Inhibit skeletal muscle tissue
E3 ubiquitination ligases MuRF1 and MAFbx/Atrogin-1 expression；Inhibit the ubiquitin protein enzyme body activation of skeletal muscle tissue；
Regulate and control expression and the phosphorylation of the FoxO1 and FoxO3 of skeletal muscle tissue；The mTOR expression of promotion skeletal muscle tissue increases or phosphorylation
Degree increases；Skeletal muscle tissue MyHC, Myf4, Myogeinin, Myf5, MyoD and Pax3 expression is promoted to increase；Increase skeletal muscle
Quality or weight；Increase hand grip endurance.
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