Therapeutic peptidomimetic macrocycles

A method of selecting a peptidomimetic macrocycle that generates a reduced antibody response capable of being used in therapy to treat a cell proliferative disorder, comprising:

• a) preparing a peptidomimetic macrocycle by forming a cross-linker connecting two amino acid residues of a polypeptide to stabilize a secondary structure of the peptidomimetic macrocycle;
• b) testing the peptidomimetic macrocycle for a reduced antibody response in an in vivo assay as compared to a corresponding uncross-linked polypeptide;
• and
• c) selecting the tested peptidomimetic macrocycle as the peptidomimetic macrocycle that generates the reduced antibody response capable of being used in therapy to treat a cell proliferative disorder if the tested peptidomimetic macrocycle has the reduced antibody response;
• and
• wherein the peptidomimetic macrocycle that generates the reduced antibody response is of Formula (I):
• wherein:
• each A, C, D, and E, is independently a natural or non-natural amino acid;
• B is a natural or non-natural amino acid, amino acid analog,
• [—NH-L3-CO—], [—NH-L3-SO2—], or [—NH-L3-];
• R1 and R2 are independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, each of which except —H is unsubstituted or substituted with halo-;
• R3 is hydrogen, alkyl, alkenyl, alkynyl, arylalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, cycloaryl, or heterocycloaryl, each of which except hydrogen is optionally substituted with R5;
• L is a macrocycle-forming linker of the formula -L1-L2-;
• L1 and L2 and L3 are independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, cycloarylene, heterocycloarylene, or [—R4—K—R4—]n, each being optionally substituted with R5;
• each R4 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, arylene, or heteroarylene;
• each K is independently O, S, SO, SO2, CO, CO2, or CONR3;
• each R5 is independently halogen, alkyl, —OR6, —N(R6)2, —SR6, —SOR6, —SO2R6, —CO2R6, a fluorescent moiety, a radioisotope, or a therapeutic agent;
• each R6 is independently —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heterocycloalkyl, a fluorescent moiety, a radioisotope, or a therapeutic agent;
• R7 is —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, heteroalkyl, cycloalkylalkyl, heterocycloalkyl, cycloaryl, or heterocycloaryl, each of which except —H is optionally substituted with R5, or part of a cyclic structure with a D residue;
• R8 is —H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, heteroalkyl, cycloalkylalkyl, heterocycloalkyl, cycloaryl, or heterocycloaryl, each of which except —H is optionally substituted with R5, or part of a cyclic structure with an E residue;
• each of v and w is independently an integer from 1-1000;
• each of x, y, and z, is independently an integer from 0-10;
• u is an integer from 1-10;
• and
• n is an integer from 1-5.