CN1532207A - Polyethylene glycol derivatives of thymosin alphal - Google Patents

Polyethylene glycol derivatives of thymosin alphal Download PDF

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Publication number
CN1532207A
CN1532207A CNA031314988A CN03131498A CN1532207A CN 1532207 A CN1532207 A CN 1532207A CN A031314988 A CNA031314988 A CN A031314988A CN 03131498 A CN03131498 A CN 03131498A CN 1532207 A CN1532207 A CN 1532207A
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thr
asp
glu
ser
lys
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CN100372870C (en
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刘克良
王良友
刘尚义
吴萍
赵修南
董泄建
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Institute of Pharmacology and Toxicology of AMMS
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Abstract

The present invention relates to polyethylene glycol derivatives of thymosin alpha-1, their preparation process, the medicine composition containing them, and their application in the medicine for preventing and treating diseases related with immune deficiency and hypoimmunity, including hepatitis B, hepatitis C, malignant melanoma, non-small cell lung carcinoma, SARS, etc.

Description

Extrasin alpha 1Polyethylene glycol derivative
Invention field
The present invention relates to extrasin alpha 1Polyethylene glycol derivative, its preparation method, the pharmaceutical composition that contains them, and the purposes in the medicine of treatment or prevention and immune deficiency, immunologic hypofunction relative disease, comprise being included in hepatitis B hepatitis C, malignant melanoma, application in the treatment of the SARS that lung cancer in non-cellule type, coronavirus cause relative diseases such as (serious acute respiratory syndromes) or the prevention.
Background technology
Extrasin alpha 1(T α 1) be that the thymus gland excretory is a kind of important polypeptide, be a kind of at the lymphocytic immunostimulant of T, can promote the ripe and differentiation of T cell, and impel the multiple lymphokine of sophisticated T emiocytosis (as interleukin-2 and gamma-interferon etc.), can also promote the generation of interleukin-2 acceptor.T α 1Be made up of 28 amino-acid residues, molecular weight is 3108, and iso-electric point is 4.2, and the N-end contains the acetylize structure.T α 1Primary structure as follows:
1????????????????????????????????????10
Ac-Ser-Asp-Ala-Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-
20?????????????????????????????28
Leu-Lys-Glu-Lys-Lys-Glu-Val-Val-Glu-Glu-Ala-Glu-Asn-OH
T α 1The clinical treatment research that is used for immune deficiency or immune downtrod disease starts from the eighties in 20th century, has obtained effect preferably at present.When being used the treatment chronic hepatitis B when independent use or with Interferon, rabbit, T α 1It is a kind of safe and effective medicine.For many other diseases, comprise hepatitis C, nonsmall-cell lung cancer, melanoma and acquired immune deficiency syndrome (AIDS), T α 1Certain curative effect is all arranged.In addition, T α 1Also can be used as the vaccine adjuvant, strengthen the effect of influenza vaccines and hepatitis B vaccine.
The T α of present clinical use 1Be synthetic, cost an arm and a leg, dosage is big, the cycle is long.Therefore, to T α 1Carry out structural modification, increase its stability to enzyme or non-enzyme environment, it is very necessary prolonging action time and increasing biological activity.
Summary of the invention
The inventor has now found that T α after deliberation 1Or its fragment can prolong T α behind polyoxyethylene glycol (PEG) covalent modification 1Or its fragment transformation period in vivo, thereby keeping T α 1Or the bioactive while of its fragment, reduce T α 1Or its segmental consumption and prolonged T α 1Or action time in its segmental body.
The present invention relates to the T α shown in the formula (I) 1Or its segmental PEGization derivative:
PEG-M-Cys-T?????(I)
Figure A0313149800051
Or
Figure A0313149800052
Or
Wherein PEG is RO (CH 2CH 2O) n-CH 2CH 2, R=H or CH 3, n=5-1000.
Cys is a halfcystine, and it links to each other with M group covalency by its side chain thioether atom; Cys also links to each other or links to each other with the C-end carboxyl formation amido linkage of T with its amino with the amino formation of the N-end amido linkage of its carboxyl and T, and the Cys position comprises between the two ends of T sequence, any two adjacent amino acid and the amino acid of alternative optional position.
T represents natural T α 1Complete sequence or contain T α 1(17-24) any fragment of sequence.
Further aspect of the present invention relates to the pharmaceutical composition that contains at least a formula (I) compound and pharmaceutical carrier or vehicle.
The invention still further relates at least a formula (I) compound is used for preventing or the medicine purposes of treatment and immune deficiency or immunologic hypofunction relative disease in preparation, be included in hepatitis B, third liver, malignant melanoma, application in the treatment of the relative diseases such as SARS that lung cancer in non-cellule type, coronavirus cause or the prevention.
The invention still further relates to the method for preparation formula (I) compound, it comprises: with PEG-MAL, and PEG-VS or PEG-IODO and HS-Cys-T reaction.
According to the present invention, the abbreviation of Shi Yonging has following implication in the present invention:
T α 1-extrasin alpha 1,
The PEG-polyoxyethylene glycol,
PEG-MAL-dimaleoyl imino polyoxyethylene glycol,
PEG-VS-ethene sulfuryl polyoxyethylene glycol,
PEG-IODO-iodo-acetamide base polyoxyethylene glycol,
The Ala-L-Ala,
The Asn-l-asparagine,
The Asp-aspartic acid,
The Cys-halfcystine,
Glu-L-glutamic acid,
The Ile-Isoleucine,
Lys-Methionin,
The Leu-leucine,
The Ser-Serine,
The Thr-Threonine,
The Val-Xie Ansuan,
The Ac-ethanoyl,
The Boc-tertbutyloxycarbonyl,
The Fmoc-fluorenylmethyloxycarbonyl,
The DMF-dimethyl formamide,
The DCC-dicyclohexylcarbodiimide,
HBTU-2-(1H-1-hydroxybenzotriazole)-1,1,3,3-tetramethyl-urea phosphofluoric acid,
The HOBT-1-hydroxybenzotriazole,
The HOSu-N-N-Hydroxysuccinimide,
The NMM-N-methylmorpholine,
The TFA-trifluoroacetic acid,
The Ts-Cl-Tosyl chloride
The EDT-mercaptoethanol,
The RP-HPLC-RPLC,
The IFN-Interferon, rabbit.
All amino acid configurations are the L-type among the present invention except that being labeled as the D-type.
According to the present invention, molecular-weight average involved in the present invention can be used as commercialization reagent by the PEG-OH of hundreds of to several ten thousand and buys PEG-NH 2Can buy or obtain by following reaction:
Figure A0313149800071
With PEG-NH 2Get PEG-MAL, PEG-VS and PEG-IODO with maleic anhydride, vinyl chlorination sulfoxide, iodo acetic anhydride respectively.For example PEG-MAL obtains by following reaction:
Figure A0313149800072
T α 1Complete sequence or contain T α 1(17-24) any fragment of sequence can be synthetic with conventional solid phase or liquid phase polypeptide synthesis, in building-up process, be easy to introduce a Cys at N-end or C-end, the peptide chain that will contain Cys is soluble in water, transfer pH7-8 with sodium bicarbonate, add the reaction of 3 times of normal PEG-MAL or PEG-MAL or PEG-VS and PEG-IODO stirring at room, get the T α that PEG modifies with the RPLC purifying 1Or contain T α 1(17-24) fragment of sequence.The reaction of PEG-MAL modification Cys-T is as follows:
The present invention is 750,2000 and 5000 CH with molecular-weight average 3O-PEG-OH (mPEG-OH) is a raw material, has synthesized the maleimide derivatives (mPEG-MAL) of Series P EG.Use the solid-phase polypeptide synthesis method,, polypeptide cracking from the resin is got off and get Cys-T and T-Cys-NH through RPLC (RP-HPLC) purifying respectively at the N-of T end and halfcystine molecule of C-end introducing 2With Cys-T or T-Cys-NH 2Soluble in water, transfer pH7-8 with sodium bicarbonate, add 3 times of normal mPEG-MAL, stirring at room reaction, RP-HPLC monitoring reaction process.After reaction was finished, having obtained molecular-weight average with the RP-HPLC purified product was that 750,2000 and 5000 mPEG-MAL modifies the N-end of T and the product that C-holds.Each compound is analyzed to simple spike through RP-HPLC, shows that through mass spectrum and amino acid composition analysis structure is correct.
The invention further relates to formula (II) compound:
[PEG-X-(CH 2)mCO-Y]z-T???????(II)
Wherein T represents natural T α 1Complete sequence or contain T α 1(17-24) any fragment of sequence, X=O, NH or NHCO, Y=O or NH, m=0-6, z=1-11, and the amino or the hydroxyl in site can be through one or more PEG molecule covalent modifications, as natural T α arbitrarily in the T sequence 1Amino and 14,17,19,20 lysine side-chain amino in the complete sequence behind the N-end deacetylation, 1,8,9, the pendant hydroxyl group of position Serine and the pendant hydroxyl group of 7,12,13 Threonines.
The invention further relates to formula (III) compound,
T-[CO-Y-PEG]z??????????????(III)
Wherein, T represents natural T α 1Complete sequence or contain T α 1(17-24) any fragment of sequence, Y=O or NH, z=1-10, and the carboxyl in site can be through one or more PEG molecule covalent modifications, as natural T α arbitrarily in the T sequence 1C-end carboxyl in the complete sequence, the side chain carboxyl group of the side chain carboxyl group of 2,6,15 aspartic acids and 10,18,21,24,25,27 L-glutamic acid.
According to the present invention, with PEG-OH or PEG-NH 2Earlier respectively with phosgene, oxalyl chloride, C 2-C 8Halogenated acid, C 3-C 8Dicarboxylic anhydride reaction, again with HOSu react PEG-X-(CH 2) mCOOSu (m=0-6).With solid phase or the synthetic T of liquid phase polypeptide synthesis.T is soluble in water, transfer pH7-8 with sodium bicarbonate, add an amount of PEG-X-(CH 2) mCO-OSu, the stirring at room reaction can realize the hydroxyl modified of PEG to N-end amino, Lys side chain amino, Ser or the Thr side chain radical of polypeptide.
According to the present invention, introduce the amino acid of amino, carboxyl or the modification of preparation PEGization earlier (as Fmoc-Lys (NH-COCH at the end of PEG 2-PEG)-OH, Fmoc-Glu (CO-NH-PEG)-OHFmoc-Asp (CO-NH-PEG)-OH), be coupled in the peptide sequence with liquid phase or solid phase method again and go, can realize the modification that the N-end is amino, C-holds carboxyl, Lys side chain amino, Asp or Glu side chain carboxyl group polypeptide.
According to the present invention, pharmaceutical composition of the present invention is made the various formulations that are applicable to that Mammals is used, and for example makes injection with N.F,USP MANNITOL as vehicle.
According to the present invention, used term " contains T α among the present invention 1(17-24) any fragment of sequence " be meant T α 117-24 aminoacid sequence and the increase and decrease at 17-24 amino acids sequence two ends, and sequence below preferred
R 1-Lys-Glu-Lys-Lys-Glu-Val-Val-Glu-R 2
R 1＝H、
Leu、
Asp-Leu、
Lys-Asp-Leu、
Thr-Lys-Asp-Leu、
Thr-Thr-Lys-Asp-Leu、
Ile-Thr-Thr-Lys-Asp-Leu、
Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Ala-Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Asp-Ala-Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Ser-Asp-Ala-Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu。
R 2＝H、Glu、Glu-Ala、Glu-Ala-Glu、Glu-Ala-Glu-Asn。
According to the present invention, have for example with immune deficiency or immunologic hypofunction diseases associated: hepatitis B, hepatitis C, nonsmall-cell lung cancer, melanoma or acquired immune deficiency syndrome (AIDS), the SARS that coronavirus causes.
Embodiment
Embodiment
Following embodiment represents illustrative embodiment of the present invention, but the present invention is not subjected to the restriction of these embodiment.The used molecular-weight average of embodiment is that 5000 mPEG-OH is the Sigma product, and solid-phase synthesized carrier Wang resin is the ACT product, and TFA is the ACROS product, and Rink amide resins, DCC, HOBT, HBTU, Fmoc-protection amino acid are Shanghai gill biochemical products.
Embodiment 1 Cys (mPEG 5000-MAL)-preparation of T α 1
1.1 mPEG 5000-MAL's is synthetic
Weighing m PEG 5000-OH 20g (4mmol) places the 250ml reaction flask, adds 100mlCH 2Cl 2, add 3.0ml Et again after the solid dissolving 3N (20mmmol) and 3.8g Ts-Cl (20mmol), the stirring at room reaction.After the TLC monitoring reaction was complete, rotary evaporation removed and desolvates, and added the 100ml anhydrous diethyl ether and was settled out solid, got 15.2g mPEG 5000-OTs, yield 70%.
With 15g mPEG 5000-OTs (3mmol) is dissolved in 30ml DMF, adds 1.68g (18mmol) potassium phthalimide, and 120 ℃ were reacted 4 hours.The pressure reducing and steaming solvent is dissolved in the 50ml dehydrated alcohol with resistates, adds the 2.0ml hydrazine hydrate, back flow reaction 4 hours.Rotary evaporation removes and desolvates, and resistates is dissolved in 30ml CH 2Cl 2, the elimination insolubles removes the filtrate rotary evaporation and desolvates, and is settled out solid with anhydrous diethyl ether, gets 12.5g mPEG 5000-NH 2, yield 83%.
With 1.25g mPEG 5000-NH 2Be dissolved in the 20ml dioxane, add maleic anhydride 0.1g, 80 ℃ of stirring reaction 30min.The pressure reducing and steaming solvent adds the 50ml anhydrous diethyl ether, and cooling is settled out solid, and filter collection solid gets 1.10g after the drying.The gained solid is dissolved in the 15ml diacetyl oxide, adds the 0.5g sodium acetate, 100 ℃ of stirring reaction 45min.The pressure reducing and steaming solvent dissolves resistates with methylene dichloride, filtrate is concentrated into dried, adds anhydrous diethyl ether, is settled out solid, after filter collection, the drying light yellow solid 0.61g mPEG 5000-MAL, yield 48%.
1.2 Cys-T α 1Synthetic
With the synthetic Cys-T α of solid phase method 1: with 0.5mmol Wang resin is solid phase carrier, and Fmoc-AA-OH (2.0mmol) is as raw material, and DCC (2.0mmol)-HOBT (2.0mmol) makes condensing agent by T α 1The synthetic full guard peptide chain of aminoacid sequence, 2.9g Fmoc-T α 1-resin.Take by weighing 290mgFmoc-T α 1-resin (0.1mmol), remove Fmoc with 20% piperidines/DMF after, with DCC (2.0mmol)-HOBT (2.0mmol) Fmoc-Cys (Trt)-OH (2.0mmol) is coupled on the resin, again with 20% piperidines/DMF remove behind the Fmoc Cys-T α 1-resin.Behind resin drying, make lysate with EDT-meta-cresol-TFA, 0 ℃ was reacted 90 minutes, and the filtering resin is removed TFA with the filtrate rotary evaporation, and add anhydrous diethyl ether and be settled out white solid, filter collection solid, soluble in water, frost drying gets white dry powder 120mg.The RP-HPLC purifying, FAB-MS analyzes, M+1 peak: 3170 (theoretical values: 3169).
1.3 Cys (mPEG 5000-MAL)-T α 1Synthetic
Will be through the Cys-T α behind the RP-HPLC purifying 1Soluble in water, transfer pH to 7-8 with sodium bicarbonate, add 3 times of normal mPEG 5000-MAL, room temperature reaction is with RP-HPLC monitoring reaction process and separated product.
Cys (mPEG 5000-MAL)-T α 1Analyze through MALDI-TOF-MS, a series of peaks are arranged near 8350, it is about 44 that adjacent two peak molecular weight differ, and has the typical structure feature of polyoxyethylene glycol.The composition of amino acid analysis of acid hydrolysis (6N aqueous hydrochloric acid, 110 ℃, 22 hours) conforms to theoretical value: Asp, 4.00 (4); Thr, 2.81 (3); Ser, 2.75 (3); Glu, 6.57 (6); Ala, 3.00 (3); Val, 2.58 (3); Ile, 1.00 (1); Leu, 1.08 (1); Lys, 4.31 (4).
Embodiment 2 T α 1(17-24)-Cys (mPEG 5000-MAL)-NH 2Preparation
2.1 T α 1(17-24)-Cys-NH 2Synthetic
Make carrier with 0.1mmol Rink amide resins, Fmoc-AA-OH (0.2mmol) is as raw material, and HBTU (0.2mmol)-NMM (0.3mmol) makes condensing agent, presses T α 1Aminoacid sequence (17-24) synthesizes the full guard peptide resin.Make lysate with EDT-meta-cresol-TFA, 0 ℃ was reacted 90 minutes, and the filtering resin is removed TFA with the filtrate rotary evaporation, and add anhydrous diethyl ether and be settled out white solid, filter collection solid, soluble in water, frost drying gets white dry powder 20mg.The RP-HPLC purifying, FAB-MS analyzes, M+1 peak: 1091 (theoretical values: 1090).
2.2 T α 1(17-24)-Cys (mPEG 5000-MAL)-NH 2Synthetic
Will be through the T of RP-HPLC purifying α 1(17-24)-Cys-NH 2Soluble in water, transfer pH to 7-8 with sodium bicarbonate, add 3 times of normal mPEG 5000-MAL, room temperature reaction 2 hours is used the RP-HPLC separated product.
T α 1(17-24)-Cys (mPEG 5000-MAL)-NH 2-T α 1Analyze through MALDI-TOF-MS, a series of peaks are arranged near 6182, it is about 44 that adjacent two peak molecular weight differ, and has the typical structure feature of polyoxyethylene glycol.The composition of amino acid analysis of acid hydrolysis (6N aqueous hydrochloric acid, 110 ℃, 22 hours) conforms to theoretical value: Glu, 3.41 (3); Val, 1.52 (2); Lys, 3.07 (3).
Embodiment 3 T α 1And analogue produces the effect that induces of IFN-γ to mouse boosting cell
Broken end sacrificed by exsanguination mouse, aseptic condition are taken out spleen down rapidly, put into RPMI1640 liquid.Be placed on spleen on the 100 order stainless (steel) wires or grind with nook closing member or triangle glass rod in the nylon wire, make splenocyte suspension.With medium centrifugal washing splenocyte once after, add Tris-NH 4Cl damping fluid (0.16M NH 4Cl and 0.17M Tris mixed by 9: 1, and pH7.2) effect is 3-5 minute, and dissolving RBC uses the medium centrifugal washed twice again.Expect blue dyeing with tongue, viable count needs more than 95%.With 10% calf serum, 1640 liquid cell concn is adjusted to 1.25 * 10 7Individual/mL is standby.Splenocyte is joined in aseptic 48 well culture plates of cultivation every hole 800 μ L; Add different concns testing sample shown in the table 1 and ConA 100 μ L (final concentration 0.5 μ g/mL and 1.0 μ g/mL) again, put 5%CO 2In the incubator, cultivated 24 hours for 37 ℃, the centrifuging and taking supernatant is with the content of mouse IFN-γ detection kit mensuration IFN-γ.
Table 1
The sample title Sample concentration (μ g/mL) ????????????IFN-γ(ng/mL)
????ConA ????(0.5μg/mL) ????ConA ????(1.0μg/mL)
RPMI-1640 ????- ????27.3 ????45.2
??Zadaxin ????0.1 ????1.0 ????10.0 ????37.8 ????33.9 ????25.8 ????9.5 ????22.8 ????16.8
??Tα 1 ????0.1 ????1.0 ????10.0 ????25.0 ????34.0 ????28.4 ????15.1 ????33.9 ????35.4
??Cys(mPEG 5000-MAL)-Tα 1 ????0.1 ????1.0 ????10.0 ????29.0 ????26.7 ????33.5 ????21.7 ????13.6 ????27.3
??Cys(mPEG 2000-MAL)-Tα 1 ????0.1 ????1.0 ????10.0 ????22.9 ????12.8 ????12.0 ????38.9 ????26.4 ????19.2
Annotate: Zadaxin is the commercialization T α of Sai Sheng company 1, the IFN-γ concentration of blank cell culture fluid supernatant is 13.8ng/mL.
The external anti-SARS virus experiment of embodiment 4 T α 1 derivative
The Vero E6 cell that the 96 hole plastic plates in blocks of growing are cultivated, incline nutrient solution after, the SARS virus that adds 100TCID50, adsorbed 2 hours, inclining virus, adds testing sample, final concentration is 100 μ g/ml, continue to cultivate 7 days, examine under a microscope Vero E6 cytopathy degree every day, the result is as shown in table 2:
Table 2
Group Concentration (μ g/ml) The result of 100TCID50
Virus control (not adding T α 1 derivative) ????0 ????4+/4
Cys(mPEG5000-MAL)-Tα1 ????100 ????4-/4
Cys(mPEG5000-MAL)-Tα1(17-24) ????100 ????4-/4
Tα1-Cys(mPEG5000-MAL)-NH2 ????100 ????4+/4
Annotate: 4-/4 all virus-free seedlings of hole of 4 expressions, promptly negative; 4+/4 holes of 4 expressions are all positive.
Conclusion: Cys (mPEG5000-MAL)-T α 1 and Cys (mPEG5000-MAL)-T α 1 (17-24) has external anti-SARS virus activity when 100 μ g/ml, T α 1-Cys (mPEG5000-MAL)-NH2 does not have external anti-SARS virus activity when 100 μ g/ml.

Claims (7)

1. formula (I) compound or derivatives thereof:
PEG-M-Cys-T???????(I)
Figure A031314980002C1
Or
Figure A031314980002C2
Or
Wherein PEG is RO (CH 2CH 2O) n-CH 2CH 2, R=H or CH 3, n=5-1000,
Cys is a halfcystine, and it links to each other with M group covalency by its side chain thioether atom; Cys also links to each other or links to each other with the C-end carboxyl formation amido linkage of T with its amino with the amino formation of the N-end amido linkage of its carboxyl and T, and the Cys position comprises between the two ends of T sequence, any two adjacent amino acid and the amino acid of alternative optional position,
T represents natural T α 1Complete sequence or contain T α 1(17-24) any fragment of sequence.
2. according to the compound or derivatives thereof of claim 1, wherein contain T α 1(17-24) any fragment of sequence is represented by following formula:
R 1-Lys-Glu-Lys-Lys-Glu-Val-Val-Glu-R 2
R 1＝H、
Leu、
Asp-Leu、
Lys-Asp-Leu、
Thr-Lys-Asp-Leu、
Thr-Thr-Lys-Asp-Leu、
Ile-Thr-Thr-Lys-Asp-Leu、
Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Ala-Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Asp-Ala-Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu、
Ser-Asp-Ala-Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu。
R 2＝H、Glu、Glu-Ala、Glu-Ala-Glu、Glu-Ala-Glu-Asn。
3. formula (II) compound:
[PEG-X-(CH 2)mCO-Y]z-T???????????(II)
Wherein T represents natural T α 1Complete sequence or contain T α 1(17-24) any fragment of sequence, X=O, NH or NHCO, Y=O or NH, m=0-6, z=1-11, and the amino or the hydroxyl in site can be through one or more PEG molecule covalent modifications, as natural T α arbitrarily in the T sequence 1Amino and 14,17,19,20 lysine side-chain amino in the complete sequence behind the N-end deacetylation, 1,8,9, the pendant hydroxyl group of position Serine and the pendant hydroxyl group of 7,12,13 Threonines.
4. formula (III) compound,
T-[CO-Y-PEG] z (III) wherein, T represents natural T α 1Complete sequence or contain T α 1(17-24) any fragment of sequence, Y=O or NH, z-1-10, and the carboxyl in site can be through one or more PEG molecule covalent modifications, as natural T α arbitrarily in the T sequence 1C-end carboxyl in the complete sequence, the side chain carboxyl group of the side chain carboxyl group of 2,6,15 aspartic acids and 10,18,21,24,25,27 L-glutamic acid.
5. pharmaceutical composition, it contains compound and the pharmaceutical carrier or the vehicle of at least a formula (I) or formula (II) or formula (III).
6. arbitrary compound or derivative are used for preventing or the medicine purposes of treatment and immune deficiency or immunologic hypofunction relative disease is included in hepatitis B in preparation among the claim 1-4, hepatitis C, malignant melanoma, application in the treatment of the relative diseases such as SARS that lung cancer in non-cellule type, coronavirus cause or the prevention.
7. the method for preparation formula (I) compound or derivative, it comprises PEG-MAL or PEG-VS or PEG-IODO and HS-Cys-T reaction.
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Cited By (10)

* Cited by examiner, † Cited by third party
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WO2005105151A1 (en) * 2004-04-28 2005-11-10 Jiangsu Hansen Pharmaceutical Co., Ltd. Pegylated thymosin 1 derivatives
WO2007054030A1 (en) * 2005-11-10 2007-05-18 Institute Of Pharmacology And Toxicology Academy Of Military Medical Sciences P.L.A. China Polyethylene glycol modifications of thymosin alpha-1
CN100381420C (en) * 2005-03-23 2008-04-16 于勇海 N-acetyl cysteine derivatives and use thereof
WO2008151512A1 (en) * 2007-06-12 2008-12-18 Institute Of Pharmacology And Toxicology Academy Of Military Medical Sciences P.L.A. Site-specific pegylated linear salmon calcitonin derivatives
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Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4179337A (en) * 1973-07-20 1979-12-18 Davis Frank F Non-immunogenic polypeptides
US5308833A (en) * 1992-02-06 1994-05-03 Sciclone Pharmaceuticals Method for treating hepatitis B carriers with minimal disease

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