USRE29732E - Tripeptide - Google Patents
Tripeptide Download PDFInfo
- Publication number
- USRE29732E USRE29732E US05/763,787 US76378777A USRE29732E US RE29732 E USRE29732 E US RE29732E US 76378777 A US76378777 A US 76378777A US RE29732 E USRE29732 E US RE29732E
- Authority
- US
- United States
- Prior art keywords
- sup
- amino
- pro
- gly
- arg
- 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 - Lifetime
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- 229910052739 hydrogen Inorganic materials 0.000 claims abstract description 7
- 239000001257 hydrogen Substances 0.000 claims abstract description 7
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims abstract description 7
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 claims abstract description 7
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims abstract 9
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims abstract 3
- -1 nitro, p-nitrobenzyloxycarbonyl Chemical group 0.000 claims description 17
- 150000001875 compounds Chemical class 0.000 claims description 10
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 claims description 10
- 239000004305 biphenyl Substances 0.000 claims description 3
- 235000010290 biphenyl Nutrition 0.000 claims description 3
- 125000005928 isopropyloxycarbonyl group Chemical group [H]C([H])([H])C([H])(OC(*)=O)C([H])([H])[H] 0.000 claims description 3
- ZUOUZKKEUPVFJK-UHFFFAOYSA-N phenylbenzene Natural products C1=CC=CC=C1C1=CC=CC=C1 ZUOUZKKEUPVFJK-UHFFFAOYSA-N 0.000 claims description 3
- 125000001557 phthalyl group Chemical group C(=O)(O)C1=C(C(=O)*)C=CC=C1 0.000 claims description 3
- UQDJGEHQDNVPGU-UHFFFAOYSA-N serine phosphoethanolamine Chemical compound [NH3+]CCOP([O-])(=O)OCC([NH3+])C([O-])=O UQDJGEHQDNVPGU-UHFFFAOYSA-N 0.000 claims description 3
- 125000002088 tosyl group Chemical group [H]C1=C([H])C(=C([H])C([H])=C1C([H])([H])[H])S(*)(=O)=O 0.000 claims description 2
- 125000001584 benzyloxycarbonyl group Chemical group C(=O)(OCC1=CC=CC=C1)* 0.000 claims 3
- UYWQUFXKFGHYNT-UHFFFAOYSA-N phenylmethyl ester of formic acid Natural products O=COCC1=CC=CC=C1 UYWQUFXKFGHYNT-UHFFFAOYSA-N 0.000 claims 3
- 125000005147 toluenesulfonyl group Chemical group C=1(C(=CC=CC1)S(=O)(=O)*)C 0.000 claims 1
- 125000006239 protecting group Chemical group 0.000 abstract description 8
- 108090000765 processed proteins & peptides Proteins 0.000 abstract description 7
- 230000000903 blocking effect Effects 0.000 abstract description 6
- 108700012941 GNRH1 Proteins 0.000 abstract description 5
- 239000000579 Gonadotropin-Releasing Hormone Substances 0.000 abstract description 5
- XLXSAKCOAKORKW-AQJXLSMYSA-N gonadorelin Chemical compound C([C@@H](C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1[C@@H](CCC1)C(=O)NCC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H]1NC(=O)CC1)C1=CC=C(O)C=C1 XLXSAKCOAKORKW-AQJXLSMYSA-N 0.000 abstract description 5
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- 235000001014 amino acid Nutrition 0.000 description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 6
- 150000001413 amino acids Chemical class 0.000 description 6
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 5
- 239000000543 intermediate Substances 0.000 description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- 125000003275 alpha amino acid group Chemical group 0.000 description 4
- 238000005859 coupling reaction Methods 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- ADFXKUOMJKEIND-UHFFFAOYSA-N 1,3-dicyclohexylurea Chemical compound C1CCCCC1NC(=O)NC1CCCCC1 ADFXKUOMJKEIND-UHFFFAOYSA-N 0.000 description 2
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 2
- 108010016626 Dipeptides Proteins 0.000 description 2
- KRHYYFGTRYWZRS-UHFFFAOYSA-N Fluorane Chemical compound F KRHYYFGTRYWZRS-UHFFFAOYSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 238000010306 acid treatment Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000000921 elemental analysis Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 125000000524 functional group Chemical group 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- MZKZJHTZJCEFKZ-YFKPBYRVSA-N (2s)-n-(2-amino-2-oxoethyl)pyrrolidine-2-carboxamide Chemical compound NC(=O)CNC(=O)[C@@H]1CCCN1 MZKZJHTZJCEFKZ-YFKPBYRVSA-N 0.000 description 1
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 1
- GUVOBXQVANZIKH-ZDUSSCGKSA-N 1-o-tert-butyl 2-o-(4-nitrophenyl) (2s)-pyrrolidine-1,2-dicarboxylate Chemical compound CC(C)(C)OC(=O)N1CCC[C@H]1C(=O)OC1=CC=C([N+]([O-])=O)C=C1 GUVOBXQVANZIKH-ZDUSSCGKSA-N 0.000 description 1
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 102000006771 Gonadotropins Human genes 0.000 description 1
- 108010086677 Gonadotropins Proteins 0.000 description 1
- MRAUNPAHJZDYCK-BYPYZUCNSA-N L-nitroarginine Chemical compound OC(=O)[C@@H](N)CCCNC(=N)N[N+]([O-])=O MRAUNPAHJZDYCK-BYPYZUCNSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 125000000637 arginyl group Chemical group N[C@@H](CCCNC(N)=N)C(=O)* 0.000 description 1
- 238000009903 catalytic hydrogenation reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- BEBCJVAWIBVWNZ-UHFFFAOYSA-N glycinamide Chemical compound NCC(N)=O BEBCJVAWIBVWNZ-UHFFFAOYSA-N 0.000 description 1
- 239000002622 gonadotropin Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 125000001841 imino group Chemical group [H]N=* 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000012442 inert solvent Substances 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 150000002605 large molecules Chemical class 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- KQSSATDQUYCRGS-UHFFFAOYSA-N methyl glycinate Chemical compound COC(=O)CN KQSSATDQUYCRGS-UHFFFAOYSA-N 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000016087 ovulation Effects 0.000 description 1
- 230000027758 ovulation cycle Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 230000006340 racemization Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 125000005931 tert-butyloxycarbonyl group Chemical group [H]C([H])([H])C(OC(*)=O)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/08—Tripeptides
- C07K5/0815—Tripeptides with the first amino acid being basic
- C07K5/0817—Tripeptides with the first amino acid being basic the first amino acid being Arg
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S930/00—Peptide or protein sequence
- Y10S930/01—Peptide or protein sequence
- Y10S930/13—Luteinizing hormone-releasing hormone; related peptides
Definitions
- Gn-RH gonadotropin-relasing hormone
- the formula of the Gn-RH has been identified with the aminoacid sequence pyroGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-NH 2 but in order to make such a large molecule from simple, single aminoacids, a considerable number of steps including several condensation reactions are required. In order to assure such condensations to take place at the desired sites, other active sites or functional groups on the molecule might be conveniently protected by some groups that can be removed at will.
- the present invention is directed to a small peptide chain that contains a blocking group that fulfills the above requirement. It is therefore the main object of the present invention to provide a tripeptide of the formula Y-(N.sup. ⁇ -R')Arg-Pro-Gly-.[.R.]. .Iadd.NH.Iaddend.wherein .[.R represents hydroxy, methoxy or the amino group,.].
- R' is a blocking group that protects the imino group of the arginine moiety and can be removed by a simple chemical step that leaves the aminoacid bonds intact
- Y is hydrogen or a protective group that can be removed by a simple, mild chemical treatment which leaves the remainder of the molecule intact.
- Y is different from hydrogen, it is tert.-butoxycarbonyl (BOC), o-nitrophenylsulfenyl (NPS), 2-(diphenyl)isopropyloxycarbonyl, benzyloxycarbonyl (CBZ) or phthalyl.
- R' may be nitro, p-nitrobenzyloxycarbonyl, tetrachloroisopropyloxyphthaloyl or p-tolylsulfonyl (tos.).
- nitro or tos. groups are preferred because they are removable by a simple treatment with catalytic hydrogenation or hydrofluoric acid. Others mentioned must be removed by more complex reactions.
- the new compounds of the present invention are prepared by reacting BOC-proline p-nitrophenyl ester with glycinamide .[.or glycine methyl ester.]., preferably by using an excess of the latter, and the obtained protected dipeptide is converted to Pro-Gly-.[.R.]. .Iadd.-NH 2 .Iaddend. by a mild acid treatment.
- the free dipeptide is then reacted with BOC-(N.sup. ⁇ -R')-Arg in the presence of dicyclohexylcarbodiimide and an inert solvent. After removing the formed dicyclohexylurea, the mixture is stripped of the solvent and the residue is purified by chromatography.
- the N.sup. ⁇ -BOC group can be removed easily by a mild acid treatment in an inert organic medium, while retaining the blocking group R'. .[.Where R is methoxy, the free acid is obtained by hydrolysis in known manner..].
- a solution of 514 mg. of prolylglycinamide in 8 ml. of pyridine is mixed at room temperature with 619 mg. of dicyclohexylcarbodiimide and 106.2 mg. of N.sup. ⁇ -benzyloxycarbonyl-N.sup. ⁇ -nitroarginine.
- the formed dicyclohexylurea is filtered off and the filtrate is evaporated resulting in an oil.
- This oil is placed on a chromatographic column containing 35 g. of silica gel using 5% methanolic chloroform as the solvent. Elution of the column with 5% methanolic chloroform removes some of the impurities contained in the crude product.
- the pure material is eluted when the methanol concentration is increased to 15%.
- 1.319 g. (87% of theory) of pure CBZ-(N W -NO 2 )Arg-Pro-Gly-NH 2 of undefined melting point is obtained.
- the material produces a correct elemental analysis and its NMR spectrum is consistent with the assigned structure.
- the compounds show [ ⁇ ] D 25 -25.4° (c.-1, DMF).
- the tripeptide is made wherein the CBZ-group is replaced with the BOC-group.
- this material again does not crystallize.
- the new tripeptide is extremely useful as an intermediate for making longer peptide chains, as for instance in Gn-RH and is particularly well suited as a precursor in such a synthesis because of its optical configuration with Pro and Arg both being present in the L-form, and the retention of the protective group in the .[.orginine.]. .Iadd.arginine .Iaddend.moiety during the deblocking of the N.sup. ⁇ -position and during any desired subsequent coupling reactions with other aminoacids. During such deblocking and coupling reactions, the new intermediate is chemically and optically stable, i.e., no racemization takes place.
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Life Sciences & Earth Sciences (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The new blocked tripeptide Y-(Nw -R')Arg-Pro-Gly-R wherein R is .[.hydroxy, methoxy or.]. amino, R' is a suitable blocking group and Y is hydrogen or an easily removable protective group has been found to be a valuable intermediate for the preparation of large peptide chains, such as for instance, the decapeptide Gn-RH.
Description
Recent discovery of the aminoacid sequence of the gonadotropin (Gn)-relasing hormone (RH) has made it highly desirable to produce this substance on a practical scale in a purity sufficient to use the substance therapeutically in instances of hormone deficiences and possibly as a regulating agent for the ovulation cycle in female warm-blooded animals. For instance, it has been found that small doses of Gn-RH, administered by intravenous injections to female sheep in the anestrus cycle, produces ovulation. The formula of the Gn-RH has been identified with the aminoacid sequence pyroGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-NH2 but in order to make such a large molecule from simple, single aminoacids, a considerable number of steps including several condensation reactions are required. In order to assure such condensations to take place at the desired sites, other active sites or functional groups on the molecule might be conveniently protected by some groups that can be removed at will.
A relatively simple method has now been devised to produce the desired aminoacid chain in surprisingly good yields. The new methods involves a minimum of group-protecting and -removal reactions for such protective groups and employs a number of new intermediates which are important stepping stones for making Gn-RH and other peptides.
For the purpose of the present disclosure, it is to be understood that all aminoacids used herein are in their optically active L-form except for glycine.
It has now been found that in order to prepare the decapeptide referred to above, various new intermediates are necessary to accomplish the most practical synthesis for such large peptides. These intermediates require so-called protecting groups on those functional groups that may interfere with the desired coupling reaction that extends the peptide chain to a larger number of aminoacids. Such a protective group has to be found sufficiently strongly to the aminoacid's functional group that it will remain attached thereto when the blocking group at the N.sup.α -position is removed in order to make that site reactive for coupling with a chain-extending aminoacid. By properly selecting these protective groups, other N.sup.α -blocked aminoacids can be attached to the N.sup.α -position of the present polypeptide and all protective groups can be removed at the point where the desired chain is completed.
The present invention is directed to a small peptide chain that contains a blocking group that fulfills the above requirement. It is therefore the main object of the present invention to provide a tripeptide of the formula Y-(N.sup.ω -R')Arg-Pro-Gly-.[.R.]. .Iadd.NH.Iaddend.wherein .[.R represents hydroxy, methoxy or the amino group,.]. R' is a blocking group that protects the imino group of the arginine moiety and can be removed by a simple chemical step that leaves the aminoacid bonds intact, and Y is hydrogen or a protective group that can be removed by a simple, mild chemical treatment which leaves the remainder of the molecule intact. More specifically, where Y is different from hydrogen, it is tert.-butoxycarbonyl (BOC), o-nitrophenylsulfenyl (NPS), 2-(diphenyl)isopropyloxycarbonyl, benzyloxycarbonyl (CBZ) or phthalyl. R' may be nitro, p-nitrobenzyloxycarbonyl, tetrachloroisopropyloxyphthaloyl or p-tolylsulfonyl (tos.). Among these, nitro or tos. groups are preferred because they are removable by a simple treatment with catalytic hydrogenation or hydrofluoric acid. Others mentioned must be removed by more complex reactions.
In a simple embodiment, the new compounds of the present invention are prepared by reacting BOC-proline p-nitrophenyl ester with glycinamide .[.or glycine methyl ester.]., preferably by using an excess of the latter, and the obtained protected dipeptide is converted to Pro-Gly-.[.R.]. .Iadd.-NH2 .Iaddend. by a mild acid treatment. The free dipeptide is then reacted with BOC-(N.sup.ω -R')-Arg in the presence of dicyclohexylcarbodiimide and an inert solvent. After removing the formed dicyclohexylurea, the mixture is stripped of the solvent and the residue is purified by chromatography. The N.sup.α -BOC group can be removed easily by a mild acid treatment in an inert organic medium, while retaining the blocking group R'. .[.Where R is methoxy, the free acid is obtained by hydrolysis in known manner..].
In order to illustrate the method for obtaining the compounds of the present invention, reference is made to the following examples which are, however, not to be interpreted as limiting the scope of this invention in any respect.
A solution of 514 mg. of prolylglycinamide in 8 ml. of pyridine is mixed at room temperature with 619 mg. of dicyclohexylcarbodiimide and 106.2 mg. of N.sup.α -benzyloxycarbonyl-N.sup.ω -nitroarginine. After 16 hours, the formed dicyclohexylurea is filtered off and the filtrate is evaporated resulting in an oil. This oil is placed on a chromatographic column containing 35 g. of silica gel using 5% methanolic chloroform as the solvent. Elution of the column with 5% methanolic chloroform removes some of the impurities contained in the crude product. The pure material is eluted when the methanol concentration is increased to 15%. By combining the appropriate fractions and evaporation of the solvent, 1.319 g. (87% of theory) of pure CBZ-(NW -NO2)Arg-Pro-Gly-NH2 of undefined melting point is obtained. The material produces a correct elemental analysis and its NMR spectrum is consistent with the assigned structure. The compounds show [α]D 25 -25.4° (c.-1, DMF).
Similarly, the tripeptide is made wherein the CBZ-group is replaced with the BOC-group. However, this material again does not crystallize.
.[.When the above Pro-Gly-NH2 is replaced by an equimolar amount of Pro-Gly-OCH3, the same reaction sequence yields the N.sup.α,N.sup.ω -diprotected Arg-Pro-Gly-OCH3 which is hydrolyzed at room temperature in 6 hours with one molar equivalent of 1 N aqueous sodium hydroxide using a mixture of dimethylformamide/dioxan 1:1 as the solvent for the diprotected Arg-Pro-Gly-OCH3 to Arg-Pro-Gly-OH carrying the selected blocking groups in the N.sup.α -positions of Arg..].
A solution of 1.013 g. of CBZ-(N.sup.ω -NO2)Arg-Pro-Gly-NH2 from Example 1 in 8 ml. of acetic acid is treated with 8 ml. of 32% hydrobromic acid in acetic acid. After one hour, the solution is added to ether and the precipitate is separated, washed five times by suspending it in ether and decanting the supernatant from the solid. The solid is then treated in methanol with an ion exchange resin in its OH-form and the resulting suspension is filtered. The resin is washed with 10% acetic acid in methanol and the combined wash liquor and filtrate is evaporated to a solid of undefined melting point. The elemental analysis confirms the expected structure (N.sup.ω -NO2)Arg-Pro-Gly-NH2 which shows a single spot on TLC with Rf 0.15 in 15% methanol/chloroform.
By replacing (CBZ)-(N.sup.ω -NO2)Arg-Pro-Gly-NH2 with the corresponding BOC- protected tripeptide amide or the corresponding N.sup.ω -tos. analogues from Example 1, the above procedure yields (N.sup.ω -NO2)Arg-Pro-Gly-NH2 or (N.sup.ω -tos.)Arg-Pro-Gly-NH2, respectively. In all instances, the N.sup.α -deprotection step produces a yield of >90% of theory.
The new tripeptide is extremely useful as an intermediate for making longer peptide chains, as for instance in Gn-RH and is particularly well suited as a precursor in such a synthesis because of its optical configuration with Pro and Arg both being present in the L-form, and the retention of the protective group in the .[.orginine.]. .Iadd.arginine .Iaddend.moiety during the deblocking of the N.sup.α -position and during any desired subsequent coupling reactions with other aminoacids. During such deblocking and coupling reactions, the new intermediate is chemically and optically stable, i.e., no racemization takes place.
Claims (7)
1. The optically active L-form of the tripeptide Y-(N.sup.ω -R')Arg-Pro-Gly-R wherein R is .[.hydroxy, methoxy or.]. amino, R' is nitro, p-nitrobenzyloxycarbonyl, tetrachloroisopropyloxyphthaloyl or p-tolylsulfonyl, and wherein Y is hydrogen, tert.-butoxycarbonyl, o-nitrophenylsulfenyl, 2-(diphenyl)isopropyloxycarbonyl, benzyloxycarbonyl or phthalyl.
2. The compounds of claim 1 wherein R is .[.hydroxy, methoxy or.]. amino, R' is p-toluenesulfonyl, p-nitrobenzyloxycarbonyl, tetrachloroisopropoxyphthaloyl, or nitro and Y is hydrogen, tert.-butoxycarbonyl, o-nitrophenylsulfenyl, phthalyl, benzoyloxycarbonyl or 2-(diphenyl)isopropyloxycarbonyl.
3. The compound of claim 2 wherein R is amino, R' is nitro and Y is hydrogen.
4. The compound of claim 2 wherein R is amino, R' is p-toluenesulfonyl and Y is hydrogen.
5. The compound of claim 2 wherein R is amino, R' is p-toluenesulfonyl and Y is benzyloxycarbonyl.
6. The compound of claim 2 wherein R is amino, R' is toluenesulfonyl and Y is tert.-butoxycarbonyl.
7. The compound of claim 2 wherein R is amino, R' is nitro and Y is benzyloxycarbonyl. .[.8. The compound of claim 2 wherein R is amino, R' is nitro and Y is trert.-butoxycarbonyl. .].
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US23787772A | 1972-03-24 | 1972-03-24 |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| US23787772A Reissue | 1972-03-24 | 1972-03-24 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| USRE29732E true USRE29732E (en) | 1978-08-15 |
Family
ID=22895618
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| US00237877A Expired - Lifetime US3781272A (en) | 1972-03-24 | 1972-03-24 | Tripeptide |
| US05/763,787 Expired - Lifetime USRE29732E (en) | 1972-03-24 | 1977-01-31 | Tripeptide |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| US00237877A Expired - Lifetime US3781272A (en) | 1972-03-24 | 1972-03-24 | Tripeptide |
Country Status (1)
| Country | Link |
|---|---|
| US (2) | US3781272A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5238921A (en) | 1990-02-27 | 1993-08-24 | Agency Of Industrial Science And Technology | Oligopeptide, angiotensin converting enzyme inhibitors, hypotensive agent, and method for treatment of hypertension |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3953416A (en) * | 1971-12-20 | 1976-04-27 | Karl Folkers | Synthetic decapeptide having the activity of the luteinizing hormone releasing hormone and method for manufacturing the same |
| US4128540A (en) * | 1972-05-25 | 1978-12-05 | Parke, Davis & Company | Pyroglutamyl-histidyl-tryptophanyl-seryl-tyrosyl hydrazides |
| US5707965A (en) * | 1974-11-14 | 1998-01-13 | Intesco Laboratories, Inc. | Peptides for control of intestinal motility |
-
1972
- 1972-03-24 US US00237877A patent/US3781272A/en not_active Expired - Lifetime
-
1977
- 1977-01-31 US US05/763,787 patent/USRE29732E/en not_active Expired - Lifetime
Non-Patent Citations (1)
| Title |
|---|
| guttmann et al., Helv. Chem. Acta, vol. 44 (1961), pp. 1713-1723. * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5238921A (en) | 1990-02-27 | 1993-08-24 | Agency Of Industrial Science And Technology | Oligopeptide, angiotensin converting enzyme inhibitors, hypotensive agent, and method for treatment of hypertension |
Also Published As
| Publication number | Publication date |
|---|---|
| US3781272A (en) | 1973-12-25 |
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