Temporary minimal protection synthesis of serine-containing polypeptides
AssigneeSyntex (U.S.A.) Inc.
InventorsJohn J. Nestor, Jr., Natalie L. McClure, Humberto Arzeno
Life of this patent
Estimating the term…
Abstract
In a process for the solid phase synthesis of a polypeptide containing at least one serine residue, the improvement comprising temporarily protecting the side chain of the serine residue with a protecting group which is removed immediately following the addition of the serine to the peptide chain.
Claims19
In a process for the solid phase synthesis of a LH-RH analog having at least one serine, the improvement comprising temporarily protecting the side-chain of each serine residue with a protecting group labile to N.sup..alpha. -deprotecting agents wherein the serine side-chain protecting group and the N.sup..alpha. -protecting group are removed immediately following the addition of each serine to the polypeptide chain, prior to adding the next amino acid in the sequence.
In a process for the solid-phase synthesis of a compound having an amino acid sequence of the formula
- R.sup.1 -R.sup.2 -R.sup.3 -Ser-Tyr-R.sup.4 -Leu-R.sup.5 -Pro-R.sup.6(I)
- wherein
- R.sup.1 is selected from (pyro)Glu and N-Ac-D-Nal(2);
- R.sup.2 is selected from His, D-p-Cl-Phe and D-p-F-Phe;
- R.sup.3 is selected from Trp, D-Trp, D-Nal(2) and D-Pal(3);
- R.sup.4 is selected from D-Nal(2), D-hArg(Et).sub.2, D-Lys(iPr), D-hArg(Bu), D-hArg(CH.sub.2,CF.sub.3).sub.2, D-His(Bzl), D-Leu, D-Pal(3), D-Ser(tBu) and D-Trp;
- R.sup.5 is selected from Arg, hArg(Et).sub.2, hArg(Bu), hArg(CH.sub.2, CF.sub.3).sub.2 and Lys(iPr);
- and
- R.sup.6 is selected from Gly-NH.sub.2, NH-NHCONH.sub.2, D-Ala-NH.sub.2 and NHEt;
- wherein the amino acids are provided with N.sup..alpha. protection;
- the improvement comprising temporarily protecting the side chain of serine at position 4 with a side chain protecting group labile to N.sup..alpha. -deprotecting agents, wherein the serine side chain protecting group and the N.sup..alpha. -protecting group are removed immediately following the addition of the serine to the polypeptide chain, prior to addition of the next amino acid in the sequence.
A process for the solid phase synthesis of nafarelin comprising successively coupling of an inert solid support the following N.sup..alpha. protected amino acids:
- N.sup..alpha. -Boc-Gly
- N.sup..alpha. -Boc-Pro
- N.sup..alpha. -Boc-Arg.HCl
- N.sup..alpha. -Boc-Leu
- N.sup..alpha. -Boc-D-Nal(2)
- N.sup..alpha. -Boc-Tyr
- N.sup..alpha. -Boc-Ser(protected)
- N.sup..alpha. -Boc-Trp
- N.sup..alpha. -Boc-His(protected)
- N.sup..alpha. -Boc-(pyro)Glu,
- treating the product between each step-wise addition with a solution of HCl/iPrOH/CH.sub.2 Cl.sub.2 at temperatures of 0.degree. C. to 45.degree. C. and then cleaving the nafarelin from the support by treatment with a saturated solution of ammonia at a temperature of 10.degree. to 50.degree. C.
A process for the solid phase synthesis of ganirelix comprising successively coupling on an inert solid support the following N.sup..alpha. protected amino acids,
- N.sup..alpha. -Boc-D-Ala;
- N.sup..alpha. -Boc-Pro;
- N.sup..alpha. -Boc-hArg(Et).sub.2.HCl;
- N.sup..alpha. -Boc-Leu;
- N.sup..alpha. -Boc-D-hArg(Et).sub.2.HCl;
- N.sup..alpha. -Boc-Tyr;
- N.sup..alpha. -Boc-Ser(protected);
- N.sup..alpha. -Boc-D-Pal(3);
- N.sup..alpha. -Boc-D-p-Cl-Phe;
- N.sup..alpha. -Boc-Ac-D-Nal(2);
- treating the product between each step-wise addition with a solution of HCl/iPrOH/CH.sub.2 Cl.sub.2 at temperatures of 0.degree. C. to 45.degree. C. and then cleaving the ganirelix from the support by treatment with a saturated solution of ammonia at a temperature of 10.degree. to 50.degree. C.
Dependencies are inferred from the claim text.