Biological reagents and methods for determining the mechanism in the generation of beta-amyloid peptide

A biological reagent defined by formula I below:

• A-B-C I
• wherein:
• A is selected from the group consisting of formulas II, II, IV and V below;
• B is selected from the group consisting of
• a) (CH2CH(R)Q)n
• b) alkylene-Q
• c) substituted alkylene-Q
• wherein R is selected from hydrogen, alkyl, aryl and Q is selected from the group consisting of —O—, —S—, —NH—, —OC(O)NH—, —NHC(O)O—, —NHC(O)NH—, —NHC(O)— and —C(O)NH—;
• and
• C is selected from the group consisting of a solid support and a detectable marker wherein C is optionally linked to Q through a linking arm;
• wherein Formula II is defined as follows:
• of:
• 48
• wherein R11 is selected from the group consisting
• (a) a substituted phenyl group of the formula:
• 49
• wherein
• Rc is selected from the group consisting of acyl, alkyl, alkoxy, alkoxycarbonyl, alkylalkoxy, azido, cyano, halo, hydrogen, nitro, trihalomethyl, thioalkoxy, and wherein Rb and Rc are fused to form a heteroaryl or heterocyclic ring with the phenyl ring wherein the heteroaryl or heterocyclic ring contains from 3 to 8 atoms of which from 1 to 3 are heteroatoms independently selected from the group consisting of oxygen, nitrogen and sulfur;
• Rb and Rb′ are independently selected from the group consisting of hydrogen, halo, nitro, cyano, trihalomethyl, alkoxy, and thioalkoxy with the proviso that Rb, Rb′ and Rc are not all hydrogen and with the further proviso that when Rc is hydrogen, then neither Rb nor Rb′ are hydrogen;
• (b) 2-naphthyl;
• and
• (c) 2-naphthyl substituted at the 4, 5, 6, 7 and/or 8 positions with 1 to 5 substituents selected from the group consisting of alkyl, alkoxy, halo, cyano, nitro, trihalomethyl, and thioalkoxy;
• R12 is selected from the group consisting of hydrogen, alkyl of from 1 to 4 carbon atoms, alkylalkoxy of from 1 to 4 carbon atoms and alkylthioalkoxy of from 1 to 4 carbon atoms;
• and
• R13 is selected from the group consisting of —O—, —S—, —O—(CH2)pC(O)—, —O—(CH2)pC(O)O— and —O—(CH2)pC(O)NH—, wherein p is an integer of from 1 to 2;
• Formula III is defined as follows:
• 50
• wherein R21 is selected from the group consisting of
• a) alkyl, alkenyl, alkcycloalkyl, phenyl-(Rd)m—, naphthyl-(Rd)m— wherein Rd is an alkylene group of from 1 to 8 carbon atoms and m is an integer equal to 0 or 1, cycloalkyl, cycloalkenyl, 3-pyridyl, 4-pyridyl and heteroaryl, other than 3- and 4-pyridyl, of 3 to 10 atoms and 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen wherein the heteroaryl group is optionally substituted with 1 to 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, halo, nitro, thioalkoxy, and thioaryloxy with the proviso that for such heteroaryls when there is at least one nitrogen heteroatom, there is also at least one oxygen and/or sulfur heteroatom;
• (b) a substituted phenyl group of the formula:
• 51
• wherein
• R is alkylene of from 1 to 8 carbon atoms,
• m is an integer equal to 0 or 1,
• Re and Rf are independently selected from the group consisting of hydrogen, hydroxy, fluoro and methyl;
• Rg and Rh are independently selected from the group consisting of hydrogen, alkyl, alkoxy, aryl, cyano, cycloalkyl, halo, heteroaryl, heterocyclic, nitro, trihalomethyl, thioalkoxy, thioaryloxy, thioheteroaryloxy, and —C(O)Rj where Rj is selected from the group consisting of alkyl, aryl, alkoxy and aryloxy;
• and
• Ri is selected from the group consisting of hydrogen, alkyl, aryl, cyano, halo, nitro, and where Rg and Ri are fused to form a methylenedioxy ring with the phenyl ring;
• and
• when Rg and/or Rh and/or Ri is fluoro, chloro, bromo and/or nitro, then Re and/or Rf can also be chloro;
• and
• (c) 1- or 2-naphthyl-(Rk)m— substituted at the 5, 6, 7 and/or 8 positions with 1 to 4 substituents selected from the group consisting alkyl, alkoxy, halo, cyano, nitro, trihalomethyl, and thioalkoxy wherein Rk is an alkylene group of from 1 to 8 carbon atoms and m is an integer equal to 0 or 1;
• R22 is selected from the group consisting of hydrogen, alkyl, phenyl, alkylalkoxy, alkylthioalkoxy;
• X is oxygen or sulfur;
• X′ is hydrogen, hydroxy or fluoro;
• and
• X″ is hydrogen, hydroxy or fluoro, or X′ and X″ together form an oxo group;
• Formula IV is defined as follows:
• 52
• wherein:
• R31 is selected from the group consisting of
• (a) phenyl,
• (b) a substituted phenyl group of the formula:
• 53
• wherein Ro is selected from the group consisting of acyl, alkyl, alkoxy, alkylalkoxy, azido, cyano, halo, hydrogen, nitro, trihalomethyl, thioalkoxy, and wherein Ro and Rm are fused to form a heteroaryl or heterocyclic ring with the phenyl ring,
• Rm and Rn are independently selected from the group consisting of hydrogen, halo, nitro, cyano, trihalomethyl, alkoxy, and thioalkoxy with the proviso that when Ro is hydrogen, then Rm and Rn are either both hydrogen or both substituents other than hydrogen,
• (c) 2-naphthyl,
• (d) 2-naphthyl substituted at the 4, 5, 6, 7 and/or 8 positions with 1 to 5 substituents selected from the group consisting of alkyl, alkoxy, halo, cyano, nitro, trihalomethyl, thioalkoxy, aryl, and heteroaryl,
• (e) heteroaryl, and
• (f) substituted heteroaryl containing 1 to 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, cyano, halo, nitro, heteroaryl, thioalkoxy and thioaryloxy provided that said substituents are not ortho (adjacent) to the heteroaryl attachment to the —NH group;
• R32 is selected from the group consisting of hydrogen, alkyl of from 1 to 4 carbon atoms, alkylalkoxy of from 1 to 4 carbon atoms, alkylthioalkoxy of from 1 to 4 carbon atoms, aryl, heteroaryl, substituted aryl and substituted heteroaryl provided that the substituents are not ortho (adjacent) to the attachment of the aryl or heteroaryl atom to the carbon atom;
• R33 is selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl, heteroaryl, substituted alkyl, substituted alkenyl, substituted alkynyl, and heterocyclic;
• and
• X is selected from the group consisting of oxygen and sulfur;
• and
• Formula V is defined as follows:
• 54
• wherein R41 is selected from the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkenyl, substituted alkynyl, aryl, heteroaryl and heterocyclic;
• R42 is selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, aryl, heteroaryl and heterocyclic;
• each R43 is independently selected from the group consisting of hydrogen and methyl and R43 together with R44 can be fused to form a cyclic structure of from 3 to 8 atoms which is optionally fused with an aryl or heteroaryl group;
• each R44 is independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl, heteroaryl, heterocyclic, substituted alkyl, substituted alkenyl and substituted alkynyl;
• each R45 is selected from hydrogen and methyl or together with R43 forms a cycloalkyl group of from 3 to 6 carbon atoms;
• X is selected from oxygen, sulfur and NH;
• X′ is hydrogen, hydroxy or fluoro;
• and
• X″ is hydrogen, hydroxy or fluoro, or X′ and X″ together form an oxo group;
• Z is selected from the group consisting of a bond covalently linking R41 to —CX′X″—, oxygen and sulfur;
• and
• n is an integer equal to 1 to 3;
• wherein the compounds of formulas II, III, IV and V are effective in inhibiting the cellular release and/or synthesis of β-amyloid peptide.