CN107019791A - Non-peptide micromolecular compound as MC4R activators and its specific mutants medicine companion application - Google Patents
Non-peptide micromolecular compound as MC4R activators and its specific mutants medicine companion application Download PDFInfo
- Publication number
- CN107019791A CN107019791A CN201610401872.8A CN201610401872A CN107019791A CN 107019791 A CN107019791 A CN 107019791A CN 201610401872 A CN201610401872 A CN 201610401872A CN 107019791 A CN107019791 A CN 107019791A
- Authority
- CN
- China
- Prior art keywords
- mc4r
- bms
- application
- compound
- mutant
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/05—Dipeptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/06—Dipeptides
- C07K5/06139—Dipeptides with the first amino acid being heterocyclic
- C07K5/06147—Dipeptides with the first amino acid being heterocyclic and His-amino acid; Derivatives thereof
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention belongs to medicine preparation and applied technical field, and in particular to non-peptide micromolecular compound as MC4R activators and its specific mutants medicine companion application.Present invention firstly discovers that a kind of non-peptide micromolecular compound; its numbering is BMS470539; its chemical name is 1 [piperidyl of 1 (3 methyl L histidyl- O methyl D tyrosyls) 4 phenyl 4] 1 butanone, and its molecular weight is 559.697.The compound is a kind of activator of Melanocortin receptor 4 acceptor, can recover some MC4R defects mutant as medicine companion and produce signaling molecule ability.
Description
Technical field
The present invention relates to medicine preparation and applied technical field, and in particular to non-peptide micromolecular compound swashs as MC4R
Dynamic agent and its application of specific mutants medicine companion.Described non-peptide micromolecular Compound nomenclature is BMS-470539, profit
The application for preparing MC4R activators and special MC4R mutant medicine companion is used as with BMS-470539.
Background technology
Melanocortin receptor family includes 5 various physiological functions g protein coupled receptors (melanocortin receptor 1-5)1.It is black
Melanocortin receptor -1 mainly acts on coloring and the anti-inflammatory of melanocyte, the life of the main function of melanocortin receptor -2 steroids
Into melanocortin receptor -3 mainly acts on acquiring rate for food and nutrient distribution, the mainly regulation food intake of melanocortin receptor -4
And energy adjustment, melanocortin receptor -5 acts on outer secretion, particularly cortex glandular secretion2-7.In recent years, adult's obesity rates
Rapid growth turn into influence worldwide in human health major issue.Evidence suggests melanocortin receptor -4
(Melanocortin-4receptor, MC4R) knock-out mice and the MC4R mutation mankind show early stage property obesity and with appetite
Cross Sheng, hyperglycaemia and Hyperinsulinemia8-10.In addition, human genetics research shows MC4R code areas mutation stands most
The single gene mutation frequently occurred leads obesogenous case11-14.Based on external functional study, MC4R mutant is divided into five classes：
The first kind is null mutation acceptor；Equations of The Second Kind is cell membrane localization defect acceptor；3rd class is the affine defect acceptor of part；4th
Class is signal activation defect acceptor；5th class is function burst-normal acceptor15。
Most of nonactive MC4R mutant have a different degrees of cell membrane localization defect, ligand affinity defect,
CAMP signals or pERK1/2 signal defects.Therefore, how to save these functional defects of MC4R mutant turns into treatment obesity
Study hotspot.Such as heat shock protein A subunit can instruct MC4R correctly to fold as molecular chaperones can not be correct so as to save those
The mutant of folding16.Also medicine companion PBA17, small molecule agonist THIQ18, small molecular antagonists ML00025376419All
There is a certain degree of gene therapy to act on.But these molecules or medicine companion are mutated just for some special MC4R defects
Body, therefore, it is necessary to the medicine companion for saving MC4R defect mutant further be excavated, for the new personalized antiadipositas drug of exploitation
Thing lays the foundation.
Non-peptide micromolecular compound BMS-470539 chemical name is 1- [1- (3- methyl-L- histidyl--O- first
Base-D- tyrosyls) -4- phenyl -4- piperidyls] -1- butanone, molecular weight is 559.697, the compound earliest be for MC1R by
The activator of body exploitation16.Identical part can be combined based on MC4R and MC1R, and activate similar signal path.Prompting can be with
For BMS-470539 carry out its as the pharmacological property research of the new activators of MC4R and its as medicine chaperone in itself
Functional study to being saved MC4R mutant20。
The content of the invention
It is an object of the invention to provide a kind of non-peptide micromolecular compound, numbering is BMS-470539 as melanocortin
The application of acceptor -4 (abbreviation MC4R) activator.
It is BMS-470539 as preparing special MC4R mutant medicine it is a further object to provide one kind numbering
The application of thing companion.
The chemical name of above-mentioned BMS-470539 compounds is 1- [1- (3- methyl-L- histidyl-s-O- methyl Ds-junket ammonia
Acyl) -4- phenyl -4- piperidyls] -1- butanone, its chemical structural formula is as shown in figure 1, its molecular weight is 559.697.
In order to screen the active drug for saving MC4R defect mutant, applicant is to non-peptide micromolecular compound BMS-
470539 are studied.Applicant have studied BMS-470539 to the wild receptor actings of MC4R first, find its energy and MC4R
Specifically bind and activate signal path downstream.Then, to develop its redemption effect to MC4R mutant, applicant selects
10 MC4R Equations of The Second Kind mutant (N62S, I69R, P78L, C84R, G98R, Y157S, M161T, W174C, P260Q and
C271Y), 6 MC4R the 3rd class mutant (G55V, Δ 88-92, I102T, L106P, D126Y and A219V) and 4 MC4R
The 4th class mutant (D90N, S136F, A175T and C326R) carried out related drugs save targeting screening test.It was found that
BMS-470539 can the specific signal transduction functionality for saving some of which defect mutant.
Specifically, the present invention is achieved through the following technical solutions：
Applicant is tested by ligand affinity, the activation of cAMP and pERK1/2 signaling molecules detection, finds BMS-
470539 have high-affinity (as shown in Figure 2 and Table 1) with MC4R, and BMS-470539 also induces MC4R in dose-dependent form
Signaling molecule cAMP (as shown in Figure 3 and Table 1) is produced, and induces MC4R to produce high-caliber pERK1/2 (as shown in Figure 4,5).
So as to find that BMS-470539 is MC4R activator.
Applicant has found that BMS-470539 can preferably recover the class mutant of MC4R bis- by the detection to cAMP signal activations
A175T, C326R in middle N62S, C84R, M161T, P260Q, G55V, I102T, L106P in the mutation of three classes, and the mutation of four classes
Produce the ability of cAMP signaling molecules (as shown in Fig. 6,7,8 and table 2,3).
Applicant has found that BMS-470539 can preferably recover three classes mutation G55V by the WB detections to pERK1/2 albumen,
Δ 88-92, I102T, L106P, D126Y, A219V, and four class mutant S136F, A175T, C326R produce pERK1/2 letters
Number molecule ability (as Fig. 9,10 and table 3 shown in).
More detailed technical scheme is such as《Embodiment》It is described.
Brief description of the drawings
Fig. 1：It is non-peptides micromolecular compound BMS-470539 chemical structural formula.
Fig. 2：It is the level that BMS-470539 inductions lower MC1R, MC3R, MC4R and MC5R produce signaling molecule cAMP.
Fig. 3：Be MC1R, MC3R, MC4R and MC5R respectively with BMS-470539 ligand affinity results.
Fig. 4：It is the WB results that MC4R produces signaling molecule pERK1/2 levels under different time BMS-470539 inductions
Figure.
Fig. 5：It is the WB results that MC4R produces signaling molecule pERK1/2 levels under various concentrations BMS-470539 inductions
Figure.
Fig. 6：It is Equations of The Second Kind MC4R mutant 10-5The lower horizontal columns for producing signaling molecule cAMP of M BMS-470539 inductions
Shape figure.
Fig. 7：It is the folding that the 3rd class MC4R mutant produces signaling molecule cAMP under various concentrations BMS-470539 inductions
Line chart.Description of reference numerals：A figures in Fig. 7 are G55V, Δ 88-92, I102T mutant and WT acceptors in various concentrations BMS-
The lower line chart for producing signaling molecule cAMP of 470539 inductions；B figures in Fig. 7 are L106P, D126Y, A219V mutant and WT
Acceptor produces signaling molecule cAMP line chart under various concentrations BMS-470539 inductions.
Fig. 8：It is the folding that the 4th class MC4R mutant produces signaling molecule cAMP under various concentrations BMS-470539 inductions
Line chart.
Fig. 9：It is the 3rd class MC4R mutant 10-5M BMS-470539 inductions are lower to produce signaling molecule pERK1/2 levels
WB result figures.Description of reference numerals：A figures in Fig. 9 are WB detection figures；B figures in Fig. 9 are WB results statistics block diagram.
Figure 10：It is the 4th class MC4R mutant 10-5M BMS-470539 inductions are lower to produce signaling molecule pERK1/2 levels
WB result figures.Description of reference numerals：A figures in Figure 10 are WB figures；B figures in Figure 10 are WB results statistics block diagram.
Embodiment
To understand the present invention, with reference to embodiment, the invention will be further described；Following embodiments be it is illustrative,
It is not limited, it is impossible to limit protection scope of the present invention with following embodiments.
Embodiment 1 prepares embodiment substantially
The present invention chooses commercialization plasmid pcDNA3.1 (+) (being purchased from Clontech companies) as expression vector, constructs open country
The MC4R (hMC4R) of raw type (WT) people expression vector：pcDNA3.1-hMC4R-WT.I.e. in pcDNA3.1 (+) polyclonal position
Point inserts the expressed sequence (NM_005912.2) of an encoding wild type hMC4R gene.
PcDNA3.1-hMC4R-WT of the present invention to build (is purchased from as template using Fast Fixed-point mutagenesis kit
Stratagene companies) construct 20 and be named as N62S, I69R, P78L, C84R, G98R, Y157S, M161T, W174C,
P260Q, C271Y, G55V, Δ 88-92, I102T, L106P, D126Y, A219V, D90N, S136F, A175T, C326R is through function
Identification second and third, four class MC4R mutation body expression vector.
The present invention will not express MC4R acceptor people's kidney embryonic cells (HEK293T) as MC4R and the transient expression of mutant
System, for detecting ligand affinity, cAMP signals and pERK1/2 signals.HEK293T cells (being purchased from ATCC companies) are in 37
DEG C, cell is cultivated in 5%CO2 incubators, cultivating system is the DMEM culture mediums containing 10% calf serum.The day before transfection, will
Cell is with 0.14 × 106Individual/ml density inoculating cell (treats that cell attachment trails), using phosphoric acid into 6 orifice plates after 30h
Calcium method21Sample is received after transfecting MC1R mutant and wild-type positive control plasmid, 48h.
The present invention is related to ligand affinity detection (see embodiment to BMS-470539 as the Function Identification of MC4R activators
2), the detection of cAMP signal activations ability is (see embodiment 3) and produces signaling molecule pERK ability detection (see embodiment 4).
Meanwhile, generation signaling molecule is related to as the redemption Function Identification of MC4R mutant medicine companions to BMS-470539
The ability that cAMP ability detected (see embodiment 5,6) and produced signaling molecule pERK is detected (see embodiment 7).
Detection of the embodiment 2 to BMS-470539 and MC1R, MC3R, MC4R, MC5R affinity
HEK293T cells are received before sample, cell are washed 2 times with serum free medium (being purchased from Gibco companies), by different final concentrations
(10-10~10-5M) radioactive mark ligands of the part BMS-470539 (being purchased from Bachem companies) with 100,000cpm125I-
NDP-MSH (be purchased from peptide radioiodination service centre of University of Mississippi) (50 μ L) 37 DEG C it is common incubate transfection MC1R,
After MC3R, MC4R and MC5R HEK293T cells (being purchased from ATCC companies) 1h, terminating reaction, with the phosphate buffer of precooling
That is PBS (137mM NaCl, 2.7mM KCl, 1.4mM KH2PO4, 4.3mM Na2HPO4, pH 7.4) and wash away the mark not being combined
Remember thing, the 0.5N NaOH for adding 100 μ L are collected after cell, determined and combined in cell with acceptor using gamma scintillator calculating instrument
's125I-NDP-MSH total amount, that is, draw MC1R, MC3R, MC4R and MC5R and part BMS-470539 binding ability.Utilize
GraphPadPrism5 software analysis experimental datas make nonlinear regression S type curves, and calculating obtains binding of receptor and ligand
Median effective concentration value (IC50) and Cmax value (Bmax)。
By Competition binding assay, it is found by the applicant that BMS-470539 to MC1R in addition to very strong affinity is shown, it is right
MC4R acceptors also have at a relatively high affinity, and decline 4 times to MC3R affinity.MC5R is then only 10-5M BMS-
There is faint affinity therewith under 470539 stimulations.As a result Fig. 2 and table 1 are seen.
Table 1 MC1R, MC3R, MC4R, MC5R and BMS-470539 affinity and the generation under BMS-470539 inductions
The feature of cAMP signals
Table 1 illustrates：aRepresent the p compared with MC1R acceptors<0.05；bRepresent the p compared with MC1R acceptors<0.01；cRepresent with
MC1R acceptors compare p<0.001；N/A represents not detect.
The BMS-470539 of embodiment 3 inductions MC1R, MC3R, MC4R and MC5R produce signaling molecule cAMP ability detection
HEK293T cells are received before sample, cell are washed with serum free medium 2 times, and the isobutyl methyl Huang containing 0.5mM is fast
The serum free medium (being purchased from Gibco companies) of purine incubates the HEK293T for having transfected MC1R, MC3R, MC4R and MC5R at 37 DEG C
Cell 15min, then by different final concentrations (10-10~10-5M BMS-470539 stimulation process cells), 37 DEG C incubate after 1h,
Tissue Culture Plate is put on ice, culture medium is abandoned in suction, add the 0.5N perchloric acid containing 180 μ g/mL theophylline and extract intracellular cAMP, profit
With radiommunoassay (RIA) method22The dose dependent for carrying out signaling molecule cAMP generations is determined, and is utilized
GraphPadPrism5 softwares calculate its median effective concentration (EC50) and Cmax value (Rmax)。
The measurement result of the present embodiment is consistent with ligand affinity measurement result, and the experiment of cAMP activity inducements shows, BMS-
470539 decapacitation induction MC1R is produced outside cAMP, moreover it is possible to stimulate MC4R to produce dose dependent cAMP, its EC50For 472.4nM；
RmaxFor 1887.4pmol/106Individual cell.As a result Fig. 3 and table 1 are seen.
The BMS-470539 of embodiment 4 inductions MC4R produces signaling molecule pERK ability detection
Detect that (extracellular regulated protein of phosphorylation swashs pERK using conventional western blot (abbreviation WB) methods
Enzyme) level.After HEK293T cell transfecting wild type MC4R carriers 24h, then with serum free medium (be purchased from Gibco companies)
Make HEK293T cells starvation 24h.Sample day is being received, is being induced with BMS-470539.HEPES buffer solution is first used before receiving sample
【150mM NaCl and 20Mm Hepes (4- hydroxyethyl piperazineethanesulfonic acids), pH 7.4】Wash cell 2 times, then add lysate【HEPES
(Nonidet P40) containing 0.5%NP-40 in buffer solution, 2mM EDTA (ethylenediamine tetra-acetic acid), 1mM Na3VO4(vanadic acid
Sodium) and 1mM NaF (sodium fluoride)】Collect cell.Total protein concentration Bradford methods23Determine.Per hole loading total protein 35mg,
Gone to after being separated with 10%SDS-PAGE glue on pvdf membrane.Pvdf membrane 10% skimmed milk power (containing 0.2%Tween-20) room temperature
Close after 4h, plus two kinds of primary antibodies, 4 DEG C of overnight incubations, (volume ratio is 1 to wherein primary antibody rabbit-anti pERK1/2:And the anti-β of mouse 2000)-
(as internal reference antibody, volume ratio is 1 to tubulin:5000) with containing 5% bovine serum albumin(BSA) be BSA TBST (purchased from Shanghai give birth to
Work bioengineering Co., Ltd) dilute.Second day, plus (volume ratio is 1 to the anti-rabbit of horseradish peroxidase-labeled:1500) and
(volume ratio is 1 to anti-mouse:5000) two anti-igg are incubated 2h at room temperature, and described secondary antibody is diluted with 10% skimmed milk power.Finally use
ECL development processes24Colour developing.Then densitometric scan is carried out to protein band with image processing software ImageJ.ERK1/2 phosphoric acid
Change level is standardized by pERK1/2 and β-tubulin ratio.
As a result show that BMS-470539 can induce MC4R acceptors to produce ERK signals.When BMS-470539 acts on 5min, energy
MC4R is induced to produce the pERK1/2 of 3.7 times of foundation level, but effect 30min to 60min has just fallen back to foundation level, such as
Shown in Fig. 4.From dose dependent result, 10-7M BMS-470539 is with regard to that can induce MC4R to produce 4.0 times of foundation levels
PERK1/2, the BMS-470539 Inducement differences of higher concentration are notable, as shown in Figure 5.
Influences of the BMS-470539 of embodiment 5 to Equations of The Second Kind MC4R mutant cAMP activity
Influences of the BMS-470539 to Equations of The Second Kind MC4R mutant cAMP activity uses RIA methods22Determine, specific steps
For：HEK293T cells receive sample the previous day, with 10-5The BMS-470539 processing of M final concentrations has transfected N62S, I69R, P78L,
The cell of C84R, G98R, Y157S, M161T, W174C, P260Q, C271Y and WT plasmid 24 hours.Second day, cell received sample
Before, cell is washed with serum free medium 2 times, and by the serum free medium of the isobutyl methylxanthine containing 0.5mM at 37 DEG C
Incubate cells 15min, then Jia 10-6M NDP-MSH act on 37 DEG C and incubated after 1h, and Tissue Culture Plate is put on ice, and culture medium is abandoned in suction,
Add the 0.5N perchloric acid containing 180 μ g/mL theophylline and extract intracellular cAMP, utilize RIA methods22Carry out signaling molecule cAMP levels
Determine, and its concentration value is calculated using GraphPadPrism5 softwares.
RIA testing results show that BMS-470539 adds the cAMP levels of N62S, C84R, M161T and P260Q generation,
And I69R, P78L, G98R, Y157S, W174C are had no significant effect, as a result as shown in Figure 6.
Influences of the BMS-470539 of embodiment 6 to third and fourth class MC4R mutant cAMP signals activity
Influences of the BMS-470539 to third and fourth class MC4R mutant cAMP signals activity uses RIA methods22Determine.Tool
Body step is：Cell is received before sample, cell is washed with serum free medium 2 times, and by the nothing of the isobutyl methylxanthine containing 0.5mM
Blood serum medium has transfected G55V, Δ 88-92, I102T, L106P, D126Y, A219V, D90N, S136F in 37 DEG C of incubations,
The HEK293T cell 15min of A175T, C326R and WT plasmid, then by different final concentrations (10-7M~10-5M BMS-)
470539 in 37 DEG C of stimulation process cell 1h.Tissue Culture Plate is put on ice, culture medium is abandoned in suction, added containing 180 μ g/mL theophylline
0.5N perchloric acid extracts intracellular cAMP, utilizes RIA22The dose dependent for carrying out signaling molecule cAMP generations is determined, and is utilized
GraphPadPrism5 softwares calculate its concentration value.
RIA testing results show, BMS-470539 can induce the 3rd class be mutated G55V, I102T and L106P produce dosage according to
Rely property cAMP, and the 4th class of induction mutation A175T and C326R to produce dose dependent cAMP, but other three, four classes are mutated
Have no significant effect, as a result as shown in Fig. 7, Fig. 8 and table 2.
Influences of the BMS-470539 of table 2 to wild type and third and fourth class MC4R mutant cAMP and pERK1/2 signals
The explanation of table 3：“↑”：Show that expression or activity are improved compared with foundation level；“↓”：Show compared with foundation level
Expression or activity reduction；“-”：Show not significantly change compared with foundation level.
Influences of the BMS-470539 of embodiment 7 to third and fourth class MC4R mutant pERK1/2 signals activity
Influences of the BMS-470539 to third and fourth class MC4R mutant pERK1/2 signals activity is detected using WB methods.Tool
Body step is：With HEK293T cell transfecting G55V, Δ 88-92, I102T, L106P, D126Y, A219V, D90N, S136F,
After A175T, C326R and WT plasmid 24h, then with serum free medium make HEK293T cells starvation 24h.Sample day is being received, with 10-5M
BMS-470539 induces 5min.HEPES buffer solution is first used before receiving sample【150mM NaCl and 20Mm Hepes (4- hydroxyethyl piperazines
Ethyl sulfonic acid), pH 7.4】Wash cell 2 times, then add lysate【(the poly- second two of ethylphenyl containing 0.5%NP-40 in HEPES buffer solution
Alcohol), 2mM EDTA (ethylenediamine tetra-acetic acid), 1mM Na3VO4(sodium vanadate) and 1mM NaF (sodium fluoride)】Collect cell.Total protein
Concentration Bradford methods23Determine.Per hole loading total protein 35mg, gone to after being separated with 10%SDS-PAGE glue on pvdf membrane.
Pvdf membrane is closed after 4h with 10% skimmed milk power (containing 0.2%Tween-20) room temperature, plus two kinds of primary antibodies, 4 DEG C of overnight incubations, wherein
Primary antibody is that (volume ratio is 1 to rabbit-anti pERK1/2:2000) (as internal reference antibody, volume ratio is 1 with the anti-β-tubulin of mouse:5000)
Diluted with being BSA TBST (being purchased from Shanghai Sheng Gong bioengineering Co., Ltd) containing 5% bovine serum albumin(BSA)；Second day, plus
(volume ratio is 1 to the anti-rabbit of horseradish peroxidase-labeled:1500) (volume ratio is 1 with anti-mouse:5000) two anti-igg are at room temperature
2h is incubated, described secondary antibody is diluted with 10% skimmed milk power.Finally use ECL development processes24Colour developing.Then image processing software is used
ImageJ carries out densitometric scan to protein band.The ratio that ERK1/2 phosphorylation level passes through pERK1/2 and β-tubulin
It is standardized.
WB testing results show that BMS-470539 can induce all 3rd class MC4R mutant G55V, Δ 88-92,
I102T, L106P, D126Y and A219V produce pERK1/2, and induction the 4th class mutant S136F, A175T and C326R productions
Raw pERK1/2, invention effect is as shown in Fig. 9, Figure 10 and table 2.
Bibliography
1 Chan,L.F.et al.MRAP and MRAP2 are bidirectional regulators of the
melanocortin receptor family.Proceedings of the National Academy of Sciences
106,6146-6151(2009).
2 Roberts,D.W.,Newton,R.A.,Beaumont,K.A.,Helen Leonard,J.&Sturm,
R.A.Quantitative analysis of MC1R gene expression in human skin cell
cultures.Pigment cell research 19,76-89(2006).
3 Beaumont,K.A.et al.Altered cell surface expression of human MC1R
variant receptor alleles associated with red hair and skin cancer risk.Human
molecular genetics 14,2145-2154(2005).
4 Doyle,J.R.,Fortin,J.-P.,Beinborn,M.&Kopin,A.S.Selected melanocortin
1 receptor single-nucleotide polymorphisms differentially alter multiple
signaling pathways.Journal of Pharmacology and Experimental Therapeutics 342,
318-326(2012).
5 Roy,S.,Rached,M.&Gallo-Payet,N.Differential regulation of the human
adrenocorticotropin receptor [melanocortin-2 receptor(MC2R)]by human MC2R
accessory protein isoformsαandβin isogenic human embryonic kidney 293
cells.Molecular Endocrinology 21,1656-1669(2007).
6 Yang,Z.,Huang,Z.-L.&Tao,Y.-X.Functions of DPLIY motif and helix 8
of human melanocortin-3receptor.Journal of molecular endocrinology 55,107-117
(2015).
7 Abdel-Malek,Z.et al.Mitogenic and melanogenic stimulation of normal
human melanocytes by melanotropic peptides.Proceedings of the National
Academy of Sciences of the United States of America 92,1789-1793(1995).
8 Huszar,D.et al.Targeted disruption of the melanocortin-4 receptor
results in obesity in mice.Cell 88,131-141(1997).
9 Farooqi,I.S.et al.Clinical spectrum of obesity and mutations in the
melanocortin 4 receptor gene.New England Journal of Medicine 348,1085-1095
(2003).
10 Yeo,G.S.et al.A frameshift mutation in MC4R associated with
dominantly inherited human obesity.Nature genetics 20,111-112(1998).
11 Loos,R.J.et al.Common variants near MC4R are associated with fat
mass,weight and risk of obesity.Nature genetics 40,768-775(2008).
12 Vaisse,C.et al.Melanocortin-4 receptor mutations are a frequent
and heterogeneous cause of morbid obesity.Journal of Clinical Investigation
106,253-262(2000).
13 Tao,Y.-X.Mutations in Melanocortin‐4 Receptor and Human
Obesity.Progress in molecular biology and translational science 88,173-204
(2009).
14 Albuquerque,D.,Manco,L.&Nóbrega,C.in Obesity 87-106(Springer,
2016).
15 Tao,Y.-X.&Segaloff,D.L.Functional analyses of melanocortin-4
receptor mutations identified from patients with binge eating disorder and
nonobese or obese subjects.The Journal of Clinical Endocrinology&Metabolism
90,5632-5638(2005).
16 Meimaridou,E.et al.The cytosolic chaperone Hsc70 promotes traffic
to the cell surface of intracellular retained melanocortin-4 receptor
mutants.Molecular Endocrinology 25,1650-1660(2011).
17 Granell,S.,Mohammad,S.,Ramanagoudr-Bhojappa,R.&Baldini,G.Obesity-
linked variants of melanocortin-4 receptor are misfolded in the endoplasmic
reticulum and can be rescued to the cell surface by a chemical
chaperone.Molecular Endocrinology 24,1805-1821(2010).
18 Huang,H.&Tao,Y.-X.A small molecule agonist THIQ as a novel
pharmacoperone for intracellularly retained melanocortin-4 receptor
mutants.International journal of biological sciences 10,817-824(2014).
19 Fan,Z.C.&Tao,Y.X.Functional characterization and pharmacological
rescue of melanocortin‐4 receptor mutations identified from obese
patients.Journal of cellular and molecular medicine 13,3268-3282(2009).
20 Herpin,T.F.et al.Discovery of tyrosine-based potent and selective
melanocortin-1 receptor small-molecule agonists with anti-inflammatory
properties.Journal of medicinal chemistry 46,1123-1126(2003).
21 Chen,C.&Okayama,H.High-efficiency transformation of mammalian
cells by plasmid DNA.Molecular and cellular biology 7,2745-2752(1987).
22 Tao,Y.-X.&Segaloff,D.L.Functional characterization of
melanocortin-3 receptor variants identify a loss-of-function mutation
involving an amino acid critical for G protein-coupled receptor
activation.The Journal of Clinical Endocrinology&Metabolism 89,3936-3942
(2004).
23 Ku,H.K.et al.Interpretation of protein quantitation using the
Bradford assay:comparison with two calculation models.Analytical biochemistry
434,178-180,doi:10.1016/j.ab.2012.10.045(2013).
24 Celis,J.E.&Gromov,P.High-resolution two-dimensional gel
electrophoresis and protein identification using western blotting and ECL
detection.Exs 88,55-67(2000)。
Claims (5)
1. structure shown below compound is in the application as melanocortin receptor -4 (MC4R) activator, it is characterised in that
The structural formula of described compound is as follows：
The chemical name of the compound is 1- [1- (3- methyl-L- histidyl-s-O- methyl Ds-tyrosyl) -4- phenyl -4- piperazines
Piperidinyl] -1- butanone, molecular weight is 559.697.
2. application as claimed in claim 1, it is characterised in that：Described application is included as under activation melanocortin receptor -4
Swim signaling molecule cAMP and p-ERK1/2 application.
3. a kind of non-peptide micromolecular compound of structure as claimed in claim 1, it is named as BMS-470539, and its feature exists
The application for preparing the specific mutant medicine companion of melanocortin receptor -4 is used as in the compound.
4. application as claimed in claim 3, it is characterised in that：The compound BMS470539 divides as cAMP signals are prepared
Son produces MC4R mutant N62S, C84R, M161T, P260Q, G55V, I 102T, L106P, A175T, C326R of capability defect
The application of medicine companion.
5. application as claimed in claim 3, it is characterised in that：The compound BMS470539 believes as p-ERK1/2 is prepared
Number molecule produce the MC4R mutant G55V of capability defect, Δ 88-92, I 102T, L106P, D126Y, A219V, S136F,
The application of A175T, C326R medicine companion.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610401872.8A CN107019791B (en) | 2016-06-07 | 2016-06-07 | Application of non-peptide small molecular compound as MC4R agonist and specific mutant drug chaperone thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610401872.8A CN107019791B (en) | 2016-06-07 | 2016-06-07 | Application of non-peptide small molecular compound as MC4R agonist and specific mutant drug chaperone thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107019791A true CN107019791A (en) | 2017-08-08 |
| CN107019791B CN107019791B (en) | 2020-11-06 |
Family
ID=59525055
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610401872.8A Expired - Fee Related CN107019791B (en) | 2016-06-07 | 2016-06-07 | Application of non-peptide small molecular compound as MC4R agonist and specific mutant drug chaperone thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107019791B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113332285A (en) * | 2021-07-26 | 2021-09-03 | 大理大学 | Application of BMS470539 in preparation of medicine for treating Alzheimer's disease |
| CN115137709A (en) * | 2022-06-30 | 2022-10-04 | 重庆医科大学 | Podocyte active targeting reinforced biological homogeneous composite membrane nano-drug delivery system and preparation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101113982A (en) * | 2007-06-21 | 2008-01-30 | 华东理工大学 | Method for screen selecting polypeptides excitomotor of glucagon-like peptide-1 receptor in vitro |
-
2016
- 2016-06-07 CN CN201610401872.8A patent/CN107019791B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101113982A (en) * | 2007-06-21 | 2008-01-30 | 华东理工大学 | Method for screen selecting polypeptides excitomotor of glucagon-like peptide-1 receptor in vitro |
Non-Patent Citations (6)
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113332285A (en) * | 2021-07-26 | 2021-09-03 | 大理大学 | Application of BMS470539 in preparation of medicine for treating Alzheimer's disease |
| CN115137709A (en) * | 2022-06-30 | 2022-10-04 | 重庆医科大学 | Podocyte active targeting reinforced biological homogeneous composite membrane nano-drug delivery system and preparation method and application thereof |
| CN115137709B (en) * | 2022-06-30 | 2023-05-09 | 重庆医科大学 | Podocyte active targeting reinforced biological homogeneous composite membrane nano drug delivery system and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107019791B (en) | 2020-11-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Burke et al. | Dynamic steps in receptor tyrosine kinase mediated activation of class IA phosphoinositide 3-kinases (PI3K) captured by H/D exchange (HDX-MS) | |
| Peng et al. | Opioid receptor expression in human brain and peripheral tissues using absolute quantitative real-time RT-PCR | |
| Ghosh et al. | Structure of human estrogenic 17β-hydroxysteroid dehydrogenase at 2.20 Å resolution | |
| Burstein et al. | Pharmacology of muscarinic receptor subtypes constitutively activated by G proteins | |
| Laniado et al. | Expression and functional analysis of voltage-activated Na+ channels in human prostate cancer cell lines and their contribution to invasion in vitro. | |
| Gómez-Serrano et al. | Differential proteomic and oxidative profiles unveil dysfunctional protein import to adipocyte mitochondria in obesity-associated aging and diabetes | |
| Imig et al. | Afferent arteriolar dilation to 11, 12-EET analogs involves PP2A activity and Ca2+-activated K+ Channels | |
| Hofmann et al. | Phoenixin is negatively associated with anxiety in obese men | |
| Tan et al. | Quantitative autoradiographic studies of relaxin binding in rat atria, uterus and cerebral cortex: characterization and effects of oestrogen treatment | |
| CN106999541A (en) | Peptidomimetic macrocyclic compound and application thereof | |
| Liang et al. | Pharmacological effect of human melanocortin-2 receptor accessory protein 2 variants on hypothalamic melanocortin receptors | |
| CN101903010B (en) | Topical compositions comprising non-proteogenic amino acids and methods of treating skin | |
| JP2018531938A (en) | Method for treating melanocortin-4 receptor pathway related disorders | |
| Liu et al. | Oral isoflavone supplementation on endometrial thickness: a meta-analysis of randomized placebo-controlled trials | |
| Li et al. | Investigation of triamterene as an inhibitor of the TGR5 receptor: Identification in cells and animals | |
| CN107019791A (en) | Non-peptide micromolecular compound as MC4R activators and its specific mutants medicine companion application | |
| Deigin et al. | Novel platform for the preparation of synthetic orally active peptidomimetics with hemoregulating activity. II. Hemosuppressor activity of 2, 5-diketopiperazine-based cyclopeptides | |
| Wang et al. | A novel peptide exerts potent immunosuppression by blocking the two-site interaction of NFAT with calcineurin | |
| Aissaoui-Zid et al. | AaTs-1: A tetrapeptide from Androctonus australis scorpion venom, inhibiting U87 glioblastoma cells proliferation by p53 and FPRL-1 up-regulations | |
| Anders et al. | Direct identification of the agonist binding site in the human brain cholecystokininB receptor | |
| Kuroda et al. | Determination of topological structure of ARL6ip1 in cells: identification of the essential binding region of ARL6ip1 for conophylline | |
| Hossain et al. | Chemical synthesis and orexigenic activity of rat/mouse relaxin-3 | |
| Selkirk et al. | Identification of differential melanocortin 4 receptor agonist profiles at natively expressed receptors in rat cortical astrocytes and recombinantly expressed receptors in human embryonic kidney cells | |
| Yang et al. | Third transmembrane domain of the adrenocorticotropic receptor is critical for ligand selectivity and potency | |
| Prévost et al. | Mutational and cysteine scanning analysis of the glucagon receptor N-terminal domain |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20201106 |