1. A compound of claim having a structural Formula (IIIa) or (IIIa’),
or a tautomer, andor salt thereof, wherein:
A is \u2014NH2;
B is hydrogen, alkyl, \u2014CN, \u2014OR4, \u2014S(O)aR4, \u2014NR4R5, \u2014C(O)NR4R5, \u2014CO2R4, \u2014NR4CO2R5, \u2014NR4C(O)NR5R6, \u2014NR4C(S)NR5R6, \u2014NR4C(\u2550NH)NR5R6, \u2014SO2NR4R5, \u2014NR4SO2R5, \u2014NR4SO2NR5R6, \u2014B(OR4)(OR5), \u2014P(O)(OR4)(OR5), or \u2014P(O)(R4)(OR5);
C is \u2014CO2R7;
a is 0, 1 or 2;
R4, R5, R6, and R7 are independently hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl or substituted heteroarylalkyl; or alternatively, R4 and R5, or R5 and R6, together with the atoms to which they are bonded, form a cycloheteroalkyl or substituted cycloheteroalkyl ring
R21 is hydrogen;
R22 is hydrogen, alkyl, substituted alkyl, halo, \u2014CN, or \u2014OR27;
R23 is hydrogen, alkyl, substituted alkyl, \u2014CN, \u2014OR29, \u2014S(O)fR29, \u2014OC(O)R29, \u2014NR29R30, \u2014C(O)NR29R30, \u2014C(O)R29, \u2014CO2R29, \u2014SO2NR29R30, or \u2014NR29SO2R30;
R24 is \u2014O-L1-L2-R33 or \u2014OR33;
L1 is alkylene or substituted alkylene;
L2 is \u2014NR34\u2014,\u2014O\u2014, \u2014S\u2014, \u2014NR34\u2014C(O)\u2014, \u2014C(O)\u2014NR34\u2014, \u2014O\u2014C(O)\u2014, \u2014C(O)\u2014O\u2014, \u2014NR34\u2014C(O)\u2014O\u2014, \u2014O\u2014C(O)\u2014NR34\u2014,\u2014NR34\u2014C(O)\u2014NR35\u2014, \u2014O\u2014C(O)\u2014O\u2014,-heterocyclylene-C(O)\u2014, or -(substituted heterocyclylene)-C(O)\u2014;
R33 is alkyl, substituted alkyl, carbocyclyl, substituted carbocyclyl; aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocyclyl, substituted heterocyclvl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, or substituted heteroarylalkyl;
R34 and R35 are independently hydrogen, alkyl, substituted alkyl, carbocyclyl, substituted carbocyclyl; aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocyclyl, substituted heterocyclyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, or substituted heteroarylalkyl;
f and g are independently 0, 1 or 2; and
R27, R29, and R30 are independently hydrogen, or alkyl;
wherein substituted, when used to modify a specified group or radical, means that one or more hydrogen atoms of the specified group or radical are each independently replaced with one or more substituent group(s);
wherein substituent groups on saturated carbon atoms in the substituted group or radical include one or more of \u2014Ra, halo, \u2014O\u2212, \u2550O, \u2014ORb, \u2014SRb, \u2014S\u2212, \u2550S, \u2014NRcRc, \u2550NRb, \u2550N\u2014ORb, trihalomethyl, \u2014CF3, \u2014CN, \u2014OCN, \u2014SCN, \u2014NO, \u2014NO2, \u2550N2, \u2014N3, \u2014S(O)2Rb, \u2014S(O)2NRb, \u2014S(O)2O\u2212, \u2014S(O)2ORb, \u2014OS(O)2Rb, \u2014OS(O)2O\u2212, \u2014OS(O)2ORb, \u2014P(O)(O\u2212)2, \u2014P(O)(ORb)(O\u2212), \u2014P(O)(ORb)(ORb), \u2014C(O)Rb, \u2014C(S)Rb, \u2014C(NRb)Rb, \u2014C(O)O\u2212, \u2014C(O)ORb, \u2014C(S)ORb, \u2014C(O)NRcRc, \u2014C(NRb)NRcRc, \u2014OC(O)Rb, \u2014OC(S)Rb, \u2014OC(O)O\u2212, \u2014OC(O)ORb, \u2014OC(S)ORb, \u2014NRbC(O)Rb, \u2014NRbC(S)Rb, \u2014NRbC(O)O\u2212, \u2014NRbC(O)ORb, \u2014NRbC(S)ORb, \u2014NRbC(O)NRcRc, \u2014NRbC(NRb)Rb or \u2014NRbC(NRb)NRcRc,
wherein each Ra is independently selected from the group consisting of alkyl, cycloalkyl, heteroalkyl, cycloheteroalkyl, aryl, arylalkyl, heteroaryl and heteroarylalkyl; each Rb is independently hydrogen or Ra; and each Rc is independently Rb or alternatively, the two Rcs may be taken together with the nitrogen atom to which they are bonded form a 4-, 5-, 6- or 7-membered cycloheteroalkyl which may optionally include from 1 to 4 of the same or different additional heteroatoms selected from the group consisting of O, N and S;
wherein the one or more substituent groups, taken together with the atoms to which they are bonded, may form a cyclic ring including cycloalkyl and cycloheteroalkyl;
wherein substituent groups on unsaturated carbon atoms in the specified group or radical include \u2014Ra, halo, \u2014O\u2212, \u2014ORb, \u2014SRb, \u2014S\u2212, \u2014NRcRc, trihalomethyl, \u2014CF3, \u2014CN, \u2014OCN, \u2014SCN, \u2014NO, \u2014NO2, \u2014N3, \u2014S(O)2,Rb, \u2014S(O)2O\u2212, \u2014S(O)2ORb, \u2014OS(O)2Rb, \u2014OS(O)2O\u2212, \u2014OS(O)2ORb, \u2014P(O)(O\u2212)2, \u2014P(O)(ORb)(O\u2212), \u2014P(O)(ORb)(ORb), \u2014C(O)Rb, \u2014C(S)Rb, \u2014C(NRb)Rb, \u2014C(O)O\u2212, \u2014C(O)ORb, \u2014C(S)ORb, \u2014C(O)NRcRc, \u2014C(NRb)NRcRc, \u2014OC(O)Rb, \u2014OC(S)Rb, \u2014OC(O)O\u2212, \u2014OC(O)ORb, \u2014OC(S)ORb, \u2014NRbC(O)Rb, \u2014NRbC(S)Rb, \u2014NRbC(O)O\u2212, \u2014NRbC(O)ORb, \u2014NRbC(S)ORb, \u2014NRbC(O)NRcRc, \u2014NRbC(NRb)Rb or \u2014NRbC(NRb)NRcRc;
wherein substituent groups on nitrogen atoms in heteroatom-containing groups include \u2014Ra, \u2014O\u2212, \u2014ORb, \u2014SRb, \u2014S\u2212, \u2014NRcRc, trihalomethyl, \u2014CF3, \u2014CN, \u2014NO, \u2014NO2, \u2014S(O)2Rb, \u2014S(O)2O\u2212, \u2014S(O)2ORb, \u2014OS(O)2Rb, \u2014OS(O)2O\u2212, \u2014OS(O)2ORb, \u2014P(O)(O\u2212)2, \u2014P(O)(ORb)(O\u2212), \u2014P(O)(ORb)(ORb), \u2014C(O)Rb, \u2014C(S)Rb, \u2014C(NRb)Rb, \u2014C(O)ORb, \u2014C(S)ORb, \u2014C(O)NRcRc, \u2014C(NRb)NRcRc, \u2014OC(O)Rb, \u2014OC(S)Rb, \u2014OC(O)ORb, \u2014OC(S)ORb, \u2014NRbC(O)Rb, \u2014NRbC(S)Rb, \u2014NRbC(O)ORb, \u2014NRbC(S)ORb, \u2014NRbC(O)NRcRc, \u2014NRbC(NRb)Rb or \u2014NRbC(NRb)NRcRc; and
wherein substituents used to substitute a specified group can be further substituted with one or more of the same or different substituent groups.
2. The compound of claim 1, wherein R21, R22, and R23 are all hydrogen.
3. The compound of claim 1, wherein R21 and R22 are all hydrogen.
4. The compound of claim 1 having a structural Formula (IIIb) or (IIIb’),
or a tautomer, andor salt thereof; wherein
Ll is alkylene or substituted alkylene;
L2 is \u2014NR34\u2014, \u2014O\u2014, \u2014S\u2014, \u2014NR34\u2014C(O)\u2014, \u2014C(O)\u2014NR34\u2014, \u2014O\u2014C(O)\u2014, \u2014C(O)\u2014O\u2014, \u2014NR34\u2014C(O)\u2014O\u2014, \u2014O\u2014C(O)\u2014NR34\u2014,\u2014NR34\u2014C(O)\u2014NR35\u2014, \u2014O\u2014C(O)\u2014O\u2014, -heterocyclylene-C(O)\u2014, or -(substituted heterocyclylene)-C(O)\u2014;
R33 is alkyl, substituted alkyl, carbocyclyl, substituted carbocyclyl; aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocyclyl, substituted heterocyclyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, or substituted heteroarylalkyl; and
R34 and R35 are independently hydrogen, alkyl, substituted alkyl, carbocyclyl, substituted carbocyclyl; aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocyclyl, substituted heterocyclyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, or substituted heteroarylalkyl.
5. The compound of claim 1 having a structural Formula (IIIc), or (IIIc’),
or a tautomer, andor salt, thereof; wherein
R33 is alkyl, substituted alkyl, carbocyclyl, substituted carbocyclyl; aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocyclyl, substituted heterocyclyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, or substituted heteroarylalkyl.
6. The compound of claim 5, wherein
R33 is alkyl, substituted alkyl, carbocyclyl, substituted carbocyclyl; heterocyclyl, substituted heterocyclyl, heteroalkyl, or substituted heteroalkyl.
7. The compound of claim 4, wherein
A is \u2014NH2;
B is hydrogen, or alkyl;
C is \u2014CO2R7;
L1 is alkylene or substituted alkylene;
L2 is \u2014NR34\u2014, \u2014O\u2014, \u2014NR34\u2014C(O)\u2014, \u2014C(O)\u2014NR34\u2014, \u2014O\u2014C(O)\u2014, \u2014C(O)\u2014O\u2014, -hetercyclylene-C(O)\u2014, or -(substituted hetercyclylene)-C(O)\u2014;
R33 is alkyl, substituted alkyl, carbocyclyl, substituted carbocyclyl; aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocyclyl, substituted heterocyclyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, or substituted heteroarylalkyl; and
R34 and R35 are independently hydrogen, alkyl, or substituted alkyl.
8. The compound of claim 5, wherein
A is \u2014NH2;
B is hydrogen, or alkyl;
C is \u2014CO2R7;
R33 is alkyl, substituted alkyl, carbocyclyl, substituted carbocyclyl; heterocyclyl, substituted heterocyclyl, heteroalkyl, or substituted heteroalkyl.
9. The compound of claim 1, which is selected from the group consisting of
10. The compound of claim 1, which is selected from the group consisting of
or a tautomer, andor salt, thereof.
The claims below are in addition to those above.
All refrences to claims which appear below refer to the numbering after this setence.
1. A monoclonal antibody that selectively binds to one of the following:
(a) hemoglobin variant HbD and glycated HbD, wherein the antibody binds to a HbD minimal epitope 121QFTPP125 (SEQ ID NO:8) or 119GKQFTPP125 (SEQ ID NO:46); or
(b) hemoglobin variant HbE and glycated HbE, wherein the antibody binds to a HbE minimal epitope 22EVGGK26 (SEQ ID NO:6) or 21DEVGGK26 (SEQ ID NO:7).
2. A method for individually detecting a plurality of hemoglobin-containing analytes comprising HbA1c and a hemoglobin variant, if present, in a single sample of blood cell lysate, said method comprising:
(a) incubating said sample with a population of beads in a common mixture, said population comprising a subpopulation to detect HbA1c and one or more further subpopulations, each of which detects a hemoglobin variant, wherein:
(i) each bead of said subpopulation to detect HbA1c has bonded thereto a classifier dye and each bead of said one or more subpopulations to detect a hemoglobin variant has bonded thereto a classifier dye where the classifier dyes are selected such that the classifier dyes, and thereby said subpopulations, are differentiable from each other by fluorescent emissions emitted by said classifier dyes upon excitation, and
(ii) each bead of said subpopulation to detect HbA1c further has bonded thereto an antibody having selective binding affinity towards HbA1c and each bead of said one or more subpopulations to detect a hemoglobin variant has bonded thereto a monoclonal antibody that has selective binding affinity towards the variant and the glycated form of the variant to cause each analyte to bind to the different bead subpopulation through the antibody bonded to said subpopulations, wherein:
one subpopulation of the one or more further subpopulations detects HbD, if present in the sample, and has bonded thereto a monoclonal antibody having selective binding affinity towards HbD and glycated HbD; or
one subpopulation of the one or more further subpopulations detects HbE, if present in the sample, and has bonded thereto a monoclonal antibody having selective binding affinity towards HbE and glycated HbE;
(b) with said analytes bound to the beads of said subpopulations, incubating said population with a labeled binding member that binds to all of said analytes, thereby labeling said analytes thus bound; and
(c) detecting the labeled binding member that binds to all of said analytes and detecting classifier dyes from the subpopulations of beads and differentiating the labels of the subpopulations by fluorescent emissions, thereby individually detecting HbA1c and if present, the hemoglobin variant.
3. The method of claim 2, wherein the population of beads further comprises a subpopulation that detects total hemoglobin, wherein each bead of the subpopulation to detect total hemoglobin has bonded thereto a classifier dye that is selected such that the classifier dye, and thereby said subpopulation, is differentiable from the other classifier dyes; and each member of said subpopulation that detects total hemoglobin having bonded thereto a monoclonal antibody having selective binding affinity towards all hemoglobin-containing analytes.
4. The method of claim 3, wherein said monoclonal antibody that has selective binding affinity towards all hemoglobin-containing analytes binds to a beta globin chain minimal epitope 9SAVTALWGKVNV20 (SEQ ID NO:15), 8KSAVTALWGKVNV20 (SEQ ID NO:16), or 11VTALW15 (SEQ ID NO:17).
5. The method of claim 2, wherein the antibody having selective binding affinity for HbA1c is a monoclonal antibody.
6. The method of claim 2, wherein one subpopulation of the one or more further subpopulations of (ii) detects HbD.
7. The method of claim 6, wherein the monoclonal antibody having selective affinity for HbD and glycated HbD is a monoclonal antibody that binds to a HbD minimal epitope 121QFTPP125 (SEQ ID NO:8) or 119GKQFTPP125 (SEQ ID NO:46).
8. The method of claim 2, wherein one subpopulation of the one or more further subpopulations of (ii) detects HbE.
9. The method of claim 8, wherein monoclonal antibody having selective affinity for HbE and glycated HbE is a monoclonal antibody that binds to a HbE minimal epitope 22EVGGK26 (SEQ ID NO:6) or 21DEVGGK26 (SEQ ID NO:7).
10. The method of claim 2, wherein the population comprises a subpopulation that detects HbD, if present in the sample, and a subpopulation that detects HbE, if present in the sample.
11. The method of claim 10, wherein the monoclonal antibody having selective binding affinity towards HbD and glycated HbD binds to a HbD minimal epitope 121QFTPP125 (SEQ ID NO:8) or 119GKQFTPP125 (SEQ ID NO:46); and the monoclonal antibody having selective affinity for HbE and glycated HbE binds to a HbE minimal epitope 22EVGGK26 (SEQ ID NO:6) or 21DEVGGK26 (SEQ ID NO:7).
12. The method of claim 2, wherein said method further comprises determining total hemoglobin in a non-immunoassay method.
13. The method of claim 2, wherein the blood cell lysate is a denatured blood cell lysate.
14. The method of claim 2, wherein the sample is from a diabetic patient.
15. A method for determining the proportion of HbA1c relative to total hemoglobin in a sample of blood cell lysate, adjusted for the possible presence in said lysate of a hemoglobin variant that interferes with the measurement of HbA1c, said method comprising:
(a) incubating said sample with a population of beads, said population consisting of a plurality of subpopulations in a common mixture, each bead of said population having bonded thereto one of a plurality of classifier dyes selected such that said classifier dyes, and thereby said subpopulations, are differentiable from each other by fluorescent emissions emitted by said classifier dyes upon excitation, each said subpopulation further having bonded thereto an immunological binding member having selective binding affinity toward one of a plurality of analytes, said plurality comprising HbA1c and said hemoglobin variant, to cause each of said analytes to bind to a different bead subpopulation through the immunological binding member bonded thereto;
(b) with said analytes bound to the beads of said subpopulations, incubating said population with a labeled binding member that binds to all of said analytes, thereby labeling said analytes so bound;
(c) with said bound analytes so labeled, detecting labels bound to said bound analytes while differentiating said labels so detected according to subpopulations by fluorescent emissions, thereby individually detecting concentrations of said analytes in said sample; and
(d) determining from said concentrations the proportion of HbA1c relative to total hemoglobin, adjusted for the concentration of said hemoglobin variant by an adjustment factor empirically derived from a predetermined relation between said hemoglobin variant concentration and the concentration of said HbA1c so detected.