CN107253985A - The design and its application of one class New-type long-acting incretin peptide - Google Patents
The design and its application of one class New-type long-acting incretin peptide Download PDFInfo
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Abstract
The present invention relates to the design and its synthetic method of a class New-type long-acting incretin peptide.By carrying out transforming the derivatives of Exendin 4 obtained with longer pharmacological action time to Exendin 4, the synthesis of target polypeptides is quickly to realize that crude product is purified by orthogonal Preservation tactics solid phase synthesis process, lyophilized to obtain long-actingization incretin peptide.
Description
Technical field
The present invention relates to the design and its application of a class New-type long-acting incretin peptide in treating diabetes field.
Background technology
Diabetes are the chronic noncommunicable diseases of the third-largest serious threat human health after tumour, angiocardiopathy
Disease.At present, the whole world there are about 300,000,000 diabetics, it is contemplated that will be increased to 500,000,000 by 2025.Clinically controlled using insulin strengthening
The method for the treatment of delays diabetes progression, but insulin injection has the risk of hypoglycemia.Therapeutic effect is by dosage, note
The influence of the factors such as position, injecting pathway is penetrated, and individual difference is larger, it is careless slightly using insulin, just occur serious
Hypoglycemia side effect.
Glucagon-like-peptide-1 (GLP-1) is that a kind of glucose dependency intestines promote blood sugar lowing polypeptide hormone, and GLP-1 is stimulated
Insulin secretion and occur without hypoglycemia, the insulin secretion accelerating characteristic of this glucose dependency, it is to avoid treating diabetes
In the hypoglycemia that is commonly present it is dangerous.Therefore, GLP-1 has wide DEVELOPMENT PROSPECT as a kind of diabetes B medicine.
But natural GLP-1 has many advantages, such as in treatment diabetes, for example, it is in vivo easily by DPP IV
(DPP-IV) fast degradation.DPP-IV can specific recognition GLP-1 8 alanine (Ala) residues of N-terminal, from peptide chain N end
8, end alanine (Ala) place excision dipeptides, makes it be changed into inactive form, its Half-life in vivo only 5min or so.
GLP-1 peptide chain N-terminals are the binding sites with GLP-1 acceptors, if its histidine residues is lost, GLP-1 will be caused to lose completely
Bioactivity.The modification strategy of the extension GLP-1 Half-life in vivo generally used at present is mainly modified 8 so that
GLP-1 can resist the degraded of DPP-IV enzymes, in addition, the amino acid of GLP-1 peptide chains N-terminal 8 and 9, which is exchanged, to be reached
This purpose.Exenatide is the typical short-acting GLP-1 receptor stimulating agents for reducing the metabolism of DPP-IV enzymes.However, due to GLP-1 meetings
Elimination is quickly filtered by kidney, the degraded of resistance DPP-IV enzymes can only extend GLP-1 half-life period to a certain extent.
In this patent, on the basis of the potent activator Exenatide (Exendin-4) of GLP-1 acceptors, using half Guang ammonia
Acid-maleimide conjugation strategy, design has synthesized a class long-actingization incretin peptide.The strategy by the sulfydryl of cysteine with
Michael addition reaction occurs for maleimide easily and efficiently to introduce small molecule group, can avoid in early stage GLP-1 acceptor
In the R＆D process of long-actingization activator, the poor selectivity caused by small molecule group linking arm, reaction are used as using lysine
Inconvenient the problems such as.
In this patent, bicoumarin small molecule and Exenatide are conjugated first, bicoumarin small molecule group have compared with
High seralbumin Percentage bound, can strengthen conjugate and sero-abluminous combination, largely extend compound
Half-life period, and the quick filtration of the kidney of compound and metabolic inactivation, thus the half-life period of such compound and internal drop can be reduced
Sugared action time significantly extends.
In a word, such compound has preferably druggability, can reduce the pain of patient's multiple injection administration, improves disease
People's compliance, is the medicine of great development prospect in diabetes B therapy field.
The content of the invention
The present invention relates to the long-actingization incretin peptide that a class carries ehter bond, it is characterized in that, polypeptid acid sequence is：
The Cys-Gln-Met- of His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser- chemical modifications
Glu-Glu-Glu-Ala-Val- Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-
Gly-Ala-Pro-Pro-Pro-Ser-NH2；Or His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-
Lys-Gln-Met-Glu-Glu-Glu-Ala-Val-Arg-Leu-Phe- Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-
The Cys-NH of Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser- chemical modifications2；Or His-Gly-Glu-Gly-Thr-
The Cys-Gln-Met-Glu-Glu-Glu-Ala-Val- Arg-Leu- of Phe-Thr-Ser-Asp-Leu-Ser- chemical modifications
Tyr-Ile-Gln-Trp-Leu-Lys-Glu-Gly-Gly-Pro-Ser-Ser-Gly-Arg-Pro-Pro-Pro-Ser-NH2；
Or His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met- Glu-Glu-Glu-Ala-
Val-Arg-Leu-Tyr- Ile-Gln-Trp-Leu-Lys-Glu-Gly-Gly-Pro-Ser-Ser-Gly-Arg-Pro-Pro-
The Cys-NH of Pro-Ser- chemical modifications2；
The Cys structures of wherein chemical modification are：
N is derived from 0~20.
The preferred scheme of the present invention, it is characterized in that,
The Cys-Gln-Met- of His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser- chemical modifications
Glu-Glu-Glu-Ala-Val- Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-
Gly-Ala-Pro-Pro-Pro-Ser-NH2；Or His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-
Lys-Gln-Met-Glu-Glu-Glu-Ala-Val-Arg-Leu-Phe- Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-
The Cys-NH of Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser- chemical modifications2；Or His-Gly-Glu-Gly-Thr-
The Cys-Gln-Met-Glu-Glu-Glu-Ala-Val- Arg-Leu- of Phe-Thr-Ser-Asp-Leu-Ser- chemical modifications
Tyr-Ile-Gln-Trp-Leu-Lys-Glu-Gly-Gly-Pro-Ser-Ser-Gly-Arg-Pro-Pro-Pro-Ser-NH2；
Or His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met- Glu-Glu-Glu-Ala-
Val-Arg-Leu-Tyr- Ile-Gln-Trp-Leu-Lys-Glu-Gly-Gly-Pro-Ser-Ser-Gly-Arg-Pro-Pro-
The Cys-NH of Pro-Ser- chemical modifications2；
The Cys structures of wherein chemical modification are：
N is derived from 11~16.
In one embodiment, the present invention relates to the long-actingization incretin peptide with following sequence：
Present invention also offers at least one above-claimed cpd and its medicine of a kind of pharmaceutical composition, including therapeutically effective amount
Acceptable salt on, or pharmaceutically acceptable carrier or diluent.
Invention further provides above-claimed cpd and its pharmaceutically acceptable salt, or pharmaceutically acceptable carrier
Or diluent is preparing the utilization in being used to prevent and treat the medicine of diabetes.
The above-claimed cpd that the present invention is provided has significant hypoglycemic effect, and chemical property is stable, blood sugar reducing function dimension
The time of holding has reached more than 40h, compared with endogenous GLP-1 (2~3min of half-life period) or marketed drug Exenatide (half-life period
2.4h) increase significantly.Simultaneously, it is to avoid the adverse reaction such as local itch caused by pharmacy long-actingization method.
Present invention also offers the preparation method of above-claimed cpd, the present invention is efficiently and rapidly closed using synthesis in solid state strategy
Into obtaining above-mentioned target compound.
The following is internal, the external hypoglycemic pharmacological experimental method and result of the long-actingization incretin peptide being related in the present invention：
(1) the receptor agonist activity experiment of long-actingization incretin peptide
HEK293 cells cotransfection encodes GLP-1R cDNA, and cell line is expressed and utilizes Western Blot detections structure
GLP-1R protein level in the HEK293 cells built is thin with the GLP-R-HEK293 whether investigation establishes stable high expression
Born of the same parents' strain.Receptor agonist activity experiment in, first, by cell kind in 96 orifice plates, after 2h, compound DMSO dissolves, using containing
The culture medium for having 0.1% bovine serum albumin is diluted to different multiples, in the GLP-1R-HEK293 cells for adding cotransfection.It is incubated
After 20min, detected using the ELISA kit of Cisbo companies and compound is calculated after corresponding cAMP values, nonlinear regression
EC50Numerical value.
The receptor agonist activity of the long-actingization incretin peptide of table 1
As shown in table 1, all long-actingization incretin peptides are all retained GLP-1R agonist activity, are drawn with marketed drug profit
Shandong peptide is compared, and compound is all significantly improved to GLP-1R agonist activity.Wherein compound SEQ.ID NO：2 couples of GLP-1R
Agonist activity improve notable, compared with Liraglutide, improve 4.2 times.
(2) the abdominal cavity glucose tolerance test of long-actingization incretin peptide
Normal kunming mice, random packet, every group 8, Mouse feeder is in standardization Animal House.Experiment is prohibited for first 12 hours
Food, is only given drinking-water.Every group of mouse before administration, surveys initial blood glucose value, it is determined as -30min, 25nmol/kg is then injected intraperitoneally
Exenatide or long-actingization incretin peptide.After 30min, 18mmol/kg glucose solution is injected intraperitoneally, it is determined as 0min.0,
15,30,45,60,120min determine blood sugar level with blood glucose meter, detect the hypoglycemic activity of long-actingization incretin peptide.
The abdominal cavity glucose tolerance test result of the long-actingization incretin peptide of table 2
Results are expressed as mean ± SD, * P ＜ 0.05, * * P ＜ 0.01, * * * P ＜ 0.001vs
saline.
As shown in table 2, it is hypoglycemic test result indicates that, the long-actingization incretin peptide that is related in the present invention, blood sugar decreasing effect with
Exenatide is suitable.
(3) the next day hypoglycemic experiment of long-actingization incretin peptide
After abdominal cavity glucose tolerance test terminates, immediately normal diet drinking-water 10h, then fasting 12h, carries out mouse again
Abdominal cavity glucose tolerance test.Each group mouse peritoneal injection 18mmol/kg glucose solution, the injectable dextrose monohydrate time is set to 0
Min, blood sugar level is determined in 0,15,30,45,60 and 120min with blood glucose meter.
The next day hypoglycemic effect of the long-actingization incretin peptide of table 3
Results are expressed as mean ± SD, * P ＜ 0.05, * * P ＜ 0.01, * * * P ＜ 0.001vs
saline.
As shown in table 3, hypoglycemic experimental result is shown, the long-actingization incretin peptide being related in the present invention is metabolized 24h in vivo
Still there is reduction blood glucose effect afterwards, and Exenatide loses activity already.Illustrate the long-actingization incretin peptide obtained after modification
Biological half-life all significantly extends, and has reached more than 30h.
(4) the stabilizing blood sugar experiment of long-actingization incretin peptide
The blood glucose of the diabetic mice of STZ inductions is determined, mouse of the selection blood glucose numerical value higher than 20mmol/L is carried out
Random packet, every group 6, mouse free choice feeding during experiment.Exenatide or Liraglutide is injected intraperitoneally in positive controls,
Negative control group intraperitoneal injection of saline, administration group injects long-actingization incretin peptide respectively.0h gives compound, respectively 0,
1st, 2,3,4,6,8,12,24,36 and 48h determines blood sugar level using blood glucose meter.After evaluation index is intraperitoneal injection compound,
Mouse blood sugar numerical value is less than 8.35mmol/L time.
The stabilizing blood sugar experiment of the long-actingization incretin peptide of table 4
Results are expressed as mean ± SD, * P ＜ 0.05, * * P ＜ 0.01, * * * P ＜ 0.001vs
saline.
From table 4, SEQ.ID NO：The time of 1 stabilizing blood sugar up to 48.4h, far above the 11.2h of Liraglutide.Table
It is bright under identical experiment condition, compared with marketed drug Liraglutide and Exenatide, the present invention in be related to long-actingization drop
Glycopeptide can reach more excellent long-actingization hypoglycemic effect, the potentiality with the hypoglycemic medicine for being developed into being administered once for 2 days.
The advantage of the invention is that：
1., propose a kind of long-actingization incretin peptide, with stronger GLP-1 receptor agonist activities, with marketed drug Ai Sai
That peptide is compared with Liraglutide, can reach more excellent long-actingization hypoglycemic effect.
2nd, a kind of long-actingization incretin peptide proposed, with excellent long-actingization hypoglycemic effect, the stabilizing blood sugar time is up to
More than 48.4h, the Liraglutide being relatively administered once a day compared to notable extension, such long-actingization incretin peptide have it is preferable into
The property of medicine, can reduce the pain of patient's multiple dosing, be the medicine of great development prospect in existing new chemical entities.
3rd, a kind of long-actingization incretin peptide proposed, high income, synthesis cycle are short, purifying crude is easy, production cost is low, easily
In industrial automation production.
In summary, the long-actingization incretin peptide that the present invention is provided, structure is brand-new, than Exenatide prototype or endogenous GLP-
1 more stablizes, and the hypoglycemic effect time than marketed drug Liraglutide is longer, is suitable as the new of Remedies for diabetes
Type active component, new breakthrough is brought to treating diabetes field.
Embodiment
Following abbreviation is used in this specification：
Ala：Alanine；Arg：Arginine；Asn：Asparagine；Asp：Asparatate；DCM：Dichloromethane； DIC：
N, N '-DIC；DIEA：N, N '-diisopropylethylamine；DMAP：DMAP； DMF：Dimethyl
Formamide；DMSO：Dimethyl sulfoxide；EDC.HCl：1- ethyls-(3- dimethylaminopropyls) carbodiimide hydrochloride；ESI-MS：
Electrospray ionization mass spectrum；Et3N：Triethylamine；Fmoc：N-9- fluorenylmethyloxycarbonyls；Gln：Glutamine；Glu：Glutamic acid；Gly：Sweet ammonia
Acid；HBTU：BTA-N, N, N ', N '-tetramethylurea hexafluorophosphoric acid ester；His：Histidine；HOBt：1- hydroxyls-benzo
Triazole；HPLC：High performance liquid chromatography；Ile：Isoleucine；Leu：Leucine； Lys：Lysine；Met：Methionine；
NMP：1-METHYLPYRROLIDONE；Phe：Phenylalanine；Pro：Proline； Ser：Serine；Thr：Threonine；Trp：Tryptophan；
Tyr：Tyrosine；Val：Valine.
The present invention is illustrated by the following example, but these embodiments do not do the solution of any limitation present invention
Release.
Embodiment 1
Synthesis in solid state.
1st, cysteine changes the synthesis of structure polypeptide peptide chain
1.1st, resin is swelled
Fmoc-Rink amide-MBHA Resin 50mg (substitution value 0.4mmol/g) are weighed, 30 are swelled through DCM 7mL
Min, suction filtration removes DCM, then is swelled 30min with NMP 10mL, finally uses NMP, DCM, NMP 7mL to rinse well respectively.
1.2nd, the removing of Fmoc protection groups
The resin being swelled is put into reactor, 25% piperidines/NMP (V/V) solution 7mL of the HOBt containing 0.1M is added,
1min is reacted, solution is filtered off after terminating；25% piperidines/NMP (V/V) solution 7mL of the HOBt containing 0.1M is added, is reacted
4min, solution is filtered off after terminating, and uses NMP washes cleans.Obtain sloughing the resin of the Fmoc protection groups initially connected.
1.3rd, Fmoc-Ser (tBu)-Rink amide-MBHA Resin synthesis
By Fmoc-Ser (tBu)-OH (15.3mg, 0.04mmol), HBTU (15.1mg, 0.04mmol), HOBt (5.4mg,
0.04mmol) it is dissolved in DIPEA (13.9 μ L, 0.08mmol) in NMP 10mL, then this solution is added what step 1.1 was obtained
In resin, 7min is reacted, reaction solution is filtered off after terminating, resin is washed 3 times with each 7mL of DCM and NMP.
1.4th, the detection of coupling efficiency
Take a small amount of resin particle DMF to wash, be put into the bromophenol blue solution that 3 drops 1% are added in transparent vials, normal temperature shaking 3
Minute, the aobvious blueness of resin is the positive, transparent for feminine gender.If feminine gender can just enter next coupling cycles.
1.5th, the extension of peptide chain
According to the sequence of peptide chain, repeat above-mentioned deprotection and be sequentially connected corresponding amino acid the step of coupling, successively
Corresponding amino acid is finished up to peptide chain is synthesized in connection, obtains being connected with the resin of polypeptide chain.
1.6th, on resin polypeptide cracking
The resin obtained above for being connected with polypeptide chain is put into reaction bulb, decomposition agent Reagent K (TFA/ benzene first is added
Thioether/water/phenol/EDT, 82.5: 5: 5: 5: 2.5, V/V) 10mL, 30min is first shaken at 0 DEG C, then react at normal temperatures
3h.Reaction terminates rear suction filtration, plus a small amount of TFA and DCM is washed three times, merging filtrate.Filtrate is added in substantial amounts of ice ether and analysed
Go out white flock precipitate, refrigerated centrifuge obtains the crude product of target polypeptides.The crude product 37.1mg of compound is finally given, yield is
89.2%.
2nd, the synthesis of chemical modification base
The synthesis of 12- maleic amide lauric acid/dodecanoic acids
12-aminolauric acid (0.86g, 4mmol) and maleic anhydride (0.47g, 4.8mmol) are dissolved in glacial acetic acid, 120
DEG C heating response 6h, after lamellae detection reaction completely, is cooled to room temperature by reaction solution, is poured into water, be extracted with ethyl acetate
Three times (3 × 20mL), merges upper layer of extraction liquid, and extract is washed 3 times using saturated common salt, and anhydrous Na 2SO4 is dried overnight.Will
Extract is concentrated under reduced pressure, and obtained crude product purifies to obtain faint yellow sterling 0.88g, yield 75%, mp 91-92 DEG C through column chromatography.
1H-NMR(DMSO-d6, 300MHz)：δppm：12.42 (s, 1H ,-COOH), 7.50 (s, 2H ,-COCH=CHCO-),
3.88 (t, 2H, J=7.0Hz ,-NCH2-), 2.68 (t, J=7.3Hz, 2H ,-CH2COOH), 2.00-1.96 (m, 4H ,-
NCH2CH2(CH2)7CH2), 1.73 (s, 14H ,-NCH2CH2(CH2)7CH2).MS (ESI, m/z)：294.1[M+H]+.
The synthesis of 3,3 '-(4- nitros benzylidene)-two -4 hydroxy coumarins
Paranitrobenzaldehyde (3.02g, 0.02mol) is weighed, is dissolved with absolute ethyl alcohol 35ml；Add 4 hydroxy coumarin
(6.6g, 0.041mol), adding absolute ethyl alcohol 15ml makes dissolving complete.80 DEG C of reaction 4h, are filtered, filter cake hot ethanol while hot
30ml is washed 3 times, obtains product 8.2g, yield 90.0%, 227 DEG C of mp.
1H-NMR(CDCl3, 300MHz) and δ ppm：6.13 (s, H ,-CH-), 7.43 (m, 8H, Ar-H), 7.68 (m, 2H, Ar-
H), 8.18 (m, 2H, Ar-H) .MS (ESI, m/z)：456.0[M+H]+.
The synthesis of 3,3 '-(4- amino benzylidene)-two -4 hydroxy coumarins
3,3 '-(4- nitros benzylidene)-two -4 hydroxy coumarins (1.14g, 0.0025mol) are weighed, acetic acid 30ml is used
It is allowed to suspend, adds 5%Pd/C 0.3g, stirring, hydrogen threeway is evacuated 3 times, bottleneck applies vaseline, normal temperature hydrogenation was reacted
Night, suction filtration, filtrate boils off partial solvent, and acetone recrystallization obtains product 0.8g, yield 75.1%, 220 DEG C of mp.
1H-NMR(DMSO-d6, 300MHz) and δ ppm：6.27 (s, H ,-CH-), 7.23 (m, 8H, Ar-H), 7.49 (m, 2H,
Ar-H), 7.8l (m, 2H, Ar-H) .MS (ESI, m/z)：426.0[M+H]+.
The synthesis of 3,3 '-(4- (12- dimaleoyl imino lauroyls amido) benzylidene)-two -4 hydroxy coumarins
12- maleic amides lauric acid/dodecanoic acid (294.1mg, 1mmol) is dissolved in tetrahydrofuran, plus DIC (17 μ L, 1.1mmol) and
HOBt (148.5mg, 1.1mmol), is stirred at room temperature 30min and carrys out activated carboxyl, above-mentioned solution then is slowly dropped into 3,3 '-(4-
Amino benzylidene)-two -4 hydroxy coumarin and DIPEA (17.4 μ L, 0.1mmol) tetrahydrofuran solution, be stirred at room temperature
Night.Reaction solution is poured into water and is extracted with ethyl acetate three times after terminating by lamellae detection reaction, and combining extraction liquid is used respectively
K2CO3, HCl 1M, saturated common salt washing three times.Extract adds anhydrous Na2SO4It is dried overnight, is concentrated under reduced pressure, obtains crude product, post
Chromatographic purifying obtains sterling, yield 69%, mp 204-206 DEG C.
1H-NMR(DMSO-d6, 300MHz)：δppm：10.17 (s, 1H ,-CONH-), 8.31 (d, J=7.8Hz, 2H, Ar-
H), 8.00 (t, J=7.2Hz, 2H, Ar-H), 7.84 (d, J=8.0Hz, 2H, Ar-H), 7.76-7.72 (m, 6H, Ar-H),
7.49 (s, 2H ,-COCH=CHCO-), 6.70 (s, 1H ,-CH-), 3.87 (t, J=7.0Hz, 2H ,-NCH2-), 2.74 (t, J=
7.2Hz, 2H ,-COCH2-), 2.05-1.97 (m, 4H ,-NCH2CH2(CH2)7CH2-), 1.70 (s, 14H ,-NCH2CH2(CH2)7CH2-) MS (ESI, m/z)：703.1[M+H]+.
3rd, the Cys of chemical modification12The synthesis and purifying of-Exenatide conjugate
3 that upper step is obtained, 3 '-(4- (12- dimaleoyl imino lauroyls amido) benzylidene)-two -4- hydroxyls are fragrant
Legumin dissolves the solution for being made into about 10mg/mL with DMSO, and the structure Exenatide polypeptide analog that changes that Cys is replaced also is dissolved in
DMSO, stirring reaction at room temperature after both mix adds 20 μ l DIEPA to accelerate reaction, uses LC-MS monitoring reaction feelings
Condition.Chromatographic condition is：C18 reversed-phase columns (1.7 μm of 2.1 × 50mm, Waters)；Mobile phase A：0.1% formic acid/water (V/V), stream
Dynamic phase B：0.1% formic acid/acetonitrile (V/V), eluent gradient：Mobile phase B 10%~90%, 2min, B 90%~90%,
3min；Flow velocity is 0.3ml/min；Ultraviolet detection wavelength is 214nm.After reaction terminates, reaction solution, which is used, contains 1% trifluoro second
High speed centrifugation and using after 0.45 μm of filtering with microporous membrane after the dilution in acetonitrile of acid, is purified using preparative liquid chromatography,
Chromatographic condition is：C18 reversed-phase columns (320mm × 28mm, 5 μm)；Mobile phase A：0.1% trifluoracetic acid/water (V/V), Mobile phase B：
0.1% trifluoracetic acid/acetonitrile (V/V)；Eluent gradient：Mobile phase B 40%~80%, 30min；80%~85%, 10min；
85%~95%, 10min；95%~40%, 10min；Flow velocity is 5ml/min, and Detection wavelength is 214nm.Solution is collected, is subtracted
Pressure concentration removes acetonitrile, lyophilized to produce sterling.Theoretical relative molecular mass is 4900.0.ESI-MS m/z：Calcd. [M+3H
]3+1634.3, [M+4H]4+1226.0；Found[M+3H]3+1634.0, [M+4H]4+1226.1。
Embodiment 2
Synthetic method be the same as Example 1, theoretical relative molecular mass is 5028.2.ESI-MS m/z：Calcd.[M+3H]3+
1677.1, [M+4H]4+1258.1；Found[M+3H]3+1677.8, [M+4H]4+1258.5。
Embodiment 3
Synthetic method be the same as Example 1, theoretical relative molecular mass is 5015.1.ESI-MS m/z：Calcd.[M+3H]3+
1672.7, [M+4H]4+1254.8；Found[M+3H]3+1672.2, [M+4H]4+1254.3。
Embodiment 4
Synthetic method be the same as Example 1, theoretical relative molecular mass is 5143.3.ESI-MS m/z：Calcd.[M+3H]3+
1715.4, [M+4H]4+1286.8；Found[M+3H]3+1715.4, [M+4H]4+1286.2。
Claims (9)
1. the present invention relates to a class New-type long-acting incretin peptide, it is characterized in that, polypeptid acid sequence is：
The Cys-Gln-Met-Glu- of His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser- chemical modifications
Glu-Glu-Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-
Ala-Pro-Pro-Pro-Ser-NH2；Or His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-
Gln-Met-Glu-Glu-Glu-Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-
The Cys-NH of Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser- chemical modifications2；Or His-Gly-Glu-Gly-Thr-Phe-
The Cys-Gln-Met-Glu-Glu-Glu-Ala-Val-Arg-Leu-Tyr- of Thr-Ser-Asp-Leu-Ser- chemical modifications
Ile-Gln-Trp-Leu-Lys-Glu-Gly-Gly-Pro-Ser-Ser-Gly-Arg-Pro-Pro-Pro-Ser-NH2；Or
His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Glu-Ala-Va1-
Arg-Leu-Tyr-Ile-Gln-Trp-Leu-Lys-Glu-Gly-Gly-Pro-Ser-Ser-Gly-Arg-Pro-Pro-Pro-
The Cys-NH of Ser- chemical modifications2；
The Cys structures of wherein chemical modification are：
N is derived from 0~20.
2. compound according to claim 1, it is characterized in that, polypeptid acid sequence is：
The Cys-Gln-Met-Glu- of His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser- chemical modifications
Glu-Glu-Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-
Ala-Pro-Pro-Pro-Ser-NH2；Or His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-
Gln-Met-Glu-Glu-Glu-Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-
The Cys-NH of Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser- chemical modifications2；Or His-Gly-Glu-Gly-Thr-Phe-
The Cys-Gln-Met-Glu-Glu-Glu-Ala-Val-Arg-Leu-Tyr- of Thr-Ser-Asp-Leu-Ser- chemical modifications
Ile-Gln-Trp-Leu-Lys-Glu-Gly-Gly-Pro-Ser-Ser-Gly-Arg-Pro-Pro-Pro-Ser-NH2；Or
His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Glu-Ala-Val-
Arg-Leu-Tyr-Ile-Gln-Trp-Leu-Lys-Glu-Gly-Gly-Pro-Ser-Ser-Gly-Arg-Pro-Pro-Pro-
The Cys-NH of Ser- chemical modifications2；
The Cys structures of wherein chemical modification are：
N is derived from 11~16.
3. the compound according to any one in claim 1 to 2, is selected from：
4. a kind of pharmaceutically acceptable salt, described medicine according to prepared by the compound of any one in claims 1 to 3
Acceptable salt is and hydrochloric acid on, hydrobromic acid, sulfuric acid, nitric acid or phosphoric acid；Butanedioic acid, maleic acid, acetic acid, fumaric acid, lemon
The salt of lemon acid, citric acid, tartaric acid, benzoic acid, benzene sulfonic acid, methanesulfonic acid or naphthalene sulfonic acids formation.
5. the medicament according to prepared by the compound of any one in claims 1 to 3, described medicament is any medicine
The upper described tablet of agent, capsule, elixir, syrup, lozenge, inhalant, spray, injection, film, patch, powder, particle
Agent, block agent, emulsion, suppository, compound preparation.
6. according to the compound of any one in claims 1 to 3, answering in the medicine for treating or preventing diabetes is prepared
With.
7. a kind of pharmaceutically acceptable salt according to prepared by the compound of any one in claims 1 to 3, is controlled in preparation
Application in the medicine for the treatment of or prevention diabetes.
8. the medicament according to prepared by the compound of any one in claims 1 to 3, diabetes are treated or prevented preparing
Medicine in application.
9. according to the preparation method of the compound of any one in claims 1 to 3, including biological expression, liquid phase synthesis and
Solid phase synthesis preparation method thereof.
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| CN108948213A (en) * | 2018-07-25 | 2018-12-07 | 中国药科大学 | Long-actingization oxyntomodulin (OXM) hybrid peptide, preparation method and its purposes as drug |
| CN108948212A (en) * | 2018-07-25 | 2018-12-07 | 中国药科大学 | Long-actingization oxyntomodulin (OXM) hybrid peptide and the preparation method and application thereof |
| CN109942695A (en) * | 2017-12-21 | 2019-06-28 | 中国药科大学 | Long-actingization Exenatide (Exendin-4) analog and its application |
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