RELATING TO DRUG METABOLISM

PRIORITY

[0001] This application claims priority to U.S. Provisional Application No. 62/153,755 entitled "System and Method for Processing Genotype Information Relating to Drug Metabolism" by Brian Meshkin filed on April 28, 2015, which is incorporated herein by reference in its entirety.

BACKGROUND

[0002] In nature, organisms of the same species usually differ from each other in various aspects such as in their appearance or in one or more aspects of their biology. The differences are often based on genetic distinctions, some of which are called polymorphisms. Polymorphisms are often observed at the level of the whole individual (i.e., phenotype polymorphism), in variant forms of proteins and blood group substances (i.e., biochemical polymorphism), in morphological features of chromosomes (i.e., chromosomal polymorphism), and at the level of DNA in differences of nucleotides and/or nucleotide sequences (i.e., genetic polymorphism).

[0003] Examples of genetic polymorphisms include alleles and haplotypes. An allele is an alternative form of a gene, such as one member of a pair, that is located at a specific position on a chromosome and are known as single nucleotide polymorphisms (SNPs). A haplotype is a combination of alleles, or a combination of SNPs on the same chromosome. An example of a genetic polymorphism is an occurrence of one or more genetically alternative phenotypes in a subject due to the presence or absence of an allele or haplotype. [0004] Genetic polymorphisms can play a role in determining differences in an individual's response to a species of drug, a drug dosage or a therapy including one drug or a combination of drugs. Pharmacogenetics and pharmacogenomics are multidisciplinary research efforts to study the relationships among genotypes, gene expression profiles, and phenotypes, as often expressed through the variability between individuals in response to the drugs taken. Since the initial sequencing of the human genome, more than a million S Ps have been identified. Some of these SNPs have been used to predict clinical predispositions or responses based upon data gathered from pharmacogenomic studies.

[0005] A patient's genotype information is often utilized to help a prescriber decide between medications based on information associated with a patient's genetic profile (i.e., genotype information). There is a desire to utilize a patient's genotype information in determining the patient's predisposition to drug metabolism of individual drugs and drug panels. There is also a desire for methods for predicting and/or diagnosing individuals exhibiting irregular drug metabolism of individual drugs and drug panels. Furthermore, there is also a desire to determine genetic information, such as polymorphisms, which may be utilized for predicting variations in predisposition to metabolism of individual drugs and drug panels. There is also a desire to implement systems processing and distribing the detected genetic information in a systematic way. Such genetic information would be useful in providing prognostic information about treatment options for a patient based on the patient's metabolism of individual drugs and drug panels.

[0006] Although it is known generally that drug metabolism may be associated with genetics - a factor not routinely considered, there is no rigourous methodology to systematically provide doctors with an ability to identify patients exhibiting irregular drug metabolism of individual drugs and drug panels. Such systems and methods would be beneficial to provide information that improves accuracy in identifying patients at risk for irregular drug metabolism.

[0007] Given the foregoing, and to address the above-described limitations, systems and methods are desired for identifying, estimating and/or determining a potential for success of an individual patient's clinical outcome in response to being treated with one or more drugs based on the patient's metabolism.

SUMMARY

[0008] This summary is provided to introduce a selection of concepts that are further described in the Detailed Description below. The genes, polymorphisms, sequences and sequence identifiers (i.e., SEQ IDs or SEQ ID Numbers) listed or referenced herein are also described in greater detail below in the Detailed Description. This summary is not intended to identify key or essential features of the claimed subject matter. Also, this summary is not intended as an aid in determining the scope of the claimed subject matter.

[0009] The present invention meets the above-identified needs by providing systems, methods and computer readable mediums (CRMs) for preparing and utilizing prognostic information associated with a predisposition to risk for irregular drug metabolism in a patient. The prognostic information is derived from genotype information about a patient's gene profile. The genotype information may be obtained by, inter alia, assaying a sample of genetic material associated with a patient.

[0010] The systems, methods and CRMs, according to the principles of the invention, can be utilized to determine prognostic information associated with drug metabolism in a patient. The prognostic information may be used for addressing prescription needs directed to caring for an individual patient. It may also be utilized in managing large healthcare entities, such as insurance providers, utilizing comprehensive business intelligence systems. These and other objects are accomplished by systems, methods and CRMs directed to preparing and utilizing prognostic information associated with drug metabolism in a patient, in accordance with the principles of the invention.

[0011] According to a first principal of the invention, there is a method. The method may include facilitating a processing of and/or processing (1) data and/or (2) information and/or (3) at least one signal, the (1) data and/or (2) information and/or (3) at least one signal based, at least in part, on any combination of at least part of the following: determining patient information, including DNA information, associated with a human subject; determining from the DNA information whether a subject genotype of the human subject includes one or more alleles or haplotypes in at least one gene by detecting, utilizing a detection technology and the DNA information, a presence or absence of the one or more alleles or haplotypes in the one or more genes of the subject genotype, wherein the at least one gene is selected from the gene group: CYP2C8, CYP2C9, CYP2C19, CYP3A4, CYP3A5, CYP2D6, CYP1A2, VKORC1, UGT2B7, CYP2B6 and CYP2E1; determining at least one allele score associated with the alleles or haplotypes in the one or more genes based on the detected presence or absence of the one or more alleles or haplotypes in the one or more genes; determining at least one grade associated with one or more genes of the at least one gene based on the determined at least one allele score; and determining a drug metabolism response by the human subject associated with at least one drug based on the determined at least one grade associated with the one or more genes of the at least one gene. [0012] The method may also include wherein the (1) data and/or (2) information and/or

(3) at least one signal are further based, at least in part, on any combination of the following: determining at least one drug dosage recommendation based on the determined drug metabolism response associated with the at least one drug, wherein the method for determining the drug metabolism response associated with the human subject, is an ex vivo method; determining a comparing of a region, including the one or more alleles or haplotypes, of the subject genotype with a corresponding region of a predetermined reference genotype, wherein characteristics of the corresponding region of the reference genotype are based upon a predetermined population norm; determining prognostic information associated with the human subject based on the determined drug metabolism response; and determining a therapy for the human subject based on the determined prognostic information associated with the human subject, wherein the at least one gene includes at least any number from two to eleven genes selected from the gene group.

[0013] According to a second principal of the invention, there is an apparatus. The apparatus may include any combination of at least one processor; and at least one memory including computer program code for one or more programs, the at least one memory and the computer program code configured to, with the at least one processor, cause the apparatus to perform at least the following, determine patient information, including DNA information, associated with a human subject; determine from the DNA information whether a subject genotype of the human subject includes one or more alleles or haplotypes in at least one gene by detecting, utilizing a detection technology and the DNA information, a presence or absence of the one or more alleles or haplotypes in the one or more genes of the subject genotype, wherein the at least one gene is selected from the gene group: CYP2C8, CYP2C9, CYP2C19, CYP3A4, CYP3A5, CYP2D6, CYP1A2, VKORCl, UGT2B7, CYP2B6 and CYP2E1; determine at least one allele score associated with the alleles or haplotypes in the one or more genes based on the detected presence or absence of the one or more alleles or haplotypes in the one or more genes; determine at least one grade associated with one or more genes of the at least one gene based on the determined at least one allele score; and determine a drug metabolism response by the human subject associated with at least one drug based on the determined at least one grade associated with the one or more genes of the at least one gene.

[0014] According to a third principal of the invention, there is a non-transitory computer readable medium. The medium may store any combination of computer readable instructions that when executed by at least one processor perform a method, the method comprising facilitating a processing of and/or processing (1) data and/or (2) information and/or (3) at least one signal, the (1) data and/or (2) information and/or (3) at least one signal based, at least in part, on any combination of the following: determining patient information, including DNA information, associated with a human subject; determining from the DNA information whether a subject genotype of the human subject includes one or more alleles or haplotypes in at least one gene by detecting, utilizing a detection technology and the DNA information, a presence or absence of the one or more alleles or haplotypes in the one or more genes of the subject genotype, wherein the at least one gene is selected from the gene group: CYP2C8, CYP2C9, CYP2C19, CYP3A4, CYP3A5, CYP2D6, CYP1A2, VKORC1, UGT2B7, CYP2B6 and CYP2E1; determining at least one allele score associated with the alleles or haplotypes in the one or more genes based on the detected presence or absence of the one or more alleles or haplotypes in the one or more genes; determining at least one grade associated with one or more genes of the at least one gene based on the determined at least one allele score; and determining a drug metabolism response by the human subject associated with at least one drug based on the determined at least one grade associated with the one or more genes of the at least one gene.

[0015] The above summary is not intended to describe each embodiment or every implementation of the present invention. Further features, their nature and various advantages are made more apparent from the accompanying drawings and the following examples and embodiments.

BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Features and advantages of the present invention become more apparent from the detailed description, set forth below, when taken in conjunction with the drawings. In the drawings, like reference numbers indicate identical or functionally similar elements. Additionally, a left-most digit of a reference number identifies a drawing in which the reference number first appears. In addition, it should be understood that the drawings in the figures which highlight an aspect, methodology, functionality and/or advantage of the present invention, are presented for example purposes only. The present invention is sufficiently flexible such that it may be implemented in ways other than shown in the accompanying figures.

[0017] FIG. 1 is a block diagram illustrating an assay system which may be utilized for preparing genotype information from a sample of genetic material, according to an example;

[0018] FIG. 2 is a block diagram illustrating a prognostic information system which may be utilized for preparing and/or utilizing prognostic information utilizing the genotype information prepared using the assay system of FIG. 1, according to an example;

[0019] FIG. 3 is a flow diagram illustrating a prognostic information process for identifying a risk to a patient utilizing the assay system of FIG. 1 and the prognostic information system of FIG. 2, according to an example; and

[0020] FIG. 4 is a block diagram illustrating a computer system providing a platform for the assay system of FIG. 1 or the prognostic information system of FIG. 2, according to various examples.

DETAILED DESCRIPTION

[0021] The present invention is useful for preparing and/or utilizing prognostic information about a patient. The prognostic information may be utilized to determine an appropriate therapy for the patient based on their genotype and phenotype information to identify their genetic predisposition to drug metabolim risk. The genetic predisposition may be associated with the selection of an individual drug or a combination of drugs, a dosage of the medication(s) and the utilization of the medication(s) in a regimen for treating the patient's medical condition.

[0022] The prognostic information may also be utilized for determining dose adjustments that may help a prescriber understand why a patient is or is not responding to a medication dosage, such as an "average" dose. The prognostic information may also be utilized by a prescriber to decide between medications based on the patient's genetic predisposition to drug metabolism risk to various drugs.

[0023] The prognostic information may also be utilized for predicting and/or diagnosing individuals exhibiting a regular or irregular predisposition to drug metabolism risk. Such genetic information can be very useful in providing prognostic information about treatment options for a patient. The patient may be associated with a medical condition. The patient may also have already been prescribed a medication for treating the medical condition. The present invention has been found to be advantageous for determining a treatment for a patient who may have a regular or irregular predisposition to drug metabolism risk. While the present invention is not necessarily limited to such applications, various aspects of the invention may be appreciated through a discussion of the various examples in this context, as illustrated through the examples below.

[0024] For simplicity and illustrative purposes, the present invention is described by referring mainly to embodiments, principles and examples thereof. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the examples. It is readily apparent however, that the embodiments may be practiced without limitation to these specific details. In other instances, some embodiments have not been described in detail so as not to unnecessarily obscure the description. Furthermore, different embodiments are described below. The embodiments may be used or performed together in different combinations.

[0025] The operation and effects of certain embodiments can be more fully appreciated from the examples described below. The embodiments on which these examples are based are representative only. The selection of embodiments is to illustrate the principles of the invention and does not indicate that variables, functions, conditions, techniques, configurations and designs, etc., which are not described in the examples are not suitable for use, or that subject matter not described in the examples is excluded from the scope of the appended claims and their equivalents. The significance of the examples can be better understood by comparing the results obtained therefrom with potential results which can be obtained from tests or trials that may be or may have been designed to serve as controlled experiments and provide a basis for comparison. [0026] Before the systems and methods are described, it is understood that the invention is not limited to the particular methodologies, protocols, systems, platforms, assays, and the like which are described, as these may vary. It is also to be understood that the terminology used herein is intended to describe particular embodiments of the present invention, and is in no way intended to limit the scope of the present invention as set forth in the appended claims and their equivalents.

[0027] Throughout this disclosure, various publications, such as patents and published patent specifications, are referenced by an identifying citation. The disclosures of these publications are hereby incorporated by reference in their entirety into the present disclosure in order to more fully describe the state of the art to which the invention pertains.

[0028] The practice of the present invention employs, unless otherwise indicated, conventional techniques of molecular biology, microbiology, cell biology, biochemistry and immunology, which are within the skill of the art. Such techniques are explained fully in the literature for example in the following publications. See, e.g., Sambrook and Russell eds. MOLECULAR CLONING: A LABORATORY MANUAL, 3rd edition (2001); the series CURRENT PROTOCOLS IN MOLECULAR BIOLOGY (F. M. Ausubel et al. eds. (2007)); the series METHODS IN ENZYMOLOGY (Academic Press, Inc., N Y ); PCR 1 : A PRACTICAL APPROACH (M. MacPherson et al. IRL Press at Oxford University Press (1991)); PCR 2: A PRACTICAL APPROACH (M. J. MacPherson, B. D. Hames and G. R. Taylor eds. (1995)); ANTIBODIES, A LABORATORY MANUAL (Harlow and Lane eds. (1999)); CULTURE OF ANFMAL CELLS: A MANUAL OF BASIC TECHNIQUE (R. I. Freshney 5th edition (2005)); OLIGONUCLEOTIDE SYNTHESIS (M. J. Gait ed. (1984)); Mullis et al., U.S. Pat. No. 4,683,195; NUCLEIC ACID HYBRIDIZATION (B. D. Hames & S. J. Higgins eds. (1984)); NUCLEIC ACID HYBRIDIZATION (M. L. M. Anderson (1999)); TRANSCRIPTION AND TRANSLATION (B. D. Hames & S. J. Higgins eds. (1984)); IMMOBILIZED CELLS AND ENZYMES (IRL Press (1986)); B. Perbal, A PRACTICAL GUIDE TO MOLECULAR CLONING (1984); GENE TRANSFER VECTORS FOR MAMMALIAN CELLS (J. H. Miller and M. P. Calos eds. (1987) Cold Spring Harbor Laboratory); GENE TRANSFER AND EXPRESSION IN MAMMALIAN CELLS (S. C. Makrides ed. (2003)) IMMUNOCHEMICAL METHODS IN CELL AND MOLECULAR BIOLOGY (Mayer and Walker, eds., Academic Press, London (1987)); WEIR'S HANDBOOK OF EXPERFMENTAL FMMUNOLOGY (L. A. Herzenberg et al. eds (1996)); MANIPULATING THE MOUSE EMBRYO: A LABORATORY MANUAL 3rd edition (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N. Y. (2002)). DEFINITIONS.

[0029] As used herein, certain terms have the following defined meanings. As used herein, the singular form "a," "a and "the" includes the singular and plural references unless the context clearly dictates otherwise. For example, the term "a cell" includes a single cell and a plurality of cells, including mixtures thereof.

[0030] As used herein, the terms "based on," "comprises," "comprising," "includes,"

"including," "has," "having" or any other variation thereof, are intended to cover a non-exclusive inclusion. For example, a system, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such system, process, method, article, or apparatus. Further, unless expressly stated to the contrary, "or" refers to an inclusive or and not to an exclusive or. For example, a condition A or B is satisfied by any one of the following: A is true (or present) and B is false (or not present), A is false (or not present) and B is true (or present), and both A and B is true (or present).

[0031] All numerical designations, e.g., pH, temperature, time, concentration, and molecular weight, including ranges, are approximations which may be varied (+) or (-) by minor increments, such as, of 0.1. It is to be understood, although not always explicitly stated, that all numerical designations are preceded by the term "about". The term "about" also includes the exact value "X" in addition to minor increments of "X" such as "X+0.1" or "X-0.1." It also is to be understood, although not always explicitly stated, that the reagents described herein are merely exemplary and that equivalents of such are known to those of ordinary skill in the art.

[0032] The term "allele" which is used interchangeably herein with the term "allelic variant" refers to alternative forms of a gene or any portions thereof. Alleles may occupy the same locus or position on homologous chromosomes. When a subject has two identical alleles of a gene, the subject is said to be homozygous for the gene or allele. When a subject has two different alleles of a gene, the subject is said to be heterozygous for the gene or allele. Alleles of a specific gene can differ from each other in a single nucleotide, or several nucleotides, and can include substitutions, deletions and insertions of nucleotides. An allele of a gene can also be an ancestral form of a gene or a form of a gene containing a mutation.

[0033] The term "haplotype" refers to a combination of alleles on a chromosome or a combination of SNPs within an allele on one chromosome. The alleles or SNPs may or may not be at adjacent locations (loci) on a chromosome. A haplotype may be at one locus, at several loci or an entire chromosome.

[0034] The term "ancestral," when applied to describe an allele in a human, refers to an allele of a gene that is the same or nearest to a corresponding allele appearing in the corresponding gene of the chimpanzee genome. Often, but not always, a human ancestral allele is the most prevalent human allelic variant appearing in nature - i.e., the allele with the highest gene frequency in a population of the human species.

[0035] The term "wild-type," when applied to describe an allele, refers to an allele of a gene which, when it is present in two copies in a subject, results in a wild-type phenotype. There can be several different wild-type alleles of a specific gene. Also, nucleotide changes in a gene may not affect the phenotype of a subject having two copies of the gene with the nucleotide changes.

[0036] The term "polymorphism" refers to the coexistence of more than one form of a gene or portion thereof. A portion of a gene of which there are at least two different forms, i.e., two different nucleotide sequences, is referred to as a "polymorphic region of a gene." A polymorphic region may include, for example, a single nucleotide polymorphism (S P), the identity of which differs in the different alleles by a single nucleotide at a locus in the polymorphic region of the gene. In another example, a polymorphic region may include a deletion or substitution of one or more nucleotides at a locus in the polymorphic region of the gene.

[0037] The expression "amplification of polynucleotides" includes methods such as PCR, ligation amplification (or ligase chain reaction, LCR) and other amplification methods. These methods are known and widely practiced in the art. See, e.g., U.S. Pat. Nos. 4,683,195 and 4,683,202 and Innis et al., 1990 (for PCR); and Wu et al. (1989) Genomics 4:560-569 (for LCR). In general, a PCR procedure is a method of gene amplification which is comprised of (i) sequence-specific hybridization of primers to specific genes within a DNA sample (or library), (ii) subsequent amplification involving multiple rounds of annealing, elongation, and denaturation using a DNA polymerase, and (iii) screening the PCR products for a band of the correct size. The primers used are oligonucleotides of sufficient length and appropriate sequence to provide initiation of polymerization, i.e., each primer is specifically designed to be complementary to each strand of the genomic locus to be amplified.

[0038] Reagents and hardware for conducting PCR are commercially available. Primers useful to amplify sequences from a particular gene region are preferably complementary to, and hybridize specifically to sequences in the target region or in its flanking regions. Nucleic acid sequences generated by amplification may be sequenced directly. Alternatively, the amplified sequence(s) may be cloned prior to sequence analysis. Methods for direct cloning and sequence analysis of enzymatically amplified genomic segments are known in the art.

[0039] The term "encode," as it is applied to polynucleotides, refers to a polynucleotide which is said to "encode" a polypeptide. The polynucleotide is transcribed to produce mRNA, which is then translated into the polypeptide and/or a fragment thereof by cell machinery. An antisense strand is the complement of such a polynucleotide, and the encoding sequence can be deduced therefrom.

[0040] As used herein, the term "gene" or "recombinant gene" refers to a nucleic acid molecule comprising an open reading frame and including at least one exon and optionally an intron sequence. The term "intron" refers to a DNA sequence present in a given gene which is spliced out during mRNA maturation.

[0041] "Homology" or "identity" or "similarity" refers to sequence similarity between two peptides or between two nucleic acid molecules. Homology can be determined by comparing a position in each sequence which may be aligned for purposes of comparison. When a position in the compared sequence is occupied by the same base or amino acid, then the molecules are homologous at that position. A degree of homology between sequences is a function of the number of matching or homologous positions shared by the sequences. A "related" or "homologous" sequence shares identity with a comparative sequence, such as 100%, at least 99%, at least 95%, at least 90%, at least 80%, at least 70%, at least 60%, at least 50%, at least 40%), at least 30%, at least 20%, or at least 10%. An "unrelated" or "non-homologous" sequence shares less identity with a comparative sequence, such as less than 95%, less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, or less than 10%.

[0042] The term "a homolog of a nucleic acid" refers to a nucleic acid having a nucleotide sequence having a certain degree of homology with the nucleotide sequence of the nucleic acid or complement thereof. A homolog of a double stranded nucleic acid is intended to include nucleic acids having a nucleotide sequence which has a certain degree of homology with or with the complement thereof. In one aspect, homologs of nucleic acids are capable of hybridizing to the nucleic acid or complement thereof.

[0043] The term "isolated" as used herein with respect to nucleic acids, such as DNA or

RNA, refers to molecules separated from other DNAs or RNAs, respectively, which are present in a natural source of a macromolecule. The term isolated as used herein also refers to a nucleic acid or peptide that is substantially free of cellular material, viral material, or culture medium when produced by recombinant DNA techniques, or chemical precursors or other chemicals when chemically synthesized. Moreover, an "isolated nucleic acid" is meant to include nucleic acid fragments which are not naturally occurring as fragments and would not be found in the natural state. The term "isolated" is also used herein to refer to polypeptides which are isolated from other cellular proteins and is meant to encompass both purified and recombinant polypeptides. [0044] As used herein, the term "nucleic acid" refers to polynucleotides such as deoxyribonucleic acid (DNA), and, where appropriate, ribonucleic acid (RNA). The term "nucleic acid" should also be understood to include, as equivalents, derivatives, variants and analogs of either RNA or DNA made from nucleotide analogs, and, as applicable to the embodiment being described, single (sense or antisense) and double-stranded polynucleotides.

[0045] Deoxyribonucleotides include deoxyadenosine, deoxycytidine, deoxyguanosine, and deoxythymidine. For purposes of clarity, when referring herein to a nucleotide of a nucleic acid, which can be DNA or RNA, the terms "adenosine," "cytidine," "guanosine," and "thymidine" are used. It is understood that if the nucleic acid is RNA, it includes nucleotide(s) having a uracil base that is uridine.

[0046] The terms "oligonucleotide" or "polynucleotide," or "portion," or "segment" thereof refer to a stretch of polynucleotide residues which may be long enough to use in PCR or various hybridization procedures to identify or amplify identical or related parts of mRNA or DNA molecules. The polynucleotide compositions described herein may include RNA, cDNA, genomic DNA, synthetic forms, and mixed polymers, both sense and antisense strands, and may be chemically or biochemically modified or may contain non-natural or derivatized nucleotide bases, as will be readily appreciated by those skilled in the art. Such modifications can include, for example, labels, methylation, substitution of one or more of the naturally occurring nucleotides with an analog, internucleotide modifications such as uncharged linkages (e.g., methyl phosphonates, phosphotriesters, phosphoamidates, carbamates, etc.), charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.), pendent moieties (e.g., polypeptides), intercalators (e.g., acridine, psoralen, etc.), chelators, alkylators, and modified linkages (e.g., alpha anomeric nucleic acids, etc.). This may also include synthetic molecules that mimic polynucleotides in their ability to bind to a designated sequence via hydrogen bonding and other chemical interactions. Such molecules are known in the art and include, for example, those in which peptide linkages substitute for phosphate linkages in the backbone of the molecule.

[0047] The phrase "genetic profile" is used interchangeably with "genotype information" and refers to part or all of an identified genotype of a subject and may include one or more polymorphisms in one or more genes of interest. A genetic profile may not be limited to specific genes and polymorphisms described herein, and can include any number of other polymorphisms, gene expression levels, polypeptide sequences, or other genetic markers that are associated with a subject or patient.

[0048] The term "patient" refers to an individual waiting for or under medical care and treatment, such as a treatment for medical condition. While the disclosed methods are designed for human patients, such methods are applicable to any suitable individual, which includes, but is not limited to, a mammal, such as a mouse, rat, rabbit, hamster, guinea pig, cat, dog, goat, cow, horse, pig, and simian. Human patients include male and female patients of any ethnicity. The term "treating" as used herein is intended to encompass curing as well as ameliorating at least one symptom of a condition or disease.

[0049] The nucleic acid codes utilized herein include: A for Adenine, C for Cytosine, G for Guanine, T for Thymine, and U for Uracil.

[0050] As used herein, the terms "drug," "medication," and "therapeutic compound" or

"compound" are used interchangeably and refer to any chemical entity, pharmaceutical, drug, biological, and the like that can be used to treat or prevent a disease, illness, condition, or disorder of bodily function. A drug may comprise both known and potentially therapeutic compounds. A drug may be determined to be therapeutic by screening using the screening known to those having ordinary skill in the art. A "known therapeutic compound" or "medication" refers to a therapeutic compound that has been shown (e.g., through animal trials or prior experience with administration to humans) to be effective in such treatment. Examples of drugs include, but are not limited to peptides, polypeptides, synthetic organic molecules, naturally occurring organic molecules, nucleic acid molecules, and combinations thereof.

[0051] The biological basis for an outcome in a specific patient following a treatment with a medication is subject to, inter alia, the patien metabolizing the medication. It has been determined that select polymorphisms of a patient, including single nucleotide permutations, haplotypes and phenotypes may be utilized to generate such genotype information. The genotype information may be utilized to generate prognostic information. The prognostic information may be utilized in determing treatment options for the patient. The prognostic information is then based on the patient's genetic predisposition to treatment based on their drug metabolism for individual drugs or drug combinations. The prognostic information may also be utilized in determining an expected outcome of a treatment of an individual, such as a treatment with the medication.

[0052] When a genetic marker such as a polymorphism is used as a basis for determining a treatment for a patient, as described herein, the genetic marker may be measured before or during treatment. The prognostic information obtained may be used by a clinician in assessing any of the following: (a) a probable or likely suitability of an individual to initially receive a drug treatment(s); (b) a probable or likely unsuitability of an individual to initially receive a drug treatment(s); (c) a responsiveness of an individual to a drug treatment; (d) a probable or likely suitability of an individual to continue to receive a drug treatment(s); (e) a probable or likely unsuitability of an individual to continue to receive a drug treatment(s); (f) adjusting dosage of an individual receiving a drug; and (g) predicting likelihood of clinical benefits of an individual receiving a drug. As understood by one of skill in the art, measurement of a genetic marker or polymorphism in a clinical setting can be an indication that this parameter may be used as a basis for initiating, continuing, adjusting and/or ceasing administration of a drug, such as described herein.

[0053] Select polymorphisms have been indentified that may be utilized for providing prognostic information, according to the principles of the invention. These findings were correlated with various magnitudes of a positive or negative predispositions to drug metabolism risk. Accordingly, assaying the genotype at these markers may be utilized to generate prognostic information that may be utilized to predict the expected outcome of treating the patient with a drug based on the expected predisposition of the patient to drug metabolism risk associated with the drug. Clinicians prescribing medications may utilize the prognostic information to improve therapeutic decisions and to avoid treatment failures.

[0054] Many of the known human single nucleotide permutations (S Ps) are catalogued by the National Center for Biotechnology Information (NCBI) in the Reference SNP (i.e.,"refSNP") database maintained by NCBI. The Reference SNP database is a polymorphism database (dbSNP), which includes single nucleotide polymorphisms and related polymorphisms, such as deletions and insertions of one or more nucleotides. The database is a public-domain archive maintained by NCBI for a broad collection of simple genetic polymorphisms and can be accessed at http://www.ncbi.nlm.nih.gov/snp.

[0055] A number of patients have experienced health problems associted with the lack of efficacy of certain drugs in specific individuals. Numerous investigations have demonstrated that this phenomenon may be, in part, attributed to the broad variability in individual response profiles and to genetic polymorphisms in candidate genes involved in immunological and inflammatory signaling pathways. Using these polymorphisms to identify patients at risk of irregular drug metabolism would play an important role in modulating drug therapies.

[0056] DNA polymorphisms have been identified that may be utilized according to the principles of the invention include S Ps and haplotypes associated with genetic markers in several genes. The genes include the respective genes encoding cytochrome P450 family 2 subfamily C member 8 (CYP2C8), cytochrome P450 family 2 subfamily C member 9 (CYP2C9), cytochrome P450 family 2 subfamily C member 19 (CYP2C19), cytochrome P450 family 3 subfamily A member 4 (CYP3A4), cytochrome P450 family 3 subfamily A member 5 (CYP3A5), cytochrome P450 family 2 subfamily D member 6 (CYP2D6), cytochrome P450 family 1 subfamily A member 2 (CYP1A2), vitamin K epoxide reductase complex subunit 1 (VKORC1), UDP glucuronosyltransferase family 2 member B7 (UGT2B7), cytochrome P450 family 2 subfamily B member 6 (CYP2B6) and cytochrome P450 family 2 subfamily E member 1 (CYP2E1).

[0057] For example, a method provided by the invention is a diagnostic method for determining the drug metabolism risk associated with a patient which method is not practised on the patient's body, i.e. is an ex vivo diagnostic method. The method may involve determining patient information which may be obtained by assaying a sample of genetic material associated with the patient. The method does not involve obtaining the sample from the patient's body. The invention also provides uses of the systems and methods, for example of the diagnostic assays, for determining the drug metabolism risk associated with a patient.

[0058] Drug Metabolism Algorithm

[0059] The drug metabolism tests evaluate a panel of genetic markers to predict good or poor responders to medications. The tests is directed to patients who are taking or candidates for taking medications metabolized by cytochrome P-450 enzymes, UDP-Glucuronosyltransferase- 2B7 (UGT2B7), and vitamin K epoxide reductase complex subunit 1 (VKORCl) In order to avoid adverse drug events, physicians may also use this test to assist with prescribing medications at optimal doses. Specific drugs are described below.

[0060] For CYP haplotypes, with respect to drug metabolism risk assessment, an exemplary algorithm for determining drug metabolism side effect risk is shown below. Each category is scored separately as shown in the charts below, but all are based on the following scoring system. As would be known by one of ordinary skill in the art, there are four general categories of CYP star alleles (i.e., CYP haplotypes): normal function, reduced function, null function and increased function. The nomenclature is reported by, for example, Robarge et al., "The Star- Allele Nomenclature: Retooling for Translational Genomics" Nature, v. 82, no. 3, September 2007, pp. 244-248, incorporated by reference herein.

[0061] A large number of star alleles have been reported for each cytochrome. Among these are normal functioning CYP star alleles, CYP star alleles with some function that is a reduced function, CYP star alleles with null (or non-functional) alleles, and CYP star alleles with increased functionality. These alleles convey a wide range of enzyme activity, from no activity to ultrarapid metabolism of substrates/medications.

[0062] Table 1 below describes individual SNPs and haplotypes associated with each gene. Once the haplotypes are identified in each gene, the variants are scored and graded to deduce which activity levels and grades are associated with a patient. The graded variants are then used to identify drug and dosage reccomendfations for a patient. The subtables under table 1 below provides allele and haplotype identification, scoring and grading for CYP2C8, CYP2C9,

and CYP2E1, respectively.

[0063] Table 1 A - CYP2C8 alleles, haplotypes, scoring and grading@

@ Unless otherwise indicated, context sequences in FASTA format, are presented by NCBI within the rs cluster report identified by "rs#" associated with each rs number in Table 1A above in the NCBI SNP reference database accessible at http://www.ncbi.nlm.nih.gov/snp, and which is incorporated by reference herein for each recited SNP rs number in the Table(s) above. [0064] Table IB - CYP2C9 alleles, haplotypes, scoring and grading@

@ Unless otherwise indicated, context sequences in FASTA format, are presented by NCBI within the rs cluster report identified by "rs#" associated with each rs number in Table IB above in the NCBI SNP reference database accessible at http://www.ncbi.nlm.nih.gov/snp, and which is incorporated by reference herein for each recited SNP rs number in the Table(s) above [0065] Table 1C - CYP2C19 alleles, haplotypes, scoring and grading@

@ Unless otherwise indicated, context sequences in FASTA format, are presented by NCBI within the rs cluster report identified by "rs#" associated with each rs number in Table 1C above in the NCBI SNP reference database accessible at http://www.ncbi.nlm.nih.gov/snp, and which is incorporated by reference herein for each recited SNP rs number in the Table(s) above

[0066] Table ID - CYP3 A4 alleles, haplotypes, scoring and grading@

@ Unless otherwise indicated, context sequences in FASTA format, are presented by NCBI within the rs cluster report identified by "rs#" associated with each rs number in Table ID above in the NCBI SNP reference database accessible at http://www.ncbi.nlm.nih.gov/snp, and which is incorporated by reference herein for each recited SNP rs number in the Table(s) above

[0067] Table IE - CYP3 A5 alleles, haplotypes, scoring and grading@

@ Unless otherwise indicated, context sequences in FASTA format, are presented by NCBI within the rs cluster report identified by "rs#" associated with each rs number in Table IE above in the NCBI SNP reference database accessible at http://www.ncbi.nlm.nih.gov/snp, and which is incorporated by reference herein for each recited SNP rs number in the Table(s) above

@ Unless otherwise indicated, context sequences in FASTA format, are presented by NCBI within the rs cluster report identified by "rs#" associated with each rs number in Table IF above in the NCBI SNP reference database accessible at http://www.ncbi.nlm.nih.gov/snp, and which is incorporated by reference herein for each recited SNP rs number in the Table(s) above 69] Table 1G - CYPl A2 alleles, haplotypes, scoring and grading@

@ Unless otherwise indicated, context sequences in FASTA format, are presented by NCBI within the rs cluster report identified by "rs#" associated with each rs number in Table 1G above in the NCBI SNP reference database accessible at http://www.ncbi.nlm.nih.gov/snp, and which is incorporated by reference herein for each recited SNP rs number in the Table(s) above [0070] Table 1H - VKORC1 alleles, haplotypes, scoring and grading@

@ Unless otherwise indicated, context sequences in FASTA format, are presented by NCBI within the rs cluster report identified by "rs#" associated with each rs number in Table 1H above in the NCBI SNP reference database accessible at http://www.ncbi.nlm.nih.gov/snp, and which is incorporated by reference herein for each recited SNP rs number in the Table(s) above

[0071] Table II - UGT2B7 and UGT2B 15 alleles, haplotypes, scoring and grading®

[0072] Table 1 J - CYP2B6 alleles, haplotypes, scoring and grading@

CYP2B6

@ Unless otherwise indicated, context sequences in FASTA format, are presented by NCBI within the rs cluster report identified by "rs#" associated with each rs number in Table II and IJ above in the NCBI SNP reference database accessible at http://www.ncbi.nlm.nih.gov/snp, and which is incorporated by reference herein for each recited SNP rs number in the Table(s) above. [0073] Table IK - CYP2E1 alleles, haplotypes, scoring and grading(i

@ Unless otherwise indicated, context sequences in FASTA format, are presented by NCBI within the rs cluster report identified by "rs#" associated with each rs number in Table IK above in the NCBI SNP reference database accessible at http://www.ncbi.nlm.nih.gov/snp, and which is incorporated by reference herein for each recited SNP rs number in the Table(s) above [0074] Once the grades are calculated for each cytochrome, the next step is use these results to make a dosing recommendation for each medication. If a medication is only metabolized by 1 cytochrome, this is fairly straightforward. For the most part, a decreased metabolism result will report that the patient is at risk of an adverse drug event for active medications, or at risk of experiencing insufficient relief from their medication for prodrugs. However, if a medication is metabolized by multiple CYP pathways, the recommendation takes into account all the (clinically-significant) pathways. This is determined using a logical approach for each medication and/or type of medication to provide drug metabolism recommendations.

[0075] Table 2 below provides drug metabolism recommendations for opioids, nonsteroidal anti-inflammatory drugs, benzodiazepines, psychiatric medications, cardiology medications and warfarin.

[0076] Table 2A- Opioid drug recommendations based on CYP grade

[0081] Detection of point mutations or other types of the allelic variants disclosed herein, can be accomplished several ways known in the art, such as by molecular cloning of the specified allele and subsequent sequencing of that allele using techniques known in the art. Alternatively, the gene sequences can be amplified directly from a genomic DNA preparation from the DNA sample using PCR, and the sequence composition is determined from the amplified product. As described more fully below, numerous methods are available for analyzing a subject's DNA for mutations at a given genetic locus such as the gene of interest.

[0082] One such detection method is allele specific hybridization using probes overlapping the polymorphic region and having, for example, about 5, or alternatively 10, or alternatively 20, or alternatively 25, or alternatively 30 nucleotides around the polymorphic region. In another embodiment, several probes capable of hybridizing specifically to the allelic variant are attached to a solid phase support, e.g., a "chip". Oligonucleotides can be bound to a solid support by a variety of processes, including lithography. For example a chip can hold up to 250,000 oligonucleotides (GeneChip, Affymetrix). Mutation detection analysis using these chips comprising oligonucleotides, also termed "DNA probe arrays" is described, e.g., in Cronin et al. (1996) Human Mutation 7:244.

[0083] Alternatively, allele specific amplification technology which depends on selective

PCR amplification may be used in conjunction with the instant invention. Oligonucleotides used as primers for specific amplification may carry the allelic variant of interest in the center of the molecule (so that amplification depends on differential hybridization) (Gibbs et al. (1989) Nucleic Acids Res. 17:2437-2448) or at the extreme 3' end of one primer where, under appropriate conditions, mismatch can prevent, or reduce polymerase extension (Prossner (1993) Tibtech 11 :238 and Newton et al. (1989) Nucl. Acids Res. 17:2503). This technique is also termed "PROBE" for Probe Oligo Base Extension. In addition it may be desirable to introduce a novel restriction site in the region of the mutation to create cleavage-based detection (Gasparini et al. (1992) Mol. Cell. Probes 6: 1).

[0084] If the polymorphic region is located in the coding region of the gene of interest, yet other methods than those described above can be used for determining the identity of the allelic variant according to methods known in the art.

[0085] The genotype information obtained from analyzing a sample of a patient's genetic material may be utilized, according to the principles of the invention, to predict whether a patient has a level of risk associated with poor opioid maintenance response. The risk may be associated with a side effect the patient may be susceptible to developing, an efficacy of the drug to the patient specifically or some combination thereof. The genotype information of the patient may be combined with demographic information about the patient as described above.

[0086] Referring to FIG. 1, depicted is an assay system 100. An assay system, such as assay system 100, may access or receive a genetic material, such as genetic material 102. The sample of genetic material 102 can be obtained from a patient by any suitable manner. The sample may be isolated from a source of a patient's DNA, such as saliva, buccal cells, hair roots, blood, cord blood, amniotic fluid, interstitial fluid, peritoneal fluid, chorionic villus, semen, or other suitable cell or tissue sample. Methods for isolating genomic DNA from various sources are well-known in the art. Also contemplated are non-invasive methods for obtaining and analyzing a sample of genetic material while still in situ within the patient's body.

[0087] The genetic material 102 may be received through a sample interface, such as sample interface 104 and detected using a detector, such as detector 106. A polymorphism may be detected in the sample by any suitable manner known in the art. For example, the polymorphism can be detected by techniques, such as allele specific hybridization, allele specific oligonucleotide ligation, primer extension, minisequencing, mass spectroscopy, heteroduplex analysis, single strand conformational polymorphism (SSCP), denaturing gradient gel electrophoresis (DGGE), oligonucleotide microarray analysis, temperature gradient gel electrophoresis (TGGE), and combinations thereof to produce an assay result. The assay result may be processed through a data management module, such as data management module 108, to produce genotype information 112. The genotype information 112 may include an assay result on whether the patients has a genotype including one or more of the allelic variants listed in Tables I and 3 above. The genotype information 112 may be stored in data storage 110 or transmitted to another system or entity via a system interface 114.

[0088] Referring to FIG. 2, depicted is a prognostic information system 200. The prognostic information system 200 may be remotely located away from the assay system 100 or operatively connected with it in an integrated system. The prognostic information system 200 receives the genotype information 112 through a receiving interface 202 for processing at a data management module 204 to generate prognostic information 210. The data management module 204 may utilize one or more algorithms described in greater detail below to generate prognostic information 210. The prognostic information 210 may be stored in data storage 208 or transmitted via a transmitting interface 206 to another system or entity. The transmitting interface 206 may be the same or different as the receiving interface 202. Furthermore, the system 200 may receive prognostic information 220 prepared by another system or entity. Prognostic information may be utilized, in addition to or in the alternative, to genotype information 112 in generating prognostic information 210. [0089] Referring to FIG. 3, depicted is a prognostic information process 300 which may be utilized for preparing information, such as genotype information 112 and prognostic information 210, utilizing an assay system, such as assay system 100 and/or a prognostic information system, such as prognostic information system 200, according to an embodiment. The steps of process 300, and other methods described herein, are described by way of example with the assay system 100 and the prognostic information system 200. The process 300 may be performed with other systems as well.

[0090] After process start, at step 302, a sample of genetic material of a patient is obtained as it is received at the sample interface 106. The sample interface can be any type of receptacle for holding or isolating the genetic material 102 for assay testing.

[0091] At step 304, the genetic material 102 is tested utilizing the detector 106 in assay system 100 to generate genotype information 112. The detector 106 may employ any of the assay methodologies described above to identify allelic variants in the genetic material 102 and generate the genotype information 112 including polymorphism data associated with one or more of the DNA polymorphisms described above in Tables 1 and 3. The data management module 108, utilizing a processor in an associated platform such as described below, may store the genotype information 112 on the data storage 110 and/or transmit the genotype information 112 to another entity or system, such as prognostic information system 200 where it is received at receiving interface 202 for analysis.

[0092] At step 306, the genotype information 112 can be analyzed utilizing a processor in an associated platform, such as described below, by using an algorithm which may be programmed for processing through data management module 204. The algorithm may utilize a scoring function to generate predictive values based on the polymorphism data in the genotype information 112. Different algorithms may be utilized to assign predictive values and aggregate values.

[0093] For example, an additive effect algorithm may be utilized to generate an analysis of a patient's genetic predisposition and their demographic phenotype predisposition to drug metabolism risk. In the additive effect algorithm, polymorphism data of the genotype information obtained from analyzing a patient's genetic material is utilized to indicate the active polymorphisms identified from a patient's genotype information. A tested polymorphism may be determined to be (1) absent or present in either (2) a heterozygous or (3) a homozygous variant in the patient's genotype. According to the additive effect algorithm, the polymorphisms identified from a patient's genotype information and demographic phenotype are each assigned a real value, such as an Odds Ratio (OR) or a parameter score, depending on which polymorphisms appears in the patient's genotype and demographic information.

[0094] To gather data for the algorithm, one or more of the alleles and haplotypes such as those listed in Table 1 may be tested and/or analyzed to produce one or more values associated with the presence or absence of the alleles and haplotypes. Other factors, such as other S P Diploid Polymorphisms, other demographic phenotypes may also be tested and/or analyzed to produce one or more values associated with the presence or absence of the other SNP Diploid Polymorphisms and other demographic phenotypes.

[0095] The values gathered are based on results of the various tests and data gathered and/or determined. The values ma drug metabolism response based on the subject's genetic information and/or non-genetic characteristics or phenotypes. The algorithm may compute a composite score based on the results of individual tests. The composite score may be calculated based on an additive analysis of the individual scores which may be compared with a threshold value for determining drug metabolism response based on the additive score. In addition or in the alternative, more complex functions may be utilized to process the values developed from the testing results, such as utilizing one or more weighting factor(s) applied to one or more of the individual values based on various circumstances, such as if a subject was tested using specific equipment, a temporal condition, etc.

[0096] In all of the preceding examples, the predictive values and aggregate values generated are forms of prognostic information 210.

[0097] At step 310, the result of the comparison obtained in step 308 generates a second form of prognostic information 220. For example, (a) if the determined sum is higher than the threshold value, it can be predicted that the patient is at an elevated risk for drug metabolism risk associated with prescribing the patient a medication; (b) if the determined sum is at or near the threshold value, it can be predicted that the patient is at a moderate risk for drug metabolism risk; and (c) if the determined sum is below the threshold value, it can be predicted that the patient is at a low risk for drug metabolism risk.

[0098] Also at step 310, the data management module 205 in the prognostic information system 200 identifies a risk to a patient by executing an algorithm, such as the additive effect algorithm described above, and communicating the generated prognostic information 210. The data management module 204, utilizing a processor in an associated platform such as described below, may store the prognostic information 210 on the data storage 208 and/or transmit the prognostic information 210 to another entity or system prior to end of the prognostic information process 300. Other algorithms may also be used in a similar manner to generate useful forms of prognostic information for determining treatment options for a patient. [0099] Referring to FIG. 4, there is shown a platform 400, which may be utilized as a computing device in a prognostic information system, such as prognostic information system 200, or an assay system, such as assay system 100. It is understood that the depiction of the platform 400 is a generalized illustration and that the platform 400 may include additional components and that some of the components described may be removed and/or modified without departing from a scope of the platform 400.

[00100] The platform 400 includes processor(s) 402, such as a central processing unit; a display 404, such as a monitor; an interface 406, such as a simple input interface and/or a network interface to a Local Area Network (LAN), a wireless 802.1 lx LAN, a 3G or 4G mobile WAN or a WiMax WAN; and a computer-readable medium (CRM) 408. Each of these components may be operatively coupled to a bus 416. For example, the bus 416 may be an EISA, a PCI, a USB, a FireWire, a NuBus, or a PDS.

[00101] A CRM, such as CRM 408 may be any suitable medium which participates in providing instructions to the processor(s) 402 for execution. For example, the CRM 408 may be non-volatile media, such as an optical or a magnetic disk; volatile media, such as memory; and transmission media, such as coaxial cables, copper wire, and fiber optics. Transmission media can also take the form of acoustic, light, or radio frequency waves. The CRM 408 may also store other instructions or instruction sets, including word processors, browsers, email, instant messaging, media players, and telephony code.

[00102] The CRM 408 may also store an operating system 410, such as MAC OS, MS

WINDOWS, UNIX, or LINUX; application(s) 412, such as network applications, word processors, spreadsheet applications, browsers, email, instant messaging, media players such as ga .g., "apps"); and a data structure managing application 414. The operating system 410 may be multi-user, multiprocessing, multitasking, multithreading, real-time and the like. The operating system 410 may also perform basic tasks such as recognizing input from the interface 406, including from input devices, such as a keyboard or a keypad; sending output to the display 404 and keeping track of files and directories on CRM 408; controlling peripheral devices, such as disk drives, printers, image capture devices; and for managing traffic on the bus 416. The applications 412 may include various components for establishing and maintaining network connections, such as code or instructions for implementing communication protocols including those such as TCP/IP, HTTP, Ethernet, USB, and Fire Wire.

[00103] A data structure managing application, such as data structure managing application 414 provides various code components for building/updating a computer-readable system architecture, such as for a non-volatile memory, as described above. In certain examples, some or all of the processes performed by the data structure managing application 412 may be integrated into the operating system 410. In certain examples, the processes may be at least partially implemented in digital electronic circuitry, in computer hardware, firmware, code, instruction sets, or any combination thereof.

[00104] Although described specifically throughout the entirety of the disclosure, the representative examples have utility over a wide range of applications, and the above discussion is not intended and should not be construed to be limiting. The terms, descriptions and figures used herein are set forth by way of illustration only and are not meant as limitations. Those skilled in the art recognize that many variations are possible within the spirit and scope of the principles of the invention. While the examples have been described with reference to the figures, those skilled in the art are able to make various modifications to the described examples without departing from the scope of the following claims, and their equivalents.