CN107011427A - Adjust polypeptide of energetic supersession and application thereof - Google Patents
Adjust polypeptide of energetic supersession and application thereof Download PDFInfo
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- CN107011427A CN107011427A CN201710156032.4A CN201710156032A CN107011427A CN 107011427 A CN107011427 A CN 107011427A CN 201710156032 A CN201710156032 A CN 201710156032A CN 107011427 A CN107011427 A CN 107011427A
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- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
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- C07K14/4728—Calcium binding proteins, e.g. calmodulin
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Abstract
The present invention relates to the polypeptide of regulation energetic supersession and its prepare be used to treating with energetic supersession (be particularly fat metabolism) exception about disease medicine in purposes.The polypeptide of the present invention can reduce fat absorption, and the Fat Accumulation in reduction liver simultaneously adjusts fat metabolism, and have the advantages that with general polypeptide products compared with can with oral administration, therefore can also as the health products for being used to adjust fat metabolism active ingredient.
Description
Technical field
The present invention relates to biologic medical field, it is used to treat more particularly to the polypeptide for adjusting energetic supersession and its preparing
With abnormal energy metabolism, particularly fat metabolism extremely about disease medicine in purposes.
Background technology
Obesity is health problem main and growing in world wide.Obesity is also to induce many common diseases (as moved
Pulse atherosclerosis, hypertension, type ii diabetes, dyslipidemia, coronary heart disease, osteoarthritis and Several Kinds of Malignancy) occur danger
Dangerous factor.It can also be by reducing locomitivity and the problem of quality of life causes more serious.Fat generation and drawn by obesity
Increase in all developed country of disease risen.
Fatty liver refers to the excessive caused lesion of fat accumulation in liver cell.The pathogenic factor of fatty liver has many kinds, than
Such as excessive drinking, diet are unreasonable, sitting.It increasingly increases in the incidence of disease of China, as the big threat for threatening health of people.
However, currently used for treatment is fat and its caused disease, including NASH medicine and method all
Still need to develop more effective and low side effect with certain deficiency, therefore for the treatment of fat and NASH
Medicine and method.
BGP is a kind of vitamin k-dependent calbindin for being synthesized and being secreted by Gegenbaur's cell, a kind of non-collagen
Vitamin k-dependent glutaminic acid residue in acidoglycoprotein, its molecule is BGP and Ca2+With reference to critical function base
Group[1,2]。
The content of the invention
The present inventor is it has been surprisingly discovered that a kind of can adjust the polypeptide of energetic supersession from BGP；It has
The function such as blood fat and reduction fat cell volume in fat absorption, reduction liver fat, peripheral blood is reduced, can be effectively clear
Except the fat accumulated in liver cell, reduction blood lipid level and diminution body fat tissue, for treatment fat hepatitis, adjust
Section hypertension and other angiocardiopathies work.
Some aspects of the present invention are related to the polypeptide of regulation energetic supersession, and the polypeptide is by formula M1-Za-M2Represent, wherein：
M1、M2Be each independently with no more than 5, the polypeptide section of 4,3,2 or 1 amino acid residues or be not present；
ZaFor Tyr-Leu-X1-X2-X3-X4-Gly-Ala-X5-X6-Pro-X7-Pro-Asp-X8- Leu-Glu-Pro, its
In：
X1For Tyr, Asn, Asp or it is not present,
X2For Gln, Asn, His, Pro, Ser or it is not present,
X3For Trp, Gly or it is not present,
X4For Leu or it is not present,
X5For Pro or Ser,
X6For Ala or Val,
X7For Tyr or Ser,
X8For Thr or Pro,
And the ZaOptionally there is middle amino acid substitution, insertion or missing and the amino acid substitution, insertion
4, preferably more than 3, more preferably no more than 2, most preferably not more than 1 are no more than with the sum of missing.
Further, in some embodiments of the invention, the Z in the polypeptideaSelected from one below：
SEQ ID NO.1：YLGASVPSPDPLEP,
SEQ ID NO.2：YLYQWLGAPVPYPDPLEP
SEQ ID NO.4：YLNNGLGAPAPYPDPLEP,
SEQ ID NO.5：YLYQWLGAPVPYPDTLEP,
SEQ ID NO.6：YLYQWLGAPVPYPDPLEP,
SEQ ID NO.7：YLDHWLGAPAPYPDPLEP,
SEQ ID NO.8：YLDPGLGAPAPYPDPLEP,
SEQ ID NO.9：YLDHGLGAPAPYPDPLEP,
SEQ ID NO.10：YLDQGLGAPAPAPDPLEP,
SEQ ID NO.11：YLDSGLGAPVPYPDPLEP.
Further, in other embodiments of the invention, the M in the polypeptide1、M2It is not present；Or another
In a little embodiments, Z in the polypeptideaFor YLYQWLGAPVPYPDPLEP, M1It is not present and M2For the amino of Arg, the i.e. polypeptide
Acid sequence is as shown in SEQ ID NO.3；In other embodiment, Z in the polypeptideaFor YLGASVPSPDPLEP, M1Do not deposit
And M2For Thr, the i.e. polypeptide amino acid sequence as shown in SEQ ID NO.18.
The present invention other aspect be directed to adjust energetic supersession polypeptide, its include selected from following any sequence extremely
Lack 6 continuous amino acids and its total amino acid residues is no more than 18, such as no more than 17,16,15,14：
SEQ ID NO.1：YLGASVPSPDPLEP,
SEQ ID NO.2：YLYQWLGAPVPYPDPLEP
SEQ ID NO.3：YLYQWLGAPVPYPDPLEPR,
SEQ ID NO.4：YLNNGLGAPAPYPDPLEP,
SEQ ID NO.5：YLYQWLGAPVPYPDTLEP,
SEQ ID NO.6：YLYQWLGAPVPYPDPLEP,
SEQ ID NO.7：YLDHWLGAPAPYPDPLEP,
SEQ ID NO.8：YLDPGLGAPAPYPDPLEP,
SEQ ID NO.9：YLDHGLGAPAPYPDPLEP,
SEQ ID NO.10：YLDQGLGAPAPAPDPLEP,
SEQ ID NO.11：YLDSGLGAPVPYPDPLEP.
Further, it is comprising following any：
SEQ ID NO.12：PVPYPDPLEP,
SEQ ID NO.13：PYPDPLEP,
SEQ ID NO.14：PDPLEP,
SEQ ID NO.15：SVPSPDPLEP,
SEQ ID NO.16：PSPDPLEP.
Further, it is
SEQ ID NO.12：PVPYPDPLEP,
SEQ ID NO.13：PYPDPLEP,
SEQ ID NO.14：PDPLEP,
SEQ ID NO.15：SVPSPDPLEP,
SEQ ID NO.16：PSPDPLEP.
Some aspects of the present invention are related to the officinal salt of the polypeptide of the present invention.
Some aspects of the present invention are related to such polypeptide of the present invention or its officinal salt, and it, which has, reduces fat absorption,
Blood lipid level is reduced, mitigates NASH and reduces the effect such as body fat tissue.
Some aspects of the present invention are related to polynucleotides, and it encodes hereinbefore polypeptide.
Some aspects of the present invention are related to carrier, and it includes foregoing polynucleotides.
Some aspects of the present invention are related to host cell, and it, which is transfected, has foregoing carrier and can be can marking protein
Under conditions of produce the present invention polypeptide.
The present invention some aspect be related to pharmaceutical composition, its comprising therapeutically effective amount foregoing polypeptide of the present invention or its can
Pharmaceutical salts.
Polypeptide described in the present invention or its officinal salt or pharmaceutical composition have regulation energetic supersession, particularly fat
The effect of fat metabolism.
The polypeptide or its officinal salt or pharmaceutical composition that some aspects of the present invention are related to the present invention are being prepared for controlling
Treat with energetic supersession (more preferably fat metabolism) exception about disease medicine in purposes, the disease be can benefit from
Fat absorption reduce, blood fat reduction, fat consumption increase or Fat Accumulation reduction disease, for example obesity, type ii diabetes,
NASH, insulin resistance, hypertriglyceridemia, hyperglycaemia, high cholesterol, atherosclerosis, coronary heart disease.
Some aspects of the present invention be related to treat with fat metabolism extremely about disease method, it is included to thus needing
The object wanted applies the polypeptide of the invention or its officinal salt or pharmaceutical composition of therapeutically effective amount, and the disease is to be benefited
In the disease of fat absorption reduction, blood fat reduction, fat consumption increase or Fat Accumulation reduction, such as obesity, II type glycosurias
Disease, NASH, insulin resistance, hypertriglyceridemia, hyperglycaemia, high cholesterol, atherosclerosis, coronary disease
Disease.
In some embodiments, polypeptide of the invention or its officinal salt or pharmaceutical composition can become known for any
Treat with fat metabolism extremely about disease medicine and Combination of Methods thing together with apply.
In some embodiments, polypeptide of the invention or its officinal salt or pharmaceutical composition are used to treat and fatty generation
Thank to abnormal relevant disease, the disease is reduced for that can benefit from fat absorption, blood fat is reduced, fat consumption increases or fat product
The few disease of regressive, such as obesity, type ii diabetes, NASH, insulin resistance, hypertriglyceridemia,
High cholesterol, atherosclerosis, coronary heart disease.
The polypeptide or its officinal salt that some aspects of the present invention are related to the present invention are preparing the health care for losing weight
Purposes in product.
Some aspects of the present invention are related to health products for losing weight, its polypeptide comprising the present invention or its is pharmaceutically acceptable
Salt.
In some embodiments, polypeptide of the invention or its officinal salt or pharmaceutical composition can pass through various routines
Mode apply, preferred oral administration.
The polypeptide of the present invention can have a direct impact to the absorption of fat, and oral administration reduces conventional polypeptide administration
Inconvenience, the residue of polypeptide seldom, greatly reduces cost, the potential advantages with the highly significant as medicine.
Brief description of the drawings
Fig. 1 shows that feeding C57BL/6 mouse using high lipid food obtains fat and NASH mould after 12 weeks
Type (Diet-Induced-Obesity＆non-alcoholic fatty liver disease, DIO-NAFLD) with it is normal right
According to weight ratio compared with；Normal diet (ND) and high fat diet (HFD).
Fig. 2 shows that the continuous 6 weeks DIO-NAFLD mouse to high fat diet inject the ISAP of various concentrations1And OCN
After (mouse BGP), compared with control group (only high fat diet mouse), ISAP is injected1Influence to mouse.(A) ND is compareed
Group, HFD control groups and daily intraperitoneal injection OCN or ISAP1The change of group mouse epididymis fat pad；(B) epididymal adipose tissues in each group
Pad average quality；(C) haematoxylin ＆ eosin (H＆E) dyeing of each group epididymal adipose tissues pad histotomy；(D) according to H＆ under double-blind conditions
E stained slices calculate the average fat cell surface product obtained.Statistical analysis：It is compared with HFD control groups.*:P<
0.05,**:P<0.01,***:P<0.001。
Fig. 3 shows that the continuous 6 weeks DIO-NAFLD mouse to high fat diet inject the ISAP of various concentrations1
After OCN, compared with control group (only high fat diet mouse), ISAP is injected intraperitoneally1Influence to mouse liver.(A) each group
The photo of the representativeness image of mouse liver outward appearance；(B) each group mouse liver histotomy H＆E is dyed；(C) each group mouse liver
The surface area of the accumulation NASH cell obtained after oil red O stain under double-blind conditions.Statistical analysis：For with HFD
Control group is compared.*:P<0.05,**:P<0.01,***:P<0.001.Each group has 6 mouse to be used to analyze respectively.
Fig. 4 shows that the continuous 6 weeks DIO-NAFLD mouse to high fat diet inject the ISAP of various concentrations1And OCN
Afterwards, compared with control group (only high fat diet mouse), ISAP is injected1Influence to mouse liver function and blood fat.(A) paddy third
Transaminase (ALT)；(B) alkaline phosphatase (ALP)；(C) glutamic-oxalacetic transaminease (AST)；(D) serum total cholesterol (TC)；(E) it is low close
Spend lipoprotein (LDL)；(F) HDL (HDL).Statistical analysis：*:It is compared with HFD control groups.*:P<
0.05,**:P<0.01,***:P<0.001.#：It is compared with ND control groups.#:P<0.05,##:P<0.01,###P<
0.001.Each group has 6 mouse to be used to analyze respectively.
Fig. 5 shows that gavage applies ISAP1Influence with OCN to small intestine fat absorption.The wild type C57BL/6 of ND feedings
After mouse stomach sterilizing olive oil (30 minutes), ISAP is injected intraperitoneally1With after OCN 30 minutes euthanasia mouse and dissect acquisition it is small
Intestines, carry out frozen section to nearly duodenum section jejunum and use oil red O stain, haematoxylin is redyed (A)；To oil red under double-blind conditions
O positive regions carry out ratio (B) that is quantitative and calculating the area and the intestinal villi gross area by ImageJ softwares.Count credit
Analysis：Compared with physiological saline+olive oil group.***:p<0.001.N=3.
Fig. 6 shows ISAP1And ISAP2Respectively and people GPRC6A combination and receptor internalisation.(A) people GPRC6A is in Hela
It is overexpressed in cell；(B)ISAP1, OCN and OCN-22；And ISAP2, hOCN and hOCN-22 and huGPRC6A be overexpressed
The Binding experiment of the cell membrane of Hela cells；(C) OCN (Cy5-OCN) marked with Cy-5, the OCN-22 marked with Cy-5
(Cy5-Ocn22) ISAP, marked with Cy-52(Cy5-ISAP2) GPRC6A be overexpressed Hela cells in in GPRC6A
The influence of change.
Fig. 7 is shown is applying OCN, ISAP through gavage1、ISAP2、ISAP3Influence in seven weeks after to mouse：(A) mouse
Changes of weight；(B) in stool in mice triglycerides comparision contents.
Fig. 8 shows the comparison of the sequence of the ISAP from different plant species.
Fig. 9 shows that gavage applies ISAP4、ISAP5、ISAP6Influence to small intestine fat absorption.The wild type of ND feedings
C57/b mouse 15, are divided into 5 groups, every group 3, three experimental groups, and gavage applies ISAP respectively4、ISAP5、ISAP6；Two groups pairs
According to, daily gavage applies physiological saline, continues after one week, three groups of experimental groups and a control group gavage administration of sterile olive oil,
One control group gavage applies physiological saline, is euthanized after 20 minutes and mouse and dissects acquisition small intestine, empty to nearly duodenum section
Intestines carry out frozen section and use oil red O stain, and haematoxylin is redyed；It is soft by ImageJ to Oil red O positive area under double-blind conditions
Part carries out ratio that is quantitative and calculating the area and the intestinal villi gross area.Statistical analysis：With physiological saline+olive oil group phase
Compare.***:p<0.001.N=3.
Embodiment
Definition
Term " polypeptide of regulation energetic supersession " used herein is also referred to as " insulin secretion related polypeptide (Insulin
Secretion Association Peptide, ISAP) " refer to from BGP or can be adjusted from derived from BGP
The polypeptide and its variant of amount of energy saving metabolism.
Term as used herein " conserved amino acid replacement " refers to use another amino acid residue with similar quality
The replacement carried out to original acid sequence.For example, lysine residue, arginine residues and histidine residues are with basic side chain
Aspect is similar.In addition, asparagicacid residue and glutaminic acid residue are similar in terms of with acid side-chain.In addition, day
Winter amide residues, glutamine residue, serine residue, threonine residues, tyrosine residue and cysteine residues with
Be in terms of uncharged polar side chain it is similar, and glycine residue, alanine residue, valine residue, leucine residue,
Isoleucine residues, proline residue, trp residue, phenylalanine residue and methionine residues are with non-polar sidechain
Aspect is similar.In addition, tyrosine residue, phenylalanine residue, trp residue and histidine residues are with aromatics side
It is similar in terms of chain.Therefore, it will be apparent for a person skilled in the art that in the amino acid group with above-mentioned similar quality
The amino acid substitution of middle progress will not cause any change of property.
Amino acid used herein is write a Chinese character in simplified form and Chinese control is as follows
Term as used herein " with fat metabolism relevant disease extremely " refers to have heredity or environment or the two is common
The caused disease being characterized with fat metabolic disturbance or its complication, such as, but not limited to obesity, type ii diabetes, non-wine
Essence fatty liver, insulin resistance, hypertriglyceridemia, high cholesterol, atherosclerosis, coronary heart disease.
Term as used herein " NASH " refer to caused by the factor in addition to alcohol with liver cell
Fatty over-deposit is the clinical pathology syndrome of principal character.
Term as used herein " insulin resistance " refers to thin because of the function reduction of the insulin for reducing blood glucose
Born of the same parents can not active combustion glucose state.When insulin resistance is high, human body just generates excessive insulin, and causes high blood
Pressure, conisting of dyslipidemia, heart disease and diabetes etc..Especially in the case of type ii diabetes, due to muscle and adipose tissue
The increase of None- identified insulin, so insulin does not work.
The term used herein being not exactly defined has the implication that those skilled in the art are generally understood.
Specific embodiment
DMEM used in the present invention is purchased from Sigma, wherein supplementing 10%FBS (tire ox bloods in per 500ml culture mediums
Clearly), 1% nonessential amino acid, 1g glucose, 0.75g sodium acid carbonates, 0.1g bovine serum albumin(BSA)s and 1.5ml HEPES (4-
Hydroxyethyl piperazineethanesulfonic acid).
3T3L1 is bought from ATCC.
ISAP1Sequence is Tyr-Leu-Gly-Ala-Ser-Val-Pro-Ser-Pro-Asp-Pro-Leu-Glu-Pro-
Thr(SEQ ID NO.18)。ISAP2Sequence be Tyr-Leu-Tyr-Gln-Trp-Leu-Gly-Ala-Ser-Val-Pro-
Ser-Pro-Asp-Pro-Leu-Glu-Pro(SEQ ID NO.2)。ISAP3Sequence be Tyr-Leu-Tyr-Gln-Trp-
Leu-Gly-Ala–Ser-Val-Pro-Ser-Pro-Asp-Pro-Leu-Glu-Pro-Arg(SEQ ID NO.3)。ISAP4Sequence
It is classified as Ser-Val-Pro-Ser-Pro-Asp-Pro-Leu-Glu-Pro (SEQ ID NO.15).ISAP5Sequence is Pro-Ser-
Pro-Asp-Pro-Leu-Glu-Pro(SEQ ID NO.16)。ISAP6Sequence is Pro-Asp-Pro-Leu-Glu-Pro (SEQ
ID NO.14)。
All zooperies are ratified by the animal welfare committee of advanced technology research institute of the Chinese Academy of Sciences, according to human relations
The requirement of the reason committee is carried out.
Embodiment 1
ISAP1Functional study
1st, high fat diet sets up fat and nonalcoholic fatty liver model
C57BL/6SPF grades of mouse of healthy 6 week old male are bought from Guangdong Province's Experimental Animal Center, weight 18-22g.Point
Two groups, raise in the advanced research institute SPF grades of Animal Houses in Shenzhen.Control group：Give mouse chow diet (ND) (chow diet：Fat
Fat, 5%；Carbohydrate, 53%；Albumen, 23%；Total amount of heat 25J/g), freely ingest and drink water, feed 12 weeks.Experimental group：
Mouse high lipid food (HFD) (high lipid food D12451, Research Diets, Inc.) is given, freely ingests and drinks water, is fed
12 weeks.Detect its physical signs.The mouse weight that high lipid food is fed is more than 40g, it is believed that fat and NASH mould
Type (DIO-NAFLD) is successfully constructed.Fig. 1 is shown in the contrast of mouse weight and control group in structure, and arrow meaning is to start feeding
The time point of high lipid food.
2nd, ISAP is injected intraperitoneally1Influence to DIO-NAFLD mouse
2.1 intraperitoneal injection ISAP1Influence to epididymal adipose tissues pad
When blood glucose is higher than 10mMol, behavior is entered to DIO-NAFLD mouse more than 40g for the mouse weight that high lipid food is fed
The ISAP of 6 weeks phases1Intraperitoneal injection, every group has 6 mouse respectively, by ISAP1It is dissolved in 0.01%BSA normal saline solutions, often
Day, 2pmol/g dosage was injected intraperitoneally into DIO-NAFLD mouse once with 20pmol/g, OCN (Mouse Bone calcium fibroin) with
ISAP1Similar, concentration is 6pmol/g.After 6 weeks, using 95%CO2Mouse is implemented to be euthanized, its epididymal adipose tissues pad group is gathered
Knit, weigh；Then paraffin section is done, fat cell area is observed by microscopy.
Experimental result is shown in Fig. 2, the entirety of (A) ND control groups, HFD control groups and daily intraperitoneal injection polypeptide group mouse adipose
Outward appearance, compared with HFD control groups, using ISAP1Significantly reduce adipose tissue；(B) epididymal adipose tissues of each group mouse are leveled up
Quality, compared with HFD control groups, using ISAP1Significantly reduce weight of epididymal fat pad；(C) each group epididymal adipose tissues pad H＆E contaminates
Color, is disclosed compared with HFD control groups, using ISAP1Fat cell is obviously reduced；(D) cut under double-blind conditions according to H＆E dyeing
Piece calculates the average single fat cell surfaces product obtained, illustrates ISAP1Fat cell is obviously reduced.Statistical analysis：With
HFD control groups are compared.*:P<0.05,**:P<0.01,***:P<0.001.
2.2 intraperitoneal injection ISAP1Influence to liver
In step 2.1, liver is gathered while adipose tissue is gathered, outward appearance is carried out and takes pictures, partial liver group is taken afterwards
Carry out frozen section is knitted, using microscope observation is carried out after oil red O stain, quantitative analysis, experiment are then carried out to equal area
As a result referring to Fig. 3.(A) the representativeness image of each group mouse liver outward appearance, color is lighter, and explanation fat content is higher；HFD+ supporting agents
Cause significant liver fat to accumulate, ISAP is injected intraperitoneally1Significantly reduce Fat Accumulation (B) each group mouse liver H＆ of liver
E is dyed, and light color represents the intracellular fat for dyeing；(C) storage obtained after each group mouse liver oil red O stain under double-blind conditions
The surface area of product NASH cell, it is seen that in liver, ISAP1Compared with HFD is compareed, the ratio shared by fat shows
Reduction is write, shows ISAP1Reduce the fat content in liver cell.
2.3 intraperitoneal injection ISAP1Influence to mouse liver function and blood fat.
In step 2.1, put to death mouse before, tail vein blood, using Roche blood glucose meter (model cobas8000) according to
Specification detection glutamic-pyruvic transaminase (ALT), alkaline phosphatase (ALP), glutamic-oxalacetic transaminease (AST), the serum cholesterol of manufacturer,
And low-density lipoprotein (LDL) and HDL (HDL).Test and result is referring to Fig. 4, or even in 2pmol/g agent
Under amount, compared with HFD is compareed, ISAP1Effectively reduce ALT, ALP, AST, cholesterol and LDL.
Result more than can be seen that ISAP1Intraperitoneal injection the metabolism of the body fat of mouse can be produced it is notable
Influence, reduces the Fat Accumulation in liver cell and fat cell, improves fat metabolism, effectively alleviates NASH
Progress.
3、ISAP1And people GPRC6A combination
3.1 are overexpressed hGPRC6A in Hela cells
1) plating cells：By Hela cell suspensions with 1.6 × 105/ mL density bed board is in 6 orifice plates, per hole 2mL
DMEM culture mediums, in 37 DEG C, 5% CO2Middle culture 24 hours.
2) using Lipofectamine2000 (Invitrogen), hGPRC6 is overexpressed according to the specification of manufacturer and carried
Body (pReceiver-M61) is transfected into Hela cells, changes normal incubation medium within 4 hours after transfection.
3) 48 hours after transfecting, culture medium is discarded, 300 μ L Trizol is added with sterile PBS plate floor cells twice
Solution, extracts cell RNA according to RNAiso Plus (TaKaRa) specification, after being handled through DNA enzymatic, uses SuperScriptT
(Invitrogen companies, Canada) kit is through DNAEngine reverse transcriptions into cDNA.Using cDNA as template, SYBRGreen is used
Method carries out Real Time Monitoring (Light Cycler Roche companies, Germany) to the fluorescence PCR products at each time point, to detect
HGPRC6A expression quantity.If it is desired, obtaining stable expression cell strain using puromycin screening, detected afterwards using qPCR
Its gene expression, as shown in Figure 6A, people GPRC6A stablizes great expression to experimental result in Hela.
The Binding experiment of the cell membrane of Hela cell of the 3.2OCN not homopolypeptide with being overexpressed hGPRC6A
Experimental result is shown in Fig. 6 B, compared with OCN, ISAP1With almost consistent film combination ability, though and OCN-22 can and
GPRC6A is combined but compatibility has differed 10 times, it was demonstrated that ISAP1It is OCN Core domain, with acceptor hGPRC6A phase interactions
With.
4、ISAP1Influence of the acute gavage in normal mouse to small intestine fat absorption
Using the male wild type C57BL/6 mouse of 6 week old, point four groups, every group 3, the interior perfusion sterilizing olive oil solution of enteron aisle
200 μ L (Sigma), after 30 minutes, are injected intraperitoneally OCN and ISAP1(concentration is 6pmol/g), and mouse was put to death in 30 minutes, receive
Collect small intestine sample.Sample be duodenum to caecum section, take isometric 3 sections.Physiology salt close to duodenum section precooling is water-soluble
After liquid is rinsed, frozen section is carried out, fat absorption is observed using oil red O stain.
Experimental result is referring to Fig. 5；(A) frozen section is carried out to nearly duodenum section jejunum and uses oil red O stain, haematoxylin
Redye；(B) carry out quantitative by ImageJ softwares to Oil red O positive area under double-blind conditions and calculate the area and intestinal villi
The ratio of the gross area.As can be seen that relative to saline control, ISAP1It is very effective to reduce olive oil absorption, or even ratio
OCN effect is also substantially a lot.
5th, gavage applies ISAP1Influence to HFD mouse
Using the male wild type C57BL/6 mouse of 6 week old, divide 6 groups, every group 6, the 1st group is given chow diet (ND), 2-
6 groups are given high lipid food (HFD), the 2nd group as control, 3-6 groups are experimental group, daily respectively with 2pmol/g body weight gavages
Using OCN, ISAP1、ISAP2、ISAP3, continue seven weeks, body weight monitored weekly is once.Experimental result is shown in Fig. 7 A, can be with from figure
, it is evident that relative HFD control groups, ISAP1Group occurs in that obvious body weight reduction.*:P<0.05,**:P<0.01,***:P<
0.001。
Collect the stool in mice of the 7th week, toasted 3 days at 60 DEG C, it is ensured that drying, then take 1 milligram, with 1ml chloroforms and
The mixed solution immersion of methanol 2 to 1, then crushes excrement with tissue Syrup-homogenizing instrument, supernatant is taken after centrifugation, examined with Roche blood biochemistry instrument
Its triglycerides is surveyed, experimental result is referring to Fig. 7 B.**:P<0.01,***:P<0.001.Compared with HFD control groups, ISAP1Group
The content of triglycerides is significantly raised in excrement, shows that gavage applies ISAP1Significantly reduce the absorption of triglycerides in enteron aisle.
Embodiment 2
ISAP2Functional study
1、ISAP2And people GPRC6A combination
1.1 are overexpressed hGPRC6A in Hela cells
1) plating cells：By Hela cell suspensions with 1.6 × 105/ mL density bed board is in 6 orifice plates, per hole 2mL
DMEM culture mediums, in 37 DEG C, 5% CO2Middle culture 24 hours.
2) using Lipofectamine2000 (Invitrogen), hGPRC6 is overexpressed according to the specification of manufacturer and carried
Body (pReceiver-M61) is transfected into Hela cells, changes normal incubation medium within 4 hours after transfection.
3) 48 hours after transfecting, culture medium is discarded, 300 μ L Trizol is added with sterile PBS plate floor cells twice
Solution, extracts cell RNA according to RNAiso Plus (TaKaRa) specification, after being handled through DNA enzymatic, uses SuperScriptT
(Invitrogen companies, Canada) kit is through DNAEngine reverse transcriptions into cDNA.Using cDNA as template, SYBRGreen is used
Method carries out Real Time Monitoring (Light Cycler Roche companies, Germany) to the fluorescence PCR products at each time point, to detect
HGPRC6A expression quantity.If it is desired, obtaining stable expression cell strain using puromycin screening, detected afterwards using qPCR
Its gene expression, as shown in Figure 6A, people GPRC6A stablizes great expression to experimental result in Hela.
2、ISAP2With hOCN and the cell membrane for the Hela cells for being overexpressed hGPRC6A Binding experiment
Experimental method with it is essentially identical described in embodiment, experimental result is shown in Fig. 6 B, compared with hOCN, ISAP2With several
Consistent film combination ability, and hOCN-22 does not possess this ability, the in conjunction with the embodiments result of 1 middle term 3.2, it was demonstrated that ISAP1
And ISAP2It is OCN and hOCN Core domain respectively, is interacted with acceptor hGPRC6A, it is thus regarded that ISAP1With
ISAP2With identical function, follow-up signal transduction and biological event are caused by hGPRC6A.
2.3 OCN, hOCN22 marked with Cy5, ISAP2Promote GPRC6A in the Hela cells that GPRC6A is overexpressed
Internalization.
HGPRC6AHela cell suspensions will be expressed with 1.6 × 105/ mL density bed board is being pre-placed gelatin coating lid
In 24 orifice plates of slide, per hole 0.5mL DMEM, in 37 DEG C, 5% CO2Middle culture 24 hours.Cell before treatment, with without blood
After clear culture medium starvation 4h, 100nM Cy5-OCN, Cy5-hOCN22, Cy5-ISAP are added per hole2, 37 DEG C are incubated 30 minutes,
Cell 30 minutes are fixed using poly methanol, and add Triton X-100 (sigma) and are incubated 10 minutes, DAPI (Sigma) 10
Second, according to the specification of manufacturer, to cell and dyeing, is observed and taken pictures using confocal microscope, experimental result is referring to figure
6C.It can be seen that, Cy5-OCN and Cy5-ISAP2Portion is distributed in the cell, and Cy5-hOCN22 is then distributed in extracellular, illustrates again
Cy5-OCN and Cy5-ISAP2It can enter intracellular with acceptor knot merga pass internalization effect, so as to play regulation energy
The effect of metabolism.
2.4 gavages apply ISAP2Influence to HFD mouse
Using the male wild type C57BL/6 mouse of 6 week old, divide 6 groups, every group 6, the 1st group is given chow diet (ND), 2-
6 groups are given high lipid food (HFD), second group as control, 3-6 groups are experimental group, respectively daily gavage 2pmol/g body weight
OCN、ISAP1、ISAP2、ISAP3, continue seven weeks, body weight monitored weekly is once.Experimental result is shown in Fig. 7 A, can be obvious from figure
Find out, with respect to HFD control groups, ISAP2Group occurs in that obvious body weight reduction.
Collect the stool in mice of the 7th week, toasted 3 days at 60 DEG C, it is ensured that drying, then take 1 milligram, with 1ml chloroforms and
The mixed solution immersion of methanol 2 to 1, then crushes excrement with tissue Syrup-homogenizing instrument, supernatant is taken after centrifugation, examined with Roche blood biochemistry instrument
Its triglycerides is surveyed, experimental result is referring to Fig. 7 B.Compared with HFD control groups, ISAP2The content of triglycerides is bright in the excrement of group
Aobvious rise, shows that gavage applies ISAP2Significantly reduce the absorption of triglycerides in enteron aisle.
Embodiment 3
ISAP3Functional study
1st, gavage applies ISAP3Influence to HFD mouse
Using the male wild type C57BL/6 mouse of 6 week old, divide 6 groups, every group 6, the 1st group is given chow diet (ND), 2-
6 groups are given high lipid food (HFD), second group as control, 3-6 groups are experimental group, respectively daily gavage 2pmol/g body weight
OCN、ISAP1、ISAP2、ISAP3, continue seven weeks, body weight monitored weekly is once.Experimental result is shown in Fig. 7 A, can be obvious from figure
Find out, with respect to HFD control groups, ISAP3Group occurs in that obvious body weight reduction.
Collect the stool in mice of the 7th week, toasted 3 days at 60 DEG C, it is ensured that drying, then take 1 milligram, with 1ml chloroforms and
The mixed solution immersion of methanol 2 to 1, then crushes excrement with tissue Syrup-homogenizing instrument, supernatant is taken after centrifugation, examined with Roche blood biochemistry instrument
Its triglycerides is surveyed, experimental result is referring to Fig. 7 B.Compared with HFD control groups, ISAP3The content of triglycerides is bright in the excrement of group
Aobvious rise, shows that gavage applies ISAP3Significantly reduce the absorption of triglycerides in enteron aisle.
Contrast ISAP1、ISAP2、ISAP3Sequence understand：Relative to ISAP1,ISAP2With 4 insertions, 2 are replaced, and
1 missing；Relative to ISAP1, ISAP3With 4 insertions, 3 replacements；Relative to ISAP2, ISAP1With 4 missings, 2
Replace, and 1 insertion；In summary, it is believed that three polypeptides variant each other, presumption can based on these three sequences,
Carry out well known to a person skilled in the art amino acid substitution, insertion and lack, condition is the ability for not making it adjust energetic supersession
Significantly reduce, for example reduction be no more than 40%, 30%, 20%, 10%, referring in accompanying drawing 8 to the same of a variety of different biological sources
Source sequence is compared, wherein SEQ ID NO.2：YLYQWLGAPVPYPDPLEP, SEQ ID NO.4：
YLNNGLGAPAPYPDPLEP, SEQ ID NO.5：YLYQWLGAPVPYPDTLEP, SEQ ID NO.6：
YLYQWLGAPVPYPDPLEP, SEQ ID NO.7：YLDHWLGAPAPYPDPLEP, SEQ ID NO.8：
YLDPGLGAPAPYPDPLEP, SEQ ID NO.9：YLDHGLGAPAPYPDPLEP, SEQ ID NO.10：
YLDQGLGAPAPAPDPLEP, SEQ ID NO.11：Difference very little between YLDSGLGAPVPYPDPLEP is more when comparing two-by-two
Difference therefore can estimate these sequences and have similar biological function no more than the replacement of four amino acid in the case of number,
Should also be as within the scope of the present invention, it is preferable that wherein lack, replace and insertion sum be no more than 4, for example no more than
3、2、1.Other SEQ ID NO.1 have only lacked an amino acid residue compared with SEQ ID NO.18, it is inferred that SEQ ID
NO.1 also has the function similar with SEQ ID NO.18.
Meanwhile, relative to ISAP2, ISAP1、ISAP3Equivalent in ISAP2End has 1 insertion；Showing can be more
Several amino acid residues are added in the end of peptide, and condition is that the ability for not making it adjust energetic supersession is significantly reduced, for example, reduce
No more than 40%, 30%, 20%, 10%, preferably end addition is no more than 5, such as 4,3,2,1 or 0 ammonia
Base acid residue.
Embodiment 4
ISAP4、ISAP5、ISAP6Function
ISAP4、ISAP5、ISAP6Influence of the acute gavage in normal mouse to small intestine fat absorption
Using 6 week old hero wild type C57BL/6 mouse 15, be divided into 5 groups, every group 3, three experimental groups, respectively with
2pmol/g body weight, gavage apply ISAP4、ISAP5、ISAP6；Two groups of controls, daily gavage applies isometric physiological saline, continues
After one week, the 8th, gavage applied ISAP respectively4、ISAP5、ISAP6And 30 minutes after physiological saline, three groups of experimental groups and one
The individual μ L of control group gavage administration of sterile olive oil 200, a control group gavage applies the μ L of physiological saline 200,95% after 50 minutes
CO2Euthanasia mouse simultaneously dissects acquisition small intestine, carries out frozen section to nearly duodenum section jejunum and uses oil red O stain, bush
Element is redyed；Carry out quantitative by ImageJ softwares to Oil red O positive area under double-blind conditions and calculate the area and intestinal villi
The ratio of the gross area.Statistical analysis：Compared with physiological saline+olive oil group.***:p<0.001.N=3..
Experimental result is referring to Fig. 9；Oil red O positive area is quantified and calculated by ImageJ softwares under double-blind conditions
The ratio of the area and the intestinal villi gross area.As can be seen that relative to saline control, ISAP4、ISAP5、ISAP6It is highly effective
Reduce olive oil absorption, it was demonstrated that they and ISAP1、ISAP2、ISAP3It is suitable on biological function, meanwhile, reference picture
8 alignment, it is also seen that SEQ ID NO.12：PVPYPDPLEP and SEQ ID NO.15：SVPSPDPLEP, SEQ ID
NO.13：PYPDPLEP and SEQ ID NO.16：PSPDPLEP similarities in sequence are very high, while between each species
It is also very conservative sequence, thus there should be similar bioactivity, so as to thinks these polypeptides and its variant
Can by orally administer on fat absorption and metabolism produce influence, you can with it is carried out well known to a person skilled in the art
Amino acid substitution, insertion and missing, condition are that the ability for not making it adjust energetic supersession is significantly reduced, and such as reduction is no more than
40%th, 30%, 20%, 10%.Meanwhile, ISAP4、ISAP5、ISAP6It is shorter, therefore cost is lower, stability is more preferable, with use
Make prepare treatment with fat metabolism extremely about disease medicine great potential.
All publications and patents mentioned in this article are incorporated herein by reference in their entirety, such as each single publication
Or patent is specifically and individually shown to be incorporated by reference into.In the case of a conflict, (including herein appointed with the application
What is defined) it is defined.
Equivalent
Although having specifically disclosed some specific embodiments of the present invention herein, description above is to illustrate
Bright property and it is nonrestrictive.For those skilled in the art, by browsing this specification and appended claims, this
Many versions of invention will be apparent.The four corner of the present invention should be by referring to claim, its equivalent
Four corner and this specification and such change are determined.
Sequence table
<110>Shenzhen Xianjin Technology Academe
<120>Adjust polypeptide of energetic supersession and application thereof
<160> 18
<170> PatentIn version 3.3
<210> 1
<211> 14
<212> PRT
<213>Mouse（Mus musculus）
<400> 1
Tyr Leu Gly Ala Ser Val Pro Ser Pro Asp Pro Leu Glu Pro
1 5 10
<210> 2
<211> 18
<212> PRT
<213>People（Homo sapiens）
<400> 2
Tyr Leu Tyr Gln Trp Leu Gly Ala Pro Val Pro Tyr Pro Asp Pro Leu
1 5 10 15
Glu Pro
<210> 3
<211> 19
<212> PRT
<213>Artificial sequence
<220>
<223> ISAP3
<400> 3
Tyr Leu Tyr Gln Trp Leu Gly Ala Pro Val Pro Tyr Pro Asp Pro Leu
1 5 10 15
Glu Pro Arg
<210> 4
<211> 18
<212> PRT
<213>Rat（Rattus norvegicus）
<400> 4
Tyr Leu Asn Asn Gly Leu Gly Ala Pro Ala Pro Tyr Pro Asp Pro Leu
1 5 10 15
Glu Pro
<210> 5
<211> 18
<212> PRT
<213>Bonobo（Pan troglodytes）
<400> 5
Tyr Leu Tyr Gln Trp Leu Gly Ala Pro Val Pro Tyr Pro Asp Thr Leu
1 5 10 15
Glu Pro
<210> 6
<211> 18
<212> PRT
<213>Macaque（Macaca mulatta）
<400> 6
Tyr Leu Tyr Gln Trp Leu Gly Ala Pro Val Pro Tyr Pro Asp Pro Leu
1 5 10 15
Glu Pro
<210> 7
<211> 18
<212> PRT
<213>Ox（Bos taurus）
<400> 7
Tyr Leu Asp His Trp Leu Gly Ala Pro Ala Pro Tyr Pro Asp Pro Leu
1 5 10 15
Glu Pro
<210> 8
<211> 18
<212> PRT
<213>Sheep（Ovis aries）
<400> 8
Tyr Leu Asp Pro Gly Leu Gly Ala Pro Ala Pro Tyr Pro Asp Pro Leu
1 5 10 15
Glu Pro
<210> 9
<211> 18
<212> PRT
<213>Wild boar（Sus scrofa）
<400> 9
Tyr Leu Asp His Gly Leu Gly Ala Pro Ala Pro Tyr Pro Asp Pro Leu
1 5 10 15
Glu Pro
<210> 10
<211> 18
<212> PRT
<213>Naked zokor（Heterocephalus glaber）
<400> 10
Tyr Leu Asp Gln Gly Leu Gly Ala Pro Ala Pro Ala Pro Asp Pro Leu
1 5 10 15
Glu Pro
<210> 11
<211> 18
<212> PRT
<213>Dog（Canis lupus）
<400> 11
Tyr Leu Asp Ser Gly Leu Gly Ala Pro Val Pro Tyr Pro Asp Pro Leu
1 5 10 15
Glu Pro
<210> 12
<211> 10
<212> PRT
<213>People
<400> 12
Pro Val Pro Tyr Pro Asp Pro Leu Glu Pro
1 5 10
<210> 13
<211> 8
<212> PRT
<213>People
<400> 13
Pro Tyr Pro Asp Pro Leu Glu Pro
1 5
<210> 14
<211> 6
<212> PRT
<213>Artificial sequence
<220>
<223> IASP6
<400> 14
Pro Asp Pro Leu Glu Pro
1 5
<210> 15
<211> 10
<212> PRT
<213>Mouse
<400> 15
Ser Val Pro Ser Pro Asp Pro Leu Glu Pro
1 5 10
<210> 16
<211> 8
<212> PRT
<213>Mouse
<400> 16
Pro Ser Pro Asp Pro Leu Glu Pro
1 5
<210> 17
<211> 18
<212> PRT
<213>Artificial sequence
<220>
<223>A kind of ISAP formula
<220>
<221> misc_feature
<222> (3)..(3)
<223>Xaa is Tyr, Asn, Asp or is not present
<220>
<221> misc_feature
<222> (4)..(4)
<223>Xaa is Gln, Asn, His, Pro, Ser or is not present
<220>
<221> misc_feature
<222> (5)..(5)
<223>Xaa is Trp, Gly or is not present
<220>
<221> misc_feature
<222> (6)..(6)
<223>Xaa is Leu or is not present
<220>
<221> misc_feature
<222> (9)..(9)
<223>Xaa is Pro or Ser
<220>
<221> misc_feature
<222> (10)..(10)
<223>Xaa is Ala or Val
<220>
<221> misc_feature
<222> (10)..(10)
<223>Xaa is Tyr or Ser
<220>
<221> misc_feature
<222> (10)..(10)
<223>For Thr or Pro
<400> 21
Tyr Leu Xaa Xaa Xaa Xaa Gly Ala Xaa Xaa Pro Xaa Pro Asp Xaa Leu
1 5 10 15
Glu Pro
<210> 18
<211> 15
<212> PRT
<213>Mouse（Mus musculus）
<400> 18
Tyr Leu Gly Ala Ser Val Pro Ser Pro Asp Pro Leu Glu Pro Thr
1 5 10 15
Claims (19)
1. adjusting the polypeptide of energetic supersession, it is by formula M1-Za-M2Represent, wherein：
M1、M2Be each independently with no more than 5, the polypeptide section of 4,3,2 or 1 amino acid residues or be not present；
ZaFor Tyr-Leu-X1-X2-X3-X4-Gly-Ala-X5-X6-Pro-X7-Pro-Asp-X8- Leu-Glu-Pro, wherein：
X1For Tyr, Asn, Asp or it is not present,
X2For Gln, Asn, His, Pro, Ser or it is not present,
X3For Trp, Gly or it is not present,
X4For Leu or it is not present,
X5For Pro or Ser,
X6For Ala or Val,
X7For Tyr or Ser,
X8For Thr or Pro,
And the ZaOptionally have amino acid substitution, insertion or missing and the amino acid substitution, insertion and lack
Sum is no more than 4.
2. polypeptide according to claim 1, wherein the ZaSelected from one below：
SEQ ID NO.1：YLGASVPSPDPLEP,
SEQ ID NO.2：YLYQWLGAPVPYPDPLEP,
SEQ ID NO.4：YLNNGLGAPAPYPDPLEP,
SEQ ID NO.5：YLYQWLGAPVPYPDTLEP,
SEQ ID NO.6：YLYQWLGAPVPYPDPLEP,
SEQ ID NO.7：YLDHWLGAPAPYPDPLEP,
SEQ ID NO.8：YLDPGLGAPAPYPDPLEP,
SEQ ID NO.9：YLDHGLGAPAPYPDPLEP,
SEQ ID NO.10：YLDQGLGAPAPAPDPLEP,
SEQ ID NO.11：YLDSGLGAPVPYPDPLEP.
3. polypeptide according to claim 2, wherein the M1、M2It is not present.
4. polypeptide according to claim 2, wherein ZaFor YLYQWLGAPVPYPDPLEP, M1It is not present and M2For Arg；Or
Person wherein ZaFor YLGASVPSPDPLEP, M1It is not present and M2For Thr.
5. the polypeptide of energetic supersession is adjusted, its at least six continuous amino acid comprising the polypeptide described in claim 3 and its amino
Sour total number of residues is no more than 18.
6. polypeptide according to claim 5, it includes one below：
SEQ ID NO.12：PVPYPDPLEP,
SEQ ID NO.13：PYPDPLEP,
SEQ ID NO.14：PDPLEP,
SEQ ID NO.15：SVPSPDPLEP,
SEQ ID NO.16：PSPDPLEP.
7. polypeptide according to claim 6, it is one below：
SEQ ID NO.12：PVPYPDPLEP,
SEQ ID NO.13：PYPDPLEP,
SEQ ID NO.14：PDPLEP,
SEQ ID NO.15：SVPSPDPLEP,
SEQ ID NO.16：PSPDPLEP.
8. the officinal salt of the polypeptide any one of claim 1 to 7.
9. polynucleotides, it encodes the polypeptide any one of claim 1 to 7.
10. carrier, it includes the polynucleotides described in claim 9.
11. host cell, it transfects the carrier having the right described in requirement 10 and be able to can produced under conditions of marking protein
Polypeptide any one of raw claim 1 to 7.
12. pharmaceutical composition, polypeptide or claim 8 any one of its claim 1 to 7 comprising therapeutically effective amount
Described officinal salt.
13. officinal salt described in polypeptide or claim 8 or claim 12 institute any one of claim 1 to 7
The pharmaceutical composition stated prepare be used for treat with abnormal energy metabolism about disease medicine in purposes.
14. officinal salt described in polypeptide or claim 8 or claim 12 institute any one of claim 1 to 7
The pharmaceutical composition stated prepare be used for treat with fat metabolism extremely about disease medicine in purposes.
15. the purposes described in claim 14, wherein the disease is reduced for that can benefit from fat absorption, blood fat is reduced, fat
Consumption increase or the disease of Fat Accumulation reduction.
16. the purposes described in claim 15, wherein the disease can be obesity, type ii diabetes, non-alcoholic fatty
Liver, insulin resistance, hypertriglyceridemia, high cholesterol, atherosclerosis, coronary heart disease.
17. officinal salt described in polypeptide or claim 8 or claim 12 institute any one of claim 1 to 7
The pharmaceutical composition stated, it is characterised in that can oral administration.
18. the officinal salt described in polypeptide or claim 8 any one of claim 1 to 7 is being prepared for mitigating
Purposes in the health products of body weight.
19. for the health products lost weight, polypeptide or power any one of its claim 1 to 7 comprising effective dose
Profit requires the officinal salt described in 8.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2018165936A1 (en) * | 2017-03-16 | 2018-09-20 | 深圳先进技术研究院 | Polypeptide for regulating energy metabolism and uses thereof |
| WO2018165933A1 (en) * | 2017-03-16 | 2018-09-20 | 深圳先进技术研究院 | Polypeptide for regulating saccharometabolism and uses thereof |
| CN109959794A (en) * | 2017-12-22 | 2019-07-02 | 深圳先进技术研究院 | A kind of ELISA kit of detection metabolism element |
| CN113993883A (en) * | 2019-08-29 | 2022-01-28 | 渥太华Hdl药物研发公司 | Polypeptide and application thereof |
| CN114288414A (en) * | 2021-12-07 | 2022-04-08 | 深圳先进技术研究院 | Polypeptide crossing blood brain barrier, derivative and application thereof |
| CN114306630A (en) * | 2021-12-07 | 2022-04-12 | 深圳先进技术研究院 | Polypeptide targeting brain tumor, derivative and application thereof |
| WO2023213141A1 (en) * | 2022-05-05 | 2023-11-09 | 中国科学院深圳先进技术研究院 | Ovary-targeting polypeptide, and derivative and use thereof |
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| CN103554249A (en) * | 2013-10-25 | 2014-02-05 | 中国科学院深圳先进技术研究院 | Complete antigen of AG15 polypeptide as well as preparation method and antibody thereof |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018165936A1 (en) * | 2017-03-16 | 2018-09-20 | 深圳先进技术研究院 | Polypeptide for regulating energy metabolism and uses thereof |
| WO2018165933A1 (en) * | 2017-03-16 | 2018-09-20 | 深圳先进技术研究院 | Polypeptide for regulating saccharometabolism and uses thereof |
| US11098083B2 (en) | 2017-03-16 | 2021-08-24 | Shenzhen Institutes Of Advanced Technology | Peptide that regulates fat metabolism and method for regulating fat metabolism |
| CN109959794A (en) * | 2017-12-22 | 2019-07-02 | 深圳先进技术研究院 | A kind of ELISA kit of detection metabolism element |
| CN109959794B (en) * | 2017-12-22 | 2022-05-27 | 深圳先进技术研究院 | ELISA kit for detecting metabolin |
| CN113993883A (en) * | 2019-08-29 | 2022-01-28 | 渥太华Hdl药物研发公司 | Polypeptide and application thereof |
| CN114288414A (en) * | 2021-12-07 | 2022-04-08 | 深圳先进技术研究院 | Polypeptide crossing blood brain barrier, derivative and application thereof |
| CN114306630A (en) * | 2021-12-07 | 2022-04-12 | 深圳先进技术研究院 | Polypeptide targeting brain tumor, derivative and application thereof |
| WO2023213141A1 (en) * | 2022-05-05 | 2023-11-09 | 中国科学院深圳先进技术研究院 | Ovary-targeting polypeptide, and derivative and use thereof |
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| CN107011427B (en) | 2020-06-12 |
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