Patent GB-1516725-A (GB)
Title: Polypeptides having acth-like action
IPC: C07K14/575
Filed: 1975-07-11
Abstract: 1516725 Polypeptides having ACTH activity SHIONOGI & CO Ltd 11 July 1975 [30 July 1974 29298/75 Heading C3H [Also in Division C2] Per se polypeptides (I), having ACTH-activity of formula X1 - Tyr - Ser - X2 - X3 - His - Phe - Arg - Trp - Gly - Lys - Pro - Val - Gly - (Lys)n - Y wherein X1 is an alpha-aminoisobutyric acid, #- alanine, L-serine, D-serine, glycine, D-alanine, sarcosine or gamma-aminobutyric acid residue; X2 is a L-methionine, L-norleucine, L-isoleucine, or L-norvaline residue; X3 is a L-glutamic acid or L-glutamine residue; n is an integer of from 5 to 10; Y is R1, NR2R3 or NH(CH2)m-NR4R5 wherein R1 is hydroxy, or alkoxy having from 1 to 5 carbon atoms; R2, R3, R4, R5 are independently hydrogen or alkyl having from 1 to 5 carbon atoms; m is an integer of from 1 to 10; Y being a group which is bound to the carbonyl group of the C-terminal lysine residue. The peptides (I) may be prepared by condensation of the constituent amino acids and/or peptides via the azide method, the dicyclohexylcarbodiimide method, the mixed anhydride method, the carbonyldiimidazole method, the activated ester method, the isoxazolium method, the N-carboxyanhydride method, solid phase peptide synthesis or the oxidation-reduction method, optionally in the presence of N- hydroxysuccinimide, N-hydroxy-5-norbornene- 2,3-dicarboxyimide or 1-hydroxybenzotriazole. Functional groups which do not participate in peptide bond formation are optionally protected, and protective groups are removed by catalytic hydrogenolysis, acid solvolysis, hydrolysis, hydrazinolysis or sodium in liquid ammonia, preferably in an inert solvent. The condensation and deprotection may be effected at - 20 C. to 60 C. The impure peptide may be purified by ion-exchange chromatography, partition chromatography, exclusion chromatography, countercurrent chromatography or adsorption chromatography. Polypeptides (I) may be in the form of an acid addition salt, or a complex which form with heavy metal ions or polyamino acids. Examples describe the preparation of: (Aibl, Lys<17,18,19>)-ACTH(1-19)-NH2 (X1=H-Aib, X2=Met, X3=Glu, n=5, Y= NH2); (Aib<1>, Lys<17,18,19,20>)-ACTH(1-20)- NH2 (X1 = H-Aib, X2=Met, X3=Glu, n=6, Y=NH2). Polypeptide intermediates isolated are: EICOSAPEPTIDE: Boc-Aib-Tyr-Ser-Met- Glu(OBu<t>) - His - Phe - Arg - Trp - Gly - Lys (Boc) - Pro - Val Gly - Lys(Boc) - Lys(Boc) - Lys(Boc) - Lys(Boc) -Lys-(Boc)-NH2. NONADECAPEPTIDE: Boc - Aib - Tyr - Ser - Met - Glu (OBu<t>) His - Phe - Arg - Trp - Gly - Lys - Pro Val - Gly - Lys (Boc) Lys (Boc)- Lys (Boc) - Lys-(Boc)-NH2. DECAPEPTIDES: Boc - - Aib Tyr - Ser - Met Glu(OBu<t>) - His - Phe - Arg - Trp - Gly - OH HCl; H - Lys(Boc) - Pro - Val - Gly - Lys (Boc) - Lys(Boc) - Lys(Boc) - Lys(Boc) - Lys (Boc) - Lys(Boc) - NH2. NONAPEPTIDES: Z and H - Lys(Boc) - Pro- Val - Gly - Lys(Boc) - Lys(Boc) - Lys(Boc) - Lys (Boc) - - Lys(Boc) - NH2. HEXAPEPTIDES: Z and H-Lys(Boc)-Lys (Boc) - - Lys (Boo) - Lys(Boc) - Lys(Boc) - Lys(Boc)- NH2. PENTAPEPTIDES: Z and H-Lys(Boc)-Lys (Boc) - - Lys(Boc) - Lys(Boc) - Lys(Boc) - NH2. TETRAPEPTIDES: Z and H-Lys(Boc)- Lys(Boc) - Lys(Boc) - Lys(Boc) - NH2. TRIPEPTIDES: Z and H-Lys(Boc) - Lys (Boc) - - Lys(Boc) - NH2. Dipeptide intermediates isolated are: Z and H-Lys(Boc)-Lys(Boc)-NH2. Medicinal compositions comprise the polypeptides (I), or an acid addition salt thereof, or a heavy metal ion complex or polyamino acid complex thereof, together with a pharmaceutically acceptable or veterinarily acceptable diluent, carrier or excipient therefor, which may be used for the induction of adrenalstimulating activity; for testing the adrenocortical function; or for the treatment of inflammations, adrenal insufficiency due to pituitary disorder, acute or chronic articular rheumatisms, allergic diseases or adrenarches; in a non-human animal.