CN101899085A - Saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester, synthetic method and application thereof - Google Patents

Saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester, synthetic method and application thereof Download PDF

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CN101899085A
CN101899085A CN2009100853198A CN200910085319A CN101899085A CN 101899085 A CN101899085 A CN 101899085A CN 2009100853198 A CN2009100853198 A CN 2009100853198A CN 200910085319 A CN200910085319 A CN 200910085319A CN 101899085 A CN101899085 A CN 101899085A
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asp
saturated fatty
gly
glu
obzl
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赵明
彭师奇
赵淑锐
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Capital Medical University
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Capital Medical University
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Abstract

The invention relates to 6 saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester conjugates with immunosuppressive activity in a general formula Glu-Asp-Gly-O-CH2-(CH2)nCH3 I. In saturated fatty chain alcohol, n is equal to 6, 8, 10, 12, 14 or 16. The invention also relates to a preparation method and application of the conjugates as an immunosuppressant. An experimental result of the saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester has the inhibition effects of splenic lymphocyte mitogen breeder reaction and macrophage phagocytosis activity indicates that the conjugates of the invention have excellent immunosuppressive action and can be clinically used as an immunosuppressant.

Description

Saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester and synthetic method thereof and application
Technical field
The present invention relates to the saturated aliphatic chain alcohol tripeptide ester, relate in particular to a series of saturated fatty chain alcohol Glu-Asp-Gly tripeptide esters with immunosuppressive activity, also relate to the preparation method of these saturated aliphatic chain alcohol tripeptide esters and they as the application of immunosuppressor, belong to biomedicine field.
Background technology
According to statistics, by the end of the year 2002, various organ transplantations 935792 examples time are carried out in the whole world altogether, wherein renal transplantation 585877 examples time, liver transplantation 112153 examples time, heart transplantation 66559 examples time.In addition, many internal organs combined transplantation such as organ transplantation such as pancreas, lung, small intestine and the heart-lung, pancreas-kidney, liver-kidney, liver-intestines is also all succeedd and is applied to clinical.At present, 1 year people/kidney survival rate of renal transplantation reaches 90%～95%, and 5 annual survival rates surpass 70%.Organ transplantation will form transplantation tolerance, and this just requires the patient to take immunosuppressor all the life.The progress of organ transplantation depends on the progress of immunosuppressor to a great extent.Nearly decades, though new immunosuppressive drug makes organ transplantation get the development of advancing by leaps and bounds clinical, their toxic side effect, for example renal toxicity and bone marrow inhibition remain organ transplantation and make the serious problems that must face.
Cyclosporin A is present clinical immunosuppressor commonly used.Because the water-soluble extreme difference and the renal toxicity of cyclosporin A are very strong, so be that formulation or curative effect are all unsatisfactory.Improve the water-soluble of cyclosporin A preparation, reduce the renal toxicity of cyclosporin A, be the focus of cyclosporin A research always.The urotoxin peptide has immunosuppressive activity.The contriver recognizes that the ester that help a small child urinate by holding his legs apart toxin peptide and fatty chain alcohol are puted together generation has the self-assembly performance, thereby can be as the medicine carrying material with immunosuppressive activity.For example can be used for wrapping up cyclosporin A, reach the dual purpose that improves the water-soluble of cyclosporin A and reduce the cyclosporin A renal toxicity.
Summary of the invention
One of purpose of the present invention is, urotoxin tripeptides and fatty chain alcohol are puted together, and obtains having the saturated aliphatic chain alcohol tripeptide ester conjugate of immunosuppressive activity.
One of purpose of the present invention is achieved through the following technical solutions:
6 kinds of saturated fatty chain alcohol Glu-Asp-Gly tripeptide esters of general formula I with immunosuppressive activity
Glu-Asp-Gly-O-CH 2-(CH 2)nCH 3 I
Wherein, the n=6 in the general formula I, 8,10,12,14 or 16.
Two of purpose of the present invention is that a kind of above-mentioned method with saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester of immunosuppressive activity for preparing is provided.
Two of purpose of the present invention is achieved through the following technical solutions:
A kind of 6 kinds of methods that prepare general formula I with saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester of immunosuppressive activity, this method comprises:
(1) glycine and the saturated fatty alcohol condensation of N being held protecting group protect, described saturated fatty alcohol is CH 3(CH 2) nCH 2OH, n=6,8,10,12,14 or 16;
(2) slough N end protecting group and obtain the saturated fatty alcohol glycinate;
(3), the protection intermediate of saturated fatty alcohol glycinate and aspartic acid, the protection intermediate of L-glutamic acid are progressively connect saturated fatty alcohol three peptide conjugates of the synthetic protecting group protection of peptide according to existing liquid phase synthetic technology;
(4) N end protecting group and the C that sloughs saturated fatty alcohol three peptide conjugates of protecting group protection successively holds protecting group to obtain target compound.
Wherein said N end protecting group is a blocking group commonly used when the N end of polypeptide is protected, and for example can be tertbutyloxycarbonyl (Boc); Described C end protecting group is a blocking group commonly used when the C end of polypeptide is protected, and for example can be benzyloxy (OBzl); The process of described liquid phase synthetic technology and described protection, condensation, deprotection is the conventional and technique known of this area.
This preparation method can summarize with the route of Fig. 1, and concrete, described method comprises:
(1) at (Boc) 2O and NaOH are converted into the N-t-butoxycarbonyl glycine with glycine under existing;
(2) in the presence of DCC, HOBt, anhydrous THF with N-t-butoxycarbonyl glycine and saturated fatty alcohol condensation, generate saturated fatty alcohol N-t-butoxycarbonyl glycine ester;
(3) in hydrogenchloride-ethyl acetate, saturated fatty alcohol N-t-butoxycarbonyl glycine ester is removed the tertiary butyloxycarbonyl protecting group, generate the saturated fatty alcohol glycinate;
(4) in anhydrous THF, in the presence of DCC and HOBt,, generate saturated fatty alcohol N-tertbutyloxycarbonyl-β-benzyl ester aspartoyl glycinate with saturated fatty alcohol glycinate and N-tertbutyloxycarbonyl-β-benzyl ester aspartic acid condensation;
(5) in the presence of hydrogenchloride-ethyl acetate, saturated fatty alcohol N-tertbutyloxycarbonyl-β-benzyl ester aspartoyl glycinate is removed tertbutyloxycarbonyl and generate saturated fatty alcohol β-benzyl ester aspartoyl glycinate;
(6) in anhydrous THF, in the presence of DCC and HOBt,, generate saturated fatty alcohol N-tertbutyloxycarbonyl-γ-benzyl ester glutamy β-benzyl ester aspartoyl glycinate with saturated fatty alcohol β-benzyl ester-aspartoyl glycinate and N-tertbutyloxycarbonyl-γ-benzyl ester-L-glutamic acid condensation;
(7) in hydrogenchloride-ethyl acetate, saturated fatty alcohol N-tertbutyloxycarbonyl-γ-benzyl ester glutamy β-benzyl ester aspartoyl glycinate is removed tertbutyloxycarbonyl, generate saturated fatty alcohol γ-benzyl ester glutamy β-benzyl ester aspartoyl glycinate;
(8) in dehydrated alcohol, in the presence of Pd/C,, generate saturated fatty alcohol glutamy aspartoyl glycinate with saturated fatty alcohol γ-benzyl ester glutamy β-benzyl ester aspartoyl glycinate hydrogenolysis.
Experimental result shows that 6 kinds of saturated fatty chain alcohol Glu-Asp-Gly tripeptide esters with immunosuppressive activity of the present invention have outstanding immunosuppressive action, can be used as immunosuppressor clinically and uses.And equal energy self-assembly granulating is through being stabilized in the nanometer ball of 117-2290nm in the saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester aqueous solution of the present invention, can be used as the preparation material of preparation micro emulsion or lipidosome drug carrier, can be used as the targeting preparation material of preparation micro emulsion, lipidosome drug carrier in addition.
Description of drawings
Fig. 1 is 6 kinds of synthetic routes with saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester of immunosuppressive activity of general formula I.I) anhydrous THF, DCC, HOBt and NMM; Ii) 4N hydrogenchloride-ethyl acetate solution; Iii) dehydrated alcohol, Pd/C and H 27a n=6,7b n=8,7c n=10,7d n=12,7e n=14,7f n=16.
In order further to set forth the present invention, provide a series of embodiment below.These embodiment are illustrative fully, and they only are used for the present invention is specifically described, and not should be understood to limitation of the present invention.
Embodiment
Embodiment 1Boc-Gly-OCH 2(CH 2) 6CH 3Preparation
1.00g (5.71mmol) Boc-Gly is dissolved in the anhydrous THF of 20ml, and ice bath adds 0.77g (5.71mmol) N-hydroxy benzo triazole (HOBt) down in the solution that obtains, and makes dissolving fully.Add 1.41g (6.85mmol) dicyclohexyl carbonyl diimine (DCC) after 10 minutes, obtain reaction solution (I).The following 0.74g of ice bath (5.71mmol) Fatty Alcohol(C12-C14 and C12-C18) CH 3(CH 2) 6CH 2OH is suspended among the anhydrous THF of 20ml, adds 1ml N-methylmorpholine (NMM) then, transfers pH 8-9.Stirred 35 minutes, and obtained reaction solution (II).The following reaction solution of ice bath (I) adds in the reaction solution (II), and first ice bath stirs 1h down, stirring at room 12h again, and TLC (chloroform/methanol, 10: 1) shows that Boc-Lys (Z) disappears.Filtering dicyclohexylurea (DCU) (DCU), THF is removed in decompression.Residue 50ml acetic acid ethyl dissolution.The solution that obtains is used saturated NaHCO successively 3The aqueous solution is washed, the saturated NaCl aqueous solution is washed, 5%KHSO 4The aqueous solution is washed with the saturated NaCl aqueous solution and is washed.The organic phase anhydrous Na 2SO 4Drying, filtration, filtrate decompression are concentrated into dried, obtain 1.57g (96%) title compound, are colorless oil.ESI-MS(m/z)：288[M+H] +。
Embodiment 2Boc-Gly-OCH 2(CH 2) 8CH 3Preparation
According to the method for embodiment 1 by 1.00g (5.71mmol) Boc-Gly and 0.90g (5.71mmol) CH 3(CH 2) 8CH 2OH makes 1.64g (91%) title compound, is colorless oil ESI-MS (m/z): 316[M+H] +
Embodiment 3Boc-Gly-OCH 2(CH 2) 10CH 3Preparation
According to the method for embodiment 1 by 1.00g (5.71mmol) Boc-Gly and 1.06g (5.71mmol) CH 3(CH 2) 10CH 2OH makes 1.77g (90%) title compound, is colorless oil.ESI-MS(m/z)：344[M+H] +。
Embodiment 4Boc-Gly-OCH 2(CH 2) 12CH 3Preparation
According to the method for embodiment 1 by 1.00g (5.71mmol) Boc-Gly and 1.22g (5.71mmol) CH 3(CH 2) 12CH 2OH makes 1.88g (89%) title compound, is colorless oil.ESI-MS(m/z)：372[M+H] +。
Embodiment 5Boc-Gly-OCH 2(CH 2) 14CH 3Preparation
According to the method for embodiment 1 by 1.00g (5.71mmol) Boc-Gly and 1.38g (5.71mmol) CH 3(CH 2) 14CH 2OH makes 1.96g (86%) title compound, is colorless oil.ESI-MS(m/z)：400[M+H] +。
Embodiment 6Boc-Gly-OCH 2(CH 2) 16CH 3Preparation
According to the method for embodiment 1 by 1.00g (5.71mmol) Boc-Gly and 1.54g (5.71mmol) CH 3(CH 2) 16CH 2OH makes 1.97g (81%) title compound, is colorless oil.ESI-MS(m/z)：428[M+H] +。
Embodiment 7HClGly-OCH 2(CH 2) 6CH 3Preparation
With 1.57g (5.47mmol) Boc-Gly-OCH 2(CH 2) 6CH 3Be dissolved in 50ml 4N hydrogenchloride-ethyl acetate solution, stirring at room 2 hours, TLC (chloroform: methyl alcohol, 5: 1) shows Boc-Gly-OCH 2(CH 2) 6CH 3Disappear, concentrating under reduced pressure is removed ethyl acetate, and residue adds a small amount of ether repeatedly and carries out concentrating under reduced pressure to remove de-chlorine hydride.Add a small amount of ether at last residue is ground to form 1.14g (93%) title compound, be beige oily matter.ESI-MS(m/z)：188[M+H] +。
Embodiment 8HClGly-OCH 2(CH 2) 8CH 3Preparation
According to the method for embodiment 7, from 1.64g (5.21mmol) Boc-Gly-OCH 2(CH 2) 8CH 3Make 1.23g (94%) title compound, be beige oily matter.ESI-MS(m/z)：216[M+H] +。
Embodiment 9HClGly-OCH 2(CH 2) 10CH 3Preparation
According to the method for embodiment 7, from 1.77g (5.16mmol) Boc-Gly-OCH 2(CH 2) 10CH 3Make 1.30g (90%) title compound, be beige oily matter.ESI-MS(m/z)：244[M+H] +。
Embodiment 10HClGly-OCH 2(CH 2) 12CH 3Preparation
According to the method for embodiment 7, from 1.88g (5.07mmol) Boc-Gly-OCH 2(CH 2) 12CH 3Make 1.37g (88%) title compound, be beige oily matter.ESI-MS(m/z)：272[M+H] +。
Embodiment 11HClGly-OCH 2(CH 2) 14CH 3Preparation
According to the method for embodiment 7, from 1.96g (4.91mmol) Boc-Gly-OCH 2(CH 2) 14CH 3Make 1.49g (91%) title compound, be beige oily matter.ESI-MS(m/z)：300[M+H] +。
Embodiment 12HClGly-OCH 2(CH 2) 16CH 3Preparation
According to the method for embodiment 7, from 1.97g (4.61mmol) Boc-Gly-OCH 2(CH 2) 16CH 3Make 1.59g (95%) title compound, be beige oily matter.ESI-MS(m/z)：328[M+H] +。
Embodiment 13Boc-Asp (OBzl)-Gly-OCH 2(CH 2) 6CH 3Preparation
According to the method for embodiment 1 by 1.65g (5.10mmol) Boc-Asp (OBzl) and 1.14g (5.10mmol) HClGly-OCH 2(CH 2) 6CH 3Make 2.03g (81%) title compound, be faint yellow oily thing.ESI-MS(m/z)：493[M+H] +，[α] 20 D＝-11.0(c＝1.0，CH 3OH)。
Embodiment 14Boc-Asp (OBzl)-Gly-OCH 2(CH 2) 8CH 3Preparation
According to the method for embodiment 1 by 1.58g (4.89mmol) Boc-Asp (OBzl) and 1.23g (4.89mmol) HClGly-OCH 2(CH 2) 8CH 3Make 2.03g (80%) title compound, be faint yellow oily thing.ESI-MS(m/z)：521[M+H] +，[α] 20 D＝-10.0(c＝1.0，CH 3OH)。
Embodiment 15Boc-Asp (OBzl)-Gly-OCH 2(CH 2) 10CH 3Preparation
According to the method for embodiment 1 by 1.50g (4.65mmol) Boc-Asp (OBzl) and 1.30g (4.65mmol) HClGly-OCH 2(CH 2) 10CH 3Make 2.06g (81%) title compound, be colorless oil.ESI-MS(m/z)：549[M+H] +，[α] 20 D＝-5.0(c＝1.0，CH 3OH)。
Embodiment 16Boc-Asp (OBzl)-Gly-OCH 2(CH 2) 12CH 3Preparation
According to the method for embodiment 1 by 1.44g (4.46mmol) Boc-Asp (OBzl) and 1.37g (4.46mmol) HClGly-OCH 2(CH 2) 12CH 3Make 2.18g (85%) title compound, be faint yellow oily thing.ESI-MS(m/z)：577[M+H] +，[α] 20 D＝-5.8(c＝1.0，CH 3OH)。
Embodiment 17Boc-Asp (OBzl)-Gly-OCH 2(CH 2) 14CH 3Preparation
According to the method for embodiment 1 by 1.43g (4.44mmol) Boc-Asp (OBzl) and 1.49g (4.44mmol) HClGly-OCH 2(CH 2) 14CH 3Make 2.20g (82%) title compound, be faint yellow solid.ESI-MS(m/z)：605[M+H] +，[α] 20 D＝-4.5(c＝1.0，CH 3OH)。
Embodiment 18Boc-Asp (OBzl)-Gly-OCH 2(CH 2) 16CH 3Preparation
According to the method for embodiment 1 by 1.41g (4.37mmol) Boc-Asp (OBzl) and 1.59g (4.37mmol) HClGly-OCH 2(CH 2) 16CH 3Make 2.19g (79%) title compound, be faint yellow solid.ESI-MS(m/z)：633[M+H] +，[α] 20 D＝-4.2(c＝1.0，CH 3OH)。
Embodiment 19HClAsp (OBzl)-Gly-OCH 2(CH 2) 6CH 3Preparation
According to the method for embodiment 7, from 2.03g (4.12mmol) Boc-Asp (OBzl)-Gly-OCH 2(CH 2) 6CH 3Make 1.61g (91%) title compound, be beige oily matter.ESI-MS(m/z)：393[M+H] +，[α] 20 D＝-5.7(c＝1.0，CH 3OH)。
Embodiment 20HClAsp (OBzl)-Gly-OCH 2(CH 2) 8CH 3Preparation
According to the method for embodiment 7, from 2.03g (3.90mmol) Boc-Asp (OBzl)-Gly-OCH 2(CH 2) 8CH 3Make 1.64g (92%) title compound, be beige oily matter.ESI-MS(m/z)：421[M+H] +，[α] 20 D＝-9.5(c＝1.0，CH 3OH)。
Embodiment 21HClAsp (OBzl)-Gly-OCH 2(CH 2) 10CH 3Preparation
According to the method for embodiment 7, from 2.06g (3.65mmol) Boc-Asp (OBzl)-Gly-OCH 2(CH 2) 10CH 3Make 1.69g (96%) title compound, be beige oily matter.ESI-MS(m/z)：449[M+H] +，[α] 20 D＝9.3(c＝1.0，CH 3OH)。
Embodiment 22HClAsp (OBzl)-Gly-OCH 2(CH 2) 12CH 3Preparation
According to the method for embodiment 7, from 2.18g (3.78mmol) Boc-Asp (OBzl)-Gly-OCH 2(CH 2) 12CH 3Make 1.76g (91%) title compound, be beige oily matter.ESI-MS(m/z)：477[M+H] +，[α] 20 D＝-10.5(c＝1.0，CH 3OH)。
Embodiment 23HClAsp (OBzl)-Gly-OCH 2(CH 2) 14CH 3Preparation
According to the method for embodiment 7, from 2.20g (3.64mmol) Boc-Asp (OBzl)-Gly-OCH 2(CH 2) 14CH 3Make 1.87g (95%) title compound, be the beige solid.ESI-MS(m/z)：505[M+H] +，[α] 20 D＝5.5(c＝1.0，CH 3OH)。
Embodiment 24HClAsp (OBzl)-Gly-OCH 2(CH 2) 16CH 3Preparation
According to the method for embodiment 7, from 2.19g (3.47mmol) Boc-Asp (OBzl)-Gly-OCH 2(CH 2) 16CH 3Make 1.89g (96%) title compound, be the beige solid.ESI-MS(m/z)：533[M+H] +，
[α] 20 D＝5.3(c＝1.0，CH 3OH)。
Embodiment 25Boc-Glu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 6CH 3Preparation
According to the method for embodiment 1 by 1.27g (3.76mmol) Boc-Glu (OBzl) and 1.61g (3.76mmol) HClAsp (OBzl)-Gly-OCH 2(CH 2) 6CH 3Make, the gained compound gets water white transparency oily thing 1.76g through purification by silica gel column chromatography, purification condition: sherwood oil: acetone=3: 1, yield are 66%.ESI-MS(m/z)：712[M+H] +，[α] 20 D＝-29.3(c＝1.0，CH 3OH)。
Embodiment 26Boc-Glu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 8CH 3Preparation
According to the method for embodiment 1 by 1.21g (3.59mmol) Boc-Glu (OBzl) and 1.64g (3.59mmol) HClAsp (OBzl)-Gly-OCH 2(CH 2) 8CH 3Make, the gained compound gets water white transparency oily thing 1.88g through purification by silica gel column chromatography, purification condition: sherwood oil: acetone=3: 1, yield are 71%.ESI-MS(m/z)：740[M+H] +，[α] 20 D＝-27.4(c＝1.0，CH 3OH)。
Embodiment 27Boc-Glu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 10CH 3Preparation
According to the method for embodiment 1 by 1.18g (3.49mmol) Boc-Glu (OBzl) and 1.69g (3.49mmol) HClAsp (OBzl)-Gly-OCH 2(CH 2) 10CH 3Make, the gained compound gets water white transparency oily thing 1.45g through purification by silica gel column chromatography, purification condition: sherwood oil: acetone=3: 1, yield are 54%.ESI-MS(m/z)：768[M+H] +，[α] 20 D＝-24.2(c＝1.0，CH 3OH)。
Embodiment 28Boc-Glu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 12CH 3Preparation
According to the method for embodiment 1 by 1.16g (3.43mmol) Boc-Glu (OBzl) and 1.76g (3.43mmol) HClAsp (OBzl)-Gly-OCH 2(CH 2) 12CH 3Make, the gained compound gets water white transparency oily thing 1.91g through purification by silica gel column chromatography, purification condition: sherwood oil: acetone=3: 1, yield are 70%.ESI-MS(m/z)：796[M+H] +，[α] 20 D＝-18.6(c＝1.0，CH 3OH)。
Embodiment 29Boc-Glu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 14CH 3Preparation
According to the method for embodiment 1 by 1.17g (3.46mmol) Boc-Glu (OBzl) and 1.87g (3.46mmol) HClAsp (OBzl)-Gly-OCH 2(CH 2) 14CH 3Make, the gained compound gets water white transparency oily thing 1.91g through purification by silica gel column chromatography, purification condition: sherwood oil: acetone=3: 1, yield are 67%.ESI-MS(m/z)：824[M+H] +，[α] 20 D＝-26.2(c＝1.0，CH 3OH)。
Embodiment 30Boc-Glu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 16CH 3Preparation
According to the method for embodiment 1 by 1.12g (3.32mmol) Boc-Glu (OBzl) and 1.89g (3.32mmol) HClAsp (OBzl)-Gly-OCH 2(CH 2) 16CH 3Make, the gained compound gets water white transparency oily thing 1.61g through purification by silica gel column chromatography, purification condition: sherwood oil: acetone=3: 1, yield are 57%.ESI-MS(m/z)：852[M+H] +，[α] 20 D＝-19.4(c＝1.0，CH 3OH)。
Embodiment 31HClGlu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 6CH 3Preparation
According to the method for embodiment 7, from 1.20g (1.69mmol) Boc-Glu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 6CH 3Make 0.98g (90%) title compound, be colorless solid.ESI-MS(m/z)：612[M+H] +。
Embodiment 32HClGlu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 8CH 3Preparation
According to the method for embodiment 7, from 1.30mg (1.76mmol) Boc-Glu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 8CH 3Make 1.08g (91%) title compound, be colorless solid.ESI-MS(m/z)：640[M+H] +。
Embodiment 33HClGlu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 10CH 3Preparation
According to the method for embodiment 7, from 1.20mg (1.56mmol) Boc-Glu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 10CH 3Make 1.01g (92%) title compound, be colorless solid.ESI-MS(m/z)：668[M+H] +。
Embodiment 34HClGlu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 12CH 3Preparation
According to the method for embodiment 7, from 1.50mg (1.89mmol) Boc-Glu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 12CH 3Make 1.30g (94%) title compound, be colorless solid.ESI-MS(m/z)：696[M+H] +。
Embodiment 35HClGlu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 14CH 3Preparation
According to the method for embodiment 7, from 1.50g (1.82mmol) Boc-Glu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 14CH 3Make 1.26g (91%) title compound, be colorless solid.ESI-MS(m/z)：724[M+H] +。
Embodiment 36HClGlu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 16CH 3Preparation
According to the method for embodiment 7, from 1.41g (1.66mmol) Boc-Glu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 16CH 3Make 1.17g (89%) title compound, be colorless solid.ESI-MS(m/z)：752[M+H] +。
Embodiment 37Glu-Asp-Gly-OCH 2(CH 2) 6CH 3Preparation (7a)
With 980mg (1.52mmol) HClGlu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 6CH 3With dissolve with ethanol, add an amount of Pd/C (about 5%), logical H 2(0.02Mba), stirring at room to raw material point disappears.Filtering Pd/C, filtrate decompression are concentrated into dried.Make 554mg (84%) title compound, be colorless oil.ESI-MS(m/z)：430[M-H] -，[α] 20 D＝-15.9(c＝1.0，CH 3OH)。
Embodiment 38Glu-Asp-Gly-OCH 2(CH 2) 8CH 3Preparation (7b)
With 1.08g (1.60mmol) HClGlu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 8CH 3With dissolve with ethanol, add an amount of Pd/C (about 5%), logical H 2(0.02Mba), stirring at room to raw material point disappears.Filtering Pd/C, filtrate decompression are concentrated into dried.Make 646mg (88%) title compound, be colorless oil.ESI-MS(m/z)：458[M-H] -，[α] 20 D＝-17.8(c＝1.0，CH 3OH)。
Embodiment 39Glu-Asp-Gly-OCH 2(CH 2) 10CH 3Preparation (7c)
With 1.00mg (1.44mmol) HClGlu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 10CH 3With dissolve with ethanol, add an amount of Pd/C (about 5%), logical H 2(0.02Mba), stirring at room to raw material point disappears.Filtering Pd/C, filtrate decompression are concentrated into dried.Make 632mg (90%) title compound, be colorless oil.ESI-MS(m/z)：486[M-H] -，[α] 20 D＝-17.0(c＝1.0，CH 3OH)。
Embodiment 40Glu-Asp-Gly-OCH 2(CH 2) 12CH 3Preparation (7d)
With 1.30g (1.78mmol) HClGlu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 12CH 3With dissolve with ethanol, add an amount of Pd/C (about 5%), logical H 2(0.02Mba), stirring at room to raw material point disappears.Filtering Pd/C, filtrate decompression are concentrated into dried.Make 806mg (88%) title compound, be colorless solid.ESI-MS(m/z)：514[M-H] -，[α] 20 D＝-6.8(c＝1.0，CH 3OH)，M.p.：88.0-89.0℃。
Embodiment 41Glu-Asp-Gly-OCH 2(CH 2) 14CH 3Preparation (7e)
With 1.26g (1.66mmol) HClGlu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 14CH 3With dissolve with ethanol, add an amount of Pd/C (about 5%), logical H 2(0.02Mba), stirring at room to raw material point disappears.Filtering Pd/C, filtrate decompression are concentrated into dried.Make 869mg (95%) title compound, be colorless solid.ESI-MS(m/z)：542[M-H] -；，[α] 20 D＝-11.1(c＝1.0，CH 3OH)，M.p.：78.8-79.2℃。
Embodiment 42Glu-Asp-Gly-OCH 2(CH 2) 16CH 3Preparation (7f)
With 1.17g (1.48mmol) HClGlu (OBzl)-Asp (OBzl)-Gly-OCH 2(CH 2) 16CH 3With dissolve with ethanol, add an amount of Pd/C (about 5%), logical H 2(0.02Mba), stirring at room to raw material point disappears.Filtering Pd/C, filtrate decompression are concentrated into dried.Make 823mg (97%) title compound, be colorless solid.ESI-MS(m/z)：570[M-H] -，[α] 20 D＝-5.5(c＝1.0，CH 3OH)，M.p.：80.8-82.2℃。
Test example 1 saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester is to the effect of mouse spleen lymphocyte mitogen inhibition of proliferation
Take off neck and put to death mouse, the aseptic spleen of getting grinds with 200 order steel meshes and piston, use HANK ' S liquid to wash under twice, 1500 rev/min of condition centrifugal 10 minutes, counts then being made into 5 * 10 with complete RPMI-1640 nutrient solution 6/ ml splenic lymphocyte, (every hole contains 5 * 10 in 96 well culture plates to add 100 μ l cell suspensions 6Individual cell).Every hole adds 20 μ l ConA (the ConA final concentration is 5 μ g/ml), and it is 0.05 CO that this 96 well culture plate places volume fraction 2Cultivate 4h for 37 ℃ in the incubator of saturated humidity.Add different concns saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester (1 * 10 behind the 4h respectively -4M, 8 * 10 -5M, 5 * 10 -5M, 3 * 10 -5M, 1 * 10 -5M, 8 * 10 -6M, 5 * 10 -6M and 1 * 10 -6M), 3 multiple holes of each concentration.Establish simultaneously and do not contain the compound control wells and only contain the cell blank hole of not having ConA with the amount nutrient solution.(n=3) all repeated in each hole 3 times.Use the restraining effect of mtt assay detection compound after cultivating 48h to splenic lymphocyte.
Calculate the restraining effect of different concns saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester according to formula " inhibiting rate=(D contrast-D pastille)/D contrast * 100% " to spleen lymphocyte proliferation, draw cell growth curve according to cell relative survival rate and compound concentrations, utilize this growth curve try to achieve the half inhibiting rate ( DxIC 50).The result lists table 1 in.The result shows that saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester of the present invention has clear and definite restraining effect to mice spleen lymphocytes proliferation.
Table 1 saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester is to the effect of mouse spleen lymphocyte mitogen inhibition of proliferation
Test example 2 saturated fatty chain alcohol Glu-Asp-Gly tripeptide esters are to the restraining effect of macrophage phagocytic
Growth conditions is good, be in the Ana-1 mouse macrophage of logarithmic phase with 1 * 10 5The density of individual/mL is inoculated in 96 orifice plates, every hole 100 μ l, 37 ℃, 5%CO 2Cultivated 4 hours in the incubator, by the saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester (1 * 10 of default concentration gradient adding through sterilising treatment -4M, 8 * 10 -5M, 5 * 10 -5M, 3 * 10 -5M, 1 * 10 -5M, 8 * 10 -6M, 5 * 10 -6M and 1 * 10 -6M), 3 multiple holes of each concentration, control group adds the solvent of isopyknic sample dissolution.Continue to cultivate after 24 hours, inhale and abandon supernatant liquor, every hole adds the neutral red solution of 100 μ l0.1%, places 37 ℃ to hatch 30 minutes.Neutral red solution is abandoned in suction, and washs 2-3 time (removing not by the toluylene red of macrophage phagocytic) with PBS buffered soln, and (ethanol: 100 μ l acetate=1: 1), 4 ℃ are spent the night, microplate reader detection absorbance, wavelength 540nm to add cytolysate.(n=3) all repeated in each hole 3 times.
Calculate the restraining effect of the compound of different concns according to formula " inhibiting rate=(D contrast-D pastille)/D contrast * 100% " to the phagocytic activity of scavenger cell, draw cell growth curve according to cell relative survival rate and compound concentrations, utilize this growth curve try to achieve the half inhibiting rate ( DxIC 50).The result lists table 2 in.The result shows that saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester of the present invention engulfs the Ana-1 mouse macrophage clear and definite restraining effect is arranged.
The restraining effect that table 2 saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester is engulfed the Ana-1 mouse macrophage
Figure B2009100853198D0000121
Test example 3 saturated fatty chain alcohol Glu-Asp-Gly tripeptide esters are in the self-assembly of nanometer level
Saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester is configured to the aqueous solution of 1 μ mol/ml, on the laser nano particle size analyzer, measures particle diameters for 25 ℃.METHOD FOR CONTINUOUS DETERMINATION 8 days writes down its particle diameter.The result lists table 3 in.Data show that equal energy self-assembly granulating is through being stabilized in the nanometer ball of 117-2290nm in the saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester aqueous solution of the present invention.
The nanometer ball particle diameter of table 3 saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester self-assembly in water

Claims (5)

1. 6 of general formula I kinds of saturated fatty chain alcohol Glu-Asp-Gly tripeptide esters with immunosuppressive activity:
Glu-Asp-Gly-O-CH 2-(CH 2)nCH 3 I
N=6 in the general formula I, 8,10,12,14 or 16.
2. method for preparing the saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester of claim 1 comprises:
(1) glycine and the saturated fatty alcohol condensation of N being held protecting group protect, described saturated fatty alcohol is CH 3(CH 2) nCH 2OH, n=6,8,10,12,14 or 16;
(2) slough N end protecting group and obtain the saturated fatty alcohol glycinate;
(3), the protection intermediate of saturated fatty alcohol glycinate and aspartic acid, the protection intermediate of L-glutamic acid are progressively connect saturated fatty alcohol three peptide conjugates of the synthetic protecting group protection of peptide according to existing liquid phase synthetic technology;
(4) N end protecting group and the C that sloughs saturated fatty alcohol three peptide conjugates of protecting group protection successively holds protecting group to obtain target compound.
3. 6 of claim 1 kinds of saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester application in preparation immunosuppressor class medicine.
4. 6 of claim 1 kinds of saturated fatty chain alcohol Glu-Asp-Gly tripeptide esters are as the purposes of the preparation material of preparation micro emulsion or lipidosome drug carrier.
5. 6 of claim 1 kinds of saturated fatty chain alcohol Glu-Asp-Gly tripeptide esters are as the purposes in the targeting preparation material of preparation micro emulsion, lipidosome drug carrier.
CN2009100853198A 2009-05-26 2009-05-26 Saturated fatty chain alcohol Glu-Asp-Gly tripeptide ester, synthetic method and application thereof Pending CN101899085A (en)

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Cited By (6)

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CN102796171A (en) * 2011-05-27 2012-11-28 首都医科大学 Double saturated aliphatic chain alcohol His-Gly-Glu-Asp tetrapeptide esters and preparation method and application thereof
CN102796169A (en) * 2011-05-27 2012-11-28 首都医科大学 Double saturated aliphatic chain alcohol Glu-Asp-Gly-Asp tetrapeptide esters and preparation method and application thereof
CN102796170A (en) * 2011-05-27 2012-11-28 首都医科大学 Double saturated aliphatic chain alcohol His-Gly-Lys-Asp tetrapeptide esters and preparation method and application thereof
CN103450195A (en) * 2012-06-05 2013-12-18 首都医科大学 Tetrahydro-beta-carbolinyl-3-formic acid aliphatic chain alcohol esters, and preparation, nano structure, immunosuppression action and application thereof
CN104211760A (en) * 2013-06-05 2014-12-17 首都医科大学 Conjugates of saturated aliphatic chain alcohol, dexamethasone, and Glu-Asp-Gly, preparation, nano structure, and applications thereof
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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102796171A (en) * 2011-05-27 2012-11-28 首都医科大学 Double saturated aliphatic chain alcohol His-Gly-Glu-Asp tetrapeptide esters and preparation method and application thereof
CN102796169A (en) * 2011-05-27 2012-11-28 首都医科大学 Double saturated aliphatic chain alcohol Glu-Asp-Gly-Asp tetrapeptide esters and preparation method and application thereof
CN102796170A (en) * 2011-05-27 2012-11-28 首都医科大学 Double saturated aliphatic chain alcohol His-Gly-Lys-Asp tetrapeptide esters and preparation method and application thereof
CN103450195A (en) * 2012-06-05 2013-12-18 首都医科大学 Tetrahydro-beta-carbolinyl-3-formic acid aliphatic chain alcohol esters, and preparation, nano structure, immunosuppression action and application thereof
CN103450195B (en) * 2012-06-05 2015-05-20 首都医科大学 Tetrahydro-beta-carbolinyl-3-formic acid aliphatic chain alcohol esters, and preparation, nano structure, immunosuppression action and application thereof
CN104211760A (en) * 2013-06-05 2014-12-17 首都医科大学 Conjugates of saturated aliphatic chain alcohol, dexamethasone, and Glu-Asp-Gly, preparation, nano structure, and applications thereof
CN104211760B (en) * 2013-06-05 2017-05-24 首都医科大学 Conjugates of saturated aliphatic chain alcohol, dexamethasone, and Glu-Asp-Gly, preparation, nano structure, and applications thereof
CN110305030A (en) * 2019-07-01 2019-10-08 昆明理工大学 Amino acid ester hydrochlorides and the preparation method and application thereof
CN110305030B (en) * 2019-07-01 2021-10-15 昆明理工大学 Amino acid ester hydrochloride and preparation method and application thereof

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