CN102174084B - Thymosin α1 active fragments cyclic peptide analogue and polyethylene glycol derivative thereof - Google Patents

Thymosin α1 active fragments cyclic peptide analogue and polyethylene glycol derivative thereof Download PDF

Info

Publication number
CN102174084B
CN102174084B CN201110034234.4A CN201110034234A CN102174084B CN 102174084 B CN102174084 B CN 102174084B CN 201110034234 A CN201110034234 A CN 201110034234A CN 102174084 B CN102174084 B CN 102174084B
Authority
CN
China
Prior art keywords
ring
peg
cys
compound
peptide
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CN201110034234.4A
Other languages
Chinese (zh)
Other versions
CN102174084A (en
Inventor
刘克良
蒋志龙
王良友
梁远军
许笑宇
吴萍
韩寒
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Institute of Pharmacology and Toxicology of AMMS
Original Assignee
Institute of Pharmacology and Toxicology of AMMS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Institute of Pharmacology and Toxicology of AMMS filed Critical Institute of Pharmacology and Toxicology of AMMS
Publication of CN102174084A publication Critical patent/CN102174084A/en
Application granted granted Critical
Publication of CN102174084B publication Critical patent/CN102174084B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Abstract

The present invention relates to the cyclic peptide derivative that a class contains the thymosin α1 active fragments that natural or alpha-non-natural amino acid replaces, its preparation method, containing their pharmaceutical composition, and they are at the medicine of the relative diseases such as treatment or epidemic prevention defect, immunologic hypofunction.

Description

Thymosin α1 active fragments cyclic peptide analogue and polyethylene glycol derivative thereof
The divisional application of the present invention's to be application number be female case of 200410096459.2, the applying date of this female case is on December 1st, 2004, and denomination of invention is " thymosin α1 active fragments cyclic peptide analogue and polyethylene glycol derivative thereof ".
Invention field
The present invention relates to thymosin α1 active fragments and cyclic peptide analogue that natural or alpha-non-natural amino acid replaces, and polyethylene glycol derivative, and preparation method thereof, containing their pharmaceutical composition and the purposes in the middle for the treatment of or prevention and the diseases related such as immune deficiency, immunologic hypofunction thereof.
Background technology:
Thymosin α1 (T α 1) is a kind of important polypeptide para-immunity regulatory factor, it is a kind of for the lymphocytic immunostimulant of T for having the significant effect strengthening immunologic function, maturation and the differentiation of T cell can be promoted, and impel the ripe multiple lymphokine of lymphocytic emiocytosis (as interleukin-2 and gamma-interferon etc.) that the generation of Interleukin 2 Receptor can also be promoted.T α 1 is made up of 28 amino-acid residues, molecular weight 3108, and iso-electric point 4.2, N-is held as acetylize structure, and its aminoacid sequence is as follows:
Ac-Ser-Asp-Ala-Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-
Thr-Thr-Lys-Asp-Leu-Lys-Glu-Lys-Lys-Glu-Val-Val
-Glu-Glu-Ala-Glu-Asp-OH
In recent years, the research of T α 1 mainly concentrates on clinical application, and the research report about its structure activity relationship is few.1980, Wong etc. synthesized T α 1with desAc-T α 1, suppress their activity of E-RFT display and natural T α by external azathioprine 1quite, T α is described 1n-hold acetylize whether active in impact on it.Except report T α such as Frasca 1n-end fragment 1-14 can promote IL-2 expression of receptor in adult rat (the 6-18 month) body, strengthen helper T cell activity outside, other research shows T α 1the essentially no activity of N-end fragment, C-end portion fragment has good activity.The external E-RFT such as Abiko confirms T α 1some C-end fragment have amount of activated: T α 1 (100%), 14-28 (9.83%), 19-28 (1.04%).The immunosuppressed mice test in body such as Felix confirms the basic non-activity of N-end fragment 1-4,4-10,1-10 and 1-14; C-end fragment 15-28 and 18-28 and T α 1quite active; 21-28 (75%), 23-28 (50%) and 25-28 (50%) have amount of activated.The external mixed lymphocytes proliferation test (MLC) such as Ciaredelli confirms T α 1c-to hold 20-26,25-27 and 25-28 to have about 90% active.Ho etc. report C-end fragment 13-19 and T α 1have the activity of identical short Cord blood T lymphocyte differentiation maturation in vitro, 20-25 and 20-28 has amount of activated.Ciaredelli etc. suppress E-RFT to find with external sulphur azoles purine, and C-holds 17-28 area part fragment to have good activity.
Grottesi etc. by circular dichroism spectrum (CD) and 2D-NMR to T α 1conformation is in the solution studied, and result shows: T α 1do not form the secondary con of advantage in aqueous, but can certain secondary structure be formed in hydrophobic environment.In trifluoroethanol (TFE) aqueous solution 40%, Val 5and Glu 8between form β-corner, Lys 17and Glu 24between form alpha-helix.Due to T α 1receptor structure is also not yet found, and Grottesi etc. infer T α 1with lymphocytic mechanism of action may be: T α 1first form electrostatic interaction by polar amino acid residues and cytolemma, then induce Lys 17to Glu 24between sequence form α-helixstructure embed cytolemma, and then there is electrostatic interaction with IL-2 receptor transmembrane district.This action model contributes to explaining T α 1with the synergy of IL-2 in biological activity.
Type B viral hepatitis and hepatitis C can cause liver cirrhosis to cause liver cancer, and the whole world has 3.5 hundred million hepatitis b virus carriers and 1.5 hundred million hepatitis C carrier at present.China is hepatitis B big country, has the hepatitis B carriers of more than 1.2 hundred million, has the hepatitis B patient of more than 30,000,000, the T α of chemosynthesis 1(Zadaxin) in 1996 first in Discussion on Chinese Listed, be used alone or with alpha-interferon conbined usage Treatment chronic Hepatitis B.The curative effect of Zadaxin in treatment hepatitis B is equal to or is better than existing ethical goods.Zadaxin has certain curative effect for many Other diseases (as hepatitis C, nonsmall-cell lung cancer, melanoma and acquired immune deficiency syndrome (AIDS) etc.).In addition, Zadaxin also can be used as vaccine adjuvant, strengthens the immune effect of influenza vaccines and hepatitis B vaccine.At present, Zadaxin just carries out treatment 3 clinical trial phases of HCV and 2 clinical trial phases of Hepatoma therapy in the U.S., and 3 clinical trial phases for the treatment of HBV Japan carry out, and 2 clinical trial phases for the treatment of malignant melanoma carry out in Europe.As can be seen here, research T α 1 and analogue thereof, find more efficient immunostimulant and antiviral drug has good prospect.
The T α 1 (Zadaxin) of current Clinical practice is synthetic, expensive, dosage is large, the cycle is long, therefore carries out structure activity study and finds that better analogue has good application prospect.The same with other peptide medicament, T α 1 have the such as transformation period short, the shortcoming such as to be easily degraded in vivo, therefore the T α 1 analogue tool of active high, the anti-vivo degradation of design is of great significance.
Summary of the invention:
The present inventor and place laboratory study thereof have now found that T α 1 and derivative thereof have good prevention or treat the biological activity of immune deficiency, immunologic hypofunction diseases related, the derivative that they and polyoxyethylene glycol are formed is while can keeping activity, extend following (I), (IV), the transformation period of compound that (VII), (VIII) organize metabolism in vivo, thus reduce following (I), (IV), the consumption of compound that (VII), (VIII) organize and extend its action time in vivo.
Concrete, the present invention relates to T α 1 active fragments derivative as follows and steric isomer thereof.
One aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, and it is the compound that (I) organizes:
X 1-Y 1-Y 2-X 2
X 1-X 2-Y 1-Y 2-X 3
X 1-X 2-X 3-Y 1-Y 2-X 4
X 1-X 2-Y 1-Y 2-X 3-X 4
X 1-X 2-X 3-Y 1-Y 2-X 4-X 5
X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3
X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3
X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5
X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6
X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-Y 4
X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4
X 1-X 2-X 3-X 4-Y 1-Y 2-X 5
X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6
X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7
X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7
X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y3-Y4-X 7(I)
Wherein, X 1, X 2, X 3, X 4, X 5, X 6, X 7be the natural of L-or D-type or non-natural alkalescence, acidity or aromatic amino acid and derivative thereof independently, described basic aminoacids is selected from Arg, Lys, Glu, Cys, His, Trp, Mob, Nal, Pya, Phe (X) and Phg (X), wherein X is optional monosubstituted or two replacements of hydrogen, halogen, nitro, carboxyl or C1-C4 alkyl, substituting group be selected from H, 4-F, 3-F, 2-F, 4-Cl, 2-Cl, 4-Br, 2-Br, 3-Br, 2,5-2Cl, 4-F, 3-Cl, 3-NO 2; Acidic amino acid is selected from: Asp, Glu, His, Tyr, Trp, Mob, Nal, Pya and Phg (X), wherein X is optional monosubstituted or two replacements of hydrogen, halogen, nitro, carboxyl or C1-C4 alkyl, substituting group be selected from H, 4-F, 3-F, 2-F, 4-Cl, 2-Cl, 4-Br, 2-Br, 3-Br, 2,5-2Cl, 4-F, 3-Cl, 3-NO 2;
Y 1, Y 2, Y 3, Y 4be independently the natural of L-or D-type or non-natural is neutral, lipotropy or aromatic amino acid and derivative thereof, be selected from Gly, Ala, β-Ala, GABA, Val, Leu, Ile, Pro, His, Tyr, Trp, Mob, Nal, Pya, Phe (X), Phg (X), wherein X is optional monosubstituted or two replacements of hydrogen, halogen, nitro, carboxyl or C1-C4 alkyl, substituting group be selected from H, 4-F, 3-F, 2-F, 4-Cl, 2-Cl, 4-Br, 2-Br, 3-Br, 2,5-2Cl, 4-F, 3-Cl, 3-NO 2.
Another aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, in compound wherein in above-mentioned (I) group, the amino in any site is modified through PEG, become the following compound shown in (II) group, wherein said amino comprises N-Amino End Group and amino acid side chain is amino:
[PEG-X-(CH 2) MCO-NH] Z-X 1-Y 1-Y 2-X 2
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-Y 1-Y 2-X 3
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-X 3-Y 1-Y 2-X 4
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-Y 1-Y 2-X 3-X 4
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-Y 4
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7
[PEG-X-(CH 2) MCO-NH] Z-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y3-Y4-X 7(II)
Wherein PEG represents: RO (CH 2cH 2o) n-CH 2cH 2, R=H or C and CH 3, wherein n=5-1000; X=O, NH or NHCO; M=0-6; Z=1-3, X 1, X 2, X 3, X 4, X 5, X 6, X 7, Y 1, Y 2, Y 3, Y 4definition as noted above.
Another aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, wherein (I) organizes the carboxyl in any site of compound is the compound that (III) as follows organizes through PEG covalent modification, wherein said carboxyl comprises C-end carboxyl, aspartic acid and glutamate side chain carboxyl:
[PEG-X-CO] z-X 1-Y 1-Y 2-X 2
[PEG-X-CO] z-X 1-X 2-Y 1-Y 2-X 3
[PEG-X-CO] z-X 1-X 2-X 3-Y 1-Y 2-X 4
[PEG-X-CO] z-X 1-X 2-Y 1-Y 2-X 3-X 4
[PEG-X-CO] z-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5
[PEG-X-CO] z-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3
[PEG-X-CO] z-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3
[PEG-X-CO] z-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5
[PEG-X-CO] z-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6
[PEG-X-CO] z-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-Y 4
[PEG-X-CO] z-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4
[PEG-X-CO] z-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5
[PEG-X-CO] z-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6
[PEG-X-CO] z-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7
[PEG-X-CO] z-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7
[PEG-X-CO] z-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y3-Y4-X 7(III)
Wherein X 1, X 2, Y 1, Y 2, X 3, X 4, X 5, X 6, X 7, Y 1, Y 2, Y 3, Y 4definition as shown in aforementioned, X=O or NH, Z=1-3.
Another aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, wherein organize optional position in peptide sequence shown in compound at (I), comprise N end and C hold and peptide fragment in introduce halfcystine, form the compound that (IV) as follows organizes:
Cys-X 1-Y 1-Y 2-X 2
Cys-X 1-X 2-Y 1-Y 2-X 3
Cys-X 1-X 2-X 3-Y 1-Y 2-X 4
Cys-X 1-X 2-Y 1-Y 2-X 3-X 4
Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5
Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3
Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3
Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5
Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6
Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-Y 4
Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4
Cys-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5
Cys-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6
Cys-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7
Cys-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7
Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7(IV)
Another aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, the method of its compound described (IV) organized further through being selected from PEG-MAL, PEG-VS and PEG-IODS forms the compound that (V) as follows organizes after modifying, wherein PEG can be chain, starlike, ring-type, multi-arm type structure:
PEG-M-Cys-X 1-Y 1-Y 2-X 2
PEG-M-Cys-X 1-X 2-Y 1-Y 2-X 3
PEG-M-Cys-X 1-X 2-X 3-Y 1-Y 2-X 4
PEG-M-Cys-X 1-X 2-Y 1-Y 2-X 3-X 4
PEG-M-Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5
PEG-M-Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3
PEG-M-Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3
PEG-M-Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5
PEG-M-Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6
PEG-M-Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-Y 4
PEG-M-Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4
PEG-M-Cys-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5
PEG-M-Cys-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6
PEG-M-Cys-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7
PEG-M-Cys-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7
PEG-M-Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7(V)，
Wherein
Cys is halfcystine, is connected with M group covalency by side chain sulfur atom; X 1, X 2, Y 1, Y 2, X 3, X 4, X 5, X 6, X 7, Y 1, Y 2, Y 3, Y 4definition as aforementioned.
Another aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, the C end of the compound wherein organized at described (I) introduces electrophilic groups or nucleophilic group, and correspondence introduces contrary nucleophilic group or electrophilic groups at N end, obtain the compound that (VI) as follows organizes, wherein said nucleophilic reagent is selected from iodine negative ion, water, hydroxyl group anion, methoxyl group negative ion, ethanoyl negative ion, amino, nitrate ion, nitrite anions negative ion, azido-negative ion, cyano group negative ion, 1-proyl negative ion, dimethyl malonate negative ion, sulfydryl negative ion, methylthio group negative ion, thiocyanogen negative ion, dimethyl sulphide, front three phosphorus, described electrophilic reagent is selected from thiocarboxyl group acetyl ammonia, halogen, mineral acid, organic acid, aldehyde, active ketone, carboxylic acid, ester, nitro-compound:
Z 1-X 1-Y 1-Y 2-X 2-Z 2
Z 1-X 1-X 2-Y 1-Y 2-X 3-Z 2
Z 1-X 1-X 2-X 3-Y 1-Y 2-X 4-Z 2
Z 1-X 1-X 2-Y 1-Y 2-X 3-X 4-Z 2
Z 1-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Z 2
Z 1-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-Z 2
Z 1-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-Y 4-Z 2
Z 1-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4-Z 2
Z 1-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5-Z 2
Z 1-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6-Z 2
Z 1-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7-Z 2
Z 1-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7-Z 2
Z 1-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y3-Y4-X 7-Z 2(VI)，
Wherein Z 1and Z 2be one group of amino-acid residue or other fragment with nucleophilic group and electrophilic groups, work as Z 1with Z during nucleophilic group 2with electrophilic groups, work as Z 1with Z during electrophilic groups 2with nucleophilic group.
Preferably, above-mentioned tetrapeptide to ten one peptide derivant and steric isomer thereof, it is:
CEVVE-SCH 2CH 2CONH 2
CKEVVE-SCH 2CH 2CONH 2
CKKEVVE-SCH 2CH 2CONH 2
CKEVVEE-SCH 2CH 2CONH 2
CEKKEVVE-SCH 2CH 2CONH 2
CKEKKEVVE-SCH 2CH 2CONH 2
CKEVVEKEVVE-SCH 2CH 2CONH 2
CKEVVEEAE-SCH 2CH 2CONH 2
CKEVVEEA-SCH 2cH 2cONH 2, or
CKEVVEEAEN-SCH 2CH 2CONH 2。
Another aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, the chain-like structure unit of aliphatic chain, aliphatic ether chain, thioether chain, amino acid and other flexibility is introduced in any site of the compound wherein organized at (IV) and (VI), obtains the compound that (VII) as follows organizes:
Cys-L n-X 1-Y 1-Y 2-X 2
Cys-L n-X 1-X 2-Y 1-Y 2-X 3
Cys-L n-X 1-X 2-X 3-Y 1-Y 2-X 4
Cys-L n-X 1-X 2-Y 1-Y 2-X 3-X 4
Cys-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5
Cys-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3
Cys-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3
Cys-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5
Cys-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6
Cys-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-Y 4
Cys-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4
Cys-L n-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5
Cys-L n-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6
Cys-L n-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7
Cys-L n-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7
Cys-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y3-Y4-X 7，
Or following structure:
Z 1-L n-X 1-Y 1-Y 2-X 2-Z 2
Z 1-L n-X 1-X 2-Y 1-Y 2-X 3-Z 2
Z 1-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-Z 2
Z 1-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Z 2
Z 1-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Z 2
Z 1-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-Z 2
Z 1-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-Y 4-Z 2
Z 1-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4-Z 2
Z 1-L n-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5-Z 2
Z 1-L n-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6-Z 2
Z 1-L n-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7-Z 2
Z 1-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7-Z 2
Z 1-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y3-Y4-X 7-Z 2(VII)
Wherein, L represents the chain-like structure unit of any flexibility, n=1-4, and the site, L place shown in above general formula is not the unique site that can introduce, L can in peptide sequence optional position in addition to those at both ends.
Preferably, tetrapeptide to ten one peptide derivant of the present invention and steric isomer thereof, it comprises:
CLEVVE
CLKEVVE
CLKKEVVE
CLKEVVEE
CLEKKEVVE
CLKEKKEVVE
CLKEVVEKEVVE
CLKEVVEEAE
CLKEVVEEA
CLKEVVEEAEN
CLLEVVE
CLLKEVVE
CLLKKEVVE
CLLKEVVEE
CLLEKKEVVE
CLLKEKKEVVE
CLLKEVVEKEVVE
CLLKEVVEEAE
CLLKEVVEEA
CLLKEVVEEAEN，
Or the compound of following structure:
CLEVVE-SCH 2CH 2CONH 2
CLKEVVE-SCH 2CH 2CONH 2
CLKKEVVE-SCH 2CH 2CONH 2
CLKEVVEE-SCH 2CH 2CONH 2
CLEKKEVVE-SCH 2CH 2CONH 2
CLKEKKEVVE-SCH 2CH 2CONH 2
CLKEVVEKEVVE-SCH 2CH 2CONH 2
CLKEVVEEAE-SCH 2CH 2CONH 2
CLKEVVEEA-SCH 2CH 2CONH 2
CLKEVVEEAEN-SCH 2CH 2CONH 2
CLLEVVE-SCH 2CH 2CONH 2
CLLKEVVE-SCH 2CH 2CONH 2
CLLKKEVVE-SCH 2CH 2CONH 2
CLLKEVVEE-SCH 2CH 2CONH 2
CLLEKKEVVE-SCH 2CH 2CONH 2
CLLKEKKEVVE-SCH 2CH 2CONH 2
CLLKEVVEKEVVE-SCH 2CH 2CONH 2
CLLKEVVEEAE-SCH 2CH 2CONH 2
CLLKEVVEEA-SCH 2CH 2CONH 2
CLLKEVVEEAEN-SCH 2cH 2cONH 2, wherein, L=-NHCH 2cH 2oCH 2cOO-.
Another aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, wherein the compound that arbitrary aforementioned (I), (IV), (VII) organize is carried out cyclization with any known cyclisation method, obtains the ring compound shown in (VIII) group:
Ring (X 1-Y 1-Y 2-X 2)
Ring (X 1-X 2-Y 1-Y 2-X 3)
Ring (X 1-X 2-X 3-Y 1-Y 2-X 4)
Ring (X 1-X 2-Y 1-Y 2-X 3-X 4)
Ring (X 1-X 2-X 3-y 1-Y 2-X 4-X 5)
Ring (X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3)
Ring (X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3)
Ring (X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5)
Ring (X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6)
Ring (X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-y 4)
Ring (X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-y 4)
Ring (X 1-X 2-X 3-X 4-Y 1-Y 2-X 5)
Ring (X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6)
Ring (X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7)
Ring (X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
Ring (X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
Or following structure:
Ring (Cys-X 1-Y 1-Y 2-X 2)
Ring (Cys-X 1-X 2-Y 1-Y 2-X 3)
Ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4)
Ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4)
Ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5)
Ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3)
Ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3)
Ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5)
Ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6)
Ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-y 4)
Ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4)
Ring (Cys-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5)
Ring (Cys-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6)
Ring (Cys-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7)
Ring (Cys-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
Or following compound:
Ring (Z 1-L n-X 1-Y 1-Y 2-X 2)
Ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 3)
Ring (Z 1-L n-X 1-X 2-X 3-Y 1-Y 2-X 4)
Ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 3-X 4)
Ring (Z 1-L n-X 1-X 2-X 3-y 1-Y 2-X 4-X 5)
Ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3)
Ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-Y 4)
Ring (Z 1-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4)
Ring (Z 1-L n-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5)
Ring (Z 1-L n-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6)
Ring (Z 1-L n-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7)
Ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
Ring (Z 1-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7) (VIII)
Wherein, X 1, X 2, Y 1, Y 2, X 3, X 4, X 5, X 6, X 7, Y 1, Y 2, Y 3, Y 4definition as shown in aforementioned, Z 1definition as shown in aforementioned, L ndefinition as shown in aforementioned.
Preferably, tetrapeptide to ten one peptide derivant of the present invention and steric isomer thereof, it is following structure:
Ring (EVVE)
Ring (KEVVE)
Ring (KKEVVE)
Ring (KEVVEE)
Ring (EKKEVVE)
Ring (KEKKEVVE)
Ring (KEVVEKEVVE)
Ring (KEVVEEAE)
Ring (KEVVEEA)
Ring (KEVVEEAEN)
Or
Ring (CEVVE)
Ring (CKEVVE)
Ring (CKKEVVE)
Ring (CKEVVEE)
Ring (CEKKEVVE)
Ring (CKEKKEVVE)
Ring (CKEVVEKEVVE)
Ring (CKEVVEEAE)
Ring (CKEVVEEA)
Ring (CKEVVEEAEN)
Or:
Ring (CLEVVE)
Ring (CLKEVVE)
Ring (CLKKEVVE)
Ring (CLKEVVEE)
Ring (CLEKKEVVE)
Ring (CLKEKKEVVE)
Ring (CLKEVVEKEVVE)
Ring (CLKEVVEEAE)
Ring (CLKEVVEEA)
Ring (CLKEVVEEAEN)
Or:
Ring (CLLEVVE)
Ring (CLLKEVVE)
Ring (CLLKKEVVE)
Ring (CLLKEVVEE)
Ring (CLLEKKEVVE)
Ring (CLLKEKKEVVE)
Ring (CLLKEVVEKEVVE)
Ring (CLLKEVVEEAE)
Ring (CLLKEVVEEA)
Ring (CLLKEVVEEAEN).
Another aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, in the compound that wherein said (VIII) organizes, the amino in any site is modified through PEG, obtain the compound that (XI) as follows organizes, wherein said amino comprises the side-chain amino group of N-Amino End Group and natural amino acid or alpha-non-natural amino acid
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-Y 1-Y 2-X 2)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-Y 1-Y 2-X 3)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-X 3-Y 1-Y 2-X 4)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-Y 1-Y 2-X 3-X 4)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-X 3-y 1-Y 2-X 4-X 5)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-y 4)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-y 4)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-X 3-X 4-Y 1-Y 2-X 5)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
[PEG-X-(CH 2) mcO-NH] z-ring (X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y3-Y4-X 7)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-Y 1-Y 2-X 2)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-Y 1-Y 2-X 3)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-X 3-y 1-Y 2-X 4-X 5)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-y 4)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7)
[PEG-X-(CH 2) mcO-NH] z-ring (Cys-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
Or:
[PEG-X-(CH 2) mcO-NH] z-ring (Z 1-L n-X 1-Y 1-Y 2-X 2)
[PEG-X-(CH 2) mcO-NH] z-ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 3)
[PEG-X-(CH 2) mcO-NH] z-ring (Z 1-L n-X 1-X 2-X 3-Y 1-Y 2-X 4)
[PEG-X-(CH 2) mcO-NH] z-ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 3-X 4)
[PEG-X-(CH 2) mcO-NH] z-ring (Z 1-L n-X 1-X 2-X 3-y 1-Y 2-X 4-X 5)
[PEG-X-(CH 2) mcO-NH] z-ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3)
[PEG-X-(CH 2) mcO-NH] z-ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-Y 4)
[PEG-X-(CH 2) mcO-NH] z-ring (Z 1-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4)
[PEG-X-(CH 2) mcO-NH] z-ring (Z 1-L n-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5)
[PEG-X-(CH 2) mcO-NH] z-ring (Z 1-L n-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6)
[PEG-X-(CH 2) mcO-NH] z-ring (Z 1-L n-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7)
[PEG-X-(CH 2) mcO-NH] z-ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
[PEG-X-(CH 2) mcO-NH] z-ring (Z 1-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7) (XI),
Wherein said PEG represents: RO (CH 2cH 2o) n-CH 2cH 2, R=H or C and CH 3, n=5-1000; X=O, NH or NHCO; M=0-6; Z=1-3, X 1, X 2, Y 1, Y 2, X 3, X 4, X 5, X 6, X 7, Y 1, Y 2, Y 3, Y 4definition as shown in aforementioned, Z 1, L ndefinition as shown in aforementioned.
Another aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, comprises following compound:
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CKKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CKEVVEE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CEKKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CKEKKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CKEVVEKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CKEVVEEAE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CKEVVEEA)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CKEVVEEAEN)
Or following structure:
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLKKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLKEVVEE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLEKKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLKEKKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLKEVVEKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLKEVVEEAE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLKEVVEEA)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLKEVVEEAEN)
Or the compound of following structure:
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLLKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLLKKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLLKEVVEE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLLEKKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLLKEKKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLLKEVVEKEVVE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLLKEVVEEAE)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLLKEVVEEA)
PEG-CH 2cH 2-X-(CH 2) mcO-ring (CLLKEVVEEAEN)
Wherein, X=O, NH or NHCO; M=0-6.
Another aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, in the compound that wherein (VIII) organizes, the carboxyl in any site is modified through PEG, obtain following compound shown in (IX), wherein said amino comprises the side chain carboxyl group of C-Amino End Group and natural amino acid or alpha-non-natural amino acid:
[PEG-X-CO] z-ring (X 1-Y 1-Y 2-X 2)
[PEG-X-CO] z-ring (X 1-X 2-Y 1-Y 2-X 3)
[PEG-X-CO] z-ring (X 1-X 2-X 3-Y 1-Y 2-X 4)
[PEG-X-CO] z-ring (X 1-X 2-Y 1-Y 2-X 3-X 4)
[PEG-X-CO] z-ring (X 1-X 2-X 3-y 1-Y 2-X 4-X 5)
[PEG-X-CO] z-ring (X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3)
[PEG-X-CO] z-ring (X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3)
[PEG-X-CO] z-ring (X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5)
[PEG-X-CO] z-ring (X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6)
[PEG-X-CO] z-ring (X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-y 4)
[PEG-X-CO] z-ring (X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-y 4)
[PEG-X-CO] z-ring (X 1-X 2-X 3-X 4-Y 1-Y 2-X 5)
[PEG-X-CO] z-ring (X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6)
[PEG-X-CO] z-ring (X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7)
[PEG-X-CO] z-ring (X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
[PEG-X-CO] z-ring (X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y3-Y4-X 7)
[PEG-X-CO] z-ring (Cys-X 1-Y 1-Y 2-X 2)
[PEG-X-CO] z-ring (Cys-X 1-X 2-Y 1-Y 2-X 3)
[PEG-X-CO] z-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4)
[PEG-X-CO] z-ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4)
[PEG-X-CO] z-ring (Cys-X 1-X 2-X 3-y 1-Y 2-X 4-X 5)
[PEG-X-CO] z-ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3)
[PEG-X-CO] z-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3)
[PEG-X-CO] z-ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5)
[PEG-X-CO] z-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6)
[PEG-X-CO] z-ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-y 4)
[PEG-X-CO] z-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4)
[PEG-X-CO] z-ring (Cys-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5)
[PEG-X-CO] z-ring (Cys-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6)
[PEG-X-CO] z-ring (Cys-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7)
[PEG-X-CO] z-ring (Cys-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
Or following structure:
[PEG-X-CO] z-ring (Z 1-L n-X 1-Y 1-Y 2-X 2)
[PEG-X-CO] z-ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 3)
[PEG-X-CO] z-ring (Z 1-L n-X 1-X 2-X 3-Y 1-Y 2-X 4)
[PEG-X-CO] z-ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 3-X 4)
[PEG-X-CO] z-ring (Z 1-L n-X 1-X 2-X 3-y 1-Y 2-X 4-X 5)
[PEG-X-CO] z-ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3)
[PEG-X-CO] z-ring (Z 1-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-Y 4)
[PEG-X-CO] z-ring (Z 1-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-y 4)
[PEG-X-CO] z-ring (Z 1-L n-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5)
[PEG-X-CO] z-ring (Z 1-L n-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6)
[PEG-X-CO] z-ring (Z 1-L n-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7)
[PEG-X-CO] z-ring (Z 1-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
[PEG-X-CO] z-ring (Z 1-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y3-Y4-X 7) (IX)
Wherein, X 1, X 2, Y 1, Y 2, X 3, X 4, X 5, X 6, X 7, Y 1, Y 2, Y 3, Y 4definition as shown in claim 1, Z 1, L ndefinition as shown in aforementioned; X=O or NH, Z=1-3.
Another aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, is selected from:
[PEG-X-CO] z-ring (EVVE)
[PEG-X-CO] zring (KEVVE)
[PEG-X-CO] zring (KKEVVE)
[PEG-X-CO] zring (KEVVEE)
[PEG-X-CO] zring (EKKEVVE)
[PEG-X-CO] zring (KEKKEVVE)
[PEG-X-CO] zring (KEVVEKEVVE)
[PEG-X-CO] zring (KEVVEEAE)
[PEG-X-CO] zring (KEVVEEA)
[PEG-X-CO] zring (KEVVEEAEN)
[PEG-X-CO] z-ring (CEVVE)
[PEG-X-CO] zring (CKEVVE)
[PEG-X-CO] zring (CKKEVVE)
[PEG-X-CO] zring (CKEVVEE)
[PEG-X-CO] zring (CEKKEVVE)
[PEG-X-CO] zring (CKEKKEVVE)
[PEG-X-CO] zring (CKEVVEKEVVE)
[PEG-X-CO] zring (CKEVVEEAE)
[PEG-X-CO] zring (CKEVVEEA)
[PEG-X-CO] zring (CKEVVEEAEN)
Or following structure:
[PEG-X-CO] zring (CLEVVE)
[PEG-X-CO] zring (CLKEVVE)
[PEG-X-CO] zring (CLKKEVVE)
[PEG-X-CO] zring (CLKEVVEE)
[PEG-X-CO] zring (CLEKKEVVE)
[PEG-X-CO] zring (CLKEKKEVVE)
[PEG-X-CO] zring (CLKEVVEKEVVE)
[PEG-X-CO] zring (CLKEVVEEAE)
[PEG-X-CO] zring (CLKEVVEEA)
[PEG-X-CO] zring (CLKEVVEEAEN)
Or the compound of following structure:
[PEG-X-CO] zring (CLLEVVE)
[PEG-X-CO] zring (CLLKEVVE)
[PEG-X-CO] zring (CLLKKEVVE)
[PEG-X-CO] zring (CLLKEVVEE)
[PEG-X-CO] zring (CLLEKKEVVE)
[PEG-X-CO] zring (CLLKEKKEVVE)
[PEG-X-CO] zring (CLLKEVVEKEVVE)
[PEG-X-CO] zring (CLLKEVVEEAE)
[PEG-X-CO] zring (CLLKEVVEEA)
[PEG-X-CO] zring (CLLKEVVEEAEN).
Another aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, wherein (VIII) compound of organizing is after in peptide sequence or existing cysteine position or sequence, halfcystine is introduced in optional position, through being selected from PEG-MAL, the method of PEG-VS and PEG-IODS is modified, obtain the compound that (X) as follows organizes, wherein PEG can be chain, starlike, ring-type, multi-arm type structure:
PEG-M-ring (Cys-X 1-Y 1-Y 2-X 2)
PEG-M-ring (Cys-X 1-X 2-Y 1-Y 2-X 3)
PEG-M-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4)
PEG-M-ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4)
PEG-M-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5)
PEG-M-ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3)
PEG-M-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3)
PEG-M-ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5)
PEG-M-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6)
PEG-M-ring (Cys-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-y 4)
PEG-M-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-Y 4)
PEG-M-ring (Cys-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5)
PEG-M-ring (Cys-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6)
PEG-M-ring (Cys-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7)
PEG-M-ring (Cys-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
PEG-M-ring (Cys-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
Also can be following structure:
PEG-M-ring (Cys-L n-X 1-Y 1-Y 2-X 2)
PEG-M-ring (Cys-L n-X 1-X 2-Y 1-Y 2-X 3)
PEG-M-ring (Cys-L n-X 1-X 2-X 3-Y 1-Y 2-X 4)
PEG-M-ring (Cys-L n-X 1-X 2-Y 1-Y 2-X 3-X 4)
PEG-M-ring (Cys-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5)
PEG-M-ring (Cys-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3)
PEG-M-ring (Cys-L n-X 1-X 2-Y 1-Y 2-X 3-X 4-Y 3-X 5-Y 4)
PEG-M-ring (Cys-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-Y 3-X 6-y 4)
PEG-M-ring (Cys-L n-X 1-X 2-X 3-X 4-X 5-X 6-Y 1-Y 2-X 7)
PEG-M-ring (Cys-X 1-X 2-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
PEG-M-ring (Cys-L n-X 1-X 2-X 3-Y 1-Y 2-X 4-X 5-X 6-Y 3-Y 4-X 7)
PEG-M-ring (Cys-L n-X 1-X 2-X 3-X 4-Y 1-Y 2-X 5)
PEG-M-ring (Cys-L n-X 1-X 2-X 3-X 4-X 5-Y 1-Y 2-X 6) (X)
Wherein,
Wherein X 1, Z 1can be halfcystine, be connected with M group covalency by side chain sulfur atom; X 1, X 2, Y 1, Y 2, X 3, X 4, X 5, X 6, X 7, Y 1, Y 2, Y 3, Y 4definition as shown in aforementioned, Z 1, L ndefinition as shown in aforementioned.
Another aspect of the present invention relates to tetrapeptide to a ten as described below peptide derivant and steric isomer thereof, and it is selected from:
[PEG-M]-ring (CEVVE)
[PEG-M]-ring (CKEVVE)
[PEG-M]-ring (CKKEVVE)
[PEG-M]-ring (CKEVVEE)
[PEG-M]-ring (CEKKEVVE)
[PEG-M]-ring (CKEKKEVVE)
[PEG-M] ring (CKEVVEKEVVE)
[PEG-M] ring (CKEVVEEAE)
[PEG-M]-ring (CKEVVEEA)
[PEG-M]-ring (CKEVVEEAEN)
Or following structure:
[PEG-M]-ring (CLEVVE)
[PEG-M]-ring (CLKEVVE)
[PEG-M]-ring (CLKKEVVE)
[PEG-M]-ring (CLKEVVEE)
[PEG-M]-ring (CLEKKEVVE)
[PEG-M]-ring (CLKEKKEVVE)
[PEG-M]-ring (CLKEVVEKEVVE)
[PEG-M]-ring (CLKEVVEEAE)
[PEG-M]-ring (CLKEVVEEA)
[PEG-M]-ring (CLKEVVEEAEN)
Or the compound of following structure:
[PEG-M]-ring (CLLEVVE)
[PEG-M]-ring (CLLKEVVE)
[PEG-M]-ring (CLLKKEVVE)
[PEG-M]-ring (CLLKEVVEE)
[PEG-M] ring (CLLEKKEVVE)
[PEG-M]-ring (CLLKEKKEVVE)
[PEG-M]-ring (CLLKEVVEKEVVE)
[PEG-M]-ring (CLLKEVVEEAE)
[PEG-M]-ring (CLLKEVVEEA)
[PEG-M]-ring (CLLKEVVEEAEN).
The invention still further relates at least one be selected from above-mentioned (I), (II), (III), the compound organized of (IV), (V), (VI), (VII), (VIII), (IX), (X) or its any combination purposes in for the preparation of the medicine treating and/or preventing immune deficiency or hypoimmunity diseases related.
The invention still further relates to pharmaceutical composition, be selected from above-mentioned (I), (II), (III) comprising at least one, compound that (IV), (V), (VI), (VII), (VIII), (IX), (X) organize, and pharmaceutical carrier or vehicle.
The invention still further relates to preparation above-mentioned (I), (II), (III), the method for compound organized of (IV), (V), (VI), (VII), (VIII), (IX), (X), it comprises with known solution method and Solid phase synthesis.
The present invention's PEG-OH structure used is: RO (CH 2cH 2o) NCH 2cH 2oH, wherein R=H or CH 3, molecular-weight average can be bought as commercial reagents by the PEG-OH of hundreds of to several ten thousand, PEG-NH 2, PEG-NHCOCH 2cH 2cOOH can be obtained by following approach reaction:
Concrete, first introduce the amino acid of amino, carboxyl or preparation PEGization in one end of PEG, then be coupled in peptide sequence with solution method or solid-phase synthesis and go, the N-Amino End Group to polypeptide can be realized, C-end carboxyl, Lys side-chain amino group, the modification of Asp or Glu side chain carboxyl group, by PEG-NH 2with maleic anhydride, vinyl sulfoxide, iodo acetic acid anhydride reactant can obtain PEG-MAL, PEG-VS, PEG-IODO.
WangResin, MBHAResin, DCC, HOBt, HBTU, BOP, Fmoc amino acid involved in the present invention, BOC protected amino acid, the reagent such as TFA, DIEA, mPEG-OH, NMM all obtain by buying.All alpha-non-natural amino acids synthesize by this laboratory and split.
According to the present invention, the disease relevant with immune deficiency or immunologic hypofunction has: hepatitis B, hepatitis C, nonsmall-cell lung cancer, melanoma, acquired immune deficiency syndrome (AIDS) etc.
According to the present invention, term used, as T α 1 (17-27), refers to and holds C-to hold the sequence of corresponding position from N-in T α 1.
The T α 1 active fragments derivative that illustrated (I)-(X) organizes and steric isomer thereof or the pharmaceutical composition containing it can administrations in a unit, route of administration can be enteron aisle or non-bowel, as oral, muscle, subcutaneous, nasal cavity, oral mucosa, skin, peritonaeum or rectum etc.
Form of administration is tablet, capsule, dripping pill, aerosol, pill, pulvis, solution, suspensoid, emulsion, granule, liposome, transdermal agent, buccal tablet, suppository, lyophilized injectable powder etc. such as.Can be ordinary preparation, sustained release preparation, controlled release preparation and various particulate delivery system.
In order to unit dosage forms for administration is made tablet, various carrier well known in the art can be widely used.About the example of carrier, comprise such as thinner and absorption agent, as starch, dextrin, lactose, N.F,USP MANNITOL, sucrose, glucose, calcium sulfate, sodium-chlor, urea, calcium carbonate, white bole, Microcrystalline Cellulose, pure aluminium silicate etc.; Wetting agent and tackiness agent, as water, glycerine, polyoxyethylene glycol, ethanol, propyl alcohol, starch slurry, dextrin, syrup, honey, glucose solution, mucialga of arabic gummy, gelatine size, Xylo-Mucine, lac, methylcellulose gum, potassiumphosphate, polyvinylpyrrolidone etc.; Disintegrating agent, as dry starch, alginates, agar powder, laminaran, sodium bicarbonate and Citric Acid, calcium carbonate, polyoxyethylene sorbitol fatty acid ester, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.; Disintegration inhibitor, as sucrose, Tristearoylglycerol, theobroma oil, hydrogenation wet goods; Absorption enhancer, as quaternary ammonium salt, sodium lauryl sulphate etc.; Lubricant, as talcum powder, silicon-dioxide, W-Gum, stearate, boric acid, whiteruss, polyoxyethylene glycol etc.Tablet can also be made coating tablet further, as sugar coated tablet, thin membrane coated tablet, ECT, or double-layer tablets and multilayer tablet.In order to administration unit is made pill, various carrier well known in the art can be widely used.Example about carrier is, such as thinner and absorption agent, as glucose, lactose, starch, theobroma oil, hydrogenated vegetable oil, polyvinylpyrrolidone, Gelucire, kaolin, talcum powder etc.; Tackiness agent is as gum arabic, tragacanth gum, gelatin, ethanol, honey, rice paste or batter etc.; Disintegrating agent, as agar powder, dry starch, alginates, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.In order to administration unit is made suppository, various carrier well known in the art can be widely used.Example about carrier is, the ester, gelatin, semi-synthetic glyceryl ester etc. of such as polyoxyethylene glycol, Yelkin TTS, theobroma oil, higher alcohols, higher alcohols.In order to administration unit is made capsule, effective constituent formula (I)-(X) T α 1 active fragments derivative or its steric isomer are mixed with above-mentioned various carriers, and the mixture obtained thus is placed in hard capsule or soft capsule.Also tripeptide derivative effective constituent (I)-(X) can organized or its steric isomer make microcapsule, are suspended in aqueous medium and make suspensoid, also can load in hard capsule or make injection application.In order to administration unit is made injection preparation, as solution, emulsion, lyophilized injectable powder and suspensoid, all thinners that this area is conventional can be used, as the isooctadecanol of water, ethanol, polyoxyethylene glycol, 1,3-PD, ethoxylation, the isooctadecanol, Polyoxyethylene Sorbitol Fatty Acid Esters etc. of polyoxy base.In addition, in order to prepare isotonic injection liquid, appropriate sodium-chlor, glucose or glycerine can be added in injection preparation, in addition, conventional solubility promoter, buffer reagent, pH adjusting agent etc. can also be added.
In addition, as needs, also tinting material, sanitas, spices, correctives, sweeting agent or other material can be added in pharmaceutical preparation.
The compound that the present invention (I)-(X) organizes, the dosage of derivative or its steric isomer depends on many factors, such as to prevent or treat (I)-(X) character of T α 1 active fragments derivative disease of organizing and severity, the sex of patient or animal, age, body weight and individual reaction, particular compound used, route of administration and administration number of times etc.Above-mentioned dosage can single dose form or be divided into several, and such as two, at, four dosage forms for administration.The abbreviation used in the present invention has implication below:
In the present invention, all amino acid configuration, except being labeled as D-type, are L-type.
Embodiment
The following example represents illustrative embodiment of the present invention, but does not mean that the present invention is subject to the restriction of this embodiment.
Embodiment solid-phase synthesized carrier Wang resin used is buied from ACT company, and DCC, HOBt, HBTU, Fmoc protected amino acid is buied by Shanghai gill biochemistry, and TFA is ACROS Products, molecular-weight average be 2000 PEG-OH be Sigma Products.
Embodiment 1
BocNHCH 2cH 2oCH 2the synthesis of COOH:
● BocNHCH 2cH 2the synthesis of OH
Water 30ml is added in 250ml reactor, NH 2cH 2cH 2oH6.1g (100mmol) adds in reactor, is cooled to 0 DEG C.Under condition of ice bath, (Boc) 2o21.82g (100mmol) is dissolved in 30ml dioxane and joins in reactor, and control temperature is below 0 DEG C, and drip 1mol/LnaOH solution 100ml, 40min drips, and naturally rises to room temperature, continues reaction 12 hours, (Boc) 2o disappears, stopped reaction, and some plate detects, and reaction solution is extracted with ethyl acetate three times, washs by saturated NaCl solution, uses anhydrous Na 2sO4 is dry, and evaporated under reduced pressure, obtains 15.6g colorless oil, yield 97.2%.
● BocNHCH 2cH 2oCH 2the synthesis of COOH
Add in 500ml there-necked flask and heavily steam DMF350ml, BocNH 2cH 2oH10.54g (0.1mol), add NaH (52%) 8.15g (0.2mol), to solution, bubble-free is emerged, and then drips BrCH 2the DMF solution of COOH13.65g (0.15mmol), stirring at room temperature 20 hours, processes, pressure reducing and steaming DMF, with suitable quantity of water dissolve, by appropriate extracted with diethyl ether twice, aqueous phase by saturated sodium sulfate adjust ph between 2-3, be extracted with ethyl acetate aqueous phase three times, then wash by saturated NaCl solution, finally use anhydrous sodium sulfate drying, evaporated under reduced pressure, obtain 5.51g light yellow oil, yield 38%.
Embodiment 2:
Cys-Glu-Val-Val-Glu-SCH 2cH 2cONH 2synthesis:
(charge capacity is 0.57mmol/g to 350mgMBHA resin, 0.2mmol), be suspended in DMF, add 0.12gHOBt successively, 150 μ L3-thiohydracrylic acids (1.2mmol), 160mgDCC, adding a small amount of DCM dissolves fully, at room temperature magnetic agitation 4 hours, respectively washes twice with DMF, DCM, MeOH, DCM, each two minutes.Triketohydrindene hydrate inspection is negative.Then 50mg acthiol-J hydrochloride (0.2mmol) is used, 100mg (0.2mmol) triphenyl phosphorus and 120 μ lDIEA (0.2mmol) mixing are dissolved in DMF, add after five minutes in resin, room temperature reaction two hours, with DMF, DCM, MeOH, each twice of DCM washing, the amino acid of the BOC protection of 4 times amount, BOP, the DIEA mixing of 6 times amount, be dissolved in DMF, after five minutes, mixture joins in reactor, room temperature reaction 2 hours, with DMF, DCM, MeOH, DCM washs, detect with Ellman experiment qualitatively, negative solution resin is colourless or faint yellow, if in orange, then to reduce by half reaction one hour with above-mentioned condensation dosage, application BOC/Bzl solid phase condensation system progressively extends peptide chain from C end, with 4NHCl/DIOX (0.05% indoles) deprotection, condensation reaction reacts 1 hour with the DCC/HOBt of the BOC-AA of 2 times amount and 2 times amount, often walk condensation all to detect with triketohydrindene hydrate experiment, if for positive, then use the BOC-AA of 4 times amount, the DIEA of BOP and 6 times amount reacts half hour, after peptide chain end of synthesis, linear peptides thioesters HF/P-thiocresol/m-cresol (93/2/5, v/v/v), cut from resin after 1.5 hours 0 DEG C of reaction, HF drains the freezing anhydrous diethyl ether washing of rear sodium silk drying for several times, Glacial acetic acid with 10% extracts thick peptide, freeze-drying is preserved.
Embodiment three: the synthesis of ring (CEVVE):
(charge capacity is 0.57mmol/g to 350mgMBHA resin, 0.2mmol), be suspended in DMF, add 0.12gHOBt successively, 150 μ L3-thiohydracrylic acids (1.2mmol), 160mgDCC, adding a small amount of DCM dissolves fully, at room temperature magnetic agitation 4 hours, respectively washes twice with DMF, DCM, MeOH, DCM, each two minutes.Triketohydrindene hydrate inspection is negative.Then 50mg acthiol-J hydrochloride (0.2mmol) is used, 100mg (0.2mmol) triphenyl phosphorus and 120 μ lDIEA (0.2mmol) mixing are dissolved in DMF, add after five minutes in resin, room temperature reaction two hours, with DMF, DCM, MeOH, each twice of DCM washing, the amino acid of the BOC protection of 4 times amount, BOP, the DIEA mixing of 6 times amount, be dissolved in DMF, after five minutes, mixture joins in reactor, room temperature reaction 2 hours, with DMF, DCM, MeOH, DCM washs, detect with Ellman experiment qualitatively, negative solution resin is colourless or faint yellow, if in orange, then to reduce by half reaction one hour with above-mentioned condensation dosage, application BOC/BZL solid phase condensation system progressively extends peptide chain from C end, with 4NHCl/DIOX (0.05% indoles) deprotection, condensation reaction reacts 1 hour with the DCC/HOBt of the BOC-AA of 2 times amount and 2 times amount, often walk condensation all to detect with triketohydrindene hydrate experiment, if for positive, then use the BOC-AA of 4 times amount, the DIEA of BOP and 6 times amount reacts half hour, after peptide chain end of synthesis, linear peptides thioesters HF/P-thiocresol/m-cresol (93/2/5, v/v/v), cut from resin after 1.5 hours 0 DEG C of reaction, HF drains the freezing anhydrous diethyl ether washing of rear sodium silk drying for several times, Glacial acetic acid with 10% extracts thick peptide, freeze-drying is preserved.Also the buffered soln extracting directly linear peptides thioesters of cyclization can be carried out ring-closure reaction.If peptide thioesters solubleness is bad, it is also conceivable to use CH 3cN/H 2o extracts thick peptide, and then decompression pumps CH 3after CN, freeze-drying is preserved.
Linear peptides thioesters carries out ring-closure reaction after purifying with gel filtration chromatography preliminary purification or HPLC, reacts at NaHCO 3the aqueous solution in carry out, if solubleness is bad suitably can add CH 3the concentration of CN, control ph is at 7.3-7.6, and add the oxidation that TCEP prevents sulfydryl, react and monitored by HPLC, react and completed in 4 hours, product ESI-MS identifies.
(the synthesis of CKEVVEE of embodiment four: PEG-MAL-ring
1mPEG 5000the synthesis of-MAL
Weighing m PEG 5000-OH20g (4mmol) is placed in 250ml reaction flask, adds 100mlCH 2cl 2, after dissolution of solid, add 3.0mlEt again 3n (20mmmol) and 3.8gTs-Cl (20mmol), stirring at room temperature is reacted.TLC (CH 3oH: CH 2cl 2=1: 6), after monitoring reacts completely, rotary evaporation, except desolventizing, adds 100ml anhydrous diethyl ether and is settled out solid, then use dehydrated alcohol recrystallization twice, obtain 15.2gmPEG 5000-OTs, yield 70%.
By 15gmPEG 5000-OTs (3mmol) is dissolved in 30mlDMF, adds 1.68g (18mmol) potassium phthalimide, at N 2under protection, 120 DEG C are reacted 4 hours.Pressure reducing and steaming solvent, is dissolved in 50ml dehydrated alcohol by resistates, add 2.0ml hydrazine hydrate, back flow reaction 4 hours.Resistates, except desolventizing, is dissolved in CH by rotary evaporation 2cl 2, cross and filter insolubles, then by filtrate rotary evaporation except desolventizing, be settled out solid with anhydrous diethyl ether, then with dehydrated alcohol-Diethyl ether recrystallization, obtain 12.5gmPEG 5000-NH 2, yield 83%.
By 1.25gmPEG 5000-NH 2be dissolved in 10ml dioxane, add maleic anhydride 0.1g, 80 DEG C of stirring reaction 30min.Pressure reducing and steaming solvent, adds 50ml anhydrous diethyl ether, and cooling is settled out solid, and filter collection solid, obtains 1.10g after drying.Gained solid is dissolved in 15ml diacetyl oxide, adds 0.5g sodium acetate, 100 DEG C of stirring reaction 45min.Pressure reducing and steaming solvent, dissolved by residue from dichloromethane, elimination insolubles, adds proper amount of active carbon in filtrate, place 30min, filtering gac, is concentrated into dry by filtrate, add anhydrous diethyl ether, be settled out solid, after filter collection, drying, obtain light yellow solid 0.61gmPEG 5000-MAL, yield 48%.RP-HPLC analyzes: t r=36.79min.
Chromatographic condition: Waters600HPLC
Chromatographic column: InersilODS-3 (10 × 250mm) ultraviolet detection wavelength: 215nm
Moving phase: A-0.05%TFA/H 2oB-0.05%TFA/70%CH 3cN/H 2o
Flow velocity: 2.0mL/min
Gradient (linearly): t (min) A%B%
01000
51000
350100
400100
451000
(charge capacity is 0.57mmol/g to 350mgMBHA resin, 0.2mmol), be suspended in DMF, add 0.12gHOBt successively, 150 μ L3-thiohydracrylic acids (1.2mmol), 160mgDCC, adding a small amount of DCM dissolves fully, at room temperature magnetic agitation 4 hours, respectively washes twice with DMF, DCM, MeOH, DCM, each two minutes.Triketohydrindene hydrate inspection is negative.Then 50mg acthiol-J hydrochloride (0.2mmol) is used, 100mg (0.2mmol) triphenyl phosphorus and 120 μ lDIEA (0.2mmol) mixing are dissolved in DMF, add after five minutes in resin, room temperature reaction two hours, with DMF, DCM, MeOH, each twice of DCM washing, the amino acid of the BOC protection of 4 times amount, BOP, the DIEA mixing of 6 times amount, be dissolved in DMF, after five minutes, mixture joins in reactor, room temperature reaction 2 hours, with DMF, DCM, MeOH, DCM washs, detect with Ellman experiment qualitatively, negative solution resin is colourless or faint yellow, if in orange, then to reduce by half reaction one hour with above-mentioned condensation dosage, application BOC/Bzl solid phase condensation system progressively extends peptide chain from C end, with 4NHCl/DIOX (0.05% indoles) deprotection, condensation reaction reacts 1 hour with the DCC/HOBt of the BOC-AA of 2 times amount and 2 times amount, often walk condensation all to detect with triketohydrindene hydrate experiment, if for positive, then use the BOC-AA of 4 times amount, the DIEA of BOP and 6 times amount reacts half hour, after peptide chain end of synthesis, linear peptides thioesters HF/P-thiocresol/m-cresol (93/2/5, v/v/v), cut from resin after 1.5 hours 0 DEG C of reaction, HF drains the freezing anhydrous diethyl ether washing of rear sodium silk drying for several times, Glacial acetic acid with 10% extracts thick peptide, freeze-drying is preserved.
2, the synthesis of ring (CEVVE):
(charge capacity is 0.57mmol/g to 350mgMBHA resin, 0.2mmol), be suspended in DMF, add 0.12gHOBt successively, 150 μ L3-thiohydracrylic acids (1.2mmol), 160mgDCC, adding a small amount of DCM dissolves fully, at room temperature magnetic agitation 4 hours, respectively washes twice with DMF, DCM, MeOH, DCM, each two minutes.Triketohydrindene hydrate inspection is negative.Then 50mg acthiol-J hydrochloride (0.2mmol) is used, 100mg (0.2mmol) triphenyl phosphorus and 120 μ lDIEA (0.2mmol) mixing are dissolved in DMF, add after five minutes in resin, room temperature reaction two hours, with DMF, DCM, MeOH, each twice of DCM washing, the amino acid of the BOC protection of 4 times amount, BOP, the DIEA mixing of 6 times amount, be dissolved in DMF, after five minutes, mixture joins in reactor, room temperature reaction 2 hours, with DMF, DCM, MeOH, DCM washs, detect with Ellman experiment qualitatively, negative solution resin is colourless or faint yellow, if in orange, then to reduce by half reaction one hour with above-mentioned condensation dosage, application BOC/BZL solid phase condensation system progressively extends peptide chain from C end, with 4NHCl/DIOX (0.05% indoles) deprotection, condensation reaction reacts 1 hour with the DCC/HOBt of the BOC-AA of 2 times amount and 2 times amount, often walk condensation all to detect with triketohydrindene hydrate experiment, if for positive, then use the BOC-AA of 4 times amount, the DIEA of BOP and 6 times amount reacts half hour, after peptide chain end of synthesis, linear peptides thioesters HF/P-thiocresol/m-cresol (93/2/5, v/v/v), cut from resin after 1.5 hours 0 DEG C of reaction, HF drains the freezing anhydrous diethyl ether washing of rear sodium silk drying for several times, Glacial acetic acid with 10% extracts thick peptide, freeze-drying is preserved.Also the buffered soln extracting directly linear peptides thioesters of cyclization can be carried out ring-closure reaction.If peptide thioesters solubleness is bad, it is also conceivable to use CH 3cN/H 2o extracts thick peptide, and then decompression pumps CH 3after CN, freeze-drying is preserved.
Linear peptides thioesters carries out ring-closure reaction after purifying with gel filtration chromatography preliminary purification or HPLC, reacts at NaHCO 3the aqueous solution in carry out, if solubleness is bad suitably can add CH 3the concentration of CN, control ph is at 7.3-7.6, and add the oxidation that TCEP prevents sulfydryl, react and monitored by HPLC, react and completed in 4 hours, product ESI-MS identifies.
3, the PEG-MAL of cyclic peptide modifies
By soluble in water for the ring (CEVVE) after RP-HPLC purifying, adjust pH to 8 with sodium bicarbonate, add the mPEG of 3 times of equivalents 5000-MAL, room temperature reaction, monitors reaction process and separated product with RP-HPLC, and reaction generally completed in four hours.Cys (mPEG 5000-MAL)-T α 1(17-24) analyze through MALDI-TOF-MS, near 6173, have a series of peak, adjacent two peak mass difference about 44, have the typical structural characteristics of polyoxyethylene glycol.By with the mPEG-NH not connecting cyclic peptide molecule 2relatively, deduct the formula weight of the maleoyl imido as activating functional group, the molecular weight of its value fundamental sum cyclic peptide is corresponding, proves that the mPEGization successfully achieving cyclic peptide compound is modified.
Embodiment 5: 3h-tdR incorporation methods detects reacts the increment of mouse spleen lymphocyte:
The preparation of splenocyte suspension: aseptic taking-up spleen, grinds mouse spleen with frosted glass plate, make splenocyte suspension.After splitting erythrocyte, wash three times, counting (viable cell is more than 95%).With the RPMI1640 nutrient solution containing 10%FBS, splenocyte concentration is adjusted to 4 × 10 6cell/ml.This experiment adds ConA (concanavalin A) 5ug/ml inducer T lymphocyte propagation.With 3the propagation of H-TdR incorporation methods quantitative assay cell.At 37C, 5%CO 2cultivate 48 hours in incubator.7-8 hour before end is cultivated, every hole adds 25 μ l 3h-thymidylic acid is (containing 2 × 10 4bq).After cultivation terminates, with cell collector (HARVESTER, TOMTEC) collecting cell on glass fibre membrane, with liquid dodge numeration instrument (MicroBetaTrilux, PerkinElmer) detect [ 3h] incorporation of-thymidine in DNA, represent cell proliferative conditions with cpm value.Concrete detected result is see table 1.
Result evaluation: lymphocytic proliferative response adopts detected sample cpm value to deduct positive control sample cpm value, then removes positive control sample cpm value, is labeled as+strengthens/% of-suppression.In view of test sample does not have the effect of non-specific cellular toxicity, result queue be+(enhancings)/-(suppression) more than 15% (according to during P value usual more than 10% also can), have activity with regard to expression sample.
Table 1T α 1and the T lymphproliferation response result of analogue
Cpm value represents cell proliferative conditions, and SD is the standard deviation of data, and P value represents that experimental data and ConA control group data carry out the probability having indifference of statistical study between this, and P < 0.05 represents has significant difference between two groups of data.L＝-NHCH 2CH 2OCH 2COOH-。

Claims (3)

1. cyclic peptide or its PEG modifier, wherein said cyclic peptide is the compound being selected from following structure:
Ring (CKEVVEKEVVE)
Ring (CKEVVEEA).
2. cyclic peptide according to claim 1 or its PEG modifier are for the preparation of the purposes of medicine treating and/or preventing immune deficiency, immunologic hypofunction relative disease.
3. cyclic peptide according to claim 1 or its PEG modifier are for the preparation of the purposes of healthcare products.
CN201110034234.4A 2004-12-01 2004-12-01 Thymosin α1 active fragments cyclic peptide analogue and polyethylene glycol derivative thereof Expired - Fee Related CN102174084B (en)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN2004100964592A CN1781933B (en) 2004-12-01 2004-12-01 Thymosin alpha 1 active segment cyclicpeptide analogue and its poly glycol derivative

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
CN2004100964592A Division CN1781933B (en) 2004-12-01 2004-12-01 Thymosin alpha 1 active segment cyclicpeptide analogue and its poly glycol derivative

Publications (2)

Publication Number Publication Date
CN102174084A CN102174084A (en) 2011-09-07
CN102174084B true CN102174084B (en) 2015-11-18

Family

ID=36772597

Family Applications (2)

Application Number Title Priority Date Filing Date
CN2004100964592A Expired - Fee Related CN1781933B (en) 2004-12-01 2004-12-01 Thymosin alpha 1 active segment cyclicpeptide analogue and its poly glycol derivative
CN201110034234.4A Expired - Fee Related CN102174084B (en) 2004-12-01 2004-12-01 Thymosin α1 active fragments cyclic peptide analogue and polyethylene glycol derivative thereof

Family Applications Before (1)

Application Number Title Priority Date Filing Date
CN2004100964592A Expired - Fee Related CN1781933B (en) 2004-12-01 2004-12-01 Thymosin alpha 1 active segment cyclicpeptide analogue and its poly glycol derivative

Country Status (1)

Country Link
CN (2) CN1781933B (en)

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1781933B (en) * 2004-12-01 2011-07-20 中国人民解放军军事医学科学院毒物药物研究所 Thymosin alpha 1 active segment cyclicpeptide analogue and its poly glycol derivative
CN102746385B (en) * 2004-12-01 2014-07-09 中国人民解放军军事医学科学院毒物药物研究所 Thymosin alpha 1 active fragment cyclopeptide analogue and polyethylene glycol derivative thereof
CN1927879B (en) * 2006-09-25 2010-11-24 吉林大学 Thymopentapeptide active isomer and application thereof in pharmaceutical preparation
EP2168602A4 (en) * 2007-06-12 2014-10-29 Inst Pharm & Toxicology Amms Site-specific pegylated linear salmon calcitonin derivatives
CN102796168B (en) * 2011-05-27 2014-02-26 首都医科大学 Compounds with thrombolytic activity and preparation method and application thereof
WO2017004811A1 (en) * 2015-07-08 2017-01-12 中国食品发酵工业研究院 Active peptide for inhibiting ampa receptor and preparation method and use thereof
CN114173791A (en) * 2019-07-17 2022-03-11 波兹皮埃德公司 Boron-containing RHO kinase inhibitors
CN111454329A (en) * 2020-04-07 2020-07-28 哈尔滨师范大学 Isoindolinone bridged cyclic nonapeptide with anticancer activity and preparation method thereof
CN112315909B (en) * 2020-11-05 2023-02-17 南开大学 Functional polymer micelle PM-TLK and preparation method and application thereof

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4547489A (en) * 1984-06-11 1985-10-15 Ortho Pharmaceutical Corporation Conformationally restricted thymopentin-like compounds
US4910296A (en) * 1980-01-18 1990-03-20 Max-Planck-Gesellschaft Zur Foederung Der Wissenschaften E.V. Medicaments containing alpha 1 thymosin fragments and having an immunostimulant action, and fragments of alpha 1 thymosin
CN1532206A (en) * 2003-03-21 2004-09-29 中国人民解放军军事医学科学院毒物药 Active fragment of thymosin alphal and its polyethylene glycol derivatives
CN1781933B (en) * 2004-12-01 2011-07-20 中国人民解放军军事医学科学院毒物药物研究所 Thymosin alpha 1 active segment cyclicpeptide analogue and its poly glycol derivative

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4910296A (en) * 1980-01-18 1990-03-20 Max-Planck-Gesellschaft Zur Foederung Der Wissenschaften E.V. Medicaments containing alpha 1 thymosin fragments and having an immunostimulant action, and fragments of alpha 1 thymosin
US4547489A (en) * 1984-06-11 1985-10-15 Ortho Pharmaceutical Corporation Conformationally restricted thymopentin-like compounds
CN1532206A (en) * 2003-03-21 2004-09-29 中国人民解放军军事医学科学院毒物药 Active fragment of thymosin alphal and its polyethylene glycol derivatives
CN1781933B (en) * 2004-12-01 2011-07-20 中国人民解放军军事医学科学院毒物药物研究所 Thymosin alpha 1 active segment cyclicpeptide analogue and its poly glycol derivative

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
L.C Thomas et al.Activity of synthetic thymosin .alpha.1 C-terminalpeptidesintheazathioprine E-rosette inhibition assay.《Biochemistry》.1982,第21卷(第18期), *
王淑彩等.免疫促进剂胸腺肽的研究进展.《畜牧与兽医》.2002,第34卷(第10期), *
蒋志龙等.生物活性环肽研究进展.《中国药物化学杂志》.2004,第14卷(第02期), *

Also Published As

Publication number Publication date
CN1781933A (en) 2006-06-07
CN102174084A (en) 2011-09-07
CN1781933B (en) 2011-07-20

Similar Documents

Publication Publication Date Title
JP3266311B2 (en) Novel polypeptide and anti-HIV agent using the same
CN100427502C (en) Antineoplastic oligopeptide and its preparation method and application
CN101104078B (en) Polypeptide and protein medicine polyethylene coupling compound
FI77875C (en) FOERFARANDE FOER FRAMSTAELLNING AV NYA TERAPEUTISKT ANVAENDBARA PEPTIDER.
EP2611825B1 (en) Solid phase synthesis of h(gly2)glp-2
CN106232616B (en) Amphipathic synthetic antibacterial peptide, its medical composition and its use
US6103697A (en) Peptides with immunomodulatory effects
CN102174084B (en) Thymosin α1 active fragments cyclic peptide analogue and polyethylene glycol derivative thereof
RU2073685C1 (en) Polypeptides or their salts, pharmaceutical composition showing antiviral activity with respect to human immunodeficiency virus
CN104231067B (en) Erythropoietin mimetic peptide chemical dimer and application thereof
CA2020838C (en) Hemoregulatory peptides
WO2002020561A1 (en) Novel polypeptides and anti-hiv drugs containing the same
HU208027B (en) Process for producing peptide compounds and pharmaceutical composition containing them as active components
CN103450341A (en) Deuterohemin-His-Peptide (DhHP-6) derivatives, and preparation methods and uses thereof
WO2017101144A1 (en) Adipose tissue targeting polypeptide, preparation method therefor and application thereof
CN102746385B (en) Thymosin alpha 1 active fragment cyclopeptide analogue and polyethylene glycol derivative thereof
US4021387A (en) Eosinophilotactic tetrapeptides
CN103965297A (en) Polypeptide as well as preparation method and application thereof
Thierry et al. Synthesis and activity of NAcSerAspLysPro analogs on cellular interactions between T-cell and erythrocytes in rosette formation
CN102250251A (en) Polyethylene glycol derivative of enkephalin analogue
US4058512A (en) Synthetic peptides having growth promoting activity
CN113880916B (en) Yak skin antioxidant polypeptide and preparation method and application thereof
Matsunaga et al. Synthesis and bioactivity studies of two isosteric acyclic analogues of melanin concentrating hormone
CN116801896A (en) BL-8040 composition
CN116621943A (en) Long-acting hepatitis virus entry inhibitor

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20151118

Termination date: 20161201

CF01 Termination of patent right due to non-payment of annual fee