Method and composition for preparation of H-SAR-LYS-SAR-GLN-NH2

Claim 1

The dipeptide of formula Z-SAR-ε-Z'-LYS-OH, wherein Z and Z' are each selected from the group consisting of (a) ##STR11## wherein R 1 ia phenyl; tolyl; xylyl; adamantyl; allyl; β-cyanoethyl; fluorenylmethyl; benzyl, benzyl wherein the phenyl ring is substituted with from one to three members selected from halo, nitro, loweralkyl, and loweralkoxy; diisopropylmethyl; diphenylmethyl; cyclohexyl; cyclopentyl; vinyl; t-butyl; t-amyl; dimethyltrifluoromethylmethyl; or dimethylbiphenylmethyl; (b) ##STR12## wherein R 2 is loweralkyl of two to four carbons or loweralkyl of one to four carbons substituted with from one to five halo groups; (c) ##STR13## wherein X is S or O and R 3 and R 4 are each benzyl or loweralkyl; (d) ##STR14## wherein R 5 and R 6 taken individually are each loweralkyl or R 5 and R 6 taken together is ##STR15## wherein R 7 and R 8 are each hydrogen or loweralkyl; and (e) ##STR16## wherein R 9 is hydrogen, methyl, halo, or nitro provided that Z is monodentate.

Claim 11

A method for preparation of the peptide H-SAR-LYS-SAR-GLN-NH 2 which comprises: (a) forming Fragment I, which consists of Z-SAR-ε-Z'-LYS-OH, by: (i) protecting the amino group of sarcosine by allowing it to react with a reagent which will introduce the protecting group Z; (ii) protecting the ε-amino group of L-lysine by allowing it to react with a reagent which will introduce the protecting group Z' in such a manner as to specifically protect to ε-amino group; (iii) activating the protected sarcosine formed in step (i) with respect to nucleophilic attack at the carboxy group by an amine to form a carboxy activated protected sarcosine; and (iv) allowing said carboxy activated protected sarcosine to react with the ε-protected L-lysine formed in step (a) (ii) above, whereby Fragment I is formed. (b) forming Fragment II, which consists of H-SAR-GLN-NH 2 , by: (i) preparing Z"-GLN-NH 2 ; (ii) removing the protective group from said Z"-GLN-NH 2 to form unprotected L-glutamine amide; (iii) allowing the unprotected L-glutamine amide to react with the carboxy activated protected sarcosine formed in step (a) (iii) above to form Z-SAR-GLN-NH 2 ; and (iv) removing the protective group from said Z-SAR-GLN-NH 2 whereby Fragment II is formed. (c) connecting Fragment I and Fragment II together to form the protected tetrapeptide Z-SAR-ε-Z'-LYS-SAR-GLN-NH 2 ; (d) removing the protective groups from said protected tetrapeptide; and (e) isolating and purifying the resulting tetrapeptide H-SAR-LYS-SAR-GLN-NH 2 ; wherein Z, Z', and Z" are each selected from the group consisting of: (a) ##STR23## wherein R 1 is phenyl; tolyl; xylyl; adamantyl; allyl; β-cyanoethyl; fluorenylmethyl; benzyl; benzyl wherein the phenyl ring is substituted with from one to three members selected from halo, nitro, loweralkyl, and loweralkoxy; diisopropylmethyl; diphenylmethyl; cyclohexyl; cyclopentyl; vinyl; t-butyl; t-amyl; dimethyltrifluoromethylmethyl; or dimethylbiphenylmethyl; (b) ##STR24## wherein R 2 is loweralkyl of two to four carbons or loweralkyl of one to four carbons substituted with from one to five halo groups; (c) ##STR25## wherein X is S or O and R 3 and R 4 are each benzyl or loweralkyl; (d) ##STR26## wherein R 5 and R 6 taken individually are each loweralkyl or R 5 and R 6 taken together is ##STR27## wherein R 7 and R 8 are each hydrogen or loweralkyl; and (e) ##STR28## wherein R 9 is hydrogen, methyl, halo, or nitro provided that Z is monodentate.

Claim 12

A method for preparation of the peptide H-SAR-LYS-SAR-GLN-NH 2 which comprises: (a) forming Fragment I, which consists of Z-SAR-ε-Z'-LYS-OH, by: (i) protecting the amino group of sarcosine by allowing it to react with a reagent which will introduce the protecting group Z; (ii) protecting the ε-amino group of L-lysine by allowing it to react with a reagent which will introduce the protecting group Z' in such a manner as to specifically protect to ε-amino group; (iii) activating the protected sarcosine formed in step (i) with respect to nucleophilic attack at the carboxy group by an amine to form a carboxy activated protected sarcosine; and (iv) allowing said carboxy activated protected sarcosine to react with the ε-protected L-lysine formed in step (a) (ii) above, whereby Fragment I is formed. (b) forming Fragment II, which consists of H-SAR-GLN-NH 2 , by: (i) preparing Z"-GLN-NH 2 ; (ii) removing the protective group from said Z"-GLN-NH 2 to form unprotected L-glutamine amide; (iii) allowing the unprotected L-glutamine amide to react with the carboxy activated protected sarcosine formed in step (a) (iii) above to form Z-SAR-GLN-NH 2 ; and (iv) removing the protective group from said Z-SAR-GLN-NH 2 whereby Fragment II is formed. (c) connecting Fragment I and Fragment II together to form the protected tetrapeptide Z-SAR-ε-Z'-LYS-SAR-GLN-NH 2 ; (d) removing the protective groups from said protected tetrapeptide; and (e) isolating and purifying the resulting tetrapeptide H-SAR-LYS-SAR-GLN-NH 2 ; wherein Z, Z', and Z" are each selected from the group consisting of: (a) ##STR23## wherein R 1 is phenyl; tolyl; xylyl; adamantyl; allyl; β-cyanoethyl; fluorenylmethyl; benzyl; benzyl wherein the phenyl ring is substituted with from one to three members selected from halo, nitro, loweralkyl, and loweralkoxy; diisopropylmethyl; diphenylmethyl; cyclohexyl; cyclopentyl; vinyl; t-butyl; t-amyl; dimethyltrifluoromethylmethyl; or dimethylbiphenylmethyl; (b) ##STR24## wherein R 2 is loweralkyl of two to four carbons or loweralkyl of one to four carbons substituted with from one to five halo groups; (c) ##STR25## wherein X is S or O and R 3 and R 4 are each benzyl or loweralkyl; (d) ##STR26## wherein R 5 and R 6 taken individually are each loweralkyl or R 5 and R 6 taken together is ##STR27## wherein R 7 and R 8 are each hydrogen or loweralkyl; and (e) ##STR28## wherein R 9 is hydrogen, methyl, halo, or nitro provided that Z is monodentate.

Claim 17

A method for the preparation of the peptide H-SAR-LYS-SAR-GLN-NH 2 which comprises: (a) forming Fragment I, which consists of CBZ-SAR-ε-CBZ-LYS-OH, by: (i) esterifying CBZ-SAR-OH by allowing it to react with an equivalent amount of N-hydroxysuccinimide in the presence of dicyclohexylcarbodiimide to form CBZ-SAR-OSu; and (ii) allowing a slight molar excess CBZ.sup.ε -LYS-OH to react with CBZ-SAR-OSu in the presence of two equivalents of triethylamine to form CBZ-SAR-εCBZ-LYS-OH (Fragment I). (b) forming Fragment II, which consists of H-SAR-GLN-NH 2 , by: (i) preparing CBZ-GLN-NH 2 ; (ii) removing the benzyloxycarbonyl group by catalytic hydrogenation to form H-GLN-NH 2 ; (iii) allowing GLN-NH 2 to react with a molar excess of CBZ-SAR-OSu to form CBZ-SAR-GLN-NH 2 ; and (iv) removing the benzyloxycarbonyl group by catalytic hydrogenation to form H-SAR-GLN-NH 2 (Fragment II). (c) connecting the carboxyl terminus of Fragment I to the amino terminus of Fragment II by mixing equivalent amounts of each fragment in the presence of 1-hydroxybenzotriazole and a slight molar excess of dicyclohexylcarbodiimide; (d) removing the benzyloxycarbonyl groups by catalytic hydrogenation to yield crude product; and (e) isolating and purifying the resulting tetrapeptide H-SAR-LYS-SAR-GLN-NH 2 .

Claim 18

A method for the preparation of the peptide H-SAR-LYS-SAR-GLN-NH 2 which comprises: (a) forming Fragment I, which consists of CBZ-SAR-ε-CBZ-LYS-OH, by: (i) esterifying CBZ-SAR-OH by allowing it to react with an equivalent amount of N-hydroxysuccinimide in the presence of dicyclohexylcarbodiimide to form CBZ-SAR-OSu; and (ii) allowing a slight molar excess CBZ.sup.ε -LYS-OH to react with CBZ-SAR-OSu in the presence of two equivalents of triethylamine to form CBZ-SAR-εCBZ-LYS-OH (Fragment I). (b) forming Fragment II, which consists of H-SAR-GLN-NH 2 , by: (i) preparing CBZ-GLN-NH 2 ; (ii) removing the benzyloxycarbonyl group by catalytic hydrogenation to form H-GLN-NH 2 ; (iii) allowing GLN-NH 2 to react with a molar excess of CBZ-SAR-OSu to form CBZ-SAR-GLN-NH 2 ; and (iv) removing the benzyloxycarbonyl group by catalytic hydrogenation to form H-SAR-GLN-NH 2 (Fragment II). (c) connecting the carboxyl terminus of Fragment I to the amino terminus of Fragment II by mixing equivalent amounts of each fragment in the presence of 1-hydroxybenzotriazole and a slight molar excess of dicyclohexylcarbodiimide; (d) removing the benzyloxycarbonyl groups by catalytic hydrogenation to yield crude product; and (e) isolating and purifying the resulting tetrapeptide H-SAR-LYS-SAR-GLN-NH 2 .

Claim 25

A method for preparation of the peptide H-SAR-LYS-SAR-GLN-NH 2 which comprises: (a) forming Fragment I, which consists of Z-SAR-ε-Z'-LYS-OH, by: (i) protecting the amino group of sarcosine by allowing it to react with a reagent which will introduce the protecting group Z; (ii) protecting the ε-amino group of L-lysine by allowing it to react with a reagent which will introduce the protecting group Z' in such a manner as to specifically protect to ε-amino group; (iii) activating the protected sarcosine formed in step (i) with respect to nucleophilic attack at the carboxy group by an amine to form a carboxy activated protected sarcosine; and (iv) allowing said carboxy activated protected sarcosine to react with the ε-protected L-lysine formed in step (a) (ii) above, whereby Fragment I is formed. (b) forming Fragment II, which consists of H-SAR-GLN-NH 2 , by: (i) preparing Z"-GLN-NH 2 ; (ii) removing the protective group from said Z"-GLN-NH 2 to form unprotected L-glutamine amide; (iii) allowing the unprotected L-glutamine amide to react with a sarcosine N-carboxyanhydride, whereby Fragment II is formed. (c) connecting Fragment I and Fragment II together to form the protected tetrapeptide Z-SAR-ε-Z'-LYS-SAR-GLN-NH 2 ; (d) removing the protective groups from the protected tetrapeptide; and (e) isolating and purifying the resulting tetrapeptide H-SAR-LYS-SAR-GLN-NH 2 ; wherein Z, Z', and Z" are each selected from the group consisting of: (a) ##STR29## wherein R 1 is phenyl; tolyl; xylyl; adamantyl; allyl; β-cyanoethyl; fluorenylmethyl; benzyl; benzyl wherein the phenyl ring is substituted with from one to three members selected from halo, nitro, loweralkyl, and loweralkoxy; diisopropylmethyl; diphenylmethyl; cyclohexyl; cyclopentyl; vinyl; t-butyl; t-amyl; dimethyltrifluoromethylmethyl; or dimethylbiphenylmethyl; (b) ##STR30## wherein R 2 is loweralkyl of two to four carbons or loweralkyl of one to four carbons substituted with from one to five halo groups; (c) ##STR31## wherein X is S or O and R 3 and R 4 are each benzyl or loweralkyl; (d) ##STR32## wherein R 5 and R 6 taken individually are each loweralkyl or R 5 and R 6 taken together is ##STR33## wherein R 7 and R 8 are each hydrogen or loweralkyl; and (e) ##STR34## wherein R 9 is hydrogen, methyl, halo, or nitro provided that Z is monodentate.

Claim 26

A method for preparation of the peptide H-SAR-LYS-SAR-GLN-NH 2 which comprises: (a) forming Fragment I, which consists of Z-SAR-ε-Z'-LYS-OH, by: (i) protecting the amino group of sarcosine by allowing it to react with a reagent which will introduce the protecting group Z; (ii) protecting the ε-amino group of L-lysine by allowing it to react with a reagent which will introduce the protecting group Z' in such a manner as to specifically protect to ε-amino group; (iii) activating the protected sarcosine formed in step (i) with respect to nucleophilic attack at the carboxy group by an amine to form a carboxy activated protected sarcosine; and (iv) allowing said carboxy activated protected sarcosine to react with the ε-protected L-lysine formed in step (a) (ii) above, whereby Fragment I is formed. (b) forming Fragment II, which consists of H-SAR-GLN-NH 2 , by: (i) preparing Z"-GLN-NH 2 ; (ii) removing the protective group from said Z"-GLN-NH 2 to form unprotected L-glutamine amide; (iii) allowing the unprotected L-glutamine amide to react with a sarcosine N-carboxyanhydride, whereby Fragment II is formed. (c) connecting Fragment I and Fragment II together to form the protected tetrapeptide Z-SAR-ε-Z'-LYS-SAR-GLN-NH 2 ; (d) removing the protective groups from the protected tetrapeptide; and (e) isolating and purifying the resulting tetrapeptide H-SAR-LYS-SAR-GLN-NH 2 ; wherein Z, Z', and Z" are each selected from the group consisting of: (a) ##STR29## wherein R 1 is phenyl; tolyl; xylyl; adamantyl; allyl; β-cyanoethyl; fluorenylmethyl; benzyl; benzyl wherein the phenyl ring is substituted with from one to three members selected from halo, nitro, loweralkyl, and loweralkoxy; diisopropylmethyl; diphenylmethyl; cyclohexyl; cyclopentyl; vinyl; t-butyl; t-amyl; dimethyltrifluoromethylmethyl; or dimethylbiphenylmethyl; (b) ##STR30## wherein R 2 is loweralkyl of two to four carbons or loweralkyl of one to four carbons substituted with from one to five halo groups; (c) ##STR31## wherein X is S or O and R 3 and R 4 are each benzyl or loweralkyl; (d) ##STR32## wherein R 5 and R 6 taken individually are each loweralkyl or R 5 and R 6 taken together is ##STR33## wherein R 7 and R 8 are each hydrogen or loweralkyl; and (e) ##STR34## wherein R 9 is hydrogen, methyl, halo, or nitro provided that Z is monodentate.