1. A method for calibrating antimicrobic susceptibility testing data of microorganisms, wherein the method comprises the steps of
creating a histogram based on in vitro susceptibility test data against an antimicrobial agent for isolates from a microbial species, which microbial species isolates may contain unknown resistant strains against said antimicrobial agent wherein the number of isolates of microbial species or percentage of the total number of isolates of microbial species is on a y-axis of said histogram and registered response values against said antimicrobial agent on a x-axis of said histogram;
determining a position of a response peak of the histogram between a high response side and a low response side corresponding to isolates of microbial species susceptible to said antimicrobial agent;
calculating from said high response side of said histogram at least one statistical parameter; and
defining a limit for susceptibility interpretation and comparative analysis of antimicrobic resistance, which is based on said at least one statistical parameter; said limit separating susceptible strains from resistant strains against said antimicrobial agent.
2. Method as in claim 1, wherein said isolates arc clinical isolates.
3. Method as in claim 1, wherein said at least one statistical parameter is statistically obtained from a probability distribution calculated from said high response side of said histogram.
4. Method as in claim 1, wherein said microbial species is a bacterial species or a fungal species.
5. Method as in claim 1, wherein said antimicrobial agent is an antibiotic.
6. Method as in claim 1, wherein said antimicrobic susceptibility testing is a Minimal Inhibitory Concentration test, said registered response values being minimal inhibitory concentrations.
7. Method as in claim 1, wherein said antimicrobic susceptibility testing is a disk diffusion test, said registered response values being inhibition zone diameter values from paper disks impregnated with said antimicrobial agent.
8. Method as in claim 3, wherein said probability distribution is a Gaussian distribution.
9. Method as in claim 8, wherein said at least one statistical parameter is the mean, defining a position of the statistical response peak between a high statistical response distribution side and a low statistical response distribution side, andor standard deviation.
10. Method as in claim 9. wherein said limit is defined as three times said standard deviation below said mean.
11. Method as in claim 9, wherein said Gaussian distribution is a standard Gaussian distribution.
12. Method as in claim 9, wherein said Gaussian distribution is a normalized Gaussian distribution of a theoretical normal population.
13. Method as in claim 12, wherein said normalized Gaussian distribution is construed from said position of said statistical response peak, said high statistical response distribution side, and its mirror image.
14. Method as in claim 12, wherein a total number for said theoretical normal population is construed by a procedure comprising the steps of
calculating a slope for a line through said number of isolates of adjacent registered response values starting on said high response distribution side;
detecting a shift in slope direction, which represents the position of said statistical response peak, thereby defining said mean; and
calculating said total number as the doubled sum of half the number of isolates at said position plus the number of isolates having higher response values.
15. Method as in claim 13, wherein said normalized Gaussian distribution is construed graphically.
16. Method as in claim 14, wherein said normalized Gaussian distribution is determined by a procedure comprising the steps of
calculating for each registered response value a percentage value as percent of said total number;
calculating accumulated percentage values for each of said number of isolates or percent isolates on said statistical high response distribution side;
performing a linearized transformation of said accumulated percentage values against said statistical response values; and
calculating the equation constants for a linear relationship obtained from said transformation; thereby defining said normalized Gaussian distribution of said theoretical normal population.
17. Method as in claim 15, wherein said graphical construction is performed by means of an undirected graphical model or a directed graphical model.
18. Method as in claim 16, wherein said calculation of equation constants is performed by means of the least squares method.
19. Method as in claim 16, wherein said calculation of accumulated percentage values is performed in the direction 100% to 0%.
20. Method as in claim 16, wherein said transformation k a probit transformation.
21. Method as in claim 20, wherein said standard deviation is calculated from said probit transformation.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. At least one compound of the Formula I:
a pharmaceutically acceptable salt thereof, a solvate thereof, a chelate thereof, a non-covalent complex thereof, a prodrug thereof, and mixtures of any of the foregoing, wherein:
n is 1 to 6;
R1 is chosen from H, lower alkyl and substituted lower alkyl;
R2 is chosen from H, lower alkyl and substituted lower alkyl;
R3 and R4 are independently chosen from H, lower alkyl, substituted lower alkyl, lower haloalkyl, substituted lower haloalkyl, or R3 and R4 can join together to form a 3 to 6 membered ring or a substituted 3 to 6 membered ring;
R5 is chosen from OH, SH, NH2, lower alkyl, substituted lower alkyl, lower alkoxy, substituted lower alkoxy and sulfanyl;
R6 is chosen from H, OH, SH NH2, NHSO2R1 and sulfonyl;
each of R7, R8, R9 and R10 is independently chosen from H, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, NR3R4, C(O)OH, OR13, SR13, SO2R13, CN, NO2, halo, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heteroarylalkyl, substituted heteroarylalkyl, heterocycloalkyl, substituted heterocycloalkyl, alkylsilyl, substituted alkylsilyl, alkynylsilyl, substituted alkynylsilyl, alkoxy, substituted alkoxy, alkoxycarbonyl, substituted alkoxycarbonyl, and \u2014X\u2014R12, wherein:
R3 and R4 are defined above;
X is chosen from \u2014N(R11)\u2014Y\u2014 and \u2014Y\u2014N(R11)\u2014;
Y is chosen from C(O), SO2, alkylene, substituted alkylene, alkenylene, substituted alkenylene, alkynylene, and substituted alkynylene;
R11 is chosen from H, lower alkyl, and substituted lower alkyl,
R12 is chosen from H, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl; and
R13 is chosen from H, alkyl, substituted alkyl, alkenyl, substituted alkenyl; alkynyl, substituted alkynyl and NR3R4;
wherein optionally at least one of adjacent pairs R6 and R7, R7 and R8, R8 and R9, R9 and R10, and R10 and R1, join together to form a 4 to 7 membered ring or a substituted 4 to 7 membered ring.
2. The at least one compound according to claim 1, wherein R3 and R4 join together to form a 3 to 6 membered ring or a substituted 3 to 6 membered ring.
3. The at least one compound according to claim 2, wherein the 3 to 6 membered ring or the substituted 3 to 6 membered ring comprises at least one heteroatom.
4. The at least one compound according to claim 2, wherein the 3 to 6 membered ring or the substituted 3 to 6 membered ring comprises at least two heteroatoms.
5. The at least one compound according to claim 1, wherein R6 and R7 join together to form a 4 to 7 membered ring or a substituted 4 to 7 membered ring.
6. The at least one compound according to claim 5, wherein the 4 to 7 membered ring or the substituted 4 to 7 membered ring comprises at least one heteroatom.
7. The at least one compound according to claim 5 wherein the 4 to 7 membered ring or the substituted 4 to 7 membered ring comprises at least two heteroatoms.
8. The at least one compound according to claim 5 wherein the 4 to 7 membered ring or the substituted 4 to 7 membered ring comprises at least three heteroatoms.
9. The at least one compound according to claim 1, wherein at least one of R7, R8, R9 and R10 is independently chosen from halo and a moiety substituted with at least one halo.
10. The at least one compound according to claim 1, wherein at least one of R7, R8, R9 and R10 is independently chosen from alkoxy or substituted alkoxy.
11. The at least one compound according to claim 1, wherein at least one of R7, R8, R9 and R10 is independently chosen from alkylsilyl, substituted alkylsilyl, alkynylsilyl, and substituted alkynylsilyl.
12. The at least one compound according to claim 1, wherein at least one of R7, R8, R9 and R10 is independently chosen from aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycloalkyl, and substituted heterocycloalkyl.
13. The at least one compound according to claim 1, wherein at least one of R7, R8, R9 and R10 is independently chosen from H, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, and substituted alkynyl.
14. The at least one compound according to claim 1 having the structure of any one of Compounds 1-175 set forth in Table 1.
15. A pharmaceutical composition comprising at least one pharmaceutically acceptable excipient, and a therapeutically effective amount of at least one compound according to claim 1.
16. The pharmaceutical composition of claim 15, wherein the at least one compound is present in an amount effective for the treatment of at least one disease chosen from ischemia, anemia, wound healing, auto-transplantation, allo-transplantation, xeno-transplantation, systemic high blood pressure, thalassemia, diabetes, cancer and an inflammatory disorder.
17. A method of increasing HIF levels or activity in a subject administering to the subject at least one compound according to claim 1.
18. A method of treating a condition where it is desired to modulate HIF activity comprising administering to a subject at least one compound according to claim 1.
19. A method according to claim 18, wherein said condition is chosen from at least one of ischemia, anemia, wound healing, auto-transplantation, allo-transplantation, xeno-transplantation, systemic high blood pressure, thalassemia, diabetes, cancer and an inflammatory disorder.
20. A method of treating a hypoxic or ischemic related disorder in a subject comprising administering to a subject at least one compound according to claim 1.
21. A method of modulating the amount of HIF in a cell comprising contacting the cell with at least one compound according to claim 1.
22. A method of increasing the amount of hemoglobin F in a subject comprising administering to the subject at least one compound according to claim 1.
23. A method of modulating angiogenesis in a subject comprising administering to the subject at least one compound according to claim 1.
24. A method of treating at least one disease in a patient in need of such treatment comprising administering to the patient a therapeutically effective amount of at least one compound according to claim 1.
25. The method according to claim 24, wherein the at least one disease is chosen from ischemia, anemia, wound healing, auto-transplantation, allo-transplantation, xeno-transplantation, systemic high blood pressure, thalassemia, diabetes, cancer and an inflammatory disorder.
26. A method of inhibiting HIF hydroxylation in a subject comprising administering to the subject at least one compound according to claim 1.
27. The at least one compound according to claim 1, wherein the HIF PHD inhibitory activity IC50 value is 40 \u03bcM or less.
28. The at least one compound according to claim 1, wherein the HIF PHD inhibitory activity IC50 value is 1.0 \u03bcM or less.
29. An assay for the detection of HIF1\u03b1 hydroxyproline residues comprising:
a) incubating a fluorochrome-labeled HIF1\u03b1 polypeptide or fragment thereof with a VCB complex labeled with a rare earth element;
b) detecting the binding of the VCB complex to HIF1\u03b1 by homogeneous time-resolved FRET.
30. The assay according to claim 29, wherein the fluorochrome is allophycocyanin.
31. The assay according to claim 29, wherein the rare earth element is europium.
32. An assay for the detection of HIF1\u03b1 hydroxyproline residues comprising:
a) incubating a HIF1\u03b1 polypeptide or fragment thereof with a VCB complex labeled with ruthenium;
b) detecting the binding of the VCB complex to HIF1\u03b1 by electrochemiluminescence.
33. The assay according to claim 32, wherein the HIF1\u03b1 polypeptide or fragment thereof is bound to a solid support.