Rapamycin conjugates

1. A rapamycin conjugate represented by formula I, or a salt thereof:

wherein R 1 and R 2 are each, independently, hydrogen or —(R 3 -L-R 4 ) a —;

L is a linking group;

R 3 is selected from the group consisting of carbonyl, —S(O)—, —S(O) 2 , —P(O) 2 —, —P(O)(CH 3 )—, —C(S)—, and —CH 2 C(O)—;

R 4 is a selected from the group consisting of carbonyl, —NH—, —S—, —CH 2 —, and —O—;

a=1-5;

x=0-1;

y=0-1;

z is from about 1 to about 120;

and Carrier is immunogenic carrier material, detector carrier material, or a solid matrix,

with the proviso that R 1 and R 2 are both not hydrogen; and further provided that when a is greater than 1, each L group can be the same or different; and still further provided that x is 0 if R 1 is hydrogen and y is 0 if R 2 is hydrogen, and if x and y are both 1, the Carrier moiety is the same in both cases.

2. A rapamycin conjugate represented by formula II, or a salt thereof:

wherein R 1 and R 2 are each, independently, hydrogen or —R 3 -L-R 4 —;

L is -A-(CR 5 R 6 ) b [B—(CR 7 R 8 ) d ] e —;

A is —CH 2 — or —NR 9 —;

B is —O—, —NR 9 —, —S—, —S(O)—, or —S(O) 2 —;

R 3 is selected from the group consisting of carbonyl, —S(O)—, —S(O) 2 —, —P(O) 2 —, —P(O) 2 (CH 3 )—, —C(S)—, and —CH 2 C(O)—;

R 4 is selected from the group consisting of carbonyl, —NH—, —S—, —CH 2 —, and —O—;

R 5 , R 6 , R 7 , and R 8 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, halo, hydroxy, trifluoromethyl, arylalkyl of 7-10 carbon atoms, aminoalkyl of 1-6 carbon atoms, hydroxyalkyl of 1-4 carbon atoms, alkoxy of 1-6 carbon atoms, carbalkoxy of 2-7 carbon atoms, cyano, amino, -C02H, or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from the group consisting of alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, and —CO 2 H;

R 9 is hydrogen, alkyl of 1-6 carbon atoms, or aralkyl of 7-10 carbon atoms;

b=0-10;

d=0-10;

e=0-2;

x=0-1;

y=0-1;

z is from about 1 to about 120;

and Carrier is immunogenic carrier material, detector carrier material, or a solid matrix,

with the proviso that R 1 and R 2 are both not hydrogen; and further provided that when b is greater than 1, each of the CR 5 R 6 groups can be the same or different, and when d is greater than 1, each of the CR 7 R 8 groups can be the same or different; and still further provided that x is 0 if R 1 is hydrogen and y is 0 if R 2 is hydrogen, and if x and y are both 1, the Carrier moiety is the same in both cases.

3. A rapamycin conjugate represented by formula III, or a salt thereof:

wherein R 1 and R 2 are each, independently, hydrogen or —(R 3 -L 1 -R 4 ) f —(R 10 -L 2 -R 11 ) g —;

L 1 is —(CH 2 ) h —CHR 12 —(CH 2 ) j —;

L 2 is —(CH 2 ) k -D-(CH 2 ) m -E-;

D is —CH 2 —, —S—S—, or

E is —CH 2 — or —C—;

R 3 and R 10 are each, independently, selected from the group consisting of carbonyl, —S(O)—, —S(O) 2 —, —P(O) 2 —, —P(O)(CH 3 )—, —C(S)—, and —CH 2 C(O)—;

R 4 and R 11 are each, independently, selected from the group consisting of carbonyl, —NH—, —S—, —CH 2 —, and —O—;

R 12 is hydrogen, alkyl of 1-6 carbon atoms, arylalkyl of 7-10 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, —(CH 2 ) n CO 2 R 13 , —(CH 2 ) p NR 14 R 15 , carbamylalkyl of 2-3 carbon atoms, aminoalkyl of 1-4 carbon atoms, hydroxyalkyl of 1-4 carbon atoms, guanylalkyl of 2-4 carbon atoms, mercaptoalkyl of 1-4 carbon atoms, alkylthioalkyl of 2-6 carbon atoms, indolylmethyl, hydroxyphenylmethyl, imidazoylmethyl, halo, trifluoromethyl, or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from the group consisting of alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, and —CO 2 H;

R 14 and R 15 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, or arylalkyl of 7-10 carbon atoms;

R 13 is hydrogen, alkyl of 1-6 carbon atoms, arylalkyl of 7-10 carbon atoms, alkenyl of 2-7 carbon atoms, alkynyl of 2-7 carbon atoms, or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from the group consisting of alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, and —CO 2 H;

f=0-3;

g=0-1;

h=0-10;

j=0-10;

k=0-10;

m=0-10;

n=0-6;

p=0-6;

x=0-1;

y=0-1;

z is from about 1 to about 120;

and Carrier is immunogenic carrier material, detector carrier material, or a solid matrix, with the proviso that R 1 and R 2 are both not hydrogen; and further provided that f and g are both not 0 and when f is greater than 1, each of the —(R 3 -L 1 -R 4 )— moieties can be the same or different; and still further provided that x is 0 if R 1 is hydrogen and y is 0 if R 2 is hydrogen, and if x and y are both 1, the Carrier moiety is the same in both cases.

4. The rapamycin conjugate of claim 1 , wherein said immunogenic carrier material is selected from the group consisting of keyhole limpet hemocyanin and ovalbumin.

5. The rapamycin conjugate of claim 2 , wherein said immunogenic carrier material is selected from the group consisting of keyhole limpet hemocyanin and ovalbumin.

6. The rapamycin conjugate of claim 2 , wherein said immunogenic carrier material is selected from the group consisting of keyhole limpet hemocyanin and ovalbumin.

7. An immunoassay kit for measuring the blood level of a rapamycin comprising a conjugate of any one of claims 1 , 2 or 3 , wherein the carrier is a detector carrier material.

8. The immunoassay kit of claim 1 , wherein said detector carrier material is horseradish peroxidase.

9. The rapamycin conjugate of claim 1 , wherein in formula I, R 1 and R 2 are each —(R 3 -L-R 4 ) a —; R 3 is carbonyl; L is a CH 2 CH 2 linking group; R 4 is a carbonyl, a is 1, x is 1, and y is 0.

10. The conjugate of claim 3 , wherein in formula III, R 1 is (R 3 -L 1 -R 4 ) f —(R 10 -L 2 -R 11 ) g —, R 2 is hydrogen, R 3 is carbonyl; R 4 is NH, and h is 0, R 12 is hydrogen, j is 0, and f is 1; R 10 is carbonyl is 0, D is

m is 3, E is

and R 11 is NH, g is 1, x is 1 and y is 0, and Carrier is horseradish peroxidase.

11. A conjugate represented by formula III

wherein R 1 is (R 3 -L 1 -R 4 ) f —(R 10 -L 2 -R 11 ) g —, R 2 is hydrogen, R 3 is carbonyl, f is 1 and L 1 is (CH 2 ) h —CHR 12 —(CH 2 ) j , h is 0, R 12 is hydrogen, j is 0; R 4 is NH, R 10 is carbonyl; L 2 is (CH 2 ) k -D-(CH 2 ) m -E, k is 1, D is

m is 0, E is carbonyl, R 11 is NH, g is 1, x is 1 and y is 0, and Carrier is horseradish peroxidase.