CN101830969A - Balsam pear hypoglycemic MC-JJ0104 polypeptide analogue by microwave-accelerated solid-phase synthesis and application thereof - Google Patents

Balsam pear hypoglycemic MC-JJ0104 polypeptide analogue by microwave-accelerated solid-phase synthesis and application thereof Download PDF

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CN101830969A
CN101830969A CN201010178102A CN201010178102A CN101830969A CN 101830969 A CN101830969 A CN 101830969A CN 201010178102 A CN201010178102 A CN 201010178102A CN 201010178102 A CN201010178102 A CN 201010178102A CN 101830969 A CN101830969 A CN 101830969A
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tyr
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黄文龙
金晶
钱海
张惠斌
王敬杰
陈巍
朱孝云
华杰
杜阔
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China Pharmaceutical University
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China Pharmaceutical University
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Abstract

The invention relates to a novel high-efficiency hypoglycemic balsam pear MC-JJ0104 polypeptide analogue and a microwave-accelerated solid-phase synthesis method. The MC-JJ0104 polypeptide analogue with longer pharmacological action time is obtained by modifying the 3, 4, 8 or 9 locus of a natural balsam pear polypeptide. The chemical synthesis of the polypeptide analogue is efficiently and quickly realized by the microwave-accelerated solid-phase synthesis method. The crude product is purified by a preparative scale high-efficiency liquid phase, and the MC-JJ0104 polypeptide analogue is finally obtained through freeze-drying.

Description

Balsam pear hypoglycemic MC-JJ 0104 polypeptide analogue by microwave-accelerated solid-phase synthesis and application thereof
Technical field
The present invention relates to MC-JJ0104 polypeptide analog and microwave thereof and promote solid phase synthesis process.
Background technology
Diabetes are important diseases of current harm humans health, and the whole world has the people of 5%-7% to be subjected to diabetes mellitus approximately at present, is considered to after cardiovascular and cerebrovascular diseases, malignant tumour, cause human wounded or disabled, dead the third-largest killer.
Diabetes are divided into I type and II type, and type i diabetes is a kind of autoimmune disease, and patient's survival needs insulin injection.Type ii diabetes accounts for about 95% of numerous diabetic subject's sums.The reason of type ii diabetes is an islet cells metabolism generation obstacle, hypoinsulinism, perhaps producing insulin resistant, is to be main companion's insulin secretion relative deficiency with the insulin resistant, serves as the metabolic disease that main companion's insulin resistant causes with the insulin secretion relative deficiency perhaps.Hypoinsulinism perhaps produces insulin resistant and all can cause in the blood glucose excessive, and when blood-sugar content surpasses the kidney presentation time, glucose is just transferred in the urine and discharged, thereby produces dextrosuria.
The several drugs of clinical application at present all can not be cured diabetes, mainly is by different mechanism of action, reduces patient's glucose level.The diabetic subject must take medicine throughout one's life, and medicine itself is threatening diabetics's health equally to the toxic side effect of human body.In China, use at present medicine can controlling blood sugar to reach arm's length standard, 20% (the Zou Dajin that is less than diabetic subject's sum, the chief culprit of diabetes B---insulin resistant, Popular Medicine, 2002, (6): 17), directly caused many diabetic subjects' complication to take place early, progress is fast.Obviously, except screening more safely and effectively the newtype drug, seeking other the effective glucose level control of auxiliary treatment means, is crucial for the health level that improves the diabetics equally.
Balsam pear (Momordica charantia) is that a kind of mankind have the edible historical vegetables of long-term safety.Recent two decades comes, people have found many compositions with hypoglycemic activity in the fruit of balsam pear, the extract of some Fructus Momordicae charantiaes, perhaps directly with the tea drink of making after the Fructus Momordicae charantiae drying, recommended the diabetics and used, some balsam pear composition has been applied the patent as the treatment diabetes medicament.But, from scientific and technical literature, introduce and existing patent or product, mainly be based on in the many blood-sugar decreasing actives of balsam pear to certain understanding of principal component, do not recognize, have in the balsam pear composition of hypoglycemic activity known just have multiple, and chemical property has nothing in common with each other, and in addition, it is unknown also having some to have hypoglycemic activeconstituents in the balsam pear.So, existing document, patent or product, its limitation also is tangible.
Can develop the different all kinds of hypoglycemic activity things of physico-chemical property in the cover energy full use balsam pear, and the active component content height, so, also be very necessary for fully and reasonably utilizing and develop the balsam pear resource.
Polypeptide can obtain by the method for engineered method or chemosynthesis.Engineered method has superiority than chemical process obtaining on long peptide (amino-acid residue length is greater than 50) or the protein, but the chemical synthesis process of polypeptide is especially after the solid phase synthesis strategy occurs, handiness, diversity and the high efficiency etc. that have gene engineering method to hardly match on less than 40 polypeptide or little peptide in preparation amino-acid residue length.Merrifield in 1963 found and have developed the method for solid-phase synthetic peptide.Solid-phase polypeptide is synthetic generally two kinds of strategies: i.e. Boc/Bzl orthogonally protect solid phase synthesis strategy and Fmoc/tBu orthogonally protect solid phase synthesis strategy, the solid-phase synthesis that Merrifield foundes is Boc/Bzl orthogonally protect solid phase synthesis strategy.But, have some shortcomings in the Boc/Bzl orthogonally protect solid phase synthesis strategy, many as side reaction, condition is harsh and in the process of prolongation peptide chain polypeptide chain can lose from solid phase etc.The Fmoc/tBu orthogonally protect solid phase synthesis strategy of Chu Xianing reaction conditions gentleness many then subsequently.After microwave technology applies in the solid phase synthesis of polypeptide in recent years, make the synthetic technology of polypeptide produce a leap.Microwave promotes chemical reaction to be because it makes polar molecule fast rotational in microwave field, makes fast 10 to 1000 times of the more conventional heating means of some speed of reaction, and productive rate improves greatly.People's reported first such as Gedye in 1986 with microwave application in organic synthesis, microwave can promote many chemical reactions, as the Diels-Alder reaction, saponification reaction etc. can be accelerated speed of reaction significantly and improve yield.In 1992, Hui-ming Yu etc. reported at first with microwave application that in polypeptide solid phase synthesis field they have finished the synthetic of acyl carrier protein segment decapeptide (acyl carrier protein, ACP 65～74).
How synthetic efficiently long peptide remain a challenge in the world, because polypeptide is along with the increase of peptide chain, its synthetic difficulty is multiplied.With the synthetic long peptide of traditional solid phase method, the more purification difficult of the often very low impurity of its yield, thereby say from practical angle and to have lost the synthetic meaning.Secondly synthesis cycle is long, and 10 peptides nearly time in a week of general needs just can not finished synthesis cycle under the centre does not encounter difficulties the situation of peptide preface, if difficult peptide preface, often just can't synthesize and obtains target polypeptides.And use microwave to promote solid-phase synthetic peptide, can overcome difficulties the peptide preface, the polypeptide that synthetic traditional solid phase method can't obtain, and generated time shortens greatly, and significantly improve polypeptide crude product purity and yield, make that the purifying of product is convenient greatly, as long as just can obtain the purity high product through a preparation HPLC; Can carry out pointed decoration with D type amino acid and alpha-non-natural amino acid etc. to polypeptide very easily in addition, remove to seek active higher, polypeptide that biological half-life is longer, this especially genetic engineering technique can't accomplish.Therefore, our employing this kind method synthetic MC-JJ0104 serial analogs that obtains rapidly and efficiently in balsam pear hypoglycemic (MC-JJ0104) polypeptide analog synthetic.
Summary of the invention
It is to obtain polypeptide to modify and obtain the higher MC-JJ0104 polypeptide analog of hypoglycemic activity from the medicinal and edible plant balsam pear of tool hypoglycemic activity that the present invention has two first purposes of purpose.
Second purpose of the present invention provided the method for preparing solid phase of MC-JJ0104 polypeptide derivative, and the present invention adopts microwave to promote Fmoc/tBu orthogonally protect solid phase synthesis strategy efficiently to synthesize apace and obtains serial MC-JJ0104 polypeptide analog.
It is characterized in that: adopt microwave to promote Fmoc/tBu orthogonally protect solid phase synthesis strategy, earlier syntheticly on solid phase carrier obtain being loaded with first Fmoc and protect amino acid whose resin, ninhydrin method detects sloughs the resin that the Fmoc protecting group obtains being loaded with first amino-acid residue after negative; Enter next coupling circulation again; repeat the step of coupling and deprotection with different protection amino acid according to corresponding peptide preface; prolong required aminoacid sequence successively, the synthetic resin that obtains being loaded with corresponding polypeptide cuts down polypeptide with cracking agent at last and obtains the polypeptide crude product from resin.Crude product is through the preparation scale high-efficient liquid phase chromatogram purification, and freeze-drying gets the pure product of polypeptide.
Be loaded with the preparation method that first Fmoc protects amino acid whose resin; it is characterized in that by obtaining with the solid phase carrier coupling again after the activated dose of activation earlier of Fmoc protection amino acid under the microwave irradiation condition; add 1-hydroxyl-benzotriazole (HOBT) or derivatives thereof in the reaction and suppress racemization, and use the acidity of organic bases neutralization reaction.Being loaded with first Fmoc, to protect the employed solid phase carrier of amino acid whose resin be Rink resin, Wang resin or 2-chlorine trityl chloride resin.Activator is dicyclohexylcarbodiimide (DCC), N, N '-DIC (DIC), N, N " carbonyl dimidazoles (CDI); 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC.HCl); 2-(7-azo benzotriazole)-N; N; N '; N '-tetramethyl-urea phosphofluoric acid ester (HATU), benzotriazole-N, N, N ', N '-tetramethyl-urea phosphofluoric acid ester (HBTU) or 1-hydroxyl-benzotriazole (HOBT) derivative that uses are N-hydroxy-succinamide (HOSU), 1-hydroxyl-7-azo benzotriazole (HOAT) or 3-hydroxyl-1,2,3-phentriazine-4 (3H)-ketone (HOOBT).Organic bases is triethylamine (TEA), N-methylmorpholine (NMM) or diisopropylethylamine (DIEA); Microwave promotion condition is: microwave frequency 2450MHz, temperature of reaction: 20～100 ℃, the reaction times is 5～15min.
Removing of Fmoc protecting group is to contain 0.1molL by use -1Hexahydropyridine solution reaction under microwave promotes of 1-hydroxyl-benzotriazole (HOBT), selecting dimethyl formamide (DM F), methyl-sulphoxide (DMSO) or N-Methyl pyrrolidone (NMP) for use is reaction solvent.Microwave promotion condition is: microwave frequency 2450MHz, temperature of reaction: 20～100 ℃, the reaction times is 1～10min.
The invention has the advantages that:
1. a kind of MC-JJ0104 polypeptide analog of Ti Chuing can improve the stability of MC-JJ0104 on the basis that keeps hypoglycemic activity, prolongs action time.
2. microwave promotes solid phase synthesis MC-JJ0104 polypeptide analog to improve coupled reaction speed greatly, and amino acid of the abundant coupling of conventional solid phase synthesis process goes to resin, often needs do not wait by 20 hours in 2 hours, even longer.Microwave promotes then average to need about 10 minutes; Conventional solid phase synthesis process takes off the Fmoc protecting group, often needs do not wait by 1 hour in 30 minutes, and microwave promotes then on average only to need about 5 minutes, and this has improved polypeptide synthetic efficient greatly, has shortened synthesis cycle.
3. microwave promotes the purity of the synthetic crude product that obtains of solid phase synthesis MC-JJ0104 polypeptide analog greater than 60%, more conventional solid phase synthesis process improves greatly, this has made things convenient for follow-up purifying work, only need be through once preparing the liquid phase purifying, and freeze-drying can obtain the pure product of target.
4. microwave promotes the synthetic MC-JJ0104 polypeptide analog of solid phase method, and its cost is low because coupling efficiency is higher, required protection amino acid is average only need 2 times excessive, more conventional solid phase synthesis process needs 4 to 5 times excessively greatly to reduce.
5. microwave promotes solid phase synthesis MC-JJ0104 polypeptide analog method easily to be automated, to change on a large scale, and this makes it be more suitable for suitability for industrialized production.
Therefore promote the MC-JJ0104 polypeptide analog that solid phase synthesis technique prepares with microwave provided by the invention, the yield height, synthesis cycle is short, purifying crude is easy, and production cost is low, be easy to industrial automation production.The MC-JJ0104 polypeptide analog for preparing, more stable than natural MC-JJ0104, be suitable as the activeconstituents for the treatment of diabetes medicament.
Embodiment
Adopt following abbreviation in this specification:
Et 3N: triethylamine; The NMM:N-methylmorpholine; DIEA:N, N '-diisopropylethylamine; DMF: dimethyl formamide; DM SO: methyl-sulphoxide; DCM: methylene dichloride; The Fmoc:N-9 fluorenylmethyloxycarbonyl; DIC:N, N '-DIC; CDI:N, N '-carbonyl dimidazoles; The DMAP:4-Dimethylamino pyridine; The HOSU:N-N-Hydroxysuccinimide; EDC.HCl:1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride; HATU:2-(7-azo benzotriazole)-N, N, N ', N '-tetramethyl-urea phosphofluoric acid ester; HBTU: benzotriazole-N, N, N ', N '-tetramethyl-urea phosphofluoric acid ester; HCTU:6-chlorobenzene and triazole-1,1,3,3-tetramethyl-urea phosphofluoric acid ester; HOAT:1-hydroxyl-7-azo benzotriazole; HOBT:1-hydroxyl-benzotriazole; PyBOP: phosphofluoric acid benzotriazole-1-base-oxygen base tripyrrole alkyl phosphorus; HPLC: high performance liquid chromatography; ESI-MS: electrospray ionization mass spectrum; Gly: glycine; Ser: Serine; Ala: L-Ala; Thr: Threonine; Val: Xie Ansuan; Ile: Isoleucine; Leu: leucine; Tyr: tyrosine; Phe: phenylalanine; His: Histidine; Pro: proline(Pro); Asp: aspartic acid; Met: methionine(Met); Glu: L-glutamic acid; Trp: tryptophane; Lys: Methionin; Arg: arginine.Asn: l-asparagine; Gln: glutamine.
The present invention describes by following implementation column, but these embodiment do not do any restriction explanation of the present invention.
Embodiment 1
The microwave of Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Ala-Ser (SEQ.ID NO:2) promotes solid phase synthesis
(1) swelling of resin
Take by weighing Fmoc-Ala Resin 50mg (replacement amount 0.4mmol/g), through 7mL DCM swelling 30min, suction filtration removes DCM, uses 10mL NMP swelling 30min again, uses NMP at last respectively, DCM, and NMP 7mL rinses well.
(2) microwave promotes removing of Fmoc protecting group
The resin that swelling is good is put into reactor, adds 25% piperidines/NMP (V/V) solution that 7mL contains 0.1M HOBT, reacts 1min in microwave reactor, microwave power is 15W, temperature of reaction is controlled in 50 ℃, uses the cooling of air compressor pressurized air, and reaction finishes back elimination solution; Add 25% piperidines/NMP (V/V) solution that 7mL contains 0.1M HOBT again and react 4min again in microwave reactor, microwave power is 25W, and temperature of reaction is controlled at 50 ℃, uses the cooling of air compressor pressurized air.Reaction finishes back elimination solution, uses the NMP washes clean.Obtain sloughing the resin of the Fmoc protecting group of initial connection.
(3) microwave promotes synthesizing of Fmoc-Ala-rink amide-MBHA Resin
With Fmoc-Ala-rink (0.04mmol), HBTU (0.04mmol), HOBT (0.04mmol) and DIPEA (0.08mmol) are dissolved among the 10mL NMP, in the resin above again this solution being added, in microwave reactor, react 7min, microwave power is 25W, and temperature of reaction is controlled at 50 ℃, uses the cooling of air compressor pressurized air.Reaction finishes back filtering reaction solution, uses DCM and each 7mL washing resin of NMP 3 times.
(4) detection of coupling efficiency
With the coupling efficiency of ninhydrin method or bromjophenol blue method qualitative detection resin, color reaction is negative can to enter next coupling circulation.
Ninhydrin method: the resin particle washing with alcohol takes a morsel, put into transparent bottle and add respectively 2 of 5% triketohydrindene hydrate ethanol, KCN pyridine solution (2ml 0.001MKCN is diluted in the 98ml pyridine), 80% phenol ethanolic solns, in 100 ℃ of heating 5 minutes, promptly positive if resin shows blueness.
The bromjophenol blue method: the resin particle that takes a morsel washs with two formyl ethanamides, puts into the tetrabromophenol sulfonphthalein dimethylacetamide solution that transparent bottle adds 3 1%, and jolting is 3 minutes under the normal temperature, and is promptly positive if resin shows blueness.
(5) prolongation of peptide chain
According to Ala 3-MC-JJ0104-(1～11)-NH 2Sequence, the steps in sequence that repeats above-mentioned deprotection and coupling is connected corresponding amino acid, the coupling microwave promotes reaction times 5～20min not wait.Obtain being connected with the resin of Ala3-MC-JJ0104-(1～11)-NH2.
(6) cracking of polypeptide on the resin
With above-mentioned obtain be connected with Ala 3The resin of-MC-JJ0104-(1～11)-NH2 is put into reaction flask, and each adds cracking agent Reagent K, and (TFA/ thioanisole/water/phenol/EDT, 82.5: 5: 5: 5: 2.5, V/V) 10mL earlier at 0 ℃ of following jolting 30min, reacted 3h more at normal temperatures.Reaction finishes the back suction filtration, adds a small amount of TFA and DCM washing three times, merging filtrate.Filtrate added in a large amount of ice ether separate out white flocks, frozen centrifugation obtains the crude product of target polypeptides.Finally obtain Ala 3-MC-JJ0104-(1～11)-NH2 crude product 63.2mg, yield is 94.3%.
(7) Ala 3The purifying of-MC-JJ0104-(1～11)-NH2-NH2 crude product
With the Ala that obtains above 3-MC-JJ0104-(1～11)-NH2 crude product is dissolved in a spot of water, prepares type reversed-phase HPLC purifying crude product with Tianjin, island.Adopt the anti-phase preparative column of C18 (340mm * 28mm, 5 μ m) in the purifying; Mobile phase A: 0.1%TFA/ water (V/V), Mobile phase B: 0.1%TFA/ acetonitrile (V/V); Eluent gradient: Mobile phase B 13%～15%, 30min; Flow velocity is 6mL/min; The detection wavelength is 214nm.The solution freeze-drying of collecting gets pure product, finally obtains pure product 29.7mg.Theoretical relative molecular mass 907.0ESI-MSm/z:found[M+3H] 3+303.4, [M+4H] 4+227.7, [M+5H] 5+182.2; Calu[M+3H] 3+303.3, [M+4H] 4+227.7, [M+5H] 5+182.4
According to embodiment 1 described method, according to the synthetic polypeptide that obtains embodiment 2～38 of corresponding sequence, by electrospray ionization mass spectrum (ESI-MS) conclusive evidence molecular weight separately.
Embodiment 2
Gly-Pro-(D-Ala)-Ala-Tyr-Ile-Ser-Ala-Ala-Ser (SEQ.ID NO:3); Theoretical relative molecular mass 907.0ESI-MSm/z:found[M+3H] 3+303.5, [M+4H] 4+227.6, [M+5H] 5+182.2; Calu[M+3H] 3+303.3, [M+4H] 4+227.7, [M+5H] 5+182.4
Embodiment 3
Gly-Pro-Ala-(D-Ala)-Tyr-Ile-Ser-Ala-Ala-Ser (SEQ.ID NO:4); Theoretical relative molecular mass 907.0ESI-MSm/z:found[M+3H] 3+303.3, [M+4H] 4+227.5, [M+5H] 5+182.4; Calu[M+3H] 3+303.3, [M+4H] 4+227.7, [M+5H] 5+182.4
Embodiment 4
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-(D-Ala)-Ala-Ser (SEQ.ID NO:5); Theoretical relative molecular mass 907.0ESI-MSm/z:found[M+3H] 3+303.4, [M+4H] 4+227.7, [M+5H] 5+182.4; Calu[M+3H] 3+303.3, [M+4H] 4+227.7, [M+5H] 5+182.4
Embodiment 5
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-(D-Ala)-Ser (SEQ.ID NO:6); Theoretical relative molecular mass 907.0ESI-MSm/z:found[M+3H] 3+303.3[M+4H] 4+227.7, [M+5H] 5+182.5; Calu[M+3H] 3+303.3, [M+4H] 4+227.7, [M+5H] 5+182.4
Embodiment 6
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Gly-Ser (SEQ.ID NO:7); Theoretical relative molecular mass 893.0ESI-MSm/z:found[M+3H] 3+298.7[M+4H] 4+224.2, [M+5H] 5+179.6; Calu[M+3H] 3+298.7, [M+4H] 4+224.2, [M+5H] 5+179.6
Embodiment 7
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Cys-Ser (SEQ.ID NO:8); Theoretical relative molecular mass 939.0ESI-MSm/z:found[M+3H] 3+314.1[M+4H] 4+235.8, [M+5H] 5+188.8; Calu[M+3H] 3+314.0, [M+4H] 4+235.8, [M+5H] 5+188.8
Embodiment 8
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Arg-Ser (SEQ.ID NO:9); Theoretical relative molecular mass 992.1ESI-MSm/z:found[M+3H] 3+331.7[M+4H] 4+249.1, [M+5H] 5+199.5; Calu[M+3H] 3+331.7, [M+4H] 4+249.0, [M+5H] 5+199.4
Embodiment 9
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Met-Ser (SEQ.ID NO:10); Theoretical relative molecular mass 967.1ESI-MSm/z:found[M+3H] 3+323.5[M+4H] 4+249.1, [M+5H] 5+199.5; Calu[M+3H] 3+331.7, [M+4H] 4+249.0, [M+5H] 5+199.4
Embodiment 10
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Ser-Ser (SEQ.ID NO:11); Theoretical relative molecular mass 923.0ESI-MSm/z:found[M+3H] 3+308.7[M+4H] 4+231.8, [M+5H] 5+185.5; Calu[M+3H] 3+308.7, [M+4H] 4+231.7, [M+5H] 5+185.6
Embodiment 11
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Lys-Ser (SEQ.ID NO:12); Theoretical relative molecular mass 964.1ESI-MSm/z:found[M+3H] 3+322.4[M+4H] 4+242.1, [M+5H] 5+193.7; Calu[M+3H] 3+322.4, [M+4H] 4+242.0, [M+5H] 5+193.8
Embodiment 12
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:13); Theoretical relative molecular mass 1021.2ESI-MSm/z:found[M+3H] 3+341.4[M+4H] 4+256.1, [M+5H] 5+205.2; Calu[M+3H] 3+341.4, [M+4H] 4+256.3, [M+5H] 5+205.2
Embodiment 13
Gly-Pro-(D-Ala)-Ala-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:14); Theoretical relative molecular mass 950..1ESI-MSm/z:found[M+3H] 3+317.5[M+4H] 4+238.5, [M+5H] 5+191.2; Calu[M+3H] 3+317.7, [M+4H] 4+238.5, [M+5H] 5+191.0
Embodiment 14
Gly-Pro-Ala-(D-Ala)-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:15); Theoretical relative molecular mass 1021.2ESI-MSm/z:found[M+3H] 3+341.5[M+4H] 4+256.5, [M+5H] 5+205.2; Calu[M+3H] 3+341.4, [M+4H] 4+256.3, [M+5H] 5+205.2
Embodiment 15
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Gly-Lys-Ser (SEQ.ID NO:16); Theoretical relative molecular mass 950.0ESI-MSm/z:found[M+3H] 3+317.5[M+4H] 4+238.5, [M+5H] 5+191.0; Calu[M+3H] 3+317.7, [M+4H] 4+238.5, [M+5H] 5+191.0
Embodiment 16
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Tyr-Lys-Ser (SEQ.ID NO:17); Theoretical relative molecular mass 1021.2ESI-MSm/z:found[M+3H] 3+341.5[M+4H] 4+256.5, [M+5H] 5+205.0; Calu[M+3H] 3+341.4, [M+4H] 4+256.3, [M+5H] 5+205.2
Embodiment 17
Gly-Pro-Gly-Ala-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:18); Theoretical relative molecular mass 1007.1ESI-MSm/z:found[M+3H] 3+336.5[M+4H] 4+252.8, [M+5H] 5+202.4; Calu[M+3H] 3+336.7, [M+4H] 4+252.8, [M+5H] 5+202.4
Embodiment 18
Gly-Pro-Ser-Ala-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:19); Theoretical relative molecular mass 1037.2ESI-MSm/z:foun d[M+3H] 3+346.7[M+4H] 4+260.3, [M+5H] 5+208.3; Calu[M+3H] 3+346.7, [M+4H] 4+260.3, [M+5H] 5+208.4
Embodiment 19
Gly-Pro-Ala-Arg-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:20); Theoretical relative molecular mass 1106.3ESI-MSm/z:found[M+3H] 3+369.8[M+4H] 4+277.7, [M+5H] 5+222.3; Calu[M+3H] 3+369.8, [M+4H] 4+277.6, [M+5H] 5+222.3
Embodiment 20
Gly-Pro-Ala-Cys-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:21); Theoretical relative molecular mass 1053.2ESI-MSm/z:found[M+3H] 3+352.1[M+4H] 4+264.3, [M+5H] 5+211.7; Calu[M+3H] 3+352.1, [M+4H] 4+264.3, [M+5H] 5+211.6
Embodiment 21
Gly-Pro-Ala-Gly-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:22); Theoretical relative molecular mass 1007.1ESI-MSm/z:found[M+3H] 3+336.6[M+4H] 4+252.9, [M+5H] 5+202.4; Calu[M+3H] 3+336.7, [M+4H] 4+252.8, [M+5H] 5+202.4
Embodiment 22
Gly-Pro-Ala-His-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:23); Theoretical relative molecular mass 1087.2ESI-MSm/z:found[M+3H] 3+363.4[M+4H] 4+272.9, [M+5H] 5+218.4; Calu[M+3H] 3+363.4, [M+4H] 4+272.8, [M+5H] 5+218.4
Embodiment 23
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:24); Theoretical relative molecular mass 1113.3ESI-MSm/z:found[M+3H] 3+372.2[M+4H] 4+279.4, [M+5H] 5+223.5; Calu[M+3H] 3+372.1, [M+4H] 4+279.3, [M+5H] 5+223.7
Embodiment 24
Gly-Pro-Ala-Ser-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:25); Theoretical relative molecular mass 1037.2ESI-MSm/z:foun d[M+3H] 3+346.7[M+4H] 4+260.4, [M+5H] 5+208.5; Calu[M+3H] 3+346.7, [M+4H] 4+260.3, [M+5H] 5+208.4
Embodiment 25
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Gly-Ser (SEQ.ID NO:26); Theoretical relative molecular mass 985.0ESI-MSm/z:found[M+3H] 3+329.2[M+4H] 4+247.4, [M+5H] 5+198.0; Calu[M+3H] 3+329.3, [M+4H] 4+247.3, [M+5H] 5+198.0
Embodiment 26
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Cys-Ser (SEQ.ID NO:27); Theoretical relative molecular mass 1031.1ESI-MSm/z:found[M+3H] 3+344.7[M+4H] 4+258.9, [M+5H] 5+207.1; Calu[M+3H] 3+344.7, [M+4H] 4+258.8, [M+5H] 5+207.2
Embodiment 27
Gly-Pro-Ala-Ty-Tyr-Ile-Ser-Ala-Met-Ser (SEQ.ID NO:28); Theoretical relative molecular mass 1059.2ESI-MSm/z:found[M+3H] 3+354.2[M+4H] 4+265.9, [M+5H] 5+212.7; Calu[M+3H] 3+354.1, [M+4H] 4+265.8, [M+5H] 5+212.8
Embodiment 28
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Ser-Ser (SEQ.ID NO:29); Theoretical relative molecular mass 1015.1ESI-MSm/z:found[M+3H] 3+339.2[M+4H] 4+254.9, [M+5H] 5+204.1; Calu[M+3H] 3+339.4, [M+4H] 4+254.8, [M+5H] 5+204.0
Embodiment 29
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Lys-Ser (SEQ.ID NO:30); Theoretical relative molecular mass 1056.2ESI-MSm/z:found[M+3H] 3+353.2[M+4H] 4+265.0, [M+5H] 5+212.1; Calu[M+3H] 3+353.1, [M+4H] 4+265.0, [M+5H] 5+212.2
Embodiment 30
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:31); Theoretical relative molecular mass 1113.3ESI-MSm/z:found[M+3H] 3+372.2[M+4H] 4+279.4, [M+5H] 5+223.6; Calu[M+3H] 3+372.1, [M+4H] 4+279.3, [M+5H] 5+223.7
Embodiment 31
Gly-Pro-Gly-(D-Ala)-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:32); Theoretical relative molecular mass 1007.1ESI-MSm/z:found[M+3H] 3+336.7[M+4H] 4+252.9, [M+5H] 5+202.6; Calu[M+3H] 3+336.7, [M+4H] 4+252.8, [M+5H] 5+202.4
Embodiment 32
Gly-Pro-Ser-(D-Ala)-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:33); Theoretical relative molecular mass 1037.2ESI-MSm/z:found[M+3H] 3+346.7[M+4H] 4+260.1, [M+5H] 5+208.6; Calu[M+3H] 3+346.7, [M+4H] 4+260.3, [M+5H] 5+208.4
Embodiment 33
Gly-Pro-(D-Ala)-Arg-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:34); Theoretical relative molecular mass 1106.3ESI-MSm/z:found[M+3H] 3+369.7[M+4H] 4+277,5[M+5H] 5+222.2; Calu[M+3H] 3+369.8, [M+4H] 4+277.6, [M+5H] 5+208.4
Embodiment 34
Gly-Pro-(D-Ala)-Cys-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:35); Theoretical relative molecular mass 1053.2ESI-MSm/z:found[M+3H] 3+352.2[M+4H] 4+264,5[M+5H] 5+211.5; Calu[M+3H] 3+352.1, [M+4H] 4+264.3, [M+5H] 5+211.6
Embodiment 35
Gly-Pro-(D-Ala)-Gly-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:36); Theoretical relative molecular mass 1007.1ESI-MSm/z:found[M+3H] 3+336.6[M+4H] 4+252.8[M+5H] 5+202.5; Calu[M+3H] 3+336.7, [M+4H] 4+252.8, [M+5H] 5+202.4
Embodiment 36
Gly-Pro-(D-Ala)-His-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:37); Theoretical relative molecular mass 1087.2ESI-MSm/z:found[M+3H] 3+363.4[M+4H] 4+272.8[M+5H] 5+218.5; Calu[M+3H] 3+363.4, [M+4H] 4+272.8, [M+5H] 5+218.4
Embodiment 37
Gly-Pro-(D-Ala)-Tyr-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:38); Theoretical relative molecular mass 1087.2ESI-MSm/z:found[M+3H] 3+363.5[M+4H] 4+272.7[M+5H] 5+218.5; Calu[M+3H] 3+363.4, [M+4H] 4+272.8, [M+5H] 5+218.4
Embodiment 38
Gly-Pro-(D-Ala)-Ser-Tyr-Ile-Ser-Lys-Lys-Ser (SEQ.ID NO:39); Theoretical relative molecular mass 1037.2ESI-MSm/z:found[M+3H] 3+346.5[M+4H] 4+260.4[M+5H] 5+208.5; Calu[M+3H] 3+346.7, [M+4H] 4+260.3, [M+5H] 5+208.4
Embodiment 39
Hypoglycemic activity experiment in MC-JJ0104 and the similar object thereof
Get 10 the week age male mouse of kunming (body weight 18～22g), random packet, 6 every group.Tail vein injection tetraoxypyrimidine administration modeling, dosage is 60mg/kg, after 72 hours, mouse fasting 5～6 hours is surveyed blood sugar and is selected the blood glucose value scope 13～23 mouse random packet, 6 every group.Mouse 1mg/kg successive administration 10d abdominal injection MC-JJ0104 polypeptide and analogue thereof behind the Cheng Mo.0,30,60,90,120 usefulness blood glucose meter are measured blood glucose value.
As shown in table 1, the hypoglycemic activity of MC-JJ0104 polypeptide and analogue thereof is better than N1,N1-Dimethylbiguanide.
Table 1MC-JJ0104 and the hypoglycemic effect of analogue thereof
??0min ??30min ??60min ??90min ??120min
??SEQ.ID?NO：2 ??15.36±1.19* ??18.37±0.79* ??18.01±3.69** ??15.98±2.41** ??15.49±0.52**
??SEQ.ID?NO：4 ??18.23±3.21 ??19.35±1.58* ??18.33±2.54** ??17.55±2.48** ??16.17±3.26
??SEQ.ID?NO：7 ??17.34±2.42* ??17.22±1.37* ??16.25±1.85*** ??15.27±7.83** ??14.54±3.77**
??SEQ.ID?NO：8 ??17.54±2.69* ??19.62±9.72* ??18.37±12.15** ??15.81±12.52 ??12.27±6.91***
??SEQ.ID?NO：12 ??19.45±10.17 ??21.12±15.40* ??18.71±18.57** ??18.84±15.64* ??17.57±15.21*
??SEQ.ID?NO：15 ??16.61±2.27* ??17.40±3.94* ??16.96±6.34*** ??14.15±6.51*** ??13.30±6.53***
??SEQ.ID?NO：19 ??20.15±10.84 ??18.86±11.54 ??17.24±11.34*** ??15.95±13.94** ??11.71±11.05***
??SEQ.ID?NO：24 ??17.89±8.74* ??18.45±12.46* ??19.19±21.68* ??18.45±22.44** ??17.82±8.31*
Positive blank ??22.1±5.54 ??22.76±4.41 ??22.53±3.39 ??21.83±1.33 ??20.35±0.45
N1,N1-Dimethylbiguanide ??22.75±8.57 ??20.45±7.00 ??18.88±2.20 ??17.71±5.31 ??15.77±5.25
Negative blank ??7.43±0.58 ??8.50±0.62 ??7.38±0.90 ??7.18±2.26 ??6.51±1.49
n＝6，
Figure GSA00000127628200091
*P＜0.05， **P＜0.01， ***P＜0.001vs?saline
Sequence table
＜110〉China Medicine University
＜120〉microwave promotes solid phase synthesis balsam pear MC-JJ0104 polypeptide analog and application thereof
<160>10
 
<210>1
<211>116
<212>PRT
＜213〉artificial sequence
 
<220>
＜221〉synthetic construct
<222>(3)..(3)
＜223〉the 3rd Xaa is Ala, D-Ala, Gly or Ser;
 
<220>
＜221〉synthetic construct
<222>(4)..(4)
＜223〉the 4th Xaa is Ala, D-Ala, Arg, Cys, Gly, His, Lys, Met, Tyr or Ser;
 
<220>
＜221〉synthetic construct
<222>(8)..(8)
＜223〉the 8th Xaa is Ala, D-Ala, Gly, Lys, Tyr or Ser;
 
<220>
＜221〉synthetic construct
<222>(9)..(9)
＜223〉the 9th Xaa is Ala, D-Ala, Gly, Cys, Arg, Lys, Met or Ser;
<400>1
 
<210>2
<211>10
<212>PRT
＜213〉artificial sequence
<400>1
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Ala-Ser
1????????????????5??????????????????10
<210>3
<211>10
<212>PRT
＜213〉artificial sequence
<400>3
Gly-Pro-(D-Ala)-Ala-Tyr-Ile-Ser-Ala-Ala-Ser
1????????????????????5??????????????????10
 
<210>4
<211>10
<212>PRT
＜213〉artificial sequence
<400>4
Gly-Pro-Ala-(D-Ala)-Tyr-Ile-Ser-Ala-Ala-Ser
1????????????????????5???????????????????10
 
<210>5
<211>10
<212>PRT
＜213〉artificial sequence
<400>5
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-(D-Ala)-Ala-Ser
1????????????????5???????????????????????10
 
<210>6
<211>10
<212>PRT
＜213〉artificial sequence
<400>6
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-(D-Ala)-Ser
1????????????????5???????????????????????10
 
<210>7
<211>10
<212>PRT
＜213〉artificial sequence
<400>7
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Gly-Ser
1????????????????5??????????????????10
 
<210>8
<211>10
<212>PRT
＜213〉artificial sequence
<400>8
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Cys-Ser
1????????????????5??????????????????10
 
<210>9
<211>10
<212>PRT
＜213〉artificial sequence
<400>9
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Arg-Ser
1????????????????5??????????????????10
 
<210>10
<211>10
<212>PRT
＜213〉artificial sequence
<400>10
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Met-Ser
1????????????????5???????????????????10
 
<210>11
<211>10
<212>PRT
＜213〉artificial sequence
<400>11
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Ser-Ser
1????????????????5???????????????????10
 
<210>12
<211>10
<212>PRT
＜213〉artificial sequence
<400>12
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Lys-Ser
1????????????????5???????????????????10
 
<210>13
<211>10
<212>PRT
＜213〉artificial sequence
<400>13
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????5???????????????????10
<210>14
<211>10
<212>PRT
＜213〉artificial sequence
<400>14
Gly-Pro-(D-Ala)-Ala-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????????5???????????????????10
 
<210>15
<211>10
<212>PRT
＜213〉artificial sequence
<400>15
Gly-Pro-Ala-(D-Ala)-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????????5???????????????????10
 
<210>16
<211>10
<212>PRT
＜213〉artificial sequence
<400>16
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Gly-Lys-Ser
1????????????????5????????????????????10
 
<210>17
<211>10
<212>PRT
＜213〉artificial sequence
<400>17
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Tyr-Lys-Ser
1????????????????5???????????????????10
 
<210>18
<211>10
<212>PRT
＜213〉artificial sequence
<400>18
Gly-Pro-Gly-Ala-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????5???????????????????10
 
<210>19
<211>10
<212>PRT
＜213〉artificial sequence
<400>19
Gly-Pro-Ser-Ala-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????5????????????????????10
 
<210>20
<211>10
<212>PRT
＜213〉artificial sequence
<400>20
Gly-Pro-Ala-Arg-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????5??????????????????10
 
<210>21
<211>10
<212>PRT
＜213〉artificial sequence
<400>21
Gly-Pro-Ala-Cys-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????5???????????????????10
 
<210>22
<211>10
<212>PRT
＜213〉artificial sequence
<400>22
Gly-Pro-Ala-Gly-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????5???????????????????10
 
<210>23
<211>10
<212>PRT
＜213〉artificial sequence
<400>23
Gly-Pro-Ala-His-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????5???????????????????10
 
<210>24
<211>10
<212>PRT
＜213〉artificial sequence
<400>24
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????5??????????????????10
<210>25
<211>10
<212>PRT
＜213〉artificial sequence
<400>25
Gly-Pro-Ala-Ser-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????5??????????????????10
 
<210>26
<211>10
<212>PRT
＜213〉artificial sequence
<400>26
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Gly-Ser
1????????????????5???????????????????10
 
<210>27
<211>10
<212>PRT
＜213〉artificial sequence
<400>27
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Cys-Ser
1????????????????5???????????????????10
 
<210>28
<211>10
<212>PRT
＜213〉artificial sequence
<400>28
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Met-Ser
1????????????????5???????????????????10
 
<210>29
<211>10
<212>PRT
＜213〉artificial sequence
<400>29
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Ser-Ser
1????????????????5???????????????????10
 
<210>30
<211>10
<212>PRT
＜213〉artificial sequence
<400>30
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Lys-Ser
1????????????????5???????????????????10
 
<210>31
<211>10
<212>PRT
＜213〉artificial sequence
<400>31
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????5???????????????????10
 
<210>32
<211>10
<212>PRT
＜213〉artificial sequence
<400>32
Gly-Pro-Gly-(D-Ala)-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????????5???????????????????10
 
<210>33
<211>10
<212>PRT
＜213〉artificial sequence
<400>33
Gly-Pro-Ser-(D-Ala)-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????????5???????????????????10
 
<210>34
<211>10
<212>PRT
＜213〉artificial sequence
<400>34
Gly-Pro-(D-Ala)-Arg-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????????5???????????????????10
 
<210>35
<211>10
<212>PRT
＜213〉artificial sequence
<400>35
Gly-Pro-(D-Ala)-Cys-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????????5???????????????????10
<210>36
<211>10
<212>PRT
＜213〉artificial sequence
<400>36
Gly-Pro-(D-Ala)-Gly-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????????5???????????????????10
 
<210>37
<211>10
<212>PRT
＜213〉artificial sequence
<400>37
Gly-Pro-(D-Ala)-His-Tyr-Ile-Ser-Lys-Lys-Ser
1?????????????????????5???????????????????10
 
<210>38
<211>10
<212>PRT
＜213〉artificial sequence
<400>38
Gly-Pro-(D-Ala)-Tyr-Tyr-Ile-Ser-Lys-Lys-Ser
1????????????????????5???????????????????10
 
<210>39
<211>10
<212>PRT
＜213〉artificial sequence
<400>39
Gly-Pro-(D-Ala)-Ser-Tyr-Ile-Ser-Lys-Lys-Ser
1?????????????????????5???????????????????10

Claims (8)

1. balsam pear MC-JJ0104 polypeptide analog that contains formula I (SEQ.ID NO:1) structure with hypoglycemic activity, the balsam pear MC-JJ0104 polypeptide analog that obtains; It is characterized in that its structure has following form:
Gly-Pro-Xaa 1-Xaa 2-Tyr-Ile-Ser-Xaa 3-Xaa 4-Ser
(SEQ.ID?NO：1)
Xaa 1: Ala, D-Ala, Gly or Ser
Xaa 2: Ala, D-Ala, Arg, Cys, Gly, His, Lys, Met, Tyr or Ser
Xaa 3: Ala, D-Ala, Gly, Lys, Tyr or Ser
Xaa 4: Ala, D-Ala, Gly, Cys, Arg, Lys, Met or Ser
2. polypeptide according to claim 1 has following sequence:
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Ala-Ser??????(SEQ.ID?NO：2)
Gly-Pro-(D-Ala)-Ala-Tyr-Ile-Ser-Ala-Ala-Ser??(SEQ.ID?NO：3)
Gly-Pro-Ala-(D-Ala)-Tyr-Ile-Ser-Ala-Ala-Ser??(SEQ.ID?NO：4)
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-(D-Ala)-Ala-Ser??(SEQ.ID?NO：5)
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-(D-Ala)-Ser??(SEQ.ID?NO：6)
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Gly-Ser??????(SEQ.ID?NO：7)
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Cys-Ser??????(SEQ.ID?NO：8)
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Arg-Ser??????(SEQ.ID?NO：9)
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Met-Ser??????(SEQ.ID?NO：10)
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Ser-Ser??????(SEQ.ID?NO：11)
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Ala-Lys-Ser??????(SEQ.ID?NO：12)
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Lys-Lys-Ser??????(SEQ.ID?NO：13)
Gly-Pro-(D-Ala)-Ala-Tyr-Ile-Ser-Lys-Lys-Ser??(SEQ.ID?NO：14)
Gly-Pro-Ala-(D-Ala)-Tyr-Ile-Ser-Lys-Lys-Ser??(SEQ.ID?NO：15)
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Gly-Lys-Ser??????(SEQ.ID?NO：16)
Gly-Pro-Ala-Ala-Tyr-Ile-Ser-Tyr-Lys-Ser??????(SEQ.ID?NO：17)
Gly-Pro-Gly-Ala-Tyr-Ile-Ser-Lys-Lys-Ser??????(SEQ.ID?NO：18)
Gly-Pro-Ser-Ala-Tyr-Ile-Ser-Lys-Lys-Ser??????(SEQ.ID?NO：19)
Gly-Pro-Ala-Arg-Tyr-Ile-Ser-Lys-Lys-Ser??????(SEQ.ID?NO：20)
Gly-Pro-Ala-Cys-Tyr-Ile-Ser-Lys-Lys-Ser??????(SEQ.ID?NO：21)
Gly-Pro-Ala-Gly-Tyr-Ile-Ser-Lys-Lys-Ser??????(SEQ.ID?NO：22)
Gly-Pro-Ala-His-Tyr-Ile-Ser-Lys-Lys-Ser????????(SEQ.ID?NO：23)
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Lys-Lys-Ser????????(SEQ.ID?NO：24)
Gly-Pro-Ala-Ser-Tyr-Ile-Ser-Lys-Lys-Ser????????(SEQ.ID?NO：25)
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Gly-Ser????????(SEQ.ID?NO：26)
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Cys-Ser????????(SEQ.ID?NO：27)
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Met-Ser????????(SEQ.ID?NO：28)
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Ser-Ser????????(SEQ.ID?NO：29)
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Ala-Lys-Ser????????(SEQ.ID?NO：30)
Gly-Pro-Ala-Tyr-Tyr-Ile-Ser-Lys-Lys-Ser????????(SEQ.ID?NO：31)
Gly-Pro-Gly-(D-Ala)-Tyr-Ile-Ser-Lys-Lys-Ser????(SEQ.ID?NO：32)
Gly-Pro-Ser-(D-Ala)-Tyr-Ile-Ser-Lys-Lys-Ser????(SEQ.ID?NO：33)
Gly-Pro-(D-Ala)-Arg-Tyr-Ile-Ser-Lys-Lys-Ser????(SEQ.ID?NO：34)
Gly-Pro-(D-Ala)-Cys-Tyr-Ile-Ser-Lys-Lys-Ser????(SEQ.ID?NO：35)
Gly-Pro-(D-Ala)-Gly-Tyr-Ile-Ser-Lys-Lys-Ser????(SEQ.ID?NO：36)
Gly-Pro-(D-Ala)-His-Tyr-Ile-Ser-Lys-Lys-Ser????(SEQ.ID?NO：37)
Gly-Pro-(D-Ala)-Tyr-Tyr-Ile-Ser-Lys-Lys-Ser????(SEQ.ID?NO：38)
Gly-Pro-(D-Ala)-Ser-Tyr-Ile-Ser-Lys-Lys-Ser????(SEQ.ID?NO：39)
3. a pharmaceutical composition comprises the MC-JJ0104 polypeptide analog described in the claim 1 for the treatment of significant quantity or its pharmacy acceptable salt and pharmaceutically acceptable carrier or thinner.
4. the MC-JJ0104 polypeptide analog described in the claim 1 or its pharmacy acceptable salt and pharmaceutically acceptable carrier or thinner are used for the treatment of utilization in the medicine of diabetes in preparation.
5. the preparation method of the MC-JJ0104 polypeptide analog described in the claim 1, it is characterized in that: adopt microwave to promote Fmoc/tBu orthogonally protect solid phase synthesis strategy, earlier syntheticly on solid phase carrier obtain being loaded with first Fmoc and protect amino acid whose resin, ninhydrin method detects sloughs the resin that the Fmoc protecting group obtains being loaded with first amino-acid residue after negative; Enter next coupling circulation again; repeat the step of coupling and deprotection with different protection amino acid according to corresponding peptide preface; prolong required aminoacid sequence successively, the synthetic resin that obtains being loaded with corresponding polypeptide cuts down polypeptide with cracking agent at last and obtains the polypeptide crude product from resin.Crude product is through the preparation scale high-efficient liquid phase chromatogram purification, and freeze-drying gets the pure product of polypeptide.
6. promote solid phase synthesis MC-JJ0104 polypeptide analog method according to microwave described in the claim 5; it is characterized in that being loaded with the preparation method that first Fmoc protects amino acid whose resin; be by obtaining with the solid phase carrier coupling again after the activated dose of activation earlier of Fmoc protection amino acid under the microwave irradiation condition; add 1-hydroxyl-benzotriazole (HOBT) or derivatives thereof in the reaction and suppress racemization, and use the acidity of organic bases neutralization reaction.
7. according to claim 6ly be loaded with the preparation method that first Fmoc protects amino acid whose resin, it is characterized in that the solid phase carrier that uses is Rink resin, Wang resin or 2-chlorine trityl chloride resin; The activator that uses is dicyclohexylcarbodiimide (DCC), N, N '-DIC (DIC), N, N " carbonyl dimidazoles (CDI), 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC.HCl), 2-(7-azo benzotriazole)-N; N; N '; N '-tetramethyl-urea phosphofluoric acid ester (HATU), benzotriazole-N; N; N ', N '-tetramethyl-urea phosphofluoric acid ester (HBTU) or 6-chlorobenzene and triazole-1,1,3,3-tetramethyl-urea phosphofluoric acid ester (HCTU); 1-hydroxyl-benzotriazole (HOBT) derivative that uses is N-hydroxy-succinamide (HOSU), 1-hydroxyl-7-azo benzotriazole (HOAT) or 3-hydroxyl-1,2,3-phentriazine-4 (3H)-ketone (HOOBT); The organic bases that uses is triethylamine (TEA), N-methylmorpholine (NMM) or diisopropylethylamine (DIEA); Microwave promotion condition is: magnetron frequency 2450MHz, temperature of reaction: 20～100 ℃, the reaction times is 5～15min.
8. promote solid phase synthesis MC-JJ0104 polypeptide analog synthetic method according to microwave described in the claim 5, it is characterized in that removing of Fmoc protecting group is to contain 0.1mol.L by use -1Hexahydropyridine solution reaction under microwave promotes of 1-hydroxyl-benzotriazole (HOBT), selecting dimethyl formamide (DM F), methyl-sulphoxide (DM SO) or N-Methyl pyrrolidone (NMP) for use is reaction solvent; Microwave promotion condition is: magnetron frequency 2450MHz, temperature of reaction: 20～100 ℃, the reaction times is 1～10min.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102286078A (en) * 2011-07-13 2011-12-21 中国药科大学 Method for preparing polypeptide HM-3

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1255044A (en) * 1997-04-01 2000-05-31 卡里克斯治疗公司 Orally active fraction of momordica charactia, active peptides thereof, and their use in the treatment of diabetes

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1255044A (en) * 1997-04-01 2000-05-31 卡里克斯治疗公司 Orally active fraction of momordica charactia, active peptides thereof, and their use in the treatment of diabetes

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102286078A (en) * 2011-07-13 2011-12-21 中国药科大学 Method for preparing polypeptide HM-3

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