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C.: \"On the generalised distance in statistics\", PROCEEDINGS OF THE NATIONAL INSTITUTE OF SCIENCES OF INDIA, vol. 2, no. 1, 1936, pages 49 - 55, Retrieved from the Internet(a) incubating cells infected with mycobacteria with a test compound;
(b) staining the cells with (a) detectable marker probe(s) or introducing into the cell a detectable marker probe;
(c) obtaining a multi-parametric phenotypic profile of host cell parameters and bacterial parameters; and
(d) clustering the multi-parametric phenotypic profile by using a modified Gabriel Neighbourhood and density estimate algorithm, which comprises:"],"number":6,"annotation":false,"title":false,"claim":true},{"lines":["The method according to claim 6, wherein the cells are infected with mycobacteria that express a fluorescent protein marker."],"number":7,"annotation":false,"title":false,"claim":true},{"lines":["The method according to claim 6 or 7, wherein the bacterial parameters are selected from the group consisting of parameters related to the number of bacteria, parameters related to signal intensity of the bacteria and parameters related to the shape of the bacteria."],"number":8,"annotation":false,"title":false,"claim":true},{"lines":["The method according to any one of claims 6 to 8, wherein the host cell parameters are selected from the group consisting of parameters related to the morphology of the cell and parameters indicative of toxicity."],"number":9,"annotation":false,"title":false,"claim":true},{"lines":["The method according to any one of claims 6 to 9, wherein the bacterial parameters comprise at least(i) computing multi-dimensional Gabriel neighbourhood graph over dataset, where nodes correspond to phenotypic profiles and edges determine the neighbourhood of nodes;(ii) filtering spurious as well as 'long-range' edges from the Gabriel graph;(iii) computing the density estimate for each node of the Gabriel graph using a kernel function, wherein the kernel function is chosen according to the selected multi-parametric profile dissimilarity measure;(iv) converting the modified Gabriel graph into weighted directed acyclic graph (WDAG) by assigning directionality to edges such that their point from nodes with lower density estimate to nodes with higher density estimate and edge weights are set proportional to the distances (dissimilarity) between neighbouring nodes, or, in the ambiguous case of equal density estimate of two neighbour nodes, one node is randomly chosen and a small arbitrary value (0.0000001) is added to its density estimate;(v) marking the nodes of the graph with no out-coming edges, ie. nodes having local maxima of density, as cluster centres; and(vi) assigning the remaining nodes to the cluster centers based on the shortest weighted path in the directed neighbourhood graph;wherein an alteration in bacterial parameters and a lack or substantial lack of alteration of host cell parameters in the presence of the test compound is indicative of a compound suitable as a lead compound for the treatment of tuberculosis.
(i) number of bacteria per cell (mean);
(ii) total number of bacteria (per image);
(iii) integral vesicle intensity of bacteria within infected cell;
(iv) mean intensity of bacteria (non-weighted median);
(v) mean area of bacteria (non-weighted median); and
(vi) number of infected cells;and wherein the host cell parameters comprise at least
(a) number of cells."],"number":10,"annotation":false,"title":false,"claim":true},{"lines":["The method of any one of claims 6 to 10, further comprising optimizing the pharmacological properties of the test compound identified as lead compound."],"number":11,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 11, wherein the optimization comprises modifying the test compound identified as lead compound to achieve:
i) modified spectrum of activity, organ specificity, and/or
ii) improved potency, and/or
iii) decreased toxicity (improved therapeutic index), and/or
iv) decreased side effects, and/or
v) modified onset of therapeutic action, duration of effect, and/or
vi) modified pharmacokinetic parameters (resorption, distribution, metabolism and excretion), and/or
vii) modified physico-chemical parameters (solubility, hygroscopicity, colour, taste, odour, stability, state), and/or
viii) improved general specificity, organ/tissue specificity, and/or
ix) optimised application form and route by"],"number":12,"annotation":false,"title":false,"claim":true},{"lines":["A method of identifying the cellular pathway targeted by a test compound, the method comprising(a) esterification of carboxyl groups, or(b) esterification of hydroxyl groups with carboxylic acids, or(c) esterification of hydroxyl groups to, e.g. phosphates, pyrophosphates or sulfates or hemi-succinates, or(d) formation of pharmaceutically acceptable salts, or(e) formation of pharmaceutically acceptable complexes, or(f) synthesis of pharmacologically active polymers, or(g) introduction of hydrophilic moieties, or(h) introduction/exchange of substituents on aromates or side chains, change of substituent pattern, or(i) modification by introduction of isosteric or bioisosteric moieties, or(j) synthesis of homologous compounds, or(k) introduction of branched side chains, or(l) conversion of alkyl substituents to cyclic analogues, or(m) derivatisation of hydroxyl groups to ketales, acetales, or(n) N-acetylation to amides, phenylcarbamates, or(o) synthesis of Mannich bases, imines, or(p) transformation of ketones or aldehydes to Schiffs bases, oximes, acetales, ketales, enolesters, oxazolidines, thiazolidinesor combinations thereof.
(a) treating a cell with the test compound and obtaining a multi-parametric phenotypic profile of the cell after said treatment;
(b) providing a phenotypic profile obtained for the same cell type by introducing directed genetic perturbations to target genes of said cell;
(c) comparing the multi-parametric phenotypic profile obtained in (a) with the phenotypic profile provided in (b) and identifying the genes whose perturbation leads to an identical or substantially identical phenotypic profile as the test compound; and
(d) identifying the cellular pathways involving the genes identified in (c)."],"number":13,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 13, wherein the directed genetic perturbations to target genes of the cell are introduced by use of RNAi."],"number":14,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 14, wherein the RNAi-mediated genetic perturbation is a genome-wide RNAi screen."],"number":15,"annotation":false,"title":false,"claim":true}]}},"filters":{"npl":[],"notNpl":[],"applicant":[],"notApplicant":[],"inventor":[],"notInventor":[],"owner":[],"notOwner":[],"tags":[],"dates":[],"types":[],"notTypes":[],"j":[],"notJ":[],"fj":[],"notFj":[],"classIpcr":[],"notClassIpcr":[],"classNat":[],"notClassNat":[],"classCpc":[],"notClassCpc":[],"so":[],"notSo":[],"sat":[]},"sequenceFilters":{"s":"SEQIDNO","d":"ASCENDING","p":0,"n":10,"sp":[],"si":[],"len":[],"t":[],"loc":[]}}